Whole exome and targeted sequencing reveal novel mutations associated with inherited PCOS condition in an Indian cohort.

A cohort of polycystic ovary syndrome (PCOS) women presents themselves with persistent abnormal reproductive hormone levels and has a familial representation of characteristics. In our study, we have aimed to identify genetic variants which are inherited across such PCOS families and also validate them among Indian population. Independent discovery was done by whole exome sequencing in a three-generation family (Family P01). Validation was done by targeted sequencing at 30,000x using HaloPlex panel in 9 families (P01-P09). The variants were filtered and reported according to American College of Medical Genetics and Genomics (ACMG) guidelines. Mutation burden analysis and in-silico functional analyses were performed. After careful annotation analyses, we report 24 likely pathogenic variants from 21 genes, out of which 8 are novel structural variants, 14 missense variants and 2 intronic variants. Out of these, 3 variants from the genes FSHR, SCARB1, and INSR are involved in the ovarian steroidogenesis pathway and 5 variants from genes DFFB, ACTG1, GPX4, CYC1 and ALDOA directly or indirectly trigger the apoptotic pathways. Three ovarian steroidogenesis variants, FSHR, SCARB1 and INSR were screened among Indian women using a case-control approach to validate these variant's pathogenicity in Indian PCOS women. Variants of SCARB1 and INSR were found to be pathogenic to Indian PCOS women, while FSHR variants did not show significant association to PCOS cases.

Caesalpinia bonduc (L.) Roxb. as a promising source of pharmacological compounds to treat Poly Cystic Ovary Syndrome (PCOS): A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Caesalpinia bonduc (L.) Roxb. [Family: Fabaceae (Caesalpiniaceae)], known as bonduc nut or fever nut, has been used in India for many years to treat fever, inflammation, diabetes, cardiovascular disorder, cancer and also for birth control. In recent years, the seeds of this plant are consumed to regulate the menstrual disorders in PCOS. Poly cystic ovary syndrome (PCOS) is one of the most common endocrinological disorders faced by women of reproductive-age in several countries. AIM OF THE REVIEW: This review is aimed at providing the updates on the potential of Caesalpinia bonducella in treating the major symptoms of PCOS and also highlights the future research needs. MATERIALS AND METHODS: All the data pertaining to the plant was collected from multiple databases like Google, Science Direct, Springer, PubMed, Reaxys, Sci Finder, Wiley Online library and Taylor and Francis without any language restrictions. Moreover, information related to PCOS treatment was obtained from native medicine practitioners and from Ayurvedic pharmacopeia of India. RESULTS: More than 50 compounds have been identified in the seed kernels of C. bonducella so far. Seed kernel extracts rich in active compounds like bonducellin, caesalpinin, cassane furanoditerpenes, terpernoids, flavonoids and sterols are studied for their anti-diabetic, anti-hyperlipidemic, anti-cancer, anti-pyretic, and anti-oxidant properties. Recent literature has also attributed anti-androgenic and anti-estrogenic potential to this plant which might be helpful in regulating hyperandrogenism, the major predisposing factor contributing to various other clinical symptoms of PCOS. CONCLUSION: Phytochemicals in C. bonducella are effective against several ailments attributed to PCOS. However, there are no experiments done in PCOS animal models to claim the direct application of C. bonducella in PCOS treatment. Hence, further dose dependent toxicity studies are warranted in animal PCOS models and in PCOS patients to support the traditional evidence of the plants' potential to treat PCOS.

Rhizomicrobiomics of Caesalpinia bonducella, a wonder plant for PCOS treatment.

Plant and rhizobacterial interactions contribute partly to a plant's medicinal properties and are well studied through metagenomics. In this study, 16S rDNA, 18S rDNA, and ITS meta-sequencing were performed using the genomic DNA obtained from the rhizosphere of Caesalpinia bonducella-a medicinal shrub widely used to treat polycystic ovary syndrome (PCOS). Of the 665 Operational Taxonomic Units (OTUs) obtained from 16S rDNA sequencing, 23.9% comprised of microbes that increase the therapeutic value of plants (Bacillus, Paenibacillus), 6.4% belonged to stress and drought tolerant microbes (Pseudomonas, Rhizobium, Serratia), 8% belonged to plant-growth promoting rhizobacteria-predominantly Proteobacteria, and Firmicutes and the remaining were the microbes performing various other functions. Alpha diversity indexing by GAIA-metagenomics tool revealed the presence of a highly diverse group of microbes in the rhizosphere of C. bonducella; Chao.1 index (665), Shannon Weiner index (3.53), Simpson index (0.83) and Fisher index (106.13). The highly diverse microbes lingering around the roots of C. bonducella could possibly be due to a strong symbiotic association with the plant; root exudates nourish the microbes and the microbes in turn enrich the medicinal value of the plant.

