Mast cells in testicular biopsies of infertile men with 'mixed atrophy' of seminiferous tubules

Mast cells in the bilateral testicular biopsies of 30 patients with a 'mixed atrophy' of seminiferous tubules were analysed. Seven biopsies from vasectomized patients served as controls. With regard to their characteristic location within testicular tissue, two groups of mast cells could be distinguished, in both control and infertile patients: 'interstitial' mast cells (located between Leydig and other interstitial cells as well as in the vicinity of blood vessels) and 'peritubular' mast cells (located in the close proximity of the tubular lamina propria or incorporated in the lamina propria itself). Morphometric data indicated a significant increase in the number and volume of mast cells in infertile patients when compared with controls. In the biopsies of infertile patients that were analysed both 'interstitial' and 'peritubular' mast cells showed a significant increase in their number and volume, although it appeared that 'peritubular' mast cells increased at a higher rate than 'interstitial' mast cells. A significant negative correlation was found between the following variables: volume and number of mast cells, testis volume and the status of spermatogenesis evaluated by Johnsen's scoring. It was concluded that the increased presence of mast cells is closely associated with an impairment of spermatogenesis.     

Quantification of cyclin A1 and glyceraldehyde-3-phosphate dehydrogenase expression in testicular biopsies of infertile patients by fluorescence real-time RT-PCR

The objective of this study was to establish a classification of meiotic disorders by detection of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and cyclin A1 mRNA in patients presenting with non-obstructive azoospermia. GAPDH and cyclin A1 expression levels were detected in 37 histologically classified testicular tissue specimens by one-line RT-PCR. Levels of cyclin A1 expression (NCyclinA1) were high in tissue specimens with full spermatogenesis [3.519 +/- 3.141 (mean +/- SD)] but only minimal in those without haploid germ cells. A lack of cyclin A1 expression was seen in most sertoli-cell-only syndrome (SCOS) specimens. The criteria for an ideal internal standard were only partially met by GAPDH, as we detected decreased expression in tissue samples with maturation arrest [(268.2 +/- 106.2 relative gene expression (mean +/- SD)] and SCOS (201.7 +/- 95.2) compared to those with normal histological findings (325.1 +/- 129.3). It was concluded that the level of cyclin A1 mRNA expression predicts meiotic disorders during spermatogenesis. GAPDH is not an ideal internal standard for specific gene expression in testicular tissue specimens.     

Pathological observation on lamina propria of seminiferous tubules in infertile men

In this paper the authors observed the pathological alterationsin lamina propria of seminiferous tubules from 41 patients witholigospermia and azoospermia by using morphometry and immuno-histochemistry. Morphological alterations in seminiferous tubules ofthese patients were classified into three types; hypospermato-genesis, spermatogenic arrest and Sertoli-cell-only syndrome. Thelimiting membrane of the seminiferous tubules was found thicker invarious degrees. Morphometrically, there were significant differ-ences between the three types of alterations in terms of thickness ofthe lamina propria and the diameter of the seminiferous tubules.Immunohistochemically, the inner and outer layers of the laminapropria were positive for type Ⅳ collagen and laminin, while theintermediate layer, the hyperplastic zone, was negative, but thiszone was positive for type Ⅰ and Ⅲ collagen. The results indicatedthat the basic component of the altered lamina propria was of colla-gen fibers but not of type Ⅳ collagen, and     

不育症患者睾丸曲细精管界膜的病理学观察

应用形态计量学和免疫组织化学方法观察４１例少精子症和无精子症患者的睾丸活检曲细精管界膜的病理学变化。曲细精管生精上皮的病理变化主要有三种类型：生精功能低下、生精阻滞和唯支持细胞综合征。三种病变的曲细精管均表现有不同程度的界膜增厚。形态计量学结果显示三种病变的曲细精管界膜厚度、管内径与正常对照相比均有显著性差异，管内径与界膜厚度呈负相关。免疫组化层粘连蛋白Ⅳ型胶原染色显示界膜的内外层呈阳性反应，而中间增厚带为阴性，Ⅰ型、Ⅲ型胶原免疫组化染色界膜增厚的中间带为阳性。提示：病变界膜的成分为胶原纤维，但与正常成分－Ⅳ型胶原不同，表现为Ⅰ型及Ⅲ型胶原增生。     

