Objective: To investigate the effect of Camellia (C.) sinensis in mitigating oxidative damage and reproductive toxicity in testis induced by sodium fluoride in a rat model. Methods: Twenty-four adult male Wister rats were divided into 4 groups, with 6 rats in each group. Group 1 orally received distilled water (1 mL/100 g body weight) daily and served as the control group, while group 2 received drinking water with 100 ppm sodium fluoride per day for 21 consecutive days, group 3 was administered with only C. sinensis extract by gavage at a dose of 100 mg/kg body weight and group 4 received drinking water with 100 ppm sodium fluoride and 100 mg/kg body weight C. sinensis leaf extract per day for 21 consecutive days. At the end of the treatment, the rats were sacrificed under light ether anesthesia. The gonado-somatic index, sperm count and motility, serum level of luteinizing hormone and testosterone were assayed. Lipid peroxidation [malondialdehyde (MDA) level], nitric oxide (NO) production, and activities of antioxidant enzymes - superoxide dismutase (SOD), catalase, and reduced glutathione level (GSH) were also analysed. Results: Sodium fluoride treatment significantly decreased gonado-somatic index, sperm count and motility as well as the serum level of luteinizing hormone and testosterone (P<0.05). The histological examination of testes revealed atrophy and degenerative changes in several seminiferous tubules, along with enhanced interstitial space and a reduced number of Leydig cells. There was a highly significant increase in NO and MDA production (P<0.05), while SOD, catalase activities and GSH level decreased significantly (P<0.05). However, C. sinensis significantly restored testicular weight, sperm parameter, hormonal level (P<0.05), and also reversed MDA and NO generation and antioxidant enzymes activities in the testicular tissue (P<0.05). Conclusions: C. sinensis may have an ameliorative role against sodium fluoride-induced oxidative damage in the testis probably because of its antioxidant property.

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Early diagnosis of pregnancy plays an important role to minimize reproductive losses in farm animals. There are several methods for pregnancy diagnosis like profiling of reproductive hormones (such as progesterone and estrone sulfate), but sometimes they provide false-positive results. Embryo specific pregnancy markers, which delineate the presence and viability of the embryo, are considered as perfect for pregnancy determination. Pregnancy-associated glycoproteins are distinguished as the best indicator for the determination of early pregnancy, fetal number, and birth weight of kids. Pregnancy-associated glycoproteins are structurally correlated to aspartic proteinase and are communicated in the external epithelial cell layer of the placenta. They have been found to share about half amino acid sequence identity with pepsinogen, pepsin, cathepsin D and E. Dislike different individuals from aspartic proteinase family, numerous pregnancy-associated glycoproteins appear to be latent compound as a result of amino acid substitutions in and around the catalytic site. This review is to discuss the scope and prospects of pregnancy-associated glycoproteins as a pregnancy marker in farm animals, more specifically in goats.

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Objective: To investigate morphokinetic characteristics of embryos displaying either reverse cleavage or direct cleavage during the first, second or third cleavage cycle. Methods: A total of 167 in vitro fertilization and/or intracytoplasmic sperm injection treatment cycles undertaken by 167 women [aged (35.0±4.6) years] were included for reverse cleavage analysis, and a total of 167 in vitro fertilization and/or intracytoplasmic sperm injection treatment cycles undertaken by 167 women [aged (33.8 ±4.3) years] were included for direct cleavage analysis in this study. Using a sibling-embryo design, morphokinetic profiles (both before and after the onset of abnormal event) of embryos displaying reverse cleavage (n=241) or direct cleavage (n=244) were compared with their unaffected siblings (the controls) in the first, second and third cell cycles (n=32, n=142, n=562; n=195, n=412, n=205, respectively), at different developmental stages up to day 3. Results: Direct cleavage embryos demonstrated significantly delayed cleavage rates prior to the event regardless of developmental stage of the occurrence, while reverse cleavage embryos showed similar cleavage rates to their unaffected siblings. Post event, direct cleavage embryos sped up cleavage rates while reverse cleavage embryos slowed down. Conclusions: Altered morphokinetic profiles are displayed by direct cleavage embryos both before and after their occurrence and reverse cleavage embryos after the occurrence, which could potentially confound morphokinetic comparisons if not separated from their unaffected sibling embryos. Further study is warranted in order to fully understand the biological mechanisms of such events.

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Objective: To identify the alteration of tyrosine phosphorylated protein expression in rats with polycystic ovary syndrome (PCOS). Methods: Sixteen female Sprague-Dawley rats were divided into the control and letrozole-induced PCOS groups. The oestrus cycle of rats was performed by vaginal smear. Sex hormones and morphology of the ovary, oviduct, and uterus were observed. Expressions and intensity of androgen receptor and tyrosine phosphorylated proteins of reproductive organs were investigated by Western blot. Results: Various polycysts and increased androgen receptor expression were present in the ovary of the PCOS group. The levels of follicle-stimulating hormone and testosteone were significantly higher in the PCOS group while progesterone and estradiol levels were significantly decreased as compared with the control group (P<0.05). Only the size of uterus in the PCOS group was significantly smaller than the control group. However, the density of collagen fibers observed in PCOS uterus was greater than the control group. Moreover, tyrosine phosphorylated proteins were significantly overexpressed in ovary (52, 42, and 28 kDa), oviduct (72, 56, 42, and 28 kDa), and uterus (53 and 42 kDa) of the PCOS group compared to the control group. Conclusions: Presence of tyrosine phosphorylated proteins in the ovary, oviduct and uterus suggests that overexpression of tyrosine phosphorylated proteins may be involved in potential mechanism of female infertility especially in PCOS.

