RNA editing modulates the transcriptional repression activity of CtBP1

학위논문 (석사)-- 서울대학교 대학원 : 의과학과, 2012. 8. 윤홍덕.RNA modification includes alternative splicing and RNA editing, which increases the diversity of transcripts and proteins. The C-terminal binding protein (CtBP), a transcriptional corepressor, has alternative splicing forms but there is no information about involving RNA editing in CtBP function. Here we found a unique transcript variant of CtBP1 with an A-to-G conversion at codon 528 in breast cancer cell lines. CtBP1 mRNA editing produces protein in which threonine-176 is changed into alanine. This CtBP1 (T176A) mutant lacks in vitro dehydrogenase activity and dimerization ability compared with wild-type CtBP1. Microarray analysis showed that many oncogenes were upregulated in CtBP-knockout mouse embryonic fibroblast. These candidate target genes were repressed by restoring wild-type CtBP1, but not by CtBP1 (T176A) mutant, indicating that CtBP1 (T176A) mutant is defective in transcriptional repression activity. We propose that RNA editing modulates the role of CtBP1 in transcription repression of oncogenes involved in breast cancer cells.ABSTRACT ⅰ TABLE OF CONTENTS ⅱ LIST OF FIGURES ⅲ LIST OF TABLES ⅳ ABBREVIATIONS ⅴ INTRODUCTION 1 MATERIALS AND METHODS 3 Cell culture and transfection 3 Plasmids 3 RNA editing analysis 3 Immunofluorescent staining 4 Protein purifications 4 In vitro GST-pull down assay 5 Dehydrogenase activity assay 5 Microarray analysis 5 RNA purification and quantitative real-time PCR 6 Lentivirus-based shRNA 6 Chromatin immunoprecipitation 7 Primers 7 RESULTS 10 1. CtBP1 mRNA editing in breast cancer cell lines 10 2. The characteristics of an edited CtBP1 protein 14 3. The change of gene expression patterns by CtBP in mouse embryonic fibroblast 19 4. Transcriptional repression activity of edited CtBP1 25 DISCUSSION 31 REFERENCES 34 국문초록 40Maste

Studies on some mechanisms of the enzyme secretion of exocrine pancreas in dogs

의학과/박사[한글] [영문] Pavlov and his associates (1888, 1893) developed experimental technics for the study of pancreatic secretion and demonstrated that stimulation of the peripheral ends of vagus nerve caused a flow of Pancreatic juice from pancreatic fistula in the unanesthetized dog, but failed to obtain similar results in acute experiments. Pavlov interpreted that vagi convey two kind of fibers to the pancreas, excitatory and inhibitory. Further he concluded that both vasoconstrictor and secretory fibers to the pancreas were present in the sympathetics. Popielski (1901), and lately Thomas (1950) has proposed a concept that local reflexes meditated through the intrinsic ganglion of the pancreas and the enteric plexus, or through the sympathetic ganglia, play in the response of the pancreas to excitants, particularly, those which increase enzyme output. On the basis of animal and human experiment Hong et al (1956, 1961) postulated the Possibility that a role of the local nerves suggested by Thomas (1950) is to release pancreas stimulating hormones from the duodenum. In order to access the role of local reflex in the regulation of enzyme secretion of the pancreas, author has studied the effects of a potent muscarinic ganglionic stimulant and its blockade or other related agents on pancreatic secretion in the presence of a secretin background in wish chloralose anesthetized dogs. Dogs between 8 and 16 kg were anesthetized with chloralose, approximately 60 mg/kg in propylene glycol after prime thiopental anesthesia (25 mg/kg). The abdominal cavity was opened, the main pancreatic duct cannulated end the accessory duct ligated. A tribe inserted into stomach for drain of gastric juice or the pylorus was ligated. All agents were injected intravenously through the polyethylene tube put in femoral vein. Seoretin (Jorpes and Mutt preparation) was infused throughout the experiment. Other agents are as follows; epinephrine and norepinephrine, new ganglionic stimulants: McN-A-343 and AHR-602, and SKF-525-A which is known to Potentiates the action of the various drugs, and ganglionic blocking agents: atropine and hexamethonium. Pancreatic juice in the presence of a secretin background was collected since there is very little flow of pancreatic juice in the fasted anesthetized dog. The pancreatic amylase, lipase and trypsin were measured by the methods previously described(Hong, 1967). Summary In dogs with chloralose anesthesia following a prime dose of thiopental the secretory response of pancreatic enzyme to secretin was studied by means of some agents active at autonomic ganglionic sites and obtained the following results. 1. Both epinephrine and norepinephrine reduced the volume response to secretin by the raising dose, however, the enzyme output was moderately increased only by the latter. 2. SKF-525-A increased the enzyme content in secretin juice. 3. Both AHR-602 and McH-A-343, muscarinic ganglionic stimulants increased the enzyme content of secretin juice and particularly the latter was a potent stimulant of enzyme secretion. 4. Both atropine and hexamethonium significantly reduced the pancreatic response to SKF-525-A, AHR-602 and McN-A-343, however, the former is more susceptible of enzyme response and the latter was of volume response. By these results it is concluded that pancreatic secretion can be elicited in part through autonomic ganglionic sites which are sensitive to both atropine and hexamethonium. However, the response of enzyme is more depend on atropine sensitive receptors and the volume response is on hexamethonium-sensitive sites. Consequently it is felt that the local reflex mechanists may explain in some extent the enzyme secretion of the pancreas, whereas, the ganglionic sites responsible for enzyme secretion appears to be cholinergic nature of atropine specific.restrictio

