Analysis of 29 Targeted Genes for Non-Obstructive Azoospermia: The Relationship between Genetic Testing and Testicular Histology.

PURPOSE: To analyze the presence of potentially pathogenic variants of 29 candidate genes known to cause spermatogenic failure (SPGF) in patients with non-obstructive azoospermia (NOA) who underwent testicular histology. MATERIALS AND METHODS: Forty-eight patients with unexplained NOA referred to the Department of Transfusion Medicine and Transplantation Biology, University Hospital Center Zagreb, Zagreb, Croatia for testicular biopsy. They were divided into three groups: those who had cryptorchidism (n=9), those with varicocele (n=14), and those with idiopathic NOA (n=25). All included patients underwent blood withdrawal for next-generation sequencing (NGS) analysis and gene sequencing. RESULTS: We found a possible genetic cause in 4 patients with idiopathic NOA (16%) and in 2 with cryptorchidism (22%). No pathogenic or possibly pathogenic mutations were identified in patients with varicocele. Variants of undetermined significance (VUS) were found in 11 patients with idiopathic NOA (44%), 3 with cryptorchidism (33%), and 8 patients with varicocele (57%). VUSs of the USP9Y gene were the most frequently as they were found in 14 out of 48 patients (29%). In particular, the VUS USP9Y c.7434+14del was found in 11 patients. They showed varied histological pictures, including Sertoli cell-only syndrome, mixed atrophy, and hypospermatogenesis, regardless of cryptorchidism or varicocele. No direct correlation was found between the gene mutation/variant and the testicular histological picture. CONCLUSIONS: Different mutations of the same gene cause various testicular histological pictures. These results suggest that it is not the gene itself but the type of mutation/variation that determines the testicular histology picture. Based on the data presented above, it remains challenging to design a genetic panel with prognostic value for the outcome of testicular sperm extraction in patients with NOA.

Effect of Green Tea on Weight Gain and Semen Quality of Rabbit Males.

The goal of the current study was to evaluate the action of the green tea plant (Camellia sinensis, L) on male rabbit reproduction and some non-reproductive indexes. Male rabbits were fed either a standard diet (control group) or a diet enriched with green tea powder (experimental groups; E): 5 g (E1) or 20 g (E2) per 100 kg of the milled complete feed mixture. Weight gain, sperm concentration, total and progressive motility, as well as haematological, and biochemical parameters and changes in testicular tissue histomorphology were evaluated. Feeding with green tea, at both tested concentrations, decreased weight gain per week and the total average weight gain compared to the control group (p < 0.05). Furthermore, green tea decreased sperm concentration, motility and progressive motility in the group fed with a lower dose (5 g) of green tea powder (p < 0.05), whilst a higher dose (20 g) was neutral. Some haematological and biochemical indexes, like medium-size cell count (MID), mean corpuscular haemoglobin concentration (MCHC), platelet percentage (PCT), levels of phosphorus (P) and total proteins (TP) were decreased in one or both experimental groups (p < 0.05), whilst the triglyceride level (TG) was increased in the E2 group (p < 0.05). The thicknesses of the testicular seminiferous tubules and epithelial layer were not affected by any concentration of green tea powder (p > 0.05). These observations suggest that green tea in the diet may have an adverse effect on rabbit growth and sperm quality, but their effect may be potentially dose-dependent.

Effects of quetiapine administration on sperm quality and testicular histology.

Quetiapine is one of the most commonly prescribed antipsychotics to treat schizophrenia in adults, in particular. In this study, quetiapine's effects were assessed on healthy sperm production in rats at repeated-pharmacological doses. Additionally, the effects of quetiapine on oxidative status and hormonal balance were also evaluated in rats. Quetiapine was administered to rats orally at 10, 20, and 40 mg/kg body weight doses for 28 days. At the end of this period, body and organ weights were measured, sperm concentration, motility, and morphology were determined, sperm damage was assessed, and histopathological analysis of testicular tissue was performed. Additionally, serum FSH, LH, and testosterone levels as male reproductive hormones were measured. Catalase, superoxide dismutase, glutathione, and malondialdehyde levels were determined for evaluating the oxidative status of testicular tissue. The findings obtained in this study showed that relative epididymis weights and sperm concentration decreased and abnormal sperm morphology increased in quetiapine-administered rats. Irregularity of typical architecture of the seminiferous tubules and germinal cell disorganization was observed in testicular sections of 20 and 40 mg/kg quetiapine-administered rats. Further, serum LH and testosterone levels decreased in 20 and 40 mg/kg quetiapine-administered rats. Additionally, decreased catalase and superoxide dismutase activities in testicular tissue of quetiapine-administered rats and increased malondialdehyde levels in testicular tissue of 40 mg/kg quetiapine-administered rats were measured. In conclusion, quetiapine treatment decreased sperm quality, altered hormone levels, and induced oxidative stress may be considered potential contributors to this adverse effect.

