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.

Follicle-stimulating hormone receptor polymorphism and seminal anti-Müllerian 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.

The effects of combined conventional treatment, oral antioxidants and essential fatty acids on sperm biology in subfertile men.

We evaluated the effects of combined conventional treatment, oral antioxidants (N-acetyl-cysteine or vitamins A plus E) and essential fatty acids (FA) on sperm biology in an open prospective study including 27 infertile men. The evaluation included sperm characteristics, seminal reactive oxygen species (ROS), FA of sperm membrane phospholipids, sperm oxidized DNA (8-OH-dG), and induced acrosome reaction (AR). Treatment did not improve sperm motility and morphology, nor decrease the concentration of round cells and white blood cells in semen. Sperm concentration increased in oligozoospermic men (7.4+/-1.3 to 12.5+/-1.9 million/ml). Treatment significantly reduced ROS (mean+/-SEM) (775.3+/-372.2 to 150.3+/-105.2 x 10(3)counts/10 second) and 8-OH-dG (45.3+/-10.4 to 16. 8+/-3.3 fmol/microg DNA). Treatment increased the AR (55.1+/-2.2 to 71.6+/-2.2%), the proportion of polyunsaturated FA of the phospholipids, and sperm membrane fluidity. The overall pregnancy rate was 4.5% in 134 months. The per month pregnancy rate tended to be higher in partners of (ex)-smokers (7.15%, n=14,70 months) than in never-smokers (1.6%, n=13,64 months) (OR:4.57, 95% Cl:0.55-38.1).

Mechanisms and effects of male genital tract infection on sperm quality and fertilizing potential: the andrologist's viewpoint.

There are several mechanisms acting in synergism that can impair sperm characteristics of patients with accessory gland infection. In some cases, conventional sperm variables are disturbed with oligo and/or asthenozoospermia. In other patients, these sperm variables may appear normal, but the functional capacity of spermatozoa may be impaired. In particular, changes in the composition of the sperm membrane may result in reduced acrosome reactivity and capacity to fuse with the oolemma, and oxidative damage of the sperm DNA may induce mutagenesis. Changes in the biochemical make-up of seminal plasma can also reduce the in-vivo fertilizing capacity of spermatozoa, and infection-related disruption of the blood-testis barrier can induce the generation of anti-sperm antibodies and immunological infertility. Many of these functional abnormalities will not become evident upon 'basic semen analysis', which explains why some authors are unable to link infection of the accessory sex glands to subfertility. Also, functional and anatomical damage acquired as a result of infection is often permanent and not reversible by (antibiotic) treatment. Clearly, there are many more aspects of male accessory gland infection that require investigation. Available data should stimulate clinicians to place more emphasis on the prevention of infection-related infertility than on its treatment, as the latter is often unsuccessful.

The correlates and alleged biochemical background of the resazurin reduction test in semen.

Reduction of the blue dye resazurin to pink resorufin is used to estimate the concentration of metabolically active spermatozoa in semen samples. In order to quantify the reduction of resazurin, a spectrophotometric method was developed measuring the change from blue to pink in the butanol extracted colour. The biochemical mechanisms involved in the reduction of resazurin by motile spermatozoa and seminal plasma were investigated. Addition of NADH + H+ to sperm suspension or seminal plasma increased the reduction of resazurin. The reduction reaction was inhibited by high concentrations of dicoumarol, a specific inhibitor of the diaphorase enzyme, in a dose-dependent manner. It is suggested that the sperm diaphorase enzyme transfers electrons from NADH + H+ to resazurin, reducing it to resorufin. The degree of resazurin reduction was strongly correlated with the concentration of motile spermatozoa recovered from the 90% Percoll fraction (r = 0.98, p < 0.001). A positive correlation was also found between the reducing capacity of seminal plasma (n = 62) on the one hand, and sperm concentration (r = 0.72, p < 0.0001), progressive motility (r = 0.45, p < 0.01), normal morphology (r = 0.50, p < 0.01), and gamma-glutamyltransferase (r = 0.36, p < 0.05) on the other hand. These findings, together with our previous observations that the reduction reaction is inhibited by reactive oxygen species and polymorphonuclear white blood cells, increase our understanding of the biochemical basis of the resazurin test and may provide better insight into the interpretation of this test.

