7 research outputs found
RNA editing modulates the transcriptional repression activity of CtBP1
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ผ๋ฌธ (์์ฌ)-- ์์ธ๋ํ๊ต ๋ํ์ : ์๊ณผํ๊ณผ, 2012. 8. ์คํ๋.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 โ
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TABLE OF CONTENTS โ
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LIST OF FIGURES โ
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LIST OF TABLES โ
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ABBREVIATIONS โ
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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
๊ตญ๋ฌธ์ด๋ก 40Maste
Studies on some mechanisms of the enzyme secretion of exocrine pancreas in dogs
์ํ๊ณผ/๋ฐ์ฌ[ํ๊ธ]
[์๋ฌธ]
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
์คํ์๊ฐํธ ์ ๊ณต/์์ฌ[ํ๊ธ]๋ณธ ์ฐ๊ตฌ๋ ๊ด์๋๋งฅ ์ฐํ์ ํ 4๊ฐ์์์ 8๊ฐ์์ด ์ง๋ ํ์์ ์์ ์ ์ค์ฆ๋, ์์ ํ ์ง๊ฐ๋ ๊ฑด๊ฐ์ํ ๋ฐ ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง์ ์ ๋์ ์ด๋ค๊ฐ์ ์๊ด๊ด๊ณ๋ฅผ ์์๋ณด๊ธฐ ์ํ ์์ ์ , ํก๋จ์ ์๊ด๊ด๊ณ ์ฐ๊ตฌ๋ก์ ๊ด์๋๋งฅ ์ฐํ์ ํ์์ ํ๋ณต๊ธฐ ๊ฐํธ๊ด๋ฆฌ์ ๊ธฐ์ด์๋ฃ๋ฅผ ์ ๊ณตํ๊ธฐ ์ํ์ฌ 2006๋
4์ 1์ผ๋ถํฐ 2006๋
6์ 5์ผ ๊น์ง ๊ฒฝ๊ธฐ๋ ์์ฌ์ ์ผ ๋ํ๋ณ์์์ ๊ด์๋๋งฅ ์ฐํ์ ์ ๋ฐ๊ณ ์์ ํ 4๊ฐ์์์ 8๊ฐ์์ ๊ธฐ๊ฐ์ด ๊ฒฝ๊ณผํ 61๋ช
์ ๋์์ผ๋ก ์ํํ์๋ค. ์์ง๋ ์๋ฃ๋ SPSS for window 12.0 ํ๋ก๊ทธ๋จ์ ์ด์ฉํ์ฌ ๊ธฐ์ ํต๊ณ(์ค์, ๋ฐฑ๋ถ์จ, ํ๊ท ๊ณผ ํ์คํธ์ฐจ, ์ค๊ฐ๊ฐ), ANOVA, Pearson correlation coefficient๋ก ๋ถ์ํ์๋ค.๋ณธ ์ฐ๊ตฌ๋ฅผ ํตํ์ฌ ์ป์ ๊ฒฐ๊ณผ๋ ๋ค์๊ณผ ๊ฐ๋ค.1. ์ผ๋ฐ์ ํน์ฑ์ ๋ฐ๋ฅธ ์์ ์ ์ค์ฆ๋๋ ์์ธก์ฌ๋ง ์ํ๊ณผ ์์ธก๋์กธ์ค ์ํ์ด ์ง์
