Slide Session : OS-IFD-07 ; Infectious Disease : In Vitro Antiviral Activity of Ribavirin Against Severe Fever with Thrombocytopenia Syndrome Virus

( Myung Jin Lee ),( Kye Hyung Kim ),( Jong Youn Yi ),( Su Jin Choi ),( Chung Jong Kim ),( Nak Hyun Kim ),( Kyoung Ho Song ),( Pyoeng Gyun Choi ),( Ji Hwan Bang ),( Wan Beom Park ),( Eu Suk Kim ),( San 대한내과학회 2014 대한내과학회 추계학술대회 Vol.2014 No.1

In Vitro Antiviral Activity of Ribavirin Against Severe Fever with Thrombocytopenia Syndrome Virus Myung Jin LEE1, Kye-Hyung KIM1, Jongyoun YI2, SuJin CHOI1, Chung-Jong KIM1, Nak- Hyun KIM1, Kyoung-Ho SONG1, Pyoeng Gyun CHOI1, Ji-Hwan BANG1, Wan Beom PARK1, Eu Suk KIM1, Sang-Won PARK1, Hong Bin KIM1, Nam Joong KIM1, Myoung- Don OH1 Seoul National University College of Medicine, Korea1, Pusan National University School of Medicine, Korea2 Background: Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease caused by a novel Bunyavirus, severe fever with thrombocytopenia syndrome virus (SFTSV). No effective antiviral therapy is proven yet, but clinical use of ribavirin (RBV) has been tried. We investigated the antiviral effect of RBV against SFTSV in vitro. Methods: To test for cytotoxicity of RBV, Vero cells were treated with different concentrations of RBV (3.90 to 500 μg/mL, two-fold dilution) and analyzed by cell viability MTS assay 48h post-infection. To determine antiviral activity of RBV against SFTSV, Vero cells were infected with SFTSV strain Gangwon/Korea/2012 at 100 TCID50 (50% tissue culture infective dose) per well in a 96-well plate, and RBV was added at the concentrations showing no or minimal cytotoxicity. Viral RNAs were extracted from the culture supernatants and quantifi ed using one-step real-time reverse transcription- PCR to amplify the partial large segment of SFTSV. Statistical analysis was done by one-way ANOVA with Tukey`s post hoc test. Results: Cytotoxicity due to RBV was not observed at RBV concentration =31.3 μg/ mL. Viral RNAs at 24h post-RBV treatment were reduced with increasing RBV concentrations (1-32 μg/mL), compared with those of mock-treated cells (P <0.01, Figure). Half maximal inhibitory concentration (IC50) of RBV was 3.69 μg/mL at 24h post-RBV treatment. Conclusions: Our study shows that RBV has antiviral effect against SFTSV in a dose-dependent manner. Further studies are required to evaluate the effi cacy of RBV in SFTS.

Tunicamycin에 依한 枯草菌 細胞外 酵素分泌機構의 解析

宋邦鎬 경북대학교 과학교육연구소 1981 科學敎育硏究誌 Vol.5 No.-

고분자단백이 세포외로 분필되는 기구는 세포질막에 결합된 ribosome에서 합성되어 cotranslational transport 즉 합성과 동시에 분필된다고 믿어지고 있으며 이때 세모막을 통과하기 위한 기구로서 signal peptide 합성에 의한 signal hypothesis와 cytoplasm에서 합성된 단백의 3차구조 변형에 의한 membrane trigger hypothesis가 있다. 이와같은 단백의 분필기구를 해명하기 위해 G. Blobelletal은 일찍이 eukaryotic cell에서의 단백분필에 대한 model을 발표하였으며 관련분야 연구자들에 의하여 IgG ight chain, insulin, ovomucoid, lysozyme 등에서 signal peptide를 확인 하였다. 또 prokaryotic cell에서는 대장균의 lipoprotein, maltose binding protein, λ-receptor, alkaline phosphatase 등에서 가장 연구가 많이 수행되어왔다.

탄화수소 자화세균의 분리

宋邦鎬,河惠貞 慶北大學校 1989 論文集 Vol.47 No.-

Four strains of hydrocarbon utilizing bacteria from the sludges of petroleum factory were isolated by testing the growth abilities to utilize methanol, benzene, paraffin and petroleum. Each strain was designated as KM5001 ,5002 ,5003 and 5004, and the putative genus was identified as Neisseria, Flavobacterium, Streptococcus, and Pseuomonas, respectively. These strains were grown well in the CⅢ minimal medium supplemented with above hydrocarbon sources. The strains KM-5001 and 5002 were grown better than the others by utilizing various hydrocarbons and methanol. However, considering its doubling time and specific growth rates, all strains could be applicable for the industrial use for the purpose of removing the petroleum contamination from the sea and river. The strains were grown well from pH 6.0 to 7.0, while the growth rate was decreased at pH 9.0. The optimal growth temperature was 40C. Above40C, the growth rate of cells reduced extremely. All strains carried plasmids in which the genes for utilizing hydrocarbons may be inherent.

