The somatic gonad plays a central role in supporting germline development and maintaining oocyte and embryonic integrity, both of which are essential for sustained reproductive function. With age, deterioration of somatic gonadal structures compromises germline support, leading to declines in oocyte morphology and increased embryonic abnormalities. The insulin/IGF-1 signaling (IIS) pathway regulates longevity and reproductive aging, and its reduction (rIIS) in the daf-2(e1370) mutant extends reproductive span in Caenorhabditis elegans. While prior work has largely focused on germline stem cells and oocyte maturation, the molecular mechanisms by which IIS preserves somatic gonadal architecture and thereby maintains oocyte and embryonic morphological integrity remain poorly understood. Elucidating how reduced IIS influences the somatic gonad can provide insights into the coordination of systemic aging pathways with tissue-specific mechanisms that sustain reproductive health. We identified somatic gonad–enriched genes from temporal transcriptomic datasets of reduced IIS animals, highlighting collagens consistently upregulated with age. Using RNAi knockdowns in both systemic and somatic gonad–specific RNAi-sensitive strains of Caenorhabditis elegans, we examined effects on self-reproductive span, oocyte and embryonic morphology, and somatic gonad integrity. Age-associated morphological changes were assessed by differential interference contrast microscopy, and regulatory dependencies were tested via quantitative polymerase chain reaction following knockdown of key transcription factors. Several collagens were found to be essential for maintaining somatic gonadal architecture and preserving oocyte morphology during aging. Gonad-specific knockdown of these collagens caused structural deterioration, impaired oocyte and embryo quality, and accelerated reproductive decline. These defects occurred without a major reduction in total progeny number, suggesting that age-related reproductive loss arises primarily from tissue deterioration rather than immediate fertility impairment. Expression and function of these collagens were largely dependent on key transcription factors downstream of IIS. Our findings uncover a previously unrecognized role for somatic gonadal collagens in preserving reproductive function under IIS. By maintaining gonadal architecture and sustaining oocyte and embryonic morphological integrity, these collagens act as downstream effectors of IIS-mediated reproductive longevity. Targeting gonadal collagen networks may therefore represent a potential strategy to mitigate age-related reproductive decline and warrants further investigation in higher-order models. Healthy fertility depends not just on eggs, but also on the cells that surround and support them. These supporting structures, known as the somatic gonad, play a vital role in keeping eggs healthy during aging. In this study, we explored how a protein called collagen—best known for providing structure in skin and other tissues—helps maintain the health of reproductive organs in aging animals. Using the model organism Caenorhabditis elegans, we focused on how reduced insulin/IGF-1 signaling (IIS)—a condition known to extend lifespan—affects the function of collagen genes in reproductive tissues. We found that certain collagen genes are especially active in the somatic gonad under reduced IIS conditions. When we experimentally lowered the activity of these collagen genes, the structure of the reproductive organs broke down faster, and egg quality declined more rapidly—even though total offspring numbers were not immediately affected. Our findings suggest that collagen is essential for preserving the structural health of reproductive organs as animals age, and that this process is regulated by the IIS pathway. Targeting collagen pathways might offer new strategies to support fertility and reproductive longevity.

Background. The clinical utility of procalcitonin (PCT) and proadrenomedullin (ProADM) in children with malignancies remains inadequately characterized. This study was designed to assess baseline PCT and ProADM levels at the time of leukemia diagnosis and to compare their profiles during subsequent episodes of febrile neutropenia (FN). Methods. Children aged 18 years or younger with newly diagnosed acute leukemia, including acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML), were prospectively recruited for this study. Serum levels of PCT and ProADM were measured at baseline, prior to the initiation of chemotherapy at the time of diagnosis, and were reassessed during subsequent episodes of FN. Results. A total of 80 children with acute leukemia were prospectively enrolled, with a median age of 5 years (interquartile range [IQR]: 3-7 years). The study population comprised of ALL in 67.5% of cases and AML in 32.5%. At the time of diagnosis, prior to chemotherapy initiation (n=80), the median serum PCT level was 0.19 ng/mL (IQR: 0.07–0.54), while the median ProADM level was 0.04 nmol/L (IQR: 0.01–0.07). Among the 80 patients, 32 children subsequently developed FN and had paired serum samples available for comparative analysis. During FN episodes, both biomarkers showed a significant increase relative to baseline values. Median PCT increased from 0.16 ng/mL (0.08–0.52) at diagnosis to 0.32 ng/mL (0.08–0.50) during FN (P = 0.03), while median ProADM increased from 0.03 nmol/L (0.006–0.05) to 0.41 nmol/L (0.20–0.81) (P < 0.001). At the time of leukemia diagnosis, splenomegaly was the only clinical factor significantly associated with elevated baseline PCT levels (P = 0.010). Subgroup analysis revealed distinct biomarker patterns: in children with ALL, PCT levels remained largely unchanged between diagnosis and FN, possibly reflecting an underlying baseline inflammatory state, whereas ProADM showed a significant rise during FN, suggesting greater specificity for infectious events. In contrast, in AML, both PCT and ProADM increased significantly during FN, indicating their potential utility as infection-related biomarkers in this subgroup. Conclusion. Both PCT and ProADM were significantly elevated in FN when patients with acute leukemia were analyzed as a whole. However, subgroup analysis revealed differing patterns: in ALL, only ProADM remained significantly associated with FN, whereas PCT showed no significant association. In contrast, in AML, both biomarkers were significantly elevated. These findings suggest that ProADM may have greater clinical utility in guiding the management of febrile episodes, particularly in ALL.

