Absence Of Sperm Research Articles

Biallelic variants in MAATS1 encoding CFAP91, a calmodulin-associated and spoke-associated complex protein, cause severe astheno-teratozoospermia and male infertility

BackgroundMultiple morphological abnormalities of the flagella (MMAF) consistently lead to male infertility due to a reduced or absent sperm motility defined as asthenozoospermia. Despite numerous genes recently described to be...

Evaluating genetic causes of azoospermia: What can we learn from a complex cellular structure and single-cell transcriptomics of the human testis?

Azoospermia is a condition defined as the absence of spermatozoa in the ejaculate, but the testicular phenotype of men with azoospermia may be very variable, ranging from full spermatogenesis, through arrested maturation of germ cells at different stages, to completely degenerated tissue with ghost tubules. Hence, information regarding the cell-type-specific expression patterns is needed to prioritise potential pathogenic variants that contribute to the pathogenesis of azoospermia. Thanks to technological advances within next-generation sequencing, it is now possible to obtain detailed cell-type-specific expression patterns in the testis by single-cell RNA sequencing. However, to interpret single-cell RNA sequencing data properly, substantial knowledge of the highly sophisticated data processing and visualisation methods is needed. Here we review the complex cellular structure of the human testis in different types of azoospermia and outline how known genetic alterations affect the pathology of the testis. We combined the currently available single-cell RNA sequencing datasets originating from the human testis into one dataset covering 62,751 testicular cells, each with a median of 2637 transcripts quantified. We show what effects the most common data-processing steps have, and how different visualisation methods can be used. Furthermore, we calculated expression patterns in pseudotime, and show how splicing rates can be used to determine the velocity of differentiation during spermatogenesis. With the combined dataset we show expression patterns and network analysis of genes known to be involved in the pathogenesis of azoospermia. Finally, we provide the combined dataset as an interactive online resource where expression of genes and different visualisation methods can be explored ( https://testis.cells.ucsc.edu/ ).

The genetic architecture of morphological abnormalities of the sperm tail.

Spermatozoa contain highly specialized structural features reflecting unique functions required for fertilization. Among them, the flagellum is a sperm-specific organelle required to generate the motility, which is essential to reach the egg. The flagellum integrity is, therefore, critical for normal sperm function and flagellum defects consistently lead to male infertility due to reduced or absent sperm motility defined as asthenozoospermia. Multiple morphological abnormalities of the flagella (MMAF), also called short tails, is among the most severe forms of sperm flagellum defects responsible for male infertility and is characterized by the presence in the ejaculate of spermatozoa being short, coiled, absent and of irregular caliber. Recent studies have demonstrated that MMAF is genetically heterogeneous which is consistent with the large number of proteins (over one thousand) localized in the human sperm flagella. In the past 5 years, genomic investigation of the MMAF phenotype allowed the identification of 18 genes whose mutations induce MMAF and infertility. Here we will review information about those genes including their expression pattern, the features of the encoded proteins together with their localization within the different flagellar protein complexes (axonemal or peri-axonemal) and their potential functions. We will categorize the identified MMAF genes following the protein complexes, functions or biological processes they may be associated with, based on the current knowledge in the field.

The potential cytoprotective effect of Vitamin C and Vitamin E on monosodium glutamate-induced testicular toxicity in rats