Salmonella typhimurium cad 오페론의 산성유도에 관여하는 CadR과 CadC의 역할

방성호 한서대학교 산학협력연구원 부설 환경연구소 1999 환경연구소 논문집 Vol.2 No.-

S. typhimurium cadBA operon의 발현에는 산선 pH와 고농도의 라이신등 적어도 두가지의 세포외부 신호가 요구된다. pH, 라이신 신호에 따른 cadBAqkfgus 조절기작은 이해하기 위하여 Tn10삽입, 자발적 돌연변이 EMS 돌연변이 등을 수행하여 JF2238과 cadA-lacZ의 발현특성에 차이를 보이는 돌연변이체를 선별하였다. cadR::Tn10돌연변이와 cadR3, cadR4 돌연변이는 산성 pH, 라이신이 없는 조건에서 cadA-lacZ 발현을 유도하였고, 라이신 독성 유사체인 thiosine에 대해 내성(Ts')을 나타냈으며, E. coli의 lysP 클론을 갖고 있는 pLYSP에 의해 돌연변이 형질이 보상되었다. 따라서, CadR은 라이신이 없는 조건에서 cadBA 발현에 음성적 조절자로서 작용함을 알 수 있었다. cadC6 돌연변이는 pH-비의존성, 라이신-비의존성 cadA-lacZ 발현을 보였고, cadC4, cadC5 돌연변이는 pH-비의존성, 라이신-의존성 발현을 나타냈고, 모두가 cadA와 연관되어 존재하였기 때문에 pH와 라이신 신호에 대해 반응하는 부위가 cadC 내에 있을 것이라 추정하고 있다. 또한 cadR3, cadR4는 cadR::Tn10과 마찬가지로 pH-의존성, 라이신-비의존성 cadA-lacZ 발현을 보여, cadR은 라이신 신호에 대해 작용하고 있었다. 그러나 cadR3가 cadC+ 상황에서만 cadA-lacZ 발현을 나타내기 때문에 pH 신호와 라이신 신호 모두가 CadC에 의해 매개되고 있을 것이라 예상하고 있다. 또한 LDC의 반응산물인 카데브린(cadeverine)은 cadC+ 균주에서 cadA-lacZ 바련에 억제효과를 나타냈으나, cadC 돌연변이체는 억제효과가 없었기 때문에 카데브린 신호도 CadC를 통해 전달되고 있었다. The expression of S. typhimurium cadBA operon requires at least two extracellular signals : low pH and high concentration of lysine. To further characterize nature of pH-dependent and lysine-dependent signal transduction, the mutants which express a cadA-lacZ operon fusion in various growth conditions by Tn10 insertion, spontaneous mutagenesis, and EMS treatment were isolated. Two cadR::Tn10, cadR3 and cadR4 mutants were isolated from which cadA-lacZ was expressed in the absence of exogenously added lysine. They were also resistant to thiosine and complemented by lysP clone from E. coli. Thus, in the absence of exogenous lysine, cadR is negative regulator of cadBA expression. The other mutation(cadC3 and cadC4) in cadC conferred pH-independent and lysine-dependent cadBA expression. Cadaverine, the product of lysine decarboxylation, was shown to inhibit expression of cadA-lacZ fusion in cadC+ cell.

부산광역시 하수관거의 효율적 확충방법에 관한 연구 : 남부하수처리구역을 중심으로 A Focus of Nambu Sewage Disposal Area

송방환,이성호 釜山大學校 都市問題硏究所 2003 都市硏究報 Vol.14 No.-

Sewer equipment is life equipment that take charge basic role for inundation protection and life environment improvement. this should be enough exhibition of the function to public water area's quality of water preservation and city prevention of disasters equipment. In such background, this research grasps problem of existing sewer equipment and establish improvement plan. And this research can do efficiently existent sewer institution to become expansion and maintenance. It is purpose of study.

