Sperm cryopreservation is an essential technique for male fertility preservation, especially in men who are undergoing medical treatment. Conventional cryopreservation methods face limitations such as oxidative stress, DNA fragmentation, and cytotoxicity associated with traditional cryoprotectants like dimethyl sulfoxide (DMSO). Recent breakthroughs have focused on improving post-thaw sperm viability with novel cryoprotectants and innovative freezing strategies. Prospective approaches include the use of amino acid-based cryoprotectants, deep eutectic solvents, and antioxidants that have been described to prevent oxidative damage and maintain DNA integrity. Vitrification, a high-speed freezing technique that prevents ice crystal formation, has demonstrated superior outcomes compared to conventional slow freezing. Moreover, the Direct Dropping Method, a cryoprotectant-free approach, has been introduced as a contamination-minimizing technique that preserves sperm functionality. Multiomics tools are also utilized to determine biomarkers for protocol optimization. Despite these advancements, cryoprotectant toxicity is a central challenge, emphasizing the necessity for safer agents. Future research must focus on long-term sperm functionality and individualized cryopreservation strategies to maximize reproductive outcomes. The current review highlights the challenges associated with sperm cryopreservation, explores innovative strategies and novel cryoprotectants, underscores the significance of maintaining DNA integrity, and proposes future research directions to improve fertility preservation outcomes.

Objective: To translate, adapt, and validate the Indonesian version of the Prenatal Health Behavior Scale.

Methods: This cross-sectional, cross-cultural adaptation study was conducted between September 2024 and October 2024 in Ngrambe and Sine, subdistricts in Ngawi, East Java, Indonesia. We selected participants using purposive convenience sampling and matched them with inclusion and exclusion criteria. We collected sociodemographic, Prenatal Health Behavior Scale, and anthropometrics (height, weight, body mass index, and middle-upper arm circumference) data. We analyzed the content validity using the content validity index and Gwet's chance-corrected Agreement Coefficient 2, face validity by pilot-testing on several pregnant women, and construct validity using exploratory factor analysis. We measured reliability using McDonald's omega coefficient.

Results: We recruited 183 pregnant women in this study (median age 28 years). The item-content validity index (I-CVI) of all items was 1.00, with Gwet's chance-corrected Agreement Coefficient 2 was 0.945. The face validity resulted in a clear statement of all items. The exploratory factor analysis showed the two-factor model best suited to the questionnaire. Omega coefficients for the overall scale, health-impairing, and health-promoting domains were 0.696, 0.507, and 0.678, respectively.

Conclusions: The Indonesian version of the Prenatal Health Behavior Scale is a valid and reliable instrument to assess prenatal health behavior in Indonesian-speaking pregnant women. Future studies may implement this scale in community and clinical settings.

Objective: To investigate the levels of seminal plasma exosomes and the expression of the SRD5A2 gene in Iraqi men with different types of male infertility (asthenozoospermia, oligozoospermia, and azoospermia), and to explore the regulatory role of selected microRNAs (miRNAs) in the modulation of SRD5A2 gene expression.

Methods: A total of 90 male participants were categorized into four groups: asthenozoospermia (n=24), oligozoospermia (n=24), azoospermia (n=18), and normozoospermia (n=24). Seminal plasma exosome levels were quantified using ELISA. RNA, including miRNAs, was extracted, followed by cDNA synthesis. The expression of SRD5A2 mRNA and five selected miRNAs (miR-6090, hsa-miR-5189-5p, miR-23a-5p, miR-1914-3p, and miR-4540) was evaluated using qRT-PCR. Correlation analyses were conducted between exosome levels, miRNA expression, and SRD5A2 expression.

Results: Infertile men exhibited significantly lower seminal exosome levels than normozoospermic men (P<0.01). The expression of SRD5A2 mRNA was decreased in asthenozoospermic and oligozoospermic men but elevated in azoospermic men. Elevated levels of hsa-miR-5189-5p, miR-6090, and miR-23a-5p were observed in infertile groups. Correlation analysis revealed a significant negative relationship between these miRNAs and SRD5A2 expression in asthenozoospermic and oligozoospermic men, but a positive correlation in azoospermic men. Exosome levels were also significantly correlated with the expression of these miRNAs, suggesting their role as molecular carriers in regulating gene expression.

