• Title/Summary/Keyword: 고속스크리닝

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Screening for inhibitory effect on nine CYP isoforms by 20 herbal medications (고속 스크리닝 기법을 이용한 한약제제의 cytochrome P45O 저해능 탐색)

  • Kim, Hyun-Mi;Liu, Kwang-Hyeon
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.334-339
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    • 2007
  • We evaluated the potential of 20 herbal medications (HMs), commonly used in Korea, to inhibit the catalytic activities of several cytochrome P450 (CYP) isoforms. The abilities of 500 ${\mu}g/ml$ of aqueous extracts of 20 HMs to inhibit phenacetin O-deethylation (CYP1A2), coumarin 6-hydroxylation (CYP2A6), bupropion hydroxylation (CYP2B6), rosiglitazone hydroxylation (CYP2C8), tolbutamide 4-methylhydroxylation (CYP2C9), S-mephenytoin 4'-hydroxylation (CYP2C19), dextromethorphan O-demethylation (CYP2D6), chlorzoxazone 6-hydroxylation (CYP2E1), and midazolam 1'-hydroxylation (CYP3A) were tested using human liver microsomes. The HMs Woohwangcheongsimwon suspension and Hwanglyeonhaedok-Tang strongly inhibited CYP2B6 and CYP2D6 isoform activity, respectively. These results suggest that some of the HMs used in Korea have potential to inhibit CYP isoforms in vitro. Although the plasma concentrations of the active constituents of the HMs were not determined, some herbs could cause clinically significant interactions because the usual doses of those individual herbs are several grams of freeze-dried extracts.

Manufacturing Protein-DNA Chip for Depigmenting Agent Screening (전사인자 저해제 통한 미백제 탐색용 단백질 칩 제작)

  • Han Jung-Sun;Kwak Eun-Young;Lee Hyang-Bok;Shin Jlung-Hyun;Baek Seung-Hak;Chung Bong-Hyun;Kim Eun-Ki
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.479-483
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    • 2004
  • An attempt was made to develop a proteinchip for screening of MITF (microphthalmia transcription factor) inhibitor. Binding of MITF to E-box causes transcription of several pigmenting genes including tyrosinase gene. We investigated binding of MITF and its DNA binding site (E-box) using a protein-DNA chip with various detection methods including flurorescence (Cyt3). SPR (surface plasmon resonance) and SPRi (surface plasmon resonance imaging). A fusion protein (MITF-Maltose Binding Protein) was attached on the glass plate by chemical modification. An inhibitory synthetic DNA oligomer, artificially designed based on the E-box sequence, inhibited the binding of MITF and E-box. These results showed the potentials of flurorescence-based MITF protein chip as a microarray for high throughput screening (HTS) system of depigmenting agents.

A Fast Screening Algorithm for On-Line Transient Stability Assessment (온라인 과도안정도 판정을 위한 상정사고 고속 스크리닝 알고리즘 개발)

  • Lee, Jong-Seock;Yang, Jung-Dae;Lee, Byong-Jun;Kwon, Sae-Hyuk;Nam, Hae-Kon;Choo, Jin-Boo;Lee, Koung-Guk;Yun, Sang-Hyun;Park, Byung-Cheol
    • The Transactions of the Korean Institute of Electrical Engineers A
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    • v.50 no.5
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    • pp.225-233
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    • 2001
  • SIME(SIngle Machine Equivalent) method has been recognized as a useful tool to determine transient stability of power systems. In this paper, SIME method is used to develop the KEPCO transient stability assessment (TSA) tool. A new screening algorithm that can be implemented in SIME method is proposed. The salient feature of the proposed screening algorithm is as follows. First, critical generators are identified by a new index in the early stage of the time domain simulation. Thus, computational time required to find OMIB(One Machine Infinite Bus) can be reduced significantly. Second, clustering critical machines can be performed even in very stable cases. It enables to be avoid extra calculation of time trajectory that is needed in SIME for classifying the stable cases. Finally, using power-angle trajectory and subdividing contingency classification have improved the screening capability. This algorithm is applied to the fast TSA of the KEPCO system.

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Fabrication of 3D Multilayered Microfluidic Channel Using Fluorinated Ethylene Propylene Nanoparticle Dispersion (불소화 에틸렌 프로필렌 나노 입자 분산액을 이용한 3차원 다층 미세유체 채널 제작)

  • Min, Kyoung-Ik
    • Korean Chemical Engineering Research
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    • v.59 no.4
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    • pp.639-643
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    • 2021
  • In this study, fluorinated ethylene propylene (FEP) nanoparticle as an adhesive for fabricating a three-dimensional multilayered microfluidic device was studied. The formation of evenly distributed FEP nanoparticles layer with 3 ㎛ in thickness on substrates was achieved by simple spin coating of FEP dispersion solution at 1500 rpm for 30 s. It is confirmed that FEP nanoparticles transformed into a hydrophobic thin film after thermal treatment at 300 ℃ for 1 hour, and fabricated polyimide film-based microfluidic device using FEP nanoparticle was endured pressure up to 2250 psi. Finally, a three-dimensional multilayered microfluidic device composed of 16 microreactors, which are difficult to fabricate with conventional photolithography, was successfully realized by simple one-step alignment of FEP coated nine polyimide films. The developed three-dimensional multilayered microfluidic device has the potential to be a powerful tool such as high-throughput screening, mass production, parallelization, and large-scale microfluidic integration for various applications in chemistry and biology.