腹水癌細胞의 凝集物質

宋邦鎬 慶北大學校 1979 論文集 Vol.27 No.-

Agglutination factor of carcinoma cells(Ehrlich carcinoma ascites) in mouse was detecrted from the cultured broth of actinomycete. About 120 strains of actinomycete were isolated from soil sample, and determined the producing ability of agglutinating factor. The agglutination activity of carcinoma cells was found to be directly proportional to it's proteolytic activity. Moreover, the carcinoma cells were agglutinated by the proteolytic enzyme itself; pepsin, trypsin and papain, whereas it was not coagulated by amylase. So, in the cultured liquid, the proteolytic enzyme may be responsible for the agglutination in the culture. Two species of Streptomyces were isolated as the most favorable strains for secreting the agglutinating component, Out of these, the strain with higher agglutinating activity was found to be stable in wide ranges of the pH and temperature. And it was recovered from the non-dialyzable fraction as a high molecular proteolytic substance.

信號假設 : 蛋白質의 生體膜 通過機構 The Mechanism of Protein Translocation through Biological Membranes

宋邦鎬 慶北大學校 1985 論文集 Vol.39 No.-

The mechanism of protein translocation by signal hypothesis, especially, the function of signal sequence and the genetic approach by mutation on the sequences, was studied for elucidating the signal role of the sequences on the initiation step of protein translocation. The signal peptidase, internal signal sequence, stop transfer sequence and sorting sequence were also described as regulatory segments for secretory proteins or for membrane-bound ones in relation to the topology in the cells. The cotranslational and posttranslational translocation were suggested in the protein translocation of eukaryotic and prokaryotic cells. The signal recognition particle and docking protein were isolated and characterized in the secretion system of eukaryotic cells. However, those were not recognized in prokaryotic cells. The hydrophobic segments of protruded nascent polypeptides in N-terminal of secretory proteins may have a role of easy attachment with an affinity to the hydrophobic membranes for forming a channel of transporting the extended polypeptides. Finally, the membrane triggers hypothesis as a post-translational transport system was reviewed. The mixing of the above two theories has created a possible new theory which has only been studied briefly.

Caprolactam 資化細菌의 分離 및 藥劑耐性

宋邦鎬 慶北大學校 師範大學 1983 敎育硏究誌 Vol.25 No.-

Seven kinds of ε-caprolactam utilizing bacteria were isolated from the cooling water of the nylon fibre in nylon manufacturing factory. One of them, designated as a strain MK-7 in our collections, was the most powerful strain for utilizing ε-caprolactam. Most of these bacteria were coccal or bacilli types and had flagella for motility. The mode of cell division was simple. These cells were colored albino or ivory white in agar plate and soluble pigments were not produced. Luphobos MK-11 and Spinnig A as a sterilizing agents were very effective to these bacteria. Under the concentrations of about 50 to 100 ppm, most of these strains were inhibited in the growth perfectly. And these bacteria were sterilized by UV for 15 seconds of irradiation.

Bacillus subtilis의 cdd유전자의 Escherichia-Saccharomyces Shuttle Vector YRp7에의 클로닝

金榮基,宋邦鎬 경북대학교 과학교육연구소 1988 科學敎育硏究誌 Vol.12 No.-

The cdd gene enconding cytidine deaminase from Bacillus subtilis was cloned and expressed in the Saccharomyces cerevisiae D13-1A. By insertion of the 1.45 Kb of EcoRl/EcoRl fragment which containing genes fo for replication origin and trpl selective marker from Escherichia-Sac-charomyces shuttle vector YR_p7 into the EcoRl site of pS062, the Bacillus cdd gene was expressed in the yeast cells. By selection of the trp+ colonies in the SD medium. One of the transformed Saccharomyces cells were isolated as Saccharomyces cerevisiae YKl. The transformed yeast cells which carring chimeric plasmid, pYKl, was expressed the cdd activity with two folds elevated levels. For further confirming the pYKl expression in yeast cell, the pYKl which isolated from the transformed cells was recloned into the Escherichia coli KM268. The plasmid isolated from strain KM268 was identified as the same one with the original chimeric plasmid, pYKI.

정하중 및 동하중을 받는 강구조물의 최적 설계에 관한 연구

노승훈,방중철,송호중 金烏工科大學校 1994 論文集 Vol.15 No.-

Nowadays, the parking is a very serious problem almost everyhere. It is even more serious in the parking lot of the high storied apartment. Many parking devices have been developed to solve this problem by increasing the parking capacity within the limited areas. In this study parking devices are considered to obtain the optimum for the capability of carrying the required load and for the minimization of the material usage for the structure. The design of the parking structure studied in this work can readily be applied to many different parking devices.