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Background: Infertility is a major health issue, affecting 15% of reproductive‐age couples with male factors contributing to 50% of cases. Asthenozoospermia (AS), or low sperm motility, is a common cause of male infertility with complex aetiology, involving genetic and metabolic alterations, inflammation and oxidative stress. However, the molecular mechanisms behind low motility are unclear. In this study, we used a metabolomics approach to identify metabolic biomarkers and pathways involved in sperm motility. Methods: We compared the metabolome and lipidome of spermatozoa of men with normozoospermia (n = 44) and AS (n = 22) using untargeted LC–MS and the metabolome of seminal fluid using 1H‐NMR. Additionally, we evaluated the seminal fluid redox status to assess the oxidative stress in the ejaculate. Results: We identified 112 metabolites and 209 lipids in spermatozoa and 27 metabolites in the seminal fluid of normozoospermic and asthenozoospermic men. PCA analysis of the spermatozoa's metabolomics and lipidomics data showed a clear separation between groups. Spermatozoa of asthenozoospermic men presented lower levels of several amino acids, and increased levels of energetic substrates and lysophospholipids. However, the metabolome and redox status of the seminal fluid was not altered inAS. Conclusions: Our results indicate impaired metabolic pathways associated with redox homeostasis and amino acid, energy and lipid metabolism in AS. Taken together, these findings suggest that the metabolome and lipidome of human spermatozoa are key factors influencing their motility and that oxidative stress exposure during spermatogenesis or sperm maturation may be in the aetiology of decreased motility in AS. [ABSTRACT FROM AUTHOR]

Aim: To assess the efficacy of intratesticular injection of autologous platelet‐rich plasma (PRP) in men with non‐obstructive azoospermia (NOA) and a history of failed microdissection‐testicular sperm extraction (mTESE) procedures. Methods: A prospective case series of a cohort study was conducted involving couples diagnosed with NOA. Patients with at least one failed mTESE procedure were included. Intratesticular PRP injection was performed using a standardized protocol. Follow‐up assessments included sperm analysis, hormonal evaluation, and in vitro fertilization (IVF) outcomes. Results: Data from 177 men with NOA were analyzed, with 135 patients meeting eligibility criteria. PRP treatment resulted in positive sperm retrieval rates of 27.5% in patients with one prior failed mTESE procedure and 16.4% in patients with two or more failed attempts. IVF outcomes showed fertilization rates of 86.4% and 100.0% in respective groups, with pregnancy rates of 36.8% and 22.2% per embryo transfer. Histopathological examination post‐mTESE revealed varied patterns, including Sertoli cell‐only syndrome and maturation arrest. Conclusions: Intratesticular PRP injection shows promise as a potential therapeutic approach for NOA patients with prior failed mTESE procedures, demonstrating improved sperm retrieval rates and favorable IVF outcomes. Further randomized controlled trials are warranted to validate these findings and refine the technique's efficacy in male infertility management to answer the question of whether PRP could significantly improve the second attempt retrieval rate. [ABSTRACT FROM AUTHOR]

Purpose: To explore the application possibility of macrocephalic sperm from a patient with 100% macrocephalic sperm and AURKC gene variations. Methods: We diagnosed a case of macrozoospermia with 100% macrocephalic sperm and 39.5% multi-tailed spermatozoa by morphological analysis. Whole-exome sequencing (WES) was used for the patient and his wife. Sanger sequencing technique was used to verify the AURKC mutations in the patient's parents and his offspring. Sperm's ploidy was tested by flow cytometry. The couple asked for intra-couple ART therapy. Results: The patient presented novel compound heterozygous AURKC mutations (c.434C > T, c.497A > T) by WES. Sanger sequencing validation showed that variant of c.434C > T was observed in his father and c.497A > T was observed in his mother. Flow cytometry revealed that there existed a certain proportion of haploid sperm. Macrocephalic spermatozoa whose heads were smaller than the diameter of injection needle were selected for microinjection. A singleton pregnancy was achieved after embryo transfer. Prenatal diagnosis revealed that the fetus had normal chromosomal karyotype. Sanger sequencing technique showed that the fetus carried a c.434C > T mutation in one AURKC allele. A 3730 g healthy male fetus was delivered at term. Conclusion: Our study reported a successful live birth from a patient with definite AURKC gene variants and may provide insights for such patients to choose donor sperm or their own sperm. [ABSTRACT FROM AUTHOR]

Purpose: Teratozoospermia is the main pathogenic factor of male infertility. However, the genetic etiology of teratozoospermia is largely unknown. This study aims to clarify the relationship between novel variations in TENT5D and teratozoospermia in infertile patients. Materials and methods: Two infertile patients were enrolled. Routine semen analysis of patients and normal controls was conducted with the WHO guidelines. Whole‐exome sequencing (WES) was conducted to identify pathogenic variants in the two patients. Morphology and ultrastructure analysis of spermatozoa in the two patients was determined by Papanicolaou staining, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The functional effect of the identified variants was analyzed by immunofluorescence staining and western blotting. The expression of TENT5D in different germ cells was detected by immunofluorescence staining. Results: Two new hemizygous variations, c.101C > T (p.P34L) and c.125A > T (p.D42V), in TENT5D were detected in two patients with male infertility. Morphology analysis showed abnormalities in spermatozoa morphology in the two patients, including multiple heads, headless, multiple tails, coiled, and/or bent flagella. Ultrastructure analysis showed that most of the spermatozoa exhibited missing or irregularly arranged '9+2' structures. Further functional experiments confirmed the abrogated TENT5D protein expression in patients. In addition, both p.P34L and p.D42V substitutions resulted in a conformational change of the TENT5D protein. We precisely analyzed the subcellular localization of TENT5D in germ cells in humans and mice. And we found that TENT5D was predominantly detected in the head and flagellum of elongating spermatids and epididymal spermatozoa. Conclusions: Our results showed further evidence of a relationship between TENT5D mutation and human male infertility, providing new genetic insight for use in the diagnosis and treatment of male infertility. [ABSTRACT FROM AUTHOR]

Simple Summary: The process of acquiring sexed sperm involves the separation of sperm with X and Y chromosomes, allowing for the production of offspring of the desired sex. However, this process can negatively affect the quality of semen due to biochemical and physical stresses. ProAKAP4 is a major component of the sperm flagellum structure. In this study, sexed bull semen samples were evaluated using the proAKAP4 biomarker for the first time. Based on CASA and flow cytometry analysis of spermatozoa after thawing and incubation for 3 h, proAKAP4 is considered a practical biomarker for estimating the longevity of sexed sperm quality. ProAKAP4 is a sperm structural protein that regulates motility through the PKA-dependent cAMP signaling pathway, which is synthesized as an X chromosome-linked member of the gene family. This study aims to determine the optimal level of proAKAP4 for evaluating sexed semen through investigating its relationship with the longevity of sperm quality in sexed Holstein bull sperm. A total of 30 sexed sperm samples (bearing X chromosomes) from 30 distinct Holstein bulls (n = 30) were analyzed. The frozen bull sperm samples were assessed for their proAKAP4 levels, mitochondrial membrane potential, plasma membrane and acrosome integrity (PMAI), and spermatozoa movement parameters at hours 0 and 3 after thawing. The proAKAP4 levels in the sexed sperm samples ranged from 16.35 to 72.10 ng/10 M spz, with an average of 37.18 ± 15.1 ng/10 M spz. A strong positive correlation was observed between proAKAP4 levels and total motility, progressive motility, PMAI, high mitochondrial membrane potential, VAP, and VCL values after 3 h of incubation, when compared to post-thaw analyses. The results also reveal that spermatozoa with proAKAP4 levels of ≥40 ng/10 M spz exhibit higher quality. In conclusion, the level of proAKAP4 in sexed sperm aligns with previous studies and shows potential as a biomarker for assessing the longevity of sexed sperm quality. [ABSTRACT FROM AUTHOR]

Biostimulation aims to optimize reproductive parameters as part of animal management practices by modulating animal sensory systems. Chemical signals, mostly known as pheromones, have a great potential in this regard. This study was conducted to determine the influence of short-term male rabbit exposure to different biological secretions, potentially pheromone-mediated, on reproductive parameters of males. Four groups of 18 males each were exposed to A) doe urine, B) 2-phenoxyethanol, C) doe vaginal swab, and D) distilled water (control), three times over a 2.5h exposure window, just before semen collection. Semen volume, sperm concentration and motility, as well as subpopulation analysis of the spermatozoa were assessed for each condition. Additionally, testosterone levels in blood samples were monitored at five time points over the 2.5 h exposure window. We found a higher percentage of motile, progressive, fast progressive and mid-progressive spermatozoa in any of the three experimental groups compared to the control group. In contrast, the semen volume and the percentage of immotile and non-progressive spermatozoa was generally higher in the control group. We then identified a higher proportion of a subpopulation of fast and progressive spermatozoa in groups A, B, and C compared to group D. Our data indicates that sperm motility increases when animals are exposed to specific biological fluids potentially containing pheromones, and that an increase in sperm volume does not correlate with an increase in spermatozoa concentration, progressiveness, and speed. Finally, no differences in testosterone levels were found among comparisons, although males of groups A and C (exposed to natural female biological fluids) showed a tendency towards higher testosterone levels. In conclusion, our results indicate that rabbit sperm quality increases upon exposure to the biological secretions proposed, thereby supporting further investigation into their molecular identity. This exploration could eventually pave the way for implementing the use of pheromones in rabbit husbandry to enhance reproductive and productive parameters in farmed rabbits. • Buck semen quality increases upon exposure to specific female biological fluids. • Doe urine, vaginal fluid, and 2-phenoxyethanol are potential biostimulators. • Concentration and progressiveness of spermatozoa were enhanced in exposed rabbits. • Exposure to doe urine and vaginal fluid tended to increase testosterone levels. • Pheromone biostimulation is a natural method to improve animal production. [ABSTRACT FROM AUTHOR]

