diseases pathogenesis are genes that encodes the synthesis of glutathione-Stransferase(GST),known as the second phase enzyme detoxification system that protects against endogenous oxidative stress and exogenous toxins...diseases pathogenesis are genes that encodes the synthesis of glutathione-Stransferase(GST),known as the second phase enzyme detoxification system that protects against endogenous oxidative stress and exogenous toxins,through catalisation of glutathione sulfuric groups conjugation and decontamination of lipid and deoxyribonucleic acid oxidation products.The group of GST enzymes consists of cytosolic,mitochondrial and microsomal fractions.Recently,eight classes of soluble cytoplasmic isoforms of GST enzymes are widely known:α-,ζ-,θ-,κ-,μ-,π-,σ-,andω-.The GSTs gene family in the Human Gene Nomenclature Committee,online database recorded over 20 functional genes.The level of GSTs expression is considered to be a crucial factor in determining the sensitivity of cells to a broad spectrum of toxins.Nevertheless,human GSTs genes have multiple and frequent polymorphisms that include the complete absence of the GSTM1 or the GSTT1 gene.Current review supports the position that genetic polymorphism of GST genes is involved in the pathogenesis of various liver diseases,particularly non-alcoholic fatty liver disease,hepatitis and liver cirrhosis of different etiology and hepatocellular carcinoma.Certain GST allelic variants were proven to be associated with susceptibility to hepatological pathology,and correlations with the natural course of the diseases were subsequently postulated.展开更多
The article presented the results of comparison of polymorphic variants of the genes GSTM1, GSTT1, GSTP1 and clinical manifestations of non-small cell lung carcinoma. The association of the genotype GSTT1 (del) with t...The article presented the results of comparison of polymorphic variants of the genes GSTM1, GSTT1, GSTP1 and clinical manifestations of non-small cell lung carcinoma. The association of the genotype GSTT1 (del) with the risk of developing squamous cell lung cancer has been revealed (OR = 2.54 CI: 1.13 - 5.72, p = 0.035). Analysis of patient survival rate (n = 173) in groups of various histological types of lung cancer showed that in the group of squamous cell lung cancer (n = 91) in patients with genotype GSTT1 (del), the survival rate median was significantly higher—84 months (95% CI 12.4 - 155.7) than in patients with the genotype GSTT1 (+)—36 months (95% CI 25.2 - 46.8, p = 0.045). In contrast, in the adenocarcinoma group (n = 82), the survival rate median in patients with the genotype GSTT1 (del) was 19 months. (95% CI 6.2 - 33.5), and in patients with genotype GSTT1 (+)—67 months (95% CI 50.1 - 84.0), which is the basis for continuing this comparison in an additional group of testees, as the sampling did not achieve the reliability of p = 0.12. Hypothetically, these differences may be due to differences in the gender composition of squamous cell lung cancer and adenocarcinoma and the involvement of GST enzymes in the metabolism of estrogens in adenocarcinoma in women and other hormonal background and reactivity of the male body with squamous cell carcinoma. Further research and subsequent analysis of the results will be aimed at confirming this hypothesis.展开更多
Periodontitis, is an infectious ailment of multifactorial origin, that brings about destruction of bone and surrounding tissues. There are various oral pathogens that may be responsible for the destruction. The host e...Periodontitis, is an infectious ailment of multifactorial origin, that brings about destruction of bone and surrounding tissues. There are various oral pathogens that may be responsible for the destruction. The host encounters these microbial invasions and their products by the production and release of inflammatory mediators from the cells within the body. Glutathione-S-transferase (GST) are a group of enzymes that utilize glutathione in conditions resulting in oxidative stress. These enzymes play a key role in the detoxifycation of such substance. It aids in preventing damage to important cellular components caused by release of free reactive oxygen species. Ceruloplasmin is a ferroxidase enzyme. It plays a role as an anti-inflammatory agent, by its ability to scavenge free radicals within the body. The present study was targeted at evaluating the levels of Glutathione-S-Transferase (GST) and Ceruloplasmin as diagnostic markers for patients with chronic periodontitis in gingival crevicular fluid (GCF) and the gingival tissues. Thirty patients were divided into two groups. Experimental group comprising of 15 subjects with chronic perio- dontitis and the control group was composed of 15 healthy individuals. Highly significant changes in GST between the diseased and normal patients (P = 0.001) were detected. There was a decrease in GST level in both gingival tissue & GCF in diseased patients when compared to the control patients. The ceruloplasmin levels in GCF and gingival tissues showed no difference between the control and diseased group. Hence,these results indicate a relationship suggesting that GST produced during chronic inflammation could be used as biomarker that indicate periodontal disease .展开更多
