A molecular docking calculation of the title compound with the neuraminidase enzyme was carried out.In the title chalcone-thiophene derivative, the dihedral angle between the aromatic and the thiophene rings is 11.4 (2)°. In the crystal, molecules are linked by C—H⋯O and C—H⋯S weak interactions along [100], forming rings of  15H15NOS, whose molecular structure matches the asymmetric unit. The molecule is not planar, the dihedral angle between the aromatic and the thiophene rings being 11.4 (2)°. In the crystal, molecules are linked by C—H⋯O and weak C—H⋯S interactions along [100], forming 22R(8) rings, and by weak C—H⋯O interactions along [010], forming chains with a C(6) graph-set motif. In addition, molecules are connected into centrosymmetric dimers by weak C—H⋯π interactions, as indicated by the Hirshfeld surface analysis. The most important contributions for the crystal structure are the H⋯H (46.50%) and H⋯C (23.40%) interactions. The crystal packing resembles a herringbone arrangement when viewed along [100]. A molecular docking calculation of the title compound with the neuraminidase enzyme was carried out. The enzyme shows (ASN263)N—H⋯O, (PRO245)C—H⋯Cg(thiophene ring) and (AGR287)C—H⋯N intermolecular interactions with the title compound. The crystal structure was refined as a two-component twin with a fractional contribution to the minor domain of 0.0181 (8).The equimolar reaction between 4-(dimethylamino)benzaldehyde and 2-acetylthiophene in basic ethanolic solution yields the title compound, C The 22R(8) rings are the subunits of the periodic arrangement along [100] and one very weak H7⋯H2i contact is also observed [H⋯H = 2.26 Å]. The molecular units are also linked by very weak C15—H15⋯O1ii links into chains along [010] with a C(6) graph-set motif  and di (x axis) values are the closest external and internal distances  from given points on the Hirshfeld surface contacts N—H⋯O1 (d = 1.796), (PRO245)C—H⋯Cg(thiophene ring) (d = 2.829) and (AGR287)C—H⋯N1 (d = 2.620)  contact is observed in the structure interpretation, by the centrosymmetric dimeric arrangement of the molecules  Fig. 8. More des Figs. 3 and 9 ▸,sp2-hybridized C atoms of the main fragment, and the weak intermolecular interactions, e.g. H⋯H, H⋯C or π–π contacts. One example for comparison with the title compound is the crystal structure of the compound 3-(4-methylphenyl)-1-(3-thienyl)-2-propen-1-one  and for the above-mentioned 3-thienyl derivative, along [001] .Chalcone-thiophene derivatives have some molecular structural features in common, namely the nearly planar geometry, as a result of the 1] Fig. 9b.et al., 2016All starting materials are commercially available and were used without further purification. The synthesis of the title compound was adapted from a previously reported procedure  = 1.2Ueq(C) or 1.5Ueq(C) for methyl H atoms. A rotating model was used for the methyl groups. The crystal was refined as a two-component twin {twin law: two-axis (001) [105], BASF = 0.0181 (8)}.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989017003437/rz5205sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017003437/rz5205Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017003437/rz5205sup3.pdfSUPPORTING INFORMATION FOR THE EVALUATION OF THE CRYSTAL STRUCTURE OF (2E)-3-[4-(DIMETHYLAMINO)PHENYL]-1-(THIOPHEN-2-YL)PROP-2-EN-1-ONE AND THE NEURAMINIDASE ENZYME. DOI: Click here for additional data file.10.1107/S2056989017003437/rz5205Isup4.cmlSupporting information file. DOI: 1535563CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


In the crystal structure of the title compound, a dispiro-2,5′′-dione, C—H⋯O hydrogen bonding predominates, linking molecules to form chains propagating along [100]. 32H26ClN3O2, the cyclohexanone ring of the isoquinoline unit has a distorted envelope conformation, with the methylene C atom adjacent to the spiro C atom as the flap. The central 1-methylpyrrolidine ring has an envelope conformation with the N atom as the flap. The mean planes of the indolin-2-one ring system, the chlorobenzene ring and the isoquinoline ring system are inclined to the mean plane of the central 1-methylpyrrolidine ring by 87.95 (11), 71.01 (12) and 88.81 (10)°, respectively. There are two short C—H⋯O intramolecular contacts present. In the crystal, molecules are linked via C—H⋯ O hydrogen bonds, forming chains along the a-axis direction. The NH H atom is involved in a weak N—H⋯O hydrogen bond with the same carbonyl O atom. There are no further significant intermolecular contacts present. The largest contribution to the overall Hirshfeld surface of 52.3% is due to H—H contacts.In the title dispiro compound, C The pyridine ring (N3/C32–C36) has a shallow twist-boat conformation . Their mean planes are inclined to each other by 14.06 (10)°, and the phenyl ring (C51–C56) is inclined to the pyridine ring mean plane by 22.35 (12)°.The molecular structure of the title molecule is shown in Fig. 1Q = 0.094 (2) Å, θ = 92.3 (13)°, φ = 84.5 (13)°]. The mean planes of the indolin-2-one ring system, the chlorobenzene (C41–C46) ring and the isoquinoline (N3/C3/C31–C38) ring system are inclined to the mean plane of the central 1-methylpyrrolidine (N1/C2–C5) ring by 87.95 (11), 71.01 (12) and 88.81 (10)°, respectively. The sum of the bond angles around atoms N1 and N2 are 333.6 and 358.6°, respectively, indicating a pyramidal geometry and sp3 hybridization.In the indolin-2-one ring system (N2/C2/C21–C27), the benzene (C21–C26) and pyrrolidine (N2/C2/C21/C26/C27) rings make a dihedral angle of 2.45 (12)°, while the keto atom O1 deviates from the attached pyrrolidine ring by 0.043 (1) Å. The 1-methylpyrrole ring (N1/C2–C5) has an envelope conformation with atom N1 as the flap -1,2′′(1′′H)-dione methanol solvate -dione chloroform solvate . The relative contributions of the other different intermolecular interactions to the Hirshfeld surface in descending order are: C⋯H/H⋯C (23.3%), O⋯H/H⋯O (8.5%), Cl⋯H/H⋯Cl (8.4%), N⋯H/H⋯N (4.1%) and there is only a very small contribution from other contacts, i.e. 3.1%, in the structure. This illustrates that the N—H⋯O and C—H⋯O interactions contribute significantly to the crystal packing of the title compound.The overall two-dimensional fingerprint plot is illustrated in Fig. 5E)-6-(2-chlorobenzylidene)-2-phenyl-7,8-dihydroquinolin-5(6H)-one (L) in 94% yield (m.p. 323–324 K). A mixture of isatin (1.1 mmol) and sarcosine (1.1 mmol) was taken in 10 ml of acetonitrile in a 50 ml round-bottom flask and heated to reflux for 2 h. Then 1 mmol of L was added to the above reaction mixture and reflux was continued for a further 14 h. After completion of the reaction, as evident from TLC, the solvent was removed under reduced pressure and the residue washed with ice-cold water (50 ml). The crude product was purified by column chromatography using a 90:10 (v/v) petroleum ether–ethyl acetate mixture to obtain the pure product . Colourless block-like crystals were obtained by slow evaporation of a solution in ethyl acetate.An equimolar mixture of 2-phenyl-5,6,7,8-tetrahydro-5-quinolinone and 2-chlorobenzaldehyde was dissolved in 10 ml of ethanol followed by the addition of 0.5 equiv. of potassium hydroxide. The mixture was stirred for 1 h at ambient temperature and the precipitate formed was filtered and dried to obtain pure (Uiso = 1.5Ueq(C-methyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989018005455/su5434sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018005455/su5434Isup2.hklStructure factors: contains datablock(s) I. DOI: 1835595CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


For example, conventional αβ T cells develop from DP cells after partial-agonist T cell receptor (TCR) interactions with self-peptide/MHC, whereas unconventional αβ T cells, such as TCRαβ(+)CD8αα(+) intraepithelial lymphocytes (IELs), require full-agonist TCR interactions. Despite this, DP cells appear homogeneous, and it remains unclear how distinct TCR signalling instructs distinct developmental outcomes. Moreover, whether TCR signals at earlier stages of development, for example in CD4(−)CD8(−) double negative (DN) cells, impact on later fate decisions is presently unknown. Here, we assess four strains of mice that display altered TCR signal strength in DN cells, which correlates with altered generation of unconventional TCRαβ(+)CD8αα(+) IELs. FVB/n mice  and mice with altered preTCRα (pTα) expression, both displayed weaker TCR signalling in DN cells, an inefficient DN-to-DP transition, and reduced contribution of TCRαβ(+)CD8αα(+) IELs to gut epithelium. Conversely, TCRαβ(+)CD8αα(+) IEL development was favoured in mice with increased TCR signal strength in DN cells. Collectively, these data suggest TCR signal strength in DN cells directly impacts on subsequent DP cell differentiation, fundamentally altering the potential of thymocyte progenitors to adopt conventional versus unconventional T cell fates.CD4 Gut IELs display anti-microbial and anti-inflammatory properties and are central to the control of intestinal epithelial homeostasis2. A large proportion of gut IELs express TCRαβ, and can be further characterized by expression of CD8αβ or CD8αα dimers3. TCRαβ(+)CD8αβ(+) IELs are termed “conventional”, closely sharing gene expression signatures with CD8αβ(+) T cells from secondary lymphoid organs5. By contrast, “unconventional” TCRαβ(+)CD8αα(+) IELs do not require priming in lymphoid structures, appear restricted to the gut epithelium, and display gene expression signatures more similar to γδ T cells5.T cell receptor-expressing intraepithelial lymphocytes (IELs) are epithelial-resident T cells found at numerous body locations(+)CD8αα(+) IELs develop extra-thymically in gut lymphoid structures known as cryptopatches6. However, TCRαβ(+)CD8αα(+) IELs are severely reduced in athymic nude mice, and fate-mapping experiments suggested they traverse the CD4(+)CD8(+) double positive (DP) stage in the thymus7. Further work implicated agonist self-peptide-mediated selection through TCRαβ at the DP stage8, and identified pre- and post-selection progenitor subsets9. Nonetheless, it remains unclear how “strong” TCR-agonist signals in DP cells instruct positive selection of TCRαβ(+)CD8αα(+) IELs instead of driving negative selection. Indeed, unconventional TCRαβ(+)CD8αα(+) IELs were recently found to express TCRs that had been “recycled” from strong negatively selecting signals10.Initial studies suggested that unconventional TCRαβ(+)CD8αα(+) IEL development, the DP stage is not the first in which TCR signalling occurs. DP cells arise from CD4(−)CD8(−) double negative (DN) cells in a process known as “β-selection” that is mediated by signalling through the preTCR (rearranged TCRβ paired with invariant pTα)11. PreTCR signalling is generally considered weak, due to very low surface preTCR expression12. By contrast, stronger signalling in DN cells, for example by TCRγδ, is less efficient at generating DP cells; instead driving cells to a γδ T cell fate14.Although TCRαβ signalling at the DP stage appears critical for TCRαβ15. Although TCR signal strength in DN cells clearly affects the efficient induction of these processes, it is presently unclear whether it also affects the future fate of the DP cells that are generated. Here, we begin to investigate this idea in the context of TCRαβ(+)CD8αα(+) IEL development. We show that FVB/n wild type (WT) mice have a much reduced TCRαβ(+)CD8αα(+) IEL compartment when compared with WT C57BL/6 animals, that correlates with weaker preTCR signalling at the β-selection checkpoint. Indeed, by reducing preTCR signal strength in pTα-transgenic animals we re-capitulate this relative absence of gut TCRαβ(+)CD8αα(+) IELs. By contrast, in two mouse models in which TCR signal strength is greater in DN cells by forced expression of TCRαβ, increased generation of TCRαβ(+)CD8αα(+) IELs was observed. Thus, these data provide evidence that TCR signal strength at the DN-to-DP transition directly influences the efficiency of TCRαβ(+)CD8αα(+) IEL development.Successful transition through the β-selection checkpoint results in cell survival, extensive proliferation, and significant differentiation, events that may be mechanistically linked(+)CD8αα(+) IELs in the small intestine CD8αβ(+) IELs was increased in FVB/n mice, and TCRγδ(+) IELs were comparable between the two strains CD8αα(+) IELs are thought to develop in the thymus from cells that are CD4(lo)CD8(lo)TCRδ(−)TCRβ(+)CD5(hi)CD69(+)PD-1(+)CD122(+) CD8(+) double positive (DP) cells16, that in turn are generated when the preTCR (rearranged TCRβ paired with invariant pre-Tα) drives CD44(−)CD25(+) DN3 cells to CD44(−)CD25(−) DN4 cells (and then on to the DP stage) in a process called β-selection11. C57BL/6 mice had an expected DN3-to-DN4 ratio of ~1.5, consistent with a normal transition through the β-selection checkpoint . Moreover, DN3 cells from FVB/n mice had significantly elevated surface expression of CD25, a signatory feature of inefficient progression through the DN-to-DP transition11 CD8αα(+) IEL compartment compared with C57BL/6 animals, also display inefficient developmental progression through thymic stages that lead to generation of unconventional IEL progenitors.We had noted that FVB/n wild type (WT) mice, in comparison with C57BL/6 WT mice, had significantly reduced unconventional TCRαβine Fig.  . By conins Fig. . Unconvells Fig. . IEL proint Fig. . Howevern11 Fig. . Thus, F(+)CD8αα(+) IEL development, we first assessed components of the preTCR. Intracellular TCRβ levels in DN3 cells appeared comparable between FVB/n and C57BL/6 animals , than C57BL/6 mice  of CD71(+)i.c.TCRβ(+) proliferating DN3 cells18 CD8αα(+) IELs in FVB/n mice.To investigate the inefficient DN3-to-DN4 transition in FVB/n mice that appeared to correlate with reduced unconventional TCRαβals Fig. . For pTαice Fig. . It was s18 Fig. , that exs18 Fig. , a recog(+)CD8αα(+) IELs are linked, we generated mice in which only a weakly signalling pTαa-containing preTCR could be formed. pTα-intron-1 was removed from the full pTα genomic locus (to prevent splicing to pTαb), in a 50 kb bacterial artificial chromosome (BAC), that was then used to generate pTαa-only BAC transgenic mice on a pTα−/− (C57BL/6) background  DN3 cells displayed markedly less CD5, indicative of weaker TCR signalling CD8αα(+) IELs in the small intestine CD8αβ(+) IELs and TCRγδ(+) IELs were present in normal proportions CD8αα(+) IELs.To test the idea that weaker signalling at the β-selection checkpoint and reduced generation of unconventional TCRαβls, Fig. . Consistls, Fig. , with a als Fig. . Moreove WT Fig. , while ning Fig. . Importaine Fig.  . By con(+)CD8αα(+) IELs. To test this idea further, we sought to increase TCR signalling in DN cells to ascertain if this would conversely increase unconventional TCRαβ(+)CD8αα(+) IEL development. TCRαβ transgenic (tg) mice aberrantly express TCRαβ complexes that signal strongly at the β-selection checkpoint. Indeed, as well as driving DN cells to the DP stage, this strong TCR signalling is also thought to divert some DN progenitors to the γδ lineage22. OT-II mice (C57BL/6 background), express an ovalbumin-specific MHC-II-restricted Vα2(+)Vβ5(+) transgenic TCRαβ23. Vα2(+) (i.e. largely tg-TCR(+)) DN3 cells from OT-II mice express much higher levels of CD5 than Vα2(−) DN3 cells, indicative of stronger TCR signalling  DN cells from OT-II mice traverse the β-selection checkpoint efficiently CD8αα(+) IELs was observed, notably inverting the TCRαβ(+)CD8αα(+) IEL-to-TCRαβ(+)CD8αβ(+) IEL ratio  rearrange early TCRα (and TCRβ). Compared with pTα−/− mice, TCRδ−/−. pTα−/− mice had an increased population of DN4 cells, the majority of which robustly expressed TCRβ CD8αα(+) IELs was observed, again inverting the TCRαβ(+)CD8αα(+) IEL-to-TCRαβ(+)CD8αβ(+) IEL ratio CD8αα(+) IEL development.TCR-tg mice express a single TCRα/TCRβ combination that may have unusual signalling characteristics. Thus, to extend these studies, we took advantage of a little-known feature of TCRδCRβ Fig. . Unlike als Fig. . Moreoveing Fig. . Thus, βtio Fig. . Collect(+)CD8αα(+) intraepithelial lymphocytes have long been debated. Initially, the recognition that TCRαβ(+)CD8αα(+) IELs, but not TCRαβ(+)CD8αβ(+) IELs, were present in athymic mice led to suggestion of their extra-thymic generation in gut lymphoid structures called cryptopatches (CPs)6. Indeed, CPs were shown to contain CD25(+)IL-7Rα(+)c-kit(+) progenitors that could reconstitute the T cell compartments of irradiated T cell-deficient animals26. Nonetheless, that athymic mice have only 5–10% of the total TCRαβ(+)CD8αα(+) IELs of euthymic mice conversely argued for a predominately thymic origin. The thymic generation of TCRαβ(+)CD8αα(+) IELs was further supported by fate mapping experiments in which a GFP marker was activated by RORγt-promoter-driven Cre protein7. RORγt is expressed in thymic DP cells but not in their DN precursors. As all γδ T cells develop from DN cells, TCRγδ(+) IELs were GFP(−). By contrast, TCRαβ(+)CD8αα(+) IELs were GFP(+), suggesting their development through a DP stage7.The developmental origins of TCRαβ(+)CD8αα(+) IELs use “forbidden” TCRβ chains that were purged from the conventional TCRαβ(+) T cell pool by superantigen-driven negative selection of DP cells in the thymus27. Nonetheless, despite evidence of “TCR-agonist-selection”28, identifying the thymic stages through which TCRαβ(+)CD8αα(+) IEL progenitors progress has proven more problematic. Cheroutre and colleagues described a minor subset of CD8αα(+) DP cells that appeared to develop as TCRαβ(+)CD8αα(+) IELs if exposed to TCR-agonist signals9. Somewhat surprisingly, these “triple positive” (TP) cells then shut off CD4, CD8α and CD8β to become TCRαβ(+)CD5(+) DN cells, before re-expressing CD8αα(+) in the gut epithelium possibly under the influence of TGFβ29. By contrast, the stages that precede the TP stage are less clear. As too are the reasons why TP cells survive TCR-agonist signals, rather than dying as DP cells do during TCR-agonist-driven negative selection. It is conceivable that TP cells have already entered an unconventional IEL developmental pathway, and are thus cell-intrinsically distinct from DP cells that allow them to survive agonist signals. In this regard, RhoH30, and TGFβ signalling29, have been implicated in differential thymic development of TCRαβ(+)CD8αα(+) IELs versus conventional T cell subsets. Nonetheless, how unconventional IEL progenitors could adopt such a distinct gene expression profile by the TP stage, and indeed how TP cells are even generated, had not been previously elucidated.It was also recognized early that TCRαβ(+)CD8αα(+) IELs; stronger TCR signalling in DN cells, as judged by CD5 surface levels and the efficiency of the DN-to-DP transition, appearing to favour TCRαβ(+)CD8αα(+) IEL generation. As TP cells, like DP cells, are presumably derived from DN precursors as a result of (pre)TCR signalling, we speculate that stronger signalling at the DN stage favours TP generation. Unfortunately, we were unable to formally test this idea, as CD8αα-specific antibodies are not available and TL-tetramer staining did not work in our hands9. Nonetheless in this regard, peripheral CD4(+) cells do upregulate CD8αα when activated through TCRαβ (by cross-linking antibodies) in the presence of TGFβ29, the absence of which has been correlated with reduced generation of TCRαβ(+)CD5(+) IEL precursors29.In this study, we have shown that modulation of TCR signal strength at the thymic DN stage fundamentally impacts on the development of TCRαβ(+)) DP cells conflicts with the notion that such signals are required for commitment to the γδ T cell lineage14. Indeed, premature expression of TCRα in DN cells of TCRαβ transgenic mice was shown to promote differentiation of CD4(−)CD8(−)TCRαβ(+) cells with γδ T cell-like properties22. Nonetheless, many of these studies also reported increased numbers of TCRαβ(+)CD8αα(+) IELs, and made obvious but perhaps incorrect links between these DN TCRαβ(+) cells and IEL development31. However, these “γδ-wannabie” cells do not traverse the DP stage (unlike bone fide IEL progenitors), and do not rearrange their endogenous TCRα chains20; something obviously required for TCRαβ(+) IEL development. By way of explanation, our data now suggest that in TCRαβ(+) transgenic mice at least some DN cells will transition to the DP stage as a result of stronger transgenic-TCR signalling. This could boost the TP cell to conventional DP cell ratio, which in turn would favour increased TCRαβ(+)CD8αα(+) IEL development.At first glance, invoking strong TCR signals in DN cells to drive generation of CD8αα(+) IELs.Finally, what would normally provide the stronger TCR signals in DN cells to drive TP generation in the absence of prematurely expressed transgenic TCRαβ? TCRα is reported to rearrange in a small fraction of WT DN cells(+)CD8αα(+) IELs. Stronger signalling favours TCRαβ(+)CD8αα(+) IEL development. By contrast, weaker signalling favours a greater contribution from conventional TCRαβ(+)CD8αβ(+) IELs that are likely differentiated from naïve conventional CD8αβ(+) T cells in the gut-associated lymphoid tissue.In sum, our findings shed new light on the early thymic stages of unconventional IEL development. We demonstrate that TCR signal strength in DN cells at the β-selection checkpoint significantly influences the development of unconventional TCRαβAll experimental protocols were performed in, and approved by, The Blizard Institute, Bart’s and The London School of Medicine, Queen Mary University of London.C57BL/6 and FVB/n mice were purchased from Charles River Laboratories. All mice were 6–12 weeks old. Mice were bred and maintained in specific pathogen-free animal facilities at Queen Mary University of London. All experiments were performed in compliance with relevant laws and institutional guidelines and were approved by a local ethics committee.http://recombineering.ncifcrf.gov)33. In brief, a pTα locus-containing BAC (BMQ452P20) was purchased from www.ensemble.org. A GalK cassette was first amplified from a GalK plasmid using primers that contained 50 bp homology with sections of pTα intron-1;BAC-transgenic mice were generated by the recombineering technique  in fluorescence-activated cell sorting (FACS) buffer  containing; 2% heat-inactivated fetal calf serum (FCS) (Life technologies); and 5 mM ethylenediaminetetraacetic acid (EDTA) (Life technologies)). For spleen, red blood cells were removed by gradient centrifiguation at 1600rpm for 25 min using 4 mL of Lymphocyte Separation Medium (Fischer). For IEL preparations, faecal material was flushed from lumen of small intestine with ice-cold PBS using a gavage needle. Fatty tissues, vasculature and Peyer’s patches were removed, followed by longitudinal opening and 60 min agitation in RPMI-1640 10% Newborn calf serum (NCS) (Life technologies) with 5 mM EDTA (Life technologies) at 37 °C. Cells were subsequently passed through an autoclaved column containing 0.7 g of nylon wool , equilibrated with RPMI-1640 with 38 mM HEPES (Life technologies). IELs were enriched on a discontinuous Percoll gradient (40%/80% isotonic Percoll), before use and stained as described below.Fluorochrome-conjugated antibodies (eBioscience or BD) were; CD3ε (145-2C11), CD4 (RM4-5), CD8α (53–6.7), CD25 (PC61), CD44 (IM7), TCRδ (GL3), TCRβ (H57–597), CD8β (H35–17.2), CD5 (53–7.3), CD69 (H1.2F3), Vα2 (B20.1), CD117 (2B8), B220 (RA3–6B2). For surface staining, cells were Fc-blocked  and stained with antibodies in FACS buffer  for 30 mins. Cells were then washed twice with FACS buffer using a centrifugation speed of 300 g for 5 mins. After staining, cells were resuspended in FACS buffer containing 0.5 μg/ml DAPI (Life technologies) for dead cell exclusion. For intracellular staining, cells were first stained for extracellular markers and subsequently fixed and permeabilized using the eBioscience intracellular flow kit according to the manufacturer’s protocol. The corresponding isotype control was used as a negative control. Samples were acquired using an LSR-II and analysed using Flow Jo v10.Total RNA was extracted using the RNeasy Mini kit (QIAGEN) according to the manufacturer’s instructions. Concentration and purity was determined using the NanoDrop ND-1000 spectrophotometer (Thermo Scientific). Total RNA was reverse-transcribed into cDNA using the High-Capacity RNA-to-cDNA™ Kit (life technologies). Quantitative real-time PCR was performed using the power SYBR green PCR Master mix (life technologies) on the 7500 real-time PCR system (life technologies). Primers were designed manually and recognised intron/exon boundaries;aFwd: 5′-GGCTCTACCATCAGGCATCGC-3′;pTαaRev: 5′-GGTGGTTTGCCTGGTCCTCG-3′;pTαbFwd: 5′-GCTCTACCATCAGGGGGAATCTTC-3′;pTαbRev: 5′-CGGGGGGACACAGCGG-3′.pTαAll samples were run in triplicate and expression was normalised against the housekeeping gene GAPDH. Analysis of PCR results was performed on 7500 real-time software v2.0.6 (life technologies).Cells were individually FAC-sorted into 96 well-plates containing 10 μl of reverse transcription mix , 30U MMLV-RT reverse transcriptase (life Technologies), 1% BSA and 0.5 μM of the following primers;pTαgeneF: 5′-TAGGACATGGCTGCTGCTGC-3′;pTαgeneR: 5′-TCCCACCCACAGAATTTGGAC-3′.2 (NEB) and 0.25 mM of the pTαgeneF and pTαgeneR primers. The second round PCR was performed using 2.0 μl of the PCR product generated by the first-round PCR and consisted of a 3 min denaturation step followed by 25 cycles of amplification  in a PCR mix containing 6.5U of Taq polymerase, 1.6 mM MgCl2, 0.25 mM dNTPs, and 0.25 mM of the following primers;Plates were incubated for 2hr at 37 °C in a thermocycler (Biorad) and the reaction was stopped by 10 min incubation at 70 °C. Two rounds of PCR were then carried out on the cDNA. The first round of PCR consisted of denaturation at 94 °C, followed by 34 cycles of amplification  in a PCR mix containing 7.5U of Taq polymerase (NEB), 0.25 mM dNTPs (NEB), 1.6 mM MgClaF: 5′-GCTCTACCATCAGGCATCGC-3′;pTαaR: 5′-CTCCAGCTGTCAGGGGAATC-3′;pTαbF: 5′-GCTCTACCATCAGGGGAATCTTC-3′;pTαbR: 5′-GCAGGTACTGTGGCTGAGC-3′.pTαPCR products were run on a ReadyAgarose 96 Plus gel (Biorad) and PCR products were sequenced for validation.Data are presented as mean ± s.d. A student’s t-test was used to assess statistical significance between groups. A difference was considered significant if p < 0.05.Supplementary Figures


It has been highlighted that the original article , publishChanges:A labeling error in Table [Without DPN n = 607 (100%)] was replaced by (Yes n = 396).[With DPN n = 396 (100%)] was replaced by (No n = 607).[Age group (year): (mean ± SD)] was replaced by [Age (year)], and the mean age was removed from the table.2): (mean ± SD)] was replaced by [Body mass index (kg/m2)] and the mean BMI was removed.[Body mass index (BMI) (kg/m[Duration of diabetes (year): (mean ± SD)] was replaced by [Duration of diabetes (year)] and the mean duration was removed.SBP mmHg (mean ± SD) DBP mmHg (mean ± SD) was removed.All these were removed TC (mean ± SD), LDL (mean ± SD), HDL (mean ± SD).[HbA1C (%): (mean ± SD)] was replaced by [HbA1c (%)], and the mean HbA1c was removed from the table.


