Obstructive Azoospermia Group Research Articles

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.

Comparison of ART outcome between non-obstructive azoospermia and obstructive azoospermia: 15 years experiences

Comparison of ART outcome between non-obstructive azoospermia and obstructive azoospermia: 15 years experiences

Human zygotes respond to sperm DNA damage by slowing DNA replication

OBJECTIVE: Some authors describe similar and high fertilization and pregnancy rates in non-obstructive azoospermia (NOA) patients whereas others report lower outcomes in NOA than in obstructive azoospermia (OA) patients. Sperm DNA damage is associated with reduced fertilization rates, embryo quality and pregnancy rates and higher rates of spontaneous miscarriage and childhood disease. Meanwhile, mouse zygotes respond to spermDNA damage through a non-apoptoticmechanism that acts by slowing paternal DNA replication and embryonic development. In this study, we compared the early development of embryos fertilized with NOA and OA spermatozoa by using time-lapse monitoring system. DESIGN: This is a retrospective study. MATERIALS AND METHODS: Six oocytes with ICSI using testicular spermatozoa from NOA and 35 oocytes with OA from 2013 to 2014 were used. The exact times for each embryo division and developmental parameters were calculated in hours after ICSI. Time-lapse images of each embryo were retrospectively analyzed by PrimoVision software. The developmental events observed in each embryo were at the time of the visibility of two pronuclei (2PN), syngamy, two cells and four cells. These parameters were compared between the NOA group and the OA group. RESULTS: The time of the visibility of 2PN in the NOA group (9.5 1.4 hours) was significantly earlier than that in the OA group (12.1 2.3 hours). However, the time from the visibility of 2PN to the syngamy in the NOA group (13.2 3.4 hours) was significantly longer than that in the OA group (10.6 2.7 hours). The time between the syngamy and the two cells in the NOA group (12.2 1.3 hours) was comparable with that in the OA group (16.6 10.2 hours). The time from the two cells to the four cells in the NOA group (13.5 3.5 hours) was also comparable with that in the OA group (10.0 4.2 hours). CONCLUSION: The time from the visibility of 2PN to the pronuclear fading (syngamy) after ICSI with testicular spermatozoa from NOAwas prolonged compared to that after ICSI with testicular spermatozoa from OA.

Relationships between sperm DNA fragmentation, age of donors and patients and children with psychic disorders

OBJECTIVE: Some authors describe similar and high fertilization and pregnancy rates in non-obstructive azoospermia (NOA) patients whereas others report lower outcomes in NOA than in obstructive azoospermia (OA) patients. Sperm DNA damage is associated with reduced fertilization rates, embryo quality and pregnancy rates and higher rates of spontaneous miscarriage and childhood disease. Meanwhile, mouse zygotes respond to spermDNA damage through a non-apoptoticmechanism that acts by slowing paternal DNA replication and embryonic development. In this study, we compared the early development of embryos fertilized with NOA and OA spermatozoa by using time-lapse monitoring system. DESIGN: This is a retrospective study. MATERIALS AND METHODS: Six oocytes with ICSI using testicular spermatozoa from NOA and 35 oocytes with OA from 2013 to 2014 were used. The exact times for each embryo division and developmental parameters were calculated in hours after ICSI. Time-lapse images of each embryo were retrospectively analyzed by PrimoVision software. The developmental events observed in each embryo were at the time of the visibility of two pronuclei (2PN), syngamy, two cells and four cells. These parameters were compared between the NOA group and the OA group. RESULTS: The time of the visibility of 2PN in the NOA group (9.5 1.4 hours) was significantly earlier than that in the OA group (12.1 2.3 hours). However, the time from the visibility of 2PN to the syngamy in the NOA group (13.2 3.4 hours) was significantly longer than that in the OA group (10.6 2.7 hours). The time between the syngamy and the two cells in the NOA group (12.2 1.3 hours) was comparable with that in the OA group (16.6 10.2 hours). The time from the two cells to the four cells in the NOA group (13.5 3.5 hours) was also comparable with that in the OA group (10.0 4.2 hours). CONCLUSION: The time from the visibility of 2PN to the pronuclear fading (syngamy) after ICSI with testicular spermatozoa from NOAwas prolonged compared to that after ICSI with testicular spermatozoa from OA.