Objective:To elucidates the immunoprophylactic potential of glutathion-s-transferase(GST) from cattle filarial parasite Setaria digitata(S.digitata) against lymphatic filariasis.Methods: GST was purified through affin...Objective:To elucidates the immunoprophylactic potential of glutathion-s-transferase(GST) from cattle filarial parasite Setaria digitata(S.digitata) against lymphatic filariasis.Methods: GST was purified through affinity chromatography(SdGST) and chacterized by SDS-PAGE and Nano-LC MS/MS analysis.Antibody isotypes to SdGST were measured by ELISA.Antibody dependant cellular cytotoxicity(ADCC) was performed in vitro using sera from immunized animals and immune individuals.T-cell proliferation and cytokine response to SdGST in different groups of filariasis were measured.Immunoprophylactic potential of SdGST was evaluate in animal model.Results:SdGST exhibited 30-fold enhancement of enzyme activity over crude parasitic extract.It was found to be 26 kDa by SDS-PAGE.Nano LC-MS/MS analysis followed by blast search showed 100%homology with Dirqfilaria immitis(D.immitis) and only 43%with Homo sapiens(H.sapiens).Immunoblotting analysis showed putatively immune individuals carry significant level of antibodies to SdGST as compared with microfilaraemics.Immunized sera and sera endemic normal could neutralize the enzymatic activity of SdGST and inducing in vitro cytotoxicity of microfilariae.Peripheral blood mononuclear cells(PBMC) from endemic normals upon stimulation with SdGST showed a mixed type of Th1/Th2 response.SdGST immunization clear microfilariae from circulation in S.digitata implanted mastomys.Conclusions:The heterologous GST could be potentially developed as a vaccine candidate against lymphatic filarial parasite.展开更多
The glutathione-S-transferase genes mainly the GSTM1 and GSTT1 alleles are responsible for the synthesis of detoxication enzymes that can remove toxic substances. The objective of this study is to seek changes in the ...The glutathione-S-transferase genes mainly the GSTM1 and GSTT1 alleles are responsible for the synthesis of detoxication enzymes that can remove toxic substances. The objective of this study is to seek changes in the genetic polymorphism of glutathione-S-transferase GSTM1 and GSTT1 in motorcycle drivers exposed to BTEX. Our study group consists of 60 motorcycle drivers including 30 professional and 30 non-professional. Blood samples were preleveled from the study population in the EDTA tubes and DNA was extracted by the phenol/chloroform method. The PCR technique was used to determine the presence or absence of genes. Our results showed that the percentage of GSTM1 null genotype has a statistically significant difference (P = 0.02), while the percentage of GSTT1 null genotype was non-significant (P = 0.76) between the two groups. The percentage of deletion of both genes is higher in professional than non-professional motorcycle drivers. Air pollution in Cotonou by BTEX seems to influence the deletion of the GSTM1 and GSTT1 genes at a higher percentage among professional than non-professional motorcycle drivers.展开更多
Objective: In this study, we plan to measure plasma Catalase (CAT), Antioxidant Activity (AOA) and Glu- tathione-S-Transferase (GST) levels to understand whether oxidative stress develops or not and whether or not the...Objective: In this study, we plan to measure plasma Catalase (CAT), Antioxidant Activity (AOA) and Glu- tathione-S-Transferase (GST) levels to understand whether oxidative stress develops or not and whether or not the detoxification mechanism properly functions in children with Attention Deficit and Hyperactivity Disorder (ADHD) with unknown etiology and pathogenesis. Method and Results: Plasma CAT, AOA, and GST activities were spectrophotometrically measured in forty patients (average age 10.27 ± 2.54) and thirty-five (average age, 9.97 ± 2.59) healthy individuals as the control group. While the CAT activity showed no difference in the patient group (P > 0.05) compared to the control group, AOA and GST levels were found significantly meaningful (P = 0.001). Conclusion: In this pilot study ,the study shows that no oxidative stress develops in individuals with ADHD in high AOA and stable CAT activity, and that the de- toxification mechanism functions extremely in high GST activity. These findings need to be supported by other studies.展开更多
Objective:To evaluate the acaricidal activity of extracts of three essential oils of chamomile,marjoram and Eucalyptwi against Tetranychus urtiaie(T.urticae.)Koch.Methods:Extracts of three essential oils of chamomile,...Objective:To evaluate the acaricidal activity of extracts of three essential oils of chamomile,marjoram and Eucalyptwi against Tetranychus urtiaie(T.urticae.)Koch.Methods:Extracts of three essential oils of chamomile,marjoram and Eucalyptus with different concentrations(0.5%,1.0%,2.0%,3.0%and 4.0%)were used to control T.urticae Koch.Results:The results showed that chamomile(Chamomilla recutita)represented the most potent efficient acaricidal agent against Tetranychus followed by marjoram(Marjorana hortensis)and Eucalyptus.The LC_(50)values of chamomile,marjoram and Eucalyptus for adults were 0.65,1.84 and 2.18,respectively and for eggs 1.17,6.26 and 7.33,respectively.Activities of enzymes including glutathione-Stxansferase,esterase(α-esterase andβ-esterase)and alkaline phosphatase in susceptible mites were determined and activities of enzymes involved in the resistance of acaricides were proved.Protease enzyme was significantly decreased at LC_(50)of both chamomile and marjoram compared with positive control.Gas chromatography-mass spectrometer(CC-MS)proved that the major compositions of Chamomilla recutita areα-bisabolol oxide A(35.251%),and trans-3-farersene(7.758%),while the main components of Marjorana hortensis are terpincne-4-ol(23.860%),p-cymene(23.404%)and sabinene(10.904%).Conclusions:It can be concluded that extracts of three essential oils of chamomile,marjoram and Eucalyptus possess acaricidal activity against T.urticae.展开更多
