• Title/Summary/Keyword: GST-P

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Role of Household Exposure, Dietary Habits and Glutathione S-Transferases M1, T1 Polymorphisms in Susceptibility to Lung Cancer among Women in Mizoram India

  • Phukan, Rup Kumar;Saikia, Bhaskar Jyoti;Borah, Prasanta Kumar;Zomawia, Eric;Sekhon, Gaganpreet Singh;Mahanta, Jagadish
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.3253-3260
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    • 2014
  • Background: A case-control study was conducted to evaluate the effect of household exposure, dietary habits, smoking and Glutathione S-Transferases M1, T1 polymorphisms on lung cancer among women in Mizoram, India. Materials and Methods: We selected 230 newly diagnosed primary lung cases and 460 controls from women in Mizoram. Multivariate logistic regression analysis was performed to estimate adjusted odds ratio (OR). Results: Exposure of cooking oil fumes (p<0.003), wood as heating source for cooking (p=0.004), kitchen inside living room (p=0.001), improper ventilated house (p=0.003), roasting of soda in kitchen (p=0.001), current smokers of tobacco (p=0.043), intake of smoked fish (p=0.006), smoked meat (p=0.001), Soda (p<0.001) and GSTM1 null genotype (p=0.003) were significantly associated with increased risk of lung cancer among women in Mizoram. Significantly protective effect was observed for intake of bamboo shoots (p=<0.001) and egg (p<0.001). A clear increase in dose response gradient was observed for total cooking dish years. Risk for lung cancer tends to increase with collegial effect of indoor environmental sources (p=0.022). Significant correlation was also observed for interaction of GST polymorphisms with some of dietary habits. Conclusions: We confirmed the important role of exposure of cooking oil emission and wood smoke, intake of smoked meat, smoked fish and soda (an alkali preparation used as food additives in Mizoram) and tobacco consumption for increase risk of lung cancer among Women in Mizoram.

Effects of Dietary Seaweed on Blood Glucose, Lipid and Glutathione Enzymes in Streptozotocin-Induced Diabetic Rats (미역첨가 식이가 당뇨 유발쥐의 혈청지질과 혈당 및 항산화 효소에 미치는 영향)

  • 조영자;방미애
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.6
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    • pp.987-994
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    • 2004
  • The purpose of this study was to investigate the effect of dietary seaweed in diabetic rats treated with streptozotocin (STZ) for 7 weeks. The rats (Sprague-Dawley male rats, 180∼200 g) were divided into 4 groups : normal rats fed control diet (C), diabetic rats fed control diet (CD), normal rats fed seaweed diet (M), and diabetic rats fed seaweed diet (MD). Diabetes was induced by single injection of streptozotocin (60 mg/kg, i.p.). Urinary levels of calcium and uric acid, and blood levels of hemoglobin, total cholesterol and low density lipoprotein (LDL)-cholesterol were not significantly different among groups. But high density lipoprotein (HDL)- cholesterol of M and MD groups were higher than that of C and CD groups. Activity of hepatic microsomal G6Pase was significantly (p<0.05) lower in C and M groups than that of CD and MD groups. Hepatic glutathione S-transferase (GST) of M, CD and MD groups were significantly lower than C group (p<0.05), glutathione peroxidase (GPX) of C, M and MD groups were higher than CD group. In conclusion, dietary seaweed may improve blood lipid profiles and GSH-related enzymes in STZ-induced diabetic rats.

Development of a Kit for Diagnosing AtCYP78A7 Protein in Abiotic-tolerant Transgenic Rice Overexpressing AtCYP78A7 (AtCYP78A7 과발현 환경스트레스 내성 형질전환 벼의 단백질 진단 키트 개발)

  • Nam, Kyong-Hee;Park, Jung-Ho;Pack, In-Soon;Kim, Ho Bang;Kim, Chang-Gi
    • Journal of Life Science
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    • v.28 no.7
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    • pp.835-840
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    • 2018
  • Quantitative determination of the protein expression levels is one of the most important parts in assessment of the safety of foods derived from genetically modified (GM) crops. Overexpression of AtCYP78A7, a gene encoding cytochrome P450 protein, has been reported to improve tolerance to abiotic stress, such as drought and salt stress, in transgenic rice (Oryza sativa L.). In the present study, an enzyme-linked immunosorbent assay (ELISA) kit for diagnosing AtCYP78A7 protein including AtCYP78A7-specific monoclonal antibody was developed. GST-AtCYP78A7 recombinant protein was induced and purified by affinity column. Four monoclonal antibodies (mAb 6A7, mAb 4C2, mAb 11H6, and mAb 7E8) against recombinant protein were also produced and biotinylated with avidin-HRP. After pairing test using GST-AtCYP78A7 protein and lysate of rice samples, mAb 4C2 and mAb 7E8 were selected as a capture antibody and a detecting antibody, respectively, for ELISA kit. Product test using rice samples indicated that percentages of detected protein in total protein were greater than 0.1% in AtCYP78A7-overexpressing transgenic rice (Line 10B-5 and 18A-4), whereas those in negative control non-transgenic rice (Ilpum and Hwayoung) were less than 0.1%. The ELISA kit developed in this study can be useful for the rapid detection and safety assessment of transgenic rice overexpressing AtCYP78A7.

