심한 Respiratory Syncytial Virus 감염증과 선천성 면역에 관련된 유전적 소인에 관한 연구 : Mannose Binding Lectin 유전자 다형성

최은화,김희섭,윤보영,최승은,나송이,김동호,박기원,이환종,Choi, Eun Hwa,Kim, Hee Sup,Yun, Bo Young,Choi, Seung Eun,Nah, Song Yi,Kim, Dong Ho,Park, Ki Won,Lee, Hoan Jong 대한소아감염학회 2006 Pediatric Infection and Vaccine Vol.13 No.1

목 적 : 본 연구는 어린 연령의 소아에서 선천성 면역력의 중요한 인자인 Mannose Binding Lectin(MBL2) 유전자의 다형성 및 일배체형의 분포를 심한 respiratory syncytial virus(RSV) 감염증 환자군과 정상대조군에서 분석함으로써, MBL2 유전자의 다양성이 소아 RSV 감염증의 발생과 예후에 기여하는지를 밝히고자 하였다. 방 법 : 1993년부터 2000년까지 7년 동안 서울대학교 어린이병원 소아과에서 RSV 감염증으로 치료받은 심한 RSV 감염증 환아 99명을 환자군, 건강한 성인 224명을 대조군으로 선정하였다. 유전형 분석은 선천성 면역의 중추적 역할을 하는 MBL2 유전자의 promoter -221 X/Y와 exon 1의 구조 변이의 분포를 환자대조군에서 비교하여 분석하였다. 결 과 : 환자군 99명의 중앙 연령은 9.5개월로, 6개월 미만 48명(49%), 6~24개월 사이 39명(39%), 그리고 24개월 이상이 12명(12%)이었다. 남아는 58명(59%)이었다. 환자군에서 MBL2 대립유전자 B다형성의 빈도는 유전형 AA 60(61%), 이형접합 AB 34(34%), 그리고 유전형 BB 5(5%)로 나타났다. 환자군의 promoter -221 X/Y 유전형의 분포는 유전형 YY 84(85%), 이형접합 XY 15(15%)이었으며, 유전형 XX는 발견되지 않았다. Exon 1의 구조 변이와 promoter 변이 모두 환자군과 대조군간에 차이가 없었다. MBL2 일배체형의 분포에도 두 군간에 차이가 없었다. 결 론 : 본 연구 결과, 한국인 소아에서 MBL2 유전자의 다형성과 일배체형이 심한 RSV 감염증에 기여하는 예후 인자임을 밝히지 못하였다. 향후 전향적으로 수집한 더 많은 수의 환자군과 적절한 대조군을 통한 연구가 시행되어야 할 것으로 생각한다. Purpose : We hypothesized that mannose binding lectin gene(MBL2), a key molecule of innate immunity, may contirbute to the development and the outcome of respiratory syncytial virus(RSV) disease in early childhood. This study was performed to investigate the genetic basis of polymorphisms and haplotypes of MBL2 for RSV disease severity in Korean children. Methods : Cases with severe RSV diseases are 99 children with severe RSV lower respiratory tract infections, who were admitted to the Seoul National University Children's Hospital through 1993~2000. The control subjects consisted of 224 anonymous healthy Korean blood donors. The frequency of promoter variant(-221, X/Y) and structural variant(codon 54) were compared between the case patient group and the control subject group. Results : The mean age of patients was 11.8 months; 49% were <6 months, 39% were 6-24 months and 12% were >24 months. In the cohort of cases of severe RSV diseases, the genotypic frequencies of structural variant in codon 54 were 61% for AA, 34% for AB, and 5% for BB. Those of the promoter X/Y variant were 85% for YY and 15% for XY. There were no significant differences in overall distribution of both structural and promoter variants between the cases and the control subjects. We did not observe statistical difference in the haplotypic frequencies of MBL2. Conclusion : Common variants of MBL2 gene most likely do not contribute to the risk for severe RSV diseases in Korean children. Further genetic association studies should be conducted in a larger propsectively recruited cohort of children with RSV infection.

