• Title/Summary/Keyword: stable cells

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Production of NADP by Immobilized Brevibacterium ammoniagenes and ATP- regenerating System of Acetate Kinase (고정화 Brevibacterium ammoniagenes와 Acetate Kinase의 ATP생성계에 의한 NADP생산)

  • 조정일
    • The Korean Journal of Food And Nutrition
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    • v.6 no.3
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    • pp.158-168
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    • 1993
  • For the conversion of WAD to NADP, Immobilized Brevibacterium ammoniagenes cells with NAD kinase was coupled with ATP-generating system by acetate kinase. The membrane permeability of B. ammoniagenes was improved by toluene treatment of cells. The toluene treated B. ammoniagenes cells were immobilized for stable enzyme activity. Partially purified acetate kinase was used in the reaction system. The optimum conditions for the efficient conversion of UAD to WADP by energy-coupled system were investigated. B. ammoniagenes cells treated with toluene for the Improvement of membrane permeability showed 4.5 fold improved permeability in the conversion of NAD to NADP compared with Intact cells. 3% k-carrageenan as the immobilization matrix of B. ammoniagenes showed the best efficiency for the conversion of NAD to NADP The optimum conditions for the WAR to WARP conversion reaction coupled nth ATP-generating system were 10mM acetylphosphate, 5mM ADP 200mM inorganic phosphate, 10mM MgCl2, 250mg/ml Immobilized cells, 49.3mUnit/ml acetate kinase, pH 7.5 and 37$^{\circ}C$. Under the optimum conditions, 72% of 5mM(340mg/ml ) NAD was converted to UADP In 12 hours.

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Apoptotic Cell Death in TrkA-overexpressing Cells: Kinetic Regulation of ERK Phosphorylation and Caspase-7 Activation

  • Jung, Eun Joo;Kim, Deok Ryong
    • Molecules and Cells
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    • v.26 no.1
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    • pp.12-17
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    • 2008
  • The TrkA tyrosine kinase is activated by autophosphorylation in response to NGF, and plays an important role in cell survival, differentiation, and apoptosis. To investigate its role in cell fate determination, we produced stable TrkA-inducible SK-N-MC and U2OS cell lines using the Tet-On system. Interestingly, TrkA overexpression induced substantial cell death even in the absence of NGF, by stimulating ERK phosphorylation and caspase-7 activation leading to PARP cleavage. TrkA-mediated cell death was shown by the annexin-V binding assay to be, at least in part, apoptotic in both SK-N-MC and U2OS cells. Furthermore, the truncated form (p18) of Bax accumulated in the TrkA-induced cells, suggesting that TrkA induces mitochondria-mediated apoptosis. NGF treatment augmented the cell death induced by TrkA overexpression. This TrkA-induced cell death was blocked by the tyrosine kinase inhibitors, K-252a and GW441756. Moreover, TrkA overexpression inhibited long-term proliferation of both the neuronal SK-N-MC cells and the non-neuronal U2OS cells, suggesting a potential role of TrkA as a tumor suppressor.

Experimental Investigation on High Efficient Electrolytes of Electrochemical Photovoltaic Cells (전기화학형 광전변환 셀의 고효율 전해질 제작에 관한 실험적 고찰)

  • Kim, Doo-Hwan;Han, Chi-Hwan;Sung, Youl-Moon
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.60 no.1
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    • pp.100-104
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    • 2011
  • In this work, an optimum condition of electrolytes preparation for photovoltaic cells application was investigated experimentally in terms of impedance and conversion efficiency of the cells. 3-methoxyppropionitrie and redox pairs with LiI and $I_2$ were used as stable solvents for fabrication of electrolyte. Efficiency comparison of the prepared cells carried out for various additives and ionic liquids. From the results, there was an optimum concentration (about 0.3 M) of ionic liquids for efficient cell fabrication. For case of electrolyte using single DMAp additive, the maximum conversion efficiency of the cell was 6.4%($V_{oc}$: 0.78V, $J_{sc}$: 14.4 mA/$cm^2$, ff: 0.57). For case of electrolyte using both DMAp and CEMim additives, the maximum conversion efficiency of the cell was 7.2%($V_{oc}$: 0.79V, $J_{sc}$: 16 mA/$cm^2$, ff: 0.57). From the result of electrochemical impedance measurement, both Z1 and Z3 values of binary additives-based cell decreased compared to those of single additive-based. This is due to the decreased in internal and charge transfer resistivities of the cells.

