間接螢光抗體法 및 免疫黃金標識法을 이용한 肝吸蟲組織內 抗原性部位에 관한 硏究

成大林,金洙鎭,嚴基善,林漢鍾 고려대학교 의과대학 1988 고려대 의대 잡지 Vol.25 No.3

Indirect fluorescent antibody test and immunogold labeling method were accomplished to observe the antigenic localization in the tissues of adult Clonorchis sinesis. Immunoglobulins (IgG) for the reaction were obtained from white rats and rabbits, at eight weeks after experimental infection with 50 and 500 metacercariae of C. sinensis, respectively. Frozen sectioned adult worm antigen (8μm) and FITC-conjugated goat anti-rat, rabbit IgG were used for indirect fluorescent antibody test. On the other hand, sectioned tissue antigen embedded in Lowcryl HM 20 medium and secondary antibody protein A gold complex (particle size:12 nm) were used for immunogold labeling method. In the results, fluorescence or gold particle were not observed on the tissue antigens of the worm when reacted with the noninfected control IgG. However, with the infected animal groups IgG, fluorescence or gold particles were observed on the interstitial matrix (body fluid) between the various organs, such as the tegumental cells, parenchyma, testes, receptaculum seminis. And the cecal epithelium including cecal contents and epithelium of excretory bladder also showed strong reaction. Therefore, antigenic materials which stimulate the IgG antibodies against the host were thought to be fluid substances and digestive enzymes or excretory materials which make up the interstitial matrix in the tissues of the worm.

Chinese hamster 난소세포에서 수용성 니켈에 의해 특징적으로 발현되는 유전인자들의 빠른 분석에 관한 연구

성근제,이상한,우기민,송호연,조만희 순천향의학연구소 2001 Journal of Soonchunhyang Medical Science Vol.7 No.2

니켈에 의한 암 발생기전 및 세포 독성기전을 이해하고 니켈에 의해 발현에 변화를 보이는 유전자들을 검출하기 위하여 니켈이 포함된 배지에서 배양된 Chinese hamster난소 세포부터 mRNA differential display 분석을 시행하였다. 본 연구에서는 동위원소나 DNA염기서열 분석용 젤을 사용하는 기존의 방법 대신에 훨씬 빠르고, 다루기 쉽고 안전성을 지닌 변형된 기법을 개발하여 적용하였다. 니켈에 의해 발현에 변화를 보이는 cDNA fragments들을 10% polyacrylamide mini젤상에서 분리 및 클로닝한 다음, DNA염기 서열 분석과 GenBank Blast search 프로그램을 이용하여 상동성 조사를 하였다. Reverse Northern blot을 이용하여 발현에 변화가 있는 cDNA들을 재확인한 결과, 검사된 19 cDNAs 들중 총 9종에서 발현의 변화를 보였으며, 이 중 3종은 니켈에 반응하여 발현에 증가를 보였고, 반면에 6종은 발현이 감소하였다.

Timoshenko 보 이론을 이용한 층상균열이 있는 보의 진동해석 및 비파괴적 평가

韓秉基,李晟熙,柳澤仁 弘益大學校 科學技術硏究所 1994 科學技術硏究論文集 Vol.5 No.-

A study of the natural vibrations of Timoshenko beam with a split is presented. An analytic model, based on the transverse and longitudinal vibration of beams is employed to determine the natural frequencies. The variations of natural frequencies with the changes of size and location of laminar tear are plotted for several thickness ratio. A specific example are given to show the effects of rotary inertia, and shear deformation on the natural frequencies of the beam with a split. A new skill for the presentation of relationship between the size and location of laminar tear and natural frequencies is presented.

사립체 DNA 뉴클레오타이드 3243 A→G 돌연변이 [MTTL1^*MELAS3243G]에 의한 MELAS 증후군

박기형,김승현,이영주,김희태,김주한,김명호,박성섭 대한신경과학회 1998 대한신경과학회지 Vol.16 No.4

보의 층상균열의 위치 및 크기가 진동특성에 미치는 영향

韓秉基,柳澤仁,李晟熙 弘益大學校 科學技術硏究所 1993 科學技術硏究論文集 Vol.3 No.-

This study aims at finding a effect of various position and length of laminar tear on vibrational characteristics on beam including shear deformation. General solution for nonlaminar tear and laminar tear regions are first established. Recurrence equations relating integration constants for adjacent interior regions are established by satisfying continuity conditions at junctions of interior region. The frequency determinant is then obtained by satisfying contiuity conditions at junction between end region and interior regions immediately next to the end region. Numerical results are compared for experimental and references results.

