This study analysed the relationship of plasma testosterone with β-cell secretion, insulin sensitivity and other pituitary-target gland hormones in normoglycaemic adult men. The sample frame was the ‘Offspring of individuals with diabetes study’ database. A total of 358 offspring of individuals with type-2 diabetes (T2DM) and 287 individuals without known family history of T2DM were recruited for the study. Normoglycaemic men aged ≥18 years (maximum 55) were selected for this analysis. All participants underwent 75 g oral glucose tolerance test (OGTT); blood samples were collected at 0, 30, 60 and 120 min for plasma insulin and C-peptide. Total testosterone, cortisol, adrenocorticotropic hormone, thyroid stimulating hormone and thyroxine (T4) were measured in the fasting sample. A total of 164 men (age 28 ± 7.7 years) were included in analysis. Testosterone correlated negatively with BMI, waist to hip ratio (WHR), area under curve (AUC) of C-peptide and insulin (during OGTT) and was positively correlated with insulin sensitivity (r ~ 0.4). Cortisol and T4 positively correlated (weak) with testosterone (r ~ 0.2). In multivariate analysis, AUC C-peptide, BMI, WHR (negatively) and cortisol (positively) were related to testosterone. Concluding, testosterone correlated negatively with BMI and β-cell secretion. There was a positive association of testosterone with insulin sensitivity, cortisol and T4.

The aim of the study is to investigate the methods of separation, culture and identification of Sprague Dawley (SD) rat corpus cavernosal vascular endothelial cells (CCECs). Cavernosal tissues were isolated from male SD rats. Enzymatic digestion was applied to separate CCECs. Purified cells were obtained using immunomagnetic beads and flow cytometric cell sorting and subcultured in EMG-2 medium. The growth curve of CCECs was measured by the tetrazolium salt 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The cells were identified by von Willebrand factor (vWF) using immunofluorescence, and the positive percentage of vWF expression was detected by flow cytometry. The monomorphic cobblestone-like cells were observed by microscopy. High purification was obtained using immunomagnetic beads. After 2 days of incubation, cells entered the logarithmic growth phase and reached a plateau on the fifth day. The vWF expression in cytoplasm was positive. The purity of cells was 95.8%, which was tested by flow cytometry. SD rat CCECs can be separated and cultured successfully by the method of enzymatic digestion, immunomagnetic beads and flow cytometric cell sorting.

Detection of activated caspases of spermatozoa could be helpful to evaluate male infertility. Although western blot is validated as a highly specific method to detect the proteins extracted from cells, the ability of this technique to detect activated sperm caspases in human semen may be limited. Indeed, round cells, which potentially contain some activated caspases, may be present in semen and interfere with the detection of activated sperm caspases. Moreover, it is necessary to evaluate the minimum amount of spermatozoa necessary to optimise the detection of activated caspases in semen samples. Our results showed that interference due to round cells contained in semen with activated caspase-3 requires separation of spermatozoa by density migration. This sperm preparation selects a mature sperm population that does not reflect the whole sperm population, and in infertile men with oligoasthenoteratozoospermia, the amount of spermatozoa thus selected is usually low. Moreover, the western blot technique's low detection sensitivity and the low level of caspase enzyme activity in human spermatozoa for activated caspase-3, -8 and -9 mean that large quantities of spermatozoa are needed to detect the expression of the activated caspases. These limitations prevent this method being used for routine analysis in clinical practice.

Seminal plasma is a potential source of biomarkers for many disorders of the male reproductive system including male infertility. The identification and characterisation of differentially expressed proteins in seminal plasma of man with normal and impaired spermatogenesis can help in the elucidation of the molecular basis of male infertility. We compared the protein expression profiles of seminal plasma from four different groups of men as follows: normozoospermic, asthenozoospermic, oligozoospermic and azoospermic groups, using two-dimensional differential gel electrophoresis (2-D DIGE). We found eight proteins with statistically significant increased expression in azoospermia compared with at least one of the other studied groups. The differentially expressed spots were fibronectin, prostatic acid phosphatase (PAP), proteasome subunit alpha type-3, beta-2-microglobulin, galectin-3-binding protein, prolactin-inducible protein and cytosolic nonspecific dipeptidase. Notably, PAP was increased in patients with azoospermia compared with that of all other groups. We have observed no statistically significant differences in protein expression between three of the groups: normozoospermic, oligozoospermic and asthenozoospermic. We suggest that the identified panel of proteins in our study especially PAP have a strong potential to be used as azoospermia markers. However, further investigations will be necessary to validate these markers in samples of larger and independent patient cohorts and to clarify their role in the pathogenesis of male infertility.

Forecast of success with testicular sperm extraction and intracytoplasmic sperm injection (ICSI) remains unknown, as predictive factors have rarely been studied. We evaluated the association among possible predictive factors and a successful biopsy and clinical pregnancy. A consecutive sample of men submitted to a testicular open biopsy in S. João Hospital was used. Patient's age, medical history, testicular volume, spermogram, genetic testing, endocrinologic results, biopsy results and clinical pregnancy information were collected. From the 113 men included, it was possible to retrieve spermatozoa in 79.6% of the cases, which resulted in 58 fertilisations and 22 clinical pregnancies. Retrieving viable spermatozoa on biopsy was associated with the identification of spermatozoa in the spermogram (100.0% versus 74.4%; P = 0.010), diseases causing obstructive infertility (100.0% versus 79.2%; P = 0.036) and no genetic causes detected (82.4% versus 54.5%; P = 0.030). Successful clinical pregnancy was only associated with lower female partner age (31.7 versus 36.0 year; P = 0.001) but not the quality of the spermatozoa or the time until the reproduction cycle. Identification of spermatozoa in the spermogram, diseases causing obstructive infertility and lack of genetic causes for infertility were associated with higher probability of viable spermatozoa retrieval but the female partner age remained the principal determinant of a successful pregnancy.

The damaging effect of hyperlipidaemia on testicular structure was determined, and the influence of melatonin was evaluated in testicular damage related to hyperlipidaemia. Hyperlipidaemia was induced in ApoE-knockout C57BL/6J male mice fed with high-fat diet alone (group A), or with high-fat diet and melatonin (group B). Six ApoE wild-type C57BL/6J male mice were fed with normal diet, served as controls. At the end of the experimental period, ultrastructural observations showed dramatically histopathological alterations in testicular tissues of group A. The basement membranes of seminiferous tubules were partially thickened and wavy-like in testes of mice with hyperlipidaemia, and vacuolar degeneration of mitochondria and dilation of endoplasmic reticulum were identified as well as the number of mitochondria and lipid droplets decreased significantly in Leydig cells and Sertoli cells. Electrondense deposits were observed in cytoplasms of germ cells. The testicular histostructure in group B treated with melatonin was similar to that of control. Apoptosis was determined by terminal-deoxynucleotidyl-transferase-mediated dUTP nick end labelling. Apoptotic germ cells were significantly more numerous in group A than in group B and controls. The results suggest that melatonin may be potential to attenuate testicular damage by improving histopathological changes and reducing germ cell apoptosis in hyperlipidaemic mice.

The aim of the present study was to investigate the ameliorative effect of curcumin (CMN) against acute cadmium chloride (CdCl₂) toxicity on male reproductive system in rats. CdCl₂ is known to be a heavy metal and potential environmental pollutant. For this purpose, 28 rats were equally divided into four groups; the first group was kept as control and given distilled water and corn oil as carrier. In second and third groups, CdCl₂ and CMN were administered at the dose with 1 mg kg⁻¹ day⁻¹ and 100 mg kg⁻¹ for 3 days respectively. CdCl₂ and CMN were given together at the same doses in the fourth group. It was determined that acute CdCl₂ exposure caused a significant reproductive damage via increased oxidative stress (increased TBARS levels and decreased SOD, CAT, GPx and GSH levels), histological alterations (necrosis, oedema etc.) and spermatological damage (decreased sperm motility and sperm concentration and increased abnormal sperm rate) in male rats. However, CMN treatment partially reversed these toxic effects of CdCl₂ on the reproductive system. In conclusion, our results show that acute exposure of CdCl₂ may lead to infertility, and CMN could prevent and reverse hazardous effects of CdCl₂ to some degree. Thus, CMN may be useful for the prevention of CdCl₂-induced reproductive damage.

Information on male potency in testicular cancer (TC) patients treated with chemotherapy is insufficient. We aimed to assess the levels of depression and anxiety symptoms, sexual function and gonodotrophins. Participants (n = 27) were identified and recruited from the genitourinary services of two medical centres, one in Inonu University and the other in the Firat University. All patients are TC patients treated with chemotherapy after unilateral orchiectomy. Participants completed follow-up assessments after the completion of the chemotherapy regimen. Serum luteinising hormone, follicle-stimulating hormone and testosterone levels were determined after blood samples had been taken in the morning after an overnight fast. International Index of Erectile Function (IIEF-15) was also used to evaluate erectile dysfunction (ED) score. Beck Depression and Beck Anxiety Scale were used to assess psychological symptoms. The findings indicated that men treated with chemotherapy had significantly different IIEF-15 and Beck Anxiety scores compared with men who did not receive chemotherapy. But no statistically significant difference was determined in the serum gonodotrophin levels and depression score between the two groups. It is concluded that patients with TC undergoing chemotherapy have greater risk than normal men for ED, independently of the gonodotrophins level.

Our aim was (i) to investigate the hypothalamo–hypophyseal–gonadal axis in hyperthyroid Indian males, (ii) to rule out the modulatory role of adrenal steroids on it and (iii) to determine if the simultaneous rise in oestradiol and luteinising hormone (LH) in hyperthyroid males is due to a positive feedback action of oestradiol on pituitary LH release. Age- and BMI-matched men were divided into two groups, I, euthyroid subjects (n = 17) and II, hyperthyroid patients (n = 12) on the basis of their thyroid hormone levels. Serum levels of thyroid-stimulating hormone, triiodothyronine, thyroxine, LH, follicle-stimulating hormone (FSH), prolactin, E₂, T, P₄, sex hormone binding globulin and dehydroepiandrosterone sulphate (DHEAS) were assayed. Mean levels of T and E₂ were approximately two times higher in group II in comparison with group I. DHEAS levels were similar in both groups ruling out any adrenal involvement. Mean serum LH level was 2.6 folds higher in group II in comparison with group I. Mean serum levels of FSH were higher in group II, it was marginally nonsignificant. On the basis of these and previous observations, we hypothesise that endocrinological dimorphism in human male and female is not rigid; a sustained rise in serum oestradiol probably induces a positive feedback action on pituitary leading to elevated gonadotrophin levels.

In sows, the oviductal sperm-binding glycoprotein (SBG), which binds to the periacrosomal region of boar sperm, has been shown to be involved in sperm selection. In this work, we isolated porcine sperm proteins that interact with SBG. One of them is identified as a homologue of human S100A7 (psoriasin). Anti-human S100A7 antibodies show that this homologous protein localises to the head of sperm. The isolation of a homologue of S100A7 based on affinity to SBG and its localisation at the head of sperm leads us to suggest that S100A7's homologous protein may be involved in the negative selection of sperm by SBG in pigs. Human S100A7 shows antibacterial properties, particularly over Escherichia coli, a species that has demonstrated deleterious effects on human sperm. We searched for S100A7 in human sperm and found that it is present and localises at the acrosomal region. Thus, we report the presence of S100A7 in human sperm and of a homologous protein in pig, with similar localisations. In humans, an antimicrobial role seems likely for psoriasin; in porcine sperm the studied protein binds to SBG suggesting a function in sperm selection, but an antimicrobial function cannot be ruled out.

Latrodectus mactans’ aracnotoxin (Atx) induces changes in sperm function that could be used as a co-adjuvant in male contraceptive barrier methods. This effect includes the suppression of intracellular reactive oxygen species (ROS), an event necessary for capacitation, chemotaxis and acrosome reaction (AR). The sperm that are not trapped by the barrier method can reach the oviduct before fertilisation and be exposed to the secretions of the oviducts. This study evaluated the effect of bovine tubal explants (TU) and conditioned media (CM) from the ampullar and isthmal regions on spermatozoa exposed to Atx. Thawed bovine sperm were incubated with Atx, TU and CM from the ampullar and isthmal regions for 4 h and then DNA integrity, intracellular ROS and lysophosphatidylcholine-induced AR were determined. Spermatozoa exposed to Atx and co-incubated with TU and CM for 4 h produced an increase in sperm DNA damage, a decrease in ROS production and a decrease in %AR, compared with the control. A similar result was obtained from the co-incubation of spermatozoa with Atx. In conclusion, the effect of Atx is not modified by tubal cells or their secretions and this opens the door to future studies to evaluate the application of synthetic peptides obtained from Atx as a co-adjuvant of contraceptive barrier methods.

Intracytoplasmic sperm injection (ICSI) can be considered the most ‘revolutionary’ in vitro insemination technique because it has efficiently allowed the treatment of male factor infertility. Although ICSI has been successfully and safely applied worldwide for almost 20 years, currently, we have no real knowledge regarding the hypothetical long-term side effects on ICSI adults, given the increased likelihood of spermatozoa with defective nuclear content fertilising the oocytes. The aim of this review article is to investigate the most recent advances of performing ICSI in the safest possible manner, thus, minimising the theoretical hazards of this procedure. To allow for substantiated recommendation which male gametes to choose for physiological ICSI an updated search was performed in Medline and Embase, from 1996 to June 2011. Recent technical advances allow operators to more or less simulate physiological conditions in the laboratory, reducing potential damage to the gametes. It seems possible to prevent fertilisation by DNA-damaged and chromosomal-unbalanced spermatozoa by selecting ICSI sperm by motility and/or maturation markers such as hyaluronic acid or other zona pellucida receptors. Furthermore, novel non-invasive imaging techniques can be valid tools for helping in the morphological selection of ICSI spermatozoa.

