관상 동맥 질환에서 아포 E 지단백 유전자 다형성과 혈청 지질치와의 관계

곽선영,김성구,정호석,이유경,이광희,김철현,최태명,현민수,권영주 순천향의학연구소;Soonchunhyang Medical Research Institute 2000 Journal of Soonchunhyang Medical Science Vol.6 No.1

Background and aims: The Apolipoprotein E is a ligand of both the protein component LDL receptor as well as the apo E LDL-Receptor related protein (LRP). It modulates the receptor binding of lipoproteins, with the apolipoprotein E found on cell surfaces as its component, thus serving an important role in the lipid metabolism by carrying out the intracellular transport of cholesterol in lipoproteins. The gene for apolipoprotein E is the product of three common genotypes as well as many more rare alleles. The common genotypes are ε2, ε3, and ε4, and are expressed in the three phenotype isoforms of E2, E3, and E4. In the event that E4 is the main component, a rise in the cholesterol level, as the result of down-regulation of the LDL receptor, is observed. Therefore, those samples with E4 genotypes are known to be in much higher risk of coronary artery disease than those with ε3/ε3, while those with ε2 are in low risk (with the exception of hypertiglyceremai Ⅲ). The aim of this study is to analyze in patients with ischemic heart disease the role of aplipoprotein E alleles in order to seek its correlation with coronary artery disease, as well as to seek whether the polymorphism of apo E produces any differences in the severity of coronary artery disease according to plasma lipid levels. Methods: The subjects for study were 273 patients admitted to the Internal Cardiology Division of the Soonchunhyang University Hospital form December 1998 to February 1999. The subjects were divided into the two groups of which one was ischemic heart disease (IHD) experiment group totaling 105 (avg.60.1 years of age, male/female ratio = 69/36) and the control group totaling 168 (avg. 59.7 years of age, male/female = 73:95). The coronary angiogram was given to 127 subjects, and of this total, 94 have developed significant stenosis in the coronary artery. The stages of the analyzing of the apo E phenotype was first, the separation of DNA from the blood samples, subjecting it to the PCR from with 228 base pairs of expanded products were obtained. The band was determined by means of the reverse hybridization principle on the nitrocellulose strip. Results: From the 105 patients the distributions of apo E phenotypes were as follows: ε3/2(5.7%), ε4/2(1.9%), ε3/3(70%), ε4/3(20%), ε4/4(1.9%). The relative frequencies of each allele are as follow: ε2 (0.038), ε3 (0.833), ε4 (0.128). The results show as follows: ⅰ) The IHD experiment group to have a higher occurrence of ε4/3 phenotypes as well as ε4 alleles than the control group. ⅱ) Both the control group and IHD group showed the largest distribution of ε3/3 for phenotypes, and ε4 for alleles. ⅲ) The IHD group showed less ε2/3 phenotypes as well as significantly less allele frequency of ε3 in comparison to the control group. ⅳ) the IHD group showed a much lower level of HDL in comparison to the control group, while the LDL was significantly higher; samples including the apo ε2 showed a significantly higher level of HDL than those without. Among the control group, samples including apo ε2 showed a significantly higher level of TG (triglyceride) than samples without. No significant difference was found between the experiment apo ε4 sample and the control plasma lipid sample. ⅴ) No significant correlation was found between an apo E polynorphism and the number of involved arteries of a coronary angiogram. Conclusion: Between the experiment IHD group and control group were found differences in the frequency of alleles. The polymorphism of apo E alleles may contribute as a risk factor to the development of heart disease by involving itself in the metabolism and modulation of plasma lipids.