The aim of this study is to construct a prokaryotic expression vector of mouse Nanog gene and to express it in E. coli. A pair of primers was designed according to digestion sites in plasmid pGEX-KG and the Nanog gene...The aim of this study is to construct a prokaryotic expression vector of mouse Nanog gene and to express it in E. coli. A pair of primers was designed according to digestion sites in plasmid pGEX-KG and the Nanog gene sequence published by GenBank. The DNA fragment of 918 bp was amplified by polymerase chain reaction (PCR) from the pNA992 recombinant plasmid with Nanog gene, then cloned into pGEX-KG and transformed into the host E. coli strain TG Ⅰ. The sequence of the fragment was matched with the original sequence of pNA992. It indicated that fusion expression vector, pGEX-KG- Nanog, was constructed successfully. The pGEX-KG-Nanog plasmid was extracted from E. coli strain TG Ⅰ and was transformed into BL21(DE3) for expression. After induction by isopropyl-β-D-thiogalactoside (IPTG) at 37℃, the expression product of Nanog gene was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the expression condition was optimized. Nanog fusion protein was successfully expressed in the form of inclusion bodies. The molecular weight of the inclusion body was 63 kDa. Meanwhile, the optimum condition for the expression of Nanog fusion protein was induced with 0.8 mmol L^-1 IPTG for 5 h. The mouse Nanog gene was successfully expressed in E. coli, which laid a foundation for the purification of Nanog protein and for the preparation of polyclonal antibody.展开更多
AIM: To reveal the expression of multidrug-resistance associated proteins: glutathione-S-transferase π(GSTπ), P-glycoprotein(P-gp) and vault protein lung resistance protein(LRP) in retinoblastoma(RB) witho...AIM: To reveal the expression of multidrug-resistance associated proteins: glutathione-S-transferase π(GSTπ), P-glycoprotein(P-gp) and vault protein lung resistance protein(LRP) in retinoblastoma(RB) without any conservative treatment before primary enucleation and to correlate this expression with histopathological tumor features. METHODS: A total of 42 specimens of RB undergone primary enucleation were selected for the research. Sections from the formalin-fixed, paraffin-embedded specimens were stained with HE and immunohistochemistry to detect the expression of GSTπ, P-gp and LRP.RESULTS: GSTπ was expressed in 39/42(92.86%) RBs and in 9/9(100%) well-differentiated RBs. P-gp/GSTπ was found in 30(71.42%) of 42 RBs. Totally 9(21.43%) tumors were well differentiated and 33(78.57%) were poorly differentiated. Totally 15(35.71%) eyes had optic nerve(ON) tumor invasion, 36(85.71%) had choroidal tumor invasion, and 14(33.33%) had simultaneous choroidal and ON invasion. There was no statistically significant relationship between P-gp, GSTπ, LRP positivity and the degree of ocular layer tumor invasion and ON tumor invasion(P〉0.05). CONCLUSION: RB intrinsically expresses GSTπ, P-gp and LRP. GSTπ expression is positive in 100% welldifferentiation ones, so in which way it is correlated with differentiation. But the other two proteins expressions are not related to tumor differentiation and to the degree of tumor invasion. GSTπ may be a new target of chemotherapy in RB.展开更多
Although oxidation is the most common biological and energy producing reaction, oxidative stress is harmful to cell, because the products of oxidation such as free radicals and peroxides damage the cellular components...Although oxidation is the most common biological and energy producing reaction, oxidative stress is harmful to cell, because the products of oxidation such as free radicals and peroxides damage the cellular components, causing several diseases. Damage in DNA is responsible for cancer formation and progression. However, several enzymes such as superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase etc. act as antioxidants to influence oxidative stress. Polymorphisms in these enzymes are supposed to be associated with DNA damage and subsequently the individual's risk of cancer susceptibility. This review article aims to further elucidate the relationship between antioxidant enzymes and cancers by summarizing the findings of some of the important study concerning expression levels and genetic polymorphisms of antioxidant enzymes in cancer patients.展开更多
