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  • Effects of taking tadalafil 5 mg once daily on erectile function and total testosterone levels in patients with metabolic syndrome
    We aimed to evaluate the efficacy of tadalafil 5 mg once-daily treatment on testosterone levels in patients with erectile dysfunction (ED) accompanied by the metabolic syndrome. A total of 40 men with metabolic syndrome were evaluated for ED in this study. All the patients received 5 mg tadalafil once a day for 3 months. Erectile function was assessed using the five-item version of the International Index of Erectile Function (IIEF) questionnaire. Serum testosterone, follicle-stimulating hormone and luteinising hormone levels were also evaluated, and blood samples were taken between 08.00 and 10.00 in the fasting state. All participants have three or more criteria of metabolic syndrome. At the end of 3 months, mean testosterone values and IIEF scores showed an improvement from baseline values (from 3.6 ± 0.5 to 5.2 ± 0.3, from 11.3 ± 1.9 to 19 ± 0.8 respectively). After the treatment, serum LH levels were decreased (from 5.6 ± 0.6 to 4.6 ± 0.5). There was significantly difference in terms of baseline testosterone and luteinising hormone values and IIEF scores (p < .05). Based on our findings, we recommend tadalafil 5 mg once daily in those men with erectile dysfunction especially low testosterone levels accompanied by metabolic syndrome.
  • Cumene hydroperoxide induced changes in oxidation–reduction potential in fresh and frozen seminal ejaculates
    Oxidation–reduction potential (ORP) is a newer integrated measure of the balance between total oxidants (reactive oxygen species—ROS) and reductants (antioxidants) that reflects oxidative stress in a biological system. This study measures ORP and evaluates the effect of exogenous induction of oxidative stress by cumene hydroperoxide (CH) on ORP in fresh and frozen semen using the MiOXSYS Analyzer. Semen samples from healthy donors (n = 20) were collected and evaluated for sperm parameters. All samples were then flash-frozen at −80°C. Oxidative stress was induced by CH (5 and 50 μmoles/ml). Static ORP (sORP—(mV/106 sperm/ml) and capacity ORP (cORP—μC/106 sperm/ml) were measured in all samples before and after freezing. All values are reported as mean ± SEM. Both 5 and 50 μmoles/ml of CH resulted in a significant decline in per cent motility compared to control in pre-freeze semen samples. The increase in both pre-freeze and post-thaw semen samples for sORP was higher in the controls than with 50 μmoles/ml of CH. The change from pre-freeze to post-thaw cORP was comparable. The system is a simple, sensitive and portable tool to measure the seminal ORP and its dynamic impact on sperm parameters in both fresh and frozen semen specimens.
  • Fertility enhancing activity and toxicity profile of aqueous extract of Chasmanthera dependens roots in male rats
    This study evaluated the fertility-enhancing activity and safety of aqueous extract of Chasmanthera dependens root (AECDR) in male rats. In the fertility study, twenty, sodium arsenite (10 mg/kg) body weight (BW)-treated male rats (171.02 ± 3.36 g), assigned into four groups (I–IV), received 1 ml of distilled water (DW), 25, 50 and 100 mg/kg BW of AECDR for 60 days, whereas the control received DW. After 7 days of pairing with female rats (153.67 ± 2.24 g), spermatogenic, fertility, testicular function indices and enzymatic antioxidant activities were evaluated. The animal groupings in the toxicity study were similar to the fertility study except no administration of sodium arsenite. Sodium arsenite treatment-related decreases (p < .05) in the semen and sperm parameters, testicular function indices, antioxidant activities and female rat fertility indices were reversed/ameliorated by AECDR. AECDR significantly altered the function indices of the liver and kidney and the lipid profile and selectively altered the haematological parameters. There was no treatment-related histoarchitectural changes in the organs. Overall, the aqueous extract of C. dependens roots exhibited pro-spermatogenic, fertility enhancing, antioxidant and androgenic activities in male rats. It also exhibited functional toxicity. Therefore, the chronic use of AECDR may not be completely safe as oral remedy.
  • Sperm DNA fragmentation as a result of ultra-endurance exercise training in male athletes
    Intensive sports practice seems to exert negative effects on semen parameters; in order to assess these effects, the objective of this study was to assess semen, including DNA fragmentation, and hormone parameters in elite triathletes. Twelve high-level triathletes preparing for a National Triathlon Championship participated in the study. The qualitative sperm parameters analysed were volume, sperm count, motility, morphology and DNA fragmentation; when needed, additional testing was performed. Assessed hormones were testosterone (T), cortisol (C) and testosterone–cortisol ratio (T/C). Maximum oxygen consumption and training characteristics were also assessed. Hormonal values and physical semen parameters were within normal ranges. DNA fragmentation showed high values (20.4 ± 6.1%). Round cells in semen were higher than normal (2.8 ± 1.5 million/ml), with the presence of macrophages. Correlations were found for several parameters: concentration of round cells positively correlated with progressive sperm motility (p = .01) and sperm morphology (p = .02); contrarily, the correlation found with DNA fragmentation was negative (p = .04). Sperm DNA fragmentation and the T/C ratio, however, were correlated in a positive manner (p = .03). As evidenced by the observed results, sperm DNA fragmentation is affected by high-level sports practice; therefore, high loads of endurance training could potentially interfere with the athlete's fertility potential.
  • Icariin improves the sexual function of male mice through the PI3K/AKT/eNOS/NO signalling pathway
    We aimed to investigate the effect and mechanism of icariin on male sexual function. Forty-eight Crl:CD1(ICR) male mice were randomly divided into control, low-, medium- and high-dose icariin group (intragastric administration of 50, 100 and 200 mg/kg/d for 21 days). Mating experiment was then performed at a ratio of 1: 3 (male: female). The mating behaviours of male mice were recorded. The genital indexes and serum testosterone, nitric oxide (NO), hypothalamic dopamine (DA) and 5- hydroxy tryptophan (5-HT) concentrations were measured. The expression of endothelial nitric oxide (eNOS), phosphatidylinositol tallow alcohol 3-kinase (PI3K) and phosphorylated protein kinase (p-AKT) in penile tissue was detected by Western blot. All icariin groups exhibited shorter capture latency and ejaculation latency, increased number of capture and ejaculation, higher capture and ejaculation rate, and higher testicular and prostate indexes compared with controls (p < .001). These groups had higher serum testosterone and NO concentrations (p < .001), hypothalamic DA and 5-HT levels, and eNOS, PI3K and phosphorylated AKT expressions in penile tissue (p < .05). The effect of icariin was dose-dependently increased. Our study suggests that icariin improves the sexual function of male mice, which might be associated with the hypothalamic–pituitary–gonadal axis and the PI3K/AKT/eNOS/NO signalling pathway.
  • Sperm DNA methylation of H19 imprinted gene and male infertility
    Infertility affects up to 15% of reproductive-aged couples worldwide, with male factor being detected in 40%–50% of the cases. Proper sperm production is associated with the establishment of appropriate epigenetic marks in developing germ cells. Several studies have demonstrated the association between abnormal spermatogenesis and epigenetic disturbances with the major focus on DNA methylation. Imprinted genes are expressed in a parent-of-origin-specific manner, and the role of their DNA methylation in proper spermatogenesis has been documented recently. The existing evidence along with the absence of relevant data in south of Iran prompted us to study the methylation of H19 imprinted gene in spermatozoa of idiopathic infertile patients (males with abnormalities in sperm parameters) and healthy controls by Combined Bisulfite Restriction Analysis. According to our results, the lowest methylation percentage of H19 imprinted gene belongs to three cases with sperm characteristics under normal range (two cases Oligoasthenoteratozoospermia and one case Oligoteratozoospermia). However, our results show that the median of methylation percentage for H19 is not statistically significant between case and control groups. Our results and those of others introduce DNA methylation as a potential marker of fertility and should be investigated with more patients in future studies.
  • Effects of various cryoprotectants on the quality of frozen–thawed immature bovine (Qinchuan cattle) calf testicular tissue
    To investigate the effects of different concentrations of various cryoprotectants (CPs) on the cell viability as well as expression of spermatogenesis-related genes, such as CREM, Stra8 and HSP70-2 in frozen–thawed bovine calf testicular tissue, immature bovine (Qinchuan cattle) calf testicular tissue was collected and cryopreserved in the cryomedia containing different concentrations (5%, 10%, 15% and 20%) of the following three CPs: glycerol, ethylene glycol (EG) and dimethyl sulphoxide (DMSO) respectively. After 1 month cryopreservation in liquid nitrogen, cell viability was evaluated using Trypan blue exclusion under a bright-field microscope. The mRNA expression of the three genes was also evaluated using qRT-PCR. The results indicated that different concentrations of glycerol, EG and DMSO in cryomedia during cryopreservation could protect bovine calf testicular tissue in various ways to avoid freezing or cryopreservation-induced expression changes in spermatogenesis-related genes. The highest cell viability and the three spermatogenesis-related genes (CREM, Stra8 and HSP70-2) expression level came from the cryomedia containing glycerol, EG and DMSO at 10% concentration respectively (p < .05). Meanwhile, compared with the other CPs, the frozen–thawed bovine calf testicular tissue treated with 10% DMSO exhibited the highest cell viability and mRNA expression level of the spermatogenesis-related genes (CREM, Stra8 and HSP70-2).
  • Androgen deficiency impairs erectile function in rats through promotion of corporal fibrosis
    The aim of this study was to investigate the underlying mechanism of androgen deficiency inducing corporal fibrosis, thereby causing erectile dysfunction (ED). Forty 12-week-old healthy male rats were divided randomly into four groups: normal control group (Control); castration group (Castration); the other 20 rats were castrated followed by testosterone (T) (orally) each day: castration + 10mg/kg T group (Castration + 10T) and castration + 20 mg/kg T group (Castration + 20T). After 8 weeks' treatment, the main outcome measures were the following: serum levels of T; the ratios of intracavernous pressure (ICP) to mean arterial pressure (MAP); histologic changes in penile smooth muscle cells; the Smad and non-Smad pathways; and extracellular matrix (ECM) protein deposition. Castration group showed lower level of T and ratio of ICP/MAP, reduced ratio of penile smooth muscle cells/collagen, increased extracellular matrix protein deposition, and a higher expression of the Smad and non-Smad pathways. Castration + 10T partially preserved erectile function and histology stabilisation. However, the Castration + 20T group showed significantly better erectile function and molecular changes. Better efficacy could be expected with ART of adequate dose. Androgen deficiency induces corporal fibrosis through activation of the Smad and non-Smad pathways, and accumulation of ECM proteins.
  • Single nucleotide polymorphisms associated with nonsyndromic cryptorchidism in Mexican patients
    Cryptorchidism is a frequent genitourinary malformation considered as an important risk factor for infertility and testicular malignancy. The aetiology of cryptorchidism is multifactorial in which certain SNPs, capable of inhibiting the development of the gubernaculum, are implicated. We analysed 16 SNPs by allelic discrimination and automated sequencing in 85 patients and 99 healthy people, with the objective to identify the association between these variants and isolated cryptorchidism. In two different patients with unilateral cryptorchidism, we found the variants rs121912556 and p.R105R of INSL3 gene in a heterozygous form associated with cryptorchidism, so we could considered them as risk factors for cryptorchidism. On the other hand, SNPs rs10421916 of INSL3 gene, as well as the variants rs1555633 and rs7325513 in the RXFP2 gene, and rs3779456 variant of the HOXA10 gene were statistically significant, when the patients and controls were compared and could be considered as protective factors since are predominantly present in controls. The genotype–phenotype correlation did not show statistical significance. With these results, we could conclude that these polymorphisms can be considered as important variants in our population and would contribute in the future knowledge of the aetiology and physiopathology of cryptorchidism.
