• 제목/요약/키워드: iSC

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Lactobacillus plantarum Bacteriophage SC 921의 phage particle protein 및 genome의 특성 (Phage Particle Proteins and Genomic Characterization of the Lactobacillus plantarum Bacteriophage SC 921.)

  • 김재원;신영재;심영섭;유승구;윤성식
    • 한국미생물·생명공학회지
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    • 제26권2호
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    • pp.117-121
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    • 1998
  • 김치로부터 분리한 Lactobacillus plantarum bacteriophage SC 921은 M.O.I가 0.2일 경우 용균효과가 빠르게 진행됨을 알 수 있었고, SDS-PAGE를 실시하여 phage particle protein을 조사해 본 결과 4개의 major protein으로 구성되어 있는데 이들은 각각 48, 34, 32, 29 kDa으로 구성되어 있다. Exo III로 30분간 반응시킨 후 S1 nuclease를 처리하여 DNA의 형태를 조사해 본 결과 intact DNA는 linear form의 double strand를 유전전달 물질로 가지고 있었다. 제한효소에 대한 절단 효과를 조사한 결과, Sma I에 대해서 1개, Xba I, Cla I, Kpn I, EcoRI에 대해서 각각 2, 4, 5, 6개의 절단부위를 가지고 있으며, Hind III에 대해서는 절단부위가 매우 많은 것을 알 수 있었다. Hind III를 이용하여 intact DNA의 genome size를 측정해본 결과 약 66.5 kbp정도였다. 위의 실험결과와 restriction enzyme mapping을 통해 기존에 알려진 bacteriophage B2와 비교해본 결과 숙주 균주는 같으나 단백질적인 구조나 유전전달물질로 본 구조는 서로 다름을 알 수 있었다.

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Sc 첨가된 Al-Zn-Mg-(Cu)계 알루미늄 합금 압출재의 시효 경화 거동과 기계적 성질 (Age Hardening and Mechanical Property of Extruded Al-Zn-Mg-(Cu) Al Alloys with Sc addition)

  • 심성용;임수근
    • 열처리공학회지
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    • 제20권5호
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    • pp.243-249
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    • 2007
  • The age hardening behavior and mechanical properties of an extruded Al-Zn-Mg-(Cu)-0.1 wt.%Sc alloy were investigated with the Sc addition and ageing temperature. The results showed that the $Al_3Sc$ compounds were formed by Sc addition and distributed preferentially along the extrusion direction. The age hardening of Al-Zn-Mg-Cu-0.1 wt.%Sc alloy which was treated by T6 process was more significant than that of Al-Zn-Mg-0.1 wt.%Sc alloy. The tensile property of Al-Zn-Mg-Cu+0.1 wt.%Sc alloy was also higher than that of Al-Zn-Mg-0.1 wt.%Sc alloy, which is 691 MPa and 584 MPa in strength and 9% and 11% in elongation, respectively.

pET vector를 통한 유전자 재조합 단일사슬 항 B형 림프종 항체의 생산과 면역반응성 평가 (Production of the Recombinant Single Chain Anti-B Cell Lymphoma Antibody and Evaluation of Immunoreactivity)

  • 정재호;최태현;우광선;정위섭;김수관;천기정;최창운;임상무
    • Nuclear Medicine and Molecular Imaging
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    • 제40권4호
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    • pp.211-217
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    • 2006
  • 재조합된 scFv lym-1의 세포결합 실험에서 높은 면역반응성을 보였으며, 비특이 반응에서 모항체인 IgG lym-1에 의해 결합이 억제됨을 확인하였고, 동물 영상을 통해 IgG보다 분자랑이 작은 scFv lym-1 항체가 빠른 체내대사율과 종양섭취율을 보여 방사면역진단에 유용할 것으로 보여진다.

