수동형 태양열 교실 난방장치에 관한 연구

박희용,정해관,Park, Hi-Yong,Jung, Hai-Kwan 대한설비공학회 1983 설비저널 Vol.12 No.2

The passive type solar energy heating system for the classroom was investigated. A classroom in a primary school located at Gangnam-ku, Seoul was taken as a model classroom the heat balance equation was established. The temperature in the classroom and solarium were calculated from the heat balance. at clear days, the temperature in the classroom and solarium were measured and compared with the calculated values. The calculated and measured values for the temperature agreed with, in general, in the increasing of $20\%$ range. It was found that the smaller size of solarium could Provide the increasing of energy efficiency for the classroom temperature.

마우스세포주 Balb/c 3T3 A31-1-1에서 Epigallocatechin gallate(EGCG)의 세포암화 억제효과에 대한 유전자발현 해석

정기경,서수경,김태균,박문숙,이우선,박순희,김승희,정해관,Jung, Ki-Kyung,Suh, Soo-Kyung,Kim, Tae-Gyun,Park, Moon-Suk,Lee, Woo-Sun,Park, Sue-Nie,Kim, Seung-Hee,Jung, Hai-Kwan 한국환경성돌연변이발암원학회 2006 한국환경성돌연변이·발암원학회지 Vol.26 No.4

Previous studies showed that epigallocatechin gallate(EGCG) have substantial effects of suppressing the N-methyl-N'-nitro-N-nitrosoguanidine(MNNG)-initiated cell transformation process on the bases of foci formation frequency and loss of anchorage dependency. In this study we tried to clarify the molecular mechanism of suppressing the cell transformation process. Mouse cell line balb/c 3T3 A31-1-1 was exposed 2 days to MNNG followed by 15 days 12-O-tetradecanoylphorbol-13-acetate(TPA) treatment for our transformation process. EGCG was added after the time point of 24 hours exposure to TPA and incubated for 19 days. 2029 genes were selected in our transformation process that showed fold change value of 1.5 or more in the microarray gene expression analysis covering the mouse full genome. These genes were found to be involved mainly in the cell cycle pathway, focal adhesion, adherens junction, TGE-$\beta$ signaling, apoptosis, lysine degradation, insulin signaling, ECM-receptor interaction. Among the genes, we focused on the 631 genes(FC>0.5) reciprocally affected by EGCG treatment. Our study suggest that EGCG down-regulate the gene expressions of up stream signaling factors such as nemo like kinase with MAPK activity and PI3-Kinase, Ras GTPase and down stream factors such as cyclin D1, D2, H, T2, cdk6.

산화적 스트레스에 의한 N'-methyl-N'-nitroguanidine의 유전독성증가

강진석(Jin Seok Kang),정기경(Ki Kyung Jung),서수경(Soo Kyung Suh),김주환(Joo Hwan Kim),이화옥(Hwa Ok Lee),정해관(Hai Kwan Jung),김승희(Seung Hee Kim),박순희(Sue Nie Park) 환경독성보건학회 2007 환경독성보건학회지 Vol.22 No.4

To investigate the possible enhancement of genotoxicity in stress environment, we examined the effect of genotoxic material in oxidative stress-induced condition using human cell line. Human lymphoblast cell line, TK6 was treated with hydrogen peroxide (H₂O₂) for induction of oxidative stress, and treated with N’-methyl-N’-nitroguanidine (MNNG), as a genetoxic material. We carried out MTS assay to set treatment doses. TK6 was treated with H₂O₂ as 6.75 (low dose) or 13.5 μM (high dose) for 2 h, and treated with MNNG as 0.117 (low dose), 0.234 (middle dose), 0.468 μM (high dose) for 2 h. As results, a treatment of MNNG induced DNA damage as dose dependently. And TK6 treated with H₂O₂ at low as well as high dose followed by MNNG treatment showed higher DNA damage compared to MNNG alone treated groups. Malondialdehyde, as a marker of lipid peroxidation was increased in H₂O₂ and MNNG treated groups. Real-time RT-PCR analyses for expression sion of several antioxidative enzymes showed that catalase mRNA and glutathione peroxidase 1 mRNA expression were decreased in H₂O₂ and MNNG treated groups. Taken together, we conclude that genotoxicity induced by MNNG is enhanced in a condition of oxidative stress induced by H₂O₂ and it suggests that it should be associated with induction of lipid peroxidation and decrease of antioxidant enzymes.

