Examination of the morphology of these materials indicates that SPT aggregates were reduced when CNCs or [C2mim][OAc] were present. Inclusion of CNCs and/or SPT had a greater effect on material properties when the matrices were un-plasticised. Addition of SPT or CNCs altered the crystalline structure of the un-plasticised chitosan matrix. Moreover, a combination of SPT and CNCs was more effective at suppressing re-crystallisation. Nonetheless, the mechanical properties and surface hydrophobicity were more related to CNC/SPT–biopolymer interactions. The un-plasticised bionanocomposites generally showed increased relaxation temperatures, enhanced tensile strength, and reduced surface wettability. For the [C2mim][OAc] plasticised matrices, the ionic liquid (IL) dominates the interactions with the biopolymers such that the effect of the nanofillers is diminished. However, for the [C2mim][OAc] plasticised chitosan/CMC matrix, CNCs and SPT acted synergistically suppressing re-crystallisation but resulting in increased tensile strength.Cellulose nanocrystals (CNCs) and/or sepiolite (SPT) were thermomechanically mixed with un-plasticised chitosan and chitosan/carboxymethyl cellulose (CMC) blends plasticised with 1-ethyl-3-methylimidazolium acetate ([C Deionised water was used in all experiments.Chitosan -chitosan  and the chitosan . CNCs we2mim][OAc] at 20% (“E2”) of the matrix. The suffix “F” indicates the processed samples were in film form. A-F and B-F (without plasticiser or nano-additives) [2mim][OAc] at 20% but without nano-additives) [OAc], such band shifts obtained on inclusion of CNCs were not apparent, suggesting the IL disrupted the interactions between the CNCs and the biopolymers. Furthermore, regardless of matrix type and plasticiser, no significant changes to the bands were observed with the inclusion of SPT, as the interaction of SPT with the biopolymers was weaker than with the CNCs.linkage) ,35,36, ilinkage) ,35,36. Fetching) ,35,36. Tθ position  and the peak at 27.2° ((110) reflection, 0.38 nm) moved to a lower 2θ position . Thus, inclusion of CNCs or SPT at a rather low content (0.75 wt%) largely impacted the packing of chitosan chains and suppressed the re-crystallisation of chitosan. In contrast to A/C-F and A/S-F, A/CS-F displayed an XRD curve that matches that for A-F, where the peak intensities were even weaker. In this regard, there may be interaction between CNCs and SPT, which combined results in greater steric hindrance limiting chain movement for re-crystallisation. Compared with AE2-F [OAc] (Td = 252 °C) reduced the thermal stability of chitosan [OAc] led to a more defined peak and the overlapping peak at the higher temperature was significantly diminished, ascribed to the enhanced mixing and interactions between the two biopolymers due to the presence of the IL [OAc] plasticised A-matrix, inclusion of CNCs and/or SPT did not result in changes to both Tα and Tβ. For AE2/S-F, AE2/C-F, and AE2/CS-F, the mobility of either side or main chains should be mainly determined by the plasticiser. for A-F  with a β for A-F ,38. For Tα = −43 °C and Tβ = 97 °C) [OAc], Tα (−23 °C) and Tβ (91 °C) did not vary on inclusion of CNCs or SPT, whereas for BE2/CS-F, Tα (−18 °C) and Tβ (97 °C) increased slightly. This indicates that, while the IL aided biopolymer chain mobility for the B-matrix, CNCs and SPT combined to reduce the chain mobility of the plasticised biopolymers. Compared with B-F (= 97 °C) , B/S-F, The stress–strain curves from tensile testing  indicateE), tensile strength (σt), and elongation at break (εb) of all the materials were calculated and plotted in εb than A-F (22.6 ± 4.6%) [OAc], inclusion of CNCs and/or SPT did not cause significant changes in mechanical properties of the matrix. In these samples, the hydrogen-bonded network in chitosan was significantly weakened by addition of the IL, and further inclusion of the nanofillers did not impact this plasticisation state, behaviour supported by the FTIR, XRD, and DMTA results.From the stress–strain curves, the Young’s modulus ( ± 4.6%) , indicatE = 1325 ± 176 MPa, σt = 50.5 ± 3.6 MPa, and εb = 10.4 ± 3.4%) [OAc] plasticised A-matrix, inclusion of CNCs and/or SPT had no significant effect on θc0s and θc60s. In this case, the surface hydrophilicity was mainly determined by the interactions between chitosan and [C2mim][OAc] and these interactions were on the whole not affected by CNCs or SPT.asing θc . Compare68 ± 5°) , A/S-F aθc0s = 82 ± 5° and θc60s = 65 ± 7° for B/S-F, θc0s = 83 ± 5° and θc60s = 73 ± 7° for B/C-F, and θc0s = 88 ± 4° and θc60s = 76 ± 6° for B/CS-F in contrast to θc0s = 71 ± 6° and θc60s = 60 ± 5° for B-F [OAc], the CNCs or SPT may have interacted with the IL, thus reducing the overall material hydrophilicity. However, BE2/CS-F displayed θc0s and θc60s similar to those of BE2-F. Again, interaction between CNCs and SPT reduces their respective interactions with the IL or the biopolymers.For the un-plasticised B-matrix, inclusion of CNCs and/or SPT resulted in increased surface hydrophilicity ( for B-F ). It is 2mim][OAc]. Furthermore, the inclusion of CNCs and/or SPT was shown to largely impact the biopolymer crystallinity. Specifically, A/S-F and A/C-F showed a different XRD pattern indicating that SPT or CNCs alter the packing of chitosan chains to form crystals. The combination of SPT and CNCs showed a synergistic effect, more effective at suppressing chitosan re-crystallisation, as seen for A/CS-F, B/CS-F, and BE2/CS-F. However, the material properties were not dependent on crystallinity but more related to the SPT/CNC–biopolymer interactions. While inclusion of CNCs and/or SPT apparently suppressed the crystallinity of the un-plasticised A- and B-matrices, the un-plasticised bionanocomposites generally showed increased relaxation temperatures, enhanced mechanical properties (both σt and E for the A-series and only σt for the B-series), and reduced surface wettability. In particular, B/S-F had a Tβ = 126 °C, θc0s = 100 ± 4°, and θc60s = 92 ± 5°; the σt values of A/CS-F and B/CS-F were 55.5 ± 0.8 MPa and 56.9 ± 3.2 MPa, respectively; and θc0s = 88 ± 4° and θc60s = 76 ± 6° for B/CS-F. For the [C2mim][OAc] plasticised matrices, the IL dominated the interactions with the biopolymers so that the effect of the nanofillers became weaker. Thus, AE2/S-F, AE2/C-F, and AE2/CS-F had unchanged properties. However, for the IL-plasticised B-matrix, there was a synergistic effect of CNCs and SPT on the biopolymer hydrogen bonding and electrostatic interactions and, thus, mechanical properties (BE2/CS-F had a σt = 47.6 ± 4.4 MPa). Hence, this work demonstrates the importance of tailoring the competing interactions in biopolymer nanocomposite systems for achieving desirable properties.Examination of the morphology of these bionanocomposites indicated that the extent of SPT aggregation could be reduced by inclusion of [C


Mthn⋯OBnzthz hydrogen bonds and are linked into chains extending along the a-axis direction by weak C—HBnz⋯OBnzthz  hydrogen bonds.The benzothiazine moiety is folded along the N⋯S axis and a puckering analysis of the conformation of the heterocyclic ring was performed. The hexyl chain is mainly in an extended conformation. In the crystal, inversion dimers are formed by weak C—H 22H25NOS, consists of methylbenzylidene and benzothiazine units linked to a hexyl moiety, where the thiazine ring adopts a screw-boat conformation. In the crystal, inversion dimers are formed by weak C—HMthn⋯OBnzthz hydrogen bonds and are linked into chains extending along the a-axis direction by weak C—HBnz⋯OBnzthz  hydrogen bonds. A Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H⋯H (59.2%) and H⋯C/C⋯H (27.9%) interactions. Hydrogen bonding and van der Waals interactions are the dominant interactions in the crystal packing. Computational chemistry indicates that in the crystal, the C—HBnz⋯OBnzthz and C—HMthn⋯OBnzthz hydrogen-bond energies are 75.3 and 56.5 kJ mol−1, respectively. Density functional theory (DFT) optimized structures at the B3LYP/ 6–311 G level are compared with the experimentally determined molecular structure in the solid state. The HOMO—LUMO behaviour was elucidated to determine the energy gap. Moreover, the antibacterial activity of the title compound was evaluated against gram-positive and gram-negative bacteria.The title compound, C We report herein the synthesis, the molecular and crystal structures along with the Hirshfeld surface analysis and interaction energy calculation [using CE–B3LYP/6–31G energy model] and the density functional theory (DFT) computational calculation carried out at the B3LYP/6–311 G level for comparing with the experimentally determined molecular structure in the solid state of the title compound. Moreover, the antibacterial activity of I is evaluated against gram-positive and gram-negative bacteria .In a continuation of our research devoted to the development of substituted 1,4-benzothiazine derivatives , ring conformation gave the parameters QT = 0.4853 (12) Å, θ = 69.48 (15)° and φ = 329.03 (18)°, indicating a screw-boat conformation. The dihedral angle between the benzene rings A (C1–C6) and C (C16–C21) is 75.64 (5)°. The base of the n-hexyl chain is approximately perpendicular to the mean plane of the benzothiazine unit, as indicated by the C6—N1—C9—C10 torsion angle of −96.2 (1)°. The remainder of this chain, with the exception of the terminal methyl group, is in an extended conformation  analysis  has the tips at de + di = 2.77 Å. The pair of spikes in the fingerprint plot delineated into H⋯S/S⋯H  has the tips at de + di = 2.98 Å. The H⋯O/O⋯H contacts  have a symmetrical distribution of points with the tips at de + di = 2.27 Å. Finally, the H⋯N/N⋯H contacts , make only a 0.8% contribution to the HS with the tips at de + di = 3.28 Å.The overall two-dimensional fingerprint plot, Fig. 7ts Fig. 7f, make dnorm plotted onto the surface are shown for the H⋯H, H⋯C/C⋯H, H⋯S/S⋯H and H⋯O/O⋯H interactions in Fig. 8a--d, respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H⋯H and H⋯C/C⋯H interactions suggest that van der Waals interactions and hydrogen bonding play the major roles in the crystal packing  is the sum of electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies  are −15.5 (Eele), −2.9 (Epol), −109.6 (Edis), 62.8 (Erep) and −75.3 (Etot) for C4—H4⋯O1 and −24.8 (Eele), −9.3 (Epol), −60.1 (Edis), 46.9 (Erep) and −56.5 (Etot) for C15—H15⋯O1.The intermolecular interaction energies are calculated using CE–B3LYP/6–31G energy model available in via density functional theory (DFT) using standard B3LYP functional and 6–311 G basis-set calculations , hardness (η), potential (μ), electrophilicity (ω) and softness (σ) are recorded in Table 3σ is for the evaluation of both the reactivity and stability. The electron transition from the HOMO to the LUMO energy level is shown in Fig. 9H-benzo[b]thiazin-3(4H)-one ring. The energy band gap [ΔE = ELUMO − EHOMO] of the molecule is 4.0189 eV, and the frontier molecular orbital energies, EHOMO and ELUMO are −5.8458 and −1.8269 eV, respectively.The optimized structure of the title compound in the gas phase was generated theoretically et al., 2016IIR1 = Ph, R2 = C), gave 15 hits, including with R1 = 4-ClC6H4 and R2 = CH2CH2CH2CH3 8CH3  , we have tested Pseudomonas aeruginosa (ATCC-27853), Staphylococcus aureus (ATCC-25923) and Streptococcus fasciens (ATCC-29212) strains of bacteria using the diffusion method disk for evaluating the applicability of I as an antibacterial agent -3,4-dihydro-2H-1,4-benzothiazin-3-one