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Development of Hepatitis C Virus (HCV) Genome-Targeting Hammerhead Ribozyme Which Activity Can Be Allosterically Regulated by HCV NS5B RNA Replicase (C형 간염바이러스(HCV)의 NS5B RNA Replicase에 의해 그 활성이 조절되는 HCV지놈 표적 Hammerhead 리보자임 개발)

  • Lee, Chang-Ho;Lee, Seong-Wook
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.159-165
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    • 2007
  • For the development of basic genetic materials for specific and effective therapeutic approach to suppress multiplication of hepatitis C virus (HCV), HCV internal ribosome entry site (IRES)-targeting hammerhead ribozyme which activity is allosterically regulated by HCV regulatory protein, NS5B RNA replicase, was developed. The ribozyme targeted most effectively to +382 nucleotide (nt) site of HCV IRES RNA. The allosteric ribozyme was designed to be composed of sequence of RNA aptamer to HCV NS5B, communication module sequence which can transfer structural transition for inducing ribozyme activity upon binding NS5B to the aptamer, and sequence of ribozyme targeting +382 nt of HCV IRES. Noticeably, we employed in vitro selection technology to identify the most appropriate communication module sequence which can induce ribozyme activity depending on the US5B protein. We demonstrated that the ribozyme was nonfunctional either in the absence of any proteins or in the presence of control bovine serum albumin. In sharp contrast, the allosteric ribozyme can induce activity of cleavage reaction with HCV IRES RNA in the presence of the HCV NS5B protein. This allosteric ribozyme can be used as lead compound for specific and effective anti-HCV agent, tool for highthroughput screening to isolate lead chemicals for HCV therapeutics, and ligand for biosensor system for HCV diagnosis.

A Exploratory Comparison Study on Anchoring Effect according to Regularity of Snack's Shape (스낵모양의 규칙성에 따른 앵커링 효과의 탐색적 비교연구)

  • Oh, Ji-Won;Lee, Hyun-Kyung;Kim, Gwi-Gon
    • Journal of Digital Convergence
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    • v.14 no.6
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    • pp.237-244
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    • 2016
  • The purpose of this study examines the influence of anchoring and adjustment according to regularity of snack's shape. Especially this study choose 'choco chip' as a snack with regularity to compare with 2012's study. The results of this study are as follows. 1) The anchoring and adjustment also appears in 'choco chip' with regularity. But statistical meaning is different from the result of prior study. 2) The difference of anchoring effect reduced according to additional verbal information in 'choco chip' with regularity. but has no statistical meaning of interaction effect. However the causes of two results are different. 3) The interaction effect of cognitive load(without vs. moderate vs. high) is meaningful statistically. difference of anchoring effect reduced according to additional verbal information in 'choco chip' with regularity in 'choco chip' with regularity. This study provides strategic implication that it overcomes the limitation of the prior resuarch and may reinforce the biased judgement by cognitive overload.

Evaluation of Potential for Antioxidant and Anti-inflammatory Material of Schisandra chinensis and Cudrania tricuspidata of Cultivated in Sunchang-gun (순창군 재배 오미자와 꾸지뽕의 항산화 및 항염증 소재 활용 가능성 평가)