Effects of programmed cell death induction method on somatic cell development

  • Kim, Sang-Hwan
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.3
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    • pp.137-144
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    • 2021
  • In this study, to analyze whether artificial regulation of apoptosis in the development of somatic cells can affect the stable growth and development of cells, 20 alpha-hydroxysteroid dehydrogenase (20α-HSD) and rapamycin were treated to induce apoptosis and autophagy in the both skin and muscle cells. Respectively, and 3-methyladenine was supplemented to inhibit cell death. Our results show that stimulation with rapamycin activated autophagy in both types of cells, but increased apoptosis more than autophagy in the case of skin cells. These results indicate that there was a difference in the expression of survival factors and cell development depending on the type of cell. In particular, in the expression of autophagy-related gene (MAP1LC3A) was higher than that of Casp-3, an apoptosis factor. Furthermore, cell development was the highest in all cell groups cultured by artificially inducing autophagy, however the lowest in the apoptosis-inhibited group. Especially, the noteworthy result of this study was that when apoptosis was induced using 20α-HSD, it was possible to induce apoptosis in both skin and muscle cells. Therefore, the main point of this study is that apoptosis induced during cell culture plays a pivotal role in cell remodeling.

TATA box binding protein and ribosomal protein 4 are suitable reference genes for normalization during quantitative polymerase chain reaction study in bovine mesenchymal stem cells

  • Jang, Si-Jung;Jeon, Ryoung-Hoon;Kim, Hwan-Deuk;Hwang, Jong-Chan;Lee, Hyeon-Jeong;Bae, Seul-Gi;Lee, Sung-Lim;Rho, Gyu-Jin;Kim, Seung-Joon;Lee, Won-Jae
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.12
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    • pp.2021-2030
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    • 2020
  • Objective: Quantitative polymerase chain reaction (qPCR) has been extensively used in the field of mesenchymal stem cell (MSC) research to elucidate their characteristics and clinical potential by normalization of target genes against reference genes (RGs), which are believed to be stably expressed irrespective of various experimental conditions. However, the expression of RGs is also variable depending on the experimental conditions, which may lead to false or contradictory conclusions upon normalization. Due to the current lack of information for a clear list of stable RGs in bovine MSCs, we conducted this study to identify suitable RGs in bovine MSCs. Methods: The cycle threshold values of ten traditionally used RGs (18S ribosomal RNA [18S], beta-2-microglobulin [B2M], H2A histone family, member Z [H2A], peptidylprolyl isomerase A [PPIA], ribosomal protein 4 [RPL4], succinate dehydrogenase complex, subunit A [SDHA], beta actin [ACTB], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], TATA box binding protein [TBP], and hypoxanthine phosphoribosyltrasnfrase1 [HPRT1]) in bovine bone marrow-derived MSCs (bBMMSCs) were validated for their stabilities using three types of RG evaluation algorithms (geNorm, Normfinder, and Bestkeeper). The effect of validated RGs was then verified by normalization of lineage-specific genes (fatty acid binding protein 4 [FABP4] and osteonectin [ON]) expressions during differentiations of bBMMSCs or POU class 5 homeobox 1 (OCT4) expression between bBMMSCs and dermal skins. Results: Based on the results obtained for the three most stable RGs from geNorm (TBP, RPL4, and H2A), Normfinder (TBP, RPL4, and SDHA), and Bestkeeper (TBP, RPL4, and SDHA), it was comprehensively determined that TBP and RPL4 were the most stable RGs in bBMMSCs. However, traditional RGs were suggested to be the least stable (18S) or moderately stable (GAPDH and ACTB) in bBMMSCs. Normalization of FABP4 or ON against TBP, RPL4, and 18S presented significant differences during differentiation of bBMMSCs. However, although significantly low expression of OCT4 was detected in dermal skins compared to that in bBMMSCs when TBP and RPL4 were used in normalization, normalization against 18S exhibited no significance. Conclusion: This study proposes that TBP and RPL4 were suitable as stable RGs for qPCR study in bovine MSCs.

