Objective: To evaluate normal spermatozoa morphometry and ultrastructure in estuarine crocodile (Crocodylus porosus). Methods: Four adult male crocodiles aged between 12-15 years, with an average snout to tail length of (3.15±0.01) m were selected for this study. Manipulation of the phallus digitally from the base of the penis was performed on four adult male crocodiles to facilitate the flow of semen to the sulcus. Semen was collected from all individuals for spermatozoa morphometry and ultrastructure study. Morphometry analysis was performed from eosin-nigrosin stained spermatozoa sample. Scanning electron microscopy was conducted to observe the surface ultrastructure of spermatozoa. Results: The morphology of crocodile spermatozoa was made up of acrosome, head, and tail which corresponded to (5.55±1.20) μm, (12.74±1.57) μm, and (70.67±4.40) μm, respectively. The total length of spermatozoa in estuarine crocodile was measured at (88.96±0.52) μm. The most common spermatozoa abnormalities found in the samples were cytoplasmic droplets. Conclusions: Normal morphology, morphometry of spermatozoa in estuarine crocodile which are vermiform shaped-head with a long tail along with questionably high cytoplasmic droplets count are described in this study.

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Objective: To assess the effect of short-term gavage of ethanolic extract of Imperata cylindrica L root on the ovarian activity and estrus behavior of female mice. Methods: Eighteen virgin female ddY mice, 8 to 10 weeks of age, weighing 22–25 g with regular estrus cycle, were divided into three groups. Group 1 received 0.5% carboxymethylcellulose, whereas Groups 2 and 3 received the ethanolic extract of Imperata cylindrica L root at 90 and 115 mg/kg body weight (b.w.) per day by gavage for 20 days, respectively. All of the groups were checked before 9 a.m. daily for vaginal cytology to determine the estrus phase. On day 21, the mice were sacrificed to collect serum samples to quantify the concentrations of reproductive hormones using enzyme-linked immunosorbent assay and to determine changes in the reproductive organs based on their reproductive organ weight, histomorphology, and histomorphometry of ovarium and uterus. Results: The reproductive organ weight in the treatment groups was similar compared with that in the control group. The 90 mg/kg b.w. treatment group showed an increase in corpus luteum number when compared with the control group, with few degenerated follicles and diminished oocytes. Moreover, the 115 mg/kg b.w. treatment group showed fewer primordial and primary follicles and an increase in corpus luteum number and a prolonged diestrus phase compared to the control and 90 mg/kg b.w. treatment groups. The histomorphology examination of the uterus showed that the thickness of myometrium and epithelium in the treated animals was similar to the control group. In addition, there was a significant decrease in follicle-stimulating hormone level in the 115 mg/kg b.w. treatment group (P<0.05). Conclusions: Short-term gavage of ethanolic extract of Imperata cylindrica L root reduces the follicle-stimulating hormone serum level and folliculogenesis.

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Objective: To investigate the protective effects of Mucuna (M.) pruriens against arsenic-induced testicular impairment in albino mice. Methods: Thirty-six male albino mice were divided into six groups, with 6 mice in each group. Group 1 received drinking water as the normal control. Groups 2 to 6 received sodium arsenite (40 mg/L) in drinking water. Group 3 received sodium arsenite (40 mg/L) and 300 mg/kg body weight (b.w.) of N-acetylcysteine. Group 4 received sodium arsenite (40 mg/L) and 500 mg/kg b.w. of M. pruriens. Group 5 received sodium arsenite (40 mg/L) and 1 000 mg/kg b.w. of M. pruriens. Group 6 received sodium arsenite (40 mg/L) and 2 000 mg/kg b.w. of M. pruriens. N-acetylcysteine and M. pruriens were administered orally once a day. Animals were subjected to the above treatments for 45 days. Animals were sacrificed with overdose of ketamine 24 h following drug administration. The testis was used for biochemical estimations (lipid peroxidation and glutathione), and the epididymis was used to determine the sperm count and morphology. Results: Sodium arsenite significantly decreased (P <0.01) the sperm count and glutathione levels of the testis. It significantly increased (P <0.01) the abnormal architecture of the spermatozoa and lipid peroxidation levels. Treatment with M. pruriens significantly increased the sperm count and the glutathione levels [500 mg/kg b.w. and 1 000 mg/kg b.w. (P <0.01)]. A significant decrease in sperm abnormality and lipid peroxidation levels [500 mg/kg b.w. and 1 000 mg/kg b.w. (P <0.01)] was also observed. Conclusions: The perturbed sperm parameters and antioxidant levels of the arsenic insulted testis are attenuated by 500 mg/kg b.w. and 1 000 mg/kg b.w. M. pruriens.