Background: Spermatogenic maturation arrest is thought to be caused by epigenetic defects, specifically in chromatin remodeling and histone modification. This study evaluated the status of chromatin remodeling chromodomain helicase DNA binding protein 5 (CHD5) and histone modifications histone 4 lys-12 acetylation (H4K12ac) and histone 3 lys-9 trimethylation (H3K9me3) in human testicular biopsies, based on maturation arrest type. Materials and Methods: The cross-sectional study utilized 18 Bouin-fixed paraffin-embedded (BFPE) specimens prepared from residual tissue from routine laboratory tests of infertile patients. The expression of CHD5, H4K12ac, and H3K9me3 was examined through immunohistochemistry (IHC). The intensity was measured using ImageJ with IHC Profiler and StarDist plugins. Statistical analysis was performed using Python with Scipy.Stats module. The data were tested with Shapiro-- Wilk for normality and Levene test for homogeneity. The differences in the intensity of spermatogenic cells were assessed using Kruskal-Wallis and Mann-Whitney tests. A difference was considered statistically significant if P<0.05. Results: We found three types of maturation arrest, including Sertoli cell only (n=5), spermatocyte arrest (n=4), and spermatid arrest (n=9). CHD5 was positive in spermatogonia and round spermatids but absent in spermatocytes. The mean grey value (MGV) of CHD5 in spermatogonia was generally weak in spermatocyte arrest (157.4 ± 16.6) and spermatid arrest (155.3 ± 16.8), and there was no significant difference between them [P=0.49, 95% confidence interval (CI): (-4.3, 6), effect size (r): 0.02]. Although there was a significant difference in the expression of H3K9me3 and H4K12ac (P<0.001), both histone modifications were found in all observed spermatogenic cells. Conclusion: The expressions of CHD5, H3K9me3, and H4K12ac in different spermatogenic cell types produce similar results, indicating that they cannot be used as markers to determine the type of spermatogenic maturation arrest in humans. The significant finding in this research is the expression of CHD5 in human spermatogonia cells, which requires further study for elaboration. [ABSTRACT FROM AUTHOR]

Introduction: Phosphorylation reactions highlight the essential role of protein kinases in sperm motility. TSSK6 protein is thought to play a crucial role in this process. Indeed, sperm obtained from mice with the Tssk6-/-genotype are unable to carry out fertilization and exhibit decreased motility. This leads us to investigate the causes of asthenozoospermia by searching for polymorphisms in the TSSK6 gene. Therefore, the objective of this study is to identify polymorphisms in the TSSK6 gene in men affected by asthenozoospermia. Material & Methods: The methodology involved direct sequencing of spermatozoa DNA. Thirty ejaculates were analyzed, including 20 from asthenozoospermic men and 10 from normozoospermic men. Spermograms were performed according to WHO procedures. DNA extraction was carried out using the phenol/chloroform method followed by conventional PCR. The amplicons were sequenced using the Sanger method, and the sequences were analyzed with BioEdit software. The data were analyzed using Fisher and Mann-Whitney tests. Results: The results revealed mutations in the TSSK6 gene in both normospermic and asthenozoospermic ejaculates. The synonymous mutations c.690T>C (p.Tyr230Tyr) and c.372C>A (p.Arg124Arg) were the most frequent, occurring at rates of 50% and 33.33%, respectively. Analysis of the mutations using PolyPhen-2 indicated that all mutations observed in normozoospermic samples are benign and would not affect sperm quality. However, only the mutations described in asthenozoospermic samples are predicted to be damaging to the protein. Conclusion: In conclusion, mutations in the TSSK6 gene were observed in infertile men. Deleterious mutations in the TSSK6 protein are associated with asthenozoospermia. [ABSTRACT FROM AUTHOR]

This study was designed to compare the addition of lecithin (LC) and lecithin nanoparticles (LC-NPs) to a semen extender (control group – egg yolk, EY) during a storing period in relation to its effectiveness on buffalo sperm cryopreservation. Ejaculates were collected from six mature, healthy Egyptian buffalo bulls using an artificial vagina, and diluted with nine extenders. LC-NPs and LC were tested at concentrations of 0.25, 0.5, 1.00, and 1.50 µg/ml compared to the egg yolk extender. The results indicated an overall enhancement in sperm quality: increased progressive motility and improved hypo-osmotic swelling test (HOST), reduced instances of dead sperm, abnormal morphology and chromatin damage in the LC-NPs group compared to both LC and the control groups (P < 0.05), apart from group 0.25 µg/ml. Furthermore, LC-NPs demonstrated superior viability at a concentration of 1 µg/ml. This concentration exhibited lower rates of early and late apoptotic events, as well as reduced necrosis compared to lower concentrations. The results showed significantly increased (P < 0.05) antioxidant indices, including superoxide dismutase (SOD), glutathione peroxidase, and total antioxidant capacity, with the inclusion LC-NPs, particularly notable at concentrations of 1.00 and 1.50 µg/ml, surpassing the effects observed in other groups. In contrast, the levels of malondialdehyde decreased significantly (P < 0.05) with LC-NPs, particularly at 1.00 µg/ml in comparison to the LC and control groups. Moreover, the fortification of cryopreserved spermatozoa with 1 or 1.5 µg/ml LC-NPs resulted in the preservation of acrosomal and plasma membrane integrity, as well as normal ultrastructure. So, the addition of LC-NPs, particularly of 1 or 1.5 µg/ml, significantly enhanced sperm quality by improving antioxidant indices, reducing apoptosis, and preserving ultrastructure integrity of buffalo sperm after the post-thawing process. [ABSTRACT FROM AUTHOR]

Purpose: This study aimed to identify the genetic causes of male infertility and primary ciliary dyskinesia (PCD)/PCD-like phenotypes in three unrelated Han Chinese families. Methods: We conducted whole-exome sequencing of three patients with male infertility and PCD/PCD-like phenotypes from three unrelated Chinese families. Ultrastructural and immunostaining analyses of patient spermatozoa and respiratory cilia and in vitro analyses were performed to analyze the effects of SPEF2 variants. Intracytoplasmic sperm injection (ICSI) was administered to three affected patients. Results: We identified four novel SPEF2 variants, including one novel homozygous splicing site variant [NC_000005.10(NM_024867.4): c.4447 + 1G > A] of the SPEF2 gene in family 1, novel compound heterozygous nonsense variants [NC_000005.10(NM_024867.4): c.1339C > T (p.R447*) and NC_000005.10(NM_024867.4): c.1645G > T (p.E549*)] in family 2, and one novel homozygous missense variant [NC_000005.10(NM_024867.4): c.2524G > A (p.D842N)] in family 3. All the patients presented with male infertility and PCD/likely PCD. All variants were present at very low levels in public databases, predicted to be deleterious in silico prediction tools, and were further confirmed deleterious by in vitro analyses. Ultrastructural analyses of the spermatozoa of the patients revealed the absence of the central pair complex in the sperm flagella. Immunostaining of the spermatozoa and respiratory cilia of the patients validated the pathogenicity of the SPEF2 variants. All patients carrying SPEF2 variants underwent one ICSI cycle and delivered healthy infants. Conclusion: Our study reported four novel pathogenic variants of SPEF2 in three male patients with infertility and PCD/PCD-like phenotypes, which not only extend the spectrum of SPEF2 mutations but also provide information for genetic counseling and treatment of such conditions. [ABSTRACT FROM AUTHOR]

Purpose: To identify the genetic causes of multiple morphological abnormalities in sperm flagella (MMAF) and male infertility in patients from two unrelated Han Chinese families. Methods: Whole-exome sequencing was conducted using blood samples from the two individuals with MMAF and male infertility. Hematoxylin and eosin staining and scanning electron microscopy were performed to evaluate sperm morphology. Ultrastructural and immunostaining analyses of the spermatozoa were performed. The HEK293T cells were used to confirm the pathogenicity of the variants. Results: We identified two novel homozygous missense ARMC2 variants: c.314C > T: p.P105L and c.2227A > G: p.N743D. Both variants are absent or rare in the human population genome data and are predicted to be deleterious. In vitro experiments indicated that both ARMC2 variants caused a slightly increased protein expression. ARMC2-mutant spermatozoa showed multiple morphological abnormalities (bent, short, coiled, absent, and irregular) in the flagella. In addition, the spermatozoa of the patients revealed a frequent absence of the central pair complex and disrupted axonemal ultrastructure. Conclusion: We identified two novel ARMC2 variants that caused male infertility and MMAF in Han Chinese patients. These findings expand the mutational spectrum of ARMC2 and provide insights into the complex causes and pathogenesis of MMAF. [ABSTRACT FROM AUTHOR]