A genome-wide DNA methylation study in azoospermia

The objective of this study was to assess genome-wide DNA methylation in testicular tissue from azoospermic patients. A total of 94 azoospermic patients were recruited and classified into three groups: 29 patients presented obstructive azoospermia (OA), 26 displayed non-obstructive azoospermia (NOA) and successful retrieval of spermatozoa by testicular sperm extraction (TESE+) and 39 displayed NOA and failure to retrieve spermatozoa by TESE (TESE-). An Illumina Infinium Human Methylation27 BeadChip DNA methylation array was used to establish a testicular DNA methylation pattern for each type of azoospermic patient. The OA and NOA groups were compared in terms of the relative M-value (the log2 ratio between methylated and non-methylated probe intensities) for each CpG site. We observed significantly different DNA methylation profiles for the NOA and OA groups, with differences at over 9000 of the 27578 CpG sites; 212 CpG sites had a relative M-value >3. The results highlighted 14 testis-specific genes. Patient clustering with respect to these 212 CpG sites corresponded closely to the clinical classification. The DNA methylation patterns showed that in the NOA group, 78 of the 212 CpG sites were hypomethylated and 134 were hypermethylated (relative to the OA group). On the basis of these DNA methylation profiles, azoospermic patients could be classified as OA or NOA by considering the 212 CpG sites with the greatest methylation differences. Furthermore, we identified genes that may provide insight into the mechanism of idiopathic NOA.

The impact of surgical sperm retrieval on the in vitro fertilization outcomes of infertile patients with temporary ejaculation failure on their oocyte retrieval day

To evaluate the impact of testicular sperm aspiration on the clinical outcomes of IVF patients with temporary ejaculation failure (TEF) on the day of oocyte retrieval. A case-control study. University-affiliated IVF center. One hundred eight infertile couples encountering TEF on the day of oocyte retrieval. Testicular sperm aspiration and intracytoplasmic sperm injection (TESA-ICSI) were employed to help the TEF couples to complete their IVF treatments. A group of obstructive azoospermia (OA) patients was chosen as a control group to evaluate the efficiency and safety of this intervention. Fertilization rates, transferable embryo rates, and clinical pregnancy rates (PRs). Through the TESA-ICSI, TEF couples achieved a 38.83% (40/103) clinical PR per IVF cycle. Compared with the OA group, the TEF group showed a comparable fertilization rate (66.43% vs. 65.25%), transferable embryo rate (60.76% vs. 60.85%), and PR (51.61% vs. 50.00%) in thawed ET cycles but a relatively low PR (25.49% vs. 44.64%) in fresh ET cycles. TESA-ICSI is a relatively efficient and safe way to help the patients encountering TEF.

Initial exploration of epididymal sperm nuclear maturity in obstructive azoospermia patients

To ascertain the epididymal sperm nuclear maturity in obstructive azoospermia (OA) patients. A total of 81 infertile males from andriatry clinic of Renji hospital were selected, including 32 OA patients (OA group) and 49 asthenospermia patients (asthenospermia group). Another 32 fertile males were recruited for the control group. All semen samples of the asthenospermia and control groups underwent computer aided semen analysis (CASA). For the OA group, sperm specimens were collected by percutaneous epididymal sperm aspiration (PESA) and semen analyses completed by manual method. Further acridine orange test and aniline blue stain assay were performed. The results were processed with SPSS 15.0. No statistically significant differences existed in semen volume and sperm density between the asthenospermia and control groups (both P > 0.05). The OA group was significantly lower than the control group in sperm density (P < 0.01). The asthenospermia and OA groups were also significantly lower than the control group in sperm motility (both P < 0.01). As to the percentage of grade a + b sperm, the asthenospermia group was also significantly lower than the control group (P < 0.01). Compared with the control group (81% ± 9%, 85% ± 8%), the negative rate of acridine orange test and aniline blue assay of OA (57% ± 20%, 64% ± 20%) and asthenospermia group (62% ± 19%, 67% ± 15%) were statistically lower (all P < 0.05). The sperm nuclear maturity is lower in the asthenospermia and OA groups than that in the control group. Male fertility should be further improved.

