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The High Production of Multimeric Angiotensin-converting-enzyme-inhibitor in E. coli

  • Park Je-Hyoen;Kim Sun-Hoi;Ahn Sun-Hee;Lee Jong-Hee;Kim Young-Sook;Lee Sang-Jun;Kong In-Soo
    • Fisheries and Aquatic Sciences
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    • v.4 no.2
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    • pp.84-87
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    • 2001
  • Multimeric angiotensin-converting-enzyme-inhibitor (ACE}) containing a trypsin cleavable linker peptide between ACEI was constructed. We made synthetic DNA coding for the ACEI peptide with asymmetric and complementary cohesive ends of linker nucleotides. A tandemly repeated DNA cassette for the expression of concatameric short peptide multimers was constructed by ligating the basic units. The resultant multimeric peptide expressed as soluble and trypsin treated peptide was shown at the same retention time with chemically synthetic ACEI by HPLC. The present results showed that the technique developed for the production of the ACEI multimers with trypsin cleavable linker peptides can be generally applicable to the production of short peptide.

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Posttranscriptional and posttranslational determinants of cyclooxygenase expression

  • Mbonye, Uri R.;Song, In-Seok
    • BMB Reports
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    • v.42 no.9
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    • pp.552-560
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    • 2009
  • Cyclooxygenases (COX-1 and COX-2) are ER-resident proteins that catalyze the committed step in prostanoid synthesis. COX-1 is constitutively expressed in many mammalian cells, whereas COX-2 is usually expressed inducibly and transiently. Abnormal expression of COX-2 has been implicated in the pathogenesis of chronic inflammation and various cancers; therefore, it is subject to tight and complex regulation. Differences in regulation of the COX enzymes at the posttranscriptional and posttranslational levels also contribute significantly to their distinct patterns of expression. Rapid degradation of COX-2 mRNA has been attributed to AU-rich elements (AREs) at its 3’UTR. Recently, microRNAs that can selectively repress COX-2 protein synthesis have been identified. The mature forms of these COX proteins are very similar in structure except that COX-2 has a unique 19-amino acid (19-aa) segment located near the C-terminus. This C-terminal 19-aa cassette plays an important role in mediation of the entry of COX-2 into the ER-associated degradation (ERAD) system, which transports ER proteins to the cytoplasm for degradation by the 26S proteasome. A second pathway for COX-2 protein degradation is initiated after the enzyme undergoes suicide inactivation following cyclooxygenase catalysis. Here, we discuss these molecular determinants of COX-2 expression in detail.

Exposure Assessment of Airborne Quartz from Earthwork Sites (토공사 현장에서 발생하는 공기 중 석영의 노출 평가)

  • Sung, Eun Chang;Bae, Hye Jeong;Jung, Jong-Hyoen;Phee, Young Gyu
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.25 no.4
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    • pp.584-590
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    • 2015
  • Objectives: The purpose of this study was to evaluate quartz concentrations in airborne total and respirable dusts. Materials: Respirable dust samples were collected using a 10 mm aluminum cyclone equipped with a 37 mm$5{\mu}m$ pore size PVC filter. Total dust samples were collected with a three stage cassette from three work sections at earthwork sites located in the South Chungcheong-do Province area. Results: The geometric means of quartz concentrations were $0.008mg/m^3$ and $0.004mg/m^3$ in total dust and respirable dust, respectively. The geometric means of quartz contents analyzed by FTIR were 3.74% in total dust and 3.16% in respirable dust. The geometric mean of quartz concentrations and contents in secondary blasting operations were higher than those in other operations. Conclusions: Given that secondary blasting operations had higher quartz concentrations, there is a need to reduce respirable dust, such as through wet operations.

재조합 효모를 이용한 항혈전 단백질 히루딘 발효 생산공정의 최적화

  • Kim, Myeong-Dong;Gang, Hyeon-A;Lee, Sang-Gi;Seo, Jin-Ho
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.99-102
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    • 2001
  • Recombinant Saccharomyces cerevisiae strains harboring various copy numbers of hirudin gene were developed to study dependency of hirudin expression level on its gene copy number. A linear relationship between the copy number of hirudin expression cassette and hirudin expression level was observed up to 10 copies. A double <5-integration vector truncated wi 디 1 the unnecessary bacterial genes before yeast transformation showed a four-fold increase in transformation efficiency and a 1.3-fold enhancement in hirudin expression level compared with a single <5 system. Gratuitous hirudin expression strain was developed by disrupting the GALl gene of S. cerevisiae. Glucose that was fed in a limited manner effectively supported cell growth and hi겨din expression by the gratuitous strain. Effects of methanol concentrations on hirudin production in recombinant Hansenula polymorpha were investigated in continuous and fed-batch cultures. At a steady-state of continuous culture, an optimum methanol concentration of 1.7 g/L was determined at a dilution rate of 0.18 $h^{-1}$ with 1.8 mg/L ${\cdot}$ h hirudin productivity.

