Gene cloning and utility phophorylation assay of a protein-fused substrate for a highly sensitive detection of cdc2 protein kinase using a radioisotope detection technique for the development of a protein biochip

Ko, Kyong-Cheol,Choi, Mi Hee,Park, Sang Hyun John Wiley Sons, Ltd. 2009 Journal of labelled compounds & radiopharmaceutica Vol.52 No.4

<P>The prototype of the cdc2 protein kinase in mammalian cells regulates its entry into mitosis by phosphorylating a group of key proteins in the major cell cycle transitions. In this study, using the mep45 gene encoding the 45 kDa major envelope protein (Mep45) of Selenomonas ruminantium, a rumen bacteria, a Mep45-fused substrate (PKTPKKAKKL-Mep45, MFS-cdc2) was cloned to detect the activity of cdc2 protein kinase. We report here on a strategy for the detection of a phosphorylation of a substrate catalyzed by cdc2 protein kinase by using a radioisotope detection technique. It is possible to constantly obtain a reasonable quantity of MFS-cdc2 for the cdc2 protein kinase assay and its cost can be as low as a synthesized peptide. Results of the study indicate that the Mep45-fused protein can be used effectively as a substrate for detecting the activity of cdc2 protein kinase and it can be used in developing a protein biochip for a high-throughput screening and also for studying protein–protein interactions. Copyright © 2009 John Wiley & Sons, Ltd.</P> <B>Graphic Abstract</B> <P>Gene cloning and utility phophorylation assay of a protein-fused substrate for a highly sensitive detection of cdc2 protein kinase using a radioisotope detection technique for the development of a protein biochip Kyong-Cheol Ko, Mi Hee Choi, Sang Hyun Park<SUP>*</SUP>Using the mep45 gene encoding the 45 kDa major envelope protein (Mep45) of Selenomonas ruminantium, a rumen bacteria, a Mep45-fused substrate (PKTPKKAKKL-Mep45, MFS-cdc2) was cloned to detect the activity of cdc2 protein kinase. We report here on a strategy for the detection of a phosphorylation of a substrate catalyzed by cdc2 protein kinase by using a radioisotope detection technique. Copyright © 2009 John Wiley & Sons, Ltd. <img src='wiley_img/03624803-2009-52-4-JLCR1579-gra001.gif' alt='wiley_img/03624803-2009-52-4-JLCR1579-gra001'> </P>

