Andrologia

The present study aimed to determine the cryoprotective effect of melatonin supplements on the extender on the quality of canine epididymal spermatozoa. Epididymis from 20 castrated dogs were minced and incubated in Tris buffer. Spermatozoa were diluted in tris-citric acid-fructose (TCF) extender with two doses of melatonin (0.002 and 0.0035 mol) as well as a control (0.0 mol). The samples were packed in 0.25 ml straws after 2 hr of equilibration at 5°C and kept in liquid nitrogen (−196°C). Sperm parameters such as viability, motility, integrity of acrosome, sperm membrane functioning, and DNA integrity were assessed after thawing (37°C for 30 s). In addition, lipid peroxidation was assessed using the determination of malondialdehyde (MDA) levels. Results revealed that adding 0.0035 mol melatonin to the cryopreservation medium enhanced () the measured parameters of spermatozoa compared to the control and 0.002 mol melatonin. Levels of MDA were obviously () lower when 0.0035 mol melatonin was added in comparison to 0.002 mol melatonin and the control. Eventually, adding 0.0035 mol melatonin to the TCF extender significantly improved the percentage of motility, viability, and integrity of both plasma membrane and acrosome integrity of dog epididymal spermatozoa. In addition, melatonin supplementation improved DNA integrity and reduced membrane lipid peroxidation.

Introduction. Valproic acid (VPA) is a widely used drug in the treatment of emotional and nervous depression, mental imbalances, epilepsy, and control of bipolar disorders. Despite its therapeutic effects, VPA has toxic effects on male reproduction. The testis is an organ sensitive to oxidative stress. On the other hand, thyme honey contains antioxidant compounds. The aim of this study was to investigate the effect of thyme honey on the histopathological changes of the testis in relation to the side effects of VPA in adult male Wistar rats. Methods. In this study, 48 Wistar rats were used, which were randomly divided into equal eight groups and received their desired substances. Including the control group, Th1, Th2, and Th3 groups receiving 1, 2, and 3 g/kg of thyme honey, respectively, VPA (500 mg/kg), VPA + Th1 (500 mg/kg of VPA + 1 g/kg of thyme honey), VPA + Th2 (500 mg/kg of VPA + 2 g/kg of thyme honey), and VPA + Th3 (500 mg/kg of VPA + 3 g/kg of thyme honey). After 60 days, evaluation of malondialdehyde (MDA), total antioxidant capacity (TAC), and histological studies of hematoxylin-eosin and Masson’s trichrome staining were performed. Results. The use of thyme honey significantly improved some parameters related to spermatogenesis, and in addition, it increased sperm production in the honey groups. In addition, the changes in MDA levels in the groups have shown a decrease in the amount of lipid peroxidation in the VPA + Th2 group. The amount of TAC also increased after increasing the dose of thyme honey. Conclusion. Based on the results of this study, it seems that thyme honey has been able to reduce the side effects of VPA to some extent due to its high percentage of antioxidants.

Background. Infertile men with nonobstructive azoospermia (NOA) associated with varicocele may benefit from varicocele repair with recovered sperms in semen. Currently, there is no clinically applied method to predict the success of seminal sperm recovery in this subset of azoospermic men. Objective. We aimed to evaluate the predictive role of the seminal testis expressed protein (TEX101) in sperm recovery in the semen of NOA men with palpable bilateral varicocele after microsurgical varicocelectomy. Patients and Methods. This prospective cross-sectional cohort study included 40 patients with NOA and bilateral varicocele. Seminal TEX101 levels were measured followed by subinguinal microsurgical varicocele repair. Two seminal analyses were carried out at 3- and 6-month follow-up periods to evaluate seminal sperm recovery. Results. After varicocele repair, sperms were recovered in the semen of 10/40 patients (25%) along the follow-up periods (seven patients after 3 months and additional three patients after 6 months). The preoperative median seminal TEX101 level was significantly higher in NOA men with seminal sperm recovery compared with NOA patients without seminal sperm recovery (13.5 vs. 9.8 ng/ml, ). Besides, the serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) median levels were significantly higher in NOA men without seminal sperm recovery compared with NOA men with seminal sperm recovery (, ). There were nonsignificant differences comparing the preoperative testicular volume or serum testosterone levels between the two investigated groups (, ). A cutoff value of 9.9 ng/ml was demonstrated to have preoperative TEX101 prediction of seminal sperm recovery (sensitivity of 90% and specificity of 57%). Conclusion. Preoperative seminal TEX101 could be considered as a biomarker for sperm recovery for seminal sperm recovery in NOA cases associated with varicocele after its surgical repair. Further work at this point with larger number of cases should be carried out to verify its potential importance.