국내에서 분리된 Respiratory Syncytial Virus A 아군의 항원성의 변이와 G-단백 mRNA의 RT-PCR 생산물의 제한효소 처리 및 염기 서열 결정을 통한 유전자 변이의 분석

최은화,박기호,이환종,Choi, Eun Hwa,Park, Ki Ho,Lee, Hoan Jong 대한소아감염학회 1999 Pediatric Infection and Vaccine Vol.6 No.2

목 적 : Respiratory syncytial virus(RSV)는 소아 하기도 감염증의 중요한 원인의 하나이다. 연구자들은 8년간 분리된 RSV A 아군의 항원상의 다양성과 G 단백 유전자의 변이를 관찰하고자 하였다. 방 법 : 서울대학교 어린이병원에서 1990년부터 1998년까지 8년 동안 분리된 RSV A 아군 179주들을 대상으로 하였다. G 단백 유전자를 역전사 중합효소 연쇄 반응으로 증폭한 후 이 증폭 산물을 AluI, HineII, MboI, PstI, RsaI 및 TagI 등의 제한효소로 처리하여 이들의 절단 양상에 따른 유전자형들의 다양성을 파악하였다. 또, RSV에 대한 단클론 항체와의 반응 여부에 따른 항원형과 유전자형의 연관성을 검정하였다. 또한, 각 유전자형의 선택된 일부 주들의 염기 서열을 결정하여 이미 보고된 표준주 및 외국의 임상 분리주들과 비교하여 계통수 분석을 시행하였다. 결 과 : 8회의 RSV 유행기 중 A 아군은 7회에서 유행하였다. A 아군 179주의 G 단백 유전자의 역전사 중합효소 연쇄 반응 증폭 산물의 제한효소에 의한 절단 양상에 따른 유전자형은 23가지였으며 이 중 12가지는 2개 이상의 바이러스주에서 나타났다. 단클론 항체와의 반응 양상에 따른 항원형은 6가지로 분류되었으며 그 중 한가지 형이 91%를 차지하였다. 항원헝과 유전형간의 상관관계는 적었다. 우리나라에서 분리된 A 아군의 G 단백의 염기 서열은 표준주틀과 91~93%의 동질성을 보였으며, 아미노산 서열은 표준주들과 85~90%의 동질성을 보였다. G 단백 염기 서열을 토대로 한 계통수에서 우리나라에서 분리된 RSV는 각기 표준주와는 다른 부위에 속하였고 A 아군은 4개의 군을 형성하면서 1988년부터 1993년까지 영국과 스페인에서 분리된 주들과 부분적으로 연관 관계를 나타냈다. 결 론 : 결론적으로 RSV A 아군의 유전자형은 항원형보다 더욱 다양한 양상을 보이며, 우리나라에서는 표준주 및 외국의 임상 분리주와는 차이가 있는 다양한 유전자형의 RSV가 유행하고 있다. 향후 RSV의 연구에는 이러한 점이 고려되어야 할 것으로 생각된다. Purpose : Respiratory syncytial virus(RSV) is the major cause of lower respiratory tract infection in infants and young children. This study was performed to analyze antigenic and genetic variation of G protein of subgroup A RSV. Methods : One hundred seventy-nine strains isolated at the Seoul National University Children's Hospital over 8 years-period from 1990 through 1998 were analysed for antigenic and genetic variability. Analysis was made by reactivity with monoclonal antibodies raised against RSV, and by restriction mapping and, for selected strains, nucleotide sequencing following amplification of full sequence of G gene by reverse transcription-polymerase chain reaction. Results : Restriction fragment analysis of the amplified G protein gene revealed 23 restriction patterns, 12 of which included more than 2 isolate, and the most frequent genetic type comprised 30% of the strains. Indirect immunofluorescent staining with monoclonal antibodies revealed 6 antigenic types with one predominant pattern accounting for 91% of the total strains. The most frequent antigenic type had 21 restriction patterns, and some viruses with same restiction pattern had different monoclonal antibody reaction pattern. Nucleotide sequence homology of subgroup A was 91~93% between reference(A2, Long) and Korean isolates, 93~99% among Korean isolates. Maximum-parsimony analysis demonstrated that Korean isolates were distinct from reference strains and subgroup A strains were clustered in 4 groups. Conclusion : The restriction analysis pattern of G protein gene identified greater diversity within subgroup A than was seen with the monoclonal analysis and a variety of antigenic and genetic types of RSV are circulating in Korea which are different from reference strains or strains isolated from other countries.

