Obstructive Azoospermic Men Research Articles

The MAEL expression in mitochondria of human spermatozoa and the association with asthenozoospermia.

The maelstrom spermatogenic transposon silencer (MAEL) function in postmeiotic germ cells remains unclear, and its protein localization in human testis and spermatozoa awaits determination. This study aims to clarify the MAEL expression in human spermatogenesis and to explore its role in sperm function. Twenty-seven asthenozoospermic men, 40 normozoospermic controls, and three obstructive azoospermic men were enrolled. The transcripts of MAEL in the seminiferous epithelium and MAEL downstream targets were identified by bioinformatics analysis. MAEL protein expression in human testis and ejaculated sperms were examined by immunohistochemical and immunogold staining, respectively. The roles of MAEL in mitochondria function were investigated by siRNA knockdown in human H358 cells. The association between MAEL protein levels and clinical sperm features was evaluated. Abundant MAEL was expressed in spermatid and spermatozoa of the human testis. Remarkably, MAEL was located in the mitochondria of ejaculated sperm, and bioinformatics analysis identified GPX4 and UBL4B as MAEL's downstream targets. Knockdown of MAEL sabotaged mitochondria function and reduced adenosine triphosphate (ATP) production in H358 cells. MAEL, GPX4, and UBL4B expression levels were significantly decreased in asthenozoospermic sperms than in controls. The MAEL protein levels were positively correlated with GPX4 and UBL4B in human sperm. Total motile sperm count (TMSC) was positively correlated with protein levels of MAEL, GPX4, and UBL4B in ejaculated sperms. We highlight prominent MAEL expression in the intratesticular spermatid and the mitochondria of ejaculated spermatozoa. MAEL directly binds to GPX4 and UBL4B, and loss of MAEL induces mitochondrial dysfunction. MAEL-mitochondrial function-motility relationship might advance our understanding of the causes of asthenozoospermia.

Factors predicting the outcome of percutaneous epididymal sperm aspiration in men with obstructive azoospermia

To report on the live birth rates (LBRs) following percutaneous epididymal sperm aspiration (PESA) in men with obstructive azoospermia (OA) and factors affecting treatment outcome which is under reported in the literature. This is a multicenter study that was conducted in Egypt including all couples undergoing intra cytoplasmic sperm injection (ICSI) for OA using PESA-derived sperms. Men were subdivided according to aetiology into congenital, iatrogenic and idiopathic groups. Fertilization, pregnancy and LBRs were determined and compared in each group. The longitudinal LBR, crude and expected cumulative delivery rates (CCDR, ECDR) were calculated. Multiple logistic regression analysis was used to determine significant associations between maternal, paternal and ICSI factors with successful live births. Ninety couples were included in the study. Viable sperm for ICSI was retrieved in 89 men (98.9%). A total of 155 ICSI cycles with 17 frozen embryo transfers resulted in 81 pregnan-cies and 55 live births. After 5 cycles, the longitudinal LBR, CCDR and ECDR were 30%, 57.3% and 88.6% respectively. Maternal age and number of fertilized eggs were the only fac-tors significantly affecting LBRs. PESA is a minimally invasive procedure for secur-ing viable sperm for ICSI in OA men, with high cumulative delivery rates. Maternal age and number of fertilized eggs are the only factors that significantly affecting LBR. The contempo-rary longitudinal and cumulative LBRs provide objective out-come data to counsel OA patients undergoing fertility treat-ments.

445 Relevance of testicular histopathology on prediction of sperm retrieval rates after a conventional bilateral multiple TESE in case of non-obstructive azoospermia (NOA) and obstructive-azoospermia (OA)

