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The aim of this study is to evaluate the beneficial effects of N-acetyl-cysteine (NAC), alpha lipoic acid (ALA) and combination of NAC + ALA supplement in freezing medium on Sperm structural and functional in asthenoteratozoospermia patients. Thirty freshly ejaculated semen samples were cryopreserved with sperm freezing medium (SFM) as control group and three group that SFM supplemented with NAC, ALA and their combination NAC+ ALA. The sperm samples were analysed according to WHO. Mitochondrial membrane potential (MMP), acrosome reaction (AR), antioxidant enzymes and DNA fragmentation were assessed using by Rhodamine123, PSA- FITC ELISA and TUNEL staining respectively. Expression level of NRF2 was assessed by real-time PCR assay. NAC and ALA alone significantly improved sperm motility, viability and DNA fragmentation (p < 0.05). MMP increased in NAC and ALA separately (p < 0.05). While did not affect the amount of sperm morphology and AR (p > 0.05). Antioxidant enzymes significantly difference in NAC and ALA groups (p < 0.05). In addition, NAC and ALA groups showed a significantly higher expression of NRF2 gene compared with other groups (p < 0.05). Our results revealed that the ALA and NAC supplements had a protective effect in cryopreservation process on the structural and functional characteristics of sperm., (© 2022 Institute of Vertebrate Biology CAS. Ecology and Evolution published by John Wiley & Sons Ltd.)

This study aimed to investigate the effects of metformin and forskolin independently and in combinations on the sperm quality parameters and sexual hormones of diabetic male rats. Fifty adult male rats were divided randomly into five identical groups, and diabetes mellitus was induced to the rats, except for the rats in the control group, using a high-fat diet and injection of Streptozotocin. Daily administration of metformin and forskolin independently and in combinations were performed for 8 weeks in different groups. Sperm quality parameters (including sperm count, morphology, sperm motility and Johnson score), testosterone, blood sugar level, Bax to Bcl-2 ratio mRNA expression level and oxidative stress levels were measured and compared between the investigated groups. Treating diabetic rats with metformin and forskolin resulted in significant improvement in sperm quality parameters, increased testosterone levels, reduced oxidative stress in blood and testicular tissue, and decreased blood sugar, and Bax to Bcl-2 ratio level. Although the combination of metformin with forskolin had a higher effect in some parameters such as testosterone levels compared to treatment with metformin or forskolin alone, this combination had not shown a synergistic effect in all the sperm quality parameters. Metformin and forskolin are effective anti-diabetic agents, which significantly improve the sperm quality and sexual hormone levels in diabetic rats. Combining metformin and gorskolin resulted in significantly better testosterone level and antioxidant activity in blood serum without significant effect on sperm quality of diabetic rats., (© 2022 Wiley-VCH GmbH.)

There are a number of risk factors, especially viral diseases, which can lead to infertility. Among the various viral infections, much attention has been given to the role of the Papillomaviridae and Herpesviridae. After collecting 82 semen samples (37 teratospermia, 2 asthenozoospermia, 2 oligoasthenospermia, 1 oligospermia, 6 asthenoteratospermia and 34 normal semen samples), and washing them, the DNA from both freshly ejaculated spermatozoon and washed spermatozoa was extracted. Subsequently, the prevalence of EBV, CMV, HSV-1, HSV-2, VZV and HPV was evaluated using Multiplex PCR and Nested PCR. In this study, 1 normal and 5 abnormal semen samples were infected with HSV-1 (1 normal, 4 teratospermia and 1 oligoasthenospermia). In addition, there were 2 VZV-positive samples (both were teratozoospermia). Nested PCR indicated that 1 asthenozoospermia, 1 asthenoteratospermia, 3 teratospermia and 4 normal samples were HPV positive (including 8 HPV-18 and 1 HPV-33). Among 9 HPV-positive subjects, 3 samples were negative after washing the infected samples. The prevalence of EBV, CMV, VZV, HSV-1 and HSV-2 remained unchanged prior to and after washing. Maybe sperm washing can be useful to eliminate HPV infection from semen samples, but further investigation is required because of the small number of samples., (© 2021 Wiley-VCH GmbH.)

