• Title/Summary/Keyword: HEP system

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Anti-cancer and anti-inflammatory effects of convergence of ginsenoside Rh2, compound K isolated from amplified red ginseng (증폭시킨 홍삼으로부터 분리한 ginsenoside Rh2, compound K의 융복합적 항암 및 항염효과)

  • Kim, Young-Ho;Kim, Jong-Du
    • Journal of Digital Convergence
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    • v.15 no.11
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    • pp.285-295
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    • 2017
  • This study aims to provide basic data on useful functional ingredients in red ginseng by studying the anti-inflammatory and anti-cancer effects of convergence of ginsenoside Rh2(Rh2) and compound K(CK) isolated from amplified red ginseng. Therefore we examined cytotoxicity in Hep3B, activity of IL-6 induced STAT3 luciferase and survival concentration of cells in B16F10 and HaCa T. According to the experimental results, when the Rh2 and CK mixture were 10 ug/ml, there was no cytotoxicity in Hep3B cells and the anti-inflammatory effect of IL-6 reduction ratio was 102%. In addition, Rh2 and CK mixture were observed to be toxic in melanoma cell line B16F10 and HaCa T (human keratinocyte) at 50 uM. FACS(fluorescence activated cell sorting) analysis showed that annexin V was not expressed and melanoma cells and keratinocyte were desorbed and killed. It can be assumed that the mechanism of killing through this phenomenon is due to the cell death of anoikis-type, and it is necessary to study the changes of cell adhesion proteins in the future in order to clarify the cell death signal system.

A Study on the Inhibition of 2-deoxy-D-Glucose Transport of the Endogenous Glucose Transporters in Spodoptera frugiperda Clone 21-AE Cells by Using Hexoses

  • Lee Chong-Kee
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.487-492
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    • 2005
  • The baculovirus/insect cell expression system is of great value in the study of structure-function relationships in mammalian glucose-transport proteins by site-directed mutagenesis and for the large-scale production of these proteins for mechanistic and biochemical studies. Spodoptera frugiperda Clone 21 (Sf2l) cells grow well on TC-100 medium that contains $0.1\%$ D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, very little is known about the properties of the endogenous sugar transporter(s) in Sf2l cells, although a saturable transport system for hexose uptake has been previously revealed in the Sf cells. In order to further examine the substrate and inhibitor recognition properties of the Sf2l cell transporter, the ability of hexoses to inhibit 2-deoxy-D-glucose (2dGlc) transport was investigated by measuring inhibition constants $(K_i)$. The $K_i's$ for reversible inhibitors were determined from plots of uptake versus inhibitor concentration. Transport was effectively inhibited by D-mannose and D-glucose. Of the hexoses tested, L-glucose had the least effect on 2dGlc transport in the Sf2l cells, indicating that the transport is stereoselective. Unlike the human HepG2 type glucose transport system, D-mannose had a somewhat greater affinity for the Sf2l cell transporter than D-glucose, implying that the hydroxyl group at the C-2 position is not necessary for strong binding. However, epimerization at the C-4 position of D-glucose (D-galactose) resulted in a dramatic decrease in affinity of the hexose for the Sf2l cell transporter. Such a lowering of affinity might be the result of the involvement of the C-4 hydroxyl in hydrogen bonding. It is therefore suggested that Sf2l cells were found to contain an endogenous sugar transport activity that in several aspects resembles the human HepG2 type glucose transporter, although the insect and human transporters do differ in their affinity for cytochalasin B.

