• Title/Summary/Keyword: GST

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Effects of Sardine Oil Fedding and Vitamin E Supplementation on Histopathological Changes and $\alpha$-L-Fucosidase Activity in Experimental Hepatocarcinogenesis (정어리유 섭취와 비타민 E 보충이 실험적 간 발암과정의 병리조직학적 변화와 $\alpha$-L-fucosidase의 활성도에 미치는 영향)

  • 김정희;윤혜진
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.1
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    • pp.119-126
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    • 2001
  • Effects of Sardine Oil Feeding and Vitamin E Supplementation on Histopathological Changes and $\alpha$-L-fucosidase activity in experimental hepatocarcinogenesis. Sprague-Dawley rats weighing 80~90 g were fed the diet containing either 15% corn oil (CO) or sardine oil (SO) with or without vitamin E supplements (dl-$\alpha$-tocopherol acetate 800 IU/kg diet) for 8 weeks. After 2 weeks of feeding, the rats were given a single intraperitoneal injectin of diethylnitrosamine (DEN, 200 mg/kg BW). From the fifth week, rats were given 0.02% acetylaminofluorene (AAF) in diet for 4 weeks. At the seventh week, 0.05% phenobarbital in liver and hepatic glutathione S-transferase palcental form positive (GST-P+) foci were examined by Hematoxylin& Eosin (H&E) staining and immunohistochemical method, respectively. Serum $\alpha$-L-fucosidase activity was determined. The livers fromt he carcinogen treated rats showed significantly increased formation of GST-P+ foci at sacrifice points while the livers fromthe non-carcinogen treated groups showed almost no foci. Although GST-P+ foci formation was not affected by dietary oil, it was increased unexpectedly by vitamin E supplementation. Histopathological changes were similar to patterns of GST-P+ foci formation in almost all groups. Serum $\alpha$-L-fucosidase activities were increased by carcinogen treatment in all dietary groups. $\alpha$-L-fucosidase activities were positively correlated with GST-P+ foci formation. There results suggest that excessive vitamin E supplementation can enhance hepatocarcinogenesis although the mechanisms involved are not clearly understood.

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Etching Characteristics of $Ge_2Sb_2Te_5$ Using High-Density Helicon Plasma for the Nonvolatile Phase Change Memory Applications (헬리콘 플라즈마를 이용한 $Ge_2Sb_2Te_5(GST)$ 상변화 재료의 식각 특성 검토)

  • Yoon, Sung-Min;Lee, Nam-Yeal;Ryu, Sang-Ouk;Shln, Woong-Chul;Yu, Byoung-Gon
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2004.11a
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    • pp.203-206
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    • 2004
  • For the realization of PRAM, $Ge_2Sb_2Te_5$ (GST) has been employed for the phase transition between the crystal and amorphous states by electrical joule heating. Although there has been a vast amount of results concerning the GST in material aspect for the laser-induced optical storage disc applications, the process-related issues of GST for the PRAM applications have not been reported. In this work, the etching behaviors of GST were investigated when the processing conditions were varied in the high-density helicon plasma. The etching parameters of RF main power, RF bias power, and chamber pressure were fixed at 600 W, 150 W, and 5 mTorr, respectively. For the etching processes, gas mixtures of $Ar/Cl_2$, $Ar/CF_4$, and $Ar/CHF_3$ were employed, in which the etching rates and etching selectivities of GST thin film in given gas mixtures were evaluated. From obtained results, it is found that we can arbitrarily design the etching process according to given cell structures and material combinations for PRAM cell fabrications.