showed increases of MIC = 20 µg ml−1 for Staphylococcus aureus, MIC = 10 µg ml−1 for Escherichia coli and Pseudomonas aeruginosa and MIC = 5 µg ml−1 for Streptococcus fasciens, which corresponds to the best MIC activity as compared to the commercial antibiotic. In addition, the maximum effect of I was recorded against Pseudomonas aeruginosa (diameter of inhibition 12.1 mm). Chlor presents an antibacterial activity diameter of inhibition of between 19 mm and 27 mm and no zone inhibition was observed with dimethylsulfoxide (DMSO) [(1%): 1 mL of DMSO added to 99 mL ofulltra-pure water]  On one hand, the chemical structure of I can explain this biological effect. The mechanism of action of I is not attributable to one specific mechanism, but there are several targets in the cell: degradation of the cell wall, damage to membrane proteins, damage to cytoplasmic membrane, leakage of cell contents and coagulation of cytoplasm. On the other hand, it should be noted that the functionalized derivatives by ester groups and benzene rings have the highest antibacterial coefficient (92% of pathogenic bacteria are sensitive). This study is expected to take anti-inflammatory, antifungal, anti-parasitic and anti-cancer activities, because the literature gives a lot of interesting results on these topics. Some other types of bacteria may possibly be tested by employing the same method so as eventually to generalize the suggested investigation method , H-1,4-benzothiazin-3-one , potassium carbonate (4 mmol) and tetra-n-butyl ammonium bromide (0.2 mmol) in DMF (15 ml) was added 1-chlorohexane . Stirring was continued at room temperature for 12 h. The reaction mixture was filtered and the solvent was removed. The residue was extracted with water. The organic compound was chromatographed on a column of silica gel using the mixture ethyl acetate–hexane (9:1) as eluent. Colourless crystals of the title compound ITo a solution of 2-(4-methylbenzylidene)-3,4-dihydro-21H NMR  δ ppm: 0.88 ; 2.37 ; 2.37–2.52 ; 4.08 ; 7.06–7.57 ; 7.77 ; 13C NMR  δ ppm: 13.86 (–CH2–CH3); 20.98 (=CH–C6H4–CH3); 22.02, 25,83 26.48, 30.84, (CH2); 44.24 (NCH2); 117.26, 123.47, 126.36, 127.55, 129.15, 129.15, 130.03, 130.03, (CHarom); 133.77 ; 118.5, 119.31, 131.39, 135.77, 139.92, (Cq); 160.48 (C=O).Uiso(H) = k × Ueq(C), where k = 1.5 (for methyl H atoms) and k = 1.2 for other H atoms.The experimental details including the crystal data, data collection and refinement are summarized in Table 510.1107/S205698902000657X/ex2032sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698902000657X/ex2032Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902000657X/ex2032Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S205698902000657X/ex2032Isup4.cmlSupporting information file. DOI: 2004559CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Coronavirus disease-2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and represents a potentially fatal disease. COVID-19 is associated with a hypercoagulable state leading to increased incidence of venous thromboembolism. Arterial thrombosis has been reported, but the prevalence is not known. Herein, we report an unusual presentation of a 77-year-old male who presented with dyspnea and pain in left leg and was found to have acute limb ischemia. Our case adds to the limited literature regarding arterial thrombosis in COVID-19. At the end of 2019, a novel coronavirus was identified as the cause of a cluster of pneumonia cases in Wuhan, a city in the Hubei Province of China. On March 11, 2020, WHO declared coronavirus disease 2019 (COVID-19) as pandemic . We repoA 77-year-old male patient presented to emergency department with complaints of shortness of breath and pain, discoloration, and swelling of the left leg. Vital signs on presentation were heart rate 110 per minute, blood pressure 155/85 mmHg, oxygen saturation 96% on room air, and temperature 36 degree Celsius. The patient had bilateral crackles on lung exam and absent left dorsalis pedis and posterior tibial pulses and foot was swollen, discolored, and cold. Electrocardiogram showed sinus tachycardia 110 per minute.3 (reference: 4.5-11 K/mm3), platelets 534 K/mm3 (reference: 140-440 K/mm3), potassium 4.1 meq/L (reference: 3.5-5 meq/L), glucose 126 mg/dl (reference: 70-105 mg/dl), creatinine 0.61 mg/dl (reference: 0.6-1.30 mg/dl), blood urea nitrogen 8 mg/dl (reference: 7-23 mg/dl), lactic acid 1.1 mmol/L (reference: 0.5-2.2 mmol/L), troponin 0.136 ng/ml (reference: less than 0.03 ng/ml), D-dimer 2.77 (reference: less than 0.5), prothrombin time 14.8 seconds (reference: 12.2-14.9 seconds), international normalized ratio (INR) 1.2 (reference: less than 1), partial thromboplastin time 39.8 seconds (reference: 21.3-35.1 seconds), lactate dehydrogenase (LDH) 392 U/L (reference: 140-271 U/L), C-reactive protein (CRP) 301 mg/L (reference: less than 10 mg/L), ferritin 1,396 ng/mL (reference: 12-300 ng/ml), procalcitonin 0.60 ng/ml (reference: less than 2 ng/ml), aspartate transaminase 134 U/L (reference: 13-39 U/L), calcium 8.5 mg/dl (reference: 8.6-10.3 mg/dl), and albumin 2.7 mg/dl (reference: 3.5-5.0 mg/dl). Chest X ray showed bilateral hazy infiltrates. CT angiography of the abdomen and aorta with Iliofemoral runoff showed thrombus within the mid left superficial femoral artery and occluded left anterior tibial artery, left posterior tibial artery, and left peroneal artery with no flow to the foot , hematocrit 38.2% (reference: 36%-46%), white blood cells 41 K/mmCovid-19 was diagnosed on the basis of reverse transcription polymerase chain reaction (RT-PCR) testing. Echocardiogram showed an ejection fraction of 60%. The patient was placed on air-borne precautions and was started on ceftriaxone, azithromycin, hydroxychloroquine, and therapeutic anticoagulation with heparin. He underwent thrombectomy of left common femoral artery, profunda femoris, superficial femoral artery, popliteal artery, anterior tibial artery, posterior tibial artery, and peroneal artery.The clinical spectrum of COVID19 continues to evolve. The most common symptoms being reported are fever, myalgia, cough, and dyspnea, and less frequently headache, diarrhea, nausea, and vomiting . SeveritPatients with severe COVID-19 infection can develop a disseminated coagulopathy characterized by increases in procoagulant factors like fibrinogen and D-dimer leading to widespread microvascular thrombosis. This state has been termed thromboinflammation or COVID-19-associated coagulopathy . BellostAll hospitalized patients with COVID-19 should receive pharmacologic thromboprophylaxis with low molecular weight heparin (LMWH) or fondaparinux unless bleeding risk and full therapeutic-intensity anticoagulation in the appropriate clinical scenario . AnticoaWe report an unusual presentation of a 77-year-old male patient who presented with dyspnea and acute pain in left leg and was found to have ALI and underwent thrombectomy. Our case and review of literature reveals that health care providers should be aware of this unusual life-threatening manifestation of COVID 19 so that appropriate measures can be taken for the vascular emergency.


Right after the discovery of γδ T-cells in 1984, people started asking how γδ T-cells interact with other immune cells such as B-cells. Early reports showed that γδ T-cells are able to help B-cells to produce antibodies and to sustain the production of germinal centers. Interestingly, the presence of γδ T-cells seems to promote the generation of antibodies against “self” and less against challenging pathogens. More recently, these hypotheses were supported using γδ T-cell-deficient mouse strains, in different mouse models of systemic lupus erythematous, and after induction of epithelial cell damage. Together, these studies suggest that the link between γδ T-cells and the production of autoantibodies may be more relevant for the development of autoimmune diseases than generally acknowledged and thus targeting γδ T-cells could represent a new therapeutic strategy. In this review, we focus on what is known about the communication between γδ T-cells and B-cells, and we discuss the importance of this interaction in the context of autoimmunity. The adaptive immune system of vertebrates relies on three different lymphocyte lineages. B-, αβ T-, and γδ T-cells generate three different forms of clonal antigen receptors, namely the B-cell receptor (BCR), the αβ T-cell receptor (αβ TCR), and the γδ TCR, respectively.The γδ TCR was first cloned in 1984 and led to the discovery of γδ T-cells as an independent T lymphocyte lineage ,3. They γδ T-cells represent a bridge between the innate and the adaptive immune system: on one side, they can respond with relatively high clonal frequency and rapidly produce a variety of cytokines at the beginning of an immune response without the requirement of prior activation and clonal expansion. On the other side, they can behave like the adaptive arm of the immune response because of their pleiotropic effector functions and their clonal expansion after being primed via their TCR ,8.Trg and Trd loci. Failure to produce a functional γδ TCR drives T-cell progenitors with a functional TCR β-chain to enter the CD4+CD8+ double-positive (DP) stage where they rearrange their Tra loci and eventually display a functional αβ TCR. [trans-conditioning and depends on signals received through the lymphotoxin-β receptor, which serves pleiotropic functions including chemokine, cytokine or adhesion molecule expression, cell proliferation, and cell survival [+ and CD8+ αβ T-cell populations, in particular of cells with a memory phenotype [αβ and γδ T-cells derive from the same common lymphoid progenitor in the thymus. The development of γδ T-cells starts with the productive rearrangement of their  αβ TCR. . Not mucsurvival ,11,12. Mhenotype . Rather henotype .+ and Vγ4+ T-cells are mainly found in the secondary lymphoid organs. They are both very heterogeneous populations and can exert different functional effects in various mouse models of disease [+ T-cells are able to produce IFN-γ, IL-4, and IL-13, and a small subpopulation behave like NKT cells [+ T-cells can produce IL-17, TNF, IFN-γ, and IL-4, according to the studied model [+ cells are activated and produce IL-17 [In mice, Vγ1 disease . Vγ1+ T-KT cells ,16. Vγ4+ce IL-17 ,19.