In Vitro Effects of Metal Ions (Fe2+, Mn2+, Pb2+) on Sperm Motility and Lipid Peroxidation in Human Semen

The effects of divalent manganese ion (Mn²⁺), ferrous iron (Fe²⁺), and lead ion (Pb²⁺) on human sperm motility and lipid peroxidation were examined. Human semen from healthy male volunteers was incubated with 0, 5, 50, or 500 ppm divalent metal ions, and the sperm motility was determined at 0, 2, 4, 6, or 8 h by microscopy. Malondialdehyde (MDA) levels in seminal plasma was measured by high-performance liquid chromatography after 8 h of exposure. The results showed that 500 ppm Mn²⁺ or Pb²⁺ significantly inhibited sperm motility without an accompanying change in seminal MDA levels. Incubation with Fe²⁺ significantly inhibited sperm motility at 5 ppm, associated with a marked rise in MDA levels. Our results suggested that Fe²⁺ may induce lipid peroxidation to inhibit sperm motility. In the case of Mn²⁺ and Pb²⁺ there is an absence of seminal lipid peroxidation and the observed inhibition of sperm motility at high concentrations is not biologically or environmentally relevant.     

Assessment of semen function and lipid peroxidation among lead exposed men

The study population included healthy, fertile men, employees of Zinc and Lead Metalworks (n=63). Workers exposed to lead were divided into two groups: a group with moderate exposure to lead (ME) - blood lead level (PbB) 25-40 microg/dl and a group with high exposure to lead (HE) PbB=40-81 microg/dl. The control group consisted of office workers with no history of occupational exposure to lead. Evaluation of lead, cadmium and zinc level in blood and seminal plasma, zinc protoporphyrin in blood (ZPP), 5-aminolevulinic acid in urine (ALA), malondialdehyde (MDA) in seminal plasma and sperm analysis were performed. No differences were noted in the concentration of cadmium and zinc in blood and seminal plasma in the study population. Lipid peroxidation in seminal plasma, represented as MDA concentration, significantly increased by about 56% in the HE group and the percentage of motile sperm cells after 1 h decreased by about 34% in comparison to the control group. No statistically significant correlation between other parameters of sperm analysis and lead exposure parameters nor between lead, cadmium and zinc concentration in blood and seminal plasma were found. A positive association between lead intoxication parameters (PbB, ZPP, lead seminal plasma) and MDA concentration in sperm plasma and inverse correlation with sperm cells motility (PbB, ZPP) was found. An increased concentration of MDA was accompanied by a drop in sperm cells motility. In conclusion, we report that high exposure to lead causes a decrease of sperm motility in men most likely as a result of increased lipid peroxidation, especially if the level in the blood surpasses the concentration of 40 microg/dl.     

The combined effect of Pb2+ and Mn2+ on monoamine uptake and Na+, K+-ATPase in striatal synaptosomes

Rat striatal synaptosomes (P2-fraction) were subjected to lipoperoxidation by the addition of 120 microM Fe2+ and 200 microM ascorbic acid. This preparation (pretreated synaptosomes) was used to investigate the interaction of Pb2+ and Mn2+ on the uptake of tritiated catecholamines, Na+, K+-ATPase activity and malondialdehyde (MDA) formation in order to understand the mechanism of enhanced neurotoxicity by concurrent exposure to these metals. The combination of Pb2+ and Mn2+ (25 microM + 100 microM, respectively) produced a significant increase in the uptake of 3H-Dopamine only in the untreated synaptosomes. No significant effect was noted on the uptake of 3H-Norepinephrine in either pretreated or untreated synaptosomes. However, the combination of Pb2+ and Mn2+ produced a pronounced decrease in the activity of Na+, K+-ATPase, but the magnitude of the change was the sum of the individual metal effects. Metal interaction did not produce any significant change in the formation of MDA compared to the control (without addition of metals). These results indicate that Pb2+ and Mn2+ interaction may produce inhibition in the activity of transport ATPase in both the preparation of synaptosomes, with more pronounced effect of synaptosomes subjected to lipoperoxidation and these changes may be responsible for the disruption in the physiology of nerve impulse transmission.     