Vitamin B12 ameliorate Tramadol-induced oxidative stress, endocrine imbalance, apoptosis and NO/iNOS/NF-κB expression in Sprague Dawley rats through regulatory mechanism in the pituitary-gonadal axis.

This study aimed at the effect of vitamin B12 (VB12) on tramadol (TRM) induced pituitary-gonadal Axis toxicity. Thirty-two (32) adult male rats were randomized into four groups of eight (n = 8) rats each. Group A served as control was given 1 mL normal saline, group B received 50 mg /kg bwt TRM, group C received 0.5 mg/kg bwt VB12 and group D received 50 mg /kg bwt TRM and 0.5 mg/kg bwt VB12 through gastric gavage daily for 8 weeks. Parameters tested include sperm parameter, male reproductive hormone, testicular histology, glucose, lactate dehydrogenase (LDH), acid phosphate (ACP), and alkaline phosphate (ALP) activity, steroidogenic protein, cytochrome P450 A1, nitric oxide (NO), inducible nitric oxide synthase (iNOS), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), nuclear factor- kappa B (NF-κB), oxidative and antioxidant makers. Tramadol significantly decreases sperm quality, hormone, steroidogenic protein, cytochrome P450 A1, ACP, ALP, and increases glucose, LDH, oxidative stress, mtTFA, and UCP2, p53 expression, NO, iNOS, NF-κB, IL-1ß, IL-6, TNF-α, and caspase-3 activity. Degenerative alterations of the testes' and pituitary architecture and perturbation of spermatogenesis were observed in TRM-treated rats. The intervention of VB12 downregulated testicular oxidative stress, inflammatory markers, glucose, lactate, LDH, p53, caspase-3, mtTFA, and UCP2. And upregulate antioxidant, sperm quality, hormone, and spermatogenic cells. Vitamin B12 exhibited mitigation against TRM-induced testicular dysfunction via its antioxidant, anti-inflammatory and anti-apoptotic effects.

The impact of COVID-19 on the male reproductive tract and fertility: A systematic review.

OBJECTIVE: The COVID-19 pandemic, caused by the acute respiratory syndrome-coronavirus 2 (SARS-CoV-2), remains an ongoing public health challenge. Although males are affected slightly more than females, the impact of SARS-COV-2 on male reproductive system remains unclear. This systematic review aims to provide a concise update on the effects of COVID-19 on male reproductive health, including the presence of viral RNA in semen, and the impact on semen quality, testicular histology, testicular pain and male reproductive hormones. The global health is fronting an immediate as well as impending threat from the novel coronavirus (SARS-CoV-2) causing coronavirus disease (COVID-19), that inflicts more males than females. Evidence suggest that male reproductive system is susceptible to this viral infection. However, there are still several pertinent queries that remain to be fully explained regarding the mechanism in testicular SARS-CoV-2 dynamics and the exact mode of its actions. Thus, the present systematic review aims to provide a concise update on the effects of coronavirus disease 2019 (COVID-19) on male reproduction.. METHODS: A systematic review was conducted according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines searching the PubMed database. Eligibility for inclusion were original human studies evaluating the impact of COVID-19 on male reproductive health. Specific outcomes required for inclusion were at least one of the following: i) seminal detection of mRNA virus, or evaluation of ii) semen analysis, iii) testicular histology or ultrasonography, iv) testicular clinical symptoms and/or v) male reproductive hormones in COVID-19-positive patients. RESULTS: Of 553 retrieved articles, 25 met the inclusion criteria. This included studies primarily investigating the presence of viral RNA in semen (n = 12), semen quality (n = 2), testicular histology (n = 5), testicular pain (n = 2) and male reproductive hormones (n= 4). Results show little evidence for the presence of viral RNA in semen, although COVID-19 seems to affect seminal parameters, induce orchitis, and cause hypogonadism. Mortality cases suggest severe histological disruption of testicular architecture, probably due to a systemic and local reproductive tract inflammatory response and oxidative stress-induced damage. CONCLUSIONS: Clinical evaluation of the male reproductive tract, seminal parameters and reproductive hormones is recommended in patients with current or a history of COVID-19, particularly in males undergoing fertility treatment. Any long-term negative impact on male reproduction remains unexplored and an important future consideration.

Manipulation of fatty acid profiles in roosters' testes, alteration in sexual hormones, improvements in testicular histology characteristics and elevation sperm quality factor by L-carnitine.