Di(2-ethylhexyl) phthalate에 의해 유도된 DNA 손상과 소핵 형성

김종원(Jong Won Kim),한의식(Eui Sik Han),박미선(Hai Kwan Jung),엄미옥(Mi Sun Park),김인숙(Mi Ok Eom),전혜승(In Sook Kim),정해관(Hye Seung Jun),심웅섭(Woong Seop Sim),오혜영(Hye Young Oh) 한국환경성돌연변이발암원학회 2001 한국환경성돌연변이·발암원학회지 Vol.21 No.1

Di-2-ethylhexyl phthalate (DEHP) is the most commonly used phthalate ester in polyvinyl chloride formulations including food packing and storage of human blood. DEHP is a well known as non-genotoxic carcinogen and endocrine disrupting chemical (EDC). DEHP have shown all negative results in ICH-guildeline recommended standard genotoxicity test battery. In this study, to assess the clastogenic and DNA damaging effect in human-derived tissue specific cells, DEHP was treated in human derived MCF-7 cells, HepG2 cells, LNCap cells, BeWo cells, MCF-10A cells, and female peripheral blood cells using micronucleus assay and in human<br/> breast carcinoma MCF-7 cells up to 1.28×10-²M using Comet assay. The in vitro micronucleus assay is a mutagenicity test system for the detection of chemicals which induce the formation of small membrane bound DNA fragment i.e. micronuclei in the cytoplasm of interphase cells, originated from clastogenic and/or aneugenic<br/> mechanism. The single cell gel electrophoresis assay (Comet assay) is used to detect DNA strand-breaks and alkaline labile site. In our results, DEHP increased significantly and/or dose-depentently and time-dependently micronucleus frequency at the 6 and 24 hr without metabolic activation system only in MCF-7 cells. DEHP treated with 2 hrs in MCF-7 cells using Comet assay induced DNA damage dose-depentantly.

Syrian hamster embryo 세포와 mouse embryo BalB/c 3T3 세포에서의 bisphenol A의 세포 형질전환 연구

김종원(Jong Won Kim),한의식(Eui Sik Han),박미선(Mi Sun Park),엄미옥(Mi Ok Eom),전혜승(Hye Seung Jun),민수진(Su Jin Min),김인숙(In Sook Kim),정해관(Hai Kwan Jung),심웅섭(Woong Seop Sim),오혜영(Hye Young Oh) 한국환경성돌연변이발암원학회 2001 한국환경성돌연변이·발암원학회지 Vol.21 No.1

To identify nongenotoxic carcinogen determined as negative by ICH guideline-recommended standard genotoxicity test battery; Ames test, chromosome aberrration assay, mouse lymphoma tk+/- assay, in vivo micronucleus assay, we picked bisphenol A as a model compound. In this study, we applied in vitro BalB/c 3T3 cell transformation assay and Syrian hamster embryo (SHE) cell transformation assay. Bisphenol A was treated upto 769.2 ug/ml in BalB/c 3T3 cells and upto 125 ug/ml in SHE cells. bisphenol A didn't induced morphological transformation both with one stage treatment protocol and with two stage treatement protocol. But, treated for 48 hr, Bisphenol A induced morphological transformation significantly in SHE cells.

