Objective: To analyze the relationship between body mass index (BMI) before pregnancy and gestational weight gain throughout pregnancy with the incidence of preeclampsia. Methods: This was a systematic review-meta analysis of literature collected from three e-databases: Scopus, PubMed, and Science Direct. Quality assessment was measured with the Effective Public Health Practice Project methods. Meta-analysis was done by calculating the fixed and random-effects of odds ratio (OR) for each BMI category and gestational weight gain as compared with the incidence of preeclampsia. Results: Overweight was associated with a significantly increased risk of preeclampsia (OR=2.152, 95% CI 1.363-3.400; P=0.001). Obesity was also associated with a noticeably increased risk of preeclampsia (OR=2.856, 95% CI 1.755-4.649; P<0.001). Meanwhile, underweight was associated with a significantly reduced risk of preeclampsia (OR=0.639, 95% CI 0.500-0.817; P<0.001) when compared with normal BMI. Pregnant women who gained weight below the standard throughout pregnancy was a protective factor from preeclampsia (OR=0.813, 95% CI 0.610-1.083; P=0.157) whereas pregnant women who gained weight above the standard had almost doubled risk of preeclampsia (OR=1.850, 95% CI 1.377-2.485; P<0.001). Conclusions: The result of this study affirms the role of overweight-obesity pre-pregnancy, and gestational weight gain above the standard during pregnancy as significant risk factors for developing preeclampsia.

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Objective: To assess the therapeutic potential of ethanolic extract of Azadirachta (A.) indica in rats with polycystic ovary syndrome (PCOS). Methods: Thirty-five prepubertal female Sprague Dawley rats were randomly divided into five groups with 7 animals in each group. Group 1 received 0.5% carboxy methyl cellulose orally. Groups 2 to 5 received testosterone propionate (0.2 mg/kg, s.c.) dissolved in olive oil daily for 42 days to induce PCOS. In addition, group 3 was administered with A. indica extract (100 mg/kg, 0.5% carboxy methyl cellulose orally) from the 7th to 12th week, group 4 received quercetin (100 mg/kg, 0.5% carboxy methyl cellulose orally) and group 5 received wartmannin (100 mg/kg, 0.5% carboxy methyl cellulose orally). At the end of treatment, blood was collected for biochemical evaluation. Total follicular count and uterus corpus luteum count followed by PI3K gene expression in the ovary and uterus were evaluated. Results: The ethanolic extracts of A. indica significantly reduced body weight, ovary weight and uterus weight of rats. Extracts of A. indica also significantly increased the levels of serum glucose, total cholesterol, triglyceride, low-density lipoprotein, very low-density lipoprotein, insulin, testosterone, and luteinizing hormone. Treatment also reduced lipid peroxidation and increased antioxidant parameters in the liver homogenates of PCOS-induced rats. Histological examination of the ovary and uterus confirmed PCOS occurrence and remission state in the PCOS-induced and treated groups, respectively. Moreover, A. indica and quercetin significantly downregulated PI3K gene expression. Histopathological results of the ovary and uterus also proved the protective role of A. indica. Conclusions: A. indica leaf extract has beneficial effects in the treatment of PCOS by downregulation of PI3K gene expression.

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Objective: To investigate the testicular oxidative stress and apoptosis status, as well as the sperm functional parameters in streptozotocin (STZ) induced diabetic rats following treatment with rooibos (Aspalathus linearis), honeybush (Cyclopia intermedia) and sutherlandia (Lessertia frutescens) infusions. Methods: Diabetes was induced by injecting fourteen-week-old adult male Wistar rats (250-300 g) with a single intraperitoneal injection of STZ (45 mg/kg body weight). Fifty rats were randomly divided into five groups: the vehicle group received 0.1 M citrate buffer, the diabetic control group received 45 mg/kg STZ, the diabetic+rooibos group received 45 mg/kg STZ + 2.0% rooibos, the diabetic+honeybush group received 45 mg/kg STZ + 4.0% honeybush, and the diabetic+sutherlandia group received 45 mg/kg STZ + 0.2% sutherlandia. Rats were sacrificed 7 weeks after induction of diabetes mellitus. The testes and epididymides were harvested and weighed after induction. Spermatozoa were retrieved from the cauda epididymis for motility, concentration, and morphology analysis, and the testis was used for all biochemical assays. Oxidative stress was determined by measuring malondialdehyde levels, catalase, and superoxide dismutase activities, while apoptotic biomarkers were evaluated by Western blotting assays. Results: After induction of diabetes, rats in the diabetic control group, diabetic+rooibos group, diabetic+honeybush group, and diabetic+sutherlandia group presented with significantly elevated blood glucose levels as compared with the vehicle group (P<0.001). Rats in the diabetic control group had a reduction in sperm progressive motility, while rats in the diabetic+rooibos group and the diabetic+sutherlandia group displayed an increase in progressive motility as compared with the diabetic control group. The diabetic control animals showed a 40.0% decrease in sperm concentration when compared to the vehicle group, and there were no significant differences in sperm kinematic and speed parameters between the groups. In addition, the percentage of morphologically normal spermatozoa was increased by 13.0%, 16.0%, and 15.0% after treatment with rooibos, honeybush, and sutherlandia, respectively and the rats in the diabetic+infusion groups also displayed an increase in superoxide dismutase activity when compared to the diabetic control group. Conclusions: Rooibos, honeybush and sutherlandia infusions may partly alleviate diabetes-induced sperm function impairment by reducing oxidative stress.

