한국 노동자 계급의식의 발전에 관한 연구 : 노동자조직과의 관련성을 중심으로

김혜림 부산대학교 1990 국내석사

고용 상태가 건강 및 의료비 지출에 미치는 영향 : 검진 참여의 매개 효과를 중심으로

김혜림 서울대학교 대학원 2024 국내박사

Employment instability has increased along with changes in industrial structure and the economy, and employment status has been considered one of the social determinants of health. The health impact of employment can vary depending on the level of social protection in each country. Korea has a traditional employment-centered social security system, and the level of social protection for people without stable employment is low. This study was motivated by the question of whether the National Health Insurance, which is the only social security system that guarantees access to healthcare regardless of employment status, plays a protective role against the health effects of employment status, and in particular, what role the National Health Screening Program, which aims to improve the health of health insurance enrollees and reduce healthcare expenditures, plays in this process. The purpose of this study was to examine the impact of employment status on health and healthcare expenditures, focusing on the diseases targeted by the National Health Screening Program, and to examine the mediation and interaction effects of health screening participation on that impact. The National Health Insurance Service-National Health Information Database (2002―2022) was used as the data source, and workers in their 40s as of 2010 were included. Considering the possibility of reverse causality in the relationship between employment and health, the study population was limited to those who had no diseases under study or related diseases for the past 3 years and had maintained continuous employment. Employment status was defined as “employed,” “unemployed,” and “frequent employment changes” based on changes in health insurance eligibility since 2010. Health outcomes and healthcare expenditures were defined for cardiovascular disease in relation to general health screening and for gastric cancer in relation to cancer screening, respectively. The effect of employment status on health outcomes was analyzed using Cox’s proportional hazards model, and the effect on healthcare expenditures was analyzed using generalized linear models. To analyze the mediation and interaction effects of participation in health screening, four-way decomposition, a causal mediation analysis methodology, was applied. The main results of this study are as follows. The risk of hospitalization for cardiovascular disease and death due to gastric cancer was significantly higher among those who were unemployed or in frequent employment changes compared to those who were employed. Additionally, when a cardiovascular-related disease or gastric cancer occurred, healthcare expenditures due to the disease were significantly higher among those who were unemployed or in frequent employment changes compared to those who were employed. A mediation effect of screening participation was identified for the effects of employment status on hospitalization for cardiovascular disease, death due to gastric cancer, and healthcare expenditures for cardiovascular-related diseases. In terms of healthcare expenditures due to gastric cancer, mediation effect of screening participation was found only for the effect of frequent employment changes. The results