To establish the relevance of testicular histopathology (TH) on sperm retrieval (SR) after testicular sperm extraction (TESE) in patients with non-obstructive azoospermia (NOA) and in obstructive azoospermia (OA) patients, already underwent a previous failure testicular fine needle aspiration (TEFNA). We evaluated a total of 82 azoospermic men, underwent TESE, referring to our Centre 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 (FSH), luteinizing hormone (LH) and total testosterone (TT) were collected. Successful sperm retrieval (SSR) was obtained in 36 men of total (43.9%).(Table 1) SSR was 29.5% in NOA patients, while OA men, underwent a previous failure TEFNA, had SR in 86% of cases. Mean LH was 6.55 IU/L, TT 4.70 ng/ml, testicular volume (TV) right 13.7 ml and TV left 13.6 ml. FSH mean was 13.45 IU/L in patients with negative SR and 8.18 IU/L in men with SSR. According to histology, 20.7% had Normal Spermatogenesis (NS), 35.3% Hypospermatogenesis (H), 35.3% Maturation Arrest (MA) and 8.5% Sertoli-Cell-Only-Syndrome (SCOS). SSR was 88.2% in patients with NS, 24.1% in the MA group and 48.27% in patients with Hypospermatogenesis, while negative SR was reported in SCOS patients. 2 patients with NS and unsuccessful SR, underwent a second TESE, of whom one had SR and cryopreservation.

Effect of maca(Lepidium meyenii) aqueous extract on the epididymal sperms quality and the DNA normality of vasectomized mature mice:model forobstructive azoospermia in men

This study was aimed to found out the effect of Maca extract on certain sperm function characters and sperm DNA normality in vasectomized and healthy adult mice as a model for men complaining from obstructive azoospermia. Twenty adult male mice were randomly divided into four groups (5males per each group).The first group(GI) was regarded as a negative control that treated with distilled water. Whereas group II was gavage maca extract daily with 1mg/100gm.GropIII,and IV were vasectomized . Mice in GIII were vasectomized without treatment while the mice in group GIV were vasectomized and orally administrated maca dose(1mg/100gm).At the last of experiment(35 days), all the mice were scarified for assessment of certain sperm function parameters . The results revealed that a significant (P<0.05)improvement of grade (A and B) of active sperm motility in mice treated orally with Maca extract was recorded compared with other groups, while grade (C) was significantly increase in vasectomized mice (GIIIgroup) as compared with other healthy mice . DNA fragmentation resulted from fertile mice gavages with maca were reduced compared to healthy non-treated group and other treated and non-treated vasectomized mice (GIII and GIVgroup).Orally administration of animals with Maca extract caused a significant (p<0.05) improvement in the percentage of morphologically normal sperm compared with vasectomized treated and non treated groups.
 It is concluded that oral administration of Maca extract caused a significant increment in certain sperm function parameters of vasectomized mice in turn this result can be utilized for obstructive azoospermic men.

Effect of Fertility Blend® Administration on the Epididymal Sperm Function Parameters of Vasectomized Mice: Physiological and Genetical Study as Model for Obstructive Azoospermic Men


 
 
 
 Male Fertility Blend® is a new nutrient supplements containing many constituents especially a plant called Dong quai(Angelia Sinensis). The plant extract has been used to facilitate the sperm function parameters. However, the studies concern on its effects on epididymal sperms of vasectomized males and obstructive azoospermia are very rare. Thus, this investigation was designed to elucidate the role of Male Fertility Blend® (MFB) formula on the in vivo epididymal sperm characters and DNA fragmentations of vasectomized male mice as a model for man. In this study, the orally administration of 3.4 µg/ml MFB was used for vasectomized and non-vasectomized mice along 35 days. The results revealed a significant (p≤0.05) increment in certain sperm function parameters of vasectomized mice by using fertility Blend® than that of not treated by this supplement. The percentage of progressive and unprogressive active sperm motility using MFB was significantly (P≤0.05) increased compared with non-treated group. It was concluded that the MFB formula containing different sources of energy and variety of factors that sustain the epididymal sperm of healthy and vasectomized mice. Therefore this supplement can be utilized for males complaining from obstructive azoospermia and other factors of infertility.
 
 
 

Testicular sperm correlates with a similar rate of euploid embryo development compared to cycles using ejaculated sperm

