• Journal of the Korean Magnetic Resonance Society
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• v.24 no.3
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• pp.77-85
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• 2020

Stable isotope enrichment in proteins is necessary for high-resolution nuclear magnetic resonance (NMR) experiments. Although methods for ¹³C, ¹⁵N and ²H-enrichment in prokaryotic cells are well established, full processing and correct folding of complex protein systems require higher organisms as the expression host. In the present study, we review recent efforts to enrich stable isotopes in mammalian cells for protein NMR studies.

• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.115-118
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• 2000

Recombinant plasmids harboring a heterologous gene coding for the enhanced green fluorescent protein (EGFP) were transfected and expressed in Drosophila melanogaster S2 cells. A stable transformation of polyclonal cell populations expressing EGFP were isolated after 4 weeks of selection with hygromycin B. The recombinant EFGP expressed in transformed S2 cells consisted of a molecular weight of 27 kDa. EGFP expression was also confirmed by fluorometric measurement. The maximum EGFP concentration was about 9.3 mg/I. The present findings demonstrate not only the successful stable expression of EGFP in Drosophuila was about 9.3 mgI. The present findings demonstrate not only the successful stable expression of EGFP in Drosophila S2 cells, but also the use of EGFP as a reporter to analyze gene expression, with its potential of a Drosophila cell expression system for recombinant protein production being an alternative to a baculovirus-insect cell expression system.

• BMB Reports
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• v.29 no.4
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• pp.286-293
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• 1996

Association of antigenic peptide with class I MHC is believed to be crucial for maintaining stable conformation of class I molecules. T2 cells that are defective in TAP gene function mainly express class I molecules with an unstable conformation due to little or no association with antigenic peptides, whereas T1 cells that are normal in TAP gene function mainly express the stable form of class I molecules. In this work, attempts were made to determine the molecular stability of stable and unstable class I molecules. Dissociation of HLA-A2 molecules on T1 and T2 cells was monitored by flow cytometry using anti-HLA-A2 antibody after the cells were treated with brefeldin A to shut down the transport of newly-assembled HLA-A2. Estimated dissociation rate constants for the stable and unstable forms of HLA-A2 were 0.076 $h^{-1}$ and 0.66 $h^{-1}$, respectively. It appeared that both T1 and T2 cells express stable and unstable class I complex, but with different ratios of the two forms. Furthermore, $interferon-{\gamma}$ treatment of T1 cells appeared to induce the expression of both the stable and unstable class I molecules. These results demonstrate that class I MHC molecules can be divided into two groups in terms of structural stability and that they exist on the cell surface in both forms in a certain ratio.

• Journal of Microbiology and Biotechnology
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• v.29 no.1
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• pp.55-58
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• 2019

Development of stable rCHO cell lines is still time consuming and labor intensive, although it is a critical step in the commercial development of recombinant antibodies. The current work demonstrates, for the first time, that electroporation of CHO cells with DMSO can enhance stable expression of recombinant antibodies in rCHO cells. Electroporation with DMSO resulted in an average 3.7-fold and 2.8-fold increases in expression levels of aflibercept and pembrolizumab, respectively, in pools of stable rCHO cells. It also resulted in an average of 2.2-fold and 2.6-fold increases in the expression of aflibercept and pembrolizumab, respectively, in single-cell derived rCHO clones. Simple batch cultures of rCHO cell clones with the highest expression produced 1.0 g/l for aflibercept and 1.4 g/l for pembrolizumab without a time-consuming gene amplification process. Electroporation with DMSO also shortened the development of rCHO cell lines to 2-3 months, allowing rapid establishment of stable rCHO cell lines with a desirable expression level antibodies.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 2004.07a
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• pp.169-172
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• 2004

The effect of the process parameters on the stable lifetime in rapid thermal firing(RTF) was investigated in order to optimize the process for the Cz-silicon. The process temperature was varied between $700^{\circ}C\;and\;950^{\circ}C$ while the process time was chosen 1 s and 10 s. At below $850^{\circ}C$ the stable lifetime for 10 s is higher than that for 1 s and increases with increasing by the process temperature. However, at over $850^{\circ}C$ the improved stable lifetime is not dependent on the process time and temperature. On the other hand, two high temperature processes in solar cell fabrics are combined with the optimized process and the non-optimized process. The last process determines the stable lifetime. Also, the degraded stable lifetime could be increased by processing in optimized process. The decreased lifetime can increase using the optimized oxidation process, which is a final process in solar cells. Finally, the optimized and non-optimized processes are applied solar cells.

• The Journal of the Korean Society for Microbiology
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• v.3 no.1
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• pp.51-65
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• 1968

The site of the synthesis of Japanese encephalitis virus(JEV) in the actinomycin-treated and infecter PS Y15 cells(a porcine kidney stable cell line) was observed by the immunofluorescent antibody technique, acridine orange staining, and the autoradiographic analysis. In the parallel studies by immunofluorescent technique and acridine orange staining it the infected cells, Viral protein(as an antigen) and viral RNA were detected at the same site of cytoplasm. In the autoradiographic analysis, the cytoplasmic labeling of $^3H$-uridine was due to the synthesis of JEV-RNA, while the nucleolus and nucleus were not involved. In the autoradiographic studies on the secton of infected cells, the $^3H$-uridine was frequently incorporated around the cytoplasmic vacuoles. This localization of labeling agreed with the site of acridine orange positive granules. The results suggest that the syntheses of the viral RNA and viral protein occurred in the similar site of cytoplasm of the infected cells, and also the virus particles seem to be assembled in the sites of the viral RNA and protein syntheses.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.2
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• pp.607-611
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• 2012

Non-toxic stimulation of dendritic cells (DCs), which are central immunomodulators, may aid the prevention of cancer. Furthermore, induction of apoptosis in cancer cells by anticancer agents contributes to the induction of DC maturation. We previously reported that extracts from $Pinus$ $parviflora$ Sieb. et Zucc pine cone and $Mucuna$ seed induce differentiation of mouse bone marrow cells into mature dendritic cells and also induce apoptosis in various human cancer cell lines. In the present study, we screened 31 kinds of edible beans with biological activity similar to that of extracts from pine cone and $Mucuna$ and found that the heat-stable extract from azuki bean ($Vigna$ $angula$) stimulated differentiation of bone marrow cells into immature DCs with the greatest efficacy. The level of IL-6 produced by sequential treatment of DCs with azuki extract and lipopolysaccharide was the highest among the examined beans. Azuki extract also inhibited the growth of human leukemia U937 cells, leading to induction of apoptosis. These results suggest that azuki bean and its extract are immunopotentiating foods that can be used as a dietary supplement for cancer prevention and immunotherapy.

• Journal of the Korean Mathematical Society
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• v.42 no.5
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• pp.1071-1086
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• 2005

In this paper, we introduce a discrete time to the model to describe the time between infection of a CD4$^{+}$ T-cells, and the emission of viral particles on a cellular level. We study the effect of the time delay on the stability of the endemically infected equilibrium, criteria are given to ensure that the infected equilibrium is asymptotically stable for all delay. We also obtain the condition for existence of an orbitally asymptotically stable periodic solution.

• Proceedings of the Korean Society of Toxicology Conference
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• 2001.10a
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• pp.202-202
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• 2001

Stable MCF-7-ERE cells, in which pERE-Luc reporter gene has been stably integrated into the genome of the MCF-7 cells, were used to detect the estrogenic activity of various dietary flavonoids.in either pure chemical or mixtures. Estradiol (E2) induced luciferase activity in dose dependent manner and this activity was inhibited by tamoxifen (Tam) concomitant treatment.(omitted)

• The Transactions of The Korean Institute of Electrical Engineers
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• v.63 no.12
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• pp.1671-1674
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• 2014

This paper presents a frequency-dependent cell impedance analysis chip for use in cancer and normal cell discrimination. The previous cell impedance analysis chips for flowing cells cannot allow enough time for cell-to-electrode contact to monitor frequency-dependent impedance response. Another type of the previous cell impedance analysis chips for the cells clamped by membranes need complex sample control for making stable cell-to-electrode contact. We present a new impedance analysis chip using the microchamber array, on which a PDMS cover is placed to make stable cell-to-electrode contact for the individual cell trapped in each microchamber; thus achieving frequency-dependent single-cell impedance analysis without complex sample control. Compared to the normal cells, the magnitude of NHBE cells is $60.07{\sim}97.41k{\Omega}$ higher than A549 cells in the frequency range of 95.6 kHz~2MHz and the phase of NHBE is $3.96^{\circ}{\sim}20.8^{\circ}$ higher than A549 cells in the frequency range of 4.37 kHz~2MHz, respectively. It is demonstrated experimentally that the impedance analysis chip performs frequency-dependent cell impedance analysis by making stable cell-to-electrode contact with simple sample control; thereby applicable to the normal cell and cancer cell discrimination.