Presence of IL-18 in testicular tissue of fertile and infertile men

Recently, IL-18 was identified in human testes. Moreover, an inverse correlation was found between the levels of IL-18 and the number and motility of spermatozoa. We examined the presence of IL-18 protein in normal and impaired spermatogenesis. Testicular tissue specimens were taken from 25 nonobstructive azoospermic patients undergoing testicular sperm extraction and from autopsies of three healthy controls. The presence of IL-18 in human testicular cells was examined by immunohistochemical staining of paraffin-embedded sections, using a specific antibody for human IL-18. In testicular tissue of healthy controls as well as in study cases, presence of IL-18 was identified in somatic, mitotic, meiotic and post-meiotic cells in correlation with their presence. In all patients, Leydig cells were less intensively stained. Mitotic cells were immunostained in the control group and less intensively in hypospermatogenesis and maturation arrest subgroups. Primary spermatocytes were in general most efficiently stained. The expression of IL-18 mRNA (as examined by real-time PCR analysis) showed significantly lower expression in testicular tissues with impaired spermatogenesis when compared to normal tissues. We report the first study demonstrating the presence of IL-18 in human testicular tissue at the protein level. The presence of this cytokine in somatic as well as in different types of germ cells may suggest its involvement in the regulation of the spermatogenic process and steroidogenesis under physiological and pathological conditions.     

Leptin expression in the testicular tissue of fertile and infertile men

There have been several studies about the presence of leptin in serum and testicular tissue, and none of them compares the leptin expression in the testicular tissue of fertile and infertile men. We assessed the presence of leptin expression in the testicular tissue of fertile and infertile men. 20 azoospermic infertile men were included in the study. All patients underwent testicular sperm extraction (TESE) for ICSI. For the detection of leptin, the immunohistochemistry was carried out. Intensity of immunohistochemical staining was subjectively estimated and expressed as negative (-), weak positive (+), intermediate positive (++) and strong positive (+++). Testicular tissues of 5 fertile patients, aged 50-60 years, was stained with leptin for control group. Mann-Whitney U test was used as the statistical method. There was no statistically significant difference in leptin staining between infertile patients and control group (p < 0.05). Leptin staining in tubuli seminiferi and Leydig cells were generally equal or Leydig cells were stained (+) much. This difference was not statistically significant. We found that there is leptin staining in Leydig cells and tubuli seminiferi. There is no difference in normal and infertile men for leptin staining properties in testicular tissue. This condition suggests that the effect of leptin on reproductive functions originates from a systemic effect related to central neuroendocrine system, androgen levels or spermatogenic existence rather than its direct effect on testicular tissue.     

Stage-specific degeneration of germ cells in the seminiferous tubules of non-obese diabetic mice

The cause of fertility problems in insulin-dependent diabetes is largely unknown. To evaluate the role of autoimmunity-associated phenomena in the testis as a possible cause of the derangement in spermatogenesis, the stage-specific apoptosis of germ cells in the insulitis phase of pre-diabetes was quantified in the testes of non-obese diabetic (NOD) mice. The seminiferous epithelium of normal BALB/c and NOD mice contained cells positive for in-situ end-labelling (ISEL) of DNA. ISEL-positive germ cells formed clusters in the seminiferous epithelium of the NOD mice in marked contrast to the seminiferous epithelium of the BALB/c mice, which contained only individual cells positive for ISEL. ISEL-positive cells were present in the basal and luminal compartments of the epithelium. Ultrastructural analysis and demonstration of externalized phosphatidyl serine confirmed that the cells were undergoing apoptosis. The ultrastructurally apoptotic cells included spermatogonia, spermatocytes and spermatids. In cytological squash preparations of segments of seminiferous tubules from NOD mice aged 17–20 weeks, the number of ISEL-positive cells/mm tubule was significantly lower in segments at stages I–II of the seminiferous epithelial wave but higher at stages III–IV in comparison to BALB/c mice. The numbers of ISEL-positive cells/mm tubule in the other stages were similar in the two strains of mice. Analysis of ³²P-3' -end labelled DNA from the testes showed that the BALB/c mice had relatively more DNA fragmentation than did the NOD mice. These data suggest that autoimmune insulitis in the NOD mice is associated with increased amounts and abnormal stage distribution of apoptosis in the seminiferous epithelium, resulting in derangement of spermatogenesis.     