Background: Polycystic ovary syndrome (PCOS) is a complex endocrine disorder with a multifactorial aetiology, including genetic and epigenetic components. Despite increasing evidence highlighting the role of epigenetic modifications in PCOS, their contribution to disease progression remains unclear. Aim: This study aimed to investigate the role of DNA methylation in the aetiology of PCOS. Settings and Design: This is a hospital-based observational study. Materials and Methods: The materials and methods of this study were to examine the role of DNA methylation in PCOS. Genomic DNA was isolated using the QIAamp DNA Mini Kit, and DNA methylation profiling was performed using the Infinium Human Methylation 850K (EPIC) array in 43 PCOS cases compared with controls. Statistical Analysis Used: The Student’s t-test (two-tailed) was used to compare the means and derive statistical significance from the independent groups. The ChAMP software package was used for methylation analysis on the R interface v4.2.0. Results: Differential methylation analysis identified 243 differentially methylated CpG sites within 26 differentially methylated regions (DMRs) (|Δβ| > 0.1, P < 0.05). Notably, most DMRs were located in promoter regions near CpG islands and shores, with a significant proportion (23.07%) mapping to chromosome 6. The majority (76.9%) of DMRs exhibited hypomethylation, whereas the top two DMRs were hypermethylated. Functional enrichment analysis revealed that differentially methylated genes were associated with key pathways, including Th17 cell differentiation, steroid biosynthesis and autoimmune thyroid disease. Gene ontology analysis highlighted enrichment in immune-related biological processes, MHC protein complexes and molecular functions, including prenyltransferase and transaminase activities. Conclusion: Our findings underscore the potential contribution of epigenetic modifications to PCOS pathogenesis, particularly through immune regulation and steroid metabolism. Further studies with larger cohorts and functional validation are warranted to establish the role of these epigenetic changes in PCOS.

Background: Heavy metals, such as Cadmium (Cd), Arsenic (As), Mercury (Hg) and Lead (Pb) might potentially induce reproductive toxicity in male infertility patients, regardless of the varying concentrations of these heavy metals in the blood. Genetic polymorphism is one of the least studied internal contributing factors in male infertility cases associated with high level of heavy metal in blood. Therefore, this study aims at identifying the difference in the serum levels of heavy metals in non-obstructive azoospermia (NOA) patients associated with genetic variants. Methods: It’s a hospital based observational study where patients reporting with azoospermia due to hypospermatogenesis (HS) were recruited prospectively. Comprehensive clinical history, and blood samples were collected. Whole exome sequencing (WES) and was performed for 50 HS patients to identify variants. Inductively coupled plasma mass spectrometry (ICP-MS) was performed to assess levels of Cd, As, Hg and Pb levels in serum samples of 50 HS patients. Statistical analysis was performed to determine difference in heavy metal concentration of HS patients with and without the presence of metallothionein gene associated single nucleotide polymorphism (SNP). Results: Genomic analysis for SNPs identified deleterious candidate variants in MT1A (rs11640851 and rs8052394) associated with 18/50, MT1E (rs138690474) associated with 4/50 and MT4 (rs11643815) associated with 5/50 HS patients. A statistically significant difference in the blood concentration of Cd and Hg was observed in HS patients associated with metallothionein gene SNPs. Conclusions: This exploratory genomic analysis conducted on HS patients shows prevalence of deleterious candidate SNPs in metallothionein gene. The HS patients with candidate SNPs showed higher levels of Cd and Hg which indicate the genomic susceptibly towards heavy metal-induced reproductive toxicity.

