• Title/Summary/Keyword: $6{\beta}$

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Characterization of the \beta-Galactosidase Produced by Streptomyces sp. YB-10 (\beta-Galactosidase를 생산하는 Streptomyces sp. YB-10의 분리 및 효소 특성)

  • 윤기홍;이경섭;김창진
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.151-156
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    • 2003
  • A strain YB-10 was isolated from soil as a producer of the extracellular $\beta$-D-galactosidase, which catalyzes the hydrolysis of lactose. The strain YB-10 was identified as Streptomyces sp. on the basis of its cultural, morphological and physiological properties. After treating culture supernatant of the isolate with ammonium sulfate, the precipitated protein was used as a crude $\beta$-galactosidase for analyzing its reaction properties with para-nitrophenyl-$\beta$-D-galactosidase(pNP-$\beta$Gal) as a substrate. The $\beta$-galactosidase showed its maximal activity at pH 6.0 and 6$0^{\circ}C$. The enzyme was also active on lactose. The hydrolyzing activity of $\beta$-galactosldase for pNP-$\beta$Gal and lactose was decreased by galactose. Its hydrolyzing activity far lactose was also decreased by glucose, but the activity for pNP-$\beta$Gal was increased to 1.8-folds by glucose.

The Effects of Proinflammatory Cytokines and TGF-beta, on The Fibroblast Proliferation (Proinflammatory Cytokines과 TGF-beta가 섬유모세포의 증식에 미치는 영향)

  • Kim, Chul;Park, Choon-Sik;Kim, Mi-Ho;Chang, Hun-Soo;Chung, Il-Yup;Ki, Shin-Young;Uh, Soo-Taek;Moon, Seung-Hyuk;Kim, Yong-Hoon;Lee, Hi-Bal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.861-869
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    • 1998
  • Backgrounds: The injury of a tissue results in the infalmmation, and the imflammed tissue is replaced by the normal parenchymal cells during the process of repair. But, constitutional or repetitive damage of a tissue causes the deposition of collagen resulting in the loss of its function. These lesions are found in the lung of patients with idiopathic pulmonary fibrosis, complicated fibrosis after diffuse alveolar damage (DAD) and inorganic dust-induced lung fibrosis. The tissue from lungs of patients undergoing episodes of active and/or end-stage pulmonary fibrosis shows the accumulation of inflammatory cells, such as mononuclear cells, neutrophils, mast cells and eosinophils, and fibroblast hyperplasia. In this regard, it appears that the inflammation triggers fibroblast activation and proliferation with enhanced matrix synthesis, stimulated by inflammatory mediators such as interleukin-1 (IL-1) and/or tumor necrosis factor (TNF). It has been well known that TGF-$\beta$ enhance the proliferation of fibroblasts and the production of collagen and fibronectin, and inhibit the degradation of collagen. In this regard, It is likely that TGF-$\beta$ undergoes important roles in the pathogenesis of pulmonary fibrosis. Nevertheless, this single cytokine is not the sole regulator of the pulmonary fibrotic response. It is likely that the balance of many cytokines including TGF-$\beta$, IL-1, IL-6 and TNF-$\alpha$ regulates the pathogenesis of pulmonary fibrosis. In this study, we investigate the interaction of TGF-$\beta$, IL-1$\beta$, IL-6 and TNF-$\alpha$ and their effect on the proliferation of fibroblasts. Methods: We used a human fibroblast cell line, MRC-5 (ATCC). The culture of MRC-5 was confirmed by immunofluorecent staining. First, we determined the concentration of serum in cuture medium, in which the proliferation of MRC-5 is supressed but the survival of MRC-5 is retained. Second, we measured optical density after staining the cytokine-stimulated cells with 0.5% naphthol blue black in order to detect the effect of cytokines on the proliferation of MRC-5. Result: In the medium containing 0.5% fetal calf serum, the proliferation of MRC-5 increased by 50%, and it was maintained for 6 days. IL-1$\beta$, TNF-$\alpha$ and IL-6 induced the proliferation of MRC-5 by 45%, 160% and 120%, respectively. IL-1$\beta$ and TNF-$\alpha$ enhanced TGF-$\beta$-induced proliferation of MRC-5 by 64% and 159%, but IL-6 did not affect the TGF-$\beta$-induced proliferation. And lNF-$\alpha$-induced proliferation of MRC-5 was reduced by IL-1$\beta$ in 50%. TGF-$\beta$, TNF-$\alpha$ and both induced the proliferation of MRC-5 to 89%, 135% and 222%, respectively. Conclusions: TNF-$\alpha$, TGF-$\beta$ and IL-1$\beta$, in the order of the effectiveness, showed the induction of MRC-5 proliferation of MRC-5. TNF-$\alpha$ and IL-1$\beta$ enhance the TGF-$\beta$-induced proliferation of MRC-5, but IL-6 did not have any effect TNF-$\alpha$-induced proliferation of MRC-5 is diminished by IL-1, and TNF-$\alpha$ and TGF-$\beta$ showed a additive effect.

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A Study on the Constituents from the Roots of Polygala tenuifolia (원지(Polygala tenuifolia WILLD.) 뿌리의 성분연구)

  • Lee, Young-Sun;Lee, Je-Hyun;Kim, Chung-Sook;Kim, Jin-Sook
    • Korean Journal of Pharmacognosy
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    • v.30 no.2
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    • pp.168-172
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    • 1999
  • Five compounds were isolated from the roots of Polygala tenuifolia (Polygalaceae). On the basis of spectroscopic evidences, the structures of these compounds were characterized as ${\alpha}-D-(6-O-sinapoyl)-glucopyranosyl(1{\rightarrow}2')-{\beta}-D-(3'-O-sinapoyl)-fructofuranoside$ (P3), ${\alpha}$-D-{6-O-(p-methoxybenzoyl)}-glucopyranosyl-$(1{\rightarrow}2')$-${\beta}$-D-{3'-O-(3',4',5'-trimethoxycinnamoyl)}-fructofuranoside(P4), ${\alpha}$-D-{6-O-(p-hydroxybenzoyl)}-glucopyranosyl-$(1{\rightarrow}2')$-${\beta}$-D-{3'-O-(3',4',5'-trimethoxycinnamoyl)}-fructofuranoside(P5), ${\alpha}-D-glucopyranosyl-(1{\rightarrow}2')-{\beta}-D-(1'-O-sinapoyl)-fructofuranoside$(P6), $1,5-anhydro-D-glucitol$(P7) respectively. ${\alpha}$-D-{6-O-(p-Methoxybenzoyl)}-glucopyranosyl-$(1{\rightarrow}2')$-${\beta}$-D-{3'-O-(3',4',5'-trimethoxycinnamoyl)}-fructofuranoside(P4) and ${\alpha}-D-glucopyranosyl-(1{\rightarrow}2')-{\beta}-D-(1'-O-sinapoyl)-fructofuranoside$(P6) were isolated for the first time from the genus of Polygala. 1,5-Anhydro-D-glucitol(P7) was isolated without hydrolysis for the first time from the root of Polygala tenuifolia.

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Conformation of cyclo-[Gln-Trp-Phe- $\beta$Ala-Leu-Met], a NK-2 Tachykinin Receptor Antagonist (NK-2의 Antagonist인 cyclo-[Gln-Trp-Phe- $\beta$Ala-Leu-Met]의 형태에 관한 연구)

  • Ha, Jong Myung
    • Journal of the Korean Chemical Society
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    • v.43 no.5
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    • pp.540-546
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    • 1999
  • Solution conformation of cyclo-($Gln^1-Trp^2-Phe^3-{\beta}Ala^4-Leu^5-Met^6$), new NK-2 antagonist in dimethyl sulfoxide solution, has been determined by the use of two-dimensional nuclear magnetic resonance spectroscopy combined with simulated annealing calculations. The peptide exhibited converged structures with the atomic root-mean-square difference for the backbone atoms ($N,\;C^{\alpha},\;C'$) of all residues being 0.02${\AA}$ in the 25 annealed structures. The analysis of the structures indicated that the cyclic peptide has three intramolecular hydrogen bonds between $Met^6NH$ and ${\beta}Ala^4CO$, ${\beta}Ala^4NH$ and $Met^6CO$, $Phe^3NH$ and $Met^6CO$, and contain a type-I ${\beta}$-turn with Gln and Trp and ${\gamma}$-turn with Leu. The addition of an extra methylene group to Gly, i.e. P-Ala residue, may relax some unfavorable restraints in the cyclic backbone structure, hence enabling an additional hydrogen bond, which results in stabilizing one conformation.

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Physicochemical Characteristics of Pound Cake Added with $\beta-Glucan$ ($\beta-Glucan$ 첨가 Pound Cake의 이화학적$\cdot$관능적 특성)

  • Shin Yu-Mi;Kim Mi-Kyoung;Cho Han-Young;Kim Mee-Ree
    • Journal of the East Asian Society of Dietary Life
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    • v.15 no.6
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    • pp.728-737
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    • 2005
  • This study was conducted to evaluate the physicochemical characteristics of pound cake made of substituted flour with different amount of $\beta-glucan$ i.e. 3, 6 and $9\%$ respectively. $\beta-glucan$ is a functional food material produced from Agrobacterium sup. R259 KCTC 10197BP. The specific gravity and pH of dough were found to be increased according to $\beta-glucan$ content Rheological property such as adhesiveness of dough has no difference between control and $\beta-glucan$ added groups. The pH of baked pound cake was not significantly different between control and $\beta-glucan$ groups. The volume of baked pound cake wag not significantly different between control and the ones with up to $6\%$ $\beta-glucan$, Lightness (L), redness (a) and yellowness (b) value of Hunter color system of cake crumb were not significantly different among teated groups, except a value of crust in $9\%$ $\beta-glucan$ group. the moisture content of cake increased according to $\beta-glucan$ content. Hardness by texture analyser was decreased with increasing $\beta-glucan$ content although cohesiveness wag not changed in β$\beta-glucan$ groups, compared with the control. Sensory evaluation results showed that the mean scores of over·all acceptability in 3 and $6\%$ $\beta-glucan$ were higher than that of control. Based on these results, the addition of $\beta-glucan$ to pound cake up to $6\%$ was considered to be desirable especially in terms of texture.

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Characterization of the Extracellular ${\beta}-Galactosidase$ Produced from Streptomyces sp. YB-9 (Streptomyces sp. YB-9가 생산하는 균체외 ${\beta}-galactosidase$의 특성)

  • Lee, Kyung-Seop;Kim, Chang-Jin;Yoon, Ki-Hong
    • Applied Biological Chemistry
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    • v.46 no.4
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    • pp.299-304
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    • 2003
  • A strain YB-9 was isolated from soil as a producer of the extracellular ${\beta}-D-galactosidase$, which catalyzes the hydrolysis of lactose. The strain YB-9 was identified as Streptomyces sp. on the basis of its cultural, morphological and physiological properties. After treating culture supematant of the isolate with ammonium sulfate $(15{\sim}70%)$, the precipitated protein was used as a crude ${\beta}-galactosidase$ for analyzing its reaction properties with $para-nitrophenyl-{\beta}-D-galactoside$ $(pNP-{\beta}Gal)$ and lactose as substrates. The {\beta}-galactosidase showed its maximal activity at pH $6.0{\sim}6.5$ and $60^{\circ}C$. The hydrolyzing activity of ${\beta}-galactosidase$ for both $pNP-{\beta}Gal$ and lactose was decreased by galactose. Its hydrolyzing activity for lactose was slightly decreased by glucose, but the activity for $pNP-{\beta}Gal$ was increased to 1.3-folds by glucose. Especially, its hydrolyzing activity was not affected for lactose and was increased to 1.6-folds for $pNP-{\beta}Gal$ by xylose.