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Objective: To investigate the effects of 30-day treatment with therapeutic dose equivalent levels of tramadol on serum testosterone level, sperm parameters, and testicular histology in rats. Methods: Thirty-five Wistar rats were equally divided into seven groups. Group 1 (the control group) received distilled water (0.5 mL) daily for 30 days. Groups 2-4 were gavaged with therapeutic dose equivalent levels of tramadol (1.25, 2.50 and 5.00 mg/kg/day body weight, respectively) in two equal divided doses for 30 consecutive days, and sacrificed on day 31. Groups 5-7 received similar tramadol treatments as above but they were allowed for another 30 days to recover after receiving the last dose and sacrificed on day 61 for reversibility study. Serum testosterone level and epididymal sperm were analyzed, and histopathological examination of the testis was carried out. Results: Tramadol treatment significantly decreased serum testosterone levels compared with the control group. Furthermore, tramadol treatment inhibited sperm motility and significantly and dose-dependently decreased sperm count and viability compared with the control group. In addition, tramadol significantly increased morphological abnormalities in sperm (P<0.05). The above effects of tramadol were reduced in the reversible groups. Testis histopathological examination revealed disintegrated cell architecture, eroded and atrophied seminiferous tubules, and a marked decrease in the number of spermatogenic cells in the tramadol treated groups. The histopathological changes were restored in the reversible groups, but improvement was not complete in the 5.00 mg/kg tramadol treated reversible group. Conclusions: Long term treatment with tramadol at clinical dose levels may adversely affect testosterone level, sperm parameters, and testicular histology, but they are reversible at lower doses.

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Objective: To study the effect of L-ascorbic acid supplementation on the in vitro development of buffalo embryos and evaluate the relative mRNA abundance of some pro-apoptotic, anti-apoptotic, and embryonic development-related genes. Methods: In experiment 1, we evaluated the effect of the addition of 0 (control), 50, and 100 μM L-ascorbic acid to the in vitro maturation medium on the developmental competence in terms of blastocyst rate and relative mRNA abundance of some pro-apoptotic (BAX, BID), anti-apoptotic (BCL-XL, MCL1), and embryonic development (GDF9, BMP15) related genes. Based on the results, we chose 50 μM as the suitable dose of L-ascorbic acid for the subsequent experiments. We further evaluated the blastocyst rates following the addition of 50 μM L-ascorbic acid to the in vitro culture medium (experiment 2), and in vitro maturation and in vitro culture media (experiment 3). In all three experiments, the maturation and culture media devoid of L-ascorbic acid served as the control group. Results: The blastocyst rate after adding 50 μM L-ascorbic acid to the in vitro maturation medium was significantly higher than the control group (P<0.05), whereas 100 μM L-ascorbic acid exhibited a negative effect on the blastocyst rate. The blastocyst rates for embryos cultured in 50 μM L-ascorbic acid in the in vitro culture medium alone and both in vitro maturation and in vitro culture media were significantly higher than their corresponding control groups (P<0.05). The relative mRNA abundance of BAX significantly decreased in blastocysts produced after the addition of 50 μM L-ascorbic acid as compared with the control group (P<0.05), whereas, for MCL1, it significantly decreased in blastocysts produced after the addition of 100 μM L-ascorbic acid (P<0.05). Conclusions: The supplementation of 50 μM L-ascorbic acid to in vitro maturation and in vitro culture media supports in vitro embryonic development in buffaloes by improving developmental competence and altering the expression of apoptosis-related genes.

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Objective: To evaluate the effect of cooling rates, selenium supplementation, and three semen extenders on cooled and frozen-thawed Saanen buck sperm. Methods: Twenty Saanen bucks were divided into two groups: the selenium supplemented group and the control group. Ejaculates were collected once weekly by artificial vagina. The first experiment examined the effects of cooling rates and selenium supplementation on semen characteristics of Saanen bucks. Pooled semen was diluted with triladyl extender and split into two aliquots for slow and fast cooling. The second experiment explored the effect of selenium supplementation and semen extenders on post-cryopreserved sperm quality. Ejaculates from each group were divided into three aliquots and diluted with three extenders (i.e. clarified egg-yolk, whole egg yolk, Tris without egg-yolk extenders). All samples were cooled for 2 h at 4 °C and frozen at -196 °C for 24 h. The sperm characteristics such as sperm motility, acrosome integrity, normal morphology, and viability were evaluated by using a phase-contrast microscope. Results: In the first experiment, all sperm characteristics were significantly increased in selenium supplemented samples when slow cooling was used (P<0.05). In the second experiment, in cooled semen, sperm motility and viability were significantly increased in both clarified egg yolk and Tris without egg yolk extenders in selenium supplemented samples as compared with whole egg yolk extender, respectively (P<0.05). After freezing-thawing, all sperm parameters of selenium supplemented samples in clarified egg yolk extender were significantly greater than those in Tris without egg yolk extender (P<0.05). However, normal morphology and acrosome integrity of selenium supplemented samples in whole egg yolk extender were similar to those of selenium supplemented samples in clarified egg yolk extender. Conclusions: The characteristics of cooled and post-cryopreserved sperm are greater when clarified egg yolk extender is used in semen from selenium supplemented bucks.

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