The present study was designed to investigate the effects of different levels of l-carnitine (LC) on sperm quality factor (SQF), alterations in testis fatty acid profiles, testicular histology and reproductive hormones in young roosters. Eighteen broiler breeders (Ross 308) weighed at 3 months of age. They were randomly classified while each group had six birds. There were three experimental groups based on the LC concentrations (i.e. LC-0, LC-250, LC-500 mg per kg of diet). After two weeks of adaptation, semen samples were collected and evaluated for seminal attributes every two weeks (from week 24 to week 34). At the end of the experiments, four roosters from each treatment group were sacrificed in order to analyze testicular histology, testis fatty acid profiles and reproductive hormones. Supplementing the diet with two of the LC levels for 22 weeks caused significant rise in sperm concentration, viability and SQF compared to that of the control group (P < 0.05). Quadratic analysis in terms of number of seminiferous tubules and spermatogenesis index were significant (P<0.05). Tubular differentiation index improved linearly by the increasing levels of LC supplementation (P<0.01). The analysis of fatty acid profiles showed that LC significantly (P < 0.05) reduced the percentages of C14:0, C21:0, total saturated fatty acids, total odd-chain fatty acids and n-6/n-3 ratio. Moreover, LC significantly increased the percentage of C20:5n-3 (Eicosapentaenoic acid; EPA) (P < 0.05). Analysis of the correlation coefficient revealed that the SQF is in consistency with EPA (r = 0.98; P < 0.04). In contrast, SQF negatively and significantly correlates with odd-chain fatty acids (r = - 0.99; P < 0.001). The desaturation index for C16 fatty acids (16:1cis/C16:0) negligibly increased linearly as LC was added to the diet (P < 0.05). Furthermore, LC caused the roosters to have significant (P < 0.05) high levels of total testosterone and FSH concentrations. The concentration of LH in different treatment groups, however, turned out to be similar in response to the different levels of LC. In conclusion, long-term supplementation of rooster diet with LC can have beneficial effects on SQF and testis histology. The benefits include alterations in testicular histology, reproductive hormones and testicular fatty acid profiles.

Use of supplemental dietary coenzyme Q10 to improve testicular function and fertilization capacity in aged broiler breeder roosters.

In numerous studies it has been suggested that targeting mitochondria with specific compounds could efficiently inhibit various conditions associated with oxidative stress. The treatment of aged roosters with compounds such as coenzyme Q10 (CoQ10), may improve their reproductive performance by providing protection from oxidative stress. Therefore, this study was performed to assess the effect of supplemental dietary CoQ10 on the testicular function and fertility of aged broiler breeder roosters. A total of 36 roosters)47 weeks of age) were randomly divided into dietary treatments containing either 0, 300 or 600 mg CoQ10/kg diet. Three birds were allocated to each of four replicate groups in each dietary treatment. Between 47 and 54 weeks of age, ejaculates were obtained weekly from the three roosters in each replicate group. Samples in a replicate were pooled and analyzed as a single sample. Between 51 and 54 weeks of age, seminal plasma total antioxidant capacity (TAC), alanine amino transferase (ALAT) and aspartate amino transferase (ASAT) levels were assessed. Fertility, hatchability, and sperm penetration (SP) rates were likewise evaluated. Seminal volume, sperm concentration, sperm plasma membrane functionality, sperm plasma membrane integrity, seminiferous tubule diameter and seminiferous epithelium thickness exhibited quadratic increases in response to increasing levels of dietary CoQ10. Respectively, the 429.19, 433.33, 464.50, 613.50, 392.78 and 447.99 mg/kg dietary concentrations of CoQ10 provided the best results for each of the aforementioned variables. Also, other seminal traits, as well as testosterone concentration, fertility, and SP rates, displayed linear increases in response to the increasing levels of CoQ10. Dietary supplementation of CoQ10 linearly decreased seminal plasma ALAT and ASAT and linearly increased seminal plasma TAC. In conclusion, CoQ10 supplementation in the diet (a minimum of 300 mg CoQ10/kg diet) has the potential to improve the reproductive performance of aged broiler breeder roosters.

Reproductive toxic effects and possible mechanisms of zonisamide in male rats.

