• Title/Summary/Keyword: GST-P

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Effect of Turbid Water on Fishes in the Streams of Imha Reservoir (임하호 유입지천에 서식하는 어류에 미치는 탁수의 영향)

  • Yu, Sam-Hwan;Kim, Jeong-Sook;Shin, Myung-Ja;Lee, Jong-Eun;Seo, Eul-Won
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1410-1416
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    • 2009
  • The present study aims to examine the effect of turbid water on fishes in streams which branch into a turbid water area (Yeongyang-gun) and a non-turbid water area (Cheongsong-gun), and finally flow into the Imha reservoir. In a comparison of water quality, the chemical status of the water showed higher pH, DO and SS in the turbid water area than in the non-turbid water area. Also, high density of clay minerals such as vermiculite (V) and illite (I), which is from clay mineral leakage during rainfall, was detected in turbid water, resulting in an increase of turbidity. Fishes inhabiting the turbid water showed irregular spaces in gill lamella, cell separation, edema, and clubbing in epithelial tissues. Also, the gill surface showed roughness and plenty of muddy debris substances inside the gills. The Bowman's space was expanded because of contraction of the glomerulus in the Bowman's space of the kidney tissues. Antioxidant enzymes such as SOD, CAT, GPX, and GST showed higher activities in the specific tissues, muscles and kidney, of fishes living in turbid water than in the non-turbid area. We suggested that; first, the antioxidant activities were increased due to removal of harmful radicals generated in fish bodies in the turbid water area, second, long-time exposure of these histological changes in the tissues might have induced secondary lesion accompanying the inaccurate physiological constancy of fishes.

Antitumor Effect of Young Radish Kimchi Prepared with Young Radish Cultivated in the Soil Containing Sulfur on Sarcoma-180 Tumor Cells Transplanted Mice (유황처리 열무로 제조한 열무김치의 Sarcoma-180 암세포에 대한 고형암 성장 억제효과)

  • Kong, Chang-Suk;Bak, Soon-Sun;Rhee, Sook-Hee;Kil, Jeung-Ha;Rho, Chi-Woong;Hwang, Hae-Jun;Kim, Nak-Ku;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.10
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    • pp.1520-1524
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    • 2005
  • Young radishes (YR, yeolmu in Korean) were cultivated in the soil with and without sulfur. Young radish kimchi-general (YR kimchi-G) was prepared with YR commonly cultivated in the soil without sulfur. Young radish kimchl-sulfur (YR kimchi-S) and young radish kimchi-sulfur with lime mortar (YR kimchi-SL) were prepared with the YR cultivated in the soil with sulfur an4 sulfur added lime mortar on it respectively. Antitumor effects of methanol extracts from the YR kimchis were investigated in sarcoma-180 tumor cell transplanted mice. The solid tumor growth was significantly inhibited by the YR kimchi-SL prepared with YR grown in the soil with sulfur (p < 0.05). The treatment of the methanol extracts from YR kimchi-S and -SL increased the glutathione S-transferase activities and glutathione contents in the livers, compared to thlose of YR kimchi-G and the control. One of the antitumor effects by the YR kimchis was due to the increased the glutathione levels and the glutathione S-transferase activity which is phase 2 enzyme. These results also suggested that the antitumor effects of YR kimchi can be enhanced by using YR cultivated differently in the presence of sulfur that can help to produce sulfur-containing compounds in YR.

Expression of Human SOD1 and Mutant SOD1 (G93A) in E. coli and Identification of SOD1 as a Substrate of HtrA2 Serine Protease (대장균에서의 human SOD1과 mutant SOD1 (G93A) 단백질의 발현과 HtrA2의 기질 여부 확인에 관한 연구)

