Aminoglycoside Antibiotic Modifying Enzymes in Aminoglycoside Producing Streptomycetes and Micromonospora spp.

구양모,최석례,김공환,임번삼,이새배,Goo, Yang-Mo,Choi, Seok-Rye,Kim, Kong-Hwan,Lim, Bun-Sam,Lee, Sae-Bae 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.4

Streptomycin을 생성하는 7종의 S. griseus와 1종의 S. galbus의 세포 추출물은 streptomycin을 phosphorylation하여 비활성화 하였다. Phosphorylation에 의한 streptomycin의 비활성화는 모든 streptomycin 생성균주에 공통적으로 존재하는 것으로 생각된다. Neomycin과 paromomycin을 생성하는 S. fradiaea와 S. rimosus forma paromomycinus의 세포추출물은 phosphorylation과 acetylation에 의하여 neomycin을 비활성화하였다. Kanamycin을 생성하는 S. kanamyceticus IFO 13414의 세포추출물도 neomycin을 phosphoryltion 또는 acetylabon시켜 비활성화하나 이 추출물에는 kanamycin에 대한 phosphorylation 효소는 존재하나 acetylation 효소는 존재하지 않은 것으로 확인되었다. 다른 kanamycin 생성균주인 S. kaηamyceticus NRRL 2535는 neomycin과 kanamycin에 강한 내성을 보였으나 이들 항생물질은 acetylation 시키거나 phosphorylation 시키는 효소는 확인되지 않았다. Gentamicin 또는 slsomicin을 생성하는 Micromonospora spp. 들은 gentamicin, kanamycin 그리고 tobramycin에 강한 내성을 보였으나 어떤 종류의 aminoglycoside 항생물질도 phosphorylaton 시키거나 acetylation 시키는 효소는 확인되지 않았다. Aminoglycoside 항생물질을 생성하는 Streptomycetes 들은 자신들이 생성하는 항생물질 뿐만 아니라 유사한 구조를 갖는 다른 항생물질에 대하여서도 특이한 내성메카니즘을 소유하고 있는 것으로 확인되었다. The cell free extracts of streptomycin producers, 7 strains of S. griseus and 1 strain of S. galbus inactivate streptomycin by phosphorylation, and the resistant mechanism against streptomycin by phosphorylation seems to be popular in streptomycin producing organisms. The cell free extracts of neomycin and paromomycin producers, S. fradiae and S. rimosus forma paromomycinus inactivate neomycin not only by phosphorylation, but also by acetylation. The cell free extract of a kanamycin producer. S. kanamyceticus (IFO 13414) inactivates neomycin by phosphorylation and/or by acetylation. However it does not inactivate kanamycin by phosphorylation but does by acetylation. The cell free extracts of the other kanamycin producer. S. kanamyceticus (NRRL 2535) does not show any inactivation of neomycin as well as kanamycin by phosphorylation or by acetylation, although it shows strong resisance against the antibiotics. The cell free extracts of Micromonospora spp.. producers of gentamicin or sisomicin which show resistance against gentamicin, kanamycin and tobramycin do not possess any phosphorylating or acetylating enzymes for the aminoglycoside antibiotics employed in this study. The Streptomycetes producing aminoglycoside antibiotics seem to have characteristic resistance mechanisms against the antibiotics produced by themselves and against other similar antibiotics.

Aminoglycoside 항생물질을 생성하는 Streptomycetes 와 Micromonospora spp 들의 Aminoglycoside 항생물질 변형효소에 관한 연구

구양모,최석례,김공환,임번삼,이새배 ( Yang Mo Goo,Seok Rye Choi,Kong Hwan Kim,Bun Sam Lim,Sae Bae Lee ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.4