화장품 원료의 피부자극성과 세포독성의 관련성

이은희(Eun Hee Lee),이종권(Jong Kwon Lee),김용규(Yong Kyu Kim),박기숙(Ki Sook Park),안광수(Kwang Soo Ahn),정경미(Kyoung Mi Jung),정해관(Jung Hai Kwan),이선희(Sun Hee Lee),정수연(Soo Youn Chung),홍진태(Jin Tae Hong) 대한약학회 2001 약학회지 Vol.45 No.3

To compare skin irritation and cytotoxicity of anti-wrinkle agents, we examined skin irritation of six anti-wrinkle agents (ascorbic acid, glycolic acid, all trans-retinoic acid, ginseng extract, retinol, EB) in New Zeland white rabbit. Cytotoxicity of these agents was determined by MTT [tetrasolium salt 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl tetrazolium bromide] at multi-time points in cultured HaCaT cell, a human immortalized keratinocyte cell. We then analyzed correlation between skin irritation and cytotoxicity by spearman's rank correlation analysis. All trans- retinoic acid shewed the highest primary irritation index (0.92) in skin irritation test. Being all the six agents not irritant, retinol showed the most cytotoxic agents. The correlation between skin irritation and cytotoxicity IC50 at different time point was 0.814,0.757,0.814 and 0.7 at 3,24, 48 and 72h, respectively. We also found that IC20 and IC80 of these agents showed similar correlation with skin irritation. These results therefore demonstrated that there is close correlation between skin irritation and cytotoxicity IC50 value by MTT in HaCaT cell at early time points by anti-wrinkle agents or IC20value. IC50 at earily time point or IC20 values may be reliable alter- native determinant of skin irritation.

폐암의 유전적 감수성과 흡연과의 관련성에 대한 연구

엄미옥,민수진,김종원,박미선,한의식,정해관,정원상,오혜영 식품의약품안전청 2000 식품의약품안전청 연보 Vol.4 No.-

같은 발암물질에 노출되어도 어떤 사람은 암에 걸리지만 어떤 사람은 암에 걸리지 않는 차이를 많은 연구자들은 유전적인 영향에 의한 개개인의 발암물질의 대사과정에 차이가 있기 때문인 것으로 생각하고 있다. 또한 홉연과 관련한 폐암의 발생에서 유전적 감수성을 결정하는 대표적 인자로 c밧ochrome pf50(CYPI 및 glutathione 1,-transferase(GST)와 같은 약물대사효소가 관련됨이 보고되고 있다. 따라서 년 연구에서는 한국인 폐암환자의 혈액으로부터 이들 유전자에 대한 다형성을 조사하여 폐암발생의 유전적 감수성을 밝히는 생계지표인자로서 이용될 수 있음을 연구하고자 하였다. 폐암환자와 정상인웨 혈액을 대상으로 PCR-RfLP (Retriction Fragment Length Polymorphism)방법으로 폐앙과 관련있는 5가지 다형성 대사유전자 CYPIAI, CYPfEl, G57%1, GSTTI, mEN(microsomal epoxide hyolase)를 조사하였다. 외부발암물질에 래한 체내 보호기전의 주된 효소로 알려진 C·STMI 및 GSTTI에 있어서는 null type이 각각 폐암환f.1군은 32%f12/37)와 27%(10/37), 정상대조군은 13%(2/15)와 7%(1/15)로 조사되어, GSTMI 및 GS71 null type 유전자 다형은 폐암발생과 밀접한 관련성(ORs=2.02 및 3.03) 을 갖는 것으로 생각된다. 