COMPARATIVE SEQUENCE ANALYSIS OF TLR2 TLR4 AND TLR9 GENES AMONG SELECTED VERTEBRATES-A META-ANALYSIS

Objective: Toll-like receptors are the pattern recognition receptors that recognize a diverse set of conserved pathogens. The receptors are also constantly under selection pressure because of the host antigen modifications. The present study focuses on how selection and mutation have modified the TLRs throughout the evolution in selected groups.Methods: We have selected the sequences of TLR2, 4 and 9 among Hominid group, Homo sapiens, Bubalus bubalis and Danio rerio in our analysis and analyzed different parameters like relative synonymous codon usage (RSCU), sequence divergence, amino acid composition and estimated evolutionary selection forces using Tajima's test.Results: The phylogenetic assessment proved that positive selection influences TLR2 and TLR4, but neutral selection/balancing selection occurred in TLR9 which concluded from the Tajima's test. Synonymous codon usage described the selection of leucine and arginine in all the sequences which describe the structural similarities of TLRs. Values of nucleotide pairs and disparity index proved the close relationship of Hominid and Human between TLR2 and TLR4 and TLR9 where the distant relationship was found with Danio. It can be hypothesized that some of the codons may be best selected for binding with the antigens and it was selected in the genome and some were eliminated due to selection pressure.Conclusion: The present study aimed to substantiate the closeness of TLR2 and TLR4 due to their functional similarity but distant with TLR9 because of the different antigens they recognized in the endosome

Role of toll like receptors in bacterial and viral diseases – A systemic approach

Background: Toll like receptors are key-receptors of the innate immune system, but their role against bacterial and viral infections are yet to be understood.Aim: The present study is aimed to investigate the diversity and frequency distribution of 10 TLR genes among typhoid fever and HIV+ patients. In this study, 44 samples were taken from typhoid patients and 55 samples from HIV+ patients.Patients and methods: Widal test positive samples (>1:80) in case of typhoid and the percentage of CD4+ count in case of HIV+ patient were considered for the PCR-SSP analysis.Results: We found that the frequencies of TLR1 and TLR6 were highest in typhoid patients, whereas the frequencies of TLR8 and TLR9 displayed higher among HIV+ patients. Chi-square values were significant for TLR8 and TLR10 in the case of typhoid patients, whereas in HIV patients significant values were considered for TLR2, TLR4, TLR8 and TLR9 respectively. The odds ratio calculated highest for TLR1 and TLR6 among typhoid patients. TLR4 and TLR9 calculated were highest odd for HIV+ patients. A door line association of TLRs with the disease was found when the relative risk was calculated for TLR2 (1.72), TLR3 (1.21) and TLR10 (1.98) in bacterial infection, whereas in case of viral infection relative risk was calculated for TLR4 (1.62), TLR8 (1.18) and in TLR9 (1.16).Conclusion: This study reports the frequency distribution and association of human TLR genes with the bacterial and viral infection in the North Bengal region of India for the first time. It also signified the gene- disease- environment association study in case of infectious diseases and also the risk factors of bacterial and viral infections in this region. It also depicts the role of TLRs in the recognition of the pathogens

A comparative cross‐platform meta‐analysis to identify potential biomarker genes common to endometriosis and recurrent pregnancy loss

Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.Endometriosis is characterized by unwanted growth of endometrial tissue in different locations of the female reproductive tract. It may lead to recurrent pregnancy loss, which is one of the worst curses for the reproductive age group of human populations around the world. Thus, there is an urgent need for unveiling any common source of origin of both these diseases and con-nections, if any. Herein, we aimed to identify common potential biomarker genes of these two diseases via in silico approach using meta‐analysis of microarray data. Datasets were selected for the study based on certain exclusion criteria. Those datasets were subjected to comparative meta‐anal-yses for the identification of differentially expressed genes (DEGs), that are common to both diag-noses. The DEGs were then subjected to protein‐protein networking and subsequent functional enrichment analyses for unveiling their role/function in connecting two diseases. From the analyses, 120 DEGs are reported to be significant out of which four genes have been found to be prominent. These include the CTNNB1, HNRNPAB, SNRPF and TWIST2 genes. The significantly enriched pathways based on the above‐mentioned genes are mainly centered on signaling and developmental events. These findings could significantly elucidate the underlying molecular events in endometri-osis‐based recurrent miscarriages.Peer reviewe

A Comparative Cross-Platform Analysis to Identify Potential Biomarker Genes for Evaluation of Teratozoospermia and Azoospermia

Publisher Copyright: © 2022 by the authors.Male infertility is a global public health concern. Teratozoospermia is a qualitative anomaly of spermatozoa morphology, contributing significantly to male infertility, whereas azoospermia is the complete absence of spermatozoa in the ejaculate. Thus, there is a serious need for unveiling the common origin and/or connection between both of these diseases, if any. This study aims to identify common potential biomarker genes of these two diseases via an in silico approach using a meta-analysis of microarray data. In this study, a differential expression analysis of genes was performed on four publicly available RNA microarray datasets, two each from teratozoospermia (GSE6872 and GSE6967) and azoospermia (GSE145467 and GSE25518). From the analysis, 118 DEGs were found to be common to teratozoospermia and azoospermia, and, interestingly, sperm autoantigenic protein 17 (SPA17) was found to possess the highest fold change value among all the DEGs (9.471), while coiled-coil domain-containing 90B (CCDC90B) and coiled-coil domain-containing 91 (CCDC91) genes were found to be common among three of analyses, i.e., Network Analyst, ExAtlas, and GEO2R. This observation indicates that SPA17, CCDC90B, and CCDC91 genes might have significant roles to play as potential biomarkers for teratozoospermia and azoospermia. Thus, our study opens a new window of research in this area and can provide an important theoretical basis for the diagnosis and treatment of both these diseases.Peer reviewe

Effects of CBLin depletion of intracellular ROS production generated by CCl₄ in WRL-68 cells.