Evaluation of iron binding and peroxide-mediated toxicity in rat hepatocytes.

A novel assay was developed to determine subnanomolar amounts of Fenton-reactive iron (FRI) in biological tissues. FRI represents that pool of iron that is redox active and capable of participating in a model Fenton reaction. The FRI was used to identify a kinetically-defined cellular iron binding site. This site displays positive cooperativity, with apparent kinetic constants of Kd = 10.6 microM, Bmax = 20.7 nmol/mg protein, and the Hill coefficient = 1.4. After addition of exogenous ferrous ammonium sulfate to hepatocytes, binding occurred within a few seconds and was stable for at least an hour. Free extracellular iron, but not bound iron, stimulated lipid peroxidation in hepatocytes. In contrast, bound but not free iron produced a concentration-dependent increase in tert-butyl hydroperoxide (TBH)-mediated toxicity, suggesting the toxicological relevance of bound, rather than free iron. Furthermore, the hydroxyl radical scavengers mannitol and 2-deoxyribose inhibited Fe2/TBH-mediated lipid peroxidation, but not cell killing, suggesting that hydroxyl radical may not be involved in the critical toxic event. The divalent cations Mn2+ and Co2+ inhibited iron-mediated hepatocyte killing in the presence of TBH, but only if added prior to Fe2+. Mn2+, but not Co2+, inhibited Fe(2+)-mediated lipid peroxidation regardless of the order of addition. These results indicate the existence of a specific, kinetically-defined cellular iron binding site. Such binding is involved in peroxide-mediated toxicity, but independent of lipid peroxidation. The specific nature of this site and involvement with other forms of chemical intoxication or cellular iron homeostasis are unknown.     

Inhibitory effects of melatonin on ferric nitrilotriacetate-induced lipid peroxidation and oxidative DNA damage in the rat kidney

Ferric nitrilotriacetate (Fe-NTA) is a known complete renal carcinogen which induces lipid peroxidation and oxidative DNA damage in rat kidney. In this study, the in vivo and in vitro effects of melatonin on Fe-NTA-induced lipid and oxidative DNA damage were determined. The levels of malondialdehyde (MDA) and 4-hydroxyalkenals (4-HDA) were assayed as an index of lipid peroxidation and the levels of 8-hydroxydeoxyguanosine (8-OH-dG) as an endpoint of oxidative DNA damage. In in vitro studies, the increased levels of MDA and 4-HDA induced by Fe-NTA were observed to be dose-dependent and time-dependent. The increase in lipid peroxidation was inhibited by melatonin in a concentration-dependent manner. When Fe-NTA(15 mg Fe/kg body weight) was intraperitoneally injected into rats, the levels of MDA + 4-HDA and 8-OH-dG in the rat kidney were increased 1 h after its administration as compared to levels of these constituents in the control group. Pretreatment with melatonin (25 mg/kg or 50 mg/kg) 30 min before the Fe-NTA injection resulted in a significant reduction in the levels of lipid peroxidation and 8-OH-dG induced by Fe-NTA in the rat kidney. These results are consistent with the conclusion that the toxicity of Fe-NTA is due to the generation of reactive oxygen species and that melatonin's protective effects relate to its direct radical scavenging ability and due to other antioxidative processes induced by the indole.     