+ and Vδ2+ cells, whereas cells expressing other TCR δ chains are less frequent [+ T-cells are mainly localized at mucosal surfaces but can also be found in the blood, while Vδ2+ T-cells represent the main γδ T-cell subpopulation in the peripheral blood [+ T-cells, the Vδ2-chains pair with Vγ9-chains and this population can increase up to >50% of blood lymphocytes after certain bacterial or parasitic infections [Human γδ T-cells are subdivided based on the V-gene usage of the TCR δ chain. The most abundant and studied subpopulations in the human blood are Vδ1frequent . Vδ1+ T-al blood . In mostfections ,23.Current research on γδ T-cells focuses on discovering their TCR ligands in order to understand their functions and interactions within their tissue of residence. Still, an intriguing question is how γδ T-cells interact with other immune cells. In this review, we will focus primarily on the interaction between B and γδ T-cells and the contribution of γδ T-cells to humoral immunity and autoimmunity.It is well known that γδ T-cells have a strong impact on humoral immunity ,25,26,27+ T-cells due to the altered γδ T-cell compartment and its inability to produce IL-4. Interestingly, direct contact of MZ B-cells with γδ T-cells seems to be responsible for their decrease [Once B-cells express a rearranged BCR on the surface, they are negatively selected for self-antigen in the bone marrow during the first checkpoint of B-cell tolerance. Next, they leave the bone marrow and migrate via the bloodstream to the spleen. There, immature B-cells mainly differentiate through transitional stages into follicular and marginal zone (MZ) B-cells. By using γδ T-cell-deficient mouse strains, it has emerged that peripheral B-cells are influenced by the absence of γδ T-cells. Specifically, transitional, MZ B-, follicular B-, and the so called aged-associated B-cells (ABC)s were affected, indicating the capability of γδ T-cells to regulate precise stages of B-cell development . All thedecrease .After encountering their specific antigen, activated B-cells interact with their cognate T follicular helper (Tfh) cells, a subset of αβ T-cells which provide help to B-cells, resulting in their differentiation into extrafollicular short-lived plasma cells or their migration into the follicles to form germinal centers (GC)s ,30,31,32So, how could γδ T-cells potentially influence post bone marrow B-cell maturation and extrafollicular/GC B-cell differentiation into plasma cells? A number of interactions and mechanisms are conceivable : γδ T-ce+Vδ2+ T-cells isolated from peripheral blood expresses the CXC chemokine receptor type 5 (CXCR5) like Tfh cells, and, upon antigen stimulation, they are able to express the costimulatory molecules ICOS and CD40L, to produce cytokines such as IL-2, IL-4, and IL-10, and to help B-cells for the production of Abs [+Vδ2+ T-cell subset expresses CXCL13, CXCR5, and ICOS and provides help to B-cells in the presence of the phosphoantigen (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMB-PP) and the cytokine IL-21 [A subset of human Vγ9n of Abs . This Vγne IL-21 .+ T-cells can also upregulate CD40, CD86, and HLA-DR and promote the production and the release of IgM, but not IgG, IgA, or IgE by B-cells [More recently, it has been shown that Vδ3 B-cells . Togethe B-cells .−/−Tcrd mice immunized with ovalbumin presented far less Tfh cells compared to wild-type (WT) mice, suggesting a role for γδ T-cell in the development of Tfh. Indeed, they discovered, for the first time in mice, that a subpopulation of γδ T-cells expresses CXCR5, and, by releasing Wnt ligands, these cells are able to initiate the Tfh cell program in CD4+ cells. Interestingly, this γδ T-cell subpopulation can function as an APC to naïve T-cells [Besides their direct role in helping B-cells, γδ T-cells can also regulate Tfh cells. Rezende and colleagues showed that  T-cells .+Vδ2+ T-cells display the main characteristics of a professional APC, they efficiently process and display the antigens on MHCII molecules, and provided co-stimulatory signals for strong induction of naïve CD4+ T-cell proliferation and differentiation [In humans, phosphoantigen-activated Vγ9ntiation .+ T, γδ T, NKT, B-cells, basophils, eosinophils, mast cells, and also by type-2 innate lymphoid cells. In mice, IL-4 induces the differentiation of naïve CD4+ T-cells into Th2 cells, drives the Ig class switch to IgG1 and IgE in B-cells, and induces alternative macrophage activation [IL-4 is a typical signature cytokine of the type II inflammatory response triggered during parasitic infections and allergy. IL-4 can be produced by CD4tivation . IL-4 cativation ,46.+ and Vγ6+ T-cell populations, which also presented increased levels of IL-4 in the serum [Vγ4−/−/6−/− mice were then crossed with IL-4 KO mice to remove any possible effect of IL-4, whose production is enhanced by Vγ1+ cells that developed in the absence of Vγ4+. The lack of IL-4 in the double mutant mice abrogated the phenotype of Vγ4−/−/6−/− mice, indicating an important role of γδ T-cells in the regulation of IL-4 and Ig production [Early experiments conducted in mice that congenitally lack αβ T-cells showed that their B-cells could still expand and secrete Abs of the subclasses IgG1 and IgE, suggesting for the first time a role for IL-4 producing γδ T-cells in helping B-cells . Similarhe serum . These Voduction .+ intraepithelial lymphocytes (IEL)s could trigger the production of IgE, which confer protection in the exposed tissue. Interestingly, this endogenous IgE response was autoreactive and supported by IL-4-producing CD4+ αβ T-cells. In addition, sequencing of IgE antibodies in WT and Tcrd−/− mice revealed that Tcrd−/− mice had a defective clonal expansion of IgE+ B-cells and a less diverse B-cell repertoire compared to WT mice. Intriguingly, when using another chemical agent, TPA (12-O-tetradecanoylphorbol-13-acetate), which does not induce DNA damage but triggers skin inflammation, the induced IgE antibodies were not autoreactive. In summary, γδ TCR+ IELs could shape a distinct IgE repertoire that was dependent on the presence of DNA damage in epithelial cells, indicating a unique role of γδ TCR+ IELs in initiating and regulating IgE production and repertoire [More recently, Crawford and colleagues studied a model of epithelial damage in the skin induced by using the DNA-damaging xenobiotic and carcinogen DMBA  . In thispertoire .+Vδ2+ T-cells as they are able to activate specific γδ TCRs [+ T-cells also seem to be stimulated by activated B-cells isolated from peripheral blood [+ T-cells [So far, we have focused on the influence of γδ T-cells on B-cells, but what about vice versa? Already in 1989, Sperling and Wortis found that a syngenic B-cell lymphoma induced proliferation of murine γδ T-cells after co-culture . Similar γδ TCRs ,51,52. Ial blood ,54. More T-cells . Therefo T-cells .+Vδ1+ T-cells [+Vδ2+ T-cells and Vγ4+ T-cells) [Finally, B-cells may interact with γδ T-cells via butyrophilins (BTN)s. BTNs are members of the B7 family of costimulatory receptors, which include B7.1 (CD80) and B7.2 (CD86), and have also been recognized as potentially important immune modulators . They ar T-cells . More reT-cells) ,60,61,62T-cells) .Tcrd−/− mice constitutively depleted in the δ chain [Tcrd-GDL mice where the γδ T-cells can be conditionally depleted through the injection of diphtheria toxin (DTx) as the mice express a human DTx receptor. Therefore, this model will allow the depletion of γδ T-cells without the compensatory effects of αβ T-cells and compromising other αβ T-cells [In order to better study the effect of B-cells on γδ T-cells, it is important to find the right experimental approach. Previously, a common way to investigate the function of γδ T-cells was the depletion of γδ T-cells via the use of monoclonal antibodies anti-γδ TCR which block and internalize the γδ TCR ,65. The  δ chain  can be u δ chain ,69. This δ chain . A new w T-cells .The GC reaction plays an important role in humoral immunity, as B-cells with the highest affinity to GC-contained antigens are selected to become plasma cells that produce high-affinity antibodies or memory B-cells ,73,74. GEarly studies have shown that γδ T-cells can be found inside GCs ,77. FurtTcra−/− and Tcrb−/− mice  upon infection [+ and Vγ6+ γδ T-cells showed a decrease in total Ig level in sera. In contrast, mice carrying only Vγ1+ γδ T-cells showed an increase of total Ig level and displayed spontaneous GCs in the spleen [Tcrd−/− mice immunized with CFA and OVA showed reduced frequencies of GC B-cells due to the missing help of the Tfh cells [Hayday and collaborators showed in the 1990s that GCs can be formed in nfection ,80. Subsnfection . Mice the spleen . As mentfh cells .+Vδ2+ T-cells. These γδ T-cells stimulated with phosphoantigens could provide B-cell help through cell–cell contact. Within 36 h after stimulation, CD40L, OX40, and ICOS were highly upregulated, and, in co-culture experiments, they acted as strong B-cell help for production of IgM-, IgG-, and IgA-like Tfh cells [+Vδ2+ T-cells activated with the phosphoantigen HMB-PP and in presence of IL-21, which is mainly produced by Tfh in the GC, showed production of CXCL13. This chemokine is normally produced by FDCs in the light zone of the GC in order to guide B-cells into the light zone [Further evidence that γδ T-cells play a role in providing help to B-cells was found in experiments involving human blood Vγ9fh cells . In the ght zone ,82,83. αght zone .Together, these findings suggest an involvement of γδ T-cells in B-cell class-switching (extra-follicular or within the GC) and in supporting GC reactions at different stages . AdditioAfter reviewing the possible ways of γδ-B-cell interaction, the next question is why γδ T–B-cell interaction would be important. Clearly, there is a positive association between γδ T-cells and autoimmunity ,86. HoweTcra−/− mice is that their serum antibodies are more autoreactive than those of WT control mice. In particular, they are positive for anti-nuclear antibodies (ANA)s, anti-double stranded (ds)-DNA and small nuclear ribonucleoproteins which are typical autoantibodies that are characteristic of systemic lupus erythematosus (SLE) [+ T-cells and, mainly, αβ T-cells (regulated by the γδ T-cells) for producing large amounts of IL-4 that activate B-cell maturation and can overrun tolerance mechanisms [−/−Tcrb mice were able to generate self-reactive antibodies after parasitic infection, in particular towards DNA instead of antibodies specific for the pathogen, thereby supporting the idea that γδ T-cells are more important for autoantibody production rather than mounting a pathogen-specific immune reaction [What it is known so far from us (SLE) ,87. Simichanisms ,15,47. Ichanisms ,89,90. Ireaction .Tcrd−/− mice at steady state and after induction of a murine model of SLE. Tcrd−/− mice showed decreased autoantibody production at steady state and upon induction of SLE [Recently, an autoantibody microarray was performed on serum from WT and n of SLE . Possibln of SLE  or that n of SLE . At thisThus, γδ T-cells seem to play an important role in the regulation of human autoimmune diseases such as inflammatory bowel disease and experimental autoimmune encephalomyelitis . Moreoveγδ T-cells seem to have the potential to regulate B-cell maturation during their development in the periphery (spleen) and during an immune response (in GC). Whether such an influence of γδ T-cells is mediated via soluble mediators important for B-cells (such as IL-4), or rather by the presentation of autoantigens, remains to be determined . At this