  • Jeong-Ho Lee;Gyeong-Ok Jeong;Kwang-Hyeon Moon;Se-Won Lee;Seong-Hyeon Lee;Gwang-Min Lee;Yeo-Jin Yoo;Eui-Yong Lee;Hyun-Jin Tae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.08a
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    • pp.82-82
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    • 2020
  • 염증 (Inflammation)은 물리적인 상처나 세균감염이 되었을 때 손상된 조직을 재생하고 신체를 방어하기 위해 일어나는 선천성 면역반응으로 알려져 있다. 주로 선천면역을 담당하는 대식세포는 lipopolysaccharide, reactive oxygen species와 cytokine 등에 의해 활성화되어 tumor necrosis factor-α. interleukin-1β, interleukin-6 등 염증인자들의 생성에 관여한다. 특히, 산화질소는 superoxide 음이온과 쉽게 반응하고 peroxynitrite와 같은 독성이 강한 산화제를 생성하여 단백질 및 지질의 과산화를 유도하고 세포독성을 일으키는 것으로 보고되고 있다. 따라서 본 연구에서는 항산화 및 항염증 소재를 탐색하기 위해 오미자 (Schisandra chinensis), 꾸지뽕 (Cudrania tricuspidata)을 이용하여 항산화 활성을 평가하고자 하였으며, 세포주를 활용한 세포독성 및 항염증 활성을 확인하고자 하였다. 본 연구는 전북 순창군에서 재배된 오미자, 꾸지뽕을 열 건조(60℃) 통해 건조한 후 분말화하였다. 최적 추출 조건 선정을 위해 다양한 용매 (열수, 증류수, 주정 20, 40, 60%), 온도 (25, 40, 60, 80℃) 및 시간 (1, 2, 3, 4, 5 h) 조건에서 추출된 추출물의 총 폴리페놀 함량을 비교함으로써 최적 조건을 선정하였다. 오미자와 꾸지뽕의 DPPH 및 ABTS radical 소거 활성, 총 플라보노이드 함량을 확인하여 항산화능 및 기능성 성분 함량을 평가하였다. 또한 대식세포주인 Raw 264.7을 활용하여 MTT assay, 산화질소 생성 억제 활성을 확인하여 세포독성 및 항염증 활성을 평가하였다. 실험 결과, 오미자 및 꾸지뽕은 각각 주정 40%, 60℃ 그리고 증류수, 60℃에서 추출 시 가장 높은 총 폴리페놀 함량 (약 98.3 mg GAE/g 및 88.2 mg GAE/g)을 함유하며, DPPH 라디칼 소거 활성은 약 44.6% 및 24.4%, ABTS 라디칼 소거 활성은 약 30.3% 및 40.8%로 확인되었다. 총 플라보노이드 함량은 약 21.80 mg QE/g 및 35.68 mg QE/g으로 확인되었다. 또한 오미자 및 꾸지뽕 기능성 추출물을 100 ug/mL 처리 시 세포 독성이 나타나지 않는 것으로 확인되었으며, NO 생성량을 약 56.3% 및 21.7% 저감시켜 항염증 효능을 나타내는 것으로 확인되었다.

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Screening of Effective Medium Composition for the Cultivation of Lactobacillus plantarum and Lactobacillus reuteri Using Statistical Methods (통계적 방법을 이용한 Lactobacillus plantarum과 Lactobacillus reuteri 의 유효 배지 성분의 탐색)

  • Kim, Dong-Woon;Cho, Sang-Buem;Kim, Young-Hwa;Lee, Sung-Daw;Jung, Hyun-Jung;Kim, Sang-Ho;Cho, Kyu-Ho;Sa, Soo-Jin;Kim, In-Cheul;Won, Mi-Young;Kim, Su-Ok;Kim, Soo-Ki
    • Journal of Life Science
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    • v.22 no.5
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    • pp.575-581
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    • 2012
  • This study was conducted to develop an economical optimum medium composition for the mass production of $Lactobacillus$ $plantarum$ and $Lactobacillus$ $reuteri$, livestock probiotics. Medium ingredient factors were selected on the basis of MRS broth composition, and the 15 ingredient variables were as follows: sucrose, glucose, molasses, yeast extract, corn steep liquor, soy peptone, dipotassium phosphate, manganese chloride, magnesium chloride, tween 80, sodium chloride, sodium acetate, ammonium citrate, sodium sulphate, and ferrous sulphate. The Plackett Burman design, consisting of 20 runs, was employed for the analysis of ingredient effects on cell growth of $L.$ $plantarum$ and $L.$ $reuteri$. As a result, sucrose, glucose, molasses, yeast extract, corn steep liquor, soy peptone, sodium acetate, and ammonium citrate positively influenced the growth of $L.$ $plantarum$. Additionally, yeast extract, soy peptone, $K_2PHO_4$, and tween 80 positively influenced the growth of $L.$ $reuteri$. Positive effects were found from sucrose, yeast extract, and soy peptone in the integrated analysis of the effects of both $L.$ $plantarum$ and $L.$ $reuteri$. Finally, effective medium components for both strains were found as follows: sucrose (20.0 g/l), glucose (5.0 g/l), soy peptone (11.0 g/l), yeast extract (5.0 g/l), $K_2PHO_4$ (0.2 g/l), $CH_3COONa$ (2 g/l), and $MgCl_2$ (0.02 g/l).