Adel Zalata

MedCalc: a new computer program for medical statistics

In recent years, the use and abuse of statistics in the medical literature has extensively been reviewed. Amongst others, the importance of the P-value has been challenged and the use of misleading graphics, including 3-dimensional displays, has been criticized. The ease of access to more complex statistical procedures, since the introduction of several statistical software packages for personal computers, has been identified as one of the factors involved in the misuse of statistics. Therefore, we have developed a new computer program that includes those statistical procedures commonly encountered in the medical literature and in statistical textbooks for medical researchers. More complex statistical analyses are not implemented in the software. If researchers with limited statistical training require more sophisticated statistical analyses, they should refer to a statistician, not to a more complete statistical software package.

Evaluation of the role of reactive oxygen species in male infertility

Reactive oxygen species (ROS) production from spermatozoa has been measured by chemiluminescence in the two fractions of a Percoll gradient column (47 and 90%). Chemiluminescent signals were recorded in each fraction after the addition of luminol and horse-radish peroxidase (basal state), and after stimulation with formyl-methionyl-leucyl-phenylalanine and phorbol ester (PMA). Oligozoospermic samples show a higher rate of ROS production than the normozoospermic samples in both fractions of Percoll. Also, ROS were generated at a higher rate by asthenozoospermic samples in the 90% Percoll fraction than by normal samples after stimulation with PMA. Our data confirm that fact that white blood cells play a major role in the production of ROS, even after purification on a Percoll gradient. Immunological cases were also found to be associated with an increased production of ROS, which may be caused by the same underlying pathological condition responsible for the production of the antibodies. Repeated centrifugation of the samples triggers a burst of ROS in excess of that produced after Percoll preparation. In addition, superoxide dismutase activity was found to be significantly increased in cases with an elevated production of ROS. It is concluded that measuring the ROS generation by semen may yield useful information on the functional capacity of spermatozoa, which may be used to improve the success of male infertility management.

Mechanisms of sperm deficiency in male accessory gland infection.

Summary. The presence of 2 million or more peroxidase‐positive white blood cells per ml of semen, or the diagnosis of male accessory gland infection, is associated with important biochemical and biological changes in semen plasma and in the spermatozoa, reducing their fertilizing potential in vitro and in vivo (e.g., during intra‐uterine insemination). In addition to the effects of reactive oxygen species, and its influence on the essential fatty acid composition of the sperm membrane, potentially unfavourable effects can occur through the intermediate of increased concentrations of certain cytokines, and decreased activity of enzymes such as alpha‐glucosidase.

Follicle-stimulating hormone receptor polymorphism and seminal anti-Mullerian hormone in fertile and infertile men

Follicle‐stimulating hormone (FSH) is fundamental for Sertoli cell function stimulating spermatogenesis and follicular growth by a specific receptor (FSHR). This work aimed to investigate the occurrence of Asn and Ser FSHR gene variants and its relationship with seminal anti‐Müllerian hormone (AMH) among normozoospermic and infertile oligoasthenozoospermic (OAT) males. Eighty‐two Caucasian males grouped into normozoospermic healthy controls (n = 30) and infertile OAT males (n = 52). FSHR gene variants were determined by DNA from anti‐coagulated blood and underwent polymerase chain reaction (PCR) amplification and electrophoresis in detecting amplification products. AMH in seminal plasma was determined by ELISA. The results showed that the frequency of FSHR gene variants among fertile men was 46.7% Asn/Asn (N680S), 33.3% Asn/Ser, and 20% Ser/Ser, whereas among OAT men were 34.6%, 38.5% and 26.9% respectively with nonsignificant differences. Seminal AMH was significantly higher in fertile than infertile OAT men. There was significant increase in seminal AMH with Asn/Asn variant of FSHR gene than those with Asn/Ser or Ser/Ser. It is concluded that FSH gene variants showed no difference in distribution between fertile or infertile OAT men. However, when correlated with seminal AMH values, there was an increase in Asn/Asn in men with high seminal AMH.

Seminal clusterin gene expression associated with seminal variables in fertile and infertile men.