In the extended structure, two molecules arranged in a near coplanar position relative to the triazole ring planes are interconnected by N—H⋯N and C—H⋯N hydrogen bonds into a homodimer. The dimers are linked by C—H⋯O interactions into ribbons.The title compound was obtained  19H17ClN4O2, was obtained via a two-step synthesis involving the enol-mediated click Dimroth reaction of 4-azidoanisole with methyl 3-cyclopropyl-3-oxopropanoate leading to the 5-cyclopropyl-1-(4-methoxyphenyl)-1H-1,2,3-triazole-4-carboxylic acid and subsequent acid amidation with 4-chloroaniline by 1,1′-carbonyldiimidazole (CDI). It crystallizes in space group P21/n, with one molecule in the asymmetric unit. In the extended structure, two molecules arranged in a near coplanar fashion relative to the triazole ring planes are interconnected by N—H⋯N and C—H⋯N hydrogen bonds into a homodimer. The formation of dimers is a consequence of the above interaction and the edge-to-face stacking of aromatic rings, which are turned by 58.0 (3)° relative to each other. The dimers are linked by C—H⋯O interactions into ribbons. DFT calculations demonstrate that the frontier molecular orbitals are well separated in energy and the HOMO is largely localized on the 4-chlorophenyl amide motif while the LUMO is associated with aryltriazole grouping. A Hirshfeld surface analysis was performed to further analyse the intermolecular interactions.The title compound, C Atom O1 of the amide group accepts both an intramolecular C—H⋯O link (with the 4-chlorophenyl and cyclopropyl H atoms) and an intermolecular C2—H2⋯O1 interaction with the 4-methoxyphenyl H atom. The last of these links neighbouring dimers into hydrogen-bonded ribbons parallel to the [010] direction  using CrystalExplorer  atoms of neighbouring molecules) can be seen in the Hirshfeld surface plot as red areas -1-(4-chlorophenyl)-N-(−1-hydroxy-3-phenylpropan-2-yl)-5-methyl-1H-1,2,3-triazole-4-carboxamide (I)et al., 2013N-[methyl]-1H-1,2,3-triazole-4-carboxamide (II) morpholino)methanone (III)  P21/c in (II)], while compounds (III) and (IV) crystallize in the triclinic space group P12H13N5O)(NO3)]·0.5H2O and [Cu(C12H13N5O)(CF3COO)]  were obtained by electrochemical synthesis  atoms, infinite chains containing [CuC4NO] seven-membered rings.Compounds (I)H-1,2,3-triazole-4-carboxylic acid  was added to a solution of 1,1′-carbonyldiimidazole  in dry acetonitrile (25 ml) and the mixture was kept for 30 min at 323 K. Then 4-chloroaniline 2  was added, and the mixture was heated at 343 K for 1 h. After cooling to room temperature, water (30 ml) was added. The precipitate was filtered off, washed with water on a filter, recrystallized from ethanol solution, and dried in air to give the title compound as colourless prismatic crystals, m.p. 422–423 K; 1H NMR  δ 10.56 , 7.89 , 7.58 , 7.39 , 7.16 , 3.86 , 2.10–1.99 , 0.95–0.80 ; 13C NMR  δ 160.62 (C=O or CAr—O), 159.60 (C=O or CAr—O), 142.26 (CTriazole-4), 138.87 (CTriazole-5), 138.21 (CClAr-1), 129.08 (CAr-1), 128.91 , 127.77 , 127.70 (CClAr-4), 122.25 , 115.00 , 56.06 (MeO), 8.09 (2 × CH2cPr), 5.75 (CHcPr); MS m/z = 369 (M++1); Analysis calculated for C19H17ClN4O2 (Mr = 368.82), (%): C 61.88, H 4.65, N 15.19; found (%): C 61.91, H 4.74, N 15.21.The title compound was synthesized from 5-cyclopropyl-1-(4-methoxyphenyl)-1Uiso(H) = 1.2Ueq(carrier) or 1.5Ueq(C-methyl carrier) was applied in all cases.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989020005848/hb7901sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989020005848/hb7901Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020005848/hb7901Isup3.molSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020005848/hb7901Isup4.cmlSupporting information file. DOI: 1999643CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Engagement of integrin αIIbβ3 promotes platelet–platelet interaction and stimulates outside-in signaling that amplifies activation. Protein kinase Cδ (PKCδ) is known to play an important role in platelet activation, but its role in outside-in signaling has not been established. In the present study, we determined the role of PKCδ and its signaling pathways in integrin αIIbβ3-mediated outside-in signaling in platelets using PKCδ-deficient platelets. Platelet spreading to immobilized fibrinogen resulted in PKCδ phosphorylation, suggesting that αIIbβ3 activation caused PKCδ activation. αIIbβ3-mediated phosphorylation of Akt was significantly inhibited in PKCδ -/- platelets, indicating a role of PKCδ in outside-in signaling. αIIbβ3-mediated PKCδ phosphorylation was inhibited by proline-rich tyrosine kinase 2 (Pyk2) selective inhibitor, suggesting that Pyk2 contributes to the regulation of PKCδ phosphorylation in outside-in signaling. Additionally, Src-family kinase inhibitor PP2 inhibited integrin-mediated Pyk2 and PKCδ phosphorylation. Lastly, platelet spreading was inhibited in PKCδ -/- platelets compared to the wild-type (WT) platelets, and clot retraction from PKCδ -/- platelets was markedly delayed, indicating that PKCδ is involved in the regulation of αIIbβ3-dependent interactivities with cytoskeleton elements. Together, these results provide evidence that PKCδ plays an important role in outside-in signaling, which is regulated by Pyk2 in platelets. Integrins play a key role in regulating platelet functions, including platelet adhesion, spreading, clot retraction, and platelet pro-coagulant activity. Engagement of αIIbβ3 promotes platelet–platelet interaction  and genePKC is a serine/threonine-related protein kinase and has multiple isoforms classified to three subfamilies . The con2 (TxA2) generation, and thrombin- and collagen-induced PKCδ phosphorylation is regulated by αIIbβ3 outside-in signaling, raising the potential role of PKCδ in αIIbβ3-mediated signaling [A previous study implicated PKC isoforms in the regulation of integrin properties in many cell types , but theignaling . Thus, oIn this study, we found that PKCδ is phosphorylated by integrin αIIbβ3-mediated outside-in signaling in platelets. We further found that integrin αIIbβ3-mediated PKCδ phosphorylation is regulated by Src and Pyk2, and αIIbβ3-mediated phosphorylation of Akt is regulated by PKCδ. Furthermore, PKCδ contributes to regulation of platelet spreading and clot retraction. Therefore, we concluded that PKCδ plays an important role in integrin αIIbβ3-mediated outside-in signaling in platelets.To investigate the signaling mechanism of PKCδ activation in outside-in signaling, we first evaluated the role of integrin αIIbβ3 in PKCδ activation. As shown in Src-family kinases (SFKs) are the major tyrosine kinases downstream of integrin αIIbβ3-mediated signaling in platelets and also mediate PKCδ phosphorylation downstream of PARs in platelets . To deteTo determine the role of PKCδ in integrin-mediated outside-in signaling, we tested the effect of PKCδ on αIIbβ3-mediated Pyk2 and Akt phosphorylation using PKCδ-deficient platelets. As shown in To determine the functional role of PKCδ in outside-in signaling in platelets, we examined the effect of PKCδ on platelet spreading on immobilized fibrinogen and clot retraction using PKCδ-deficient platelets. As shown in PKCδ is abundantly expressed in platelets and is activated by various agonists including thrombin, collagen, ADP, and von Willebrand factor (vWF) . Regulatq-mediated pathway [q-signaling. PKCδ is phosphorylated on Thr505 and Tyr565 ,28. It i pathway . Thus, tIntegrin αIIbβ3-mediated outside-in signaling involves a number of known intracellular effectors, including SFKs, Pyk2, and Akt ,26,30. SIntegrin αIIbβ3-mediated outside-in signaling was found to be critically important in stable platelet adhesion, spreading, and clot retraction . We obseIn conclusion, we demonstrated that PKCδ is activated downstream of αIIbβ3-mediated outside-in signaling in platelets. In addition, PKCδ is regulated by Src and Pyk2 in platelets. Finally, PKCδ is an important regulator of integrin αIIbβ3 outside-in signaling and contributes a major role in platelet adhesion, spreading, and clot retraction.2-MeSADP, thrombin, apyrase (type V), prostaglandin E1 (PGE1), bovine serum albumin (fraction V), fibrinogen, sodium citrate, and acetylsalicylic acid were purchased from Sigma . AYPGKF was custom synthesized by Invitrogen . Rhodamine phalloidin was from Invitrogen . Anti-phospho-Pyk2 (Tyr402), anti-phospho-Akt (Ser473), anti-phospho-PKCδ (Tyr311), and anti-β-actin antibodies were purchased from Cell Signaling Technology . PP2, PP3, and GR144053 were from Enzo Life Sciences . TAT-Pyk2-CT and TAT-GFP controls were generously provided by Xiangdong Zhu, University of Chicago . Secondary antibody was from Santa Cruz Biotechnology . All other reagents were reagent grade. All animal experiments in this study were performed in line with the approval obtained from the Chungbuk National University Animal Ethics Committee . PKCδ-deficient mice were a generous gift from Dr. Keiko Nakayama . Wild-type littermates were used as controls.g for 10 min and 400× g for 10 min at room temperature (RT) to obtain the platelet-rich plasma (PRP) and platelet pellet, respectively. Washed platelets were prepared by re-suspending the platelet pellet in Tyrode’s buffer and adjusted to 2 × 108 cells/mL.Murine platelets were prepared by collecting whole blood from an equal number of both male and female mice as described previously . BrieflyPhosphorylation events were measured from AYPGKF- and 2-MeSADP-stimulated platelets or fibrinogen-adherent platelets as described previously . Briefly2 and 2 mg/mL fibrinogen. Clot retraction was initiated by adding 0.1 U/mL of thrombin, allowed to proceed at 37 °C, and then photographed at indicated time points.Washed platelets were plated on fibrinogen-coated coverslips and platelet spreading was observed as previously described . Brieflyt-test using GraphPad Prism software (version 3.0) . The values were shown as mean ± SE. The significance of difference between data was analyzed by Student’s


Integrity between neighboring molecules in the solid state is maintained by an array of C—H⋯N hydrogen bonds and edge-to-face C—H⋯π interactions.The single-crystal X-ray structure of  trans-bis(pyridin-3-yl)ethylene , C12H10N2, as determined by single-crystal X-ray diffraction is reported. The molecule self-assembles into two dimensional arrays by a combination of C—H⋯N hydrogen bonds and edge-to-face C—H⋯π interactions that stack in a herringbone arrangement perpendicular to the crystallographic c-axis. The supramolecular forces that direct the packing of 3,3′-bpe as well as its packing assembly within the crystal are also compared to those observed within the structures of the other symmetrical isomers trans-1,2-bis(n-pyridyl)ethylene .The molecular structure of  Unlike cinnamic acid, which crystallizes in such a way that the olefins are rendered photoactive (olefins within 4.2 Å of one another), the native crystalline forms of bis(pyridyl)ethylenes are photostable . To achieve photoreactivity of these olefins, it often becomes necessary to use a ‘molecular template’ that can interact with the olefin-containing bipyridine via supramolecular interactions such as hydrogen bonding, halogen bonding, argento- and aurophilic interactions, and dative N→B interactions. Analyses of the crystal structures of symmetric bis(pyridyl)ethylenes derivatives such as the trans-bis(n-pyridyl)ethylenes series of isomers  is necessary to understand the forces that govern their crystallization, why they are photostable, and why use templates to achieve photoreactivity ethylenes have arisen as somewhat of a natural extension of cinnamic acid as a series of molecules capable of undergoing [2+2] photodimerization in the solid state to generate cyclobutanes. Foundational work by Schmidt and coworkers on 3,3′-bpe crystallizes in the centrosymmetric monoclinic space group P21/n  3.58 Å; Θ(C6—H6⋯pyr) 131.8°] forces between pyridyl rings  3.59 Å; Θ(C4—H4⋯N1) 139.5°] hydrogen bonds  series of symmetric alkenes, all three adopt nearly planar conformations  structures  furnished 3,3′-bpe as yellow crystals: 222.3 mg (23%). A portion of 3,3′-bpe was dissolved in CHCl3 and allowed to slowly evaporate at room temperature. Single crystals in the form of colorless plates suitable for single crystal X-ray diffraction formed within seven days.The alkene 3,3′-bpe are summarized in Table 2Crystal data, data collection and structure refinement details for 10.1107/S2056989020015303/dj2017sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020015303/dj2017Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020015303/dj2017Isup3.cmlSupporting information file. DOI: 1985201CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Futhermore, the