한국인 위암 조직의 E-cadherin 유전자 촉진자 과메틸화

박미경,김헌 충북대학교 의과대학 충북대학교 의학연구소 2003 忠北醫大學術誌 Vol.13 No.2

연구목적: 위암은 우리 나라에서 가장 흔한 암으로서, E-cadherin 유전자의 촉진자 과메틸화가 전체 위암 환자 위암 조직의 50% 이상에서 나타난다. E-cadherin 유전자 촉진자 과메틸화가 있는 위암에서는 E-cadherin 유전자 발현이 현저히 감소되며, 이에 따라 암의 침습과 전이가 유발될 가능성이 높다. 한편, 숙주의 감수성과 밀접한 관련이 있는 각종 유전자 다형성과 환경적 요인인 식이습관은 E-cadherin 유전자 촉진자 과메틸화를 유발할 가능성이 높다. 이에 본 연구자는 IL-1 와 GSTP1 유전자 다형성과 식이 습관이 E-cadherin 유전자 촉진자의 과메틸화에 미치는 영향을 파악하고, 궁극적으로는 위암발생에 미치는 영향을 알아보고자 하였다. 대상 및 방법 : 조직학적으로 위암으로 진단 받은 환자 110명과, 이들과 1:2로 성과 연령 을 짝지은 대조군 220명을 모집하고, 이들에 대하여 식이 습관과 음주, 그리고 흡연 등 위암의 위험요인 폭로에 대한 직접 면접조사를 실시하였다. 모든 대상자에 대하여 IL-1 와 GSTP1 유전자 다형성을 PCR-RFLP 방법으로 조사하였다. 위암 조직에서 추출한 DNA를 이용하여 methylation-specific PCR 방법으로 E-cadherin 유전자 촉진자의 과메틸화 여부를 확인하였다. 결과: 위암환자 110명 중 44명(40%)의 위암조직에서 E-cadherin 유전자 촉진자의 과메틸화가 나타났다. E-cadherin 유전자 촉진자의 과메틸화와 IL-1 와 GSTP1 유전자 다형성 사이에는 유의한 관련성이 관찰되지 않았다. 또한 IL-1 와 GSTP1 유전자 다형성 분포는 환자군과 대조군 사이에 유의한 차이를 보이지 않았다. 식이 습관과 음주 흡연 등의 변수와 위암 조직의 E-cadherin 유전자 촉진자의 과메틸화 여부와의 관련성 검정에서 돼지고기, 족발, 돈까스 등의 돼지고기류가 보호효과를 보였고 김치찌개, 매운 김치, 쑥은 위험요인으로 나타났다. 결론: 본 연구 결과 IL-1 와 GSTP1 유전자 다형성과 위암 조직의 E-cadherin 유전자 촉진자 과메틸화 사이에는 유의한 관련성을 볼 수 없었지만, E-cadherin 유전자 촉진자의 과메틸화와 식이 항목 사이에는 유의한 관련성을 관찰하였다. 이러한 결과들은 IL-1 나 GSTP1 보다는, 식이 요인 등의 생활습관이 E-cadherin 유전자 촉진자 과메틸화에 영향을 끼쳐 위암의 진행에 중요한 역할로 작용함을 시사한다. Purpose: Gastric cancer is the most common cancer in Korea. Hypermethylation of E-cadherin gene promoter is found in more than 50% of human gastric cancer tissues, and has been reported to be associated with the decreased expression of E-cadherin. The possibility of invasiveness and metastasis of gastric cancer increases in cases with the decreased expression of E-cadherin. The aim of this study is to investigate the effects of diet and genetic polymorphisms of IL-1β and GSTP1, and their interactions on the hypermethylation of E-cadherin gene promoter in gastric cancer tissues. Materials and Methods: One hundred and ten gastric cancer patients, and 220 age-and sex-matched controls were enrolled in this study. Direct interview using a semi-quantitative food frequency questionnaire (FFQ) was performed to get informations on the level of exposure to various diet. PCR-RFLP methods were used for the genotyping of IL-1β and GSTP1, and the methylation-specific PCR was applied to check hypermethylation of E-cadherin gene promoter in stomach cancer tissues. Results: Hypermethylation of E-cadherin gene promoter was detected in 40.0% of gastric cancer tissues. Pork, pettitoes, and pork cutlet were protective factors, whereas kimchi, kimchi stew, and mugwort were risk factors for the hypermethylation of E-cadherin gene promoter· in stomach cancer tissues. However, the hypermethylation of E-cadherin gene promoter was not associated with the genetic polymorphisms of IL-1β or GSTP1. Conclusion: These results suggest that the hypermethylation of E-cadherin promoter is associated with dietary factors but not with the genetic polymorphisms of IL-1β and GSTP1, and that progression of gastric cancer could be different according to the food intake pattern before the diagnosis of gastric cancer.