Analysis of the outcome of intracytoplasmic sperm injection using fresh or frozen sperm

• To compare the outcome of first-attempt intracytoplasmic sperm injection (ICSI) ICSI--embryo transfer (ET) cycles using frozen-thawed testicular sperm (FTTS), fresh testicular sperm (FTS), frozen-thawed epididymal sperm (FTES) and fresh epididymal sperm (FES) so as to determine which of these has the most successful ICSI outcome with respect to fertilization rate (FR), pregnancy rate (PR) and birth rate. • To assess the outcomes according to the underlying aetiology of azoospermia. • The records of 493 patients undergoing first-attempt ICSI between 1993 and 2008 were reviewed retrospectively. FTS was used in 112 cycles, FTTS in 43 cycles, FES in 279 cycles, and FTES in 59 cycles. • Within each group, the aetiology of the azoospermia was recorded according to history, clinical examination and histological analysis (n = 316). • The FR, clinical PR and delivery rate were calculated for each group with respect to the type of sperm retrieval used. • Analysis of the data showed no significant differences between any of the four groups in the FR, PR or delivery rate (P > 0.05). • There were no significant differences seen between fresh sperm (FTS and FES) and frozen sperm (FTTS and FTES) or between epididymal sperm (FES and FTES) and testicular sperm (FTS and FTTS) in any of the outcomes measured (P > 0.05). However, subset analysis showed a statistically higher FR and PR for FTTS over fresh sperm. • When comparing aetiologies, there was no significant difference in the FR, clinical PR and delivery rate between obstructive azoospermia (OA) and non-obstructive azoospermia (NOA) groups. However, sub-set analysis showed a higher PR and birth rate for FTTS over fresh sperm in both OA and NOA groups. • The results of the present study suggest that using frozen sperm in ICSI cycles is a reliable and favourable method with the same outcome as fresh sperm. • Testicular and epididymal sperm have similar ICSI outcomes for both fresh and frozen samples. However, results suggest a tendency for higher PRs and birth rates for frozen than for fresh testicular sperm in both OA and NOA aetiologies. • The aetiology of azoospermia does not significantly affect the outcome of first-attempt ICSI. The higher rates in the frozen groups suggest that these patients have had better quality semen when they were initially harvested and frozen.

Assessment of Seminal Estradiol and Testosterone Levels as Predictors of Human Spermatogenesis

The proposed hypothesis for this study was that seminal testosterone/estradiol levels and/or their ratios may be a good indicator for predicting normal spermatogenesis. The concentrations of estradiol and testosterone in seminal fluid were measured using competitive immunoassay techniques in specimens collected from 192 infertile patients and 103 normospermic men. Infertile patients were subdivided into three groups according to their semen analysis results and testicular biopsy: oligozoospermia, obstructive azoospermia (OA), and nonobstructive azoospermia (NOA). Results showed that seminal testosterone levels in the infertile groups were lower than in the normospermic individuals (P < .01), whereas seminal estradiol levels in the OA group were significantly higher than those in normospermic and NOA groups (P < .01). Testosterone/estradiol ratios in the seminal plasma from the infertile groups were significantly lower than that in the normospermic group (P < .01). However, seminal estradiol levels among normospermic and NOA groups showed no significant differences. These results suggest that the local balance between androgen and estrogen, or their ratios, may play an important role in maintaining normal spermatogenesis. Also, decreased seminal testosterone/estradiol ratio may be a good indicator for identifying the absence of sperm production in NOA patients.

Assisted reproductive technology outcomes in azoospermic men: 10 years of experience with surgical sperm retrieval

An azoospermic man suffers from an absence of sperm in the ejaculate and this condition is present in about 10% of infertile men. Obstructive azoospermia (OA) is characterized by an occlusion or partial absence of the reproductive tract with the presence of normal spermatogenesis. On the other hand, non-obstructive azoospermia (NOA) is characterized by impaired spermatogenesis. In these cases, spermatozoa can be obtained by percutaneous epididymal or testicular sperm aspiration (PESA and TESA, respectively) and used for intracytoplasmic injection (ICSI). To compare ICSI outcomes using spermatozoa that were surgically retrieved by PESA and TESA, azoospermic patients were divided into the following categories: (i) TESA-NOA (n = 102), (ii) TESA-OA (n = 103), and (iii) PESA-OA (n = 171). Fertilization, pregnancy, and implantation rates were compared between the groups. We noted a lower normal fertilization rate (p = 0.0017) and a higher abortion rate (p = 0.0387) among men in the TESA group who had OA when compared with men in the PESA group who had OA. On the other hand, a lower normal fertilization rate (p = 0.05) and a lower rate of non-cleaved embryos (p = 0.034) was found in the TESA group of NOA patients as compared to the TESA group of OA patients. No statistically significant differences were detected between the TESA and PESA groups and the OA and NOA groups, respectively. The clinical outcomes of embryos arising from ICSI cycles using spermatozoa harvested via PESA and TESA were similar, regardless of whether the patient had obstructive or non-obstructive azoospermia.