To evaluate the percentage of euploid embryo development in patients with obstructive and non-obstructive azoospermia. Retrospective data analysis. From March 2013 to April 2016, 36 azoospermic men underwent 45 cycles of ICSI-PGS cycles with blastocyst trophoectoderm biopsy for comprehensive chromosomal screening using high density oligonucleotide microarray comparative genomic hybridization. 30 cycles used testicular sperm from obstructive azoospermia men with prior vasectomy, 9 cycles were non-obstructive, and 6 cycles were from azoospermic men with unknown causes. The results were compared with 621 In-Vitro Fertilization (IVF) patients that used ejaculated sperm during the same time period. 3,475 blastocysts were generated and underwent trophoectoderm biopsy and PGS. Tabled 1Age group# ICSI Cycle# MII eggs ICSI’d#2PN(Fert rate)Blastocyst # (rate)Euploid #(rate)P value<37TESE38446341 (76.5%)206 (60.4%)133/163 (81.6%)P=0.6<37EJ40143723096 (70.8%)3271/5361 (61.0%)1911/2491 (76.7%)38-42TESE75448 (88.5%)19 (39.6%)10/19 (52.5%)P=0.438-42EJ2201044885/1044 (84.8%)771/1537 (50.2%)306/802 (38.2%) Open table in a new tab Our data demonstrate that testicular sperm from azoospermic patients have comparable fertilization, blastocyst formation and euploid rates compared to cycles using ejaculated sperm. These comparable rates of fertilization, blastocyst formation and rates of euploid embryos with the use of testicular sperm requires further study but may be due in part to female partners with more favorable fertility potential.

Altered Crossover Distribution and Frequency in Spermatocytes of Infertile Men with Azoospermia.

During meiosis, homologous chromosomes pair to facilitate the exchange of DNA at crossover sites along the chromosomes. The frequency and distribution of crossover formation are tightly regulated to ensure the proper progression of meiosis. Using immunofluorescence techniques, our group and others have studied the meiotic proteins in spermatocytes of infertile men, showing that this population displays a reduced frequency of crossovers compared to fertile men. An insufficient number of crossovers is thought to promote chromosome missegregation, in which case the faulty cell may face meiotic arrest or contribute to the production of aneuploid sperm. Increasing evidence in model organisms has suggested that the distribution of crossovers may also be important for proper chromosome segregation. In normal males, crossovers are shown to be rare near centromeres and telomeres, while frequent in subtelomeric regions. Our study aims to characterize the crossover distribution in infertile men with non-obstructive (NOA) and obstructive azoospermia (OA) along chromosomes 13, 18 and 21. Eight of the 16 NOA men and five of the 21 OA men in our study displayed reduced crossover frequency compared to control fertile men. Seven NOA men and nine OA men showed altered crossover distributions on at least one of the chromosome arms studied compared to controls. We found that although both NOA and OA men displayed altered crossover distributions, NOA men may be at a higher risk of suffering both altered crossover frequencies and distributions compared to OA men. Our data also suggests that infertile men display an increase in crossover formation in regions where they are normally inhibited, specifically near centromeres and telomeres. Finally, we demonstrated a decrease in crossovers near subtelomeres, as well as increased average crossover distance to telomeres in infertile men. As telomere-guided mechanisms are speculated to play a role in crossover formation in subtelomeres, future studies linking crossover distribution with telomere integrity and sperm aneuploidy may provide new insight into the mechanisms underlying male infertility.

Differential expression of microRNAs and their messengerRNA targets in men with normal spermatogenesis versus Sertoli cell-only syndrome

ObjectiveTo identify the profiling of testicular microRNAs (miRNAs) that are differentially expressed in men with normal spermatogenesis versus Sertoli cell-only syndrome (SCOS), to predict the miRNA target genes, and to determine the molecular networks/pathways constituted by miRNA targets. Materials and methodsNine obstructive azoospermic men with normal spermatogenesis and nine SCOS men were enrolled in this study. Testicular tissues from three men with normal spermatogenesis and three men with SCOS were pooled respectively for miRNA microarray analysis, and the other 12 testicular specimens were used for subsequent quantitative real-time polymerase chain reaction analysis for validation. Moreover, the predicted targets of the differentially expressed miRNAs were identified in silico and uploaded to Ingenuity Pathway Analysis for further network/pathway analysis. ResultsThree miRNAs that were upregulated (miR-136, miR-630, and miR-663) and seven miRNAs that were downregulated (miR-15b, miR-18a, miR-25, miR-30a-5p, miR-34b, miR-93, and miR-126) in SCOS specimens were identified. A total of 51 spermatogenesis-related targets were predicted in silico. Results from quantitative reverse transcription polymerase chain reaction generally correlate with microarray and computational analysis. Three significant molecular networks and five canonical pathways associated with biological functions of steroidogenesis and spermatogenesis were identified. ConclusionsAberrant expression of 10 differentially expressed miRNAs might result in dysregulation of steroidogenesis and spermatogenesis that may ultimately lead to SCOS.

