• Title/Summary/Keyword: Alpha 4 beta 7

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Cloning of $\beta$-Glucosidase Gene from Streptomyces coelicolor A3(2) and Characterization of the Recombinant $\beta$-Glucosidase Expressed in Escherichia coli (Streptomyces coelicolor A3(2)로 부터 $\beta$-Glucosidase 유전자 클로닝 및 재조합 효소의 특성)

  • Kim, Jae-Young;Kim, Bong-Kyu;Yi, Yong-Sub;Kang, Chang-Soo;Ahn, Joong-Hoon;Lim, Yoong-Ho
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.99-104
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    • 2009
  • The $\beta$-glucosidase gene from Streptomyces coelicolor A3(2) was cloned and expressed in Escherichia coli. The ORF consisted of 1377 nucleotides encoding 51 kDa in a predicted molecular weight. Effects of pH indicated that the $\beta$-glucosidase showed similar activity using $\alpha$-pNPG($\rho$-nitrophenyl-$\alpha$-D-glucopyranoside), $\beta$-pNPG($\rho$-nitrophenyl-$\beta$-D-glucopyranoside), and $\beta$-pNPF($\rho$-nitrophenyl-$\beta$-D-fucopyranoside) at range of pH 3 to 10, and high activity using $\beta$-pNPGA ($\rho$-nitrophenyl-$\beta$-D-galactopyranoside) from pH 5 to 10, especially, 3.3 times higher activity at pH 9. Effects of temperature indicated that the $\beta$-glucosidase showed low activity using $\alpha$-pNPG, $\beta$-pNPG, and $\beta$-pNPF from $20^{\circ}C$ to $70^{\circ}C$, and increased activity using $\beta$-pNPGA from $30^{\circ}C$ to $50^{\circ}C$, 1.8 times higher activity at $50^{\circ}C$ than at $30^{\circ}C$. According to activity determination of other substrates, the enzyme was active on daidzin, genistin, and glycitin, inactive on esculin and apigenin-7-glucose. The EDTA and DTT as reducing agents inhibited $\beta$-glucosidase activity, but SDS and mercaptoethanol did not inhibit. Monovalent or divalent metal ions such as $MnSO_4$, $CaCl_2$, KCl, and $MgSO_4$ did not inhibited $\beta$-glucosidase activity. $CuSO_4$ and NaCl showed low inhibition, and $ZnSO_4$ inhibited 3.3 times higher than control.

Steroids from the Cold Water Starfish Ctenodiscus crispatus with Cytotoxic and Apoptotic Effects on Human Hepatocellular Carcinoma and Glioblastoma Cells

  • Quang, Tran Hong;Lee, Dong-Sung;Han, Se Jong;Kim, Il Chan;Yim, Joung Han;Kim, Youn-Chul;Oh, Hyuncheol
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2335-2341
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    • 2014
  • Chemical investigation on the methanol extract of the starfish Ctenodiscus crispatus resulted in the isolation of five steroids, (22E,$24{\zeta}$)-26,27-bisnor-24-methyl-$5{\alpha}$-cholest-22-en-$3{\beta}$,5,$6{\beta}$,$15{\alpha}$,25-pentol 25-O-sulfate (1), (22E,24R,25R)-24-methyl-$5{\alpha}$-cholest-22-en-$3{\beta}$,5,$6{\beta}$,$15{\alpha}$,25,26-hexol 26-O-sulfate (2), (28R)-24-ethyl-$5{\alpha}$-cholesta-$3{\beta}$,5,$6{\beta}$,8,$15{\alpha}$,28,29-heptaol-24-sulfate (3), (25S)-$5{\alpha}$-cholestane-$3{\beta}$,5,$6{\beta}$,$15{\alpha}$,$16{\beta}$,26-hexaol (4), and ${\Delta}7$-sitosterol (5). Their structures were identified by extensive spectroscopic analyses, including 1D, 2D NMR and MS and chemical methods. Compound 4 showed cytotoxicity against human hepatoma HepG2 and glioblastoma U87MG cells via inhibition of cell growth and induction of apoptosis. Induction of apoptosis by 4 was demonstrated by cell death, DNA fragmentation, increased Bax/Bcl-2 protein ratio and the activation of caspase-3, caspase-9 and poly (ADP-ribose) polymerase (PARP).

The Suppressive Effects of Integrin Antibodies on the Infection of Hantaan Virus in Fibroblasts (한탄바이러스의 섬유아세포 감염에 대한 Integrin 항체의 억제 효과)

  • Park, Ho-Sun;Kim, Ki-Duk;Kim, Sung-Kwang
    • Journal of Yeungnam Medical Science
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    • v.15 no.1
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    • pp.55-66
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    • 1998
  • Pathophysiological mechanism of hemorrhagic fever with renal syndrome (HFRS) is not fully understood. Major clinical findings of HFRS patients are widespread hemorrhage, acute renal failure and shock. Basic lesion is vascular injury with microvascular hemorrhage and relatively little inflammation. According to autopsy findings, renal medulla shows focal hemorrhage, tubular necrosis and interstitial mononuclear infiltrates. The predominant cell type in the renal and pulmonary interstitium is a fibroblast and it participates in the healing process at the injury site by secreting a large amount of extracellular matrix proteins. Cultured human lung fibroblasts and Mongolian gerbil fibroblasts were known to be good host cells for the hantaan virus. It is possible that not only the endothelial cell but also the fibroblast is a target of Hantaan virus and the fibroblast might be involved in the pathogenesis and the healing process in HFRS. Integrins are adhesion molecules, and act as receptors for many extracellular matrix proteins. Recently, there are many reports that cell surface integrins influence on some viral infections or reversely viruses influence on the expression of integrins. The ${\alpha}_5{\beta}_1$ integrin is a major receptor for the fibronectin which is an important extracellular matrix protein secreted by fibroblasts. In this study, the role of ${\alpha}_5{\beta}_1$ integrin in the infection of Hantaan virus was examined by using anti-${\alpha}_5{\beta}_1$, integrin, anti-${\alpha}_5$ integrin and anti-${\beta}_1$, integrin antibodies in chicken embryo fibroblasts (CEF) and Mongolian gerbil fibroblasts(MGF). The treatment of anti-${\alpha}_5{\beta}_1$, integrin antibody in CEF reduced the virion titers 26.8% and the amount of nucleocapsid N protein 32.6% when compared with control CEF. When MGF were treated with anti-${\alpha}_5$, anti-${\beta}_1$ and anti-${\alpha}_5{\beta}_1$ integrin antibodies, virion titers were reduced by 26.5%, 29.4% and 28.7% and the amount of nucleocapsid N protein were reduced by 65.2%, 59.7% and 72.6%. These results suggested that ${\alpha}_5{\beta}_1$ integrin might act as a receptor for the Hantaan virus or blocking of ${\alpha}_5{\beta}_1$ integrin influences on the viral replication in CEF and MGF. It is also possible that the blocking of only one subunit of integrin represents similar results in that of whole molecule.

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Hydration Properties of $\alpha$-Tricalcium Phosphate in Tris. Solution ($\alpha$-Tricalcium Phosphate의 Tris. Solution에서의 수화특성)

  • 인경필;최상흘
    • Journal of the Korean Ceramic Society
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    • v.30 no.11
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    • pp.905-910
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    • 1993
  • $\alpha$-tricalcium phosphate($\alpha$-TCP) powders were synthesized and their hydration properties were investigated in Tris. solution. Two kinds of $\alpha$-TCP powder samples were prepared; the one is reaction product of CaHPO4.2H2O and CaCO3, and another is that of hydroxyapatite(HAp) and $\beta$-Ca2P2O7. They were satisfied with Ca/P mole ratio 1.5 and were heated at 150$0^{\circ}C$ for 5 hours. In the hydration of $\alpha$-TCP samples the powder which was synthesized from HAp and $\beta$-Ca2P2O7 was hydrated faster than that from CaHPO4.2H2O and CaCO3. The hydration reaction of $\alpha$-TCP powder transformed rapidly into HAp accompanying setting and hardening. It was realized that the hydration reaction of $\alpha$-TCP was due to the solution-precipitation mechanism and the hydrates from the reaction were Ca-deficient HAp having funtional group HPO42-.

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Separation Characteristics of Whey Protein by High Performance Membrane Chromatography (고성능 막 크로마토그래피에 의한 유청 단백질의 분리특성)

  • 홍승범;노경호
    • KSBB Journal
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    • v.16 no.6
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    • pp.533-537
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    • 2001
  • ${\alpha}$-lactalbumin and ${\beta}$-lactoglobulin in whey proteins were separated by high performance membrane chromatography (HPMC). The separation mechanism involved anion-exchange, and the stationary phase was anion CIM (Convective Interaction Media) DEAE, QA disk and cation exchanger SO$_3$(16${\times}$3 mm). Two types of mobile phase were used, buffer A (20 mM Tris-HCI, pH 7.3) and buffer B(buffer A + 1 M NaCl), As the amount of NaCl dissolved in buffer linearly increased, which enabled a gradient elution mode. The optimum mobile phase and operating condition (Buffer A/Buffer B = 100/0 - 30/70 vol%, gradient time 1 min, 30/70 - 10/90 vol.%, gradient time 2 min) were experimentally determined. In this experimental condition, ${\alpha}$-lacta1bumin, ${\beta}$-lactoglobulin were separated within 5 min at a mobile phase flow rate of 4 mL/min.

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Comparison of Volatile Compounds Identified in Different Parts of Peucedanum japonicum Thunberg by Harvest Time (방풍나물의 수확시기에 따른 부위별 향기성분 비교 분석)

  • Jung, Bok-Mi;Shin, Tai-Sun;Heo, Young-Ran
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.12
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    • pp.1871-1880
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    • 2014
  • Headspace volatile compounds of leaf, stem, and root of Peucedanum japonicum Thunberg (PJT) harvested in March and July were isolated, separated, and identified by using a combined system of solid phase microextraction and gas chromatography/mass spectrometry. Identified numbers of volatile compounds were 72, 75, and 63 in March PJT as well as 78, 73, and 69 in July, respectively. Total compounds identified from PJT consisted of alcohols (1~7), aldehydes (7~15), esters (1~4), hydrocarbons (5~9), ketones (1~2), monoterpenes (13~16), monoterpene alcohols (1~5), monoterpene aldehydes (2~4), monoterpene esters (1~4), monoterpene ketones (1~2), sesquiterpenes (16~24), and miscellaneous compounds (2~3). Major volatile compounds of PJT were monoterpenes at all harvest times and in all parts of PJT. The amounts of ${\beta}$-pinene in leaf and stem harvested in March were highest in monoterpenes, followed by ${\alpha}$-pinene and ${\beta}$-myrcene. However, amounts of volatile compounds of monoterpenes in leaf and stem from July PJT were in the decreasing order of ${\beta}$-pinene, ${\beta}$-phellandrene, and ${\beta}$-myrcene. Of the 39 sesquiterpenes identified in PJT, (E)-caryophyllene in March leaf was the most abundant volatile compound, followed by acoradiene in March leaf, ${\beta}$-elemene, and ${\alpha}$-copaene in July stem.

Role of Growth Factors and Cytokines on Bleomycin Induced Pulmonary Fibrosis (Bleomycin 유도 폐 섬유화에 있어서 성장인자 및 Cytokine의 역할)

  • Lee, Yong-Hee;Jung, Soon-Hee;Ahn, Chul-Min;Kim, Sung-Kyu;Cho, Sang-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.871-888
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    • 1997
  • Background : It is now thought that the earliest manifestation of idiopathic pulmonary fibrosis is alveolitis, that is, an accumulation of inflammatory and immune effector cells within alveolar walls and spaces. Inflammatory cells including alveolar macrophages and resident normal pulmonary tissue cells participate through the release of many variable mediators such as inflammatory growth factors and cytokines, which contribute to tissue damage and finally cause chronic pulmonary inflammation and fibrosis. This study was performed to investigate the source and distribution pattern of transforming growth factor-${\beta}_1$(TGF-${\beta}_1$), platelet derived growth factor(PDGF), basic fibroblast growth factor(bFGF), interleukin 1(IL-1), interleukin 6(IL-6), tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and the role of these mediators on bleomycin(BLM)-induced pulmonary injury and fibrosis in rats. Method : Wistar rats were divided into three groups(control group, BLM treated group, BLM and vitamine E treated group). Animals were sacrificed periodically at 1, 2, 3, 4, 5, 7, 14, 21, 28 days after saline or BLM administration. The effects were compared to the results of bronchoalveolar lavage fluid analysis, light microscopic findings, immunohistochemical stains for six different mediators(TGF-${\beta}_1$, PDGF, bFGF, IL-1, IL-6 and TNF-$\alpha$) and mRNA in situ hybridization for TGF-${\beta}_1$. Results : IL-1 and IL-6 are maximally expressed at postbleomycin 1~7th day which are mainly produced by neutrophils and bronchiolar epithelium. It is thought that they induce recruitment of inflammatory cells at the injury site. The expression of IL-1 and IL-6 at the bronchiolar epithelium within 7th day is an indirect evidence of contribution of bronchiolar epithelial cells to promote and maintain the inflammatory and immune responses adjacent to the airways. TNF-$\alpha$ is mainly produced by neutrophils and bronchiolar epithelial cells during 1~5th day, alveolar macrophages during 7~28th day. At the earlier period, TNF-$\alpha$ causes recruitment of inflammatory cells at the injury site and later stimulates pulmonary fibrosis. The main secreting cells of TGF-${\beta}_1$ are alveolar macrophages and bronchiolar epithelium and the target is pulmonary fibroblasts and extracellular matrix. TGF-${\beta}_1$ and PDGF stimulate proliferation of pulmonary fibroblasts and TGF-${\beta}_1$ and bFGF incite the fibroblasts to produce extracellular matrix. The vitamine E and BLM treated group shows few positive cells(p<0.05). Conclusion : After endothelial and epithelial injury, the neutrophils and bronchiolar epithelium secrete IL-1, IL-6, TNF-$\alpha$ which induce infiltration of many neutrophils. It is thought that variable enzymes and $O_2$ radicals released by these neutrophils cause destruction of normal lung architecture and progression of pulmonary fibrosis. At the 7~28th day, TGF-${\beta}_1$, PDGF, bFGF, TNF-$\alpha$ secreted by alveolar macrophages sting pulmonary fibroblasts into proliferating with increased production of extracellular matrix and finally, they make progression of pulmonary fibrosis. TNF-$\alpha$ compares quite important with TGF-${\beta}_1$ to cause pulmonary fibrosis. Vitamine E seems to decrease the extent of BLM induced pulmonary fibrosis.