Their crystal structures exhibits alternating inorganic and organic stacking sheets or layers in (II), with Cl 6H22N4)[SnCl6]Cl2·2H2O, (I), and 1,4-bis(2-ammonioethyl)piperazin-1,4-diium hexachloridostannate (IV) dichloride dihydrate, (C8H24N4)[SnCl6]Cl2·2H2O, (II), have been synthesized from the same starting materials. In each case both the cations and anions are located about inversion centers. Their crystal structures exhibits alternating inorganic and organic stacking sheets in (I) and layers in (II), with Cl− ions and water molecules occupying the space in between. The cohesion of the three-dimensional frameworks are governed by N—H⋯Cl, N—H⋯O, C—H⋯Cl and O—H⋯Cl hydrogen bonds. Hirshfeld surface analysis of both crystal structures indicates that the H⋯Cl/Cl⋯H contacts exert an important influence on the stabilization of the packing.Two new organic–inorganic hybrid compounds, triethylenetetraammonium hexachloridostannate (IV) dichloride dihydrate, (C Piperazine derivatives are relatively more volatile than the corresponding linear polyethylene amines  and other branched or cyclic TETA, with close boiling points, such as tris-(2-aminoethyl)amine), 1,4-bis(2-aminoethyl)piperazine, (Bis AEP), and 2 salt. Under very mild reaction conditions, we believe that (Bis AEP) is present as an impurity in commercial TETA based on the fact that rearrangement reactions of aliphatic chelating polyamines require high pressure and temperature 4+ cation, one half of an inorganic [SnCl6]2- dianion, one Cl− ion and one molecule of water  dianion [SnCl6]2−, lying on a centre of inversion, exhibits a nearly perfect octahedral coordination sphere with Sn—Cl bond lengths ranging from 2.4114 (6) to 2.4469 (6) Å and Cl—Sn—Cl bond angles between 88.94 (2) and 91.06 (2)°.The asymmetric unit of (I)er Fig. 1. The 2−dianion lying on a centre of inversion  to 2.4331 (6) Å and Cl—Sn—Cl bond angles ranging from 88.55 (2) to 91.45 (2)° for the cis angles [180° for trans angles]. The organic part is totally protonated and the piperazinium portion adopts a chair conformation, with both ammonioethyl groups being in equatorial positions.The asymmetric unit of compound (II)on Fig. 2. The [Bi4+ and inorganic [SnCl6]2− sheets extending along the a-axis direction. The organic cations in adjacent chains are oriented in opposite directions, forming antiparallel sheets. The isolated chloride ions Cl− and the water molecules are located in the otherwise empty space between the sheets ts Fig. 3.W and C—H⋯Cl hydrogen-bonding interactions (Table 13+ group as well as the NH2+ group of [TETA]4+ act as hydrogen-bond donors. The D⋯A distances for the NH3+ group range from 2.980 (4) to 3.255 (3) Å, while D⋯A distances of 3.026 (2) to 3.452 (2) Å are found for the NH2+ group. The water molecules play an important role in stabilizing the crystal packing of (I)− anions and [SnCl6]2− dianions via O1W—H1W⋯Cl4 and O1W–-H2W⋯Cl2 hydrogen bonds with a H⋯Cl distances of 2.60 (5) and 2.82 (5) Å, respectively. Additionally, by playing the role of acceptors, the water molecules link the inorganic moieties with the organic cations through N1+—H1B⋯O1W and N1+—H1C⋯O1W charge-assisted hydrogen bonds with H⋯O distances of 2.09 and 2.25 Å, respectively.The crystal packing of (I)s Table 1. The NH34+ cations, are joined to their adjacent water molecules through strong OW—H⋯Cl hydrogen bonds, leading to a hydrogen-bonding pattern with a +—H1B⋯O1W and C6—H5B⋯Cl4 hydrogen bonds, promote the formation of sheets of cations aligned parallel to the  to 3.431 (2) Å and the C⋯Cl distances of 3.715 (3) Å.In (II)e Table 2. These sCrystalExplorer software In compounds (I)et al., 2016i.e. triethylenetetraminium bis(sulfate) monohydrate, (C6H22N4)SO4·H2O (III), and bis(2-ammonioethyl)piperazin-1,4-ium tetraperchlorate tetrahydrate, (C8H24N4)4ClO4·4H2O (IV), have been reported  and tetraethylenetetraamine in an HCl-acidified medium with a stoichiometric ratio of 1:1 was refluxed for one h at 343 K for (I)All chemicals were used without further purificationUiso(H) = 1.2Ueq. H atoms of the water molecule were located in a difference-Fourier map and refined with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989018001044/tx2003sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 1817660, 1817659CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The planar decachlorocyclopentasilane rings in the title compounds are coordinated by two chloride ions to generate inverse-sandwich complexes. n-butylammonium) dichloride decachlorocyclopentasilane dichloromethane disolvate, 2C16H36N+·2Cl−·Si5Cl10·2CH2Cl2, (I), and bis(tetraethylammonium) dichloride decachlorocyclopentasilane dichloromethane disolvate, 2C8H20N+·2Cl−·Si5Cl10·2CH2Cl2, (II), both of which crystallize with discrete cations, anions, and solvent molecules. In (I), the complete decachlorocyclopentasilane ring is generated by a crystallographic twofold rotation axis. In (II), one cation is located on a general position and the other two are disordered about centres of inversion. These are the first structures featuring the structural motif of a five-membered cyclopentasilane ring coordinated from both sides by a chloride ion. The extended structures of (I) and (II) feature numerous C—H⋯Cl interactions. In (II), the N atoms are located on centres of inversion and as a result, the ethylene chains are disordered over equally occupied orientations.We have determined the crystal structures of two decachlorocyclopentasilanes, namely bis(tetra- The distances of the chloride ions to the Si atoms  show that the chloride ions are located almost exactly above the centroid of the ring 2[Si5Cl12] (R = nBu or Et)  could be harvested after storage of the reaction solution for one week at 195 K in 89% and 93% yield, respectively. Both adducts are stable in the solid phase under inert conditions. However, in solution a rapid transformation of [nBu4N]2[Si5Cl12] to [nBu4N]2[Si6Cl14] and [nBu4N]2[Si7Cl16] , while [Et4N]2[Si5Cl12] is not soluble. For comparison, a 29Si CP/MAS NMR spectrum of single crystals of [nBu4N]2[Si5Cl12] was recorded .The addition of a solution of [t) Fig. 4. Crystal6] Fig. 5 can be omethyl—H = 0.98 Å or Cmethylene—H = 0.99 Å and with Uiso(H) = 1.5Ueq(Cmethyl) or 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 4The Cl atoms of the dichloromethane solvent molecule in (I)10.1107/S2056989017016310/hb7717sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989017016310/hb7717Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017016310/hb7717IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989017016310/hb7717sup4.pdfFig. S1. 29Si NMR spectra. DOI: 10.1107/S2056989017016310/hb7717sup5.pdfFig. S2. 29Si CP/MAS NMR spectrum. DOI: 1585194, 1585193CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


N,N′,N′′-donating imine ligand led to a mononuclear compound, the N,N′,O-donating imine ligand produced a binuclear metal complex.It is shown that tridentate imine ligands can control the nuclearity of copper(II) complexes based on the donor atoms present in the ligand. While the  N,N′,N′′-donating imine ligand led to a mononuclear compound, namely dichloridocopper(II) monohydrate, [CuCl2(C10H15N3)]·H2O, 1, while the N,N′,O-donating imine ligand produced a binuclear metal complex, namely μ2-chlorido-dichlorido(μ2-2-{[2-(dimethylamino)ethyl]iminomethyl}phenolato)dicopper(II) 0.11-hydrate, [Cu2(C11H15N2O)Cl3(C4H12N2)]·0.11H2O, 2. The structure of 2 is a remarkable example of a binuclear copper(II) complex containing a single substituted 2-iminomethylphenolate ligand that has two copper(II) sites in square-pyramidal coordination.It is demonstrated here that tridentate imine ligands can control the nuclearity of copper(II) complexes based on the donor atoms present in the ligand. The  Imines represent an interesting class of ligands because they can be easily synthesized and fine-tuned to the desired application by introducing extra donor atoms or groups with the desired steric properties into the side chains. A limited number of binuclear copper(II) compounds containing substituted 2-iminomethylphenole ligands have been reported in the literature  sites of haemocyanin  and binuclear (2) copper(II) complexes with tridentate imine-containing ligands obtained by a one-pot synthetic method. The nuclearity of the complexes was shown to be directed by the different donor atoms present in the imine ligand.We describe here the crystal structures of mononuclear . The CuII cation is displaced from the least-squares plane defined by the four coordinating atoms of the square base  by 0.334 Å. The bond lengths to these atoms are: Cu—N1 = 2.060 (2), Cu—N2 = 1.978 (2), Cu—N3 = 2.058 (2) and Cu—Cl2 = 2.2639 (8) Å; the Cu—Cl bond length to the apical Cl1 atom that completes the first coordination sphere is considerably longer, at 2.5013 (8) Å. In order to assess the coordination geometry of copper(II) more quantitatively, the τ5 index as defined by Addison et al.  cations, both in a square-pyramidal coordination environment . The presence of the phenolate group in the structure of the imine ligand directs the reaction with copper(II) cations to form a binuclear coordination compound, in contrast with the mononuclear species 1 obtained when a pyridine group is present in the ligand. Atoms Cu1 and Cu2 in 2 are displaced from the least-squares plane defined by the four coordinating atoms of the square base  by 0.299 and 0.170 Å, respectively. The distances from the central copper(II) cations to these ligating atoms are: Cu1—N1 = 2.068 (2), Cu1—N2 = 1.959 (2), Cu1—O1 = 1.968 (1) and Cu1—Cl2 = 2.2958 (5) Å; Cu2—N3 = 2.021 (2), Cu2—N4 = 2.040 (2), Cu2—Cl3 = 2.2501 (5) and Cu2—O1 = 2.004 (1) Å. The two Cu—Cl distances to the apical Cl atoms are likewise longer, Cu1—Cl1 = 2.5476 (5) Å and Cu2—Cl2 = 2.5938 (5) Å. The Cu⋯Cu distance within the binuclear complex is 3.2525 (5) Å. In compound 2, the τ5 index for Cu1 is 0.294 and for Cu2 0.260, indicating more distorted square-pyramidal coordination environments for both central copper(II) cations.The binuclear compound nt Fig. 2a. The p2, a solvent-accessible void of 42 Å3 was detected by a PLATON analysis  distance of 3.393 (2) Å . The partly occupied water molecule participates in a hydrogen bond with the μ2-bridging Cl2 ligand phenolate ligand were considered as analogues of 2. A total of 12 hits were found as analogues of 1, while 11 hits were found for analogues of 2, including both mono- and bis(iminomethyl)phenolate ligands. Averages of selected bond lengths  and the statistical analysis module in Mercury (Version 3.9)  compounds 1 is associated with the nonhydrated analogue , Cu—N2 = 2.104 (2) and Cu—N3 = 2.236 (2) Å, and almost identical Cu—Cl1 = 2.2573 (5) and Cu—Cl2 = 2.22561 (6) Å distances. The CuII cation is displaced from the mean plane defined by the four coordinating atoms of the square base by 0.622 Å. While for 1 τ5 = 0.0593, for the structure of TAWMEK τ5 = 0.302. The differences in the coordination environment of copper(II) probably arise as a consequence of the presence of the hydrogen-bonded network established between the chloride ligands and the water molecules in the crystal structure of 1. The coordination spheres around the CuII cations in 1 and TAWMEK are compared in Fig. 4The closest relation to 2, the search returned only two examples of binuclear CuII complexes containing a single μ2-(monoiminomethyl)phenolate ligand [VAMJIE  complex with a single μ2-(monoiminomethyl)phenolate ligand that has two CuII coordination sites in square-pyramidal environments. The coordination spheres around the two CuII cations in 2 and UFATEB are compared in Fig. 4Regarding the binuclear compound N,N-dimethylethylenediamine, pyridine-2-carboxaldehyde and salicylaldehyde were purchased from Sigma–Aldrich and used without further purification.Copper(II) chloride dihydrate was purchased from Vetec (Brazil). 