Multiple needle‐pass percutaneous testicular sperm aspiration as first‐line treatment in azoospermic men

Percutaneous testicular sperm aspiration (TESA) has been known for decades as a simple, minimally invasive approach to sperm retrieval in azoospermic men. Because of lower reported sperm retrieval rates (SRR) when compared with microdissection testicular sperm extraction (mTESE), many centers now use mTESE as the first choice for retrieving spermatozoa in nonobstructive azoospermia (NOA). Objectives of this study were to evaluate the outcome and safety of TESA and mTESE in the treatment of azoospermia and to investigate the usefulness of a prognostic TESA to individualize protocols for couples and limit the use of invasive testicular procedures. IRB approval was obtained to retrospectively evaluate 208 patients undergoing multiple needle-pass TESA between 1999 and 2014. Prognostic TESA was performed on 125 men with NOA and 82 with obstructive azoospermia (OA). Nine NOA men and 31 OA men with previously demonstrated spermatozoa had a subsequent therapeutic TESA while nine NOA men with a failed TESA proceeded to mTESE. Main outcome measures were complication rates and SRR. SRR of the prognostic TESA was 30% (38/125) for NOA men and 100% (82/82) for OA men. Eight/nine NOA men and 31/31 OA men had spermatozoa found for intracytoplasmic sperm injection in a subsequent therapeutic TESA. In nine NOA men in whom a TESA produced no spermatozoa, only one had spermatozoa found with mTESE. Overall complication rates of TESA and mTESE were 3% (7/267) and 21% (3/14), respectively. TESA provides reasonable SRR and is a safe procedure. Successful prognostic TESA indicates future success with therapeutic TESA. Men with a failed TESA have a limited chance of sperm retrieval using mTESE. Approaching azoospermic men with an initial prognostic TESA followed by either therapeutic TESA and/or mTESE is an efficient algorithm in the management of azoospermia and limits the use of more invasive procedures.

Antisperm protein targets in azoospermia men.

BACKGROUND:The number of couples that meet the definition of infertility at reproductive ages is increasing worldwide. One of the most known conditions of infertility in males is azoospermia, defined as complete absence of spermatozoa in the semen. Azoospermia manifests in two forms, namely obstructive and non-obstructive azoospermia. Although the presence of antisperm antibody (ASA) has been reported in 88% of the patients with obstructive azoospermia (OA), interestingly, there is no data regarding ASA targets in OA individuals.AIM:The present study aimed to identify sperm antibody targets in a group of OA men.SETTINGS AND DESIGN:The present study was carried out on 27 OA infertile men and 27 healthy fertile age-matched males as cases and controls, respectively.SUBJECTS AND METHODS:The sperm proteome was separated using two-dimensional gel electrophoresis technique, transferred onto the polyvinylidene fluoride membrane, and blotted with the sera of a group of OA men. Then, it was compared with the membranes blotted with the sera of a group of healthy fertile men. Matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI TOF/TOF) mass spectrometry was used to identify the different blotted spots and finally the results of the mass analysis were confirmed using reverse transcriptase polymerase chain reaction method.RESULTS:The results indicated that OA patients might produce antibody against two sperm proteins, Tektin-2 and triose phosphate isomerase. Moreover, the expressions of the two targeted proteins were confirmed at RNA level.CONCLUSIONS:The findings of the present study revealed two functionally important sperm proteins as antibody targets in azoospermic men.

Correlation between leptin content and sperm retrieval in cases of functional azoospermia