Prevalence of carcinoma in situ in testicular biopsies of infertile Iranian men

Almost all testicular germ cell tumours are proved to originate from carcinoma in situ cells. Infertility is one of the factors that increase the risk of carcinoma in situ. The reported prevalence for carcinoma in situ from different parts of the world is 0–3.7% in infertile men. This retrospective study was performed to determine the prevalence of carcinoma in situ in Iranian infertile men. We reviewed the testicular biopsies of 1153 infertile men at the pathology department of Avicenna Infertility Center. One hundred and fifty‐one cases were suspicious of having carcinoma in situ. Immunohistochemical marker for placental alkaline phosphatase was employed to confirm the diagnosis of carcinoma in situ. Positive results were detected in 7 (0.6%) of 1153 cases (95% CI 0.24%–1.24%), 6 (0.94%) of which (95% CI 0.34%–2.04%) were under the age of 35 years (636 patients were in this age group). This study is the first study in Iran determining the prevalence of carcinoma in situ among the infertile Iranian men; the result is in the range of reports from other countries.     

Adenosine deaminase activity in fertile and infertile men

Adenosine deaminase (ADA; E.C.3.5.4.4) catalyses the deamination of adenosine to inosine. In the human reproductive system, the importance of enzymes that affect metabolism of adenosine, particularly adenosine deaminase, has been noticed. The purpose of this study was to determine the plasma activities of total adenosine deaminase (ADAT), and its isoenzymes, ADA1 and ADA2, in fertile and infertile men. Plasma activities of ADA and its isoenzymes were measured in 55 fertile men and 70 infertile men. There was a significant difference in the ADA1 and ADA2 activities between fertile and infertile individuals (P < 0.01). The activity of ADAT, ADA2 and ADA1 in infertile men was higher than that in fertile individuals. This alteration in ADA activity can lead to reduced adenosine levels, which may be involved in disturbing the fertility process.     

Ultrastructure of the seminiferous tubules in oligoasthenoteratozoospermic men associated with varicocele

Varicocele is associated with venous reflux that may cause increased heat and interstitial pressure within the testes, with variable pathological effects on spermatogenesis. This study aimed to study the ultrastructural testicular changes in the seminiferous tubules of 20 infertile severe oligoasthenoteratozoospermia (OAT) men associated with varicocele and five patients with obstructive azoospermia without varicocele as controls. They were subjected to testicular biopsy which was evaluated by transmission electron microscopy. Ultrastructurally, the seminiferous epithelium in the testicular biopsies of infertile severe OAT men associated with varicocele was variably affected in the form of thickening of the peritubular connective tissue, vacuolation of Sertoli cell and germ cell cytoplasm, presence of degenerated and apoptotic cells among the germinal epithelium, altered spermatids and abnormal spermatozoa. It is concluded that varicocele in severe OAT men is associated with ultrastructural changes in the seminiferous tubule.     