1, C10H15Cl2CuN3·H2O, was obtained as follows. In a 10 ml beaker, N,N-dimethylethylenediamine  was combined with pyridine-2-carboxaldehyde  in methanol (200 µl). The reaction was carried out at room temperature for 24 h. Afterwards, solid CuCl2·2H2O  was added to the reaction mixture. A polycrystalline green compound was obtained, filtered off and washed with small amounts of cold methanol. Elemental analysis was performed on a Perkin–Elmer CHNS-O 2400. Analysis, calculated for C10H15Cl2CuN3·H2O: C 36.4, H 5.2, N 12.7%; found: C 36.8, H 4.8, N 13.0%. The supernatant was transferred to an amber flask and green crystals suitable for single-crystal X-ray diffraction were obtained by slow evaporation.Compound 2 (C15H27Cl3Cu2N4O·0.11H2O) was obtained following the same synthetic procedure as used for 1, but replacing pyridine-2-carboxaldehyde by salicylaldehyde (11 µl). Green needle-like crystals of 2 were obtained by slow evaporation of the supernatant. Since only a few crystals were obtained, no further analytical data were acquired.Compound 2) or 0.95 Å (CH), with Uiso(H) = 1.2Ueq(C), and C—H = 0.98 Å (CH3) and Uiso(H) = 1.5Ueq(C). For structure 1, the H atoms of the water molecule were refined with an O—H distance restraint of 0.82 (1) Å and a H⋯H separation of 1.29 (2) Å, and with Uiso(H) = 1.5Ueq(O). For structure 2, the H atoms of the amine functionality (H3A and H3B) were refined freely. The occupancy of the partly occupied water solvent molecule was refined to a value of 0.11 (1); for this molecule, H atoms were not located and they were not considered in the final model.Crystal data, data collection and structure refinement details are summarized in Table 410.1107/S2056989017013652/wm5417sup1.cifCrystal structure: contains datablock(s) global, 2, 1. DOI: 10.1107/S2056989017013652/wm54171sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989017013652/wm54172sup3.hklStructure factors: contains datablock(s) 2. DOI: 1576100, 1576099CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


H-tetrazol-1-olate) dianion.In energetic materials, the crystal density is an important parameter that affects the performance of the material. When making ionic energetic materials, the choice of counter-ion can have detrimental or beneficial effects on the packing, and therefore the density, of the resulting energetic crystal. Presented herein are a series of five ionic energetic crystals, all containing the 5,5′-bis(1 H-tetrazol-1-olate), with the following cations: hydrazinium (1) (2N2H5+·C6N12O42−), hydroxylammonium (2) 2NH4O+·C6N12O42− , dimethylammonium (3) (2C2H8N+·C6N12O42−), 5-amino-1H-tetrazol-4-ium (4) (2CH4N5+·C6N12O42−·4H2O), and aminoguanidinium (5) (2CH7N4+·C6N12O42−). Both the supramolecular interactions and the sterics of the cation play a role in the density of the resulting crystals, which range from 1.544 to 1.873 Mg m−1. In 5, the tetrazolate ring is disordered over two positions [occupancy ratio 0.907 (5):0.093 (5)] due to a 180° rotation in the terminal tetrazole rings.In energetic materials, the crystal density is an important parameter that affects the performance of the material. When making ionic energetic materials, the choice of counter-ion can have detrimental or beneficial effects on the packing, and therefore the density, of the resulting energetic crystal. Presented herein are a series of five ionic energetic crystals, all containing the dianion 5,5′-bis(1 As a result of the variety of cation structures and intermolecular interactions, each exhibits subtly different crystal packing, which affects the resulting density. The molecule of interest, however, only exhibits minor changes in bond distances depending on the cation.One of the critical parameters directly related to the performance of an energetic material, specifically its detonation velocity, is the density of the material (Ma H-tetrazol-1-olate), is comprised of four pentanuclear rings, with two 1,2,4-oxadiazole rings linked together through the 5-position carbon atom, and the tetrazol-1-olate rings linked at the 5-position carbon atom to each 1,2,4-oxadiazole ring at the 3-position carbon.The primary molecule, 5,5′-bis%] in which one tetrazolate is flipped by 180°. The N4—C5—C6—N10 torsion angles for 1 [174.25 (13)°], 4 , and 5  show only slight deflections from coplanar, while in 2 [168.63 (15)°], the deflection is more pronounced. In structure 3, the N4—C5—C6—N10 dihedral angle is 2.38 (19)°, showing only a slight deviation from coplanarity, despite the proximity of the two electronegative oxygen atoms.In each structure, the oxadiazole oxygen atoms are on opposite sides. For 4 Fig. 4, with thet al., 1987et al., 1987In all five structures, the tetrazolate C—N and N—N bond distances  suggest a delocalized aromatic system rather than discrete single and double bonds  Å matches the distance of 1.45 Å seen in hydrazinium chloride  Å matches the distance of 1.41 Å seen for hydroxylammonium perchlorate  and 1.4780 (17) Å are consistent, albeit on the low side, with those reported for dialkylammonium ions, on average 1.494 (16) Å ]: C—Namino, 1.320 (2) and 1.314 (2) Å (1.308 Å); C—Nring, 1.334 (2) to 1.338 (2) Å (1.334 to 1.342 Å); C—N(H)—N=N, 1.357 (2) to 1.366 (2) Å (1.363 to 1.366 Å); N(H)—N=N—N(H), 1.272 (2) and 1.269 (2) Å ]: C—NH2, 1.309 (3) and 1.320 (3) Å (1.312 and 1.320 Å); C—N(H)(NH2), 1.337 (3) Å (1.328 Å); and N(H)—NH2, 1.420 (3) Å bis(1H-tetrazol-1-olate) (dianion) with the N2—N3 bond of one dianion over the tetrazolate ring of the dianion in the neighboring column . Hydrazinium ions occupy the gaps between neighboring coplanar dianions along the b-axis, above the plane of the molecules. One hydrazinium forms a hydrogen-bonded network linking the neighboring intrasheet dianions through the tetrazolate oxygen, tetrazolate N4 atom, and the NH3 portion of hydrazinium. Additionally, hydrogen bonds form between the NH2 portion of hydrazinium, the tetrazolate oxygen atom, and the tetrazolate N3 atom of neighboring dianions. An additional hydrogen bond connects the NH3 of one hydrazinium with the NH2 portion of the symmetry-related hydrazinium ion  and 4.01 (2) Å. The tetrazolate oxygen atom forms an anion–π interaction with the oxadiazole ring of a neighboring dianion, with an O1-to-centroidC6/O7/N8/C9/N10 close contact of 2.98 (2) Å at an O1–centroidC6/O7/N8/C9/N10–O7 angle of 92.3 (2)° . Additionally, the opposing columns are staggered with respect to one another. The hydroxylammonium cations occupy the space formed where three dianion columns meet, above the dianion planes. The arrangement of the dianions in the peaks and troughs of the packing is dictated by the hydrogen bonds between the hydroxylammonium hydroxyl group and the tetrazolate oxygen atom, and those between the hydroxylammonium NH3 group and O1, N2, and N4 of three symmetry-related dianions  Å, centroidN1–N4/C5–centroidC6/O7/N8/C9/N10–O7 angle, 80 (2)°] interaction between the tetrazolate and oxadiazole rings. Additionally, the tetrazolate oxygen atom does not participate in an anion–π interaction with the oxadiazole ring due to the stronger π–π interaction. The oxadiazole rings of neighboring dianions are far apart, at a centroidC6/O7/N8/C9/N10–centroidC6/O7/N8/C9/N10 distance of 4.26 (2) Å and a centroidC6/O7/N8/C9/N10–centroidC6/O7/N8/C9/N10–N10 angle of 50 (2)°, suggesting minimal π–π interaction.Structure ns Fig. 7a. Addit3, space group Pa), with the dianion stacked in a staggered arrangement, with the tetrazolate ring of one dianion over the central oxadiazole–oxadiazole C—C bond of the dianions above and below. The oxadiazole ring resides over the tetrazolate–oxadiazole C—C bond in the dianions above and below. The void space between the dianion columns is occupied by dimethylammonium ions, located within the plane of the molecules in an up–down arrangement. Two dimethylammonium ions are positioned between the sheets, forming hydrogen bonds between the NH2 group and the tetrazolate oxygen atoms of dianions in neighboring sheets  and 3.99 (2) Å (the latter distance to the inversion-related oxadiazole of the same dianion).Structure rn Fig. 8a, with 4, space group P21/c, packs in the sheet-like pattern consisting of extended sheets containing the dianion, cations, and incorporated water . The 5-amino-1H-tetrazol-4-ium cations and water molecules surround each dianion, isolating the dianion from other dianions within the sheets. Between the sheets, the dianion only interacts with another dianion via one terminal tetrazolate ring, with the oxygen atom of the tetrazolate over the C—C bond between the tetrazolate and oxadiazole rings. Within each sheet, there is extensive hydrogen bonding between the dianions, 5-amino-1H-tetrazol-4-ium, and incorporated water molecules, isolating the dianions from one another in the sheet plane  Å [N–-centroidN1–N4/C5–centroidN1–N4/C5 angle 62.0 (2)°]. The C11-oxadiazole engages in a π–π interaction with its symmetry equivalent as well, at a centroidC11/N12/O13/C14/N15–centroidC11/N12/O13/C14/N15 distance of 3.93 (2) Å . A π–π interaction is also seen between the N30-tetrazolium ring and its symmetry equivalent, at a centroidC29/N30–N33–centroidC29/N30–N33 distance of 3.69 (2) Å . Additionally, there are two anion–π interactions, the first between O21 and the N1-tetrazolate of a neighboring dianion, and the second between O21 and the C6-oxadiazole, with an O21–centroidN1–N4/C5 distance of 3.33 (2) Å [O21–centroidN1–N4/C5–N2 angle 95.8 (2)°] and O21–centroidC6/O7/N8/C9/N10 3.02 (2) Å [O21–centroidC6/O7/N8/C9/N10–C6 angle 76.3 (2)°].Structure er Fig. 9a. The 55, space group P21/n, packs in a mixing pattern, with columns containing stacked sheets consisting of the dianion coplanar with two aminoguanidinium cations . Neighboring columns of sheets are rotated by 67° with respect to one another as a result of the hydrogen bonding of the amino group of the cation with the oxygen atom of a neighboring oxadiazole ring. In fact, it is the hydrogen-bonding interaction between the amino group of the aminoguanidinium cation and the oxygen atom of the oxadiazole that directs the mixing-type packing seen in the crystal structure. The planar portion of the aminoguanidinium cation interacts via hydrogen bonds from the unsubstituted guanidinium amines to the tetrazolate oxygen atom, oxadiazole N8, and symmetry-related oxadiazole N10 atoms of one dianion, and to the tetrazolate N2 atom of a neighboring dianion  Å [centroidC6/O7/N8/C9/N10–centroidA–N4A/C5AN1–N1A angle 65.4 (2)°].Structure s Fig. 10a. Neigh3 < 6 < 1 < 5 < 2. Unsurprisingly, the dimethylammonium, with minimal hydrogen bonding, non-interacting substituents, and a poor steric match for the dianion, is the least dense of the structures shown. Aminoguandinium, despite significant hydrogen bonding, exhibits a lower density as well, likely due to the directionality of the hydrogen-bond donors, which directs packing of the dianions into less efficient arrangements. Hydrazinium benefits from extensive hydrogen bonding, but the orientation of the hydrazinium directs the dianions into slightly less efficient packing than the hydroxylammonium cation, preventing the staggering of the columns that allows for improved space occupation. The 5-amino-1H-tetrazol-4-ium cation, with the second-highest density, packs very efficiently, in extended sheets with extensive hydrogen bonding, losing out to the hydroxylammonium cation likely only due to the included water molecules needed to fill in gaps between the dianions and cations. Hydroxylammonium exhibits the most efficient, highest-density packing due to the directing influence and strong hydrogen-bond donating ability of the hydroxyl group, which forms a short hydrogen bond and directs the columns into a staggered