  • Kim, Goo-Young;Kim, Sang-Soo;Park, Hyo-Jin;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.5
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    • pp.716-722
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    • 2006
  • Superoxide dismutase (SOD) is physiologically important in regulating cellular homeostasis and apoptotic cell death, and its mutations are the cause of familial amyotrophic lateral sclerosis (FALS). Mitochondrial serine protease HtrA2 has a pro-apoptotic function and has known to be associated with neurodegenerative disorders. To investigate the relationship between genes associated with apoptotic cell death, such as HtrA2 and SOD1, we utilized the pGEX expression system to develop a simple and rapid method for purifying wild-type and ALS-associated mutant SOD1 proteins in a suitable form for biochemical studies. We purified SOD1 and SOD1 (G93A) proteins to approximately 90% purity with relatively high yields (3 mg per liter of culture). Consistent with the result in mammalian cells, SOD1 (G93A) was more insoluble than wild-type SOD1 in E. coli, indicating that research on the aggregate formation of SOD1 may be possible using this pGEX expression system in E. coli. We investigated the HtrA2 serine protease activity on SOD1 to assess the relationship between two proteins. Not only wild-type SOD1 but also ALS-associated mutant SOD1 (G93A) were cleaved by HtrA2, resulting in the production of the 19 kDa and 21 kDa fragments that were specific for anti-SOD1 antibody. Using protein gel electrophoresis and immunoblot assay, we compared the relative molecular masses of thrombin-cleaved GST-SOD1 and HtrA2-cleaved SOD1 fragments and can predict that the HtrA2-cleavage sites within SOD1 are the peptide bonds between leucine 9-lysine 10 (L9-K10) and glutamine 23-lysine 24 (Q23-K24). Our study indicates that SOD1 is one of the substrate for HtrA2, suggesting that both HtrA2 and SOD1 may be important for modulating the HtrA2-SOD1-mediated apopotic cell death that is associated with the pathogenesis of neurodegenerative disorder.

Protective Effect of White-Skinned Sweet Potato (Ipomoea batatas L.) from Indonesia on Streptozotocin-Induced Oxidative Stress in Rats (흰 쥐에서 streptozotocin으로 유발된 산화적 스트레스에 대한 인도네시아산 white-skinned sweet potato (WSSP, Ipomoea batatas L.)의 보호효과)

  • Bachri, Moch. Saiful;Jang, Hye-Won;Choi, Jong-Won;Park, Jong-Ok
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1569-1576
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    • 2010
  • Sweet potato (Ipomoea batatas L.) is widely used in Indonesia and other countries as a traditional medicine for the treatment of diabetes mellitus (DM). The MeOH extract of white skinned sweet potatoes (WSSP) was administered orally in doses of 100 and 200 mg/kg body weight in streptozotocin (STZ)-induced diabetic rats. Experimental diabetes was induced by a single dose of STZ (45 mg/kg, i.p.) injection. Oxidative stress was measured by tissue lipid peroxide (LPO) levels, serum aspartate transaminase (AST), alanine transaminase (ALT), total triglyceride (TG), total cholesterol (TC) and by antioxidative enzymatic activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione S-transferase in the liver. An increase in blood glucose, LPO level, AST, ALT, TG and TC levels was observed in the STZ-induced diabetic rats. Administration of MeOH extract of WSSP at a dose of 200 mg/kg for two weeks caused a significant reduction in blood glucose, LPO levels, AST, ALT, TG and TC levels in the STZ-induced diabetic rats. Furthermore, oral administration of MeOH extract showed significant improvement in the activities of antioxidant enzymes (SOD, GPx, and CAT) compared to STZ-induced diabetic rats. In conclusion, the obtained results clearly indicate the role of oxidative stress in the induction of diabetes, and that the protective effects of MeOH extracts of WSSP could be used to benefit diabetic patients.

The Effects of Chungpyesagan-tang on the Recovery of Liver Function in Rat Injured by $CCl_4$ (청폐사간탕(淸肺瀉肝湯)이 $CCl_4$로 유발(誘發)된 휜쥐의 간손상(肝損傷)에 마치는 영향(影響))