Conclusions: This study highlights the potential role of seminal exosomes and specific miRNAs in regulating SRD5A2 gene expression among infertile men. The altered levels of exosomes and dysregulated miRNA expression, particularly hsa-miR-5189-5p and miR-6090, may serve as novel biomarkers for the diagnosis and management of male infertility. Further research is warranted to validate these findings and explore their therapeutic implications.

Objective: To investigate whether differences exist in DNA fragmentation levels and protamine deficiency in the sperm of obese men compared to those of overweight and proven fertile, normal weight men and evaluate the alterations related to reproductive hormones, semen parameters and their association with body mass index (BMI).

Methods: Participants in this observational study were divided into three groups based on their BMI: proven fertile, normal weight men (n=200); overweight men (n=200); and obese men (n=200). After 3 days of abstinence, seminal fluid was collected from all participants and analyzed. Blood samples were also collected on the same day for hormonal analysis. Sperm DNA fragmentation and protamine deficiency were also assessed.

Results: A total of 600 men with a mean age of (34.3±7.9) years were included in the study. Obese males (BMI 30 kg/m² or higher) exhibited significant reductions in semen volume, sperm concentration, total sperm motility percentage, progressive motility, non-progressive motility, and levels of total testosterone, follicle-stimulating hormone (FSH) and luteinizing hormone (LH) compared to overweight (BMI 25 to 29.9 kg/m²) and normal weight males (BMI 18.5 to 24.9 kg/m²) (P<0.001). Conversely, obese males showed a significant increase in prolactin level, sperm DNA fragmentation, and protamine deficiency compared to overweight and normal weight males (P<0.001). Significant negative correlations were demonstrated between BMI and sperm concentration, motility, total testosterone, FSH and LH (P<0.001), whereas prolactin, sperm DNA fragmentation and protamine deficiency were positively correlated with BMI (P<0.001).

Conclusions: This study provides clear evidence that obesity contributes potentially to male infertility by inducing sperm DNA fragmentation and protamine deficiency, as well as negatively impacting reproductive hormones and semen quality.

Objective: To investigate the protective effect of alpha-lipoic acid on lead-induced Sertoli cell injury and its potential molecular mechanisms using primary Sertoli cells from rooster testes.

Methods: Primary Sertoli cells from 6-week-old rooster testes were divided into four goups. They were treated with serum-free DMEM-F12 medium (the control group), 40 μM lead acetate, 100 μM alpha-lipoic acid, or 40 μM lead acetate plus 100 μM alpha-lipoic acid for 24 h, respectively. Cell viability, reactive oxygen species levels (DCFH-DA), antioxidant enzyme activities (catalase, superoxide dismutase), blood-testis barrier-related proteins, nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway components, proliferation/apoptosis markers (Western blot), and related gene expression (qPCR) were analyzed.

Results: Alpha-lipoic acid treatment significantly reduced lead-induced reactive oxygen species (ROS) production by 58.3% compared to the lead acetate group (P<0.05), enhanced catalase activity by 35.8% (P<0.05), and elevated lead-reduced PCNA expression by 47.2% (P<0.05). Lead acetate 40 μM plus 100 μM alpha-lipoic acid significantly increased Nrf2 expression by 78.9% and HO1/NQO1 expression by 97.5% and 89.4%, respectively (P<0.05), significantly upregulated the expression of Bcl2 protein and significantly downregulated Bax protein expression (P<0.05).

Conclusions: Alpha-lipoic acid enhances the antioxidant properties of Sertoli cells by activating the Nrf2 pathway, restores lead-induced excessive autophagy, and inhibits apoptosis, thus alleviating the damage caused by lead in Sertoli cells.