Zonisamide (ZNS) is an anticonvulsant which is used to treat the symptoms of epilepsy. Although it is frequently used during reproductive ages, studies that investigated the effects of ZNS on reproductive system are limited. Therefore, we aimed to assess the effects of ZNS on male reproductive system by oral administration to rats in 25, 50, and 100 mg/kg doses for 28 days. After the exposure period, sperm concentration, motility, morphology, and DNA damage, as biomarkers of reproductive toxic effects, were determined, and histopathological examination of testis was performed. In addition, levels of the hormones that play a role in the regulation of reproductive functions, such as follicle-stimulating hormone, luteinizing hormone (LH), and testosterone were measured and the levels of oxidative stress biomarkers that take part in the reproductive pathologies such as catalase, superoxide dismutase, glutathione, and malondialdehyde, were determined. Reproductive toxic effects related to ZNS administration were shown by the significant decrease of sperm concentration and normal sperm morphology in ZNS groups. Additionally, pathological findings were observed in the testicular tissues of ZNS-administered groups dose dependently. In addition, serum LH and testosterone levels were significantly decreased in the ZNS groups. Decreased catalase activities and increased malondialdehyde levels in ZNS groups were evaluated as oxidative stress findings in the testis tissue. It could be expressed that ZNS administration induced dose-dependent reproductive toxic effects in rats, and pathological findings associated with the reproductive system could be the result of that hormonal changes and testicular oxidative stress, which in turn might be considered as possible mechanisms of male reproductive toxicity.

Gonadal function and testicular histology in males with Prader-Willi syndrome.

CONTEXT: Cryptorchidism is common in Prader-Willi syndrome (PWS) males, but the testicular histology in childhood remains uncertain. The association between testicular histology and long-term gonadal function in PWS males is also unknown. OBJECTIVES: To evaluate the relationship between testicular histology in childhood and long-term gonadal function in PWS males. PATIENTS AND METHODS: Forty men with PWS were assessed longitudinally at our institute over the past 24 years. Clinical examinations and blood tests for LH, FSH and testosterone levels were compared with normal reference values. Tissue specimens were collected during orchiopexy and analyzed based on Nistal categories. RESULTS: Of nine testes available for pathological assessments, two showed favourable histology (Nistal I) and seven showed unfavourable histology (Nistal II or III). Of five postpubertal males with histology available, four reached puberty spontaneously, but only one reached Tanner stage 5. In a male with favourable histology, LH and FSH were high, but testosterone was normal, though below the average of the reference range. In three males with unfavourable histology, LH was normal, but FSH was highly elevated, and testosterone was at the lower limit of normal. One patient took hCG treatment to induce puberty; this patient showed favourable histology, but LH, FSH and testosterone were not elevated in adolescence. CONCLUSIONS: Testicular histology of PWS men in childhood varies from normal to Sertoli Cell-Only Syndrome. Regardless of the testicular histology in childhood, hypogonadism in PWS adults arises as a consequence of primary testicular dysfunction with highly elevated FSH and insufficient testosterone levels.

Relevance of testicular histopathology on prediction of sperm retrieval rates in case of non-obstructive and obstructive azoospermia.

INTRODUCTION: The aim of our research was to establish the relevance of testicular histopathology on sperm retrieval after testicular sperm extraction in patients with non-obstructive azoospermia and in patients with obstructive azoospermia, who already underwent a previous failure testicular fine needle aspiration. METHODS: We evaluated a total of 82 azoospermic men, underwent testicular sperm extraction, referring to the Assisted Reproductive Technology Centre of the University of Florence, Italy between January 2008 and March 2017. A general and genital physical examination, scrotal and trans-rectal ultrasound, semen analysis, hormone measurements, including follicle-stimulating hormone, luteinizing hormone and total testosterone, were collected. RESULTS: Successful sperm retrieval was obtained in 36 men of total (43.9%). Successful sperm retrieval was 29.5% in non-obstructive azoospermia patients, while men with obstructive azoospermia, who, underwent a previous failure testicular fine needle aspiration, had sperm retrieval in 86% of cases. Mean luteinizing hormone was 6.55 IU/L, total testosterone 4.70 ng/mL, right testicular volume 13.7 mL and left testicular volume 13.6 mL. Mean Follicle-stimulating hormone was 13.45 IU/L in patients with negative sperm retrieval and 8.18 IU/L in men with successful sperm retrieval. According to histology, 20.7% had normal spermatogenesis, 35.3% hypospermatogenesis, 35.3% maturation arrest and 8.5% Sertoli cell-only syndrome. Successful sperm retrieval was 88.2% in patients with normal spermatogenesis, 24.1% in the maturation arrest group and 48.27% in patients with hypospermatogenesis, while negative sperm retrieval was reported in Sertoli cell-only syndrome patients. Seven cases with maturation arrest showed a successful sperm retrieval. CONCLUSION: Testicular histopathology after testicular sperm extraction offers important information on prediction of sperm retrieval and can guide the surgeon in choosing the more suitable therapeutic practice.

