• Title/Summary/Keyword: GST-P

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Effects of Fluoride Levels on Lipid Peroxidation and Antioxidant Systems of Growing/Finishing Pigs

  • Tao, X.;Xu, Z.R.;Han, X.Y.;Wang, Y.Z.;Zhou, L.H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권4호
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    • pp.552-556
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    • 2005
  • Malondialdehyde (MDA) and total antioxidant capacity (T-AOC) levels, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), glutathione transferase (GST) and xanthine oxidase (XOD) activities were analyzed in serum, livers and kidneys of pigs treated with graded doses of fluoride (as NaF). Ninety-six Duroc-Landrace-Yorkshire crossbred growing pigs (48 barrows and 48 gilts, respectively), with similar initial weight 24.14${\pm}$1.12kg, were randomly assigned to four different treatments. These treatments containing the following added F: basal control; 50 mg/kg F; 100 mg/kg F and 150 mg/kg F were randomly assigned to four pens (three barrows and three gilts) each in a completely randomized design. The results showed pigs treated with 150 mg/kg F significantly decreased average daily gain (ADG) (p<0.05) and increased feed/gain ratio (F/G) (p<0.05) compared to the controls. In the groups treated with fluoride, the contents of MDA increased, T-AOC levels and the activities of SOD, GSH-PX, CAT, GST and XOD decreased, and most of which altered significantly (p<0.05). The study therefore indicated the mechanism of excess fluoride on the impairment of soft tissues involved in lipid peroxidation and decreased the activities of some enzymes associated with free radical metabolism.

지방산으로 유발된 비알코올지방간질환 실험모델에 미치는 생간탕가미방(生肝湯加味方)의 효과 (Effects of Saenggantanggami-bang on Nonalcoholic Fatty Liver Disease Model Induced by Fatty Acids)

  • 유재연;이장훈
    • 대한한방내과학회지
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    • 제31권2호
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    • pp.331-345
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    • 2010
  • Objectives : This study was designed to investigate the effects of Saenggantanggami-bang (SG) on nonalcoholic fatty liver disease. Methods : HepG2 cells were used in an in vitro model. HepG2 cells were divided into three groups. The Normal group was incubated with no fatty acid. The Control group was incubated with 1mM palmitic acid to introduce fat overloading. The PA-SG group was incubated with 1mM palmitic acid and various concentrations of Saenggantanggami-bang (SG). Cell viability and cytotoxicity were analyzed by MTT assay and LDH assay. Intracellular triglyceride (TG) levels, reactive oxygen species (ROS) levels, ATP amount, and GST activity were measured. Cell death pattern and protective effect of SG on cell death were studied by DNA fragmentation and caspase-3 intensity (western blot). Results : Compared with the Control group, cell viability of the PA-SG group significantly increased (P<0.01), cytotoxicity of the PA-SG group decreased (P<0.01), and intracellular TG levels and ROS levels of the PA-SG group decreased (P<0.05). In DNA fragmentation assay, necrotic pattern was observed and DNA fragment decreased in the PA-SG group. In western blot, apoptotic pattern was observed, caspase-3 intensity of the PA-SG group was reduced significantly, but there were no significant differences in intracellular ATP amount and GST activity between the control group and the PA-SG group. Conclusion : The results suggest that Saenggantanggami-bang can be a potential candidate for the clinical treatment of nonalcoholic fatty liver disease.

Comparative Investigation of Glutathione S-Transferases, Glyoxalase-I and Alliinase Activities in Different Vegetable Crops

  • Hossain, Md Daud;Rohman, Md Motiar;Fujita, Masayuki
    • Journal of Crop Science and Biotechnology
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    • 제10권1호
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    • pp.19-26
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    • 2007
  • Glutathione S-transferases(GSTs, EC 2.5.1.18), glyoxalase-I(EC 4.4.1.5) and alliin lyase(alliinase, EC 4.4.1.4) are important enzyme systems in plant bodies. The first two are mainly detoxifying enzymes that utilize glutathione(GSH) in the defense mechanism, and the last one is mainly involved in secondary metabolism and relevant to sulfur compounds derived from GSH. The activities of the three enzymes have been investigated in soluble extracts of vegetable crops, including pumpkin, cabbage, broccoli, radish, carrot, potato, sweet potato, mungbean, and onion. GST activities were detected in all of the vegetables, and the extract of onion bulb exhibited the highest specific activity(648 nmol/min/mgP). The putative GSTs of most of the vegetables were found to be induced by ethanol. The activities of GSTs in onion bulb were found to be markedly inhibited by S-hexyl glutathione and were also inhibited by S-butyl glutathione and S-propyl glutathione. The anti-CmGSTF1 antiserum recognized a thick band for putative onion GST. The estimated glyoxalase-I activity level was also high in onion bulb(4540 nmol/min/mgP), indicating that the thick band detected by Western blot analysis might result from partial recognition of glyoxalase-I by the antiserum. The specific activities for glyoxalase-I were moderate in radish and carrot, and the extracts of other vegetables had rather low levels of activities. The extract of onion also showed the highest specific activity level for alliinase(2069nmol pyruvate/mgP). The extracts of other vegetables also had alliinase activities, although the estimated values were much lower than that of onion.