정신분열증 환자에서 청각 인식유발전위와 임상적 호전의 연관성에 대한 예비연구

박미경,최영,이형영 대한신경정신의학회 1999 신경정신의학 Vol.38 No.1

연구목적 : 본 연구는 정신분열증 환자에게 치료에 대한 반응을 예측할 수 있는 객관적 지표를 알아보려는 노력으로 시행되었다. 방 법 : 연구대상은 정신과 진단편람 제 4권 (Diagnostic and Statistical Manual of Mental Disorders-Ⅳ)의 정신분열증 진단기준을 만족시키는 23명의 입원환자였다. 치료 전 또는 치료 초기에 청각 인식유발전위를 시행하였으며 임상증상의 변화는 치료 전과 치료 4주 후 양성 및 음성 증후군 척도(Positive and Negative Syndrome Scale)를 통하여 측정하였다. 결 과 : 1) 양성척도의 호전정도는 P1과 N1의 잠복기와 유의한 정 상관관계를 보이고, P3 잠복기, P2의 전위와는 유의한 역 상관관계에 있었다. 2) 음성척도의 호전정도는 P3 잠복기와 유의한 역 상관 관계를 보였다. 3) 일반정신병리척도의 호전도 또한 P3 잠복기와 유의한 역 상관관계를 보였다. 결 론 : 이러한 결과는 치료전 실시한 청각 인식유발인자가 정신분열증 환자에게 임상적 호전의 예측인자로서 가치가 있음을 시사한다. 향후 많은 환자를 대상으로 지속적인 연구가 필요하다. Objectives : The purpose of this study was to see the correlation between the auditory cognitive evoked potential and the clinical improvement in schizophrenics. Method : The subjects were 23 patients with schizophrenia. Auditory cognitive evoked potential was examined at pretreatment or acute phase of treatment. The change of symptom was assessed by the PANSS(Positive and Negative Syndrome Scale) before drug treatment and in the 4th week of treatment. Results : 1) The change of positive subsclae was positively correlated with P1 latency(p〈.05), N1 latency(p〈.05) and negatively correlated with P3 latency(p〈.05), P2 amplitude(p〈.05). 2) The change of negative subscale was negatively correlated with P3 Latency(p〈.05). 3) The change of general psychopathology subscale was negatively correlated with P3 latency(p〈.05). Conclusion : These results suggest that the auditory cognitive evoked potential in schizophrenic patients could be valuable on prediction of clinical improvement. But, further studies are required to support these results.

정신분열병 환자의 인터류킨-2 생산능

김경용,김임,박주흥,은헌정,김동인,이선미 예수병원 1997 예수병원학술지 Vol.18 No.-

Electrochemical synthesis of nanosized hydroxyapatite/graphene composite powder

Vesna Mišković-Stanković,Sanja Eraković,Ana Janković,Maja Vukašinović-Sekulić,Miodrag Mitrić,Young Chan Jung,Soo Jin Park,Kyong Yop Rhee 한국탄소학회 2015 Carbon Letters Vol.16 No.4

Electrochemical synthesis was employed to prepare a novel hydroxyapatite/graphene (HAP/ Gr) composite powder suitable for medical applications as a hard tissue implant (scaffold). The synthesis was performed in a homogeneous dispersion containing Na2H2EDTA·2H2O, NaH2PO4 and CaCl2 with a Ca/EDTA/PO4 3− concentration ratio of 0.25/0.25/0.15M, along with 0.01 wt% added graphene nanosheets, at a current density of 137 mA cm−2 and pH value of 9.0. The field emission scanning electron microscopy and transmission electron microscopy observations of the composite HAP/Gr powder indicated that nanosized hydroxyapatite particles were uniformly placed in the graphene overlay. Raman spectroscopy, Fourier transform infrared spectroscopy and X-ray diffraction confirmed graphene incorporation in the HAP/Gr powder. The electrochemically prepared HAP/Gr composite powder exhibited slight antibacterial effect against the growth of the bacterial strain Staphylococcus aureus.

가덕도주변 해역 해수의 중금속 함량

노미경,차미선,박근태,손홍주,문두호,이상준 부산대학교 환경문제연구소 2000 環境硏究報 Vol.18 No.-

The quantitative analysis of heavy metals(As, Cd, Cr, Hg, Pb, Cu) in the sea water around Kaduck island was conducted during the period of March, 1998 to September, 1998. The sea water in the four regions was sampled four times and the heavy metals were analyzed. The Heavy metal concentration of the sea water around Kaduck island was As 127.7㎍/ℓ , Cd 49.28㎍/ℓ , Cr 0.462㎍/ℓ, Hg 0.111㎍/ℓ, Pb 1.713㎍/ℓ, Cu 2.716㎍/ℓ in the average value, respectively.