Objective:To evaluate in ritro and in riro antioxidant potency of Pistacia chinensis(P.chinensis) bark and leaves extracts along with its protective role against CCl_4 induced toxicity in testis of the rat.Methods Var...Objective:To evaluate in ritro and in riro antioxidant potency of Pistacia chinensis(P.chinensis) bark and leaves extracts along with its protective role against CCl_4 induced toxicity in testis of the rat.Methods Various in vitro models such as DPPH,ABTS,hydrogen peroxide,superoxide,hydroxyl and nitric oxide scavenging activities,anti-lipid peroxidation activity,phospho-molybdenum activity.β carotene bleaching assay was used for analysis of antioxidant potential.Experimental groups for in riro study were:Group Ⅰ(control)untreated.Group Ⅱ(Vehicle control).Group Ⅲ(1 mL/kg b.w 30%CCl_4).Group Ⅳ(1 mL/kg b.w CCk_4+Silymarin).Group Ⅴ(200 mg/kg b.w PCBE+CCl_4),Group Ⅵ(400 mg/kg b.w PCBE+CCl_4) and Group Ⅶ(400 mg/kg b.w PCBE alone).Results:In ritro antioxidant assays displayed significant results and the highest activity was not specified to a specific extract.However,ethyl acetate extract of bark(PCBF) showed highest results in most of the antioxidant assays i.e.beta-carotene bleaching,hydroxyl radical scavenging.ABTS.lipid peroxidation and superoxide radical scavenging activity.On this base,this fraction was selected for in riro antioxidant experiment.Testis tissues were analyzed to observe the protective effects of PCBE on antioxidant enzymes:cataluse,superoxide dismutase.peroxidase.glutathione-Stransferase.glutathione reductase,glutathione peroxidase and quinone reductase activities and glutathione(GSH) as well as nitrite content.Profile of plasma testosterone was also compared to various treatments.Observation suggests a protective role of P.chinensis against CCl_4induced toxicity.Conclusions:It is concluded that some bioactive antioxidants of P.chinensis bark might be a good source to isolate the potent antioxidant components.展开更多
Multidrug resistance (MDR) is a critical problem in cancer chemotherapy. Cancer cells can develop resistance not only to a single cytotoxic drug, but also to entire classes of structurally and functionally unrelated c...Multidrug resistance (MDR) is a critical problem in cancer chemotherapy. Cancer cells can develop resistance not only to a single cytotoxic drug, but also to entire classes of structurally and functionally unrelated compounds. Several mechanisms can mediate the development of MDR, including increased drug efflux from the cells by ABC-transporters (ABCT), activation of metabolic enzymes, and defective pathways towards apoptosis. Many plant secondary metabolites (SMs) can potentially increase sensitivity of drug-resistant cancer cells to chemotherapeutical agents. The present thesis investigates the modulation of MDR by certain medicinal plants and their active compounds. The inhibition of ABCTs (P-gp/MDR1, MRP1, BCRP) and metabolic enzymes (GST and CYP3A4), and the induction of apoptosis are useful indicators of the efficacy of a potential medicinal drug. The focus of this study was the possible mechanisms of drug resistance including: expression of resistance proteins, activation of metabolic enzymes, and alteration of the apoptosis and how to overcome their resistance effect on cancer cells. The overall goal of this review was to evaluate how commonly used medicinal plants and their main active secondary metabolites modulate multidrug resistance in cancer cells in order to validate their uses as anticancer drugs, introduce new therapeutic options for resistant cancer, and facilitate the development of their anticancer strategies and/or combination therapies. In conclusion, SMs from medicinal plants exhibit multitarget activity against MDR-related proteins, metabolic enzymes, and apoptotic signaling, this may help to overcome resistance towards chemotherapeutic drugs.展开更多
A comparative study of just cadmium (Cd) or heat and their combination treatments on some physiological parameters and the antioxidant systems in transgenic rice (Oryza sativa L. cv. Zhonghua No.11) carrying gluta...A comparative study of just cadmium (Cd) or heat and their combination treatments on some physiological parameters and the antioxidant systems in transgenic rice (Oryza sativa L. cv. Zhonghua No.11) carrying glutathione-S-transferase (GST, EC. 2.5.1.18) and catalasel (CAT1, EC. 1.11.1.6) and non-transgenics was conducted. The results revealed improved resistance in the transgenics to Cd and the combined Cd and heat stress than non-transgenics. Data showed that the activities of CAT, GST, superoxide dismutase (EC.1.15.1.1) and all components of the ascorbate-glutatbione cycle measured in the stressed transgenics shoots are significantly different from those of non-transgenics. Results indicated that co-expression of GST and CAT1 had an important effect on the antioxidant system, in particular, the whole ascorbate-glutathione cycle. The less oxidative damage induced by Cd and the stress combination in the transgenics resulted not only from the GST and CAT1 transgene but also from the coordination of the whole ascorbate-glutathione cycle.展开更多
Objective To investigate the effects of herbal compound 861 (Cpd861) on hepatocarcinogenesis induced by diethylntrosamine and 2-acetylaminofluorene (DEN-AAF) in female Sprague Dawley rats.Methods Liver preneoplastic f...Objective To investigate the effects of herbal compound 861 (Cpd861) on hepatocarcinogenesis induced by diethylntrosamine and 2-acetylaminofluorene (DEN-AAF) in female Sprague Dawley rats.Methods Liver preneoplastic foci were induced using the DEN-AAF method in female Sprague Dawley rats, which were then treated with Cpd861. For quantitative assessment of liver preneoplastic foci, the placental form of glutathione-S-transferase (GST-P) positive foci were measured using immunohistochemical staining and image analysis. GST-P protein expression was measured by Western blotting, mRNA expression was assessed by Northern blotting.Results Treatment using DEN-AAF caused