<p>Production of ROS was measured by cleavage of acetate group of non-fluorescent H₂DCFDA (2',7'-dichlorodihydrofluorescein diacetate) which convert into DCF(2′,7′ -dichlorofluorescein) highly fluorescent. Cells were exposed to CCl₄ before treatment with CBL 50, 80, 100, 150 and 200 μg/ml for 24 h. The ROS production displays the intensity of fluorescence through the images of WRL-68 cells treated with different concentration of CBL <b>(D-H)</b>, CCl₄ <b>(C)</b>, H₂O₂ <b>(B)</b> and control <b>(A)</b>.</p

Amelioration of CCl₄ induced liver injury in swiss albino mice by antioxidant rich leaf extract of <i>Croton bonplandianus</i> Baill.

<div><p>The progress in industrialization has blessed mankind with a technologically superior lifestyle but poor management of industrial waste has in turn poisoned nature. One such chemical is carbon tetra chloride (CCl₄), which is a potent environmental toxin emitted from chemical industries and its presence in the atmosphere is increasing at an alarming rate. Presence of CCl₄ in human body is reported to cause liver damage through free radical mediated inflammatory processes. Kupffer cells present in the liver are potentially more sensitive to oxidative stress than hepatocytes. Kuffer cells produced tumor necrosis factor-α (TNF-α) in response to reactive oxygen species (ROS), that might further cause inflammation or apoptosis. In this study hepatoprotective capacity of antioxidant rich extract of <i>Croton bonplandianus</i> Baill. (CBL) was evaluated on CCl₄ induced acute hepatotoxicity in murine model. Hydro-methanolic extract of <i>C</i>. <i>bonplandianus</i> leaf was used for evaluation of free radical scavenging activity. Liver cells of experimental mice were damaged using CCl₄ and subsequently hepatoprotective potential of the plant extract was evaluated using series of <i>in-vivo</i> and <i>in-vitro</i> studies. In the hepatoprotective study, silymarin was used as a positive control. Antioxidant enzymes, pro-inflammatory markers, liver enzymatic and biochemical parameters were studied to evaluate hepatoprotective activity of <i>Croton bonplandianus</i> leaf extract. Free radical scavenging activity of CBL extract was also observed in WRL-68 cell line. The phytochemicals identified by GCMS analysis were scrutinized using <i>in-silico</i> molecular docking procedure. The results showed that CBL extract have potent free radical scavenging capacity. The biochemical parameters were over expressed due to CCl₄ administration, which were significantly normalized by CBL extract treatment. This finding was also supported by histopathological evidences showing less hepatocellularnecrosis, inflammation and fibrosis in CBL and silymarin treated group, compared to CCl₄ group. ROS generated due to H₂O₂ in WRL-68 cell line were normalize in the highest group (200 μg/ml) when compared with control and negative control (CCl₄) group. After molecular docking analysis, it was observed that the compound α-amyrin present in the leaf extract of <i>C</i>. <i>bonplandianus</i> has better potentiality to protect hepatocellular damages than the standard drug Silymarin. The present study provided supportive evidence that CBL extract possesses potent hepatoprotective capacity by ameliorating haloalkane induced liver injury in the murine model. The antioxidant and anti-inflammatory activities also affirm the same. The synergistic effects of the phytochemicals present in CBL are to be credited for all the hepatoprotective activity claimed above.</p></div

Gas Chromatography Mass Spectrometry (GC-MS) analysis of <i>Croton bonplandianus</i>.

<p>Gas Chromatography Mass Spectrometry (GC-MS) analysis of <i>Croton bonplandianus</i>.</p

Molecular docking (secondary structure view) between Hepatitis BX protein and α-amyrin.

<p>Molecular docking (secondary structure view) between Hepatitis BX protein and α-amyrin.</p

Antioxidant activity of <i>Croton bonplandianus</i>.

<p><b>(A)</b> % inhibition of Singlet O₂ (¹O₂) Vs standard lipoic acid; <b>(B)</b> depicts unneutralized Singlet O₂ (¹O₂); <b>(C)</b> % inhibition of Nitric oxide (NO) Vs standard curcumin; <b>(D)</b> depicts unneutralized nitric oxide (NO). Data expressed as mean ± S.D (n = 6). α p<0.05; β p<0.01; γ p<0.001; NS-Non significant when compared with standard.</p

Antioxidant activity of <i>Croton bonplandianus</i>.

<p><b>(A)</b> % inhibition of Peroxynitrite (OONOˉ) Vs standard gallic acid; <b>(B)</b> depicts unneutralized Peroxynitrite (OONOˉ); <b>(C)</b> Total reducing power activity.Data expressed as mean ± S.D (n = 6). α p<0.05; β p<0.01; γ p<0.001; NS-Non significant when compared with standard.</p