The effect of methoxychlor on the epididymal antioxidant system of adult rats

Methoxychlor is widely used as a pesticide in many countries and has been shown to induce reproductive abnormalities in male rats, causing reduced fertility. The mechanism of action of methoxychlor on the male reproductive system is not clear. In the present study we investigated whether administration of methoxychlor induces oxidative stress in the epididymis and epididymal sperm of adult rats. Methoxychlor (50, 100, or 200 mg/kg body weight/day) was administered orally for 1, 4, or 7 days. The animals were killed using anesthetic ether 24 h after of the last treatment. Epididymal sperm were collected by cutting the epididymis into small pieces in Ham's F-12 medium at 35 degrees C. The body weight and weights of the testis, liver, and kidney did not show any significant changes in the methoxychlor-treated rats. The weight of the epididymis, seminal vesicles, and ventral prostate as well as epididymal sperm counts decreased after 50, 100, or 200 mg/kg/day for 7 days but remained unchanged after shorter courses of treatment. Epididymal sperm motility was decreased in a dose-dependent manner in the animals treated with methoxychlor for 4 or 7 days. The activities of the antioxidant enzymes superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase were decreased while the levels of hydrogen peroxide and lipid peroxidation were increased in the epididymal sperm as well as in the caput, corpus, and cauda epididymis after 4 or 7 days of treatment. The activities of superoxide dismutase decreased while the levels of lipid peroxidation increased in the liver but not in the kidney in all groups. Co-administration of the antioxidant vitamin E (20 mg/kg body weight/ day) to the 200 mg/kg/d methoxychlor-treated rats for 7 days prevented significant changes in the antioxidant systems in the epididymis and epididymal sperm and prevented alterations in sperm counts and motility. The results indicated that methoxychlor induces oxidative stress in the epididymis and epididymal sperm by decreasing antioxidant enzymes, possibly by inducing reactive oxygen species. In conclusion the adverse effect of methoxychlor on the male reproduction could be due to induction of oxidative stress.     

Reproductive performance, blood testosterone, lipid peroxidation and seminal plasma biochemistry of rabbits as affected by feeding Acacia saligna under subtropical conditions.

Thirty-two New Zealand White growing rabbits (eight-week old) were used to determine the effect of feeding acacia-based diets on semen characteristics, plasma testosterone, free radicals, seminal plasma enzymes and lipids. Rabbits were randomly assigned to four equal groups. The first group (control) was fed a basal diet only, and the other three groups were fed other three diets, as follows: 80% of basal diet+20% of acacia leaves (low), 60% of basal diet+40% of acacia leaves (medium) and 40% of basal diet+60% of acacia leaves (high), respectively for 32 week. Semen samples were collected throughout the last 12 week of the experimental period. Rabbits fed on different levels of Acacia showed no significant changes in libido (reaction time), ejaculate volume, sperm concentration, packed sperm volume and initial hydrogen ion concentration compared to control. However, low and/or medium levels of Acacia caused significant (P<0.05) increase in total sperm output (TSO), sperm motility (%), total motile sperm per ejaculate (TMS), normal sperm, total functional sperm fraction (TFSF), semen initial fructose, live sperm and plasma testosterone. On the other hand, high level of Acacia did not show any significant change in TSO, sperm motility (%), TMS, initial fructose, TFSF or testosterone, while live and normal sperm decreased. All levels of Acacia caused a significant decrease in the concentrations of thiobarbituric acid-reactive substances (TBARS) and an increase in the activity of glutathione S-transferease. The activities of lactate dehydrogenase (LDH), aminotransferases and phosphatases were significantly increased in seminal plasma of animals fed low or medium levels of Acacia. Seminal plasma total lipid, triglyceride, low density lipoprotein and free fatty acids were significantly (P<0.05) decreased in low or medium levels of Acacia. On the other hand, total cholesterol, percentage cholesterol (out of total lipids) and high density lipoprotein were significantly (P<0.05) increased in seminal plasma of rabbits fed on low or medium levels of Acacia. High level of Acacia did not cause any changes in the previous parameters. It can be concluded that up to 40% Acacia leaves could be used successfully and safely in the diet of rabbits without adversely affecting their reproductive performance under subtropical conditions.     

PROTECTIVE EFFECTS OF MANGANESE AGAINST LIPID PEROXIDATION

The aim of this study was to investigate the effects of chronic, daily, 30-d administration of manganese chloride (MnCl₂) to male Sprague-Dawley rats on lipid peroxidation in various tissues. Rats were intraperitoneally injected with MnCl₂ (20 mg/kg) once daily for 30 con secutive days. The Mn accumulated in liver, spleen, adrenal glands, heart, kidneys, lung, and testes. This was associated with decreased lipid peroxidation in liver, spleen, and adrenal glands and a decrease in the levels of Fe in these tissues. In a second group of animals, Mn (20 mg/kg/d) and glutathione (GSH, 15 mg/kg/d) were administered ip for 30 d. GSH counteracted the Mn-induced protective fall in lipid peroxidation, but Fe levels remained lower in liver and spleen. Mn decreases lipid peroxidation in certain tissues, which may involve lowering Fe content, but interaction with Fe is not the sole mechanism.     