PURPOSE CLU is a disulfide linked, heterodimeric protein associated with the clearance of cellular debris and apoptosis. We assessed the association of seminal CLU gene expression with seminal variables in fertile and infertile men. MATERIALS AND METHODS A total of 124 men were divided into healthy, fertile men with normozoospermia, and men with asthenozoospermia, asthenoteratozoospermia and oligoasthenoteratozoospermia. History was obtained, and clinical examination and semen analysis were done. In semen we assessed sperm acrosin activity, sperm DNA fragmentation and seminal CLU gene expression. RESULTS CLU RNA and CLU protein gene expression were significantly increased in semen samples of infertile men with oligoasthenoteratozoospermia > asthenoteratozoospermia > asthenozoospermia compared with healthy, fertile controls. CLU gene expression significantly correlated negatively with sperm count, motility, acrosin activity index, linearity index and linear velocity, and significantly correlated positively with the percent of sperm abnormal forms and DNA fragmentation. CONCLUSION CLU gene expression was significantly increased in the semen samples of infertile men. It correlated negatively with sperm count, motility, acrosin activity, linearity index and linear velocity, and positively with the percent of sperm abnormal forms and DNA fragmentation.

Cholesterol and desmosterol in two sperm populations separated on Sil-Select gradient

Sperm lipids are important for sperm viability, maturity and function. This study aimed to identify cholesterol and desmosterol composition of human spermatozoa of two sperm populations separated on Sil-Select gradient. Forty-eight males were divided into four groups namely healthy men (n = 13), asthenozoospermia (n = 11), asthenoteratozoospermia (n = 10) and oligoasthenoteratozoospermia (n = 14). Sperm cholesterol and desmosterol were estimated in two human sperm population separated by centrifugation in a discontinuous Sil-Select gradient. The results showed that cholesterol and desmosterol were the major sterols in human spermatozoa. Spermatozoa recovered from upper/lower layer interface (fraction I) had low fertilization potential, while those from the base (fraction II) had high fertilization potential. Median values of cholesterol and desmosterol in fraction I were 2.55 micromol and 0.77 micromol/10(9) spermatozoa and in fraction II were 1.16 micromol and 0.27 micromol/10(9) spermatozoa. Cholesterol/desmosterol ratio was significantly higher in fraction II than I (4.8 vs. 3.2, p < 0.01). Cholesterol, desmosterol, total phospholipids and sterols/phospholipids were negatively correlated with sperm concentrations, sperm motility, linear velocity, normal sperm morphology and acrosome reaction percentage whereas cholesterol/desmosterol ratio was positively correlated with these parameters. It is concluded that the difference in sterol composition of sperm subpopulations separated on Sil-Select gradient suggests that composition of sterols is related to sperm functions.

Long-term ofloxacin testicular toxicity: an experimental study

The aim of this study was to assess the long‐term toxic effect of ofloxacin on the testes and epididymides of 72 adult male albino rats. The rats were divided into group A and group B. Group A, which received ofloxacin for 14 days, was subdivided into two subgroups; LD‐14 received low dose 72 mg KBW−1 daily and HD‐14 received high dose 216 mg KBW−1 daily. Group B, which received ofloxacin for 28 days, was subdivided into two subgroups; LD‐28 received 72 mg KBW−1 and HD‐28 received 216 mg KBW−1 daily. Two matched control groups were followed up for 14 and 28 days respectively. The animals were evaluated for body weight, testicular weight, relative testicular weight, serum testosterone (T), epididymal sperm analysis (sperm count, motility, morphology, curvilinear velocity, linear velocity and linearity index) and testicular histopathology. The adverse effects of ofloxacin were correlated with increased treatment duration and/or dose. It is concluded that long‐term ofloxacin has a direct detrimental effect on the testicles of albino rats at the studied doses and durations.

Androgen Receptor Expression Relationship with Semen Variables in Infertile Men with Varicocele