ABSTRACT Background Monosodium glutamate (MSG) has been recognized as flavor enhancer that adversely affects male reproductive systems. Objective The study was conducted to explore the conceivable protective effects of vitamin C and/or vitamin E on testicular toxicity induced by MSG in rats. Materials and Methods Thirty male Wistar albino rats were divided (six per group) into: control, MSG, MSG + Vitamin C, MSG + Vitamin E and MSG + Vitamin C + Vitamin E groups. The duration of the study was three weeks. Assessment of serum testosterone, leuteinizing hormone (LH), malondialdehyde (MDA), glutathione peroxidase (GPX), interleukin-10 (IL-10) ,and tumor necrosis factor (TNF-α) were done. Histopathological examination of the testes of the rats was performed using histological, histochemical (Periodic Acid Schiff reaction (PAS)), and immunohistochemical (Proliferating cell nuclear antigen (PCNA), androgen receptors (ARs), Caspase-3) techniques. Results MSG-group was associated with significant decrease in serum testosterone, LH, GPX, and IL-10 (P < 0.05) and significant increase in serum MDA and TNF-α (P < 0.05) when compared with control group. MSG-group revealed many histopathological changes in the testis including degeneration of the germinal epithelium, absence of sperms in the lumina of tubules, widened vacuolated interstitium, marked deposition of the collagen fibers, very strong PAS reaction and marked immunohistochemical changes. Administration of vitamin C or vitamin E significantly reduced these changes; however, the combination of vitamin C and vitamin E provided more potent protection against the toxic effect of MSG than using each vitamin alone. Also, there was insignificant difference (P > 0.05) between MSG +Vitamin C and MSG +Vitamin E groups. Conclusion Vitamin C and Vitamin E act synergistically in reducing MSG-induced testicular toxicity via antioxidant, anti-inflammatory and antiapoptotic effects of both vitamins.

Experiences of Women with Infertility attending Gynae OPD of Tertiary Hospital of Rupandehi

INTRODUCTION: One of the most important and underappreciated reproductive health problems in developing countries is the high rate of infertility and childlessness. The inability to have a child is frequently considered a personal tragedy affecting the entire family and even the local community.&#x0D; MATERIAL AND METHODS: The descriptive phenomenological research design was used to explore experiences of women with infertility. Fifteen women with infertility were selected by purposive sampling technique. Data was collected through in-depth interview method using unstructured questionnaire and thematic analysis was done.&#x0D; RESULTS: Among 15, 46.66% of the participants had gynecological problems and 53.33% had medical problems such as hypothyroidism and hyperthyroidism. Thirty-three percentage of participants' spouse had problem of low sperm count and absence of sperm. Most of the participants and their spouses were under treatment. Sixty percentage of the participants had experienced psychosocial problems and 26.66% had experienced physical and sexual problems because of infertility.&#x0D; CONCLUSION: It is concluded that participants are suffered mainly from psychosocial problems beside that physical and sexual problems. Cent percent of the participants were under treatment and majority of participants said they will try for conception until fertility seized. One third of participants spouse had problem of sperm. It is recommended to concerned authority of hospital to provide couple counseling regarding infertility

Modified stepwise mini-incision microdissection testicular sperm extraction: a useful technique for patients with a history of orchidopexy affected by non-obstructive azoospermia.

Non-obstructive azoospermia (NOA), which is defined as the absence of spermatozoa in the ejaculate secondary to impaired spermatogenesis within the testis, may be caused by a variety of etiologies, including varicocele-induced testicular damage, cryptorchidism, prior testicular torsion, post-pubertal mumps orchitis, gonadotoxic effects from medications, genetic abnormalities, chemotherapy/radiation, and other unknown causes currently classified as idiopathic (Cocuzza et al., 2013). The microdissection testicular sperm extraction (micro-TESE) technique involves a meticulous microsurgical exploration of the testicular parenchyma to identify and selectively extract larger seminiferous tubules that carry a higher probability of complete spermatogenesis (Schlegel, 1999). The Cornell group evaluated the efficacy of micro-TESE in 152 NOA patients with an associated history of cryptorchidism. In their series, spermatozoa were successfully retrieved in 116/181 attempts (64%), and the resulting pregnancy rate was 50% with a delivery rate of 38% (Dabaja and Schlegel, 2013). Franco et al. (2016) described a stepwise micro-TESE approach in NOA patients, which was considered to reduce the cost, time, and effort associated with the surgery. Alrabeeah et al. (2016) further reported that a mini-incision micro-TESE, carried through a 1-cm equatorial testicular incision, can be useful for micro-TESE candidates, particularly in patients with cryptozoospermia. We conducted a retrospective study of 20 consecutive NOA patients with a history of orchidopexy from May 2015 to March 2017.

