S.S. Allamaneni

Role of antioxidants in treatment of male infertility: an overview of the literature

Seminal oxidative stress in the male reproductive tract is known to result in peroxidative damage of the sperm plasma membrane and loss of its DNA integrity. Normally, a balance exists between concentrations of reactive oxygen species and antioxidant scavenging systems. One of the rational strategies to counteract the oxidative stress is to increase the scavenging capacity of seminal plasma. Numerous studies have evaluated the efficacy of antioxidants in male infertility. In this review, the results of different studies conducted have been analysed, and the evidence available to date is provided. It was found that although many clinical trials have demonstrated the beneficial effects of antioxidants in selected cases of male infertility, some studies failed to demonstrate the same benefit. The majority of the studies suffer from a lack of placebo-controlled, double-blind design, making it difficult to reach a definite conclusion. In addition, investigators have used different antioxidants in different combinations and dosages for varying durations. Pregnancy, the most relevant outcome parameter of fertility, was reported in only a few studies. Most studies failed to examine the effect of antioxidants on a specific group of infertile patients with high oxidative stress. Multicentre, double-blind studies with statistically accepted sample size are still needed to provide conclusive evidence on the benefit of antioxidants as a treatment modality for patients with male infertility.

Role of free radicals in female reproductive diseases and assisted reproduction

Infertility is a common problem experienced by many couples. Numerous treatments are available for female infertility. However, in some cases, the treatment is empirical in nature because the aetiology of infertility is not fully understood. Recently, reactive oxygen species (ROS) have been shown to have an important role in the normal functioning of reproductive system and in the pathogenesis of infertility in females. Reactive oxygen species may also play a role in other reproductive organ diseases of women such as endometriosis. Oxidative stress develops when there is an imbalance between the generation of ROS and the scavenging capacity of antioxidants in the reproductive tract. It affects both natural and assisted fertility. Because assisted reproductive techniques are used extensively in the treatment of infertility, it is critical to understand the in-vitro conditions that affect fertilization and embryo development. Treatments that reduce oxidative stress may help infertile women with diseases that are caused by this imbalance. Such strategies include identifying the source of excessive generation of ROS, treating the primary cause, and in-vitro and in-vivo supplementation of antioxidants. Research is in progress to identify the mechanisms that are involved in the aetiology of female reproductive diseases caused by ROS, and to create effective strategies that can counteract oxidative stress.

Chemiluminescence technique for measuring reactive oxygen species

Accurate assessment of reactive oxygen species (ROS) concentrations may help in the diagnosis of infertility. The chemiluminescence technique, which uses a luminometer to measure ROS, is a common method of assessment. A better understanding of the chemiluminescence technique will allow its proper application in reproductive medicine. A wide range of luminometers are available in the market, and laboratories should select the instruments that suit their individual needs.

Identification of male factor infertility using a novel semen quality score and reactive oxygen species levels.

PURPOSE To determine whether patients with male factor infertility can be accurately identified by calculating a novel semen quality score and measuring levels of reactive oxygen species during routine infertility screening. METHODS Semen samples from 133 patients and 91 healthy donors were evaluated with manual and computer-assisted semen analysis. A principal component analysis model was employed to calculate a semen quality score. In brief, this score was calculated by base 10 logarithms multiplied by varying weights given to 9 sperm parameters. Reactive oxygen species levels were measured using chemiluminescence assay. RESULTS The semen quality score had a sensitivity of 80.45% and accuracy of 77% at a cutoff of 93.1 in identifying patients with male factor infertility. The area under the receiver operating characteristic curves for the semen quality score was 84.28% (95% CI: 65.22%-100%). Reactive oxygen species levels [log10 (reactive oxygen species +1)] were significantly higher in male factor infertility patients. Reactive oxygen species had a sensitivity of 83.47% and specificity of 60.52% with an accuracy of 75% at a cutoff of 1.25 in identifying male factor infertility patients. The area under the receiver operating characteristic curve for reactive oxygen species levels was 78.92% (95% CI: 72.60%-85.23%). Semen quality scores were significantly and negatively correlated with reactive oxygen species levels in the donors and the male factor infertility patients. CONCLUSIONS The semen quality score and reactive oxygen species levels in semen samples appear to be strongly associated with male factor infertility. Because both of these parameters are more sensitive than individual sperm parameters in identifying male factor infertility, they should be included in routine infertility screening.

1363: Are Standard Values of Semen Analysis by World Health Organization Accurate?

Introduction and Objective: Semen analysis is the most important initial diagnostic test available for the evaluation of male infertility. Sperm concentration and the number of motile and morphologically normal spermatozoa are significant factors influencing in vivo and in vitro pregnancy. The objective of our study was to evaluate if current normal values of semen analysis as described by World Health Organization (WHO) are accurate in diagnosing male infertility. Methods: Semen samples were collected from proven fertile donors (n = 47) and patients attending infertility evaluation (n = 406). Routine semen analysis was performed according to WHO guidelines and morphology was also assessed by Tygerberg’s strict criteria. ROC curve analysis was used to calculate the areas under curve (AUC), sensitivity and specificity at 2 different cut-off points, 1) standard WHO cut-off values; 2) cutoff values provided by statistical software (StatsDirect Ltd., Gresham Way, UK) by giving equal weightage to sensitivity and specificity of sperm parameters. Results: Significant overlapping of sperm characteristics was seen between proven fertile donors and patients attending infertility evaluation. Most of the proven fertile men and patients attending infertility evaluation had lower percentage of morphologically normal spermatozoa, however they had normal sperm concentration and motility according to WHO standards. Sperm morphology by WHO and Tygerberg’s strict criteria had higher AUC with better sensitivity and specificity compared to sperm concentration and motility at the current WHO cut-off values. A further decrease in the cut-off value of morphology significantly helped in improving specificity thus overall accuracy (Table). Conclusions: Sperm morphology by WHO and Tygerberg’s strict criteria are a better discriminator of fertile men and patients evaluated for infertility evaluation than sperm motility and concentration. Lowering the cut-off values of morphology can further improve accuracy in discriminating the two groups. Print this Page for Your Records Close Window

