Objectives: The underlying pathophysiological mechanisms of azoospermia is still unclear. The aim of the study was to evaluate nitric oxide synthase (NOS) isoforms and free radical release in testicular sperm extraction (TESE) in infertile men with azoospermia. Materials and methods: The study included 40 men (mean age: 37.2±2 years; range 25 to 55 years) with azoospermia which were divided into two groups: spermatozoa-present (n=20) and spermatozoa-absent (n=20). Testicular samples were examined morphologically, immunohistochemically, and biochemically. The TESE samples were examined according to number of mast cells stained with toluidine blue; immunohistochemically with three types of NOS isoforms, and free radicals were measured with chemiluminescence method, respectively. Results: Endothelial NOS (eNOS) reaction in spermatozoa-present group was considerably higher than spermatozoa-absent group (p<0.001). Compared to the spermatozoa-present group, inducible NOS (iNOS) reaction was higher than the spermatozoa-absent group (p<0.05). Neuronal NOS (nNOS) reaction was only prominent in Leydig cells in both groups. Mast cells increased (p<0.05) in the interstitial area surrounding seminiferous tubules in spermatozoa-absent samples. Superoxide radical generation in spermatozoa-present samples was significantly lower (p=0.0003). The peroxynitrite ratio in spermatozoa-absent samples was significantly higher (p=0.0038). Conclusion: These results suggest that eNOS, iNOS, and mast cells play an important role in spermatogenesis process in azoospermic men.

Backround: The aim of thisprospectivecohortstudywastoevaluatethedevelopmentalpotential of metaphase I (MI) oocytes that failed to reach maturation following rescue in vitro maturation (IVM). Method: ICSI was performed to these oocytes and they were subsequentlyplaced in the calcium ionophore-containing medium for 20 minutes, washed in culture medium and incubated until fertilization assessment.In total of 263 patients and 276 ICSI cycles are included.Of these, 28 patients had onlyimmature oocytes that did not respond to IVM. Results: The mean age of the study population was 36.4±4 and the primary indication for treatment was low ovarian reserve, accounting for 39.3% of cases.The average number of retrieved oocytes was 6.7, the average count of MII oocytes was 4, while the average count of MI oocytes was 2. Fertilization of MI oocytes occurred in 17 out of 28 patients(60.7%).Among the fertilized zygotes, 29.4% were classified as Z2PN, while 52.9% were identified as Z3PN.Amoung 8 embryos that reached day 4; 5 were fresh trasnferred. One patient had ß HCG positivity. 3 embryos are frozen and thawed transferred; one patient had positive ß HCG. The clinical pregnancy rate was determined to be 25% (2/8). Only one pregnancy ended in live birth (live birth rate 12.5%). Conclusion: This study demonstrated the developmental potential of MI oocytes that failed to mature after rescue IVM. Utilizing these MI oocytes could increase the number of transferable embryos, enhance the efficiency of ICSI cycles, and improve pregnancy rates, as each embryo holds clinical significance.

Purpose: To investigate the bacterial microbiome pattern of testicular tissue and its potential role in male infertility and in predicting micro‐TESE results. Methods: A total of 180 infertile men, including nonobstructive azoospermia (NOA) cases, referred to as the study group, and men with previous unsuccessful IVF treatments and/or a sperm DNA fragmentation index of 50% or greater. NOA cases, referred to as the study group, and men with previous unsuccessful IVF treatments and/or a sperm DNA fragmentation index of 50% or greater, referred to as the control group, were enrolled in the study. The study group was also divided into micro‐TESE (−) and micro‐TESE (+) subgroups. Testicular tissue microbiome analysis was performed, and the results were compared between groups and subgroups of the study group. Histological changes and ZO‐1 protein expression patterns were also compared. Results: Microbial composition was determined only in NOA patients. Although the composition of the microbiota was similar, micro‐TESE (−) patients showed differences in phylum and genus diversity with increased abundance of the phyla Bacteroidota and Actinobacteriota, and the genera Prevotella and Bifidobacterium in one of the two subgroups. Taken together with the impaired histological and immunohistochemical findings in the micro‐TESE(−) cases, these results suggest that these pathogens may play a role in testicular structural and functional changes. We performed microbiome analysis of testicular tissue using next‐generation sequencing (NGS) as the primary outcome. The association of microbiome analysis with histologic changes, ZO‐1 protein expression patterns, and micro‐TESE results were secondary outcomes. Conclusions: The testicular tissue microbiome may play a role inthe pathology of male infertility by contributing to structural and functional damageto the testis. It may predict negative micro‐TESE results and may also be a specific target for supportive care approaches, such as novel antimicrobial therapies, in NOA patients.

