Chemical leukoderma: An insight of pathophysiology and contributing factors.

Chemical leukoderma, or chemical-based vitiligo, is a dermal disease triggered by exposure to chemicals and characterized by the emergence of depigmentation or hypopigmentation of the skin. The etiology of this condition is associated with exposure to various chemical substances present in both occupational and non-occupational settings. The precise mechanism that underlies chemical leukoderma remains elusive and is believed to result from the demise of melanocytes, which are responsible for producing skin pigments. This condition has gained particular prominence in developing countries like India. An interesting connection between chemical leukoderma and vitiligo has been identified; studies suggest that exposure to many household chemicals, which are derivatives of phenols and catechol, may serve as a primary etiological factor for the condition. Similar to autoimmune diseases, its pathogenesis involves contributions from both genetic and environmental factors. Furthermore, over the last few decades, various studies have demonstrated that exposure to chemicals plays a crucial role in initiating and progressing chemical leukoderma, including cases stemming from occupational exposure.

Population genetic structure of the malaria vector Anopheles fluviatilis species T (Diptera: Culicidae) in India.

Anopheles fluviatilis James (Diptera: Culicidae) represents a complex that comprises four sibling species (S, T, U, and V). Among these, species T is widely distributed in India. Chromosomal inversion polymorphism exists among different geographic populations of An. fluviatilis species T; however, population genetic structure is not understood. This study inferred a genetic structure among six geographically diverse populations of species T using a panel of microsatellite markers. Analyses indicated a significant but low genetic differentiation among the majority of the studied populations. A significant correlation was observed between genetic and geographic distances, exhibiting stepwise migration patterns among populations.

Epitope based peptide vaccine against SARS-COV2: an immune-informatics approach.

World is witnessing exponential growth of SARS-CoV2 and fatal outcomes of COVID 19 has proved its pandemic potential already by claiming more than 3 lakhs deaths globally. If not controlled, this ongoing pandemic can cause irreparable socio-economic and psychological impact worldwide. Therefore a safe and effective vaccine against COVID 19 is exigent. Recent advances in immunoinformatics approaches could potentially decline the attrition rate and accelerate the process of vaccine development in these unprecedented times. In the present study, a multivalent subunit vaccine targeting S2 subunit of the SARS-CoV2 S glycoprotein has been designed using open source, immunoinformatics tools. Designed construct comprises of epitopes capable of inducing T cell, B cell (Linear and discontinuous) and Interferon γ. physiologically, vaccine construct is predicted to be thermostable, antigenic, immunogenic, non allergen and non toxic in nature. According to population coverage analysis, designed multiepitope vaccine covers 99.26% population globally. 3D structure of vaccine construct was designed, validated and refined to obtain high quality structure. Refined structure was docked against Toll like receptors to confirm the interactions between them. Vaccine peptide sequence was reverse transcribed, codon optimized and cloned in pET vector. Our in-silico study suggests that proposed vaccine against fusion domain of virus has the potential to elicit an innate as well as humoral immune response in human and restrict the entry of virus inside the cell. Results of the study offer a framework for in-vivo analysis that may hasten the process of development of therapeutic tools against COVID 19.Communicated by Ramaswamy H. Sarma.

Cross-sectional study of the prevalence of polycystic ovary syndrome in rural and urban populations.