Assessment of the reproductive toxicity of inhalation exposure to ethyl tertiary butyl ether in male mice with normal,low active and inactive ALDH2

No data are available regarding aldehyde dehydrogenase 2 (ALDH2) polymorphisms related to the reproductive toxicity possibly caused by ethyl tertiary butyl ether (ETBE). In this study, two inhalation experiments were performed in Aldh2 knockout (KO), heterogeneous (HT) and wild type (WT) C57BL/6 male mice exposed to ETBE, and the data about general toxicity, testicular histopathology, sperm head numbers, sperm motility and sperm DNA damage were collected. The results showed that the 13-week exposure to 0, 500, 1,750 and 5,000 ppm ETBE significantly decreased sperm motility and increased levels of sperm DNA strand breaks and 8-hydroxy-deoxyguanosine in both WT and KO mice, the effects were found in 1,750 and 5,000 ppm groups of WT mice, and all of the three exposed groups of KO mice compared to the corresponding control; furthermore, ETBE also caused decrease in the relative weights of testes and epididymides, the slight atrophy of seminiferous tubules of testis and reduction in sperm numbers of KO mice exposed to ≥500 ppm. In the experiment of exposure to lower concentrations of ETBE (0, 50, 200 and 500 ppm) for 9 weeks, the remarkable effects of ETBE on sperm head numbers, sperm motility and sperm DNA damage were further observed in KO and HT mice exposed to 200 ppm ETBE, but not in WT mice. Our findings suggested that only exposure to high concentrations of ETBE might result in reproductive toxicity in mice with normal active ALDH2, while low active and inactive ALDH2 enzyme significantly enhanced the ETBE-induced reproductive toxicity in mice, even exposed to low concentrations of ETBE, mainly due to the accumulation of acetaldehyde as a primary metabolite of ETBE.     

Protective effects of manganese against lipid peroxidation

The aim of this study was to investigate the effects of chronic, daily, 30-d administration of manganese chloride (MnCl2) to male Sprague-Dawley rats on lipid peroxidation in various tissues. Rats were intraperitoneally injected with MnCl2 (20 mg/kg) once daily for 30 consecutive days. The Mn accumulated in liver, spleen, adrenal glands, heart, kidneys, lung, and testes. This was associated with decreased lipid peroxidation in liver, spleen, and adrenal glands and a decrease in the levels of Fe in these tissues. In a second group of animals, Mn (20 mg/kg/d) and glutathione (GSH, 15 mg/kg/d) were administered ip for 30 d. GSH counteracted the Mn-induced protective fall in lipid peroxidation, but Fe levels remained lower in liver and spleen. Mn decreases lipid peroxidation in certain tissues, which may involve lowering Fe content, but interaction with Fe is not the sole mechanism.     

Influence of dietary manganese and vitamin E on adriamycin toxicity in mice

Adriamycin (ADR) administration can result in cardiac toxicity. One suggested mechanism of damage is through increased lipid peroxidation. We have evaluated the biochemical response of mice to ADR treatment when fed Mn-sufficient, vitamin E-sufficient (+Mn, +E); Mn-sufficient, low vitamin-E (+Mn, -E); Mn-deficient, vitamin E-sufficient (-Mn, +E); or Mn-deficient, low vitamin-E (-Mn, -E) diets. Mice were injected i.p. with ADR (10 mg/kg bw) or saline (0.9% w/v). ADR injection resulted in a reduction of food intake by 2 days postinjection; by 5 days postinjection ADR-treated mice lost an average of 6.0 g. Heart Mn superoxide dismutase (SOD) activity of -Mn mice was 50% that of +Mn mice; ADR did not affect MnSOD activity. Lipid peroxidation was highest in the -Mn, -E mice, being 2-fold higher than that observed in +Mn, +E mice. ADR injection did not affect lipid peroxidation. An interaction between Fe and ADR treatment was apparent; ADR injected -Mn, -E mice had liver Fe concentrations which were twice that of the saline injected mice fed -Mn, -E diets. These data show that the antioxidant status of an animal may influence ADR toxicity.     