The cell free extracts of streptomycin producers, 7 strains of S. griseus and 1 strain of S. galbus inactivate streptomycin by phosphorylation, and the resistant mechanism against streptomycin by phosphorylation seems to be popular in streptomycin producing organisms. The cell free extracts of neomycin and paromomycin producers, S. fradiae and S. rimosus forma paromomycinus inactivate neomycin not only by phosphorylation, but also by acetylation. The cell free extract of a kanamycin producer. S. kanamyceticus (IFO 13414) inactivates neomycin by phosphorylation and/or by acetylation. However it does not inactivate kanamycin by phosphorylation but does by acetylation. The cell free extracts of the other kanamycin producer, S. kanamyceticus (NRRL 2535) does not show any inactivation of neomycin as well as kanamycin by phosphorylation or by acetylation, although it shows strong resisance against the antibiotics. The cell free extracts of Micromonospora spp.. producers of gentamicin or sisomicin which show resistance against gentamicin, kanamycin and tobramycin do not possess any phosphorylating or acetylating enzymes for the aminoglycoside antibiotics employed in this study. The Streptomycetes producing aminoglycoside antibiotics seem to have characteristic resistance mechanisms against the antibiotics produced by themselves and against other similar antibiotics.

국산 항 CEA 항체의 I - 131 , Tc - 99m 표지법 확립 및 면역학적 특성 분석

고창순(Chang Soon Koh),이명철(Myung Chul Lee),정준기(June Key Chung),이동수(Dong Soo Lee),홍미경(Mee Kyoung Hong),최석례(Seok Rye Choi),서일택(Il Taek Seo),정홍근(Hong Keun Chung),정준호(Jun Ho Chung) 대한핵의학회 1992 핵의학 분자영상 Vol.26 No.2

N/A Caneer cells have several tumor-associated antigens on the cell surfaces, and antibodies against these antigens have been developed by many investigators. Radiolabeled antibodies have been used as new methods to diagnose and treat malignant tumors. Especially anti- carcinoembryonic antigen (CEA) is the most popular antibody for these purposes. In this investigation, we tried to label 131I and Tc-99m to anti-CEA monoclonal antibodies which were developed in the Seoul National University College of Medicine. We found CEA-79 and CEA-92 antibodies had the better immunological characteristics among 8 anti-CEA monoclonal antibodies. And radioiodination of CEA-79 could be performed by chloramine-T method, while radioiodination of CEA-92 by iodogen method. To label these antibodies with Tc-99m, we used pretargeting transchelation as direct labeling method. At first, Tc-99m was bound to glucaric acid, and monoclonal antibody was reduced by β-mercaptoethanol. When these were incubated together, Tc -99m bound to glucarate was switched to monoclonal antibody because of higher affinity. We established conditions of several steps in this method. Anti-CEA monoclonal antibodies labeled with 131I and Tc-99m are expected to be used valuably in the detection and treatment of malignant tumors.

항 NCA - 95 단일클론항체의 99mTc 표지 키트 제조 및 특성 연구

고창순(Chang Soon Koh),이명철(Myung Chul Lee),정준기(June Key Chung),이동수(Dong Soo Lee),정재민(Jae Min Jeong),김채균(Chae Kyun Kim),홍미경(Mee Kyoung Hong),최석례(Seok Rye Choi),이용진(Yong Jin Lee) 대한핵의학회 1996 핵의학 분자영상 Vol.30 No.4

N/A The previous monoclonal antibody labeling method for bone marrow immunoscintigrapy was complicated and laborious for clinical application. Also it showed a relatively low labeling efficiency. To improve this procedure, we compared several direct labeling methods of Tc-99m. 1) The labeling efficiency in the method using gluconate as a transchelator was low (40-70%), but immunoscintigraphy using this radiotracer produced a clear image. 2) To improve labeling efficiency, β-mercaptoethanol was removed after reduction. The labeling efficiency was improved up to 70-80%, but the radioactivity of the blood pool was high. 3) The higest labeling efficiency (〉90%) and best quality images could be obtained by using MDP as a transchelating agent. It did not require additional procedures for separation of labeled antibodies. The immunoreactivity of this antibody was 60%. Residual MDP which can be taken up by the bone could be removed by PD-10 column. The reduced antibodies were stable with a high labeling efficiency (〉90%) for up to 47 days by deep freezing. We concluded that the improved procedure for Tc-99m labeling of anti-NCA-95 monoclonal antibody using MDP as a transchelating agent will be a simple and useful method for clinical application.