  • Significant predictive factors for subfertility in patients with subclinical varicocele
    We made a retrospective study to determine useful parameters for predicting subfertility in patients with subclinical varicocele (SV). One hundred and fifty men with SV and 17 age-matched men without SV were divided into three groups: Group 1, subfertile patients (n = 15); Group 2, fertile patients (n = 135); and Group 3, control patients (n = 17). Their age, body mass index (BMI), semen analysis, scrotal temperature, testicular volume, resistive index (RI), pulsatility index (PI) and peak retrograde flow (PRF) were compared. Subfertile patients (Group 1) with SV had significantly lower testicular volume and higher scrotal temperature, RI, PI and PRF than fertile men with SV (Group 2) and the control group (Group 3). Elderly men (>50 years, n = 30) with SV had a significantly higher incidence of bilateral SV than young men with SV (10/30; 33.3% vs. 12/120; 10%). There was no difference in age and BMI among the three groups. Patients with SV and RI >0.55 ml/s, PI >0.99 ml/s, total testicular volume <27 cc, scrotal temperature >34.9°C and PRF >29 cm/s have higher incidence of subfertility. Patients with SV may suffer from subfertility regardless of age. Close follow-up with colour Doppler ultrasound may be beneficial.
  • Ferulic acid prevents lead-induced testicular oxidative stress and suppressed spermatogenesis in rats
    Lead affects multiple organ systems including testis. We investigated the effects of ferulic acid (FA) on lead-induced oxidative stress and spermatogenesis suppression in rats. Animals received lead acetate (500 mg/L in drinking water) and/or FA (50 mg/kg, i.g.) for eight weeks. Lead increased testicular malondialdehyde (MDA) and nitrite levels and decreased glutathione (GSH) content and catalase (CAT) activity. Lead decreased testis weight and testosterone level. Sperm parameters decreased in lead group. FA ameliorated the decreased testis weight, serum testosterone as well as sperm count, viability, motility and normal morphology in lead group. FA improved antioxidant capacity as well as sperm count, viability, motility and normal morphology. FA decreased Johnsen's mean testicular biopsy score (MTBS) criteria by restoring degeneration, atrophy and tubular disarrangement. FA also normalised spermatogonia, spermatocytes and spermatids numbers in lead group and led to increases in number of Leydig and Sertoli cells. FA showed beneficial effects in lead-induced testicular oxidative stress and spermatological disorders, through inhibiting lipid peroxidation and enhancing antioxidant defence systems. The positive effects of FA on Leydig cells may be involved in restoring testosterone levels in lead group. FA can be considered a potential candidate to protect testis against the deleterious effect of lead intoxication.
  • Testosterone therapy has positive effects on anthropometric measures, metabolic syndrome components (obesity, lipid profile, Diabetes Mellitus control), blood indices, liver enzymes, and prostate health indicators in elderly hypogonadal men
    To alleviate late-onset hypogonadism, testosterone treatment is offered to suitable patients. Although testosterone treatment is commonly given to late-onset hypogonadism patients, there remains uncertainty about the metabolic effects during follow-ups. We assessed the associations between testosterone treatment and wide range of characteristics that included hormonal, anthropometric, biochemical features. Patients received intramuscular 1,000 mg testosterone undecanoate for 1 year. Patient anthropometric measurements were undertaken at baseline and at each visit, and blood samples were drawn at each visit, prior to the next testosterone undecanoate. Eighty-eight patients (51.1 ± 13.0 years) completed the follow-up period. Testosterone treatment was associated with significant increase in serum testosterone levels and significant stepladder decrease in body mass index, total cholesterol, triglycerides and glycated haemoglobin from baseline values among all patients. There was no significant increase in liver enzymes. There was an increase in haemoglobin and haematocrit, as well as in prostate-specific antigen and prostate volume, but no prostate biopsy intervention was needed for study patients during 1-year testosterone treatment follow-up. Testosterone treatment with long-acting testosterone undecanoate improved the constituents of metabolic syndrome and improved glycated haemoglobin in a stepladder fashion, with no adverse effects.
  • Improvement in erectile function in a rat model of high cholesterol diet-induced atherosclerosis by atorvastatin in a manner that is independent of its lipid-lowering property
    The purpose of the present study is to explore the effects of a lipid-lowering drug atorvastatin, a three-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, in the treatment of erectile dysfunction (ED) in a rat model of atherosclerosis (AS) and the possible mechanisms underneath. A high-cholesterol diet was administrated to Sprague-Dawley rats in an attempt to induce an ASED model, which was later confirmed by abdominal aorta histopathology and erectile function evaluation. ASED rats were further assigned to non-treatment group, atorvastatin low-dose treatment group (5 mg kg−1 day−1), high-dose group (10 mg kg−1 day−1) and sildenafil (1.5 mg kg−1 day−1) treatment group. Lipid profile, erectile function, oxidative stress biochemical markers, endothelial nitric oxide synthase (eNOS) and extracellular superoxide dismutase (SODEX) mRNA expression were evaluated after 8-week treatment duration. Erectile function was impaired in AS rat model, which was preserved in atorvastatin and sildenafil intervention groups. The oxidative stress biochemical markers were attenuated, while eNOS and SODEX mRNA expression were restored in atorvastatin and sildenafil groups, which were found to be involved in ED pathogenesis. However, the lipid profile remained unaltered in the treatment group, and it was elevated in ASED rats. This kind of lipid-lowering agent, or atorvastatin, has the utilisation potential in ASED treatment, even before lipid profiles altered. This effect on erectile function preservation of atorvastatin was attributed to its preservation of endothelial function, possibly through amelioration of oxidative stress and improvement in eNOS expression.
  • Efficacy and safety of testosterone replacement gel for treating hypogonadism in men: Phase III open-label studies
    Efficacy and safety of testosterone gel 2% (TG) were evaluated in two phase 3, open-labelled, single-arm, multicentre studies (000023 and extension study 000077). Hypogonadal men having serum testosterone levels <300 ng/dl at two consecutive measurements were included. Study duration was 9 months (000023: 3 months; 000077: 6 months). Starting dose of TG (46 mg) was applied on upper arm/shoulder. The primary endpoint (000023) was responder rate (subjects with average 24-hour serum testosterone concentration 300-1050 ng/dl on Day 90). Study 000077 evaluated the safety of TG in patients rolling over from study 000023 over a period of 6 months. Of 180 subjects in 000023, 172 completed and 145 rolled over to 000077, with 127 completers. The responder rate was 85.5%. Fewer subjects in 000077 (12.7%) versus 000023 (31.8%) had maximum testosterone concentration (Cmax) >1500 ng/dl, with no significant safety concerns. Significant improvements in sexual function and quality of life were noted in both studies. Subjects experienced few skin reactions without notable increases in prostate-specific antigen and haematocrit levels. TG was efficacious with an acceptable safety profile. Cmax >1500 ng/dl did not exhibit distinct impact on safety parameters. However, further optimisation of titration schema to reduce Cmax is warranted while maintaining the average steady state total testosterone concentration.
  • Long-term feeding of hydroalcoholic extract powder of Lepidium meyenii (maca) enhances the steroidogenic ability of Leydig cells to alleviate its decline with ageing in male rats
    This study examined whether feeding hydroalcoholic extract of Lepidium meyenii (maca) to 8-week-old (sexually maturing) or 18-week-old (mature) male rats for more than a half year affects serum testosterone concentration and testosterone production by Leydig cells cultured with hCG, 22R-hydroxycholesterol or pregnenolone. Testosterone concentration was determined in the serum samples obtained before and 6, 12, 18 and 24 weeks after the feeding, and it was significantly increased only at the 6 weeks in the group fed with the maca extract to maturing rats when it was compared with controls. Testosterone production by Leydig cells significantly increased when cultured with hCG by feeding the maca extract to maturing rats for 27 weeks (35 weeks of age) and when cultured with 22R-hydroxycholesterol by feeding it to mature rats for 30 weeks (48 weeks of age). Overall testosterone production by cultured Leydig cells decreased to about a half from 35 to 48 weeks of age. These results suggest that feeding the maca extract for a long time to male rats may enhance the steroidogenic ability of Leydig cells to alleviate its decline with ageing, whereas it may cause only a transient increase in blood testosterone concentration in sexually maturing male rats.
  • Effect of Kaempferia parviflora on sexual performance in streptozotocin-induced diabetic male rats
    Kaempferia parviflora Wall. Ex.Baker or Krachidum (KP) has been used locally in medicine and food. It has been claimed that KP has aphrodisia properties; however, no scientific data in support of this function in diabetic model have been reported. This study aimed to investigate the efficacy of KP on sexual behaviour and sperm parameter in streptozotocin (STZ)-induced diabetic male rats. Diabetes was induced in twenty male rats by STZ and divided into four groups: diabetic control group, and 3 treatment groups where KP was dose at 140, 280 and 420 mg/kg orally once a day for 6 weeks. Five normal control rats were treated with vehicle. The body weight, blood glucose, food intake, epididymal sperm parameter, sexual behaviour and serum testosterone level were evaluated. The results showed that KP treatment has no effect on the body weight, blood glucose and food intake in diabetic rats. A significant increase in sperm density in diabetic rats was observed (p < .05) at highest dose of KP. Furthermore, KP treatment demonstrated a significant recovery of sexual behaviour and serum testosterone levels in diabetic rats. These results confirm that KP exhibits aphrodisiac properties that improve the sperm density, testosterone level and sexual performance of STZ-induced diabetic rats.
  • Mean platelet volume, platelet distribution width and platelet count in erectile dysfunction: A systematic review and meta-analysis
    The aim of this study was to investigate the relationship between mean platelet volume (MPV), platelet distribution width (PDW), platelet count (PC) and erectile dysfunction (ED). We searched for observational studies from PubMed, EMBASE, Web of Science and CNKI up to 31 March 2016. Two reviewers independently selected the studies and extracted the data. MPV, PDW, and PC and mean differences in these platelet indices between healthy subjects and ED patients were explored using the Comprehensive Meta-Analysis software package. Seven studies including 795 patients and 524 healthy subjects met the inclusion criteria. The MPV was significantly larger in patients with ED than controls with the standardised mean difference of 0.596 fL (95% CI: 0.378, 0.815, p < 0.001). In ED patients, the pooled mean difference in MPV between vasculogenic ED patients and nonvasculogenic ED patients was 0.706 fL in case–control studies (95% CI: 0.410, 1.002, p < 0.001). There was no significant difference in PDW and PC between healthy subjects and ED patients. The available data suggest that larger MPV was associated with ED. Patients with vasculogenic ED tend to have higher MPV than nonvasculogenic ED patients. Further studies are needed to assess whether increased MPV in ED patients is associated with increased cardiovascular disease.
  • Testosterone in human studies: Modest associations between plasma and salivary measurements
    Testosterone is involved in many processes like aggression and mood disorders. As it may easily diffuse from blood into saliva, salivary testosterone is thought to reflect plasma free testosterone level. If so, it would provide a welcome noninvasive and less stressful alternative to blood sampling. Past research did not reveal consensus regarding the strength of the association, but sample sizes were small. This study aimed to analyse the association in a large cohort. In total, 2,048 participants (age range 18–65 years; 696 males and 1,352 females) were included and saliva (using cotton Salivettes) and plasma were collected for testosterone measurements. Levels were determined by enzyme-linked immunosorbent assay and radioimmunoassay respectively. Free testosterone was calculated by the Vermeulen algorithm. Associations were determined using linear regression analyses. Plasma total and free testosterone showed a significant association with salivary testosterone in men (adjusted β = .09, p = .01; and β = .15, p < .001, respectively) and in women (adjusted β = .08, p = .004; and crude β = .09, p = .002 respectively). The modest associations indicate that there are many influencing factors of both technical and biological origin.
