Non-obstructive Azoospermia Group Research Articles

Seminal plasma anti-M�llerian hormone level correlates with semen parameters but does not predict success of testicular sperm extraction (TESE)

To assess seminal plasma anti-Müllerian hormone (AMH) level relationships in fertile and infertile males. Eighty-four male cases were studied and divided into four groups: fertile normozoospermia (n = 16), oligoasthenoteratozoospermia (n = 15), obstructive azoospermia (OA) (n = 13) and non-obstructive azoospermia (NOA) (n = 40). Conventional semen analysis was done for all cases. Testicular biopsy was done with histopathology and fresh tissue examination for testicular sperm extraction (TESE) in NOA cases. NOA group was subdivided according to TESE results into unsuccessful TESE (n = 19) and successful TESE (n = 21). Seminal plasma AMH was estimated by enzyme linked immunosorbent assay (ELISA) and serum follicular stimulating hormone (FSH) was estimated in NOA cases only by radioimmunoassay (RIA). Mean seminal AMH was significantly higher in fertile group than in oligoasthenoteratozoospermia with significance (41.5 +/- 10.9 pmol/L vs. 30.5 +/- 10.3 pmol/L, P < 0.05). Seminal AMH was not detected in any OA patients. Seminal AMH was correlated positively with testicular volume (r = 0.329, P = 0.005), sperm count (r = 0.483, P = 0.007), sperm motility percent (r = 0.419, P = 0.021) and negatively with sperm abnormal forms percent (r = -0.413, P = 0.023). Nonsignificant correlation was evident with age (r = -0.155, P = 0.414) and plasma FSH (r = -0.014, P = 0.943). In NOA cases, seminal AMH was detectable in 23/40 cases, 14 of them were successful TESE (57.5%) and was undetectable in 17/40 cases, 10 of them were unsuccessful TESE (58.2%). Seminal plasma AMH is an absolute testicular marker being absent in all OA cases. However, seminal AMH has a poor predictability for successful testicular sperm retrieval in NOA cases.

370 BILATERAL DIFFERENTIAL TESTICULAR BIOPSIES IMPROVE TESE AND ICSI OUTCOMES IN NON-OBSTRUCTIVE AND OBSTRUCTIVE AZOOSPERMIC INFERTILE PATIENTS

The goal of the present study was to evaluate the clinical significance of bilateral differential testicular biopsies (BDTB) to improve testicular sperm extraction (TESE) and ICSI outcomes in non-obstructive azoospermic (NOA) men. The male patients were divided into an obstructive azoospermic group (OAG, n = 40) and an NOA group (n = 50). The women in both groups had normal ovulatory cycles and reproductive hormone concentrations. BDTB were taken from upper, middle, and lower zones of the testes. At least 50 transverse histologic sections of seminiferous tubules were examined in each biopsy. The FSH, LH, and prolactin concentrations were significantly higher in the NOA group compared to the OAG (18.3 vs. 6.2 mIU mL-1, 8.5 vs. 4.1 mIU mL-1, and 9.6 vs. 6.4 ng mL-1, respectively; P &amp;lt; 0.01), whereas the testosterone level and testicular size were significantly lower in the NOA group vs. the OAG (3.7 vs. 5.3 ng mL-1 and 3.3 vs. 4.4 cm, respectively; P &amp;lt; 0.01). A new method (BDTB) for scoring testicular biopsies was used. This method is different from Johnson&amp;apos;s method as SCO or TF or complete MA. Following positive BDTB results, TESE and ICSI were performed. The oocyte recovery rate/patient was 3.8&amp;percnt; (154/40) and 3.6&amp;percnt; (178/50) in the OAG and the NOA group, respectively (P &amp;gt; 0.05). The TESE-ICSI rates were 75.9&amp;percnt; (117/154) in the OAG and 59.5&amp;percnt; (106/178) in the NOA group (P &amp;lt; 0.005). The embryo cleavage rate was significantly higher in the OAG compared to the NOA group (98/117 &amp;equals; 83.8&amp;percnt; vs. 66/106 &amp;equals; 62.3&amp;percnt;, respectively; P &amp;lt; 0.005). The number of embryos transferred/patient in the OAG was significantly higher compared to that in the NOA group (2.5 vs. 1.3, respectively; P &amp;lt; 0.01). The embryo implantation rate per embryo transfer in the OAG was 52&amp;percnt; (51/98) vs. 45.5&amp;percnt; (30/66); P &amp;gt; 0.05. The clinical pregnancy rate per embryo transfer was 52.5&amp;percnt; in the OAG and 40&amp;percnt; in the NOA group (P &amp;gt; 0.05). It was concluded from the results of this study that both the BDTB and TESE were useful diagnostic and prognostic predictors of successful sperm retrieval for ICSI treatment in the non-obstructive and obstructive azoospermic male patients. The clinical embryo implantation rates were not significantly different between both groups (NOA group and OAG) which may indicate that the embryos of both groups have similar implantation potentials. Bilateral differential testicular biopsies were found to be positively correlated with the TESE and ICSI outcomes.