E . coli 에서 사람 프리트롬빈 2 cDNA 의 크로닝과 발현

최은화,김용주,김종명,홍효정,한문희,김지영 ( Eun Hwa Choi,Yong Joo Kim,Jong Myoung Kim,Hyo Jeong Hong,Moon Hi Han,Ji Young Kim ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.2

A 0.6 kbp of 3`-portion of human prothrombin cDNA was isolated from a liver cDNA library of λgt 10 by in situ plaque hybridization. A cDNA fragment corresponding to the N-terminal region of prethrombin 2 was obtained from another cDNA library synthesized by reverse transcription of human liver mRNA using a specific oligonucleotide as a primer. The 48 bp cDNA sequences starting from the first nucleotide to the AvaI site of human prethrombin 2 cDNA was chemically synthesized such that prethrombin 2 can be expressed directly in E. coli expression vector system. The open reading frame(ORF) for human prethrombin 2 was cloned in pUC18 by combining the synthetic fragment and the cDNA fragments. Expression plasmids were constructed in which the ORF for human prethrombin 2 was inserted into the site downstream from the thermoinducible leftward promoter (P_L) and the cII ribosome binding site of bacteriophage λ. Upon temperature induction, the plasmids expressed 34,000 dalton polypeptide, which was accumulated to an amount corresponding to 5-10% of the total bacterial proteins. The 34,000 dalton polypeptide was identified as human prethrombin 2 by SDS-gel electrophoresis and Western blot analysis.

Cloning and expression of human prethrombin 2 cDNA in Escherichia coli

최은화,김용주,김종명,홍효정,한문희,김지영,Choi, Eun-Hwa,Kim, Yong-Joo,Kim, Jong-Myoung,Hong, Hyo-Jeong,Han, Moon-Hi,Kim, Ji-Young 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.2

A 0.6 kbp of 3'-portion of human prothrombin cDNA was isolated from a liver cDNA library of ${\lambda}gt$ 10 by in situ plaque hybridization. A cDNA fragment corresponding to the N-terminal region of prethrombin 2 was obtained from another cDNA library synthesized by reverse transcription of human liver mRNA using a specific oligonucleotide as a primer. The 48 bp cDNA sequences starting from the first nucleotide to the AvaI site of human prethrombin 2 cDNA was chemically synthesized such that prethrombin 2 can be expressed directly in E. coli expression vector system. The open reading frame(ORF) for human prethrombin 2 was cloned in pUC18 by combining the synthetic fragment and the cDNA fragments. Expression plasmids were constructed in which the ORF for human prethrombin 2 was inserted into the site downstream from the thermoinducible leftward promoter $(P_L)$ and the cII ribosome binding site of bacteriophage ${\lambda}$. Upon temperature induction, the plasmids expressed 34,000 dalton polypeptide, which was accumulated to an amount corresponding to 5-10% of the total bacterial proteins. The 34,000 dalton polypeptide was identified as human prethrombin 2 by SDS-gel electrophoresis and Western blot analysis. 사람 간 cDNA library로부터 프로트롬빈 mRNA의 3'쪽에 해당하는 0.6kbp cDNA를 plaque hybridization에 의해 분리하였다. 프로트롬빈 2의 N-말단에 해당하는 cDNA 절편은 간 mRNA와 상보적인 올리고뉴클리오티드 primer를 이용하여 만든 다른 cDNA library로부터 분리하였다. 프로트롬빈 2를 E. coli에서 직접 발현시키기 위해 프리트롬빈 2cDNA의 첫 뉴클라오티드부터 Ava I 부위까지의 48bp DNA를 화학적으로 합성하였다. 합성 DNA와 cDNA 절편들을 이용하여 완전한 사람 프리트롬빈 2 번역프레임 (ORF)를 포함한 pUC18 을 클로닝하였다. 사람 트리트롬빈 2 ORF를 박테리오파지 ${\lambda}$의 $P_L$ promoter와 c II ribosome binding site 아래에 연결하여 프리트롬빈 2 발현벡터를 제조하였다. 온도에 의해 유도시킨 결과 이 플라스미드는 E. coli에서 약 34,000 dalton의 폴리펩티드를 생성하였다. 이 폴리펩티드는 전체 박테리아 단백질의 5-10% 정도로 만들어졌다. Western blot 분석에 의해 이 폴리펩티드가 사람 프리트롬빈 2 라는 것을 확인하였다.

