Immunotoxicity of cynobacterial toxin Microcystin-LR is mitigated by Quercitin and himalaya tonic Liv52.

Microcystin-LR (MC-LR) has received worldwide concern for its hepatotoxicity with maximum acceptable daily intake of 0.0015 mg/L (1.5 µg/L) [Federal-Provinicial-Territorial-Committee-on-drinking-water-2002]. Comprehensive immunotoxicity data is still deficient with MC-LR. To curb the menace of MC-LR, Quercitin (QE), himalaya made hepatotonic Liv52 were studied. To investigate the immunotoxic properties of MC-LR, QE and Liv52, primary splenocyte cells prepared, cultured, and immunoproliferation assay with mitogens lipopolysaccharide (LPS) or concanavalin A, (Con A) was done for, immunophenotyping, cell cycle and apoptotic studies. In current study, we have divided the splenocytes into 4 groups, i.e., Group I: Normal saline, Group II: MC-LR (0.1 µM), Group III: MC-LR (0.1 µM) + QE (20 µM), and Group IV: MC-LR (0.1 µM) + Liv52 (25 µg/ml) and treated with maximum < CC50 concentration. MC-LR enhanced proliferation of Con A and LPS stirred splenocytes at 24 h, whereas QE and Liv52 both act as antimitogenic. With combined mixture of MC-LR + QE, a significant increase in proliferation compared to mitogen or MC-LR was observed. MC-LR down-regulated expression of CD19+, CD3e+, CD4+, CD8+, (1.05%), (18.9%), (8.9%), and (7.8%) respectively in comparison to Group I. Down-regulation of 10% and 28% is observed in CD19+ and CD4+ populations with MC-LR and QE. The Liv52 addition concealed MC-LR adverse properties in most effective way. MC-LR induced G1-phase significant declined cell cycle arrest at S phase (9.26%) and G2/M phase (26.31%) was observed. QE and Liv52 mask the activity of MC-LR. Further apoptotic study revealed that MC-LR treatment decreases late apoptotic cells compared to control with no significant change in live and early apoptotic cells. Although QE increased live cells and Liv52 significantly increased late apoptotic cells, these results suggest that a

Diagnosis of Genital Tuberculosis in Infertile Women by Using the Composite Reference Standard.

Female genital tuberculosis (FGTB) can be asymptomatic or even masquerade as other gynecological conditions. Conventional methods of FGTB diagnosis include various imaging, bacteriological, molecular, and pathological techniques that are only positive in a small percentage of patients, leaving many cases with undiagnosed condition. In the absence of a perfect diagnostic method, composite reference standards (CRSs) have been advocated in this diagnostic study. This study assesses the agreement between traditional diagnostic modalities using CRS and prevalent TB groups among different fallopian tube infertility manifestations. A total of 86 women with primary and secondary infertility were included in the study and subjected to bacteriological, pathological, and radiological examination for the diagnosis of FGTB. Results were evaluated statistically for concordance of the diagnostic tests to the CRS by sensitivity and specificity, while PPV and NPV were calculated for the performance of diagnostic tests of FGTB. We observed that 11.2% of women were found to be true positives by means of CRS. The positive findings by CRS were as follows: ultrasonography (13.9%), laparoscopy (14%), hysteroscopy (12%), GeneXpert (4.8%), culture (4.8%), polymerase chain reaction (4.8%), and histopathology (6.4%). GeneXpert and culture were found to have a perfect agreement with CRS. Hysterosalpingography, laparoscopy, and hysteroscopy have a fair agreement with CRS. Out of 43 women with tubal factor infertility, 6 women were found in the definitive TB group with mixed conditions of tubal manifestations. This study evaluates and demonstrates the reliability of the collective assessment of various diagnostic methods with CRS findings that help in identifying different TB groups of genital tuberculosis patients from all infertile patients by applying the criteria of CRS.

Molecular modelling and docking of Mus musculus HMGB1 inflammatory protein with CGA.

Recently, High Mobility Group Box1 (HMGB1) protein has been reported as an inflammatory cytokine present in all nucleated cells with crucial role in the genesis and promotion of cancer. No HMGB1 protein mice model and its active site details are available to validate mice in vivo experiments. Here, for the first time we have reported in silico mice HMGB1 model using human HMGB1 template. Prepared HMGB1 secondary structure showed 6-α helices, 5-ß turns, 2-γ turns with 67% α-helices, 32% coil and 9% turn without ß-sheet, and classified as α-class protein. Ramachandran plot analysis showed 98.2% and 92.3% residues lies in favoured region, verified by RAMPAGE and PDBsum server respectively. Cancer atlas of HMGB1 protein showed up-regulated expression of HMGB1 gene in different cancer, proved by CAB (CAB005873) and HPA-antibody (HPA003506) in silico. HMGB1 protein showed interaction with different biologically important inflammatory protein as depicted in STRING result.Prominent active site has residues Tyr78Ile79Pro80-81Lys82Gly83vGlu84Thr85Lys86-88Phe89Lys90Asp91Pro92Asn93Tyr162Lys165 with 310 Å3 site volume.Interacting residues of CGA-HMGB1 docked complex were ILE79PRO80-81LYS82GLY83GLU84LYS86-88PHE89Arg163Ala164LYS165Gly166 with docking score 3872 and surface area 412.6. CGA-conformer C3950 showed best docking than CGA and conformer-ZINC03947476, iso-chlorogenic acid and cischlorogenic acid. HMGB1 mice model could be a good therapeutic target for anti-cancerous drugs.

HMGB1 protein as a novel target for cancer.

Highly conserved nuclear protein High Mobility Group Box1 (HMGB1) present in mammals has functionality as an immuno-modulator in the form of cytokine molecule, as a nuclear factor to regulate these molecules and DNA structural determination. It has proximal homologous DNA binding domains Box-A, Box-B and distal C-terminal domain. Reduced form exists in basic condition has chemotaxis activity, while form with disulphide bond reduced at 106th cysteine showed cytokine activity. The oxidized form is devoid of both activities. HMGB1 binds and bends dsDNA and also activates genes for secretion of inflammatory cytokines such as IL-1ß, TNF-α, IL-6 and IL-18. It can interact with transcription factors Rel/NF-κB and p53 responsible for up-regulating oncogenes. Oxidative stressed injured tissues actively secrete HMGB1 outside cells to necrotize other nearby tissues passively in cytosol. Acetylation of HMGB1 weakens its binding with DNA, and promotes its migration to different tissues leading to secretion of inflammatory-cytokines. HMGB1 expression has been found very important in the genesis and promotion of different cancer by promoting metastasis. In current article, we emphasized on condition based structural variability of HMGB1, mechanism of release, physiological functions and its functionality as a biomarker for cancer to be targeted to curb cancer genesis and progression.