소아의 연령별 Propionibacterium acnes에 대한 균체응집항체 및 동종혈구응집항체간의 비교연구

정화영,정상인,최철순,양용태 중앙대학교 의과대학 의과학연구소 1984 中央醫大誌 Vol.9 No.3

In general, the concentration of maternal IgG globulin in serum of the neonates falls rapidly within the first few months after birth and the production of IgM globulin develops in the maturing infants during the course of exposure to various antigens in the environment. The development of natural antibodies, ie., isohemagglutinins, IgM class, to ABO blood group substances and agglutinins, Ig class, to normal flora, in the early stage of life is important since not only they may act as bactericidal substances in nonspecific manners, but also could be immunological barometers on the normal function of humoral immune system. The high concentration of isohemagglutinins to human A or B blood group antigens and agglutinating antibodies to some of normal flora such as Propionibacterium acnes or Staphylococcus aureus were observed in normal human sera. It has been known that the serum concentration of IgM globulin usually reached adult levels by one year of age, while that of IgG globulins by five to six years of age. However, the levels of isohemagglutinins to A and B group substances and agglutinating to Propionibacterium acnes in children's sera and the ages in which the concentrations of their antibodies reached to abult levels are not clarified. In this study, the concentrations of isohemagglutinins to A and B blood group antigens and agglutinating antibodies to P.acnes serotype Ⅰ and serotype Ⅱ in the sera of 163 normal children, ranged from 0 day to 15 years of age, were measured by means of microtitration technique. The results obtained are as follows: 1. In the sera of 163 children under 15 years of age, there observed no significant difference in the titers of agglutinating antibodies to P. acnes serotype Ⅰ andⅡ. 2. Of 75 sera of children under one year of age, the numbers of sera in which agglutinating antibodies were not detectable or less than 1:4 to P. acnes serotype Ⅰ were 67(89.6%) and to serotype Ⅱ 53(70.7%), respectively. 3. Agglutinating antibody to P. acnes serotype Ⅰ in the children's sera reached adult levels by 7 years of age, but 100 percentages of antibody detection was observed only in the age group of 15 years old, whereas the adult levels of isohemagglutinins to A and B blood group antigens were observed in the age group of 6 months old. 4. No correlation were observed in normal children's sera between agglutinating antibody titre to P. acnes serotype Ⅰ and isohemagglutinin titre. These results indicated that the isohemagglutinins to ABO blood substances appeared in the earlier stage of life than did agglutinating antibodies to P. acnes.

대장균군 검사용 간이 시험지 개발

이인애,김재화,이희구,성창근,최인성,정태화 충남대학교 생물공학연구소 1998 생물공학연구지 Vol.6 No.-

대장균군 검사용 간이 시험지는 본 실험실에서 국내 최초로 고안, 개발하였으며 이 간이 시험지법은 현장 검사법의 하나로 대장균군이 내는 succinic acid dehydrogenase 때문에 tetrazolium salt 가 환원되어 적색 반점을 형성하는것을 이용한 방법으로서 이 간이 시험지의 제조는 대체로 종래의 표준 평판법과 거의 동일한 조성의 배지와 시약을 사용하여 여지에 흡착시킨 후, 건조시켜 (60℃) 멸균한 것으로 표준 평판법과 어떤 상관관계가 있는가를 검토하였다. 이 간이 시험지의 제조에서는 bile salt No. 3를 deoxycholate로 대체하여 제조 원가를 절감하였고, 또한 일본에서 현재 시판되고 있는 제품과 품질 비교시험을 하여 더 좋은 결과를 얻었으며 종래의 표준 평판법과 비교하였을 때도 오히려 표준 평판법(24-48시간 배양)보다 빠른 시간(16-20시간 배양)내에 판정할 수 있는 이점이 있으며, 표준 평판법에서는 없어서는 안될 배지나 배양 접시, pipette등의 자료및 기구가 일체 필요없고 언제 어디서나 현장에서 직접 시험할 수 있어 매우 간편하며 또한 저렴한 가격으로 제조할 수 있는 경제성이 높은 이점을 갖고 있다. The objective of this study was to develop a paper strip which could determine E. coli qualitatively and quantitatively in water, wastewater, drinks, or food. This paper strip method was a simple and rapid test method that determine E. coli by visual identification. In this study, nutrient culture media were formulated and characterized for optimum conditions. Paper strips were then prepared by impregnating into the media and dried at 60℃. The test procedure is quite simple to use. The paper strip was dipped into a sample, and excess sample was removed. The strip was then incubated at 37℃ for 16 to 20 hours and the number of colonies on the strip was counted. The color of the colony spots produced by microorganisms varied depending on the media formulation. Violet-red spots were produced by E. coli. The test method was simple, rapid and no special laboratory equipment was necessary for visual identification. Therefore, this test method is applicable to on-site tests such as field tests or home tests. The paper strip method was compared with the standard agar plate method and Japanese commercial product. The method of the economical preparation of test strips was studied for production on industrial scale.