또한 CYPfEl frf J 절단형인 herozygous mutant도 폐암과의 유의적 관련성tORs=3.84)이 나타났다. CYPIAI의 경우 정상대조군과 환자군에 있어서 wild type은 각각 48%(10/21)와 22%(8/37)이었으며, heterozygous mutant(.nfsp f 절단형)는 각각 38%(8/21)와 54%(20/37)로 분석되었고, 반응성이 높은 epoxide의 중간대사체의 형성에 관여하는 mEH의 경우에는 fftfjj J 절단위치에서의 rosine homozygous(YY) wild type과 hist,idine/tyrosine heterozygous (HY) mutant가 주로 관찰되'겄으며, 발현율은 각각 정상대조군에서 35%(7/20)와 65%(13/20), 폐암환 자군에서 32%(12/37)와 54ff(20/37)로 나타났으며, 특이하게 환자군에서는 histidine homorygous(HH) mutant가 14%fs/37) 관찰되었다. 또한 본 연구에서는 chromosomal instability를 검출할 수 있는 지표로서 혈구세포에서 느CE(Sister Chromatid Exchan깅e), CA(Chromosome Aberration) 및 CBMN(C린okinesis-blocked Micronucleus) 시험을 실시하였다. 그 결과 정상대조군과 환자군에서 세포 당 SCE 발현빈도는 각각 5.S3f2.34, 6.22±2.00으로 비슷하였으며, CA 유발율은 정상대조군 2.39%±1.52, 폐앙환자군 3.01%±2.』긴, 그리고 MN 윤발율은 정상대조군 4.83%츠3.02와 폐암환자군 13.1%± 7.93으로 환자군에서 증가륵는 것을 관찰하였다. Lung cancer is the most common neoplasm in the world and cigarette smoke is believed to be responsible for 90% of lung cancer. Many investigators have reported anassociation between genetic polymorphism of cytochromes P-450(CYPs) or gtutathione5-transferase (GSTs) and susceptibility to lung cancer. In Korea however, few studies havebeen conducted, so we have elucidated the role of genetic polymorpssms of CYPIAI, CYPfEt,fSThfl, GSTTI, and mEN in development of lung cancer. We have anatyzed genotypes ofDNA from 37 patients and ::1 controls. The GSTMI null genotype was present in 32%(12/37)of the lung cancer patients compared to 10%(2/21) of the controls. Patients with the GSTTInull genotype was present in 27%(10/37) compared to 5%(1/21) of the controls. And there is asignificant relationships between GSTMI and/or GSTTI null genotype and the development oflung cancer. The distribution of the CYPfEl frf J variant alfele in patients(49%) was presenthigher than that In control(f%), and the difference between them was significant. 22%(8/37)and 32%(12/37) of the lung cancer patients and 48%(10/21) and 35%(7/20) of the controlscarried the CYPIAI and mErE wild type, respectively These 2 genes was not significant to thedevelopment of lung cancer. Despite the emphasis on elucidating the association betweeninheritance of susceptibility genes 3nd lung cancer, biomarker studies to link the functionalactivities of these polymorpl)ic genes are an)enable to predict the risk for Bung cancer.Therefore, we have investigated the chromosomal instability of individuals as chromosomeaberration, sister chromatid exchange, and cytokinesis-blocked mlcronuclei assay. We found thatthe formation of micronuclei was significantly Increased in patients compared to controls. Theseresult suggest that cytokinesi.j-blocked micronuclei can be used as biomarker for determiningthe risk of development of lung cancer