Immunomodulatory Effects of β-1,3/1,6-glucan and Lactobacillus plantarum LM1004 on Atopic Dermatitis Models (아토피 피부염 모델에 대한 β-1,3/1,6-glucan과 Lactobacillus plantarum LM1004의 면역 조절 효과)

  • Kim, In Sung;Kim, Sung Hak;Kim, Jeong A;Yu, Da Yoon;Kim, Gwang Il;Park, Dong-Chan;Lim, Jong Min;Lee, Sang Suk;Choi, In Soon;Cho, Kwang Keun
    • Journal of Life Science
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    • v.28 no.1
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    • pp.17-25
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    • 2018
  • In this study, we examined the efficacy of the immune regulation of ${\beta}$-1,3/1,6-glucan and Lactobacillus plantarum LM1004 on atopic dermatitis models. The oral administration of ${\beta}$-1,3/1,6-glucan and L. plantarum LM1004 on mice significantly decreased the amount of scratching, leakage to evans blue, and concentrations of serum immunoglobulin E (IgE) and histamine compared with the atopic dermatitis - induced group. When atopic dermatitis was induced, the transcription factors (GATA-3, retinoic acid-related orphan receptor ${\gamma}$ T [$ROR{\gamma}T$]) and cytokines (interleukin-4 [IL-4], IL-17) of Th2 and Th17 cells were overexpressed at the transcriptional level, and they significantly decreased with oral administration of ${\beta}$-1,3/1,6-glucan and L. plantarum LM1004. In addition, ${\beta}$-1,3/1,6-glucan and L. plantarum LM1004 were shown to modulate the immune balance by increasing the expression of Th1 and Treg transcription (T-bet, forkhead box p3 [Foxp3]) and cytokines (interferon-${\gamma}$ [IFN-${\gamma}$], transforming growth factor-${\beta}$ [TGF-${\beta}$]). Galectin-9 and filaggrin were significantly lower in the atopic dermatitis - induced group and significantly higher in the ${\beta}$-1,3/1,6-glucan-treated group. In contrast, thymic stromal lymphopoietin (TSLP) was highest in the atopic dermatitis-induced group, while mice that were orally administered ${\beta}$-1,3/1,6-glucan and L. plantarum LM1004 showed similar TSLP levels to the control group. These results indicate that ${\beta}$-1,3/1,6-glucan and L. plantarum LM1004 have immunomodulatory effects and atopic dermatitis improvement effects in an animal model of atopic dermatitis. Therefore, it is expected that ${\beta}$-1,3/1,6-glucan and L. plantarum LM1004 can be used as natural materials in the treatment of atopic dermatitis.

Tubulin Beta3 Serves as a Target of HDAC3 and Mediates Resistance to Microtubule-Targeting Drugs

  • Kim, Youngmi;Kim, Hyuna;Jeoung, Dooil
    • Molecules and Cells
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    • v.38 no.8
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    • pp.705-714
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    • 2015
  • We investigated the role of HDAC3 in anti-cancer drug-resistance. The expression of HDAC3 was decreased in cancer cell lines resistant to anti-cancer drugs such as celastrol and taxol. HDAC3 conferred sensitivity to these anti-cancer drugs. HDAC3 activity was necessary for conferring sensitivity to these anti-cancer drugs. The down-regulation of HDAC3 increased the expression of MDR1 and conferred resistance to anti-cancer drugs. The expression of tubulin ${\beta}3$ was increased in drug-resistant cancer cell lines. ChIP assays showed the binding of HDAC3 to the promoter sequences of tubulin ${\beta}3$ and HDAC6. HDAC6 showed an interaction with tubulin ${\beta}3$. HDAC3 had a negative regulatory role in the expression of tubulin ${\beta}3$ and HDAC6. The down-regulation of HDAC6 decreased the expression of MDR1 and tubulin ${\beta}3$, but did not affect HDAC3 expression. The down-regulation of HDAC6 conferred sensitivity to taxol. The down-regulation of tubulin ${\beta}3$ did not affect the expression of HDAC6 or MDR1. The down-regulation of tubulin ${\beta}3$ conferred sensitivity to anti-cancer drugs. Our results showed that tubulin ${\beta}3$ serves as a downstream target of HDAC3 and mediates resistance to microtubule-targeting drugs. Thus, the HDAC3-HDAC6-Tubulin ${\beta}$ axis can be employed for the development of anti-cancer drugs.