crystal structures of (1) and (2) were refined using independent atom model (IAM) and Hirshfeld atom refinement (HAR) approachesThe crystal structures of trismethylsilane ( 13H18O3Si (1) and C18H20O3Si (2), represent functionalizable dihydrofuranylsilanes, which permit substitution by a variety of nucleophiles. The crystal structures of 1 and 2 display weak intermolecular C—H⋯O hydrogen-bonding interactions , leading to a two-dimensional supramolecular network for 1 and a one-dimensional supramolecular network for 2. The crystal structures of 1 and 2 were refined both on the basis of the independent atom model (IAM) and the Hirshfeld atom refinement (HAR) approach, and the results are comparatively discussed.The title compounds, C An approach that takes this into consideration is Hirshfeld atom refinement (HAR), which uses aspherical atomic scattering factors calculated from tailor-made 1 and 2, the minimum and maximum values of residual electron density are significantly lower than those of IAM results . In all cases, the residual densities do not indicate any disorder. For compound 1, the residual electron density on the basis of the HAR refinement is close to O1 and H8A and for 2 is near C15 and H3B. Another aim of the Hirshfeld atom refinement was the accurate localization of hydrogen atoms. From a comparison of the C—H bond lengths of the methine groups using IAM and HAR approaches, it can be clearly observed that the C—H bonds of the HAR model are significantly longer than those of the AIM model  structure refinements of compounds with dihydrofuranyl rings performed by our group, we observed slight disorders of the oxygen atom and the methine atom of the dihydrofuranyl ring. Therefore, results of HARs for such compounds are interesting in order to draw conclusions about the residual electron densities to exclude and/or model disorder. For l Table 5. Woińska al. 2016 have alrR1 value of 0.023 was observed for compound 1, compared to the refinement using IAM with an R1 value of 0.035 (compound 2: R1 for HAR = 0.024 versus IAM = 0.037).When using HAR, an improved 1, the molecules are linked by a number of C—H⋯O hydrogen bonds, forming a network along the [012] direction  is a colourless crystalline solid at room temperature:Trismethylsilane : δ = 0.65 , 2.25 , 4.06 , 5.59  ppm.1H}13C NMR : δ = −5.7 , 31.4 , 70.9 , 115.6 , 157.5  ppm.{1H}29Si NMR : −36.65  ppm.{tR = 5.40 min [353 K (1 min) – 40 K min−1 – 543 K (5.5 min)]; m/z (%): 250 (100) [M+], 207 (4) [(M − C2H3O)+], 121 (56) [ [(SiDHF)+].GC/EI–MS 2) is a colourless crystalline solid at room temperature:Trisphenylsilane : δ = 2.25 , 4.07 , 5.72 , 7.18–7.27 , 8.11–8.14  ppm.1H}13C NMR : δ = 31.4 , 71.1 ; 117.8 ; 128.4 ; 130.8 ; 134.3 ; 136.3 ; 156.4  ppm.{1H}29Si NMR : −41.74  ppm.{tR = 6.88 min [353 K (1 min) – 40 K min−1 – 543 K(5.5 min)]; m/z (%): 312 (100) [M+], 255 (21) [(M − C3H5O)+], 105 (53) [ [Ph+], 69 (6) [DHF+].GC/EI–MS SHELXL  = 1.2Ueq(C) for CH2 and CH hydrogen atoms and Uiso(H) = 1.5Ueq(C) for CH3 hydrogen atoms. Hydrogen atoms H6, H8A,B and H11A,B for compound 1 and H2 and H16 for compound 2 were refined independently.Crystal data, data collection and structure refinement details are summarized in Table 6OLEX2  1, 1HAR, 2, 2HAR. DOI: 10.1107/S2056989020011470/wm55611sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989020011470/wm55611HARsup4.hklStructure factors: contains datablock(s) 1HAR. DOI: 10.1107/S2056989020011470/wm55612sup3.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S2056989020011470/wm55612HARsup5.hklStructure factors: contains datablock(s) 2HAR. DOI: 2024677, 2024676, 2024675, 2024674CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Their structures were determined by MS, NMR, and X-ray spectroscopic techniques. Among the methylated flavonols, 1, 4–7, and 9 were isolated for the first time from the rhizomes, while 1, 4, and 5 were isolated from the fruit. Compounds 2, 3, 7, 8, and 10 were reported for the first time from the species. Three main methylated flavonols 1, 4, and 5 were quantitatively analyzed in the rhizomes of A. koenigii by RP-HPLC-DAD; their contents were determined to be 1.81% (1), 1.38% (4), and 1.76% (5). The antimicrobial assay against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, Aspergillus niger, Fusarium oxysporum, Candida albicans, and Saccharomyces cerevisiae and antioxidant DPPH scavenging test were performed for the isolated methylated flavonols.Methylated flavonols form a special group with modulating biological activities in comparison with kaempferol and quercetin. The present study isolated ten compounds including two kaempferol methyl ethers: 5-hydroxy-3,7,4′-trimethoxyflavone ( Amomum Roxb. is a large genus with about 250 species and belongs to Zingiberaceae. In recent years, 21 Amomum species have been recorded in Vietnam [Amomum species from Vietnam has been studied [A. koenigii has been used as an aromatic stomachic in China, India, and Thailand [A. koenigii in China describes the isolation of eicosenones and methylated flavonols from the fruit of A. koenigii [A. koenigii from Vietnam  was used as the NMR internal standard. Diaion HP-20  and silica gel  of 40–63 and 15–40 μm were used for column chromatography (CC). Thin-layer chromatography (TLC) was performed on Merck precoated aluminum silica gel 60 F254 plates. Tryptone soya agar  and Sabouraud's dextrose agar  were used for the antimicrobial activity assay. Reagents and standard compounds used in the antioxidant DPPH scavenging assay were purchased from Sigma-Aldrich (USA).ESI-MS spectra were measured on a Thermo Fisher Scientific LTQ Orbitrap XL mass spectrometer in CHA. koenigii were collected in July 2016 from Chu Yang Sin National Park, Hoa Son Commune, Krong Bong District, Dak Lak Province, Vietnam (coordinates: from 12°14′16″ north to 13°30′58″ north and from 108°17′47″ west to 108°34′48″ west). The plant material was identified by Dr. Quoc Binh Nguyen, Vietnam National Museum of Nature, Vietnam Academy of Science and Technology, Hanoi, Vietnam. A voucher sample (No. AK-7-17) was deposited in the same museum.The fresh fruit (3 kg) and rhizomes (16 kg) of n-hexane and dichloromethane. Evaporation of solvents under reduced pressure gave n-hexane- and dichloromethane-soluble fractions. The n-hexane- and dichloromethane-soluble fractions were combined based on their TLC similarities, and 20.2 g of the combined fraction was separated by CC over silica gel, eluting with n-hexane-acetone 19 : 1, 15 : 1, 9 : 1, 5 : 1, and 1 : 1 (v : v) to give 11 fractions (Fr. 1–Fr. 11). Fr. 2 (0.83 g) was purified by CC over silica gel, eluting with n-hexane-EtOAc 49 : 1, 29 : 1, 19 : 1, 9 : 1, and 6 : 1 to give 1 (112 mg) and 2 (20.1 mg). Frs. 4 and 5 were combined (0.77 g) and separated by CC over silica gel, eluting with n-hexane-acetone 20 : 1, 15 : 1, 12 : 1, 9 : 1, and 3 : 1 to yield 3 (5 mg). Fr. 7 (1.73 g) was separated by CC over silica gel, eluting with n-hexane-acetone 20 : 1, 15 : 1, 9 : 1, 6 : 1, 3 : 1, and 1 : 1 to yield 1 (36.9 mg), 4 (8.2 mg), and 5 (25.4 mg). Frs. 8 and 9 were combined (3.15 g) and separated by CC over RP-18, eluting with 70%, 80%, and 90% MeOH-H2O and MeOH. The fraction eluting with MeOH was separated by CC over silica gel, eluting with n-hexane-acetone 15 : 1, 12 : 1, 9 : 1, 6 : 1, and 3 : 1 to give 1 (5 mg). Fr. 10 (2.41 g) was separated by CC over RP-18, eluting with 70%, 80%, and 90% MeOH-H2O and MeOH. The fraction eluting with 70% MeOH-H2O (1.56 g) was separated by repeated CC over silica gel, eluting with n-hexane-acetone 25 : 1, 20 : 1, 12 : 1, 9 : 1, 6 : 1, and 3 : 1 to give 4 (2 mg) and 5 (20 mg). Fractions eluting with 80% and 90% MeOH-H2O were combined (0.54 g) and separated by CC over silica gel, eluting with n-hexane-EtOAc 12 : 1, 9 : 1, 6 : 1, 3 : 1, 1 : 1, and 1 : 2 to give 4 (5 mg), 6 (7.3 mg), 7 (30.2 mg), 8 (3 mg), and 9 (6 mg).The rhizomes were air-dried and then oven-dried at 40–50°C. The dried material was ground into powder. The powder (2.7 kg) was macerated with MeOH at room temperature three times, each time for 7 days. The extracts were filtered and concentrated under reduced pressure to give the MeOH extract. The MeOH extract was suspended in water, and the water phase was extracted with A. koenigii was extracted with methanol at room temperature five times, each for five days. The methanol extracts were combined and evaporated under reduced pressure. The residue was suspended in water and extracted with n-hexane and dichloromethane, and the organic phases were evaporated under reduced pressure to give n-hexane- and dichloromethane-soluble fractions. Half of the n-hexane-soluble fraction (12.4 g) was separated by CC over silica gel, eluting with CH2Cl2-MeOH 29 : 1, 19 : 1, 9 : 1, 6 : 1, 3 : 1, and 1 : 1 to give two fractions (Frs. 1 and 2). Fr. 1 (4.7 g) was separated by CC over silica gel, eluting with n-hexane-EtOAc 29 : 1, 19 : 1, 9 : 1, 6 : 1, 3 : 1, and 1 : 1 to give 1 (15.0 mg), 4 (6.0 mg), 5 (8.0 mg), and 8 (5 mg). The water-soluble fraction was passed through a column packed with Diaion HP-20, eluting with 20% and 60% MeOH-H2O and MeOH. The fraction eluting with 20% MeOH-H2O (0.1 g) was separated by CC over silica gel, eluting with CH2Cl2-EtOAc 15 : 1, 12 : 1, 9 : 1, 6 : 1, 3 : 1, and 1 : 1 to give 8 (10 mg) and 10 (46 mg). The fraction eluting with 60% MeOH-H2O (0.2 g) was separated by CC over silica gel, eluting with CH2Cl2-MeOH 30 : 1, 25 : 1, 15 : 1, 12 : 1, 9 : 1, 6 : 1, 3 : 1, and 1 : 1 to give 10 (34 mg).The fruit was washed, air-dried to remove surface water, and then crushed. The crushed fruit (3 kg) of µm particle analytical RP-18 column (4.6 mm × 250 mm). The mobile phase was acetonitrile (HPLC grade)-deionized H2O gradient: 56% acetonitrile (0–5 min) and 100% acetonitrile (5.1–15.5 min). The flow rate was 1 mL/min. The injection volume was 20 μL. The column temperature was 30°C. The HPLC conditions were optimized for the baseline, precision, accuracy, repeatability, LOD, and LOQ at the corresponding signal-to-noise ratios 3 and 10  and the peak area (y-axis) was constructed. The calibration standards were prepared by serial dilution of 1000 ppm concentration stock solutions of 1, 4, and 5 (HPLC purity ≥ 95%) . The procedures were done using the following tools: absorption correction: multi-scan, SADABS 2016/2 ; data collection: APEX3 ; cell refinement: SAINT ; data reduction: SAINT ; programs used to solve and refine structure: SHELXTL ; molecular graphics: SHELXTL ; and software used to prepare the material for publication: SHELXTL . Reflections were collected at 100 K.X-ray diffraction data were collected on a Bruker APEX-II CCD diffractometer with Mo K8 colony forming units (CFU/mL) (0.5 McFarland standard). The samples were dissolved in dimethylsulfoxide (DMSO) by vortexing and filtered through a 0.02 μm microfilter to produce stock solutions. The wells were filled with 50 μL stock solutions in DMSO and 50 μL organism suspension. The positive controls were prepared in DMSO: ampicillin (50 mM), tetracycline (10 mM), and nystatin (0.04 mM). MIC values of the positive controls were as follows: ampicillin 21.83 μg/mL, tetracycline 6.87 μg/mL, and nystatin 1.44 μg/mL (for yeasts) and 2.89 μg/ml (for fungi). The negative control was DMSO. The wells were allowed to diffuse for 24-hour incubation at 37°C and 48-hour incubation at 30°C for bacteria and fungi, respectively. The results were positive when no growth was observed (CFU < 5). MIC was determined by using serial dilution of the