1인주거공간을 위한 모듈러하우스의 방법론적 접근

최은화(Choi, Eun-Hwa),황희준(Whang, Hee-Joon) 대한건축학회 2016 대한건축학회 학술발표대회 논문집 Vol.36 No.2

This study started from the social issue; Single-person Household. Nowadays, architecture can"t reflect social changes immediately because of a nature of construction industry; that is huge and expensive. This study suggests new way of system housing for single person household. It divides space into small pieces and gives users a choice. Users can make a decision in size, number, and arrangement of their own space.

수두 예방접종: 세계적인 현황과 우리나라 접종 스케줄에 대한 제안

최은화 ( Eun Hwa Choi ) 대한소아감염학회 2008 Pediatric Infection and Vaccine Vol.15 No.1

Varicella is a highly infectious disease caused by the varicella zoster virus. The varicella vaccine was developed by Michiaki Takahashi in Japan in 1974. Despite the worldwide distribution of efficient vaccines, varicella vaccination policy is extremely variable from country to country. Although varicella vaccine is not currently recommended for universal vaccination in Japan, most countries throughout Europe, and developing countries, it had been introduced into Korea in 1988 and 20 years have elapsed since its use. Currently, varicella vaccine has been most extensively used in the United States where routine 2- dose vaccination program has been recently implemented for children. Recent 2-dose schedule in the United States and the availability of combination measles-rubella-varicella vaccines may lead to future varicella vaccination policy changes in many countries. With this background, this article summarizes the current status of varicella vaccination policies worldwide and presents provisional updated recommendation of varicella vaccination in Korea. (Korean J Pediatr Infect Dis 2008;15:11-18)

태반-특이유전자의 흰쥐 태반내 발현분포

최은화(Eun Hwa Choi),정채용(Chaeyong Jung),남광일(Kwang Il Nam),이승원(Seung Won Lee),배춘상(Choon Sang Bae),김백윤(Baik Yoon Kim),박성식(Sung Sik Park),안규윤(Kyu Youn Ahn) 대한체질인류학회 2011 해부·생물인류학 (Anat Biol Anthropol) Vol.24 No.1

