Effects of grape seed proanthocyanidin extract on side effects of high-dose methylprednisolone administration in male rats.

In this study, we investigated the effects of grape seed proanthocyanidin extract (GSPE) against the side effects of high-dose administration of methylprednisolone (MP) in male rats. A total of 32 adult Wistar male albino rats were divided into four groups: (1) control (CON), received standard food only; (2) MP, received standard food + intraperitoneal injection of 60 mg/kg MP on day 7; (3) GSPE, received standard food + 200 mg/kg/day GSPE; and (4) MP + GSPE, received standard food + 200 mg/kg/day of GSPE + intraperitoneal injection of 60 mg/kg MP on day 7. All animals in the GSPE and GSPE + MP groups were treated once a day by oral gavage for 14 consecutive days. The feed intake of rats in the MP and MP + GSPE groups decreased significantly by 24.14% and 13.52%, respectively (p < 0.05). Administration of MP resulted in significant increases in serum concentrations of blood urea nitrogen (p < 0.001), glucose (p < 0.01), alkaline phosphatase, and adrenocorticotropic hormone (p < 0.05). High-dose MP administration significantly reduced catalase (p < 0.001) and glutathione peroxidase (p < 0.05) concentrations in the liver and kidney tissues of rats, while glutathione concentrations were only reduced in liver tissue (p < 0.05). The expression levels of Bcl-2 and TNF-α in liver, kidney, and testicular tissue were significantly increased, while the expression levels of caspase-3 were reduced (p < 0.001). Furthermore, sperm concentration was significantly affected by GSPE in rats induced by high-dose MP, and sperm loss was significantly reduced in MP + GSPE (p < 0.05). These findings suggest that GSPE could be useful as a supplement to alleviate MP-induced toxicity in rats.

Circulatory cytokines during the piroplasm stage of natural Theileria annulata infection in cattle.

The objective of the present study was to investigate the inflammatory and anti-inflammatory cytokine response from a broad perspective in cattle with natural Theileria annulata infection. Ten cattle naturally infected with T. annulata and eight healthy cattle were included in this study. A total of 11 cytokines, including tumour necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin (IL)-1ß, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, and IL-17 were evaluated in serum samples using commercially available enzyme-linked immunosorbent assay (ELISA) kits. There was no statistical significance for serum TNF-α, IL-1ß, IL-5, IL-6, and IL-17 levels between the T. annulata infected and healthy cattle. In contrast, the median serum levels of IFN-γ (p = .023), IL-2 (p = .066), IL4 (p = .0016), IL-10 (p = .00087), IL-12 (p = .00018), and IL-13 (p = .023) were significantly higher in T. annulata-infected cattle than in healthy cattle. The results of the present study revealed that in the intraerythrocytic stage of tropical theileriosis, a very pronounced anti-inflammatory response occurs as well as an ongoing inflammatory process.

Effect of hydrated C60 fullerene on lipid, vitamin and amino acid composition in frozen-thawed ram semen.

This study was conducted to investigate the effect of different doses of hydrated C60 fullerene (C60HyFn) on freeze-thawing process-induced changes in lipid, vitamin and amino acid composition and also in motility, kinematic, sperm quality and oxidative stress parameters in ram semen. Semen was collected from seven rams twice a week for 3 weeks, so six repetitions were performed. The semen collected in each repetition was pooled. Each pooled sample was diluted with tris + egg yolk extender with (200 nM, 400 nM, 800 nM, 1 µM and 5 µM) and without (control) C60HyFn and they were frozen in mini straws. The doses of 800 nM, 1 µM and 5 µM had higher total, progressive motility, sperm membrane functionality rates, glutathione-peroxidase and catalase activities. All doses of C60HyFn significantly reduced dead and total abnormal sperm rates and malondialdehyde levels. Significant increases in vitamin A (400 and 800 nM doses), vitamin K1 (400 nM, 800 nM and 1 µM doses), total amino acid (all doses) levels, but significant decreases in vitamin D2 (800 nM, 1 and 5 µM doses), vitamin D3 (1 and 5 µM doses) and vitamin E (200 nM, 1 and 5 µM) levels were observed compared to control. In conclusion, the addition of C60HyFn to ram semen at 200 nM - 5 µM range, especially at a dose of 800 nM, provides a positive contribution to the protection of motility, vitamins A, K and total amino acid levels, and oxidant/antioxidant balance after freeze-thawing.

Effect of Hydrated Carbon 60 Fullerene on Frozen Ram Semen Quality.