마우스 대식세포주인 RAW 264.7 세포에서 오공(蜈蚣)의 항염증 효과 (Anti-Inflammatory Effect of Aqueous Extract of Scolopendrae Corpus in RAW 264.7 Cells)

  • 조일주;최미옥;박민철;송호준;박성주
    • 대한본초학회지
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    • 제26권3호
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    • pp.23-29
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    • 2011
  • Objective : The purpose of this study was to investigate the anti-inflammatory effects of aqueous extract from Scolopendrae Corpus (SC) on lipopolysaccharide (LPS)-induced inflammatory response. Methods : To evaluate the anti-inflammatory effects of SC, we examined the inflammatory mediators such as nitric oxide (NO) and pro-inflammatory cytokines (TNF-a, inteleukin (IL)-$1{\beta}$ and IL-6) on RAW 264.7 cells. We also examined molecular mechanisms such as mitogen-activated protein kinases (MAPKs) and inhibitory kappa B a ($I{\kappa}$-Ba) using western blot. Furthermore, we also investigated the effect of SC on LPS-induced endotoxin shock. Results : Extract from SC itself had not any cytotoxic effect in RAW 264.7 cells. Aqueous extract from SC inhibited LPS-induced NO production and iNOS expression. SC pre-treatment also inhibited IL-$1{\beta}$, IL-6 production in RAW 264.7 cells. To investigate inhibitory effects of SC on inflammatory mediators, activation of MAPKs was examined. SC inhibited the phosphorylation of p38 kinases (p38), c-Jun $NH_2$-terminal kinase (JNK) and also the degradation of $I{\kappa}$-$B{\alpha}$ in RAW 264.7 cells stimulated with LPS. Furthermore, SC administration reduced LPS-induced endotoxin shock. Conclusion : SC down-regulated LPS-induced production of inflammatory mediators through inhibition of activation of p38, JNK and degradation of $I{\kappa}$-$B{\alpha}$. Taken together, our results suggest that SC may be a beneficial drug against inflammatory diseases such as sepsis.

경사냉각판을 이용한 Al-Zn-Mg-(Sc) 반응고 합금 제조의 최적화 및 재가열 특성 (Optimum Fabrication Conditions and Reheating Characteristic of Semi-Solid Al-Zn-Mg-(Sc) Alloy by Inclined Cooling Plate)

  • 김태훈;심성용;박형원;임수근
    • 한국주조공학회지
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    • 제29권5호
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    • pp.213-219
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    • 2009
  • Optimum conditions for production of semi-solid Al-Zn-Mg alloy billets was carried out by the Taguchi design method. And, Al-Zn-Mg alloy billets contained Sc (free, 0.1 and 0.3 mass %) were fabricated at optimum conditions. Evolution of microstructure in semi-solid state was investigated through various liquid fractions, holding times and holding temperatures. The Al-Zn-Mg alloy billets reheated at $615^{\circ}C$ during 30min are grain growth and it was fractured due to increasing liquid fraction before quenching. And, during reheating up to $600^{\circ}C$, grain growth of Al-Zn-Mg alloy billets contained Sc (0.1 and 0.3 mass %) was not occurred in comparison with those of Al-Zn-Mg alloy without Sc. It was thought that $Al_3Sc$ phases have a pinning effect in grain boundary and Sc content of 0.1 mass% is able to inhibit grain growth effectively through reheating process.

효모 발현 시스템을 이용 PAP(Pokeweed Antiviral Protein) 유전자의 세포독성 연구 (Cytotoxicity Test of Pokeweed Antiviral Protein Type I Gene by Using Yeast Expression System)

  • 김선원;박성원;강신웅;이영기;이종철;최순용;이청호
    • 한국연초학회지
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    • 제23권2호
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    • pp.133-140
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    • 2001
  • PAP-I cDNA was synthesized from total RNA of Phytolacca americana leaves by RT-PCR, and then subcloned to recombinant vector pBluescript II SK-. Using PCR with primers designed in our laboratory, we could get the 9 deletion mutant PAP-I cDNA fragments. The first of the fragments was deleted by 66bp from immature N-terminal and then the rest were deleted by 90bp sequentially. Sequentially deletion mutant PAP-I cDNAs were inserted to pAc55M, on down-stream of gall promoter. Recombinant pAc55M was transformed to yeast cells, psy1 and the cells were spreaded on SC_urn-/glucose plate media. Colonies on SC_ura-/glucose plate were streaked on the same position of SC_ura-/glucose and SC_ura-/galactose plate, and we selected colonies growing on both plates, which carry non-cytotoxic deleted mutant PAP-I cDNA. We selected 4 deletion mutant PAP-I cDNAs which have not cytotoxicity.