4 Sperm, but not seminal plasma, elicit changes in the bovine endometrial transcriptome after natural mating

Studies in mice and pigs have shown that seminal plasma (SP) can modulate the uterine environment, leading to an increase in embryo survival and implantation. However, in cattle, conflicting evidence has resulted from studies in which SP was infused into the uterus. In contrast to mice and pigs, it is questionable whether any SP reaches the bovine uterus during natural mating. However, sperm could act as vehicle for SP proteins that bind to their plasma membrane at ejaculation. Thus, the aim of this study was to analyse the effect of exposure to SP, in the presence or absence of sperm, on the endometrial transcriptome. Beef heifers were oestrus synchronized and mated after being seen in standing oestrus with intact bulls (which ejaculate sperm and SP, n=7) or vasectomized bulls (VX, which only ejaculate SP, n=8) or left unmated (control, n=7). To reduce a potential bull effect, 3 vasectomized and 2 intact Holstein bulls were used. Each heifer was mounted once to one intact or VX bull. Heifers were slaughtered 24h (±6h) after mating, and their reproductive tracts were recovered. Endometrial tissue from the base of the uterine horn ipsilateral to the ovulatory follicle was processed for RNA sequencing. Surprisingly, mating with a VX bull resulted in no differentially expressed genes (DEG) in the endometrium compared with unmated controls. In contrast, mating with intact bulls resulted in 22 DEG (10 down regulated and 12 upregulated) compared with unmated controls and 24 DEG (6 downregulated and 18 upregulated) compared with VX samples. The upregulated genes in intact versus VX samples included several that may have a potential role in implantation: CX3CL1, C4BPA, VCAM1-like, and IL17F. Gene CX3CL1 promotes trophoblast migration in women and has been described as a potential bovine embryokine. In addition, decreased levels of C4BPA have been correlated with implantation failure and unexplained recurrent abortions in women. Further, VCAM1 is involved in bovine conceptus implantation and is downregulated in women with unexplained infertility. In mice, SP regulates inflammation of IL17F in the uterus through the γδT/IL-17 pathway. On the other hand, the immune-related genes PRSS2 and CEBPD were downregulated in intact versus control samples, and BOLA-DQB was downregulated in intact versus VX samples. Gene PRSS2, involved in tissue remodelling, is upregulated in the endometrium of repeated breeder cows. Gene CEBPD, is a transcription factor that enhances IL6 expression, whereas BOLA-DQB is involved in antigen presentation. Interestingly, the gene coding for the α-chain of this protein is also downregulated in high fertility cows. In conclusion, these results do not support a direct effect of SP in regulating pregnancy success in cattle. However, sperm can elicit modest endometrial transcriptomic changes related to implantation and immune system regulation. Further research is needed to elucidate the role of intrinsic sperm proteins, or SP proteins transported by these cells, in the modulation of the uterine environment. This work was supported by EU Horizon 2020 Marie Sklodowska-Curie (No. 792212).

Can Testicular Size Be a Predictive Factor for Successful Sperm Retrieval in Patients with Non Obstructive Azoospermia?

Objective: Non-obstructive azoospermia, defined as absence of spermatozoa in the ejaculate caused by impaired spermatogenesis, is the most severe cause of male infertility. It is typically presented as high serum follicle stimulating hormone levels and atrophic testis. The combination of intracytoplasmic sperm injection and Microdissection testicular sperm extraction allows these infertile men the opportunity to have their own children from their own testis. Our aim was to evaluate the outcomes of micro-Testicular sperm extraction in men with atrophic testis.Study Design: The medical records of 80 non-obstructive men with azoospermia who underwent micro-TESE were retrospectively evaluated. We assessed clinical parameters; age, duration of infertility, smoking, chromosomal karyotype, Y chromosome microdeletion, follicle stimulating hormone, luteinizing hormone, total testosterone and testicular volume in relation with Microdissection testicular sperm extraction results.Results: Testicular sperm retrieval rate was 53% in 80 patients. Testicular volume, serum follicle stimulating hormone and total testosterone concentrations showed correlation with the results of sperm retrieval. These three parameters were found to be significant risk factors with testicular sperm extraction negative patients (p&lt;0.001). The odds ratios (95% CI) were 6.39 (1.25–26.58), 1.24 (1.11-1.36), 1.13 (0.99-1.21) respectively. Testicular volume was found to be a discriminative parameter in patients with negative sperm retrieval. The cut-off point was established as 6.75 ml for testicular volume with 88.1% sensitivity, 62.1% specificity.Conclusion: Microdissection testicular sperm extraction is the most effective procedure for patients with non-obstructive azoospermia. Testicular volume, serum follicle stimulating hormone and testosterone levels can be predictive factors for sperm retrieval in men with non-obstructive azoospermia.

