The Effect of Aqueous Lessertia frutescens Extract on TM3 Leydig Cells Exposed to TNF-α in vitro.

BACKGROUND: Extractions of Lessertia frutescens (Lf) are shown to have immune modulation, anti-inflammatory and antioxidant properties. However, Lf is also cytotoxic, antiproliferative, and pro-apoptotic in vitro. Furthermore, Lf extractions may influence steroidogenesis. Nevertheless, the impact on Leydig cell function has not previously been investigated. As tumor necrosis factor-alpha (TNF-α) is known to cause Leydig cell dysfunction under inflammatory conditions, it is further proposed that Lf extracts may protect against the negative impact of TNF-α on Leydig cells. The aim of the study was to investigate the effect of an aqueous Lessertia frutescens extract (LFE) on Leydig cells exposed to TNF-αin vitro. METHODS: Human chorionic gonadotrophin-stimulated TM3 Leydig cells were exposed for 24 h to (a) TNF-α (0.1, 1, 10, 100 ng/mL), (b) LFE (0.01, 0.1, 1, 10, 100 ng/mL), and (c) co-exposure to 10 ng/mL TNF-α and LFE (0.01, 0.1, 1, 10, 100 ng/mL). We analyzed cell viability, cytotoxicity, caspase 3/7 activation, testosterone concentration, and intracellular superoxide. RESULTS: TNF-α exposure decreased cell viability, increased cytotoxicity, and caspase 3/7, with no significant effect on intracellular superoxide in TM3 Leydig cells. When LFE concentrations of 0.01-10 ng/mL were tested, we observed improved vitality and reduced levels of caspase 3/7. At 100 ng/mL, LFE decreased viability and increased cytotoxicity and caspase 3/7. However, LFE did not affect intracellular superoxide. Furthermore, LFE protected against 10 ng/mL TNF-α-induced cytotoxicity and apoptosis, except at the highest concentration. LFE alone and in co-culture with 10 ng/mL TNF-α increased testosterone at high concentrations. CONCLUSIONS: In our TM3 Leydig cell model, LFE protected against TNF-α-induced cytotoxicity and early apoptosis, except at the highest experimental concentrations, where it was cytotoxic. These effects were not mediated through a change in intracellular superoxide. Although further investigations are warranted, aqueous LFE may protect against TNF-α-induced Leydig cell dysfunction.

Highly Cited Articles in the Field of Male Infertility and Antioxidants: A Scientometric Analysis.

PURPOSE: The objective of this scientometric analysis was to recognize the top 100 cited articles on 'Male infertility and Antioxidants' and analyze its publication characteristics. MATERIALS AND METHODS: The Scopus database was used to retrieve related articles and the top 100 identified based on citation rate. RESULTS: The articles were published in 56 journals between 1995 and 2019 with a median (interquartile range) citation score of 17 (5-62). Among the top 100 articles, 69 were clinical studies, which included controlled and blinded (33.33%), prospective (27.54%), randomized-controlled trials (26.09%), uncontrolled (11.59%), and retrospective (1.45%) studies. In addition to conventional semen parameters, advanced sperm function tests such as oxidative stress (51%) and sperm DNA damage (23%) were reported. Pregnancy rate (33%) was found to be the most reported reproductive outcome. Antioxidant therapy was mostly investigated in male cohorts with sperm abnormalities such as asthenozoospermia (28%) and clinical conditions such as idiopathic male infertility (20%), varicocele/varicocelectomy (17%) and general male infertility (16%). CONCLUSIONS: The most influential publications on antioxidants and male infertility were identified for the first time in the literature. This will serve as a reliable source of information for researchers and clinicians alike.

Diagnostic value of advanced semen analysis in evaluation of male infertility.

Conventional semen analysis is the standard of care to initially evaluate the fertility status of a male patient. However, it has some limitations and among these are failure to correctly identify the aetiology underlying fertility problems, intra- and inter-observer variability and incomplete information about sperm function. Considering these drawbacks, advanced semen tests have been developed to assess male infertility, including sperm function tests, oxidative stress (OS) and sperm DNA fragmentation (SDF) tests. This review illustrates the commonly utilised sperm function techniques, along with the assays used to assess SDF and OS and their diagnostic value.

