배양계배 근원세포의 분화에 미치는 Fe3$^+$의 영향

유병제,지승완 한국통합생물학회 1991 동물학회지 Vol.34 No.4

계배 근원세포의 분화 및 증식에 미치는 F3+의 영향을 조사하였다. 철이 없는 배양액은 근원 세포의 분화와 증식을 억제하는 것으로 나타났으며 , 따라서 근원세포의 분화에 철과 transferrin (Tf)은 필수적이다. 또한, 철 대신에 Co2+가 부착된 Tf가 첨가된 배양액에서도 근세포의 분화가 정상적으로 일어나는 것으로 나타났다. Lysosomotrophic amine(chloroquine, $\mu$M 수준;ammonium chloride, mM 수준)은 근세포의 분화와 증식을 억제시켰으며 , 근세포 분화의 철에 대한 의존성은 분화됨에 따라 둔화되었다. 세포내로의 T니 수송량은 MEM과 8102배양액에서 비슷하였고, 근세포가 분화됨에 따라 감소하였다. Lysosomotrophic azine은 최소한 3시간이 내에서는 세포내로 수송되는 Tf의 양에 영향을 미치지 못하였다.

筋細胞 分化에 미치는 Fe³+의 影響

유병제,지승완,강봉석,양재섭 大邱大學校附設 基礎科學硏究所 1991 基礎科學硏究 Vol.8 No.-

Effects of Fe^(3+) on the differentiation and the proliferation of skeletal muscle cells in vitro were studied. It seemed that iron should be essential for the proliferation and the differentiation of myoblasts. Myoblasts cultured in the medium containing Tf added Co^(2+) instead of Fe^(3+) were normally differentiated.Lysosomotrophic amines inhibited the differentiation and the proliferation of chicken embryonic myoblasts in culture. The concentration required for inhibition of fusion were approximately 10μM of chloroquine, and 10mM of ammonium chloride. As myoblasts were differentiated, essentiality of iron for the differentiation was decreased.

Sucrose density gradient에 의해 분리된 C형 간염 바이러스의 특성 분석(Ⅱ)

황태욱,유병제 大邱大學校附設 基礎科學硏究所 2000 基礎科學硏究 Vol.16 No.2

To study the buoyant density of HCV, the human patient sera was tested with both anti-HCV ELISA and RT-PCR for the detection of HCV. The patient sera, positive at two tests, were used for further experiments. After ultracentrifugation of 20-60% sucrose linear density gradient, the centrifugal medium containing HCV was fractionated by 280 ul from the bottom. The buoyant densities of all fractions were determined, and the HCV in all fractions were checked by the RT-PCR. There was the room to separate the fractions to two major parts containing HCV. One was the high density part (1.146 g/ml ∼ 1.207 g(ml), and the other was the low density part (1.089 glml ∼ 1.108 glml).

Sucrose density gradient에 의해 분리된 C형 간염 바이러스의 특성 분석(Ⅲ)

황태욱,유병제 大邱大學校附設 基礎科學硏究所 2000 基礎科學硏究 Vol.16 No.2

To study the buoyant density of HCV, the human patient sera was tested with both anti-HCV ELISA and RT-PCR for the detection of HCV. The patient sera, positive at two tests, were used for further experiments. After ultracentrifugation of 20-60% sucrose linear density gradient, the centrifugal medium containing HCV was fractionated by 280 ul from the bottom. The buoyant densities of all fractions were determined, and the HCV in all fractions were checked by the RT-PCR. There was the room to separate the fractions to two major parts containing HCV. One was the high density part (1.131 g/ml - 1.240 g/ml), and the other was the low density part (1.103 g/ml - 1.122 g/ml). There was not the high density part in the serum treated with PAS for removing of antibody bound HCV. Therefore, it was concluded that the HCV in high density portion were the antibody bound forms, and the HCV in the low density portion were the free forms.

Anti-sense RNA에 의한 C형 간염 바이러스의 번역의 억제

지승완,유병제 大邱大學校附設 基礎科學硏究所 1998 基礎科學硏究 Vol.14 No.2

The gene therpys have been investigated as the new therpy of the desease, the inhibition of the specific gene expressin by anti-sense RNA is an one method of the them. The inhibition of viral translation or viral replication by anti-sense RNA causes the viral proliferation. Since hepatitis C virus is RNA virus, HCV may be the target virus of anti-sense RNA as anti-viral reagent. The inhibition of the translation of the defective virus having CAT as the reporting gene by anti-sense RNA was performed. The inhibition of CAT translation did not occure in cells pretransfected with anti-sense RNA, the inhibition of CAT tranalation occured in cells co-transfected with anti-sense RNA. It seems that the difference between pretransfection and co-transfection was resulted from the cellular delivery system of anti-sense.