108 Use of seminal plasma as ovulation inductor in alpacas (Vicugna pacos) embryo recipient and its effect on pregnancy rate

Alpacas, like other camelids, are classified as induced ovulators because an external stimulus is required for the occurrence of ovulation. Recent studies have identified the β-nerve growth factor (β-NGF), a protein present in the seminal plasma (SP), as responsible for inducing ovulation in this species and having potent luteotropic function. We speculate that SP can be used in reproductive biotechnologies, such as embryo transfer (ET), to increase the number of genetically superior animals produced in breeding programs. The objective of this study was to evaluate the effect of inducing ovulation with SP or a gonadotrophin-releasing hormone (GnRH) analogue on pregnancy rate in recipients of an alpaca ET program. Semen from 5 adult male alpacas was collected with an artificial vagina and diluted 1:1 (v/v) with phosphate-buffered saline (PBS; Gibco-BRL). The diluted semen was centrifuged for 30min at 1200g, and the supernatant or SP was decanted and examined for absence of sperm. Then, the SP was centrifuged again for 20min at 1200×g. The SP was stored at −80°C until the use. Female alpacas (n=38; 6-8 years) with a body condition score of 2.5-3.5 (scale 1-5) were used for the experiment. Animals were evaluated daily by transrectal ultrasonography to determine the presence of a dominant follicle ≥7mm and randomly assigned to one of two groups: (1) GnRH (n=20), 0.04mg of Buserelin acetate IM, or (2) SP (n=18): 1.0mL of seminal plasma IM. Seven days after GnRH or SP treatment (Day 0) the recipients received a fresh embryo. Alpaca donors (n=18) were given GnRH (0.04mg IM) and treated 36h later with 700IU of equine chorionic gonadotropin (ECG). Donors were mated with fertile males 5 days after ECG (Day 0 of recipients), and embryos were recovered 7 days later. Embryos of similar quality were assigned to both groups and transferred nonsurgically to the uterine horn ipsilateral to the corpus luteum. Ultrasonography examinations were performed on Day 2 to confirm ovulation and Day 25 to determine pregnancy in all of the recipients. Data was analysed by chi-squared test. Ovulation rate was not different between groups (100% each). Pregnancy rate was 45% (9 out of 20) and 44% (8 of 18) in GnRH and SP groups, respectively (P=0.77). In conclusion, SP was effective to induce ovulation in alpacas and was able to produce a pregnancy rate similar to that of GnRH as an ovulation-inducing treatment. Thus, SP can be used as an alternative for ET programs in alpacas. Research was funded by the project Role of Seminal Plasma in Reproductive Physiology and Application of Biotechnologies in Camelids (149-2017-CIENCIACTIVA).

Sperm enter glands of preovulatory bovine endometrial explants and initiate inflammation.

We previously reported that sperm binding to cultured monolayers of bovine uterine epithelial cells induces an acute inflammatory response involving the Toll-like receptor (TLR2) signaling pathway. This response serves to clear the uterus of sperm and thereby prepares the endometrium for implantation. The endometrium is lined by surface epithelial cells; however, epithelial cells also line uterine glands. To investigate the source of the immune response, we used an explant model. Explants of bovine endometrium were incubated with bull sperm illuminated by JC1 fluorescent labeling in their mitochondria. The sperm glided over the surface epithelium until they encountered and entered uterine glands, where they remained. Scanning electron microscopy of explants revealed polymorphonuclear neutrophils (PMNs) in uterine glands along with sperm. In the absence of sperm, PMNs were not seen in glands. The incubation of sperm with explants resulted in an acute inflammatory response, seen as the upregulation of mRNA expression of IL8, TNFA, IL1B, PGES and TLR2 in whole explants, as well as increased TNFA protein expression in uterine glands. TLR1/2 antagonist reduced sperm numbers in the glands and inhibited the increase of TNFA. Our observations suggest that uterine glands serve as a site where sperm interact with the uterine epithelium to trigger the innate immune response to clear excess sperm from the uterus.