In recent decades, a decline in male fertility due to deterioration in sperm quality has been noted around the world. This is probably explained by the tendency to increase the diseases of the male genital organs and, as a result, the increase in the percentage of male infertility. In addition, there is a deterioration of the quantitative and qualitative indicators of spermograms in practically healthy men. The average number of sperm in the ejaculate of a healthy man has halved over the past 50 years, and the average volume of ejaculate has decreased by one third. It is known that the multicomponent composition of the internal male genital organs is in constant restructuring due to age changes, functional activity and the influence of various factors. That is why it is important to take into account both physiological and age-related changes in a man's ability to conceive. Diagnosis of male infertility includes clinical research methods and methods of laboratory-instrumental examination. Among the latter, the most important for finding out the functional state of the gonads and the fertilizing ability of sperm is the study of ejaculate. The object of our research was spermograms obtained during the examination of patients at the "Alternative Clinic" reproductive medicine clinic. The purpose of the work was using biometric analysis to the different indicators of men spermograms of younger, middle and older age groups with normozoospermia (N), asthenozoospermia (AZS), teratozoospermia (T) and azoospermia (A). The task of the research was to analyze the main indicators of spermograms of men of different age groups in normal and pathological conditions, to conduct a one-factor and two-factor variance analysis of the influence of the studied diseases and age factor. After analyzing the main indicators of spermograms of men of younger, middle and older age groups with asthenozoospermia, azoospermia, and teratozoospermia, it was established that the indicators that undergo the greatest deviations in the studied diseases are the mobility of spermatozoa according to categories A and B, the morphology of spermatozoa, the Farris fertility index, and the activity of spermatozoa. After conducting a one-factor variance analysis, we established that the share of the influence of the studied diseases on the overall variability of such indicators of spermograms as the mobility of spermatozoa according to category A and B and the morphology of spermatozoa in the spermograms of men of the studied age groups is within 63–98% of the total contribution (younger age group), 60–96% (middle age group) and 75–96% (older age group). The share of influence of unaccounted factors is within 2–40% of the total contribution. The share of the influence of the studied diseases in the overall variability of such indicators as the Farris index and sperm activity in the spermograms of men of all studied groups is significantly reduced and is within 22–44% of the total contribution. Instead, the influence of unaccounted factors (56–78% of the total contribution) is growing significantly. This may indicate the presence of concomitant diseases and other pathologies in the men who took part in the research. After conducting a two-factor variance analysis, we established that the shares of the influence of the studied diseases on the overall variability of such indicators of spermograms as motility of spermatozoa according to category A and B, morphology of spermatozoa, Farris fertility index and activity of spermatozoa are decisive and are within the range of 90.7–99.9% of the total contribution, the share of the influence of the age factor on the variability of spermogram indicators of men of different age groups is insignificant. [ABSTRACT FROM AUTHOR]

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Background. Sperm motility is a fundamental factor for sperm penetration into the oocyte and fertilization. According to the World Health Organization (WHO) 2021, less than 42 percent of motility is called asthenozoospermia, which is one of the most common causes of male infertility. This retrospective cohort study aims to investigate sperm DNA damage with TUNEL and SCSA methods and its relationship with sperm parameters, age, and body mass index (BMI) in severe asthenospermia (<2% sperm motility) and normozoospermia. Methods. The study parameters between 111 subjects with severe asthenozoospermia and 113 subjects with normozoospermia were investigated. The 2010 World Health Organization guidelines were used to evaluate sperm parameters, and TUNEL and SCSA methods were used to assess sperm DNA damage. The statistical analysis was done using the t-test of two independent samples. Pearson's correlation coefficient was used to investigate the relationship between sperm DNA damage and sperm parameters, age, and BMI P<0.05 was considered significant. Results. The mean seminal volume, sperm concentration and count, and the percentage of sperm total motility and progressive motility were significantly lower in severe asthenospermic subjects compared to normozoospermic subjects (P<0.001). In addition, the mean percentage of sperm DNA damage in severe asthenozoospermic individuals was significantly higher than in normozoospermic individuals (P<0.001). There was also a positive and significant relationship between sperm DNA damage and the age of severely asthenozoospermic men. Conclusion. Sperm motility, sperm DNA damage, and the father's age are essential in the successful conception, development, and health of the embryo. Therefore, the evaluation of sperm DNA damage is recommended in examining male fertility and selecting the appropriate treatment approach for men with severe asthenozoospermia. Practical Implications. Considering that the sperm genome constitutes 50 percent of the genetic material of the next generation, evaluating sperm DNA health to determine the appropriate treatment approach in infertile men can be influential in the success of assisted reproductive techniques and in maintaining the next generation’s health. [ABSTRACT FROM AUTHOR]

The demand for regenerative medicine products is growing rapidly in clinical practice. Unfortunately, their use has certain limitations. One of these, which significantly constrains the widespread distribution and commercialization of such materials, is their short life span. For products containing suspensions of cells, this issue can be solved by using cryopreservation. However, this approach is rarely used for multicomponent tissue-engineered products due to the complexity of selecting appropriate cryopreservation protocols and the lack of established criteria for assessing the quality of such products once defrosted. Our research is aimed at developing a cryopreservation protocol for an original hydrogel scaffold with encapsulated MSCs and developing a set of criteria for assessing the quality of their functional activity in vitro. The scaffolds were frozen using two alternative types of cryocontainers and stored at either −40 °C or −80 °C. After cryopreservation, the external state of the scaffolds was evaluated in addition to recording the cell viability, visible changes during subsequent cultivation, and any alterations in proliferative and secretory activity. These observations were compared to those of scaffolds cultivated without cryopreservation. It was shown that cryopreservation at −80 °C in an appropriate type of cryocontainer was optimal for the hydrogels/adipose-derived stem cells (ASCs) tested if it provided a smooth temperature decrease during freezing over a period of at least three hours until the target values of the cryopreservation temperature regimen were reached. It was shown that evaluating a set of indicators, including the viability, the morphology, and the proliferative and secretory activity of the cells, enables the characterization of the quality of a tissue-engineered construct after its withdrawal from cryopreservation, as well as indicating the effectiveness of the cryopreservation protocol. [ABSTRACT FROM AUTHOR]

Background: Sperm DNA fragmentation (SDF) can affect fertilization rate and embryo development, making it a useful measure for assessing male fertility. Available evidence supports the association between high sperm DNA fragmentation and poor outcomes, with regard to natural conception. Several treatment options are being adopted with varying degrees of success. Some of the commonly used treatment options are the intake of oral antioxidants, varicocele repair, and techniques like micromanipulation-based sperm selection and use of testicular sperm for intracytoplasmic sperm injection. Case Presentation: Studies have shown that around 29% of couples depend on complementary and alternative medicine (CAM) modality for the treatment of infertility. However, there is a lack of substantial evidence regarding its efficacy in treating various aspects of infertility in couples. The current case report is about a 44 year-old male patient with infertility, who has a known diagnosis of sex chromosome abnormalities. Meanwhile, the SDF study reports indicated the presence of chromosomal abnormalities. This patient was treated exclusively with Ayurveda therapy aimed towards qualitative improvement in reproductive tissues (Shukra Dhatu as per Ayurveda). Patient was assessed periodically for changes in chromosomal abnormality. After four months of treatment, the evaluations demonstrated the presence of completely normal chromosomes. Conclusion: This case study indicates the potential of Ayurveda therapy in treating cases of male infertility caused by DNA fragmentation. Furthermore, observations and systematically designed clinical trials are warranted to establish a stronger level of evidence before making further clinical recommendations. [ABSTRACT FROM AUTHOR]

Purpose: The advent of proteomics provides new opportunities to investigate the molecular mechanisms underlying male infertility. The selection of relevant targets based on a single analysis is not always feasible, due to the growing number of proteomic studies with conflicting results. Thus, this study aimed to systematically review investigations comparing the sperm proteome of normozoospermic and infertile men to define a panel of proteins with the potential to be used to evaluate sperm quality. Materials and Methods: A literature search was conducted on PubMed, Web of Science, and Scopus databases following the PRISMA guidelines. To identify proteins systematically reported, first the studies were divided by condition into four groups (asthenozoospermia, low motility, unexplained infertility, and infertility related to risk factors) and then, all studies were analysed simultaneously (poor sperm quality). To gain molecular insights regarding identified proteins, additional searches were performed within the Human Protein Atlas, Mouse Genome Informatics, UniProt, and PubMed databases. Results: Thirty-two studies were included and divided into 4 sub-analysis groups. A total of 2752 proteins were collected, of which 38, 1, 3 and 2 were indicated as potential markers for asthenozoospermia, low motility, unexplained infertility and infertility related to risk factors, respectively, and 58 for poor sperm quality. Among the identified proteins, ACR, ACRBP, ACRV1, ACTL9, AKAP4, ATG3, CCT2, CFAP276, CFAP52, FAM209A, GGH, HPRT1, LYZL4, PRDX6, PRSS37, REEP6, ROPN1B, SPACA3, SOD1, SPEM1, SPESP1, SPINK2, TEKT5, and ZPBP were highlighted due to their roles in male reproductive tissues, association with infertility phenotypes or participation in specific biological functions in spermatozoa. Conclusions: Sperm proteomics allows the identification of protein markers with the potential to overcome limitations in male infertility diagnosis and to understand changes in sperm function at the molecular level. This study provides a reliable list of systematically reported proteins that could be potential targets for further basic and clinical studies. [ABSTRACT FROM AUTHOR]

