Effect of Mouse Ovarian Vitrification on Promoter Methylation of Inhba and Inhbb in Granulosa Cells of Follicles.

BACKGROUND: Cryopreservation can induce cellular, genomic, and epigenetic abnormalities. OBJECTIVE: To analyse the impact of ovarian vitrification on follicular development and its epigenetic effect on promoter methylation of Inhba and Inhbb in granulosa cells. MATERIALS AND METHODS: Mouse ovaries were divided into control, toxicity, and vitrified groups. The growth and development of follicles were examined. After in vitro culture of follicles, DNA was extracted from isolated granulosa cells and treated with sodium bisulfite. The promoter methylation of Inhba and Inhbb was analyzed by direct PCR sequencing. RESULTS: Vitrification reduced the growth of follicles; however, antral cavity formation was not influenced negatively. Vitrification reduced the percentage of 5-methylcytosine in the Inhba promoter, while CpG sites in the promoter of Inhbb remained unmethylated. CONCLUSION: Vitrification had adverse effects on follicle growth and the epigenetics of granulosa cells. The results of the current study show that vitrification methods of ovary need more improvement.

The effects of Cannabis sativa L. seed (hempseed) in the ovariectomized rat model of menopause.

Cannabis sativa L. has been used for the treatment of various gynecological diseases in traditional medicine. The potential of this plant to protect against complications of menopause has been raised but rarely studied. Twenty female rats were divided into five groups: sham-operated (sham), ovariectomized (OVX) and three other ovariectomized groups: HST1%, HST2% and HST10% which received 1%, 2% and 10% hempseed, respectively, in their diet for 3 weeks. The effects of hempseed on plasma lipid and lipoprotein profiles, estradiol and calcium levels were evaluated. Rats were tested for behavioral changes using the forced swimming test. The results showed that ovariectomy, independent of the type of diet, caused elevation of plasma calcium, total cholesterol and HDL-cholesterol levels, while hempseed modified this effect. Plasma estradiol levels were significantly lower in the OVX group compared to other groups. The swimming times for the OVX and sham groups were significantly shorter than that of the HSD10% group. All hempseed-treated groups were less anxious and showed significant declines in fecal boli compared to the sham group. The exploratory diving percent decreased in the HST10% group compared with other groups. These results suggest that hempseed may improve post-ovariectomy complications in rats.

Selective determination of ethyl acetate, acetone, ethanol, and methyl ethyl ketone using quartz crystal nanobalance combined with principle component analysis.

Quartz crystal nanobalance (QCN) sensors are considered as powerful mass sensitive sensors to determine materials in the subnanogram level. In the current study a method based on QCN modified with polyethylene glycol (PEG) has been developed to determine organic vapors (ethyl acetate, acetone, ethanol and methyl ethyl ketone). The frequency shift of QCN was found to be linear against analytes concentrations in the range between 4 to 35 mg/L for acetone vapor and 4-70 mg/L for 3 other vapors. The correlation coefficients for ethyl acetate, acetone, ethanol, and methyl ethyl ketone were 0.9971, 0.9976, 0.9984 and 0.9927, respectively. The principal component analysis was also utilized to process the frequency response data of the organic vapors. Using principal component analysis, it was found that over 95% of the data variance could still be explained by use of two principal components (PC1 and PC2). Subsequently, the successful discrimination of ethyl acetate and other compounds was possible through the principal component analysis of the transient responses of the PEG-modified QCN sensor.