Low lead environmental exposure alters semen quality and sperm chromatin condensation in northern Mexico

We evaluated environmental-lead (Pb) effects on semen quality and sperm chromatin, considering Pb in seminal fluid (PbSF), spermatozoa (PbSpz), and blood (PbB) as exposure biomarkers in urban men (9.3 microg/dL PbB). Several individuals (44%) showed decreases in sperm quality; sperm concentration, motility, morphology and viability associated negatively with PbSpz, whereas semen volume associated negatively with PbSF. Multiple linear regression estimated PbSF and PbSpz thresholds for alterations in semen quality. Forty-eight percent of samples showed high values of nuclear chromatin condensation (NCD) positively associated with PbSF and zinc in spermatozoa (ZnSpz). ZnSpz values were higher than in fertile men. These results suggest that Pb may affect sperm chromatin by altering sperm Zn availability. PbB was not associated with semen quality or NCD, suggesting that Pb in semen compartments assesses better the amount of Pb in the reproductive tract; therefore, these are better biomarkers to evaluate toxicity at low Pb-exposure levels.     

Novel membrane localized iron chelators as inhibitors of iron-dependent lipid peroxidation

Attachment of various iron chelating moieties to hydrophobic steroids greatly enhanced their abilities to inhibit iron-dependent lipid peroxidation. Using whole rat brain homogenates, lipid peroxidation initiated by the addition of 200 microM Fe2+ was assessed by the formation of thiobarbituric acid reactive products (TBAR). Under these conditions, 50% inhibitory concentrations of Fe3+ chelators such as desferrioxamine or N1,N8-bis(2,3-dihydroxybenzoyl) spermidine hydrobromide (compound II) were around 170 and 50 microM respectively. Coupling desferrioxamine or compound II to a steroid at the D ring increased their potency in lipid peroxidation assays by 5- to 10-fold. Evidence that inhibition of lipid peroxidation by the steroid-chelator adducts was due to iron chelation was suggested by the fact that methylation of the catechol oxygens of compound II, which are essential for chelation, completely eliminated activity of the steroid adduct. A series of 21-aminosteroids which complex Fe2+ iron and potently inhibit iron-dependent lipid peroxidation has also been synthesized. Coupling Fe2+ chelators to hydrophobic steroids increased their inhibitory potencies by as much as 10- to 100-fold. Some steroid-based Fe2+ chelators stimulated lipid peroxidation at low concentrations in the presence of Fe3+. The degree of stimulation was related to the affinity of a compound for Fe2+ with the stronger chelators causing greater stimulation. The most potent inhibitors of lipid peroxidation in the 21-aminosteroid series were found to be those compounds forming the weakest Fe2+ complexes. The findings suggest that it is iron at or near the membrane that is responsible for the catalysis of lipid peroxidation. The compounds described should provide useful tools for studies of the involvement of iron in the lipid peroxidation process.     

Comparison of the effects of metals on cellular injury and lipid peroxidation in isolated rat hepatocytes

Various mechanisms, including increases in lipid peroxidation, have been proposed to account for metal-induced cellular injury. By comparing several metals in the same cell population, it is possible to determine whether a correlation exists between ability to produce cell injury and ability to alter parameters pertaining to a particular mechanism. Of particular interest in this study was the relation between metal-induced cytotoxicity and increases in lipid peroxidation. The effects of Cr, Mn, Zn, Ni, Pb, Se, V, Fe, Cd, Hg, Cu, at final concentrations of 1-1000 microM, on the viability of isolated hepatocytes were therefore examined by assessing the loss of intracellular K+ and aspartate aminotransferase (AST). Simultaneously, the ability of the metals to induce lipid peroxidation, as measured by an increase in thiobarbituric acid (TBA) reactants, was assessed. Hg and Cu required the lowest concentration to produce cellular injury, while Cd produced less dramatic changes in cell viability and Fe at 1000 microM produced only a small decrease in intracellular K+. The largest absolute increases in lipid peroxidation were found in the presence of V, followed by Fe and Hg, with Cd and Se causing the smallest increase in TBA reactants. These observations suggest that the lipid peroxidation associated with Cd and Hg is not necessarily responsible for the loss of cell viability induced by these two metals.     

