Evaluating And Testing Of A Potential Dna Vaccine Against Vibrio Cholerae

Although it has been more than 100 years since the first attempt to produce a cholera vaccine was made, an effective cholera vaccine has yet to be developed. In this study, the level of protection produced by a potential DNA vaccine (PVax/ctxB) was tested against the ctxB toxin of Vibrio cholerae on Balb/c mice. First, the intramuscular vaccination method was validated using pCMV plasmid that encodes HbsAg, which was detected 5 days after the injection into the tibial muscle. Next, 4 groups of mice were intramuscularly injected with either the pVax/ctxB vaccine construct or pVaxl as the negative control. The first and second groups received 2 injections spaced 3 weeks apart, while the other two groups were given 3 injections spaced 3 weeks apart. This was then followed by challenging the mice with 105 or 107 cfu/ml/mouse from clinical isolates of V. cholerae after 3 weeks of the last injection. Antibody levels for both IgG and serum IgA were monitored using ELISA, and showed high production of IgG after the first booster injection with no significant change of IgA levels

Changes in Lactate Production, Lactate Dehydrogenase Genes Expression and DNA Methylation in Response to Tamoxifen Resistance Development in MCF-7 Cell Line

Lactate dehydrogenase (LDH) is a key enzyme in the last step of glycolysis, playing a role in the pyruvate-to-lactate reaction. It is associated with the prognosis and metastasis of many cancers, including breast cancer. In this study, we investigated the changes in LDH gene expression and lactate concentrations in the culture media during tamoxifen resistance development in the MCF-7 cell line, and examined LDHB promoter methylation levels. An upregulation of 2.9 times of LDHB gene expression was observed around the IC50 concentration of tamoxifen in treated cells, while fluctuation in LDHA gene expression levels was found. Furthermore, morphological changes in the cell shape accompanied the changes in gene expression. Bisulfate treatment followed by sequencing of the LDHB promoter was performed to track any change in methylation levels; hypomethylation of CpG areas was found, suggesting that gene expression upregulation could be due to methylation level changes. Changes in LDHA and LDHB gene expression were correlated with the increase in lactate concentration in the culture media of treated MCF-7 cells