a significant decrease in body weight and increase in liver weight compared to the control group. Oral Cpd861 administration essentially prevented DEN-AAF-induced body weight loss and liver weight increase. When 2-AAF was followed by treatment with Cpd861, there was a decrease in the number of large foci as compared to 2-AAF alone. However, there were still considerable numbers of small mixed clear/vacuolated cell foci, some of which were positive for GST-P. Significant increase in GST-P protein and mRNA expression were observed in the DEN-AAF group, while treatment with Cpd861 inhibited the increase. The effect of Cpd861 on hepatocarcinogenesis occurred in a concentration-dependent manner. Conclusion Chinese herbal compound Cpd861 prevents hepatocarcinogenesis in DEN-AAF-induced liver preneoplastic lesions in rats.展开更多
文摘diseases pathogenesis are genes that encodes the synthesis of glutathione-Stransferase(GST),known as the second phase enzyme detoxification system that protects against endogenous oxidative stress and exogenous toxins,through catalisation of glutathione sulfuric groups conjugation and decontamination of lipid and deoxyribonucleic acid oxidation products.The group of GST enzymes consists of cytosolic,mitochondrial and microsomal fractions.Recently,eight classes of soluble cytoplasmic isoforms of GST enzymes are widely known:α-,ζ-,θ-,κ-,μ-,π-,σ-,andω-.The GSTs gene family in the Human Gene Nomenclature Committee,online database recorded over 20 functional genes.The level of GSTs expression is considered to be a crucial factor in determining the sensitivity of cells to a broad spectrum of toxins.Nevertheless,human GSTs genes have multiple and frequent polymorphisms that include the complete absence of the GSTM1 or the GSTT1 gene.Current review supports the position that genetic polymorphism of GST genes is involved in the pathogenesis of various liver diseases,particularly non-alcoholic fatty liver disease,hepatitis and liver cirrhosis of different etiology and hepatocellular carcinoma.Certain GST allelic variants were proven to be associated with susceptibility to hepatological pathology,and correlations with the natural course of the diseases were subsequently postulated.
文摘The article presented the results of comparison of polymorphic variants of the genes GSTM1, GSTT1, GSTP1 and clinical manifestations of non-small cell lung carcinoma. The association of the genotype GSTT1 (del) with the risk of developing squamous cell lung cancer has been revealed (OR = 2.54 CI: 1.13 - 5.72, p = 0.035). Analysis of patient survival rate (n = 173) in groups of various histological types of lung cancer showed that in the group of squamous cell lung cancer (n = 91) in patients with genotype GSTT1 (del), the survival rate median was significantly higher—84 months (95% CI 12.4 - 155.7) than in patients with the genotype GSTT1 (+)—36 months (95% CI 25.2 - 46.8, p = 0.045). In contrast, in the adenocarcinoma group (n = 82), the survival rate median in patients with the genotype GSTT1 (del) was 19 months. (95% CI 6.2 - 33.5), and in patients with genotype GSTT1 (+)—67 months (95% CI 50.1 - 84.0), which is the basis for continuing this comparison in an additional group of testees, as the sampling did not achieve the reliability of p = 0.12. Hypothetically, these differences may be due to differences in the gender composition of squamous cell lung cancer and adenocarcinoma and the involvement of GST enzymes in the metabolism of estrogens in adenocarcinoma in women and other hormonal background and reactivity of the male body with squamous cell carcinoma. Further research and subsequent analysis of the results will be aimed at confirming this hypothesis.
文摘Periodontitis, is an infectious ailment of multifactorial origin, that brings about destruction of bone and surrounding tissues. There are various oral pathogens that may be responsible for the destruction. The host encounters these microbial invasions and their products by the production and release of inflammatory mediators from the cells within the body. Glutathione-S-transferase (GST) are a group of enzymes that utilize glutathione in conditions resulting in oxidative stress. These enzymes play a key role in the detoxifycation of such substance. It aids in preventing damage to important cellular components caused by release of free reactive oxygen species. Ceruloplasmin is a ferroxidase enzyme. It plays a role as an anti-inflammatory agent, by its ability to scavenge free radicals within the body. The present study was targeted at evaluating the levels of Glutathione-S-Transferase (GST) and Ceruloplasmin as diagnostic markers for patients with chronic periodontitis in gingival crevicular fluid (GCF) and the gingival tissues. Thirty patients were divided into two groups. Experimental group comprising of 15 subjects with chronic perio- dontitis and the control group was composed of 15 healthy individuals. Highly significant changes in GST between the diseased and normal patients (P = 0.001) were detected. There was a decrease in GST level in both gingival tissue & GCF in diseased patients when compared to the control patients. The ceruloplasmin levels in GCF and gingival tissues showed no difference between the control and diseased group. Hence,these results indicate a relationship suggesting that GST produced during chronic inflammation could be used as biomarker that indicate periodontal disease .