arrangement, fitting the dianions slightly closer together at the point where neighboring columns meet. The range of densities, from 1.544 to 1.873 g cm−1, shows the significant effect that matching the hydrogen-bonding abilities and sterics of the counter-ion to the primary energetic ion has on efficient packing and, by extension, the expected performance of these ionic energetics.As demonstrated above, it is the hydrogen-bonding networks that establish the crystal packing exhibited in each example, with π–π and anion–π interactions occurring if packing allows. As shown in Table 6et al., 2016H-tetrazol-1-olate). A search using 5-[3-]-1H-tetrazolate also yields no results. Searching for the ring fragments separately yielded 443 structures for 1,2,4-oxadiazole and 127 structures for tetrazol-1-olate. The closest structures to those presented herein are dimers between similar ring fragments. A search for each of the cations yields the following results: 196 structures containing hydrazinium, 99 structures containing hydroxylammonium, 1,583 structures containing dimethylammonium, 2,230 structures containing ammonium, 17 structures containing 5-amino-1H-tetrazol-4-ium, and 130 structures containing aminoguanidinium.A search of the CSD -5,5′-dichloroxime (6) and 5,5′--5,5′-diyl)bis(1-hydroxytetrazole) (7), have been described previously  was added to a 20 ml vial with water (1.5 ml) and a stir bar. Hydrazine hydrate  was added to the reaction mixture and heated until dissolved. Stirring was discontinued, the stir bar was removed, and the solution was allowed to cool slowly providing crystals of 1.Compound 3: In a round-bottom flask, fitted with a drying tube, was suspended chloroxime 6  in dimethylformamide (DMF) , which was then cooled in an ice–water bath. Sodium azide  was added in portions with stirring, and the reaction was allowed to warm to room temperature. Additional DMF (10 ml) was added to the creamy mixture, and after 1.5 h, the solids went into solution. At this point, complete formation of the diazidoxime was assumed, and cyclization to 1 proceeded as follows. A 1:1 mixture of diethyl ether/dioxane was added to the reaction mixture , and the solution was cooled to 273 K with an ice bath. HBr or Cl2 gas was bubbled into the reaction at which time the temperature increased to 298 K. Gas was added until the reaction temperature returned to approximately 278 K, and the vessel was then stoppered and allowed to stir for 22 h. The voluminous, white precipitate that formed (hygroscopic dimethylamonium bromide) was separated by vacuum filtration, and the filtrate was allowed to evaporate from a crystallizing dish. Upon evaporation, a white solid (3) in a yellow oil remained. The solid was separated from the oil by vacuum filtration (535 mg). 3 was crystallized by heating in minimal water and slow cooling.Compound 4: Dihydrate 7  was added to a 20 ml vial with water (1.5 ml) and a stir bar. 5-Aminotetrazole  was added to the mixture, which was then heated with stirring until dissolved. Stirring was discontinued, the stir bar was removed, and the solution was allowed to cool slowly providing crystals of 4.Compound 5: Dihydrate 7  was added to a 20 ml vial with water (1.5 ml) and a stir bar. Aminoguanidinium H2CO3  was added to the mixture, which was then heated with stirring until dissolved. During dissolution, gas evolved, the solution became clear, followed by the formation of a tan precipitate. Heating was continued until complete dissolution, followed the removal of the stir bar, and slow cooling to provide crystals of 5.Compound 5, the tetrazolate ring  is disordered over two positons (A and B) due to a 180° rotation in the terminal tetrazole rings. The disorder has the relative ratio of 90.7 (5):9.3 (5). CCDC deposition numbers are as follows: 1, CCDC 1567779; 2, CCDC 1567780; 3, CCDC 1567783; 4, CCDC 1567784; 5, CCDC 1567804.Crystal data, data collection and structure refinement details are summarized in Table 710.1107/S205698901800364X/pk2606sup1.cifCrystal structure: contains datablock(s) global, 1, 2, 3, 4, 5. DOI: 10.1107/S205698901800364X/pk26061sup9.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S205698901800364X/pk26062sup10.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S205698901800364X/pk26063sup15.hklStructure factors: contains datablock(s) 3. DOI: 10.1107/S205698901800364X/pk26064sup20.hklStructure factors: contains datablock(s) 4. DOI: 10.1107/S205698901800364X/pk26065sup8.hklStructure factors: contains datablock(s) 5. DOI: Click here for additional data file.10.1107/S205698901800364X/pk26061sup7.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901800364X/pk26062sup8.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901800364X/pk26063sup9.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901800364X/pk26064sup10.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901800364X/pk26065sup11.cmlSupporting information file. DOI: 1567804, 1567784, 1567783, 1567780, 1567779CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


II complex with a heterocyclic pyrazole ligand, a potential candidate for metal catalysts, are reported. The CuII atom is coordinated by three O atoms and two N atoms, provided by a tridentate pyridine-2,6-dicarboxylate, one pyrazole and one water ligand, forming a slightly distorted square-pyramidal geometry.The synthesis and crystal structure of tridentate pyridine-2,6-dicarboxylate Cu 7H3NO4)(C3H4N2)(H2O)]·2H2O, the CuII atom is coordinated by three O atoms and two N atoms, provided by a tridentate pyridine-2,6-dicarboxylate (pdc), one pyrazole and one water ligand, forming a slightly distorted square-pyramidal geometry [range of O—Cu—O and O—Cu—N bond angles = 79.55 (8)–166.22 (10)°]. The water molecule is positioned at the apical position. In the crystal, the complex molecule and the two crystallographically independent non-coordinating water molecules are linked into a supramolecular layer structure parallel to the ab plane via O—H⋯O and N—H⋯O hydrogen bonds.In the title compound, [Cu(C H-pyrazol-1-yl)methyl]pyridine are known to be catalysts of polyethylene polymerization methyl]pyridine was oxidized to pyridine-2,6-dicarboxylate (pdc) by metal nitrate , pyrazole and water ligands. The coordination geometry around the CuII atom is a distorted square pyramid as indicated by the τ value of 0.113  and two oxygen atoms (O9 and O12 from pdc). The plane including the CuII atom is almost planar, with an r.m.s. deviation of 0.0847 Å from the corresponding least-squares plane defined by the five constituent atoms. The pyrazole ring is twisted by 66.61 (10)° from the basal plane. The apical Cu1—O19 bond length of 2.217 (2) Å is much longer than those of the basal Cu—O lengths [Cu1—O9 = 2.026 (2) Å and Cu1—O12 = 2.058 (2) Å].The molecular structure of the title compound is shown in Fig. 1B⋯O21, O20—H20B⋯O13 and O20—H20A⋯O10iii; symmetry code as in Table 1A⋯O20iv and O21—H21B⋯O20v; Table 1i and O19—H19A⋯O9ii; Table 1ab plane methyl]pyridine  in acetonitrile (5 ml) in a high-pressure vessel. After sealing the high-pressure vessel, the resulting solution was stirred for three days at 403 K. The precipitate formed was removed by filtration, and the filtrate was washed with acetonitrile and dichloromethane to get a dark-green powder. Single crystals of the title compound were obtained from its aqueous solution by slow evaporation of the solvent at 333 K within five days.A solution of copper nitrate trihydrate  in acetonitrile (5 ml) was added to a solution of 2,6-bis[(1Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989017016231/is5480sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017016231/is5480Isup2.hklStructure factors: contains datablock(s) I. DOI: 1584872CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


II cation, located on a twofold rotation axis, is coordinated by two 2,4,6-trimethylbenzoate anions, two nicotinamide ligands and a water molecule in a distorted pentagonal–bipyramidal geometry.The Cd 10H11O2)2(C6H6N2O)2(H2O)], contains one half of the complex molecule, with the CdII cation and the coordinated water O atom residing on a twofold rotation axis. The CdII cation is coordinated in a bidentate manner to the carboxylate O atoms of the two symmetry-related 2,4,6-trimethylbenzoate (TMB) anions and to the water O atom at distances of 2.297 (2), 2.527 (2) and 2.306 (3) Å to form a distorted pentagonal arrangement, while the distorted pentagonal–bipyramidal coordination sphere is completed by the two pyridine N atoms of the two symmetry-related monodentate nicotinamide (NA) ligands at distances of 2.371 (3) Å in the axial positions. In the crystal, molecules are linked via intermolecular N—H⋯O, O—H⋯O and C—H⋯O hydrogen bonds with R22(12), R33(8), R33(14), R33(16), R33(20), R33(22), R44(22), R55(16), R66(16) and R66(18) ring motifs, forming a three-dimensional architecture. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are H⋯H (56.9%), H⋯C/C⋯H (21.3%) and H⋯O/O⋯H (19.0%) interactions.The asymmetric unit of the title complex, [Cd(C A deficiency of this vitamin leads to loss of copper from the body, known as pellagra disease. Victims of pellagra show unusually high serum and urinary copper levels 2(C6H6N2O)2]  ·H2O   , O1—Cd1—O2 [53.63 (7)°], O1i—Cd1—O2i [53.63 (7)°], O2—Cd1—O4 [84.47 (5)°] and O2i—Cd1—O4 [84.47 (5)°] in the basal plane around CdII cation is 363.77° . This confirms the presence of the CdII cation with a small deviation from the basal plane. The distorted pentagonal–bipyramidal coordination sphere is completed by the two pyridine N atoms (N1 and N1i) of the two symmetry-related monodentate nicotinamide (NA) ligands at distances of 2.371 (3) Å in the axial positions ° in (II), 59.02 (8)° in (IV), 58.45 (9)° in (V) and 60.70 (4)° in (VI). In the TMB anion, the carboxylate group is twisted away from the attached benzene ring, A (C2–C7), ring by 60.94 (18)°, while the benzene and pyridine rings [pyridine = B (N1/C11–C15)], are oriented at a dihedral angle of 50.32 (11)°. The four-membered ring D (Cd1/O1/O2/C1) is nearly planar with a maximum deviation of 0.0029 (30) Å (for C1) from the mean plane, and it is oriented at dihedral angles of 60.98 (11) and 81.91 (7)°, with respect to the A and B rings.The near equalities of the C1—O1 [1.249 (4) Å] and C1—O2 [1.253 (3) Å] bonds in the carboxylate groups indicate delocalized bonding arrangements, rather than localized single and double bonds. The O2—C1—O1 bond angle [121.7 (3)°] seems to be slightly decreased than that present in a free acid [122.2°]. The O2—C1—O1 bond angle may be compared with the corresponding values of 123.5 (2) and 120.4 (2)° in (II), 125.2 (5)° in (III), 119.2 (3) and 123.8 (2)° in (IV), 123.6 (3) and 119.4 (3)° in (V) and 123.86 (13) and 118.49 (14)° in (VI), where the benzoate ions are coordinated to the metal atoms only monodentately in (III), and both monodentately and bidentately in (II), (IV), (V) and (VI). The Cd1 atom lies 0.0192 (1) Å above of the planar (O1/O2/C1) carboxylate group. The O1—Cd1—O2 angle is 53.63 (7)°. The corresponding O—via intermolecular N—HNA⋯ONA, N—HNA⋯OC, O—HW⋯ONA and C—HTMB⋯OC  hydrogen bonds  analysis  in water (50 ml) and nicotinamide  in water (25 ml) with sodium 2,4,6-trimethylbenzoate  in water (150 ml) at room temperature. The mixture was filtered and set aside to crystallize at ambient temperature for six weeks, giving colourless single crystals . Combustion analysis: found; C, 57.07, H, 5.67, N, 7.92%. Calculated: C32H36CdN4O7 C, 57.42; H, 5.43, N, 8.34%. FT–IR: 3390, 3122, 2921, 1669, 1619, 1539, 1445, 1399, 1113, 1038, 847, 731, 641 cm−1.The title compound was prepared by the reaction of 3CdSO2 group and of the water molecule were located in difference-Fourier maps and refined freely. The C-bound H atoms were positioned geometrically with C—H = 0.93 and 0.96 Å for aromatic and methyl H atoms, respectively, and constrained to ride on their parent atoms, with Uiso(H) = k × Ueq(C), where k = 1.5 for methyl H-atoms and k = 1.2 for aromatic H-atoms.