The aim of this study was to evaluate the effect of hydrated carbon 60 fullerene (C60HyFn) on ram semen quality during cryopreservation. Three ejaculates from each of seven Akkaraman rams were collected using an artificial vagina during the nonbreeding season and pooled. Pooled semen samples were divided into 10 equal parts and diluted with tris + egg yolk extender not containing (control) and containing 100, 200, 400, and 800 nM and 1, 5, 10, 20, and 40 µM C60HyFn at 37°C. After addition of 5% glycerol and an equilibration process for 3 hours, the samples were frozen in 0.25-mL straws in an automatic freezing device at -140°C and stored in a liquid nitrogen container. Straws were thawed 24 hours after freezing and analyzed immediately with no incubation period. Motility, kinematic parameters, abnormality, vitality, hypo-osmotic swelling test (HOST), and oxidative stress levels were analyzed in thawed semen. Compared with the control, 200, 400, and 800 nM and 1 and 5 µM C60HyFn doses increased motility and HOST values and decreased the dead sperm rate. When compared with the control, addition of C60HyFn significantly decreased malondialdehyde levels (between 200 nM and 40 µM doses) and significantly increased glutathione peroxidase (between 800 nM and 40 µM doses) and catalase (between 1 and 40 µM doses) activities. In conclusion, results of this study show that the C60HyFn nanoparticles are nontoxic to ram semen and their supplementation in the extender is beneficial to sperm motility and membrane integrity after freeze-thawing.

Effect of freeze-thawing process on lipid peroxidation, miRNAs, ion channels, apoptosis and global DNA methylation in ram spermatozoa.

This study was carried out to investigate the effect of the semen freeze-thawing process on the functionality and molecular structure of ram spermatozoa. The temperature of pooled and diluted semen at 38°C (group 1, control) was lowered to 5°C (group 2), and it was subjected to glycerolisation-equilibration (group 3), frozen and thawed (group 4). Compared to the control, deterioration in spermatological parameters and significant increases in lipid peroxidation and global DNA methylation levels were observed in groups 3 and 4. When compared with the control, significant downregulation in the levels of miR-485 of group 2, miR-29a of group 3 and let-7a, miR-485 and miR-29a of group 4, and significant upregulation in the levels of miR-107 of group 3 and miR-127 of groups 3 and 4 were detected. In comparison to the control, significant upregulation in the levels of CatSper1, CatSper2, CatSper3, CatSper4, ANO1 and TRPM3 of group 2, CatSper4, ANO1 and TRPM3 of group 3 and KCNJ11 of group 4, and significant downregulation in the CatSper 3 level of group 4 were determined. As a result, the semen freeze-thawing process causes motility and morphological disorders in rams. This may be due to molecular changes associated with lipid peroxidation in spermatozoa.

The effect of vitamin E treatment during preovulatory period on reproductive performance of goats following estrous synchronization using intravaginal sponges.

The objectives of this study were to investigate whether the use of intravaginal sponge for estrous synchronization of goats causes oxidative stress, and to examine the effect of administering vitamin E during preovulatory period on reproductive performance of estrous synchronized goats. Estrus was synchronized in 36 non-lactating adult does using intravaginal sponges containing 30 mg of fluorogestane acetate (FGA) for 14 days. All females received 500 IU of eCG at the sponge withdrawal. The goats were allocated at random to two groups balanced for breed, age and body weight. Treatment group (n=18) received 200mg of vitamin E i.m. at the time of sponge removal and again at the time of second artificial insemination. The other 18 goats (control) were administered 1 ml of physiological saline instead of vitamin E on each of these two occasions. All does in estrus was intracervically inseminated at 12 and 24h after the onset of estrus. Blood samples were collected every 72h during the experimental period for evaluation of malondialdehyde (MDA) and vitamin E concentrations. Serum MDA level increased and vitamin E concentration decreased during the period of vaginal sponge application. Following the sponge removal, MDA level declined rapidly to below basal level in the treatment group but remained high in the control group. Conversely, vitamin E concentration increased in the treatment group after the sponge withdrawal and remained at a low level in the control group. No statistically significant differences (P>0.05) were observed between groups in terms of estrous response, conception rate, gestation length or kidding rate. However, the number of multiple births (70.0% versus 50.0%) and prolificacy rate (2.40+/-0.37 versus 1.63+/-0.26 kids per kidding) were significantly higher (P<0.05) for the treatment group than those of the control group. The results indicate that the use of intravaginal sponges for estrous synchronization of goats causes an increase in level of oxidative stress. However, the vitamin E treatment during preovulatory period can prevent the overproduction of reactive oxygen species (ROS), and it may improve the multiple birth rates and the number of kids born in estrous synchronized goats.