should be interpreted with caution as they do not fully satisfy the confounding variable assumption. However, the magnitude of the mediation effect was not small enough to completely rule out the possibility of a substantial mediation effect. The interaction effect between employment status and screening participation was only found for healthcare expenditures due to cardiovascular-related disease. Those who did not participate in screening had higher healthcare expenditures for cardiovascular-related disease compared to those who did participate, and this impact was greater among those who were unemployed or in frequent employment changes compared to those who were employed. The results of this study show that health disparities between employment statuses exist even in preventable health outcomes, leading to increased healthcare expenditures. It also suggests the possibility that participation in the National Health Screening Program may play a part in this mechanism. This can be seen as an example of the inverse prevention law, where people who need benefits are less likely to receive them. The shift towards increased employment instability can impact not only individual health but also the financial burden on National Health Insurance. Therefore, further research and policy alternatives are needed to ensure that people with unstable employment can properly utilize the National Health Screening Program. 산업 구조 및 경제 변화와 함께 고용의 불안정성이 증가해왔으며, 고용 상태는 건강의 사회적 결정요인 중 하나로 간주되어 왔다. 이러한 고용 상태의 건강 영향은 각 국가의 사회적 보호 수준에 따라 다를 수 있다. 우리나라는 전통적인 고용 중심의 사회보장체계를 갖춘 국가로 고용이 안정적이지 않은 사람에게 사회적 보호의 수준도 낮은 편이다. 본 연구는 사회보장체계 중 유일하게 고용 상태와 관계없이 의료이용을 보장하고 있는 건강보험은 고용 상태에 따른 건강 영향에 있어 보호 역할을 하고 있는가에 대한 질문, 특히 건강보험 가입자의 건강 향상과 의료비 지출 절감을 목적으로 시행되고 있는 국가건강검진 사업이 이 과정에서 어떤 역할을 하고 있는가에 대한 질문에서 출발하였다. 이에 본 연구는 국가건강검진 사업 목표 질환을 중심으로 고용 상태가 건강 및 의료비 지출에 미치는 영향을 살펴보고, 그 영향에 국가건강검진 참여의 매개 및 상호작용 효과가 있는지 파악하는 것을 목적으로 하였다. 국민건강보험 국민건강정보자료(2002~2022)를 자료원으로 사용하였으며, 2010년 기준 40대 근로자를 대상으로 하였다. 고용과 건강 간의 관계에 대한 역인과관계의 가능성을 고려하여 과거 3년간 연구 대상 질환 및 관련 질환이 없었고, 고용을 연속적으로 유지한 경우로 대상자를 제한하였다. 고용 상태는 2010년 이후 건강보험 자격 변동 정보를 기반으로 ‘고용 유지’, ‘고용 상실’, ‘잦은 고용 변동’으로 분류하여 정의하였다. 건강 결과 및 의료비 지출은 일반건강검진과 관련하여 심혈관 질환, 암 검진과 관련하여 위암에 대해 각각 정의하였다. 고용 상태가 건강 결과에 미치는 영향에 대해서는 Cox 비례 위험 모형을, 의료비 지출에 미치는 영향에 대해서는 일반화 선형 모형을 적용하여 분석하였다. 국가건강검진 참여의 매개 및 상호작용 효과에 대한 분석에는 인과 매개 분석 방법론 중 하나인 four-way decomposition을 적용하였다. 본 연구의 주요 분석 결과는 다음과 같다. 고용 유지 상태에 비하여 고용 상실 또는 잦은 고용 변동 상태에 있는 경우 심혈관 질환으로 인한 입원 발생 위험과 위암으로 인한 사망 발생 위험이 유의하게 높은 것으로 나타났다. 또한, 심혈관 관련 질환 또는 위암이 발생한 경우 고용 유지 상태에 비하여 고용 상실 상태 또는 잦은 고용 변동 상태에서 해당 질환으로 인한 의료비 지출이 유의하게 높았다. 고용 상태가 심혈관 질환으로 인한 입원 발생, 위암으로 인한 사망 발생, 심혈관 관련 질환 의료비 지출에 미치는 영향에 있어 검진 참여의 매개 효과가 확인되었다. 위암으로 인한 의료비 지출에 있어서는 잦은 고용 변동 상태의 영향에 대해서만 검진 참여의 매개 효과가 일부 확인되었다. 교란 변수 가정을 완전히 만족한다고 보기 어려워 해석에 주의가 필요하나 매개 효과의 크기가 작지 않아 실질적인 매개 효과가 있을 가능성을 완전히 배제하기는 어려운 결과라 볼 수 있다. 고용 상태와 검진 참여의 상호작용 효과는 심혈관 관련 질환 의료비 지출에 대해서만 확인되었다. 일반건강검진에 미참여한 경우는 참여한 경우에 비해 심혈관 관련 질환 의료비 지출이 더 높게 발생하였는데 이러한 영향이 고용 유지 상태에 비해 고용 상실 상태와 잦은 고용 변동 상태에서 더 크게 나타났다. 본 연구의 결과는 예방 가능한 건강 결과에 있어서도 고용 상태 간 건강 격차가 나타나고 있으며, 이는 의료비 지출 증가로도 이어지고 있음을 보여준다. 또한 그 기전에 국가건강검진 참여가 일정 부분 작용하고 있을 가능성을 제시한다. 이는 혜택이 필요한 사람이 오히려 혜택을 받을 가능성이 낮은 역 예방 법칙의 사례로 볼 수 있다. 고용 불안정이 증가하는 방향으로 고용의 양상이 변화하고 있는 상황은 개인의 건강뿐만 아니라 건강보험 재정 부담에 영향을 줄 수 있다. 따라서 고용이 불안정한 사람에게도 국가건강검진 사업이 제대로 활용될 수 있도록 추가 연구와 정책 대안 마련이 필요하다.