  • Transformation, migration and outcome of residual bodies in the seminiferous tubules of the rat testis
    Experiments were performed to study the transformation, migration and outcome of residual bodies (RBs) in the seminiferous tubules of the rat testes. One part of the testes from adult Sprague–Dawley rats was used to generate paraffin sections to observe RBs and RB precursors through specific staining, and the other part of the testes was used to generate ultrathin sections to observe RBs under a transmission electron microscope. Deep blue particles of different sizes were observed in some seminiferous tubules through specific staining for RBs and RB precursors. These particles first appeared in the seminiferous tubules at stage I of the spermatogenic cycle, and after spermiation, the particles travelled rapidly towards the deeper region of the seminiferous epithelium and soon appeared close to the basement membrane of the seminiferous tubule. All of the particles in the tubules disappeared at stage IX. Using transmission electron microscopy, components of different electron densities were observed in the RBs on the surface of the seminiferous epithelium, all of which gradually formed in the cytoplasm of spermatozoon in later stages of spermiogenesis. After the spermatozoa were released, the RBs in the epithelium travelled quickly to the edge of the tube and were gradually transformed into lipid inclusions. These lipid inclusions ultimately became lipidlike particles. The lipidlike particles were discharged into the interstitial tissue. RBs initiate their own digestive process before their formation during spermiation in the rat testes. After spermiation, the RBs transform into lipid inclusions and finally into lipidlike particles. These lipidlike particles can be eliminated from the seminiferous tubules.
  • A literature review of antithrombotic and anticoagulating agents on sexual function
    Although millions of people receive antithrombotic agents (ATAs) or anticoagulating agents (ACAs) for vascular prophylaxis daily, the negative impact of these agents on sexual function has not been systematically studied. Therefore, a literature search was conducted to determine the effects of the marketed ATAs and ACAs on sexual function. In regard to men, the results show that thienopyridine derivatives increase the risk of erectile dysfunction (ED) and decrease libido and sexual function. The relationship between aspirin use and ED is inconsistent, ranging from a moderate risk to beneficial effects. Nonetheless, aspirin appears to result in a lower risk for ED than does clopidogrel, and seems to benefit patients with lithium-induced ED. Coumarin can cause vasculogenic priapism. In regard to women, only a single report of genital haemorrhage was found. Available data exclusively focus on male subjects. Taken together, ATAs and ACAs can disturb sexual function in different aspects in men. Newer thienopyridine derivatives, such as prasugrel or ticagrelor, may be used as a substitute for clopidogrel when sexual dysfunction occurs. Priapism and genital haemorrhage were found to be uncommon but serious complications of ACA treatment. Additional studies examining the effects of ATAs and ACAs on sexual function are needed, especially in woman and elderly.
  • Clinical and molecular characteristics in 15 patients with androgen receptor gene mutations from South China
    A variety of mutations in the androgen receptor (AR) gene are linked to androgen insensitivity syndrome (AIS) or sexual development disorder. Here, we studied 15 patients with various degrees of disorders of genital hypoplasia from South China. Clinical data including basal hormone level, phenotype, karyotyping and SRY gene identification were documented. Exons with flanking intronic region of the AR gene were sequenced and analysed for mutations, and a total of eight mutations were identified in the AR gene. Of eight mutations, two novel mutations c.2518G>T (p.Asp840Tyr) and c.1186G>C (p.Gly396Arg) were predicted to be damaging by SIFT and Polyphen2 online software. Previously reported mutations: c.528C>A (p.Ser176Arg), c.1789G>A (p.Ala597Thr), c.2612C>T (p.Ala871Val), c.1752C>A (p.Phe584Leu), c.171_172insCTG (p.57_58insLeu) and c.2659A>G (p.Met887Val) were also detected in our subjects. Most of them are involved in hypospadias, penis dysplasia or other disorders of sexual development. A complete AIS case (p.Phe584Leu) with female phenotype and high serum concentrations of dihydrotestosterone (DHT) was also found. This study presented a wide range of spectrum of AIS (from partial AIS to complete AIS) caused by AR mutations in South China population. It suggests that further study with larger data set need to be performed to elucidate the differences of the phenotypes in our study.
  • Oxidative stress and motility impairment in the semen of fertile males
    The aim of the study was to determine the total oxidant status (TOS) and evaluate the influence of oxidative stress on sperm quality in fertile males. The study population consisted of 55 fertile males. Based on the seminal plasma TOS value, the study subjects were divided into the two subgroups: a group with a low (TOS-L) and a high (TOS-H) value. Comparing the TOS-H group with the TOS-L group, we found poorer sperm motility in the TOS-H group. We found lower total antioxidant capacity values and lower activity levels in the majority of the determined superoxide dismutase, glutathione peroxidase, glutathione-S-transferase and glucose-6-phosphate dehydrogenase. Further, we found higher levels of copper and iron as well as lower levels of zinc in the TOS-H group. We observed lower medians of IL-2, 4, 6, 8 and INF-γ in the TOS-H group compared with the TOS-L group, whereas the medians of IL-1β, IL-10 and IL-12 were significantly higher. In fertile males, higher oxidative stress intensity was associated with poorer semen quality and decreased antioxidant capacity in semen. These negative effects might be a result of decreased activities of antioxidant enzymes and altered levels of trace metals and cytokines.
  • Genetic analysis of HOXA11 gene in Chinese patients with cryptorchidism
    Cryptorchidism is the most common congenital anomaly in male children. Its aetiology remains unknown in the majority of cases. Because HOXA11 plays a vital role in regulating testicular descent, genetic variants in HOXA11 genes may contribute to the risk of cryptorchidism. In this study, mutation analysis was performed on the HOXA11 gene in a cohort of 89 patients with cryptorchidism. Furthermore, an association analysis of the HOXA11 tag single nucleotide polymorphism rs6461992 was performed in 168 patients with unilateral cryptorchidism and 193 controls. No pathogenic mutations were found. A significant difference in genotype and allele distribution was detected between cases and controls (p = .029 and .022 respectively). These results suggest that mutations in the coding sequence of HOXA11 might not be a common cause of cryptorchidism, while common polymorphisms in the HOXA11 gene might contribute to the risk of developing unilateral cryptorchidism.
  • Stenotrophomonas maltophilia isolated from prostatic fluid as an infertility factor in a male dog
    The aim of this case was to describe very rare infection of canine prostate gland with Stenotrophomonas maltophilia which had influence on male fertility. The bacterium was cultured from third fraction of the ejaculate collected by manual manipulation. The sperm concentration and motility parameters were evaluated by Hamilton-Thorne Sperm Analyser, version IVOS 12.3, sperm morphology by Diff-Quick staining and live/dead spermatozoa by eosin/nigrosin staining. After 3 weeks of treatment with targeted antibiotic, ciprofloxacin, there was no bacterial growth in prostate fluid. Semen parameters were improved after 60 days from the end of treatment, and females were successfully mated.
  • Impact of physical activity on patient self-reported outcomes of lifelong premature ejaculation patients: Results of a prospective, randomised, sham-controlled trial
    Previous studies have investigated whether physical activity increases serotonin hormone levels. Serotonin receptor dysfunction is one of the frequently accused factors of premature ejaculation (PE). Nevertheless, no studies up to date have demonstrated that the association between physical activity and premature ejaculation. We aimed to investigate the relationship between physical activity and PE and determine whether moderate physical activity might delay ejaculation time or be an alternative treatment for PE. A total of 105 patients diagnosed with PE were enrolled in this study. Of the patients, 35 were treated with dapoxetine, (30 mg) on demand (Group 1), 35 performed moderate physical activities (Group 2), and 35 performed minimal physical activity (Group 3-sham). Demographic characteristics, metabolic equivalents (MET), premature ejaculation diagnostic tool (PEDT) and intravaginal ejaculatory latency time (IELT) were recorded. There were no significant differences among three groups in terms of age, BMI, MET, PEDT or IELT before treatment. At the end of the study, there was significant decrease in PEDT scores, and increase in IELT in groups 1 and 2 as compared to Group 3. In conclusion, a moderate physical activity longer than 30 min at least 5 times a week leads to ejaculation delay, and appears as an alternative to dapoxetine on demand for the treatment of PE.
  • Trimetazidine has protective effects on spermatogenesis in a streptozotocin-induced diabetic rat model
    The aim of this study was to investigate the protective effects of trimetazidine (TMZ), as an antioxidant agent, on streptozotocin (STZ)-induced diabetic rats. A total of 50 male Sprague Dawley rats were randomly classified into five groups as follows: Group 1 (control), Group 2 (STZ-induced diabetic rats), Group 3 (STZ-induced diabetic rats treated orally with 1 cc/day isotonic saline), Group 4 (diabetic rats treated orally with 10 mg/kg/day TMZ) and Group 5 (diabetic rats treated orally with 20 mg/kg/day TMZ). After 8 weeks, orchiectomy was carried out. Histopathological and electron microscopic examinations were performed in all groups. In groups 1 and 5, the structural and ultra-structural findings of the testicular tissue and spermatogenesis were found normal. In groups 2, 3 and 4, similar results were obtained in terms of the impaired testicular architecture and degeneration of spermatogenesis. The administration of an optimal dose of TMZ protects against the harmful effects of diabetes mellitus on spermatogenesis in rats. TMZ therapy can be used to maintain normal spermatogenesis in diabetic rats.
  • The presence of human papillomavirus in semen does not affect the integrity of sperm DNA
    It remains unknown whether human papillomaviruses (HPVs) in semen affect sperm DNA integrity. We investigated whether the presence of these viruses in semen was associated with an elevated sperm DNA fragmentation index. Semen samples of 22 normozoospermic patients undergoing infertility treatment, nine fertile donors and seven fertile men with a risk of HPV infection (genital warts or condylomas) were included in the study. The samples were examined by an INNO-LiPA test PCR-based reverse hybridisation array that identifies 28 types of HPVs as simple or multiple infections. Sperm DNA integrity was determined by sperm chromatin dispersion assay (SCD). Our preliminary findings demonstrate an increase in HPV infection in infertile men with respect to fertile men. However, the sperm DNA fragmentation index was not increased in semen containing these viruses.
  • Sperm RNA preparation for transcriptomic analysis: Review of the techniques and personal experience
    In the last 10 years, several approaches, including microarrays, have been applied to investigate sperm transcript levels. However, success using microarray profiling is highly dependent of the quality of the RNA obtained. Therefore, the development of methods that deliver highly purified and intact RNA is of utmost importance. The three steps used to achieve this goal, purification of spermatozoa, RNA extraction and evaluation of RNA quality, are reviewed. Following that review and preliminary experiments, we processed sperm samples from seven normozoospermic men with a combination of gradient centrifugation and somatic cell lysis. RNA was extracted using the NucleoSpin RNA XS kit (Macherey-Nagel) and its purity checked using the BioAnalyzer. Hybridisation was done on Agilent SurePrint G3 Human GE 8 × 60K V2 microarrays. We identified 900 transcripts among the 1000 high abundance sperm transcripts reported in the literature. These genes are known to be involved in several biological processes, notably spermatogenesis, transcription regulation, cell growth and differentiation, sperm motility and capacitation, fertilisation, and embryogenesis. Therefore, our methodology is highly suitable for sperm transcriptomic analyses and can be used, notably, to compare mRNA profiles between fertile and infertile males.