Conventional sperm selection based on motility and morphology fails to provide detailed information on sperm functional and molecular status. Magnetic-activated cell sorting (MACS) protocol aims to optimize this process by selecting apoptotic sperm cells. Phospholipase C zeta-1 (PLCz1) is a physiological stimulus for oocyte activation and early embryonic development. The purpose of this study was to examine seminal parameters, DNA fragmentation index (DFI), and PLCz1 expression levels in MACS-DGC sorted specimens (DFI > 30%) and assess early development in resulting embryos. Semen specimens from 60 patients diagnosed with male factor infertility were collected and processed by either density gradient centrifugation (DGC) or MACS-DGC protocols. Pre and post-preparation analysis was performed. PLCz1 expression was assessed using the RT-PCR method. Retrieved eggs from their partners were divided into two groups in which they were injected with different sorted sperm. The fertilization rate and embryonic development were evaluated. While sperm's progressive motility and morphology significantly improved, there was a substantial decline in DFI following MACS-DGC. Fertilization rates were almost the same between the groups, and the latter resulted in remarkably more top-quality embryos and more blastocysts. PLCz1 expression was considerably higher in the MACS-DGC group. By eliminating apoptotic cells, the MACS-DGC technique could sort highly PLCz1-expressed sperm, optimize sperm selection in individuals with elevated DFI, development of resulting embryos. [ABSTRACT FROM AUTHOR]

Electronic cigarettes that are currently popular are used as nicotine replacement therapy. There are several substances contained in electronic cigarette liquid that have carcinogenic effects. These substances can also increase free radicals which in turn can cause oxidative stress. Nicotine, propylene glycol, and glycerol can cause disturbances in male reproductive function. Infertility can occur due to lifestyle factors, one of which is smoking. Nicotine found in cigarettes can reduce sperm quality, including concentration, mobility, viability, and sperm morphology. The purpose of this study was to determine the effect of electronic cigarettes on sperm quality. The research method uses a literature review system. The twelve studies that are in accordance with the topic of discussion are experimental studies on mice and rats, two studies on human, and one study on both human and rats. Five journals showed a significant decrease in sperm count and testosterone serum level, three other journals showed a significant decrease in sperm morphology and viability, six journals showed a decrease in sperm motility, seven journals showed a significant impairment of testis histological structure, four journals showed a decrease of steroidogenesis enzymes, and one journal showed a significant increase of DNA fragmentation due to the use of electronic cigarettes. Electronic cigarettes had an effect on sperm quality. [ABSTRACT FROM AUTHOR]

Background: As the impacts of diabetes-induced reproductive damage are now evident in young people, we are now in urgent need to devise new ways to protect and enhance the reproductive health of diabetic people. The present study aimed to evaluate the protective effects of enalapril (an ACE inhibitor) and paricalcitol (a vitamin D analog), individually or in combination, on streptozotocin (STZ)-diabetes-induced testicular dysfunction in rats and to identify the possible mechanisms for this protection. Material and methods: This study was carried out on 50 male Sprague-Dawley rats; 10 normal rats were allocated as a non-diabetic control group. A total of 40 rats developed diabetes after receiving a single dose of STZ; then, the diabetic rats were divided into four groups of equivalent numbers assigned as diabetic control, enalapril-treated, paricalcitol-treated, and combined enalapril-and-paricalcitol-treated groups. The effects of mono and combined therapy with paricalcitol and enalapril on testicular functions, sperm activity, glycemic state oxidative stress, and inflammatory parameters, as well as histopathological examinations, were assessed in comparison with the normal and diabetic control rats. Results: As a result of diabetes induction, epididymal sperm count, sperm motility, serum levels of testosterone, follicle-stimulating hormone (FSH) as well as luteinizing hormone (LH), and the antioxidant enzyme activities, were significantly decreased, while abnormal sperm (%), insulin resistance, nitric oxide (NO), malondialdehyde (MDA), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were significantly increased, along with severe distortion of the testicular structure. Interestingly, treatment with paricalcitol and enalapril, either alone or in combination, significantly improved the sperm parameters, increased antioxidant enzyme activities in addition to serum levels of testosterone, FSH, and LH, reduced insulin resistance, IL-6, and TNF-α levels, and finally ameliorated the diabetes-induced testicular oxidative stress and histopathological damage, with somewhat superior effect for paricalcitol monotherapy and combined therapy with both drugs compared to monotherapy with enalapril alone. Conclusions: Monotherapy with paricalcitol and its combination therapy with enalapril has a somewhat superior effect in improving diabetes-induced testicular dysfunction (most probably as a result of their hypoglycemic, antioxidant, anti-inflammatory, and anti-apoptotic properties) compared with monotherapy with enalapril alone in male rats, recommending a synergistic impact of both drugs. [ABSTRACT FROM AUTHOR]

Detection of sperm in the peritoneal fluid of animals is unusual and has not been reported in the literature. In this report, we describe the presence of sperm cells in the peritoneal fluid of a twoyear-old ram. The ram was presented with dyspnea, reduced rumen contractions, a mild degree of dehydration, cyanotic mucosa, difficulty in standing, and anuria. Ancillary diagnostics, including ultrasonography, radiography, complete blood cell count, and abdominocentesis were performed. In the peripheral blood sample, no blood parasites were observed, although thrombocytosis with toxic neutrophilia, and atypical lymphocytes were seen in the blood smear. In radiographs, urolithiasis was confirmed just on the sigmoid flexure position. Ultrasound examination revealed a distended bladder and large amounts of free fluid within the peritoneal cavity. The electrocardiogram analysis showed the absence of P waves, bradycardia, wide QRS complexes, ST-segment elevation, and tented T waves. In the peritoneal fluid smears, a large number of spermatozoa, and increased inflammatory cells were observed. It was concluded that the trauma or rupture in colliculus seminalis, ductus deferens, or the urethra with urinary stones resulted in leakage of spermatozoa to the peritoneal cavity. To our knowledge, this is the first report of the presence of spermatozoa in the peritoneal fluid of large animals. [ABSTRACT FROM AUTHOR]

The results of artificial insemination (AI) are adversely affected by changes in sperm motility and function throughout the cryopreservation procedure. The proteome alterations of frozen–thawed spermatozoa with various levels of freezability in dairy goats, however, remain largely unknown. To discover differentially expressed proteins (DEPs) and their roles in dairy goat sperm with high or low freezability (HF or LF), we conducted 4D-DIA quantitative proteomics analysis, the results of which are presented in this work. Additionally, we explored the underlying processes that may lead to the variations in sperm freezing resistance. A total of 263 DEPs (Fold Change > 2.0, p-value < 0.05) were identified between the HF group and LF group in frozen–thawed dairy goat spermatozoa. In our Gene Ontology (GO) enrichment analysis, the DEPs were mostly associated with the regulation of biological processes, metabolic processes, and responses to stress and cellular component biogenesis. Our Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis also revealed that the DEPs were predominantly engaged in oxidative phosphorylation, N-Glycan biosythesis, and cysteine and methionien metabolism. A protein–protein interaction (PPI) network analysis revealed 14 potential proteins (NUDFB8, SDHC, PDIA4, HSPB1, etc.) that might influence the freezability of dairy goat sperm. These findings shed light on the processes underlying alterations in the proteome and sperm freezability, aiding further research on sperm cryopreservation. [ABSTRACT FROM AUTHOR]