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989018001494/xu5916sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018001494/xu5916Isup2.hklStructure factors: contains datablock(s) I. DOI: 1818756CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Individual molecules are stacked along the b-axis direction. The cohesion in the crystal structure is accomplished by C—H⋯F hydrogen bonds and additional off-set π–π interactions , leading to the formation of a three-dimensional supramolecular network.The molecule of the title 1,8-naphthyridine-BF The individual F—B bond length is 1.364 (5) Å and the Br—C bond length 1.940 (4) Å. Compared with the molecular structure of a related compound  Å, interplanar distance = 3.6085 (1) Å, slippage = 0.472 Å; Cg2 is the centroid of the N2/C3–C6/C11 ring; symmetry code: (i) −x, −y, 2 − z] ethanimidato](difluoro)borate  was added dropwise to an ice-cooled solution of 2,6-lutidine (1 ml) and N-acetamide  in anhydrous CH2Cl2 (80 ml) under a nitrogen atmosphere. After the mixture had been stirred for 24 h under ambient temperature, the reaction was quenched with 20 ml distilled water. The aqueous layer was extracted with CH2Cl2 (3 × 50 ml); the organic layer was dried with anhydrous Na2SO4 and the solvent removed under reduced pressure. The residue was purified by silica gel chromatography using CH2Cl2 as eluent to give the pure product as a bright white powder . Yellow crystals of the title compound were obtained from its CH2Cl2 solution by slow evaporation at room temperature.BFUiso(H) = 1.2Ueq(C) for aromatic and Uiso(H) = 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989016016704/wm5325sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016016704/wm5325Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016016704/wm5325Isup3.cmlSupporting information file. DOI: 1510400CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:


II–NaI metal–organic framework has been synthesized and the X-ray structure has been reported.A novel three-dimensional bimetallic Cd 2(C8H4O4)2(C3H7NO)(H2O)2]n or [CdNa22(DMF)(H2O)2]n, is a new CdII–NaI heterobimetallic coordination polymer. The asymmetric unit consists of one CdII atom, two NaI atoms, two 1,3-bdc ligands, two coordinated water molecules and one coordinated DMF molecule. The CdII atom exhibits a seven-coordinate geometry, while the NaI atoms can be considered to be pentacoordinate. The metal ions and their symmetry-related equivalents are connected via chelating–bridging carboxylate groups of the 1,3-bdc ligands to generate a three-dimensional framework. In the crystal, there are classical O—H⋯O hydrogen bonds involving the coordinated water molecules and the 1,3-bdc carboxylate groups and π–π stacking between the benzene rings of the 1,3-bdc ligands present within the frameworks.The title compound, [CdNa Studies incorporating alkaline metal ions into d10-MOFs with one type of bridging ligand to construct novel heterobimetallic d10-alkaline metal ion MOFs have been undertaken ] where 1,2,4-btc = benzene-1,2,4-tricarboxylate 3·(DMF)2·(m-H2O)2] where 1,4-bdcH2 = benzene-1,4-dicarboxylic acid 2]} where 1,3-bdcH2 = benzene-1,3-dicarboxylic acid 2]·2H2O where OH-1,3-bdcH2 = 5-hydroxybenzene-1,3-dicarboxylic acid 6(H2O)8] where ntcH3 = 5-nitrobenzene-1,2,3-tricarboxylic acid , we explored mixed sources of ZnII/CdII–NaI with this ligand. The expected products are prepared by using a direct synthetic method, mixing metal nitrate salts, 1,3-bdcH2 and NaOH (mole ratio 1:1:2) in water, methanol and DMF solvents. However, only the CdII–NaI MOF product has been successfully synthesized. As part of our ongoing studies on this complex, we describe here the synthesis and crystal structure of a novel three-dimensional heterobimetallic CdII–NaI MOF, [CdNa22(DMF)(H2O)2]n (I).Porous coordination polymers or metal–organic frameworks (MOFs) constructed from I) crystallizes in the tetragonal crystal system with polar P43 space group. The asymmetric unit of (I) consists of one CdII ion, two crystallographically independent Na(I) ions, two 1,3-bdc ligands, two coordinated water molecules and one DMF molecules, as shown in Fig. 1II ion is coordinated by seven carboxylate oxygen atoms from four different 1,3-bdc ligands with the Cd—O bond distances range between 2.301 (3) and 2.555 (3) Å (Table 1I). The 1,3-bdc molecule is fully deprotonated and coordinated to the CdII and NaI ions in a μ5-coordination mode, creating a one-dimensional heterobimetallic chain running parallel to the c axis, Fig. 3a- and b-axis directions, generating a three-dimensional framework structure as shown in Fig. 4The title compound (Å Table 1. The Na1I), classical O—H⋯O hydrogen bonds and aromatic π–π stacking interactions are observed and these interactions presumably help to stabilize the frameworks. All water molecules are shown to act as O—H⋯O hydrogen-bond donors towards the carboxylate groups of the 1,3-bdc ligands (Table 2Cg1⋯Cg2i distance of 3.588 (3) Å and a dihedral angle of 3.8 (4)° .In the crystal of (s Table 2. The π–πI), only the three-dimensional coordination framework {[CdNa2]·[NH2(CH3)2]} has been reported 2 molecules. In comparison, compound (I) contains coordinated H2O and DMF molecules projecting into the framework channels. Other related three-dimensional heterobimetallic d10–NaI coordination frameworks containing benzenepolycarboxylate ligands have been published, such as [CdNa2(H2O)2]·2H2O where OH-1,3-bdcH2 = 5-hydroxy-benzene-1,3-dicarboxylic acid 3·(DMF)2·(m-H2O)2] where 1,4-bdcH2 = benzene-1,4-dicarboxylic acid 6(H2O)8] where ntcH3 = 5-nitrobenzene-1,2,3-tricarboxylic acid  and NaOH (2.0 mmol) in 10 mL of distilled water was slowly dropped to a methanolic solution (10 ml) of Cd(NO3)2·4H2O (1.0 mmol). The reaction mixture was stirred at 333 K for 30 min and allowed to cool to room temperature and then filtered. The filtrate was allowed to stand to slowly evaporate at ambient temperature. Colorless block-shaped crystals suitable for single crystal X-ray diffraction were obtained after three days (76% yield based on Cd).A mixture solution of 1,3-bdcHUiso(H) = 1.2Ueq(C). The coordinated DMF molecule was found to be disordered with two sets of sites with a refined occupancy ratio of 0.382 (10) and 0.618 (10).Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989017013871/pj2046sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017013871/pj2046Isup2.hklStructure factors: contains datablock(s) I. DOI: 1576543CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Despite a growing appreciation of γδ T cell contributions to numerous immune responses, the mechanisms that underpin their thymic development remain poorly understood. Here, using precursor/product relationships, we identify thymic stages in two distinct developmental pathways that generate γδ T cells pre-committed to subsequent secretion of either IL-17A or IFNγ. Importantly, this framework for tracking γδ T cell development has permitted definitive assessment of TCRγδ signal strength in commitment to γδ T cell effector fate; increased TCRγδ signal strength profoundly prohibited the development of all IL-17A-secreting γδ T cells, regardless of Vγ usage, but promoted the development of γδ progenitors along the IFNγ pathway. This clarifies the recently debated role of TCRγδ signal strength in commitment to distinct γδ T cell effector fates and proposes an alternate methodology for the study of γδ T cell development.  •CD44 and CD45RB identify two distinct thymic γδ T cell developmental pathways•Cytokine-secretion-independent identification of effector fate-committed γδ T cells•Sizable numbers of IL-17A-committed γδ T cells express Vγ1 and Vγ2/3 chains•Increased TCRγδ signal strength prohibits development of IL-17A-secreting γδ T cells Sumaria et al. identify distinct thymic pathways that generate murine γδ T cells pre-committed to the secretion of IL-17A or IFNγ. This permits assessment of TCRγδ signal strength in thymic commitment to γδ T cell effector fate; increased TCRγδ signal strength profoundly prohibits development of all IL-17A-secreting γδ T cells. Anti-tu−CD8− double-negative (DN) 2 cells (rather than DN3 cells) (+ cells) may inherently require certain transcription factors   was suggested to promote commitment to an IFNγ-secreting fate , with we3 cells) . And cer Sox-13) . ClearlyThere is presently no accepted approach for stage-wise assessment of thymic γδ T cell development. Indeed, although studies have analyzed Vγ usage , acquisi− cells that expressed high CD44 but not CD45RB were committed to IL-17A secretion, but did not make IFNγ, whereas cells that had upregulated CD45RB had potential to secrete IFNγ but not IL-17A  that re-capitulates thymic T cell development in vitro and is suited to studying γδ T cell development that occurs predominantly in the perinatal period. Indeed, E15 thymic lobes cultured for 7 days generate γδ T cell subsets similar to those observed ex vivo -purified the four CD24 d cells C. Sortedectively C and 2D.products C and 2D,+ and Vγ6+ cells being linked to IL-17A production, with Vγ1+ and Vγ5+ cells linked to IFNγ  expression in Vγ6+ cells  cells of making IL-17A  express T-bet but not RORγt. Interestingly, our CD44/CD45RB plots show overlap with CD44/Ly-6C plots suggested to identify naive-like and memory-like peripheral γδ T cell subsets  supports a model in which commitment to an IL-17A- or IFNγ-secreting fate, with initial expression of corresponding “master” transcriptional regulators . These observations suggest thymic commitment of γδ progenitors to distinct effector fates is distinguishable  from actual capacity to secrete cytokine.Importantly, our analyses identify two thymic pathways of functional γδ T cell differentiation that diverge from a common CD24gulators , spans a+Vδ1+ cells in the presence of Skint1  γδ T cells in SKG mice that have severely reduced Zap-70 activity  effects of increased TCRγδ signal strength mediated by anti-TCRδ antibody. Thus, activation of the ERK/MAP kinase pathway by strong TCRγδ signaling is a key limiter of progression to an IL-17A-secreting fate. As mentioned above, such strong signaling may reflect engagement of TCR ligand, as supported by complete segregation, in the prenatal thymus, of Vγ5 pathway . Howeverignaling  that depnsidered . These iAdditional details are available in C57BL/6 (B6) mice were purchased from Charles River Laboratories. All mice were fetal (E14–E17), neonatal (1–3 days), post-natal (4–8 days), or adult . All experiments involving animals were performed in full compliance with UK Home Office regulations and institutional guidelines.Thymic lobes from B6 mice were cultured on Nuclepore membrane filter discs (Whatman) in complete RPMI-1640 medium plus 10% fetal calf serum (FCS) for 7–14 days.OP9-DL1 cells were provided by J.C. Zúniga-Pflücker (University of Toronto).For detection of Vγ5Vδ1 and Vγ6Vδ1, cells were pre-stained with GL3 followed by 17D1. For i.c. cytokine staining (eBioscience), cells were stimulated with 50 ng/ml phorbol 12-myristate 13-acetate  and 1 μg/ml ionomycin (Sigma) for 4 hr at 37°C. Acquisition was performed with an LSR-II or a Canto II (BD). Analysis was performed using FlowJo (Tree Star).GraphPad Prism software was used to analyze data, which are presented as mean ± SD. Two-tailed Student’s unpaired t test was used when only two groups were compared, and one-way ANOVA with Tukey’s test was used for multiple comparisons. Significance was determined at p ≤ 0.05.N.S. and C.L.G. performed experiments. N.S. and D.J.P. analyzed the data. B.S.-S. and D.J.P. designed the study. D.J.P. and N.S. wrote the paper.