Microwave radiation resulted in degeneration, apoptosis or necrosis in germ cells at different stages. The molecular mechanisms by which microwaves induce spermatogenesis disorder have not been completely understood. Sertoli cells play crucial roles in mammalian spermatogenesis. Cytokines produced by Sertoli cells play pleiotropic roles in different conditions. At physiologically low concentration, TNFα, IL-1β and IL-6 behave as survival factors; while under pathological condition, these cytokines can induce apoptosis in testis. The effects of cytokines produced by microwave-radiated Sertoli cells on spermatogenesis are poorly understood. The purpose of this study was to determine the effect of cytokines produced by microwave-radiated Sertoli cells on the germ cells. We focused the effect of TNFα, IL-1β and IL-6 on the germ cells. The results showed that TNFα, IL-1β and IL-6 were increased in Sertoli cells after exposure to microwave radiation. These up-regulated cytokines can induce apoptosis and lipid peroxidation in the membrane of germ cells. In addition, germ cell apoptosis was associated with the up-regulation of Bax/Bcl-2 and caspase-3. These results suggest that cytokines produced by microwave-radiated Sertoli cells may disrupt spermatogenesis. Our data provided novel insight into the injury mechanism of germ cells induced by microwave radiation.

This study investigated the protective effects of melatonin (MLT) against doxorubicin (DXR)-induced testicular toxicity and oxidative stress in rats. DXR was given as a single intraperitoneal dose of 10 mg kg⁻¹ body weight to male rats at 1 h after MLT treatment on day 6 of the study. MLT at 15 mg kg⁻¹ body weight was administered daily by gavage for 5 days before DXR treatment followed by an additional dose for 5 days. Sperm analysis, histopathological examination and biochemical methods were used for this investigation. DXR caused a decrease in the weight of seminal vesicles, epididymal sperm count and motility and an increase in the incidence of histopathological changes of the testis. In addition, an increased malondialdehyde (MDA) concentration and decreased glutathione content, glutathione reductase (GR), glutathione-S-transferase (GST), superoxide dismutase (SOD) and catalase activities were observed. On the contrary, MLT treatment significantly ameliorated DXR-induced testicular toxicity in rats. Moreover, MDA concentration and GR, GST and SOD activities were not affected when MLT was administered in conjunction with DXR. These results indicate that MLT had a protective effect against DXR-induced testicular toxicity and that the protective effects of MLT may be due to both the inhibition of lipid peroxidation and increased antioxidant activity.

The study was aimed at evaluating a new diagnostic interview, arbitrarily named Structured Interview about male accessory gland infections (SI-MAGI), administered to three different groups of patients with MAGI according to ultrasound evaluation [prostatitis (P) or prostato-vesciculitis (PV) or prostato-vesciculo-epididymitis (PVE)]. We evaluated 110 consecutively selected infertile patients, with diagnosis of MAGI and underwent to transrectal and scrotal ultrasound (US) examination. After ultrasound evaluation, they were divided into three well-characterised, aged-matched groups: prostatitis (P; n = 45), prostato-vesciculitis (PV; n = 30) and prostato-vesciculo-epididymitis (PVE; n = 35). Then, all patients were submitted to active clinical history oriented by SI-MAGI questionnaire elucidated by the operator. The SI-MAGI was structured in four domains (urinary symptoms, ejaculatory pain or discomfort, sexual dysfunction and quality-of-life impact) for a total of 30 questions with four possible answers. Infertile patients of PVE and PV groups showed scores significantly higher than P group in all domains (anovaP < 0.005). PVE showed scores significantly higher than PV group in domain 2 and 3. Our study demonstrates the MAGI’s symptoms worsen with major extension of inflammation and ultrasound can evaluate the true extent of MAGI and symptoms can help to suggest the severity of MAGI.

Late-onset hypogonadism (LOH) or age-associated testosterone deficiency syndrome is defined as a clinical and biochemical syndrome associated with advancing age and characterised by symptoms and a deficiency in serum testosterone levels. This condition may result in significant detriment in the quality of life and adversely affect the function of multiple organ systems. It has been suggested that sex steroid hormones may play a causal role in the development of insulin resistance and type II diabetes. This comparative study was aimed at determining the prevalence of LOH in diabetic men with erectile dysfunction and investigating the effect of testosterone replacement therapy on erectile function and on glycaemic control.

Solid surface vitrificaition (SSV) is a cryoperservative method that has been used in the cryopreservation of oocytes, and embryos. Here, we report an application of the SSV in the cryopreservation of human spermatozoa. We compared the SSV with a standard freezing method in terms of sperm motility, morphology, vitality and DNA integrity. Sperm motility was determined by computer assisted semen analysis, morphology and vitality were determined by eosin-methylene blue staining, and DNA integrity was determined by a TUNEL assay. We found that while both cryopreservative methods produced spermatozoa with comparable vitality and motility, the SSV gave slightly, but significantly fewer sperm with DNA damage, and loose tail. We concluded that, a cryopreservation of human spermatozoa by SSV is feasible and provides a quick and practical way to preserve human spermatozoa with a comparable, if not better, quality of the preserved spermatozoa to the standard freezing method.

Lower urinary tract symptoms (LUTS) and erectile dysfunction (ED) are frequently encountered in ageing males. We compared the efficacy of alfuzosin 10 mg alone or in combination with sildenafil 50 mg in the treatment of LUTS due to benign prostatic hyperplasia. One hundred male patients older than 45 years were randomized to two groups containing 50 patients each; one group receiving alfuzosin 10 mg and the other group alfuzosin 10 mg combined with sildenafil 50 mg. International Prostate Symptom Score (IPSS), quality of life (QoL), maximum flow rate (Q_max), prostate volume and post-void residual urine were evaluated. The mean age was 60.2 ± 17.8. Mean data of evaluated parameters in both groups at the end of 3rd month compared with baseline values are given respective order as; 5.1 (26.8%) and 5.8 (28.2%) points decreases in IPSS; 1.6 (41.1%) and 1.8 (45%) points decreases in QoL; and 3.4 (29.6%) and 3.4 (33%) points increases in Q_max. The outcomes of our study cannot be interpreted in such a way to report that alpha blocker-PDE5 inhibitor combination has a better efficacy than alpha blocker treatment alone in patients with LUTS.

The aim was to determine the prevalence of compound genetic abnormalities in patients who are carriers of cystic fibrosis transmembrane regulator (CFTR) gene polymorphism and to compare our results with similar patients reported in the literature. One hundred and nine patients were identified to be carriers of CFTR gene polymorphism. Additional genetic testing for karyotype abnormalities or Y chromosome microdeletions (YMD) was performed. Three patients (2.75%) of 109 were identified to have compound genetic abnormalities. One patient had 5T/5T while the other had 6T/6T and the third had 9T/9T. The three patients had deletions of azoospermia factor regions (AZFa+b or AZFa+b+c). There were no karyotype abnormalities identified in our database. In the literature, four patients with compound CFTR mutations and YMD were identified, in three patients had karyotype abnormalities. In conclusion, compound genetic abnormalities in CFTR mutation patients can be a contributing factor when abnormal spermatogenesis is encountered.

The aim of this study was to assess scrotal thermography in diagnostics of varicocele and suggest potential diagnostic criteria. Twelve patients with clinically diagnosed varicocele were examined with scrotal infrared digital thermography, physical examination and ultrasound/doppler. The main outcome measure was evaluation of thermography diagnostic criteria for varicocele. Mean temperature at left pampiniform plexus was ≥34 °C in 83%, and at right pampiniform plexus in all cases was ≤34 °C. In 92% of patients, temperature at the left testicle was ≥32 °C, whereas at the right testicle it was >32 °C in 50% patients. Temperatures between left and right pampiniform plexus and between left and right testicle were significantly different with P < 0.0001 and P < 0.006 respectively. In all patients, temperature difference between pampiniform plexuses was ≥0.6 °C. In 92% of patients, temperature at left pampiniform plexus was equal or higher to thigh temperature with the mean temperature difference of 1.1 ± 1.1 °C. Temperature at right pampiniform plexus was colder than the thigh in 92% of patients. This study suggests diagnostic criteria of five thermographic signs to easily diagnose varicocele. Scrotal thermography presents feasible, short and low cost diagnostic method for varicocele. Further study on a larger number of patients and healthy participants is needed to evaluate sensitivity and specificity of this method.

Penile fracture is an uncommon and emergent urologic condition defined as traumatic rupture of the corpus cavernosum secondary to a blunt trauma of the erect penis. Tunica albuginea is thinned and stretched in the erect state, and a transverse tear in the corpus cavernosums may occur by a buckling force. However, penile dorsal vein tears may mimic penile fracture. Also, corporeal infection and purulent cavernositis are associated with trauma, cavernosography, priapism, intracavernosal injection therapy and penile prosthesis.

Varicocele has been identified as an important cause of male infertility where its influence on men’s reproductive capacity is due to its markedly diverse effects on the testicles. This study aimed to assess the value of ultrasonographic parameters of the spermatic veins at the inguinal and scrotal levels in varicocele diagnosis and post-operative evaluation. Forty-five infertile men associated with varicocele and 15 fertile men were subjected to history taking, genital examination and semen analysis. In addition, inguinal and scrotal ultrasonography was carried out pre-varicocelectomy and 3 months post-varicocelectomy. At both the scrotal or inguinal levels, the mean spermatic vein diameter demonstrated significant post-operative decrease compared with the pre-operative resting condition and on Valsalva’ manoeuvre. The mean diameters of the pampiniform plexus of veins also demonstrated significant decreases post-operatively compared with the pre-operative resting condition or on Valsalva’ manoeuvre. It is concluded that colour Doppler ultrasound is a reliable and noninvasive method that is useful not only for diagnosis but also for post-varicocele repair follow-up.

The study is to analyse the clinical data of the two groups of patients with different fertilisation outcomes after short-time insemination and probe into short-time insemination indications. A retrospective study including 558 cycles of short-time insemination in our centre during January 2009 to June 2010 which were divided into two groups by the method of fertilisation, 472 cycles by IVF and 86 cycles by Re-ICSI group according to fertilisation conditions to and compare the differences between the two groups in the indexes of average female age, average male age, duration of infertility, forward-moving sperm counts, abnormal sperm rate, proportion of primary infertility, etc. The differences between the two groups have statistical significance in duration of infertility, forward-moving sperm counts, abnormal sperm rate and proportion of primary infertility (5.02 ± 3.12 versus 5.86 ± 3.35; 83.06 ± 78.55 versus 54.02 ± 61.78; 83.56 ± 5.24% versus 86.31 ± 5.26%; 56.87% versus 74.42%; P < 0.05). The discrepancy between the two groups has no statistical significance in average female age and average male age (P > 0.05). Duration of infertility, forward-moving sperm counts, abnormal sperm rate and previous pregnancy history have great significance to predict fertilisation failure, so decisions should be made on the basis of the above indexes when evaluating whether a patient should conduct short-time insemination.

The objective of the present study was to investigate the effect of ethanolic extract of Butea superba (Roxb.) on erectile dysfunction in diabetic rats by the measurement of intracavernous pressure (ICP) and on cavernosal smooth muscle relaxation. Male Sprague–Dawley rats were induced to become diabetic by a single intravenous injection of Streptozotocin (55 mg kg⁻¹ body weight). The ethanolic extract at the concentration of 1, 10 and 100 mg kg⁻¹ BW was administered orally once a day to diabetic rats in each group for 4 weeks. Diabetic rats showed a significant decrease in both ICP and the relaxation of the cavernosal smooth muscle compared with the normal rats. The extract of B. superba significantly increased the ICP with the effective dose of 10 mg kg⁻¹ BW (61.00 ± 11.11 mmHg versus 39.61 ± 11.01 mmHg in the diabetic control group). Moreover, the B. superba-treated group also showed enhanced relaxation of the cavernosal smooth muscle with EC₅₀ of 1.17 mg ml⁻¹. These results suggest that the extract of B. superba enhanced penile erection in diabetic rats by increasing the ICP. This might be explained by the increased blood flow as a result of the relaxation of the cavernous smooth muscle.

The development of a sperm normally ends in the female reproductive track through an extra-testicular maturation process termed capacitation. Previous studies in several mammalian species have revealed the presence of glycoconjugates and their capacitation-correlated changes in localisation. In this study, we used Lens culinaris agglutinin (LCA) to characterise mannosylated glycoconjugates in mouse sperm and their changes in distribution associated with capacitation. In freshly isolated sperm, three biochemically distinct sperm populations (type I, II and III) with specific LCA labelling were noted. Prior to capacitation, glycosylated materials were detected in the head region and approximately 26%, 65% and 9% of sperm displayed each distribution pattern. Following capacitation, the distribution pattern of these mannosylated constituents was not altered; however, re-localisation of these components was observed because the percentage of sperm exhibiting type I and III pattern was significantly changed. Finally, thirteen mannosylated glycoproteins were detected in murine sperm and our lectin blot data confirm that their redistribution instead of proteolytic degradation is mainly responsible for cellular dynamics we observed in murine sperm head associated with capacitation. In conclusion, this study is the first to describe the biochemical identification of LCA-binding glycoproteins present in mammalian sperm, which undergo redistribution as they mature to acquire their full fertilising ability.

A strong positive correlation exists between teratozoospermia and reactive oxygen species production, which in turn has negative effects on their in vitro fertilisation outcome. Our aim of this study was to determine potential protective effects of α-tocopherol on teratozoospermia motility, viability, acrosome reaction and DNA integrity after 1-h in vitro incubation. Teratozoospermic semen samples were obtained from 15 volunteers aged between 20 and 30 years after 3–5 days of sexual abstinence. Samples were washed, centrifuged and incubated in 37 °C and 5% CO₂ until sperm swimmed-up. Spermatozoa were counted in the supernatant and divided into four groups, each contained 2 × 10⁶ sperm/ml⁻¹. Groups one to four were incubated for 1 h with Ham’s F-10 solution as control group, 10 μm A23187, 40 μmα-tocopherol and 10 μm A23187 + 40 μmα-tocopherol respectively. The results indicated that α-tocopherol has ability to enhance teratozoospermia viability and motility, while there were no ameliorative effects on acrosome reaction and DNA fragmentation. A23187 induced acrosome reaction in teratozoospermia and α-tocopherol significantly diminished this effect. In conclusion, although α-tocopherol could improve teratozoospermia motility and viability, its effects on DNA integrity and acrosome reaction ability as supplementation IVF culture media are not obvious.