사람 뇌척수액중 Transthyretin의 항산화작용

양성렬,최기오,박종근,류문희,홍석노,김수한,안봉환,이제혁,이민화 대한신경외과학회 1994 Journal of Korean neurosurgical society Vol.23 No.4

Protective effects of human cerebrospinal fluid antioxidants against enzyme inactivation caused by metal-catalyzed oxidation systems were investigated. When purified glutamine synthetase(GS) was incubated with human cerebrospinal fluid(CSF), the enzyme was progressively inactivated. Catalase and EDTA could inhibit the enzyme inactivation by 50-80%. Small-molecular(Mr< -10,000) fraction of CSF inactivated the exogenous GS, but large-molecular(Mr> -10,000) fraction did not. The GS inactivation by the small-molecular fraction was also markedly inhibited by catalase and EDTA. These results suggested that metal-catalyzed oxidation is involved in the GS inactivation by the small-molecular fraction of CSF. Dithiothreitol(DTT) was shown to inhibit almost completely the oxidative inactivation of GS by CSF. However, DTT inhibited only partially the oxidative inactivation of GS caused by small-molecular fraction of CSF. When large-molecular fraction of CSF was separated by anion-exchange HPLC chromatogrpahy, there was a peak of antioxidant activity inhibiting the small-molecular fraction-induced GS inactivation in the presence of DTT. The antioxidant activity was neutralized by monoclonal antibodies to transthyretin. Purified transthyretin was found to efficiently inhibit ascorbate/Cu^(2+)-induced GS inactivation in the presence of DTT. Uric acid and glucose did not show any protective effect on the GS inactivation in the same condition. The above results suggest that metal-catalyzed oxidation occurs normally in human CSF, and that transthyretin may play an important role as a CSF antioxidant in protecting proteins from metal-catalyzed oxidation.

인간의 혈장아포지단백 A-I에 특이성을 지닌 단일클론항체 A-I30의 light chain의 가변부위를 coding하는 cDNA의 클로님

이상한,우기민,조만희,장예진,김창세,김정경 순천향의학연구소 1997 Journal of Soonchunhyang Medical Science Vol.3 No.2

I prepared a monoclonal antibody (mAb) A-l30 to HDL apolipoprotein A-l with the ultimate goals of expressing the valuable immunodiagnostic single-chain Fv (scFv) in Escherichia coli. The binding specificity of mAb A-l30 was determined by Western blot analysis. From the hybridoma cell line secreting mAb A-l30, poly(A)+ RNA was prepared and used as a template for cDNA synthesis and cloning. The nucleotide sequence analyses revealed that the variable regions of the heavy and light chains were members of mouse heavy chain subgroup ⅡA and κ light chain subgroup Ⅱ, respectively. Comparison of the nucleotide sequences with mouse immunoglobulin genes listed in the GenBank data base showed that the cDNAs have not been previously reported.

단백질 및 1가 양이온을 통한 proteasome의 활성조절에 관한 연구

우기민,이상한,조만희 순천향의학연구소 2004 Journal of Soonchunhyang Medical Science Vol.10 No.1

Proteasome은 일반적으로 세포내 환경과 매우 상이한 조건하에서 연구된다 (실험용 완충액에는 1기가 이온들이나 여러 단백질들이 결여되어 있다). 특이하게도 본 연구결과 낮은 농도의 Na^(+)와 K^(+)는 20S proteasome의 단백질 분해 활성을 현저히 저하시키지만 26S proteasome의 단백질 분해 활성에는 크게 영향을 미치지 못하였다. Na^(+)와 K^(+)는 20S proteasome의 chymotrypsin 유사 분해활성을 1-2 mM의 Ki 값으로 매우 민감하게 억제하였다. 이에 비해 기타 다른 1가 양이온 (Li^(+), Rb^(+), 또는 Cs^(+))은 미미한 억제효과를 나타내고, 2가 양이온 (Mg^(2+))은 20S proteasome의 chymotrypsin 유사 분해활성을 오히려 활성화시켰다. 이러한 이온들은 trypsin 유사 활성과 post-acidic 활성에도 유사한 억제효과를 보였다. 또한, Na^(+)와 K^(+)는 20S proteasome에 의한 casein, oxidized RNase A, a-lactalbumin 같은 단백질들의 분해를 강력하게 억제시키지만, 26S proteasome에 의한 ATP-의존적 단백질 분해 활성에는 영향을 미치지 못한다. 일반적으로 세포내 단백질농도는 매우 높은 것으로 알려져 있는 바, proteasome의 분해활성에 미치는 단백질의 영향을 조사한 결과, 무작위의 다양한 단백질들(0.1㎎/㎖)도 1가 양이온(Na^(+)와 K^(+))의 경우와 같이 20S proteasome의 펩타이드 분해활성을 억제하였지만 26S proteasome의 활성에는 아무런 영향을 미치지 않았다. 이러한 억제작용은 경쟁적이지 않다. BSA나 ovalbumin 같은 비가수분해 단백질들이 20S proteasome의 펩타이드 분해활성을 강력히 억제한였으므로 이러한 억제현상이 비경쟁적이라는 것을 알 수 있다. 비록 Na^(+), K^(+)와 단백질들이 20S proteasome을 비활성화 형태로 유지시키지만, 0.02% SDS에 의한 20S proteasome의 활성증가를 막거나 억제시키지는 못하였다. 이 결과로 고농도의 이온과 단백질이 존재하는 세포내 조건에서는, 독립적인 20S proteasome은 잠재적으로 비활성화된 형태로 존재하며, 19S ATPase 복합체나 11S 같은 proteasome 활성복합체가 결합한 후에 비로소 26S proteasome의 활성화된 형태로 전환되는 기작을 갖는 것으로 여겨진다. 또한, Na^(+)나 K^(+), BSA나 ovalbuim의 첨가는 비활성화 형태의 20S proteasome을 이용한 많은 연구를 보다 용이하게 할 것으로 기대된다.

