Assessment of seminal YKL-40 in infertile men with varicocele

Several studies attempted to explain the negative impact of varicocele on spermatogenesis and fertilisation processes. YKL-40 is a novel glycoprotein biomarker that had been associated with several diseases. This quasi-interventional study aimed to assess the seminal levels of YKL-40 in infertile men with varicocele before and after varicocelectomy. Overall, 50 men were included in this study divided into 20 healthy fertile men and 30 infertile oligoasthenoteratozoospermic (OAT) men with varicocele that underwent varicocelectomy. All participants were subjected to history taking, clinical examination and scrotal Doppler. Also, semen analysis and seminal YKL-40 assessment were carried out in the start and 6 months after varicocele surgical repair. The results showed a significant increase in the mean seminal YKL-40 level in infertile OAT men with varicocele compared with the healthy fertile men. Six months post-varicocelectomy, the mean seminal KYL-40 level exhibited significant decreases correlated with improved sperm parameters. Overall, seminal levels of YKL-40 showed significant negative correlations with sperm concentration, total sperm motility and sperm normal morphology. It could be concluded that seminal YKL-40 is elevated in infertile OAT men with varicocele where varicocelectomy induces decreased seminal YKL-40 levels correlated with improved semen parameters.     

Seminal plasma reactive oxygen species-antioxidants relationship with varicocele grade

This work aimed to assess seminal plasma reactive oxygen species (ROS)-antioxidants relationship with varicocele grade in infertile men with oligoasthenoteratozoospermia (OAT). The study included 89 infertile OAT men with varicocele divided into grade I (n = 22) and grade II (n = 43), grade III (n = 24) and compared with 20 healthy fertile controls. In their seminal plasma, two ROS parameters (malondialdehyde [MDA], hydrogen peroxide [H(2) O(2) ]) and four antioxidants (superoxide dismutase [SOD], catalase [Cat], glutathione peroxidase [GPx], vit.C) were estimated. There was significant increase in seminal MDA, H(2) O(2) and significant decrease in seminal SOD, Cat, GPx, vit.C in varicocele-associated OAT cases when compared with the controls. Compared with grade I cases, varicocele cases with grades II, III demonstrated significant increase in estimated seminal MDA, H(2) O(2) and significant decrease in seminal SOD, Cat, GPx, vit.C. It is concluded that seminal oxidative stress (OS) is related to increased varicocele grade in infertile OAT men associated with varicocele.     

不育男性双侧Ⅰ度精索静脉曲张对睾丸体积和生殖激素水平的影响

目的 评价不育男性双侧I度精索静脉曲张对睾丸体积和生殖激素水平的影响.方法 185例不育男性双侧I度精索静脉曲张(A组)和149例正常生育男性(B组),比较其睾丸体积、卵泡刺激素(FSH)、黄体生成素(LH)和睾酮(T)水平.结果 A组患者两侧睾丸体积均小于B组,但睾丸体积绝对差异和睾丸体积相对差异与B组比较,无统计学意义.A组患者血清FSH水平高于B组,而LH、T与B组相比,差异无统计学意义.结论 不育男性双侧I度精索静脉曲张可导致患者双侧睾丸体积减小,血清FSH水平升高.     

Assessment of seminal granulysin in infertile men with varicocele

Varicocele has a common association with male infertility, but its exact role is still debated. Apoptosis has been suggested as one of the mechanisms of varicocele‐associated infertility. Granulysin is a molecule that plays a role in apoptosis with no previous study about its role in male infertility. This case‐controlled study aimed to assess seminal plasma granulysin level in infertile patients with varicocele. This study involved 90 men that were allocated into fertile normozoospermic men (n = 20), infertile men without varicocele (n = 30) and infertile men with varicocele (n = 40). These men were subjected to history taking, clinical examination, semen analysis and estimation of seminal granulysin. In general, seminal granulysin level was significantly elevated in infertile men compared with fertile men. Infertile men with varicocele showed significantly higher seminal granulysin compared with infertile men without varicocele, in bilateral varicocele cases and in grade III varicocele. Seminal granulysin level was negatively correlated with sperm concentration, sperm motility, sperm normal forms percentage and testicular volumes. It is concluded that increased seminal granulysin has a negative impact on spermatogenesis in infertile men in general and in infertile men associated with varicocele in particular.     