Anti Mullerian Hormone and Inhibin B: Discriminatory Markers of Azoospermia?

Anti mullerian hormone (AMH) and inhibin B are synthesised and secreted by the male gonads. The objective of this ongoing study is to investigate if circulating concentrations of AMH and/or inhibin B could be useful in discriminating obstructive and non-obstructive azoospermia in men. In a prospective study, blood samples were taken from men with non-obstructive azoospermia (n=10), obstructive azoospermia (n=8) and fertile controls (n=19). Blood samples were collected in plain tubes and serum was separated and frozen until assays were carried out. AMH and inhibin B were measured using commercial assays. AMH concentrations in the non-obstructive azoospermia males were ∼50% lower (mean ±stdev; 4±2.4ng/ml, P=0.003) than the controls (9.3±5.2ng/ml). Levels of AMH in the obstructive azoospermia group were not significantly altered (6.4±4.6ng/ml) from the control or the non-obstructive azoospermia males. Inhibin B levels were undetectable in 7 patients (<10pg/ml) and 25pg/ml in one patient in the non-obstructive azoospermia group. The levels of inhibin B in the obstructive azoospermia males (175±107pg/ml) were significantly higher (∼17 fold, P<0.001) than the non-obstructive azoospermia males. Serum inhibin B in the control group was 181±58.5pg/ml similar to the obstructive azoospermia males. Serum AMH and inhibin B are significantly decreased in non-obstructive azoospermia compared to the controls. However, this pilot study shows that inhibin B is better than AMH in discriminating between obstructive and non-obstructive azoospermia.

Comparison of the outcome of intracytoplasmic sperm injection in obstructive and non‐obstructive azoospermia in the first cycle: a report of case series and meta‐analysis

To investigate the outcome of intracytoplasmic sperm injection with fresh and cryopreserved-thawed testicular spermatozoa in the first cycle in patients with obstructive azoospermia (OA) and non-obstructive azoospermia (NOA), a total of 90 cases, 48 OA and 42 NOA were studied. All patients underwent sperm retrieval by testicular sperm extraction (TESE) while their wives received conventional ovarian hyperstimulation. The hormone levels, testicular histology, the rates of sperm retrieval, fertilization, implantation and pregnancy were analysed and evaluated. This study and other four similar studies were subjected to meta-analysis. Sperm retrieval was successful in 100% OA and 61% NOA. Fresh spermatozoa were used in 87.5% and 92.4% of OA and NOA cases respectively; while cryopreserved-thawed spermatozoa were used in 12.5% and 7.6% of OA and NOA, respectively. The fertilization, implantation and clinical pregnancy rates were 65.5%, 15% and 25% respectively in OA group, and 54.2%, 5% and 23.1% respectively in NOA group. Sperm status (fresh or thawed), male partner's age, female age and male serum follicle-stimulating hormone had no significant effect upon fertilization rate, implantation rate, or pregnancy rate per embryo transfer. The results of meta-analysis indicate that there is no statistically significant difference in clinical pregnancy rates between the two groups. There was a significantly higher fertilization rate among OA patients in all analysed studies (95% CI = 14.29-15.71, d.f. 832, T = 1.96). In conclusion, although the fertilization rate was significantly higher in the OA group in our study and from the given meta-analysis, there were some differences as regards pregnancy rates. Although the overall effect was more or less similar pregnancy rates in both subtypes of azoospermia, this may not be true if non-male infertility variables were controlled for in all studies.