1583: Identifying Infertile Men with Male-Factor Infertility with a Novel Semen Quality (SQ) Score

Introduction and Objective: Semen analysis constitutes the main component in the evaluation of male infertility. Semen Quality (SQ) score has been demonstrated as a better predictor of semen quality compared to the individual characteristics of sperm. The purpose of this study was to determine whether infertile patients with male-factor infertility (MFI) could be accurately identified using a well-defined SQ score. Methods: The study included results from 124 MFI patients. Healthy male volunteers with normal semen parameters (n = 70), according to World Health Organization guidelines, were used as controls. Manual semen analysis was performed and sperm motion kinetics were assessed using a computer assisted semen analyzer (CASA, IVOS, 10.7s, Hamilton Thorne Research, Beverly, Mass). Principal component analysis model was used to calculate SQ score that accounts for most of the variability observed among the battery of interrelated semen variables. Base 10 logarithms of nine sperm parameters (concentration, motility, sperm morphology according to WHO guidelines and Tygerbergs strict criteria, VCL, VSL, VAP, LIN, and ALH) were used. Results: In donors, the mean and standard deviation of SQ score was 99.39 ± 9 compared to 75.57 ± 18.55 in MFI patients (P< 0.0001). Amongst the patient population, 93% of men had SQ score <100, while 7% (9/133) had a SQ score >100. SQ score had a sensitivity of >93% when a cutoff of 100 was used, i.e. SQ score was able to correctly identify 93% of the patients as being MFI patients. The overall accuracy in identifying the donors and patients was 77%. Using a cutoff value of 95 and 90, the accuracy was 79% and 79% but there was a decrease in sensitivity. Conclusions: SQ score derived from patient’s semen analysis may be an easy and effective tool for clinicians in the initial evaluation of patients being examined for their infertility problem. Various predictors of semen quality in donors and MFI patients using different cutoff values of SQ

1386: Role of Seminal Growth Factors in Infertile Men with Varicocele

Introduction and Objective: Growth factors such as basic fibroblast growth factor (bFGF) and transforming growth factors (TGF-β1 and β2) are polypeptides that function as key regulatory molecules affecting germ cell proliferation and differentiation. The pathophysiology by which varicocele produces sperm dysfunction and infertility is not completely understood. The objective of our study was to examine the relationship of seminal growth factors with normal and abnormal semen parameters in infertile men with varicocele. Methods: In our prospective-controlled study, a group of infertile men with clinical varicocele (varicocele patients with normal semen parameters: n = 9; and varicocele patients with abnormal semen parameters: n = 16) were selected. Fifteen healthy donors with normal standard semen parameters served as a control. Sperm concentration, motility, and morphology were assessed according to WHO guidelines. Levels of growth factors (bFGF, TGF-β1 and TGF-β2) were measured in seminal plasma using ELISA assay kits (Quantikine, R & D Systems, Minneapolis, MN). The sensitivity of the assay was: bFGF <3pg/mL; TGF-β1 and β2 <7pg/mL. Results: Significant differences were seen in sperm concentration (P< 0.0001), motility (P< 0.0003), and morphology (P< 0.0001) in men with varicocele compared to controls. Levels of TGF-β1 were significantly higher in varicocele patients with abnormal semen parameters (P = 0.03) compared to controls (Table 1). In addition, the TGF-β1 levels in these patients correlated negatively with sperm motility (r = -0.57, P = 0.02) and morphology (r = -0.57, P = 0.02). Levels of TGF-β2 and bFGF were comparable between varicocele patients and control groups. Conclusions: TGF-β1 may play an indirect role in the pathophysiology of infertility in varicocele patients. The correlation of these growth factors with poor quality semen parameters in these patients may reflect dysfunctional spermatogenesis. Values of different growth factors in infertile men with varicocele and in healthy controls

Disturbances in gonadal axis in women with anorexia nervosa

UNLABELLED Anorexia nervosa negatively affects multiple body systems including the reproductive system. AIM To assess the disturbances in the hypothalamic-pituitary-gonadal axis (HPG) and the relationship between the gonadotropins and body weight, duration of the disease and amenorrhea we studied 40 female anorexic patients (aged 14-31 years) with a body mass index (BMI) 15.14+/-1.80 kg/m(2) and a degree of weight loss 28.67+/-8.74%. Fifteen healthy, age-matched women with normal weight served as controls. METHODS We investigated the disturbances in the gonadotropin levels before and after stimulation with gonadotropin-releasing hormone (GnRH) 100 microg i.v. One week later 100 mg of clomiphene citrate (CC) was administered orally for 5 days. RESULTS Basal levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were significantly lower in the patients. The responses of LH to GnRH were diminished, but those of FSH were exaggerated. However, after clomiphene citrate administration, LH increased 5.4 times whereas FSH increased 1.7 times. The basal levels of LH were significantly correlated with body weight (r=+0.373, p<0.05), BMI (r=+0.385, p<0.01) and percentage of the weight loss (r=-0.356, p<0.05). FSH levels were positively correlated with the duration of the disease (r=+0.481, p<0.01) and amenorrhea (r=+0.540, p<0.01). CONCLUSIONS Our study demonstrates dissociation in the secretion of gonadotropins after hypothalamic stimulation in anorexic patients. It also reveals the relationship between alterations in the hormones of the HPG axis, not only with the changes in body weight, but also with the duration of the disease.