Chromosomal abnormalities and y chromosome microdeletions in infertile men with varicocele and idiopathic infertility of South Indian origin

Various factors cause spermatogenesis arrest in men and, in a large number of cases, the underlying reason still remains unknown. Little attention is paid to determining the genetic defects of varicocele-related infertility. The objective of our present study was to investigate the chromosomal abnormalities and Y chromosome microdeletions in infertile men of South Indian origin with varicocele and idiopathic infertility. Metaphase chromosomes of 251 infertile men with varicocele and unexplained infertility were analyzed using Giemsa-Trypsin-Giemsa (GTG) banding and fluorescence in situ hybridization (FISH). The microdeletions in 6 genes and 18 sequence-tagged-sites (STS) in the Yq region were screened using polymerase chain reaction (PCR) techniques. Out of 251 infertile men, 57 (22.7%) men were with varicocele, of which 8.77% were azoospermic, 26.31% were severely oligozoospermic, 21.05% were mildly oligozoospermic, and 43.85% were oligoasthenoteratozoospermic (OAT), and 194 (77.29%), with idiopathic infertility, of which 51% were azoospermic, 13.40% were severely oligozoospermic, 19.07% were mildly oligozoospermic, and 16.4% were with OAT. Genetic defects were observed in 38 (15.13%) infertile individuals, including 14 (24.56%) men with varicocele and 24 (12.37%) men with idiopathic infertility. The frequencies of chromosomal defects in varicocele and idiopathic infertility were 19.3% and 8.76%, respectively, whereas Y chromosome microdeletions were 5.26% and 3.60%, respectively. Overall rate of incidence of chromosomal anomalies and microdeletions in 251 infertile men were 11.5% and 3.98%, respectively, indicating a very significant higher association of genetic defects with varicocele than idiopathic male infertility. Our data also demonstrate that, among infertile men with varicocele, severely oligozoospermic and OAT men with varicocele have higher incidences of genetic defects than mildly oligozoospermic and azoospermic men.     

Programmed cell death in varicocele-bearing testes

Accelerated apoptosis is a significant factor in the pathophysiology of male infertility disorders associated with abnormal spermatogenesis. This study aimed to investigate apoptosis in varicocele-bearing testes. Sixty four men with varicocele (18 fertile and 46 infertile) were studied compared with eight men with obstructive azoospermic as controls. Apoptosis was assessed in testicular biopsy specimens using terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) method as well as electron microscopy. The results demonstrated that the occurrence of apoptotic changes comprised all types of germ cells but not affecting Sertoli cells. Mean tubular apoptotic indices of fertile or infertile men with varicocele were significantly higher than controls (mean ± SD 4.55 ± 1.03%, 6.29 ± 1.82% versus 2.71 ± 0.45%, P  < 0.05). Mean Leydig cells apoptotic indices of infertile men with varicocele were significantly higher than those of fertile men without varicocele as well as controls (1.18 ± 0.38%, 0.68 ± 0.15%, 0.31 ± 0.21%, P  < 0.05). Apoptotic indices were nonsignificantly correlated with Johnsen score, testicular volume or varicocele grade. It is concluded that testicular apoptosis is increased in varicocele-associated men either fertile or infertile who may be implicated in associated spermatogenic dysfunction.     