Etiology of azoospermia in 100 consecutive nonvasectomized men

History was taken systematically for 100 azoospermic, nonvasectomized men referred consecutively to a Danish fertility clinic. The men were examined by ultrasound, and their blood samples were analyzed for karyotype, Y microdeletions, and cystic fibrosis transmembrane conductance regulator gene mutations. In 29% of patients, the condition could be explained by genetic abnormalities; in 22%, by diseases or external influence; and in 27%, by former cryptorchidism. The azoospermic condition remained unexplained in only 22%.

ANALYSIS OF LIPID PEROXIDATIVE LEVELS IN SEMINAL PLASMA OF INFERTILE MEN BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

For the purpose to evaluate the significance of lipid peroxidative products on male infertility, the levels of malondialdehyde (MDA), which is one of the final products of lipid peroxidation in seminal plasma, were determined. Ninety-three male infertile patients were divided into obstructive azoospermic group (12 cases), non-obstructive azoospermic group (15 cases), oligozoospermic group (21 cases), asthenozoospermic group (19 cases), oligoasthenozoospermic group (16 cases) and oligoasthenoteratozoospermic group (10 cases). Eighteen fertile males were included in the control group. MDA concentrations of seminal plasma in the fertile and infertile men were detected by high-performance liquid chromatography (HPLC). The results showed that the concentration of MDA in seminal plasma differed significantly between the control group and all the infertile groups (P < 0.01) except the obstructive azoospermic group, between the oligoasthenozoospermic group and the oligozoospermic and asthenozoospermic groups (P < 0.01), and between the oligoasthenoteratozoospermic group and the oligozoospermic and asthenozoospermic groups (P < 0.01). MDA concentration of seminal plasma in the oligoasthenoteratozoospermic group differed significantly from that in the oligoasthenozoospermic group (P <0.05). The results suggested that detection of MDA concentrations in seminal plasma by HPLC has an indicative value on the diagnosis of male infertility induced by overproduction of reactive oxygen species in male reproductive system.

Pregnancy outcome and neonatal data of children born after ICSI using testicular sperm in obstructive and non-obstructive azoospermia.

Registries on outcome of ICSI pregnancies obtained with testicular sperm do not differentiate between obstructive (OA) and non-obstructive azoospermia (NOA). We evaluated the pregnancy outcome and neonatal data on children born after ICSI using testicular sperm of men with histologically proven OA or NOA. Pregnancies obtained after ICSI using testicular sperm of men with defined NOA (n = 70) were compared with those of men with OA (n = 204). Multiple birth rates in NOA and OA couples, respectively, were 21 versus 27% (P = NS), overall preterm delivery rates were 38 versus 26% (NS), and prematurity rates were 24 versus 13% for singletons (NS) and 86 versus 54% for twins (relative risk 1.59, 95% confidence interval 1.04-2.42). Median gestational age for singletons was 38.3 versus 39.3 weeks, respectively (P < 0.05). The low birth weight rates were 34 versus 31%, respectively (NS). The early perinatal mortality rate was 66 versus 15 per 1000 births, respectively, (NS). Major congenital malformations were observed in 4 versus 3%, respectively, of the live born babies (NS). Prenatal karyotypes showed 7% de-novo abnormalities in the NOA group versus 1% in the OA group (NS). Our data do not show differences between NOA and OA pregnancies except for a strong tendency towards a lower gestational age in singletons and a higher percentage of premature twins in the NOA group. Although our data are based on a limited sample, the differences observed call for further analysis. Given the low pregnancy rates after ICSI with NOA, a multicentre study, differentiating NOA and OA patients, would be recommended.