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Inhibitory Effect of Methanol Extracts and Solvent Fractions from Meju on Mutagenicity and Growth of Human Cancer Cells (메주 메탄올 추출물 및 분획물의 항돌연변이 및 인체 암세포 성장 억제 효과)

  • Lim, Sun-Young;Park, Kun-Young;Lee, Sook-Hee;Choi, Jae-Soo
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.76-81
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    • 2007
  • Inhibitory effects of methanol extracts and several solvent fractions from meju on mutagenicity in vitro genotoxicity (SOS chromotest) and growth of human cancer cells (AGS gastric adenocarcinoma and Hep 3B hepatocellular cancinoma cells) were studied. The treatment of meju methanol extracts $(100{\mu}g/assay)$ to SOS chromotest system inhibited N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) induced mutagenicity by 36%. However, the ethylacetate and dichloromethane fractions from meju methanol extracts showed the stronger antimutagenic effects (91% and 91%, respectively) in SOS chromotest. In sulforhodamine B (SRB) assay, the treatments of ethylacetate and dichloromethane fractions (2 mg/assay) significantly inhibited the growth of AGS and Hep 3B cancer cells by 64% and 71%, respectively. These results indicated that meju had inhibitor)r effects on MNNG in SOS mutagenic system and growth of human cancer cells, suggesting that its antimutagenic effect may be relative to activity of doenjang.

An Embodiment of High Energy Physics Data Grid System (고에너지물리 데이타 그리드 시스템의 구현)

  • Cho Ki-Hyeon;Han Dae-Hee;Kwon Ki-Hwan;Kim Jin-Cheol;Yang Yu-Chul;Oh Young-Do;Kong Dae-Jung;Suh Jun-Suhk;Kim Dong-Hee;Son Dong-Chul
    • Journal of KIISE:Computer Systems and Theory
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    • v.33 no.7
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    • pp.390-398
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    • 2006
  • The objective of the High Energy Physics(HEP) is to understand the basic properties of elementary particles and their interactions. The CMS(Compact Muon Solenoid) experiment at CERN which will produce a few PetaByte of data and the size of collaboration is around 2000 physicists. We cannot process the amount of data by current concept of computing. Therefore, an area of High Energy Physics uses a concept of Tier and Data Grid. We also apply Data Grid to current High Energy Physics experiments. In this paper, we report High Energy Physics Data Grid System as an application of Grid.

IN VITRO DRUG METABOLISM BY A HUMAN P450 EXPRESSION SYSTEM USING VACCINIA VIRUS.

  • S.Ono;Ha, T.tanaka;H.Hotta;M.Tsutsui;F.J.Gonzalez;T.Aoyama;T.Satoh
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.153-163
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    • 1994
  • P450s are a superfamily of heme-containing monooxygenases and important in the metabolism of numerous physiological substrates and foreign compounds. It has been established that tilers are at least 30 distinct human isoforms of P450. Four families containing numerous individual P450s are mainly responsible for metabolizing foreign compounds, A cDNA expression system in which individual human P450s are synthesized in cultured human hepatoma (Hep G2) cells infected with a recombinant vaccinia virus containing human P450 cDNA has been constructed.

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The antioxidative effects of Ampelopsis brevipedunculata extracts (개머루덩굴 추출물의 항산화 효과)

  • Rhim, Tae-Jin;Choi, Moo-Young
    • Korean Journal of Plant Resources
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    • v.23 no.5
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    • pp.445-450
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    • 2010
  • This study was carried out to investigate the antioxidative capacity of Ampelopsis brevipedunculata 95% ethanol extracts. The concentration of A. brevipedunculata extract at which DPPH radical scavenging activity was inhibited by 50% was 0.42 mg/mL as compared to 100% by pyrogallol as a reference. Total antioxidant status was examined by total antioxidant capacity against ABTS radical reactions. Total antioxidant capacities of A. brevipedunculata extract at the concentrations of 0.1 and 1 mg/mL were 0.65 and 3.71 mM Trolox equivalents, respectively. Oxygen radical absorbance capacities of A. brevipedunculata extract at the concentrations of 5 and $100\;{\mu}g/mL$ were 22.75 and $131.25\;{\mu}M$ gallic acid equivalents, respectively. Superoxide scavenging activities of A. brevipedunculata extract at the concentrations of 0.1 and 1 mg/mL were 27.7 and 56.0%, respectively. Total phenolic contents of A. brevipedunculata extract at the concentrations of 0.5 and 2.0 mg/mL were 0.55 and 2.06 mM gallic acid equivalents, respectively. A. brevipedunculata extract at the concentration of 0.1 mg/mL inhibited 0.2 mM and 0.5 mM tert-butyl hydroperoxide induced cyototoxicity by 36.2 and 23.3%, respectively, in HepG2 cell culture system. Thus strong antioxidant and cytotoxicity-inhibiting effects of A. brevipedunculata extract seem to be due to, at least in part, the prevention from free radicals-induced oxidation as well as high levels in total phenolic contents.

Antioxidant and Anticancer Activities of Ginseng Treated with Traditional Rice Wine Steam Process Method (전통적 탁주증자법으로 처리한 홍삼의 일부 항산화 및 항암효과)

  • Ye, Eun-Ju;Kim, Soo-Jung;Park, Chang-Ho;Bae, Man-Jong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.5
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    • pp.599-604
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    • 2005
  • The aim of this study was to develop the new processing method for ginseng. To investigate the efficacy of the new product (the traditional rice wine steamed-red ginseng: RWS-RGS), antioxidant and anticancer effects of RWS-RGS were examined. The DPPH radical scavenging effect of RWS-RGS extracted with ethanol was increased in dose-dependent manner Especially, A3 ($3^{rd}$ traditional rice wine steamed-red ginsengs) exhibited effective DPPH radical scavenging activity. Nitrite scavenging effect of white ginseng (W.G), red ginseng (R.G) and RWS-RGS ($A1\~A9:\;1^{st}$ traditional rice wine steamed-red $ginseng\~9^{th}$ traditional rice wine steamed-red ginseng) were $25.9{\pm}4.4\%,\;12.9{\pm}1.1\%\;and\;26.2{\pm}0.1\~56.1{\pm}0.6\%$ at pH 1.2, respectively. The antitumor effects of W.G, R.G and RWS-RGS (A9) were examined in Hep3B cancer cells. Their growth inhibition against Hep3B cancer cells showed $19.6{\pm}4.5\%,\;54.5{\pm}6.1\%,\;96.3{\pm}2.4\%$ at 5,000 ppm, respectively. These result suggest that the traditional rice wine steamed ginseng will be useful product with antioxidant and antitumor effect.

Antimutagenicity and Anticancer Activity of Soybean Fractions Extracted by Solvents (대두 분획물의 항돌연변이 및 항암활성 효과)

  • Lim, Sun-Young
    • Journal of Life Science
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    • v.17 no.10
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    • pp.1368-1373
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    • 2007
  • Inhibitory effects of several solvent fractions from soybean on mutagenicity using Salmonella typhimurium TA 100 in Ames test and growth of human cancer cells (AGS gastric adenocarcinoma, Hep 3B hepatocellular cancinoma and HT-29 colon cancer cells) were studied. The treatment of dichloromethane and ethylacetate fractions (2.5 mg/assay) extracted from soybean to Ames test system inhibited aflatoxin $B_1\;(AFB_1)$ induced mutagenicity by 83%, respectively, and showed a higher antimutagenic effect than other solvent fractions. In case of N-methyl-N#-nitro-N-nitrosoguamidine (MNNG) induced mutagenicity, the ethylacetate fraction showed the highest inhibitory effect (by 67%) among solvent extracts, although the inhibitory effect was not stronger compared with $AFB_1$ induced mutagenicity. In sulforhodamine B (SRB) assay, the treatment of ethylacetate fraction (2 mg/assay) significantly inhibited the growth of AGS, Hep 3B and HT-29 cancer cells by 66%, 73% and 77%, respectively, followed with the intermediate and dichloromethane fractions. These results indicated that soybean fraction extracted with ethylacetate had higher inhibitory effects on $AFB_1$ and MNNG in Ames test and growth inhibition activity to human cancer cells was appeared, suggesting that soybean fraction extracted with ethylacetate may contain the biologically active compounds.