Do assisted reproduction outcomes differ according to aetiology of obstructive azoospermia?

Azoospermia is defined as absence of spermatozoa and may be secondary to blocked seminal ducts, known as obstructive azoospermia. Semen quality may be impaired due to factors such as sperm cell DNA fragmentation and presence of antisperm antibodies. The objective of this article was to investigate potential differences in outcomes of in vitro fertilisation and intracytoplasmic sperm injection between groups with different obstruction aetiology, as well as between the use of different techniques and sperm cells of different origins. Retrospective, multi-centre analysis of 621 first cycles was carried out between 2008 and 2015: Group I, congenital obstruction, 45 patients and Group 2, vasectomy, 576 patients. Sperm cell retrieval was achieved in all cases. Results were similar forGroup I and II fertilisation rates, 70% versus 66.85% (p=.786); pregnancy rates, 42.5% versus 41.46% (p=.896); and live birth rates, 29.73% versus 17.69% (p=.071). According to sperm cell origin (579 epididymal vs. 42 testicular), pregnancy rates, 41.47% versus 43.9% (p=.760); andlive birth rates, 18.3% versus 27.78% (p=.163)had no difference. Fertilisation, pregnancy and live birth rates did not differ according to obstruction aetiology. Outcomes did not differ between groups according to sperm cell origin.

The appropriateness of analysing some known micro-RNAs to detect the presence of semen in old stains

Recently microRNAs (miRNAs) have gained notoriety for having revealed useful biomarkers. The advantage of using microRNAs as biomarkers lies in their ease of use and the accuracy with which they can be measured as well as their extreme tissue specificity.Furthermore, these small molecules are exceptionally stable in laboratory treatments, unlike larger RNA molecules such as messenger RNA.In the forensic field, the use of microRNAs to confirm biological evidence has become a powerful tool of crucial importance for the reconstruction of a crime scene.Semen can provide significant evidence in some types of crime, particularly sexual assault. However, a diagnosis of sperm is often difficult for several reasons, such as trace degratheir ease of use and the accuracy with which they can be measured as well as their extreme tissue specificity.Furthermore, these small molecules are exceptionally stable in laboratory treatments, unlike larger RNA molecules such as messenger RNA.In the forensic field, the use of microRNAs to confirm biological evidence has become a powerful tool of crucial importance for the reconstruction of a crime scene.Semen can provide significant evidence in some types of crime, particularly sexual assault. However, a diagnosis of sperm is often difficult for several reasons, such as trace degradation and the presence of azoospermia, which are still present when using morphological, immunological and enzymatic methods. Recent studies show that the use of micro-RNAs overcomes both the problem of degradation (due to their high resistance to exogenous factors) and the absence of spermatozoa in azoospermic individuals.In this study, some semen-specific microRNAs (miR-10a-5p, miR-888-5p, miR891a-5p, miR135a-5p, miR135b-5p) on old semen stains were tested to assess their suitability in forensic casework. The expression of this set of markers was evaluated using a miRCURY LNA Universal RT microRNA PCR System.LNA technology showed interesting properties for molecular hybridization, including enhanced specificity in allele-specific PCR and could be used to overcome the difficulties of studying very short sequences. We concluded that the cellular specificity of the studied microRNA set makes it possible to identify the presence of semen in old stains with high specificity and consequently with high probative value in a court of justice.

Management and Treatment of Varicocele in Children and Adolescents: An Endocrinologic Perspective.

