Efficacy of combined antioxidant therapy in male subfertility-A systematic review and meta-analysis

Antioxidant therapy is a potentially promising approach to improve clinical outcomes for couples undergoing assisted reproduction techniques long-term. The review aims to (a) collate evidence for the effectiveness of combined oral antioxidant supplementation, including a head-to-head comparison in the treatment of male subfertility, and (b) investigate whether other intervention features, including duration, specific combinations, or dosage affect clinical outcomes in this population. Randomized controlled trials (RCTs) that examined the effectiveness of combined antioxidants on male subfertility, electronic databases including PubMed, Embase, CINAHL, PSYCHINFO, and Cochrane Central Register of Controlled Trials (CENTRAL) were searched. We also searched for unpublished data and references of identified articles. Two reviewers screened eligible studies according to pre-defined criteria and relevant data extracted. The Jadad scale assessed the quality of studies. The study used RevMan version 5.4.1 Meta-analysis software to analyze the effect of combination antioxidants for each outcome measure. Metadata was presented as relative risks for dichotomous outcomes and as standardized mean differences (or mean differences) for continuous outcomes. The systematic review and meta-analysis aimed to report available evidence of whether combination antioxidant is effective and safe in sub-fertile men undergoing assisted reproductive techniques. Also, this review highlighted whether any specific oral antioxidant combinations, dosage, or duration of therapy have a major influence on the clinical outcomes.

Male Subfertility; Challenges and advancement in the andrology clinic at Khyber Pakhtunkhwa, Pakistan

Objective: This study is aimed to discuss the challenges in dealing the infertile male and advances in the treatment of male infertility. Material and methods: The study included infertile male patients who presented to andrology outpatient as primary or secondary infertility between December 2018 and January 2021. The data detailed different aspects of challenges and advances in male infertility treatment. The data analysisone with SPSS. Results: Total 289 patients included, most of them (74%) presented as primary infertility and a quarter presented as secondary infertility. The mean delay in presentation was 6.8 years which were due to treatment from non-andrologist doctors of different specialities (53.9%), hakims (15.2%), quacks (13.8%), gynaecologists (10.3%) and some were reluctant to tell their problem (6.5%). The diagnosis was N.O.A (42.9%), unexplained infertility (24.2%), varicocele (22.8%), OA (6.2%), OAT syndrome (2.7%) and CABVD (1%). Different treatment option opted were vasography plus vasovasostomy or vasoepididmostomy (31.1%), ART (23.9%), MSV (22.8%) and medical treatment (22.1%). Vasography plus vasovasostomy or vasoepididmostomy and medical treatment were the available options provided. There was no ART facility and those who were counseled for referral either their unwillingness or cost resulted in a hurdle in their provision. Conclusion: There are still a number of challenges in treating infertile men. Recently provision of medical and microsurgical treatment at andrology clinic resulted in proper treatment of a large number of infertile men who previously received treatment from un- related facilities.

Subfertility in young male mice mutant for chromatin remodeler CECR2

Defects in spermatogenesis are an important cause of male infertility. Multiple aspects of spermatogenesis are controlled by chromatin remodelers, including regulating transcription. We previously described mutations in chromatin remodeling gene Cecr2 that resulted in the lethal neural tube defect exencephaly in most mutant mice, and subfertility in mice that were non-penetrant for exencephaly. Here, we show that the severity of male subfertility is dependent on age. Cecr2GT/Del males contain two mutant alleles, one of which is hypomorphic and therefore produces a small amount of protein. These males sire the fewest pups just after sexual maturity (88% fewer than Cecr2+/+ at P42-60) but improve with age (49% fewer than Cecr2+/+ at P81-100), although never completely recovering to Cecr2+/+ (wild type) levels. When young, they also have defects in testis histology, in vivo fertilization frequency, sperm number and motility, and testis weight that show similar improvement with age. Immunostaining of staged seminiferous tubules showed CECR2 in type A, In and B spermatogonia, and less in preleptotene and leptotene spermatocytes. Histological defects were first apparent in Cecr2GT/Del testes at P24, and RNA-seq analysis revealed 387 differentially expressed genes. This included 66 genes on the X chromosome (almost double the number on any other chromosome), all more highly expressed in Cecr2GT/Del testes. This inappropriate expression of X chromosome genes could be caused by a failure of effective meiotic sex chromosome inactivation. We identify several abnormally expressed genes that may contribute to defects in spermatogenesis at P24. Our results support a role for Cecr2 in juvenile spermatogenesis.