(The) relationship between post operative perceived health status, health-related quality of life and pre oper

중환자간호 전공/석사[한글]본 연구는 관상동맥 우회술 후 4개월에서 8개월이 지난 환자의 수술 전 중증도, 수술 후 지각된 건강상태 및 건강관련 삶의 질의 정도와 이들간의 상관관계를 알아보기 위한 서술적, 횡단적 상관관계 연구로서 관상동맥 우회술 환자의 회복기 간호관리에 기초자료를 제공하기 위하여 2006년 4월 1일부터 2006년 6월 5일 까지 경기도 소재의 일 대학병원에서 관상동맥 우회술을 받고 수술 후 4개월에서 8개월의 기간이 경과한 61명을 대상으로 시행하였다. 수집된 자료는 SPSS for window 12.0 프로그램을 이용하여 기술통계(실수, 백분율, 평균과 표준편차, 중간값), ANOVA, Pearson correlation coefficient로 분석하였다.본 연구를 통하여 얻은 결과는 다음과 같다.1. 일반적 특성에 따른 수술 전 중증도는 예측사망 위험과 예측뇌졸중 위험이 직업유무, 결혼상태, 배우자 유무, 동거형태, 주 간호제공자, 치료비 부담자에 따라 유의한 차이를 보였다(p<0.05).2. 일반적 특성에 따른 수술 후 지각된 건강상태 및 건강관련 삶의 질은 직업이 있고 회복기 교육이 도움이 된 경우 수술 후 지각된 건강상태 및 건강관련 삶의 질이 높았다(p<0.05).3. 질병 및 치료관련 특성에 따른 수술 전 중증도는 수술 전 위험요인 중 흡연, 급성 심근경색을 진단 받은 경우와 수술 전 대동맥 내 풍선펌프(IABP)를 거치한 경우에서 유의하게 높게 나타났고 수술 후 주요합병증과 기타 합병증이 발생한 경우 수술 전 중증도는 높게 나타났으며 NYHA functional class의 중증도가 높을수록 북미 중증도 평가도구(NNERS)의 중증도가 높았다. 수술 전 중증도가 높을수록 수술 후 인공기도 유지기간, 중환자실 체류기간, 재원기간이 길었다(p<0.05).4. 질병 및 치료관련 특성에 따른 지각된 건강상태는 수술 전 위험요인 중 흡연, 좌심실 기능부전(좌심실 구혈률이 낮은 경우), 수술 전 대동맥 내 풍선펌프(IABP)를 거치했던 경우에서 낮게 나타났다. 수술 후 주요 합병증과 기타 합병증이 발생한 경우가 그렇지 않은 경우에 비해 수술 후 지각된 건강상태가 낮았다. 수술 전 NYHA functional class의 중증도가 심할수록 수술 후 건강관련 삶의 질이 낮으며 수술 후 인공기도 유지기간, 중환자실 체류기간, 재원기간이 길수록 수술 후 지각된 건강상태는 낮았다(p<0.05).5. 질병 및 치료관련 특성에 따른 건강관련 삶의 질은 수술 전 위험요인 중 좌심실 기능부전이 있던 경우(좌심실 구혈률이 낮은 경우)에서 낮게 나타났다. NYHA functional class의 중증도가 높을수록 수술 후 건강관련 삶의 질은 낮았다. 수술 후 주요 합병증과 기타 합병증이 발생한 경우가 그렇지 않은 경우에 비해 수술 후 건강관련 삶의 질이 낮은 결과를 보였으며 수술 후 인공기도 유지기간, 중환자실 체류기간, 재원기간이 길수록 수술 후 건강관련 삶의 질은 낮았다(p<0.05).6. 수술 전 중증도가 높을수록 수술 후 지각된 건강상태와 건강관련 삶의 질은 낮았다(p<0.05).7. 수술 후 지각된 건강상태가 높을수록 건강관련 삶의 질은 높았다(p=0.000).8. 수술 후 회복기에 경험하는 신체적 건강문제는 가슴절개선의 불편감, 다리 및 팔의 절개선의 불편감, 등, 목, 어깨의 불편감, 식욕부진, 수면장애 어지러움, 숨찬 증상이 있었고 심리적 건강문제로는 우울, 불안, 자존감 저하, 스트레스 등을 호소하였다.이상의 결과, 관상동맥 우회술 환자의 수술 전 중증도는 수술 후 지각된 건강상태 및 건강관련 삶의 질과 상관관계를 보이므로 수술 전 중증도 분류를 시행하여 중증도의 특성 및 정도에 따라 수술 후 합병증 예방 및 회복기간 단축을 위한 간호중재가 필요하다. 또한 수술 전 회복기 교육이 수술 후 지각된 건강상태 및 건강관련 삶의 질에 영향을 주므로 대상자의 교육 요구도에 따른 교육 프로그램의 개발과 차별화된 양질의 교육이 필요하다. 