폐경 이후 한국 여성의 건강관련 삶의 질 영향요인 : 제8기 국민건강영양조사 자료 활용

김혜림 강릉원주대학교 일반대학원 2024 국내석사

Background: The average lifespan of Korean women is 85 years, meaning that women spend approximately one-third of their life in middle age and beyond. Midlife and early menopause begin from the age of 40 years. During this phase, women experience physical, psychological, and social changes due to menopausal symptoms, induced by hormonal changes. These menopausal symptoms directly related to health-related quality of life of women after menopause make the reduce health-related quality of life. This present study was utilized data from the Korea National Health and Nutrition Survey, which comprises a representative sample, to measure the health-related quality of life of women after menopause using the Health-Related Quality of Life Instrument with 8 Item. This study also assesses the overall factors that affect the health-related quality of life of women after menopause. Purpose: The purpose of this study was to examine the differences in the health-related quality of life of women after menopause and to understand the factors that influence their health-related quality of life. Methods: This study was a secondary data analysis using original data from the 8th Korean National Health and Nutrition Survey in 2021. Composite sample statistical analysis was performed using SPSS/WIN 28.0. A composite sample frequency analysis and Rao-Scott . were conducted to understand participants’individual and environmental characteristics, biological and psychological variables, symptom and functional status, and general health perceptions. Differences in the characteristics and health-related quality of life of women after menopause were analyzed using independent t-tests presented in a general linear model of multiple sample analysis. Factors influencing health-related quality of life in women after menopause were analyzed using multiple regression analysis with a general linear model of multiple sample analyses. Results: Characteristics and health-related quality of life of women after menopause were determined by Individual characteristics such as muscle strength exercise(t=-2.541, p=.012), snoring(t=-2.026, p=.044), and symptom status. There was a significant difference in depression (t=-9.051, p<.001), anxiety (t=-6.493, p<.001), functional status for limitation of activities(t=-5.144, p<.001), and general health perceptions regarding perceived health status(t=10.84, p<.001). Factors that influenced the health-related quality of life of women after menopause were anxiety(B=-.14, p<.001), depression (B=-.08, p<.001), perceived health status (B=.068, p<.001), limitation of activities(B=-.05, p<.001), strength exercise practice(B=-.02, p=.012), and snoring(B=-.02, p=.044). and the explanatory power of these variabl es for health-related quality of life of women after menopause was 35.5%. Conclusion: Considering the factors that influence the health-related quality of life of women after menopause, we suggest educating menopausal women and developing programs that can improve their health-related quality of life. Women who are about to reach menopause should be provided with information about the physical and mental changes caused by menopause and encouraged to recognize and accept the changes caused by menopause positively. Preventive emotional support programs, steady muscle strength exercise practices, and education on the risk danger of persistent snoring in women afer menopause are needed. Women must be encouraged to be socially active after menopause, aware of the importance of positive subjective health, and develop and manage effective health plans. Menopausal women, Health-related quality of life, Snoring, KNHANES 연구배경: 한국 여성의 평균 수명은 85세로 인생의 1/3정도를 중년기 이후의 삶으로 보내게 된다. 중년기와 이른폐경이 시작되는 만 40세 이상부터 중년여성은 여성호르몬 변화로 생기는 폐경 증상으로 신체적, 심리적, 사회적 변화를 겪게 된다. 폐경으로 인한 수면장애나 신체적·정신적 증상들은 건강관련 삶의 질에 직접적인 관련이 있으며 폐경 이후 여성의 건강관련 삶의 질을 저하시킨다. 본 연구는 대표성 있는 표본을 추출할 수 있는 국민건강영양조사를 활용하여 폐경 이후 여성의 건강관련 삶의 질을 한국형 건강관련 삶의 질 측정 도구인 HINT-8을 통하여 측정하고, 건강관련 삶의 질에 영향을 미치는 요인을 확인하고자 한다. 연구목적: 본 연구의 목적은 폐경유무에 따른 여성의 건강관련 삶의 질의 차이를 확인하고, 폐경 이후 여성의 건강관련 삶의 질에 영향을 미치는 요인을 파악하기 위함이다. 연구방법: 본 연구는 국민건강영양조사 제8기 2021년도 원시자료를 이용한 이차자료 분석연구이다. 국민건강영양조사 원시자료의 표본설계에 따라 가중치를 적용하여 SPSS/WIN 28.0 프로그램을 이용하여 복합표본통계분석을 하였다. 대상자의 개인적 특성, 환경적 특성, 생물학적·생리적요인, 증상상태, 기능상태, 전반적인 건강지각을 복합표본 빈도분석하고, 폐경유무에 따른 개인적 특성, 환경적 특성, 생물학적·생리적요인, 증상상태, 기능상태, 전반적인 건강지각은 Rao-scott 로 분석하였다. 폐경 이후 여성의 특성과 건강관련 삶의 질의 차이는 복합표본분석의 일반선형모형에서 제시하는 독립 T-검정으로 분석하였다. 폐경 이후 여성의 건강관련 삶의 질 영향요인은 복합표본분석의 일반선형모형을 활용하여 다중회귀분석으로 분석하였다. 연구결과: 폐경 이후 여성의 특성과 건강관련 삶의 질은 개인적 특성에서는 근력운동 실천(t=-2.541, p=.012), 생리적요인에서는 코골이(t=-2.026, p=.044), 증상상태에서는 우울(t=-9.051, p<.001)과 불안(t=-6.493, p<.001)에서 유의한 차이가 있었다. 기능상태에서는 활동제한(t=-5.144, p<.001)이 전반적 건강지각에서는 주관적 건강상태(t=10.84, p<.001)가 유의한 차이가 있었다. 폐경 이후 여성의 건강관련 삶의 질에 영향을 미치는 요인은 연령(B=.02, p=.003), 교육수준(B=-.02, p=.021), 불안(B=-.14, p<.001), 우울(B=-.08, p<.001), 주관적 건강상태(B=.068, p<.001), 활동제한(B=-.05, p<.001), 근력운동 실천(B=-.02, p=.012), 코골이(B=-.02, p=.044) 순으로 나타났으며, 폐경 이후 여성의 건강관련 삶의 질을 35.5%를 설명하였다. 결론: 본 연구 결과인 폐경 이후 여성의 건강관련 삶의 질 영향요인을 근거로 하여 폐경 이후 여성의 교육에 대한 전반적인 방향 제시와 건강관련 삶의 질을 증진 시킬 수 있는 프로그램을 개발하고 적용할 필요가 있다. 폐경을 앞둔 여성에게는 폐경으로 인한 신체적·정신적 변화에 대한 정보를 제공하여 폐경으로 인한 변화에 대해 긍정적인 인식과 수용을 도모할 필요가 있겠으며, 폐경 이후 여성에게는 불안을 감소시키고, 우울을 조기에 예방할 수 있는 정서적지지 프로그램과 꾸준한 근력운동 실천의 필요성, 폐경 이후 여성의 지속적인 코골이 위험성에 대한 교육이 필요하겠다. 또한 폐경 이후에도 여성의 사회활동을 격려하며, 긍정적인 주관적 건강상태의 중요성을 인지하도록 하여 폐경 이후 여성의 효과적인 건강관리를 위한 계획을 수립하여 관리할 수 있도록 해야겠다.

강진만 해수 및 하수에서 분리된 대장균군의 항생제 내성 패턴 비교

김혜림 국립순천대학교대학원 2024 국내석사

The purpose of this study was to contribute to the safe production and distribution of shellfish by analyzing the levels of Coliform contamination and the antibiotic resistance of isolated strains in seawater and land-based sources in Gangjinman bay. The samples used in this study were 47 seawater samples, 10 major inland pollution sources(5 streams, 5 domestic sewage samples) and 26 sewage samples from wastewater treatment plants in Gangjinman bay. The total of 83 points of seawater and - iv -Sewage was used as the experiment subject. As a result of this experiment, Coliforms were detected in various sources: 6.4% in seawater, 80% in streams, 40% in domestic sewage, 61.5% in inflows from wastewater treatment plants, and 30.8% in outflows from wastewater treatment plants. Among major inland pollution sources, streams exhibited the highest level of Coliform contamination. we identified 24 strains of Coliform bacteria, including 3 strains of E. coli from seawater, 3 strains of E. colifrom streams, 2 strains of K. pneumoniae, and 1 strain of K. oxytoca from streams. domestic sewage yielded 2 strains of E. coli and 1 strain of K. pneumoniae. in the inflows of wastewater treatment plants, we found 5 strains of E. coli, 1 strain of C. freundii, 1 strain of K. oxytoca, and 1 strain of S. marcescens. outflows from wastewater treatment plants produced 2 strains of E. coli and 2 strains of C. freundii. Overall, 15 strains of E. coli, 3 strains of K. pneumoniae, 2 strains of K. oxytoca, 3 strains of C. freundii, and 1 strain of S. marcescens were identified in seawater, streams, domestic sewage, inflows of wastewater treatment plants and outflows of wastewater treatment plants. PCR testing for 15 isolated E. coli strains did not detected pathogenic E. coli genes in any of the strains. However, the isolated strains of K. pneumoniae, K. oxytoca, S. marcescens, and C. freundii, were all resistant to AM antibiotics and 66.7% of the isolated Coliform bacteria showed multiple antibiotic resistance. In this experiment, the isolated Coliform bacteria, C. freundii, K. pneumoniae and K. oxytoca, showed resistance to multiple antibiotics at rate of 100%, 67% and 50%, respectively. The results of antibiotic esistance experiments in E. coli indicated that the isolates were resistant to AM(100%), KF(33%), TIC(30%), C(30%), Additionally, 6.7% exhibited resistance to NA, GM, SXT antibiotics. However, all E. coli strains were found to be susceptible to AK, CZ, FEP, CTX, CTT, CIP. Overall, 53.3% of E. coli emonstrated resistance to multiple antibiotics. analyzing the antibiotic resistance patterns of isolated E. coli strains, E. coliL1-E and L3-E from streams and E. coli W9-IE from a sewage treatment plant matched 66.6% with the multiple antibiotic resistance pattern of E. coli S1-E isolated from seawater. E. coli L1-E and L3-E from streams and E. coli W2-IE from a wastewater treatment plant matched 66.6% with the multiple antibiotic resistance pattern of E. coli S3-E isolated from seawater, while E. coli W9-IE matched 100%. In this study, the isolation locations of E. coli with similar antibiotic resistance patterns were not located close to each other, this makes it challenging to specify the direct source of contamination in the Gangjinman bay. evertheless, it is believed that streams and inflows from wastewater treatment plants, as sources of land-based pollution in the Gangjinman bay, may have an impact on Gangjinman bay waters. Consequently, it is deemed necessary to ensure the microbial safety of shellfish produced in Gangjinman Bay through continuous analysis of antibiotic resistance to land-based contaminants.