  • Feeding hydroalcoholic extract powder of Lepidium meyenii (maca) enhances testicular gene expression of 3β-hydroxysteroid dehydrogenase in rats
    Although feeding diets containing the extract powder of Lepidium meyenii (maca), a plant growing in Peru's Central Andes, increases serum testosterone concentration associated with enhanced ability of testosterone production by Leydig cells in male rats, changes in testicular steroidogenesis-related factors by the maca treatment are not known. This study examined the effects of maca on testicular gene expressions for luteinizing hormone receptor, steroidogenic acute regulatory protein and steroidogenic enzymes. Eight-week-old male rats were given the diets with or without (control) the maca extract powder (2%) for 6 weeks, and mRNA levels were determined by reverse transcription quantitative real-time PCR. The results showed that the testicular mRNA level of HSD3B1 (3β-hydroxysteroid dehydrogenase; 3β-HSD) increased by the treatment, whereas the levels of the other factors examined did not change. These results suggest that increased expression of 3β-HSD gene may be involved in the enhanced steroidogenic ability by the maca treatment in rat testes.
  • Radiofrequency electromagnetic radiation from cell phone causes defective testicular function in male Wistar rats
    Cell phones have become an integral part of everyday life. As cell phone usage has become more widespread, concerns have increased regarding the harmful effects of radiofrequency electromagnetic radiation from these devices. The current study was undertaken to investigate the effects of the emitted radiation by cell phones on testicular histomorphometry and biochemical analyses. Adult male Wistar rats weighing 180–200 g were randomly allotted to control, group A (switched off mode exposure), group B (1-hr exposure), group C (2-hr exposure) and group D (3-hr exposure). The animals were exposed to radiofrequency electromagnetic radiation of cell phone for a period of 28 days. Histomorphometry, biochemical and histological investigations were carried out. The histomorphometric parameters showed no significant change (p < .05) in the levels of germinal epithelial diameter in all the experimental groups compared with the control group. There was no significant change (p < .05) in cross-sectional diameter of all the experimental groups compared with the control group. Group D rats showed a significant decrease (p ˂ .05) in lumen diameter compared with group B rats. There was an uneven distribution of germinal epithelial cells in groups B, C and D. However, there was degeneration of the epithelia cells in group D when compared to the control and group B rats. Sera levels of malondialdehyde (MDA) and superoxide dismutase (SOD), which are markers of reactive oxygen species, significantly increased (MDA) and decreased (SOD), respectively, in all the experimental groups compared with the control group. Also sera levels of gonadotropic hormones (FSH, LH and testosterone) significantly decreased (p < .05) in groups C and D compared with the control group. The study demonstrates that chronic exposure to radiofrequency electromagnetic radiation of cell phone leads to defective testicular function that is associated with increased oxidative stress and decreased gonadotropic hormonal profile.
  • Sperm DNA fragmentation affects epigenetic feature in human male pronucleus
    To evaluate whether the sperm DNA fragmentation affects male pronucleus epigenetic factors, semen analysis was performed and DNA fragmentation was assessed by the method of sperm chromatin structure assay (SCSA). Human-mouse interspecies fertilisation was used to create human male pronucleus. Male pronucleus DNA methylation and H4K12 acetylation were evaluated by immunostaining. Results showed a significant positive correlation between the level of sperm DNA fragmentation and DNA methylation in male pronuclei. In other words, an increase in DNA damage caused an upsurge in DNA methylation. In the case of H4K12 acetylation, no correlation was detected between DNA damage and the level of histone acetylation in the normal group, but results for the group in which male pronuclei were derived from sperm cells with DNA fragmentation, increased DNA damage led to a decreased acetylation level. Sperm DNA fragmentation interferes with the active demethylation process and disrupts the insertion of histones into the male chromatin in the male pronucleus, following fertilisation.
  • Ascorbic acid reduces redox potential in human spermatozoa subjected to heat-induced oxidative stress
    Oxidation–reduction potential (ORP) is a new measure of oxidative stress. It is a balance between the total available oxidants and reductants. This study measures the efficiency of ascorbic acid (AA) against oxidative stress induced by either heat alone or heat and hydrogen peroxide in sperm suspensions using the MiOXSYS System. Two concentrations of ascorbic acid (400 and 600 μmol/L) were tested against heat- and heat plus hydrogen peroxide-induced oxidative stress in sperm suspensions after 2 and 4 hr of incubation. Sperm motility and static oxidation reduction potential (sORP) were measured at 2 and 4 hr of incubation at three different temperatures. A significant decrease in sORP was observed as a function of AA concentration. The 600 μmol/L AA had more pronounced reduction in sORP compared to 400 μmol/L AA (p = .001). Significant decreases in sperm motility ranging from 4.89% to 14.02% were observed both as a function of incubation time and addition of H2O2 (p < .001). Ascorbic acid is efficacious to reduce heat-induced oxidative stress in sperm preparations in vitro. The supplementation of ascorbic acid may be advantageous for semen preparations in IUI, IVF and ICSI.
  • Response to cooling of pony stallion semen selected by glass wool filtration
    The aim of this study was to compare the sperm separation technique using filtration through glass wool compared with just diluted cooled semen. Eighteen ejaculates were collected from 6 pony stallions of the Brazilian pony breed. Evaluations were done on pH, osmolarity, total motility, membrane functionality (HOST), membrane integrity (CFDA/PI), morphology and mitochondrial viability (MTT) in fresh, 24 and 48 h of cooled semen at 5°C. After dilution, the half of the extended semen was cooled (control group). The other half was cooled after filtration trough glass wool (filtered group). Retained semen was considered the portion of cells that did not transpose glass wool barrier. Total motility from the control, filtered and retained groups after 24 h of cooling was 35.5%, 43.3% and 10% (p < .0001) respectively. Sperm membrane integrity percentage at the CFDA/PI test was 37.9%, 44.8% and 14.8% (p < .0001), on the control, filtered and retained groups respectively. The results confirmed that the passage of spermatozoa through glass wool increased the selection of spermatozoa from pony stallions with higher motility, mitochondrial viability and membrane integrity for cooling in milk extender up to 24 h. Moreover, it was not obtained higher sperm parameters to control after cooling 48 h under the conditions that the study was conducted.
  • Cryoprotectant effect of trehalose in extender on post-thaw quality and in vivo fertility of water buffalo (Bubalus bubalis) bull spermatozoa
    This study was designed to ascertain the cryoprotectant effects of different concentrations of trehalose [0 (T0), 25 (T25), 35 (T35), 45 (T45) mm], egg yolk [20% (E20), 15% (E15) v/v] and glycerol [7% (G7), 5% (G5) v/v] in Tris-citric acid-based extender on post-thaw quality and in vivo fertility of buffalo bull spermatozoa. Twenty-five ejaculates were collected from five bulls and split into four parts. After that, the split ejaculates from each of the bull were diluted either in T0E20G7 (control) or T25E20G5 or T35E15G5 or T45E15G5 extender. Finally, the sperm suspension was frozen in 0.54-ml French straws. Post-thaw sperm total motility (%), progressive motility (%), rapid velocity (%), average path velocity (μm/s), straightline velocity (μm/s), curvilinear velocity (μm/s), linearity (%), plasma membrane and acrosome integrities (%) were higher (p < .05) in T45E15G5 extender as compared to other treatment groups and control. The fertility rate (56.8% versus 41.3%) was higher (p < .05) in buffaloes inseminated with semen doses cryopreserved in extender containing T45E15G5 combination of cryoprotectants than the control. In conclusion, addition of 45 mm trehalose along with 15% egg yolk and 5% glycerol in extender improves the post-thaw quality and in vivo fertility of buffalo bull spermatozoa.
  • Antioxidant treatment ameliorates diabetes-induced dysfunction of the vas deferens in a rat model
    Diabetes mellitus (DM) affects the male ejaculatory function. This study was designed to evaluate the role of oxidative stress in the development of diabetes-induced dysfunction of vas deferens (VD) in the rat. DM was induced by streptozotocin in 40 male Wistar rats. Subsequently, the diabetic animals were divided into three groups: DM group, DM + Eda group and DM + Tau group. These groups were administered saline, edaravone and taurine, respectively, daily for 4 weeks. Another group of ten rats served as a control group. DM was diagnosed in the 40 streptozotocin-injected rats. DM significantly reduced the VD weight. Additionally, DM induced in vitro VD hypercontractility, VD histological abnormalities and increased the serum and VD tissue concentration of malondialdehyde. VD immunohistochemistry revealed overexpression of three markers of oxidative stress. DM significantly reduced serum testosterone levels. No live birth was documented in all DM rats in mating experiments. Antioxidants significantly improved all the aforementioned parameters, except the testosterone levels. This study indicates a deleterious impact of DM-induced oxidative stress on VD histological and functional features. Antioxidant treatment may provide an adjunct tool to alleviate ejaculatory disorders for male patients with type 1 diabetes.
  • Supplementing oregano essential oil to boar diet with strengthened fish oil: Effects on semen antioxidant status and semen quality parameters
    Previous research has shown benefits of dietary fish oil supplementation on semen quality of boars. However, little is known about how antioxidant protects lipid peroxidation on spermatozoa from n-3 polyunsaturated fatty acid (PUFA) addition. This study evaluated the effect of oregano essential oil (OEO) supplementation on semen antioxidant status and semen quality in boars fed a diet enriched with fish oil. Thirty-four mature boars of proven fertility, received daily 2.5 kg basal diet top-dressed with 45 g soybean oil and 15 g fish oil to meet the n-3 PUFA requirement of spermatozoa, randomly allocated to one of four groups supplemented with 100 mg α-tocopheryl acetate kg−1 (control), or 250 or 500 or 750 mg OEO kg−1 for 16 weeks. Semen was collected at weeks 0, 8, 12 and 16 for measurements of sperm production, motion characteristics, sperm α-tocopherol content, antioxidant enzyme activities, reactive oxygen species (ROS), DNA damage (8-hydroxydeoxyguanosine, 8-OHdG), lipoperoxidation (malondialdehyde, MDA) and seminal total antioxidant capacity (TAC). Sperm production and motion characteristics were similar (p > .05) among groups throughout the experimental week 16, but increased (p < .01) with experimental week. Although higher α-tocopherol content and superoxide dismutase (SOD) activities were in OEO group spermatozoa, feeding diet with 500 mg/kg OEO resulted in elevation in seminal TAC, decrease in sperm ROS, MDA and 8-OHdG than control group (p < .05). Overall, these results support the view that oregano essential oil has a positive effect on antioxidant capacity in boar when used fish oil.
  • Assessment of human sperm DNA integrity using two cytochemical tests: Acridine orange test and toluidine blue assay
    Primary infertility affects approximately 15% of couples, with male factor infertility accounting for 50% of cases. Semen samples from 41 patients with asthenoteratospermia and 28 men with proven fertility were analysed according to World Health Organization guidelines. Abnormal sperm chromatin structure was assessed by toluidine blue assay (TBA), and DNA denaturation (DD) was detected by the acridine orange test (AOT). The mean (±SEM) rates of DD and abnormal chromatin structure were significantly higher in infertile subjects compared to fertile group respectively p = .003 and p < .001. A significant correlation was established between sperm DD and abnormal chromatin structure (R = .431, p < .001). Sperm DNA damage correlated significantly with abnormal morphology, sperm motility and necrozoospermia. Our study shows that men with increased levels of abnormal sperm chromatin structure have a high incidence of DNA denaturation and altered semen parameters. These findings suggest that male infertility has been linked to sperm DNA damage.