Expression of leptin and leptin receptor in the testis of fertile and infertile patients

The aim of our study was to investigate the relationships between the expression of leptin, leptin receptor in the testis and spermatogenesis, and testosterone (T) concentration in infertile men. Testicular tissue samples were collected from the testes of five fertile volunteers, eight patients with obstructive azoospermia (OA), six patients with Sertoli cell-only syndrome (SCO) and 32 oligospermic patients with varicocele testis. In testicular tissue, leptin and leptin receptor were identified by staining with polyclonal antibodies. Serum follicle stimulating hormone, lutenising hormone (LH), and T were determined by chemiluminescence assays. Leptin was expressed on germ cells, mainly on spermatocytes. The ratio of immunostained germ cells to total germ cells was inversely correlated with the concentration of T (r = -0.32, P = 0.01), sperm concentration (r = -0.51, P = 0.002) and Johnsen's score (r = -0.44,P = 0.005). In contrast, leptin receptor immunostained cells were found in the interstitium, primarily in Leydig cells. Leptin receptor expression on Leydig cells was inversely correlated with serum T concentration (r = -0.50, P < 0.001). The dysfunction of spermatogenesis is associated with an increase in leptin and leptin receptor expression in the testis.     

Quantification of survivin mRNA in testes of infertile patients and in testicular germ cell tumours: high levels of expression associated with normal spermatogenesis

Deregulated apoptosis of germ cells may contribute to male infertility as well as malignant transformation. Survivin, an inhibitor of apoptosis (IAP), is overexpressed in all the most common human malignancies, but barely detectable in normal tissues. We used real-time polymerase chain reaction (PCR) to quantify survivin mRNA expression in normal testes (n = 22), testes with defective spermatogenesis (n = 26) and testicular germ cell tumours (TGCTs; n = 16). Survivin was expressed at high levels in normal testes. Testicular survivin levels in infertile patients were related inversely to the severity of spermatogenic failure (p < 0.001), with a lack of expression in most specimens with pre-meiotic spermatogenic arrest and in all those with germ cell aplasia. Lower levels of expression were observed in TGCTs than in normal testes. While survivin expression was detected in most TGCTs with undifferentiated components (12 of 13), it was absent in all mature teratomas (n = 3). These data show that survivin is expressed in normal and transformed germ cells. Its downregulation in spermatogenic disorders indicates that survivin may contribute to the normal balance between germ cell proliferation and apoptosis. In TGCTs, survivin expression appears to be lost with somatic differentiation.     

Assessment of testicular volume: A comparison of fertile and sub-fertile West African men

BackgroundWhile the semen analysis appears to be the cornerstone in the evaluation of testicular function, the testicular volume has long been associated with testicular function. However, racial variations in testicular volume do exist. Neither the critical minimum testicular volume that guarantees adequate function, nor the optimal testicular volume that indicates peak testicular function are also known.ObjectiveTo evaluate the relationship between testicular volume and function using scrotal ultrasound scan in black West African men.Patients and methodsThe study examined 236 subjects over a period of one year. The subjects comprised of 136 patients with diagnosis of male infertility, as well as 100 healthy individuals as control. The relevant clinical history of each patient was extracted from their case notes. All the subjects had their testes examined using a high frequency (7.5 mHz) linear transducer of an ultrasound scanner. The results were expressed as percentages and tests of significance were done using the chi-square and Student's t-test. A P-value < 0.05 was considered statistically significant.ResultsThe mean testicular volume for the sub-fertile patients was 15.32 ml while it was 19.89 ml in the control group. There was a statistically significant difference between the testicular volumes in fertile and infertile men at different age groups, while there was an inverse relationship between testicular volume and severity of oligospermia. This was, however, not directly linear as a mean testicular volume of 18–20 ml was associated with highest semen density. Volumes higher than 20 ml and lower than 18 ml were associated with reduced sperm density. There was also a sharp decline in sperm density when the mean testicular volume reduced from 14 ml to 13 ml. Severe oligospermia (<5 million/ml) was associated with mean testicular volume less than 12 ml.ConclusionTesticular volume on scrotal ultrasound correlates well with severity of oligospermia in men with sub-fertility. While the critical mean testicular volume necessary for adequate spermatogenesis has not been determined, it appears there is an optimal testicular volume of 18–20 ml at which spermatogenesis is at its peak in sub-fertile Nigerian men.     