基金financial support and to the Director RMRC,Bhubaneswar for providing necessary facilities for the study
文摘Objective:To elucidates the immunoprophylactic potential of glutathion-s-transferase(GST) from cattle filarial parasite Setaria digitata(S.digitata) against lymphatic filariasis.Methods: GST was purified through affinity chromatography(SdGST) and chacterized by SDS-PAGE and Nano-LC MS/MS analysis.Antibody isotypes to SdGST were measured by ELISA.Antibody dependant cellular cytotoxicity(ADCC) was performed in vitro using sera from immunized animals and immune individuals.T-cell proliferation and cytokine response to SdGST in different groups of filariasis were measured.Immunoprophylactic potential of SdGST was evaluate in animal model.Results:SdGST exhibited 30-fold enhancement of enzyme activity over crude parasitic extract.It was found to be 26 kDa by SDS-PAGE.Nano LC-MS/MS analysis followed by blast search showed 100%homology with Dirqfilaria immitis(D.immitis) and only 43%with Homo sapiens(H.sapiens).Immunoblotting analysis showed putatively immune individuals carry significant level of antibodies to SdGST as compared with microfilaraemics.Immunized sera and sera endemic normal could neutralize the enzymatic activity of SdGST and inducing in vitro cytotoxicity of microfilariae.Peripheral blood mononuclear cells(PBMC) from endemic normals upon stimulation with SdGST showed a mixed type of Th1/Th2 response.SdGST immunization clear microfilariae from circulation in S.digitata implanted mastomys.Conclusions:The heterologous GST could be potentially developed as a vaccine candidate against lymphatic filarial parasite.
文摘The glutathione-S-transferase genes mainly the GSTM1 and GSTT1 alleles are responsible for the synthesis of detoxication enzymes that can remove toxic substances. The objective of this study is to seek changes in the genetic polymorphism of glutathione-S-transferase GSTM1 and GSTT1 in motorcycle drivers exposed to BTEX. Our study group consists of 60 motorcycle drivers including 30 professional and 30 non-professional. Blood samples were preleveled from the study population in the EDTA tubes and DNA was extracted by the phenol/chloroform method. The PCR technique was used to determine the presence or absence of genes. Our results showed that the percentage of GSTM1 null genotype has a statistically significant difference (P = 0.02), while the percentage of GSTT1 null genotype was non-significant (P = 0.76) between the two groups. The percentage of deletion of both genes is higher in professional than non-professional motorcycle drivers. Air pollution in Cotonou by BTEX seems to influence the deletion of the GSTM1 and GSTT1 genes at a higher percentage among professional than non-professional motorcycle drivers.
文摘Objective: In this study, we plan to measure plasma Catalase (CAT), Antioxidant Activity (AOA) and Glu- tathione-S-Transferase (GST) levels to understand whether oxidative stress develops or not and whether or not the detoxification mechanism properly functions in children with Attention Deficit and Hyperactivity Disorder (ADHD) with unknown etiology and pathogenesis. Method and Results: Plasma CAT, AOA, and GST activities were spectrophotometrically measured in forty patients (average age 10.27 ± 2.54) and thirty-five (average age, 9.97 ± 2.59) healthy individuals as the control group. While the CAT activity showed no difference in the patient group (P > 0.05) compared to the control group, AOA and GST levels were found significantly meaningful (P = 0.001). Conclusion: In this pilot study ,the study shows that no oxidative stress develops in individuals with ADHD in high AOA and stable CAT activity, and that the de- toxification mechanism functions extremely in high GST activity. These findings need to be supported by other studies.
基金Supported by Cairo University,Faculty of Agriculture,Deptarment of Biochemistry,Cairo,Egypt
文摘Objective:To evaluate the acaricidal activity of extracts of three essential oils of chamomile,marjoram and Eucalyptwi against Tetranychus urtiaie(T.urticae.)Koch.Methods:Extracts of three essential oils of chamomile,marjoram and Eucalyptus with different concentrations(0.5%,1.0%,2.0%,3.0%and 4.0%)were used to control T.urticae Koch.Results:The results showed that chamomile(Chamomilla recutita)represented the most potent efficient acaricidal agent against Tetranychus followed by marjoram(Marjorana hortensis)and Eucalyptus.The LC_(50)values of chamomile,marjoram and Eucalyptus for adults were 0.65,1.84 and 2.18,respectively and for eggs 1.17,6.26 and 7.33,respectively.Activities of enzymes including glutathione-Stxansferase,esterase(α-esterase andβ-esterase)and alkaline phosphatase in susceptible mites were determined and activities of enzymes involved in the resistance of acaricides were proved.Protease enzyme was significantly decreased at LC_(50)of both chamomile and marjoram compared with positive control.Gas chromatography-mass spectrometer(CC-MS)proved that the major compositions of Chamomilla recutita areα-bisabolol oxide A(35.251%),and trans-3-farersene(7.758%),while the main components of Marjorana hortensis are terpincne-4-ol(23.860%),p-cymene(23.404%)and sabinene(10.904%).Conclusions:It can be concluded that extracts of three essential oils of chamomile,marjoram and Eucalyptus possess acaricidal activity against T.urticae.
