Investigation of serum vitamin D and ischaemia-modified albumin levels in infertile Turkish men

Our aim was to explore the existence of a possible relationship of sperm motility with serum 25-hydroxyvitamin D3 (25-OH VD) levels and with ischaemia-modified albumin ( IMA) levels in infertile Turkish men. A total of 30 men with nonobstructive azoospermia (no spermatozoa in ejaculate), 30 men with oligospermia (total progressive motile sperm count (TPMSC) <15 × 10⁶/ml) and 33 fertile men with normospermia (with at least one child, as the control group) were enrolled in the study. The mean 25-OH VD levels for groups 1, 2 and 3 were 9.31 ± 6.46, 19.71 ± 12.80 and 30.52 ± 12.49 respectively (p < .05). There was a statistically significant difference in serum IMA levels among the groups (479.32 ± 307.56 vs. 296.37 ± 127.27 vs. 150.04 ± 81.05, respectively; p < .05). A positive correlation between serum 25-OH VD levels and TPMSC, and a negative correlation between TPMSC and serum IMA levels were determined. Infertile men had lower serum 25-OH VD and higher IMA levels than fertile men, with a positive correlation between serum 25-OH VD levels and TPMSC, and a negative correlation between TPMSC and serum IMA levels. Vitamin D supplementation may increase the sperm motility.     

Prediction of reproductive function recovery after microsurgical varicocelectomy in men from infertile couples: Clinical and laboratory predictors

The objective was to reveal predictors for fertility recovery after varicocelectomy in subfertile men. This retrospective study recruited 93 men with clinical varicocele and pathozoospermia who underwent microsurgical varicocelectomy. Stepwise discriminant analysis was performed to identify predictors of spontaneous pregnancy (SP) after surgery. ‘Clinically significant improvement’ (CSI) following varicocelectomy was defined as an increase in total progressively motile sperm count (TPMSC) by at least 12.5 million (calculated from WHO-2010 reference values). 52% of patients showed CSI, and 28% reported SP after surgery. Patients who reported SP (group II), compared to that remained infertile (group I), were younger (27.3 ± 2.9 versus 30.2 ± 4.2 years; p < .01), had less infertility period (24.1 ± 14.0 versus 44.4 ± 32.9 months; p < .05) and had initially higher TPMSC (median (25% −75%) = 34 (11–67) versus 9.5 (0–33) mln; p < .05). The stepwise discriminant analysis showed that male age (coefficient value = −0.157), total sperm motility (0.024) and postoperative increase in TPMSC (0.010) were the significant predictors of SP. The predictive ability, sensitivity and specificity of the discriminant function were 84%, 87%, 76% respectively. This algorithm can be recommended after varicocelectomy in decision-making on natural conception or the ART protocols usage.     

Glycerophosphocholine in seminal plasma of fertile and infertile men

Glycerophosphocholine (GPC) was measured in seminal plasma from 65 fertile men, 276 infertile men and 10 men before and after vasectomy, using a new enzymatic method. Extra-epididymal excretion of GPC accounted for 30% of the total seminal levels of GPC. From a diagnostic point of view, GPC determination did not appear to be a specific tool which could discriminate between secretory and excretory azoospermia. Although the seminal content of GPC was related positively to the total sperm count in both fertile and infertile men, there was an inverse relationship between the level of GPC and sperm motility when considering classes displaying the same total sperm count. This was observed in all classes from infertile men as well as in fertile men with a total sperm count lower than 200 x 10(6) sperm/ejaculate. These results suggest a possible role of GPC in the regulation of human sperm motility, which warrants further investigation.     

Seminal fibronectin in fertile and infertile males

This study aimed to assess seminal plasma fibronectin in fertile and infertile males. Ninety infertile males were investigated; asthenozoospermia (n = 27), asthenoteratozoospermia (n = 30), oligoasthenoteratozoospermia (n = 33) compared with 20 healthy fertile controls. They were subjected to semen analysis, seminal plasma fibronectin estimation by radial immune diffusion, serum testosterone (T) and follicle stimulating hormone (FSH) estimation by ELISA. There was significant increase of seminal plasma fibronectin among different infertile groups compared with the controls. Significant negative correlation was elicited between seminal fibronectin and sperm count, sperm motility grades A, B, A + B, sperm velocity, linear velocity, linearity index, sperm normal forms and serum T. Seminal fibronectin showed significant positive correlation with grade D sperm motility and serum FSH. ROC curve analysis discriminating controls and other infertile groups demonstrated criteria value of < 674 mg l(-1) (sensitivity 100% and specificity 96.4%). It is concluded that increased seminal fibronectin is associated with decreased sperm count and sperm motility.     