Analysis of the Molecular Event of ICAM-1 Interaction with LFA-1 During Leukocyte Adhesion Using a Reconstituted Mammalian Cell Expression Model

  • Han, Weon-Cheol;Kim, Kwon-Seop;Park, Jae-Seung;Hwang, Sung-Yeoun;Moon, Hyung-Bae;Chung, Hun-Taeg;Jun, Chang-Duk
    • Animal cells and systems
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    • v.5 no.3
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    • pp.253-262
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    • 2001
  • Ligand-receptor clustering event is the most important step in leukocyte adhesion and spreading on endothelial cells. Intercellular adhesion molecule-1 (ICAM-1) has been shown to enhance leukocyte adhesion, but the molecular event during the process of adhesion is unclear. To visualize the dynamics of ICAM-1 movement during adhesion, we have engineered stable Chinese hamster ovary cell lines expressing ICAM-1 fused to a green fluorescent protein (IC1_GFP/CHO) and examined them under the fluorescence microscopy. The transfection of IC1_GFP alone in these cells was sufficient to support the adhesion of K562 cells that express $\alpha$L$\beta$2 (LFA-1) integrin stimulated by CBR LFA-1/2 mAb. This phenomenon was mediated by ICAM-1-LFA-1 interactions, as an mAb that specifically inhibits ICAM-1-LFA-1 interaction (RRl/l) completely abolished the adhesion of LFA-1* cells to IC1_ GFP/CHO cells. We found that the characteristic of adhesion was followed almost immediately (~10 min) by the rapid accumulation of ICAM-1 on CHO cells at a tight interface between the two cells. Interestingly, ICI_GFP/CHO cells projected plasma membrane and encircled approximately half surface of LFA-1+ cells, as defined by confocal microscopy. This unusual phenomenon was also confirmed on HUVEC after adhesion of LFA-1* cells. Neither cytochalasin D nor 2,3-butanedione 2-monoxime an inhibitor of myosin light chain kinase blocked LFA-1-mediated ICAM-1 clustering, indicating that actin cytoskeleton and myosin-dependent contractility are not necessary for ICAM-1 clustering. Taken together, we suggest that leukocyte adhesion to endothelial cells induces specialized form of ICAM-1 clustering that is distinct from immunological synapse mediated by T cell interaction with antigen presenting cells.

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요소회로 효소 유전자로 형질전환 된 Chinese Hamster Ovary 세포의 암모니아 제거능력과 세포성장률

  • Kim, Hong-Jin;Jeong, Myeong-Il;Jang, Yun-Jeong;Im, Mi-Hui;Kim, Ik-Hwan;Kim, Ik-Yeong
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.66-69
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    • 2001
  • Previously we developed a CHO cell line (CHO-OTC1-A19) expressing the first two enzymes of urea cycle. This cell line showed higher ammonia removal activity and faster growth rate than the vector controlled CHO cells (CHO-neo-5). The purpose of this study was to develop a cell line with higher ammonia removal activity than the cell line developed previously. To accomplish this, we constructed stable CHO cell lines expressing the first three, the first four, or all five enzymes of urea cycle by the stable transfection method. We finally selected CHO-AL-19 cell line expressing the first three, the first four enzymes of the cycle with higher ammonia activity than CHO-OTC1-A19 and CHO-n대-5 cell lines: 40% and 15% higher than those of CHO-neo-5 and CHO-OTC1-A19 cell lines 72 hour after culture started, respectively. It also showed 44% and 10% higher cell viability than CHO-neo-5 and CHO- OTC1-A19 cell lines at higher cell density. In addition, CHO-AL-19 cells showed 45%-60% and about 20% lower ammonia concentration per cell than those of CHO-neo-% and CHO-OTC1-A19 cell lines, respectively. These results indicate that CHO-AL-19 could be used in the production of human therapeutic proteins with higher efficiency.