White blood cells cause oxidative damage to the fatty acid composition of phospholipids of human spermatozoa.

The lipid composition of the sperm membrane has been shown to exert a significant effect upon the functional quality of spermatozoa. We have studied the effect of induced peroxidation and of the presence of polymorphonuclear white blood cells (WBCs) on the fatty acid composition of the phospholipids of human spermatozoa. The spermatozoa were fractionated by a discontinuous Percoll gradient in two fractions (47% and 90% Percoll). Induced peroxidation of spermatozoa was assessed by determining the production of thiobarbituric acid reactive substances (TBARS), mostly malondialdehyde, after incubation with ferrous sulphate and sodium ascorbate as a promoter of peroxidation. TBARS production after induction of peroxidation was correlated with the abundance of polyunsaturated fatty acids (PUFA)(r = 0.68, p < 0.0001), with the double bond index (r = 0.72, p < 0.0001), and with the oxidative potential index (r = 0.73, p < 0.0001) of fatty acids of phospholipids. In comparison with samples containing > 1 x 10(6) WBCs/mL, those with < 1 x 10(6) WBCs/mL contained higher proportions of PUFA (90% Percoll, p < 0.05; 47% Percoll, p < 0.05), total omega 3 fatty acids (90% Percoll, p < 0.05; 47% Percoll, p < 0.001), docosahexaenoic acid (90% Percoll p < 0.05; 47% Percoll, p < 0.05), and double bond index (90% Percoll, p < 0.05; 47% Percoll, p < 0.001). In addition, mean melting point was significantly lower (90% Percoll, p < 0.05; 47% Percoll, p < 0.001) in samples with < 1 x 10(6) WBCs, indicating higher membrane fluidity. The increase of TBARS production by spermatozoa after incubation with the xanthine-xanthine oxidase system and/or ferrous sulphate as promoter of peroxidation was associated with a significant decrease of PUFA. Incubation of spermatozoa with WBCs, with or without activation by phorbol ester, decreased the PUFA (p < 0.05). Also, TBARS production was increased (p < 0.01) after activation of WBCs with phorbol ester. Our data provide evidence that oxidative stress induced by WBCs has a damaging effect on the polyunsaturated fatty acids of sperm phospholipids which may result, amongst other effects, in decreased membrane fluidity.

The fatty acid composition of phospholipids of spermatozoa from infertile patients.

The lipid composition of the sperm membrane has a significant effect upon the functional characteristics of spermatozoa. In the present study we investigated the fatty acid (FA) composition of subpopulations of spermatozoa separated on a discontinuous Percoll gradient (47:90%) and the FA composition of phospholipids (PL) of sperm heads and tails in both normal and abnormal semen samples. In normozoospermic samples, polyunsaturated fatty acids (PUFA) represented 34.0 +/- 1.3 (mean +/- SE, mole %) and 25.6 +/- 1.2% of total FA of PL of the 47 and 90% Percoll fractions respectively. Docosahexaenoic acid (22:6omega3, DHA) contributed to more than 60% of total PUFA. DHA was significantly lower in both the 47% (P < 0.05) and the 90% (P < 0.01) Percoll fractions of oligozoospermic samples and in the 90% Percoll layer of asthenozoospermic samples (P < 0.01), compared with normozoospermic samples. The omega6/omega3 ratio was significantly increased in both Percoll fractions of samples with oligozoospermia (47%, P < 0.001 and 90%, P < 0.001) or with asthenozoospermia (47%, P < 0.05 and 90%, P < 0.001) compared with normozoospermic samples. The oxidative potential index (OPI) of spermatozoa recovered from the 47% Percoll layer was significantly higher (P < 0.0001) than of those recovered from the 90% Percoll. Mean melting point (MMP), an index of membrane fluidity, was significantly lower in head than in tails (P < 0.01) of spermatozoa, and also in both the 47% (P < 0.01) and 90% (P < 0.001) Percoll fractions of normozoospermic samples in comparison with oligozoospermic samples. The MMP was significantly higher (P < 0.05) in samples of patients with idiopathic oligo/asthenozoospermia, varicocele, and male accessory gland infection (MAGI). These differences in FA composition of PL in subpopulations of human spermatozoa, and in their heads and tails may be related to sperm maturity and to differences in physiological function.