  • Dietary omega-3 polyunsaturated fatty acids induce plasminogen activator activity and DNA damage in rabbit spermatozoa
    The aim of this study was to determine the effect(s) of dietary omega-3 polyunsaturated fatty acids (ω-3 PUFA) on rabbit semen. Adult rabbit bucks were assigned to two groups that were given two diets, a standard diet (control) and a diet supplemented with ω-3 PUFA. Sperm samples were collected from all bucks with the use of an artificial vagina in 20-day intervals, for a total period of 120 days. The enrichment of membranes in ω-3 PUFA was manifested by the elevation of the 22:5 ω-3 (docosapentaenoic acid [DPA]) levels within 40 days. This increase in DPA content did not affect semen characteristics (i.e., concentration, motility and viability). However, it was associated with the induction of lipid peroxidation in spermatozoa, as determined on the basis of the malondialdehyde content. Lipid peroxidation was associated with DNA fragmentation in ω-3 PUFA-enriched spermatozoa and a concomitant increase in plasminogen activator (PA) activity. The effects of ω-3 PUFA on sperm cells were evident within 40 days of ω-3 PUFA dietary intake and exhibited peack values on day 120. Our findings suggest that an ω-3 PUFA-rich diet may not affect semen characteristics; however, it may have a negative impact on the oxidative status and DNA integrity of the spermatozoa, which was associated with an induction of PAs activity.
  • Ethanol-induced male infertility: Effects of aqueous leaf extract of Tetracarpidium conophorum
    This study investigated the effects of Tetracarpidium conophorum leaf extract on infertility induced by ethanol in male rats. Thirty rats were randomly divided into six groups of five animals each: Group 1 (positive control) received 0.9% saline only; Group 2 (ethanol alone) were given only 30% ethanol orally at 7 ml/kg body weight per day, thrice in a week; groups 3, 4 and 5 were given ethanol and co-treated with 50, 500 and 1,000 mg/kg body weight of leaf extract, respectively, while Group 6 was given ethanol and co-treated with a fertility drug, clomiphene citrate. Ethanol treatment resulted in significant (p < .05) decrease in LDH activity, G-6PDH activity, glycogen content, 3β and 17β HSD activities and testicular and epididymal Zn and Se contents and furthermore decrease in testicular sperm count, viability and marked increment in total sperm abnormalities, rate of sperm analysis parameters and consequently decreased reproductive hormone levels. Interestingly, co-administration of ethanol with either T. conophorum extract or drug almost ameliorated the toxic assault imposed by ethanol on reproductive organs and improved seminal qualities of the rats.
  • In vitro antioxidant effect of curcumin on human sperm quality in leucocytospermia
    Decreased sperm quality was caused by oxidative stress in semen from patients with leucocytospermia. Curcumin is a traditional Chinese herbal monomer extracted from Zingiberaceae turmeric and zedoary turmeric and has antioxidative and anti-inflammatory effects. This study aimed to investigate the effects and specific molecular mechanisms of curcumin on sperm quality in patients diagnosed with leucocytospermia. Forty cases of semen samples were collected from patients with leucocytospermia and 35 cases from normal fertile male. Computer-assisted semen analysis (CASA) was used to detect sperm motility after curcumin incubation. ELISA was used to measure the changes in H2O2, sperm mitochondrial DNA (mtDNA), cytochrome B (Cyt B) and NADH dehydrogenase 5 (NADH5) contents before and after curcumin treatment. The results indicate that curcumin can significantly improve sperm motility from the patients with leucocytospermia. After curcumin treatment, the level of the H2O2 was significantly decreased in the supernatant of curcumin-incubated spermatozoa from leucocytospermic patients. The content of mtDNA was significantly decreased, while the content of Cyt B and NADH5 in spermatozoa was significantly increased. In conclusion, curcumin can significantly improve sperm motility of leucocytospermic patients, against the oxidative damage induced by H2O2. Therefore, curcumin may play a role in mitigating the H2O2-induced injury to sperm.
  • Review on arsenic-induced toxicity in male reproductive system and its amelioration
    Arsenic is an environmental toxicant which causes mutagenic, teratogenic and carcinogenic defects. It is used as herbicide, fungicide and rodenticide and results in contamination of air, soil and water. Arsenic is also produced through burning of coal industries. The sludge of factories contaminates the fodder and drinking source of water of human and livestock. Arsenic binds thiol groups in tissue proteins and impairs the function of the proteins. This metal affects the mitochondrial enzymes and interrupts the production of energy. Oxidative stress and the generation of reactive oxygen species could also be a consequence of arsenic exposure. High arsenic level may suppress the sensitivity of gonadotroph cells to GnRH as well as gonadotropin secretion by elevating plasma levels of glucocorticoids. These ultimately lead to the development of gonad toxicity in animals and cause the reduction in sperm number, sperm viability and motility. Massive degeneration of germ cells and alterations in the level of LH, FSH and testosterone are also reported. The objective of this review was to find out the effects of arsenic-induced toxicity on male reproductive system in animals and its amelioration.
  • Nucleotide composition and codon usage bias of SRY gene
    The SRY gene is present within the sex-determining region of the Y chromosome which is responsible for maleness in mammals. The nonuniform usage of synonymous codons in the mRNA transcript for encoding a particular amino acid is the codon usage bias (CUB). Analysis of codon usage pattern is important to understand the genetic and molecular organisation of a gene. It also helps in heterologous gene expression, design of primer and synthetic gene. However, the analysis of codon usage bias of SRY gene was not yet studied. We have used bioinformatic tools to analyse codon usage bias of SRY gene across mammals. Codon bias index (CBI) indicated that the overall extent of codon usage bias was weak. The relative synonymous codon usage (RSCU) analysis suggested that most frequently used codons had an A or C at the third codon position. Compositional constraint played an important role in codon usage pattern as evident from correspondence analysis (CA). Significant correlation among nucleotides constraints indicated that both mutation pressure and natural selection affect the codon usage pattern. Neutrality plot suggested that natural selection might play a major role, while mutation pressure might play a minor role in codon usage pattern in SRY gene in different species of mammals.
  • Oxidative stress status and sperm DNA fragmentation in fertile and infertile men
    Evidence suggests that disturbing the balance between reactive oxygen species levels and antioxidant contents in seminal plasma leads to oxidative stress resulting in male infertility. This study was carried out to identifying clinical significance of seminal oxidative stress and sperm DNA fragmentation in treatment strategies of male infertility in southwest Iran. Sperm parameters, lipid peroxidation and activity of antioxidant enzymes were assessed in fertile (n = 105) and infertile (n = 112) men. Malondialdehyde (MDA) levels in seminal plasma were found to be higher significantly (p < .001) in patients. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities in seminal plasma were significantly (p < .001) lower in infertile men. Significant negative correlations were observed between MDA levels and sperm motility and normal morphology. Spermatozoa with fragmented DNA were higher (p < .001) in infertile men and significantly correlated with MDA levels and SOD and GPx activities. MDA of 4.2 nmol/ml, SOD of 4.89 U/ml and GPx of 329.6 mU/ml were optimum cut-off limits to discriminate infertile patients from fertile men. The results show the leading role of oxidative stress in aetiology of male infertility in southwest Iran and indicate that evaluation of seminal antioxidant status and DNA integrity can be helpful in men attending infertility clinics during fertility assessment.
  • Quantitative changes in testicular structure and function in rat exposed to mobile phone radiation
    The possible effects of the electromagnetic fields (EMF) generated by mobile phones on reproductive functions have been discussed in recent years. The aim of this study was to evaluate the effects of EMF emitted from mobile phones on the rat testis morphology and histopathology using stereological techniques. We also investigated cortisol, testosterone, FSH and LH levels. A total of thirty-two (n = 32) male Wistar albino rats were used in this study. Animals were randomly divided into four groups as control (C, n = 8), sham (Sh, n = 8), mobile phone speech (Sp, n = 8) and mobile phone standby (ST by). Morphometric measurements were made with the help of a computer-assisted stereological analysis system. The testis weight and volume were significantly lower in the EMF exposed groups. The mean volume fraction of interstitial tissue was higher, but the volume fraction of tubular tissue was lower in the EMF-exposed groups. The mean tubular and germinal tissue volume, seminiferous tubule diameter and germinal epithelium height were also lower in EMF exposed groups. The cortisol levels in the EMF-exposed groups were significantly higher. In conclusion, the EMF created by mobile phones caused morphologic and histological changes by the affecting germinal epithelium tissue negatively.
  • Metabonomic analysis of fatty acids in seminal plasma between healthy and asthenozoospermic men based on gas chromatography mass spectrometry
    The mechanism of asthenozoospermia remains unclear. The knowledge of the metabolism of fatty acids in seminal plasma is important and meaningful for the pathological study of asthenozoospermia. We present an optimised assay of extraction and derivatisation followed by GC/MS to analyse metabolites, especially fatty acids, in seminal plasma from healthy and asthenozoospermic men. Eighty-nine peaks including 17 kinds of fatty acids were analysed and identified in the chromatogram. The GC/MS data were analysed using t test, fold change and partial least squares discriminant analysis to explore the potential biomarkers of asthenozoospermia. Seven metabolites in asthenozoospermic group were found to be significantly different from those in the normal group (with p < .05, fold change >1.2 and variable importance for projection >1). Of which, high levels of oleic acid and palmitic acid in seminal plasma from asthenozoospermic men may indicate a membrane metabolism disorder in spermatozoa and the lack of valine in the asthenozoospermic group may contribute to poor sperm motility. The results may facilitate the understanding of the role of fatty acids and amino acids in asthenozoospermia and provide solid foundation for further pathological study of asthenozoospermia.
  • 6-Gingerol-rich fraction prevents disruption of histomorphometry and marker enzymes of testicular function in carbendazim-treated rats
    Previous investigations demonstrated that 6-gingerol-rich fraction (6-GRF) prevented testicular toxicity via inhibition of oxidative stress and endocrine disruption in CBZ-treated rats. The influence of 6-GRF on alterations in histomorphometry and marker enzymes of testicular function in CBZ-treated rats which hitherto has not been reported was investigated in this study. The animals were orally administered either CBZ (50 mg/kg) alone or in combination with 6-GRF (50, 100 and 200 mg/kg) for 14 consecutive days. Histomorphormetric analysis demonstrated that 6-GRF significantly prevented CBZ-mediated increase in the organo-somatic index of the testes and seminiferous tubular diameter as well as the reduction in epithelium height and tubular length of testes in the rats. Similarly, 6-GRF ameliorated CBZ-induced disruption in the epithelium height as well as in the proportion of tubule and interstitium of the epididymis the treated rats. Furthermore, 6-GRF prevented CBZ-mediated increase in testicular acid phosphatase activity and the decrease in testicular alkaline phosphatase, aminotransferases, glucose-6-phosphate dehydrogenase and lactate dehydrogenase activities. Moreover, 6-GRF ameliorated CBZ-induced reduction in the testicular and epididymal sperm count and sperm motility in the treated rats. Conclusively, 6-GRF enhances key functional enzymes involve in spermatogenesis and maintains histo-architecture of testes and epididymis in CBZ-treated rats.
  • Use of the fluorescent dye tetramethylrhodamine methyl ester perchlorate for mitochondrial membrane potential assessment in human spermatozoa
    Mitochondrial membrane potential (ΔΨm) is an indicator of sperm quality and its evaluation complements the standard semen analysis. The fluorescent dye JC-1 has been widely used to assess sperm ΔΨm; however, some problems have been detected under certain experimental conditions. Another fluorescent compound, tetramethylrhodamine methyl ester perchlorate (TMRM), has been used in somatic cells and bovine spermatozoa but not in human spermatozoa. TMRM accumulates in hyperpolarised mitochondria and the fluorescence intensity of this compound correlates with ΔΨm. Thus, the aim of this study was to evaluate and validate the usefulness of the fluorescent dye TMRM for measuring sperm ΔΨm. The results showed that TMRM accurately detects sperm populations displaying either high or low ΔΨm. Moreover, TMRM was able to measure sperm ΔΨm under the experimental conditions in which JC-1 had previously presented difficulties. Differences in ΔΨm according to sperm and semen quality were properly detected and a positive correlation between ΔΨm and conventional semen parameters was observed. Finally, a positive correlation was found between the ΔΨm measurement by TMRM and by the widely used JC-1. In conclusion, TMRM is a simple, time-effective method, easy to set in laboratories equipped with flow cytometry technology, and can accurately detect changes in ΔΨm with efficiency comparable to JC-1 without its limitations.