  • Shin, Mee-Ran;Heo, Woon-Yeong;Kim, DaI-Rae;Jeon, Jong-Weon;Kim, Jeong-Yeol
    • Journal of Sasang Constitutional Medicine
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    • v.17 no.1
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    • pp.130-141
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    • 2005
  • 1. Objectives The present study was carried out to investigate the effects of Chungpyesagan-tang on the $CCI_4-induced$ Liver Damage in Rats. 2. Methods Sprague-Dawley rats were devided into 5 experimental groups : Normal, $NS+CCI_4(Solid$ extract of $CCI_4$ injection group after Normal Saline feed), $CP+CCI_4(Solid$ extract of $CCI_4$ injection group after Chungpyesagantang feed), $CCI_4+NS(Nomal$ Saline feed group after $CCI_4$ injection), $CCI_4+CP(Solid$ extract of Chungpyesagantang feed group after $CCI_4$ injection). Biochemical assays for serum enzyme activities such as AST, ALT, ALP, BUN, Creatinine, Uric Acid, Total Protein, Albumin, Total Cholesterol, Triglyceride, Glucose, and mRNA Revelation of Cytochrome p450 and activities such as LPO(Lipid Peroxidation), GSH(Glutathione), GST(Glutathione-S- Transferase), Glutathione Reductase, Glutathione Peroxidase, SOD(Superoxide Dismutase), Catalase, Hydroxyproline, and ${\beta}-Glucuronidase$ were performed. 3. Results 1) $CP+CCI_4$ showed significantly lower relation of Cytochrome p450 than $NS+CCI_4$. 2) As to LPO Hydroxyproline, $CCI_4+CP$ showed significantly lower activity than $CCI_4+NS$. 3) As to GSH GST Glutathione Peroxidase Catalase, $CP+CCI_4$ showed higher activity than $NS+CCI_4$, $CCI_4+CP$ showed significantly higher activity than $CCI_4+NS$. 4) As to Glutathione Reductase SOD, $CCI_4+CP$ showed significantly higher activity than $CCI_4+NS$. 5) As to ${\beta}-Glucuronidase$, $CP+CCI_4$ showed signficantly lower activity than $NS+CCI_4$. 4. Conclusions Chungpyesagantang has the recovering effects on the $CCI_4-induced$ Liver Damage.

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Role of Rab11 on Membrane Trafficking of Rat Vanilloid Receptor, TRPV1 (바닐로이드 수용체 TRPV1의 막수송과정에서의 Rab11의 역할)

  • Um, Ki-Bum;Lee, Soon-Youl
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.12 no.7
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    • pp.3096-3102
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    • 2011
  • Vanilloid receptor, TRPV1 (transient receptor potential vanilloid 1) is a non-selective cation channel that responds to a variety of pain-eliciting material including capsaicin, pH, heat. Although, membrane trafficking of TRPV1 was not much known so far, TRPV1 was reported to interact with FIP3 (family of Rab11 interacting protein 3). FIP3 was identified as one of Rab11 interacting proteins that is recently reported important in membrane trafficking of several channel proteins directly or indirectly. Therefore, in this study, we examined the role of Rab11 in the membrane trafficking of TRPV1 using cell biological and biochemical techniques. Rab11 was found really colocalized with TRPV1 based on the result of confocal microscopy. However, GST-pulldown assay, one of biochemical technique, found that Rab11 did not interact with TRPV1. Although Rab11 does not interact with TRPV1 directly, we hypothesized that Rab11 is indeed involved in the membrane trafficking of TRPV1. In order to examine further the role of Rab11 in the membrane trafficking of TRPV1, the expression of TRPV1 on the membrane was examined when the expression of Rab11 was decreased down to about 50% by siRNA technique and found decreased significantly. From this result, we can conclude that Rab11 is involved in the membrane trafficking of TRPV1 in a way of including FIP3.

The Expression and Functional Analysis of Recombinant Alcohol Dehydrogenase (재조합 alcohol dehydrogenase의 발현 및 기능분석)

  • Kong, Kwang-Hoon;Shim, Eun-Jung;Park, Hee-Joong;Kim, Eun-Ho;Cho, Sung-Hye;Park, Sung-Woo;Kim, Young-Mann
    • Analytical Science and Technology
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    • v.12 no.6
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    • pp.565-570
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    • 1999
  • The alcohol dehydrogenase (ADH) gene from Bacillus stearothermopilus was amplified by the polymerase chain reaction. The amplified DNA was inserted into the expression vector pGEX-KG, and expressed it as a fusion protein with glutathione S-transferase (GST) in E. coli. The recombinant ADH was produced by induction with 1 mM isopropyl-${\beta}$-D-thiogalactopyranoside at $37^{\circ}C$ and purified by glutathione affinity chromatography. The recombinant ADH exhibited high substrate specificity for ethanol. The activity of the recombinant ADH proceeded optimally at pH 9.0 and $70^{\circ}C$. The recombinant ADH was highly stable against high temperature. This thermostable alcohol dehydrogenase can be used for the enzymatic determination of alcohol and for the industrial production of alcohol.