무 추출물이 식품관련 미생물의 증식에 미치는 영향

곽희진,계수경,곽희선,이경혜 동아시아식생활학회 2000 동아시아식생활학회지 Vol.10 No.4

To understand the effect of radish on growth of food born microorganisms, mashedflesh radishes were extracted by using acetone and distilled water. Their effect was assayed by measuring the optical density of cultural broth of food born microorganisms, In the experiment, seven strains of food born bacteria and one strain of yeast were used as the test organism. Acetone extract inhibited growth of the cells of L. plantarum, L. sake and Danmuji film yeast. Growth of the film yeast was drastically inhibited in the concomitant presence of 0.03% extract, while other microbes such as L. faecalis. P. pentosaceus, B. subtilis and E. coil grew by succeeding cultivation for 4 to 8 hours after addition of the extract. Water extract, on contrast to acetone extract, at the concentrations of 0.1~1.5% stimulated the growth of lactic acid bacteria, Culture of L. faecalis and L. sake showed an optical density higher than that of control by 40~50 times. The effect was not so apparent against E. coli, S. aureus and Danmuji film yeast.

대장균에서 재조합 Rat Guanine deaminase 유전자의 발현, 정제 및 분석

성연선,곽상준,박대성,김향원,이희영 단국대학교 1998 論文集 Vol.33 No.-

Guanine deaminase(EC, 3.5.4.3, ;Guanine aminohydrolase, GAH, GDA) catalyzes the deamination reaction of guanine to xanthine irreversibly. The cDNA encoding rat guanine deaminase had been isolated from a λZAPII rat brain expression library using antibody generated against purified rat guanine deaminase. Toe obtain recombinant GDA and analyze the property of catalytically conserved retion, here we expressed the recombinant GDA in E.coli and showed the retention of its catalytic activity similar to native rat GDA. To make the construct carrying coding region to GDA in pGEX4T prokaryotic expression vector, the region encompassing open reading frame of pBlue-GDA was PCR amplified and subcloned into pGEX4T prokaryotic expression vector with correct reading frame of fusion carrier glutathione S-transferase. After transformation to E.coli DH5a. The bacteria carrying pGEX-GDA was grwon in the condition of IPTG induction. The fusion protein GST-GDA was purified with GSH-sepharose affinity chromatography. The purified GST-GDA was subsequently digested with biotin-conjugated thrombin, and thrombin was removed by streptavidin agarose. Then sample were treated with GSH-sepharose affinity chromatography to remove the contaminant GST and GST-GDA. The purified recombinant GDA showed 70% of specific activity relative to that of purified rat GDA. The recombinant GDA showed identical molecular weight with rat GDA in SDS-PAGE.

Phosphorylation modulators에 의한 항암제 다제내성 유전자의 발현 조절

김선희,곽남희,강치덕,정병선 부산대학교 유전공학연구소 1995 분자생물학 연구보 Vol.11 No.-

암을 치료함에 있어서 항암제에 대한 다제내성(MDR)의 획득이 중대한 장애로서 지적되고 있으며, MDR 현상은 MDRI 유전자의 과잉 발현에 의한 것으로 알려져 있다. MDRI promoter 활성이 mutated ras 또는 raf-1 발현에 의하여 증강되는 현상을 좀 더 명확히 하기 위하여 암 환자의 조직검체를 사용하여 RNA slot-blot 분석법으로 MDRI 유전자의 발현과 이들 암 유전자의 발현과의 관련성을 조사하였다. 3개의 대장암 검체에서 이들 암유전자의 높은 발현과 함께 MDRI gene도 높은 발현을 나타내었다. Raf-1에 의한 MDRI promoter의 최대 활성 증강을 나타내는 DNA 배열은 이 promoter 영역내의 HSE 영역과 일치하며, 또한 이 promoter에 작용하는 regulatory clement가 has70 promoter와 매우 유사하여 heat-induced(42℃) p432-CAT 발현에 있어 protein phosphatase inhibitor의 효과를 조사하였다. Okadaic acid나 calyculin은 농도 의존적으로 heat-induced p432-CAT의 발현을 증강시켰다. 또한 p56-CAT 발현이 forskolin에 의한 증강 또는 H-87에 의한 감소를 나타내어 이러한 현상은 MDRI promoter내의 CRE 유사 배열이 기능을 나타내는 것으로 사료되어 MDRI promoter가 has70 promoter와 유사하게 조절되며, Raf-1 발현에 의한 MDRI promoter의 활성화는 MDRI promoter내에 존재하는 HSE와 관련됨을 시사하였다. Multidrug resistance(MDR) poses a serious clinical problem in chemotherapy of cancer. MDR results from overexpression of the MDRI gene which encodes a drug-efflux pump called P-glycoprotein. To confirm our observation that the expression of mutated ras or raf-1 gene increased MDRI promoter activity, it was examined whether MDRI gene expression were correlated with the expression of these oncogenes in clinical tumor samples using RNA slot-blot analysis. These oncogenes were correlated with the expression of MDRI gene in 3 colon tumor samples. Since DNA sequence exhibiting maximum activation of MDRI promoter by Raf-1 is associated with heat shock element of MDRI promoter region, and regulatory element contained in MDRI promoter is very similar to that of hsp70 promoter. The effects of okadaic acid or calyculin on the heat-induced(42℃) expression of p432-CAT were determined. Okadaic acid or calyculin A potentiated the heat-induced expression of p432-CAT in dose-dependent manner. In addition, stimulation or inhibition of p56-CAT expression by forskolin or H-87, respectively suggests functional operation of CRE element in MDRI promoter. It is suggested that the regulation of MDRI promoter activity is very similar to that of hap70 promoter.