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Effect of Cyclohexane Treatment on Serum Level of Glutathione S-Transferase Activity in Liver Damaged Rats ($CCl_4$ 에 의한 간손상 모델 실험동물에 있어서 cyclohexane 투여가 혈청 glutathione S-transferase 활성에 미치는 영향)

  • 오정대;윤종국
    • Journal of Environmental Health Sciences
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    • v.29 no.2
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    • pp.80-86
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    • 2003
  • To evaluate the effect of cyclohexane(CH) treatment on the serum levels of glutathion S-transferase(GST) activity in liver damaged animals, damaged liver was induced with pretreatment of 50% $CCl_4$ dissolved in olive oil (0.1 m1/100g body weight) intraperitoneally 17 times every other day. To $CCl_4$-treated rats, CH (1.56 g/kg body weight, i.p) was injected once and then the animals were sacrificed at 4 hours after injection of CH. The $CCl_4$-treated animals were identified as severe liver damage on the basis of liver functional findings, 1,e, increased serum levels of alanine aminotransferase(ALT), alkaline phosphate(ALP) and xanthine oxidase(XO) activities. On the other hand, $CCl_4$-treated animals injected with CH once($CCl_4$-pretreated animals) showed more decreased serum levels of ALT and XO, and more increased those of ALP rather than $CCl_4$-treated animals. In case of comparing the GST with ALT activity in liver, both $CCl_4$-treated and pretreated animals showed similar changing pattern of enzyme actvity. Especially $CCl_4$-pretreated animals showed significantly increased serum level of GST actvity compared with the $CCl_4$-treated those, whereas those of ALT showed reversed tendency. In aspects of GST enzyme kinetics, $CCl_4$-pretreated animals showed higher Vmax of liver GST enzyme than $CCl_4$-treated animals. In conclusion, injection of CH to the liver damaged rats led to enhanced liver damage and more increased activity of serum GST which may be chiefly caused by the enzyme induction.

Progesterone Effects on Microsomal Epoxide Hydrolase and Glutathione S-transferease mRNA Levels in Rats (랫드 간 Epoxide Hydrolase와 Glutathione S-Transferase 유전자 발현에 미치는 Progesterone의 효과)

  • Cho, Joo-Youn;Kim, Sang-Geon
    • The Korean Journal of Pharmacology
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    • v.32 no.2
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    • pp.233-241
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    • 1996
  • Previous studies have shown that glucocorticoid suppresses microsomal epoxide hydrolase(EH) gene expression and that EH expression is altered during pregnancy. The effects of progesterone on the expression of rat EH and certain glutathione S-transferase(GST) genes were examined in this study. Northern RNA blot analysis revealed that progesterone was effective in increasing hepatic EH mRNA levels at 12 h to 48 h after treatment with a maximal 9-fold increase being noted at 12 h time point. Nonetheless, multiple daily treatment with progesterone rather caused minimal relative increases in EH mRNA levels. GST Ya and Yb1/2 mRNA levels were also transiently elevated at 12 h after progesterone treatment, followed by gradual decreases from the maximal Increases at day 1, 2 and 5 post-treatment. These changes in EH and GST mRNA levels were noted only at a relatively high dose of progesterone. Furthermore, immunoblot analyses showed that rats treated with progesterone for 5 days failed to show EH or GST induction, indicating that progesterone-induced alterations in EH and GST mRNA levels do not reflect bona fide induction of the detoxifying enzymes. Concomitant progesterone treatment of rats with the known EH inducers including ketoconazole and clotrimazole failed to additively nor antagonistically alter EH mRNA levels. In contrast, dexamethasone substantially reduced ketoconazole- or clotrimazole-inducible EH expression. These results showed that progesterone stimulates the EH, GST Ya and Yb1/2 gene expression at early times followed by marked reduction in the RNA levels from the maximum after multiple treatment and that the changes in mRNA do not necessarily reflect induction of the proteins.

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Differential Effects of Nongenotoxic and Genotoxic Carcinogens on the Preneoplastic Lesions in the gat Liver