A scientometric analysis of research publications on male infertility and assisted reproductive technology.

Assisted reproductive technologies (ART) are considered as one of the primary management options to address severe male factor infertility. The purpose of this study was to identify the research trends in the field of male infertility and ART over the past 20 years (2000-2019) by analysing scientometric data (the number of publications per year, authors, author affiliations, journals, countries, type of documents, subject area and number of citations) retrieved using the Scopus database. We used VOS viewer software to generate a network map on international collaborations as well as a heat map of the top scientists in this field. Our results revealed a total of 2,148 publications during this period with Cleveland Clinic Foundation contributing the most (n = 69). The current scientometric analysis showed that the research trend on ART has been stable over the past two decades. Further in-depth analysis revealed that density gradient centrifugation (46%) and intracytoplasmic sperm injection (59.2%) are the most reported techniques for sperm separation and ART, respectively. Additionally, azoospermia was the most studied clinical scenario (60.6%), with majority of articles reporting pregnancy rate (47.25%) as the primary reproductive outcome for ART. This study provides insight into the current focus of research in the area of male infertility and ART as well as the areas that require further research in future.

The Impact of Single- and Double-Strand DNA Breaks in Human Spermatozoa on Assisted Reproduction.

Several cellular insults can result in sperm DNA fragmentation either on one or both DNA strands. Oxidative damage, premature interruption of the apoptotic process and defects in DNA compaction during spermatogenesis are the main mechanisms that cause DNA breaks in sperm. The two-tailed Comet assay is the only technique that can differentiate single- (SSBs) from double- (DSBs) strand DNA breaks. Increased levels of the phosphorylated isoform of the H2AX histone are directly correlated with DSBs and proposed as a molecular biomarker of DSBs. We have carried out a narrative review on the etiologies associated with SSBs and DSBs in sperm DNA, their association with reproductive outcomes and the mechanisms involved in their repair. Evidence suggests a stronger negative impact of DSBs on reproductive outcomes (fertilization, implantation, miscarriage, pregnancy, and live birth rates) than SSBs, which can be partially overcome by using intracytoplasmic sperm injection (ICSI). In sperm, SSBs are irreversible, whereas DSBs can be repaired by homologous recombination, non-homologous end joining (NHEJ) and alternative NHEJ pathways. Although few studies have been published, further research is warranted to provide a better understanding of the differential effects of sperm SSBs and DSBs on reproductive outcomes as well as the prognostic relevance of DNA breaks discrimination in clinical practice.

Unraveling the Footsteps of Proteomics in Male Reproductive Research: A Scientometric Approach.

Significance: Male reproductive research at molecular level has gained more attention as it offers the cellular mechanisms and biological pathways implicated in the reproductive physiology. Several researchers across the world have used global proteomic approach in conjunction with advanced bioinformatics software to identify putative biomarkers for various male infertility conditions. Recent Advances: Introduction of advance proteomic platforms has made it easier to generate enormous amount of data in a short period of time. In this article, we have reviewed the functional and comparative proteomic studies in the area of male reproductive research. We have discussed the key proteins and associated cellular pathways such as oxidative phosphorylation and mitochondrial dysfunction implicated in the various male infertility conditions. Furthermore, for the first time scientometric approach was used to analyze the publication trends and hot topics in proteomics of male reproductive research. Critical Issues: Analysis of publication trends revealed that majority of the published studies were focused on varicocele and asthenozoospermia, while very limited research has been conducted on assisted reproductive technology (ART). This area of research requires more attention as it would facilitate identification of novel biomarkers to catalogue proteomic characteristics of spermatozoa for achieving better results in ART. Future Directions: Future research should be focused on the development and validation of a biomarker panel for specific male infertility scenarios based on etiology. Translation of validated proteomic biomarkers into tests or assays for male infertility conditions would enable the physician to provide better management for the patients.