Diabetes mellitus- induction: Effect of different streptozotocin doses on male reproductive parameters.

Diabetes mellitus (DM) is reported to be involved in male reproductive impairment, and its impact is evident in the increased prevalence of infertility. Various studies have reported that a single parenteral injection of <40 mg/kg Streptozotocin (STZ) is ineffective in ablating pancreatic ß-cells and creating a rat model to investigate the effect of DM on the male reproductive system. This study therefore aims to validate these claims. Adult male Wistar rats received either a single intraperitoneal injection of STZ (30 mg/kg or 60 mg/kg) or saline (0.9%, Control). Diabetes was confirmed after 72 h if plasma glucose levels were ≥14 mmol/L. Body weight, glucose level, fluid and food intake were measured weekly. Animals were sacrificed after 8 weeks of treatment by an overdose of sodium pentobarbital (160 mg/kg body weight). The testis and epididymis were harvested and weighed prior to preparation for histological evaluation. Epididymal sperm morphology was analysed using computer aided sperm analysis (CASA). STZ60 animals presented with significantly lower body weights compared to both control and STZ30 groups. Animals in both STZ30 and STZ60 groups showed decreased normal sperm morphology compared to control. Histological evaluation of the testes showed a decrease in the number of spermatozoa in the seminiferous tubules of animals in the STZ30 and STZ60 groups compared to control. A complete absence of spermiogenesis was observed in the seminiferous tubules of STZ60 animals. These findings prove that an STZ concentration of 30 mg/kg, which is much lower than the reported 40 mg/kg, has adverse effects on the male reproductive system via its diabetogenic effect and can therefore be used to study the impact of DM on male fertility.

Sulfur dioxide inhalation lowers sperm quality and alters testicular histology via increasing expression of CREM and ACT proteins in rat testes.

Sulfur dioxide (SO2) is one of the main atmospheric pollutants worldwide, and is reported to be responsible for the formation of severe haze in China. Some studies have demonstrated a potential harmful effect of SO2 on the male reproductive system; however the underlying mechanism is still unknown. The purpose of this study is to investigate the roles of cytochrome P450 (P450), cAMP-responsive element modulator (CREM), and activator of CREM (ACT) in SO2-induced toxicity. Forty-eight male Wistar rats were randomly divided into an experimental and control group. The experiment group was exposed to SO2 in ambient air (10ppm, 4h/day), and the control group was treated with filtered air in the same conditions. After 2 weeks, the results showed a significant decrease in body weight and sperm motility, and an increase in the testis weight-to-body weight ratio as compared to the control group. Histological investigation suggested that SO2 exposure led to loose arrangement of the spermatogenic cells and local structural damage in the seminiferous tubules. Moreover, the expressions of P450, CREM and ACT proteins increased in the testes by 0.22%, 47.26% and 23.38%, respectively. Taken together, SO2 inhalation lowered sperm quality, altered testicular histology, and increased expressions of CREM and ACT proteins in the testes of rats. Overall, these results could contribute to a better understanding of SO2-induced male reproductive toxicity.

Impact of testicular histopathology as a predictor of sperm retrieval and pregnancy outcome in patients with nonobstructive azoospermia: correlation with clinical and hormonal factors.

In this study, our objective was to evaluate the impact of testicular histopathology on the outcome of intracytoplasmic sperm injection (ICSI) cycles of patients with nonobstructive azoospermia and correlate with clinical and hormonal parameters. For this purpose, 271 patients with nonobstructive azospermia (NOA) who underwent testicular sperm extraction (TESE) for ICSI cycles were retrospectively evaluated for sperm retrieval, fertilisation, embryo cleavage, clinical pregnancy and live birth rates among different testicular histology groups. We also correlated hormonal and clinical factors with histological findings. Sperm retrieval and fertilisation rates (FR) were found to be significantly different among all testicular histological groups of NOA except for embryo cleavage, clinical pregnancy and live birth rates. Furthermore, serum follicle stimulating hormone (FSH) level was the most significant variable to predict sperm recovery on TESE. Separate analyses within each testicular histological group revealed that higher FSH was also associated with lower pregnancy rates in only maturation arrest group. In conclusion, testicular histology significantly influences sperm retrieval and FRs but not pregnancy and live birth rates in nonobstructive azoospermia. However, FSH is the best predictor of a successful TESE.