III and DyIII with the ligand 2,6-diacetylpyridine bis(benzoylhydrazone) have been synthesized and structurally characterized.Two isotypic complexes of Tb Ln(DAPBH2)(CH3OH)(H2O)3]Cl3·2CH3OH , are isotypic. The central lanthanide ions are nine-coordinate, being ligated by three N and two O atoms from the pentadentate DAPBH2 ligand, and four O atoms from the coordinated methanol molecule and three coordinated water molecules. The coordination geometry of the lanthanide ion is a distorted capped square antiprism. In the crystals, the various components are linked by O—H⋯Cl, N—H⋯Cl and O—H⋯O hydrogen bonds, forming three-dimensional supramolecular frameworks. Within the frameworks, there are C—H⋯Cl and C—H⋯O hydrogen bonds and offset π–π interactions (intercentroid distance ca 3.81 Å).The title lanthanide complexes, [ The bond distances for the DyIII complex are slightly shorter than those of the TbIII complex as a result of the lanthanide contraction effect. The DAPBH2 ligand is approximately planar, and the LnIII ion lies out of the mean plane (O1/N2/N3/N4/O2) by a distance of 0.5754 (3) Å for the TbIII complex and 0.5702 (3) Å for the DyIII complex. The coordination of the DAPBH2 ligand to the lanthanide ion shows a bent arrangement . These coordination features are similar to those reported for the dysprosium DAPBH2 nitrate complex  Å, α = 3.8 (1)°, interplanar distance = 3.483 (1) Å, slippages = 1.77 and 1.55 Å; Cg2 and Cg3 are the centroids of C2–C7 and C18–C23 rings, respectively, symmetry codes: (a) x, y − 1, z; (b) x, y + 1, z]. The layers are linked by O—H⋯O, O—H⋯Cl and N—H⋯Cl hydrogen bonds, forming a three-dimensional supramolecular framework, which is reinforced by a series of C—H⋯Cl and C—H⋯O hydrogen bonds , 2,6-diacetylpyridine , and benzoylhydrazine  was refluxed for 2 h. The resulting mixture was filtered. Vapour diffusion of diethyl ether into the filtrate afforded colourless plate-like crystals of the TbIII complex . The synthetic procedure for the DyIII complex is similar, starting from dysprosium chloride (yield 43%).A methanol solution (15 ml) of TbClIII complex was restrained to 0.82 Å. Other hydrogen atoms were generated geometrically and refined with a riding model: N—H = 0.88 Å, C–H = 0.95–0.98 Å with Uiso(H) = 1.5Ueq and 1.2 Ueq for other H atoms.Crystal data, data collection, and structure refinement details are summarized in Table 410.1107/S2056989018004103/su5429sup1.cifCrystal structure: contains datablock(s) TbDAPBH2, DyDAPBH2, global. DOI: 10.1107/S2056989018004103/su5429TbDAPBH2sup2.hklStructure factors: contains datablock(s) TbDAPBH2. DOI: 10.1107/S2056989018004103/su5429DyDAPBH2sup3.hklStructure factors: contains datablock(s) DyDAPBH2. DOI: 1828810, 1828809CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The organic molecule is twisted with a near orthogonal relationship between the outer rings [dihedral angle = 71.79 (6)°]. Supramolecular ribbons sustained by hydrogen bonding feature in the molecular packing. 14H12BrN3O2·H2O {systematic name: N′-[(1E)-1-(5-bromo-2-hydroxyphenyl)ethylidene]pyridine-4-carbohydrazide monohydrate}, the central CN2O region of the organic molecule is planar and the conformation about the imine-C=N bond is E. While an intramolecular hydroxy-O—H⋯N(imine) hydrogen bond is evident, the dihedral angle between the central residue and the benzene rings is 48.99 (9)°. Overall, the molecule is twisted, as seen in the dihedral angle of 71.79 (6)° between the outer rings. In the crystal, hydrogen-bonding interactions, i.e. hydrazide-N—H⋯O(water), water-O—H⋯O(carbonyl) and water-O—H⋯N(pyridyl), lead to supramolecular ribbons along the a-axis direction. Connections between these, leading to a three-dimensional architecture, are mediated by Br⋯Br halogen bonding [3.5366 (3) Å], pyridyl-C—H⋯O(carbonyl) as well as weak π–π interactions [inter-centroid separation between benzene rings = 3.9315 (12) Å]. The Hirshfeld surface analysis reveals the importance of hydrogen atoms in the supramolecular connectivity as well as the influence of the Br⋯Br halogen bonding.In the title isonicotinohydrazide hydrate, C Recent interest in the coordination of hydrazide Schiff base ligands arises owing to the presence of electron-donating nitrogen and oxygen atoms, allowing these to act as a multidentate ligands, and in some cases, function as supramolecular building blocks in their molecular assemblies °], N2—N1—C1—C10 [179.59 (15)°] and C1—N1—N2—C2 [171.14 (18)°] torsion angles is consistent with an all-trans relationship in the central chain and a small twist about the N1—N2 bond. The conformation about the imine-C2=N2 bond [1.292 (2) Å] is E. An intramolecular hydroxy-O1—H⋯N2(imine) hydrogen bond is noted, Table 1The molecular structures of the constituents of (I)a-axis direction and mediated by hydrogen-bonding interactions. In essence, the water molecule provides links between three organic molecules via hydrazide-N—H⋯O(water), water-O—H⋯O(carbonyl) and water-O—H⋯N(pyridyl) hydrogen bonds, Table 14O}2 synthons as shown in Fig. 2a. Lateral connections between ribbons are via halogen bonding of the type Br⋯Br. Here, the Br⋯Bri separation is 3.5366 (3) Å . The C7—Br⋯Bri angle is 156.56 (5)°, and, being disposed about a centre of inversion, the C7—Br⋯Bri—C7i torsion angle is constrained by symmetry to 180°. The geometric characteristics indicate the Br⋯Bri halogen bond is classified as a type I halogen bond  within the surface near the water-O1W atoms as well as the blue regions  about the water-H1W and H2W atoms, which correspond to the acceptor and donors of the hydrogen bonds, respectively. Similarly, the other donor and acceptor atoms participating in the more significant intermolecular interactions are viewed as the blue and red regions, respectively, in Fig. 3dnorm in Fig. 4b also highlight the significant contribution of Br⋯Br and C⋯C contacts to the molecular packing. The presence of faint-red spots near the pyridyl-N3, C13 and H13 atoms and the carbonyl-O1 atom indicate their contributions to short interatomic C⋯N/N⋯C contacts  and shape-index-  mapped Hirshfeld surfaces highlighting the various short interatomic contacts influential on the molecular packing are illustrated in Fig. 5a, C⋯C and C⋯N/N⋯C contacts, Fig. 5b, and Br⋯Br and Br⋯H/H⋯Br contacts, Fig. 5c, identify their roles in consolidating the packing in the crystal.The analysis of the Hirshfeld surface for (I)s Table 2 and compm- Fig. 5a and b x- Fig. 5c mappeda, and those delineated into H⋯H, O⋯H/H⋯O, C⋯H/H⋯C, Br⋯H/H⋯Br, N⋯H/H⋯N, C⋯C, Br⋯Br and C⋯N/N⋯C contacts  hydrogen bonds are seen on the dnorm-mapped Hirshfeld surface in Fig. 4b). The conformational relationship between each of the pyridyl and benzene rings to the central planar region make these residues available for forming C⋯H/H⋯C contacts. The significant contribution of 17.9% from C⋯H/H⋯C contacts results from the short interatomic contacts listed in Table 2d with the pair of peaks at de + di ∼ 2.8 Å; these short interatomic contacts are illustrated in Fig. 5a. A forceps-like fingerprint plot corresponding to Br⋯H/H⋯Br contacts in Fig. 6e with its tips at de + di ∼ 3.0 Å represents the influence of the halogen⋯hydrogen interactions in the molecular packing. Along with Br⋯H/H⋯Br contacts, Table 2via Br⋯Br contacts, as evident in Fig. 6h as a very thin line beginning at de + di ∼ 3.5 Å. The contributions from other interatomic contacts involving the bromide atom have negligible effect on the crystal stability because their interatomic distances are much greater than sum of their respective van der Waals radii. The small but notable contributions from the C⋯C and C⋯N/N⋯C contacts to the Hirshfeld surface, Table 2g, a spear-shaped distribution of points with the tip at de + di ∼ 3.2 Å and an adjacent arrow-like distribution of points at de = di ∼ 1.9 Å result, respectively, from interatomic C⋯C contacts and π–π stacking interactions involving the C3–C8 ring. The short interatomic C⋯N/N⋯C contacts involving the pyridyl-C13 and N3 atoms, Fig. 5b, are reflected in a pair of small peaks at de + di ∼ 3.2 Å in Fig. 6i. The small contributions from other interatomic contacts listed in Table 2The overall two-dimensional fingerprint plot, Fig. 6et al., 2016et al., 2014cf. the near to orthogonal relationship in (I)The most closely related structure to (I)−1. The 1H NMR spectrum was recorded at room temperature in CDCl3 solution on a Jeol ECA 400 MHz FT–NMR spectrometer.All chemicals and solvents were used as purchased without purification, and all reactions were carried out under ambient conditions. The melting point was determined using an Electrothermal digital melting-point apparatus and was uncorrected. The IR spectrum for the compound was obtained on a Perkin Elmer Spectrum 400 FT Mid-IR/Far-IR spectrophotometer from 4000 to 400 cmn-butyltin dichloride  in methanol (10 ml). The resulting mixture was stirred and refluxed for 3 h. A cloudy orange solution was obtained and the mixture was filtered. The filtrate was allowed to stand at room temperature and yellow crystals, suitable for X-ray crystallographic studies, were obtained after the slow evaporation. The yellow crystals were found to be a side-product from the reaction mixture. Yield: 0.112 g, 34%; M.p. 501 K. IR (cm−1): 3158(br), 1666(s), 1548(s), 1152 (m), 964(s) cm−1. 1H NMR (in CDCl3): 11.20 , 8.73-8.82, 7.92-8.20, 6.80-6.99 , 4.82 , 4.10 , 3.13 .1-(5-Bromo-2-hydroxyphenyl)ethylidene]isonicotinohydrazide  and triethylamine  in methanol (25 ml) were added to di-Uiso(H) set to 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 510.1107/S2056989017004790/hb7669sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017004790/hb7669Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017004790/hb7669Isup3.cmlSupporting information file. DOI: 1540550CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The title compound is an epimer of natural tetracyclic isosaxorumamide. The dihydrofuranone, oxolane, azepane and pyrrolidine rings adopt planar, twist, twist-chair and envelope forms, respectively. In the crystal, O—H⋯O hydrogen bonds connect the water and main molecules into a tape structure, which is further expanded into a three-dimensional network by C—H⋯O interactions. 