Seminal cyclooxygenase relationship with oxidative stress in infertile oligoasthenoteratozoospermic men with varicocele

This study aimed to assess the relation of seminal cyclooxygenase COX‐1, COX‐2 with oxidative stress in infertile oligoasthenoteratozoospermic (OAT) men with varicocele (Vx). In all, 128 men were allocated into fertile men, fertile men with Vx, infertile OAT men without Vx and infertile OAT men with Vx. They were subjected to history taking, clinical examination and semen analysis. Also, seminal COX‐1, COX‐2, malondialdehyde (MDA) and glutathione peroxidase (GPx) were estimated. Mean levels of seminal COX‐1, COX‐2 were over‐expressed, the mean level of seminal MDA was significantly increased, and the mean level of seminal GPx was significantly decreased in infertile OAT men with Vx compared with other groups. Seminal COX‐1 and COX‐2 were over‐expressed in cases with Vx grade III compared with Vx grades I, II cases and in cases with bilateral Vx compared with unilateral Vx. There was significant negative correlation between seminal COX‐1 and COX‐2 with sperm concentration, sperm motility, sperm normal morphology, seminal GPx and significant positive correlation with seminal MDA. It is concluded that seminal COX‐1 and COX‐2 are over‐expressed in infertile OAT men with Vx compared with fertile men with/without and infertile OAT men without Vx being associated with oxidative stress, Vx grade and Vx laterality.     

Expression of endothelial nitric oxide synthase in the Sertoli cells of men with infertility of various causes

OBJECTIVE: To investigate how endothelial nitric oxide (eNOS) expression in the seminiferous tubules might be related to spermatogenesis, by examining eNOS expression in testicular tissue of patients infertile from various causes. PATIENTS AND METHODS: The study included five fertile men with a normal sperm concentration, nine patients with obstructive azoospermia, 20 with varicocele testes and eight with idiopathic azoospermia (Sertoli cell-only syndrome). Testicular biopsy specimens were examined by immunohistochemistry for eNOS protein expression, in addition to a routine pathological assessment. eNOS protein was detected using an eNOS monoclonal antibody. A Sertoli cell staining index (SSI) was defined as the ratio of stained Sertoli cells per total number of Sertoli cells, and was compared among the groups. RESULTS: eNOS was localized to Sertoli cells in the seminiferous tubules and Leydig cells in the interstium; although some degenerating germ cells stained, normal germ cells did not. The SSI was significantly lower in patients with Sertoli cell-only syndrome than in either fertile men or patients with obstructive azoospermia or varicocele. However, the SSI did not correlate significantly with the Johnsen score. CONCLUSION: The expression of eNOS in Sertoli cells may depend on the existence of germ cells and be associated with germ cell development.     

Seminal SIRT1-oxidative stress relationship in infertile oligoasthenoteratozoospermic men with varicocele after its surgical repair

This work assessed seminal SIRT1-oxidative stress (OS) relationship in infertile oligoasthenoteratozoospermic (OAT) men after varicocele repair. Overall, thirty OAT men with varicocele were investigated. Inclusion criteria were infertile males (males who were unable to initiate a pregnancy within 1 year of regular unprotected intercourse), confirmed OAT and normal female factor. These cases were subjected to history taking, clinical checkup and semen analysis. In their semen, seminal SIRT1, malondialdehyde (MDA) and glutathione peroxidase (GPx) levels were assessed. These men were subjected to varicocele surgical repair and were followed up for 3 months. Post-operatively, the mean seminal SIRT1, GPx levels showed significant increases and the mean MDA level showed significant decrease compared to the pre-operative levels linked to improved sperm parameters. The mean seminal SIRT1, GPx, MDA levels showed more significant improvement in grade III varicocele cases compared to grade II cases after surgical repair. Seminal SIRT1 levels showed significant positive correlations with sperm concentration, sperm motility, sperm normal morphology, seminal GPx levels and a significant negative correlation with seminal MDA levels. It could be concluded that seminal SIRT1 is significantly decreased in infertile OAT men with varicocele after its surgical repair linked to improved sperm parameters as well as seminal OS.     

A morphometric and ultrastructural study of the changes in the lamina propria in adolescents with varicocele.