์ ๋ฌด, ๊ฒฐํผ์ํ, ๋ฐฐ์ฐ์ ์ ๋ฌด, ๋๊ฑฐํํ, ์ฃผ ๊ฐํธ์ ๊ณต์, ์น๋ฃ๋น ๋ถ๋ด์์ ๋ฐ๋ผ ์ ์ํ ์ฐจ์ด๋ฅผ ๋ณด์๋ค(p<0.05).2. ์ผ๋ฐ์ ํน์ฑ์ ๋ฐ๋ฅธ ์์ ํ ์ง๊ฐ๋ ๊ฑด๊ฐ์ํ ๋ฐ ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง์ ์ง์
์ด ์๊ณ ํ๋ณต๊ธฐ ๊ต์ก์ด ๋์์ด ๋ ๊ฒฝ์ฐ ์์ ํ ์ง๊ฐ๋ ๊ฑด๊ฐ์ํ ๋ฐ ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง์ด ๋์๋ค(p<0.05).3. ์ง๋ณ ๋ฐ ์น๋ฃ๊ด๋ จ ํน์ฑ์ ๋ฐ๋ฅธ ์์ ์ ์ค์ฆ๋๋ ์์ ์ ์ํ์์ธ ์ค ํก์ฐ, ๊ธ์ฑ ์ฌ๊ทผ๊ฒฝ์์ ์ง๋จ ๋ฐ์ ๊ฒฝ์ฐ์ ์์ ์ ๋๋๋งฅ ๋ด ํ์ ํํ(IABP)๋ฅผ ๊ฑฐ์นํ ๊ฒฝ์ฐ์์ ์ ์ํ๊ฒ ๋๊ฒ ๋ํ๋ฌ๊ณ ์์ ํ ์ฃผ์ํฉ๋ณ์ฆ๊ณผ ๊ธฐํ ํฉ๋ณ์ฆ์ด ๋ฐ์ํ ๊ฒฝ์ฐ ์์ ์ ์ค์ฆ๋๋ ๋๊ฒ ๋ํ๋ฌ์ผ๋ฉฐ NYHA functional class์ ์ค์ฆ๋๊ฐ ๋์์๋ก ๋ถ๋ฏธ ์ค์ฆ๋ ํ๊ฐ๋๊ตฌ(NNERS)์ ์ค์ฆ๋๊ฐ ๋์๋ค. ์์ ์ ์ค์ฆ๋๊ฐ ๋์์๋ก ์์ ํ ์ธ๊ณต๊ธฐ๋ ์ ์ง๊ธฐ๊ฐ, ์คํ์์ค ์ฒด๋ฅ๊ธฐ๊ฐ, ์ฌ์๊ธฐ๊ฐ์ด ๊ธธ์๋ค(p<0.05).4. ์ง๋ณ ๋ฐ ์น๋ฃ๊ด๋ จ ํน์ฑ์ ๋ฐ๋ฅธ ์ง๊ฐ๋ ๊ฑด๊ฐ์ํ๋ ์์ ์ ์ํ์์ธ ์ค ํก์ฐ, ์ข์ฌ์ค ๊ธฐ๋ฅ๋ถ์ (์ข์ฌ์ค ๊ตฌํ๋ฅ ์ด ๋ฎ์ ๊ฒฝ์ฐ), ์์ ์ ๋๋๋งฅ ๋ด ํ์ ํํ(IABP)๋ฅผ ๊ฑฐ์นํ๋ ๊ฒฝ์ฐ์์ ๋ฎ๊ฒ ๋ํ๋ฌ๋ค. ์์ ํ ์ฃผ์ ํฉ๋ณ์ฆ๊ณผ ๊ธฐํ ํฉ๋ณ์ฆ์ด ๋ฐ์ํ ๊ฒฝ์ฐ๊ฐ ๊ทธ๋ ์ง ์์ ๊ฒฝ์ฐ์ ๋นํด ์์ ํ ์ง๊ฐ๋ ๊ฑด๊ฐ์ํ๊ฐ ๋ฎ์๋ค. ์์ ์ NYHA functional class์ ์ค์ฆ๋๊ฐ ์ฌํ ์๋ก ์์ ํ ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง์ด ๋ฎ์ผ๋ฉฐ ์์ ํ ์ธ๊ณต๊ธฐ๋ ์ ์ง๊ธฐ๊ฐ, ์คํ์์ค ์ฒด๋ฅ๊ธฐ๊ฐ, ์ฌ์๊ธฐ๊ฐ์ด ๊ธธ์๋ก ์์ ํ ์ง๊ฐ๋ ๊ฑด๊ฐ์ํ๋ ๋ฎ์๋ค(p<0.05).5. ์ง๋ณ ๋ฐ ์น๋ฃ๊ด๋ จ ํน์ฑ์ ๋ฐ๋ฅธ ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง์ ์์ ์ ์ํ์์ธ ์ค ์ข์ฌ์ค ๊ธฐ๋ฅ๋ถ์ ์ด ์๋ ๊ฒฝ์ฐ(์ข์ฌ์ค ๊ตฌํ๋ฅ ์ด ๋ฎ์ ๊ฒฝ์ฐ)์์ ๋ฎ๊ฒ ๋ํ๋ฌ๋ค. NYHA functional class์ ์ค์ฆ๋๊ฐ ๋์์๋ก ์์ ํ ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง์ ๋ฎ์๋ค. ์์ ํ ์ฃผ์ ํฉ๋ณ์ฆ๊ณผ ๊ธฐํ ํฉ๋ณ์ฆ์ด ๋ฐ์ํ ๊ฒฝ์ฐ๊ฐ ๊ทธ๋ ์ง ์์ ๊ฒฝ์ฐ์ ๋นํด ์์ ํ ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง์ด ๋ฎ์ ๊ฒฐ๊ณผ๋ฅผ ๋ณด์์ผ๋ฉฐ ์์ ํ ์ธ๊ณต๊ธฐ๋ ์ ์ง๊ธฐ๊ฐ, ์คํ์์ค ์ฒด๋ฅ๊ธฐ๊ฐ, ์ฌ์๊ธฐ๊ฐ์ด ๊ธธ์๋ก ์์ ํ ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง์ ๋ฎ์๋ค(p<0.05).6. ์์ ์ ์ค์ฆ๋๊ฐ ๋์์๋ก ์์ ํ ์ง๊ฐ๋ ๊ฑด๊ฐ์ํ์ ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง์ ๋ฎ์๋ค(p<0.05).7. ์์ ํ ์ง๊ฐ๋ ๊ฑด๊ฐ์ํ๊ฐ ๋์์๋ก ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง์ ๋์๋ค(p=0.000).8. ์์ ํ ํ๋ณต๊ธฐ์ ๊ฒฝํํ๋ ์ ์ฒด์ ๊ฑด๊ฐ๋ฌธ์ ๋ ๊ฐ์ด์ ๊ฐ์ ์ ๋ถํธ๊ฐ, ๋ค๋ฆฌ ๋ฐ ํ์ ์ ๊ฐ์ ์ ๋ถํธ๊ฐ, ๋ฑ, ๋ชฉ, ์ด๊นจ์ ๋ถํธ๊ฐ, ์์๋ถ์ง, ์๋ฉด์ฅ์ ์ด์ง๋ฌ์, ์จ์ฐฌ ์ฆ์์ด ์์๊ณ ์ฌ๋ฆฌ์ ๊ฑด๊ฐ๋ฌธ์ ๋ก๋ ์ฐ์ธ, ๋ถ์, ์์กด๊ฐ ์ ํ, ์คํธ๋ ์ค ๋ฑ์ ํธ์ํ์๋ค.์ด์์ ๊ฒฐ๊ณผ, ๊ด์๋๋งฅ ์ฐํ์ ํ์์ ์์ ์ ์ค์ฆ๋๋ ์์ ํ ์ง๊ฐ๋ ๊ฑด๊ฐ์ํ ๋ฐ ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง๊ณผ ์๊ด๊ด๊ณ๋ฅผ ๋ณด์ด๋ฏ๋ก ์์ ์ ์ค์ฆ๋ ๋ถ๋ฅ๋ฅผ ์ํํ์ฌ ์ค์ฆ๋์ ํน์ฑ ๋ฐ ์ ๋์ ๋ฐ๋ผ ์์ ํ ํฉ๋ณ์ฆ ์๋ฐฉ ๋ฐ ํ๋ณต๊ธฐ๊ฐ ๋จ์ถ์ ์ํ ๊ฐํธ์ค์ฌ๊ฐ ํ์ํ๋ค. ๋ํ ์์ ์ ํ๋ณต๊ธฐ ๊ต์ก์ด ์์ ํ ์ง๊ฐ๋ ๊ฑด๊ฐ์ํ ๋ฐ ๊ฑด๊ฐ๊ด๋ จ ์ถ์ ์ง์ ์ํฅ์ ์ฃผ๋ฏ๋ก ๋์์์ ๊ต์ก ์๊ตฌ๋์ ๋ฐ๋ฅธ ๊ต์ก ํ๋ก๊ทธ๋จ์ ๊ฐ๋ฐ๊ณผ ์ฐจ๋ณํ๋ ์์ง์ ๊ต์ก์ด ํ์ํ๋ค. ์ด๋ฌํ ์ธก๋ฉด์์ ์ฌํ๊ด๊ณ ์ ๋ฌธ๊ฐํธ์ฌ์ ์ญํ ๋ฐ ๊ธฐ๋ฅ์ ๋ฐํ ํ ์ ์๊ธฐ๋ฅผ ๊ธฐ๋ํ๋ค.
[์๋ฌธ]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 patientsโ 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