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Serratia marcescens에서 cAMP receptor protein(CRP)유전자의 클로닝 및 해석
유주순,문종환,정수열,김혜선,최용락 東亞大學校附設遺傳工學硏究所 1999 遺傳工學硏究 Vol.- No.6
전사조절인자로서 잘 알려져 있는 cAMP receptor protein(CRP)은 cAMP와 DNA에 결합하는 특별한 활성을 가지고 있으며, cAMP-CRP complex를 형성하여 수많은 유전자의 발현조절에 관여한다. 이러한 측면에서 cAMP-CRP의 조절은 어떤 면에서 총체적 조절체계라고까지 한다. 본 연구는 Serratia 균주에서 crp유전자의 분자적 특성 및 cAMP에 의한 발현조절을 받는 분자기구를 해석하고자 유전자를 클로닝하고 발현을 확인하였다. MacConkey배지에서 maltose를 탄소원으로 충분히 이용하지 못하는 대장균 TP2139(△crp,△lac)를 숙주로 이용하고, 염색체DNA를 library로 작성하여 얻은 형질전환체 약 일만개의 콜로니에서 red colony를 나타내는 5종류 의 양성 클론을 얻었다. 이들 클론을 Southern방법으로 확인한 결과 3kb의 단편을 가진 pCKB12클론이 crp유전자를 coding하고 있음을 확인하였다. glpD-lacZ 융합 plasmid인 pLDC6 의 BamHI부위에 pCKB12의 3kb 단편을 삽입시킨 재조합 plasmid pLDC 6-Scrp를 작성하여, 클로닝된 Serratia 의 crp 유전자가 대장균에서 유전자 전사조절에 미치는 영향을 확인한 결과 cAMP-CRP 복합체 형성에 의한 전사조절 기능이 확인되어졌다. One of the better-characterized transcription factor of E. coli is the cAMP receptor protein(CRP) and the CRP binds cAMP and DNA. The cAMP-CRP complex is involved in regulation of many genes at bacteria. The cAMP-CRP regulatory element represents, in some respects, a global regulatory network. The aim of this work was to study the structure and the mechanisms controlling the expression of CRP in Serratia marcescens. We have been get 5 different clones from Serratia which stimulated the cells to use maltose as a sole carbon source in E. coli TP2139. The crp gene clone, pCKB12, was confirmed by Southern hybridization with E. coli crp gene. The location of the crp gene was determined by constructing subclones carrying various portions of pCKB12. To investigate the potential role of CRP in E. cloi, lacZ fused plasmids were constructed and investigated the β-galactosidase activity of the fused plasmid. The Serratia marcescens cAMP receptor protein can substitute the E. coli CRP in transcriptional activation at the lacZ gene. These results suggest that Serratia marcescens cAMP receptor protein complex functions to regulate several promoters in E. coli.
Risk Based Inspection 기법을 이용한 화학공장의 안전성 향상에 관한 연구
노용해,유진환,서재민,임차순,고재욱 한국산업안전학회 2002 한국안전학회지 Vol.17 No.3
The RBI technique proposed by API is composed of three steps. The qualitative RBI method can be used for the purpose of screening the components with high risk. And, the quantitative RBI method employs complex risk evaluation model for predicting component risk in a quantitative manner. The inspection program can be optimized based on the results obtained by these RBI technique. The forementioned RBI technique has been applied to a common hydrodesulfurizer unit and the technique is critically evaluated for studying its benefits and limitations, which is the main issue of this thesis. It's conducted that the RBI method can provide a method for defining and measuring the component risk, and also provide a powerful tool for managing many of the important elements of a process plant.