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Design of Learning Achievement Evaluation Module of Intelligent Computer Assisted Instruction with Various Fuzzy Environment (다양한 퍼지 환경을 갖는 지능형 교수 시스템의 학습 성취도 평가 모듈 설계)

  • Won Sung-Hyun
    • Management & Information Systems Review
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    • v.2
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    • pp.311-334
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    • 1998
  • By decreasing in CPU price and development of computer assembling technology, personal computer fake a good chance to accelerate its supply. Recently, as being introduced new computing technology so called multi media, teaming assist system which is based on single media such as studying book, cassette tape, video tape, or something else is rapidly being replaced by new assist education system based on multi media in which it is operated by the personal computer. In the computer assist education system, there is an evaluation module which appraise learner's study level into the next study strategy. At the view of this point, this part is very important. In this part, there are some factors like Importance, complexity, or difficulty which commonly include fuzzy factors in our surrounding. But until now, we are still out of the level to handle the evaluation module adequately among the some studies. In this study, we would like to suggest a new module that evaluate learning achievement of ICAI which have a variety of fuzzy environment. We combine Independent fuzzy environment like importance, complexity, difficulty into making total evaluation of learner's achievement. By the result, with expressing by linguistic form, this study can provide the theoretical basis in which we will be able to carry out sentence toward evaluation among elementary school.

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An Experimental Study of Co-relation between English Pronunciation and Listening Comprehension of Korean College Students in Chungnam and Gyungnam Provinces (충남.경남지역 대학생들의 영어발음과 청해능력의 상관관계에 대한 실험적 연구)

  • Park, Hee-Suk;Kim, Jung-Sook
    • Speech Sciences
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    • v.11 no.3
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    • pp.55-68
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    • 2004
  • The purpose of this experimental study is to investigate the relationship between English pronunciation and listening comprehension of English diphthongs and low vowels of Korean college students from the Chungnam and Gyungnam provinces. Of 22 test sentences for listening comprehension, 15 sentences were recorded by native speakers and seven sentences were edited from Springboard by Oxford University Press. For the listening comprehension test, 90 subjects from two groups, Chungnam dialect speakers and Gyungnam dialect speakers, were selected. They listened to 22 sentences produced by audio cassette tape and completed a cloze exercise. By the results of this experiment, we observed that Korean collegians of Gyungnam province showed a better listening comprehension of words including front low vowels when they preceded voiced sounds than those of Chungnam province. When the back low vowel came in an open syllable, we also recognized the same result; Gyungnam province collegians showed better listening comprehension of words including back low vowels than those of Chungnam province. As the results of Hee-Suk Park & Jung-Soak Kim(2003) showed that Gyungnam province collegians pronounced the English low vowels longer than Chungnam province collegians, we discovered that there was a positive relation between English pronunciation and listening comprehension, especially in Gyungnam province collegians. However regarding words including English diphthongs we discovered almost no relation between English pronunciation and listening comprehension.

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Transmission Aspect of Methicillin-resistant Staphylococcus aureus in the Neurosurgical Intensive Care Unit by Analysing Genotype (유전자형 분석에 의한 신경외과 중환자실의 메티실린 내성 황색포도알균의 전파양상)

  • Kim, Yun-Kyung;Hong, Hae-Sook
    • Journal of Korean Academy of Nursing
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    • v.37 no.6
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    • pp.976-985
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    • 2007
  • Purpose: The aim of this study was to identify the present situation of hospital infection and route of infection by clarifying the transmission aspect of methicillin-resistant Staphylococcus aureus(MRSA) in a Neurosurgical Intensive Care Unit by analysing genotype. Methods: MRSA was cultured from twenty five patients with a tracheostomy, twenty five health care workers, and environments in the Neurosurgical Intensive Care Unit of one hospital in D city. Data was collected from December 21, 2004 to November 5, 2005. MRSA isolates representing each genotype were analyzed by spaA typing and a multiplex PCR method capable of identifying the structural type of the staphylococcal cassette chromosome mec(SCCmec) carried by the bacteria. Results: As the same genotype and gene sequence were found among health care workers, patients, and environments, it was assumed that there was cross transmission among them. Conclusion: This study suggests that first, as the hospital infection by MRSA between health care workers and patients in the Neurosurgical Intensive Care Unit was due to result of cross transmission and the relevance of transmission between them was verified, it is necessary to take preventive measures and conduct education. Secondly, development of nursing interventions and study of infection are needed. Thirdly, consistent investment in prevention against hospital infections and environmental renovation is needed.

