• Title/Summary/Keyword: Glutathione-S-transferase M1

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SPR-based Antibody-Antigen Interaction for Real Time Analysis of Carbamate Pesticide Residues

  • Yang, Gil-Mo;Kang, Suk-Won
    • Food Science and Biotechnology
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    • v.17 no.1
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    • pp.15-19
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    • 2008
  • This research was conducted to develop a quick and sensitive method of detecting carbamate residues using the immobilization of antibody-antigen interactions with surface plasmon resonance (SPR). We have used commercialized surface plasmon resonance equipment (Biacore 3000). The antibody used for the immunoassay was specific for glutathione-s-transferase (GST) and the antigens included several carbamate pesticides (carbofuran, carbaryl, and benfuracarb). When antigens were applied to the protein GST, the detection limit was 2 ng/mL of carbamate pesticide. The fabricated protein GST maintained its activity for over 200 measurements. Thus we determined that the SPR biosensors could detect the specific reversible binding of a reactant in solution to a binding partner immobilized on the surface of the sensor and allow real-time detection and monitoring.

Detection of Multi-class Pesticide Residues Using Surface Plasmon Resonance Based on Polyclonal Antibody

  • Yang, Gil-Mo;Kang, Suk-Won
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.547-552
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    • 2008
  • The detection of carbamate (carbofuran, carbaryl, benfracarb, thiodicarb, and methomil) and organophosphate (diazinon, cadusafos, ethoprofos, parathion-methyl, and chlorpyrifos) pesticide residues with very low detection limits was carried out using surface plasmon resonance (SPR) based equipment. The capacity to develop a portable SPR biosensor for food safety was also investigated. The applied ligand for the immunoassays was polyclonal goat anti-rabbit immunoglobulin (IgG) peroxidase conjugate. Concentration tests using direct binding assays showed the possibility of quantitative analysis. For ligand fishing to find a proper antibody to respond to each pesticide, acetylcholinesterase (AChE), and glutathione-S-transferase (GST) were tested. The reproducibility and precision of SPR measurements were evaluated. With this approach, the limit of detection for pesticide residues was 1 ng/mL and analysis took less than 11 min. Thus, it was demonstrated that detecting multi-class pesticide residues using SPR and IgG antibodies provides enough sensitivity and speed for use in portable SPR biosensors.

Effects of Dietary Zinc Supplements on the Antioxidant Indicators and the Expression of Zinc Transport Genes in Korean Native Chicks (한국 재래닭에서 아연 보충급여가 항산화 지표 및 아연 운반 유전자 발현에 미치는 영향)

  • Jeon, Dong-Gyung;Kim, Min-Jeong;Yoon, Il-Gyu;Ahn, Ho-Sung;Sohn, Sea-Hwan;Jang, In-Surk
    • Korean Journal of Poultry Science
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    • v.46 no.3
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    • pp.161-171
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    • 2019
  • Four-week-old male Korean native chicks (KNC) were assigned to 3 groups with 6 replicates (8 birds/replicate) in each group: a basal diet (CON, 100 ppm of Zn), basal diet fortified with 50 ppm of Zn with zinc oxide (ZnO), or basal diet fortified with 50 ppm of Zn with Zn-methionine (ZnM). Immediately after a 4-week-feeding trial, 6 birds per group were used to evaluate the effects of zinc supplements on antioxidant indicators and the mRNA expression of zinc transport genes. The nitrogen components, lipid peroxidation, and total antioxidant status in blood were not influenced by Zn fortified diets. However, the ZnM group showed a significant (P<0.05) increase in uric acid levels than those in the ZnO group. In the small intestine, superoxide dismutase (SOD) and glutathione peroxidase (GPX) activities, and malondialdehyde (MDA) level were unaffected by zinc supplements. The activity of glutathione S-transferase (GST) was significantly (P<0.05) enhanced by Zn-methionine supplementation. In the liver, the activity of GST was significantly (P<0.05) increased by Zn-methionine supplement without affecting SOD, GPX, and MDA levels. With respect to the mRNA expression of zinc transport genes, the ZnM group displayed a strong tendency for increases in intestinal ZnT-1 (P=0.09) and ZnT-5 (P=0.06) levels, compared to those in the CON group. Moreover, the ZnM group showed a tendency (P=0.10) for up-regulation of hepatic metallothionein mRNA as compared with the CON group. In conclusion, the Zn-fortified diet with 50 ppm of Zn-methionine helped to improve GST activity and Zn transport gene expression in the small intestine or liver of KNC.