In infertile men, reduced expression of heat shock protein A2 (HSPA2) is related to reduced sperm quality and function. The present study has aimed to investigate the effects of N-acetyl-cysteine (NAC) supplementation on expression of heat shock protein A2 (HSPA2). In this study in continuation of previous study, semen samples from 50 astheno-terato-zoospermic men who have received NAC (600 mg/day) orally for three months were evaluated for expression HSPA2 using RT-PCR, and Western blot analysis. In addition, semen samples of these individuals were assessed for sperm parameters, DNA fragmentation (TUNEL), protamine deficiency (CMA3), lipid peroxidation index (MDA) and total antioxidant capacity (TCA). All assessment was carried out before and after NAC treatment. In addition to improved sperm parameters and aforementioned functional parameters, the presented results revealed the significant increase in relative expression levels of HSPA2 was obtained after using NAC treatment (p < .05). Correlation analysis also demonstrated that HSPA2 expression is significantly related to most of the assessed parameters. NAC may directly or indecently impose its beneficial effect through increased expression of HSPA2, which plays a potential role in proper folding of element needed to counteract stress condition in infertile individuals., (© 2021 Wiley-VCH GmbH.)

The molecular mechanisms of drug use on sexual health are largely unknown. We investigated, the relationship between heroin use disorder and epigenetic factors influencing histone acetylation in sperm cells. The volunteers included twenty-four 20- to 50-year-old men with a normal spermogram who did not consume any drugs and twenty-four age- to BMI-matched men who consume only the drug heroin for more than last four months. HDAC1 and HDAC11 mRNA expression levels in spermatozoa and miR-34c-5p and miR-125b-5p expression levels in seminal plasma were measured. The heroin-user group showed significantly increased white blood cell counts and decreased sperm motility and survival rates (8.61 ± 1.73, 21.50 ± 3.11, 69.90 ± 4.69 respectively) as compared to the control group (1.49 ± 0.32, 38.82 ± 3.05, 87.50 ± 0.99 respectively) (p ≤ .001). An increase in DNA fragmentation index (DFI) (heroin-user group: 41.93 ± 6.59% and control group: 10.14 ± 1.43%, p = .003), a change in frequency of HDAC1 (heroin-user group: 1.69 ± 0.55 and control group: 0.45 ± 0.14, p = .045) and HDAC11 (heroin-user group: 0.29 ± 0.13 and control group: 2.36 ± 0.76, p = .019) in spermatozoa and a significant decrease in seminal miR-125b-5p abundance (heroin-user group: 0.37 ± 0.11 and control group: 1.59 ± 0.47, p = .028) were reported in heroin consumers. Heroin use can lead to male infertility by causing leukocytospermia, asthenozoospermia, DFI elevation in sperm cells and alterations in seminal RNA profile., (© 2020 Wiley-VCH GmbH.)

The objective was to evaluate the effect of inclusion of 2.5% and 5% ovine serum, enriched with vitamin E (Vit E) and fish oil (FO), in human sperm freezing medium. Serum samples were prepared from sixteen rams (n = 4) feeding on a without supplemented diet, and diets supplemented with Vit E, FO and Vit E + FO. Semen samples, from 60 normozoospermic men, were frozen in: (I) a commercial freezing medium (SpermFreeze™; control medium), (II) the commercial freezing medium containing foetal bovine serum, (III) the commercial freezing medium + nonenriched serum (serum group), (IV) the commercial freezing medium + Vit E enriched serum (Vit E group), (V) the commercial freezing medium + FO enriched serum (FO group) and (VI) the commercial freezing medium + Vit E + FO enriched serum (Vit E + FO group). Sperm total and progressive motility, morphology, viability and plasma membrane integrity were significantly higher (p ≤ .05) in Vit E and Vit E + FO groups compared with the control group. Mitochondrial membrane potential did not differ between treatments (p > .05). It was concluded that ovine serum enriched with vitamin E and vitamin E + FO improved the quality of human spermatozoa but enriched serum containing FO could not improve the sperm cryo-injuries., (© 2020 Blackwell Verlag GmbH.)

Considering infection/inflammation to be an important risk factor in male infertility, the aim of this study was to make a comprehensive evaluation of the prevalence of urogenital tract infection/inflammation and its potential impact on sperm retrieval in azoospermic patients. In this prospective study, 71 patients with azoospermia were subjected to an extensive andrological workup including comprehensive microbiological diagnostics (2-glass test, semen, testicular swab and testicular tissue analysis) and testicular biopsy/testicular sperm extraction (TESE). Medical history suggested urogenital tract infection/inflammation in 7% of patients, 11% harboured STIs, 14% showed significant bacteriospermia, 15% had seminal inflammation, 17% fulfilled the MAGI definition, and 27% had relevant pathogens. At the testicular level, 1 patient had a swab positive for bacteria, no viruses were detected, tissue specimens never indicated pathogens, whereas histopathology revealed focal immune cell infiltrates in 23% of samples. Testicular sperm retrieval rate was 100% in obstructive and 46% in nonobstructive azoospermia. None of the infection/inflammation-related variables was associated with the success of sperm retrieval or inflammatory lesions in the testis. The high prevalence of urogenital infection/inflammation among azoospermic men underpins their role as significant aetiologic factors in male infertility. However, this observation does not refer to the chances of sperm retrieval at the time of surgery/TESE., (© 2019 The Authors. Andrologia published by Blackwell Verlag GmbH.)