  • Kim, Dae-Joong;Lee, Kook-Kyung;Hong, Jin-Tae
    • Archives of Pharmacal Research
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    • v.21 no.4
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    • pp.363-369
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    • 1998
  • Glutathione S-transferase placental form (GST-P) positive foci development and its expression in liver exposed by nongenotoxic carcinogens phenobarbital (PB) and clofibrate (CF), and genotoxic carcinogen 2-amino-3-methylimidazo[4,5-f] quinoline (IQ) were investigated as a measure of carcinogenic potential of these chemicals. Male F344 rats were initially given a single intraperitioneal injection of diethyinitrosamine (200 mg/kg), and 2 weeks later, animals were fed diets containing 0.03% IQ or 0.5% CF or 0.05% PB or basal diet as a control for 6 weeks. All rats were subjected to two-thirds partial hepatectomy (PH) at week 3. Sequential sacrifice of rats was performed at 8 weeks or 52 weeks, and liver tissues were examined for immunohistochemical staining of GST-P positive foci. The numbers (No./$cm^2$) and areas ($mm^2$/ $cm^2$) of GST-P positive foci were increased by IQ or PB, but were decreased by CF compare to the control. Consistent with the development of GST-P positive foci, a time-related increase in the expression of GST-P mRNA was found in the rats treated with IQ, whereas CF decreased it. The incidence of hepatocellular carcinoma at 52 weeks was increased by all three chemicals. These results show that PB and IQ induced GST-P positive foci, but the peroxisome proliferator CF did not, which suggest that the prediction of carcinogenic potency based on the development of prenoplastic foci may cause false negative in a particular category compounds like peroxisome proliferators.

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Caspase-3-facilitated Stoichiometric Cleavage of a Large Recombinant Polyprotein (카스파제-3 효소를 이용한 폴리-단백질의 정량적 프로세싱 분석)

  • Kim, Moonil
    • Journal of Life Science
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    • v.25 no.4
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    • pp.385-389
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    • 2015
  • In this study, it is reported that a large polyprotein can be stoichiometrically cleaved by the use of caspase-3-dependent proteolysis. Previously, it has been shown that the proteolytic IETD motif was partially processed when treated with caspase-3, while the DEVD motif was completely cleaved. The cleavage efficiency of the DEVD-based substrate was approximately 2.0 times higher than that of the IETD substrate, in response to caspase-3. Based on this, 3 protein genes of interest were genetically linked to each other by adding two proteolytic cleavage sequences, DEVD and IETD, for caspase-3. Particularly, glutathione-S transferase (GST), maltose binding protein (MBP), and red fluorescent protein (RFP) were chosen as model proteins due to the variation in their size. The expressed polyprotein was purified by immobilized metal ion affinity chromatography (IMAC) via a hexa-histidine tag at the C-terminal end, showing 93 kDa of a chimeric GST:MBP:RFP fusion protein. In response to caspase-3, cleavage products, such as MBP:RFP (68 kDa), MBP (42 kDa), RFP (26 kDa), and GST (25 kDa), were separated from a large precursor GST:MBP:RFP (93 kDa) via SDS-PAGE. The results obtained from this study indicate that a multi-protein can be stoichiometrically produced from a large poly-protein by using proteolytic recognition motifs, such as DEVD and IETD tetra-peptides, for caspase-3.

Investigation of Ge2Sb2Te5 Etching Damage by Halogen Plasmas (할로겐 플라즈마에 의한 Ge2Sb2Te5 식각 데미지 연구)

  • Jang, Yun Chang;Yoo, Chan Young;Ryu, Sangwon;Kwon, Ji Won;Kim, Gon Ho
    • Journal of the Semiconductor & Display Technology
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    • v.18 no.4
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    • pp.35-39
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    • 2019
  • Effect of Ge2Sb2Te5 (GST) chalcogen composition on plasma induced damage was investigated by using Ar ions and F radicals. Experiments were carried out with three different modes; the physical etching, the chemical etching, and the ion-enhanced chemical etching mode. For the physical etching by Ar ions, the sputtering yield was obtained according to ion bombarding energy and there was no change in GST composition ratio. In the plasma mode, the lowest etch rate was measured at the same applied power and there was also no plasma induced damage. In the ion-enhanced chemical etching conditions irradiated with high energy ions and F halogen radicals, the GST composition ratio was changed according to the density of F radicals, resulting in higher roughness of the etched surface. The change of GST composition ratio in halogen plasma is caused by the volatility difference of GST-halogen compounds with high energy ions over than the activation energy of surface reactions.