일부농촌지역의 농약사용실태 및 농약중독 요인 조사

이경민,송주희,장재혁,심수정,강양화,안재경,이숭호,박미영,정문호 대한보건협회 2002 대한보건연구 Vol.28 No.4

Object The usage of agricultural chemical is on the increasing. Use of pesticides has increased agricultural production. However, negative externalities from such use have increased too. Pesticide poisoning is a major public problem. The purpose if this study is to find out the relationship between keeping the safety rules or protective equipments or attending safety education or dose of pesticides use , and poisoning experience. Methods For Gu, H.M. and Pack, S.G. study , we obtained questionnaire data concerning pesticides poisoming. A person interviews on 203 were conducted in two villages in Sinmeari and woulbonri Chunchun , Kangwon province, July 2002. Result 1. 144 people(70.9%) have sprayed pesticides and the rata of experiences of pesticide poisoning was 26% among 144. Spraying time of pesticides was 1-2 hours in 63.9% of farmers. Rate of using the protective equipment was 25.7% (protective clothes), 49.3% (Mask), 48.6 (Gloves), 7.0 (Protective glasses), 64.6% (boots) respectively. 2. Chi-square and multiple logistic regression analysis showed that Duration of spraying per day (<2hr) and using of mask(protective equipment)are significantly association with poisoning experience.(x^2=5.2684 p=0.0217, CI=0.140-0.5853 OR=0.346 p=0.0211 respectively). But no association between poisoning experience and keeping safety rule, attending of safety education. 3. Spraying pesticides. Duration of spraying pesticides, reading manuals and no spraying at fatigue (keeping safety rule) are significantly association with symptom experience(x^2=14.0621 p=0.0002, x^2=7.0639 p=0.0079, CI01.395-10.950 OR=3.908 p=0.0095, CI=0.101-0.726 OR=0.271 0.0094 respectively). But no association between symptom experience and protective equipments, attending of safety education.

치위생사의 건강보험 요양급여기준과 치과진료수가기준에 대한 인지도

전미용,박경숙 중앙대학교 의과대학 간호학과 간호과학연구소 2000 중앙간호논문집 Vol.4 No.2

Korean health insurance finally established the whole national health insurance by its rapid development since the health insurance was conducted 12 years ago, and owing to the fact the meidcal usage of the people was suddenly increased. Consequently, in this study I tried to understand the degree of the knowledge after researching the degree of the recognition of the health insurance of dental sanitarians who worked in the dental meidcal organization, and to suggest the basic data about the importance of the future health insurance education and the effective educational program, I abstracted some people arbitrarily of 300 sanitarians who worked in dental clinics and dental medical centers located in Seoul from October 9th to October 28th 2000, and the I surveyed them with direct interviews, and finally I performed the statistical anaysis treatment on 228 people of those sanitarians above. The collected data were analyzed using SAS(Statistical Analysis System) program, and the conclusion I could find through the study is as follows. 1. The degree of the knowledge about the recuperation payment of the dental sanitarian was 3.07 as an average score, and in descending order they can be displayed as the outside-hospital prescription, recuperation payment process, and the objection request for the treatment fee, and on the other hand, the lowest one was the computation of the recuperation payment period. 2. The degree of the knowldege about the standard of the dental treatment cost fee of the dental sanitarian was 3.44 as an average score, and in descending order they can be displayed as the rapid daily charging treatment, muscular tube medical care, and the mouth surgery in the field of the dental disease treatment, and in ascending order the lower articles were the organ regeneration inducement treatment, bone organs deformity correcting treatment, and the dental muscular excision treatment in the field of the surgeon. 3. The degree of the knowldege in each general characteristic of the dental sanitarians was higher in those who were old enough and had a lot of clinical experiences and there was a lot of cases that the sanitarians themselves requested in the electronic documentary interchange way about the standard of the recuperation payment and the standard of the dental treatment cost fee. When summarizing those results above, in order to increase the degree of the knowledge about the health insurance of the sanitarians, the consistent and systematic payment education should be performed. Especially, because in the group which was consisted of people who were insufficiently aged and had unsatisfactory clinical experiences, the degree of knowledge about the health insurance was shown to be low, the work guid book about the recruit dental sanitarian should be prepared, and the systematic payment education and information service are thought to be necessary. Additionally, when educating about the payment to the dental sanitarian, the payment eudcation about the concrete contents such as the dental treatment and the calculation guideline for the remedy fee, medication prescription, and medicine compounding fee should be performed. The health insurance cost fee is bing changed continuously, and the payment education of the dental sanitarians should be performed continuously, and additionally because there is no insurance-inspecting dental sanitarian who charges the inspection work professionally, the cultivation of the specially trained employees and the institutional backup for increasing the participation degree in the payment education should be executed.