C. Debussy 피아노 작품 연구논문에 관한 분석

김혜림 경성대학교 일반대학원 2014 국내석사

Impressionism, which was one of late 19th century art movement originated in France, was led by a group of Paris-based artists. Impressionism derives from the title of Claude Oscar Monet(1840~1926), 《Impression: soleil levant》that the reviewer Louis Leroy(1812~1885) scathingly criticized in his article. The biggest characteristic of Impressionism is responding sensitively to depicting lights. Thus, outdoor painting was often preferred to indoors', so its swift painting method to capture landscapes changing constantly by lights makes lines dividing objects obscure, and lets light and shade and color contrast appear clearly. This expression method influenced literature and music of the time, as well as art. In music, Impressionism started from a French movement against Deusche Romantik. It grafted impressionist technique of art onto music: not solving parallel progress and discord, taboo in composition. Furthermore, music used impressionist technique by using pedal and wide dynamics to express vague tone and show various expressions. Claude Achille Debussy showed an excellent creative talent in expressing abstract images into musical notes. Though, Debussy, himself, did not greet the term, Impressionism, because he claimed that he did not have any intent to compose music emulating Impressionism Painting and tried to seek a new style, freeing from dominant German tradition just focusing on solemn and perfunctory style, and returning to French music which is light and free. Nevertheless, no matter what intention he had, his music has a lot in common with Impressionism painting style. It organized the theses, which investigated Debussy's piano works, around domestic studies. 452 studies have been done: academic essays have been done from 1971 to 2012 and dissertations have been done from 1974 to 2013. The theses from 1970s to the early 1990s generally dealt with the personal information of Debussy and characteristics of his works. After the 1990s, more papers have studied external musical factors, such as contemporary arts and literary works. It means that the scope of research has been widen, not just restricted to work. The paper organized academic essays and dissertations, done in Korea, about piano works of Debussy, a leader of French Impressionism, in chronological order and analyzed the works, actively studied and subjects of them. I hope the research on the present condition of Debussy's piano works make specialists, supposed to study his works, come up with the new subject.