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A Study on the Oxidative Damage Induced by UVB Irradiation to Mouse Skin (UVB 조사로 인한 마우스 피부조직의 산화적 손상)

  • Rhie Sung-Ja;Kim Young-Chul
    • Environmental Analysis Health and Toxicology
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    • v.21 no.2 s.53
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    • pp.165-172
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    • 2006
  • The backs with a hair cut of 6-week-old healthy ICR male mice were once exposed to a dose of $400mJ/cm^2$ UVB. An acute dermal inflammation was observed, and the inflamed skins were almost completely cured after 6 days of the exposure. At 24 hours after exposure, the epidermal keratinocytes showed a cell-membrane damage with the destruction of intercellular junctions, agglutination of tonofilaments within the cytoplasm and nucleus damage. The activity of XO showed a significant increase (p<0.05) in up to 144 hours. The activities of CAT and SOD showed a significant decrease (p<0.05) in up to 96 hours, but they were not significantly different from the normal value at 144 hours. The GST activity was significantly decreased (p<0.01) in up to 96 hours, not so at 24 hours. However, that was not significantly different from the normal value at 144 hours. There was a significant decrease (p<0.01) in the contents of TBARS at 48 and 96 hours, without any significant difference at 144 hours. While the content of GSH was significantly lower (p<0.05) at 24 hours, that was not significantly different thereafter up to 144 hours from the normal value. Therefore, it is assumed that skin damage with a dose of $400mJ/cm^2$ UVB irradiation might be caused by the oxidative stress which was resulted from the unbalance of oxygen fret radical generating and scavenging enzymes.

Isoegomaketone Upregulates Heme Oxygenase-1 in RAW264.7 Cells via ROS/p38 MAPK/Nrf2 Pathway

  • Jin, Chang Hyun;So, Yang Kang;Han, Sung Nim;Kim, Jin-Baek
    • Biomolecules & Therapeutics
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    • v.24 no.5
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    • pp.510-516
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    • 2016
  • Isoegomaketone (IK) was isolated from Perilla frutescens, which has been widely used as a food in Asian cuisine, and evaluated for its biological activity. We have already confirmed that IK induced the HO-1 expression via Nrf2 activation in RAW264.7 cells. In this study, we investigated the effect of IK on the mechanism of HO-1 expression. IK upregulated HO-1 mRNA and protein expression in a dose dependent manner. The level of HO-1 mRNA peaked at 4 h after $15{\mu}M$ IK treatment. To investigate the mechanisms of HO-1 expression modulation by IK, we used pharmacological inhibitors for the protein kinase C (PKC) family, PI3K, and p38 MAPK. IK-induced HO-1 mRNA expression was only suppressed by SB203580, a specific inhibitor of p38 MAPK. ROS scavengers (N-acetyl-L-cysteine, NAC, and glutathione, GSH) also blocked the IK-induced ROS production and HO-1 expression. Furthermore, both NAC and SB203580 suppressed the IK-induced Nrf2 activation. In addition, ROS scavengers suppressed other oxidative enzymes such as catalase (CAT), glutathione S-transferase (GST), and NADH quinone oxidoreductase (NQO-1) in IK-treated RAW264.7 cells. Taken together, it can be concluded that IK induced the HO-1 expression through the ROS/p38 MAPK/Nrf2 pathway in RAW264.7 cells.

Genetic Diversity and Phylogenetic Relationship of Genus Phyllostachys by RAPD Markers (RAPD분자 마커를 이용한 왕대속 대나무의 유전적 다양성 및 계통 관계)

  • Lee, Song-Jin;Huh, Man-Kyu;Shin, Hyun-Cheol;Huh, Hong-Wook
    • Journal of Life Science
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    • v.20 no.6
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    • pp.819-824
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    • 2010
  • Genus Phyllostachys is a long-lived woody species primarily distributed throughout South East Asia. Many species of this genus has been regarded as medically and ecologically important in the world. We evaluated representative samples of the four taxa with RAPD to estimate genetic relationships within the genus Phyllostachys. The percentages of polymorphic loci were 8.9-33.3% at the species level. P. bambusoides was found to show lower genetic diversity (H=0.018) than other species. Total genetic diversity ($H_T$) was 0.315, genetic diversity within populations ($H_S$) was 0.043, the proportion of total genetic diversity partitioned among populations ($G_{ST}$) was 0.659 and the gene flow (Nm) was 0.0263. As some Korean populations were isolated and patchily distributed, they exhibited low levels of genetic diversity. The four taxa of the genus Phyllostachys analyzed were distinctly related to a monophyletic. P. nigra var. henonis. Stapf was found to be more closely related to P. pubescens than to P. nigra. P. bambusoides was quite distinct from the remaining species.