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Production and Purification of Single Chain Human Insulin Precursors with Various Fusion Peptides

  • Cho, Chung-Woo;Park, Sun-Ho;Nam, Doo-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제6권2호
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    • pp.144-149
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    • 2001
  • For the production and purification of a single chain human insulin precursor, four types of fusion peptides $\beta$-galactosidase (LacZ), maltose binding protein (MBP), glutathione-S-transferase (GST), and (His)(sub)6-tagged sequence (HTS) were investigated. Recombinant E. coli harboring hybrid genes was cultivated at 37$\^{C}$ for 1h, and gene induction occurred when 0.2mM of isopropyl-D-thiogalactoside (IPTG) was added to the culture broth, except for E. coli BL21 (DE3) pLysS harboring a pET-BA cultivation with 1.0mM IPTG, followed by a longer than 4h batch fermentation respectively. DEAE-Sphacel and Sephadex G-200 gel filtration chromatography, amylose affinity chromatography, glutathione-sepharose 4B affinity chromatography, and a nickel chelating affinity chromatography system as a kind of immobilized metal ion affinity chromatography (IMAC) were all employed for the purification of a single chain human insulin precursor. The recovery yields of the HTS-fused, GST-fused, MBP-fused, and LacZ-fused single chain human insulin precursors resulted in 47%, 20%, 20%, and 18% as the total protein amounts respectively. These results show that a higher recovery yield of the finally purified recombinant peptides was achieved when affinity column chromatography was employed and when the fused peptide had a smaller molecular weight. In addition the pET expression system gave the highest productivity of a fused insulin precursor due to a two-step regulation of the gene expression, and the HTS-fused system provided the highest recovery of a fused insulin precursor based on a simple and specific separation using the IMAC technique.

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The Anti-hepatotoxic Effect of Ginseng in Rats: Meta-analysis

  • Kook, Se-Jeong;Han, Hye-Kyoung;Kim, Gun-Hee;Choi, Ki-Heon
    • Journal of Ginseng Research
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    • 제32권2호
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    • pp.161-170
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    • 2008
  • The aim of this meta-analysis was to systematically investigate the anti-hepatotoxic effect of ginseng in rats induced toxicity which damage to liver. Primary researches were gained on the ScienceDirect database, the DBpia, and the KISS, and the data about the effect factors in plasma and in enzyme were listed as many as possible. The effect factors were alanine transaminase (ALT), aspartate transaminase (AST), liver aminopyrine N-demethylase (AD), liver aniline hydroxylase (AH), liver 3,4-Methylenedioxyamphetamine (liver MDA), cytochrome P450 (P450), serum alkaline phosphatase (ALP), serum lactate dehydrogenase (LDH), cytochrome b5 (Cyto b5), glutathione reductase (GR), Liver glutathione S-transferase (GST), liver glutamyltransferase (GT), Liver (${\gamma}-GCS$), serum liver 3,4-Methylenedioxyamphetamine (serum MDA), serum sorbitol dehydrogenase (SDH), serum total protein (TP), serum ${\gamma}-glutamyltransferase$ (${\gamma}-GT$). To investigate the effect of ginseng, the mean difference (MD) between the group of rats induced by toxicity (RH) and the group of rats induced by toxicity with ginseng (RHG) were combined, and the significance of MDs were tested. The combined MDs were checked the biases caused by heterogeneity among studies and the publication biases, and adjusted by using random effect model and trim and fill method, respectively. The effect about ALT, AST, ALP, LDH, SDH, TP and ${\gamma}-GT$ in plasma factors were significant, and about AD, liver MDA, P450, Cyto b5, GR, GST, GT and ${\gamma}-GCS$ in enzyme factors were significant. The treatment with ginseng supplementation was significantly effected on plasma and enzyme factors of damaged-rats.