나노 현탁 하이드로겔 시스템을 이용한 덱사메타손의 고실 내 약물 전달 증진 연구

김혜림 단국대학교 대학원(천) 2024 국내석사

특발성 돌발성 난청 (ISSNHL)은 특별한 원인 없이 3일 이내에 연속된 3개의 주파수에서 30 dB 이상의 청력손실이 발생한 경우로 정의된다. ISSNHL 환자에게 명확한 근거에 기반한 치료 옵션은 없지만, ISSNHL을 포함한 내이 장애 치료에 스테로이드 화합물인 덱사메타손 (DEX)의 고실 내 투여 처방이 증가하고 있는 추세이다. 시판 덱사메타손 제제는 친수성 전구 약물 형태인 덱사메타손 인산나트륨 (DEX-SP)의 형태로, 친유성의 정원 창막 투과가 어려워 달팽이관에 유효한 치료 농도를 제공하지 못하고, 반복 투여를 해야 한다는 한계점을 가진다. 따라서 본 연구에서는 친유성 약물인 덱사메타손 (DEX)를 통해 정원 창막을 통한 약물 투과를 용이하게 하여 내이의 치료 농도를 높일 수 있는 가능성을 제공하고, 약물 체류 시간을 연장할 수 있는 나노 현탁 하이드로겔 제형을 설계하였다. 다양한 안정화제 스크리닝과 공정 및 조성 변수에 따른 영향을 확인하여 약 300 nm의 입자 크기를 가지는 나노 현탁액(NS) 제형을 설계하였고, 설계된 NS 제형과 혼화하였을 때, 물리화학적 안정성이 확보된 점증제를 스크리닝하여 hydrogel 제형을 설계하였다. 이후 설계된 제형에 대하여 입자 크기, 결정성, 점탄성 등을 평가하여 제조한 제형 모두 균일한 nm의 약물 입자 크기와 결정형을 가짐을 확인하였다. 또한, 시판중인 DEX-SP solution과 설계한 DEX formulations에 대하여 in vitro dissolution profile, in vivo drug retention 평가를 통해 DEX formulations이 DEX-SP에 비해 연장된 약물 방출 profile을 가지며, NS 제형보다 hydrogel 제형이 지속적인 약물 방출 profile을 나타냄을 확인하였다. 결과적으로 덱사메타손 나노 현탁 하이드로겔 제형이 고실 내 약물 전달을 증진할 수 있는 제형임에 대한 가능성을 확인하였다. Idiopathic sudden hearing loss (ISSNHL) is defined as a hearing loss of 30 dB or more at three consecutive frequencies within 3 days without any specific cause. Although there are no clear evidence-based treatment options for patients with ISSNHL, the prescription of intratympanic administration of the steroid compound dexamethasone (DEX) to treat inner ear disorders, including ISSNHL, is increasing. The commercially available dexamethasone preparation is in the form of dexamethasone sodium phosphate (DEX-SP), a hydrophilic prodrug form. It has difficulty penetrating the lipophilic round window membrane, so it does not provide an effective therapeutic concentration to the cochlea and has the limitation of requiring repeated administration. Therefore, in this study, we designed a nano-suspended hydrogel formulation that can increase the therapeutic concentration of the inner ear by facilitating drug penetration through the round window membrane through the lipophilic drug dexamethasone (DEX). A nano-suspension (NS) formulation with a particle size of about 300 nm was designed by screening various stabilizers and confirming the effects of process and composition variables. A hydrogel formulation was designed by screening thickeners that had physical and chemical stability when mixed with the designed NS formulation. Afterwards, the designed formulation was evaluated for particle size, crystallinity, viscoelasticity, etc., and it was confirmed that all manufactured formulations had a uniform drug particle size and crystal form. In addition, through the in vitro dissolution profile and in vivo drug remaining evaluation of the commercially available DEX-SP solution and the designed DEX formulations, DEX formulations have a prolonged drug release profile compared to DEX-SP, and the hydrogel formulation has a sustained drug release profile compared to the NS formulation. As a result, the potential of the Dexamethasone nano-suspension hydrogel formulation to enhance intratympanic drug delivery was confirmed.