Boar semen is extremely vulnerable to cold shock and it is also sensitive to peroxidation due to the high content of unsaturated fatty acids in the plasma membrane. Antioxidants exert a protective effect on the plasma membrane of frozen boar sperm. Fennel has been shown to contain antioxidant substances. Therefore, this study was performed to evaluate the effect of different concentrations of fennel added to the freezing extender on boar semen quality and lipid peroxidation after thawing. Semen collected from four boars was cryopreserved in lactose-egg-yolk extender or in the same extender with varying concentration of fennel essences: low (LF); medium (MF); high (HF). Analysis of data clearly indicated that higher concentrations of fennel produced significant improvement in total motility. Moreover, when fennel was included in the extender, a dose-dependent tendency to increase sperm viability was observed. In contrast, the addition of fennel had no effect on acrosome integrity or hypoosmotic swelling test (HOST) compared with the control. Malondialdehyde (MDA) formation decreased significantly in fennel groups, yielding similar results for MF and HF. Fennel seems a new antioxidant for use in sperm cryopreservation, but its particular effects on sperm physiology must be further studied, especially the causes of motility stimulation and its effect on lipoxidation.

The accumulated data suggest that inflammation can increase the level of reactive oxygen species (ROS), which contribute to impaired sperm function and male infertility. Therefore, we propose that inflammation-mediated production of ROS in male and female reproductive tracts hinder sperm fertilisation. To test this hypothesis, phorbol myristate acetate (PMA) with polymorphonuclear leucocytes (PMNs) was applied to generate endogenous ROS. We evaluated the time-dependent effects of ROS on human sperm motility, viability and mitochondrial membrane potential (MMP). The results showed that after treatment with PMA and PMNs, the motility of human spermatozoa significantly decreased to 50% on Day 1 and 15% on Day 4 compared with that of the, respectively, negative controls (P = 0.012). The viability of human spermatozoa decreased on Day 4 of PMA + PMNs treatment (P = 0.028). The MMP of human spermatozoa significantly decreased from Day 2 to Day 4 in the PMA + PMN group compared with that of the controls (P = 0.019). Taken together, the 4-day cultivation approach provided an accurate evaluation of sperm quality, especially sperm motility and MMP. Our findings indicated that endogenous inflammation increased ROS levels, which might induce sperm oxidative damage. Additionally, sperm motility might be one of the earliest and most sensitive indicators of this damage.

Oxytocin receptor (OTR) expressed in the rat penis and mediated the contractility of the corpus cavernosum smooth muscle both in vitro and in vivo, and OTR could maintain penile detumescence; however, the expression of OTR in diabetic rat penis remains unknown. In the present study, we investigated the expression of OTR in diabetic rat penis. The experimental rats were randomly divided into control group and STZ-diabetic rats group. The expressions of mRNA and protein were examined by real-time quantitative PCR, Western blotting and immunohistochemistry respectively. Erectile function was evaluated by measuring intracavernous pressure following electrostimulation of the cavernous nerves. mRNA and protein expression of OTR significantly increased in diabetic rats group compared with the control group. Erectile function of diabetic rats group significantly decreased compared with the control group. Our data showed that the expression of OTR significantly increased in diabetic rats group and OTR may involve in the development of diabetic erectile dysfunction.

Toxoplasmosis is one of the classical conditions known to have an adverse effect on female reproductive functions, but a few investigations into male reproductive parameters have been performed. This work was carried out to study the effects of Toxoplasma gondii on reproductive function in male rats. Male rats were infected with the RH strain of T. gondii tachyzoites, and following every 10 days from 10 to 70 postinfection (PI), the percentage of body weight to testis weight ratio as well as epididymal sperm parameters (number, motility, viability, and morphology rates), serum testosterone (ST), intratesticular testosterone (ITT), serum lactate dehydrogenase (SLDH), intratesticular lactate dehydrogenase and fructose in seminal vesicles and coagulating glands were measured. The results of the study showed sperm motility, viability and concentration rates were significantly decreased temporary after infection up to 70 days. Sperm abnormality was also increased during these days. In addition, temporary alteration in ST, ITT, SLDH, intratesticular LDH and fructose in seminal vesicle and coagulating gland was observed PI. These findings suggest that toxoplasmosis can cause impermanent impairment on the reproductive parameters of male rats.

Ejaculates from six pure Spanish stallions were split, and one subsample frozen in a commercial extender supplemented with the lipid soluble antioxidant butylated hydroxytoluene (BHT), while the other subsample served as control. After at least 4 weeks of storage, samples were thawed and post-thaw sperm quality analysed: sperm motility and kinematics using a CASA system, membrane and acrosome integrity and mitochondrial membrane potential using flow cytometry. The outcome of cryopreservation varied significantly among stallions. However, the supplementation with 1 mm BHT had no significant effect on any of the sperm parameters evaluated post-thaw.

The infrequent presence of spermatozoa in cryptozoospermic men ejaculate is a limiting factor in the treatment of them. Sometimes, this consideration impels us to apply meticulous microscopic search in ejaculate or testicular sperm extraction (TESE) method. The aim of this study was to assess putative effectiveness of sperm origin, ejaculated or testicular, in cryptozoospermia treatment. In this context, were evaluated intracytoplasmic sperm injection (ICSI) outcomes in two parameters including fertilisation rate (2PN) and embryo quality, independently. We compared the outcome in two groups: patients who underwent ejaculate/ICSI and ones who underwent TESE/ICSI process. Nineteen ICSI cycles performed with testicular spermatozoa and the rest of cycles (n = 208) carried out with ejaculated spermatozoa. Result analysis showed similar fertilisation rate between testicular and ejaculated spermatozoa (respectively, 60% versus 68%, P ≥ 0.05). Also, on the other hand, embryo quality did not show significant differences between two groups, except grade A with low significance. With regard to almost equal performance of both methods in results and being invasive of TESE as surgical sperm retrieval method, the use of ejaculated sperm more than testicular sperm should be recommended in patients with cryptozoospermia whenever possible.

Pesticides can be toxic to desirable plants and animals, including humans. The aim of this study was to investigate the reproductive effects of low doses of pesticides on male offspring of exposed pregnant mice. Three groups of five female mice were treated daily by oral gavage with dimethoate (5 mg kg⁻¹ per day), deltamethrin (5 mg kg⁻¹ per day) and their mixture at 5 mg kg⁻¹ per day from day 3 to day 21 of pregnancy. Fertility, sexual behaviour and a number of reproductive endpoints, such as organ weights, sperm evaluations and testicular histology, were examined on four adult male offspring of exposed pregnant mice. When compared with control, a dose of deltamethrin 5 mg kg j⁻¹ causes a decrease in the absolute and relative weight of the testes of exposed mice and it affects their fertility by reducing the density, mobility and vitality of sperm and increasing the number of abnormal forms of these cells (P ≤ 0.01). The same results were obtained in mice exposed to a dose of 5 mg kg j⁻¹ combination of dimethoate and deltamethrin. This study demonstrated that deltamethrin and combination of dimethoate and deltamethrin caused a decrease in the absolute and relative weight of the testes, which affected the sperm parameters of male offspring of exposed mice to a low dose of these pesticides during pregnancy.

Chronic prostatitis (CP) is one of the most common male urogenital diseases and a significant public health problem in industrialised countries. It is associated with a low quality of life and significant expense. Given the poor results achieved with antibiotics, scientific interest has turned to the use of natural substances with a known activity on prostate function. The aim of our study was to evaluate the effect of a new dietary supplement containing lycopene, epigallocatechin gallate, ellagic acid, selenium and zinc on semen parameters and on leucocyte concentration in seminal fluid and expressed prostate secretion (EPS) in patients with CP without infection [National Institute of Health (NIH) Category IIIA], in comparison with a control group with the same condition who did not undergo any treatment during the study period. Our data showed a statistically significant reduction in inflammatory parameters (leucocytes in seminal fluid and EPS) and a statistically significant improvement in progressive sperm motility and sperm morphology in patients treated with the supplement in comparison with the untreated group. Improvements were also seen in the pain score of the NIH-Chronic Prostatitis Symptom Index (CPSI), confirming that the reduced inflammation also resulted in a reduction in pain.

Azoospermia, which is the absence of spermatozoa in the ejaculate, is not a rare cause of male infertility. Inducible nitric oxide synthase (iNOS) is a calcium-independent NOS, which is present in the testis and involved in spermatogenesis, and apoptosis of Sertoli and germ cells. Twenty idiopathic infertile men presenting nonobstructive azoospermia were enrolled in this study, and testicular sperm extraction procedures were performed. Tissue extracts were dissected, and the fluid samples were investigated to determine the presence of spermatozoa. Histologic evaluation of the spermatozoa-present samples revealed that seminiferous tubules were normal and were lined by Sertoli cells and spermatogenic cells. However, in the spermatozoa-absent samples, the diameter of the seminiferous tubules was small, and Sertoli-cell-only syndrome was determined in most of the tubules. iNOS expression was very weak in Sertoli cells, germ cells, and in Leydig cells in the spermatozoa-present group. In the spermatozoa-absent group, the immunostaining was very intense in Sertoli and Leydig cells. Electron microscopy findings were supported the histologic results. In conclusion, complete germ cell loss and intense expression of iNOS in the Sertoli and Leydig cells in the spermatozoa-absent groups of azoospermic human testis suggest an essential role of iNOS in spermatogenesis.

Varicocele is a common cause of male infertility and reports indicate that varicocelectomy has a beneficial effect on male fertility. The aims of this study were to evaluate and compare the total acrosin activity along with DNA integrity in semen samples obtained from 70 varicocele individuals with male factors infertility presenting grades II and III varicocele before and after the surgery and 30 fertile individuals without any clinical presentation of varicocele. Total acrosin activity, protamine deficiency, DNA fragmentation, and semen parameters including sperm concentration, motility and sperm morphology were assessed by spectrophotometery, CMA3 staining, terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay and WHO criteria, respectively. Total acrosin activity (P = 0.03), percentage CMA3 positivity (P = 0.00) and TUNEL-positive spermatozoa (P = 0.04) were higher in the varicocele individuals before the surgery compared with the fertile individuals; yet, all the aforementioned criteria decreased significantly after surgery (P < 0.05). The results of this study reveal that DNA fragmentation and protamine deficiency, as negative parameters in fertility, improve post-surgery; however, total acrosin activity as a positive parameter in fertility is higher in the varicocele individuals compared with fertile and decreases to a value close to the fertile control post-surgery. High levels of total acrosin activity in varicocele individuals need more research in future.

Mammalian testis functions at a temperature and oxygen tension (pO₂) lower than the core body. Hypoxia-inducible factor-1α (HIF-1α) mediates the adaptive responses to hypoxia such as production of angiogenic vascular endothelial growth factor (VEGF) in a variety of cells and tissues. VEGF production in Leydig cells is stimulated by luteinising hormone (LH)/cAMP. We have conducted experiments to find out whether HIF-1α is involved in LH/cAMP-induced secretion of VEGF by Leydig cell-derived MA-10 cells. Both cobalt chloride (CoCl₂), an inducer of hypoxia, and 8-Br-cAMP enhanced HIF-1α activity followed by an increase in VEGF secretion. However, there was no change in mRNA levels of HIF-1α. Inhibition of HIF-1α activity by cyclosporine A (CsA) inhibited a rise in VEGF production in response to CoCl₂ as well as 8-Br-cAMP. Inhibitors of protein kinase A (PKA), extracellular regulated kinase 1/2 (ERK1/2) and phosphatidyl inositol-3 kinase/Akt (PI3-K/Akt) inhibited the increase in VEGF levels in response to both CoCl₂ and 8-Br-cAMP. The data suggest that HIF-1α is a mediator of hypoxia- as well as 8-Br-cAMP-stimulated production of VEGF in MA-10 cells; both the stimuli act through a common signalling cascade.

Sperm morphology has been associated with in vitro as well as in vivo fertilisation. The study aimed to evaluate the possible relation between the percentage of spermatozoa with normal morphology and the following sperm functional assays: (i) zona-induced acrosome reaction (ZIAR); (ii) DNA integrity; (iii) chromatin condensation; (iv) sperm apoptosis; and (v) fertilisation rates. Regression analysis was employed to calculate the association between morphology and different functional tests. Normal sperm morphology correlated significantly with the percentages of live acrosome-reacted spermatozoa in the ZIAR (r = 0.518; P < 0.0001; n = 92), DNA integrity (r = −0.515; P = 0.0018; n = 34), CMA₃-positive spermatozoa (r = −0.745; P < 0.0001; n = 92), sperm apoptosis (r = −0.395; P = 0.0206; n = 34) and necrosis (r = −0.545; P = 0.0009; n = 34). Negative correlations existed between for the acrosome reaction, and DNA integrity, while negative associations were recorded with the percentages of CMA₃-positive spermatozoa, apoptotic and necrotic spermatozoa. Sperm morphology is related to sperm dysfunction such as poor chromatin condensation, acrosome reaction and DNA integrity. Negative and significant correlations existed between normal sperm morphology and chromatin condensation, the percentage of spermatozoa with abnormal DNA and spermatozoa with apoptotic activity. The authors do not regard sperm morphology as the only test for the diagnosis of male fertility, but sperm morphology can serve as a valuable indicator of underlying dysfunction.

With increasing evidence for faulty paternal contribution to reproduction, there has been a steady increase in studies highlighting an association between sperm DNA damage, failed/delayed fertilisation and aberrant embryo development. Owing to prevailing ambiguity, the aims of the study were to analyse the genetic integrity of the male gamete and then to understand its association with age, standard semen parameters, lifestyle and occupational factors. The study included 504 subjects, attending university infertility clinic for fertility evaluation and treatment. Semen characteristics were analysed by standard criteria; terminal deoxynucelotidyl transferase–mediated nick end-labelling assay was employed for DNA damage assessment. The average incidence of sperm DNA damage in patients with normozoospermic semen parameters was <10%. Patients with oligozoospermia, severe oligozoospermia, oligoasthenoteratospermia, asthenoteratozoospermia and necrozoospermia had significantly higher level of sperm DNA damage (P < 0.001). Patients above 40 years of age had significantly high levels of DNA damage (P < 0.001) compared with their counterparts. Patients with varicocele and a history of alcohol consumption had higher incidence of spermatozoa with DNA damage (P < 0.01). Poor sperm characteristics in the ejaculate are associated with increased sperm DNA damage. Age-related increase in sperm DNA damage and association of the same with varicocele and alcohol consumption are also demonstrated.