Effect of Trace Metal on Accumulation and Physiological Response of the Polychaete, Perinereis nuntia (미량금속 노출에 따른 갯지렁이(Perinereis nuntia)의 미량금속 축적 및 생리학적인 반응)

  • Won, Eun-Ji;Ra, Kong-Tae;Hong, Seong-Jin;Kim, Kyung-Tae;Lee, Jae-Seong;Shin, Kyung-Hoon
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.13 no.4
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    • pp.288-295
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    • 2010
  • Metal exposure experiments using polychaete (Perinereis nuntia) as a bio-indicator of trace metals contamination were conducted to evaluate the bioaccumulation and the biomarkers responses such as metallothionein-like protein (MTLPs) and glutathione S-transferase (GST) which was simultaneously exposed to Cadmium (Cd) and Copper (Cu). Cu and Cd concentrations in polychaete were enhanced with increasing exposure time and their concentrations of aqueous medium. Initial accumulation of Cd was higher than that of Cu. Our results showed that the bioaccumulation of Cu and Cd were prohibited, especially at higher Cu levels, suggesting the different cellular uptake mechanisms when Cu and Cd are co-exist. Net accumulation rate of Cu was declined with exposure time but it did not show any significant change for Cd. Although the highest MTLPs concentration was observed at 6 hr of exposure time, it did not show any significant change related to exposure times and metals concentrations. An increase of GST activity tended to increase as a function of exposure time and metals concentrations. And GST activities in P. nuntia have similar tendency with bioconcentration factors in high concentration of Cu (treatment group IV) at post 24 h of exposure. Our results provide new information of the bioaccumulation and biomarker responses to understand the effects of co-existing contaminants (Cu and Cd) using polychaete. Further studies are required to elucidate the bioaccumulation and biomarkers responses for various contaminants.

Antioxidant Effect of Chungkukjang Supplementation against Memory Impairment induced by Scopolamine in Mice (Scopolamine으로 유도된 기억 손상 마우스에서 청국장 식이의 항산화 효과)

  • Kong, Hyun-Joo;Lee, Kyung-Eun;Yang, Kyung-Mi
    • Journal of the East Asian Society of Dietary Life
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    • v.26 no.3
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    • pp.237-249
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    • 2016
  • In this study, the antioxidant effect of Chungkukjang supplementation against memory impairment and oxidative stress in scopolamine (2 mg/kg i.p)-injected mice was investigated. The experimental animals were divided into five groups and fed experimental diets for 12 weeks; normal diet group (C), scopolamine + normal diet group (S), scopolamine + 63.0% soybean Chungkukjang supplementation group (SS), scopolamine + 45.0% Yakkong Chungkukjang supplementation group (SY), and scopolamine + 50.0% black foods such as black rice, black sesame seeds, and sea tangle added Yakkong Chungkukjang group (SYB). For the results of food intake, body weight gain, and brain weights, levels of scopolamine-injected groups were lower than the levels of the control group. The reduced brain weight of the scopolamine-injected group (S) was regulated to control level by supplementation of three types Chungkukjang. In the oxidative stress indicator, nitric oxide and malondialdehyde levels in serum of scopolamine-injected mice were higher than those of other groups. However, supplementation of soybeans, Yakkong and black foods added Yakkong Chungkukjang was proven to regulate them. Antioxidant enzyme activities such as superoxide dismutase (SOD) and glutathione-S-transferase (GST) in serum showed no significant differences among the groups. The reduced levels of vitamin A and vitamin E in serum and brain tissue of scopolamine-injected mice were controlled by supplementation of three types of Chungkukjang. Total antioxidant capacity (TAC) of scopolamine-injected group was lower than those of other groups. However, TAC was significantly elevated by Chunggukjang supplementation. Therefore, antioxidative effects of soybeans, Yakkong, and black foods added Yakkong Chungkukjang supplementations against oxidative stress in scopolamine-injected in mice could expected.