Sucrose density gradient에 의해 분리된 C형 간염 바이러스의 특징 분석(Ⅰ)

황태욱,유병제 대구대학교 기초과학연구소 2000 基礎科學硏究 Vol.16 No.2

To study the buoyant density of HCV, the human patient sera was tested with both anti-HCV ELISA and RT-PCR for the detection of HCV. The patient sera, positive at two tests, were used for further experiments. After ultracentrifugation of 20-60% sucrose linear density gradient, the centrifugal medium containing HCV was fractionated by 280 ul from the bottom. The buoyant densities of all fractions were determined, and the HCV in all fractions were checked by the RT-PCR. There was the room to separate the fractions to two major parts containing HCV. One was the high density part (1.161 g/ml - 1.192 g/ml), and the other was the low density part (1.085 g/ml - 1.117 g/ml).

C형 간염 바이러스의 세포배양(Ⅰ)

황태욱,유병제 大邱大學校附設 基礎科學硏究所 2000 基礎科學硏究 Vol.17 No.2

In this research, It was investigated to increase the infectious efficacy of HCV to culture cells, and the efficacy of HCV cell culture system. To increase the infectious efficacy to human cells, The human hepatoma cell lines (Huh_(7),) was used as the culture cells to test the HCV infection. The whole patient serum, the high density portion, and the low density portion were added in the culture media of human hepatoma (Huh_(7)) cells as the infectious sources. It was showed that HCV partcles of the high density portion were not infectious to Huh_(7) cells, but HCV particles of the low density portion were infectious to Huh_(7) cells. Therefore, it was concluded that the antibody bound HCV was not infectious to Huh_(7) cells, but the antibody free HCV was infectious to Huh_(7) cells.

C형 간염 바이러스의 세포배양(Ⅱ)

황태욱,유병제 대구대학교 기초과학연구소 2000 基礎科學硏究 Vol.17 No.2

In this research, It was investigated to increase the infectious efficacy of HCV to culture cells, and the efficacy of HCV cell culture system. To increase the infectious efficacy to human cells, The human hepatoma cell lines (Huh_(7)) was used as the culture cells to test the HCV infection. The whole patient serum, the high density portion, and the low density portion were added in the culture media of Huh_(7) cells as the infectious sources. In the high density portion, the HCV genomes were not detected in the media and cells of all days except in the 2 day culture medium, but in the low density portion, the HCV genomes were detected in the media and cells of all days. It was showed that HCV partcles of the high density portion were not infectious to Huh_(7) cells, but HCV particles of the low density portion were infectious to Huh_(7) cells. Therefore, it was concluded that the antibody bound HCV was not infectious to Huh_(7) cells, but the antibody free HCV was infectious to Huh_(7) cells.

C형 간염 바이러스의 복제에 관한 연구

지승완,유병제 大邱大學校附設 基礎科學硏究所 1998 基礎科學硏究 Vol.15 No.1

Hepatitis C virus (HCV) is the major causative agent of posttransfusion and sporadic non-A, non-B hepatitis. To time, many research for HCV have been performed, but the molecular mechanism of HCV replication is still not obvious. To define the essential region of HCV genome for replication and develop the RNA-dependent RNA polymerase(RdRp) of HCV assay system in vivo. I constructed defective HCV cDNA that contained 5'Untranslate reagion,(UTR), Core(C), Enveloper 1(4 amino adds), Nonstructure(NS)5b, and various 3'UTR.

긴 cDNA를 이용한 C형 간염 바이러스의 검색

지승완,유병제 大邱大學校附設 基礎科學硏究所 1998 基礎科學硏究 Vol.14 No.2

Generally, the detection of virus was performed at the level of viral proteins and viral genome. The detection of virus by the viral genome more directly identifies the virus and estimates the concentration of virus. Because the genome of hepatitis C virus is RNA, the detection of the HCV genome was done by RT-PCR. But, PCR has the problems in contamination and false positive result. To deduce the upper problems, we developed the RT-PCR of the two ends of the HCV genome by long range cDNA. It seems that this method deduce the problems of contamination and false positive result, identify the full length genome of HCV and estimate the accurate concentration of HCV, and the fidelity and sensitivity of this method are good.