OBJECTIVE: To assess the prevalence and risk factor profile of polycystic ovary syndrome (PCOS) in Haryana, India. METHODS: A large-scale cross-sectional study was conducted among women of reproductive age in Haryana between December 2015 and May 2017. A random multi-stage stratified sampling method was adopted. PCOS screening was based on questionnaires. Blood samples for hormonal analysis were collected from those with probable and definitive PCOS cases. Women with menstrual irregularities (MI), hyperandrogenism (HA), and polycystic ovaries (PCO) (Rotterdam criteria) were included. Females with thyroid disease, hyperprolactinemia, and adrenal hyperplasia were excluded. RESULTS: Among total 2400 women screened, 94 (4.21%) had PCOS. The PCOS phenotypes were 30% clinical HA (hirsutism, H), 64% biochemical HA, 35% PCO, 16% H+MI, 10% MI+PCO, 52% MI+HA, 14% PCO+H, and 19% PCO+H+HA. Overall, 67 (71%) of the women with PCOS resided in urban regions and 27 (29%) in rural regions. CONCLUSION: Among the women with PCOS, a considerably higher proportion resided in urban regions of Haryana. The difference may be attributed to lifestyle and dietary factors. Ignoring PCOS may put women at risk of serious long-term health consequences that are difficult to manage. Lifestyle changes and continuous surveys should be promoted for better management.

Association of rs6259 polymorphism with SHBG levels and Poly Cystic Ovary Syndrome in Indian population: a case control study.

Polycystic ovary syndrome (PCOS) is the most common endocrinopathy of reproductive-aged women. PCOS reflects a number of possible etiologies but its pathophysiology is still unclear. The principal abnormality of the syndrome is hyperandrogenism (70-80%). The access of androgens to target tissues is regulated by sex hormone-binding globulin (SHBG), a transport protein secreted by liver i.e. specific for androgens. Present study was done to find the association of rs6259 polymorphism with SHBG levels and Poly Cystic Ovary Syndrome in Indian population. Present study was a case control study. 400 subjects were enrolled for the study and serum SHBG levels and D327N polymorphism were measured. The D327N polymorphism (wild-type and variant allele) was detected using PCR-RFLP method (restriction enzyme Bbs-I). PCOS group was found to have significantly lower SHBG levels than healthy controls. There was no significant difference in genotype distribution between PCOS and controls (χ2 = 1.0335, p = 0.59). Significant difference in SHBG levels of PCOS and control group highlights the potential of SHBG as a biomarker for PCOS. However, no significant difference in genotype distribution between PCOS and controls provided an insight that rs6259 polymorphism is not associated with the risk of PCOS and SHBG levels.

Evaluating the association of TNF α promoter haplotype with its serum levels and the risk of PCOS: A case control study.

Poly cystic ovary syndrome is the major cause of anovulatory infertility. TNF α, pro-inflammatory cytokine is associated with obesity, insulin resistance and hyperandrogenism, therefore in present study we tried to find the association between TNF α promoter polymorphisms, TNF α levels and the risk of PCOS. Present case control study was carried on 400 women of age 16-40 years. TNF α levels were measured by ELISA whereas promoter polymorphisms were evaluated by PCR-RFLP. Haplotype and Linkage disequilibrium analysis was also done. TNF α level was significantly higher in PCOS group (13.24 ±â€¯9.78) than control (5.5 ±â€¯3.8). Haplotype analysis revealed that GGTCT, AGTCT, AGCCT and AACCT are the susceptible haplotypes associated with TNF α level. rs361525 and rs1799964 were found to be associated with the risk of PCOS (p = 0.0006, 0.015). GGCCT, AATAT, GATAT (most susceptible), AGCCT, GGTCT and GATCT are the susceptible haplotypes for PCOS. Significant difference between TNF α levels in PCOS and Control group suggest it's one of the promising candidates for the marker of inflammation (sensitivity and specificity 91.23 and 94.56% respectively, with area under the curve 0.907 at 95% CI 0.8723-0.9512). Presence of GGCCT haplotype suggests the susceptibility towards PCOS which needs to be further verified. In addition to this, present study not only provides a pavement for the diagnosis, but also monitoring and management of PCOS too.

Comprehensive analysis of damage associated SNPs of MMP9 gene: A computational approach.