Evaluations of cytotoxicity of Smilax myosotiflora and its effects on sexual hormone levels and testicular histology in male rats

Objective: To investigate the cytotoxicity of Smilax myosotiflora (S. myosotiflora) methanolic extract and its effects on sexual hormone levels and testicular histology in male rats. Methods: The cytotoxicity of S. myosotiflora methanolic extract was investigated by employing brine shrimp lethality assay. Forty eight male rats were randomly divided into four groups (Groups I-IV) of 12 each. Rats in Group I were administered with 0.5 mL of distilled water (vehicle), whilst Groups II, III and IV received 200, 400 and 800 mg/kg of the methanolic extract of S. myosotiflora in 0.5 mL of the vehicle, respectively. Male rats treated with continuous daily dosing were killed and necropsied after a total dose period of 60 days. Sexual hormones were assayed and histological examination of testes was performed according to standard methods. Results: S. myosotiflora extracts did not produce any cytotoxicity to brine shrimp in all concentrations tested. Serum testosterone level was significantly higher in rats treated with high dose of S. myosotiflora. Testicular histology showed normal architecture with all stages of spermatogenesis in all experimental groups. Conclusions: The present work confirmed that S. myosotiflora extract improves reproductive functions, without any cytotoxic activity and produces no histological changes to the testes.

MORFOHISTOLOGÍA TESTICULAR DE Hypsiboas pulchellus (AMPHIBIA, HYLIDAE)DURANTE LA ESTACIÓN REPRODUCTIVA

En este estudio se aportan datos sobre la morfohistología testicular de Hypsiboas pulchellus durante el período reproductivo con el fin de conocer aspectos de su biología reproductiva. Los testículos, ovoides y amarillentos de 4,64 ± 0,48 mm de largo por 2,05 ± 0,23 mm de ancho, están rodeados por la túnica albugínea que mide 5,60 ± 0,82 µm. Internamente presentan lóculos seminíferos de 257,47 ± 58,25 µm de diámetro, distinguiéndose en ellos cistos con células espermatogénicas asociadas a células de Sertoli. Existe escaso tejido intersticial. Las espermatogonias I son las células más grandes de la serie germinal (14,34 ± 1,74 µm), estas originan a las espermatogonias II (10,14 ± 1,33 µm). Los espermatocitos I presentan la cromatina levemente condensada midiendo 9,34 ± 0,32 µm. Los espermatocitos II miden 8,12 ± 1,07 µm. Las espermátidas I son esféricas, miden 7,61 ± 1,45 µm, y se agrupan en cistos redondeados. Las espermátidas II, en cambio, son alargadas, no hallándoselas dentro de cistos (4,09 ± 0,51 µm). Los espermatozoides, alargados y flagelados, se hallan libres en el centro del lóculo. La morfohistología de los testículos analizados muestra características similares a las observadas en otros anfibios neotropicales, y presentan todas las células del linaje espermatogénico en un mismo lóculo.

In this study, data concerning to the morphological testicular features of Hypsiboas pulchellus during the breeding season is provided, in order to know aspects of their reproductive biology. The testis, ovoid and yellowish with 4.64 ± 0.48 mm length and 2.05 ± 0.23 mm wide, are surrounded by the tunica albuginea which measures 5.60 ± 0.82 µm. The seminiferous locules, placed internally, measure 257.47 ± 58.25 µm in diameter, and cysts with spermatogenic cells associated with Sertoli cells are distinguished in them. The interstitial tissue is scarce. The biggest cells of the germinal series are the spermatogonia I (14.34 ± 1.74 µm), from which spermatogonia II (10.14 ± 1.33 µm) originate. Spermatocytes I measure 9.34 ± 0.32 µm and have slightly condensed chromatin. The spermatocytes II are 8.12 ± 1.07 µm long. The spermatids I are spherical, grouped in rounded cysts, and measure 7.61 ± 1.45 µm. On the other hand, spermatids II are elongated (4.09 ± 0.51 µm) and not within the cysts. Towards the center of the loculus, the free, lengthened and flagellated spermatozoids are located. The morphohistology of the analyzed testicles show similarities with those observed in other neotropical amphibians, being all the cells of the spermatogenic lineage in the same locule.

Morphometric evaluation of seminiferous tubule and proportionate numerical analysis of Sertoli and spermatogenic cells indicate differences between crossbred and purebred bulls.