Pediatric varicocele has an overall prevalence of 15%, being more frequent as puberty begins. It can damage testicular function, interfering with Sertoli cell proliferation and hormone secretion, testicular growth and spermatogenesis. Proper management has a pivotal role for future fertility preservation. The aim of this review was to discuss the diagnosis, management and treatment of childhood and adolescent varicocele from an endocrinologic perspective, illustrating the current evidence of the European Society of Pediatric Urology (ESPU), the European Association of Urology (EAU), the American Urological Association (AUA) and the American Society for Reproductive Medicine (ASRM) scientific societies. According to the ASRM/ESPU/AUA practice committee, the treatment of adolescent varicocele is indicated in the case of decreased testicular volume or sperm abnormalities, while it is contraindicated in subclinical varicocele. The recent EAS/ESPU meta-analysis reports that moderate evidence exists on the benefits of varicocele treatment in children and adolescents in terms of testicular volume and sperm concentration increase. No specific phenotype in terms of testicular volume cut-off or peak retrograde flow (PRF) is indicated. Based on current evidence, we suggest that conservative management may be suggested in patients with PRF < 30 cm/s, testicular asymmetry < 10% and no evidence of sperm and hormonal abnormalities. In patients with 10–20% testicular volume asymmetry or 30 < PRF ≤ 38 cm/s or sperm abnormalities, careful follow-up may ensue. In the case of absent catch-up growth or sperm recovery, varicocele repair should be suggested. Finally, treatment can be proposed at the initial consultation in painful varicocele, testicular volume asymmetry ≥ 20%, PRF > 38 cm/s, infertility and failure of testicular development.

Biallelic mutations in Sperm flagellum 2 cause human multiple morphological abnormalities of the sperm flagella (MMAF) phenotype.

Male patients with multiple morphological abnormalities of the sperm flagella (MMAF) are infertile and exhibit absent, short, coiled, bent and/or irregular sperm flagella. Mutations in the SPEF2 gene reduce sperm motility and cause sperm tail defects in animal models and humans. In the present study, we performed a genetic analysis on an MMAF patient and identified novel biallelic mutations in the SPEF2 gene. The biallelic mutations were confirmed by Sanger sequencing and in silico analysis revealed that, these variations were deleterious. The expression of truncated SPEF2 protein was reduced significantly in the patient's spermatozoa. The spermatozoa harbored biallelic mutations and showed severe ultrastructural defects in the axoneme and mitochondrial sheath. Our data suggest that biallelic mutations in SPEF2 can cause severe sperm flagellum defects, thus providing a novel candidate genetic pathogen for the human MMAF phenotype.

Effects of aqueous leaf extract of &lt;i&gt;Mucuna Pruriens&lt;/i&gt; (Linn.) on the weight and histology of the testis and epididymis in adult male Wistar rats

Mucuna pruriens, a leguminous plant common to Africa and Asia has been used for managing infertility in males, disorder of the nervous system, and also as an aphrodisiac. This study was aimed at investigating the effect of aqueous leaf extract of Mucuna pruriens on the weight of the animals, reproductive organs (testis and epididymis) and histology of the testis and epididymis. Twenty- five adult male Wistar rats weighing above 200g were randomized into five (5) groups consisting five (5) rats each. Group A received 1ml sterile water via oral route to compensate for stress of administration incurred in the treated groups. Group B1 and B2 received 500mg/kg body weight via oral route for 4 and 8 weeks respectively and group C1 and C2 received 1500mg/kg body weight of extract via oral route daily for 4 and 8 respectively. The extract did not cause any significant change in the body weight of the animals. A significant decrease (P&lt;0.05) was also observed in the testis of rats in groups C2 while a significant increase (P&lt;0.05) was observed in the right epididymis of rats in group B2 and C1 and left epididymis of rats in group B2. Histological findings revealed distortion of the sequential arrangement of spermatogenic cells in the seminiferous tubule epithelium, absence of spermatozoa and immature sperm cells in the lumen of the seminiferous tubule lumen. Scanty spermatozoa were observed in the epididymal lumen of rats in groups B1 and C2. Findings from this study indicated that Mucuna pruriens had an anti-fertility effect on adult male Wistar rat. Data was analyzed using SPSS/IBM statistical package version 20 and the statistical method employed was ANOVA.Key word: Mucuna pruriens, weight, testis and epididymis

A prospective double-blind, randomized, placebo-controlled study to evaluate the efficacy of silodosin 8mg as an on-demand, reversible, nonhormonal oral contraceptive for males: a pilot study.