Effect of exogenous selenium on the testicular toxicity induced by ethanol in rats

The effects of supplementation of selenium at a dose of 10 microg/ kg body weight were investigated on ethanol induced testicular toxicity in rats. In the present study, four groups of male albino rats were maintained for 60 days, as follows: (1) Control group (normal diet) (2) Ethanol group (4g/kg body weight) (3) Selenium (10 microg/kg body weight) (4) Ethanol + Selenium (4g/kg body weight + 10 microg/kg body weight). Results revealed that ethanol intake caused drastic changes in the sperm count, sperm motility and sperm morphology. It also reduced the levels of testosterone and fructose. The activities of 3betaHSD, 17betaHSD in the testis and SDH in the seminal plasma were also reduced. Lipid peroxidation was also enhanced as the lipid peroxidation products were increased and the activities of the scavenging enzymes were reduced. But on coadministration of selenium along with alcohol all the biochemical parameters were altered to near normal levels indicating a protective effect of selenium. These results were reinforced by the histopathological studies.     

Effects of various compounds on lipid peroxidation mediated by detergent-solubilized rat liver NADPH-cytochrome C reductase.

A reconstituted lipid peroxidation system containing NADPH-cytochrome c reductase isolated from detergent-solubilized rat liver microsomes was used to determine the effects of several compounds, including drugs, on the lipid peroxidation activity. EDTA and ferrous ion were essential requirements for reconstitution of the activity. The addition of 1,10-phenanthroline to the system containing both EDTA and ferrous ion further enhanced the activity. Pyrocatecol, thymol, p-aminophenol, imipramine, p-chloromercuribenzoate (PCMB) and alpha-tocopherol exhibited strong inhibition, aniline, N-monomethylaniline, aminopyrine, benzphetamine, SKF 525-A and NADP exhibited moderate inhibition, and phenol, benzoic acid, acetanilide and nicotinamide exhibited less or no inhibition at the concentrations lower than 1000 micron M. Metal ions such as Hg+, Hg2+, Co2+, Cu2+, Mn2+ and U6+ inhibited lipid peroxidation strongly. In addition, Cd2+, St2+ and Ca2+ exhibited less potent to moderate inhibition, and Ba2+ and Mg2+ were without effects on the activity. Among sulfhydryl compounds tested, dithiothreitol inhibited lipid peroxidation to a greater extent than did the other three compounds, glutathione, cysteine and mercaptoethanol.     

Effects of certain mono-, di- and trivalent metal cations on the K+-stimulated p-Nitrophenyl phosphatase in rat brain

The effects of certain monovalent (Ag+1 and Li+1), divalent (Hg+2, Cu+2, Zn+2, Co+2, Fe+2, Pb+2, Mn+2, Sn+2, Ni+2, and Se+2) and trivalent (Fe+3, As+3, and Al+3) metals on a mitochondrial preparation of K+-stimulated-p-nitrophenyl phosphatase (K+-PNPPase) from rat brain were studied. Except for salts of Ni+2, Se+2 and Li+1, which irrespective of concentration failed to produce 50% inhibition, all of the metals examined were found to be potent inhibitors of the enzyme with 150 values of 0.24 microM for Ag+1 among the monovalent, 0.70, 30, 37, 38, 47, 60, 62, 490 and 850 microM for Hg+2, Cu+2, Cd+2, Zn+2, Co+2, Fe+2, Pb+2, Mn+2 and Sn+2, respectively, among the divalents and 100, 550 and 870 microM for Fe+3, As+3, and AL+3 respectively, among the trivalents. Salts of silver and mercury were the most toxic for this enzyme. All metals showed concentration dependent inhibition except lithium. The order of their potency was Ag+1 greater than Hg+2 greater than Cu+2 greater than Cd+2 greater than Zn+2 greater than Co+2 greater than Fe+2 greater than Pb+2 greater than Fe+3 greater than Mn+2 greater than As+3 greater than Sn+2 greater than Al+3 greater than Ni+2 greater than Se+2 greater than Li+1.     