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꽃송이버섯 추출물이 RAW 264.7 세포에서 TNF-α, iNOS, IL-1β 유전자 발현에 미치는 영향 (Effect of Water Extract of Sparasis crispa on the Expression of TNF-α, iNOS and IL-1β Genes in RAW 264.7 Cells)

  • 한효상
    • 대한통합의학회지
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    • 제9권1호
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    • pp.163-171
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    • 2021
  • Purpose : The purpose of this study was to examine the anti-inflammatory effects of Sparassis crispa (SC). SC is a well-known traditional herbal remedy and its mushroom is used for treatment of inflammation. Many diseases that are increasing recently have characteristics of inflammatory diseases. Researchers are finding bioactive substances from natural products that can promote treatment and prevention of inflammation. We investigated the effect of water extracted from SC on the expression of effector genes involved in the function of RAW 264.7 cells. Methods : Effects of RAW 264.7 cells on cell viability, antioxidation, and mRNA expression were examined using water extracts from SC. A 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed to determine the effect of water extracts from SC on cell viability in RAW 264.7 cells. Inflammation of RAW 264.7 cells induced by lipopolysaccharide (LPS) treatment and expression levels of inflammatory cytokine TNF-α, iNOS and IL-1β gene were analyzed using quantitative reverse transcription PCR (qRT-PCR) analysis. Results : The MTS assay was performed on RAW 264.7 cells after treatment with various concentrations of water extracts of SC. Treatment of RAW 264.7 cells with water extracts from SC and LPS at a concentration of 0.125, 0.5 mg/㎖ for twenty four hours promoted mRNA expression of TNF-α, iNOS and IL-1β. Conclusion : MTS assay was applied to RAW 264.7 cells after various concentrations of water extracts of SC. Through experimental demonstration of anti-oxidant and anti-inflammatory effects of water extracts from SC, we suggest that SC is a valuable material for the prevention and treatment of various inflammatory diseases.

형광 물질 직접 표지를 위한 Poly Lysine 도입 Lym-1 단일사슬 항체의 제조 및 면역반응성 평가 (Production and Evaluation of Immunoreactivity of Poly Lysine-Tagged Single Chain Fragment Variable (ScFv) Lym-1 Antibody for Direct Conjugation to Fluorescence Dye)

  • 정재호;최태현;우광선;정위섭;강주현;정수영;최창운;임상무;천기정
    • Nuclear Medicine and Molecular Imaging
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    • 제43권5호
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    • pp.487-494
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    • 2009
  • 목적: 작은 크기의 재조합 단일사슬 항체는 빠른 혈중 제거율과 종양의 항체 집적율이 증가되는 등의 장점을 가지고 있다. 반면에 항체의 작은 크기는 방사성 또는 형광물질의 표지를 위한 킬레이터 결합에 중요한 아미노산 그룹의 감소를 의미하기도 한다. 본 연구에서는 단일사슬 lym-1 염기서열 C-말단에 lysine 아미노산 태그를 삽입하여 형광 물질의 직접표지 및 그 표지수율 증가를 확인하고자 하였다. 대상 및 방법: 대장균 pET-22b (+) 벡터에 재조합 된 lysine 삽입 단일사슬 lym-1유전자는 대장균 BL21 (DE3)에 형질전환하여 발현하였다. 생산된 lysine lym-1 항체는 Ni-NTA 컬럽과 분자량 컬럼을 사용해 정제하였고. 단백질 전기 영동과 western blot을 통해 확인하였다. lysine lym-1 항체에 방사성 동위원소인 I-124, I-125, I-131 과 Tc-99m를 표지하여 그 수율을 확인하였으며 유세포계측기를 사용해 형광물질인 FITC가 직접표지된 라이신 lym-1 항체의 면역반응성을 사람의 버킷 림프종 세포주인 Raji 세포주에서 면역반응성을 확인하였다. 결과 Lysine도입 단일사슬 lym-1 항체는 두 과정의 정제를 통하여 획득하였으며 그 크기는 약 48 KDa이었고, 방사성동위원소인 I-124, I-125, I-131과 Tc-99m의 표지수율은 각각 >99%, >99%, >95%, >99%로 확인되었다. 유세포계측을 통한 lysine 도입 단일사슬 lym-1항체의 면역반응성은 기존의 단일사슬 lym-1항체와 유사함을 확인하였다. 결론: 재조합 lym-1 항체에 형광 물질을 직접 표지하기 위한 lysine 아미노산의 도입은 항체의 면역반응성 감소를 최소화 시키면서 직접표지 수율을 증가시킬 수 있는 유용한 방법임을 확인하였다.