Eliminatory Effect of Mixture including Hot Water Extract of Dendropanax morbifera Lev. on Alcohol-induced Blood Alcohol Concentration and Hangover in Rat (흰쥐에서 황칠나무 열수 추출물을 포함한 혼합물의 혈중 알코올 농도와 숙취 해소 효과)

  • Na, Ju-ryun;Kim, Eun;Park, Soyi;Lee, Kihoon;Jeong, Eui-seon;Kim, Jinseok;Kim, Youngjae;Kim, Sunoh
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.267-276
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    • 2018
  • This study was performed to investigate the ameliorating effect of a hangover beverage mixture (SBJ) that contains Dendropanax morbifera Lev. and several medicinal plant extracts, on hepatoprotection and alcohol-metabolizing enzymes in alcohol-induced hangover in both in vitro and in vivo models. In human hepatoma cell line, HepG2, 300 mM of ethanol-induced hepatotoxicity was significantly improved by pretreatment of SBJ by dose-dependent manner. In the in vivo study, administration of alcohol to rats raised to the concentration of blood alcohol and lactate dehydrogenase (LDH). Blood alcohol and LDH levels in SBJ-treated rats significantly decreased at 0.5 h and 8 h after acute ethanol administration (40%, 4.6 g/kg body weight) as compared to alcohol-treated rats. Hepatic alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activity were significantly higher in SBJ-treated rats than in alcohol-treated rats. SBJ supplementation reduced formation of malondialdehyde (MDA), and inhibited reductions of hepatic superoxide dismutase (SOD), hepatic glutathione (GSH), glutathione-S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx) levels, compared with rats administered alcohol. Plasma catalase (CAT), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels showed unaltered resulted in all experimental groups compared with the control group. These results suggest that SBJ exhibit hepatoprotective properties by enhancing ADH, ALDH activity and stimulating the antioxidant defense system in alcohol-induced hangover.

Distributions of the GSTM1 and GSTT1 Null Genotypes Worldwide are Characterized by Latitudinal Clines

  • Saitou, Marie;Ishida, Takafumi
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.1
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    • pp.355-361
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    • 2015
  • Background: Deletion types of genetic variants of glutathione S-transferase (GST) M1 and T1, the GSTM1 null and GSTT1 null which are risk factors for certain cancers, have been ubiquitously found in human populations but their worldwide distribution pattern is unclear. Materials and Methods: To perform a meta-analysis, a systematic search for the literature on GSTM1 and GSTT1 null genotypes was done to identify 63 reports for 81 human populations. Relationships between the GSTM1 and GSTT1 null genotype frequencies and the absolute latitude of 81 populations were tested by Spearman's rank correlation coefficient. Results: A significant positive correlation was detected between the GSTM1 null genotype frequency and the absolute latitude (r=0.28, p-value <0.05), whereas the GSTT1 null genotype frequency and absolute latitude showed a significant negative correlation (r= -0.41 p-value <0.01). There was no correlation between the frequencies of GSTM1 and GSTT1 null genotype in each population (r= -0.029, p-value=0.80). Conclusions: Latitudinal clines of the distribution of the GSTM1 and GSTT1 null genotypes may be attributed to the result of gene-environmental adaptation. No functional compensation between GSTM1 and GSTT1 was suggested by the lack of correlation between the null frequencies for GSTM1 and GSTT1.

Induction of Oxidative Stress by Hexavalent Chromium in Human Bronchial Epithelial Cells (BEAS-2B) (배양 기관지 상피세포(BEAS-2B cells)에서 6가 크롬에 의한 산화적 스트레스)

  • Park, Eun-Jung;Kang, Mi-Sun;Kim, Dae-Seon;Park, Kwang-Sik
    • Environmental Analysis Health and Toxicology
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    • v.21 no.4 s.55
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    • pp.357-363
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    • 2006
  • Chromium compounds are widely used in diverse industries including pigment manufacturing, painting, metal plating and leather tanning. With the wide uses of chromium, various adverse effects of the compounds on the environment and human health have been reported. Among them, hexavalent chromium [Cr (VI)], which is a carcinogenic heavy metal, has been widely studies. Epidemiological investigations have shown that respiratory cancers had been found in workers who had been occupationally exposed to Cr (VI). In this study, cell toxicity and induction of reactive oxygen species (ROS) by Cr (VI) (1, 2, 4, $8{\mu}M$) in cultured human bronchial epithelial cells were investigated. Exposure of the cells to Cr (VI) led to cell death, ROS increase, and cytosolic caspase-3 activation. The ROS increase was related with the decreased level of GSH. Chromatin condensation and fragmentation were occurred by Cr (VI) when evaluated by DAPI staining or agarose gel electrophoresis of the extracted DNA. Expression of ROS related genes including glutathione S-transferase, heme oxygenase-1, metallothionein were significantly induced in Cr (VI) treated cells. This result suggests the toxicity in cultured cells by Cr (VI) was expressed through the apoptotic process with ROS induction.