Diet-induced- and genetic- obesity differentially alters male germline histones

Obesity, an established risk factor for male subfertility or infertility, is primarily due to genetic and environmental causes. Our earlier studies have shown differential effects of high fat diet-induced- (DIO) and genetically inherited- (GIO) obesity on DNA methylation in male germline and its subsequent effect on fertility. Here, we hypothesized that the effects of DIO and GIO on histone modifications in male germline could also contribute to fertility defects. We observed that DIO affected both active (H3K4me3, H3ac, and H4ac) and repressive (H3K9me3 and H3K27me3) histone marks in testis and their cell types, whereas GIO solely altered acetylated histones. This correlated with deregulation of histone-modifying enzymes in testis of both obese groups. Further, we also observed a decrease in chromatin remodelers in testis of DIO group, which were increased in GIO group. Besides, there was an increase in core histones and a decrease in histone marks along with protamine deficiency in spermatozoa of DIO group, whereas only H3K4me3 levels were increased in spermatozoa of GIO group. Moreover, we observed alterations in the expression and enrichment patterns of a few developmental genes harbored by the active histone mark in resorbed embryos sired by the DIO rats. Together these epigenetic defects in male germline could alter sperm quality and cause fertility defects in these obese groups. Differential changes in two obese groups could also be attributed to differences in their pathophysiological variations. Our study highlights epigenetic differences between DIO and GIO in male germline and its subsequent impact on male fertility.

Accumulation of Seminolipid in Sertoli Cells Is Associated with Increased Levels of Reactive Oxygen Species and Male Subfertility: Studies in Aging Arsa Null Male Mice

Seminolipid (also known as sulfogalactosylglycerolipid-SGG), present selectively in male germ cells, plays important roles in spermatogenesis and sperm–egg interaction. The proper degradation of SGG in apoptotic germ cells is also as important. Sertoli cells first phagocytose apoptotic germ cells, then Sertoli lysosomal arylsulfatase A (ARSA) desulfates SGG, the first step of SGG degradation. We have reported that aging male Arsa−/− mice become subfertile with SGG accumulation in Sertoli cell lysosomes, typical of a lysosomal storage disorder (LSD). Since reactive oxygen species (ROS) levels are increased in other glycolipid-accumulated LSDs, we quantified ROS in Arsa−/− Sertoli cells. Our analyses indicated increases in superoxide and H2O2 in Arsa−/− Sertoli cells with elevated apoptosis rates, relative to WT counterparts. Excess H2O2 from Arsa−/− Sertoli cells could travel into testicular germ cells (TGCs) to induce ROS production. Our results indeed indicated higher superoxide levels in Arsa−/− TGCs, compared with WT TGCs. Increased ROS levels in Arsa−/− Sertoli cells and TGCs likely caused the decrease in spermatogenesis and increased the abnormal sperm population in aging Arsa−/− mice, including the 50% decrease in sperm SGG with egg binding ability. In summary, our study indicated that increased ROS production was the mechanism through which subfertility manifested following SGG accumulation in Sertoli cells.