A recurrent inactivating mutation in RHOA GTPase in angioimmunoblastic T cell lymphoma
Yoo, Hae Yong,Sung, Min Kyung,Lee, Seung Ho,Kim, Sangok,Lee, Haeseung,Park, Seongjin,Kim, Sang Cheol,Lee, Byungwook,Rho, Kyoohyoung,Lee, Jong-Eun,Cho, Kwang-Hwi,Kim, Wankyu,Ju, Hyunjung,Kim, Jaesang,K Nature Publishing Group, a division of Macmillan P 2014 Nature genetics Vol.46 No.4
The molecular mechanisms underlying angioimmunoblastic T cell lymphoma (AITL), a common type of mature T cell lymphoma of poor prognosis, are largely unknown. Here we report a frequent somatic mutation in RHOA (encoding p.Gly17Val) using exome and transcriptome sequencing of samples from individuals with AITL. Further examination of the RHOA mutation encoding p.Gly17Val in 239 lymphoma samples showed that the mutation was specific to T cell lymphoma and was absent from B cell lymphoma. We demonstrate that the RHOA mutation encoding p.Gly17Val, which was found in 53.3% (24 of 45) of the AITL cases examined, is oncogenic in nature using multiple molecular assays. Molecular modeling and docking simulations provided a structural basis for the loss of GTPase activity in the RHOA Gly17Val mutant. Our experimental data and modeling results suggest that the RHOA mutation encoding p.Gly17Val is a driver mutation in AITL. On the basis of these data and through integrated pathway analysis, we build a comprehensive signaling network for AITL oncogenesis.
The Role of Spleen Volume in Predicting Portal Hyper-tension in Patients with Cirrhosis
( Hae Won Yoo ),( Sang Gyune Kim ),( Young Seok Kim ),( Jeongju Yoo ),( Jae Woo Park ),( Yong Seok Lim ),( Jae Myeong Lee ),( Gab Jin Cheon ),( Jae Young Jang ),( Young Don Kim ),( Soung Won Jeong ),( 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1
Aims: It is already well known that the size of the spleen increases as liver cirrhosis aggravates. However, there is few studies for establishing the correlation between portal hypertension and spleen volume (SV). In this study, we aimed to see the relationship between the splenic volume and hepatic venous wedge pressure (HVPG) and to find what determine the portal pressure. Methods: From May 2007 to February 2018, 337 cirrhotic patients who measured SV and HVPG simultaneously were included. SV was measured based on the ellipsoid volume calculation formula by measuring length, height, and width. Other clinical and biochemical factors which may contribute to increase portal pressure were also collected. Results: Mean HVPG was 14.2 ± 5.3 and Child-pugh class A, B, and C patients were 34%, 40%, and 16%, respectively. A significant positive correlation was noted between splenic volume and HVPG levels (r=0.127, P=0.019). Below 16mmHg of HVPG, the stronger correlation was noted between them (r=0.202, P=0.003) but this relation was not observed above 16mmHg of HVPG. We divided HVPG into various stages of portal hypertension(HVPG<5, 5≤HVPG<10, 10≤HVPG<16, HVPG≥16). The average spleen volume of each group tended to increase with increasing HVPG value. Mean splenic volume for each group was 320.74cm3, 464.78 cm3, 501.06 cm3, 525.94 cm3 respectively. But there was no significant difference in the mean spleen volume between the groups., HVPG was correlated with presence of ascites (P=0.038) as well as SV (P=0.043) after adjusted by MELD, Na, albumin, platelet count. Conclusions: The spleen volume tended to increase with increasing HVPG and this association was stronger when HVPG was below 16 mmHg. More studies with larger numbers are needed to predict the portal pressure non-invasively.
