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Introduction: Several studies assessed the sensitivity and specificity of plasma concentrations of hormones such as FSH and AMH as biomarkers that can predict the presence of sperm cells, prior to testicular sperm extraction (TESE) in azoospermic men. However, the literature contains conflicting reports in this context. Objective: To evaluate the potential of hormonal profile as a non-invasive predictor of mature sperm cells' presence in the testis of azoospermic men, and to assess if the source of these hormones- testicular or peripheral blood makes any difference. Methods: Clinical data, hormonal profile, and histological classification of 57 azoospermic men were analyzed. They were categorized into 4 following groups by their testicular pathologies: obstructive azoospermia, hypospermatogenesis, spermatocyte maturation arrest, and Sertoli cells only. The level of hormones was measured by the enzyme-linked immunosorbent assay. Statistical analyses tested the predictability of hormone values for sperm retrieval. Results: Overall, 28 patients (49%) had spermatozoa detected. FSH and Inhibin B measurements were statistically significantly different between the 4 groups with different testicular pathologies (p<0.001). AMH, FSH and Inhibin B were found to be independent predictor for positive sperm retrieval (p<0.05). However, a decision tree with random forest classifying the predictive ability of the variables to indicate the presence of sperm produced low accuracy. There was no significant difference between the hormone levels of peripheral blood source and testicular blood source hormone levels. Conclusions: AMH, FSH, and Inhibin B are optional markers for positive sperm retrieval in TESE. Increasing the research group may contribute to their ability to predict the presence of mature sperm cells in the testis of azoospermic men. Disclosure: No. [ABSTRACT FROM AUTHOR]

Introduction: Several studies assessed the sensitivity and specificity of plasma concentrations of hormones such as FSH and AMH as biomarkers that can predict the presence of sperm cells, prior to testicular sperm extraction (TESE) in azoospermic men. However, the literature contains conflicting reports in this context. Objective: To evaluate the potential of hormonal profile as a non-invasive predictor of mature sperm cells' presence in the testis of azoospermic men, and to assess if the source of these hormones- testicular or peripheral blood makes any difference. Methods: Clinical data, hormonal profile, and histological classification of 57 azoospermic men were analyzed. They were categorized into 4 following groups by their testicular pathologies: obstructive azoospermia, hypospermatogenesis, spermatocyte maturation arrest, and Sertoli cells only. The level of hormones was measured by the enzyme-linked immunosorbent assay. Statistical analyses tested the predictability of hormone values for sperm retrieval. Results: Overall, 28 patients (49%) had spermatozoa detected. FSH and Inhibin B measurements were statistically significantly different between the 4 groups with different testicular pathologies (p<0.001). AMH, FSH and Inhibin B were found to be independent predictor for positive sperm retrieval (p<0.05). However, a decision tree with random forest classifying the predictive ability of the variables to indicate the presence of sperm produced low accuracy. There was no significant difference between the hormone levels of peripheral blood source and testicular blood source hormone levels. Conclusions: AMH, FSH, and Inhibin B are optional markers for positive sperm retrieval in TESE. Increasing the research group may contribute to their ability to predict the presence of mature sperm cells in the testis of azoospermic men. Disclosure: No. [ABSTRACT FROM AUTHOR]