stock solutions. MIC is defined as the lowest concentration of the compounds that showed no growth compared with growth in control (DMSO) wells. All determinations were performed in triplicate.The antimicrobial assay was performed in 96-well plates, and minimum inhibitory concentration (MIC) was determined as described by Vanden Berghe and Vlietinck . The bac50, and the value was determined using TableCurve AISN Software (Jandel Scientific) based on the calculated values of the DPPH scavenging activity (%) of the sample. DPPH scavenging activity (%) was calculated with the following formula: scavenging capacity (SC) (%) = 100 − (A1 − A0)/Acontrol × 100, where A0 is the absorbance of the blank (sample in DMSO), Acontrol is the absorbance of the negative control, while A1 is the absorbance in the presence of the sample [Approximately, 3 mL of 0.1 mM solution of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical in EtOH 96% was each added into six series of prepared concentrations of sample solutions (1 mL in DMSO). The test was performed in triplicate. The solution was mixed vigorously and left to stand at room temperature for 30 min in the dark after which its absorbance was measured spectrophotometrically at 515 nm using an ultraviolet spectrophotometer. DMSO was used as the blank and negative control (1 mL DMSO mixed with 3 mL DPPH), and quercetin was used as the standard. The concentration of the sample required to inhibit 50% of DPPH free radicals is denoted as SCe sample .1) [β-sitosterol (2) [3) [4) [5) [6) [7), p-hydroquinone (8) [9) [10) [A. koenigii  , β-sitoserol (2) , 5-hydro (2) [3) , retusin [3) [4) , 3,5-dih [4) [5) , 3-hydronone (8) , 3,5,7,3 (8) [9) , and van[9) [10) , 17 werekoenigii . The met1  and 4  were prepared by recrystallization. We confirmed the locations of the methyl substituents  in 1 and 4 by X-ray crystallographic analysis. The unit cell of 1 was determined to be the triclinic crystal system, P-1 space group with Z = 2 and molecular formula C18H16O6    showed three main peaks in the HPLC chromatogram of the rhizome MeOH extract of A. koenigii from Vietnam. The retention times of 1, 4, and 5 were 12.6, 11.9, and 10.3 min, respectively. According to the HPLC quantification, the flavonoids were present in the rhizomes of A. koenigii in dry quantities of 1.81% (1), 1.38% (4), and 1.76% (5). The content of the major compound 4 (retusin) was almost 0.5% in the seeds [Previously, compounds of China . A typic1 and 4–7 were tested for their antibacterial and antifungal activity as described by Vanden Berghe and Vlietinck [Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 25923), Bacillus subtilis (ATCC 27212), Staphylococcus aureus (ATCC 12222), Aspergillus niger (439), Fusarium oxysporum (M42), Candida albicans (ATCC 7754), and Saccharomyces cerevisiae (SH 20). The samples were tested for their antimicrobial activity, and the active samples underwent serial 2-fold dilutions to determine MIC values. The MIC is defined as the lowest concentration of compounds that inhibits any visible growth of microorganisms after incubation. Compounds were tested at the concentrations of 100 μg/mL  and 50 μg/mL (6 and 7); only 4 showed potent activity against A. niger with an MIC value of 100 μg/mL (S. aureus (MIC 100 μg/mL), P. aeruginosa (MIC 100 μg/mL) [E. coli (MIC50 35.76 μg/mL) [4, 5, and 7 decreased the antibacterial activity, and the presence of the hydroxyl groups at C-3′ and C-4′ in the catechol moiety of the flavonoid B ring was important for the activity, while the free hydroxyl groups at C-3, C-5, and C-4′ were not critical. Kaempferol is much less active than quercetin, showing MIC values against S. aureus and B. cereus of 500 μg/mL, but it is strongly active against E. coli (MIC50 25 μg/mL) [6 showing that the free hydroxyl group at C-3 was not critical for strong antibacterial activity.Five methylated flavonols 00 μg/mL . Quercet0 μg/mL) , and E. 6 μg/mL) . In accoisiae SH . The sams ATCC 272, Staphy4, 5, and 7 was evaluated based on the scavenging activity test against DPPH. Compounds 4, 5, and 7 were selected because quercetin with its catechol  structure in the flavonoid B ring is a much stronger antioxidant than kaempferol [λ = 515 nm. The tests were triplicated, and the results were averaged (p < 0.05). As a result, compounds 4  and 5  showed a lower scavenging activity in comparison with that of quercetin   in comparison with those of 4 and 5 was related to the presence of a free hydroxyl group at C-4′ since the 5-hydroxyl group was already blocked and the SC value of 5 with a free hydroxyl group at C-3 was relatively low.The antioxidant activity of three methylated quercetins empferol . The sam4 μg/mL) . The res1) and 3-hydroxy-5,7,4′-trimethoxyflavone (6), and four methylated quercetin methyl ethers: retusin  (4), 3,5-dihydroxy-7,3′,4′-trimethoxyflavone (5), 3,4′-dihydroxy-5,7,3′-trimethoxyflavone (7), and 3,5,7,3′,4′-pentamethoxyflavone (9), were isolated for the first time from the rhizomes of A. koenigii from Vietnam. Three methylated compounds 1, 4, and 5 were isolated from the fruit. The nonflavonoid compounds β-sitosterol (2), 5-hydroxy-1-(4′-hydroxyphenyl)eicosan-3-one (3), p-hydroquinone (8), and vanillic acid (10) were isolated for the first time from A. koenigii. Quantitative HPLC analysis revealed 1.81% of 1, 1.38% of 4, and 1.76% of 5 in dry rhizomes. In the antimicrobial activity test, except for the potent activity of 4 against A. niger with the MIC value of 100 μg/mL, compounds 1, 4, and 5 were not active at 100 μg/mL and 6 and 7 were not active at 50 μg/mL. Weak DPPH scavenging activity of three quercetin methyl ethers 4 (30.03%), 5 (14.20%), and 7 (42.61%) was observed at 100 μg/mL (4 and 5) or 50 μg/mL (7) concentration.Two kaempferol methyl ethers: 5-hydroxy-3,7,4′-trimethoxyflavone (


X-ray crystallography revealed the unintended formation of benzisothiazolinone 1-oxides from 1,3-benzothiazin-4-ones through oxidation instead of the anticipated benzothiazinone sulfones, which would be constitutional isomers. Mycobacterium tuberculosis, which have already reached clinical trials. A product obtained in low yield upon treatment of 8-nitro-2-(piperidin-1-yl)-6-(trifluoromethyl)-4H-benzothiazin-4-one with 3-chloroperbenzoic acid, in analogy to a literature report describing the formation of sulfoxide and sulfone derived from BTZ043 isothiazol-3(2H)-one 1-oxide, C14H12F3N3O5S, as revealed by X-ray crystallography. Single-crystal X-ray analysis of the oxidation product originally assigned as BTZ043 sulfone provides clear evidence that the structure of the purported BTZ043 sulfone is likewise the corresponding BIT 1-oxide, namely, 2-decane-8-carbonyl]-7-nitro-5-(trifluoromethyl)benzo[d]isothiazol-3(2H)-one 1-oxide, C17H16F3N3O7S. A possible mechanism for the ring contraction affording the BIT 1-oxides instead of the anticipated constitutionally isomeric BTZ sulfones and antimycobacterial activities thereof are discussed.1,3-Benzothiazin-4-ones (BTZs) are a promising new class of drugs with activity against  al. 2015. ACS Med Mycobacterium tuberculosis in vitro, 8-nitro-1,3-benzothiazin-4-ones (BTZs) have been the focus of many chemical, pharmacological and, recently, clinical studies  with 3-chloroperbenzoic acid in a similar way and crystallographic characterization of one of the oxidation products revealed the formation of a ring-contracted benzisothiazolone (BIT) 1-oxide instead of the anticipated BTZ sulfone -6-(trifluoromethyl)-4ne Fig. 1. Subsequ1 was synthesized as described elsewhere . The reported RF values are uncorrected. Flash chromatography was carried out with a 40 g puriFlash column . Preparative HPLC was performed on a Shimadzu LC-10AD system using 19 × 150 mm XTerra RP-18 columns . 1H and 13C NMR spectra were recorded at room temperature on an Agilent Technologies VNMRS 400 MHz NMR spectrometer . Chemical shifts are referenced to the residual signals of CDCl3 (δH = 7.26 ppm and δC = 77.0 ppm). High-resolution mass spectra (HRMS) were measured on a Bruker Daltonics APEXIII FT–ICR mass spectrometer.Starting materials were obtained from commercial sources and were used as received. Solvents were of analytical grade. Compound 2 and 3 were obtained when 1 was treated with 3-chloroperbenzoic acid, adapting the procedure described by Tiwari et al. , in dichloromethane (5 ml) at 0 °C. After stirring for 4 d at room temperature, additional 3-chloroperbenzoic acid (0.5 g) was added and the mixture was stirred for another day. The resulting mixture was washed twice with a saturated sodium bicarbonate solution (55 ml) and then once with deionized water (55 ml). After drying over sodium sulfate, the solvent was removed using a rotary evaporator. The crude product was subjected to flash chromatography [gradient of 50–100 (v/v) ethyl acetate/heptane] to give 2 and 3. Both compounds were purified by HPLC [gradient of 5–95 (v/v) acetonitrile/water in 10 min + 0.05% trifluoroacetic acid] to yield 6 mg of 2  and 35 mg of 3 .Compounds  al. 2015. A solut3 suitable for single-crystal X-ray analysis were obtained after a couple of days when a solution of ca 5 mg of the compound in ethanol (1.5 ml) in a 10 × 50 mm glass vial with a screw cap was left at room temperature and the solvent allowed to evaporate slowly.Crystals of 4 has been reported elsewhere : δ 8.77 , 8.57 , 3.58 , 1.78–1.70  ppm; HRMS(ESI): calculated for C14H12F3N3O4S [M + Na]+ 398.0398, found 398.0397; RF = 0.29 .1H NMR  δ 8.79 , 8.58 , 3.68–3.51 , 1.81–1.62  ppm; 13C NMR : δ 159.5, 148.3, 144.8, 143.0, 137.8 , 132.8, 129.4 , 126.6 , 121.6 , 47.5, 25.8, 24.0 ppm; HRMS(ESI): calculated for C14H12F3N3O5S [M + H]+ 392.0528, found 392.0526; RF = 0.22 .Uiso(H) = 1.2Ueq(C) (1.5 for methyl groups). The torsion angles of the methyl groups were initially determined using a circular Fourier search and subsequently refined while maintaining the tetrahedral structure.Crystal data, data collection and structure refinement details are summarized in Table 11 was treated with 3-chloroperbenzoic acid, adapting the procedure for BTZ043 reported by Tiwari et al. nitrobenzamide (see supporting information), following an established procedure for related BITs . Fig. 3R-configured at the S atom. It is worth emphasizing, however, that the S enantiomer is generated by glide symmetry in the polar crystal structure so the crystal is a racemate. The central carbamide moiety is tilted out of the BIT plane, as revealed by the torsion angles about the N2—C9 bond. The structure at atom N2 is slightly pyramidal, whereas that at N4 is virtually planar due to conjugation with the adjacent carbonyl group. The piperidine ring adopts a low-energy chair conformation with some deviations of the bond angles from ideal tetrahedal angles, which can be attributed to the planarity at N4.Compound 4 crystallizes in the Sohncke space group P21 with two diastereomers in the asymmetric unit (Z′ = 2). Fig. 43, compound 4 exhibits an additional spiro-(2S)-methyl-1,3-dioxolane group appended to the piperidine ring in the 4-position. The S configuration at C15, as in the BTZ043 starting material, is encountered in both crystallographically distinct molecules and the configurational assignment was confirmed by a Flack x parameter , the 1,3-dioxolane ring adopts an envelope conformation with atom C16 on the flap, whereas in molecule 2 , the ring is close to an envelope with the spiro atom C12 on the flap. As in 3, the BIT systems deviate slightly from planarity. In molecule 1, atom O2 is displaced from the mean plane