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Highly Efficient Dye-Sensitized Solar Cells with Nonplatinized Graphene Oxide/Metal

  • Jeon, Yong-Seok;Lee, Dong-Uk;Kim, Jeong-U;Im, Jeong-Min;Seo, Seung-Hyeok;Han, Min-Su;Han, Chi-Hwan;Sin, Hyeon-Seok;Jeon, Yong-Seok
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2011.10a
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    • pp.11.2-11.2
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    • 2011
  • A key technological issue related to the implementation of dye-sensitized solar cells (DSSCs) is the replacement of Pt at the counter electrodes with an inexpensive and electro-chemically stable alternative. Carbon based nanomaterials could be promising candidates, but in practice they exhibit inadequate device performance. Here, we report very thin graphene oxide (GO)/metal hybrid films as transparent counter electrodes for high-efficiency DSSCs. Transparent GO/Pt and GO/Au hybrid films showed cell efficiencies of 9.2 and 9.0%, respectively (improvements of 9.5 and 7.1% over conventional Pt counter electrodes). More interestingly, highly stable DSSCs with GO hybrid films from relatively inexpensive metals such as Cu and Ni have been demonstrated with efficiency values comparable to Pt counter electrodes. The results reported in this study should enable low-cost fabrication of DSSCs because it allows the use of relatively inexpensive metals such as Au, Cu, Ni, and Ag that could not be previously employed in DSSCs with iodide/tri-iodide electrolyte due to corrosion.

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Studies on the Immobilization of Saccharomyces cerevisiae for Ethanol Production (효모의 Alginate 고정화에 관한 연구)

  • 한면수;하상도;정동효
    • Microbiology and Biotechnology Letters
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    • v.19 no.4
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    • pp.390-397
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    • 1991
  • Ethanol production by calcium alginate-immobilized baker's yeast (Saccharor/tyces cereviszae) was studied in the batch fermentation using glucose medium as a feed. Immobilied cells were stable between $30^{\circ}C$ and $40^{\circ}C$ whereas free cells were stable between $30^{\circ}C$ and $37^{\circ}C$ The beads were showed constant ethanol productivity during 720 hours (30 days) over. Fermentation characteristics of immobilized baker's yeast were examined changing the initial glucose concentration of broth in fermentation. Initial glucose concentrations employed were 50, 100, 150 and 200 g/l, respectively. In 15% gucose medium, maximum specific growth rate, maximum ethanol yield and ethanol concentration were observed as 0.092 $h^{-1}$, 0.45, 67.5 g/l, respectively.

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Advances in Materials for Proton Exchange Membrane based Fuel Cells

  • McGrath James E.
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.58-59
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    • 2006
  • Less than a decade ago, most alternate membrane materials for fuel cells relied upon a post-sulfonation process to generate ionic groups capable of transporting protons from the anode to the cathode. These random post sulfonations showed some promise, but in general they produced materials that were not sufficiently stable or protonically conductive at ion exchange capacities where aqueous swelling could be restricted. Our group began to synthesize disulfonated monomers that could be used to incorporate into random copolymer proton exchange membranes. The expected limitation was that the aromatic polymers might not be stable enough to withstand fuel cell conditions. However, this was mostly based upon an accelerated test known was the Fenton's Reagent Test, which did not seem to this author as being a reliable predictor of performance. A much better approach has been to evaluate the open circuit voltage (OCV) for alternate membranes, as well as the benchmark perfluorosulfonic acid systems. When this is done, the aromatic ionomers of this study, primarily based upon disulfonated polyarylene ether sulfones, show up quite well. Real time 3000 hours DMFC results have also been generated. Obtaining conductive materials at low humidities is another major issue where alternate membranes have not been particularly successful. In order to address this problem, multiblock copolymers with relatively high water diffusion coefficients have been designed, which show promise for conductivity at lowered humidity.

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