PURPOSE Androgen receptor, a member of the nuclear receptor superfamily, has important roles in male reproductive function. It is required for sexual differentiation, pubertal development, spermatogenesis regulation, meiosis completion and spermatocyte transition to haploid round spermatids. We assessed the association of androgen receptor expression and semen variables in infertile men with varicocele. MATERIALS AND METHODS A total of 299 men were grouped into healthy, fertile controls, infertile men without varicocele and men with infertility associated with varicocele. A history was obtained, clinical examination and semen analysis were done and reproductive hormones were estimated. Androgen receptor expression and the acrosome reaction were determined in recovered spermatozoa. RESULTS Androgen receptor expression was significantly decreased in infertile men with varicocele more than in infertile men without varicocele compared to fertile controls. Androgen receptor correlated positively with sperm count, motility, normal forms, velocity, linear velocity, acrosome reaction and α-glucosidase. It correlated negatively with serum follicle-stimulating hormone and estradiol. Multiple stepwise regression analysis of androgen receptor expression revealed that the sperm acrosome reaction and linearity index were the most affected independent variables. CONCLUSIONS Androgen receptor expression was significantly decreased in infertile men with varicocele more than in infertile men without varicocele compared to fertile men. Androgen receptor expression correlated positively with sperm count, motility, normal forms, velocity, linear velocity and acrosome reaction.

Tumor necrosis factor-α gene polymorphism relationship to seminal variables in infertile men.

OBJECTIVE To assess the tumor necrosis factor (TNF)-α gene polymorphism relationship with seminal variables in fertile men (N) and those with asthenozoospermia (A), asthenoteratozoospermia (AT), and oligoasthenoteratozoospermia (OAT). MATERIALS AND METHODS A total of 50 infertile men without a female factor who were attending a fertility clinic and 48 fertile men were randomly screened for semen analysis, analysis of the TNF-α promoter region for polymorphism, seminal caspase-9, acrosin activity, α-glucosidase, and reproductive hormones. RESULTS The TNF-α GG genotype was present in 83.9%, 72.7%, 66.7%, and 59.5%, the TNF-α AA genotype in 3.2%, 6.8%, 10.4%, and 11.9%, and TNF-α AG genotype in 12.9%, 20.5%, 22.9%, and 28.6% in the N, A, AT, OAT groups, respectively. The occurrence of A allele was significantly greater among infertile patients than among fertile controls (21.6% vs 9.7%; odds ratio 0.388, 95% confidence interval 0.2 to 0.75, P = .005). Men with the TNF-α AA genotype demonstrated a significant decrease in the sperm count, sperm motility, normal sperm morphology, acrosin activity, and seminal α-glucosidase and a significant increase in seminal caspase-9 compared with those with the TNF-α GG genotype. CONCLUSION This single nucleotide polymorphism in the TNF-α(-308) gene was associated with significantly increased seminal caspase-9 and a significantly decreased sperm count, sperm motility, normal sperm morphology, acrosin activity, and seminal α-glucosidase.

ACE Gene Insertion/Deletion Polymorphism Seminal Associations in Infertile Men

PURPOSE We assessed seminal associations of the ACE gene insertion/deletion polymorphism in infertile men. MATERIALS AND METHODS A total of 405 men were investigated, divided into healthy fertile men, and those with asthenozoospermia, asthenoteratozoospermia and oligoasthenoteratozoospermia, respectively. They underwent semen analysis, and assessment of sperm acrosin activity, hypo-osmotic swelling, seminal 8-iso-prostaglandin-F(2α), total antioxidant capacity, α-glucosidase and ACE gene polymorphisms. RESULT The ACE insertion/insertion genotype was noted in 182 men, including 76.5% of healthy fertile men, and 47.4%, 39.8% and 17.6% of those with asthenozoospermia, asthenoteratozoospermia and oligoasthenoteratozoospermia, respectively. The ACE insertion/deletion genotype was noted in 133 men, including 13.7% of healthy fertile men, and 42.3%, 27.5% and 47.2% of those with asthenozoospermia, asthenoteratozoospermia and oligoasthenoteratozoospermia, respectively. The ACE deletion/deletion genotype was identified in 90 men, including 9.8% of healthy fertile men, 10.3%, 32.70% and 35.2% of those with asthenozoospermia, asthenoteratozoospermia and oligoasthenoteratozoospermia, respectively. Men with the ACE deletion/deletion and insertion/deletion genotypes showed a significant decrease in sperm count, motility, linear velocity and normal forms, acrosin activity index, hypo-osmotic swelling test and seminal α-glucosidase, and significantly increased seminal 8-iso-prostaglandin-F(2α) than those with the ACE insertion/insertion genotype. CONCLUSIONS ACE gene deletion polymorphism is associated with abnormal seminal variables, such that carriers of the ACE deletion/deletion genotype have higher seminal oxidative stress.