OBJECTIVE: To verify if changes in the lamina propria and its components, reported in adults with varicocele, are already present in adolescents with idiopathic varicocele, using a morphometric and ultrastructural study. MATERIALS AND METHODS: Twenty testicular biopsies from adolescents (mean age 15.6 years) were obtained during surgery for left idiopathic varicocele; five testicular biopsies from adolescents (mean age 16 years) undergoing surgery for hydrocele or inguinal hernia were used as controls. Biopsy specimens were processed for light and transmission electron microscopy; the sections were evaluated morphometrically using computerized image analysis. RESULTS: Morphometric and ultrastructural examination of the testes with varicocele showed an increased thickness in the lamina propria, caused principally by an increase in the extracellular matrix components, and deep invaginations towards the germinal epithelium. CONCLUSION: There is detectable damage of the lamina propria in adolescents affected by left idiopathic varicocele, although not as well developed or as severe as in adults.     

Apoptosis-related proteins in the testes of infertile men with varicocele

OBJECTIVES: To assess immunohistochemically the expression of apoptosis-related proteins in the testes of infertile men, to determine which of these proteins were related to hypospermatogenesis, as a previous report suggested that apoptosis was suppressed in infertile men with varicocele. MATERIALS AND METHODS: The expression of Bcl-2, Bax, caspase-1 (ICE) and caspase-3 (CPP32) were examined in bilateral testicular specimens from 26 infertile men with varicocele and six normal testicular specimens, using the avidin-biotin-peroxidase complex method. Clinical variables were also assessed. RESULTS: Bax, ICE, and CPP32 were expressed in germ cells, while Bcl-2 was not. Differences in staining in left or right testes were not significant. In both testes of infertile patients with varicocele, significantly fewer germ cells stained for CPP32 than in controls (P < 0.001). For Bax and ICE, total germ cell staining was similar between these groups. Staining was less frequent in infertile patients for both CPP32 and ICE when the analysis was restricted to spermatogonia. Serum luteinizing hormone levels correlated positively with CPP32 staining (P = 0.0457). CONCLUSIONS: The reduced expression of CPP32 participates in regulating apoptosis in the testes of infertile men with varicocele.     

Semen and reproductive hormone parameters in fertile men with and without varicocele

Although varicoceles are a widely accepted identifiable male factor in infertile couples, the benefit of varicocele repair in improving pregnancy and live birth rates remains uncertain. The Study for Future Families obtained semen and reproductive hormone samples from US men whose partners were currently pregnant. In our analysis cohort of 709 men, a varicocele was detected by clinical examination in 56 (8%) of men. Men with varicocele had smaller left testis, and lower total and total motile sperm counts than men without varicocele. Gonadotropin levels were higher as well in men with varicocele. Interestingly, testosterone levels were also slightly higher in men with varicocele. Despite these differences, there was no difference between the groups in the time to achieve the study pregnancy or percentage of men with a previous pregnancy. We conclude that even in fertile men, varicoceles are associated with some degree of testicular hypofunction. This would support current recommendations to consider varicocele repair in male partners in infertile couples who demonstrate both a varicocele and abnormal semen parameters and after evaluation for treatable female factors.     

Hormone profile in infertile men

Plasma basal levels of FSH, LH, prolactin, and testosterone were evaluated in 263 men with fertility problems. Significant increases in FSH were detected in all groups of infertile men except those with infection, varicocele, infection and varicocele, and obstructive azoospermia. No differences in LH were detected in all groups, except those with chromosomal testicular abnormality and idiopathic seminiferous tubular failure where significant increases were revealed. No differences in prolactin were detected, while a decrease was found in testosterone in the group with chromosomal testicular abnormality. FSH seems to be a most valuable hormone index in the routine workup of male infertility, while prolactin does not offer substantial information. The combined LH and testosterone evaluation might be of diagnostic importance in selected cases.     