Effect of Ramaria botrytis Methanol Extract on Antioxidant Enzyme Activities in $Benzo({\alpha})Pyrene-treated$ Mice (싸리버섯 메탄올 추출물이 벤조피렌을 투여한 마우스의 항산화 효소 활성에 미치는 영향)

  • Kim, Hyun-Jeong;Lee, Kap-Rang
    • Korean Journal of Food Science and Technology
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    • v.35 no.2
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    • pp.286-290
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    • 2003
  • Effects of Ramaria botrytis methanol extract on hepatotoxicity in $benzo({\alpha})pyrene(B({\alpha})P)-treated$ mice were investigated. R. botrytis methanol extract was intraperitioneally injected once a day for successive 5 days, followed by treatment with $B({\alpha})P$ on the fifth day. Antioxidant activities of R. botrytis methanol extract were examined by measuring the free radical-scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. In DPPH method, R. botrytis methanol extract showed strong antioxidative activies. The increased activities of superoxide dismutase, catalase, and glutathione peroxidase after $B({\alpha})P-treatment$ were decreased by treatment of R. botrytis methanol extract. Glutathione content and glutathione S-transferase activity depleted by $B({\alpha})P$ were significantly increased, but elevation of lipid peroxide content induced by $B({\alpha})P$ was decreased by R. botrytis methanol extract. These results suggest that R. botrytis methanol extract is believe to be a possible protective effect against $B(\alpha)P-induced$ hepatotoxicity in mice.

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).

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.

Expression of Porcine Acid-labile Subunit (pALS) of the 150-kilodalton Ternary Insulin-like Growth Factor Complex and Initial Characterization of Recombinant pALS Protein

  • Lee, Dong-Hee;Chun, Choa;Kim, Sang-Hoon;Lee, C.-Young
    • BMB Reports
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    • v.38 no.2
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    • pp.225-231
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    • 2005
  • Acid-labile subunit (ALS) is a component of the 150-kDa insulin-like growth factor-binding protein-3 (IGFBP-3) complex, which, by sequestering the majority of IGFs-I and -II and thereby prolonging the half-life of them in plasma, serves as a circulating reservoir of IGFs in mammalian species. A pGEX-2T plasmid and a baculovirus expression constructs harboring a coding sequence for glutathione-S transferase (GST)-porcine ALS (pALS) fusion protein were expressed in BL21(DE3) E. coli and Sf9 insect cells, respectively. The expressed protein was purified by glutathione or Ni-NTN affinity chromatography, followed by cleavage of the fusion protein using Factor Xa. In addition, pALS and hIGFBP-3 were also produced in small amounts in the Xenopus oocyte expression system which does not require any purification procedure. A 65-kDa pALS polypeptide was obtained following the prokaryotic expression and the enzymatic digestion, but biochemical characterization of this polypeptide was precluded because of an extremely low expression efficiency. The baculovirus-as well as Xenopus-expressed pALS exhibited the expected molecular mass of 85 kDa which was reduced into 75 and 65 kDa following deglycosylation of Asn-linked carbohydrates by Endo-F glycosidase, indicating that the expressed pALS was properly glycosylated. Moreover, irrespective of the source of pALS, the recombinant pALS and hIGFBP-3 formed a 130-kDa binary complex which could be immunoprecipitated by anti-hIGFBP-3 antibodies. Collectively, results indicate that an authentic pALS protein can be produced by the current expression systems.

Establishment of a Binding Assay System for Screening of the Inhibitors of $p56^{lck}$ SH2 Domain

  • Kim, Jyn-Ho;Hur, Eun-Mi;Yun, Yung-Dae
    • BMB Reports
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    • v.31 no.4
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    • pp.370-376
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    • 1998
  • Src-Homology 2 (SH2) domains have a capacity to bind phosphotyrosine-containing sequence context and play essential roles in various cellular signaling pathways. Due to the specific nature of the binding between SH2 domains and their counterpart proteins, inhibitors of SID domain binding have drawn extensive attention as a potential candidate for therapeutic agents. Here, we describe the binding assay system to screen for the ligands or blockers of the SH2 domains with an emphasis on the $p56^{lck}$ SH2 domain. In our assay system, SID domains expressed and purified as fusion proteins to Glutathione-S-transferase (GST) were covalently attached to 96-well microtitre plates through amide bond formation, which were subsequently allowed to bind the biotinylated phosphotyrosine (pY)containing synthetic pep tides. The binding of biotinylated pY peptides was detected by the horseradish peroxidase (HRP)-conjugated streptavidin. Using the various combinations of SH2 domain-pY peptides, we observed that: (1) The binding of pY-peptides to its counterpart SH2 domain is concentration-dependent and saturable; (2) The binding is highly specific for a particular combination of SH2 domain-pY peptide pair; and (3) The binding of Lck SH2-cognate pY-peptides is specifically competed by the nonbiotinylated peptides with expected relative affinity. These results indicate that the established assay system detects the SH2-pY peptide interaction with reproducible sensitivity and specificity and is suitable for screening the specific inhibitors of $p56^{lck}$ SH2 function.

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