Exposure Assessment of Airborne Dust in Manufacturing Industries Using Silicon Carbide in Korea (우리나라 실리콘카바이드 취급사업장의 공기 중 분진 노출평가)

  • Lee, Jun Jung;Phee, Young Gyu
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.21 no.3
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    • pp.177-183
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    • 2011
  • Occupational exposure to silicon carbide dust of manufacturing industries has seldom been evaluated in Korea. Accordingly, we evaluated various silicon carbide dust concentrations in the breathing zone of workers between May 2010 and July 2010. To compare silicon carbide dust concentrations, three dust samplers including the Institute of Occupational Medicine (IOM) sampler, 37mm cassette sampler, and Aluminum cyclone sampler were used. A total of 5 manufacturing industries producing abrasive and refractory materials using silicon carbide were investigated. The geometric mean concentrations were 2.04, 0.97, and $0.48mg/m^3$ in inhalable, total and respirable silicon carbide dust, respectively. The geometric mean concentrations of silicon carbide in abrasive material manufacturing industries were slightly higher than that of refractory manufacturing industries, and finishing operations were higher than that of other operations. It was found that the results of exposure assessment in airborne dust at manufacturing industries using silicon carbide in Korea showed exceeding rate to American Conference of Governmental Industrial Hygienists Threshold Limit Value ($3mg/m^3$) was 10% in respirable dust samples. Therefore, with the consideration of the close relationship between smaller dust size and the occurrence of occupational respiratory diseases, it is suggested to promulgate the new occupational exposure limit for respirable silicon carbide dust.

Gene silencing assessment for genes from recalcitrant or poorly studied plant species

  • Kamoi, Takahiro;Eady, Colin Charles;Imai, Shinsuke
    • Plant Biotechnology Reports
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    • v.2 no.3
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    • pp.199-206
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    • 2008
  • We have developed an efficient system of assessing the ability of a gene silencing cassette to silence transcripts from recalcitrant or poorly studied plant species by using a model plant as a host for the gene of interest. Tobacco plants transgenic for Lachrymatory Factor Synthase (LFS) enzyme activity from onion were first produced by introducing a CaMV 35S-onion-lfs gene construct. These plants were then subjected to a second transformation with an RNAi construct directed against the lfs gene sequence. LFS enzyme activity assay showed that the transgenic plants, containing both the lfs gene and the RNAi construct, had significantly reduced LFS activity. This observation was supported by Western analysis for the LFS protein and further validated by quantitative RT-PCR analysis that demonstrated a significant reduction in the lfs transcript level in the dual transformants. In this work, we have demonstrated that the RNAi construct is a suitable candidate for the development of a non-lachrymatory onion. Our model plant RNAi system has wide-reaching applications for assessment and targeting of plant secondary pathway genes, from poorly studied or recalcitrant plant species, that are important in the pharmacological, food and process industries.

Conditional Replication of a Recombinant Adenovirus Studied Using Neomycin as a Selective Marker

  • Xue, Feng;Qi, Yi-Peng;Joshua, Mallam Nock;Lan, Ping;Dong, Chang-Yuan
    • BMB Reports
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    • v.36 no.3
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    • pp.275-281
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    • 2003
  • An E1B-defective adenovirus, named r2/Ad carrying the neo expression cassette, was constructed by homologous recombination. The construction, selection (using neomycin as a selective marker), and propagation of the recombinant virus was performed in human embryonic kidney 293 cells (HEK 293). An in vitro study demonstrated that this recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as human glioma tumor cells (U251) and human bladder cells (EJ), but not in some cells with functional p53, such as human adenocarcinoma cells (A549) and human fibroblast cells (MRC-5). Also, based on the cytopathic effect (CPE), it was demonstrated, under identical conditions, that the U251 cells were more sensitive to r2/Ad replication than the EJ cells. In this paper, we report that r2/Ad could be very useful in studying the in vitro selective replication of E1B-defective adenovirus and has great potential in cancer gene therapy.