Cloning and overexpression of lysozyme from Spodoptera litura in prokaryotic system

  • Kim, Jong-Wan;Park, Soon-Ik;Yoe, Jee-Hyun;Yoe, Sung-Moon
    • Animal cells and systems
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    • 제15권1호
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    • pp.29-36
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    • 2011
  • Insect lysozymes are basic, cationic proteins synthesized in fat body and hemocytes in response to bacterial infections and depolymerize the bacterial cell wall. The c-type lysozyme of the insect Spodoptera litura (SLLyz) is a single polypeptide chain of 121 residues with four disulfide bridges and 17 rare codons and is approximately 15 kDa. The full-length SLLyz cDNA is 1039 bp long with a poly(A) tail, and contains an open reading frame of 426 bp long (including the termination codon), flanked by a 54 bp long 5' UTR and a 559 bp long 3' UTR. As a host for the production of high-level recombinant proteins, E. coli is used most commonly because of its low cost and short generation time. However, the soluble expression of heterologous proteins in E. coli is not trivial, especially for disulfide-bonded proteins. In order to prevent inclusion body formation, GST was selected as a fusion partner to enhance the solubility of recombinant protein, and fused to the amplified products encoding mature SLLyz. The expression vector pGEX-4T-1/rSLLyz was then transformed into E. coli BL21(DE3)pLysS for soluble expression of rSLLyz, and the soluble fusion protein was purified successfully. Inhibition zone assay demonstrated that rSLLyz showed antibacterial activity against B. megaterium. These results demonstrate that the GST fusion expression system in E. coli described in this study is efficient and inexpensive in producing a disulfide-bonded rSLLyz in soluble, active form, and suggest that the insect lysozyme is an interesting system for future structural and functional studies.

Quizalofop-Ethyl이 콩과 옥수수의 Glutathione-S-Transferases와 Carboxylesterase의 활성에 미치는 영향 (Effect of Quizalofop-Ethyl on Glutathione-S-Transferases and Carboxylesterase Activity of Soybean and Corn Plants)

  • 김희권;김명석;박인진;서용택
    • 한국환경농학회지
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    • 제16권4호
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    • pp.365-372
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    • 1997
  • 콩과 옥수수에서 glutatione-S-transferases와 carboxylesterase의 생화학적 특성과 quizalofopethyl 처리에 따른 식물체내 효소의 활성 변화를 조사한 결과는 다음과 같다. GSTS의 최적 pH는 8.2이고, Km값은 콩 $6.7{\times}10^{-3}M$, 옥수수 $2.5{\times}10^{-3}M$이었으며, Vmax는 콩 50nmole/mg/min, 옥수수 20nmole/mg/min이었다. Carboxylesterase의 최적 pH는 6.2${\sim}$7.0 범위이고, Km값은 콩 $4.2{\times}10^{-4}M$, 옥수수 $2.5{\times}10^{-4}M$이었으며, Vmax는 콩 33 nmole/mg/min, 옥수수 10nmole/mg/min이었다. 콩과 옥수수에 살포된 quizalofop-ethyl은 콩과 옥수수의 GSTs와 carboxy-lesterase의 활성을 감소시켰고, quizalofop-ethyl 80ppm을 처리했을때 콩은 10일이 경과후 회복되었지만, 옥수수는 3일이 경과후 고사하였다.

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Fatty acid compositions, free radical scavenging activities, and antioxidative enzyme activities of high-preference and low-preference beef cuts of Hanwoo (Bos taurus coreanae) cows

  • Moon, Sang-Ho;Kim, Eun-Kyung;Jang, Se Young;Tang, Yujiao;Seong, Hye-Jin;Yun, Yeong Sik;Chung, Sanguk;Oh, Mirae
    • Asian-Australasian Journal of Animal Sciences
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    • 제31권12호
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    • pp.1974-1979
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    • 2018
  • Objective: This study compared fatty acid compositions and antioxidant activities of high-preference cuts (loin, tenderloin, and rib) and low-preference cuts (brisket, topside, and shank) of Hanwoo (Bos taurus coreanae) cows to obtain potentially useful information for promoting the consumption of various low-preference cuts. Methods: Individual 500 g samples of fresh beef were collected from each of the six cuts from 10 Hanwoo cows (quality grade 1) and immediately freeze-dried. The dried samples were evaluated for fatty acid composition, free radical scavenging activities (hydroxyl, alkyl, and 2, 2'-diphenyl-1-picrylhydrazyl [DPPH] radical), and antioxidative enzyme activities (glutathione peroxidase [GPx], glutathione-S-transferase [GST], and superoxide dismutase [SOD]). Results: The percentages of total polyunsaturated fatty acids were significantly higher in low-preference cuts than in high-preference cuts (p<0.05). Hydroxyl, alkyl, and DPPH radical scavenging activities were significantly higher in low-preference cuts than in high-preference cuts (p<0.05). In addition, the activities of antioxidant enzymes, such as GPx, GST, and SOD, were significantly higher in low-preference cuts compared with high-preference cuts (p<0.05). Conclusion: These results may influence consumers to include more low-preference cuts in their selections based on the nutritional facts, which could help to balance the beef market in South Korea.