이러한 측면에서 심혈관계 전문간호사의 역할 및 기능을 발휘 할 수 있기를 기대한다. [영문]The purpose of the study was to exam the relationship between post operative perceived health status & health related quality of life and pre operative disease severity in patients undergone Coronary Artery Bypass Grafting.This is descriptive and cross-section correlative study to see the relationship between post operative perceived health status & health related quality of life and pre operative disease severity in CABG patients.This study has been done from April 1, 2006 to June 5, 2006 in one university hospital in Geyonggi-Do. Total 61 patients having had the CABG with 4 months to 8 months of duration after surgery have been participated for this study.Collected data has been analyzed using SPSS for window 12.0 program with descriptive statistics (the real number, percentage, average, standard deviation and mid-point), ANOVA and Pearson correlation coefficient.The results of the study are as follows;1. The higher the degree of serious illness before surgery is, the lower the perceptive health status after surgery is (p<0.05).2. The higher the degree of serious illness before surgery is, the lower the health related quality of life after surgery is (p<0.05).3. The higher the condition of the perceptive health status after surgery is, the higher the health related quality of life is (p=0.000).4. Physical health problems during recovery phase after surgery were discomfort of chest incision line, discomfort of leg & arm incision lines, discomfort of back, neck, shoulder, lack of appetite, sleeping problem, dizziness, being short of breath. Psychological problems were depression, anxiousness, self-esteem decline and stress.In conclusion, there were significant relationship between of post operative perceived health status & health related quality of life and pre operative disease severity in patients undergone CABG. These results suggest that nursing intervention is needed according to the characteristics and degrees of the seriousness for complication prevention and shortening convalescence. In addition, as the education about convalescence before surgery affects on perceptive health status and health related quality of life, it is needed to develop an education program reflecting patients’ demand and to offer differentiated and good quality education. Thus, the cardiovascular nurse specialist and/or nurse practitioner should develop the educational program in the post operative management of CABG patients.ope