As silodosin 8mg, used for treating BPH, is known to cause absence of sperms in the ejaculate, its potential as an on-demand, reversible, nonhormonal oral contraceptive for males was assessed in this pilot study. In this randomized, double-blind, placebo-controlled pilot study, healthy, sexually active adult male volunteers in a stable sexual relationship with a female partner with proven fertility were recruited. Those using other contraceptive methods were excluded. During the first part of the study, silodosin 8mg and placebo were crossed over among the randomly assigned two groups of participants and their post-masturbation semen and post-ejaculation urine were analyzed for spermatozoa. During the second part, both groups received silodosin 8mg 3h prior to heterosexual vaginal intercourse for 1year. The primary outcome measure was the number of unintended pregnancies in their female partners. The secondary outcome measure was dropout rate due to adverse events. The clinical trial registry number was CTRI/2017/09/009872. The study period was January 2017 to December 2017 with sample size 63. Reversible azoospermia was noted following silodosin intake during the first part of the study. During the second part, no untended pregnancy was reported. None of the participants dropped out due to adverse events. Four patients were lost to follow-up. Silodosin 8mg achieved total reversible azoospermia, hence prevented unintended pregnancies in the female partners of the males who used the drug as an on-demand oral contraceptive.

PO-01-101 Epididymovasostomy: Patency, pregnancy rate and predictive factors for success in 109 patients over a decade

This study aims to assess, and identify predictive factors for, the patency and pregnancy rates in patients with obstructive azoospermia(OA) following microsurgical epididymovasostomy (EDV). We retrospectively analysed data on 129 infertile men diagnosed with OA, who underwent EDV over a ten year period at a tertiary referral centre. Complete follow up data including semen analyses, pregnancy and patency rate was analysed in 109 patients. The patient and partner’s age, and cause of azoospermia was also noted. EDV was performed using an end to side anastomosis, with or without intussusception. Data collection on intra¬ and post-operative parameters concentrated on microsurgical anastomotic technique, evidence of spermatogenesis in aspirated epididymal fluid (none, immotile, motile), and level of epididymal anastomosis (head, body, tail). Patency rate (sperm concentration at least 0.1x106 or natural conception), pregnancy and live births rate (total number of live births) and surgeon experience was also collected. The median ages of patients and their partners were 39 ± 8.3 years and 34 ± 5.4 years respectively. The aetiology of obstruction included idiopathic (37.6%), post vasectomy (26.6%), iatrogenic (3.7%), and trauma (3.7%).Microsurgical end to side anastomosis was performed in 64.2% of cases, with 28.4% of patients undergoing end to side intussusception. The most common level of anastomoses (77%)was at the level of the epididymal body, followed by 20.2% at the level of the tail and 13.8% at the head. Intraoperative aspiration of the epididymal fluid demonstrated motile and immotile sperm in 45.9% (n=50), and 40.4% (n=44) of cases respectively, with absence of sperm in 6.4%(n=7). Patients underwent unilateral (66%) or bilateral 36.7% EDV. The majority of procedures (57.8%) were carried out by a surgeon performing >10 EDV procedures per year. There were n=6 conservatively managed complications including haematoma and infection. The patency and birth rate was 56% (n=61) and 12.5% (n=13) respectively, over a 15 month follow up period.

The Treatment of Male Infertility After Allograft Renal Transplantation: A Case Series