IntroductionTesticular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI) became the preferable techniques for solving the problem of azoospermic men. Non-invasive techniques are needed to predict sperm retrieval chance before TESE to avoid the psychological and physiological problems that may be developed. AimTo investigate the correlation between serum, seminal and testicular leptin levels and sperm retrieval in functional azoospermic men. MethodsThe study included 61 men classified into 4 groups; normozoospermia (NOR), obstructive azoospermia (OA), positive non-obstructive azoospermia (NOA (+)) and negative non-obstructive azoospermia (NOA (−)). Blood FSH, LH, Prolactin, Free and Total testosterone levels plus serum and seminal leptin levels were measured for all groups. For azoospermic groups, TESE and testicular leptin level were applied. Main outcome measuresBoth OA and NOR groups were used as control groups. The prediction accuracy for FSH and serum, seminal and testicular leptin was compared by receiver operating characteristic (ROC) curve analysis. ResultsThere were no significant differences in serum leptin levels among the four groups. Azoospermic groups showed higher seminal leptin levels than the NOR group. Seminal and Testicular leptin levels of NOA (−) men were significantly increased in comparison with OA and NOA (+) men. There was a significant negative correlation between serum leptin and total testosterone concentrations, and a significant positive correlation between testicular and seminal leptin concentrations. In ROC curve; for differentiation between positive and negative NOA, areas under the curve (AUC) of testicular and seminal leptin were greater than that of serum leptin. The combination of seminal leptin with FSH gave AUC greater than that of FSH alone. ConclusionThere is a role for leptin in spermatogenesis, and seminal leptin can be used as a good assistant marker to increase the prediction accuracy for sperm retrieval in NOA men especially in combination with FSH.

The relationship between anogenital distance and azoospermia in adult men

Anogenital distance (AGD) is a marker for endocrine disruption in animal studies in which decreased male AGD has been associated with testicular dysfunction. The objective of the study was to investigate whether anogenital distance could distinguish men with obstructive azoospermia (OA) from those with nonobstructive azoospermia (NOA). To accomplish this, azoospermic men were recruited and evaluated at a men's reproductive health clinic in Houston, TX. Anogenital distance (the distance from the posterior aspect of the scrotum to the anal verge) and penile length (PL) were measured using digital calipers. Testis size was estimated by physical examination. Logistic regression was used to compare AGD lengths in men with OA and men with NOA. A total of 69 OA men (mean age: 44.2 ± 9.2) and 29 NOA men (mean age: 32.8 ± 4.8) were recruited. The NOA men possessed significantly shorter mean AGD than the men with OA (AGD: 36.3 vs. 41.9 mm, p = 0.01). An AGD of less than 30 mm, had a 91% specificity in accurately classifying NOA. Moreover, after adjustment for age, race, and BMI, an AGD of less than 30 mm yielded a significantly increased odds of NOA compared to OA (OR 5.6, 95% CI 1.0, 30.7). In summary, AGD may provide a novel metric for assessing testicular function in men and in distinguishing OA from NOA.

Identification and characterization of repopulating spermatogonial stem cells from the adult human testis

This study was conducted to identify and characterize repopulating spermatogonial stem cells (SSCs) in the adult human testes. Testes biopsies from obstructive azoospermic patients and normal segments of human testicular tissue were used. Flow cytometry, real-time PCR and immunohistochemical analysis were performed. Purified human spermatogonia were transplanted into busulfan-treated recipient mouse testes and integrated cells were detected by human nuclear protein antibody co-localized with stem cell and germ cell markers. Testicular biopsies collected from obstructive azoospermic men showed similar morphology and distribution of markers to the normal human testes. Flow cytometry showed distinct populations of stage-specific embryonic antigen-4 (SSEA-4), CD49f and CD90 positive cells in the adult human testes. SSEA-4 (+) cells showed high expression levels of SSC-specific genes and high levels of telomerase activity. Extensive colonization of human cells in the mouse testes indicates the presence of highly enriched populations of SSCs in the SSEA-4 (+) sorted cells. All the HNP (+) cells in the mouse testes were positive for germ cell marker dead box mRNA helicase and only half of them were dimly positive for c-kit. In addition, subpopulations of human spermatogonia that colonized mouse testes were positively stained for CD49f, GPR-125, Nanog and Oct-4 indicating the existence of population of cells among human spermatogonia with SSC and pluripotent characteristics. This study clearly demonstrates that repopulating human SSCs have phenotypic characteristics of SSEA-4(+), CD49f(+), GPR-125(+)and c-Kit (neg/low). The results have direct implications for enrichment of human spermatogonia for further culture and germ cell differentiation studies.