男性不育患者精子形态学分析与生殖激素关系的研究

目的:研究男性不育患者精子形态与生殖激素的关系,探讨畸形精子症的发病机制。方法:研究对象为90例男性不育患者,年龄25～40岁,利用Prader睾丸计评估患者睾丸容积,根据世界卫生标准进行精液常规分析,利用化学发光法测定血清生殖激素和性激素结合球蛋白(SHBG)浓度,计算得出游离睾酮和生物活性睾酮的浓度。结果:90例男性不育患者精子浓度均在正常范围,根据精子形态分析结果分为3个研究组,即组1(正常形态精子<4%)、组2(正常形态精子≥4%且<10%)和组3(正常形态精子≥10%),每组30例。3组之间年龄没有统计学差异(P>0.05);左侧睾丸容积分别为(14.27±3.65)ml,(16.90±3.57)ml和(14.57±3.57)ml,P组1,2=0.006,P组1,3=0.741和P组2,3=0.014;右侧睾丸容积分别为(14.60±3.70)ml,(16.60±3.35)ml和(14.67±3.54)ml,P=0.05;血清泌乳素(PRL)、卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E2)、总睾酮(TT)和SH-BG在3组之间均没有统计学差异(P>0.05);血清游离睾酮(FT)水平分别为(0.25±0.07)nmol/L,(0.29±0.07)nmol/L和(0.31±0.13)nmol/L,P组1,2=0.086,P组1,3=0.010和P组2,3=0.364;生物活性睾酮(Bio-T)水平分别为(5.81±1.58)nmol/L,(6.78±1.55)nmol/L和(7.29±3.02)nmol/L,P组1,2=0.086,P组1,3=0.010和P组2,3=0.364。另外,正常形态精子百分率与血清FT、Bio-T水平之间均存在正相关(P<0.05)。结论:男性不育患者血清FT和Bio-T水平越高,则正常形态精子百分率越高,提示FT和Bio-T可能参与畸形精子症发病过程。     

MicroRNA profiling in spermatozoa of men with unexplained asthenozoospermia

Asthenozoospermia (AZS) which is characterised by decreased sperm motility is one of the main causes of male infertility. Recent studies demonstrated altered microRNAs (miRNAs) in total semen, seminal plasma and spermatozoa of asthenozoospermic men. In line with these studies, it was aimed to evaluate the miRNA expression profile in spermatozoa of unexplained asthenozoospermic men. Thirty‐nine cases with idiopathic AZS and 35 fertile and healthy men as control were included. After total RNA extraction from spermatozoa, high‐throughput sequencing technology was employed to display miRNA profiles in spermatozoa samples pooled from AZS cases and healthy controls. Relative quantification by real‐time PCR was performed to validate RNA‐seq results. SNORD48 was used as normaliser gene, and fold change was calculated by 2^?ΔΔCt method. Profiling results showed that 18 altered miRNAs in AZS men in comparison to controls. Subsequently, seven miRNAs were selected to validate by RT‐PCR that showed MiR‐888‐3p significantly overexpressed in AZS cases (p = 0.014) in comparison with controls. It seems upregulation of miR‐888‐3p was associated with idiopathic AZS. This finding paves the way to the future investigation on the actual molecular role of miR‐888‐3p in aetiology of AZS.     

Age-related disease pattern in infertile men: increasing incidence of infections in older patients

In a cross-sectional retrospective study, the age-dependency of semen parameters of men with specific andrological diseases was investigated in 3,698 men (mean age 33 years; range 19-63 years) attending a tertiary referral clinic for couple infertility. In all, 1,853 patients suffered from idiopathic infertility (mean age 33.2 years; range 19-60 years), 388 from infections of the accessory glands (34.2; 20-61), 363 had a history of maldescended testes (32.0; 20-51) and 833 a varicocele (33.6; 20-63). In 172 patients, a varicocele and an infection (34.3; 24-54) and in 94 a varicocele and a history of maldecended testes (32.5; 21-53) were found. Semen analysis was performed according to WHO criteria. An age-related decrease in testicular volume was not observed in any group. Total sperm count decreased significantly with advancing age only in patients with an infection of the accessory glands. We found no evidence for age-dependent progressive deterioration of semen variables in the other groups. It may be concluded that the causes for the spermatogenetic defects in men with idiopathic infertility, a history of maldescended testes or varicocele are acquired before adolescence and do not progress during adulthood. In contrast, infections of the accessory glands may lead to decreased sperm counts by functional impairment or partial occlusion of the seminal ducts. Thus, if in the general population declining sperm counts are observed, genital tract infections may be suspected as a contributing factor.     

