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김치에서 산내성을 가진 Leuconostoc mesenteroides LM3의 분리
사금희,백상규,윤혜선,강경희,정진국,김일섭,문혜연,김사열,유춘발 한국생명과학회 2002 생명과학회지 Vol.12 No.6
In order to understand stress response of Leuconostoc mesenteroides against lactic acid, a new Leuconostoc sp. which has acid tolerance was isolated from various Kimchi samples. And it identified as Leuconostoc mesenteroides LM3 by comparing its fatty acid composition with reference strain. Its growth pattern was investigated by adding a given concentration of lactic acid at the lag phase to the stationary phase. In the DeMan, Rogosa, Sharpe (MRS) media containing over 0.4% (final v/v) lactic acid, this strain slowed slowly After exposure of the stationary phase cells to 4% of lactic acid for 60 min, this strain could survive, whereas a reference strain, Leuconostoc mesenteroides KCTC3505, showed no survival. And changes of trehalose concentration, the activity of trehalase and ATPase in the growth of Leuconostoc mesenteroides LM3 after addition of 0.6% (final v/v) lactic acid were investigated : After exposure to lartic acid, trehalose concentration in this strain was increased in comparison with no treatment, but its trehalase activity was not changed. And its ATPase activity was constant, and intracellular pH was almost constant. This result meant Leuconostoc mesenteroides LM3 should have a tolerance against lactic acid. It remains to further study the mechanism of this acid tolerance.
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βig-h3 절편-RGD 재조합단백의 만성염증성 관절염 치료효과
장지애 ( Ji Ae Jang ),강진희 ( Jin Hee Kang ),사금희 ( Keum Hee Sa ),한승우 ( Seung Woo Han ),서재석 ( Jae Seok Seo ),김경훈 ( Kyung Hoon Kim ),남언정 ( Eon Jeong Nam ),김인산 ( In San Kim ),강영모 ( Young Mo Kang ) 대한류마티스학회 2012 대한류마티스학회지 Vol.19 No.2
Objective. βig-h3 is a 68kDa extracellular matrix protein which is overexpressed in synovial tissues of rheumatoid arthritis (RA). Previous results proved that βig-h3 fragments are relevant to adhesion and migration of synovial fibroblast and angiogenesis through interaction with αvβ 3 integrin. We designed a recombinant βig-h3 protein consisting of a fas-1 domain and RGD motif and evaluated the therapeutic efficacy in RA. Methods. Inhibitory effect of adhesion and migration of NIH3T3 cell line was evaluated in 96 well microtiter and transwell plates coated with βig-h3. Clinical arthritis index was evaluated after treating CIA mice with MFK12. Immunohistochemical staining in synovial tissues were performed. Expression of transcripts and proteins of inflammatory mediators were analyzed by semi-quantitative RT-PCR and immunoblotting. Results. Recombinant protein consisted of 4th fas-1 domain truncated for H1 and H2 sequences and RGD peptide (MFK12), had M.W. of 10.4kDa. βig-h3 mediated adhesion and migration of NIH3T3 cell line were significantly inhibited in a dose-dependent manner. Arthritis severity and incidence were efficiently reduced when CIA mice were treated with MFK12 at 30 mg/kg/day compared with the control. Immunohistochemical staining of joint tissues in MFK12 treated mice exhibited reduced angiogenesis. In treated mice, expression of transcripts regarding inflammatory mediators was markedly suppressed and immunoblotting of ICAM-1 and RANKL from whole extract of hind paws also showed a significant reduction. Conclusion. This study shows that MFK12 is effective in treating RA, although further study is warranted to improve the therapeutic efficacy. Key Words. Rheumatoid arthritis, Inflammation, βig-h3, collagen-induced arthritis, Fas-1
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류마티스 활막염에 있어 염증매개물질에 의한 Transforming Growth Factor-β-inducible Gene-h3 (βig-h3) 생산 조절 기전
강영모 ( Young Mo Kang ),김성일 ( Sung Il Kim ),김정섭 ( Jeong Seup Kim ),유동완 ( Dong Wan You ),사금희 ( Kheum Hee Sa ),박은주 ( Eun Ju Park ),김성욱 ( Sung Uk Kim ),서재석 ( Jae Seok Seo ),한승우 ( Seung Woo Han ),남언정 ( Eon 대한류마티스학회 2005 대한류마티스학회지 Vol.12 No.2
Objective: To investigate the expression pattern of transforming growth factor-β-inducible gene-h3 (βig-h3) within rheumatoid synovial tissue and the regulation of βig-h3 synthesis in fibroblast-like synoviocyte (FLS). Methods: Synovial tissues obtained from patients with rheumatoid arthritis and osteoarthritis were obtained during joint replacement surgery. βig-h3 expression was evaluated with immunohistochemical stain. FLS was isolated from synovial tissues and stimulated with cytokines including TGF-β, TNF-α, IL-1β, IFN-γ, IL-6, IL-4, and IL-10. βig-h3 synthesis was measured using semiquantitative RT-PCR, ELISA, immunofluorescence stain, and flow cytometry. Results: Expression of βig-h3 was diffuse and abundant in both lining and sublining layers of rheumatoid synovium, which was more prominent than those of osteoarthritis. Production of βig-h3 in FLS was regulated by TGF-β1 in a dose-dependent manner and was highest at 5 ng/mL of TGF-β1. TNF-α and IL-1β upregulated the production of βig-h3 from FLS synergistically with TGF-β1 but other cytokines such as IL-4, IL-6, IL-10 did not affect. βig-h3 synthesis was efficiently inhibited by dexamethasone at higher dose (100 nM) but not by cyclosporine-A. Conclusion: Production of βig-h3, which is highly upregulated in rheumatoid synovitis, is differentially regulated by inflammatory cytokines.
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류마티스관절염에서 TGF-β-inducible Gene-h3 (βig-h3)에 의한 활막세포 부착기전
조현정 ( Hyung Jung Cho ),박재용 ( Jae Yong Park ),경희수 ( Hee Soo Kyung ),김인산 ( In San Kim ),강영모 ( Young Mo Kang ),남언정 ( Eon Jeong Nam ),송은주 ( Eun Joo Song ),김지민 ( Ji Min Kim ),서재석 ( Jae Seok Seo ),사금희 ( Keu 대한류마티스학회 2008 대한류마티스학회지 Vol.15 No.3
Objective: βig-h3 is an extracellular matrix protein, which is overexpressed in synovial tissues of rheumatoid arthritis (RA) similar to adhesive glycoproteins. We sought to evaluate the compensatory role of βig-h3 with adhesive glycoproteins in mediating the adhesion of fibroblast-like synoviocytes (FLS) and to confirm the inhibitory effect of YH18 peptide of the 2nd fas-1 domain in βig-h3-mediated adhesion. Methods: The adhesion of FLS isolated from synovial tissues of RA, was evaluated in 96 well microtiter plate coated with matrix proteins. Inhibitory effect of YH18 peptides from the 2nd and 4th fas-1 domains was estimated in βig-h3-mediated adhesion of FLS. Results: The adhesion of FLS on βig-h3 was weaker than that of fibronectin and vitronectin. The βig-h3-mediated adhesion was enhanced by the stimulation with phorbol myristate acetate (PMA), but not by cytokines and growth factors. Combination of fibronectin with βig-h3 synergistically enhanced the adhesion of FLS, in contrast to the additive effect of vitronectin combined with βig-h3. YH18 peptide of the 2nd fas-1 domain did not block the βig-h3-mediated adhesion of FLS. Conclusion: Our results reveal that βig-h3 may regulate the adhesion of FLS through the interaction with adhesive glycoproteins and confirm that the essential motifs mediating adhesion on βig-h3 are different according to the type of cells.
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