Primary objective was to compare the performance of proadrenomedullin (ProADM) and procalcitonin (PCT) for diagnosing clinically documented infections (CDI)/microbiologically documented infections (MDI)/fever without focus (NF) in children with cancer having febrile neutropenia (FN). The secondary objective was to compare the prognostic utility of PCT and ProADM for identifying adverse clinical outcome. Cancer patients (aged ≤ 18 years) presenting with FN were included; those who had received antibiotics within the last 14 days were excluded. Serum PCT and ProADM were estimated on days 1, 3 and 7 of enrolment. Adverse outcome was defined in terms of clinical non-response and day 30 mortality. We recruited 345 children. PCT in children with MDI were significantly higher than those with CDI on day 1 (P = 0.031) and day 7 (P < 0.001); PCT ≥ 0.21 ng/mL on day 1 had a 77

Do mitochondrial variants play a role in the pathogenesis of Premature Ovarian Insufficiency (POI)? Genetic variants in mitochondrial and fatty acid oxidation pathways are key contributors to the pathogenesis of POI, underscoring its intricate and multifactorial etiology. Mitochondria are essential for energy production in oocytes, facilitating folliculogenesis and oocyte maturation by generating ATP through the β-oxidation of fatty acids via the mitochondrial fatty acid shuttle (MFAS). This pathway, regulated by key enzymes such as CPT1, CPT2, CACT, OCTN2, and L-carnitine, is essential for spindle assembly, chromosomal segregation, and meiotic progression-factors critical for oocyte competence and embryo development. Disruptions in MFAS can impair β-oxidation and ATP production, thereby compromising follicular and oocyte health. Despite its fundamental role, the contribution of MFAS dysfunction to the pathophysiology of Premature Ovarian Insufficiency (POI) remains unexplored. Whole Exome Sequencing (WES) was conducted on blood DNA samples from idiopathic POI women (n = 10) to identify genetic variants. Serum reproductive hormone levels (E2, FSH, AMH, PRL) were measured, followed by in-silico analyses and protein modeling to assess the impact on protein structure/function. C57BL/6 adolescent mice (3–5 weeks, n = 4/group) were treated with MFAS inhibitors. Histological analysis of ovaries and other metabolic organs was performed post-treatment. The study lasted for 2 years. WES was performed on POI women, with data analyzed using our custom pipeline and in-silico tools (CADD, SIFT, PP2, CONDEL, FATHM) for predicting the impact of mitochondrial variants on protein function and structure, utilizing protein modeling tools like Pymol and ProteinPaint. Mice were treated with MFAS inhibitors (Etomoxir, Omeprazole, L-Amino Carnitine, Etoposide, Mildronate), along with cyclophosphamide (positive control) and saline (negative control), followed by collection of organs (ovaries, kidneys, liver, spleen) for H&E analysis. A novel variant in OCTN2 (SLC25A5) and variants of uncertain significance (VOUS) in CPT1A, CPT1B, and CPT2 were identified within genes related to the mitochondrial fatty acid shuttle (MFAS). In terms of mitochondrial function, two novel pathogenic variants (ARHGEF28 and AOX1) along with six likely pathogenic variants (MT-ND2, MT-ATP6, MT-CYB, MT-ND4, MT-ND5, MT-CO3) were discovered. Notably, CPT1, CPT2, SLC25A5, AOX1, and ARHGEF28 exhibited high predicted local distance difference test (pLDDT) scores, suggesting significant structural alterations.SLC25A5 and AOX1 displayed very high pLDDT scores (>90), while ARHGEF28 scored between 70 and 90.Functionally, CPT1, CPT2, and SLC25A5 contribute to ATP production, AOX1 regulates ROS generation, and ARHGEF28 affects cell cycle progression. Furthermore, 18 pathogenic variants and 29 VOUS were found in mitochondrial and cell-cycle genes, many of which regulate meiosis, follicular development, and ovulation, indicating their potential impact on fertility. In POI mouse models, MFAS inhibition and a positive control led to ovarian shrinkage, follicular atresia, and systemic organ abnormalities (kidney, liver, spleen), resembling cyclophosphamide POI-induced histological changes. These findings highlight the impact of MFAS dysfunction on ovarian folliculogenesis and systemic health, providing new insights into the mitochondrial contribution to Premature Ovarian Insufficiency (POI). Small sample size of human participants (n = 10), which may limit the generalizability of the findings, as well as the inability to obtain human ovarian tissue due to ethical constraints. Additionally, the lack of long-term treatment analysis restricts our understanding of the chronic effects of MFAS inhibition. Exploring mitochondrial dysfunction alongside existing diagnostic criteria could facilitate early detection of POI and offer multiple avenues for early disease diagnosis. Integrating this approach with established hormonal diagnostic methods may provide more comprehensive insights, provided that similar studies are conducted in larger cohorts. No