Seminal fibronectin-like antigen and transferrin concentrations in infertile and fertile men

Summary.  Fibronectin like antigen (Fn) and transferrin (Trs) levels were measured in the seminal plasma of 40 fertile and 102 infertile men. The concentrations of both proteins were significantly ( P <0.001) higher in the fertile controls compared to the infertile groups. The levels of Fn and Trs (mean value ± SEM) in the fertile men were 857.9 ± 9.8 μg ml^-1 and 164.0 ± 6.5 μg ml^-1, respectively; in the azoospermic men ( n = 17) 552.7 ± 24.65 μg ml^-1 and 20.7 ± 2.19 μg ml^-1, respectively; in the group of severe oligozoospermia ( n = 35) 568.34 ± 25.7 μg ml^-1 and 31.1 ± 4.18 μg ml^-1, respectively; in the moderate oligozoospermic group ( n = 8) 572.50 ± 47.9 μg ml^-1 and 43.4 ± 15.4 μg ml^-1 respectively, and in the asthenozoospermic group ( n = 26) 512.76 ± 40.4 μg ml^-1 and 47.0 ± 7.9 μg ml^-1, respectively. Of special interest was the finding from a group of 16 normospermic men (partners of couples with unexplained infertility) who showed significantly lower levels of Fn like antigen, 632.5 ± 26.9 μg ml^-1 ( P <0.001) and Trs 41.8 ± 6.94 μg ml^-1 ( P <0.0001) compared to normals. No correlation was found between Fn levels with either Trs or FSH levels or sperm count. In conclusion, our results indicate that male infertility is associated with changes in seminal plasma Fn like antigen concentrations and that it can be possibly used as an index of sperm fertilizing capacity.     

Mast cells in testicular biopsies of azoospermic men

This work aimed at identifying mast cells in testicular biopsies from 10 normal fertile controls, 20 patients with obstructive azoospermia and 70 patients with nonobstructive azoospermia. The biopsies were stained with haematoxylin and eosin stain for tubular-modified Johnson score and with toluidine blue stain for mast cells. Two populations of mast cells, peritubular and interstitial, were demonstrated in all sections with varied counts. Testicular sections with Sertoli cell only and spermatogenic arrest patterns demonstrated a significant increase in both peritubular and interstitial mast cells compared with other groups, whereas obstructive azoospermia demonstrated a nonsignificant increase compared with the controls. Mast cell count was significantly correlated negatively with Johnson score for both peritubular (P = 0.001) and interstitial (P = 0.001) populations. Whether these results could be a cause or an effect, a special role might be assigned to mast cells in the pathogenesis of disturbed spermatogenesis.     

Oxidative stress status and sperm DNA fragmentation in fertile and infertile men

Evidence suggests that disturbing the balance between reactive oxygen species levels and antioxidant contents in seminal plasma leads to oxidative stress resulting in male infertility. This study was carried out to identifying clinical significance of seminal oxidative stress and sperm DNA fragmentation in treatment strategies of male infertility in southwest Iran. Sperm parameters, lipid peroxidation and activity of antioxidant enzymes were assessed in fertile (n = 105) and infertile (n = 112) men. Malondialdehyde (MDA) levels in seminal plasma were found to be higher significantly (p < .001) in patients. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities in seminal plasma were significantly (p < .001) lower in infertile men. Significant negative correlations were observed between MDA levels and sperm motility and normal morphology. Spermatozoa with fragmented DNA were higher (p < .001) in infertile men and significantly correlated with MDA levels and SOD and GPx activities. MDA of 4.2 nmol/ml, SOD of 4.89 U/ml and GPx of 329.6 mU/ml were optimum cut‐off limits to discriminate infertile patients from fertile men. The results show the leading role of oxidative stress in aetiology of male infertility in southwest Iran and indicate that evaluation of seminal antioxidant status and DNA integrity can be helpful in men attending infertility clinics during fertility assessment.     