Background: Renal transplantation is a useful option for allowing female renal failure patients of childbearing age to achieve pregnancy. However, there have been a few reports on the effects of renal transplantation on infertility treatment in male renal failure patients. We herein report two cases in which male patients underwent infertility treatment after renal transplantation. Case Presentation: Case 1: A 51-year-old Asian (Japanese) man underwent transplantation (the donor was his wife) for renal failure due to Autosomal Dominant Polycystic Kidney Disease (ADPKD). At two years after transplantation, he visited the reproduction center in our institute due to infertility. A semen analysis revealed oligoasthenozoospermia. He ultimately failed to achieve pregnancy and gave up on infertility treatment. Case 2: A 47-year-old Asian (Japanese) man underwent renal transplantation (the donor was his sister) due to renal failure caused by diabetes mellitus. At three years after renal transplantation, he visited the reproduction center in our institute for infertility. Due to ejaculation disability and the absence of sperm in the patient’s urine after masturbation, he was diagnosed with anejaculation. Thus, testicular sperm extraction (TESE) was performed. Twenty-three motile spermatozoa were successfully retrieved by microdissection TESE (micro-TESE). ICSI was subsequently performed and a good embryo was transferred. His wife achieved pregnancy and is expected to deliver this October. Conclusion: We report two cases of male infertility treatment after renal transplantation.

99 Effect of application of seminal plasma on Day 0, 5, or 7 postmating on pregnancy rate and embryonic survival in alpacas (Lama pacos)

Alpacas, similar to other camelids, are classified as induced ovulators, as an external stimulus is required for the occurrence of ovulation. A protein identified as β-nerve growth factor, present in the seminal plasma (SP), has the capacity to induce ovulation and corpus luteum formation. Alpacas exhibit poor reproductive efficiency, with birth rates below 50% due in part to high embryo mortality before 35 days post-mating. A study was carried out to evaluate the effect of the application of SP on Day 0, 5, or 7 post-mating on pregnancy rate and embryo survival, defined as the difference in the numbers of pregnant females between Day 35 and 25. Nonpregnant adult alpaca females (n=124) between 5 to 6 years old were evaluated by transrectal ultrasonography to determine presence of a follicle ≥7mm, and then 2 days later to confirm permanence of the follicle. Alpacas were then bred by natural mating and assigned randomly to 1 of 4 treatments: 1mL of SP IM at mating; 1mL of SP IM Day 5 post-mating; 1mL of SP IM Day 7 post-mating; and control. Semen was collected from adults male and ejaculates were diluted 1:1 with PBS and then centrifuged for 30min at 3000 rpm. Supernatant was separated and a drop evaluated to determine absence of spermatozoa and SP-free sperm was stored at −20°C. Twenty adult males with optimal reproductive performance were used for mating with females assigned to the different treatments. Animals were evaluated by ultrasound with an Aloka SSD 500 (Aloka, Tokyo, Japan) and 5.0-MHz linear transducer on Day 25 and 35 to determine pregnancy rate and embryonic survival. Data were analysed by chi-square. Results are present in Table 1. The results differ from our initial hypothesis and a possible explanations may be that additional application of SP IM could saturate receptors and block the action of the seminal plasma present in the ejaculate of males. Table 1.Pregnancy rate and embryonic survival in alpacas with application of seminal plasma on Day 0, 5, or 7 Study was supported by project no. 405-PNICP-PIAP-UNMSM.

A novel stopgain mutation c.G992A (p.W331X) in TACR3 gene was identified in nonobstructive azoospermia by targeted next-generation sequencing.

Nonobstructive azoospermia (NOA) is one of the most severe forms of male infertility because of impaired spermatogenesis with the absence of spermatozoa in the ejaculate. The causes of this disease can be partly attributed to genetic factors. Some common structural variants and single nucleotide polymorphisms (SNPs) were reported to be associated with NOA. However, the underlying etiology and genetic mechanism(s) remain largely unclear. The aim of this study was to investigate the associated mutations of spermatogenic genes in Chinese infertile men with NOA. The entire coding region of 25 genes associated with spermatogenesis was sequenced from 200 infertile men with NOA. Screening was carried out using the targeted exome sequencing to identify genetic variations and SNPs of the entire coding region of these genes. After the targeted exome sequencing data were filtered through several currently existing variation databases, a series of variations were found. In this paper, we report one novel stopgain variation c.G992A (p.W331X) in the exon 4 of TACR3 gene. The variant was heterozygous and categorized as pathogenic. In conclusion, our study revealed a novel stopgain mutation c.G992A (p.W331X) in TACR3 which expanded the mutation spectrum of TACR3 in Chinese NOA infertile men and advanced our understanding of the genetic susceptibility to NOA.