First insights on seminal extracellular vesicles in chickens of contrasted fertility

Male subfertility causes are very varied and sometimes related to post-gonadic maturation disruption, involving seminal plasma constituents. Among them, extracellular vesicles are involved in key exchanges with sperm in mammals. However, in birds, the existence of seminal extracellular vesicles is still debated. The aim of the present work was first to clarify the putative presence of extracellular vesicles in the seminal plasma of chickens, secondly to characterize their size and protein markers in animals showing different fertility, and finally to make preliminary evaluations of their interactions with sperm. We successfully isolated extracellular vesicles from seminal plasma of males showing the highest differences in semen quality and fertility by using ultracentrifugation protocol (pool of 3 ejaculates/rooster, n =3/condition). Size characterization performed by electron microscopy revealed a high proportion of small extracellular vesicles (probably exosomes) in chicken seminal plasma. Smaller extracellular vesicles appeared more abundant in fertile than in subfertile roosters, with a mean diameter of 65.12 and 77.18 nm, respectively. Different protein markers of extracellular vesicles were found by western blotting (n = 6/condition). Among them, HSP90A was significantly more abundant in fertile than in subfertile males. In co-incubation experiments (n = 3/condition), extracellular vesicles enriched seminal fractions of fertile males showed a higher capacity to be incorporated into fertile than into subfertile sperm. Sperm viability and motility were impacted by the presence of extracellular vesicles from fertile males. In conclusion, we successfully demonstrated the presence of extracellular vesicles in chicken seminal plasma, with differential size, protein markers and putative incorporation capacity according to male fertility status.

Male Infertility - An Analysis in a Low Ressource Setting

Purpose The prevalence of male subfertility is increasing among couples seeking fecundity in recent years. A prospective study was carried out in a gynaecological outpatient , to determine the benefit of a low cost algorithm for diagnosis of adult males with suboptimal semen parameters. Materials and Methods Twenty nine males, who had a sperm concentration less than 15 million/ml were considered in the study. The cost of diagnosis was kept low ; not exceeding 150 US dollars. A similar algorithm of investigation was followed for all patients and medical treatment provided as per treatment guidelines .Results A confirmed etiology could be determined in 9 patients , while a probable diagnosis was suggested in 17 males. Endocrino-metabolic causes were the most important etiological factors in our Indian population . Lifestyle diseases like obesity , insulin resistance were prominently found in the suspected acquired hypogonadotropic hypogonadism group..Medical treatment was provided to 24 males, and 64% showed good improvement in seminal count , after 3 months of therapy. Only selected patients were referred to higher teritiary centres, for specific interventions like sperm retrieval techniques , intracytoplasmic sperm injection of the female partner. Conclusion This study proves that male subfertility can be managed in a low resource semiurban centre, with the help of a simple algorithm with ease and proficiency,hence reducing economic burden on these couples. Also, this analysis has helped us to ascertain the relative propensity of etiological factors among adult males with infertility ,in North India in the present time.

Deactivation of the JNK Pathway by GSTP1 Is Essential to Maintain Sperm Functionality

Fifty percent of male subfertility diagnosis is idiopathic and is usually associated with genetic abnormalities or protein dysfunction, which are not detectable through the conventional spermiogram. Glutathione S-transferases (GSTs) are antioxidant enzymes essential for preserving sperm function and maintaining fertilizing ability. However, while the role of GSTP1 in cell signaling regulation via the inhibition of c-Jun N-terminal kinases (JNK) has been enlightened in somatic cells, it has never been investigated in mammalian spermatozoa. In this regard, a comprehensive approach through immunoblotting, immunofluorescence, computer-assisted sperm assessment (CASA), and flow cytometry analysis was used to characterize the molecular role of the GSTP1–JNK heterocomplex in sperm physiology, using the pig as a model. Immunological assessments confirmed the presence and localization of GSTP1 in sperm cells. The pharmacological dissociation of the GSTP1–JNK heterocomplex resulted in the activation of JNK, which led to a significant decrease in sperm viability, motility, mitochondrial activity, and plasma membrane stability, as well as to an increase of intracellular superoxides. No effects in intracellular calcium levels and acrosome membrane integrity were observed. In conclusion, the present work has demonstrated, for the first time, the essential role of GSTP1 in deactivating JNK, which is crucial to maintain sperm function and has also set the grounds to understand the relevance of the GSTP1–JNK heterocomplex for the regulation of mammalian sperm physiology.