A controlled trial of the use of erythromycin for men with asthenospermia

A double-blind crossover controlled trial of Erythromycin was conducted in 78 men with asthenospermia (mean pretreatment sperm concentration greater than or equal to 20 X 10(6)/ml and motility less than 60%). Sperm motility increased above the pretreatment mean after the first one month course of treatment for both Erythromycin (7.9 +/- 2.6% P less than 0.01; mean +/- SEM) and the placebo group (7.5 +/- 2.2% P less than 0.01) and also after the second one month course of treatment, given one month after the first, for both Erythromycin (11.8 +/- 2.7% P less than 0.001) and placebo (11.4 +/- 2.8% P less than 0.001). There was no significant difference between the changes in sperm motility with Erythromycin and placebo. Twelve pregnancies occurred during the four month trial time--eight with Erythromycin and four with placebo (chi 2 = 2.51 P = NS). It is concluded that in infertile men, sperm motility tends to increase spontaneously with time. Erythromycin treatment has no significant effect on semen quality and a larger trial will be required to determine if Erythromycin increases fertility.     

中国乙型肝炎和非肝炎男性间的精液质量和辅助生殖技术治疗结果的比较

本研究旨在确定乙型肝炎病毒（HBV）感染对精子质量和辅助生殖技术治疗结局的影响。分析2008年1月至2009年12月于浙江大学附属妇产科医院寻求辅助生育治疗的916例男性（457例乙型肝炎病毒携带者和459例阴性对照男性）的精液参数;将2004年1月至2009年12月期间的男性乙肝病毒携带的587个常规体外受精（In vitro fertilisation,IVF）周期和325个单精子卵细胞浆内注射（Intracytoplasmic sperm injection,ICSI）周期作为阳性观察组,根据取卵日期、女方年龄及ART受精方式通过统计软件以1：1比例匹配,设立夫妇双方均为HBsAg阴性的对照组（587个IVF周期,324个ICSI周期）,分析两组间的辅助生殖治疗结局。与对照组相比,乙肝携带者表现有较低的精液体积、精子总数以及较差的精子活动力和形态学表现（P〈0．05）;乙肝男性夫妇接受ICSI治疗后,双原核（2PN）受精率、优质胚胎率、胚胎着床率以及临床妊娠率均显著性降低俨〈0．05）;而两组病人间的IVF治疗结局差异无显著性（P〉0．05）。通过Logistic回归分析发现乙肝病毒感染作为一个独立的因素增加少弱精子症的发生以及降低ICSI治疗周期的胚胎着床率和临床妊娠率（P〈0．05）。我们的研究结果提示了男性乙型肝炎病毒感染与较差的精子质量和ICSI治疗结局有关,而对IVF治疗结局没有明显的影响。     

Clinical management of infertile men with nonobstructive azoospermia

The clinical management of men with nonobstructive azoospermia (NOA) seeking fertility has been a challenge for andrologists, urologists, and reproductive medicine specialists alike. This review presents a personal perspective on the clinical management of NOA, including the lessons learned over 15 years dealing with this male infertility condition. A five-consecutive-step algorithm is proposed to manage such patients. First, a differential diagnosis of azoospermia is made to confirm/establish that NOA is due to spermatogenic failure. Second, genetic testing is carried out not only to detect the males in whom NOA is caused by microdeletions of the long arm of the Y chromosome, but also to counsel the affected patients about their chances of having success in sperm retrieval. Third, it is determined whether any intervention prior to a surgical retrieval attempt may be used to increase sperm production. Fourth, the most effective and efficient retrieval method is selected to search for testicular sperm. Lastly, state-of-art laboratory techniques are applied in the handling of retrieved gametes and cultivating the embryos resulting from sperm injections. A coordinated multidisciplinary effort is key to offer the best possible chance of achieving a biological offspring to males with NOA.     