A Study of the Effectiveness of Hwanggeum-tang and Gamchosasim-tang in the Mice Model of Inflammatory Bowel Disease (염증성 장 질환 동물 모델에서 황금탕과 감초사심탕의 효능 연구)

  • Kim, Young-kwang;Moon, Young-ho
    • The Journal of Internal Korean Medicine
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    • v.42 no.3
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    • pp.351-374
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    • 2021
  • Objectives: This study investigates the mechanism of Hwanggeum-tang (HGT) and Gamchosasim-tang (GST) on inflammatory bowel disease (IBD). Methods: The mice (C57BL/6N) were treated with distilled water and 3% dextran sulfate sodium (DSS) to experimentally induce ulcerative colitis. The mice were divided into 7 groups of (6 mice: normal, negative control, positive control (with sulfasalazine), 4 experimental groups (with HGT and GST, respectively). RAW 264.7 cells were used for cell experiments. The experiment was conducted in two ways: in vitro and in vivo. Results: In the experimental group (HGT, GST) of in vitro experiments, NO production decreased, and significant changes in gene expression and protein activation were observed. The length of the colon recovered in the experimental groups (HGT, GST) of the in vivo experiment was longer than that of the negative control group, and the mucosal barrier was recovered. Sone significant changes in the amount of mRNA expression were partially observed, and significant changes in protein activation also were confirmed. Conclusions: HGT and GST are effective in treating IBD caused by DSS. In the same herbal preparation group, the higher the concentration, the better the experimental effect, and when the same concentration was tested, HGT was more effective than GST. Herbal medicine has a higher antioxidant effect than sulfasalazine, so it is also excellent for cell protection.

Analysis on the substrate specificity and inhibition effect of Brassica oleracea glutathione S-Transferase (양배추 유래의 글루타티온 전달효소의 기질 특이성 및 저해 효과 분석)

  • Park, Hee-Joong;Lee, Hee-Jin;Kong, Kwang-Hoon
    • Analytical Science and Technology
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    • v.22 no.3
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    • pp.228-234
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    • 2009
  • To gain further insight into herbicide detoxification of plant, we purified a glutathione S-transferase from Brassica oleracea (BoGST) and studied its substrate specificity towards several xenobiotic compounds. The BoGST was purified to electrophoretic homogeneity with approximately 10% activity yield by DEAE-Sephacel and GSHSepharose column chromatography. The molecular weight of the BoGST was determined to be approximately 23,000 by SDS-polyacrylamide gel electrophoresis and 48,000 by gel chromatography, indicating a homodimeric structure. The activity of the BoGST was significantly inhibited by S-hexyl-GSH and S-(2,4-dinitrophenyl)GSH. The substrate specificity of the BoGST displayed high activities towards CDNB, a general GST substrate and ethacrynic acid. It also exhibited GSH peroxidase activity toward cumene hydroperoxide.

Inhibitory Effort of the N-terminal GST on the Tautomerase Activity of Macrophage Migration Inhibitory Factor (GST 융합 시스템에서 나타나는 macrophage migration inhibitory factor의 tautomerase 활성 저해에 관한 연구)

  • Kim Sang-Soo;Kim Kyung-Hee;Park Hyo-Jin;Hur Eun-hye;Rhim Hyangshuk
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.961-967
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    • 2005
  • Macrophage migration inhibitory fartor (MIF), known as a cytokine, is a multifunctional protein that is ubiquitously expressed in a variety of cells and tissues; however, enzymatic function of MIF still remains elusive in cells. In this study, we assessed details of the tautomerase activity of MIF. We established rapid purification condition for MIF by using pGEX system and compared the L-dopachrome tautomerase activity of GST-MIF, tMIF, and MIF. The results show that GST (glutathione S-transferase)-epitope tag or N-terminal amino acids flanking the essential $P^{2}$ almost completely abrogated L-dopachrome tautomerase activity of MIF. Subsequently, to determine whether the N-terminal tags have effects on oligomerization of MIF, protein cross-linking products were analyzed on $15\%$ SDS-PACE. The result demonstrates that N-terminal tags are dispensable for the formation of MIF's homooligomers. Thus, the results imply that exposure of If containing hydrophobic pocket in the active site is critical for L-dopachrome tautomerase activity of MIF. In addition, our study suggest that the MIF's tautomerase activity might be influenced by interacting with cellular partners.