Association of GWAS identified INSR variants (rs2059807 & rs1799817) with polycystic ovarian syndrome in Indian women.

Polycystic ovary syndrome (PCOS), a gynaecological endocrine disorder affects 9% of Indian women and is linked to type II diabetes. The association of INSR (INSulin Receptor gene) variants (rs2059807 and rs1799817) with PCOS was established through genome-wide association studies, yet requires validation for the Indian population. This case-control study included 253 PCOS women and 308 age-matched control. The minor allele frequency of rs2059807 had an odds ratio of 13.5 and that of rs1799817 was 11.8. The cohort with rs2059807 MAF presented elevated levels of luteinising hormone [PCOS vs Control: 6.32 ± 2.26 mIU/mL vs 4.97 ± 3.27 mIU/mL], estradiol [116.01 ± 60.63 pg/mL vs 65.04 ± 44.98 pg/mL], and decreased HDL - C [50.4 ± 11.59 mg/dL vs 64 ± 15.49 mg/dL] showing disturbances in the hormonal patterns. The rs1799817 polymorphism cohort had elevated levels of serum insulin [17.99 ± 11.6 mIU/mL vs 11.67 ± 6.63 mIU/mL], blood glucose [199.15 ± 63.72 mg/dL vs 96.6 ± 24.3 mg/dL], and testosterone [0.91 ± 0.2 nmol/L vs 0.53 ± 0.16 nmol/L] thereby triggering metabolic dysfunction and predisposed to lifestyle disorder. Also, the SNPs were found to be in linkage equilibrium and contributed to the development of PCOS differentially.

Impact of Pseudomonas putida RRF3 on the root transcriptome of rice plants: Insights into defense response, secondary metabolism and root exudation.

Pseudomonas putida is widely used as a biocontrol agent, however, mechanisms by which it initiates the plants' defense response remains obscure. To gain an insight into the molecular changes that occur in plants upon plant growth-promoting rhizobacteria colonization, root transcriptome analysis by using a microarray was performed in rice using P. putida RRF3 (a rice rhizosphere isolate). Data analysis revealed a differential regulation of 61 transcripts (48 h post-treatment), of which, majority corresponded to defense response, cell wall modification and secondary metabolism. Seven genes encoding salicylic acid (SA) responsive pathogenesis-related proteins were up-regulated significantly (fold change ranges from 1 to 4), which suggests that RRF3 has a profound impact on a SA-mediated defense signaling mechanism in rice. Investigations performed at later stages of RRF3 colonization by real-time polymerase chain reaction and high-performance liquid chromatography (HPLC) analysis confirmed the above results, demonstrating RRF3 as a potent biocontrol agent. Further, the impact of RRF3 colonization on root exudation, in particular, exudation of SA was investigated by HPLC. However, analysis revealed RRF3 to have a negative impact on root exudation of SA. Overall, this study shows that P. putida RRF3 immunizes the rice plants by re-organizing the root transcriptome to stimulate plant defense responses ('priming'), and simultaneously protects itself from the primed plants by altering the rhizosphere chemical constituents.

Green synthesized nickel nanoparticles for targeted detection and killing of S. typhimurium.

Simple and sensitive colorimetric immunosensor based on peroxidase mimetic activity and photothermal effect of nickel oxide nanoparticle (NiOGs) has been developed to detect and kill food borne pathogen Salmonella typhimurium. NiOGs showed superior peroxidase mimetic activity for oxidation of peroxidase substrate 3, 3', 5, 5'-tetramethylbenzidine (TMB). Oxidation of TMB by NiOGs followed Michaelis-Menten kinetics with Km and Vmax values of 0.25mM and 2.64×10-8M/s respectively. NiOGs was coated with citric acid (CA-NiOGs) followed by conjugation with antibody (anti-S. typhimurium) (Ab-CA-NiOGs) that effectively captured S. typhimurium. Colorimetric detection of S. typhimurium by Ab-CA-NiOGs showed a linear relationship between pathogen concentration (1×101 to 1×106cfu/mL) and color signal (652nm) with limit of detection (LOD) of 10cfu/mL. The proposed method showed no cross reactivity against other pathogens. Recovery of S. typhimurium in milk and juice samples was found to be 95 to 100% and 92 to 99% respectively. NiOGs exposed to laser irradiation showed dose dependent increase in temperature and singlet oxygen within 5min. Bacteria bound to Ab-CA-NiOGs after laser irradiation, induced membrane damage and reduced bacterial viability to 6%. The bifunctional peroxidase-mimetic activity and photothermal effect of NiOGs can be exploited in selective sensing and killing of target pathogens respectively in food products.