Genome-wide expression of azoospermia testes demonstrates a specific profile and implicates ART3 in genetic susceptibility.

Infertility affects about one in six couples attempting pregnancy, with the man responsible in approximately half of the cases. Because the pathophysiology underlying azoospermia is not elucidated, most male infertility is diagnosed as idiopathic. Genome-wide gene expression analyses with microarray on testis specimens from 47 non-obstructive azoospermia (NOA) and 11 obstructive azoospermia (OA) patients were performed, and 2,611 transcripts that preferentially included genes relevant to gametogenesis and reproduction according to Gene Ontology classification were found to be differentially expressed. Using a set of 945 of the 2,611 transcripts without missing data, NOA was further categorized into three classes using the non-negative matrix factorization method. Two of the three subclasses were different from the OA group in Johnsen's score, FSH level, and/or LH level, while there were no significant differences between the other subclass and the OA group. In addition, the 52 genes showing high statistical difference between NOA subclasses (p < 0.01 with Tukey's post hoc test) were subjected to allelic association analyses to identify genetic susceptibilities. After two rounds of screening, SNPs of the ADP-ribosyltransferase 3 gene (ART3) were associated with NOA with highest significance with ART3-SNP25 (rs6836703; p = 0.0025) in 442 NOA patients and 475 fertile men. Haplotypes with five SNPs were constructed, and the most common haplotype was found to be under-represented in patients (NOA 26.6% versus control 35.3%, p = 0.000073). Individuals having the most common haplotype showed an elevated level of testosterone, suggesting a protective effect of the haplotype on spermatogenesis. Thus, genome-wide gene expression analyses were used to identify genes involved in the pathogenesis of NOA, and ART3 was subsequently identified as a susceptibility gene for NOA. These findings clarify the molecular pathophysiology of NOA and suggest a novel therapeutic target in the treatment of NOA.

Sperm defect severity rather than sperm source is associated with lower fertilization rates after intracytoplasmic sperm injection

To evaluate the impact of sperm defect severity and the type of azoospermia on the outcomes of intracytoplasmic sperm injection (ICSI). This study included 313 ICSI cycles that were divided into two major groups according to the source of spermatozoa used for ICSI: 1) Ejaculated (group 1; n = 220) and 2) Testicular/Epididymal (group 2; n = 93). Group 1 was subdivided into four subgroups according to the results of the semen analysis: 1) single defect (oligo-[O] or astheno-[A] or teratozoospermia-[T], n = 41), 2) double defect (a combination of two single defects, n = 45), 3) triple defect (OAT, n = 48), and 4) control (no sperm defects; n = 86). Group 2 was subdivided according to the type of azoospermia: 1) obstructive (OA: n = 39) and 2) non-obstructive (NOA: n = 54). Fertilization (2PN), cleavage, embryo quality, clinical pregnancy and miscarriage rates were statistically compared using one-way ANOVA and Chi-square analyses. Significantly lower fertilization rates were obtained when either ejaculated sperm with triple defect or testicular sperm from NOA patients (63.4 +/- 25.9% and 52.2 +/- 29.3%, respectively) were used for ICSI as compared to other groups ( approximately 73%; P < 0.05). Epididymal and testicular spermatozoa from OA patients fertilized as well as normal or mild/moderate deficient ejaculated sperm. Cleavage, embryo quality, pregnancy and miscarriage rates did not differ statistically between ejaculated and obstructive azoospermia groups. However, fertilization, cleavage and pregnancy rates were significantly lower for NOA patients. Lower fertilization rates are achieved when ICSI is performed with sperm from men with oligoasthenoteratozoospermic and non-obstructive azoospermic, and embryo development and pregnancy rates are significantly lower when testicular spermatozoa from NOA men are used.