文摘The aim of this study is to construct a prokaryotic expression vector of mouse Nanog gene and to express it in E. coli. A pair of primers was designed according to digestion sites in plasmid pGEX-KG and the Nanog gene sequence published by GenBank. The DNA fragment of 918 bp was amplified by polymerase chain reaction (PCR) from the pNA992 recombinant plasmid with Nanog gene, then cloned into pGEX-KG and transformed into the host E. coli strain TG Ⅰ. The sequence of the fragment was matched with the original sequence of pNA992. It indicated that fusion expression vector, pGEX-KG- Nanog, was constructed successfully. The pGEX-KG-Nanog plasmid was extracted from E. coli strain TG Ⅰ and was transformed into BL21(DE3) for expression. After induction by isopropyl-β-D-thiogalactoside (IPTG) at 37℃, the expression product of Nanog gene was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the expression condition was optimized. Nanog fusion protein was successfully expressed in the form of inclusion bodies. The molecular weight of the inclusion body was 63 kDa. Meanwhile, the optimum condition for the expression of Nanog fusion protein was induced with 0.8 mmol L^-1 IPTG for 5 h. The mouse Nanog gene was successfully expressed in E. coli, which laid a foundation for the purification of Nanog protein and for the preparation of polyclonal antibody.
基金Supported by the National Natural Science Foundation of China(No.30371515)
文摘AIM: To reveal the expression of multidrug-resistance associated proteins: glutathione-S-transferase π(GSTπ), P-glycoprotein(P-gp) and vault protein lung resistance protein(LRP) in retinoblastoma(RB) without any conservative treatment before primary enucleation and to correlate this expression with histopathological tumor features. METHODS: A total of 42 specimens of RB undergone primary enucleation were selected for the research. Sections from the formalin-fixed, paraffin-embedded specimens were stained with HE and immunohistochemistry to detect the expression of GSTπ, P-gp and LRP.RESULTS: GSTπ was expressed in 39/42(92.86%) RBs and in 9/9(100%) well-differentiated RBs. P-gp/GSTπ was found in 30(71.42%) of 42 RBs. Totally 9(21.43%) tumors were well differentiated and 33(78.57%) were poorly differentiated. Totally 15(35.71%) eyes had optic nerve(ON) tumor invasion, 36(85.71%) had choroidal tumor invasion, and 14(33.33%) had simultaneous choroidal and ON invasion. There was no statistically significant relationship between P-gp, GSTπ, LRP positivity and the degree of ocular layer tumor invasion and ON tumor invasion(P〉0.05). CONCLUSION: RB intrinsically expresses GSTπ, P-gp and LRP. GSTπ expression is positive in 100% welldifferentiation ones, so in which way it is correlated with differentiation. But the other two proteins expressions are not related to tumor differentiation and to the degree of tumor invasion. GSTπ may be a new target of chemotherapy in RB.
基金supported by the National Natural Science Foundation of China(No.30371660)the grant from the Chinese Medical Board(CMB) of New York(No.99-698).
文摘Although oxidation is the most common biological and energy producing reaction, oxidative stress is harmful to cell, because the products of oxidation such as free radicals and peroxides damage the cellular components, causing several diseases. Damage in DNA is responsible for cancer formation and progression. However, several enzymes such as superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase etc. act as antioxidants to influence oxidative stress. Polymorphisms in these enzymes are supposed to be associated with DNA damage and subsequently the individual's risk of cancer susceptibility. This review article aims to further elucidate the relationship between antioxidant enzymes and cancers by summarizing the findings of some of the important study concerning expression levels and genetic polymorphisms of antioxidant enzymes in cancer patients.