Homeobox 유전자는 분화, 발생 및 발암 등을 유도하는 유전자들의 발현을 조절하는 많은 전사인자들을 암호화하고 이 유전자의 특징은 서로 다른 종 간에도 일정한 염기서열이 잘 보존되어 있다는 것이다. 본 연구는 homeobox 유전자계에 속하는 흰쥐의 새로운 태반-특이 유전자(rPsx)로 부터 digoxigenin이 표지된 cRNA probe를 만들어 in situ hybridization 조직화학 기법을 이용하여 임신 7.5일부터 16.5일까지의 흰쥐 태반에서 이의 발현 및 분포를 알아보고자 하였다. In situ hybridization 조직화학 소견에서 임신 10.5일에 처음으로 태반-특이유전자의 mRNA가 융모막외배엽에서 처음으로 발현되기 시작하였다. 임신 11.5일에서 hybridization signal은 미로영양막층과 거대영양막세포층에서만 관찰되었다. 임신 12.5, 13.5 및 14.5일에서는 hybridization signal은 거대영양막세포층, 해면영양막층 및 미로 영양막층에서 관찰되었다. 임신 15.5일에서는 hybridization signal이 미로영양막층과 해면체영양막층에서 관찰되었으나 거대영양막세포층에서는 관찰되지 않았다. 해면체영양막층에서의 hybridization signal은 임신 14.5일에 비해 상당히 감소하였다. 임신 16.5일에서는 hybridization signal이 미로영양막층에서만 관찰되었고 해면체영양막층이나 거대영양막세포층에서는 관찰되지 않았다. 임신 경과에 따른 hybridization signal은 임신 10.5일부터 점차 증가하여 14.5일에 최고조에 달한 후 그 이후에는 감소하는 경향이었다. 이상의 결과로 흰쥐에서 새로운 태반-특이유전자는 임신 10.5일부터 발현되기 시작하여 태반의 융모막외배엽, 미로영양막층, 해면영양막층 및 거대 영양막세포층에서만 발현되며 영양막세포의 분화 및 발육에 중요한 역할을 할 것으로 생각되었다. Homeobox genes seem to play critical roles in regulating morphogenesis, patterning, organogenesis, and differentiation. They have the conserved sequence that codes the DNA-binding domain called homeodomain. The expression and cellular localization of rPsx mRNA in rat placenta during placental development were examined by in situ hybridization histochemistry at different embryonic stages (Embryonic days 7.5~16.5). rPsx mRNA was first detected in chorionic ectoderm of placenta at E 10.5. This transcript was localized in labyrinth trophoblast and trophoblast giant cells at E 11.5. Hybridization signals were observed in labyrinth trophoblast, spongiotrophoblast, and trophoblast giant cells at E 12.5, E 13.5, and E 14.5. At E 15.5, hybridization signal was detected in labyrinth trophoblast and spongiotrophoblast but not in trophoblast giant cells. Hybridization signal was only detected in labyrinth trophoblast at E 16.5. rPsx mRNA was not detected in decidua and any tissues of the embryo from E 7.5 to E 9.5 of gestations. From these results, a new rPsx homeobox gene is first expressed at E 10.5 and detected in chorionic ectoderm, labyrinth trophblast, spongiotrophoblast and trophoblast giant cells of the placenta. This gene may play a critical role in differentiation and development of trophoblast cells.

비메탄계 휘발성유기화합물에 의한 간접 온실효과의 산출

최은화 ( Eun Hwa Choi ) 한국환경과학회 2012 한국환경과학회지 Vol.21 No.2

Indirect CO2 effect due to non-methane volatile organic compound (NMVOC) emissions from solvent and product use and fugitive NMVOC emissions from fuels in the Republic of Korea and 13 Annex I countries under United Nations Framework on Climate Change were estimated and the proportions of them to total greenhouse gas (GHG) emissions ranged from 0.092% to 0.45% in 2006. Indirect greenhouse effect (CO2, CH4, and O3) were estimated at 13 photochemical assessment monitoring sites in the Republic of Korea using concentrations of 8 NMVOCs of which indirect global warming potential (GWP) were available. The contribution of toluene to mixing ratio was highest at 11 sites and however, the contribution of toluene to indirect greenhouse effect was highest at nine sites. In contrast to toluene, the contributions of ethane, butane, and ethylene were enhanced. The indirect greenhouse effects of ethane and propane, of which ozone formation potentials are the lowest and the third lowest respectively among targeted 10 NMVOCs, ranked first and fourth highest respectively. Acetaldehyde has relatively higher maximum incremental reactivity and is classified as probable human carcinogen however, its indirect GWP ranked second lowest.