Background/Aim: Sperm quality has experienced a decline in recent years, with this issue being particularly pronounced in industrialized nations, suggesting a potential link to occupational exposures. Evaluating sperm DNA fragmentation can yield valuable insights into male fertility, although its association with occupational exposures remains less well-established. Our study aimed to investigate the relationship between self-reported occupational exposures and sperm DNA fragmentation in infertile men. Methods: This retrospective cohort study involved 391 infertile men who sought fertility treatment at a university clinic between 2017 and 2020. A brief questionnaire was administered to collect data on patients’ demographic characteristics, medical history, occupation, and exposure types. In this comparative study, patients were categorized into two groups based on their occupational exposures (the unexposed and exposed groups). The exposed group was further sub-grouped according to their specific exposure types, which included cement, solvents, metals, pesticides, mechanical vibration, and heat. The primary outcome in this study was assessed using the terminal deoxynucleotidyl transferase-mediated nick end-labeling test (TUNEL), with results expressed as the sperm DNA fragmentation index (DFI). Results: Patients in the exposed group exhibited a significantly higher sperm DFI compared to those in the unexposed group (14 [17] vs. 8 [9], P<0.001). After accounting for potential confounding factors, our results demonstrated that several occupational exposure factors significantly increased the risk of elevated sperm DFI (>15%) levels, including solvents (odds ratio (OR) 8.2, 95% confidence interval (CI) 3.6–18.5, P<0.001), metals (OR: 2.2, 95% CI: 1.0–4.7, P=0.048), pesticides (OR: 14.6, 95% CI: 1.6–130.7, P=0.016), mechanical vibration (OR: 2.6, 95% CI: 1.5–4.6, P<0.001), and heat (OR: 6.4, 95% CI: 1.7–23.5, P=0.005). Conclusion: The findings of our study corroborate earlier research suggesting that occupational exposures may have adverse effects on sperm DNA fragmentation in men. The identification and management of such exposures as part of routine clinical practice could offer a complementary approach to enhancing infertility treatment outcomes. [ABSTRACT FROM AUTHOR]

Background: Previous studies examined the effects of obesity on sperm parameters and reported inconsistent results. Thus, the present study aimed to evaluate the association between obesity and the quality of sperm parameters in infertile men. Material and methods: The present cross-sectional study evaluated 218 infertile men aged 20–50. To this end, the 168-item food frequency questionnaire (FFQ) was utilized to evaluate dietary intake. The anthropometric and biochemical variables were examined using standard methods. Further, the association between obesity and the quality of sperm parameters was evaluated using the controlled linear regression for potential confounders. Results: The normal sperm morphology had a significant inverse association with BMI [adjusted β − 0.074, CI (− 0.141 to − 0.008), P = 0.029] and WC [adjusted β − 0.026, CI (− 0.051 to − 0.001), P = 0.038]. Additionally, visceral fat had a marginal inverse association with normal sperm morphology [adjusted β − 0.065, CI (− 0.138 to 0.008), P = 0.079] and non-progressive sperm motility [adjusted β − 0.241, CI (− 0.495 to 0.014), P = 0.063]. Conclusion: Even though the present results indicated that obesity, abdominal obesity, and visceral fat had inverse associations with normal sperm morphology, more mechanism-based studies should be conducted to confirm these findings. [ABSTRACT FROM AUTHOR]

Testicular dysfunction, a severe secondary complication of diabetes, negatively affects reproductive health. Ghrelin is a peptide that plays a role in many metabolic events, such as stimulating growth hormones and regulating appetite, blood pressure, and reproduction. Metformin has antiinflammatory and antioxidant properties and is widely used to treat type 2 diabetes. Therefore, in this study, to investigate the effects of ghrelin and metformin on testicular function in experimental diabetic rats given streptozotocin, placed on a high-fat diet, and that had their sperm parameters evaluated, testosterone hormone, caspase 3 and iNOS expression, and histopathological examinations were performed. In total, 38 Wistar albino male rats were randomized into 4 groups (a control group, diabetes group, diabetes + ghrelin group, and diabetes + metformin group) with different numbers. It was determined that spermatozoa concentration, motility, and testosterone levels were lower (p < 0.001), caspase 3 and iNOS expression levels (p < 0.05) were higher, and abnormal sperm rates were higher (p < 0.001) in the diabetic group compared to the other groups. Metformin and ghrelin administered to the diabetes group increased spermatozoa concentration, motility, and testosterone levels (p < 0.001) and decreased caspase 3 and iNOS expression levels, as well as abnormal sperm rates (p < 0.05). Based on these findings, we propose that ghrelin and metformin can be used to protect reproductive health in testicular dysfunction caused by diabetes. [ABSTRACT FROM AUTHOR]

Background: Semen hyperviscosity is a threatening cause of abnormal spermatozoa and infertility in men. We aimed to evaluate oxidative stress, antioxidants depletion and sperm apoptosis as main reasons for poor quality of spermatozoa in men with hyperviscous semen. Materials and Methods: In this cross-sectional study, ejaculate specimens were collected from fertile (n=102) and infertile men with hyperviscous semen (n=123) and without semen hyperviscosity (n=143). Total antioxidant capacity (TAC), glutathione (GSH), malondialdehyde (MDA), protein carbonyl (PC), 8-hydroxydeoxyguanosine (8-OHdG), and were measured in semen samples to estimate oxidative stress status. Gene expression pattern of BAX, CASPASE-9, CASPASE-3, and BCL2 was assessed to estimate sperm apoptosis. Results: The average of sperm count, normal morphology, normal motility, and sperm vitality in men with hyperviscous semen was significantly lower than infertile subjects without hyperviscous semen (P<0.01). Men with hyperviscous semen exhibited higher levels of PC (8.34 ± 1.03 nmol/mg vs. 6.01 ± 0.93 nmol/mg, P=0.008), MDA (1.14 ± 0.27 nmol/ ml vs. 0.89 ± 0.22 nmol/ml, P=0.031), 8-OHdG (259.71 ± 24.59 ng/ml vs. 197.13 ± 18.47 ng/ml, P=0.009), but lower TAC contents (1250.44 ± 66.23 μM/L vs. 1784.31 ± 89.87 μM/L, P=0.018) and GSH (3.82 ± 1.05 μM vs. 5.89 ± 0.87 μM, P=0.021) than men with non-viscous semen. The expression of BAX, CASPASE-3 and CASPASE-9 genes in men with hyperviscous semen was significantly increased by 1.39-fold (P=0.041), 1.47-fold (P=0.046), 1.29-fold (P=0.048), respectively, as compared with the infertile subjects without hyperviscous semen. However, BCL2 expression in infertile men without hyperviscous semen was higher compared to those with hyperviscous semen (1.36-fold, P=0.044). Conclusion: Hyperviscous semen is markedly associated with depletion of seminal plasma antioxidants, sperm membrane lipid peroxidation, DNA and protein oxidation, and sperm apoptosis. Antioxidant therapy might be considered as a valuable strategy to protect sperm cells against oxidative damage in cases with seminal fluid hyperviscosity. [ABSTRACT FROM AUTHOR]

Background: The objective of the current study was comparing the impact of two staining techniques on semen morphological parameters and their influence on patient diagnosis. The ideal staining method should preserve cell integrity while providing detailed information. Methods: Semen samples from fifty men were stained using Diff-Quick or Spermac methods. Morphological parameters were classified based on the Tygerberg criteria, and final diagnosis was according to WHO manual guidelines. Statistical analysis was performed through conducting paired t-tests or Wilcoxon rank-sum tests, with GLIMMIX and Fisher's exact test for determining the significance (p≤0.05). Results: Both staining methods highlighted head and tail regions, with Spermac offering better visualization of the midpiece. Spermac demonstrated fewer normal spermatozoa (2.8±0.3%) compared to Diff-Quick (3.98±0.4%; p=0.0385). Midpiece abnormalities were more evident with Spermac (55.7±2.1%) than Diff-Quick (24.8±2.0%; p<0.0001). No significant difference was found in head and tail abnormalities (p>0.05). Conclusion: Diff-Quick staining resulted in a higher proportion of normal spermatozoa, primarily due to its midpiece evaluation. The choice of staining method significantly impacts the diagnosis of infertile males. These findings have important implications for clinical practice and future research, suggesting the need for further investigations to assess different staining methods and determine optimal diagnostic thresholds. [ABSTRACT FROM AUTHOR]

Background: Sox2 (SRY box2) is an essential transcription factor that plays a vital role in spermatogenesis and regulates the genes in this process. Sox2 is important for pluripotency, self-renewal, and even spermatogonial stem cell differentiation. This gene is found in pluripotent and specialized cells, and it is involved in their biological activities. Methods: Protein-protein interaction (PPI) network analysis was performed during spermatogenesis using NCBI, STRING, and Cytoscape databases. Then, after isolating spermatogonial stem cells from 6 C57BL/6 mice, mouse embryonic stem cells and ES-like cells were prepared. In the following, Sox2 expression was examined in differentiated and undifferentiated spermatogonia by immunohistochemistry (IMH), immunocytochemistry (ICC), and Fluidigm PCR (polymerase chain reaction). Finally, the results were compared using the Kruskal-Wallis and Dunn tests at the significance level of p<0.05. Results: The results of this experiment showed that contrary to expectations, Sox2 has cytoplasmic expression in undifferentiated cells and nuclear expression in differentiated cells in in vitro conditions. In addition, the expression of Sox2 increased during differentiation. Fluidigm PCR showed a significantly higher expression of Sox2 (p<0.05) in differentiated compared to undifferentiated spermatogonia. Sox2 has an interaction with other genes during spermatogenesis such as Oct4, Nanog, Klf4, Stra8, Smad1, Tcf3, and Osm. Conclusion: Sox2, which is known as a pluripotency marker, has a vital role in spermatogenesis and could be a differential marker. Sox2 has strong connections with other genes such as Oct4, Nanog, Klf4, Tcf3, Osm, Stra8, Lim2, Smad1, Gdnf, and Kit. [ABSTRACT FROM AUTHOR]

