Japanese encephalitis virus (JEV) envelope (E) protein holds great promise for use in the development of a recombinant vaccine. Purified recombinant E (rE) protein may be useful for numerous clinical applications; however, there are limitations in using the Escherichia coli expression system for producing high-quality rE protein. Therefore, in this study, the yeast expression system was used to generate the rE protein. For protein production using the yeast system, the full-length JEV E gene was cloned into Pichia pastoris. SDS-PAGE and immunoblotting analysis demonstrated that the rE protein had a molecular mass of 58 kDa and was glycosylated. The predicted size of the mature unmodified E protein is 53 kDa, suggesting that post-translational modifications resulted in the higher molecular mass. The rE protein was purified to greater than 95% purity using combined ammonium sulfate precipitation and a SP-Sepharose Fast Flow column. This purified rE protein was evaluated for immunogenicity and protective efficacy in mice. The survival rates of mice immunized with the rE protein were significantly increased over that of Hyphantria cunea nuclear polyhedrosis virus E protein (HcE). Our results indicate that the rE protein expressed in the P. pastoris expression system holds great promise for use in the development of a subunit vaccine against JEV.
Objective: This study was performed to assess bacterial distribution concerned in sanitation management of public lavatory in Seoul. Methods: In this research, bacteriological investigation on public lavatory was accomplished for bidet water, bidet nozzle, washbowl and lavatory stool in the 50 public facilities such as public institutions, subway stations, cinema, department stores, large-scale buildings and hospitals amount to 374 specimens. Results: The geometric mean of colony forming unit(CFU) in total aerobic colony count were analyzed as follows; 5.2 × 10 2 /100 cm 2 on lavatory stool, 7.2 × 10 3 /ea on bidet nozzle, 7.8 × 10 3 /ea on center ring of washbowl, 1.4 × 10/mL in bidet water (ml) and 7.0/ea on doorknob. Opportunistically pathogenic germs such as Staphylococcus aureus, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginosa were isolated in 3.7%, 5.9%, 3.2% and 1.9% of total specimens, respectively. Conclusion: The result of this study shows that there were some facilities where the pathogenic germs were detected to may cause urological infection. And the CFU of general bacteria as the representative indicator of disinfection and lavatory cleaning were high enough to imply the improvement of sanitation management of public lavatories should be contrived.
Purpose : We will develop the distribution method of wide oil extract technology to develop repellent using this technology and study mosquito repellent persistence. Research design, data and methodology : A positive control group containing purified water, ethanol and picaridin was prepared, and the experimental control group was prepared in the same proportion as the positive control group, and 0.6% of broad oil was added. The results were summarized using the calculation method according to the avoidance effect and statistically tested by t-test using the excel statistics program. Results : Experiments on skin surface area and voice control of participants showed that men had 8.9% wider skin surface area than women, and voice control tests showed that women were bitten by mosquitoes five times more than men. Both the positive and the experimental control groups had a valid duration of up to three hours, but from the time of five hours, the positive control group had 77% and the experimental control had 90%, indicating a difference of 14.4% over the positive control group. Conclusions : The mosquito repellents developed in this study on the basis of safety and continuity are cost-effective in terms of mosquito repellent, and in addition fragrance, odor removal, perfume, ink, skin care and massage effect. .
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