Are conventional sperm morphology and motility assessments of predictive value in subfertile men?

Sperm morphology and motility are believed to be important prognostic factors for fertility. Results of a group of 67 men investigated for primary infertility who had mean sperm concentrations greater than 5 million per ml and who later produced pregnancies, were compared with those of 67 matched controls who remained infertile. All female partners were potentially fertile. The groups were matched for other known prognostic factors for fertility, namely, wife's age, the duration of infertility, sperm concentration and varicocele size. There were no significant differences between the two groups overall in the (mean +/- SEM)% of sperm with normal morphology (58.3 +/- 2.1; 58.5 +/- 2.2), or motility (40.6 +/- 1.8; 37.0 +/- 2.0). However, among oligospermic men from the two groups, sperm motility was significantly higher (P less than 0.05) in the subsequently fertile group (43.1 +/- 2.6%) than in the persistently infertile group (33.3 +/- 3.7). These results indicate that sperm morphology as currently assessed may not be important in predicting fertility in subfertile men with a mean sperm concentration over 5 million/ml and the % sperm motility may only be a relevant predictor in oligospermic men.     

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.     

Semen characteristics and inflammatory mediators in infertile men with different clinical diagnoses

This study was aimed at investigating whether semen characteristics in different clinical diagnoses of infertility are associated with PMN elastase, IL-6, IL-8, IL-1beta and TNFalpha levels detected in seminal plasma. Sixty-eight patients were divided into groups according to their clinical diagnosis: idiopathic infertility (group I), varicocele with infections (group II), varicocele (group III), infections (group IV), controls (group V). Physical examination and scrotal Eco-color Doppler was used to detect the varicocele. Patients with positive bacteriological semen analysis were considered as having an infection of the male reproductive tract. Samples were examined by light microscopy and transmission electron microscopy (TEM). TEM data were quantified with a mathematical formula furnishing a fertility index and the percentage of sperm apoptosis, immaturity and necrosis. PMN elastase/alpha1-PI complex levels were determined by ELISA and IL-6, IL-8, IL-1beta, TNFalpha by Bio-Plex Cytokine assay. Sperm concentration (I-II: p < 0.005; III-IV: p < 0.0001), motility (I-IV: p < 0.0001) and the fertility index (I: p < 0.005; II-IV: p < 0.0001) were significantly lower in the groups vs. controls, whereas sperm pathologies, except for apoptosis, were significantly higher in group I and apoptosis and necrosis were higher in group III. An increase in immaturity (p < 0.005) with a decrease in necrosis (p < 0.005) were observed in group III vs. group IV. Significantly higher levels of inflammatory mediators were detected in groups III and IV vs. controls. Despite a broad relationship among different inflammatory mediators, no correlation was found among them and the semen parameters, including indices from TEM analysis. In conclusion, patients with idiopathic infertility showed altered semen quality and normal levels of inflammatory mediators. Genitourinary infection and varicocele induced an inflammatory effect which could play a detrimental role in spermatogenesis, revealed by a decrease in sperm motility and the fertility index, concomitant with an increase in immaturity mainly in varicocele and necrosis in infection.     

Beta-endorphin in serum and seminal plasma in infertile men

Aim:To access beta-endorphin levels in serum as well as seminal plasma in different infertile male groups.Methods:Beta-endorphin was estimated in the serum and seminal plasma by enzyme-linked immunosorbent assay(ELISA)method in 80 infertile men equally divided into four groups:non-obstructive azoospermia(NOA),obstructive azoosper-mia(OA),congenital bilateral absent vas deferens(CBVAD)and asthenozoospermia.The results were compared tothose of 20 normozoospermic proven fertile men.Results:There was a decrease in the mean levels of beta-endorphin in the seminal plasma of all successive infertile groups(mean±SD:NOA 51.30±27.37,OA 51.88±9.47,CBAVD 20.36±13.39,asthenozoospermia 49.26±12.49 pg/mL,respectively)compared to the normozoospermicfertile control(87.23±29.55 pg/mL).This relation was not present in mean serum level of beta-endorphin betweenfour infertile groups(51.09±14.71,49.76±12.4,33.96±7.2,69.1±16.57 pg/mL,respectively)and the fertilecontrol group(49.26±31.32 pg/mL).The CBVAD group showed the lowest seminal plasma mean level of beta-endorphin.Testicular contribution of seminal beta-endorphin was estimated to be approximately 40%.Seminal beta-endorphin showed significant correlation with the sperm concentration(r=0.699,P=0.0188)and nonsignificantcorrelation with its serum level(r=0.375,P=0.185)or with the sperm motility percentage(r=0.470,P=0.899).Conclusion:The estimation of beta-endorphin alone is not conclusive to evaluate male reproduction as there aremany other opiates acting at the hypothalamic pituitary gonadal axis.(Asian J Androl 2006 Nov;8:709-712)     