Introduction: In the present era, the extensive use of modern cell phones has raised the health risks associated with the radiofrequency-electromagnetic radiation (RF-EMR) they emit, which the human body can absorb. RF-EMR exposure can potentially harm the process of spermatogenesis by increasing oxidative stress, elevating mitochondrial reactive oxygen species (ROS), and triggering the formation of free radicals that leads to sperm DNA damage. Our previous research revealed decreased sperm motility and viability in semen samples exposed to RF-EMR through WiFi calling. Objective: To investigate the effects of WiFi radiation on genetic variants in human sperm using current-generation smartphones. Methods: Six healthy, normozoospermic men aged 25-35 provided semen samples for this study. A current generation smartphone was used in WiFi calling mode, specifically through Whatsapp voice call, to serve as the source of RF-EMR. The call duration was six hours. Semen samples were analyzed based on the 2010 WHO guidelines before and after the radiation exposure. The genomics facility at our university extracted DNA from semen samples of each participant before and after RF-EMR exposure to perform whole exome sequencing with alignment to the reference genome. After sequencing, we compared genetic variants for each participant before and after RF-EMR exposure. Results: After six hours of exposure, a significant decrease in sperm progressive motility was observed (p<0.05). Following exposure, the median number of exonic, nonsynonymous variants absent before exposure increased to 4 (IQR 3). Across the four successfully sequenced samples, we found 20 new, exonic, nonsynonymous variants after RF-EMR exposure. Significantly, male reproductive tissues expressed 12 variants, five associated with ciliary function and two related to mitochondrial function (see Table 1). Gene Chromosome Variant Location Function SPDYE11 Chr7 A > C Spermatids • Acetylcholine receptor activity • Cell cycle regulation SPDYE17 Chr7 A > C Spermatids • Protein kinase activity • Cell cycle regulation PRB2 Chr12 T > G salivary gland • Cell cycle regulation GGT2 Chr22 G > A Spermatids • Cell cycle regulation RGPD3 Chr2 C > T Spermatids • Ciliary function EPPK1 Chr8 G > A Spermatids, spermatocytes • Epithelial structure GOLGA6L4 Chr15 G > A Spermatids, spermatocytes • Ciliary function TCP11X2 ChrX T > C Spermatids • Acrosome reaction • Cilia function • Spermatid development HRNR Chr1 A > G Mitochondria • Metal ion binding POUF1 Chr13 > GCC Mitochondria • Structure & function of spermatozoa NPIPB13 Chr16 T > A Spermatids • Ciliary function • Sperm motility NBPF11 Chr1 A > C Spermatids, Sertoli cells, Leydig cells • Cell cycle regulation • Microtubule development ANKRD20A1 Chr9 T > A Spermatocytes • Ciliary function Table 1. Candidate genes with variants after six hours of phone radiation exposure Conclusions: Based on our pilot study, we observed a decrease in sperm progressive motility and an increase in potentially related genetic variants associated with sperm motility following six hours of exposure to RF-EMR. These preliminary results highlight the need for further research to investigate the effects of RF-EMR exposure on semen parameters and to determine the clinical significance of these genetic variants. Disclosure: No. [ABSTRACT FROM AUTHOR]

Introduction: The testicular microenvironment, consisting of cells including Sertoli and peritubular cells, releases paracrine factors that may regulate testicular function. In previous research, Leptin was identified as a critical paracrine factor in the testicular microenvironment. Leptin is secreted by Sertoli and peritubular cells and binds to its receptor in Leydig cells. This interaction triggers the expression of the desert hedgehog signaling pathway, which is essential for Leydig stem cell differentiation into adult Leydig cells, which are capable of producing testosterone. However, our earlier study had a limitation: the human cell model used was derived from testes biopsies, lacking an intact immune microenvironment. This study investigates the effect of Leptin on a complete testicular immune microenvironment in a murine model. Objective: To investigate the impact of Leptin on a complete testicular immune microenvironment. Methods: We used 6-week-old C57BL6 mice, exposed to two different concentrations of Leptin, 10 and 100 mg/ml, for seven days. After the treatment period, the animals were humanely euthanized, and their blood and testes were collected. Blood samples underwent a complete blood count differential profile analysis using a Hemavet Mascot Multispecies Hematology System Counter 1500R. Testes samples were subjected to comprehensive immunophenotyping, allowing us to assess the impact of Leptin treatment on various immune cell types within the testes Results: Leptin at a low dose (10 mg/ml) showed increased, but not significant, levels of serum testosterone (13.20 + 0.37 ng/dl), while LH and FSH levels significantly (p<0.05) increased. In contrast, animals receiving 100 mg/ml Leptin showed no significant changes in hormone levels. CBC profiling indicated that lymphocytes and platelets were significantly influenced, specifically with low doses of Leptin. Comprehensive immunophenotyping revealed a significant increase in CD4+ (helper T cells) and CD8+ (cytotoxic T lymphocytes) cell numbers in animals treated with low doses of Leptin, a pattern not observed with high doses. Conclusions: This study uncovers previously unexplored aspects of how paracrine factors, released by Sertoli and peritubular myoid cells, can modulate the immune microenvironment within the testis, potentially impacting testosterone production. Future research will investigate the relationship between changes in immune cell types, increased T cell numbers, and their influence on Leydig stem cell differentiation into adult Leydig cells Disclosure: No. [ABSTRACT FROM AUTHOR]