화분분석을 이용한 아산시 온양천 유역의 후빙기 후기 환경변화 (A Pollen Analysis on the Environmental Changes during the Later Half of the Postglacial Age around the Basin of Onyang River, Asan)

  • 박지훈
    • 한국지형학회지
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    • 제17권1호
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    • pp.39-48
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    • 2010
  • 충남 온양천 유역의 후빙기 환경변화를 밝히기 위한 사례연구로서 온양천의 지류인 금곡천의 상류부(이하 조사지역)에 분포하는 선상지 퇴적층을 대상으로 화분분석을 실시하였다. 시료채취지점은 해발고도 약 67.5m로 현재 냉온대 중부 식생대에 속한다. 분석결과는 다음과 같다. 조사지역 일대는 약 3,000yrB.P. 이후 크게 SC-I기(期)(Pinus가 우점하는 침엽수림시대) → SC-II기(期)(Quercus·Castanea가 우점하는 낙엽활엽수림시대) → ③ SC-III기(期)(Pinus-Quercus-Ulmus/Zelkova가 우점하는 침활혼효림시대)로 변천했다. 각 시대의 연대는 SC-I기(期)와 SC-II기(期)가 약 3,000~2,000yrB.P., SC-III기(期)가 약 2,000yrB.P.~현재로 수렴된다. 전국적 후빙기 화분대와 대비하면, SC-I기(期)와 SC-II기(期)는 RIIIa 그리고 SC-III기(期)는 RIIIb에 대비된다. 그리고 약 2,000yrB.P. 이후 조사지역 일대는 삼림파괴에 따른 소나무림의 분포역이 확대되었으며, 또한 메밀화분의 출현율이 높은 점 등을 고려할 때, SC-III기(期)는 인간간섭의 시대로서 이 시대에 들어서면서부터 삼림파괴가 본격화되었고, 당시 거주민들은 농경생활을 했었을 것으로 추정된다.

Double-stranded RNA Induces Inflammatory Gene Expression in Schwann Cells: Implication in the Wallerian Degeneration

  • Lee, Hyun-Kyoung;Park, Chan-Hee;Choi, Se-Young;Oh, Seog-Bae;Park, Kyung-Pyo;Kim, Joong-Soo;Lee, Sung-Joong
    • The Korean Journal of Physiology and Pharmacology
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    • 제8권5호
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    • pp.253-257
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    • 2004
  • Schwann cells play an important role in peripheral nerve regeneration. Upon neuronal injury, activated Schwann cells clean up the myelin debris by phagocytosis, and promote neuronal survival and axon outgrowth by secreting various neurotrophic factors. However, it is unclear how the nerve injury induces Schwann cell activation. Recently, it was reported that certain cytoplasmic molecules, which are secreted by cells undergoing necrotic cell death, induce immune cell activation via the toll-like receptors (TLRs). This suggests that the TLRs expressed on Schwann cells may recognize nerve damage by binding to the endogenous ligands secreted by the damaged nerve, thereby inducing Schwann cell activation. Accordingly, this study was undertaken to examine the expression and the function of the TLRs on primary Schwann cells and iSC, a rat Schwann cell line. The transcripts of TLR2, 3, 4, and 9 were detected on the primary Schwann cells as well as on iSC. The stimulation of iSC with poly (I : C), a synthetic ligand for the TLR3, induced the expression of $TNF-{\alpha}$ and RANTES. In addition, poly (I : C) stimulation induced the iNOS expression and nitric oxide secretion in iSC. These results suggest that the TLRs may be involved in the inflammatory activation of Schwann cells, which is observed during Wallerian degeneration after a peripheral nerve injury.