Thermoplasma acidophilum의 20S Proteasome에 의한 ATPase-활성적 단백질분해에 관한 연구

우기민,장예진,조만희,김창세,김완종,조성호,이상한 순천향의학연구소 1999 Journal of Soonchunhyang Medical Science Vol.5 No.1

The eukaryotic 26S proteasome is an ATP/ubiquitin-dependent proteolytic complex consisting of the 20S core particle and 19S ATPase complex. However, because of its complexity and unstable properties, this study was carried out to present more simple and stable model for the ATP-activated proteolytic complex in prokaryotes which can take the place of the eukaryotic 26S proteasome. For this purpose, recombinant Thermoplasma 20S proteasome (T20S) and Methanococcus MS4, a sequence homolog of one ATPase subunit in the 19S ATPase complex, were successfully isolated from Escherichia coli (E. coli). The α and β subunits of T20S expressed in E. coli could assemble for themselves, and showed the peptide-hydrolyzing activity. Whereas both T20S and R20S (the 20S complex from rabbit skeletal muscle) had the highest peptidase activity against Suc-LLVY-AMC, a good substrate for chymotrypsin-like peptidase activity, the specific activity of T20S was slightly lower than that of R20S. In addition, several reagents such as KCI, SDS, and ovalbumin were shown to have different effects on the peptidase activities between T20S and R20S. When the ATPase activity of the purified MS4 were assayed , the Km for ATP was about 0.5 mM, and casein could stimulate the activity more than 2 fold without the change in Km. This result implicates that protein-activated ATPase may induce the conformational change of casein, and therefore suggests that MS4 ATPase may activate the proteolytic activity of the 20S proteasome via accelerating the recognition and translocation of the protein substrates.

단층촬영 각도의 변화가 하악과두의 골 증식성 병소의 인식에 미치는 영향

김기덕,한상선 大韓口腔顎顔面 放射線學會 1999 Imaging Science in Dentistry Vol.29 No.1

Purpose: To find out the effects that different tomographic angles have on the osteophytic lesion detectability of condyle head by comparison the individualized lateral tomographic image with the various tomographic angled images using SCANORA . Materials & Methods: This study is performed to simulate osteophytic lesions by a series of dentin chips placed at six locations on condyle head. The control angle is 15°and from this angle, tomographic angle were varied with -10°, +10°+20°. All the images with each sized dentin chip were scored by three dental radiologists with the use of confidence levels for presence or absence of the lesion, each examiner viewed one of the images twice. A rating scale from 0 to 2 (0, lesion definitely not present; 1, uncertain if lesion is present; 2, lesion definitely present). Responses were assessed by Tukey's multiple comparison method and kappa value. Results: 1. The lesion size of 0.3mm could not be detected in all the tomographic angles. As the size of the lesion increased the average value of lesion detectability also increased. 2. In the lesion sizes of 0.7mm there was statistically significant difference between the 15°control angle and the altered tomographic angles (p<0.05). In 1.0mm lesion there was no significant difference in the ±10°altered angles (p<0.05), but there was significant difference in the altered angle(p<0.05). In the lesion sizes of 0.3mm and 2.0mm there was no significant difference between the 15°control angle and all the altered angle(p>0.05). 3. In the anteromedial, anterosuperior, anterolateral area there was no significant difference between the 15°control angle and the ±10°altered angle(p>0.05), but in the comparison with the +20°alterd angle there was significant difference(p>0.05). Conclusion: When imaging the lateral tomography of the temporomandibular joint used by SCANORA, it can be considered that in the osteophytic lesion size of 2mm and above, the tomographic angle difference within +20°to the horizontal angle of the condyle, has little effect on the lesion detectability. And in the lesion size of 1mm, the altered angle within ±10°also has little effect on the lesion detectability.