Testicular germ cell tumors (TGCTs) are the most common solid malignant tumor in young men aged 20–34 years. Currently, the diagnosis and differentiation of TGCTs depend on immunohistochemical analysis, and the treatment methods are mainly surgery and chemoradiotherapy. Although immunotherapy has been applied in clinic, the response rate is not high, and also there is a lack of effective biomarkers. In this study, through high-throughput transcriptome sequencing and public data mining, the expression of hepatitis A virus cellular receptor 2 (HAVCR2) was found to be significantly upregulated in TGCT tissue and correlated with poor prognosis. Additionally, the expression level of HAVCR2 was negatively correlated with its DNA methylation but positively correlated with its copy number level. The HAVCR2 expression level was significantly positively correlated with the infiltration of six types of immune cells, including B cells, CD4+ T cells, CD8+ T cells, neutrophils, macrophages, and myeloid dendritic cells. Also, the higher its expression level, the higher the immune cell abundance. Tumor immune dysfunction and exclusion (TIDE) scoring analysis showed that HAVCR2 was related to the responsiveness to TGCT immunotherapy. Drug sensitivity analysis revealed that HAVCR2 was related to the sensitivity of multiple antitumor drugs. This study demonstrated that HAVCR2 has high potential as a biomarker for the diagnosis, prognosis, and treatment of TGCTs.

High altitude (HA) poses various dangers to living organisms, the most serious of which are oxidative stress and its accompanying metabolic problems, as well as hypoxia and its associated metabolic abnormalities. Acrylamide is a poisonous chemical produced under the oxidative stress as a result of intracellular reactive oxygen species formation and toxicity. Heating carbohydrates-rich meals give acrylamide that is widely used in the industry. The precise mechanism of acrylamide toxicity is unknown. The current study aimed to examine the impacts of Taify pomegranate juice (TPJ) from Taif area (Punica granatum L.) on acrylamide-induced testicular stress and dysfunction. Twenty four male of adult Wistar rats, were divided into four groups: Group 1 was a negative-control received only saline; Group 2 was a positive-control that received orally acrylamide (20 mg/kg/bw) for consecutive 4 weeks; and Group 3 received TPJ; 2 mL/kg/bw, orally for 4 weeks. Rats in Group 4 received pomegranate juice and acrylamide as described in Groups 2 and 3, with the TPJ delivered 1 week before the acrylamide. Acrylamide elevated serum levels of inflammatory cytokines (interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-α)). Moreover, acrylamide decreased concentrations of follicle stimulating hormone (FSH), leutinizing hormone (LH), and testosterone and altered semen characteristics (decreased sperm viability and concentration, and increased sperm abnormalities). Acrylamide increased malondialdehyde levels (MDA), while it decreased antioxidant activities (superoxide dismutase; SOD and reduced glutathione; GSH). The inflammation associated cytokines were restored by TPJ administration to acrylamide rats. GSH and SOD were significantly recovered to near control levels in TPJ plus acrylamide group compared to acrylamide-administered rats. TPJ preadministration to rats restored semen profiles, alteration in testis pathology, and normalized the changes on the male reproductive hormones affected by acrylamide. Furthermore, the TPJ reversed the upregulation in caspases-3 and the decrease in B-cell lymphoma-2 (Bcl-2) gene expressions affected by acrylamide, with significant upregulation of hemoxygenase-1 (HO-1) and nuclear factor erythroid-2 (Nrf2) mRNA expression. These findings collectively revealed that TPJ possesses anti-inflammatory, potent antioxidant, and antiapoptotic effect against acrylamide-induced testicular damage.

The purpose of this study was to investigate the linkage of the association of azoospermia phenotype with genetic alterations, involved in genome instability. Male infertility is a multifactorial pathology, and genetic alterations might be the underlying factors in majority of cases of severe male infertility. The recent emergence of next-generation sequencing offers an opportunity to analyze many genes and their interactions at once, and whole-exome sequencing (WES) together with whole-genome sequencing (WGS) was recently suggested for implementation of diagnosis workup in severe infertility cases. However, the reports on WES in conjunction with burden tests and gene network analysis are scarce or lacking in cases of severe male infertility. WES was performed on 21 nonobstructive azoospermia patients. DNA samples were sequenced using the Twist Comprehensive Exome Panel. Genetic burden test was performed with Testing Rare vAriants using Public Data. Protein interactions were investigated with ConsensusPathDB and Cytoscape. For single nucleotide variants and copy number variations (CNV) analysis, samples were analyzed with the Illumina’s BaseSpace Variant Interpreter. Genetic variant burden was found elevated in 1,473 genes out of 30,000 known testis expressed genes. Three hundred and two genes with increased loss-of-function (LoF) variant set were present in more than one sample. Overrepresentation analysis with pathway-based set of genes with high variant burden demonstrated 26 pathways. Overrepresentation analysis with protein complex-based gene sets obtained 14 sets, showing the involvement in cell proliferation and DNA repair. Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) network analysis with Cytoscape identified two clusters: (1) genes, involved in DNA binding/condensation and repair processes and (2) genes with the role in ribosome biosynthesis and gene expression processes. Increased loss of function germline variant burden and sumoylation may have critical significance in spermatogenesis. These parameters may be used for focused diagnosis in nonobstructive azoospermia patients. This may have both general significance for the decreased organism functionality but in particular is critical in spermatogenesis.