  • Relationship between Chlamydia trachomatis and Mycoplasma genitalium infection and pregnancy rate and outcome in Iranian infertile couples
    The study was performed to investigate the prevalence of Chlamydia trachomatis and Mycoplasma genitalium in a population of infertile couples from Iran and how this relates to tubal factor infertility, pregnancy rate and outcome of pregnancy. Blood, semen and first-void urine samples were obtained from 250 infertile couples and 250 fertile women as a control. Infertile couples were followed up after 24 months to determine diagnosis, referral for assisted conception, any pregnancy and pregnancy outcome. Data were analysed with regard to the results of (i) serological analysis for specific antibodies to C. trachomatis in serum; (ii) the presence of C. trachomatis and M. genitaliumDNA in first-void urine; and (iii) in a semen sample of the male partner. Prevalence of C. trachomatis in our study population was comparable to other studies using similar methods and test specimens. No evidence of M. genitalium infection was found. Detection of C. trachomatis in one partner rarely correlated with infection in the other. The risk of tubal factor infertility and the probability of pregnancy and pregnancy outcome were unrelated to the results of serological tests for C. trachomatis antibodies or the presence of C. trachomatisDNA in first-void urine of both partners and in a semen sample provided by the male.
  • Relative contributions of testosterone deficiency and metabolism syndrome at the risk of reduced quality of life: A cross-sectional study among Chinese mid-aged and elderly men
    Testosterone deficiency and metabolism syndrome (MetS) are universal among ageing males, and they have been suggested responsible for poorer quality of life (QoL). We aimed to evaluate the relative contributions of reproductive hormones and components of MetS at the risk of reduced QoL among Chinese mid-aged and elderly men. A cross-sectional study recruited 2,364 males aged 40–79 years, and 2,165 was included for analysis eventually. The Chinese version of ageing male symptoms scale, 36-item Short Form and Beck Depression Inventory were applied to assess QoL. Bivariate correlation analysis and multiple linear regression analysis were used to assess the relative contributions of reproductive hormones and components of MetS at the risk of reduced QoL. Testosterone deficiency and MetS contributed to poorer QoL, of which higher fasting blood glucose made the primary contribution, lower total testosterone mainly contributed to poorer physical functioning.
  • Effects of treatment with Hypoxis hemerocallidea extract on sexual behaviour and reproductive parameters in male rats
    Hypoxis hemerocallidea is used in traditional medicine in South Africa, for the treatment of male reproductive ailments and various chronic illnesses. Despite chronic use, its effects on male reproductive system are unknown. Male Wistar rats were treated orally daily for 28 (n = 18) and 56 days (n = 18). Treatment groups (n = 6/group) per treatment period were as follows: untreated control, 150 mg/kg and 300 mg/kg 70% ethanolic extract of H. hemerocallidea. Sexual behaviour observations were performed on days 17 and 42 of the study. Sperm, biochemical and testicular histopathological studies were carried out. Arousal and libido and serum testosterone increased after 56 days of treatment. There was an increase in epididymal sperm count at both treatment doses, with the 300 mg/kg dose showing a higher sperm count (p < .05) compared to the 150 mg/kg treatment group. The higher 300 mg/kg dose also showed an increase (p < .05) in sperm motility after 56 days of treatment. Histology showed an increase in germinal layer thickness, consistent with the observed increase in sperm count. Testicular oxidative status improved after 56 days of treatment. Results suggest that chronic treatment with H. hemerocallidea may improve male sexual function and fertility parameters and may protect testes from oxidative damage.
  • Decrease in bovine in vitro embryo production efficiency during winter season in a warm-summer Mediterranean climate
    Retrospective analysis of monthly embryo production from December 2011 to May 2015 and its correlation with meteorological data in our geographic zone was made. We had observed that in certain time of the year, in vitro blastocyst production decreases. Accordingly, was examined the association between blastocyst production and climatological parameters. Cleavage rates correlate positively with blastocyst rates (p < .05). Significant differences in cleavage rates between autumn and summer (79.8%; 71.5%), and between winter and autumn (71.8%; 79.8%), were found. Blastocyst production had lower efficiency in June (9 ± 12%) and July (4.9 ± 5.7%), which coincides with winter season. In contrast, higher embryo production was obtained in February (22.2 ± 9.7%), March (22.9 ± 14%) and September (25.2 ± 6.6%), which coincides with autumn and spring season. Similarly, embryo production correlates with meteorological parameters: blastocyst production positively correlates with sunshine hours, maximum temperature and average temperature. Similarly, blastocyst production inversely correlates with total precipitation and days >1 mm precipitation (p < .05). There is a significant decrease in bovine in vitro embryo production efficiency during winter season in our warm-summer Mediterranean climate zone. It remains to be investigated the direct effect of environmental factors on oocyte quality and its impact on in vitro production efficiency.
  • In vitro reprotoxicity of carboxyl-functionalised single- and multi-walled carbon nanotubes on human spermatozoa
    Reproductive toxicity of carboxyl-functionalised carbon nanotubes (CNT-COOH), as the most commonly used form of water-soluble CNTs, is not clearly studied. The aim of this study was to investigate in vitro toxicity of carboxylated single-walled and multi-walled CNTs (SWCNT-COOH and MWCNT-COOH) against human spermatozoa. Sperm cells from healthy donors were incubated with 0.1–100 μg/ml of SWCNT-COOH or MWCNT-COOH at 37°C for up to 5 hr. Viability of sperm cells was assessed using MTT test, and sperm motility was evaluated following World Health Organization guideline. Production of reactive oxygen species (ROS) and nitric oxide (NO) in sperm was also assessed. We showed that both MWCNT-COOH and SWCNT-COOH following incubation in vitro with human spermatozoa did not exert negative effect on viability while motility was significantly (p < .05) dropped in a dose-dependent manner. Moreover, there was no significant effect of the type, dose and exposure time of the CNT-COOH on NO production. Exposure of sperm cells to both examined types of CNTs at concentrations as low as 0.1 μg/ml caused a significant increase in ROS levels. In conclusion, carboxylated forms of CNTs seem to be harmful for human spermatozoa. Further studies, especially using in vivo models, are needed to decide about reprotoxicity of carboxylated forms of CNTs.
  • A potential mechanism associated with lead-induced testicular toxicity in rats
    This study was conducted to investigate the mechanism of lead (Pb)-induced testicular toxicity. We examined the impact of Pb toxicity on 17β-oestradiol (E2), oestrogen receptors (ERs) and aromatase P450 which are key factors in spermatogenesis. Treatment of rats with Pb acetate (PbAc, 50 mg/L in drinking water) significantly reduced sperm count, motility, viability and increased sperm abnormalities along with degenerative changes in seminiferous tubules and Leydig cells. Additionally, administration of PbAc resulted in a significant reduction in serum testosterone, serum and testicular E2 as well as increased level of testicular testosterone. Pb also induced testicular oxidative stress as evidenced by a significant decrease in the activities of superoxide dismutase, glutathione peroxidase and catalase antioxidant enzymes, and increased malondialdehyde level in the testis. At the molecular level, Pb treatment downregulated the mRNA expression of P450 arom (Cyp19) and ERα. In conclusion, Pb induces testicular oxidative damage and disrupts spermatogenesis, at least in part, via downregulation of Cyp19 and ERα expression, which further decrease E2 level. These data, therefore, provide insight into the mechanism of lead-induced testicular toxicity.
  • Evaluation of serum level of Osteocalcin hormone in male infertility
    The suggested concept of “bone as an endocrine organ” had shed the light on the role of osteocalcin, an osteoblast secreted hormone, in regulation of testosterone production. This study aimed to assess the association between the active undercarboxylated form of osteocalcin (ucOC) and semen parameters and hormonal levels in infertile male patients. The study was carried on 34 infertile male patients and 20 fertile healthy control males. Semen analysis and serum level of testosterone, LH and FSH were performed in addition to serum level of ucOC in cases and controls. The results revealed significant differences between cases and controls in all measured semen and hormonal parameters. In addition, significant higher level of ucOC in cases than control group (p = .019). On the other hand, ucOC was not related significantly to any of the measured hormones or semen parameters. There was no significant correlation between ucOC and sperm concentration, total motility, morphology (p = .594, .640, .940 respectively) and similarly between ucOC and testosterone level or LH level (p = .275, .954 respectively). The significant higher level of ucOC in infertile cases cannot be used as a predictor of male reproductive parameters.
  • Histopathological and biochemical comparisons of the protective effects of amifostine and l-carnitine against radiation-induced acute testicular toxicity in rats
    The aim of this study was to compare the radioprotective efficacies of amifostine (AMI) and l-carnitine (LC) against radiation-induced acute testicular damage. Thirty Wistar albino rats were randomly assigned to four groups: control (n = 6), AMI plus radiotherapy (RT) (n = 8), LC plus RT (n = 8) and RT group (n = 8). The rats were irradiated with a single dose of 20 Gy to the scrotal field. LC (300 mg/kg) and AMI (200 mg/kg) were given intraperitoneally 30 min before irradiation. The mean seminiferous tubule diameters (MSTDs) were calculated. Testicular damage was evaluated histopathologically using Johnsen's mean testicular biopsy score criteria. Malondialdehyde (MDA) and glutathione levels were measured in tissue samples. AMI plus RT and LC plus RT groups had significantly higher MSTDs than those in the RT group (p = .003 and p = .032 respectively). MDA values of both AMI plus RT and LC plus RT groups were significantly lower than those in RT group (p < .004 and p < .012 respectively). As a result, AMI and LC have a similar radioprotective effect against radiation-induced acute testicular damage, histopathologically and biochemically.
  • Does detection of DDX4 mRNA in cell-free seminal plasma represents a reliable noninvasive germ cell marker in patients with nonobstructive azoospermia?
    This study aimed to investigate the potential application of DDX4 gene expression in cell-free seminal mRNA as a noninvasive biomarker for the identification of the presence of germ cells in men with nonobstructive azoospermia and to correlate this factor with testicular biopsy. Male reproductive organ-specific genes were chosen: DDX4, which is a germ cell-specific gene and transglutaminase 4, which is a prostate-specific gene that was used as a control gene. Thirty-nine azoospermic males and twenty-eight normospermic fertile males (serving as a control group) participated in the study. Histopathological examination of testicular biopsies categorised azoospermic males into 20.5% with maturation arrest, 17.9% with incomplete Sertoli cell-only syndrome and 61.5% with complete Sertoli cell-only syndrome. Using real-time polymerase chain reaction, positivity for DDX4 gene was detected in 17 of 39 males with NOA which was due to maturation arrest in 35.3% (n = 6/17) of cases, due to incomplete Sertoli cell only in 23.5% (n = 4/17) and due to complete Sertoli cell only in 41.2% (n = 7/17). The study recommends joint utilisation of molecular transcripts as noninvasive biomarkers and histopathological examination of testicular biopsies in management of cases with azoospermia of the nonobstructive type.