The most prevalent reproductive endocrine condition affecting women of reproductive age is called polycystic ovary syndrome, or PCOS. Hyperandrogenism, irregular ovulation, and polycystic ovarian shape are the hallmarks of PCOS. Despite the fact that PCOS has been recognized for more than a century, there is ongoing debate over its diagnosis, origin, clinical characteristics, and course of therapy. The first scientific study of (PCOS) was conducted in 1935 by Stein and Leventhal, although Vallisneri provided the first account of PCOS in 1721, marking the historical evolution of the syndrome. The first scientific PCOS diagnostic criteria were proposed in 1990 at an NIH-sponsored conference. The syndrome is most often known as Polycystic Ovary Syndrome (PCOS), and the NIH 2012 criteria are now the mainstream diagnostic standards used, i.e., Rotterdam 2003 criteria with phenotypic classification. The last guideline on PCOS, i.e., the international evidence-based policy (2018), amended in 2023, is acceptable to all groups working on PCOS. AMH may be the most effective biomarker currently identified for PCOS. Complex and heterogeneous, PCOS is influenced by environmental and epigenetic factors in addition to genetic vulnerability. This chapter on PCOS aims to provide an introductory note along with the historical evolution of PCOS.

IntroductionHypospermatogenesis is a common histopathological subtype of non-obstructive azoospermia and is characterized by a decrease in the total number of germ cells within the seminiferous tubule as a result of spermatogenic failure. Determination of genetic factors before intracytoplasmic sperm injection can prevent the inheritance of these factors, as hypospermatogenesis patients gives high successful sperm retrieval rate. This study aimed to identify the structural variants associated with idiopathic hypospermatogenesis (iHS) by analyzing patient cohorts diagnosed with azoospermia using whole exome sequencing.MethodsIt is a hospital-based observational study in which patients reporting with azoospermia due to spermatogenic failure were recruited prospectively. Comprehensive clinical history, blood samples, semen analysis parameters, and reproductive endocrine evaluation reports of 51 hypospermatogenesis patients were collected. The known genetic causes were investigated using XY fluorescent in situ hybridization and Yq microdeletion for exclusion. Whole exome sequencing was performed, and the data of 42 iHS patients was analyzed to identify single nucleotide variants associated with diagnostically important male infertility genes.ResultsGenomic analysis of SNVs identified rare deleterious candidate variants in CFTR (c.1265C>T; p.Ser422Phe), CYP21A2 (c.955C>T; p.Gln319Glu), SRD5A2 (c.737G>A; p.Arg245Gln), LHCGR (c.378A>C; p.Lys126Asn) and AR (c.2179C>A; p.Arg727Ser) genes associated with 7/42 idiopathic hypospermatogenesis patients. In silico analysis of variants shows deleterious and probably damaging effects on canonical transcripts of the genes.DiscussionThis exploratory genomic analysis conducted on idiopathic hypospermatogenesis patients shows prevalence of rare deleterious candidate variants in genes associated with human male infertility. The candidate variants in idiopathic hypospermatogenesis patients are heterozygous and genotypically associated with syndromic male infertility. The symptomatic heterozygosity leading to mild spermatogenic failure resulting in hypospermatogenesis points towards a multifactorial etiology of the disease. This study justifies the importance of genetic screening of idiopathic hypospermatogenesis patients for the presence of structural variants in known human male infertility genes.