The effects of diet and nutritional habits on reproductive cells can be categorized in a variety of ways. In this review, the literature is divided, based on the dietary consumption effects on oocytes and sperm. Topics on dietary patterns and the intrauterine effect of maternal nutrition are covered. In general fruits, vegetables, whole greens, fish, legumes, and also dietary sources containing unsaturated fats can improve reproductive germ cell quality. In epidemiological studies, the food intake frequency questionnaire is one of the most common methods to assess diet. Due to methodological heterogeneity in dietary assessment and inadequacy in the measurement of dietary intake in the questionnaires used, several unreliable results may be reported. Thus, the quality of evidence needs to be improved, since nutritional diets may not be so simply objective and they are inadequate to explain obvious underlining mechanisms. In addition, various compounds that may be ingested can affect molecular mechanisms, influenced by other external factors (drugs, pesticides, smoking, alcohol) and changes in human nutritional parameters. Artificial Intelligence has recently gained widespread interest and may have a role in accurate analysis of dietary patterns for optimal nutritional benefit. Therefore, future prospective randomized studies and objective measurements, consisting of molecular level analysis of the impact on cells and clear-cut methods are needed for accurate assessment of the effect of dietary habits on reproductive treatment. [ABSTRACT FROM AUTHOR]

Copyright of African Journal of Reproductive Health is the property of Women's Health & Action Research Centre and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

For over 30 years, defects of the functional integrity of the sperm membrane, as evidenced by a low hypo-osmotic swelling test when evaluating the semen analysis, are not only associated with male infertility (even with sperm that otherwise seem normal), but unless corrected, successful intrauterine pregnancies will rarely ensue. This defect, interestingly, does not impair fertilization of the oocyte, but instead, prevents a normal-appearing embryo from successfully implanting. The frequency in infertile couples increases with advancing age of the male, ranging from 5% in younger males to 25% in men in their late forties or early fifties. It seems to be related to a toxic protein added to the sperm as they traverse the ejaculatory ducts. The defect is very correctable, either by treating the sperm with the protein digestive enzyme chymotrypsin prior to intrauterine insemination and avoidance of unprotected sex prior to ovulation, or in vitro fertilization with intracytoplasmic sperm injection. Unfortunately, this very inexpensive, easy-to-perform test is rarely performed by the large majority of physicians treating infertility. The purpose of this manuscript is to hopefully rekindle interest within the infertility community to add this test to the standard semen analysis. [ABSTRACT FROM AUTHOR]

Spermatozoa are unique cells that carry a library of proteins that regulate the functions of molecules to achieve functional capabilities. Currently, large amounts of protein have been identified in spermatozoa from different species using proteomic approaches. However, the proteome characteristics and regulatory mechanisms of spermatozoa in bucks versus rams have not been fully unraveled. In this study, we performed a tandem mass tag (TMT)-labeled quantitative proteomic analysis to investigate the protein profiles in the spermatozoa of buck (Capra hircus) and ram (Ovis aries), two important economic livestock species with different fertility potentials. Overall, 2644 proteins were identified and quantified via this approach. Thus, 279 differentially abundant proteins (DAPs) were filtered with a p-value < 0.05, and a quantitative ratio of >2.0 or <0.5 (fold change, FC) in bucks versus rams, wherein 153 were upregulated and 126 were downregulated. Bioinformatics analysis revealed that these DAPs were mainly localized in the mitochondria, extracellular and in the nucleus, and were involved in sperm motility, membrane components, oxidoreductase activity, endopeptidase complex and proteasome-mediated ubiquitin-dependent protein catabolism. Specifically, partial DAPs, such as heat shock protein 90 α family class a member 1 (HSP90AA1), adenosine triphosphate citrate lyase (ACLY), proteasome 26S subunit and non-ATPase 4 (PSMD4), act as "cross-talk" nodes in protein–protein networks as key intermediates or enzymes, which are mainly involved in responses to stimuli, catalytic activity and molecular function regulator pathways that are strictly related to spermatozoa function. The results of our study offer valuable insights into the molecular mechanisms of ram spermatozoa function, and also promote an efficient spermatozoa utilization link to fertility or specific biotechnologies for bucks and rams. [ABSTRACT FROM AUTHOR]

Background: Sperm DNA fragmentation can affect reproductive outcomes in assisted reproductive techniques (ART), and it is a concern in density gradient centrifugation (DGC). By contrast, microfluidic approaches allow the selection of highly motile sperm with low DNA fragmentation index (DFI). The purpose of current study, was to compare the efficacy of a microfluidic device designed in-house in comparison with DGC. Methods: Nineteen healthy men with normal semen profiles were included in the study. Semen samples were individually aliquoted for three sperm preparation analyses (crude and processed with to either DGC or the microfluidic method). Sperm parameters of the samples were evaluated along with DNA fragmentation using the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) method. Results: Sperm processed using the microfluidic method showed a significantly lower DFI than those obtained using DGC and in crude semen, with DFI of 1.1%, 3.5%, and 4.9%, respectively. Although the microfluidic method yielded significantly lower sperm concentrations than DGC, no significant differences were observed in total motility, progressive motility, curvilinear velocity, straight-line velocity, or normal morphology. Conclusion: Using the in-house microfluidic device, sperm with lower DFI was effectively isolated when compared with DGC. The motility and normal morphology rates were comparable among the samples. [ABSTRACT FROM AUTHOR]

Copyright of Revista de la Facultad de Medicina Humana is the property of Instituto de Investigaciones en Ciencias Biomedicas de la Universidad Ricardo Palma and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: There are considerable opium addicts, particularly among workers in Iran's industrial sectors. This study aimed to investigate the effects of simultaneous exposure to excessive noise and opium addiction on adult rat sperm parameters. Materials and Methods: In this experimental study 42 adult Wistar male rats in 6 groups were studied as follows: group 1 control, group 2 exposed to noise (100 dB) 8 h/day, group 3 received daily 50 mg/case opium solution, group 4 received daily 100 mg/case opium solution, group 5 received daily 50 mg/case opium solution and exposed to noise 8h/day, and group 6 received daily 100 mg/case opium solution and exposed to noise 8h/day. After 50 days, cauda epididymis was removed for sperm parameters examination (WHO guidelines). Results: In the groups receiving opium (50 and 100 mg/case), sperm count, sperm viability, and normal sperm morphology (%) significantly decreased compared to the control group. Exposure to noise and consumption of opium solution simultaneously significantly reduced the count, viability, and percentage of sperm with normal morphology in both selected doses, as well as a significant difference between groups 5 and 6 in count, viability, and normal sperm morphology(%) parameters(P<0.05). Conclusions: According to the results, noise exposure and taking opium would actively reduce the count, viability, and normal sperm morphology. It is therefore suggested that the mechanism of such effects should be investigated in animal and human studies. [ABSTRACT FROM AUTHOR]

Oxidative stress due to low temperatures during in vitro preservation reduces boar spermatozoa quality. It has been proven that Schisandrin B (Sch-B) can act against oxidative stress in cells. Therefore, the purpose of this study was to investigate whether the treatment with Sch-B could improve the quality of boar sperm during storage at 17 °C. Semen samples were randomly divided into four groups and added to the Beltsville Thawing Solution containing different concentrations of Sch-B (0, 0.1, 0.5, and 1 mg/L) after collection. Each group was then preserved at 17 °C and the sperm motility, membrane integrity, and acrosome integrity were detected to determine the maximum available concentration of Sch-B for sperm. The optimal concentration was set at 0.1 mg/L and was used in subsequent experiments. Sperms treated with 0 and 0.1 mg/L Sch-B were evaluated for lipid peroxidation (MDA) and fertilization ability through in vitro fertilization. Finally, the quality of blastocysts which were formed by 0 and 0.1 mg/L Sch-B-treated sperm was determined. The results showed that compared with the control, the addition of 0.1 mg/L Sch-B improved boar sperm motility, and the addition of 0.1 and 0.5 mg/L Sch-B improved sperm membrane integrity and acrosome integrity. Treatment with 0.1 mg/L Sch-B reduced the level of MDA and increased the cleavage rate, blastocyst rate, and total cell number of blastocysts compared to the rate and number in the control group. However, no significant difference was observed in the ROS levels of blastocysts between the treatment and the control groups. The expression levels of CAT, SOD2, and Bcl-2 in IVF-blastocysts formed using sperm stored for one day at 17 °C were significantly higher than those in the control blastocysts. On day 4 of storage, CAT and Bcl-2 expression were significantly higher in IVF-blastocysts formed from sperm treated with 0.1 mg/L Sch-B than that in the control blastocysts. The ratio of Bax/Bcl-2 was also significantly higher in IVF-blastocysts formed using Sch-B-treated sperm. Our findings demonstrate that treatment with Sch-B can protect boar sperm from oxidative stress during liquid preservation and can increase the fertilization ability of the sperm. • Schisandrin B can significantly improve the quality of boar sperm during long-term storage. • Schisandrin B protects boar sperm stored at 17 °C by against oxidative damage. • Schisandrin B treatment of boar sperm improves fertilization rate and quality of the embryo. [ABSTRACT FROM AUTHOR]