17H23NO4·H2O, is an epimer of the natural tetracyclic alkaloid isosaxorumamide which consists of a fused 5–7–5 tricyclic core and a dihydrofuranone substituent. The terminal dihydrofuran ring is essentially planar with a maximum deviation of 0.0273 (14) Å from the mean plane and oxolane, azepane and pyrrolidine rings in the tricyclic ring system adopt twist, twist-chair and envelope forms, respectively. In the crystal, the amide and water molecules are linked by O—H⋯O hydrogen bonds, forming a tape structure running along the b-axis direction. The tapes are further connected by C—H⋯O interactions into a three-dimensional architecture.The title compound, C Stemona alkaloids isolated from the root of Stemona Saxorum  = 0.350 (3) Å and φ(2) = 271.0 (4)°. Atoms C8 and C9 deviate from the plane through the other three atoms by 0.309 (6) and −0.271 (6) Å, respectively. The central seven-membered azepane ring (N1/C2–C5/C9/C10), which is trans-fused to the oxolane ring, adopts a twist-chair form with puckering parameters of Q = 0.796 (2), Q(2) = 0.472 (2) Å, φ(2) = 195.0 (3)°, Q(3) = 0.641 (2) Å and φ(3) = 246.7 (2)°. The pyrrolidine ring (N1/C10–C13) fused to the azepane ring adopts an envelope form with puckering parameters of Q(2) = 0.300 (3) Å and φ(2) = 251.1 (5)°. The flap atom C11 deviates from the mean plane through the other four atoms by 0.473 (4) Å. The amide moiety (N1/C2/C10/C13/O14) is planar, and atoms N1 and O14 deviate from the mean plane through the other three atoms by 0.028 (2) and −0.035 (4) Å, respectively.The asymmetric unit of the title compound is shown in Fig. 2W—H1WA⋯O14i and O1W—H1WB⋯O14ii; symmetry codes as in Table 1b-axis direction. A C—H⋯O interaction  graph-set motif. Furthermore, a weak C—H⋯O interaction , i.e. the fused tricyclic core related to the title compound , with CSD refcodes VATJAC , with refcodes PROTMI  is the natural alkaloid (–)-stemoamide, which is a -7,13-dione derivative of (a), and XATFOP (space group P21/n) is its racemate.In the Cambridge Structural Database . HRMS (ESI) m/z calculated for C17H24NO4+ [M + H]+: 306.1705; found: 306.1703.The title compound was afforded in a synthetic study of saxorumamide and isosaxorumamide, from ethyl 4-bromobutanoate and a siloxypyrrole analogue (Yoritate Uiso(H) = 1.2Ueq(C) or 1.5Ueq(methyl C). Water H atoms were located in a difference-Fourier map, and then refined freely with Uiso(H) = 1.5Ueq(O), and with distance restraints of O—H = 0.84 (2) Å and H⋯H = 1.33 (4) Å.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989018004425/is5494sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018004425/is5494Isup2.hklStructure factors: contains datablock(s) I. DOI: 1830268CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


N-(2-iodophenyl)acetamide into twofold interwoven sheets, and the molecules of 2-(4-chlorophenyl)-N-(pyrazin-2-yl)acetamide are linked into complex sheets built solely from hydrogen bonds.A combination of N—H⋯O and C—H⋯O hydrogen bonds together with C—Cl⋯π(arene) and C—I⋯π(arene) interactions links the molecules of 2-(4-chlorophenyl)- N-(2-iodophenyl)acetamide, C14H11ClINO, molecules are linked by a combination of N—H⋯O and C—H⋯O hydrogen bonds to form a C(4)C(4)[R21(7)] chain of rings and chains of this type are linked by a combination of C—Cl⋯π(arene) and C—I⋯π(arene) interactions to form deeply puckered twofold interwoven sheets. In the crystal of 2-(4-chlorophenyl)-N-(pyrazin-2-yl)acetamide, C12H10ClN3O, molecules are linked into complex sheets by N—H⋯N, C—H⋯N and C—H⋯O hydrogen bonds, and by C—H⋯π(arene) interactions.In the crystal of 2-(4-chlorophenyl)- R1CH2CONHR2, where R1 and R2 are aromatic or hetero-aromatic substituents, are of interest as they have some resemblance to benzyl penicillins ca 0.01 Å, indicating that this ring is fully aromatic.In the pyrazine ring of compound (II)C(4)C(4) direction  x, y, z, where Cg1 represents the centroid of the C11–C16] ring, I⋯Cg 3.7977 (14), C⋯Cg 5.082 (3) Å and C—I⋯Cg 116.34 (8)°; for C24—Cl24⋯Cg2ii , Cl⋯Cg 3.4557 (8), C⋯Cg 4.504 (3) Å and C—Cl⋯Cg 116.19 (11)°. The metrics of the C—Cl⋯Cg interaction are well within the normal range, as deduced using database analysis on Fig. 3. There ion Fig. 3, lying won Fig. 3. The dimi 3.058 (2) Å and C12—I12⋯O1i 170.88 (8)°  which complements the C—Cl⋯Cg contact. The I⋯O distance here is significantly shorter than the sum of the van der Waals radii, 3.56 Å C(5) for the group of 428 very weak reflections having Fc/Fc(max) in the range 0.000 < Fc/Fc(max) < 0.008.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989016012512/su5316sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989016012512/su5316Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016012512/su5316IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989016012512/su5316Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016012512/su5316IIsup5.cmlSupporting information file. DOI: 1497360, 1497359CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


N,S-chelating hydrazinecarbodithioate ligands provide a trans-N2S2 donor set and a distorted square-planar geometry for the NiII atom. In the crystal, a three-dimensional network is sustained by C—H⋯π and π–π interactions.Two  19H21N2S2)2], is generated by the application of a centre of inversion. The NiII atom is N,S-chelated by two hydrazinecarbodithioate ligands, which provide a trans-N2S2 donor set that defines a distorted square-planar geometry. The conformation of the five-membered chelate ring is an envelope with the NiII atom being the flap atom. In the crystal, p-tolyl-C—H⋯π(benzene-iPr), iPr-C—H⋯π(p-tolyl) and π–π interactions [between p-tolyl rings with inter-centroid distance = 3.8051 (12) Å] help to consolidate the three-dimensional architecture. The analysis of the Hirshfeld surface confirms the importance of H-atom contacts in establishing the packing.The complete molecule of the title hydrazine carbodithioate complex, [Ni(C S-R-dithiocarbazate (R = methyl/benzyl/methylbenzyl) and heterocyclic aldehydes or ketones have received much attention in recent years owing to their cytotoxicity S,N-chelating hydrazinecarbodithioate anions. From symmetry, the resulting N2S2 donor set has like atoms trans, and the square-planar coordination geometry is strictly planar. Distortions from the ideal geometry are related to the deviations of angles subtended at nickel by the donor atoms, Table 1i.e. −165.61 (17)°. Despite being involved in a formal bond to the NiII atom, the C1—S1 bond length of 1.7296 (19) Å is still significantly shorter than those formed by the S2 atom, i.e. C1—S2 = 1.7479 (18) Å and C12—S2 = 1.824 (2) Å.The Ni2S2 donor set does not extend to the five-membered chelate ring, which has an envelope conformation with the nickel atom lying 0.465 (2) Å above the least-squares plane through the remaining atoms [r.m.s. deviation = 0.0016 Å]. The sequence of C1=N1, N1—N2 and N2=C2 bond lengths of 1.294 (2), 1.408 (2) and 1.300 (2) Å, respectively, suggests limited conjugation across this residue. Each of the benzene rings of the S- and N-bound substituents is twisted with respect to the least-squares plane through the chelate ring. Thus, a nearly orthogonal relationship exists between the chelate and p-tolyl rings, with the dihedral angle being 89.72 (5)°. Less dramatic is the twist of the iPr-substituted ring with the dihedral angle being 13.83 (9)°. The dihedral angle between the aromatic rings is 84.31 (6)°.The planarity of the Ni.e. the S1 and N1 atoms, are involved in intramolecular interactions, Table 2i.e. p-tolyl-C—H⋯π(iPr-benzene) and iPr-benzene-C—H⋯π(p-tolyl) contacts, Table 2p-tolyl rings self-associate via face-to-face, π–π, interactions , indicating the p-tolyl ring participates in two distinct interactions. The result of the supramolecular association is the formation of a three-dimensional architecture, Fig. 2The two sites potentially available for hydrogen bonding in (I)et al., 2017dnorm (not shown) clearly indicates the absence of conventional hydrogen bonding in the crystal. The donors and acceptors of C—H⋯π interactions, involving atoms of each of the iPr-benzene and p-tolyl rings, are viewed as blue and light-red regions and correspond to the respective positive and negative potentials on the Hirshfeld surface mapped over electrostatic potential (over the range ± 0.025 au), Fig. 3iPr-benzene and p-tolyl rings on the Hirshfeld surface mapped over de, Fig. 4i.e. π⋯H—C contacts, between iPr–H11B and the p-tolyl ring are represented by red and white dotted lines, respectively in Fig. 5a; the blue dotted lines in Fig. 5a represent π–π stacking between p-tolyl rings at −x, −1 − y, 1 − z. The other C—H⋯π contacts involving p-tolyl-H17 and iPr-benzene rings are illustrated in Fig. 5b.The Hirshfeld surface analysis for (I)et al., 2007a–f. From the quantitative summary of the relative contributions of the various interatomic contacts given in Table 3i.e. 95.3%. In the fingerprint plot delineated into H⋯H contacts. Fig. 6b, the points are distributed in the major part of the plot, but they do not make significant contributions to the molecular packing as their interatomic separations are greater than sum of their van der Waals radii, i.e. de + di > 2.4 Å. The presence of short interatomic C⋯H/H⋯C contacts, see Table 4i.e. 22.2%. This is consistent with the fingerprint plot, Fig. 6c, where the short interatomic C⋯H/H⋯C contacts appear as a pair of small peaks at de + di ∼ 2.8 Å and also as the blue regions around the participating hydrogen atoms, namely H5 and H10B, on the Hirshfeld surface mapped over electrostatic potential, Fig. 3d and e, indicate meaningful contributions to the Hirshfeld surface, Table 3i.e. 2.1%, but recognizable contribution from C⋯C contacts to the Hirshfeld surface is ascribed to π–π stacking interactions between symmetry-related p-tolyl rings, and appear as an arrow-like distribution of points around de = di 1.9 Å in Fig. 6f. The other contacts have low percentage contributions to the surface and are likely to have negligible effects on the molecular packing, Table 3The overall two-dimensional fingerprint plot and those delineated into H⋯H, C⋯H/H⋯C, S⋯H/H⋯S and N⋯H/H⋯N and C⋯C contacts , with a formal link between the two imine functionalities, the cis-N2S2 arrangement is imposed by the geometric requirements of the bis(dithiocarbazate) di-anion  dithiocarbazate complexes in the crystallographic literature (Groom S-4-methylbenzyldithiocarbazate (S4MDTC) precursor was synthesized by following a procedure adapted from the literature : 1589, ν(C=N); 997, ν(N—N); 823, ν(C=S).The synthesized Schiff base  was dissolved in hot acetonitrile (50 ml) and added to nickel(II) acetate tetrahydrate  in an ethanolic solution (30 ml). The mixture was heated and stirred to reduce the volume of the solution. Precipitation occurred once the mixture cooled to room temperature. The precipitate then was filtered and dried over silica gel. The complex was recrystallized from its methanol solution. Brown prismatic crystals were formed from the filtrate after being left to stand for a month. The crystals were filtered and washed with absolute ethanol at room temperature. Yield: 70%. M.p.: 479–480 K. Elemental composition calculated for CUiso(H) set to 1.2–1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 510.1107/S2056989017002419/hb7658sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017002419/hb7658Isup2.hklStructure factors: contains datablock(s) I. DOI: 1532446CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information: