• Title/Summary/Keyword: iSC

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

  • 김재원;신영재;심영섭;유승구;윤성식
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.117-121
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    • 1998
  • Bacteriophage SC 921 of Lactobacillus plantarum, isolated from kimchi, showed high lytic effects at 0.2 M.O.I. level. The phage particle contained 4 major proteins (48, 34, 32, 29 kDa). Intact DNA of phage SC 921 is a double stranded linear molecule, and the genomic size is approximately 66.5 kilobase pairs (kbp). Restriction analysis of the genome showed that Sma I gave single site cut and Xba I gave 2 site cuts, while Cla I, Kpn I, and EcoR I formed 4, 5, and 6 cuts, respectively. Hind III digested phage DNA to many fragments. A restriction map of genomic DNA was constructed using the restriction endonuclease Kpn I, Sma I, and Xba I. Bacteriophage SC 921 was compared with B2 phage which had been reported to infect Lactobacillus plantarum ATCC 8014(KCCM l1322). Bacteriophage SC 921 differs from B2 phage at least in thr size of its genome and phage particle proteins.

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

  • Shim, Sung Yong;Lim, Su Gun
    • Journal of the Korean Society for Heat Treatment
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    • v.20 no.5
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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.

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

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kang-Sun;Chung, Wee-Sup;Kim, Soo-Gwan;Cheon, Gi-Jeong;Choi, Chang-Woon;Lim, Sang-Moo
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.4
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    • pp.211-217
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    • 2006
  • Purpose: Recombinant ScFv lym-1 was produced, using pET vector system for large scale production. Methods: ScFv lym-1 gene inserted pET-22b (+) vector, was expressed in E.coli BL-21 strain. ScFv lym-1 antibody extracted from periplasm, was purified with His-Taq column. To evaluated immunoreactivity with Raji cell, ScFv lym-1 was labeled with I-125 and I-125 ScFv lym-1 was purified with desalting column. Raji cell was injected into the C57BR/cdJ SCID mice. Gamma camera imaging were taken time point at 1, 8, 24, and 48 hr with 8 mm pinhole collimator. Results: An active scFv lym-1 could be produced in E. coli with soluble iron using PET vector system. Immuuoreaetivity and affinity constant of IgG lym-1 were 54% and $1.83{\times}10^9M^{-1}$, respectively, and those of scFv lym-1 were 53.7% and $1.46{\times}10^9M^{-1}$, respectively. Biodistribution of I-125 scFv lym-1 antibody showed faster clearance in blood, spleen, kidney and than I-125 IgG lym-1 antibody. Gamma camera image of I-125 scFv lym-1 antibody showed faster clearance and tumor targeting liver than I-125 IgG lym-1 antibody. Conclusions: In vitro properties of scFv lym-1 were similar to those of IgG lym-1. ScFv lym-1 showed faster blood clearance than IgG lym-1 There results suggest that scFv lym-1 antibody can be useful for tumor imaging agent.

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

  • Jo, Il-Joo;Choi, Mee-Ok;Park, Min-Cheol;Song, Ho-Joon;Park, Sung-Joo
    • The Korea Journal of Herbology
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    • v.26 no.3
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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.

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

  • Kim, Tae-Hun;Shim, Sung-Yong;Park, Hyung-Won;Lim, Su-Gun
    • Journal of Korea Foundry Society
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    • v.29 no.5
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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.

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

  • 김선원;박성원;강신웅;이영기;이종철;최순용;이청호
    • Journal of the Korean Society of Tobacco Science
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    • v.23 no.2
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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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Effect of Water Extract of Sparasis crispa on the Expression of TNF-α, iNOS and IL-1β Genes in RAW 264.7 Cells (꽃송이버섯 추출물이 RAW 264.7 세포에서 TNF-α, iNOS, IL-1β 유전자 발현에 미치는 영향)

  • Han, Hyo-Sang
    • Journal of The Korean Society of Integrative Medicine
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    • v.9 no.1
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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.

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

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kwang-Sun;Chung, Wee-Sup;Kang, Joo-Hyun;Jeong, Su-Young;Choi, Chang-Woon;Lim, Sang-Moo;Cheon, Gi-Jeong
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.487-494
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    • 2009
  • Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

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

  • PARK, Ji-Hoon
    • Journal of The Geomorphological Association of Korea
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    • v.17 no.1
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    • pp.39-48
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    • 2010
  • This is a case study to research the environmental changes that occurred during the Latter Half of the postglacial age around the Basin of Onyang River in Asan, Korea. In line with this purpose, the author performed a pollen analysis and a radiocarbon dating on the deposits of alluvial fan around the upper Geumgok River, a tributary of Onyang River. Sampling point was at the altitude of about 67.5 meters, which belongs to the central zone of the cool temperate forest. The followings are the results of the study. The study area has passed through SC-I (the coniferous forest period in which Pinus was dominant), SC-II (the deciduous broad-leaved forest period in which Quercus and Castanea were dominant) and SC-III (the mixed conifer and deciduous broad-leaved forest period, in which Pinus, Quercus and Ulmus/Zelkova were dominant) respectively since about 3,000 yrB.P. SC-I period and SC-II period are presumed to be between about 3,000 and 2,000 yrB. P., and SC-III period to begin after 2,000 yrB.P. In comparison with the nationwide pollen zone during the postglacial age, SC-I and SC-II periods are contrasted with the R-IIIa zone and also the SC-III zone with the RIIIb zone. In addition, it is assumed that Pinus densiflora forest luxuriated there since 2,000 yrB.P. due to the destruction of forests, and that a lot of Fagopyrum pollen appeared; altogether, it was the so-called human interference period, from which forests began to be markedly destroyed. It is concluded that in those days inhabitants leaded agricultural life.

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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    • v.8 no.5
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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.