Cleavage stage versus blastocyst transfer in ICSI cycles of azoospermic couples.

Cleavage stage versus blastocyst transfer in ICSI cycles of azoospermic couples.

The origin of spermatozoa does not affect intracytoplasmic sperm injection outcome

Objective: To assess whether the origin of spermatozoa, ejaculate or testicular, affects intracytoplasmic sperm injection (ICSI) outcome. Study design: Retrospective study of 890 consecutive first ICSI and embryo transfer cycles done for male infertility. The ICSI outcome of ejaculated spermatozoa ( n=780) and testicular spermatozoa retrieved from patients with obstructive azoospermia ( n=43), non-obstructive azoospermia ( n=53) and severe oligoasthenoteratozoospermia ( n=14) were compared by using chi-square test, independent t-test and ANOVA with Bonferroni test. Results: All azoospermic males had a diagnostic testicular biopsy at least 6 months before the ICSI procedure. Spermatozoa were successfully retrieved in all 43 patients with obstructive azoospermia and in 72.6% of 73 non-obstructive cases. The cycle characteristics of the four groups were similar apart from a younger mean female age in the non-obstructive azoospermia group when compared with the ejaculated spermatozoa group. The fertilization, implantation and clinical pregnancy rates were comparable among the four groups. Conclusion: Testicular spermatozoa recovered from patients with obstructive and all types of non-obstructive azoospermia were as much as effective as ejaculated spermatozoa in ICSI.

Proportion of cystic fibrosis gene mutations not detected by routine testing in men with obstructive azoospermia.

Infertile men with obstructive azoospermia may have mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, many of which are rare in classic cystic fibrosis and not evaluated in most routine mutation screening. To assess how often CFTR mutations or sequence alterations undetected by routine screening are detected with more extensive screening in obstructive azoospermia. Routine screening for the 31 most common CFTR mutations associated with the CF phenotype in white populations, testing for the 5-thymidine variant of the polythymidine tract of intron 8 (IVS8-5T) by allele-specific oligonucleotide hybridization, and screening of all exons through multiplex heteroduplex shift analysis followed by direct DNA sequencing. Male infertility clinic of a Canadian university-affiliated hospital. Of 198 men with obstructive (n = 149) or nonobstructive (n = 49; control group) azoospermia, 64 had congenital bilateral absence of the vas deferens (CBAVD), 10 had congenital unilateral absence of the vas deferens (CUAVD), and 75 had epididymal obstruction (56/75 were idiopathic). Frequency of mutations found by routine and nonroutine tests in men with obstructive vs nonobstructive azoospermia. Frequency of mutations and the IVS8-5T variant in the nonobstructive azoospermia group (controls) (2% and 5.1% allele frequency, respectively) did not differ significantly from that in the general population (2% and 5.2%, respectively). In the CBAVD group, 72 mutations were found by DNA sequencing and IVS8-5T testing (47 and 25, respectively; P<.001 and P = .002 vs controls) vs 39 by the routine panel (P<.001 vs controls). In the idiopathic epididymal obstruction group, 24 mutations were found by DNA sequencing and IVS8-5T testing (12 each; P=.01 and P=.14 vs controls) vs 5 by the routine panel (P=.33 vs controls). In the CUAVD group, 2 mutations were found by routine testing (P=.07 vs controls) vs 4 (2 each, respectively; P=.07 and P=.40 vs controls) by DNA sequencing and IVS8-5T testing. The routine panel did not identify 33 (46%) of 72, 2 (50%) of 4, and 19 (79%) of 24 detectable CFTR mutations and IVS8-5T in the CBAVD, CUAVD, and idiopathic epididymal obstruction groups, respectively. Routine testing for CFTR mutations may miss mild or rare gene alterations. The barrier to conception for men with obstructive infertility has been overcome by assisted reproductive technologies, thus raising the concern of iatrogenically transmitting pathogenic CFTR mutations to the progeny.