Application of percutaneous epididymal sperm aspiration and percutaneous testicle sperm aspiration in the differentiating diagnosis of obstructive and nonobstructive azoospermia

目的 探讨经皮附睾精子抽吸术(PESA)及经皮睾丸精子抽吸术(PTSA)在无精子症鉴别诊断中的应用价值.方法 采用PESA或PTSA对梗阻性无精子症(OA)组37例及非梗阻性无精子症(NOA)组28例进行附睾或睾丸取精,细胞学涂片进行分析.结果 OA组32例在附睾获得精子,5例在睾丸获得精子;NOA组7例在附睾获得精子,1例在睾丸获得精子.两组间获得的精子数差异显著(P＜0.01).结论 PESA和PTSA技术可用于OA和NOA的鉴别诊断。

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.

Sperm defect severity rather than sperm source is associated with lower fertilization rates after intracytoplasmic sperm injection

To evaluate the impact of sperm deficiencies and sperm source on the outcome of ICSI. Retrospective analysis of laboratory and clinical data. This study included 313 ICSI cycles performed from January 2002 to November 2003. Cycles were divided into two major groups according to the source of spermatozoa: Ejaculated (group I; n=220) and Testicular/Epididymal sperm (group II; n=93). Group I was subdivided into four groups according to the severity in the abnormality of the semen analysis: single defect (oligo-[O] or astheno-[A] or teratozoospermia[T], n=41), double defect (a combination of two single defects, n=45), triple defect (OAT, n=48), and control (no sperm defects; n=86). Group II was subdivided according to the cause of azoospermia: obstructive (OA: n=39) and non-obstructive (NOA: n=54). Sperm selection and microinjection were performed by a single operator using 400X magnification. Female age, mean number of oocytes retrieved, mean number of MII oocytes and mean number of embryos transferred were not statistically different among groups. Fertilization (2PN), cleavage, good embryo quality, clinical pregnancy and abortion rates were statistically compared using one-way ANOVA and Chi Square analyses, with P<0.05 considered significant. Significantly lower fertilization rates were obtained when either ejaculated sperm with triple defect or testicular sperm from NOA patients were used for ICSI as compared to other groups (Table 1, P<0.05). Epididymal and testicular spermatozoa from OA patients fertilized as well as normal or mild/moderate deficient ejaculated sperm. Cleavage, good embryo quality, pregnancy and abortion rates did not differ statistically between ejaculated and obstructive azoospermia groups. However, fertilization, cleavage and pregnancy rates were significantly lower for NOA patients. ICSI is a formidable therapy that can help most cases of male infertility. However, it bypasses pathological obstacles of the spermatozoa as a messenger rather than problems concerning the spermatozoa as a carrier. Lower fertilization potential is to be expected when ICSI is performed with spermatozoa from men with severe deficiencies, such as those with triple sperm defect and NOA. Such individuals may have increased genetic risks relative to other male factor conditions and their spermatozoa harbor defective messages that will not contribute to the development of a normal zygote.

Morphometrical quantification of spermatogonial germ cells with the 57B anti-MAGE-A4 antibody in the evaluation of testicular biopsies for azoospermia.

The melanoma-associated antigen (MAGE) gene family of cancer-testis antigens is expressed in certain malignant neoplasms and the testis, but not in other healthy tissues. The aim of this study was to determine the usefulness of immunohistochemical staining with the 57B anti-MAGE-A4 mouse monoclonal antibody (MAb) in testicular biopsy specimens from patients with nonobstructive azoospermia and obstructive azoospermia (OA). Fifty-four cases of Sertoli cell only (SCO), 30 cases of spermatocytic arrest, 15 cases of hypospermatogenesis, and 10 testicular biopsy specimens with OA (normal spermatogenesis) were evaluated. Immunohistochemistry was performed using the 57B MAb, which primarily recognizes the MAGE-A4 antigen in paraffinized tissues. The cells were quantitated by a computerized image analysis system. Testicular biopsy specimens with normal spermatogenesis exhibited strong nuclear and cytoplasmic MAGE-A4 staining of spermatogonia and weak staining of spermatocytes, but not spermatids or Sertoli or Leydig cells. No staining was detected in SCO cases. In five cases of SCO with focal spermatogenesis, spermatogonial cells that were initially missed by hematoxylin and eosin staining were detected by MAGE-A4 immunohistochemistry. Immunostaining with the 57B MAb greatly enhanced identification of spermatogonia in cases of spermatocytic arrest and hypospermatogenesis. The number of MAGE-A4-positive spermatogonia was significantly decreased in hypospermatogenesis, as opposed to the OA group (12.1 +/- 4.3 and 30.3 +/- 10.0, respectively). The number of MAGE-A4-positive primary spermatocytes was significantly increased in early maturation arrest, as compared with the OA group (48.2 +/- 10.8 and 16.9 +/- 9.8, respectively). The 57B anti-MAGE-A4 MAb is a useful marker for the detection and quantitation of spermatogonial germ cells. It also facilitates automated image analysis and provides greater accuracy in the histopathologic evaluation of testicular biopsy specimens.