Extracellular matrix is a dynamic meshwork of macromolecules that plays an important role in biological processes such as tissue remodeling and various developmental processes. Collagen is the chief component of ECM. Upon hydrolysis, it forms an irreversible left-handed helical structure which is further hydrolyzed by a specialized group of MMP family i.e. Gelatinases (MMP2 and MMP9). Present study was carried to figure out deleterious SNPs associated with MMP9 gene. Our results showed that two nsSNP (rs8125581 and rs41529445) that are present in catalytic domain are highly conserved and affect the protein structure and function.7 SNPs located in UTRs were found to alter miRNA seed region 13 SNPs of splice site were predicted to affect splice signals thereby affecting the post translational expression of MMP9. Most of the SNPs are still uncharacterized thereby present study provides a direction that can help to validate the relation between the altered expressions and functions of MMP9 protein in terms of disease susceptibility.

Comprehensive in-silico prediction of damage associated SNPs in Human Prolidase gene.

Prolidase is cytosolic manganese dependent exopeptidase responsible for the catabolism of imido di and tripeptides. Prolidase levels have been associated with a number of diseases such as bipolar disorder, erectile dysfunction and varied cancers. Single nucleotide polymorphism present in coding region of proteins (nsSNPs) has the potential to alter the primary structure as well as function of the protein. Hence, it becomes necessary to differentiate the potential harmful nsSNPs from the neutral ones. 19 nsSNPs were predicted as damaging by in-silico analysis of 298 nsSNPs retrieved from dbSNP database. Consurf analysis showed 18 out of 19 substitutions were present in the conserved regions. 4 substitutions (D276N, D287N, E412K, and G448R) that observed to have damaging effect are present in catalytic pocket. Four SNPs listed in splice site region were found to affect splicing of mRNA by altering acceptor site. On 3'UTR scan of 77 SNPs listed in SNP database, 9 SNPs were lead to alter miRNA target sites. These results provide a filtered data to explore the effect of uncharacterized nsSNP and SNP related to UTRs and splice site of prolidase to find their association with the disease susceptibility and to design the target dependent drugs for therapeutics.

Plasma prolidase levels as a biomarker for polycystic ovary syndrome.

AIM: Assessment of plasma prolidase levels in polycystic ovary syndrome (PCOS). PATIENTS & METHODS: PCOS patients were screened according to Rotterdam Criterion and prolidase levels were measured. RESULTS: A total of 170 patients and 160 controls were recruited for the study and it was found that prolidase levels were significantly higher in PCOS group (991.10 ± 39.52) than control (621.89 ± 23.94). Furthermore it has been found that prolidase levels increase with the number of cysts in ovaries. CONCLUSION: Significant difference between prolidase levels in PCOS and control shows that it may be used as a diagnostic marker for disease. In addition to this, there is a positive correlation found between prolidase levels and number of cysts, hence may be used as a prognostic marker to monitor disease status.

Disagreement in genotyping results of drug resistance alleles of the Plasmodium falciparum dihydrofolate reductase (Pfdhfr) gene by allele-specific PCR (ASPCR) assays and Sanger sequencing.

The rapid spread of antimalarial drug resistance in Plasmodium falciparum over the past few decades has necessitated intensive monitoring of such resistance for an effective malaria control strategy. P. falciparum dihydropteroate synthase (Pfdhps) and P. falciparum dihydrofolate reductase (Pfdhfr) genes act as molecular markers for resistance against the antimalarial drugs sulphadoxine and pyrimethamine, respectively. Resistance to pyrimethamine which is used as a partner drug in artemisinin combination therapy (ACT) is associated with several mutations in the Pfdhfr gene, namely A16V, N51I, C59R, S108N/T and I164L. Therefore, routine monitoring of Pfdhfr-drug-resistant alleles in a population may help in effective drug resistance management. Allele-specific PCR (ASPCR) is one of the commonly used methods for molecular genotyping of these alleles. In this study, we genotyped 55 samples of P. falciparum for allele discrimination at four codons of Pfdhfr (N51, C59, S108 and I164) by ASPCR using published methods and by Sanger's DNA sequencing method. We found that the ASPCR identified a significantly higher number of mutant alleles as compared to the DNA sequencing method. Such discrepancies arise due to the non-specificity of some of the allele-specific primer sets and due to the lack of sensitivity of Sanger's DNA sequencing method to detect minor alleles present in multiple clone infections. This study reveals the need of a highly specific and sensitive method for genotyping and detecting minor drug-resistant alleles present in multiple clonal infections.