Mass Screening of Lovastatin High-yielding Mutants through Statistical Optimization of Sporulation Medium and Application of Miniaturized Fungal Cell Cultures (Lovastatin 고생산성 변이주의 신속 선별을 위해 통계적 방법을 적용한 Sporulation 배지 개발 및 Miniature 배양 방법 개발)

  • Ahn, Hyun-Jung;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.297-304
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    • 2007
  • For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

A Study on the Present Status and Future Directions of Maritime Safety Audit (해상교통안전진단제도의 운영현황과 향후 정책방향에 관한 연구)

  • Cho, Ik-Soon
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.17 no.4
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    • pp.399-405
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    • 2011
  • Recently, the navigation risk is increasing significantly with growth of marine traffic volume and construction of marine facilities, water bridges, port development and marine wind farm etc. To reduce this kinds of risk, Ministry of Land, Transport and Maritime Affairs enacted a new law called MSA(Maritime Safety Audit) as a comprehensive maritime traffic safety management scheme in order to ensure safety improvements from the early planning stage to post managing of the development which affect the maritime traffic environment. MSA as a tool for improving maritime traffic safety is a formal safety assessment in the existing or future ship's fairway by an independent audit team. It examines the potential hazards of maritime traffic safety, if necessary, and is to ensure the implementation of appropriate safety measures. The object of this paper is to comprehensively evaluate the achievements and implementation problems of MSA about the 2 years, to define the fundamental problems of MSA by conceptualizing and analyzing MSA limits. MSA requires further examination about the introduction of screening and scoping in order to increase the efficiency and objectivity. It will be required the measures concerning policies directions as a tool for planning process for project owner. It will lead to right understanding concerning audit scheme and used in various ways such as amendments to related law.

Spectrofluorometric Characteristics of the N-Terminal Domain of Riboflavin Synthase (아미노-말단 리보플라빈 생성효소 단백질의 형광 특성)

  • Kim, Ryu-Ryun;Yi, Jeong-Hwan;Nam, Ki-Seok;Ko, Kyung-Won;Lee, Chan-Yong
    • Korean Journal of Microbiology
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    • v.47 no.1
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    • pp.14-21
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    • 2011
  • Riboflavin synthase catalyzes the formation of one molecule of each riboflavin and 5-amino-6-ribitylamino-2,4-pyrimidinedione by the transfer of a 4-carbon moiety between two molecules of the substrates, 6,7-dimetyl-8-ribityllumazine. The most remarkable feature is the sequence similarity between the N-terminal half (1-97) and the C-terminal half domain (99-213). To investigate the structure and fluorescent characteristics of the N-terminal half of riboflavin synthase (N-RS) in Escherichia coli, more than 10 mutant genes coding for the mutated N-terminal domain of riboflavin synthase were generated by polymerase chain reaction. The genes coding for the proteins were inserted into pQE vector designed for easy purification of protein by 6X-His tagging system, expressed, and the proteins were purified. Almost all mutated N-terminal domain of riboflavin synthases bind to 6,7-dimethyl-8-ribityllumazine and riboflavin as fluorescent ligands. However, N-RS C47D and N-RS ET66,67DQ mutant proteins show colorless, indicating that fluorescent ligands were dissociated during purification. In addition, most mutated proteins show low fluorescent intensity comparing to N-RS wild type, whereas N-RS C48S posses stronger fluorescent intensity than that of wild type protein. Based on this result, N-RS C48S can be used as the tool for high throughput screening system for searching for the compound with inhibitory effect for the riboflavin synthase.

The Effects of Cesium, Strontium and Cobalt on Cell Toxicity in the 2D and 3D Cell Culture Platforms (단층 및 입체 세포배양환경에서 세슘, 스트론튬 및 코발트가 세포 독성에 미치는 영향 분석)

  • Kim, Gi Yong;Kang, Sung-Min;Jang, Sung-Chan;Huh, Yun Suk;Roh, Changhyun
    • Korean Journal of Environmental Biology
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    • v.34 no.2
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    • pp.107-115
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    • 2016
  • Currently, there are 442 operating nuclear power plants in the world, and 62 more are under construction. According to this reasoning, the treatment of radioactive waste is important to prevent the environmental ecosystem including humans, animals, and plants. Especially, a leakage of radioactive waste causes not only regional problem but also serious global one. In this study, we demonstrate the effect of radioisotopes (e.g., cesium, strontium, and cobalt) on a 3D culture cell. To develop the 3D cell culture system, we used a 96-well-culture plate with biocompatible agarose hydrogel. Using this method, we can perform the 3D cell culture system with three different cell lines such as HeLa, HepG2, and COS-7. In addition, we conducted a cell viability test in the presence of radioisotopes. Interestingly, the 3D morphological cells showed 42% higher cell viability than those on the 2D against cesium. This result indicates that the 3D platform provides cells morphological and physiological characteristic similar to in vivo grown tissues. Moreover, it overcomes the limitation of conventional cell culture system that can't reflect in vivo systems. Finally, we believe that the proposed approach can be applied a new strategy for simple high-throughput screening and accurate evaluation of metal toxicity assay.