2015년 국내 중동호흡기증후군 유행 양상

최은화 ( Eun Hwa Choi ) 대한소아감염학회 2015 Pediatric Infection and Vaccine Vol.22 No.3

Since April 2012, more than 1,600 laboratory-confirmed human infections with Middle East Respiratory Syndrome Coronavirus (MERS-CoV) have been reported, occurring primarily in countries in the Arabian Peninsula; the majority in Saudi Arabia. The MERS outbreak in Korea, which began in May 2015 through the importation of a single case who had recently traveled to Bahrain, the United Arab Emirates, Saudi Arabia, and Qatar. As of November 28th, 186 secondary and tertiary cases had been reported; 38 deaths, mainly associated with underlying chronic illnesses, were reported. One case was exported to China and has been recorded as the first MERS case in China. Thirty-seven confirmed cases were associated with the index case, who was hospitalized from May 15 to May 17. Emergency room at one of the nation’s largest hospitals had been affected by hospital-to-hospital and intra-hospital transmissions of MERS-CoV, resulting in an outbreak of 90 infected patients. The vast majority of 186 confirmed cases are linked to a single transmission chain associated with health facilities. The median age of patients is 55 years, with a range of 16 to 87 years. The majority (61%) of patients are men. Twenty-five (14%) of the cases involve healthcare workers. The overall median incubation period was six days, but it was four days for secondary cases and six days for tertiary cases. There has been no evidence of airborne transmission and sustained human-to-human transmission in communities. Intensified public health measures, including contact tracing, quarantine and isolation of all contacts and suspected cases, and infection prevention and control have brought the MERSCoV under control in Korea. Since 4 July no new cases have been reported.

이혼 후 여성의 자기인식 변화 과정 연구

최은화 ( Eun Hwa Choi ),이윤화 ( Yon Hwa Lee ) 한국가족사회복지학회 2010 한국가족복지학 Vol.28 No.-

본 연구는 이혼 후 긍정적 적응으로서 여성이 경험하는 자기인식의 변화과정을 설명하는 연구이다. 연구 참여자는 이혼한지 4년에서 6년 된 여성 7명이었으며 이들을 대상으로 심층면접이 실시되었고 근거이론 접근방법의 절차를 따라 자료를 분석하였다. 분석결과 90개의 개념과 26개의 하위범주, 14개의 범주가 도출되었다. 이혼 후 여성의 자기인식 변화과정에서 중심현상은 ‘현실수용’과 ‘자립적인 삶 추구’로 나타났으며 이에 배경이 되는 인과적 조건은‘과거의 삶에 대한 재평가’와‘세상 앞의 막막함’이었다. ‘삶의 재지향’과 ‘새로운 삶에의 도전’이 중심현상을 이끌어내는 맥락적 조건이었고, ‘자기강화’와 ‘지지망 형성’의 중재적 조건을 통해 ‘일에의 몰두’,‘자기계발’,‘성과획득’등의 상호작용 전략이 나타났다. 그 결과로 이혼 전보다 변화되고 한층 성숙된 자신을 발견한 모습으로서의‘긍정적 사고로의 전환’,‘진정한 나의 재발견’,‘성장하는 나’를 인식하게 되는 것으로 자기인식 변화가 이루어지는 것으로 분석되었다. 이러한 결과를 토대로 이혼 후 여성의 긍정적 적응을 지원하기 위한 사회복지 실천방안에 대한 제언이 이루어졌다. The purpose of this study was to explore the changing process of self-awareness as positive adjustment process of divorced women. Seven women who had been divorced between 4 and 6 years participated in this study. Based on qualitative research, the research method was in-depth interview. From data analyses using the grounded theory, 90 concepts, 26 sub-categories and 17 categories were observed in open coding. `The core phenomenon` of the changing process of self-awareness of divorced women was conceptualized as `accept the reality` and `pursue an independent life`. As `the causal conditions`, `reassessing of the past life` and `embarrassing in front of the reality` were founded and `the context conditions` were conceptualized as `redirection of life` and `challenge of new life`. `Reinforcing the self` and `forming social support` were conceptualized as `the intervening conditions` and 3 categories -`totally preoccupied with work`, `self establishment` and `obtaining achievement` were conceptualized as `the interaction`. As a result of those process, `changing into positive thinking`, `finding true self` and `perceiving the growth of self` were founded as `consequences`. Based on the results, discussions and suggestions regarding social work practice and research to improve support system for the positive adjustment of divorced women were presented.