사염화탄소에 의한 간손상에 미치는 현호색의 효과 및 그 기전 (Mechanism and Effect of Corydalis ternata on the $CCl_4$-Induced$ Hepatotoxicity)

  • 서인옥;정춘식;정기화
    • 한국식품위생안전성학회지
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    • 제15권3호
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    • pp.226-234
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    • 2000
  • 본 연구에서는 현호색의 methanol추출물에 대하여 사염화탄소에 의한 간 손상 보호효과를 확인한 후 이를 hexane, chloroform, butanol 및 물로 계통분획하여 사염화탄소에 의한 간 손상 보호효과를 검색하였다. 이 결과 현호색 물분획물이 유의성 있는 간 보호효과를 보임으로써 간손상 보호효과를 확인하고 그 기전을 밝히고자 하였다. 그 결과 혈액중의 ALT활성 및 cholesterol함량은 사염화탄소 단독투여군에 비하여 감소하였으며 간조직의 TG와 지질과산화물 함량은 사염화탄소 단독투여군에 비하여 감소하여 사염화탄소에 의한 지방의 축적이나 세포막의 손상은 억제된 것으로 보인다. 기전연구를 위하여 측정한 CYP함량과 calcium함량은 사염화탄소 단독투여군에 비하여 감소하였으며 GST활성도는 사염화탄소 단독투여군에 비하여 증가하였다. 항산화 효소의 유도로 인하여 증가할 것으로 기대한 SOD, GPX, GST활성도는 모든 처치군에서 감소하는 경향을 보여 간손상 보호 효과는 free radical scavenging effect보다는 CYP함량의 감소와 CCB로서의 작용 및 GST활성도의 증가에 기인한 것으로 생각된다.

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살충제 Carbofuran과 Phenobarbital Sodium 및 3-Methylcholanthrene이 쥐의 효소활성에 미치는 영향 (Effect of Insecticide Carbofuran and Phenobarbital Sodium and 3-Methylcholanthrene on Activity of Enzyme in Rat)

  • 임요섭;한성수
    • 농약과학회지
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    • 제3권3호
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    • pp.27-36
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    • 1999
  • 쥐에 있어서 carbamate계 살충제 carbofuran의 독성에 미치는 phenobarbital sodium(PB) 또는 3-methylcholanthrene(3-MC)의 영향과 작용기작을 효소적 측면에서 구명할 목적으로 이들을 단독 또는 조합으로 경구투여 하여 in vivo 효소활성을 조사하였다. Acetylcholinesterase(AChE)와 butyrylcholinesterase(BuCheE)의 효소활성은 carbofuran 3.8 mg/kg을 투여하였을 때 48시간까지 $20{\sim}70%$ 범위의 저해를 보였고, carbofuran과 PB 또는 3-MC를 조합투여하였을 때 효소활성은 초기에 감소하다가 24시간 후에는 대조구와 비슷한 수준을 나타냈다. Glutathione S-transferase(GST)의 경우 carbofuran만을 투여하였을 때 초기($0.5{\sim}6$ hr)에 $15{\sim}35%$의 저해를 보였으나, carbofuran과 PB 또는 3-MC의 조합투여시 초기에는 약간 저해를 보이다가 3시간 후에는 대조군과 유사한 효소활성을 보였고, 6시간 후에는 대조군에 비해 활성이 20%이상 증가하였다. UDP-glucuronosyltransferase(UDPGI) 및 cytochrome P-450 효소계의 효소활성은 carbofuran과 PB 또는 3-MC를 조합 투여하였을 때 투여 후 6시간까지는 carbofuran만의 투여에 비해 효소활성이 $2.6{\sim}2.8$배 이상 높았다. 이상의 결과에서 PB 및 3-MC의 투여가 이들 효소활성을 유도하므로써 carbofuran의 독성으로부터 쥐를 보호한 것으로 판단된다.

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