Induction of oxidative stress by bisphenol A in the epididymal sperm of rats

Bisphenol A has been shown to affect the reproduction of male rats and mice. However, the mechanism of action of bisphenol A on the epididymal sperm is not elucidated. The present study was undertaken to evaluate the effect of bisphenol A on the antioxidant system of rat epididymal sperm. Bisphenol A was administered orally to male rats at the dose levels of 0.2, 2 and 20 microg/Kg body weight per day for 45 days. After 24 h of the last treatment, rats were weighed and killed using anesthetic ether. The body weight of treated rats did not show significant change as compared with the corresponding control groups. In bisphenol A-treated rats there was a significant decrease in the weight of the testis and epididymis; the weight of ventral prostate increased significantly whereas there was no significant change in the weight of seminal vesicles as compared with the corresponding group of control animals. Sperm collected from the epididymis were used for sperm count and biochemical estimations. Administration of bisphenol A caused a reduction in the epididymal sperm motility and sperm count in a dose-dependent manner. The activities of superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase were decreased while the levels of H(2)O(2) and lipid peroxidation increased significantly in the treated rats as compared with the corresponding group of control animals. The results suggested that graded doses of bisphenol A elicit depletion of antioxidant defence system and induce oxidative stress in epididymal sperm of rats. In conclusion, the adverse effect of bisphenol A on male reproduction may be due to induction of oxidative stress in sperm.     

Reproductive toxicity of 1-bromopropane, a newly introduced alternative to ozone layer depleting solvents, in male rats.

1-Bromopropane has been newly introduced as an alternative to ozone-depleting solvents. We aimed to clarify its dose-dependent reproductive toxicity in male rats. Thirty-six Wistar male rats were randomly divided into 4 groups of 9. The groups were exposed to 200, 400, or 800 ppm 1-bromopropane or only fresh air, 8 h per day for 12 weeks. Epididymal sperm indices were evaluated after a 12-week exposure. The testes, epididymides, seminal vesicle, prostate, and other organs were weighed and examined histopathologically. Spermatogenic cells, in stage VII seminiferous tubules, and retained spermatids, at the basal region of stages IX-XI seminiferous epithelium, were counted. Plasma testosterone levels were measured by radioimmunoassay. The testicular weight did not significantly change, but the weight of epididymides, seminal vesicle, and prostate dose-dependently decreased. The weight of seminal vesicle decreased significantly at the lowest concentration of 200-ppm and over. 1-Bromopropane induced a dose-dependent decrease in the epididymal sperm count and in motility, as well as an increase in tailless sperm and sperm with an immature head shape. The spermatogonia, preleptotene spermatocytes, pachytene spermatocytes, and round spermatids did not decrease significantly at stage VII. Retained, elongated spermatids near the basement membrane at the postspermiation stages IX-XI increased dose-dependently. Plasma testosterone levels significantly decreased at the 800-ppm dosage. 1-Bromopropane caused failure of spermiation. Its reproductive toxicity is different from that of 2-bromopropane, which specifically impairs spermatogonia. Thus, this solvent may have serious reproductive toxic effects in men, and should be used very cautiously in the workplace.     

Comparison of the effects of Fe2+ and Cu2+ on prostaglandin synthesis in rabbit kidney medulla slices

The effects of Fe2+ and Cu2+ on the generation of medullary prostaglandins E2 and F2 alpha have been compared. Fe2+ markedly promoted the lipid peroxidation of rabbit kidney medulla slices. The lipid peroxidation induced by Fe2+ inhibited both prostaglandin E2 and prostaglandin F2 alpha formation to a similar extent. While Cu2+ produced only a small increase in lipid peroxidation, it had a powerful inhibitory effect on prostaglandin E2 formation. Simultaneously, prostaglandin F2 alpha production was increased. In the presence of Cu2+ the net increased amount of prostaglandin F2 alpha was much smaller than the net decreased amount of prostaglandin E2 (15-20%). These results suggest that Cu2+ has the potential to modulate prostaglandins E2 and F2 alpha synthesis by affecting endoperoxide E2 isomerase or endoperoxide reductase independent of its peroxidative action.