대장균군 검사용 간이 시험지 개발

이인애,정태화,김재화,성찬근,이희구,최인성 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1996 Journal of biomedical laboratory sciences Vol.2 No.1

대장균군 검사용 간이 시험지는 본 실험실에서 국내 최초로 고안, 개발하였으며 이 간이 시험지법은 현장 검사법의 하나로 대장균군이 내는 succinic acid dehydrogenase 때문에 tetrazolium salt 가 환원되어 적색 반점을 형성하는것을 이용한 방법으로서 이 간이 시험지의 제조는 대체로 종래의 표준 평판법과 거의 동일한 조성의 배지와 시약을 사용하여 여지에 흡착시킨 후, 건조시켜 (60℃) 멸균한 것으로 표준 평판법과 어떤 상관관계가 있는가를 검토하였다. 이 간이 시험지의 제조에서는 bile salt No.3를 deoxycholate로 대체하여 제조 원가를 절감하였고, 또한 일본에서 현재 시판되고 있는 제품과 품질 비교시험을 하여 더 좋은 결과를 얻었으며 종래의 표준 평판법과 비교하였을 때도 오히려 표준 평판법(24-48시간 배양)보다 빠른 시간(16-20시간 배양)내에 판정할 수 있는 이점이 있으며, 표준 평판법에서는 없어서는 안될 배지나 배양 접시, pipette등의 자료및 기구가 일체 필요없고 언제 어디서나 현장에서 직접 시험할 수 있어 매우 간편하며 또한 저렴한 가격으로 제조 할 수 있는 경제성이 높은 이점을 갖고 있다. The objective of this study was to develop a paper strip which could determine E. coli qualitatively and quantitatively in water, wastewater, drinks, or food. This paper strip method was a simple and rapid test method that determine E. coli by visual identification. In this study, nutrient culture media were formulated and characterized for optimum conditions. Paper strips were then prepared by impregnating into the media and dried at 60℃. The test procedure is quite simple to use. The paper strip was dipped into a sample, and excess sample was removed. The strip was then incubated at 37℃ for 16 to 20 hours and the number of colonies on the strip was counted. The color of the colony spots produced by microorganisms varied depending on the media formulation. Violet-red spots were produced by E. coli. The test method was simple, rapid and no special laboratory equipment was necessary for visual identification. Therefore, this test method is applicable to on-site tests such as field tests or home tests. The paper strip method was compared with the standard agar plate method and Japanese commercial product. The method of the economical preparation of test strips was studied for production on industrial scale.

미립자 응집반응을 이용한 C-reactive Protein의 면역측정법에 관한 연구

최용경,정태화,최명자,김재화,최인성,김용호,송은영,이희구 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1996 Journal of biomedical laboratory sciences Vol.2 No.1