Gynaecomastia has been associated with a large variety of drugs in the literature. However, a causal relation of the incidence of gynaecomastia to a certain drug should be considered only if sufficient and significant evidence can be obtained from the studies published. In this review, studies quoted in Medline were evaluated according to the Scottish Intercollegiate Guidelines Network (SIGN) grading system for clinical studies. Reports on 92 drugs were found in Medline in combination with gynaecomastia. An imbalance of the oestrogen/androgen ratio causes gynaecomastia. Also, prolactin has gynaecomastia-inducing properties. In 14 of the drugs quoted, the studies lead to a level of recommendation ‘A’. All these drugs have been designed to interfere with the production and action of sexual hormones or of prolactin. In 25 of the drugs, the level of recommendation was ‘B’. Besides those drugs in this group that have been designed for interference with the metabolism of steroid hormones or of prolactin, in drugs for acid-related disorders, diuretics, antiretroviral drugs, antimycotics, psychoanaleptics, alcohol gynaecomastia was described as an unexpected adverse effect. Studies on the association of drugs and gynaecomastia do not share a generally accepted definition of gynaecomastia; in this way, the informational value is limited.

Premature ejaculation (PE) is by far the most common male sexual complaint, with millions of men affected all over the world. It is estimated that up to 20–30% of all men may be suffering from various forms of PE. A variety of filler materials are widely used nowadays for soft tissue augmentation. The appropriate filler can restore symmetry, volume and create a smooth skin surface The aim of this pilot study was to evaluate the therapeutic effect of hyaluronic acid gel injection in patients with PE. Sixty men with self-reported PE who were referred to our outpatient andrology clinic (between January 2007 and January 2008) were included in this study. Participants were randomly assigned using random sampling numbers into two distinct groups. Group A (n = 30) received a single injection of 2 ml of hyaluronic acid gel (Hyalift 3.5% micronised hyaluronic acid) using the previously described fan technique. Group B (n = 30) received a single injection of 2 ml of hyaluronic acid gel using the multiple puncture technique. Twenty-three patients (46.9%) received injection by the fan technique, while 26 patients (53.1%) received it through the multiple-point technique. The mean intravaginal ejaculation latency time (IELT) increased significantly from 2.12 ± 1.16 to 7.71 ± 7.86 min, after 1 month of injection and then dropped to 5.32 ± 3.52 min, but still remaining significantly higher than the baseline values. Results from our present pilot study demonstrated the usefulness of the application of hyaluronic acid dermal fillers in the treatment of PE; however, further investigations in large cohorts with longer follow-up are definitely needed to obtain more consistent results.

In the acrosome reaction, the spermatozoon plasma membrane fuses with the outer acrosomal membrane, resulting in the release of the acrosomal content. Several compounds, such as sex steroids, are known to modulate the acrosomal exocytosis. Testosterone regulates various functions in male reproductive physiology; however, little is known about the relationship between testosterone and the acrosome reaction. Thus, our objective was to study the effect of testosterone on the acrosome reaction of human spermatozoa. To evaluate the acrosomal exocytosis, spermatozoa were incubated with testosterone (0.2, 2.0 and 20 nmol l⁻¹), progesterone and control medium for 60, 120, 240 and 1440 min. The acrosome reaction was assessed by staining with Hoechst 33258 and fluorescein isothiocyanate–conjugated P. sativum agglutinin lectin. In general, spermatozoa incubated with progesterone had the highest percentage of acrosomal exocytosis. The percentage of acrosome reaction obtained in the three treatments with testosterone differed from that observed for progesterone at 120, 240 and 1440 min (24 h). Additionally, significant differences were found between testosterone (2.0 and 20 nmol l⁻¹) and progesterone after 60 min. Differences between control and the three testosterone treatments studied were obtained only at 1440 min. In general terms, these results show that testosterone exerts no inductor effects on the acrosome reaction of human spermatozoa.

We aimed to investigate the long-term effects of microsurgical varicocelectomy on pain improvement and sperm parameters in patients with varicocele-related pain. A total of 72 patients who had undergone microsurgical subinguinal varicocelectomy because of varicocele with scrotal pain between 2004 and 2009 were included in the study. The patients were grouped according to the presence or absence of pain following surgical treatment. The patients expressing a decrease in frequency and severity of pain or minimal fullness sense were included in the pain-positive group. The mean follow-up period was 54.4 ± 18.06 months. There was left-sided varicocele in 80.3% of patients, while 19.7% had bilateral varicocele. In our study, complete success rate (Visual Analog Scale: 0–1) was found to be 79.2% (57/72), while failure (complete and partial failure) rate was 20.8% (15/72), and a significant difference was found (P < 0.05). Microsurgical subinguinal varicocelectomy is a reliable approach for clinically varicocele patients with scrotal pain complaints. Regardless of the type of pain, varicocelectomy significantly decreases pain. The success of the treatment is not related to the degree of varicocele.

To improve the boar sperm cryopreservation process, the influence of the sugar (lactose, trehalose) source and the cryoprotectant [glycerol, dimethylformamide (DMF)] on the success of freezing was investigated. Sperm samples were frozen in one of six extenders: lactose plus 3% glycerol (LG); lactose plus 1.5% glycerol and 1.5% DMF (LGD); lactose plus 3% DMF (LD); trehalose plus 3% glycerol (TG); trehalose plus 1.5% glycerol and 1.5% DMF (TGD); trehalose plus 3% DMF (TD). Effects on motility, viability, acrosome integrity and hypoosmotic test (HOST) were measured. The results showed that extender containing 3% glycerol retained the highest motility percentages. In regard to viability and acrosome integrity, all extenders yielded similar rates except for the decreasing values of TD. Endosmosis was diminished in TD and LD at 2 h (P = 0.0018), as compared with the others. The results of the study demonstrated that the use of DMF as a cryoprotectant adversely affected boar sperm quality after cryopreservation.

Testicular lymphoma represents the commonest testicular malignancy in men older than 50 years. MR imaging of the scrotum is an efficient supplemental diagnostic tool in the evaluation of scrotal diseases. We report two cases of primary diffuse large B-cell testicular lymphoma, presented in men over the age of 50 years. MR imaging revealed the presence of a hypointense intratesticular mass lesion on T2-weighted images, strongly and heterogeneously enhancing after gadolinium administration.

Polyorchidism is defined as the presence of more than 2 histologically proven testes. We report the case of a 20-year-old man with polyorchidism, presenting with right scrotal pain and right scrotal ovoid mass. Scrotal examination revealed two ovoid, mobile lumps with testicular sensation in the right side of the scrotum. Scrotal colour Doppler ultrasonography and magnetic resonance imaging confirmed the presence of double testes with common epididymis and common vas. Microscopic varicocele ligation was performed, and then, two ipsilateral testes were sutured together. The testes were then returned to the scrotum with fixation.

Vitamin B₁₂ (cobalamin) deficiency results in atrophy of seminiferous tubules and aplasia of spermatozoa and spermatid. The transmembrane protein amnionless (AMN) directs endocytosis of cubilin with its ligand, contributing to intrinsic factor-vitamin B₁₂ absorption. To understand vitamin B₁₂ transport in testis, we analysed AMN expression in developing mouse testes and in Leydig cells and speculated the possible role of AMN in testis. In testes, Amn mRNA levels were low until 14 days post partum (pp) and markedly increased from puberty onwards. In the interstitium, Amn mRNA levels were low at 14 days pp and increased at puberty (28 days pp) together with 3-beta-hydroxysteroid dehydrogenase type 6 mRNA. Strong AMN immunoreactivity was observed in early spermatocytes from 7 days pp, suggesting that AMN participates in meiosis. In Leydig cells, AMN was not observed until 14 days pp but was strongly expressed after 28 days pp, suggesting a positive relationship between AMN expression and functional differentiation of adult Leydig cells. Together, AMN may participate in meiosis in early spermatocytes and in functional differentiation of adult Leydig cells through the mediation of vitamin B₁₂ transport in the mouse testes. This is the first report on AMN expression in the germ cells and soma of mammalian testes.

The application of nutraceuticals in the field of male sexual function –in particular for erectile dysfunction (ED) – remains relatively under explored. In a group of 54 unselected men (35–75 years), consecutively presenting to our ED clinic and naive to other ED treatments, we carried out a single-blind, one-arm study to evaluate the effects of a 3-month supplementation with propionyl-L-carnitine, L-arginine and niacin on their sexual performance. All patients had the short-international index of erectile function (IIEF) questionnaire, global assessment questions (GAQs) and routine laboratory testing, at baseline and 3 months afterward. 51 (92%) patients of 54 completed the entire study period. After 3 months of treatment, a small, but statistically significant improvement in total and single items of the IIEF was found (Δ = 5.7 ± 4.1 P < 0.01). Analyses on GAQs revealed that treatment improved erections in 40% of cases, with a partial response occurring in up to 77% of subjects enrolled. These preliminary findings indicate that the favourable cardiovascular effects of nutraceuticals might also reflect on male sexual function with possible implication in the treatment and prevention of ED. This study documents a considerable patient’s interest toward nutritional supplementation – as first-line or adjunctive treatment to PDE5 inhibitors – that goes beyond the measurable increment in penile rigidity.

In vitro incubation and centrifugation is known to decrease human sperm quality. In the human body, besides its antioxidant effects, l-carnitine (LC) facilitates the transport of activated fatty acids from the cytosol to the mitochondrial matrix. In this study, we investigated the effect of LC on human sperm motility, viability and DNA oxidation after incubation and centrifugation, following the sperm preparation protocols of assisted reproduction. Normozoospermic semen samples (n = 55) were analysed according to the World Health Organization (WHO) guidelines. LC concentrations that are not toxic to spermatozoa as determined by sperm motility and viability were standardised after 2 and 4 h of incubation at 37 °C. Semen samples to which the optimal LC concentrations were added were also centrifuged for 20 min at 300 g and analysed for sperm motility, viability and DNA oxidation. Sperm motility was improved at 0.5 mg ml⁻¹ LC after incubation and centrifugation with 5 × 10⁶ sperm ml⁻¹. Higher concentration of LC (50 mg ml⁻¹) significantly decreased sperm motility and viability. LC did not alter the baseline of sperm DNA oxidation during both incubation and centrifugation. In conclusion, LC may enhance sperm motility following incubation and centrifugation, while it might not affect sperm viability and DNA oxidation.

Our previous studies have shown that the cystic fibrosis transmembrane conductance regulator (CFTR) was important for capacitation and fertilisation in mouse, guinea pig and human spermatozoa. However, it is unclear whether CFTR is correlated with ovum fertilisation rate. The present study was to test the possible relationship between spermatozoa CFTR protein expression rate in healthy men and ovum fertilisation rate during in vitro fertilisation. Ninety-four couples for female factor infertility for IVF-ET treatments were retrospectively studied. All the patients were divided into three groups based on the fertilisation rate of ovum in vitro. It was performed to explore whether there were differences in sperm CFTR protein expression rate among the three groups and the relevance between CFTR protein expression rate and ovum fertilisation rate. Our study showed that there was no significant differences in sperm CFTR protein expression rate among the three groups (F = 0.614, P = 0.544), and the relevance between spermatozoa CFTR protein expression rate and ovum fertilisation rate was not significantly different (r = 0.013, P = 0.904). These results further suggest that CFTR protein expression rate in healthy men spermatozoa was not associated with ovum fertilisation rate and thus we cannot predict ovum fertilisation results by sperm CFTR protein expression rate.

To control growing world population, there is a need for male contraceptive methods that are comparable to female contraceptives, but due to lack of knowledge or investigation, no sufficient safe and effective contraceptives were developed till now. In the present investigation, the effect of 100% methanol extract of Opuntia dillenii phylloclade on reproduction in male rats was studied. A first group (I) received vehicle alone to serve as control. The second group (II) was further divided into treated and recovery groups, and the plant extract at 50 mg kg body weight⁻¹ was administered orally for 30 days. Biochemical, haematological and histopathological analyses were carried out to reveal the effects on reproductive organs in the male rats. The weights of reproductive organs were recorded. It was found that the number of fertile males, number of inseminated females, number of litters delivered and testosterone levels were reduced significantly. Epididymal sperm count and motility were also significantly decreased. Biochemical parameters support the antifertility activity of O. dillenii i.e. decreases in protein, glycogen content and elevation in cholesterol level. Testes and sperm morphology were altered significantly. Haematological parameters have not shown any significant changes. It is concluded that 100% methanol extract of O. dillenii possesses antifertility effects on male reproduction without change in general physiology.

The aim of this study was to investigate the effect of adding n-3 fatty acids (FA) and α-tocopherol (VE) to semen extender on freezing ability and FA composition of bull sperm. Semen was collected from 10 Iranian Holstein bulls and was pooled. In the first experiment, semen was divided into 12 groups including 4 levels of n-3 FA (0, 0.1, 1, 10 ng ml⁻¹) and 3 levels of VE (0, 0.1, 0.2 mmol). The treatment of 0.2 mmol VE and 10 ng ml⁻¹ n-3 FA had the best post-thawed sperm characteristics (P < 0.01). In the second experiment, lipid composition of the latest treatment and control (without FA and VE) was determined. Adding n-3 FA increased docosahexaenoic acid (DHA) percentage before freezing and after thawing. The ratio of n-3 to n-6 before freezing was higher (P < 0.05) in treated group than in control, and this ratio in the fresh sperm was greater than in the post-thawed sperm (P = 0.1). Results suggested that adding DHA accompanied with an antioxidant to an extender could improve cryosurvival of bull sperm via altering membrane lipid composition.

This case report demonstrates a successful pregnancy after ICSI combined with hypo-osmotic swelling test in a couple with Kartagener’s syndrome with complete immotile ejaculated spermatozoa. Our result suggests that even for complete immotile spermatozoa, the use of hypo-osmotic swelling test is a good alternative to identify viable spermatozoa. When associated with ICSI, it can be a valuable tool to get fertilisation and pregnancy.

The active transport of sperm through the female genital tract is a very important factor in the fertilisation process. There is conflicting evidence as to whether seminal plasma has a positive or a negative effect on reproductive performance regarding the sperm transport in the female genital tract. In this study, we investigated if the perfused swine uterus is an appropriate model to study the effect of human seminal plasma on uterine contractility. It was found that repeated application of human seminal plasma caused a significantly higher rise in pressure and frequency of contractions in perfused swine uteri compared to buffer [a 1.8-fold and 1.5-fold pressure rise (Δt1SP, P = 0.025; Δt2SP, P = 0.004)] after the first and second application of seminal plasma respectively, and even a 2-fold pressure rise after the third application with a statistical significance of P = 0.007. The pressure rise caused by the seminal plasma over the entire period of application was calculated using a mathematical programme and was represented by the integral of the pressure curve over time. This perfusion model as well as the use of pressure-time integrals is suitable for patho-physiological studies of the uterus.