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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IMMUNOREGULATORY EFFECTS OF A MONOCLONAL ANTIBODY TO HUMAN 4-1 BB MOLECULE ON ALLOANTIGEN-MEDIATED IMMUNE RESPONSES.

  • Kim, Joong-Gon;Lee, Soo-Hyun;Lee, Jae-Woo;B.S. Kwon;Kang, Chang-Yuil
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.82-82
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    • 1995
  • 4-lBB molecule is expressed on the surface of activated CD4$\^$+/ and CD8$\^$+/ T cells. We generated a panel of anti-4-1 B5 murine mAbs using a fusion protein consisting of the extracellular domain of human 4-1 BB fused to Glutathione S-transferase. The binding activity against cell surface 4-1 BB molecule was assessed by flow cytometry analysis. These studies showed that several anti-4-1 BB mAbs bound to 10-30% of CD4$\^$+/ and CD8$\^$+/T cells in PHA or Con A stimulated PBLs, although these mAbs interacted with only, l-2% of CD4$\^$+/ and CD8$\^$+/ T cells in normal PBLs, indicating the specificity of mAbs to the 4-l BB molecule on activated CD4$\^$+/ and CD8$\^$+/ T cells. Next, we examined the effect of an anti-4-l BB mAb (4B4-1-1) on allogeneic mixed lymphocyte reactions (MLRs). The data indicated that the antibody significantly inhibited the proliferative response at higher concentrations. When tested with several T cell mitogens, the antibody had no stimulatory or inhibitory effects on the mitogen-mediated T cell proliferation. These data suggest that 4-1 BB molecule may play a role in the regulation of antigen-mediated immune response.

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Molecular Parameters for Assessing Marine Biotoxicity: Gene Expressions of Rockfish (Sebastes schlegeli) Exposed to Polycyclic Aromatic Hydrocarbons

  • Woo, Seon-Ock;Yum, Seung-Shic;Park, Hong-Seog;Jung, Jee-Hyun;Lee, Suk-Chan;Kim, So-Jung;Lee, Taek-Kyun
    • Molecular & Cellular Toxicology
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    • v.3 no.4
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    • pp.267-272
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    • 2007
  • Environmental and anthropogenic changes affect the health and stability of marine ecosystem. In this study we aimed to identify molecular biomarkers for ecotoxicological pollutants risk assessment in the rockfish (Sebastes schlegeli). We designed primers based on conserved sequences by multiple alignments of target genes from related species, and cloned the partial cDNAs of cytochrome P450 (CYP1A1), glutathione S-transferase (GST), metallothionein (MT), superoxide dismutase (SOD), ubiquitin (UB), vitellogenin (VTG) and $\beta$-actin by reverse transcription polymerase chain reaction (RT-PCR) from S. schlegeli. Northern blot results indicated that these six genes expressions were significantly induced by benzo[a]pyrene (BaP, 1 ${\mu}M$) and that the level of each of their transcripts increased in BaP-exposed rockfish in a time-dependent manner. This study suggests that transcriptional changes in these six genes may be used for monitoring environmental exposure to polycyclic aromatic hydrocarbons (PAHs).

No Association Between the GSTM1 Null Genotype and Risk of Renal Cell Carcinoma: A Meta-analysis

  • Liu, Rui;Wang, Xiao-Hua;Liu, Li;Zhou, Qiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3109-3112
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    • 2012
  • Background: Many studies have focused on possible associations between the glutathione S-transferase M 1 (GSTM1) null genotype and risk of renal cell carcinoma (RCC), but the impact remains unclear owing to obvious inconsistencies among the findings. The present study aimed to quantify the strength of any association in a meta-analysis. Methods: We searched the PubMed, Embase and CBM databases for studies concerning the association between the GSTM1 null genotype and risk of RCC. We estimated the summary odds ratio (OR) with its 95% confidence intervals (95% CI) to assess the association. Results: The meta-analysis showed the GSTM1 null genotype was not associated with risk of RCC overall (OR = 1.04, 95% CI 0.92-1.18, P = 0.501). For Caucasians, the GSTM1 null genotype was also not associated with risk of RCC (OR=1.02, 95% CI 0.90-1.16, P = 0.761). The cumulative meta-analyses showed a trend of no obvious association between GSTM1 null genotype and risk of RCC as information accumulated. Sensitivity analyses by omitting those studies also did not materially alter the overall combined ORs. No evidence of publication bias was observed. Conclusion: Meta-analyses of available data show that the GSTM1 null genotype is not significantly associated with risk of renal cell carcinoma.