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Flavor Components of Artemisia Lavandulaefolia DC (참쑥(Artemisia Lavandulaefolia DC)의 방향성분)

  • Choi, Kyoung-Sook;Choi, Bong-Young;Park, Hyoung-Kook;Kim, Jung-Han;Park, Jong-Sei;Yoon, Chang-No
    • Korean Journal of Food Science and Technology
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    • v.20 no.6
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    • pp.774-779
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    • 1988
  • The essential oil of Artemiisa lavandulaefolia DC was analyzed by gas chromatography/mass spectrometry (GC/MS). The oil was stored at different conditions for 6 weeks and the changes of color and chemical composition during storage were checked by GC/MS. The experimental results were as follows; (1) More than 186 chemicals were detected by GC. Major components were 1,8-cineol, thujone, camphor, borneol, coumarin, 2,3-dihydrobenzofuran and ${\beta}-biasbolene$. In this study, 3,3,6-trimethyl norpinanol, ${\beta}-farnesene,\;{\alpha}-curmene$ and 7-methoxy coumarin were detected as new compounds in Artemisia species. (2) It was proved that temperature and/ or light had and important effect on the changes of color and volatile components of the essential oil. The relative amounts of limonene and 1.8-cineol were decreasad, on the other hand, ${\beta}-phellandrene\;and\;{\alpha}-terpineol$ were increased during storage.

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Cytotoxic Constituents of the Leaves of Ixeris sonchifolia

  • Suh, Ji-Young;Jo, Young-Mi;Kim, Nam-Deuk;Bae, Song-Ja;Jung, Jee-H.;Im, Kwang-Sik
    • Archives of Pharmacal Research
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    • v.25 no.3
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    • pp.289-292
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    • 2002
  • The ethyl acetate extract of the leaves of Ixeris sonchifolia afforded two new and two known sesquiterpene lactone glucosides of the guaiane-type, together with a known alkenol glucoside. The known compounds were identified as ixerin Z (1), ixerin Z-6'-p-hydroxyphenylace-tate (2), and (Z)-3-hexen-1-ol-$\beta$-D-glucopyranoside (3), respectively. The structures of the new compounds were elucidated as 11, 13a-dihydroixerin Z [4, 3-hydroxy-2-oxo-guaia-1 (10), $3-dien-5{\alpha},6{\beta},7{\alpha},11{\beta}H-12,6-olide-3-O-{\beta}-D-glucopyranoside],{\;}and{\;}3,10{\$beta}-dihydroxy-2-oxo-guaia-3,11(13)-dien-1{\alpha},5{\alpha},6{\alpha},7aH-12,6-olide-10-O-{\beta}-D-glucopyranoside$ (5), respectively. The cytotoxicity of these compounds against human hepatocellular carcinoma cell (HepG2) and human melanoma cell (SK-MEL-2) was evaluated.

Microstructure Evolution of Ti-6Al-4Fe-0.25Si through Aging Heat Treatment (시효처리에 따른 Ti-6Al-4Fe-0.25Si 합금의 미세조직 변화)

  • Song, Yong Hwan;Kang, Joo-Hee;Park, Chan Hee;Kim, Seong-Woong;Hyun, Yong-Taek;Kang, Nam Hyun;Yeom, Jong-Taek
    • Korean Journal of Metals and Materials
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    • v.50 no.7
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    • pp.477-485
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    • 2012
  • The effect of aging heat treatment on microstructure evolution of the Ti-6Al-4Fe-0.25Si alloy with an initial microstructure of an elongated alpha was investigated. Aging treatments of the samples were carried out at $550^{\circ}C$ for up to 100 hours. The microstructure of the 5 hours heat-treated sample consisted of alpha grains, beta matrix and some TiFe intermetallic compounds that were precipitated from the beta matrix. Increasing the aging time to 10 hours, most of the beta matrix was decomposed to very fine alpha grains (${\sim}0.5{\mu}m$) and TiFe, and thus the volume fraction of the beta matrix was significantly decreased. EBSD analysis revealed that newly formed tertiary-alpha-grains in the vicinity of TiFe had high angle boundaries with respect to the primary and secondary alpha grains. As a result of these phase transformations during aging, the fraction of the alpha/alpha grain boundary was increased while that of the alpha/beta phase boundary was decreased.