Although histamine has been suggested to be involved in the control of male sexual function, including the induction of penile erection, its role in the human corpus cavernosum penis is still poorly understood. The aim of our study was to evaluate the course of histamine plasma levels through different stages of sexual arousal in the systemic and cavernous blood of healthy male subjects. Thirty four (34) healthy men were exposed to erotic stimuli to elicit penile erection. Blood was aspirated from the corpus cavernosum and a cubital vein during the penile conditions flaccidity, tumescence, rigidity and detumescence. Blood was also collected in the post-ejaculatory period. Plasma levels of histamine (ng ml⁻¹) were determined by means of a radioimmunoassay. Histamine slightly decreased in the cavernous blood when the penis became tumescent. During rigidity, histamine decreased further but remained unaltered in the phase of detumescence and after ejaculation. In the systemic circulation, no alterations were observed with the initiation or termination of penile erection, whereas a significant drop was registered following ejaculation. Results are not in favour of the hypothesis of an excitatory role of histamine in the control of penile erection. Nevertheless, the amine might mediate biological events during the post-ejaculatory period.

Adult polycystic kidney disease (APKD) is one of the most common inherited disorders affecting one in 800–1000 live births. Extra-renal manifestation of APKD is not uncommon. Cysts involving the male and female reproductive system have been reported. However, fertility is affected only in male subjects. Among cysts involving the reproductive system, seminal vesicle cysts have been reported to be the most common. The effect of seminal vesicle cysts on male fertility has been controversial. Current literature reports that majority of men remain fertile. However, uraemia and its implications on fertility and abnormal semen parameters in men with seminal vesicle cysts must be taken into consideration. We herein present two patients with APKD with contrasting semen parameters and also review the current literature.

Androgen insensitivity syndrome (AIS) is an X-linked disorder due to mutations of androgen receptor (AR) gene. Various AR mutations have been identified, and the characterisation of these mutations greatly facilitates our understanding of AR structure–function. In this study, we have analysed an AR missense mutation (N771H) identified in patients with AIS. Functional analysis of the mutant AR was performed by in vitro mutagenesis–cotransfection assays. Compared to the wild-type AR, the dose–response curve of dihydrotestosterone-induced transactivation activity in the mutant AR was greatly shifted to the right and significantly decreased. However, the maximal efficacy of transactivation activity in the mutant AR was similar to that of the wild type. Receptor binding assay indicated that the mutant AR had an approximately 2.5-fold lower binding affinity to dihydrotestosterone compared to the wild type. Western blot analysis showed that the size and the expression level of mutant AR in transfected cells were comparable to the wild type. These data underscore the importance of asparagine at amino acid position 771 of human AR in normal ligand binding and normal receptor function, and a mutation at this position results in androgen insensitivity in affected subjects.

This study aimed to investigate the expression of myocardin and serum response factor (SRF) in the cavernous tissue of diabetic rats. The experimental diabetes model was induced in 8-week-old male Sprague-Dawley rats (200–220 g) by a single administration of streptozotocin. Both the diabetes mellitus group (DM group, n = 20) and the control group (NDM group, n = 10) were injected with a low dose of apomorphine to allow for the measurement and comparison of the corresponding penile erections. Western blot and qRT-PCR were used to determine the protein and mRNA expression levels of myocardin and SRF. Erectile function was significantly decreased in the DM group compared with the control group (P < 0.001). The mRNA and protein expression levels of myocardin and SRF were reduced in the cavernous tissue of diabetic rats compared with the control group (P < 0.001). It is concluded that diabetes inhibits the mRNA and protein expression of both myocardin and SRF in the cavernous tissue. This could play a key role in the development of erectile dysfunction in diabetic rats.

The occurrence of bacteria in sperm samples intended for in vitro fertilisation can compromise the outcome of assisted reproductive techniques. Effective semen processing procedures should therefore be implemented to remove bacteria from semen. Unfortunately, technique failure does occur whereby bacteria can be found in processed sperm preparations. To improve the effectiveness of semen processing, a novel centrifuge tube insert was developed to facilitate the layering of density gradients and semen, and to prohibit the re-infection of purified sperm pellets. The purpose of this study was to: (i) determine the prevalence and type of bacteria present in semen of patients participating in the Unit’s assisted reproduction program and (ii) evaluate the effectiveness of density gradient centrifugation with the novel tube insert, for the elimination of bacteria and yeast from spiked human semen samples. A survey in 2007–2010 indicated that 50% of semen samples were found to have positive bacterial cultures. Semen processing by means of density gradient centrifugation with the novel tube insert eliminated significantly more in vitro derived (spiked) bacteria and yeast from semen compared to processing without the insert (P < 0.004). Therefore, it is highly recommended that the centrifuge tube insert, ProInsert™, be incorporated into assisted reproductive programs.

This study aimed at evaluating the impacts of sperm quality of six national sperm banks on pregnancy rates (PRs) of artificial insemination with donor sperm (AID) in China. A large retrospective analysis was performed on 1877 insemination cycles in 1209 women in a unique setting during a 3.5-year period. Global PRs of 22.1% per cycle and 34.2% per patient were achieved. The PRs of the six banks varied from 15.5% to 29.0% (P = 0.011). Significant differences were observed in the quality of donor semen provided by the six sperm banks. Moreover, in some banks, the poor sperm quality was related to the suboptimal PRs. However, in certain banks, high values of sperm parameters did not result in satisfactory PRs accordingly. These data demonstrated that variability of donor semen quality existed in the different banks. But, sperm parameters after thawing may not be detrimental factors affecting the success rate of AID treatment. Further studies are needed to seek potential molecular markers for predicting fertility potency of donor sperm.

Leydig cell tumours (LCTs) of the testis are rare. Their origin is still unknown. This case report describes a potential relationship between LCT and prolonged exposure to Finasteride.

Vardenafil hydrochloride (HCl) is a potent and selective phosphodiesterase type-5 (PDE-5) inhibitor that enhances nitric oxide (NO)-mediated relaxation of human corpus cavernosum and NO-induced rabbit penile erection, and enhances erectile function in patients. In the present study, the effect of vardenafil on nitric oxide synthase (NOS) and neuronal NOS expressions in the paraventricular nucleus (PVN) of rats without sexual stimulation was investigated using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and neuronal NOS (nNOS) immunohistochemistry and western blot analysis. The present results showed that NOS and nNOS expression in the PVN was increased by vardenafil treatment as the dose- and duration-dependently without sexual stimulation. The phosphodiesterase type-5 inhibitor, vardenafil, augmented NOS expression in the brain without sexual stimulation. The present study suggests that sexual behaviour can be directly modulated by neurotransmitters such as nitric oxide.

Male infertility is a heterogeneous disorder that contributes to the impairment of spermatogenesis. The purpose of this study was to assess whether tumour suppressor p53 gene (Tp53) polymorphism is associated with idiopathic male infertility in the Iranian population. The prevalence of G→C substitution at codon 72 in exon 4 was determined in 110 idiopathic infertile azoo-/oligospermic patients and 180 fertile healthy control men. PCR-restriction fragment polymorphism analysis was employed to determine the genotypes. PCR amplicons were subjected to restriction digestion with Bstu1 and separated by gel electrophoresis. The Arg/Arg genotype was found more frequently among men with idiopathic infertility (32.7%) than among controls (17.7%). No significant difference was observed between Pro/Pro genotype and Arg/Arg + Arg/Pro genotypes among men with idiopathic infertility and controls (P = 0.11; OR: 0.36; 95% CI: 0.36–1.10). In contrast, a significant difference was observed in the comparison of the Arg/Arg genotype and Arg/Pro + Pro/Pro genotypes among patients and controls (P = 0.004; OR: 2.25; 95% CI: 1.29–3.90). Allele frequency evaluation suggested a significantly higher incidence of the Arg allele among infertile men compared with controls (56% versus 44%; OR: 1.6; 95% CI: 0.92–2.80). In conclusion, arginine allele appears to be at greater risk of developing idiopathic infertility in Iranian men.

The mechanism of testicular toxicity of lead (Pb) and cadmium (Cd) is poorly understood. Previous studies focused on single metal-related changes in testicular toxicity. This study points towards the possible involvement of Pb- and Cd-induced oxidative stress in the suppression of steroidogenesis. The oxidative status of testis of adult male rats exposed to Pb acetate and cadmium acetate either alone or in combination at a dose of 0.025 mg kg⁻¹ body weight of metal intraperitoneally for 15 days was studied. Pb and Cd caused an increase in reactive oxygen species (ROS) by elevating testicular malondialdehydes (MDA) and decrease in activities of testicular antioxidant enzymes superoxide dismutase (SOD), catalase, glucose 6 phosphate dehydrogenase (G6PDH) and glutathione-S-transferase (GST) in mitochondrial and/or post-mitochondrial fraction. Activities of steroidogenic enzymes 3β and 17β-hydroxysteroid dehydrogenase also decreased significantly leading to altered testosterone production. Metal-exposed groups showed significantly decreased testicular and epididymal sperm count. Epididymal sperm motility and viability was also decreased on Pb and Cd exposure. Cd exposure showed more toxic effect than lead exposure, while combined exposure demonstrated least toxicity. In vitro experiments showed that vitamin C restores steroidogenic enzyme activities, suggesting that Pb- and Cd-induced ROS inhibits the testicular steroidogenesis.

This study investigated the effects of gestational and lactational exposure to lead and cadmium on testicular steroidogenesis, antioxidant system and male accessory gland functions in F1 generation rats to understand the biochemical mechanisms involved in endocrine disruptions. Pregnant rats were subcutaneously administered with 0.05 mg kg⁻¹ body wt day⁻¹ of sodium acetate (control), lead acetate, cadmium acetate and (lead acetate + cadmium acetate) throughout the gestational–lactational period, and all animals from each of the experimental groups were sacrificed by decapitation on post-natal day 56 for performing various biochemical assays. We observed significant reduction in the activities of testicular key steroidogenic enzymes and serum testosterone concentration along with significant depletion in cholesterol, ascorbic acid and reduced glutathione contents in all the metal-treated groups. Reductions in the activities of catalase and superoxide dismutase with concomitant increase in the levels of thiobarbituric acid reactive substance were observed in experimental groups. Both sperm contents and sperm motility patterns were significantly altered in all the metal-treated groups, suggesting the direct/indirect spermotoxic effects of lead and cadmium. The inhibitory effects of lead, cadmium and combined exposure on testicular steroidogenesis machinery, along with the male accessory gland functions, are indicative of multiple targets of lead and cadmium to disrupt male reproductive functions.

Adenosine deaminase (ADA; E.C.3.5.4.4) catalyses the deamination of adenosine to inosine. In the human reproductive system, the importance of enzymes that affect metabolism of adenosine, particularly adenosine deaminase, has been noticed. The purpose of this study was to determine the plasma activities of total adenosine deaminase (ADAT), and its isoenzymes, ADA1 and ADA2, in fertile and infertile men. Plasma activities of ADA and its isoenzymes were measured in 55 fertile men and 70 infertile men. There was a significant difference in the ADA1 and ADA2 activities between fertile and infertile individuals (P < 0.01). The activity of ADAT, ADA2 and ADA1 in infertile men was higher than that in fertile individuals. This alteration in ADA activity can lead to reduced adenosine levels, which may be involved in disturbing the fertility process.

The endogenous opioid system has been reported to have important functions in human reproduction. Practically all the components of this peptide system have been discovered in human sperm cells, but their functions in these cells are far from being well understood. In the present work, we report the effects of opioid agonism and antagonism on human sperm motility, a parameter which is crucially associated with male fertility. Morphine (10⁻⁷ m), a μ- opioid receptor agonist, decreased both the percentage of motile progressive sperm and three measured velocities without altering the linearity, straightness or vigour of sperm cells. This effect was reversed by naloxone. Higher doses of morphine did not have further effects on the measured parameters. The incubation of sperm cells with the δ-opioid receptor agonist D-penicillamine (2,5)-enkephalin did not affect sperm cell motility. However, naltrindole, a specific δ-receptor antagonist, reduced the linear and curvilinear velocities, as well as linearity, straightness and the amplitude of head displacement, and beat frequency. In summary, our results indicate that the endogenous opioid system may regulate opioid motility in vitro. These finding suggest that the endogenous opioid system could be useful as a biochemical tool for the diagnosis and treatment of male infertility.

The aim of this study was to evaluate whether the viscosity of semen in patients with male accessory gland infection is related to the extension of the inflammatory process to the various glands. To achieve this, viscosity was assessed by quantitative viscosimeter and the results were expressed in centipoise (cps). The study was conducted on 30 infertile patients with clinical evidence of male accessory gland infection and a mean age of 29.0 ± 4.0 years. Their semen viscosity was evaluated through quantitative viscometer. All patients showed an increase of viscosity evaluated according to WHO criteria, while this parameter was normal in all controls. Semen viscosity of patients with male accessory gland infection (28.6 ± 2.2 cps) was significantly (P < 0.05) higher than that in the controls (10.7 ± 0.6 cps). Significantly increasing values were observed in patients with involvement of multiple gland inflammation (prostatitis <prostato-vesiculitis < prostato-vesiculo-epididymitis). The viscometry allows us to identify a quantitative score more severely in more extended forms. It therefore represents a valuable diagnostic tool in clinical practice, which also allows better monitoring of treatment response of male accessory gland infection.

The aim of this study was to evaluate the level of malondialdehyde (MDA) (one of the final products of lipid peroxidation and well-known marker of oxidative stress) in semen of infertile men with type 2 diabetes and to investigate its relationship with their glycaemic control. Forty infertile men with type 2 diabetes were evaluated. The mean ages were 36.5 ± 8.0. Men with diabetes were divided into two groups. Group A (n = 20) with glycated haemoglobin >10% and group B (n = 20) with glycated haemoglobin <7%. A single sample was examined according to the criteria of the World Health Organization (WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction, 1999, Cambridge University Press). MDA was assessed using the thiobarbituric acid method. MDA concentration in semen of group A patients (0.95 ± 0.35 nmol ml⁻¹) was significantly higher than in group B patients (0.43 ± 0.13 nmol ml⁻¹) (P value < 0.05) and had negative relationship with sperm density (r = −.717; P value < 0.05), total sperm count (r = −.625; P value < 0.05), progressive motility (r = −.489; P value < 0.05) and normal forms (r = −.545; P value < 0.05). Based on these results, it could be concluded that increase in lipid peroxidation in men with diabetes with poor metabolic control was associated with low sperm quality.