대유행인플루엔자 대비를 위한 의료기관-바탕 탁상훈련 : 고안 및 평가

설희윤,김지량,권보란,목정하,이선희,곽임수,정진우,김정수,고옥배,조은희,김성순,신상숙,이상원 대한감염학회 2008 감염과 화학요법 Vol.40 No.2

목적 : 의료기관의 범유행 인플루엔자에 대한 효과적인 대비를 위한 훈련방법으로서 의료기관-바탕 탁상훈련 (hospital based tabletop exercise)의 유용성과 순응도를 평가하기 위해서 시행되었다. 재료 및 방법 : 탁상 훈련은 부산대학교 병원에서 시행되었으며 병원의 주요의사결정권자 및 주요부서 대표자, 실무자 42명이 지휘부, 진료부, 지원부 3그룹으로 나뉘어 훈련에 참가하였다 탁상훈련의 시나리오는 의료기관의 자체 훈련을 위해서 고안하였으며 현재 동남아에서 유행하고 있는 H5N1 인플루엔자가 국내에 처음으로 유입되어 확산되는 과정에서 의료기관에서 발생할 수 있는 상황을 3가지 모듈로 구성하였다. 훈련 평가는 훈련 전후에 각각 익명의 설문조사를 시행하였다. 결과 : 훈련에 초청된 참여자 42명 중 37명(88%)이 실제 당일 훈련에 참여하였다. 훈련에 참여한 37명의 직원 중에서 훈련 전 시행한 설문에 27명(73%)이 응답하였다. 훈련 전 정가에서는 훈련을 통해 획득하고 싶은 지식이나 기술의 우선 순위를 물었으며, 참여자들이 가장 중요하다고 생각하는 항목은 대유행인플루엔자 바이러스에 대한 지식 향상 22%, 병원 각 부서가 취해야 할 계획 및 의사소통과 관련된 지식 향상 19%, 지역사회 병원들 간의 상호협력을 향상시킬 전략 계발이 19%이었다. 그룹토의 동안에는 환자의 조기 발견 이후 대응에 대한 토론이 많았지만, 구체적인 부분이 미흡하였고, 대유행 인플루엔자에 대한 기본적인 이해가 부족하였다. 훈련에 참여한 37명의 직원 중에서 훈련 후 시행한 설문에 21명(57%)이 응답하였다. 훈련 후 평가에서 탁상훈련의 전반적인 만족도에 대한 질문에서 81%가 만족하였다고 답변을 하였고, 새로운 것을 배우는데 유용했는가 대한 질문에도 86%가 유용했다고 답변하였다. 결론 : 탁상훈련은 의료기관의 대유행인플루엔자 대비에 효과적인 훈련방법이라고 생각되며 훈련을 통해 의료기관 내의 여러 부서들이 각 의료기관의 특성에 맞는 현실적이고 실제적인 대비 계획을 수립하는데 도움을 줄 것으로 기대된다. 그러나 향후 토른 촉진, 구체적인 결과에 도달을 유도할 수 있는 질문, 끼워넣기 개발, 적절한 시간 등에 대한 연구가 필요할 것으로 생각된다. Background : To evaluate the usefulness and compliance of a hospital-based tabletop exercise in setting of pandemic influenza in hospitals. Materials and Methods : Tabletop exercise was held in Pusan National University Hospital and forty two hospital employees were invited to participate in the exercise. The scenario for hospital-based tabletop exercise was designed. It consisted of three modules, which simulated the influx and outbreak of H5N1 influenza that was epidemic in Southeast Asia. Pre-, post-exercise surveys were completed by anonymous questions. Results : Thirty-seven (88%) of 42 invited participants attended exercise. AII members of the administration group and the ancillary services group participated. But, only 77% members of the clinical services group participated. In pre-exercise survey, priorities of eight goals regarding skills and knowledge during exercise were inquired., The highest priorities pointed out by the respondents were "Increase the knowledge of pandemic influenza" (22%), "Development of strategies for optimal communication among employees within specific department" (19%) and "Development of strategies for improved coordination between facilities within the health system" (19%). Twenty-one (57%) of participants completed the post-exercise surveys. At post-exercise surveys, 81% of the participants stated that the tabletop exercise was extremely or very useful, 86% of the participants also stated that it increased their knowledge of pandemic influenza. Conclusion : Tabletop exercise is an effective modality for increasing pandemic influenza preparedness in hospitals, and this method is useful for guiding preparedness activities within the hospital environment. Further studies to determine the appropriate method of discussion, questionnaire, duration of exercise and injection are needed.