of the benzene ring by 0.323 (6) Å, and in molecule 2, atoms N2 and O2 deviate by −0.118 (5) and 0.333 (5) Å, respectively, from this plane. The tilt angle between the mean plane of the benzene ring and the plane of the nitro group is 16.4 (3)° in molecule 1 and 10.7 (4)° in molecule 2. Similar to 3, in both molecules, the central carbamide moiety is tilted out of the plane of the BIT skeleton and the appended piperidine ring adopts a low-energy chair conformation with some minor deviations of the bond angles.Compound  1 Fig. 4a, the 1 2 Fig. 4b, the r4 in the solid state features short C—F⋯F—C contacts [F1_1⋯F3_2i = 2.737 (4) Å and F3_1⋯F1_2ii = 2.751 (4) Å)], which link unique molecules 1 and 2 along the [100] direction .The supramolecular structure of on Fig. 5. Accordiet al.  to the C=N bond of the BTZ system. Ring opening and rearrangement to a sulfenic acid group and an N-acylcarbamide moiety within the molecule would be followed by the loss of water to form 2, which was then oxidized by another equivalent of 3-chloroperbenzoic acid leading to 3, which we isolated and structurally characterized by X-ray crystallography. Although this mechanism is only postulated, it explains why both 2 and 3 were formed.Since the ring-contracted oxidation products only formed in very low yields, investigation of the reaction mechanism of BTZ oxidation and rearrangement upon treatment with 3-chloroperbenzoic acid was not undertaken. We propose the sequence shown in Fig. 6 al. 1988. We follet al. . Although the structures of 2 and 3 differ from those of ‘BTZ-SO’ and 4 by the absence of the spiro-(2S)-methyl-1,3-dioxolane group appended to the piperidine ring in the 4-position, their activities against M. tuberculosis and M. aurum are comparable  inhibitors, and work along this line is in progess. It should be noted, however, that BITs are known to have various molecular targets in microorganisms  global, 3, 4. DOI: 10.1107/S2053229620010931/ep30083sup2.hklStructure factors: contains datablock(s) 3. DOI: 10.1107/S2053229620010931/ep30084sup3.hklStructure factors: contains datablock(s) 4. DOI: Click here for additional data file.10.1107/S2053229620010931/ep30083sup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2053229620010931/ep30084sup5.cmlSupporting information file. DOI: 10.1107/S2053229620010931/ep3008sup6.pdfDescription of both the deliberate synthesis of 2 and the antimycobacterial assays. DOI: 2022514, 2022515CCDC references:


The crystal packing features intermolecular N—H⋯Cl and C—H⋯Cl hydrogen bonds and π–π stacking interactions.The title structure consists of isolated [SnCl H-imidazol-3-ium] hexachloridostannate(IV), (C6H9N2)2[SnCl6], has been prepared and characterized by single-crystal X-ray diffraction, Hirshfeld surface analysis and UV–visible spectroscopy. The structure consists of isolated [SnCl6]2− octahedral anions separated by layers of organic 1-(prop-2-en-1-yl)-1H-imidazol-3-ium cations. The 1-(prop-2-en-1-yl) fragment in the organic cation exhibits disorder over two sets of atomic sites having occupancies of 0.512 (9) and 0.488 (9). The crystal packing of the title compound is established by intermolecular N/C–H⋯Cl hydrogen bond and π– π stacking interactions. Hirshfeld surface analysis employing three-dimensional molecular surface contours and two-dimensional fingerprint plots has been used to analyse the intermolecular interactions present in the structure. The optical properties of the crystal were studied using UV–visible absorption spectroscopy, showing one intense band at 208 nm, which is attributed to π–π* transitions in the cations.A new 0D organic–inorganic hybrid material bis[1-(prop-2-en-1-yl)-1 The hexachlorostannate(IV) octahedron is nearly perfect with Sn—Cl bond lengths ranging from 2.4136 (6) to 2.4363 (6) Å and Cl—Sn—Cl bond angles between 88.44 (3) and 180°. These bond lengths and angles are in good agreement with those observed in similar compounds based on hexachlorostannate(IV) + cations are related to each other by 21/n symmetry elements. The overall negative charges in the structure are counter-balanced by the protonated 1-(prop-2-en-1-yl)-1H-imidazol-3-ium cations + organic cations.The asymmetric unit of  Å  to 1.180 (blue) a.u. The dnorm mapping indicates that strong hydrogen-bonding interactions, such as N—H⋯Cl hydrogen bonding between chlorine atoms and imidazolium groups and C—H⋯Cl hydrogen bonding between chlorine atoms and the hydrogen atoms of the 1-(prop-2-en-1-yl) groups, appear to be the primary interactions in the structure, seen as a bright-red area in the Hirshfeld surface 4[Bi4I16]·2H2O has been recently determined by Ferjani + organic cations.Optical absorption (OA) of the title compound was measured at ambient temperature in water. The experimental UV–visible absorption spectrum of the title compound is shown in Fig. 6H-imidazole] and 0.3 g (2 mmol) of tin(II) chloride in 10 ml of concentrated (37%) hydrochloric acid. The mixture was stirred with heating and then kept at room temperature. Three days later, colourless single crystals suitable for structural determination were obtained.The title compound was prepared by dissolving 0.34 g (1 mmol) of 1-allylimidazole [1-(prop-2-en-1-yl)-1A, C4B) and  with occupancy factors of 0.512 (9) and 0.488 (9). Geometrical restraints (SADI) on bond lengths were applied. H atoms were located in difference-Fourier maps but introduced in calculated positions and treated as riding on their parent atoms, with C—H = 0.93 and 0.97 Å, N—H = 0.86 Å with Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989020012177/vm2239sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020012177/vm2239Isup3.hklStructure factors: contains datablock(s) I. DOI: 2021940CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The online version of this article (10.1007/s13659-020-00265-x) contains supplementary material, which is available to authorized users. Cuscuta reflexa Roxb. (Convolvulaceae), a twining parasitic plant, is distributed in China, Afghanistan, India, Indonesia, Malaysia, Myanmar, Nepal, Pakistan, Sri Lanka, and Thailand , one methyl group , and one β-glucopyranosyl group [δH 5.05 ]. By comparing its NMR data with those of 4-(β-d-glucopyranosyloxy)-6-methyl-2H-pyran-2-one (5) , and one 2-methylbutyryl group  [2 was deduced as a 2-methylbutyryl derivative of 4-(β-glucopyranosyloxy)-6-methyl-2H-pyran-2-one by the COSY and HMBC correlations  was elucidated to be 4-[β-6-O-(2-methylbutyryl)-glucopyranosyloxy]-6-methyl-2H-pyran-2-one. We were unable to calculate the ECD spectrum of 2. The absolute configuration of C-2ʺ remains unknown.The molecular formula of cuscutaroside B (nd 91.6) , one β-g13C NMR ], one methoxy group , six methyl groups , and one trisubstituted double bond . Comparison of its NMR data with those of 7-oxo-β-sitosterol 3-O-β-d-glucopyranoside , and thus, H-7 is α-oriented. The ROESY correlations of H-1α/H-3, H-9/H-7, and H-7/H-14 indicated that H-3, H-9, and H-14 are also α-oriented. The configurations of C-20 and C-24 on the side chain could not be determined by the ROESY correlations. Because the NMR data of the side chain were highly consistent with those in the literature  to yield 13  and 14 .Part B was subjected to column chromatography  to yield one main subfraction. The 30% MeOH-eluted portion was isolated by Sephadex LH-20 (MeOH) and silica gel column chromatography  and further purified by semipreparative HPLC  to yield 15 .Fraction B2 was separated on an RP-C18 silica gel column eluted with MeOH-H2O (10% → 100%) to yield two main subfractions (B3-1 and B3-2). B3-1 (40% MeOH-eluted portion) was separated by silica gel column chromatography  to yield B3-1-1 and B3-1-2. B3-1-1 was separated by Sephadex LH-20 (MeOH) and silica gel column chromatography  followed by semipreparative HPLC  to yield 2  and 8 . B3-1-2 was separated by Sephadex LH-20 (MeOH) and silica gel column chromatography  to yield 1 (571.5 mg) and a mixture. The mixture was further purified by semipreparative HPLC  to yield 9 , 10 , and 11 . B3-2 (90% MeOH-eluted portion) was separated by Sephadex LH-20 column chromatography (MeOH) and semipreparative HPLC  to yield 4  and 3 .Fraction B3 was separated on an RP-C18 silica gel column eluted with MeOH–H2O (10% → 100%) to yield three main fractions. The 10% MeOH-eluted portion was purified by silica gel column chromatography  and Sephadex LH-20 column chromatography (MeOH) to yield 5 (1.1 g) recrystallized from MeOH. The 15% MeOH-eluted portion was isolated by silica gel column chromatography  and Sephadex LH-20 column chromatography (MeOH) and further purified by semipreparative HPLC  to yield 12 . The 20% MeOH-eluted portion was isolated by Sephadex LH-20 column chromatography (MeOH) and semipreparative HPLC  to yield 7 .Fraction B4 was separated on an RP-Cc = 0.19, MeOH); UV (MeOH) λmax (logε) 311 (4.42), 227 (4.11), 200 (4.49) nm; 1H NMR and 13C NMR data see Table m/z 433 [M-H]-, 867 [2 M-H]-; HRESIMS (positive) m/z 457.1106 [M + Na]+ .White powder; mentclass2pt{minimc = 0.11, MeOH); UV (MeOH) λmax (logε) 285 (3.54), 198 (4.22) nm; ECD  λmax (Δε) 281 (− 0.91), 199 (− 4.06) nm; 1H NMR and 13C NMR data see Table m/z 395 [M + Na]+, 767 [2M + Na]+; HRESIMS (positive) m/z 395.1313 [M + Na]+ .Colorless solid; mentclass2pt{minimc = 0.11, MeOH); 1H NMR and 13C NMR data see Table m/z 629 [M + Na]+; HRESIMS (positive) m/z 629.4388 [M + Na]+ .Colorless solid; entclass1pt{minima1  was dissolved in 15 mL of 6% aq. HCl and hydrolyzed under reflux (5 h) at 90 °C. Then, the acidic solution was evaporated in vacuo to dryness and separated by silica gel column chromatography eluted with CHCl3–MeOH (10:1) to yield d-glucopyranose , which was identified based on its 1H NMR spectrum and optical rotation value of [α]D20  + 40.0  +; HRESIMS (positive) m/z 625.1528 [M + Na]+ .Compound p-Nitrophenyl butyrate (p-NPB) was used as the substrate. First, 5 μL of the lipase solution (40 U/mL) in Tris–HCl buffer  was added to a 96-well microtiter plate. Each compound in 1 μL of DMSO and 184 μL of Tris–HCl buffer were added and mixed with the enzyme buffer to start the reaction. After incubation at 37 °C for 15 min, 10 μL of the substrate solution (10 mM p-NPB in dimethyl formamide) was added. The enzymatic reaction was carried out for 15 min at 37 °C. The hydrolysis of p-NPB to p-nitrophenol was monitored at 400 nm using a spectrophotometer [For lipase inhibition tests of each compound, porcine pancreatic lipase was used. otometer .The inhibitory effects of the compounds against rabbit platelet aggregation induced by thrombin (1 U/mL), PAF (0.4 μg/mL), AA (0.5 mM), or collagen (4 μg/mL) were evaluated according to published methods –30.Cuscuta reflexa (Convolvulaceae) collected from Myanmar. 7β-Methoxy-β-sitosterol 3-O-β-glucopyranoside (3) showed weak PPL inhibitory activity (IC50 = 67.2 ± 1.7 μg/mL). Cuscutaroside A (1), its acetylated derivative (1a), and scrophenoside B (8) showed weak inhibitory activity against rabbit platelet aggregation induced by collagen (4 μg/mL) with IC50 values of 291.4 ± 47.9 μg/mL, 63.8 ± 4.4 μg/mL, and 180.5 ± 6.7 μg/mL, respectively. Compound 1a also showed inhibitory activity against rabbit platelet aggregation induced by AA (0.5 mM) with an IC50 value of 72.6 ± 10.5 μg/mL. These results support previous findings in pharmacological studies related to the traditional uses of the plant.Three new and 12 known compounds were isolated from the whole plants of Supplementary file1 (PDF 1402 kb)Below is the link to the electronic supplementary material.