정신분열병 환자의 인터류킨-2 생산능

이선미,박주홍,김동인,은헌정,김임,김경용 大韓神經精神醫學會 1996 신경정신의학 Vol.35 No.4

급성기 및 관해상태의 정신분열병 환자와 정상 대조군의 IL-2 생산능을 비교함으로써, 정신분열병의 여러 발병원인 중 자가면역 병인론의 증거인 세포성 면역방응의 저하 및 질병상태와 관련한 세포성 면역기능의 변화를 일부 한국인 정신분열병 환자에서 조사에서 조사하고자 하였다. 방 법 : DSM-IV의 정신분열병 진단기준에 적합한 급성기 환자 19명(남 : 10, 여 : 9)과 관해상태의 환자 21명(남 :12, 여: 9) 및 정상대조군 40명(남 : 20, 여 : 20)을 연구대상으로 하였다. 피험자로부터 채혈한 혈액에서 임파구를 분리하여 phytohemagglutinin으로 T임파구를 자극하고 세포배양하여 증식시킨후 IL-2immunoassay kit을 이용하여 T임파구의 IL-2 생산능을 측정하였다. 결 과 : IL-2 생산능은 성별에 관계없이 정신분열병 환자군에서 정상 대조군에 비하여 유의하게 감소하였다. (t=5. 97, p<0.001). 질병상태에 따라 급성기 정신분열병 환자군 및 관해상태 정신분열병 환자군으로 구분하여 정상대조군의 IL-2 생산능과 비교한 결과, IL-2 생산능이 정상 대조군에서 가장 높았고 다음은 관해상태 환자군이었으며 급성기 환자군에서 가장 낮았다. 세 군 사이에는 모두 유의한 차이가 있었다. (F=35.35, p<0.001). 결 론: 일부 한국인 정신분열병 환자에서 질병상태와 유의하게 관련된 세포성 면역기능의 저하가 관찰되며, 이는 정신분열병의 다양한 원인중 자가면역 기전에 의한 발병을 시사한다. Objects : This study was designed to examine the decreased cellular immune response and the change in cellular immunity related to to clinical status in Korean schizophrenic patients. Methods : The subjects were 19 acute schizophrenics and 21 remitted schizophrenics who met the DSM-IV criteria of schizophrenia and 40 healthy volunteers as the normal control. After the lymphocyte was separated from blood which had been drawn from the subjects and then T lymphocyte was stimulated by PHA and proliferated by cell culture, the examiner measured IL-2 productivity by IL-2 immunoassay kit. Results : IL-2 productivity of the schizophrenics was significantly lower than that of the normal controls. There was no significant difference between male and female in two groups. In the comparison by clinical status, acute schizophrenic group was the lowest, remitted schizophrenic group was next, and normal control group was the highest in the IL-2 productivity, with statistically significant difference among the groups(p<0.001). Conclusion : We suggest that there is a decrease in the cellular immune response which is significantly related to clinical status in some Korean schizophrenics. This finding supports that onset of schizophrenia may be the result of autoimmune mechanism, which is one of the various etiologic factors in schizophrenia.

VRE 감염증의 임상상, 미생물학적 특성 분석 및 유전적 관련성 비교 : 전국 6개 병원에서 분리된 VRE를 중심으로

정희진,박정원,엄중식,하경임,신종희,김정미,이혜수,박영희,김우주,박승철 대한감염학회 2002 감염 Vol.34 No.5