Assessment of seminal plasma laminin in fertile and infertile men

Aim： To assess laminin levels in the seminal plasma of infertile and fertile men, and to analyze the correlation of laminin levels with sperm count, age, sperm motility and semen volume. Methods： One hundred and twenty-five recruited men were equally divided into five groups according to their sperm concentration and clinical examination： fertile normozoospermia, oligoasthenozoospermia, non-obstructive azoospermia （NOA）, obstructive azoospermia （OA） and congenital bilateral absent vas deferens （CBAVD）. The patients＇ medical history was investigated and patients underwent clinical examination, conventional semen analysis and estimation of seminal plasma laminin by radioimmunoassay. Results： Seminal plasma laminin levels of successive groups were： 2.82 ± 0.62, 2.49 ± 0.44, 1.77 ± 0.56, 1.72 ± 0.76, 1.35 ± 0.63 U/mL, respectively. The fertile normozoospermic group showed the highest concentration compared to all infertile groups with significant differences compared to azoospermic groups （P 〈 0.05）. Testicular contribution was estimated to be approximately one-third of the seminal laminin. Seminal plasma laminin demonstrated significant correlation with sperm concentration （r = 0.460, P 〈 0.001） and nonsignificant correlation with age （r = 0.021, P = 0.940）, sperm motility percentage （r = 0.142, P = 0.615） and semen volume （r = 0.035, P = 0.087）. Conelusion： Seminal plasma laminin is derived mostly from prostatic and testicular portions and minimally from the seminal vesicle and vas deferens. Estimating seminal laminin alone is not conclusive in diagnosing different cases of male infertility.     

Seminal leucocyte subpopulations and sperm function in fertile and infertile Chinese men

The level of seminal leucocytes and the prevalence of leucocytospermia was determined in a group of fertile and infertile southern Chinese men in Hong Kong. Sixteen normal fertile semen donors and 49 men with male factor infertility were studied prospectively. None had antisperm antibodies and past or present evidence of genital tract infection. Seminal leucocytes and their subsets were analysed using monoclonal antibodies and an immunocytochemical alkaline phosphatase-anti-alkaline phosphatase conjugate technique. Seminal leucocytes were detectable in 94% and 86% of the fertile and infertile men respectively, with the predominant subset being granulocytes. Leucocytospermia (> 1 × 10⁶ leucocytes/ml) was found in only one of the 49 (2%) infertile men without clinical evidence of genito-urinary infection. Inverse correlations were observed between (1) the percentage of spermatozoa with normal morphology and the number of T-helper/inducer cells, (2) the linearity of sperm movement and the number of T-lymphocytes. In conclusion, the level of seminal leucocytes and the prevalence of leucocytospermia is low in infertile Chinese subjects. The effect of seminal leucocytes on sperm function in these subjects needs further evaluation.     

Distribution of carcinoma-in-situ in testes from infertile men

Quantitative histologial studies on 4 testes removed because of carcinoma-in-situ (CIS) were performed in order to determine the distribution of CIS within the testis and to estimate the likelihood of diagnosing testicular CIS by biopsy. The CIS changes were distributed in all parts of the testes but were less frequent in the parts adjacent to the epididymis. In the 4 testes examined 1.4 to 599 of the entire testicular volume contained seminiferous tubules with CIS. In parts of the testes where more than approximately 10% of the testicular volume consisted of tubules with CIS all simulated biopsies measuring 3 mm contained the lesion. The same was true for simulated biopsies measuring 1.5 mm when more then approximately 30% of the testicular volume consisted of tubules with CIS.
If the distribution of CIS generally is similar to that found in the 4 analysed testes there seems to be a high probability of detecting the disease by one or two testicular biopsies.