Objective: Poor responders pose a significant challenge in fertility treatment, leading to decreased oocyte quality and fertilization rates. Assisted oocyte activation (AOA) has emerged as a method to improve fertilization rates. This study aims to evaluate the effect of AOA on fertilization, pregnancy, and live birth rates in patients with diminished ovarian reserve. Materials and methods: This prospective non-randomized comparative study included a cohort of infertility patients with diminished ovarian reserve, divided into three groups: the control group receiving standard treatment (Group I/Conventional ICSI), the electrical activation group undergoing piezoelectric stimulation (Group II/ICSI + Piezoelectric), and the chemical activation group treated with calcium ionophore (Group III/ICSI + Calcium Ionophore). Each group consisted of 50 patients. The effects of electrical and chemical activation applied after ICSI on fertilization, pregnancy, and live birth rates were evaluated. Results: There were no statistically significant differences among the groups in hormone levels (FSH, LH, E2, PRL, and AMH), number of oocytes, or number of MII oocytes. Fertilization rates were significantly higher in the piezoelectric group compared to the control group ( p = 0.007). Although the fertilization rate in the calcium ionophore group was higher than that in the control group, this difference did not reach statistical significance ( p = 1.000). There was no statistically significant difference in fertilization rates between the piezoelectric and calcium ionophore groups ( p = 0.071). Regarding pregnancy rates, the highest implantation rate was observed in the calcium ionophore group (37%), but this difference does not reach statistical significance ( p = 0.224). No significant differences were observed among the groups in embryo quality, clinical pregnancy, ongoing pregnancy, and live birth rates. Conclusion: In patients classified as diminished ovarian reserve, artificial oocyte activation (AOA) was associated with higher fertilization rates for increasing fertilization rates, potentially resulting in a greater number of fertilized oocytes and transferable embryos. However, this apparent benefit does not translate into improved biochemical pregnancy outcomes. Moreover, no significant differences were observed between groups in terms of embryo quality, clinical pregnancy, ongoing pregnancy, and live birth rates. These findings suggest that while AOA may enhance early laboratory outcomes, its impact on clinically meaningful reproductive endpoints remains limited. Therefore, further adequately powered randomized studies are required to clarify the efficacy and safety of AOA.

To investigate microRNA-369-5p (miR-369-5p) and miR-671-3p expression in maternal peripheral blood leukocytes, plasma, and placental tissue in women with intrahepatic cholestasis of pregnancy (ICP) undergoing intracytoplasmic sperm injection (ICSI), compared with spontaneous conception. In this exploratory cohort study, microRNA-369-5p (miR-369-5p) and microRNA-671-3p (miR-671-3p) expression levels were analyzed using real-time quantitative PCR in maternal peripheral blood leukocytes, plasma, and placental tissue in four groups: (1) women with uncomplicated pregnancies conceived via intracytoplasmic sperm injection (non-ICP-ICSI, n = 7), (2) women with intrahepatic cholestasis of pregnancy conceived via intracytoplasmic sperm injection (ICP-ICSI, n = 7), (3) women with intrahepatic cholestasis of pregnancy who conceived spontaneously (ICP-SC, n = 7), and (4) women with uncomplicated pregnancies conceived spontaneously (non-ICP-SC, n = 7). Relative expression levels were calculated using the RQ method. Leukocyte miR-369-5p expression was significantly upregulated in ICP-ICSI and non-ICP-ICSI groups compared with ICP-SC and non-ICP-SC groups (p = 0.0001). Plasma miR-671-3p expression levels were significantly decreased in ICSI groups compared with spontaneous conception groups (p = 0.029). No significant differences were observed in placental expression levels of miR-369-5p and miR-671-3p among the four groups (p > 0.05). In subgroup analysis, leukocyte miR-369-5p and miR-671-3p expression levels were significantly higher in the ICP-ICSI group compared with the ICP-SC group (p < 0.05). Serum fasting bile acid levels showed a significant positive correlation with placental miR-369-5p expression (r = 0.467; p = 0.012) and a negative correlation with plasma miR-369-5p expression (r = −0.413; p = 0.029). Our findings show compartment-specific differences in miR-369-5p and miR-671-3p expression across leukocytes and plasma in this exploratory cohort. These preliminary observations do not establish causality and require validation in larger, independent cohorts.