Objectives Hypospermatogenesis (HS) is a common histopathological subtype of non-obstructive azoospermia (NOA), characterized by a decrease in the total number of germ cells within the seminiferous tubule. The diagnosis of HS is made by invasive procedures like testicular biopsy or fine needle aspiration (FNA). MicroRNAs (miRs) are biomolecules with emerging roles as diagnostic biomarkers for diseases. This study aimed to investigate the differential miR expression profile in idiopathic HS (iHS) to explore candidate spermatogenic miRs in human male infertility. Material and Methods In this observational study, patients reporting azoospermia due to spermatogenic failure were recruited prospectively. Genetic exclusion was performed using XY-fluorescent in-situ hybridization (XY-FISH) and Yq microdeletion. Testicular FNA samples were used for total ribonucleic ccid (RNA) isolation. Small RNA sequencing-based exploratory analysis was performed on 20 iHS patients and five normospermatogenesis (NS) patients. nCounter miRNA expression based validation was performed for four iHS and four NS patients. Results Analysis of miRs in testicular tissue showed differential expression patterns having 49 downregulated and 3 upregulated miRs between iHS and NS patients, with miR-379-5p, miR-449a, miR-181c, miR-34b-3p, and miR-122b-5p being notable candidate spermatogenic miRNAs. Pathways such as Phosphatidylinositol 3 Kinase-Protein Kinase B (PI3K-Akt) and mitogen-activated protein kinase (MAPK) signaling and molecular functions like apoptosis and cell differentiation were significantly enriched in iHS patients. This is the first study of its kind to investigate the differential expression of micro-ribonucleic acid (miRs) in a cohort of NOA patients exclusively of the HS subtype. Comparing the study results to previously published data revealed that dysregulated spermatogenic miRs are shared in HS and other NOA subtypes. The analysis of spermatogenic miRs according to each patient’s profile showed significant dysregulation in miR expression, linked to 40% of cases with idiopathic HS. Conclusion This study provides important insight into the potential of miRNA to be used as a biomarker for the diagnosis of iHS. Although the study is based on a relatively low sample size, it provides a proof of concept which can be validated in a larger cohort.

Abstract:Endometriosis is a prevalent condition where tissue similar to the uterine lining grows outside the uterus, causing pain and infertility. Diagnosing endometriosis typically requires invasive procedures such as laparoscopy. MicroRNAs (miRNAs) have emerged as promising noninvasive biomarkers for various diseases, including endometriosis. However, studies have shown inconsistent miRNA expression patterns across populations. This study aims to validate circulating miRNAs as biomarkers for endometriosis in Indian women, addressing the limited validation data available for this population. This comprehensive review identified nine circulating miRNAs based on reproducibility and consistent expression patterns. Women with advanced-stage endometriosis (n = 12) and controls (n = 11) were recruited. Plasma samples were collected based on clinical symptoms, CA-125 levels, ultrasound, MRI findings, and laparoscopic confirmation. miRNA expression was quantified using qRT-PCR, and receiver operating characteristic (ROC) analysis was performed to assess diagnostic potential. Nine miRNAs (miR-451a, let-7b, miR-150-5p, miR-17-5p, miR-3613-5p, miR-20a-5p, miR-342-3p, miR-125b-5p, and miR-21-5p) were analyzed. Among them, miR-451a and miR-20a-5p exhibited significantly lower expression in endometriosis patients (n = 12) compared to controls (n = 11). ROC analysis demonstrated promising diagnostic potential for these miRNAs. miR-451a showed distinct trends compared to previous studies, while miR-20a-5p was consistent with earlier research. Although encouraging, these findings are based on a limited sample size. Larger multicenter studies across diverse populations using reliable reference genes are needed to fully assess the diagnostic value of these miRNAs as biomarkers for endometriosis. Lay summary:Endometriosis is a common condition where tissue similar to the uterine lining grows outside the uterus, causing pain and infertility. Diagnosing it usually requires invasive procedures such as laparoscopy. We focused on microRNAs (miRNAs), small molecules in plasma that could offer a noninvasive way to diagnose endometriosis. After reviewing 45 research articles, we identified 102 miRNAs that were elevated and 197 that were reduced in endometriosis patients. From these, we selected nine promising miRNAs for validation in the Indian population. We collected blood samples from 12 women with endometriosis and 11 healthy controls. Our analysis showed significant differences in miRNA expression, with miR-451a and miR-20a-5p showing strong potential to distinguish between endometriosis patients and healthy individuals. These findings suggest that miRNAs could improve the diagnosis of endometriosis in a less invasive manner. In conclusion, our research highlights the potential of miRNAs in advancing endometriosis diagnosis and management.

