Evaluation of the number of CD4[+] CD25[+] FoxP3[+] Treg cells in normal mice exposed to AFB1 and treated with aged garlic extract

Aflatoxin B[1] [AFB[1]] suppresses the immune system. To decrease such sup-pressive effects on the immune system, a wide range of herbal medicines like garlic are utilized. Biological activities of garlic in vitro and in vivo have also been verified. Our previous studies demonstrated that aged garlic [dry garlic bulbs preserved in the freezer for six months at -20°C] have increased immunostimulator fractions and reduced immunosuppressor fractions. This study focuses on the immunosuppressor activity of AFB[1] and immunostimulator activity of aged garlic extract [AGE] through the evaluation of CD4[+] CD25[+] FoxP[+] regulator cell [Treg] counts and the pattern of cytokine production in Balb/c normal mice. In this experimental research, AFB[1] was separated from Aspergillus flavus [PTCC 5004] by HPLC and AGE prepared using the Mantis method. The Delayed-Type Hypersensitivity [DTH] test was carried out to determinate the effectiveness of different doses of AGE and AFB[1], which can both have an effect on the immune system. Subsequent experiments were carried out on 20 Balb/c mice to estimate the effects of AGE and AFB.[1] on the number of Treg cell in 4 groups: 10 microl/kg/day of AFB [1], and AGE diluents were administered for 4 consecutive days to group 1. AFB[1], 2. control, 3. AGE + AFB[1] and 4. AGE via intraperitoneal [IP] route, respectively. Mice were sacrificed and splenocytes harvested and the percentage of splenic Treg cells was measured by flow cytometry analysis. The ELISA method was utilized to measure Cytokine production. The findings reveal that AGE increased the level of IFN-gamma and IL-4 cytokines produced by splenocytes stimulated by specific tumor antigen and decreased the number of Treg cells in the spleen [p<0.05]. AFB[1] increased the number Treg cells in the spleen and decreased cytokine production [p<0.05]. In groups 2 [control] and 4 [AGE] the number of Treg cells decreased [p value<0.05] whereas in groups 1 and 3 the number of Treg cells increased [p<0.05]. This study indicated that AGE is able to alter the cytokine production in normal mice into a Th[1] protective pattern which is beneficial to the immune system in general and anti-tumor immunity in particular. AFB.[1] is able to alter the cytokine production into a Th[2] protective pattern. Therefore, AGE might be used as herbal medicine with few side effects as compared to chemotherapy in treating cancers caused by substances like AFB[1]

[ effects of Taurine antioxidant on standard parameters, DNA fragmentation, Protamine deficiency and oxidative stress on freezing human semen]

Evaluation of the effects of different doses of antioxidant Taurine on oxidative stress and human sperm parameters following cryopreservation. The semen of 20 fertile men were divided to 5 aliquot, one part considered as a fresh after analysis of standard semen parameters [Motility, Abnormal Morphology, Viability] and Protamine deficiency, DNA damage and measurement of ROS and RNS. The other part was loaded on to a 80% and 40% Allgrad gradient and centrifuged. The pellet was washed and divided into 4 separate fractions for control [non-frozen] and cryopreservation groups in absence or presence of 0.25 and 50 mM Taurine. The frozen specimen were thawed and then examined. Sperm Motility evaluated using a CASA software. The Viability of spermatozoa was assessed by the Trypan-Blue stain method. Levels of ROS determined by spectroflorometry assay using DCFH-DA. DNA fragmentation examined by SCD test and Protamine deficiency examined by CMA3+ staining. At the end results were analyzed using ANOVA test. Cryopreservation procedure increased the amount of ROS and adding 25 mM of Taurine improved post-thaw motility, progressive motility and sperm protamine deficiency. However, different doses of Taurine [25 and 50 mM] had no significant effects in total abnormalities, viability, DNA fragmentation and ROS reduction. Antioxidant Taurine has no significant effects on ROS production following human sperm cryopreservation. But with dose of 25mM could improve the quality of spermatozoa due to the assessment of motility and protamine deficiency