Although the majority of patients with diabetes have disorders in sexual function, associations between diabetes mellitus and sperm function at the molecular level are largely unknown. As receptor for advanced glycation end products plays a key role in many diabetic complications, we hypothesised that it may be involved in sperm nuclear DNA fragmentation. RAGE levels were determined using ELISA and western blot analysis in sperm samples from 32 diabetic and 35 nondiabetic men. Sperm DNA fragmentation was assessed using TUNEL assay. Diabetic men had significantly higher mean levels of RAGE protein (P < 0.001) and DNA fragmentation (P < 0.001) in spermatozoa. Sperm RAGE was directly correlated to sperm DNA fragmentation in diabetic men (r = 0.81, P < 0.001). The high positive correlation between RAGE levels and nuclear DNA fragmentation in spermatozoa of diabetic men suggests a central role of RAGE in disturbances in sexual function of diabetic men.

The aim of this study was to investigate the effect of alpha lipoic acid (ALA, an universal antioxidant) on thinner-induced testicular toxicity regarding spermatological features, body and reproductive tract measures in rats. Adult male Sprague-Dawley rats were divided into five treatment groups, eight rats in each. Control group was treated with placebo. Group O was given only olive oil. The group L received only α-lipoic acid. Thinner + Lipoic Acid group received thinner + α-lipoic acid and group T received only thinner. Thinner alone administration caused significant decreases in body and some reproductive organ weights, sperm count, motility and sperm membrane integrity, and significant increases in seminal vesicle weight and abnormal sperm rates compared with the values in the control group. However, concomitant administration of thinner with α-lipoic acid provided significant improvements in sperm parameters compared with values in alone group T. In conclusion, the results of this study suggest that α-lipoic acid has a protective effect against thinner-induced reproductive dysfunction in male rats.

MAGI is the inflammation of the accessory male glands that notoriously exerts a negative influence on male fertility. The diagnosis is integrated by clinical, laboratory and ultrasound evaluation. In particular, the ultrasound criteria were published in 1999. The aim of this study was to analyse the sensitivity and specificity of additional diagnostic ultrasound criteria as well as of conventional criteria in a selected category of infertile patients with MAGI. To accomplish this, 100 patients with MAGI were evaluated by scrotal and transrectal ultrasound by three different operators. The control group consisted of 100 age-matched healthy men. Statistical analysis was performed to evaluate sensitivity, specificity, positive and negative predictive values, and ROC curve analysis. The results showed that additional ultrasound criteria had a diagnostic accuracy similar to traditional criteria. The threshold value of two criteria for each diagnostic category (traditional and additional criteria) obtained high values of sensitivity and specificity. In conclusion, this study confirms the validity of the ultrasound criteria of MAGI previously published; in addition, it suggests the clinical utility of other indicators in clinical practice with good diagnostic accuracy and finally it establishes a clear threshold ultrasonographic value for the diagnosis of MAGI.

The aim of the present study was to evaluate gene expression profile by microarray technology and validation by real-time PCR in paired samples of testicular biopsies (pre-surgery and post-surgery) in two patients with varicocele. The microarray analysis showed increased expression levels after surgery in 215 and 52 genes in patient 1 and 2, respectively, as well as decreased expression levels in 65 genes in patient 1 and 358 genes in patient 2. Real-time PCR confirmed the differential expression of the five selected genes: MT1M, PHLDA1 and INSL3 had decreased expression levels and CCIN and PRM2 increased expression levels compared with pre-surgery biopsies. In conclusion, both techniques showed decreased expression levels of genes involved in stress situations associated with varicocele and increased expression levels of genes involved in normal function of spermatogenesis.

The aim of this study was to evaluate the incidence of spermatic aneuploidies in men with severe teratozoospermia and to determine an eventual relation between aneuploidies and a specific morphology of spermatozoa. Fluorescence in situ hybridisation (FISH) using a probe cocktail containing the alpha satellite for the centromeric region of chromosome X, Y and 18 was performed on decondensed spermatozoa from fresh ejaculates of thirty patients with severe teratozoospermia (abnormal forms >80%) and 15 fertile men with normal semen profiles. The mean frequency of teratozoospermia in patients was 91 ± 6.99%. There was statistically a significantly increased frequency of 1818, XY, XX and YY disomies in sperm with severe teratozoospermia compared with normal sperm (1.24% versus 0.08%, 1.42% versus 0.31%, 1.13% versus 0.19% and 1.11% versus 0.24%, respectively, P < 0.001 in all comparisons). The rate of total diploidy was significantly increased in patients compared with controls (1.46% versus 0.16%, P < 0.001). There was a correlation between macrocephalic spermatozoa and diploidy (r = 0.37, P < 0.05). Our data add further evidence that patients with severe teratozoospermia have an increased sperm aneuploidy rate and that this is particularly high in macrocephalic spermatozoa; FISH analysis on sperm could help to improve risk assessment and reproductive counselling in these individuals who are frequently candidates for intracytoplasmic sperm injection (ICSI) as a treatment of their infertility, as the use of ICSI has created consequential debate concerning the genetic risk for the offspring.

The purpose of this prospective study was to evaluate Kruger strict morphology and conventional semen analysis in predicting cryosurvival and the progressive motility recovery rate of frozen spermatozoa. Our study included 56 semen samples with >10 million spermatozoa per ejaculate. The main outcome measures were conventional semen analysis, strict morphology analysis by the Kruger method, cryosurvival rate and post-thaw sperm motility. A significant reduction in sperm motility after cryopreservation was demonstrated. The freeze-thawing process caused a 66% reduction in rapid progressive motile spermatozoa, a 45% reduction in slow progressive motile spermatozoa and a 2% reduction in nonprogressive motile spermatozoa. The cryosurvival and progressive motility recovery rates were not correlated with parameters of conventional semen analysis, such as sperm concentration, motility, WHO morphology and total motile count, but the progressive motility recovery rate was significantly correlated with the percentage of spermatozoa exhibiting Kruger normal morphology (P = 0.028). The recovery rate of rapidly progressive motility was profoundly decreased compared with slow progressive motility following the frozen-thaw procedure of semen. Kruger strict morphology assessment was a better predictor of the progressive motility recovery rate following the freezing-thaw procedure than parameters of conventional semen analysis.

Studies have documented the presence of herpes viruses in semen. The aim of our study was to determine whether they persist in semen samples following two-density gradient centrifugation for IVF purposes. Semen samples were collected from 109 men seeking fertility evaluation, prior to IVF treatment. Routine semen analysis was performed according to WHO guidelines. Each sample was treated in a two-density gradient centrifugation using PureSperm (PS). Both untreated and treated samples were screened for the presence of herpes viruses, using PCR. Kruskal–Wallis, chi-square and binomial statistical tests were used; P ≤ 0.05 was considered statistically significant. No statistically significant associations were observed between semen parameters and viral presence. Viral DNA was detected in 54% of semen samples: HSV1/2 in 32 samples, EBV in 49, CMV in 47, HHV6 in 9, HHV7 in 4 and VZV in none. PS gradient failed to remove CMV in 89.36%, HSV1/2 in 59.38% and EBV in 22.45% of samples, while HHV6 and 7 were completely removed. Especially HSV1/2 and CMV seem to persist even following PS treatment. These observations indicate the possible risk of oocyte becoming infected during insemination, by IVF or intracytoplasmic sperm injection, with unknown sequelae. Further studies are required to determine whether any correlation exists between their presence, implantation rate and the outcome of pregnancy.

This study was conducted to evaluate the correlation between seminal plasma arginase activity and spermatological parameters in rams. In this study, five fertility-proven Awassi rams were used as material. Six ejaculates were collected from each ram by an artificial vagina. Spermatological parameters (semen volume, mass activity, sperm motility and concentration and abnormal sperm rate) were immediately determined in each ejaculate. For enzyme assay, the semen samples were centrifuged and stored at −20 °C for the analysis of arginase activity. The average seminal plasma arginase activity was 0.61 ± 0.20 U (mg protein)⁻¹. There was a positive correlation between arginase activity and semen volume (r = 0.412, P < 0.05), semen mass activity (r = 0.610, P < 0.01), sperm motility (r = 0.447, P < 0.05) and sperm concentration (r = 0.808, P < 0.01). However, there was a negative correlation between arginase activity and abnormal sperm rate (r = −0.424, P < 0.05). In conclusion, this study clearly suggests that there is a significant correlation between seminal plasma arginase activity and spermatological parameters. In light of these results, seminal plasma arginase activity may be a biochemical criterion for determining sperm quality besides classical semen analysis parameters in rams.

The present works examined an adverse effect of chlorpyrifos insecticide on testes and lipid peroxidation at low doses (5 mg–10 mg kg⁻¹ body weight) and the role of antioxidant enzymes systems at higher doses (20–30 mg kg⁻¹ body weight) in albino rats. At low doses, reduction in plasma levels of testosterone and FSH and LH hormones along with the significant shrinkage of seminiferous tubules and gametogenic changes in germ cells were noticed. But these changes were restored with the revival of serum testosterone, FSH and LH along with regression of testis at higher doses. Similarly, level of testicular lipid peroxidation was elevated, whereas levels of antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) and steroidogenic enzymes activities (Δ⁵, 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase) were reduced significantly at low doses. But, rat testes showed a significant decrease in lipid peroxidation and concomitant increase in antioxidant enzymes and steroidogenic enzymes activities at higher doses. Results showed that at higher doses of chlorpyrifos treatments, rat testes were shown to trigger their natural defence mechanism which became operative possibly through corrective measure of synthesis of antioxidant defence enzymes and steroidogenic enzymes and pituitary gonadotrophins hormone feedback mechanisms.

The aim of the study was to compare the semen characteristics and nuclear DNA fragmentation in spermatozoa of diabetic and nondiabetic men undergoing assisted reproduction and correlate them with pregnancy outcome. Semen characteristics and nuclear DNA fragmentation were analysed using computer-aided semen analysis system and sperm chromatin dispersion assay (SCD), respectively. Spermatozoa from diabetic patients showed significantly lower progressive (Type A) motility (14.64 ± 9.60 versus 17.99 ± 11.51, P < 0.02) and increased nuclear DNA fragmentation (37.05 ± 12.68 versus 21.03 ± 10.13, P < 0.001). Furthermore, a positive correlation was observed in diabetic patients in terms of blastocyst formation rate (38.13% versus 55.46%, P < 0.001), pregnancy rate (28.57% versus 46.34%, P < 0.001) and miscarriage rate (50.0% versus 24.56%, P < 0.001). The higher percentage of sperm DNA damage because of oxidative stress seen in diabetic patients may be responsible for the poor embryonic development and pregnancy outcome in these individuals.

There are few reports of pregnancy outcomes in couples with anejaculatory infertility secondary to multiple sclerosis and no longitudinal reports of semen quality in this population. We report our experience with one such case. The couple achieved two live births from spermatozoa obtained with electroejaculation: the first by intrauterine insemination and the second by in vitro fertilisation with intracytoplasmic sperm injection of donor oocytes. Linear regression analysis showed no progressive decline in semen parameters across 26 semen retrievals performed over 7.7 years. Years of disease do not appear to cause progressive decline in semen quality.

The aim of this study was to examine the possible relationship between sperm DNA integrity and chromatin packaging evaluated by cytochemical assays, traditional sperm parameters and recurrent spontaneous abortion (RSA) of unknown origin. In this cohort study, 40 couples with a history of RSA and 40 couples with proven fertility were considered as case and control groups respectively. The semen samples of all husbands were analysed for sperm parameters and also sperm chromatin and DNA integrity assessed using cytochemical tests including aniline blue (AB), chromomycin A3 (CMA3), toluidine blue (TB), acridine orange (AOT) and nuclear chromatin stability assay. Among different sperm parameters, only slow motility was significantly different between the two groups. In sperm chromatin evaluations, there were significant differences between the two groups in all of the tests. In addition, the majority of semen samples in RSA patients exhibited upper percentages of abnormal spermatozoa than the cut-off values regarding different cytochemical assays. Our study showed that in the cases of RSA, slow motility had a significant reduction in comparison with controls and also spermatozoa of men from RSA group had less chromatin condensation and poorer DNA integrity than spermatozoa that obtained from fertile men with no history of RSA.

In this study, it was aimed to determinate beneficial effects of protocatechuic acid (PCA) against reproductive toxicity caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), an environmental contaminant. For this purpose, 28 rats were equally divided into four groups (control, TCDD 2 μg kg⁻¹ per week, PCA 100 mg kg⁻¹ per day and TCDD + PCA group), and compounds were orally administered for 45 days. The results indicated that TCDD induced oxidative stress via an increase in thiobarbituric acid-reactive substances levels and a decrease in reduced glutathione, catalase, glutathione peroxidise and SOD levels in male rats. In contrast, PCA treatment prevented toxic effects of TCDD in terms of oxidative stress. Additionally, sperm motility, sperm concentration and serum testosterone levels significantly decreased, and pathologic testicular damage increased with TCDD exposure. However, these effects of TCDD on sperm characteristics, histopathological changes and hormone levels were reversed by PCA treatment. In conclusion, it was found that TCDD exposure induced reproductive toxicity (oxidative, hormonal, histopathological and spermatological alternations) in male rats and PCA treatment could prevent toxic effects of TCDD. Thus, PCA may be useful for the prevention and treatment of reproductive toxicity caused by TCDD.

Liquid nitrogen preservation in remote farms is a limitation. The goal of this study was to determine optimum temperature above freezing point for bovine spermatozoa preservation using bovine serum albumin (BSA) as a supplementation. Pooled semen sample from three ejaculates was subjected to various BSA concentration (1, 4, 8 and 12 mg ml⁻¹), before incubation in different above freezing point temperatures (4, 25 and 37 °C). Viability assessment was carried out against time from day 0 (fresh sample) until all spermatozoa become nonviable. Optimal condition for bovine spermatozoa storage was at 4 °C with 1 mg ml⁻¹ BSA for almost 7 days. BSA improved bovine spermatozoa viability declining rate to 44.28% at day 4 and 57.59% at day 7 compared to control, with 80.54% and 98.57% at day 4 and 7 respectively. Increase in BSA concentration did not improve sperm viability. Our results also confirmed that there was a strong negative correlation between media osmolarity and bovine spermatozoa survival rate with r = 0.885, P < 0.0001. Bovine serum albumin helps to improve survival rate of bovine spermatozoa stored above freezing point.