Expression of Polyhistidine-Containing Fusion Human HepG2 Type Glucose Transport Protein in Spodoptera Cells and Its Purification Using a Metal Affinity Chromatography

  • Lee, Chong-Kee
    • Biomedical Science Letters
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    • v.16 no.3
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    • pp.201-206
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    • 2010
  • In order to develop procedures for the rapid isolation of recombinant sugar transporter in functional form from away from the endogenous insect cell transporter, gene fusion techniques were exploited. Briefly, BamH1-digested human HepG2 type glucose transport protein cDNA was first cloned into a transfer vector pBlueBacHis, containing a tract of six histidine residues. Recombinant baculoviruses including the human cDNA were then generated by allelic exchange following transfection of insect cells with wild-type BaculoGold virus DNA and the recombinant transfer vector. Plaque assay was then performed to obtain and purify recombinant viruses expressing the human transport protein. All the cell samples that had been infected with viruses from the several blue plaques exhibited a positive reaction in the immnuassay, demonstrating expression of the glucose transport protein. In contrast, no color development in the immunoassay was observed for cells infected with the wild-type virus or no virus. Immunoblot analysis showed that a major immunoreactive band of apparent Mr 43,000~44,000 was evident in the lysate from cells infected with the recombinant baculovirus. Following expression of the recombinant fusion protein with the metal-binding domain and enterokinase cleavage site, the fusion protein was recovered by competition with imidizole using immobilized metal charged resin. The leader peptide was then removed from the fusion protein by cleavage with porcine enterokinase. Final separation of the recombinant protein of the interest was achieved by passage over $Ni^{2+}$-charged resin under binding conditions. The expressed transport protein bound cytochalasin B and demonstrated a functional similarity to its human counterpart.

Studies on the Possible Mechanisms of Protective Activity Against $\alpha$-Amanitin Poisoning by Aucubin

  • Lee, Dong-Hee;Cho, In-Goo;Park, Moon-Soo;Kim, Ki-Nam;Chang, Il-Moo;Mar, Woong-chon
    • Archives of Pharmacal Research
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    • v.24 no.1
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    • pp.55-63
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    • 2001
  • Aucubin, an irdoid g1ucoside, was investigated to determine whether it has a stimulating effect on $\alpha$-amanitin excretion in $\alpha$-amanitin intoxicated rats, and whether there is binding activity to calf thymus DNA. High-performance liquid chromatography (HPLC) analysis of $\alpha$-amanitin in rat urine allowed quantitative measurement of the $\alpha$-amanitin concentration with a detection limit of 50${mu}g/ml$. In this system, a group treated with both $\alpha$-amanitin and aucubin showed that o(-amanitin was excreted about 1.4 times faster than in the $\alpha$-amanitin only treated group. Our previous results showed that the toxicity of $\alpha$-amanitin is due to specific inhibition of RNA polymerase activity and the resultant blockage of the synthesis of certain RNA species in the nucleus. However, no significant activity change on RNA polymerase from Hep G2 cells was observed when aucubin was treated with $\alpha$-amanitin at any concentration tested. Nevertheless, aucubigenin inhibited both DNA polymerase (IC50, 80.5${mu}g/ml$) and RNA polymerase (IC50, 135.0${mu}g/ml$) from the Hep G2 cells. The potential of both $\alpha$-amanitin and aucubin to interact with DNA were examined by spectrophotometric analysis. $\alpha$-Amanitin showed no significant binding capacity to calf thymus DNA, but aucubin was found to interact with DNA, and the apparent binding constant ($K_{app}$) and apparent number of binding sites per D7A phosphate ($B_{app}$) were 0.45$0.45{\times}$$10^4$ $M^{-1}$ and 1.25, respectively.

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