[ effects of Selenium on changes in sperm antioxidant capacity in old and adult rats]

Selenium as an antioxidant is essential for normal function of testis and spermatogenesis. It can reduce formation of free oxygen radicals and as a result it is expected to improve male fertility. The aim of this study was to evaluate alterations in antioxidant capacity of old rats sperms after prescription of 0.2 mg/kg of Selenium. In this study, 15 old male rats of 10-12 months of age and 15 adult male rats of 2-3 months of age were randomly divided into three groups: control, sham and experimental groups. Control group did not receive any treatment; sham group rats received intra peritoneal injections of equal volume of Selenium solvents [normal saline] as Selenium in experimental group. Experimental groups of male rats received daily intraperitoneal injection of Selenium [0.2 mg/kg] for 5 weeks. After 42 days from initiation of injection, the rats were killed by cervical dislocation and after obtaining sperm, total antioxidant capacity of the sperms was measured by FRAP assay. The absorbance of TPTZ-fe[+2] was read at 593 nm by spectrophotometery. For the statistical analysis, SPSS software was used and data analysis was performed by means of Kruskal-Wallis and Mann-Withney U tests. P<0.05 was considered significant. Results of this study showed significant differences in mean values of total antioxidant capacity in both old and adult rats in experimental and control groups [P<0.05]. Also comparison of mean values of antioxidant capacity of sperm solution in adult and old control groups showed a significant difference [742.26 +/- 1.06 vs. 672.061 +/- 0.78 respectively] [P<0.05]. The results of this study showed that Selenium treatment in old rats [0.2 mg/kg after 35 days] can improve total antioxidant capacity of the sperms of old rats. Regarding low levels of antioxidants in old rats, it can be suggested oxidative stress can result in diminution of antioxidant levels. Therefore antioxidant therapy could be considered as a method for improvement of the quality of sperms of old men

Evaluation of the efficiency of pIRES2 EGFP and pcDNA3-hBDNF-v5 plasmids in transfection of CCE ES cells by the electroporation method

Unlimited self renewal and potential capacity of embryonic stem cells [ESCs] in differentiating into a wide variety of cell types has made the cells an attractive source of donor cells for developmental studies and cell therapy. The aim of the present study was the evaluation of the transfection efficiency of pIRES2-EGFP and pcDNA3-hBDNF-v5 plasmids in CCE ES cells by the electroporation method. The plasmids transformed into DH5

Sex determination of human fetus using nested-PCR

One of the goals of modern genetics is to develope safe prenatal diagnostic tests which do not constitute any risk to the fetus. One potentially non-invasive approach for doing so is to obtain fetal material from the maternal circulation. In this study we obtained maternal blood from the pregnant women and tried to determine sex of fetus by nested-PCR. To determine the sensitivity of the PCR methods, artificial samples were prepared first by mixing whole blood of adult males with that of the adult females. After performing DNA extraction, PCR was optimized on these samples. Whole blood samples were then obtained from 70 pregnant women in 8 to 12 weeks of gestation after considering the medical ethics. In this study we tried to extract the DNA of the white blood cells [WBC] as well as free DNA of serum and plasma of the maternal bloods and performed Nested-PCR using primers flanking the specific-sequence of Y-chromosome or Sex determining region on Y [SRY]. If the fetus was male, the size of amplified fragment of the first round product was about 470 base pairs and second round was 254 base pairs. After delivery, Only 32 out of 70 of the samples could be followed and results showed that 18 were male. However, the results of the nested-PCR examination of the serums indicated that 21 of the fetas were male and examination of the WBCs showed that 22 were male, in each group 3 and 4 cases were misinterpreted respectively. So the accuracy of sex determination of the samples in serums and WBCs were 90.62% and 81.25% respectively which is almost similar to the recent reports [91.5%]. We could not obtain good results from the free DNA of the plasma. We have obtained relatively positive results from the analysis of maternal blood and if we could follow all cases to see the results of all deliveries, we could then reconfirm the results of our research. Anyway we hope this project could be able to introduce this new approach as means of noninvasive prenatal diagnosis for detection of common inherited diseases in Iran

Evaluation of Catsper gene expression in mouse testis at different ages

Recently, a novel gene was cloned and characterized namedCatSper, which codes for a unique Ca2+ channel that is expressedexcludively in the testis. It has crucial role in sperm motility, spermpenetration into the ovum and ultimately male fertility. In the present study we have evaluated the expression of this gene at different ages of mouseby Semi-quantitative RT-PCR. Material and Total RNA was extracted from testis of mouse usingRNX plus solution. Reverse Transcription was done by oligo dT and MMLVenzyme. cDNAs were amplified in PCR reaction for both CatSper and b2m[as an internal control] genes. After gel electrophoresis, intensity of signalfor each band was quantified by Labimage software. 1] There was no d expression of CatSper gene in mouse testis of1 day, 1 week and 2 weeks [pre-mature group] of age;2] Expression ofCatSper began with the age of three weeks with a gradual increase till twomonths of age and a gradual slight decrease in older age groups;however, the observed pattern of expression appeared not to bestatistically significant [p=0.592]. These results confirm previous data; that is there is a linkbetween expression of CatSper and fertilization in mouse. The resultsfurther reveal a link between CatSper expression profile and the age ofmouse in terms of sexual maturity