精索静脉曲张大鼠生精小管生精上皮的超微结构研究

目的 通过精索静脉曲张大鼠睾丸的透射电镜观察,了解生精上皮更多的细胞病理现象.方法 雄性Sprayue-Dawley大鼠30只,随机分为(1)精索静脉曲张组(VG)20只,(2)假手术对照组(SOG)10只,按Saypol方法建立左精索静脉曲张模型,处死观察大鼠,取其左、右侧睾丸组织,作透射电镜观察.结果 VG组双侧睾丸均出现生精小管生精上皮的结构损害,左侧相对较重.与SoG组比较,VG的生精上皮变化表现为:支持细胞胞质内出现大量空泡,大量溶酶体出现(72%vs 28%);紧密连接破坏(69%vs31%),线粒体数量减少(160 vs 362), 两者均有显著性差异(P＜0.01);精子顶体形成异常(62%vs38%),尾部线粒体鞘部分缺失,嵴肿胀(71%vs 29%),尾部横切面线粒体数目减少(186 vs 401), 两者均有显著性差异(P＜0.01).结论 生精小管生精上皮的结构损害是由精索静脉曲张导致男性不育的重要原因.     

Decrease of topoisomerase I activity in the testes of infertile men with varicocele

Topoisomerase I, which is involved in DNA metabolism, was measured in the testes of 37 infertile men with varicocele, using a minute amount of testicular biopsy specimen. In 38% of infertile men topoisomerase I activity was lower than that of normal men. Forty-six percent of patients with high grade varicocele (grade III) showed reduced topoisomerase I activity, while only 26% of patients with low grade varicocele (grades I and II) showed reduced levels of topoisomerase I. Johnsen's score of the patients with low topoisomerase I activity was lower than that of patients with normal levels of topoisomerase I. The decrease of topoisomerase I activity may be correlated with at least some fraction of hypospermatogenesis associated with varicocele.     

The role of reactive oxygen species in testicular dysfunction associated with varicocele

OBJECTIVE: To determine the level of malondialdehyde (MDA), an indirect indicator of lipid peroxidation-induced injury by reactive oxygen species, in testicular biopsy specimens from infertile patients with and without varicocele. PATIENTS AND METHODS: Levels of MDA were measured in the testicular biopsy specimens from 25 infertile men (15 with varicocele, mean age 30.0 years, SD 5.7, range 23-45, and 10 without, mean age 28.7 years, SD 4.2, range 21-34). All patients were evaluated by a detailed history, physical examination, semen analysis (at least twice), serum follicle-stimulating hormone and free testosterone levels, testicular biopsy and contact imprint. Scrotal colour Doppler ultrasonography was used to confirm suspected varicocele. The level of MDA in testicular biopsy specimens was measured using the thiobarbituric acid test and the results expressed per unit tissue weight. RESULTS: As a causal factor for infertility, varicocele was identified in 15 men (60%), testicular failure in four (16%), idiopathic infertility in four (16%) and obstruction in two (8%). Of the 15 patients with varicocele, eight had bilateral varicocele and it was subclinical in three; the varicocele was grade I in four, grade II in six and grade III in two. The mean (SD) MDA level in the men with a subclinical varicocele was 15.7 (3.1) pmol/mg tissue, while in those with grade I-III varicocele it was 32.9 (12.25), 37.1 (12.25) and 86.9 (2.89) pmol/mg tissue, respectively. The levels in patients with grade III varicocele were significantly greater than in the other groups (P < 0.05). The mean MDA level in patients with or without varicocele was 38.3 (22.92) and 33.5 (18.93) pmol/mg tissue, respectively (P > 0.05). CONCLUSION: These results suggest that increasing levels of MDA are associated with higher grades of varicocele and support a possible rationale for controlled trials in infertile men with varicocele.     

Potential role of reactive oxygen species on testicular pathology associated with infertility

Aim: To investigate the level of malondialdehyde (MDA), a direct indicator of lipid peroxidation-induced injury by reactive oxygen species (ROS), in testicular biopsy specimens from infertile patients. Methods: Levels of MDA were measured in testicular biopsy specimens from 29 consequent-randomized infertile men, aged 29.58±4.76 (21-45) years. All patients were evaluated by a complete medical and reproductive history, physical examination, semen analysis (at least two), serum follicle-stimulating hormone and free testosterone levels, testicular biopsy and contact imprint. Scrotal colour Doppler ultrasonography was used to confirm suspected varicocele. The testicular MDA level was measured using the thiobarbituric acid test and the results were expressed per unit tissue weight. Results: As a causal factor in infertility, varicocele was identified in 17 (58.6 %) patients, and idiopathic infertility, testicular failure and obstruction in 4 (13.8 %) patients each. The testicular MDA level was 13.56 (6.01),     