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Objective: To investigate the hormonal, histopathological toxicity and sperm quality of ciprofloxacin in male genital system in the rabbit model. Methods: Twenty adult New Zealand male rabbits were randomly divided into the treatment and control groups, and respectively received 30 mg/kg/day ciprofloxacin and normal saline intraperitoneally for 14 days. Blood samples and testicular tissues were taken for testosterone by enzyme linked immunosorbent assay method, and measurement of histopathologic and histomorphometric investigations was made on the 14th day of the experiment and 56 days after the last dose of ciprofloxacin. Additionally, epididymis sperm sample was collected for quality analysis. Results: On day 14 of the experiment, histopathologic examination revealed severe degenerative changes in the epithelium of seminiferous tubules and loss of all germ cells in most tubes in the treatment group. In the histomorphometric study, significant reductions in the seminiferous tubules diameter, epithelium height, and Johnsen’ score were observed. On day 56 after the last dose of ciprofloxacin, the seminiferous tubules revealed regeneration in the treatment group and no significant difference was observed in the spermatogenesis parameters except epithelial height parameter between the control group and the treatment group. All sperm quality parameters were significantly decreased on day 56 after the last dose of ciprofloxacin. Testosterone levels did not significantly change during the follow-ups period. Conclusions: There is potential reversible testicular toxicity for ciprofloxacin in the rabbit model in according to relative regeneration of spermatogenic epithelium after 56 days of last dose of ciprofloxacin. Sperm quality will be improved with a delay after complete regeneration of seminiferous tubules.

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Objective: To study the influence of N-acetylcysteine on the pituitary-gonadal axis hormones and protamine expression level in streptozotocin-induced diabetic male rats. Methods: Forty-two adult male Wistar rats were divided into 6 groups, with 7 rats in each group. The control group left untreated; the streptozotocin group only received 50 mg/kg body weight streptozotocin intraperitoneally for 5 days to induce diabetes; the N-acetylcysteine group only received 200 mg/kg body weight N-acetylcysteine intraperitoneally, and the streptozotocin+N- acetylcysteine groups 1, 2 and 3 received 50 mg/kg streptozotocin intraperitoneally for 5 days to induce diabetes and then received 100, 200 and 400 mg/kg body weight doses of N-acetylcysteine intraperitoneally for 28 days, respectively. Enzyme-linked immunosorbent assay was used to measure the serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone, and real-time PCR was applied for measuring protamine expression level. Results: Compared to the control and N-acetylcysteine groups, a significant decrease in the body weight, testicular weight and levels of testosterone and protamine expression was observed in the streptozotocin group and the streptozotocin+N-acetylcysteine groups 1 and 2. On the contrary, the levels of LH and FSH increased significantly. In the streptozotocin+N-acetylcysteine group 3, the body weight, testicular weight and expression level of protamine were significantly higher than those of the streptozotocin group. In the streptozotocin+N-acetylcysteine groups, testosterone and LH levels were significantly higher than and lower than the streptozotocin group, respectively. In the streptozotocin+N- acetylcysteine groups 2 and 3, the level of FSH was significantly lower than that of the streptozotocin group and streptozotocin+N- acetylcysteine group 1. Furthermore, a significant increase in the expression level of protamine was observed in the streptozotocin+N- acetylcysteine groups 2 and 3 when compared to the streptozotocin group and streptozotocin+N-acetylcysteine group 1. Conclusions: N-acetylcysteine in an optimal dose of 400 mg/kg body weight has a protective influence on the pituitary-gonadal axis hormones and also on the expression level of protamine in diabetic male rats.