The TNP1 haplotype - GCG is associated with azoospermia

Transition nuclear proteins (TNP1 and TNP2) are the major nuclear proteins that replace somatic histones during spermatogenesis. TNPs are required for the maintenance of normal spermatogenesis. Moreover, spermatogenesis was found to be compromised in both Tnp1 and Tnp2 null mice. As no study is available on the role of these genes in Indian infertile men, we have sequenced the entire TNP1 and TNP2 genes in 320 infertile men and 280 control fertile men drawn from two states in India. We identified 18 variants, including 8 previously known and 10 novel. Of the 10 novel variants, 3 were found only in azoospermic men, of which 2 (g.-688A>T in TNP1 and g.1030G>A in TNP2) were predicted to affect the transcription factor binding sites and therefore can cause deregulation of gene expression. Haplotype association analysis showed a significant omnibus association (omnibus χ(2) = 7.87, p = 0.0195) for the single nucleotide polymorphisms (SNPs) in the TNP1 gene with azoospermia. The frequency of the haplotype GCG (H3) was increased in azoospermic men (53.1%) compared with fertile men (43%; χ(2) = 7.964, p = 0.005). However, similar analysis of the TNP2 gene did not show any association with infertility. Furthermore, expression analysis of the TNP1 gene in obstructive azoospermic men showed that haplotypes of the TNP1 gene do not affect its expression level. Our results suggest that the individual SNPs of the TNP1 and TNP2 genes are not associated with infertility; however, the haplotype GCG of the TNP1 gene is a risk factor for azoospermia.

Differentiation of human round spermatids into motile spermatozoa through in vitro coculture with Vero cells.

This study was undertaken to examine whether human early round spermatids will differentiate in an in vitro coculture with Vero cells. A total of 1450 and 400 isolated early round spermatids mechanically collected from two non-obstructive and three obstructive azoospermic men with a normal karyotype were cocultured on Vero cell monolayers in minimum essential medium plus 10% fetal bovine serum, with or without 50 or 100 IU/L FSH and 1 or 10 μmol/L testosterone, at 32.5°C, in an environment of 5% CO2 in air. Morphological changes of the spermatids were observed microscopically. After 7 days of coculture, almost half (40-50%) of the round spermatids from both non-obstructive and obstructive azoospermic men resumed spermiogenesis in vitro. Only cells from the latter patients gave rise to spermatozoa, a few of which had a motile flagellum. Low concentrations of FSH and testosterone increased the percentage of in vitro spermiogenesis. Isolated round spermatids can resume spermiogenesis in vitro when cocultured on a Vero cell monolayer.

How successful is repeat testicular sperm extraction in patients with azoospermia?

Little is known about the extraction rate in repeated sperm retrieval procedures in azoospermic patients. This study aimed to assess the feasibility of repeated sperm recovery in these patients. A total of 1066 azoospermic men had their first sperm recovery between 1 January 1995 and 31 December 2003. A total of 381 men had obstructive azoospermia (OA), 628 nonobstructive azoospermia (NOA) and 57 showed hypospermatogenesis. Overall, sperm could be retrieved in all procedures in the 598 cycles performed in OA men (100%). A total of 117, 57, 24, 11, 7 and 1 men underwent, respectively, two, three, four, five, six and seven sperm retrievals; all were successful. Of the 784 procedures performed on the 628 men with NOA, sperm could be retrieved in 384 procedures (49%). During the first testicular sperm extraction (TESE) procedure, sperm could be extracted in 261 men with NOA (41.6%). A total of 103 men had a second attempt, 34 had a third attempt, 11 had a fourth attempt, 6 had a fifth attempt and 2 had a sixth attempt. In these cycles, sperm could be extracted in, respectively, 77 (74.7%), 28 (82.3%), 11 (100%), 5 (83.3%) and 2 (100%) men. Repeated TESE ensures a high sperm recovery rate even in patients with NOA. In NOA patients, studies reporting on TESE may therefore overestimate the retrieval rate by reallocating successful patients. These data also show that when no spermatozoa can be obtained after thawing cryopreserved testicular sperm for ICSI in NOA patients, a repeat TESE procedure can be planned.