Relationship between seminal ascorbic acid and sperm DNA integrity in infertile men

Ascorbic acid has recently been reported to protect sperm DNA from the damage induced by exogenous oxidative stress in vitro. But, there is no report on seminal ascorbic acid and sperm DNA fragmentation in infertile men. In this study, we asked whether sperm DNA damage correlates with seminal ascorbic acid levels. Sperm DNA fragmentation index (DFI) was analysed in 75 men by flow cytometry after acridine orange staining. We also measured the levels of seminal plasma ascorbic acid and total antioxidant capacity. Abnormal sperm DNA integrity (DFI >or= 30%) was observed in 12% of the patients with normal semen parameters and in 52% of the patients with abnormal semen parameters. There were significant correlations between the level of DFI and conventional semen parameters including sperm count, motility and morphology (r = -0.29, -0.55 and -0.53 respectively; p < 0.05). Seminal ascorbic acid level was significantly lower in the patients with leucospermia than the patient with normal semen parameters. Interestingly, a significantly greater percentage of men with abnormal DFI were observed in the patients with low levels of seminal ascorbic acid compared with those with normal or high levels of ascorbic acid (59% vs. 33%, p < 0.05). Men with insufficient seminal ascorbic acid frequently have sperm DNA damage.     

Clinical significance of reactive oxygen species in semen of infertile Indian men

Reactive oxygen species (ROS) levels in semen are believed to play both physiological and pathological roles in male fertility. The study was aimed to find the clinical significance of ROS levels in infertile Indian men. This pilot study included 33 idiopathic infertile men and 18 proven fertile controls. ROS levels in the washed sperm were measured using chemiluminescence assay and expressed as 10⁶ cpm per 20 million spermatozoa. Sperm count, percent sperm motility, and percent normal sperm morphology were found to be significantly ( P  < 0.0001) reduced in infertile men compared with the controls. Median (minimum, maximum range) ROS levels of the infertile group [24.90 (6.89, 44.71)] were found to be significantly ( P  < 0.0001) elevated compared with the fertile controls [0.167(0.15, 2.78)]. No significant correlation was seen between ROS levels and semen parameters. Elevated ROS levels in the idiopathic Indian infertile men may be one of the underlying reasons for impaired fertility. Therefore measurement of seminal ROS levels may be used in Indian infertile men for better understanding of the aetiology and selection of antioxidant regimen in the treatment of male infertility. However, large studies may be urgently warranted to find out the role of antioxidants in ROS elevated Indian infertile men through randomised, controlled clinical study.     

男性不育伴慢性病毒性肝炎患者血清和精液氧化应激水平及相关参数分析

目的:探讨男性不育伴慢性病毒性肝炎患者血清和精液丙二醛(MDA)、对氧磷酶-1(PON-1)水平及对精液相关参数的影响。方法:选择有生育力的男性健康对照组50例,男性不育组42例,男性不育伴慢性病毒性肝炎组45例,采用分光光度法测定血清和精浆MDA水平、PON-1活性,采用吖啶橙荧光染色法测定精子DNA碎片化指数(DFI)。结果:不育伴肝炎组血清和精浆MDA水平明显高于对照组和不育组(P<0.01和P<0.05),而PON-1活性明显低于对照组和不育组(P<0.01和P<0.05)。不育伴肝炎组精子活动率、精子存活率均明显低于对照组和不育组(P<0.01和P<0.05),而精子DFI明显高于对照组和不育组(P<0.01和P<0.05)。三组精子浓度无显著性差异(P>0.05);不育组与不育伴肝炎组精液WBC显著高于对照组(P<0.05)。男性不育伴慢性病毒性肝炎组精浆中的MDA水平、PON-1活性分别与血清中的MDA和PON-1呈显著正相关(r=0.57和0.48,P<0.01)。结论:病毒引起慢性活动性肝炎会使生殖系统氧化应激加强,加重对精子的损害,影响精子质量。