The Production and Correlation of Silica Induced Proinflammatory Cytokines and TGF-$\beta$ from Monocytes of Balb/C Mice (Balb/C mouse의 폐장대식세포에서 유리규산 자극에 의한 Proinflammatory Cytokine과 TGF-$\beta$의 생성 및 상관관계)

  • Ki, Shin-Young;Kim, Eun-Young;Kim, Mi-Ho;Uh, Soo-Taek;Kim, Yong-Hoon;Park, Choon-Sik;Lee, Hi-Bal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.823-834
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    • 1998
  • Background: Chronic inhalation of silica induces the lung fiborsis. The alveolar macrophages ingest the inhaled silica; they liberate the pro-inflammatory cytokines such as IL-1$\beta$, IL-6, TNF-$\alpha$ and fibrogenic cytokines, TGF-$\beta$ and PDGF. Cytokines liberated from macrophage have pivotal role in pulmonary fibrosis. There is a complex cytokine network toward fibrosis. However, the exact roles and the interaction among the proinflammatory cytokines and TGF-$\beta$, a fibrogenic cytokine, have not been defined, yet. In this study, we investigated silica induced IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ production and the effect of IL-1$\beta$, IL-6, TNF-$\alpha$ on the production of TGF-$\beta$ from lung macrophages of Balb/C mice. Method: We extracted the lung of Balb/C mice and purified monocytes by Percoll gradient method. Macrphages were stimulated by silica ($SiO_2$) in the various concentration for 2, 4, 8, 12, and 24 hours. The supernatants were used for the measurement of protein levels by bioassay, and cells for the levels of mRNA by in situ hybridization. Results: The production of IL-6 was not observed till 4 hours, and reached the peak levels at 8 hours after stimulation of silica. The production of TNF-$\alpha$ increased from 2 hours and reached the peak levels at 4 hours after stimulation of silica. The spontaneous TGF-$\beta$ production reached the peak levels at 24 hours. TNF-$\alpha$ upregulated the silica induced TGF-$\beta$ production. Silica induced TGF-$\beta$ production was blocked by pretreated anti-TNF-$\alpha$ antibody. In situ hybridization revealed the increased positive signals at 4 hours in IL-6, at 4 hours TNF-$\alpha$ and 12 hours in TGF-$\beta$. Conclusion: The results above suggest that silica induced the sequential production of IL-6, 1NF-$\alpha$ and TGF-$\beta$ from macrophages and TNF-$\alpha$ upregultaes the production of TGF-$\beta$ from silica-induced macrophages.

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Direct Syntheses of $\beta-Mannopyranosyl$ Disaccharides from 4,6-O-Benzylidene Derivatives of Ethylthio $\alpha-D-Mannopyranosides$ Donors

  • Yun, Mi Gyeong;Sin, Yeong Suk;Cheon, Geun Ho
    • Bulletin of the Korean Chemical Society
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    • v.21 no.6
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    • pp.562-566
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    • 2000
  • $\beta-D-Mannopyranosyl$ disaccharides have been obtained from the coupling of 4,6-O-benzylidene derivatives of ethylthio $\alpha-D-mannopyranoside$ employing NIS-TfOH promoter. NIS-TfOH promoted couplings of the corresponding ethylthio $\beta-D-glucopyranoside$ and produced $\alpha-D-glucopyranosyl$ disaccharides. IDCP (iodonium dicollidine perchlorate) was inactive toward the 4,6-O-benzylidenated ethylthio glucopyranosyl donor. However,lDCP coupled the 4,6-O-benzylidenated $ethylthio-\beta-D-galactopyranoside$ to give a-D-galactopyranosyl disaccharides.