도라지의 2배체와 4배체의 조직배양 및 단백질 발현양상

김혜림 충북대학교 2014 국내석사

The roots of Platycodon grandiflorum species either dried or fresh, are used as an ingredient in salads and traditional cuisine in Korea. To interpret the root proteins, a systematical and targeting analysis were carried out from diploid and tetraploid roots. The proteins were separated from the diploid and tetraploid roots using IEF in the first-dimension and 2D-PAGE in the second dimension along with LTQ-FT-ICR mass spectrometry. Using image analysis, a total of 39 differentially expressed protein spots were identified from diploid and tetraploid root under in vivo culture condition. Except two proteins, the rest of the identified proteins were confirmed as down-regulated such as Isocitrate dehydrogenase, Proteasome subunit alpha type-2-B. However, a total of 30 protein spots were identified from diploid roots and 40 protein spots from tetraploid roots under in vitro culture condition whereas the proteins identified from diploid roots were mostly confirmed as up-regulated such as NADH dehydrogenase, Glyceraldehyde-3-phosphate dehydrogenase but proteins identified from tetraploid roots showed down-regulated in nature such as Isoflavone reductase. Furthermore, a total of 58 proteins were identified from various explant (Control, root, shoot and callus) using MALDI-TOF-TOF mass spectrometry. Out of 58 protein spots, 21 protein spots were up-regulated such as ribulose-1, 5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, cysteine proteinase, E3 ubiquitin-protein ligase, xyloglucan 6-xylosyltransferase, glycolate oxidase, cytochrome P450, transcription factor PCF5 respectively. However, the identified proteins were functionally classified into various classes using PIR (http://pir.georgetown.edu) server based on molecular function. To this end, the identified proteins from diploid and tetraploid roots were mainly involved in oxireductase activity, nucleotide binding, transferase activity, metal cluster binding, protein binding, hydrolase activity, ion binding, peptidase activity, catalytic activity, lyase activity, transporter activity, structural molecular activity, ligase activity and conjugating protein ligase activity. Regarding molecular function, a total of 58 differential proteins were classified into 15 functional categories whereas most of them involved in nucleic acid binding (17%), unknown (16%), transferase activity (14%), ion binding (12%), catalytic activity (7%), transporter activity (7%), isomerase activity (7%), nucleotide binding (5%), oxidoreductase activity (3%), hydrolase activity (3%) peptidase activity (2%), monooxygenase activity (2%), ligase activity (2%), chaperon (2%) and chlorophyll binding (2%). In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in diploid and tetraploid roots of Platycodon grandiflorum.

스테로이드 성호르몬이 돼지의 지방전구세포의 증식과 분화에 미치는 영향

김혜림 충북대학교 2008 국내석사

Sex steroid hormones have been reported to affect body composition of animals by regulating the numbers and sizes of fat cells, and used as a medicine for human beings and a growth promotant for domestic animals. The current studies were conducted to determine the effect of sex steroid hormones (estrogen, testosterone, nandrolone) on proliferation and differentiation of baby pigs. The preadipocytes were isolated from the backfat of pigs with collagenase treatment and cultured in the DMEM/F-12 (Dulbecco's Modified Eagle's Medium/Nutrient Mixture F-12 HAM) media. Insulin(600ng/ml), transferrin(1ng/ml) and hydrocortisone(500ng/ml) were added to induce differentiation of pig preadipocytes. 10-9M, 10-7M and 10-5M of sex steroid hormones were treated for 4 days (day 0~day 4) for studying the effects on proliferation, for 5 days (day 2~day 7) for studying the effects on differentiation. Cell numbers were determined with wst-1 kit (Roche) and the degree of cell differentiation was determined by measuring the activity of glycerol-3-phosphate dehydrogenase of the cells. The one percent FBS in the culture medium was chosen by the study determining the best culture condition to minimize the influence of sex steroid hormones which might be included in DMEM/F-12 media. Initially, the effect of sex steroid hormones were compared in preadipocytes from 0 week and 2 weeks old female and male pigs. Sex steroid hormones had no effects on proliferation of predipocytes from new-born pigs, however they decreased proliferation of preadipocytes from 2 weeks old pigs. Nandrolone increased the differentiation of the preadipocytes from two weeks old pig. Based on these results, the extensive studies (4 male and 4 female pigs) were conducted to determine the effects of sex steroid hormones on the preadipocytes from two weeks old pigs. Sex steroid hormones showed no effect on proliferation of the preadipocytes, and testosterone and nandrolone increased differentiation of the predipocytes from both male and female pigs. The nandrolone showed the dose-response relationship. In summary, nandrolone stimulated the differentiation of preadipocytes from 0 and 2 week old pigs and showed greater effect on predipocytes from two week old pigs that have the high blood nandrolone concentration. The reason why nandrolone showed the stimulatory effect on differentiation of pig preadipocytes is not clear.

