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Bhang, Suk Ho,Jeon, Jeong-Yi,La, Wan-Geun,Seong, Jun Yeup,Hwang, Jin Wook,Ryu, Seong Eon,Kim, Byung-Soo Published for the International Union of Biochemis 2011 Biotechnology and Applied Biochemistry Vol.58 No.4
<P>This study was designed to evaluate the additive effects of transforming growth factor-beta3 (TGF-β3) and hyaluronic acid (HA) on chondrogenic differentiation of human mesenchymal stem cells (hMSCs). The hMSCs were cultured on collagen type I-, HA-, or fibronectin-coated cell culture dishes with or without TGF-β3 added to the culture medium. Four weeks after cell culture, chondrogenic differentiation of hMSCs was determined by evaluating the expression of cartilage-specific markers using real-time polymerase chain reaction, immunocytochemistry, and Western blot analysis. hMSCs cultured on HA-coated dishes with TGF-β3 supplementation revealed a prominent increase in collagen type II, aggrecan, and Sox9. When hMSCs were cultured without TGF-β3 supplementation, only hMSCs cultured on HA-coated dishes showed prominent expression of the cartilage-specific markers. This study shows that chondrogenic differentiation of hMSCs can be enhanced additively by interactions with both a specific cell-adhesion matrix and a soluble growth factor.</P>
Increased 2,3-butanediol production by changing codon usages in Escherichia coli.
Park, Seo-Young,Kim, Borim,Lee, Soojin,Oh, Minkyu,Won, Jong-In,Lee, Jinwon Published for the International Union of Biochemis 2014 Biotechnology and Applied Biochemistry Vol.61 No.5
<P>The natural microorganism Escherichia coli without modification is not suitable for the efficient production of 2,3-butanediol (2,3-BD) on an industrial scale because of its poor metabolic performance. Metabolic capacities of E. coli have been improved to produce 2,3-BD efficiently, the performance of which is possible for producing such a product. Codon optimization with the ribosome-binding site for the efficient production of target genes (budA and budC) was achieved by molecular engineering, which allowed the metabolic engineering to proceed to the next level. As a result, comparing the productivity in 26?H, where the amount of p18COR was 1.04?g/L and that of p18WTR was 0.41?g/L, represents an approximate 60.6% increase in the productivity of the p18WTR with codon optimization. In other words, p18COR was 2.54-fold greater than p18WTR in the production of 2,3-BD.</P>
An antifungal mechanism of curcumin lies in membrane-targeted action within Candida albicans.
Lee, Wonyoung,Lee, Dong Gun Published for the International Union of Biochemis 2014 IUBMB life Vol.66 No.11
<P>The aim of this study is to investigate the antifungal mechanism of curcumin. This polyphenolic compound has been used traditionally in Asia for medicinal, culinary, and other purposes. Although antifungal effect of curcumin has been reported, this is the first study for its mode of action underlying disruption of plasma membrane in Candida albicans. The leakage of potassium ion from the fungal cytosol and dissipation in membrane potential was detected by bis-(1,3-dibutylbarbituric acid)trimethine oxonol [DiBAC4 ] staining. We also investigated an increase in membrane permeability in curcumin-treated C. albicans with influx of propidium iodide assay. Fluorescence analysis with 1,6-diphenyl-1,3,5-hexatriene supported the membrane-targeted mechanism of action indicating membrane disruption. On the basis of these results, we studied the effects of curcumin treatment on model membrane to elucidate its antifungal mechanism. Using calcein leakage assays from curcumin-treated large unilamellar vesicles and giant unilamellar vesicles, we found that curcumin has membrane-active mechanism inducing leakage of intracellular component through the flappy membrane. Therefore, this study suggests that curcumin exerts antifungal activity via inducing disruption of fungal plasma membrane.</P>
Biosynthesis of drosopterins, the red eye pigments of Drosophila melanogaster.
Kim, Heuijong,Kim, Kiyoung,Yim, Jeongbin Published for the International Union of Biochemis 2013 IUBMB life Vol.65 No.4
<P>Drosophila melanogaster has red eyes. Scientists have been curious about the biosynthesis of the red eye pigments and have completed a number of investigations on these compounds. Scientific contributions made over the past 50 years have improved our understanding of the red eye pigments. Researchers have elucidated the chemical structures of some pigments and have successfully purified and identified the enzymes that participate in the biosynthesis of the red eye pigments. In this article, we will review the characteristics of the Drosophila red eye pigments and of the enzymes and genes involved in its biosynthetic pathway.</P>
Park, Yoorim,Jung, Min Kyung,Yoon, Sun Young,Lee, Ha-Reum,Hur, Dae Young,Kim, Daejin,Yang, Yoolhee,Kim, Tae Sung,Kim, Seonghan,Yoon, Suk Ran,Park, Hyun Jeong,Bang, Sa Ik,Cho, Dae Ho Published for the International Union of Biochemis 2013 Biotechnology and applied biochemistry Vol.60 No.3
<P>Adipose stem cells (ASCs) are pluripotent cells that can generate pure fat tissue for regeneration. Differentiated adipose cells have been generated by a common inducer cocktail composed of dexamethasone, insulin, and isobutylmethylxanthine (DIM). The major drawbacks of adipose cells are their tendency to float on the culture media and their cost. To overcome some of these disadvantages, a new inducer cocktail that includes insulin, dehydroepiandrosterone, and histamine (DH IH) was tested. As a result, lipid accumulation was elevated more than twofold with DH IH than with DIM. Cell adhesion and viability, which are important factors for stable differentiation, were increased with DH IH and were proven through measurement of mRNA expression levels of adhesion marker genes, N-cadherin and vascular cell adhesion molecule, as well as through an alamar blue assay. The expression of adipogenesis-related genes, adiponectin, and glucose transporter type 4 lasted for a long time. To improve the efficiency of grafting, cell adhesion and neovascularization need to be increased. Neovascularization was observed around the transplanted adipose cells, which showed a higher number of vessel formation in DH IH than in DIM. The above results suggest that DH IH can produce pure differentiated adipose cells effectively and enhance their adhesion onto the target location when these differentiated adipose cells were applied as a clinical resource.</P>
Park, Sang Gyu,Choi, Eung-Chil,Kim, Sunghoon Published for the International Union of Biochemis 2010 IUBMB life Vol.62 No.4
<P>Aminoacyl-tRNA synthetases (ARSs) are highly conserved for efficient and precise translation of genetic codes. In higher eukaryotic systems, several different ARSs including glutamyl-prolyl-, isoelucyl-, leucyl-, methionyl-, glutaminyl-, lysyl-, arginyl-, and aspartyl-tRNA synthetase form a macromolecular protein complex with three nonenzymatic cofactors (AIMP1/p43, AIMP2/p38, and AIMP3/p18). Although the structure and functional implications for this complex formation are not completely understood, rapidly accumulating evidences suggest that this complex would work as a molecular hub linked to the multiple signaling pathways that involve the components of enzymes and cofactors. In this article, the roles of three nonenzymatic components of the multi-tRNA synthetase complex in the assembly of the components and in cell regulation are addressed.</P>
Lee, Chang Hoon,Woo, Jung Hee,Cho, Kwang Keun,Kang, Seung Ha,Kang, Sang Kee,Choi, Yun Jaie Published for the International Union of Biochemis 2007 Biotechnology and Applied Biochemistry Vol.46 No.4
<P>The major obstacle for oral delivery of administered therapeutic proteins is malabsorption in the intestine. This malabsorption could be overcome by induction of neonatal FcRn [Fc (CH2 and CH3 domains of human IgG1 antibody) receptor]-mediated transcytosis in the intestine using recombinant fusion of CH2 and CH3 moieties of human IgG to a therapeutic protein. To this end we developed recombinant hGH (human growth hormone) fused to the N-terminus of Fc moieties [CH2-CH3 or h (hinge)-CH2-CH3] from human IgG1. These recombinant proteins secreted by the methylotrophic yeast Pichia pastoris functionally induced secretion of insulin-like growth factor 1 by HepG2 cells in the response to hGH moiety in the fusion proteins. In a transport study using polarized T84 cells, 3.7% of added dimeric hGH-h-Fc was transported in the apical-to-basolateral direction within 1 h by FcRn-mediated transcytosis of 1 cm(2) monolayers. However, transport of monomeric hGH-Fc (only 0.43%) was much less effective, yet its transport was 2.3 times higher than that of hGH. Finally, we concluded that, upon recombinant fusion, maintenance of dimeric structure of Fc moieties is crucial for the induction of FcRn-mediated transcytosis.</P>