Chimerism, a rare human disorder, is assumed to be the result of an amalgamation of two separate zygotes in a single embryo. Studies have shown that the phenotypic spectrum of chimerism is variable and there is no definite genotype-phenotype correlation in patients with chimerism, therefore a majority of cases might remain undiagnosed. This study aims to investigate the possible mechanism of the chimerism in a 46,XX/46,XY infertile and phenotypically normal male, with 46,XX blood karyotype and normal spermatogenesis. We have used Interphase-FISH analysis to study the CEPX and CEPY regions on buccal and urine samples as well as molecular analysis of polymorphic short tandem repeats (STR) markers from 34 loci in order to discover the origin of 46,XX/46,XY. Analysis of X-linked and autosomal STR markers on blood, buccal tissue, urine, fibroblast and testis biopsy samples of the proband along with the blood sample of the patient's parents and siblings, showed divergent karyotypes in different tissues and tetragametic chimerism was diagnosed., (© 2019 Blackwell Verlag GmbH.)

In this study, we aimed to determine androgen receptor (AR) and SRD5A2 gene mutations in 45 patients characterised by 46,XY Disorders of Sex Differentiation (DSD) signs with normal testicular development referred to the Children's Medical Center from February 2015 to September 2017. Karyotype and sex hormone analyses were performed. Cytogenetic investigation showed that seven patients were 46,XX DSD, six patients were chromosomal DSD and 32 patients were 46,XY DSD. Eight exons of the AR gene and five exons of the SRD5A2 gene were amplified. Two cases were affected with androgen insensitivity syndrome (AIS) (missense mutation on exon 7, position c.3637 G>A: p.R841H and position c.3610 G>A: p.R832Q), one case was affected with 5-alpha-reductase deficiency type 2 (missense mutation at c.578A>G: p.N193S on exon 4), and 22 cases (88%) did not demonstrate AIS or 5α-RD2 gene abnormality. Due to the great impact of these disorders on human lifestyle, evaluation of genes involved can improve genetic counselling and therapeutic management. We focused on the AR and SRD5A2 genes in patients with 46,XY DSDs with normal testicular development referred to the Children's Medical Center from all over the country to eventually culminate in a reliable prenatal diagnosis protocol at this major referral centre giving service to a great number of families with consanguineous marriages., (© 2019 Blackwell Verlag GmbH.)

The present study was designed to assess the responsiveness of hypothalamic-pituitary-gonadal axis to kisspeptin administration with increasing age in men. Human kisspeptin-10 was administered in single iv bolus dose (1 µg/kg BW) to healthy adult, middle and advanced age men. Serial blood samples were collected for 30 min pre- and 120 min post-kisspeptin injection periods at 30-min interval. Analysis of plasma LH by ELISA showed a significant (p < 0.05) increase after kisspeptin-10 administration in all groups, whereas plasma testosterone concentration was significantly elevated (p < 0.05) after kisspeptin-10 injection only in the adult men group. Present results suggest that in men, central hypothalamic-pituitary axis remains active and shows responsiveness to kisspeptin stimulation across life. However, Leydig cell responsiveness to kisspeptin-induced LH decreases with age in men., (© 2018 Blackwell Verlag GmbH.)

The present study was performed to show the ameliorative effect of berberine (BBR), as an antioxidant and anti-inflammatory agent, against experimental varicocele (VCL)-induced molecular and histological damages. For this purpose, 50 mature Wistar rats were divided into control, control-sham, VCL-sole, 50 mg/kg and 100 mg/kg BBR-treated VCL-induced groups. The tissue levels of interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), nitric oxide (NO), total antioxidant capacity (TAC), malondialdehyde (MDA), superoxide dismutase (SOD) and gluthatione peroxidase (GSH-px) as well as the mRNA levels of testicular CuZn SOD, MnSOD, EC-SOD and GSH-px were evaluated. The serum concentration of testosterone and germ cells mRNA damage were analysed. Finally, the sperm viability, motility, DNA integrity and chromatin condensation were analysed. Observations revealed that, the BBR significantly downregulated VCL-increased IL-6, TNF-α and NO levels, upregulated the CuZn SOD, MnSOD, EC-SOD and GSH-px mRNA level, decreased testicular MDA content, enhanced serum testosterone level and ameliorated testicular TAC, SOD and GSH-px levels. The animals in BBR-treated groups exhibited diminished mRNA damage versus non-treated VCL-induced group. The BBR has significantly (p < 0.05) improved sperm parameters. In conclusion, the BBR by promoting testicular antioxidant potential and by downregulating inflammatory reactions fairly promotes spermatogenesis and upregulates the sperm quality., (© 2018 Blackwell Verlag GmbH.)