Effect of TMPO in the DNA dou-ble-strand break end resection and repair

김혜림 조선대학교 대학원 2018 국내박사

Lamina-associated polypeptide 2α (LAP2α) is a nuclear protein dynami-cally associating with chromatin during the cell cycle, but it was not clear that the mechanism involved in DNA repair pathway. In this study, we sought to elu-cidate molecular mechanism of LAP2α-related DNA double strand breaks repair. We show that depletion of endogenous LAP2α results in cellular hypersensitivi-ty and impaired DNA damage repair to IR. First, LAP2α is associated with ho-mologous recombination related protein NBS1. Comet assay and remaining γ-H2AX show that knockdown of LAP2α inhibits DNA repair. In addition, LAP2α-depleted cells show defect of homologous recombination (HR) but not non-homologous end joining (NHEJ). Moreover, LAP2α-depleted cells decrease the MRE11 and CtIP damage foci to IR. Therefore, these results suggest that LAP2α contributes to DNA end resection of the DNA damage response through in-teraction with NBS1. Second, LAP2α also interacts with the REV7. Function of REV7 as a DNA resection inhibitor and regulating DNA strand break repair, but it was not clear that the mechanism. Depletion of REV7 severely increases HR-mediated repair and 5’ end resection. REV7 is recruited to the DSBs it sup-presses the recruitment of MRE11, CtIP and EXO1 damage foci to IR. REV7-depleted cells are showed increase BRCA1 and RPA damage foci in G1 cells, suggesting REV7 play a role key regulator of DNA strand break end resection. Collectively, our findings identify that LAP2αis a critical component of the DNA end resection to DSBs through interaction of NBS1 and Rev7. LAP2α(TMPO)는 염색질과 연관되어 있는 핵단백질로 대부분의 연구가 세포주기 조절에 치우쳐 있으며 DNA 손상 복구 조절에 대한 연구는 미흡한 실정이다. 따라서 본 연구에서 우리는 DNA 이중 가닥 손상 복구에 관여하는 LAP2α(TMPO)의 분자적 메커니즘에 대해 밝히고자 한다. 먼저 yeast two hybrid 분석을 통해 LAP2α가 상동재조합 관련 단백질 NBS1, 비상동말단결합 연관단백질 Rev7과 결합함을 확인하고 LAP2α(TMPO)의DNA손상복구 연관성을 조사하였다. LAP2α(TMPO)의 기능 상실은 방사선조사에 의한 DNA 손상에 따른 세포 민감도를 증가시키고 DNA 손상 복구를 결핍 시킨다. 또한 세포 내 LAP2α(TMPO)의 결핍은 comet 분석과 γ-H2AX 염색을 통해 LAP2α(TMPO)의 감소가 DNA 손상 복구의 결함을 나타냄을 확인 하였다. LAP2α(TMPO) 의 결핍은 방사선조사에 의한 DNA 손상에 의한 MRE11 과 CtIP 손상foci 를 감소 시켰다. 이러한 연구결과를 종합해 볼 때 LAP2α(TMPO)는 NBS1 과의 결합을 통하여 상동 재조합 활성을 조절하는 단백질로서 DNA 말단 절제와 DNA 손상 반응에 관여하는 중요한 물질임을 제시한다. 다음으로 우리는 LAP2α(TMPO)와 REV7과의 관련성을 조사하였다. REV7은 DNA 절제를 억제하고 DNA 손상에 따른 복구를 조절하지만 명확한 기전이 알려져 있지 않다. REV7의 결핍은 5’절단 절제와 상동 재조합 매개 복구를 증가시킨다. REV7은 DNA 손상에 의한 MRE11, CtIP 그리고 EXO1의 foci 를 억제시킨다. 또한 REV7 이 결핍된 세포에서는 G1 단계에서 BRCA1 과 RPA foci 가 증가되는 것을 확일 할 수 있다. 우리의 연구 결과는 REV7 이 말단부 절제의 핵심 조절 인자임을 확인하였다. 따라서 본 연구 논문은 LAP2α(TMPO)가 NBS1과 REV7과의 결합을 통해 DNA손상복구와 DNA 말단부 절개를 조절하는 중요한 인자임을 시사한다.

PIG3의 비상동결합 활성조절 연구 : PIG3 is involved in the Non-Homologous End Joining through interaction with Ku70

김혜림 조선대학교 2015 국내석사

The Tumor suppressor protein p53-Inducing gene3 (PIG3) recently has been reported to be a new player in DNA damage signaling and response, but it was not clear that the mechanism involved in DNA repair pathway. In this study, we sought to elucidate molecular mechanism of PIG3-related DNA double strand breaks repair. We show that depletion of endogenous PIG3 sensitizes cells to DNA damage agents such as IR, bleomycin and UV. Comet assay and remaining γ-H2AX show that knockdown of PIG inhibits DNA repair. In addition, PIG3-depleted cells show defect of non-homologous end joining (NHEJ) but not ho-mologous recombination (HR). Moreover, PIG3 is associated with non-homologous end joining related protein including Ku70, Ku80 and DNA-PK in vivo. Especially, Ku70 directly interacts with PIG, suggesting a possible link be-tween a cancer and DNA repair, co-immunoprecipitation analyses revealed that the PIG segment comprising amino acids 201-250 is critical for PIG-Ku70 in-teraction. Therefore, these results suggest that PIG contributes to non-homologous end joining of the DNA damage response through interaction with Ku70.