반코마이신 내성 장내구균 분리 환자에서 황색포도알균의 내성

김동욱,신선혜,김혜진,류선,이선희,장철훈,김영대,곽임수 대한감염학회 2004 감염과 화학요법 Vol.36 No.1

목적 : Vancomycin-resistant enterococci (VRE)로부터 S. aureus로의 vacomycin 내성 유전자의 전파는 오래전부터 예견되어져 왔으며 큰 걱정거리로 남아있다. 최근에 vanA 유전자를 가지고 있는 vancomycin-resistant S. aureus 2주가 미국에서 분리되었다. 이 연구의 목적은 VRE를 보균하고 있는 환자들에서 S. aureus 보균양상과 항균제 내성을 알아보는데 그 목적이 있다. 재료 및 방법 : 2001년 1월부터 2001년 12월까지 부산대학교병원에 입원하였던 환자들 중 임상검체에서 VRE가 분리된 환자 7명과 2001년 9월부터 12월까지 외과 중 환자실에 입원하였던 환자들 중 VRE 감시배양을 시행하였던 20명의 환자들을 대상으로 하였다. 환자들은 전비공, 액와부, 회음부, 직장에서 매주 면봉으로 검체를 채취하였다. 선별배지에서 검체를 배양하여 S. aureus를 분리하였고 원판 확산법을 이용하여 methicillin 감수성을 측정하였다. Broth microdilution 법으로 vancomycin MIC를 측정하였다. 결과 : 총 27명에서 73회의 방문과 292회의 배양이 시행되었으며 총 67주의 S. aureus가 분리되었다. 이중 64주(95.5%)가 methicillin 내성이었다. 전비공 MRSA 보균율은 VRE를 보균하고 있었던 환자는 19명 중 11명(58%)에서, VRE를 보균하지 않았던 환자는 8명 중 3명(37.5%)으로 VRE 보균자에서 보균율이 더 높았다. 총 64주의 MRSA의 vancomycin MIC의 분포는 0.5-2㎍/mL였으며 2㎍/mL를 초과하는 균주는 발견되지 않았다. 64주 중 vancomycin MIC가 1㎍/mL인 균주가 54주(84.4%)로 가장 많았으며 2㎍/mL 6주(9.4%), 0.5㎍/mL 4주(6.3%)의 순이었다. 결론 : VRE 환자에서 S. aureus의 보균율은 비보균자 보다 높았으며 분리된 균주들은 대부분 MRSA였다. VRSA 균주는 발견되지 않았다. Background : The transfer of vancomycin resistance from vancomycin-resistant enterococci (VRE) to Staphylococcus aureus has been predicted. The purpose of this study is to determine the prevalence and antibiotics resistance of S. aureus among patients colonized with VRE. Methods : Between January 2001 and December 2001, a prospective study was performed at Pusan National University Hospital on 27 patients. Surveillance swabs from nasal cavity, axilla, perineum, and rectum were obtained at weekly intervals. Methicillin susceptability of S. aureus was determined by oxacillin disk diffusion test and minimum inhibitory concentration (MIC) for vancomycin by microdilution broth test. Results : Total of 292 swab cultures were performed and 67 S. aureus isolates were collected. 64 isolates (95.5%) were resistant to methicillin. The prevalence of nasal MRSA carrier in 19 patients colonized with VRE was higher than that in 8 patients not colonized with VRE (58% vs. 37.5%). In 64 MRSA isolates, MIC (㎍/mL) for vancomycin ranged from 0.5 to 2. No isolates with MIC >2 ㎍/mL were observed. MIC of 1 ㎍/mL was shown (observed) in 54 isolates, 2 ㎍/mL in 6 isolates, and 0.5 ㎍/mL in 4 isolates. Conclusion : The prevalence of S. aureus with colonization of VRE is higher than that without colonization of VRE. Most of S. aureus isolates were resistant to methicillin. VRSA isolates were not observed.