Isolation and Characterization of Polymorphic Microsatellite Markers from the Malaria Vector Anopheles fluviatilis Species T (Diptera: Culicidae).

Anopheles fluviatilis James is an important malaria vector in India, Pakistan, Nepal, and Iran. It has now been recognized as a complex of at least four sibling species-S, T, U, and V, among which species T is the most widely distributed species throughout India. The taxonomic status of these species is confusing owing to controversies prevailing in the literature. In addition, chromosomal inversion genotypes, which were considered species-diagnostic for An. fluviatilis species T, are unreliable due to the existence of polymorphism in some populations. To study the genetic diversity at population level, we isolated and characterized 20 microsatellite markers from microsatellite-enriched genomic DNA library of An. fluviatilis T, of which 18 were polymorphic while two were monomorphic. The number of alleles per locus among polymorphic markers ranged from 4 to 19, and values for observed and expected heterozygosities varied from 0.352 to 0.857 and from 0.575 to 0.933, respectively. Thirteen markers had cross-cryptic species transferability to species S and U of the Fluviatilis Complex. This study provides a promising genetic tool for the population genetic analyses of An. fluviatilis.

Polymorphism in drug resistance genes dihydrofolate reductase and dihydropteroate synthase in Plasmodium falciparum in some states of India.

BACKGROUND: Sulfadoxine-pyrimethamine (SP) combination drug is currently being used in India for the treatment of Plasmodium falciparum as partner drug in artemisinin-based combination therapy (ACT). Resistance to sulfadoxine and pyrimethamine in P. falciparum is linked with mutations in dihydropteroate synthase (pfdhps) and dihydrofolate reductase (pfdhfr) genes respectively. This study was undertaken to estimate the prevalence of such mutations in pfdhfr and pfdhps genes in four states of India. METHODS: Plasmodium falciparum isolates were collected from two states of India with high malaria incidence i.e., Jharkhand and Odisha and two states with low malaria incidence i.e., Andhra Pradesh and Uttar Pradesh between years 2006 to 2012. Part of sulfadoxine-pyrimethamine (SP) drug resistance genes, pfdhfr and pfdhps were PCR-amplified, sequenced and analyzed. RESULTS: A total of 217 confirmed P. falciparum isolates were sequenced for both Pfdhfr and pfdhps gene. Two pfdhfr mutations 59R and 108N were most common mutations prevalent in all localities in 77 % of isolates. Additionally, I164L was found in Odisha and Jharkhand only (4/70 and 8/84, respectively). Another mutation 51I was found in Odisha only (3/70). The pfdhps mutations 436A, 437G, 540E and 581G were found in Jharkhand and Odisha only in 13, 26, 14 and 13 % isolates respectively, and was absent in Uttar Pradesh and Andhra Pradesh. Combined together for pfdhps and pfdhfr locus, triple, quadruple, quintuple and sextuple mutations were present in Jharkhand and Odisha while absent in Uttar Pradesh and Andhra Pradesh. CONCLUSION: While only double mutants of pfdhfr was present in low transmission area (Uttar Pradesh and Andhra Pradesh) with total absence of pfdhps mutants, up to sextuple mutations were present in high transmission areas (Odisha and Jharkhand) for both the genes combined. Presence of multiple mutations in pfdhfr and pfdhps genes linked to SP resistance in high transmission area may lead to fixation of multiple mutations in presence of high drug pressure and high recombination rate.