환자의 복수와 늑막액으로부터 p-diazonium phenylphosphorylcholine(DPPC) coupled Separose-4B affinity chromatography와 hydroxylapatite chromatography를 실시하여 C-reactive protein (CRP)를 분리, 정제하였다. 정제된 CRP를 토끼에게 면역화하여 항혈청을 얻고 affinity chromatography를 하여 면역항체(IgG)를 분리하였다. 분리된 면역항체를 미립자에 감작시킨 후 미립자 응집반응에 의하여 3분내에 CRP를 측정할 수 있는 간이 면역측정법을 개발하였다. 본 연구에서 개발된 CRP측정법의 검출범위는 0.5∼20㎎/㎗이며, 임상 시험 결과 0.7∼2.9㎎/㎗에서는 강한 응집반응을, 5.0∼13.2㎎/㎗에서는 약한 응집반응을 보였고 28㎎/㎗이상에서는 항원 과잉으로 인한(zone of Ag excess phenomenon) 위음성을 나타냈다. 74명의 환자 혈청을 대상으로 CRP의 농도를 조사한 결과 평균치는 3.8㎎/㎗이었으며 대부분의 환자에서는 10㎎/㎗ 이하의 농도로 존재하였다. 그러므로 1차판정시 음성을 나타낸 시료라도 혈청을 5∼10배정도 희석하여 재분석한다면 오차없이 CRP를 검출할 수 있었다. 환자 혈청을 검체로 하여 본 연구에서 개발한 면역측정법과 현재 수입 시판중인 프랑스의 B사 제품과 일본의 I사 제품을 비교한 결과 좋은 상관관계를 보였다. 이와 같은 평가 분석을 통하여 볼 때 본 연구에서 개발한 간이 면역측정법은 사용이 비교적 간편하며 신빙성이 있어 CRP를 스크리닝 하는데 효과적임을 알 수 있었다. The C-reactive protein(CRP) from ascitic and pleural fluid was purified using calcium dependent affinity chromatography of CNBr activated Sepharose-4B covalently coupled to p-diazonium phenylphosphorylcholine(DPPC) and hydroxylapitite chromatography. Polyclonal antibody was prepared from rabbit by immunizing the purified CRP. Specific immunoglobulin G was isolated using affinity chromatography and coupled to microparticles. A sensitive microparticle-based immunoassay was developed to measure CRP within 3 mins. The detection range was between 0.5㎎/㎗ and 20㎎/㎗ in serum, showing strong response in the range of 0.7∼2.9㎎/㎗, week response in 5.0∼13.2㎎/㎗ and zone phenomenon over 28㎎/㎗. The average value of CRP in 74 samples was 3.8㎎/㎗ and most of the values were lower than 10㎎/㎗. The CRP values of serum samples were determined by our microparticle-based immunoassay, and were compared with those obtained using the other commercial products(B Co., France and I Co., Japan). Good correlations were shown between the values obtained by our developed microparticle-based immunoassay system and those by other commercial products. All performance characteristics evaluated make our developed microparticles-based immunoassay suitable for a simple, rapid, and reliable screening of CRP in serum.

機械技術이 造形藝術에 미친 影響

鄭仁和 진주교육대학교 1983 論文集 Vol.27 No.1

國民學校 美術科 學習指導의 問題點과 改善方向에 關한 硏究

鄭仁和 진주교육대학교 1986 論文集 Vol.30 No.1

합성 펩티드를 이용한 Interleukin-2수용체 β-Chain에 대한 단일클론항체의 제작

이희구,이홍수,이인애,최용경,최인성,정태화,김길현 이화여자대학교 생명과학연구소 1995 생명과학연구논문집 Vol.6 No.-

Hybridoma producing monoclonal antibodies reactive with the β-chain of interleukin-2 receptor (IL-2R) were generated from mice immunized with a synthetic peptide of 19 amino acids. The synthetic peptide constitutes strongly hydrophilic region of extracellular domain of the IL-2R molecules(residues 32-50) as revealed by hydrophathy plot analysis. Hybridomas were screened by ELISA using Hut102 cells and the synthetic peptide as antigenic moiety, and by miasuring the binding activity of antivodies to the Hut102 cell surface that was monitored by a flow cytometer. The antibodies were further screened by their inhibitory effect against the interaction between IL-2R and interleukin-2(IL-2). Antibodies were further characterized by immunoprecipitation followed by autoradiography to reveal that the antibodies recongnize the 75 KD protein molecules expressed on the Hut102 cell surface and human peripheral blood mononuclear cells(PBMC) shich confirms the antigenic moiety to be β-chain of IL-2 recepter molecules.