  • Blood and semen paraoxonase—arylesterase activities in normozoospermic and azoospermic men
    Paraoxonase and arylesterase enzymes are corner stones of antioxidant defence. We aimed to compare azoospermic infertile men and normozoospermic individuals with respect to total antioxidant status (TAS), total oxidant status (TOS), oxidative stress index (OSI), paraoxonase and arylesterase levels in the blood and seminal plasma. Two-hundred consecutive infertility patients and voluntarily participated were included. In the normozoospermic group, TAS, PON, arylesterase values were statistically significantly higher when compared with those in the azoospermic group, while lower TOS and OSI levels were observed in the blood and seminal plasma of azoospermic group. In the semen analyses of normozoospermic group, the correlation between semen volume, sperm concentration, sperm motility and morphology and TAS, TOS, OSI, PON and arylesterase values was examined. A negative correlation was determined between semen volume and OSI. Levels of serum oxidative parameters were higher in the azoospermic group relative to normozoospermic group, but antioxidant parameters were lower than those of the normozoospermic group. Oxidative stress performs an essential role in the aetiology of male infertility by negatively influencing sperm quality and function. Assessment of blood and seminal plasma oxidative profiles might be an important tool to better evaluation of sperm reproductive capacity and functional competence.
  • Pentoxifylline besides naltrexone recovers morphine-induced inflammation in male reproductive system of rats by regulating Toll-like receptor pathway
    This study aimed to investigate the effect of pentoxifylline on complications of prolonged usage of morphine upon the testis and sperm parameters of rats. In this study, forty male Wistar rats were divided into five groups (n = 8) and treated for 56 days to only saline, only morphine, only pentoxifylline, pentoxifylline + morphine and naltrexone + morphine. The diameters of seminiferous tubules, the maturity of germ line epithelium and sperm parameters were evaluated. The expression of inflammatory-related factors in testis tissues were also investigated at gene and protein levels. The data were calculated by one-way ANOVA test followed by Tukey's post hoc test using SPSS software for windows (version 20). Seminiferous tubule diameter, the maturity of spermatogonia and sperm parameters were significantly decreased in morphine group in comparison with control, pentoxifylline and pentoxifylline + morphine groups (p < .001). The expression of anti-inflammatory markers, at both gene and protein levels, was significantly increased in testis of morphine-treated rats in comparison with other groups (p < .001). Chronic morphine administration induces destructive effects on male reproductive system by regulating inflammatory responses. Pentoxifylline recovers the destructive effects of morphine on male reproductive system by inhibiting TLR (Toll-like receptor) activity, as an anti-inflammatory response.
  • Correlation of genetic results with testicular histology, hormones and sperm retrieval in nonobstructive azoospermia patients with testis biopsy
    To investigate the frequency and types of genetic results in different testicular histology of patients with nonobstructive azoospermia (NOA), and correlated with hormones and sperm retrieval (SR), a retrospective study was conducted in 286 Chinese NOA patients who underwent testis biopsy and 100 age-matched fertile men as the control group. Chromosome karyotype analyses were performed by the peripheral blood chromosome G-band detection method. Screening of Y chromosome microdeletions of azoospermia factor (AZF) region was performed by polymerase chain reaction (PCR) amplification of 11 sequence-tagged sites (STS). The serum levels of follicle-stimulating hormone, luteinising hormone and testosterone (T) and the appearance of scrotal ultrasound were also obtained. In 286 cases of NOA, 14.3% were found to have chromosomal alterations. The incidence of chromosomal abnormality was 2.8%. Sex chromosomal abnormalities were seen in six cases (four cases of Klinefelter's syndrome (47, XXY) and two cases of mosaics). The incidence of polymorphic chromosomal variants was 3% in the normal group and 11.5% in the NOA group. In total, 15.7% of NOA patients were found to have AZF microdeletions and AZF (c + d) was the most frequent one. The results of hormone and SR were found to be significantly different among all testicular histological types, whereas no significant differences were found when it comes to genetic alterations. It is concluded that the rate of cytogenetic alterations was high in NOA patients. So screening for chromosomal alterations and AZF microdeletions would add useful information for genetic counselling in NOA patients with testis biopsy and avoid vertical transmission of genetic defects by assisted reproductive technology.
  • Ameliorating effects of fennel and cumin extracts on sperm quality and spermatogenic cells apoptosis by inducing weight loss and reducing leptin concentration in diet-induced obese rats
    This study was established a model of obesity to estimate the impact of fennel and cumin as anti-obesity extracts on body weight, body mass index (BMI), food consumption, leptin concentration, sperm quality and testis architecture to determine the reversibility of reproductive function of obese animals. Male rats were randomly assigned to either a normal or high-fat diet for 8 weeks. Then, we divided 56 adult rats into seven groups: control (CO); obesity (OB); fennel 100 and 200 mg/kg; cumin 50 and 100 mg/kg; and fennel 100 mg/kg plus cumin 50 mg/kg. From weeks 9-16, the animals treated extracts by gavages daily. We analysed leptin concentration, sperm quality and apoptosis of testis along with evaluating changes in body weight. Body weight of animals increased 25% at week 8. However, body weight, BMI, leptin concentration and apoptosis indices of OB rats increased at the end of study. However, the relative sperm parameters decreased. Nevertheless, fennel and cumin treatment improved sperm quality, and spermatogenic cells apoptosis following weight loss. Concomitant with weight loss, leptin concentration and food consumption decreased. In conclusion, fennel and cumin as supplements may ameliorate sperm quality of obese animals following weight loss and reduction in leptin concentration.
  • Issue Information
  • Is the use of plants in Jordanian folk medicine for the treatment of male sexual dysfunction scientifically based? Review of in vitro and in vivo human and animal studies
    Male sexual dysfunction is a serious problem which has an impact on the quality of life. In Jordanian folk medicine, 56 plant species were reported to be used by males to improve sexual potency and as aphrodisiacs. The aim of this study was to search for scientific evidence justifying their folk use. Of the 15 studied plants, only five were found to enhance spermatogenesis. The other 10 were reported to decrease spermatogenesis at least by one study. The majority of the studied plants possessed a protective effect on testis in different in vivo models as well as antioxidant activities. The effect of these plants on steroidogenesis and the hypothalamic–gonadal axis was also reviewed. The effect of only five plants was studied on sexual behaviour enhancement and three of them were active. Three of the four studied plants enhanced erection. The mechanism of action of active constituents isolated from the studied plants was also investigated. In conclusion, many plants used in Jordanian folk medicine decreased or had no effect on spermatogenesis in animal models. These plants have antioxidant and/or adaptogenic effects, and this may result in a beneficial action on male reproductive system.
  • Nitrosative stress by peroxynitrite impairs ATP production in human spermatozoa
    The most toxic species in live systems include reactive nitrogen species such as peroxynitrite, which at high levels induces nitrosative stress. In human spermatozoa, the negative effect of peroxynitrite on motility and mitochondrial membrane potential was recently demonstrated, and the hypothesis of this work is that impairment of ATP production could be one cause of the effect on motility. Therefore, the aim here was to evaluate ATP production by both glycolysis and oxidative phosphorylation (OXPHOS) in spermatozoa exposed to peroxynitrite in vitro. Human spermatozoa were incubated with SIN-1, a molecule which generates peroxynitrite, and the ATP level was evaluated. Then, to inactivate glycolysis or OXPHOS, spermatozoa were incubated with pharmacological inhibitors of these pathways. Spermatozoa treated for inactivating one or the other pathway were exposed to SIN-1, and the ATP level was compared to the control without SIN-1 in each condition. The ATP level fell after peroxynitrite exposure. The ATP in spermatozoa treated for inactivating one or the other metabolic pathway and subsequently exposed to peroxynitrite was reduced compared with the control. These results show for the first time that an important mechanism by which peroxynitrite reduces sperm function is the inhibition of ATP production, affecting both glycolysis and OXPHOS.
  • Relationship between volume of the seminal vesicles and sexual activity in middle-aged men
    The relationship between volume of the seminal vesicles and the frequency of sex and sexual function in middle-aged men is not clear. This study included 81 patients who were diagnosed with localized prostate cancer. Volume of the seminal vesicles was examined using a volume analyser from computed tomography. Sexual function was subjectively evaluated using the Expanded Prostate Cancer Index Composite and Erection Hardness Score. The frequency of sex was surveyed using our original questionnaire. The mean ± SD age of the patients was 67.7 ± 5.3 years. There was no relationship between the volume of seminal vesicles and age of the patients. Volume of the seminal vesicles in patients who answered that they had sexual activity at least once a year was significantly larger than in those who answered no sexual activity for several years (P < .01) Moreover, among sexually active, middle-aged men, volume of the seminal vesicles was significantly larger in those who had a sexual frequency once every 3 months than in those who had a sexual frequency once every 6 months or once a year (P < .05). Our study suggests that the volume of seminal vesicles of middle-aged men is correlated with sexual activity.
  • Age is a significant predictor of early and late improvement in semen parameters after microsurgical varicocele repair
    Accumulating evidence indicates that varicocele repair improves sperm quality. However, longitudinal changes in sperm parameters and predictors of improved semen characteristics after surgery have not been fully investigated. We retrospectively reviewed data from 100 men who underwent microsurgical subinguinal varicocele repair at a single centre. Follow-up semen examinations were carried out at 3, 6 and 12 months post-operatively. Logistic regression was used to identify predictors of early (3 months) and late (≥6 months) improvement in semen parameters after varicocele repair. At 3 months post-operatively, 76.1% of the patients had improved total motile sperm counts, which continued to improve significantly up to 12 months post-operatively (p = .016). When comparing changes in semen parameters between younger (<37 years) and older (≥37 years) men, post-operative improvements in sperm concentration and motility were greater among younger men. Multivariate analysis showed that younger age was associated with early (p = .043) and late (p = .010) post-operative improvement in total motile sperm count. Our findings indicate that early varicocele repair improved semen parameters after surgery.
  • Co-administration of ginseng and ciprofloxacin ameliorates epididymo-orchitis induced alterations in sperm quality and spermatogenic cells apoptosis following infection in rats
    Korean red ginseng (KRG) may be a beneficial adjuvant along with ciprofloxacin to ameliorate devastating effects of epididymo-orchitis (EO) on male fertility. This study intends to assay the effects of KRG and ciprofloxacin on sperm quality and spermatogenic cells apoptosis in EO rats. We divided 54 adult rats into nine groups (n = 6 rats per group): control (CO), sham-operated (SH), EO (E); ciprofloxacin (C), EO–ciprofloxacin (EC), KRG (G), EO–KRG (EG), ciprofloxacin–KRG (CG) and EO–ciprofloxacin–KRG (ECG). We administered ciprofloxacin and KRG 48 hr after the Escherichia coli (E. coli) injection for 10 days. Bilateral orchiectomy was performed after one sperm cycle (14 days) following the last treatment with ciprofloxacin and KRG. Total and progressive motility of E, C and EC groups decreased. However, motility is improved in CG and ECG in comparison with these groups. The E group induced negative changes in the architecture of testes tissue and dramatic increase in apoptosis indices. Interestingly, co-administration of ciprofloxacin and KRG has dramatically improved Miller's and Johnsen's scores and decreased the apoptosis indices of animals in the ECG group. Combined treatment of ciprofloxacin and KRG may improve the quality of spermatozoa and attenuated apoptosis indices in the ECG group.