Poster Session : Phorbol 12-myristate 13-acetate protects against necrotic cell death in response to death receptor-Ligation

( Hee Sun Byun ),( Kyeong Ah Park ),( Min Ho Won ),( Kuem Jin Yang ),( Sang Hee Shin ),( Long Zhen Piao ),( Jin Young Kwak ),( Ji Won Lee ),( Jong Sun Park ),( Jeong Ho Seok ),( Zheng Gang Liu ),( Gan 한국생화학분자생물학회 (구 한국생화학회) 2006 생화학분자생물학회 춘계학술발표논문집 Vol.2006 No.-

Quick and Simple Biodosimetry to Estimate the Absorbed Dose of Victims in Accidental Multi-Casualties

Sun-Hee Do,Da-Hee Jeong,Il-Hwa Hong,Si-Yoon Ryu,Kyu-Shik Jeong,Dong-Mi Kwak,Sung-Ho Kim,Chang-Mo Kang,Tae-Hwan Kim 한국실험동물학회 2006 Laboratory Animal Research Vol.22 No.4

Most biodosimetric studies of unanticipated radiation exposure fail to specify cause and effect because researchers have not been able to quantity a valid dosage for many victims in multi-casualties. To date, there has not been an effective technique responsible for all the requirements of ideal biodosimetry. In order to determine whether the apoptosis induction assay could be used to estimate an early dose prediction of the whole body or a part of it within 12 h after accidental radiation exposure, we examined apoptotic lymphocytes after ⁶⁰Co γ-rays within the dose range of 0.25 to 1 Gy. Apoptotic cell death rose steeply at low doses up to 1 Gy and radiation induced rapid changes in peripheral lymphocytes at all dose levels. These data suggest that apoptosis may play an important role in the homeostasis of radiosensitive target organs by the removal of damaged cells. The dose-response curves were linear-quadratic between the frequency of apoptotic lymphocytes and the dose. In addition, there were significant peaks for apoptosis at 4 and 6 h after irradiation, and the morphological findings of the irradiated lymphocytes were typical apoptotic cells that were rarely observed in the non-irradiated controls.