The use of ureteral stents is a common procedure in urology practice. We investigated whether the use of double-J stent had an effect on sexual functions or not. One hundred and seventy-seven cases were included in the study. Unilateral double-J stent was placed during ureteral stone treatment in one hundred and eight of these cases. Sixty-nine cases were included as the control group. ‘International Index of Erectile Function’ (IIEF) and ‘Female Sexual Function Index’ (FSFI) questionaries were assessed before and 4 weeks after the intervention in all patients. When total IIEF and subdomains of IIEF scores of men and total FSFI and subdomains of FSFI scores of women whom double-J stent was placed were evaluated before and after the procedure, there was statistically significant reduction in patient’s scores. In the control group, any statistically significant alteration in patient’s score was not observed in men and women. Sexual functions are negatively affected in both women and men whom ureteral stents are placed. To reduce these problems specific to urinary system owing to stents, new treatment strategies and new studies that lead to improvements in the material and design of stents are required.

The present study investigated the effects of stinging nettle (Urtica dioica L.) (UD) on benign prostatic hyperplasia (BPH) induced by testosterone. In vitro studies were conducted to assess the 5α-reductase inhibitory potential of UD. Two biochemical markers viz., β-sitosterol and scopoletin, were isolated and characterised in the extracts utilising High-performance thin layer chromatographic, FTIR, NMR and overlain UV spectral studies. Hyperplasia was induced in rats by subcutaneous administration of testosterone (3 mg kg⁻¹ s.c.) for 28 days in all the groups except the vehicle-treated group. Simultaneous administration of petroleum ether and ethanolic extracts (10, 20 and 50 mg kg⁻¹ p.o.) and isolated β-sitosterol (10 and 20 mg kg⁻¹ p.o.) was undertaken. Finasteride was used as a positive control (1 mg kg⁻¹ p.o.). Measurement of prostate/body weight ratio, weekly urine output and serum testosterone levels, prostate-specific antigen levels (on day 28) and histological examinations carried out on prostates from each group led us to conclude that UD can be used as an effective drug for the management of BPH.

The aims of this investigation were to test a novel technology comprising cryoprotectant-free vitrification of the spermatozoa of rainbow trout and to study the ability of sucrose and components of seminal plasma to protect these cells from cryo-injuries. Spermatozoa were isolated and vitrified using three different media: Group 1: standard buffer for fish spermatozoa, Cortland^® medium (CM, control); Group 2: CM + 1% BSA + 40% seminal plasma; and Group 3: CM + 1% BSA + 40% seminal plasma + 0.125 m sucrose. For cooling, 20-μl suspensions of cells from each group were dropped directly into liquid nitrogen. For warming, the spheres containing the cells were quickly submerged in CM + 1% BSA at 37 °C with gentle agitation. The quality of spermatozoa before and after vitrification was analysed by the evaluation of motility and cytoplasmic membrane integrity with SYBR-14/propidium iodide staining technique. Motility (86%, 81% and 82% for groups 1, 2 and 3, respectively) (P > 0.1) was not decreased significantly. At the same time, cytoplasmic membrane integrity of spermatozoa of Groups 1, 2 and 3 was changed significantly (30%, 87% and 76% respectively) (P < 0.05). All tested solutions can be used for vitrification of fish spermatozoa with good post-warming motility. However, cytoplasmic membrane integrity was maximal in Group 2 (CM + 1% BSA + 40% seminal plasma). In conclusion, this is the first report about successful cryoprotectant-free cryopreservation of fish spermatozoa by direct plunging into liquid nitrogen (vitrification). Vitrification of fish spermatozoa without permeable cryoprotectants is a prospective direction for investigations: these cells can be successfully vitrified with 1% BSA + 40% seminal plasma.

The role of leucocytospermia in male subfertility is a much debated topic despite being a frequent finding. This study aimed to identify the role of leucocytes, leucocyte subpopulations and natural killer cells in male subfertility. Seventy-sex subfertile men attending a regional andrology unit were recruited into this prospective study and subdivided into groups based on their semen analysis. The different leucocyte subpopulations were identified using immunocytochemical staining. Significant levels of CD3 helper T lymphocytes (P < 0.001) were present in the oligospermic, asthenospermic, oligoasthenospermic and obstructive azoospermic group compared to the normospermic group. Significant levels of B cells (P < 0.05) were present in the asthenospermic, oligoasthenospermic and obstructive azoospermic group. The natural killer cells (CD56) were significantly raised in the oligoasthenospermic and obstructive azoospermic group (P < 0.05). Our study suggests that leucocytospermia impairs sperm function through enhanced T helper cell modulation, increased B cell population which leads to increased levels of antisperm antibody and natural killer cells mediated sperm damage. The site of seminal leucocyte production is not necessarily confined to the vas or the epididymis.

Organophosphorus compounds are currently among the most frequently used pesticides worldwide, and therefore, the potential for human exposure to man is considerable. Their toxicity results in negative effects on many organs and systems such as the male reproductive system. So, vitamins that can offer spermatozoa protection are of great importance. This study was designed to investigate (i) the possibility of dimethoate, an organophosphate insecticide, to induce oxidative stress response in rat spermatozoa in vitro and its effect on antioxidant defence system and (ii) the role of vitamin C and vitamin E in alleviating the cytotoxic effects of dimethoate Epididymal spermatozoa were incubated for 3 h at 37 °C with different concentrations of dimethoate (50, 100 and 200 μm) without vitamins or pre-incubated with 20 mm of vitamin C or 2 mm of vitamin E. Sperm parameters, malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) levels were performed. Dimethoate caused a significant induction of oxidative damage in spermatozoa at different concentrations as evidenced by increased MDA levels. However, a significant decrease in sperm mobility, viability and activities SOD, CAT and GPx was observed. Vitamins pre-treated spermatozoa showed a significant protection against the cytotoxic effects induced by dimethoate on studied parameters.

Male fertility largely depends on sperm quality, which may be affected by environmental and genetic factors. Recent data emphasised the implication of the polymorphism of mitochondrial DNA polymerase gamma (POLG) CAG repeats in male infertility. In this report, we explored a possible role of the (POLG) gene polymorphism in male infertility in Tunisian men. The polymorphic CAG repeat in the nuclear POLG gene was studied in 339 male subjects (216 patients with infertility (69 azoospermic, 115 oligoasthenoteratospermic and 32 normospermic) and 123 fertile) after DNA amplification by PCR, followed by genotyping using an automatic sequencer. The heterozygous and the homozygous mutant genotypes (10/≠10 and ≠10/≠10) were significantly more frequent among infertile patients than among fertile controls (11.2% versus 1.6%, P = 1.3 × 10⁻³ and 4.6% versus 0.8%, P = 4.2 × 10⁻⁷ respectively). We also found a significant difference between the frequencies of 10/≠10 genotype in azoospermic (4.4%) and in oligoasthenoteratospermic (15.6%) infertile patients (P = 2.6 × 10⁻²). However, the homozygous mutant genotype (≠10/≠10) was seen at similar frequencies in azoospermic, normospermic and oligoasthenospermic men (4.4%, 3.1% and 5.2% respectively). Under our conditions, the findings showed an association between POLG CAG repeat polymorphism and male infertility in Tunisian population.

In this study, isolated spermatogonial stem cells (SSCs) and Sertoli cells using enzymatic digestion from patients with maturation arrest of spermatogenesis were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% foetal calf serum in three different groups: (1) SSCs cultured without Sertoli cells (2) SSCs co-cultured with Sertoli cells (as control group), (3) SSCs co-cultured with Sertoli cells and adding different concentrations of basic fibroblast growth factor (0.1, 1, 10 ng ml⁻¹) and human leukaemia inhibitory factor (1000, 1200, 1500 unit ml⁻¹) as experimental groups. The assessment of colonies every 10 days during 5-week cultures showed that in the first group, the average number and diameter of the colonies were significantly lower than in the other groups (P < 0.05). The largest number of colonies was observed in control condition (32.29 ± 9.15) in day 30. The largest diameter of colonies was formed in combination dosages of 1 ng ml⁻¹ basic fibroblast growth factor (bFGF) + 1500 unit ml⁻¹ leukaemia inhibitory factor (LIF) (302.93 ± 37.68) and 10 ng ml⁻¹ bFGF and 1200 unit ml⁻¹ LIF (262.87 ± 35.54) in day 30 respectively. Isolated SSCs were positive for spermatogonial cell markers such as Oct4, Stra8, Piwil2 and Vasa but negative for Nanog. Transplantation technique indicated that hSSCs have good efficiency for colonisation of mouse seminiferous tubules after proliferation in culture system.

The objective of the study was to assess whether seminal plasma total antioxidant status (TAS) can be used as a biochemical predictor of male fertility and its variation in different categories of infertile male subjects in our population. The study population consisted of 28 fertile and 127 infertile [teratozoospermic (30), asthenoteratozoospermic (30), azoospermic (21), oligoastheno-teratozoospermic (20), polyzoospermic (15) and oligozoospermic (11)] male subjects. Seminal plasma was separated by centrifugation and stored at minus 80 degree Celsius. Semen was analysed using computer-assisted semen analysis according to WHO criteria. Seminal plasma TAS was estimated by colorimetric method using Randox total antioxidant status kit. TAS of fertile male subjects was significantly (P < 0.001) higher than that of infertile patients. In whole studied population, seminal plasma TAS showed a significant positive correlation with sperm concentration (P < 0.001), sperm motility (P < 0.0001), and spermatozoa with normal morphology (P < 0.0001). In conclusion, this study suggests that TAS of seminal plasma is one of the important factors contributing to male infertility, and it can be used as a biochemical predictor for male fertility.

The aim of the present study was to evaluate the association of Glycine102Serine molecular variant of luteinising hormone (LH) β-subunit gene with infertility in Egyptian males and to assess the impact of oxidative stress in association with Glycine102Serine molecular variant on male infertility. Fifty men with idiopathic oligozoospermia were enrolled in the study. The control group consisted of 39 men with normal semen parameters and proven fertility. Variable oxidative status markers were evaluated. Glycine102Serine variant was investigated using restriction fragment length polymorphism-polymerase chain reaction. The study concluded that Glycine102Serine variant of LH β-subunit gene is relatively common among infertile Egyptian males and might be implicated in some Egyptian infertility cases. In addition, the association of this variant, in some cases, with disturbed oxidative status (decreased seminal plasma antioxidants’ level and increased sperm lipid peroxidation) probably have a significant role in accentuation of infertility in those particular cases.

We investigated the effects of quercetin on pathological findings on testicular ischaemia–reperfusion (I/R) injury in rats. Twenty-four male Wistar albino rats were randomly assigned into four groups: Group 1, control (n = 5); Group 2, sham (n = 4); Group 3, I/R (n = 8); and Group 4, I/R + quercetin (n = 7). Bilateral testicular artery and vein were occluded for 1 h, followed by reperfusion in I/R and I/R + quercetin animals. Quercetin (20 mg kg⁻¹ per day) was administrated once daily by gavage to Group 1 and Group 4, respectively, after reperfusion. At the end of the study, bilateral orchiectomies were performed for histopathologic examination. The tissue damage was evaluated with light microscopy. Normal inter-stitium and seminiferous tubules were observed in control group. In the sham group, rats were seen minimal oedema around the seminiferous tubules and congested vascular structures. In Group 3, oedema, vascular congestion and haemorrhage between seminiferous tubules were observed. In Group 4, histopathologic features were markedly less than Group 3 (P = 0.03). Our study demonstrated that quercetin seems to have a protective effect on testis histopathology in rats with testicular I/R.

A number of herbal drugs are advocated in the traditional Ayurvedic literature for the improvement of overall sexual function. Young roots of Bombax ceiba Linn. (Fam. Bombacaceae) [correction added after online publication 1 August 2011: the family of Bombax ceiba was incorrectly mentioned as Orchidaceae. It has been corrected to Bombacaceae] also known as Semal Musli are used traditionally in Indian subcontinent as sexual stimulant. Its juice is considered nutritive and restorative tonic. Lyophilised aqueous extract of roots was studied for effect on sexual behaviour and spermatogenesis in male albino rats. Administration of 100 mg Kg⁻¹ body weight of aqueous extract influenced the five parameters evaluated in vivo. Sexual behaviour analysis in the presence of a female rate, serum testosterone level, anabolic effects, epididymal sperm count and seminal fructose level were the parameters evaluated. In B. ceiba extract-treated animals, a gain in body and sexual organ weights was observed. Mount, intromission and ejaculation frequencies were significantly improved (P < 0.05). An increase in serum testosterone levels was also observed, but it was not statistically significant (P > 0.05). Seminal fructose content and epididymal sperm count were significantly improved as well. Penile erection index was also higher compared to control group animals. Hesitation time was significantly reduced (P < 0.01), and copulatory rate was doubled in treated animals compared with control group animals.

The aim of this study was to ascertain the variations in the reproductive sex ratio (number of men to number of women) among male professional basketball players in Spain. This retrospective, cross-sectional study is based on a survey conducted in the Spanish professional basketball leagues during the season 2009–2010. A total of 172 professional basketball players completed an anonymous survey. Forty-seven of the respondents had offspring, with a total of 61 children: 70% girls and 30% boys, with a sex ratio value of 0.42. Thirty-three basketball players were Caucasian (CAU), with 44 children, nine boys and 35 girls (sex ratio = 0.26). Fourteen were black, of African heritage (AFR), with 17 children, nine boys and eight girls, (sex ratio = 1.12). Differences (P < 0.01) were found in offspring sex ratio values for all basketball players (sex ratio = 0.42) and for CAU group (sex ratio = 0.26) when compared with the general Spanish population (sex ratio = 1.06). Moreover, a significant seasonal variation was observed in CAU offspring sex ratio during the first quarter compared with the rest of the year (0.66 versus 0.12) (P < 0.03). In conclusion, a significant increase in the sex ratio value in favour of female offspring was observed in the group of CAU professional basketball players.