AIM: The present study compared the testicular cytology and histology between crossbred (Holstein-Friesian [HF] × Tharparkar) and purebred (HF and Tharparkar) bulls to find out differences if any. MATERIALS AND METHODS: Four peripubertal bulls from each breed were utilized for the study. Through percutaneous needle aspiration biopsy, Sertoli and spermatogenic cells were extracted, and morphometry was studied. For histological studies, testicular tissues obtained through unilateral castration were utilized. Sertoli cells specific GATA4 antibody was used to study the population of Sertoli cells in the seminiferous tubule through immunofluorescence. RESULTS: The testicular weight, volume, and scrotal circumference differed significantly among the breeds. The diameter and area of the seminiferous tubule was high in HF, followed by Karan Fries (KF), and Tharparkar bulls. However, the degree of compactness, based on qualitative evaluation, was high in Tharparkar followed by KF and HF bulls. The intensity of Leydig cells was higher in Tharparkar bulls followed by KF and HF. The proportion of Sertoli cells was higher (p<0.05) in HF and Tharparkar bulls compared to KF bulls. CONCLUSION: It may be concluded that variations exist in testicular components of the breeds studied and the proportion of Sertoli cells in relation to spermatogenic cells was significantly lower in crossbred bulls compared to purebred bulls.

Effects of chronic tramadol administration on testicular tissue in rats: an experimental study.

UNLABELLED: In a prospective experimental study, the effects of chronic tramadol administration on gonadotrophic and sex hormones, histopathological and morphometrical alterations in rat testicular tissue were investigated in a laboratory setting. Tramadol was given alone to adult male albino rats. Gonadotrophic and serum sex hormone levels were measured and testicular pathological and morphometric changes were observed in treated vs. CONTROL GROUP: After 30 days of treatment, tramadol induced a decrease in LH, FSH and testosterone serum levels. Histologically, degenerative changes in the seminiferous tubules were observed. They showed shrinkage, separation of tubular basement membrane, disorganisation and vacuolisation of spermatogenic layers. Morphometric analysis revealed significant decrease in the mean values of the tubular diameter and epithelial height. Ultrastructural abnormalities were detected in all cells of spermatogenic lineage in addition to the appearance of apoptotic cells. Sertoli cell vacuolation, huge lipid droplets and disrupted Sertoli cell junctions were observed. Leydig cells showed euchromatic nuclei and dilated smooth endoplasmic reticulum. In view of these findings, it is concluded that tramadol induces alterations in sex hormonal levels in conjunction with disruption of the normal histological structure of rat testis. This might lead to the risk of male infertility. Therefore, tramadol should be used with caution with appropriate dose monitoring.

Effects of the commercial antiandrogen flutamide on the biomarkers of reproduction in male Murray rainbowfish (Melanotaenia fluviatilis).

The endocrine responses in male Murray rainbowfish (Melanotaenia fluviatilis) were evaluated after exposures to biologically active concentrations of the nonsteroidal pharmaceutical, flutamide. Fish were exposed to nominal concentrations of 125 µg/L, 250 µg/L, 500 µg/L, and 1000 µg/L of flutamide for 7 d, after which plasma vitellogenin concentration; brain aromatase activity; and hepatic expression of the genes for vitellogenin, choriogenin, and androgen and estrogen receptors were assessed. Qualitative assessment of the testes of the fish exposed to flutamide exhibited hindrance in the transformation of spermatogonia to spermatozoa and increased testicular anomalies, such as multinucleated and pyknotic cells and interstitial fibrosis. An increase in the hepatosomatic index with respect to the controls was noted after treating the fish with flutamide at all concentrations. Vitellogenin was induced in plasma in the 1000 µg/L flutamide group. The activity of aromatase in the brain declined significantly after exposures to flutamide at all concentrations. Males exposed to 1000 µg/L of flutamide showed a downregulation in the genes encoding androgen receptors α and ß. The expression of the gene for the estrogen receptor α was induced and of vitellogenin was downregulated after treatment with 250 µg/L to 1000 µg/L of flutamide. The results suggest that 7-d exposures to 125 µg/L to 1000 µg/L flutamide can impair the reproductive endocrine system in male Murray rainbowfish at multiple levels by an antiandrogenic mode of action.

Azoospermia y fertilización asistida: rol de la histología testicular en el éxito de icsi / Azoospermia and assisted reproduction: role of testicular histology in the success of ICSI