BACKGROUND:Age-related decline in reproductive function is a hallmark of organismal aging, yet the molecular mechanisms driving this process remain incompletely understood. The insulin/IGF-1 signaling (IIS) pathway is highly conserved and influences both lifespan and reproductive aging in Caenorhabditis elegans, where reduced IIS extends reproductive span. While prior studies have examined isolated tissues or time points, a comprehensive temporal analysis of gonadal transcriptional dynamics under reduced IIS has been lacking. Here, we compared IIS-dependent regulation of the gonadal transcriptome with that of other somatic tissues to uncover tissue-specific mechanisms of reproductive aging. METHODS:Bulk RNA sequencing was performed on distal gonads dissected above the spermatheca from wild-type N2 and daf-2(e1370) mutant animals, a well-established model of reduced IIS. Samples were collected at four physiologically relevant adult stages-Day 1 (young adult), Day 2, Day 6, and Day 10-covering early to late reproductive periods. In parallel, whole-worm RNA-seq was conducted for N2 and daf-2 at Day 1 and Day 10 to enable systemic comparisons. Differential gene expression analyses identified IIS-responsive transcripts that were either gonad-specific or non-gonadal. Expression datasets were further analyzed using self-organizing maps (SOMs) with hierarchical clustering. Gene network construction, functional enrichment, transcription factor enrichment, and conservation analyses were performed, and differential expression profiles were integrated with publicly available germline-, gamete-, and somatic tissue-enriched datasets. RESULTS:Temporal transcriptomic profiling revealed distinct IIS-dependent expression trajectories in gonadal versus non-gonadal datasets. SOM-based clustering resolved temporally regulated expression modules, while network and enrichment analyses uncovered a multilayered regulatory architecture within the gonad. Gonadal expression was enriched for structural, extracellular matrix, and signaling pathway genes, whereas non-gonadal data showed enrichment for stress response and longevity-associated pathways. Integration with germline-, gamete-, and somatic tissue-enriched datasets distinguished tissue-specific regulatory signatures. Importantly, IIS-regulated gonadal components included genes highly conserved with human orthologs. CONCLUSION:This study provides a high-resolution temporal map of the gonadal transcriptome under reduced IIS and highlights gene modules potentially critical for reproductive maintenance. These findings offer a resource for dissecting tissue-specific aging programs and insulin-dependent regulation of reproductive health.

Aim: Epigenetic alterations have been reported in patients with pituitary tumors and those on antipsychotic drugs, which are also responsible for hyperprolactinemia. This suggests a possible role of epigenetics in the etiopathology of hyperprolactinemia. Methods: The study recruited 83 hyperprolactinemia cases with prolactin > 100 ng/mL and 65 controls. Global DNA methylation status was studied by MethylFlash Methylated DNA Quantification Kit and genome-wide methylation analysis (GWMA) by Infinium Methylation EPIC BeadChip 850K array. Results: Hyperprolactinemia cases showed significant global DNA hypermethylation compared to controls. Around 66.67% of hypomethylated and 12.9% of hypermethylated cases were on antipsychotics. Gene enrichment analysis of 5-cytosine-phosphate-guanine-3 (CpG) site-associated genes demonstrated significantly enriched major histocompatibility complex (MHC)-related protein classes and cellular components. Conclusions: The study suggested the role of epigenetics in the etiopathology of hyperprolactinemia.

Objectives: The objectives of this study were to compare the sperm parameters and sperm phospholipase C zeta (PLC zeta) expression profile before and after vitrification. Materials and Methods: Pre-vitrification and post-vitrification analysis of semen samples of 14 infertile men was carried out for sperm parameters such as motility, vitality, and morphology by standard semen analysis procedures, whereas the proportion of sperms exhibiting PLC zeta and its localization pattern was assessed by indirect immunofluorescence. The temporal analysis was done thrice: over 1 week, 1 month, and 3 months of cryostorage. Statistical Analysis: Statistical analyses were performed using GraphPad Prism version 8.0.1 (San Diego, California, USA). Data were expressed as mean +/- standard deviation. Parametric or non-parametric tests were employed for comparisons according to the normality of data distribution and the available data set. P value of 0.05 was considered statistically significant. Results: Vitrification was found to be associated with a decrease in the percentage of sperm motility (P <= 0.0001), vitality (P <= 0.0001), spermatozoa exhibiting normal morphology (P > 0.05), and PLC zeta protein (P > 0.05), however, the latter two, only, insignificantly. There was increased dominance in the post-acrosomal localization of PLC zeta after vitrification (P <= 0.001). Conclusions: The post-acrosomal localization of PLC zeta has been reported to have the highest positive correlation with oocyte fertilization and the present study showed the predominant pattern of the same. The implications for quality maintenance for long storage periods can be suggested as better sperm quality was observed at 3 months of storage during this study. This raises the hypothesis that the vitrification method of sperm cryopreservation may be the method of choice for routine clinical use in the assisted reproductive technology settings.