비유전독성 발암물질의 검색에 관한 연구(Ⅱ)

김종원,한의식,박미선,엄미옥,전혜승,정해관,오혜영 식품의약품안전청 2000 식품의약품안전청 연보 Vol.4 No.-

의약품등록의 국제적합의를 위한 협의체(IC딨 . Interna금onal Conference on Harmonization of TecHnological Requirements for Registration of fharrnaceuticats for Human Use)에서 제시하는 유전폭성시험법에서 검색되지 않는 비유전독성 발암물질을 위한 새로운 시험법을 개발하기 위하여 1차년도 연구에서 이러찬 물질에 대한 검색 가능성을 나타낸 '생체외 소핵.시험'과 최근 환경변이원학계에서 새로이 그 유용성이 퍼두되고 있는 'Syrian hamster embryo (SHE) 세포형질전환 시험법'을 이용하여 알려진 두가지 좋류의 비유전독성 발암물질의 반응을 연구하였다. 생체외 소핵시험에서는 2,3,7,8-tetrachlorodibenzo-p-dioxin fTCDD)를 모델화합물로 선정하고, 사람유방 유래의 MCF-7, MCF-lOA,MDA-MB-231 세포를 사용하여 에스트로젠 수용체(ERf 존재여부와 세포의 유형에 파른 소핵형성을 연구하였으며, 등시에 에스트로젠 길항제인 타목시펜에 대한 영향을 보았다. 또한 TCDD가 MCF극 세포의 세포주기예 미치는 영향을 연구하기 위하여 소핵형성을 니-타렌 농도에서 flow cytometer를 이용한 세포주기 분석을 수행하였다. TCDD는 MCF-lOA세포 (ER negative)와 MDA-MB-231세포 (ER negative)에서는 소핵형성이 디미하거나 전혀 증가하지 않았으나, 뽀Cf-7세포 (ER positive)에서 재현성있는 소핵형성을 유발하였으며, 타목시펜 (10 nUt)은 TCDD (IpM)에 의한 소핵형성을 최고 47.3% 까지 억제하였다. 근또글 세포주기분석결과 TCDD, tamoxi(on 단독처리군은 읍성대조군과 유사하몄으나,병용처리군에서는 GO#'Gl arrest를 유의성있게 유발하였다. SHE 세포 형질전환시험법에서는 1차년도 연구에서 생체외 소핵시험에서 양성의 결과를 나타낸 bisphenoB A을 모델화합물로 하여 연구하였다. 시험조건하에서 blsphenol A(31.2SfM, 48시간 연속처리)는 세포혈질전찬을 유발하였다. 상기 연구결과로부터 비유전독성발암물질은 기존의 유전독성시험법중 생체왹 소핵시험(에스트로젠 수용체 양성인 세포주)과 세포형질전환#·1험(Syrian hamster 배자 초대배양세포)에서 시험계의 선택 여부에 따라 부분적으로. 유전독성 지표를 검색할 수 있음을 증명하였으며, 향후 본 연구결과에서 양성을 나타낸 세포의 세포주기 관련인자의 발현변확 등에 대한 작용기전 연구가 진행되어 새로운 생체지표를 이용한 시험법의 개발이 될 것이다 To identify nongenotoxic carcinogens which are not indentified by ICH (Internation Harmonization of I'echnological Requirements ·for Registration of Pharmaceuticals for Human Use) guideline-recommended standard genotoricfty test battery, two endpoints were chosen. They are in vitro micronilcleus 3ss3y which was useful to indentify for bisphenol Aand DEHP last year's study and in vitro 51'rian hamster embryo (SHE) ceBl transformationassay which was recently recognized as a useful tool to detect nongenotoxic carcinogen as wellas genotoxic carcnogen by Environmetal Mutagen Society. In the in vitro micronucleus assayusing MCF-7 cells, TCBD is all negative in standard genotoxicity test battery; Ames test,chromosome abem·ation assay, mouse 3ymphoma tk+/- assay, in vivo micronuclelts assay.therefore TCDD if picked up as a model compound- TCDD atso induced micronucleusformation. To identify the relationship between MN formation and estrogen receptor (ER),TCDO, BPA, and DEHP were studied using hfCF-7 cells (ER positive) and in MCF-tOfL cells(ER negative), MDA-MB-231'ce31s (ER negative). TCDD induced MN formation in bfCF-7celts (ER positive) but not or little induced MN fornlation in MCF-lOA cells (ER negative) andMDA-MB-231 (ER negative). BPA and DEHP didn't induced MN formation in MCF-lOA cells(ER negative). NP didn't induce MN formation in CHt cells but induced MN formation inMCF-7 cells, We also have included the effect of estrogen inhibitors, tamoxifen, against TCDDinduced MN formation. Tamoxifen inhihited TCDD-induced UtN formation up to 4f.3% in,4fCF-7 cells. in cell cycle analysis using flow cytorneter, 1'CDD (1 pM), tamcrifen (10 nM)didn't change cell cycle phase compared with that of negative control but, tamoxifen (10 nhf)with TCDD (1 pM) induced GO/Gl arrest significant]y. Inferring for the result above 4compounds (BPA, EIEHP, TCDD, NP), we concluded the in vitro micronucleus assay vsingMCF-7 cells is a good test method for detecting nongenotoxic carcinogen and identifying thegenotoxicity of endocrine distruptors. In the in vitro SHE celt transformation assay, bisphenol Ais picked up as a model compound. Treated for 48 hr, Bisphenol A induced morphotogicaltransformation significantly.'Strategy and methological approach for identification ofnonBenotoxic cartifogen in vitro'That to identify nongenotcxic carcinogens wllich are notidentified by standard genetic toxicology test battery should be included with additive endpointsIs convincing.