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Objective: To investigate the hepatic and reproductive toxicity of dichlorvos and lead acetate on male Wistar rats. Methods: Fifteen adult male Wistar rats (170-190 g) were randomly divided into three groups, with 5 rats in each group. Group 1 received 0.5 mL distilled water orally and served as the control group, while groups 2 and 3 were orally treated with 2 mg/kg body weight (b.w.) dichlorvos and 10 mg/kg b.w. lead acetate, respectively, for 55 days. Epididymal sperm, serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone concentrations, testicular 17 β-hydroxysteroid dehydrogenase activity (17β-HSD), androgen receptor expression, aspartate aminotransferase (AST), alanine aminotransferase (ALT), testicular oxidant and antioxidant enzymes were evaluated with standard methods. Results: Sperm count, motility, morphology, FSH, LH, testosterone levels, 17β-HSD, androgen receptor expression, and catalase activity were significantly reduced in the dichlorvos and lead acetate treated groups as compared with the control group (P<0.05). The liver AST, ALT activities and malondialdehyde concentration were significantly increased in the dichlorvos and lead acetate treated groups as compared with the control group (P<0.05). Conclusions: The reproductive and hepatic toxicity activities of dichlorvos and lead acetate in male Wistar rats are similar.

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Objective: To evaluate the development of ovarian follicles in female albino rats following vitamin D3 supplementation. Methods: Eighteen prepubertal female albino rats, aged 3-4 weeks, weighing (70.25±9.16) g, were assigned to three groups (n=6 in each group). Group A was treated with 5.00 mL/kg of distilled water and served as the control group, group B was treated with 0.025 mg/kg of vitamin D3 dissolved in distilled water, and group C was treated with 0.125 mg/kg of vitamin D3 dissolved in distilled water. All treatments were administered orally, twice weekly for 28 days. Blood and ovaries were harvested under anaesthesia. Serum vitamin D3 levels were determined by using spectrophotometric method. Ovaries were processed for histology and every10th hematoxylin and eosin stained-section was selected for histomorphometry. The number of follicles at each developmental stage was estimated. Results: Both 0.025 mg/kg and 0.125 mg/kg of vitamin D3 significantly increased serum concentrations of vitamin D3 and calcium (P<0.05), but did not alter inorganic phosphorus concentration (P>0.05). The control group had fewer growing follicles (primary, secondary and antral follicles) and more non-growing follicles (primordial and atretic follicles) when compared with the vitamin D3-supplemented groups (P<0.05). Vitamin D3 at 0.025 mg/kg significantly increased antral follicles and corpora lutea counts (P<0.05). Vitamin D3 at 0.125 mg/kg significantly increased total, primordial and atretic follicles counts (P<0.05), but significantly decreased primary, secondary, antral follicles count, ovarian weight, relative ovarian weight, and ovarian surface area when compared with the control group and rats treated with 0.025 mg/kg of vitamin D3 (P<0.05). Conclusions: Vitamin D3 supplementation at 0.025 mg/kg can enhance optimal ovarian follicle recruitment and development in female rats.

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Objective: To investigate the effects of soft agar on in-vitro proliferation of neonate mouse spermatogonial stem cells co-cultured with Sertoli cells. Methods: Tissues of neonate NMRI male mice testes were used for harvesting spermatogonial stem cells and Sertoli cells. After cell harvest, flow cytometry using promyelocytic leukemia zinc-finger (PLZF) protein antibody was used to assess the purity of the cells. The isolated testicular cells were cultured in the absence (the control group) or presence of soft agar-coated dishes (the experimental group) supplemented with leukemia inhibitory factor and glia cell line–derived neurotrophic factor for two weeks. Alkaline phosphatase activity was assessed in the colonies formed after two weeks of culture by alkaline phosphatase staining. On day 14 of culture, the expression levels of DNA-binding protein inhibitor (ID-4) and PLZF genes in the undifferentiated cells were evaluated by the detection of PLZF protein antibody using real-time PCR and immunocytochemistry techniques. The number and diameter of the colonies of spermatogonial stem cells were assessed by ImageJ software. Results: In the experimental group, the number and the diameter of colonies significantly increased as compared with those in the control group (P<0.05, P<0.01, respectively). In addition, the level of expression of ID-4 and PLZF genes in the undifferentiated cells significantly increased in the experimental group as compared with the control group (P<0.01). However, the expression of tyrosine-protein kinase kit (c-kit) gene in differentiated cells decreased in the experimental group as compared with the control group, but there was no significant difference between the two groups. Conclusions: Spermatogonial stem cells can be efficiently proliferated by culturing the stem cells in soft agar-coated dishes. The new protocol used in this study can be a valuable method for future studies.

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