PCOS is a common endocrinopathy among women of reproductive age, with a worldwide prevalence of 8 to 13%, depending on the criteria used for diagnosis. It is characterized by a constellation of features, including oligo/anovulation, clinical and/or biochemical hyperandrogenism, and polycystic ovarian morphology. PCOS is one of the common causes of female infertility. It is also associated with metabolic derangements, including obesity, insulin resistance, and compensatory hyperinsulinemia, which increase the likelihood of developing type 2 diabetes mellitus. Despite extensive research, the etiology of PCOS remains largely unknown. It seems likely that the hypothalamic-pituitary-ovarian axis dysfunction, partial folliculogenesis arrest, insulin resistance, and ovarian and adrenal androgen secretion may play a role in the pathogenesis of PCOS. Familial clustering of the cases of PCOS points to a genetic component linked with it. The initial genetic studies suggest an autosomal dominant pattern of inheritance of the disorder in some families; however, most studies support multifactorial origin. Since PCOS is a complex trait, the typical form of inheritance of PCOS follows a non-Mendelian pattern and involves complex genetic mechanisms. Studies involving linkage and association have suggested a connection between genetic variations and the risk of developing PCOS in certain families or populations. Through genome-wide association studies and next-generation sequencing techniques, several candidate genes have been identified that play a role in the etiopathogenesis of the disorder. Pathogenic variants of various genes such as INSR, IRS1, GHRL, LDLR, MC4R, ADIPOQ, UCP1, UCP2, UCP3, FTO, PCSK9, FBN3, NEIL2, FDFT1, PCSK9, CYP11, CYP17, CYP21, HSD17, STAR, POR, AKR1C3, AMH, AMHR2, INHBA, AR, SHBG, LHR, FSHR, FSH β, SRD5A, GATA4, THADA, YAP1, ERBB2, DENND1A, FEM1B, FDFT1, NEIL2, TCF7L2, etc. in some PCOS cases are linked as underlying etiologic associations. This review aims to provide insight into the current genetic knowledge about PCOS. Discovering the genetic factors and pathways involved in the disorder will help us better comprehend the underlying mechanisms of the disorder.

Telomeres are vital for cellular function. Oxidative stress is detrimental to telomere function as it causes telomere attrition. Increase in oxidative stress has a negative impact on sperm telomere and chromatin integrity and therefore, on sperm function and male fertility. TERRA, a long non-coding RNA is a fundamental component required for telomere length homeostasis. Aim: The present work aimed to study the effect of H2O2 induced oxidative stress on sperm TERRA expression and chromatin function. Methodology: 5 infertile and 5 fertile males were selected for the study. Baseline semen analysis was done as per WHO manual 2021. Sperm TERRA expression was assessed by real time qPCR. To assess chromatin function nuclear chromatin decondensation test was done. Results: Study showed that 500µM H2O2 increased TERRA expression as compared to untreated groups and the difference was found to be statistically significant. As compared to controls, cases showed more fold increase in TERRA expression. When treated samples of control and cases were compared, cases had more TERRA expression than controls however, the baseline TERRA expressions in untreated samples were less in cases as compared to controls. Both comparisons yielded statistically nonsignificant results. When nuclear chromatin decondensation was compared before and after H2O2 treatment, the percentage of sperm head decondensation decreased in both cases and controls. When treated and untreated samples of cases and controls were compared, the difference was statistically nonsignificant. Conclusion: Oxidative stress led to an increase in the expression of sperm TERRA and had a negative impact on nuclear chromatin decondensation. Similar studies with greater sample size can provide much more insight into TERRA mediated regulation of telomere function and chromatin integrity and how it is affected by oxidative stress.