Radiologically assessed testicular changes in infertile males with varicocele

The aim of this study was to assess vascular disturbances in the testis of infertile males associated with varicocele. In total, 124 consecutive male subjects were divided into the following groups: healthy fertile controls (n = 10), Gp1 (n = 28); infertile males with subclinical varicocele, Gp2 (n = 26); infertile males with grade I left varicocele, Gp3 (n = 28); infertile males with grade II left varicocele and Gp4 (n = 32); infertile males with grade III left varicocele. They were subjected to colour duplex for pampiniform plexus and scrotal scintigraphy. There was significant decrease in arterial blood velocity, testicular arterial diameters and testicular perfusion especially in high grade varicocele compared with healthy controls. The mean vein diameter demonstrated significant negative correlation with arterial diameter, arterial blood velocity, perfusion index, testicular size and significant positive correlation with perfusion index. The mean testicular size demonstrated significant positive correlation with arterial blood velocity, perfusion index and nonsignificant correlation with arterial diameter. It is concluded that there are significant decreases in testicular volume, testicular perfusion, blood velocity and testicular artery diameter in infertile males with varicocele.     

Elevation of intratesticular and scrotal skin surface temperature in men with varicocele

A possible mechanism of varicocele-induced infertility is believed to be elevation of testicular temperature. Sensitive needle thermistors were used to measure directly intratesticular and bilateral scrotal surface temperatures simultaneously in anesthetized infertile men with varicocele and control subjects. We found that intratesticular temperature is elevated significantly in humans with varicocele. In addition, we have shown that scrotal skin surface temperature is elevated in men with varicocele. Furthermore, we demonstrate that unilateral varicocele is associated with bilateral elevation of scrotal surface temperature. These findings confirm the results of animal studies revealing elevation of intratesticular temperature associated with varicocele and suggest bilateral elevation in unilateral varicocele.     

Increased testicular 8-hydroxy-2'-deoxyguanosine in patients with varicocele

OBJECTIVES: To assess immunohistochemically 8-hydroxy-2'-deoxyguanosine (8-OHdG, a sensitive biological marker of oxidative damage and stress) in testicular tissue from patients with varicocele, to determine whether 8-OHdG is related to spermatogenesis, as oxidative stress in testicular DNA is associated with poor semen quality and reduced fertility. PATIENTS, SUBJECTS AND METHODS: Testicular tissue samples were collected from the testes of 36 patients with varicocele and five normal volunteers. The mean (sd) age of the patients was 35.3 (5.4) years. The prevalence of immunostained germ cells among all the germ cells in >20 seminiferous tubules was calculated. RESULTS: The mean (sd) sperm concentration and Johnsen's score in the varicocele group was lower than in the normal group, at 27.6 (11.8) vs 80.4 (21.7) million/mL (P < 0.001) and 7.6 (1.1) vs 9.0 (0.7), respectively (P < 0.05). In all patients, 8-OHdG was mainly expressed in the spermatogonia and spermatocytes in the seminiferous tubule. Immunostained germ cells correlated negatively with sperm concentration (r = -0.48, P = 0.01) and Johnsen's score (r = -0.46, P = 0.02), but not with follicle-stimulating hormone, luteinizing hormone or testosterone level. Immunostained germ cells were significantly more prevalent in the varicocele group, at 53.6 (5.7)%, than that in the normal group, at 14.3 (2.3)% (P < 0.001). The prevalence of immunostained germ cells and clinical grades were positively related; the ratio recorded for varicocele grades I, II and III was 38.0 (10.0)%, 41.0 (9.1)%, and 57.0 (9.3)%, respectively (grade I vs grade III, P < 0.05). CONCLUSIONS: Increased 8-OHdG expression in the testis was associated with deficient spermatogenesis in infertile men with varicocele.