In the crystal, the molecules are linked by weak C—H⋯π hydrogen bonds and very weak π–π stacking interactions. Two-dimensional fingerprint plots show that the largest contributions to the crystal cohesion come from H⋯H and C⋯H/H⋯C interactions. 23H20N2OS, contains one slightly bent molecule. The naphthalene ring system and the thiazole ring are twisted with respect to each other, making a dihedral angle of 13.69 (10)°; the anisole ring is inclined to the plane of the naphthalene ring system, the dihedral angle being 14.22 (12)°. In the crystal structure, molecules are linked by C—H⋯π interactions, resulting in the formation of sheets parallel to (100). Within the sheets, very weak π–π stacking interactions lead to additional stabilization. Hirshfeld surface analysis and fingerprint plots reveal that the cohesion in the crystal structure is dominated by H⋯H (42.5%) and C⋯H/H⋯C (37.2%) contacts.The asymmetric unit of the title compound, C RN=CR2  and thus belongs to the family of imines , comprising a thiazole entity.Here we report on the synthesis, structure determination and Hirshfeld analysis of a Schiff base, CI) contains one molecule  Å, where Cg1 and Cg4 are the centroids of the S1/C12/C11/N1/C13 and the C15–C20 phenyl ring, respectively  by C4—H4⋯s Table 2, where Cly Fig. 3.et al., 2016viz. 4-(pyren-1-yl)-1,3-thiazol-2-amine , C19H12N2S -4-(4-chlorophenyl)-N--5--1,3-thiazol-2-amine, C19H12ClN5O2S  Å]. Distinctive bond lengths  in (I) are the same within standard deviations as the corresponding bond lengths in the structure of XAZJUE. In XAZJUE, the molecules are linked by weak C—H⋯O hydrogen bonds into a three-dimensional network.A search of the Cambridge Structural Database . Hirshfeld surface analysis was performed using a standard (high) surface resolution with the three-dimensional dnorm surfaces mapped over a fixed colour scale of −0.0638 (red) to 1.3242 (blue) a.u., and the results are illustrated in Fig. 4a. The red spots identified in Fig. 4a correspond to the near-type H⋯π contacts resulting from hydrogen bonds of the type C—H⋯π(ring)  contacts.Hirshfeld surface analysis  Table 2. The viea shows the two-dimensional fingerprint as the sum of all contacts contributing to the Hirshfeld surface indicated in normal mode. Fig. 5b illustrates the two-dimensional fingerprint of  points related to H⋯H contacts that represent a 42.5% contribution in the title structure. In Fig. 5c, two symmetrical wings on the left and right sides indicate C⋯H/H⋯C interactions with a contribution of 37.2%. Furthermore, there are S⋯H/H⋯S , N⋯H/H⋯N  and O⋯H/H⋯O  contacts contributing to the overall crystal packing of (I).Fig. 5I) was prepared by adding 4-N-propoxybenzaldehyde  dropwise under constant stirring to a solution of 2-amino-4-(2-naphthyl)thiazole  in 1-propanol (10 ml). The reaction was catalysed by NaOH (0.1 g), and the mixture stirred for 1 h in a water bath at approximately 278–283 K. The reaction was monitored by thin-layer chromatography (TLC) using ethyl acetate and n-hexane (3:7 v:v), and had an Rf of 0.675. The formed precipitate was filtered off, washed with 1-propanol, and dried. The resulting solid was further purified by washing with ethanol and diethyl ether. Single crystals of (I) for X-ray analysis were obtained by slow evaporation of an acetone solution .Compound (Uiso(H) = 1.5Ueq(C-methyl) or 1.2Ueq(C) for other C-bound H atoms.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989020006611/wm5556sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020006611/wm5556Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020006611/wm5556Isup3.cmlSupporting information file. DOI: 2004309CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


N-methyl-2-pyrrolidine are explored.The synthesis, structure, and theoretical studies of a calcium complex of a unique dianion derived from  catena-poly[[tetrakis(1-methylpyrrolidin-2-one-κO)calcium(II)]-μ-(E)-1,1′-dimethyl-2,2′-dioxo-1,1′,2,2′-tetrahydro--5,5′-bis(thiolato)-κ2O:O′], [Ca(C10H8N2O2S2)(C5H9NO)4]n, 1, crystallizes in the triclinic space group Pvia two different twofold operations, about the normals to (001) and (1E)-1,1′-dimethyl-2,2′-dioxo-1,1′,2,2′-tetrahydro--5,5′-bis(thiolate), [C10H8N2O2S2]2−, dianions (abbreviation: DMTBT). This dianion thus facilitates the formation of a 1-D polymer, which propagates in the [011] direction. These ribbons are linked by intermolecular C—H⋯S interactions. Each Ca atom is in an octahedral CaO6 six-coordinate environment with Ca—O bond lengths ranging from 2.308 (6) to 2.341 (6) Å, cis bond angles ranging from 88.2 (2) to 91.8 (2)° and the trans angles all 180° due to the Ca atoms being located on centers of inversion. Theoretical calculations were carried out using density functional theory (DFT) and the results showed that although the central DMTBT dianion is planar there is likely some resonance across the central bond between both azapentyl rings, but this is not sufficient to establish a ring current. The calculated UV–vis spectrum shows a peak at 625 nm, which accounts for the deep blue–purple color of solutions of the complex.The title compound,  An obstacle to such work is the lack of soluble alkaline earth thiolates. As part of a program for the investigation of precursors for the synthesis of a wide variety of metal sulfide materials, the reactions between alkaline earth amides and thiolate ligands were explored 2, was crystallized from a mixed NMP solution over a period of years, an unusual barium sulfur cluster was obtained, [Ba6(C4H9S)10S(C5H9NO)6], containing a central μ6-sulfido atom surrounded by six Ba atoms and NMP ligands  in N-methyl-2-pyrrolidone/calcium chloride solutions  to 2.341 (6) Å, cis bond angles ranging from 88.2 (2) to 91.8 (2)° and the trans angles all 180° due to the Ca atoms being located on centers of inversion. Thus each Ca atom has close to ideal octahedral geometry.The title compound, Cet al., 20141. However, there are no previous structures containing the dianion or any related species. This dianion has resulted from the condensation of two molecules of NMP along with the incorporation of two sulfur atoms in the form of C—S− bonds  within the The Ca atoms are located on centers of inversion. Each Ca is surrounded by 4 NMP ligands and coordinated through one of the two O atoms to two DMTBT dianions. This dianion thus facilitates the formation of 1-D ribbons, which propagate in the [011] direction. These ribbons are linked by C—H⋯S interactions Table 1, as showet al., 2016et al., 2014A search of the Cambridge Structural Database , with C—H distances ranging from 0.95 to 0.99 Å. The crystal was twinned by non-merohedry via two different twofold operations, about the normals to (001) and  I. DOI: 10.1107/S205698902001628X/pk2649Isup3.hklStructure factors: contains datablock(s) I. DOI: 2050442CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


S,4aR,8aR)-6-Oxo-2,4a,6,8a-tetrahydropyranopyran-2-carboxamide, a potentially effective antibacterial agent, was prepared by a chemoselective hydration of the corresponding nitrile using a heterogeneous catalytic method based on copper(II) supported on molecular sieves, in the presence of acetaldoxime. It belongs to a new class of pyranopyrans that possess antibacterial and phytotoxic activity.-6-oxo-2,4a,6,8a-tetrahydropyranopyran-2-carboxamide, C9H9NO4, 3, was prepared by a chemoselective hydration of the corresponding nitrile, 2, using a heterogeneous catalytic method based on copper(II) supported on molecular sieves, in the presence of acetaldoxime. Compound 3 belongs to a new class of pyranopyrans that possess antibacterial and phytotoxic activity. Crystallographic analysis of 3 shows a bent structure for the cis-fused bicyclic pyranopyran, similar to nitrile 2. Evidence of an intramolecular hydrogen bond involving the amide group and ring oxygen was not observed; however, two separate intermolecular hydrogen-bonding interactions were observed between the amide hydrogen atoms and adjacent carbonyl oxygen atoms along the b- and a-axis directions. The latter interaction may also be supported by an intermolecular C—H⋯O hydrogen bond. The lattice is filled out by close-packed layers of this hydrogen-bonded network along the c-axis direction, related from one to the next by a 21 screw axis.The pyranopyran amide (2 Compound 2 was also phytotoxic in an assay using the aquatic plant Lemna paucicostata (L.) Hegelm. (duckweed). As part of our ongoing efforts to synthesize additional analogs of 1 for testing as new antibacterials and herbicides, we have recently prepared amide 3, by a heterogeneous catalytic method that uses copper(II) supported on molecular sieves, in the presence of acetaldoxime to carry out chemoselective hydration of 2 ° versus 177° for a comparable trans-fused pyranopyran  carbonyl carbon in 3 at δ 174.1 ppm and the absence of the nitrile carbon that occurs at δ 114.9 in 2. The 1H spectrum of 3 shows slight changes in the chemical shifts of most protons, for example there is a downfield shift of H4A from δ 4.45 ppm in the nitrile to δ 4.61 ppm in the amide. The vinyl hydrogen H4 is also further downfield in the amide (δ 7.10 ppm vs 6.91 ppm). The torsion angle of 45.8° for H4A—C4A—C8A—H8A in 3 is consistent with the observed vicinal coupling constant of 4.5 Hz for H4A—H8A in the associated 1H NMR spectrum.The NMR spectra of the pyranopyran amide 3 are involved in intermolecular hydrogen bonding with adjacent carbonyl oxygen atoms: H1A with O2i and H1B with O3ii  x + 1, y, z.). A packing diagram of 3  shows the N—H⋯O hydrogen-bonding interactions forming molecular planes defined by the crystallographic a- and b-axes; packing of these hydrogen-bonded layers appears to be a function of solvent exclusion and van der Waals contact alone, lacking any hydrogen bonding.The amino hydrogen atoms of  3 Fig. 2a shows ab plane also presents an arrangement of C—H⋯O and C—H⋯π interactions that suggests two potential additional forces at play within the lattice of 3. Fig. 2b and Table 1ii and C6 and O3ii of adjacent copies of 3. These distances fall within parameters for C—H⋯O hydrogen bonding as has been described in well-characterized membrane proteins and peptidomimetics  are not consequences of an energetically dominant lattice arrangement and N—H⋯O hydrogen bonding, but rather have some measurable, albeit weak, energetic contribution to intermolecular association  is electrostatically matched with H4A . Proper exploration of this would require more advanced QM/MM methods, however, the crystal packing of 3 is at least suggestive of a favorable geometry and electrostatic environment for C—H⋯π interactions.Within the et al., 2016et al., 2019et al., 2019cis-fused and the two double bonds are in the same location as they are in 3. Among the total 40 structures that were found in the search, the pyranopyran core of several were trans-fused, for example, the bergenins and also truncated ladder ethers related to brevitoxin. Some compounds possessed aryl rings fused to the pyranopyran system while others had double bonds at alternate positions including the ring junction.A search of the Cambridge Structural Database -6-Oxo-2,4a,6,8a-tetrahydropyranopyran-2-carboxamide 3:3 was prepared by the procedure of Kiss & Hell -6-cyano-6,8a-dihydropyrano-pyran-2-(4aH)-one 2 , CuII-4 Å catalyst (0.022 g), acetaldoxime  and tert-butanol (2 mL) was stirred at 343 K for 4 h. The mixture was filtered through a pad of Celite and concentrated to a yellow–brown solid that was purified by cartridge chromatography on a Waters vacuum manifold system using 5% methanol/chloroform as eluant . Concentration of fractions left a white solid; yield, 0.0227 g (51.5%). Single crystals were obtained from a solution of 3 in 10% methanol/chloroform at 253 K. fR = 0.2 (10% methanol/ chloroform); mp 433-437 K; [α]D20 −268 ; IR (ATR) ν 3425, 3325, 3219, 1710, 1670, 1618 cm−1; 1H NMR  δ 7.10 , 6.40 , 6.16 , 6.10 , 4.86 , 4.80 , 4.61 ; 13C{1H} NMR (CDCl3) δ 174.1, 164.8, 143.7, 131.8, 124.5, 123.3, 74.0, 70.2, 64.4. HRMS (ESI–TOF) m/z calculated for C9H10NO4 196.0610, found 196.0607.Compound Hell 2011 with a cUiso(H) = 1.2Ueq(C).Crystal data, data collection, and structure refinement details are summarized in Table 210.1107/S2056989020001292/zl4038sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020001292/zl4038Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020001292/zl4038Isup3.cmlSupporting information file. DOI: 1980773CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Correction to: J Int Soc Sports Nutr 17, 45 (2020)https://doi.org/10.1186/s12970-020-00374-5Immediately after publication of this article , the autThe correct values in these sections are highlighted in bold below:‘Abstract’ section on page 20.18 kcal·min-1 greater than placebo at the post-beverage time point (p < 0.001) and 0.08 kcal·min-1 greater than placebo at the post-exercise time point (p = 0.011).Following consumption of the beverage, EE was ‘Energy expenditure’ in ‘Results’ section on page 60.07 kcal·min-1 from post-beverage to post-exercise (p = 0.025). Additionally, post-exercise EE was approximately 0.13 kcal·min-1 higher than the baseline (pre-beverage) EE (p < 0.001).However, EE increased by approximately 0.24 kcal·min-1 from pre-beverage to post-beverage (p < 0.001) and remained elevated by approximately 0.21 kcal·min-1 at post-exercise (p < 0.001).Following consumption of the energy drink, EE increased approximately 0.18 kcal·min-1 greater than placebo at the post-beverage time point (p < 0.001) and 0.08 kcal·min-1 greater than placebo at the post-exercise time point (p = 0.011).EE following consumption of the energy drink was ‘Discussion’ section on page 110.18 kcal·min-1 higher than placebo during the post-beverage time period, and approximately 0.08 kcal·min-1 higher than placebo during the post-exercise time period.Importantly, EE in the energy drink condition was found to be approximately The authors apologize for the inconvenience caused.