Copyright of Indonesian Journal of Animal & Veterinary Sciences / Jurnal Ilmu Ternak dan Veteriner is the property of Indonesian Center for Animal Science Research & Development and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Mammalian spermatozoa are highly energized cells in which most of the proteins and activated signaling cascades are involved in the metabolic pathways. Flavin adenine dinucleotide (FAD) has one of the most important roles in the correct functional activity of spermatozoa since it acts as a cofactor for flavoenzymes, critical for proper metabolism and predominantly located in mitochondria. Non-invasive, vital and non-traumatic examination of sperm FAD level and microenvironment could be performed by fluorescence lifetime imaging microscopy (FLIM). In this study, we assessed the metabolic status of spermatozoa from healthy donors and found that FLIM could be used to segregate and separate the male germ cells according to the type of metabolic activity which corresponds with spermatozoa motility measured in standard spermogram tests. [Display omitted] • FLIM parameters reveal the heterogeneity of the sperm sample. • The Type of sperm motility are associated with FLIM parameters. • FLIM technology could be used in ART centers due to the non-invasive procedure and gentle effect on the cell. [ABSTRACT FROM AUTHOR]

Appropriate fertilization practices play a crucial role in the maintenance of genetic diversity within captive fish broodstocks. We assessed the influence of in vitro storage of gametes (0-, 7- and 14-days post-stripping) at 1 °C on ovarian fluid pH, sperm motility, and embryo viability in the critically endangered Finnish landlocked salmon (Salmo salar m. sebago) population. A repeated full-factorial design used in both sperm-ovarian fluid activation analyses and fertilizations allowed us to separate the effects of female, male and their interaction on gamete quality and embryo survival across storage times. The ovarian fluid pH decreased most significantly during the first week of storage and showed considerable variation among females. The decrease of sperm motility varied across males, females and male–female (sperm-ovarian fluid) combinations. The mean proportion of viable embryos was initially 96.4%, but dropped to 45.5% after 7 days, and to 13.1% after 14 days of gamete storage. Embryo viability responses to gamete storage time predominantly increased among- and within-female variation but also showed major differences between female × male combinations (non-additive genetic effects). Interestingly, embryo viability showed virtually complete re-ranking (change of order) among full-sib families between the first two fertilization times (rs = − 0.03) but consistency between the first and last times (rs = 0.59). Overall, our results indicate that even when the primary aim is to maximize the genetic diversity of endangered fish populations, it may be advisable to prioritize minimal egg storage time rather than maximizing the number of parental combinations in fertilizations. [ABSTRACT FROM AUTHOR]

Purpose: To determine if creating voting ensembles combining convolutional neural networks (CNN), support vector machine (SVM), and multi-layer neural networks (NN) alongside clinical parameters improves the accuracy of artificial intelligence (AI) as a non-invasive method for predicting aneuploidy. Methods: A cohort of 699 day 5 PGT-A tested blastocysts was used to train, validate, and test a CNN to classify embryos as euploid/aneuploid. All embryos were analyzed using a modified FAST-SeqS next-generation sequencing method. Patient characteristics such as maternal age, AMH level, paternal sperm quality, and total number of normally fertilized (2PN) embryos were processed using SVM and NN. To improve model performance, we created voting ensembles using CNN, SVM, and NN to combine our imaging data with clinical parameter variations. Statistical significance was evaluated with a one-sample t-test with 2 degrees of freedom. Results: When assessing blastocyst images alone, the CNN test accuracy was 61.2% (± 1.32% SEM, n = 3 models) in correctly classifying euploid/aneuploid embryos (n = 140 embryos). When the best CNN model was assessed as a voting ensemble, the test accuracy improved to 65.0% (AMH; p = 0.1), 66.4% (maternal age; p = 0.06), 65.7% (maternal age, AMH; p = 0.08), 66.4% (maternal age, AMH, number of 2PNs; p = 0.06), and 71.4% (maternal age, AMH, number of 2PNs, sperm quality; p = 0.02) (n = 140 embryos). Conclusions: By combining CNNs with patient characteristics, voting ensembles can be created to improve the accuracy of classifying embryos as euploid/aneuploid from CNN alone, allowing for AI to serve as a potential non-invasive method to aid in karyotype screening and selection of embryos. [ABSTRACT FROM AUTHOR]

This study was designed to evaluate the protective potentials of chitosan nanoparticles (ChNPs) against silver nanoparticle (AgNP)-induced reproductive toxicity in male Wister albino rats. AgNPs, ChNPs, and AgNPs particles coated with ChNPs were characterized by using transmission electron microscope. Control rats were injected interperitoneally with 0.5% aqueous carboxymethyl cellulose. Second group was given ChNPs at a dose 300 mg/kg bwt. Third group was given AgNPs at a dose 50 mg/kg bwt. Fourth group was given AgNPs with chitosan nanoparticles simultaneously. Fifth group was given silver nanoparticles coated with chitosan nanoparticles at a dose 300 mg/kg bwt. TEM showed the formation of AgNPs with average size of 42.7 nm, ChNPs with average size of 33.3 nm, and AgNPs coated with ChNPs with average size of 48.1 nm. AgNPs significantly reduced serum levels of FSH, LH, testosterone and prolactin, sperm count, morphology index, vitality, total motility and progressive motility, the activities of catalase and superoxide dismutase, and the concentration of reduced glutathione in testicular tissues. However, it significantly increased malondialdehyde concentration in testicular tissues, sperm abnormalities, testicular tissue damages, non-progressive motility, and immotile sperms. On the contrast, ChNPs ameliorated AgNP-induced alteration in serum levels of sex hormones, spermogram, and testicular tissue's structure and functions. These results indicated that ChNPs had protective potential against AgNP-induced reproductive toxicity and ChNPs coating AgNPs had more potent protective effect than ChNPs administrated together with AgNPs. [ABSTRACT FROM AUTHOR]

Perfluorooctanesulfonate acid (PFOS) is a typical persistent organic pollutant that widely exists in the environment. To clarify the toxic effects and mechanisms of PFOS and to find effective intervention strategies have been attracted global attention. Here, we investigated the effects of PFOS on the male reproductive system and explored the potential protective role of 1α,25‐dihydroxyvitamin D3 (1α,25(OH)2D3). Our results showed that 1α,25(OH)2D3 intervention significantly improved PFOS‐induced sperm quality decline and testicular damage. Moreover, 1α,25(OH)2D3 aggrandized the total antioxidant capacity. Furthermore, after PFOS exposure, the transcription factor nuclear factor erythroid‐related factor 2 (Nrf2) was adaptively increased together with its target genes, such as HO‐1, NQO1, and SOD2. Meanwhile, 1α,25(OH)2D3 ameliorated PFOS‐induced augment of Nrf2 and target genes. These findings indicated that 1α,25(OH)2D3 might attenuate PFOS‐induced reproductive injury in male mice via Nrf2‐mediated oxidative stress. [ABSTRACT FROM AUTHOR]

Recently published single-cell sequencing data from individual human sperm (n=41,189; 969-3377 cells from each of 25 donors) offer an opportunity to investigate questions of inheritance with improved statistical power, but require new methods tailored to these extremely low-coverage data (~0.01× per cell). To this end, we developed a method, named rhapsodi, that leverages sparse gamete genotype data to phase the diploid genomes of the donor individuals, impute missing gamete genotypes, and discover meiotic recombination breakpoints, benchmarking its performance across a wide range of study designs. We then applied rhapsodi to the sperm sequencing data to investigate adherence to Mendel's Law of Segregation, which states that the offspring of a diploid, heterozygous parent will inherit either allele with equal probability. While the vast majority of loci adhere to this rule, research in model and non-model organisms has uncovered numerous exceptions whereby 'selfish' alleles are disproportionately transmitted to the next generation. Evidence of such 'transmission distortion' (TD) in humans remains equivocal in part because scans of human pedigrees have been under-powered to detect small effects. After applying rhapsodi to the sperm data and scanning for evidence of TD, our results exhibited close concordance with binomial expectations under balanced transmission. Together, our work demonstrates that rhapsodi can facilitate novel uses of inferred genotype data and meiotic recombination events, while offering a powerful quantitative framework for testing for TD in other cohorts and study systems. [ABSTRACT FROM AUTHOR]

Objectives: In recent years, smoking water pipes or hookah has increased among adolescents in most countries. Although there is evidence in support of the negative effects of this type of smoking on human health, such as the increased risk of lung disease, little is known about the potential effects of hookah smoking on the male reproductive system, especially on the molecular aspects of sperm. Patients and methods: This cross-sectional study examined sperm DNA fragmentation index, protamine 1 and 2 (PRM1 and PRM2) genes expression, and oxidant status in normozoospermic hookah smokers in comparison with non-smoker controls. Results: Our results showed significantly higher rates of DNA fragmentation, protamine deficiency, and abnormal chromatin condensation in the spermatozoa of hookah smokers (P <.0001). Also, protamine gene expression showed a remarkable decrease in hookah smokers (1.55 ± 2.54 and 0.33 ± 0.54) compared to the controls (3.49 ± 5.41 and 1.22 ± 1.96), although the reduction was not statistically significant (P =.155 and P =.066, respectively). Moreover, a significantly higher level of semen MDA was observed in the case group compared to the controls (0.39 ± 1.04 vs 0.15 ± 0.21; P =.013). Conclusion: According to our study, although hookah smoking does not have a significant effect on sperm parameters, it may have deleterious effects on DNA integrity, oxidative status, and nuclear protein levels of spermatozoa. [ABSTRACT FROM AUTHOR]