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Objective: To explore the association between statin therapy and the risk of erectile dysfunction by literature review. Methods: We conducted diversities of search strategies including electronic database searches of MEDLINE, Scopus, Pubmed and Web of Science using MeSH terms, keywords and title words during the search. Reference lists of identified and public articles were reviewed. In addition, only English articles were considered and case reports were not concerned in the review. The key features of recognized applicable search studies were considered and the conclusions were summarized in a narrative review. Results: Different studies gave a consensus that erectile dysfunction was regarded as an early sign of silent cardiovascular disorder and hidden atherosclerosis. Different studies reported that statins might induce erectile dysfunction through induction of peripheral neuropathy, cognitive deficits, and reduction of circulating testosterone. However, most of recent studies illustrated that statins led to a significant improvement in erectile function and sexual health in men with age over forty years. Atorvastatin advanced endothelial nitric oxide concentrations through activation and upregulation of endothelial nitric oxide synthase and rescued phosphodiesterase-5 inhibitors non-responders since nitric oxide and cyclic guanosine monophosphate increased penile blood flow and improved erectile function. Conclusions: According to the assorted view of preponderance, statins improved erectile dysfunction is more dominant than statins induced erectile dysfunction. Therefore, statins regardless of its property improve erectile dysfunction through amelioration of penile endothelial dysfunction, and penile neuronal reflexes that are inter-related during sexual excitation and penile erection.

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Objective: To determine the effects of routine iron supplementation on oxidative stress markers in pregnant women. Methods: This study enrolled 122 pregnant women aged between 20-38 years throughout three stages of pregnancy: beginning with pregnancy, until the twentieth week and up to the thirtieth week. They were divided into two groups: 61 pregnant women not supplemented with iron (the control group), and 61 pregnant women supplemented with iron (the iron group). Blood samples were collected and protein, iron, uric acid, total bilirubin level and oxidative stress status were estimated using molecular spectrophotometric method. Copper levels were estimated by using atomic absorption spectroscopy method. Results: Iron, protein and copper concentrations were significantly increased (P<0.05) at 20 and 30 weeks of pregnancy and hemoglobin level was significantly increased (P<0.05) at the beginning of pregnancy and until 20 weeks in the iron group compared to the control group. In addition, levels of serum uric acid, serum total bilirubin and malondialdehyde in serum and erythrocytes were significantly increased at 20 and 30 weeks of pregnancy whereas the reduced glutathione level was significantly decreased in serum (30 weeks of pregnancy) and erythrocytes (20 and 30 weeks of pregnancy) in the iron group as compared to the control group. Serum oxygen radical absorbance capacity was not significantly changed. There was a significant positive correlation between serum iron level and serum copper, malondialdehyde level in serum and erythrocytes in pregnant women supplemented with iron. Conclusions: Iron supplement at 20 and 30 weeks of pregnancy plays a role in the development of oxidative stress which is the origin of several maternal-fetal complications.

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Objective: To evaluate the cryoprotective effect of butylated hydroxytoluene on buck frozen semen. Methods: Semen was collected from Boer (n=6) and Zaraibi (n=6) bucks by electroejaculator for 5 weeks. Semen aliquots were diluted at 38 °C in Tris-buffer with egg yolk 15.0% (vol/vol) (Tris- egg yolk extender) or soya lecithin 2.5% (weight/vol) (Tris-soya lecithin extender) supplemented with butylated hydroxytoluene at 0.0 (as the control), 0.5, 1.0, 2.0 and 4.0 mM. Post-thawing motility (at 400x magnification), plasma (hypo-osmotic swelling test), acrosome (Trypan blue/Giemsa dual staining) membranes, DNA (comet assay), and lipid peroxidation (by malondialdehyde concentration) were assessed. Results: Spermatozoa motility was enhanced by butylated hydroxytoluene in Tris-soya lecithin extender at 0.5 mM in the two breeds, and in Tris-egg yolk extender at 1.0 mM in Boer and at 2.0 mM in Zaraibi bucks for up to 3 h post-thawing. Plasma and acrosome membranes and DNA integrity of the two breeds were maximally high with butylated hydroxytoluene at 1.0-2.0 mM in Tris-egg yolk extender and at 0.5-1.0 mM in Tris-soya lecithin extender. Lipid peroxidation was minimal with butylated hydroxytoluene at 1.0-2.0 mM in Tris-egg yolk and soya lecithin extenders in the two breeds. Butylated hydroxytoluene at 4.0 mM deteriorated spermatozoa motility, and plasma and acrosome membranes. Conclusions: The consequence of butylated hydroxytoluene on buck frozen-thawed spermatozoa varies with the levels of supplementation, buck breed, and phospholipid source in the extender. Semen parameters of Boer buck are better in their response to butylated hydroxytoluene than Zaraibi buck. Butylated hydroxytoluene at 1.0 and 2.0 mM in Tris-egg yolk extender, and at 0.5 mM in Tris-soya lecithin extender represents the best concentrations and profitably improves the semen quality of buck semen.

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