Despite scientific advances, many of the treatments in male infertility remained indeterminate. In recent years, the attention to herbal remedies as an effective treatment for male infertility is considerable. We designed this study to determine the effects of Alpinia officinarum on the results of semen analysis in men with idiopathic infertility. In this clinical trial, seventy-six participants with idiopathic infertility were included in the intervention (plant treatment: n = 31; placebo: n = 29). Participants were randomised to take capsules containing dried extract of A. officinarum rhizome or placebo on a daily (total daily dosage of 300 mg) basis for 3 months. After 12 weeks of intervention, the sperm count and total number of spermatozoa with normal morphology were increased in participants treated with A. officinarum extract compared with the placebo group. The mean sperm count was initially 52 × 10 6  ± 24 × 10 6 /ml which changed to 71 × 10 6  ± 23 × 10 6 /ml, after intervention (p = 0.043). Also, the mean percentage of spermatozoa with normal morphology was 14.34% ± 9.16% before the treatment which significantly increased to 19% ± 14.89% (p < 0.001). Alpinia officinarum, a traditional medicine remedy, can be effective in the improvement of sperm morphology and sperm count in idiopathic infertility without causing adverse effects., (© 2018 Blackwell Verlag GmbH.)

Androgen receptor (AR) mediates androgen activities such as the growth of accessory sex organs, and initiation and promotion of spermatogenesis. There are two trinucleotide polymorphisms (CAG and GGN repeats) in the first exon of AR gene that their association with infertility is still controversial. The variants of both polymorphic repeats were investigated by PCR-Sequencing in 220 infertile men (80 azoospermic, 60 oligospermic and 80 asthenospermic) and 80 healthy fertile controls. AR Expression level was quantified by RT-qPCR on 30 patients (20 patients with nonobstructive azoospermia (NOA) and 10 obstructive azoospermia patients as controls). Our results demonstrated that the medians of CAG and GGN repeats length in infertile group were significantly higher than fertile men (p < 0.05). AR expression results showed a significant increase in SCOS group compared to control (p < 0.05). Long stretches of tandem repeats of AR gene may negatively affect the function of the gene and consequently lead to male infertility. In patients with SCOS, AR expression increases because of the lack of germ cells. Therefore, with increasing AR expression, the probability of SCOS occurrence is also increased. It can be concluded that increasing AR expression in testes tissue decreases the probability of sperm presence., (© 2018 Blackwell Verlag GmbH.)

Diabetes causes a systemic inflammatory response with increased oxidative stress and can adversely effect spermatogenesis. Sickle-weed plant, scientifically called Falcaria vlugaris, contains antioxidants and antimicrobial ingredients. The aim of this study was to determine the effect of Falcaria vulgaris vulgaris extract on the sperm parameters of diabetic rats. In this experimental study, diabetes was induced in the animals using streptozotocin (STZ; 60 mg/kg). Sixty-four rats were equally divided into eight groups as follows: nondiabetic (normal) controls; normal rats given 50, 100 or 150 mg/kg doses of F. vulgaris extract; diabetic controls induced by STZ; and diabetic animals that received STZ plus 50, 100 or 150 mg/kg F. vulgaris extract. The animals were injected with the extract intraperitoneally once a day for 28 consecutive days. The sperm parameters of count, motility, morphology, viability, total antioxidant capacity and seminiferous tube diameter were analysed and compared. The results indicated that the values of all parameters decreased significantly in the diabetic control groups compared to the normal control group (p < 0.00). The parameters of the diabetic groups that received 50, 100 or 150 mg/kg F. vulgaris extract showed significant increases in all parameters at all doses compared to the diabetic control group (p ≤ 0.001). The administration of F. vulgaris extract as a potent antioxidant was able to compensate for the toxicity induced by STZ and increase the quality of some spermatozoa., (© 2018 Blackwell Verlag GmbH.)