Ankle Evertor Strength of Healthy Subjects in Different Ankle and Toe Positions

Sun-hee Ahn,Hyun-a Kim,Jun-hee Kim,Kyung-tae Kwak,Oh-yun Kwon 한국전문물리치료학회 2019 한국전문물리치료학회지 Vol.26 No.3

Background: Ankle evertor muscles are important for preventing lateral ankle sprain. Since, the evertor muscles cross the ankle and toe joints, the position at which the ankle evertor muscle strength is measured is important. However, no studies have previously investigated the effect of ankle and toe positions on the strength of the ankle evertor muscle. Objects: This study is aimed to determine the effect of various ankle and toe joint positions on the strength of the ankle evertor muscles in healthy subjects. Methods: Eighteen healthy subjects participated in this study. Isometric ankle evertor strength of the dominant leg was determined in each subject in different ankle and toe positions (dorsiflexion (DF) with toe extension (TE), DF with toe flexion (TF), plantar flexion (PF) with TE, and PF with TF). A 2 by 2 repeated analysis of variance (ANOVA) was used to determine the difference in the evertor strength between the ankle positions (PF and DF) and toe positions (TE and TF). Results: The results indicate that there was no significant ankle position by toe position interaction effect (p=.83). However, the ankle evertor strength was significantly increased in the ankle DF position than in the PF position (p<.01), and the ankle evertor strength during eversion with TE was significantly higher than eversion with TF (p<.01). Conclusion: The findings of this study suggest that clinicians should consider the ankle and toe positions when measuring the muscle strength and during performance of selective muscle strengthening exercises of the ankle evertor muscles.

Phorbol 12-Myristats 13-Acetate Protects against Tumor Necrosis Factor (TNF)-Induced Necrotic Cell Death by Modulating the Recruitment of TNF Receptor 1-Associated Death Domain and Receptor-Interacting Protein into the TNF Receptor 1 Signaling Complex : Implication for the Regulatory Role of Protein Kinase C

Bun, Hee-Sun,Park, Kyeong-Ah,Won, Min-Ho,Yang, Keum-Jin,Shin, Sang-Hee,Piao, Longzhen,Kwak, Jin-Young,Lee, Zee-Won,Park, Jong-Sun,Seok, Jeong-Ho,Liu, Zheng-Gang,Hur, Gang-Min 충남대학교 암연구소 2007 암연구소 업적집 Vol.6 No.-

Protein kinase C (PKC) triggers cellular signals that regulate proliferation or death in a cell- and stimulus-specific manner. Although previous studies have demonstrated that activation of PKC with phorbol 12-myristate 13-acetate (PMA) protects cells from apoptosis induced by a number of mechanisms, including death receptor ligation, little is known about the effect or mechanism of PMA in the necrotic cell death. Here, we demonstrate that PMA-mediated activation of PKC protects against tumor necrosis factor (TNF)-induced necrosis by disrupting formation of the TNF receptor (TNFR)1 signaling complex. Pretreatment with PMA protected L929 cells from TNF-induced necrotic cell death in a PKC-dependent manner, but it did not protect against DNA-damaging agents, including doxorubicin (Adria-mycin) and camptothecin. Analysis of the upstream signaling events affected by PMA revealed that it markedly inhibited the TNF-induced recruitment of TNFR1-associated death domain protein (TRADD) and receptor-interacting protein (RIP) to TNFR1, subsequently inhibiting TNF-induced activation of nuclear factor-_(k)B and c-Jun NH_(2)-terminal kinase (JNK). However, JNK inhibitors do not significantly affect TNF-induced necrosis, suggesting that the inhibition of JNK activation by PMA is not part of the antinecrotic mechanism. In addition, PMA acted as an antagonist of TNF-induced reactive oxygen species (ROS) production, thereby suppressing activation of ROS-mediated poly(ADP-ribose)polymerase (PARP), and thus inhibiting necrotic cell death. Furthermore, during TNF-induced necrosis, PARP was significantly activated in wild-type mouse embryonic fibroblast (MEF) cells but not in RIP-/- or TNFR-associated factor 2-/- MEF cells. Taken together, these results suggest that PKC activation ensures effective shutdown of the death receptor-mediated necrotic cell death pathway by modulating formation of the death receptor signaling complex.