합성 펩티드를 이용한 Interleukin-2수용체 β-Chain에 대한 단일클론항체의 제작

이희구,이홍수,이인애,최용경,최인성,정태화,김길현 梨花女子大學校 藥學硏究所 1995 藥學硏究論文集 Vol.- No.5

Hybridoma producing monoclonal antibodies reactive with the ß-chain of interleukin-2 receptor (IL-2R) were generated from mice immunized with a synthetic pepride of 19 amino acids. The synthetic peptide constitutes strongly hydrophilic region of extracellular domain of the IL-2R molecules(residues 32-50) as revealed by hydrophathy plot analysis. Hybridomas were screened by ELISA using Hut102 cells and the synthetic peptide as antigenic moiety, and by measuring the binding activity of antibodies to the Hut102 cell surface that was monitored by a flow cytometer. The antibodies were further screened by their inhibitory effect against the interaction between IL-2R and interleukin-2(IL-2). Antibodies were further characterized by immunoprecipitation followed by autoradiography to reveal that the antibodies recongnize the 75 KD protein molecules expressed on the Hut 102 cell surface and human peripheral blood mononuclear cells(PBMC) which confirms the antigenic moiety to be ß-chain of IL-2 recepter molecules.

B형 간염 바이러스 단백질에 있어서 HLA-A2에 의해 표현되는 Epitope 펩타이드 들의 분석

이희구,임종석,김승목,이기영,김희수,김승호,권태종,최인성,정태화,김길현 梨花女子大學校 藥學硏究所 1995 藥學硏究論文集 Vol.- No.5

The cytotoxic T lymphocyte (CTL) are an important component in host defense mechanism against viral infection. They can recongnize virus-derived peptides presented by the Class I MHC molecule at the cell surface of the infected cells. On searching for effective CTL epitopes of hepatitis B virus(HBV), we synthesized a distinct set of 9-10 mer peptide containing amino acid sequence of hepatitis B virus surface protein that are selected on the basis of a computer modeling and the previously described HLA-A2 specific motifs.Binding assay of the synthetic peptides to HLA-A2 molecules using human antigen processing defectant T2 cells showed that 3 out of 4 synthetic peptides enhanced the expression of HLA-A2 molecule on T2 cell surface.Two anchor positions, namely P2 and P9(or P10) appeared to play a decisive role for binding.Structural. characteristics of the peptides addressed by molecular dynamics simulation was analysed and compared.These peptides also partially triggered CTL isolated from human peripheral blood mononuclear cells of HBV positive patients, and the response was peptide-spcific.These results showed that negatively-charged amino acid residue at P2 hampered binding affinity of the peptides to HLA-A2 molecules, and that binding affinity of the peptides are not always reflected by their immunogenicity among natural T cell repertoire.

B형 간염 바이러스 단백질에 있어서 HLA-A2에 의해 표현되는 Epitope 펩타이드 들의 분석

이희구,임종석,김승목,이기영,김희수,김승호,권태종,최인성,정태화,김길현 이화여자대학교 생명과학연구소 1995 생명과학연구논문집 Vol.6 No.-

The cytotoxic T lymphocyte(CTL) are an important component in host defense mechanism against viral infection. They can recongnize virus-derived peptides presented by the ClassⅠ MHC molecule at the cell surface of the infected cells. On searching for effective CTL epitopes of hepatitis B virus(HBV), we synthesized a distinct set of 9-10 mer peptide containing amino acid sequence of hepatitis B virus surface proteion that are selected on the basis of a computer modeling and the previously described HLA-A2 specific motifs. Binding assay of the synthetic peptides to HLA-A2 molecules using human antigen processing defectantn T2 cells showed what 3 out of 4 synthetic peptides enhaced the expression of HLA-A2 molemule on T2 cell surface. Two anchor positions, namely P2 and P9(or P10) appeared to play a decisive role for binding. Structural chacteristics of the peptides addressed by molecular dynamics simulation was analysed and compared. These peptides also parially triggerd CTL isolatied frmo human peripheral blood mononuclear cells of HBV positive patients, and the response was peptide-specific. These results showed that negatively-charged amino acid residue at P2 hampered binding affinity of the peptides to HLA-A2 molecules, and that binding affinity of the peptides are not always reflected by thier immunogenicity among natural T cell repertoire.