Serum activin A and follistatin in disorders of spermatogenesis in men.

Inhibin, activin and follistatin are glycoprotein hormones produced by the gonads. Recent studies have shown that inhibin B is the predominant form of inhibin in the circulation in men. The objective of this study was to investigate circulating levels of activin A and follistatin in disorders of spermatogenesis in men and their relationship with FSH and inhibin B. Serum from five different groups of men was prospectively collected and stored at -20 degrees C. The groups were men with: (i) proven fertility (controls) (n=20), (ii) primary testicular failure (n=15), (iii) obstructive azoospermia (n=10), (iv) oligospermia (n=10) and (v) miscellaneous sperm dysfunction (n=40). WHO criteria (1992) were used for semen characterisation. Serum concentrations of 'total' activin A, follistatin, FSH and inhibin B were measured using specific two-site enzyme immunoassays. Activin A levels were significantly lower than in the controls in the obstructive azoospermia group and higher in the miscellaneous sperm dysfunction group. Serum follistatin levels did not significantly vary in any group compared with the controls. Circulating levels of FSH were higher than in the controls in the primary testicular failure and obstructive azoospermic group. Levels of inhibin B were lower than in the controls in all disorders of spermatogenesis studied. This study demonstrates that activin A and follistatin are in the circulation in males and activin A levels are significantly lower in obstructive azoospermia and higher in miscellaneous sperm dysfunction than in controls. The mechanism involved in altering the levels of activin A in these conditions is not clear. However, high follistatin:activin A molar ratios (>2.5) in all groups suggests that all activin A in the circulation is bound to follistatin in males.

Mutations of the cystic fibrosis gene and congenital absence of the vas deferens.

It is estimated that about 30 to 40% of couples seeking fertility treatments are diagnosed with male factor infertility. These males have a range of gonadal dysfunctions which include azoospermia (i.e., no sperm in the ejaculate); oligozoospermia (i.e. sperm count less than 20 million/ml), asthenozoospermia (i.e.sperm motility less than 50%) and teratozoospermia (i.e. sperm with normal morphology less than 30%). The group of patients with azoospermia represent about 25% of the total and, of these, about 30% have an obstructive process (obstructive azoospermia) while the remaining have a primary testicular failure (non-obstructive azoospermia). In the obstructive azoospermia group, 25% of males have congenital bilateral absence of the vas deferens (CBAVD), while the incidence among all infertile males is about 2%. In the USA, it is estimated that approximately 16000 males are affected by CBAVD. Anatomically, CBAVD is a disorder characterized by regression bilaterally of variable portions of the epididymis, vas deferens, and, in about 80% of cases, absence of the seminal vesicles. In about 10 to 20% of the patients, a renal anomaly is also present. These anatomical hallmarks are so strikingly similar to those observed in men with cystic fibrosis (CF) that, as early as 1971 (Holsclaw et al. 1971), these two apparently unrelated disorders were hypothesized to have the same genetic origin. The hypothesis was proven when mutations in the Cystic Fibrosis Transmembrane conductance Regulator (CFTR) gene (Kerem et al. 1989; Riordan et al. 1989;Rommens et al. 1989) were found in patients with CF as well patients with isolated CBAVD (Dumur et al. 1990; Anguiano et al. 1992; Patrizio et al. 1993).KeywordsCystic FibrosisCystic Fibrosis Transmembrane Conductance RegulatorCystic Fibrosis Transmembrane RegulatorCongenital AbsenceCystic Fibrosis Transmembrane Conductance Regulator MutationThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

The CFTR gene: male infertility and assisted conception

Approximately 30–40% of couples seeking fertility treatments have male factor infertility. Their dysfunctions include azoospermia, oligozoospermia, asthenozoospermia and teratozoospermia. Those with azoospermia represent about 25% of the total, and of these about 30% have an obstructive process while the remaining have either primary or secondary testicular failure. In the obstructive azoospermia group, 25% of males have congenital bilateral absence of the vas deferens (CBAVD).