Objective: We aimed to evaluate intracytoplasmic sperm injection (ICSI) outcomes in young patients with diminished ovarian reserve (DOR) plus severe male factor (SMF) compared with age‐matched controls with DOR. Study design: A total of 189 infertile women under 35 years with DOR undergoing ICSI procedures were included retrospectively. Participants whose partners' sperm analysis was normal considered as the DOR group (n = 154) and whose partners' had SMF considered as the DOR + SMF group (n = 35). The two groups were compared regarding cycle characteristics and pregnancy outcomes. Results: Demographic features except infertility duration were similar between two groups. The duration of infertility was significantly longer in the DOR + SMF group compared to the DOR group (p = 0.02). Ovarian stimulation characteristics, oocyte retrieval parameters, fertilization rate, quality of embryos, embryo cancellation rate, and development up to blastocyst stage were found similar between two groups. Implantation, clinical pregnancy, abortion, and live birth rate, multiple pregnancy rate per cycle were distributed homogenously between the DOR and DOR + SMF groups. Regarding perinatal and neonatal outcomes of groups, fetal height and weight were significantly lower in DOR + SMF group than in DOR group (p = 0.001 and 0.01, respectively). Gestational week at delivery was lower in the DOR + SMF group compared to the DOR group (p < 0.0001). Conclusion: Fetal anthropometric measures were lower regarding to preterm delivery in the DOR + SMF group than the DOR group. Large sample‐sized studies should be performed to explain the decreased gestational week at the time of delivery in the DOR + SMF group. [ABSTRACT FROM AUTHOR]

Cattleyaks are equally adaptable to harsh environment as yaks, but produce far more milk and meat in terms of quality and quantity. However, male cattleyaks with active secondary sexuality are infertile and have restricted productivity and breeding of yaks. Much researches continue to be done in regard to the differences in transcriptome profiling in cattleyak epididymis with respect to yak epididymis. The caput segment of the epididymis is highly specialized for the initiation of spermatozoa maturation, synthesis and secretion. We used RNA-Seq technology to comparatively analyze differentially expressed genes (DEGs) associated with sperm maturation between the caput epididymis of yak and cattleyak. Transcriptomic profiling identified 109 DEGs in which 44 were upregulated and 65 were downregulated. 8 DEGs were validated by quantitative real-time PCR. DEGs were analyzed by GO and KEGG analysis to screen the key genes involved in sperm maturation. The upregulation of PAOX and ATP2C2 may be associated with toxicity and apoptosis resistance in cattleyak with respect to yak. However, downregulated DEFB109, DEFB121 , DEFB123 , DEFA1 , LY6G5C, SLC13A2 , CST3, CRYBA4 and ADAM28 were associated with innate immune response, sperm maturation, motility and antimicrobial functions. AMPK and Hedgehog signaling pathways were involved in the top-listed five significantly enriched pathways, and the downregulation of HNF4α and LRP2 may have contributed to infertility in cattleyak. The data provide a powerful resource, contributing to the knowledge on the molecular mechanisms underlying male cattleyak infertility. • RNA-Seq technology comparative analysis of DEGs in sperm maturation between the caput epididymis of yak and cattleyak. • Transcriptomic profiling identified 109 DEGs, 44 upregulated and 65 downregulated. • Upregulation of PAOX and ATP2C2 may cause toxicity and apoptosis resistance in cattleyak with respect to yak. Downregulation of HNF4α and LRP2 may cause infertility in cattleyak. [ABSTRACT FROM AUTHOR]

Heat stress (HS) is the most substantial environmental issue in rabbit health status impairment, which can lower productivity. Maca is a common medicinal plant with important biological activities including antioxidant, anti‐inflammatory, anti‐fatigue, and neuroprotective effects. The present research explored the alleged protective role of Maca extract in alleviating the adverse impact of HS on rabbits. Growing V‐line 6‐week‐old buck rabbits (N = 48) were orally administered Maca extract at levels of 0 (MAC0), 200 (MAC2), 400 (MAC4), or 600 (MAC6) mg/head twice per week. Compared to the control groups, rabbits provided Maca had better growth performance, feed intake and feed conversion ratio. Liver and tests weights were significantly higher (p < 0.05) in Maca treated groups compared to the control. hemoglobin, white blood cell, red blood corpuscles, platelet count, hematocrit, and lymphocytes were significantly increased in the MAC4 group compared with the control group. MAC4 treatment significantly reduced aspartate aminotransferase and malondialdehyde levels compared with other groups. Moreover, the concentrations of total bilirubin, creatinine, glucose, total cholesterol, and triglycerides were lower (p < 0.05) in the MAC4 group than in the controls. Oral administration of Maca improved cortisol and testosterone values, total antioxidant capacity, and superoxide dismutase. The MAC4 group exhibited significant improvement (p < 0.05) in sperm motility, survival, membrane functionality, concentration, and libido, with a significant decrease in abnormal sperm compared with the control group. Also, aforementioned group showed sperm cells with a normal structure and intact plasma membranes, acrosomes, and well‐organised axonemal components. Histopathological screening of liver sections showed moderate to severe degenerative and necrotic changes in rabbits exposed to HS, which were alleviated with Maca oral administration. In conclusion, oral administration of Maca extract at 400 or 600 mg/head‐weekly enhanced growth performance, hemato‐biochemical attributes, antioxidant status, and semen quality in rabbits during the hot season. [ABSTRACT FROM AUTHOR]

In this paper are presented the experimental results obtained after the administration of polyphenols of plant origin extracted from green walnuts and their influence on the ratio of free amino acids content in blood serum and reproductive material of roosters. It was demonstrated that the total content of free amino acids in the blood serum of roosters is 437.14±45.23 mcm/100 ml. The highest share belongs to proline – 31.34%, glycine – 10.63%, alanine – 7.73%, threonine – 7.62% and glutamine – 7.37% of the total content of all amino acids. Seminal fluid is characterized by a 3 times higher content of free amino acids than in blood serum, which constitutes - 1885.42±130.43 mcm/100 ml. For seminal fluid, it is characteristic that the highest share is represented by the content of asparagine – 16.3%, glutamine 40.16%, glutamic acid – 9.64% of the summary content of amino acids in the seminal plasma. The total content of free amino acids in the reproductive cells of roosters constitutes 110.69±3.28 mcm/100 ml. In this biological material the highest share belongs to taurine with a value of 37.28%, followed by arginine with a value of 17.68%, glycine – 15.19%, alanine – 6.55%, proline – 6.43%, glutamine – 5.80%. [ABSTRACT FROM AUTHOR]

Simple Summary: Both in human medicine and in the veterinary field, lipidomic techniques have been applied extensively in order to evaluate health status. In equine species where the subject of focus is the fertility and sperm quality of the equids, lipidomic assessment can be a useful tool to better understand the metabolic transformation of the fatty acids of the spermatozoa as well as the erythrocyte membrane and their role on spermiogenesis. The purpose of this study is to evaluate the composition of the spermatozoa's membranes of 26 healthy male Martina Franca donkeys and the possible correlation with the sperm parameters and the erythrocytes' membrane. The results demonstrate that the membrane of erythrocytes and spermatozoa are different, which provides a deeper understanding of the distribution and the role of these fatty acids in sperm parameters that are highly associated with fertility. Fatty acid-based lipidomic analysis has been widely used to evaluate health status in human medicine as well as in the veterinary field. In equine species, there has been a developing interest in fertility and sperm quality. Fatty acids, being the principal components of the membranes, play an active role in the regulation of the metabolic activities, and their role on spermiogenesis seems to be of great importance for the resulting quality of the sperm and, thus, fertility. With the application of widely used lipidomic techniques, the aim of this study was to evaluate: (a) the fatty acid content of the spermatozoa's membranes of 26 healthy male Martina Franca donkeys and its possible correlation with sperm parameters, and (b) the evaluation of the composition of the red blood cells' membrane. PUFA omega-6 are the principal components (40.38%) of the total PUFA content (47.79%) in both types of cells; however, DPA is the predominant one on the spermatozoa's membrane (27.57%) but is not present in the erythrocyte's membrane. Spermatozoa's motility (%) is positively correlated with stearic acid and EPA, and progressive motility (%), with oleic acid. These findings offer information on the composition of both types of cells' membranes in healthy male MF donkeys and reflect the metabolic transformations of the spermatozoa's membrane during the maturation period, providing a better perception of the role of fatty acids in sperm parameters and fertility. [ABSTRACT FROM AUTHOR]