Hyperglycemia-induced miR182-5p drives glycolytic and angiogenic response in Proliferative Diabetic Retinopathy and RPE cells via depleting FoxO1.

PURPOSE: Diabetic Retinopathy (DR) is associated with metabolic dysfunction in cells such as retinal pigmented epithelium (RPE). Small molecular weight microRNAs can simultaneously regulate multiple gene products thus having pivotal roles in disease pathogenesis. Since miR182-5p is involved in regulating glycolysis and angiogenesis, two pathologic processes of DR, we investigated its status in DR eyes and in high glucose model in vitro. METHOD: ology: Total RNA was extracted from vitreous humor of PDR (n = 48) and macular hole (n = 22) subjects followed by quantification of miR182-5p and its target genes. ARPE-19 cells, cultured in DMEM under differential glucose conditions (5 mM and 25 mM) were used for metabolic and biochemical assays. Cells were transfected with miRNA182 mimic or antagomir to evaluate the gain and loss of function effects. RESULTS: PDR patient eyes had high levels of miR182-5p levels (p < 0.05). RPE cells under high glucose stress elevated miR182-5p expression with altered glycolytic pathway drivers such as HK2, PFKP and PKM2 over extended durations. Additionally, RPE cells under high glucose conditions exhibited reduced FoxO1 and enhanced Akt activation. RPE cells transfected with miR182-5p mimic phenocopied the enhanced basal and compensatory glycolytic rates observed under high glucose conditions with increased VEGF secretion. Conversely, inhibiting miR182-5p reduced Akt activation, glycolytic pathway proteins, and VEGF while stabilizing FoxO1. CONCLUSION: Glycolysis-associated proteins downstream of the FoxO1-Akt axis were regulated by miR182-5p. Further, miR182-5p increased expression of VEGFR2 and VEGF levels, likely via inhibition of ZNF24. Thus, the FoxO1-Akt-glycolysis/VEGF pathway driving metabolic dysfunction with concurrent angiogenic signaling in PDR may be potentially targeted for treatment via miR182-5p modulation.

In Silico Analysis of Functional SNPs in Genes of Complete Androgen Insensitivity Syndrome (CAIS): A Retrospective, Case-Control Study.

Background: Complete androgen insensitivity syndrome (CAIS) is one of the categories of androgen insensitivity syndrome (AIS) described as complete failure of the cell to react to androgens with external genitalia of a normal female. People with AIS condition are genetically male, with XY karyotype in each cell, but their bodies are unable to respond to male sex hormones (called androgens). It is associated with infertility as well as developing cancerous conditions. The genetic association of CAIS involves polymorphism of genes such as NR5A1, SOX9, SRD5A2, CBX2, GATA4, and SRY. Their mutation and participation in genetics of CAIS can be studied by Single Nucleotide polymorphism (SNP) analysis which is a way to detect genetic variations. SNP in coding region leads to synonymous and non-synonymous mutations. Hence, this study highlights analysis of SNPs associated with CAIS. Our aim is to study SNP analysis of NR5A1, SOX9, SRD5A2, CBX2, GATA4, SRY genes in Complete Androgen Insensitivity Syndrome. Methods: SIFT and Polyphen analysis was performed for all the genes and samples were subjected for PCR-SSCP technique. Results: SNPs were analyzed for the genes associated with CAIS. Benign and damaging SNPs were identified. DNA Samples were amplified using PCR technique and they will be analyzed using Single-strand conformation polymorphism (SSCP). Conclusions: As SNPs have decreased stability, damaging and benign character, they can be used as candidate hallmarks in study of Complete Androgen Insensitivity Syndrome.

Infertility and Social Issue Have the Most Significant Impact on Health-related Quality of Life among Polycystic Ovarian Syndrome Women in South India.

Background: Infertility can have a significant impact on the identity of women. Individual women, who are infertile, experience tragic emotions, as well as those who are sad for great losses, like the death of a loved one. In this case, the woman is experiencing the loss of the ability to procreate. Aim: In the present study, our major concern was to implement the health-related quality of life (HRQOL) Questionnaire on South Indian polycystic ovarian syndrome (PCOS) women to assess the impact of various clinical features of polycystic ovary syndrome on the HRQOL of South Indian women diagnosed. Settings and Design: A total of 126 females in the first phase and 356 females in the second phase between the age group of 18-40 years characterised under the Rotterdam criteria were selected for the study. Materials and Methods: The study was carried out in three different phases which included a one-to-one interview, group discussion and questionnaire session. In our study, we found that all the females who attend the study showed positivity for all the domains developed in the previous study and suggested that further domain can be developed. Statistical Analysis Used: Suitable statistical methods were used with Graph pad PRISM (version 6). Results: Hence, in our study, we developed a further new sixth domain called as 'social impact domain'. Among South Indian PCOS women, we found that infertility and social issue have the most significant impact on HRQOL. Conclusion: The revised questionnaire by including the sixth domain called 'Social issue' is likely to be useful in measuring the quality of health of female having PCOS in regard to South Indian population.

Screening of Polymorphisms of Transcription Factor 7-like 2 Gene in Polycystic Ovary Syndrome using Polymerase Chain Reaction-restriction Fragment Length Polymorphism Analysis.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common endocrine disorder occurring in premenopausal women, with a prevalence rate of 5%-7%. It has been observed in multiple number of studies the coexistence between diabetes mellitus 2 and obesity with this endocrinopathic disorder. Transcription factor 7-like 2 (TCF7L2) gene is shown to be associated with insulin secretion. AIM: To screen whether the gene variant of TCF7L2 (formerly TCF4) gene is significantly associated and has susceptibilities with type 2 diabetes in PCOS. This study is essential to uncover diabetogenic association of the TCF7L2 gene variants with PCOS. DESIGN: This was a hospital-based study. METHODS: In this work, blood samples from 43 PCOS patients with age and sex similar to 43 control samples were collected, followed by isolation of DNA. Further genotyping of the TCF7L2 gene was carried out by performing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). STATISTICAL ANALYSIS: Genotype frequencies of the TCF7L2 rs7903146 gene were checked by Hardy-Weinberg equilibrium of genotype in both PCOS and the control group, and also, the frequencies of the genotype were performed accordingly. RESULTS: There was no significant allelic variation observed among the patient and the control samples. From the patient details, it was observed that women between the age group of 21 and 25 years are susceptible to PCOS. CONCLUSION: From the PCR-RFLP analysis, it can be stated that there are no expected gene polymorphisms seen in this study, unlike the study carried out on the Chinese population where they observed genotype variations CC, CT, and TT. From this study, we can conclude that TCF7L2 rs7903146 gene cannot be considered as the candidate gene for the occurrence of PCOS.