The aim of this study was to administer to two groups of patients with male accessory gland infection (MAGI), respectively, with positive or negative alterations in ultrasonography, a new diagnostic interview, arbitrarily named structured interview about MAGI (SI-MAGI), to evaluate differences between these groups, especially about the prevalence of sexual dysfunction. After ultrasound examinations, patients with MAGI were divided into two age-matched groups: positive and negative for ultrasound signs (US+ and US−). The SI-MAGI was structured into four domains (urinary tract symptoms, ejaculatory pain or discomfort, sexual dysfunction and quality-of-life impact) for a total of 30 questions with four possible answers. Infertile patients of MAGI US+ group showed scores significantly higher than MAGI US− and healthy control group in all domains (anovaP < 0.005) in relation to scores of patients with MAGI US+ and US−: in domain 1 = 16.0 ± 0.5 vs 9.0 ± 0.5, domain 2 = 21.0 ± 1.0 vs 11.0 ± 1.0, domain 3 = 23.0 ± 0.5 vs 12.0 ± 1.0 and, finally, in domain 4 = 13.0 ± 2.0 vs 6.0 ± 1.0. In particular, a higher frequency of sexual dysfunction (52%) was detected in MAGI US+ group when compared with MAGI US− (28%). This study introduces a specific set of questions, which combined with the sperm analysis, microbiological and ultrasound investigations, that altogether better express the clinical presentation of MAGI. Finally, MAGI US+ group showed a high percentage of sexual dysfunction.

We evaluated the effects of vardenafil on testicular androgen-binding protein secretion (ABP). Bilaterally obstructed azoospermic (OA)-men (n = 19) (group A) underwent unilateral testicular biopsy. A group of nonobstructed azoospermic (NOA)-men (n = 68) (group B) underwent bilateral testicular biopsy. ABP secretion in vitro by testicular tissue was assessed in each participant of every group. In addition, intracytoplasmic sperm injection (ICSI) cycles were performed in several couples of group A or group B using frozen/thawed spermatozoa from the biopsy material. Ten OA-men (group A1), 14 NOA-men (group B1), and nine different NOA-men (group B2) had been positive for spermatozoa in the biopsy but pregnancies were not achieved in the respective female partners. Men of groups A1, B1 and B2 were treated with vardenafil, vardenafil and l-carnitine respectively. Then, the men of groups A1, B1 and B2 underwent a second testicular (unilateral) biopsy. Within the group A1 and within the group B1, ABP secretion rate was significantly larger after vardenafil treatment than prior to vardenafil treatment. In addition, fertilisation rates in ICSI cycles within groups A1 or B1 were not affected by vardenafil administration. Vardenafil administration in NOA-men increased ABP secretion and did not affect detrimentally the presence of testicular foci of advanced spermatogenesis.

The objective of research was to determine the sensitivity, specificity and positive predictive value of serum follicle stimulating hormone and testis size in predicting spermatogenesis in infertile men with azoospermia. In a prospective study, azoospermic men were studied. Serum follicle stimulating hormone measurement and scrotal sac ultrasonography were performed. Bilateral testis biopsy was performed for all of these patients. The sensitivity and specificity of follicle stimulating hormone and testis size were determined to predict the existence of different cellular steps of spermatogenesis. Of eighty infertile men who recruited into the study, 53 patients did not represent any different cellular steps of spermatogenesis, while 27 of them had various steps of such differentiation. Among the 53 patients without cellular steps of spermatogenesis in the biopsy, 41 were predicted to be azoospermic based on their serum follicle stimulating hormone levels (77.3% sensitivity), and of 27 patients with various cellular steps of spermatogenesis in the biopsy, 23 were predicted to have spermatozoa according to the follicle stimulating hormone level (85.2%) specificity. It is suggested that combination of these two indicators can substitute the invasive testis biopsy for predicting the existence of spermatozoa in infertile men with azoospermia.

Platelet activation results from the exposure of receptors on the surface of the platelet to specific structures on the vessel wall. The aim of this study was to assess the serum concentrations of apoptotic endothelial microparticles (EMPa) and the vitronectin receptor (VR) in patients with erectile dysfunction (ED) before and after treatment with tadalafil. This study included 50 patients with arterial ED. EMPa and VR levels were measured by using flow cytometry. The CD45_neg-CD144_pos-annexin V_Pos events were defined as EMPa, and the CD51_pos–CD61_pos events were defined as VR. Patients with ED were evaluated before and after daily treatment with 5 mg tadalafil for 90 days. Patients with arterial ED had serum concentrations of EMPa and VR significantly higher than the control group at baseline. After tadalafil, the serum concentrations of EMPa and VR of the patients with arterial ED were significantly lower than before treatment, but significantly higher than controls. Patients with arterial ED expressed higher levels of both EMPa that are associated with dysfunction of the arterial endothelial pathway and VR-expressing platelet-endothelial elements. Chronic treatment with tadalafil reduced endothelial apoptosis in these patients and also reduced VR expression.

This study aimed to compare the in vitro effects of coculture with Sertoli and SIM mouse embryo-derived thioguanine- and ouabain-resistant (STO) feeder layer cells on the efficiency of adult mouse spermatogonial stem cells (SSCs) colony formation. Sertoli and SSCs were isolated from testes, and their identity was confirmed using immunocytochemistry against Oct4, CDH1, PLZF and C-kit for SSCs and vimentin for Sertoli cells. SSCs were cultured in a simple culture system (control group) and on top of the Sertoli and STO feeder layers for 2 weeks. The number and diameter of colonies were evaluated during third, 7th, 10th and 14th day of culture, and the expression of the Oct-4, α6 and β1 integrins was assessed using quantitative RT-PCR. Significant differences were observed between the three groups, separately for each time (P < 0.05), with higher mean in number and diameter for Sertoli cells (P < 0.05). The results of RT-PCR showed higher gene expression of β1 integrin in Sertoli group, but no significant differences were observed in Oct-4 and α6 integrin gene expression among the three groups. Based the on the optimal effect of Seroli cells on the colony formation of SSCs, it is suggested to use these cells for better colonisation of SSCs.

Llama production in Argentina has increased, as the international interest in breeding this type of animals has grown in the last years. Considering the great polymorphism that llama spermatozoa present at evaluation using light microscopy, the aim of this study was to objectively evaluate llama sperm head morphometry using digital morphometric analysis. Five ejaculates from each of eight males were obtained to evaluate morphometric parameters of 8000 sperm heads stained with Tinción 15^®. The following average results were obtained for each parameter: size parameters: area 20.09 μm², length 6.60 μm, width 4.14 μm, equivalent circle diameter 5.06 μm, curve length 5.79 μm and curve width 3.48 μm; boundary parameters: perimeter 18.54 μm and convex perimeter 17.34 μm; and shape parameters: roundness 1.28 and elongation 1.59. Morphometric parameters of sperm head were compared between ejaculates of the same male and between males. Significant differences between ejaculates of the same male were found for all parameters evaluated (P < 0.01). Significant differences between males were found for all morphometric parameters (P < 0.01) except for curve length, curve width and perimeter. The differences detected would indicate that there is not a single morphometric pattern for Lama glama sperm head, because parameter values cannot be standardised.

A considerable proportion of male factor infertility cases are associated with inflammatory processes. The most common sexually transmissible agents causing sexually transmitted diseases in industrial countries are Chlamydia trachomatis, genital Ureaplasma and Mycoplasma spp. This study was undertaken to investigate whether these bacterial contaminants in semen affect sperm quality parameters and particularly DNA integrity (detected by sperm chromatin structure assay) in males from infertile couples (n = 293). The results showed that semen contaminations with the investigated bacterial species were not associated with sperm DNA fragmentation. However, contaminations with Mycoplasma spp. and C. trachomatis were associated with decreased sperm concentrations. Total sperm numbers in contaminated semen samples tended to be decreased, but not significantly. Mycoplasma had the highest adverse effect on sperm quality (concentration, motility, morphology and DNA condensation). Antibiotic therapy of the selected 47 men was successful in 55%, but semen quality parameters did not improve at least up to 3 months after the therapy. The presence of pathogenic bacteria in semen is primarily associated with low sperm production. Our data showed that Mycoplasma spp. contamination of semen had the highest adverse effect on sperm quality. Sperm chromatin integrity assessed by the presence of DNA breaks was not disturbed.

The buried penis syndrome in adults is a rare condition of different aetiologies. Today extreme obesity is considered as a major contributor. We present a case of a 30-year-old extremely obese patient (BMI 65 kg/m²) with purulent infection of the penile cavity, a phlegmon of the mons pubis and urinary retention due to a buried penis. Whereas acute complications of a buried penis in obese patients include local infection and urinary retention, chronic problems are undirected voiding, disturbed vaginal penetration and erectile dysfunction. Even though several surgical techniques are described, weight reduction should be primarily preferred.

It is well established that cystic fibrosis transmembrane conductance regulator gene (CFTR) mutations are involved in congenital bilateral absence of the vas deferens (CBAVD), causing obstructive azoospermia and male infertility. Also, several studies reported a relatively high prevalence of CFTR gene mutations in healthy men presenting reduced sperm quality. In this study, we investigate ΔF508 mutation and IVS8-polyT polymorphism in CFTR gene in Tunisian infertile men without CBAVD. Genetic analyses were performed in 148 infertile patients and 126 fertile individuals. The polymorphic IVS8-polyT tract in CFTR gene was analysed in only 129 infertile patients and 54 individuals of control group. As well, we screened for Y chromosome microdeletions in all infertile patients. No ΔF508 mutation was diagnosed either in infertile patients or in control group. 5T allele of IVS8-polyT tract was found in both infertile men (4.26%) and fertile individuals (8.33%). 5T/5T genotype was observed only in two azoospermic patients without Y microdeletions. The most frequent genotype of IVS8-polyT tract in infertile men and controls was 7T/7T (69.75% and 59.25% respectively). There was no association between IVS8-polyT polymorphism and reduced semen quality. Neither ΔF508 mutation nor 5T allele is involved in pathogenesis of male infertility in Tunisian infertile patients without CBAVD.

A 45-year-old male with azoospermia was presented to us with primary infertility. We found that he had been taking testosterone due to erectile dysfunction. Upon its discontinuation, the normozoospermia was restored. This led to a pregnancy. In men aged 40–50, the diagnostic work up of testosterone deficiency should be in accordance with the current guidelines. Moreover, no testosterone prescription should be made without having inquired about parenthood desires. In men of the reproductive age with recent-onset azoospermia, consider the possibility of an iatrogenic cause, primarily due to the testosterone replacement therapy. The treatment is simple: to discontinue the therapy.

The use of phosphodiesterase-5 inhibitors has become the first-line treatment of erectile dysfunction nowadays. The daily application of tadalafil has become a line of treatment of other diseases such as pulmonary hypertension and cardiomyopathy. This study aimed at exploring whether the chronic use of tadalafil has an adverse effect on the testes and semen of old albino rats. Sixty male albino rats were divided into three groups. Group 1: given 2 ml saline orally for 90 days. Group 2: received tadalafil orally (1.8 mg kg⁻¹ day⁻¹ for 3 months equivalent to 20 mg day⁻¹ for 3 months as in human dose) and Group 3: received tadalafil orally (1.8 mg kg⁻¹ day⁻¹ for 6 months equivalent to 20 mg day⁻¹ for 6 months as in human dose). Animals were observed daily for signs of toxicity and mortality. Body weight and food consumption were recorded once a week. After sacrificing the animals, gross examination of the testes was performed in situ and then the epididymis was processed for the evaluation of sperm parameters and testes with other organs relative weight was calculated. Testicular histopathological examination was performed to evaluate microscopic changes in the seminiferous tubules. The mean testicular weight was significantly lower in animals of Group 3, and no significant changes were observed in Group 2. Sperm count showed a significant time-dependent decrease. Sperm motility decreased significantly in both groups with higher effect in Group 3. Incidence of abnormal forms increased in both groups (about 5 and 7 times in Group 2 and 3 respectively). Histological examination revealed mild changes in Group 2 and moderate changes in Group 3 in the form of loosely packed connective stroma around seminiferous tubules, reduction in number of spermatogenic cells with sloughing of many spermatocytes within the lumen of some tubules. Large vacuoles appeared in the tubules which contained a fewer number of sperm. Sperm bundles were degenerated in most tubules and completely absent in others. It is concluded that chronic daily use of tadalafil produces detrimental effects on the structure and function of the testes of old male albino rats which are duration dependent.

Lepidium meyenii (maca) is a plant that grows exclusively above 4000 m in the Peruvian central Andes. Red maca (RM) extract significantly reduced prostate size in rats with benign prostatic hyperplasia (BPH) induced by testosterone enanthate (TE). Zinc is an important regulator of prostate function. This study aimed to determine the effect of RM on prostate zinc levels in rats with BPH induced by TE. Also, the study attempted to determine the best marker for the effect of RM on sex accessory glands. Rats treated with RM extract from day 1 to day 14 reversed the effect of TE administration on prostate weight and zinc levels. However, RM administered from day 7 to day 14 did not reduce the effect of TE on all studied variables. Finasteride (FN) reduced prostate, seminal vesicle and preputial gland weights in rats treated with TE. Although RM and FN reduced prostate zinc levels, the greatest effect was observed in TE-treated rats with RM from day 1 to day 14. In addition, prostate weight and zinc levels showed the higher diagnosis values than preputial and seminal vesicle weights. In conclusion, RM administered from day 1 to day 14 reduced prostate size and zinc levels in rats where prostatic hyperplasia was induced with TE. Also, this experimental model could be used as accurately assay to determine the effect of maca obtained under different conditions and/or the effect of different products based on maca.

In this experimental study, harmful effects of formaldehyde (FA) inhalation on sperm concentration, sperm quality, serum testosterone levels and the rat testes were investigated. In addition, the possible protective effects of rose oil against to these harmful effects were evaluated. For this purpose, 21 albino-Wistar rats were used. The rats in Group I were used as control group. When the rats of Group II were exposed FA (10 ppm/1 h) for 35 days, the rats of Group III inhalated rose oil (1 ml/1 h) after FA. The epididymal tissues were taken for sperm analysing and the testes were removed for histological examination. In addition, testosterone levels were determined from the blood samples. Although the testosterone levels, the epididymal sperm concentration, and the progressive sperm motility significantly decreased, the abnormal sperm rate significantly increased in the Group II when compared to Group I. In the Group III, these damages were seen less. When the rats in the Group II compared with the control group, there were serious histological damages. In the Group III, it was determined that the histological changes were less than group II. It can be expressed that serious damages occurred via formaldehyde exposure in male reproductive system and that the rose oil had protective effects against these damages.