文摘Objective:To evaluate in ritro and in riro antioxidant potency of Pistacia chinensis(P.chinensis) bark and leaves extracts along with its protective role against CCl_4 induced toxicity in testis of the rat.Methods Various in vitro models such as DPPH,ABTS,hydrogen peroxide,superoxide,hydroxyl and nitric oxide scavenging activities,anti-lipid peroxidation activity,phospho-molybdenum activity.β carotene bleaching assay was used for analysis of antioxidant potential.Experimental groups for in riro study were:Group Ⅰ(control)untreated.Group Ⅱ(Vehicle control).Group Ⅲ(1 mL/kg b.w 30%CCl_4).Group Ⅳ(1 mL/kg b.w CCk_4+Silymarin).Group Ⅴ(200 mg/kg b.w PCBE+CCl_4),Group Ⅵ(400 mg/kg b.w PCBE+CCl_4) and Group Ⅶ(400 mg/kg b.w PCBE alone).Results:In ritro antioxidant assays displayed significant results and the highest activity was not specified to a specific extract.However,ethyl acetate extract of bark(PCBF) showed highest results in most of the antioxidant assays i.e.beta-carotene bleaching,hydroxyl radical scavenging.ABTS.lipid peroxidation and superoxide radical scavenging activity.On this base,this fraction was selected for in riro antioxidant experiment.Testis tissues were analyzed to observe the protective effects of PCBE on antioxidant enzymes:cataluse,superoxide dismutase.peroxidase.glutathione-Stransferase.glutathione reductase,glutathione peroxidase and quinone reductase activities and glutathione(GSH) as well as nitrite content.Profile of plasma testosterone was also compared to various treatments.Observation suggests a protective role of P.chinensis against CCl_4induced toxicity.Conclusions:It is concluded that some bioactive antioxidants of P.chinensis bark might be a good source to isolate the potent antioxidant components.
文摘Multidrug resistance (MDR) is a critical problem in cancer chemotherapy. Cancer cells can develop resistance not only to a single cytotoxic drug, but also to entire classes of structurally and functionally unrelated compounds. Several mechanisms can mediate the development of MDR, including increased drug efflux from the cells by ABC-transporters (ABCT), activation of metabolic enzymes, and defective pathways towards apoptosis. Many plant secondary metabolites (SMs) can potentially increase sensitivity of drug-resistant cancer cells to chemotherapeutical agents. The present thesis investigates the modulation of MDR by certain medicinal plants and their active compounds. The inhibition of ABCTs (P-gp/MDR1, MRP1, BCRP) and metabolic enzymes (GST and CYP3A4), and the induction of apoptosis are useful indicators of the efficacy of a potential medicinal drug. The focus of this study was the possible mechanisms of drug resistance including: expression of resistance proteins, activation of metabolic enzymes, and alteration of the apoptosis and how to overcome their resistance effect on cancer cells. The overall goal of this review was to evaluate how commonly used medicinal plants and their main active secondary metabolites modulate multidrug resistance in cancer cells in order to validate their uses as anticancer drugs, introduce new therapeutic options for resistant cancer, and facilitate the development of their anticancer strategies and/or combination therapies. In conclusion, SMs from medicinal plants exhibit multitarget activity against MDR-related proteins, metabolic enzymes, and apoptotic signaling, this may help to overcome resistance towards chemotherapeutic drugs.
基金Supported by the National Natural Science Foundation of China (30671126)
文摘A comparative study of just cadmium (Cd) or heat and their combination treatments on some physiological parameters and the antioxidant systems in transgenic rice (Oryza sativa L. cv. Zhonghua No.11) carrying glutathione-S-transferase (GST, EC. 2.5.1.18) and catalasel (CAT1, EC. 1.11.1.6) and non-transgenics was conducted. The results revealed improved resistance in the transgenics to Cd and the combined Cd and heat stress than non-transgenics. Data showed that the activities of CAT, GST, superoxide dismutase (EC.1.15.1.1) and all components of the ascorbate-glutatbione cycle measured in the stressed transgenics shoots are significantly different from those of non-transgenics. Results indicated that co-expression of GST and CAT1 had an important effect on the antioxidant system, in particular, the whole ascorbate-glutathione cycle. The less oxidative damage induced by Cd and the stress combination in the transgenics resulted not only from the GST and CAT1 transgene but also from the coordination of the whole ascorbate-glutathione cycle.
文摘Objective To investigate the effects of herbal compound 861 (Cpd861) on hepatocarcinogenesis induced by diethylntrosamine and 2-acetylaminofluorene (DEN-AAF) in female Sprague Dawley rats.Methods Liver preneoplastic foci were induced using the DEN-AAF method in female Sprague Dawley rats, which were then treated with Cpd861. For quantitative assessment of liver preneoplastic foci, the placental form of glutathione-S-transferase (GST-P) positive foci were measured using immunohistochemical staining and image analysis. GST-P protein expression was measured by Western blotting, mRNA expression was assessed by Northern blotting.Results Treatment using DEN-AAF caused a significant decrease in body weight and increase in liver weight compared to the control group. Oral Cpd861 administration essentially prevented DEN-AAF-induced body weight loss and liver weight increase. When 2-AAF was followed by treatment with Cpd861, there was a decrease in the number of large foci as compared to 2-AAF alone. However, there were still considerable numbers of small mixed clear/vacuolated cell foci, some of which were positive for GST-P. Significant increase in GST-P protein and mRNA expression were observed in the DEN-AAF group, while treatment with Cpd861 inhibited the increase. The effect of Cpd861 on hepatocarcinogenesis occurred in a concentration-dependent manner. Conclusion Chinese herbal compound Cpd861 prevents hepatocarcinogenesis in DEN-AAF-induced liver preneoplastic lesions in rats.