  • Testosterone and trenbolone enanthate increase mature myostatin protein expression despite increasing skeletal muscle hypertrophy and satellite cell number in rodent muscle
    The androgen-induced alterations in adult rodent skeletal muscle fibre cross-sectional area (fCSA), satellite cell content and myostatin (Mstn) were examined in 10-month-old Fisher 344 rats (n = 41) assigned to Sham surgery, orchiectomy (ORX), ORX + testosterone (TEST; 7.0 mg week−1) or ORX + trenbolone (TREN; 1.0 mg week−1). After 29 days, animals were euthanised and the levator ani/bulbocavernosus (LABC) muscle complex was harvested for analyses. LABC muscle fCSA was 102% and 94% higher in ORX + TEST and ORX + TREN compared to ORX (p < .001). ORX + TEST and ORX + TREN increased satellite cell numbers by 181% and 178% compared to ORX, respectively (p < .01), with no differences between conditions for myonuclear number per muscle fibre (p = .948). Mstn protein was increased 159% and 169% in the ORX + TEST and ORX + TREN compared to ORX (p < .01). pan-SMAD2/3 protein was ~30–50% greater in ORX compared to SHAM (p = .006), ORX + TEST (p = .037) and ORX + TREN (p = .043), although there were no between-treatment effects regarding phosphorylated SMAD2/3. Mstn, ActrIIb and Mighty mRNAs were lower in ORX, ORX + TEST and ORX + TREN compared to SHAM (p < .05). Testosterone and trenbolone administration increased muscle fCSA and satellite cell number without increasing myonuclei number, and increased Mstn protein levels. Several genes and signalling proteins related to myostatin signalling were differentially regulated by ORX or androgen therapy.
  • Effects of glycerol, equilibration time and antioxidants on post-thaw functional integrity of bovine spermatozoa directly obtained from epididymis
    This work aimed to evaluate the effect of stabilisation times, glycerol concentration, and the catalase and superoxide dismutase supplementation of diluent on parameters of frozen-thawed spermatozoa from epididymis of Nelore bulls: Experiment 1: spermatozoa diluted in Tris-egg yolk with glycerol (3%, 5% or 7%) and stabilisation times (0, 2 or 4 hr at 5°C); Experiment 2: Tris-egg yolk only, Tris-egg yolk with catalase (CAT, 50 or 100 U ml−1) or superoxide dismutase (SOD, 50 or 100 U ml−1). Frozen-thawed spermatozoa were evaluated for kinetic parameters, plasma membrane and acrosome integrity, mitochondrial activity and IVF capacity. ALH and BCF were affected (p < .05) by glycerol at 3% after 4-hr equilibration time and 7% after 2-hr equilibration time. Glycerol 3% had lower (p < .05) iPM and iAc after 4 hr. Glycerol 5% had greater (p < .05) hPMM after 4 hr and iAc after 2 hr than at 0 hr. SOD 100 U ml−1 had lower (p < .05) linearity and wobble compared to control group. No was observed differences to fertilisation rate (p < .05) among groups. In conclusion, glycerol 5% in Tris-egg yolk extender for 4 hr is suitable for the preservation of sperm kinetics and membrane integrity. CAT (50 and 100 U ml−1) or SOD (50–100 U ml−1) had no beneficial effects on sperm kinetics, plasma and acrosomal membrane integrity, mitochondrial activity or the capacity for IVF of frozen-thawed spermatozoa from epididymis of Nelore bulls.
  • Nutrient patterns and asthenozoospermia: a case–control study
    The association of dietary nutrient patterns and sperm motility is not yet well elucidated, and previous studies have just focused on the isolated nutrients. This case–control study examined the association of nutrient patterns with asthenozoospermia among Iranian men. In total, 107 incident asthenozoospermic men and 235 age-matched controls were interviewed through the infertility clinics in Tehran, Iran, from January 2012 to November 2013. Semen quality data were analysed according to the fifth edition of WHO guideline. Nutrient patterns were identified using principal component analysis based on semiquantitative 168-item food frequency questionnaires. All nutrient intakes were energy-adjusted by the residual method. In principal component analysis, three dietary patterns emerged. The first pattern, which was high in vitamin E, vitamin D, vitamin C, zinc, folate, total fibre, selenium and polyunsaturated fatty acids, was significantly associated with lower risk of asthenozoospermia. After adjustment for potential confounders, participants in the highest tertile of the first pattern scores had 51% lower risk of asthenozoospermia compared with those in the lowest (p-trend: .004). Our findings suggest that adherence to the pattern comprising mainly of antioxidant nutrients may be inversely associated with asthenozoospermia.
  • Assessment of seminal mast cells in infertile men with varicocele after surgical repair
    This study aimed to assess seminal mast cells in infertile men associated with varicocele (Vx) pre- and post-surgical repair. Forty-five infertile men associated with Vx were subjected to history taking and clinical examination. In addition, semen parameters and seminal mast cells stained with 1% toluidine blue were estimated pre-varicocelectomy and three months post-varicocelectomy. Vx surgical repair revealed a significant improvement in the mean sperm concentration, progressive sperm motility, total sperm motility and sperm abnormal morphology and a significant decrement in seminal mast cells (mean ± SD, 3.56 ± 2.23 cells per high-power field (HPF) vs. 2.22 ± 1.06 cells per HPF, p = .01). The pre-operative mean mast cell count demonstrated significant increases in cases with Vx grade III compared with other Vx grades and in cases with bilateral Vx compared with unilateral Vx cases. Seminal mast cells demonstrated a significant correlation with sperm concentration, progressive sperm motility and total sperm motility and a nonsignificant correlation with age and sperm abnormal morphology. It is concluded that seminal mast cells decrease significantly in infertile men with Vx after surgical repair showing a significant negative correlation with sperm concentration, progressive sperm motility and total sperm motility.
  • Sperm mitochondrial DNA deletion in Iranian infertiles with asthenozoospermia
    Asthenozoospermia is an important cause of male infertility. The mutations in sperm mitochondrial DNA (mtDNA) result in either functionless or malfunctioning some proteins, subsequently affecting sperm motility leading to asthenozoospermia. The purpose of this study was to investigate sperm mtDNA 4,977-bp deletion in infertile men with low sperm motility/immotile spermatozoa compared to healthy subjects with high sperm motility. Semen samples of 256 asthenozoospermic infertiles and 200 controls from northern Iran were collected. After extraction of spermatozoa total DNA, Gap-polymerase chain reaction (Gap-PCR) was performed. The deletion was observed in 85.93% of patients with asthenozoospermia compared with 14% in controls [OR = 37.5397, 95% confidence interval = 12.937–108.9276, p < .0001]. It is concluded that there is a strong association between sperm mtDNA 4,977-bp deletion and asthenozoospermia-induced infertility in the population examined. Large-scale mtDNA deletions in spermatozoa may induce bioenergetic disorders. Nevertheless, to validate our results broader research may be needed.
  • Significance of platelet distribution width as a severity marker of erectile dysfunction
    Mean platelet volume (MPV) and Platelet distribution width (PDW) are potential markers in platelet activation. In present study, we aimed to evaluate MPV and PDW as potential severity markers for those patients who are complaining erectile dysfunction (ED). A total of 358 participants were enrolled in this study. The whole cohort was asked to complete the International Index of Erectile Function-5 (IIEF-5) questionnaire. The participants were classified into 3 groups: control group (n = 120), mild ED (n = 118) and severe ED (n = 120). We found in our cohort MPV and PDW were significantly higher in both mild ED group and severe ED group than control group (9.24 ± 0.70 and 9.71 ± 0.80 versus 8.56 ± 0.62 for MPV; 14.48 ± 1.29 and 14.98 ± 1.60 versus 12.86 ± 1.13 for PDW respectively). The MPV and PDW increased as the disease progressed. In the mild and severe ED groups, a significant inverse correlation was detected between the mean values of IIEF-5 score and PDW. Furthermore, in the receiver operating characteristic curve analysis, the area under the curve of the MPV and PDW to predict severe ED was 0.818 and 0.848 respectively. Our study establishes a dose-dependent association between the PDW and ED. Therefore, the PDW can serve as a potential marker for predicting the severity of ED.
  • Conventional slow freezing cryopreserves mouflon spermatozoa better than vitrification
    This work examines the effectiveness of a TCG (Tris, citric acid, glucose, 6% egg yolk and 5% glycerol) and a TEST (TES, Tris, glucose, 6% egg yolk and 5% glycerol) sperm extender in the freezing of mouflon spermatozoa at slow cooling rates, using different pre-freezing equilibration times (2–3 hr). It also examines the tolerance of mouflon spermatozoa to different concentrations of cryoprotectants (5, 10, 20% glycerol; 5%, 10%, 20% dimethyl sulfoxide; 6% polyvinylpyrrolidone) and/or sucrose (100, 300, 500 mm). The highest quality (p < .01) thawed spermatozoa were obtained when using the TEST extender and an equilibration time of 3 hr. Sperm motility and membrane integrity were strongly reduced when using rapid freezing rates (60–85°C min−1), independent of the concentration of cryoprotectants. The lowest sucrose concentration (100 mm) provided the highest (p < .05) percentage of motile spermatozoa and live spermatozoa with an intact acrosome. Vitrified–warmed sperm variables were at their best when the spermatozoa was diluted in TCG–6% egg yolk + 100 mm sucrose and warmed at 60°C. Slow warming at 37°C strongly reduced (p < .05) sperm motility and viability. However, sperm vitrification returned lower fertility, sperm motility and sperm viability values than conventional sperm freezing.
  • Association between personal basic information, sleep quality, mental disorders and erectile function: a cross-sectional study among 334 Chinese outpatients
    Male erectile dysfunction (ED) may cause anxiety and depression, while mental disorders and sleep disturbances may also be closely related to ED. However, the exact nature of their relationship remains unclear, and whether personal basic background data affect erectile function is unknown. We conducted a cross-sectional study among Chinese outpatients with ED from January 2012 to December 2014. All the men answered a questionnaire collecting information about mental health status, sleep disturbances and personal data, underwent a physical examination and had a blood sample drawn. Sleep disturbances were assessed on the basis of a 19-item version of the Pittsburgh Sleep Quality Index, which includes questions on sleep patterns during the past month. Among the 462 patients, 128 patients with alcohol abuse, diabetes, hypertension, hyperlipidaemia, psychiatric drugs, neurologic injury or abnormal hormones were excluded from the study; 86.27% and 68.66% of the patients suffered from anxiety and depression respectively. Sleep quality and anxiety symptoms significantly affected erectile function, whereas personal income and education level had no significant effects. Our study suggested that it is necessary to pay attention to the psychological status of patients with ED, especially anxiety disorder. Sleep quality may be an important factor affecting erectile function according to the personal data.
  • Effects of two-year testosterone replacement therapy on cognition, emotions and quality of life in young and middle-aged hypogonadal men
    The aim of the study was to examine the effects of two-year testosterone replacement therapy on cognitive functioning, emotional state and quality of life in young and middle-aged men with hypogonadotropic hypogonadism. Nineteen males diagnosed with hypogonadotropic hypogonadism participated in the study. Cognitive functions were assessed by Trail Making Test and Digit Span Test of Wechsler Adult Intelligence Scale. Emotional state was evaluated by Profile of Mood States. Quality of life was evaluated by WHO Brief Quality of Life Questionnaire. Changes after two-year testosterone replacement therapy were detected in Trail Making A (42.9 ± 22.3 vs. 36.2 ± 22.5, p = .050) and B (90.6 ± 55.3 vs. 65.6 ± 21.4, p = .025) tests, showing improvement in attention and visual scanning abilities, executive function and psychomotor speed, as well as in Digit Span Test forward score (5.4 ± 2.0 vs. 6.1 ± 2.6, p = .046), showing improvement in attention capacity and psychomotor speed. No significant differences were observed in emotional state and quality of life. In conclusion, beneficial effect in cognitive functioning (improved attention and visual scanning ability, executive function and psychomotor speed), but not in emotional state and quality of life, was observed in young and middle-aged hypogonadal men after two-year testosterone replacement therapy.


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