실험적 뇌허혈증 모델에서 허혈 조직의 99mTc - glucarate 섭취

고창순(Chang Soon Koh),이명철(Myung Chul Lee),정준기(June Key Chung),이동수(Dong Soo Lee),김영주(Young Ju Kim),정재민(Jae Min Jeong),김채균(Chae Kyun Kim),최석례(Seok Rye Choi),마응천(Woong Chun Mar) 대한핵의학회 1996 핵의학 분자영상 Vol.30 No.4

N/A To detect ischemic tissue in experimental model of cerebral ischemia made by middle cerebral artery(MCA)-occlusion, we acquired triple image of Tc-99m-glucarate, [18F]fluoro-deoxyglucose (FDG), and 2,3,5-triphenyltetrazolium (TTC) staining. We made cerebral infarction either with reperfusion (after occlusion of 2 hours) or without reperfusion in 10 Sprague-Dawley rats by inserting thread to MCA through internal carotid artery. After 22 hours, we injected 740 MBq of Tc-99m-glucarate and 55.5 MBq of [18F]FDG through tail vein. Each 1 mm slice of rat brains was frozen and exposed to imaging plate for 20 minutes in freezer to get an [18F]FDG image. After 20 hours enough to fade radioactivity of [18F]FDG, the slices were again imaged by BAS1500 for Tc-99m-glucarate uptake·Finally, these braim tissues were stained with TTC. semi-quantitative visual analysis was done by grading 0 to 3 points according to the degree of uptakes(Tc-99m-glucarate) and decreased uptakes([18F]FDG and TTC). Ten rats survived with neurologic symptoms. TTC staining confirmed the development of infarction. The size of the infarction was relatively larger in the group without reperfusion. [18F]FDG images were similar to TTC-stained images. However, we found regions with intermediate uptake which were not stained with TTC. We found regions with intermediate [18F]FDG uptake where TTC staining was normal. Tc-99m-glucarate uptake was found only in TTC non-stained region. In the TTC stained regions, there were no uptake of Tc-99m-glucarate. We could not find clear relation between Tc-99m- glucarate uptake with [18F]FDG uptake. This was partly because percent uptake of Tc-99m- glucarate was so small (less than 1 percent of injected dose) and because there were quite heterogeneity of patterns of [18F]FDG uptake and TTC. With these findings, we could conclude that Tc-99m-glucarate were taken up only in part of ischemic tissues which were proven to be nonviable. The establishment of MCA-occluded rat model with or without reperfusion and triple imaging for Tc-99m, 18F and TTC helped the characterzation of Tc-99m-glucarate uptakes. Further work is needed to clarify the meaning of diversities of [18F]FDG and TTC and their relation with Tc-99m-glucarate.

뇌혈류 평가용 99mTc - HMPAO 합성 및 분포에 관한 연구

고창순(Chang Soon Koh),이명철(Myung Chul Lee),정준기(June Key Chung),조보연(Bo Youn Cho),이범우(Bum Woo Lee),정재민(Jae Min Jeong),염미경(Mi Kyung Yeom),홍미경(Mee Kyoung Hong),최석례(Seok Rye Choi) 대한핵의학회 1990 핵의학 분자영상 Vol.24 No.2

N/A HMPAO was synthesized by two step reaction. d, 1-HMPAO was separated from the racemic product by fractional recrystalization in ethylacetate, and the chemical structure and purity was confirmed by proton NMR spectroscopy. The synthesized d, 1-HMPAO was labeled with Tc- 99m and studied the biodistribution in mice. From the results we could find that liver uptake of synthesized Tc-99m d, 1-HMPAO was higher than that of Amersham kit, but no conspicuous difference was found in brain and other tissues (blood, lung, stomach, intestine, muscle, spleen and kidney).