( Hae Won Yoo ),( Young Seok Kim ),( Sang Gyune Kim ),( Jeongju Yoo ),( Jae Woo Park ),( Yong Seok Lim ),( Gab Jin Cheon ),( Jae Young Jang ),( Young Don Kim ),( Soung Won Jeong ),( Sae Hwan Lee ),( B 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1
Aims: The Baveno VI guidelines proposed that esophagogastroduodendoscopy(EGD) for screening esophageal varices(EV) can be avoided if liver stiffness(LS) measured by transient elastography(TE) is less than 20 kPa and platelet count is greater than 150,000 cells/μL. However while validation of TE is well proven, 2D-SWE has not been sufficiently validated for EV prediction. The aim of this study is to predict the presence of EV by non-invasive tools combined with 2D-SWE and to compare the diagnostic capabilities with TE. Methods: Between January 2015 and October 2017, 258 patients with compensated advanced chronic liver disease(cACLD) who underwent 2D-SWE and EGD consecutively were enrolled. The AUROC was calculated to evaluate the accuracy of the prediction for the presence of EV using 2D-SWE, model combining 2D-SWE and platelet count(PC), liver stiffness to spleen/platelet score (LSPS) score and platelet-spleen ratio (PSR) score. 177 patients who underwent simultaneous TE examination were on the subgroup analysis was done. Results: The mean age was 56.8±10.7 years and most common etiology was chronic hepatitis B (45.0%). 83.1% of patients were in Child class A. Prevalence of all-size varices was 41.1%. 2D-SWE alone has good ability to discriminate varices (AUROCs : 0.750, 95% CI, 0.690 to 0.809). Model combining 2D-SWE with platelet count and LSPS using 2D-SWE has better discriminative ability for varices and AUROCs were 0.793 (95% CI, 0.738 to 0.848) and 0.813 (95% CI, 0.760 to 0.865) respectively. For 177 patients who performed TE and 2D-SWE simultaneously, there was no difference in predictive abilities when other factors such as albumin, bilirubin, ALT, Platelet count, hemoglobin, spleen diameter were adjusted in multivariate analysis. Conclusions: 2D-SWE combined with platelet count and LSPS seemed to be useful to predict EV. In addition, 2D-SWE has similar diagnostic performance with TE for predicting presence of EV.
실무연구(實務硏究) : 민사(民事) 항소심(抗訴審) 판결(判決)의 작성(作成) 방식(方式) 개선(改善)을 위한 제안(提案)
유해용 ( Hae Yong Yoo ) 법조협회 2005 法曹 Vol.54 No.4
판결서는 소송의 대상과 재판과정, 그리고 법원의 판단을 최종적으로 정리한 종국적 답변이고, 그로써 분쟁의 법률적 종결을 선언하는 것이므로, 엄격한 사실인정과 치밀한 법리에 기초하여 신중하게 작성할 필요가 있다. 하지만 민사분쟁 해결 시스템 전반의 능률을 높이고 좀더 충실한 재판을 실현하기 위해서는 한정된 업무시간을 효율적으로 배분하는 것이 불가피하므로, 판결서 작성에 쏟는 시간과 노력을 줄여 진실발견을 위한 충실한 심리와 적정한 결론의 도출에 투입하는 것이 바람직하다. 그 동안 법원은 ``판결서 간이화``(판결 작성에 소요되는 시간·노력의 절감)를 통해 ``심리 충실과 소송 촉진``이라는 목적을 달성하기 위하여 여러 차례에 걸쳐 판결서 개선작업을 추진하였는데, 「쓰기 쉽고 읽기 편한 판결」을 지향하는 쪽으로 개선이 이루어져왔다. 2002년 7월부터 시행된 개정 민사소송법은 1심 판결 중 無辯論 판결과 자백간주 또는 공시송달에 의한 판결에는 이유 기재를 대폭 간소화하고, 일반 판결에서도 반드시 당사자의 주장, 그 밖의 공격·방어방법의 「전부」에 관한 판단을 표시할 필요는 없도록 하여 판결서 간이화를 도모할 수 있는 여지를 더욱 확대하였다. 한편, 법원 내부 전산망을 통한 판결서 정보 공유와 抗訴審 判決集 公刊 등을 통한 판결서의 일반 공개는 기존의 판결 작성 관행에 변화를 불러올 계기가 되고 있다. 이러한 상황 변화에 따라 민사 항소심 판결서는 1심 판결과 결론 및 판단 이유가 같은 사건에서는 인용판결을 활용하거나 요점만 설명하는 방식으로 간략하게 작성하는 대신, 先例的 가치가 있거나 사회적 관심도가 높은 사건에서는 외국처럼 상세하게 판단 근거를 밝힐 필요가 있다. 이 글에서는 판결서 개선을 위한 우리나라와 외국의 다양한 시도들을 간략하게 살펴본 다음, 민사 항소심 판결의 특수성을 감안한 개선 방안으로, 引用判決의 활용 확대, 쟁점 중심의 압축된 이유 구성, 쟁점정리절차를 통한 불필요하거나 무익한 주장의 사전 정리, 항소이유에 대한 판단 위주의 이유 구성 등을 제안하고, 실제 사례를 참고로 한 몇 가지 이유 기재 방식을 소개한다.