Introducción: La infertilidad masculina afecta aproximadamente al 7por ciento de los hombres, presentándose hasta el 15 por ciento de ellos con azoospermia. El conocimiento del tipo de azoospermia (obstructiva o no obstructiva) y la localización de la falla (pre-testicular, testicular o post-testicular) es vital para conocer el pronóstico de fertilidad de la pareja y plantear un plan terapéutico adecuado. Actualmente, la extracción de espermatozoides desde epidídimo o testículo de pacientes azoospérmicos, y la posterior inyección intracitoplásmática de éstos (ICSI, por sus siglas en inglés) ha permitido obtener embriones viables para su posterior transferencia. Materiales y métodos: Estudio descriptivo retrospectivo de 42 parejas infértiles con diagnóstico de azoospermia; que se sometieron a biopsia testicular, ICSI y posterior transferencia de embriones, entre los años 2004 y 2012. Se lleva a cabo un análisis de la edad de los pacientes, resultados de la histopatología testicular y su asociación con los resultados de la fertilización asistida. Resultados: 42 pacientes azoospérmicos se sometieron a biopsia testicular y extracción de espermatozoides en el mismo acto quirúrgico. La edad promedio de los pacientes fue de 36 años para los hombres y 32 años para las mujeres. En el análisis histológico de los tejidos testiculares, el 31por ciento de los pacientes presentaban espermatogénesis conservada (EC), el 35.7 por ciento atrofia mixta (AM), el 14.3 por ciento hipoespermatogénesis (HE), el 14.3 por ciento detención de la maduración (DM) y un 4.8 por ciento presentaba otras histologías. Lograron embarazo clínico 14 de 42 parejas (33,3 por ciento), siendo la tasa de embarazo específica por patología de 38,5 por ciento para EC, 26.7 por ciento para AM, 50 por ciento para HE, 16,7 por ciento para DM y 50 por ciento para las otras histologías. 12 de las 42 parejas realizaron más de un ciclo de transferencias...

Introduction: Male infertility affects approximately 7percent of men, presenting up to 15 percent with azoospermia. Knowing the type of azoospermia (obstructive or non-obstructive) and the location of the problem (pre-testicular, testicular and post-testicular) is vital to recognize the fertility prognosis of the couple and present a proper treatment plan. Currently, the extraction of sperm from epididymis or testis of azoospermic patients, and subsequent intracytoplasmic sperm injection (ICSI) has yielded viable embryos for transfer. Materials and Methods: Retrospective study of 42 infertile couples diagnosed with azoospermia, who underwent testicular biopsy, ICSI and subsequent embryo transfer, between 2004 and 2012. We performed an analysis of the patients’ age, testicular histopathology results and their association with assisted fertilization outcome. Results: 42 azoospermic patients underwent testicular biopsy and sperm extraction in the same surgery. The average age of patients was 36 years for men and 32 years for women. Histologic analysis of testicular tissue showed that 31 percent of patients had normal spermatogenesis (NS), 35.7 percent mixed atrophy (MA), 14.3 percent hypospermatogenesis (HS), 14.3 percent maturation arrest (MTA) and 4.8 percent had other histologies. 14 of 42 couples achieved clinical pregnancy (33.3 percent), with a specific pregnancy rate of 38.5 percent for NS, 26.7 percent for MA, 50 percent for HS, 16.7 percent for MTA and 50 percent for other histologies. 12 of 42 couples underwent more than one transfer cycle. Conclusions: The use of ICSI is a suitable alternative for infertile couples with severe male factor. The results of this technique are favorable for most histologies. Patients with MA exhibit sertoli solo syndrome and / or tubular sclerosis foci associated to regions with normal spermatogenesis, enabling the sperm extraction for ICSI.

Histological and hormonal testicular function in oligo/azoospermic infertile men.

We characterised and correlated the histological and hormonal aspects of a cohort of 261 azo/oligozoospermic men, applying a quantitative/qualitative evaluation of testicular tissue and serum and intratesticular hormonal measurements. One hundred and 93 azo/oligozoospermic patients were diagnosed as: complete sertoli cell only syndrome (cSCOS), n = 76; focal SCOS, n = 31; maturation arrest, n = 34; hypospermatogenesis, n = 17; mixed atrophy, n = 25; and severe atrophy, n = 10. Normal spermatogenesis was observed in 68 infertile men (controls). Patients with cSCOS, focal SCOS, mixed and severe atrophy had larger LC/clusters (11.5; 11.0; 10.7; 18.9 LC/cluster) than controls (6 LC/cluster; P < 0.001). cSCOS, focal SCOS, mixed and severe atrophy patients had higher FSH, LH and lower T/LH ratio serum levels than the other groups. Intratesticular testosterone concentrations were higher in tissues with complete or focal SCOS (45.6 ng mg(-1) protein) and mixed atrophy (79.0 ng mg(-1) protein) than normal tissues (20.3 ng mg(-1) protein; P = 0.03 and P = 0.007). Considering all subjects, significant correlations were found between T/LH ratio and Leydig cells/cluster (r = 0.510, P < 0.001), FSH levels (r = -0.692, P < 0.001) and with intratesticular testosterone (r = -0.354, P = 0.001); these correlations follow the pattern of severity of spermatogenic damage. By a thorough histological evaluation, we validate the concept that the severity of spermatogenic impairment is associated with major morphological and functional disturbance of the Leydig cell compartment.