1501: High Prevalence of Testicular Tumors in Non Obstructive Azoospermic Men

1501: High Prevalence of Testicular Tumors in Non Obstructive Azoospermic Men

Diagnostic and Prognostic Value of Testicular Fine Needle Aspiration in Non-Obstructive Azoospermia

Modern techniques of assisted reproduction may be useful also in non obstructive azoospermic men using sperm extracted from the testis (TESE). However, no clinical or hormonal parameter has reliable prognostic value in predicting the presence of testicular sperm. We report here our experience with fine needle aspiration cytology (FNAC) of the testes used as a diagnostic tool in non obstructive azoospermic patients looking at its possible role as prognostic parameter for the presence of testicular sperm at TESE. Methods One hundred and twenty-five non obstructive azoospermic men candidates to ICSI were analysed for FSH, LH, testosterone, and inhibin B plasma levels, and classified in three groups on the basis of FNAC: Sertoli cell-only syndrome (70), severe hypospermatogenesis (42), and maturation arrest (13). After that, all men underwent TESE for sperm recovery for ICSI. Results FNAC detected sperm in 24 of 42 men with severe hypospermatogenesis and 9 of 13 men with maturation arrest. TESE retrieved sperm in all such patients, as well as in additional 29 of 70 men with Sertoli cell-only syndrome, additional 11 cases of severe hypospermatogenesis (35 of 42 in total) and 1 additional case of maturation arrest (10 of 13 in total). Testicular volume and hormonal parameters were not different in men with and without sperm retrieved. Conclusion These findings suggest that FNAC may be a simple and valid diagnostic parameter in non obstructive azoospermic men, and it may represent a valid positive prognostic parameter for sperm recovery at TESE.

Should cryopreserved epididymal or testicular sperm be recovered from obstructive azoospermic men for ICSI?

To determine the effect of the anatomical site of sperm recovery on intracytoplasmic sperm injection (ICSI) embryo implantation, pregnancy and live birth rates in couples with isolated obstructive azoospermia as the sole cause of infertility. Controlled, single centre, retrospective clinical study. University Hospital, Centre for Reproductive Medicine. One hundred and fifty-one cycles of ICSI were performed, using surgically recovered sperm, between August 1996 and March 2002. The outcome of ICSI, with surgically recovered sperm, was compared between epididymal (Group E) and testicular (Group T) derived sperm. Inclusion was limited to couples undergoing their first treatment cycle, where female age was < or =39 years and a minimum of five oocytes were available for injection. Women with a history of ovarian surgery, ultrasonic evidence of polycystic ovaries, uterine anomalies or hydrosalpinx were excluded. Clinical pregnancy, implantation and live birth rate. Forty-two of 151 cycles met the strict inclusion criteria. Groups E and T were comparable with respect to age, basal serum FSH, ovarian response; number of oocytes injected and number of embryos available and transferred. No difference existed between Groups E and T in implantation, clinical pregnancy or live birth rate (28.8% vs 25.8%, 42.9% vs 42.9% and 39.3% vs 42.9%, respectively). Cryopreserved epididymal and testicular sperm, from men with obstructive azoospermia, appear equally effective in ICSI. Epididymal recovery should remain the method of first choice for obstructive azoospermic men but further study of sperm DNA damage rates in different testicular sites is required.

Chromatin condensation and DNA damage of human epididymal spermatozoa in obstructive azoospermia

Obstructive azoospermia (OA) includes patients with different aetiologies of obstruction such as congenital bilateral absence of vas deferens (CBAVD), post-vasectomy/failed vaso–vasostomy, or unknown (idiopathic). The frequency of DNA-damaged spermatozoa and abnormal chromatin condensation in epididymal spermatozoa of OA patients was investigated for the different aetiology subgroups. DNA damage in caput epididymis spermatozoa was assessed with the TdT-UTP nick-end labelling (TUNEL) assay and chromatin condensation status was measured with chromomycin A3 (CMA3) stain ( n = 60 patients). All epididymal sperm samples showed high levels of TUNEL positivity (mean and SD, 43 ± 17%). CMA3 rates showed large variation within the samples (mean and SD, 53 ± 21%, range 7–97%); however, a significant difference in chromatin condensation was found between the different aetiologies of obstruction. The highest percentage of CMA3 positivity was found in the CBAVD and idiopathic groups (60 ± 17 and 65 ± 18% respectively) compared with the post-vasectomy samples (37 ± 17%) or ejaculated spermatozoa from normozoospermic males (29 ± 12%). In conclusion, despite the normal testicular histology in OA men, chromatin condensation differed between OA patients. The physiological heterogeneity found in chromatin condensation between patient subgroups is probably influenced by the aetiology of the obstruction.