Lower Glutathione S-transferase Research Articles

A Glutathione S-Transferase from Thinopyrum ponticum Confers Fhb7 Resistance to Fusarium Head Blight in Wheat.

Fusarium head blight (FHB), mainly incited by Fusarium graminearum, has caused great losses in grain yield and quality of wheat globally. Fhb7, a major gene from 7E chromosome of Thinopyrum ponticum, confers broad resistance to multiple Fusarium species in wheat and has recently been cloned and identified as encoding a glutathione S-transferase (GST). However, some recent reports raised doubt about whether GST is the causal gene of Fhb7. To resolve the discrepancy and validate the gene function of GST in wheat, we phenotyped Fhb7 near-isogenic lines (Jimai22-Fhb7 versus Jimai22) and GST overexpressed lines for FHB resistance. Jimai22-Fhb7 showed significantly higher FHB resistance with a lower percentage of symptomatic spikelets, Fusarium-damaged kernels, and deoxynivalenol content than susceptible Jimai22 in three experiments. All the positive GST transgenic lines driven by either the maize ubiquitin promoter or its native promoter with high gene expression in the wheat cultivar 'Fielder' showed high FHB resistance. Only one maize ubiquitin promoter-driven transgenic line showed low GST expression and similar susceptibility to Fielder, suggesting that high GST expression confers Fhb7 resistance to FHB. Knockout of GST in the Jimai22-Fhb7 line using CRISPR-Cas9-based gene editing showed significantly higher FHB susceptibility compared with the nonedited control plants. Therefore, we confirmed GST as the causal gene of Fhb7 for FHB resistance. Considering its major effect on FHB resistance, pyramiding Fhb7 with other quantitative trait loci has a great potential to create highly FHB-resistant wheat cultivars.

Glutathione S-transferase activity facilitates rice tolerance to the barnyard grass root exudate DIMBOA

BackgroundIn paddy fields, the noxious weed barnyard grass secretes 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) to interfere with rice growth. Rice is unable to synthesize DIMBOA. Rice cultivars with high or low levels of allelopathy may respond differently to DIMBOA.ResultsIn this study, we found that low concentrations of DIMBOA (≤ 0.06 mM) promoted seedling growth in allelopathic rice PI312777, while DIMBOA (≤ 0.08 mM) had no significant influence on the nonallelopathic rice Lemont. DIMBOA treatment caused changes in the expression of a large number of glutathione S-transferase (GST) proteins, which resulting in enrichment of the glutathione metabolic pathway. This pathway facilitates plant detoxification of heterologous substances. The basal levels of GST activity in Lemont were significantly higher than those in PI312777, while GST activity in PI312777 was slightly induced by increasing DIMBOA concentrations. Overexpression of GST genes (Os09g0367700 and Os01g0949800) in these two cultivars enhanced rice resistance to DIMBOA.ConclusionsTaken together, our results indicated that different rice accessions with different levels of allelopathy have variable tolerance to DIMBOA. Lemont had higher GST activity, which helped it tolerate DIMBOA, while PI312777 had lower GST activity that was more inducible. The enhancement of GST expression facilitates rice tolerance to DIMBOA toxins from barnyard grass root exudates.

Distribution of metals in different environmental compartments and oxidative stress biomarkers in Bryconops caudomaculatus (Osteichthyes: Characiformes) from a bauxite mining area in the Eastern Amazon

The Eastern Amazon is rich in bauxite ore. The extraction and processing of bauxite lead to the mobilization of Aluminum (Al) and other metals in environmental. We evaluated the metals (Al, Mn, Ba, and Cr) concentration in tissue, water, and sediment associated with antioxidant and oxidative damage responses in Bryconops caudomaculatus. The samplings were done in two hydrological periods (post-rain and post-dry periods) and at three points, located at two rivers: one in the surroundings of the mining area (P1) and other inside the mining area, upstream (P2), and downstream (P3). Defense antioxidant system biomarkers analyzed were total antioxidant capacity (ACAP) and glutathione-S-transferase (GST) activity. As an oxidative damage biomarker, the lipoperoxidation (LPO) was evaluated. Metals concentrations in the water and sediment were higher in the post-rain period compared to post-dry period. The water samples were acidic, with dissolved Al concentrations above the values established by local legislation at all points. In the gills, the metals accumulation was higher in fish from in the surrounding and upstream sites, and in the liver, was higher in fish from downstream site. Fish from the surrounding had increased antioxidant defenses, with higher ACAP in all tissues and higher GST in the gills. Consequently, they had lower levels of LPO. Fish from the mining area had decreased antioxidant defenses, with lower ACAP in all tissues and lower GST in the gills. Consequently, they had higher levels of LPO, indicating oxidative stress. The fish muscle was not responsive to GST and LPO at all sites. We conclude that the oxidative stress observed in the gills and liver of B. caudomaculatus from the area modified by the mining activity reflected the local anthropogenic impact status.

Influence of 50 Hz electromagnetic frequency on oxidative stress and morphological characteristics in mosquito-borne filariasis Culex pipiens

• The larvae of Culex pipiens were exposed to EMF frequency (50 Hz). • The biochemical and morphological changes in Culex pipiens were estimated. • The larval body weight and total protein in EMF treated groups were significantly decreased compared to control. • Antioxidant enzymes activity in EMF treated groups were decreased significantly compared to the control. • Both 500 & 1000 mT caused distinct malformations to the larval body parts. The electromagnetic field (EMF) is a vital issue in research, but its use as an alternative technique for insect management is still in its beginnings. The study aimed to evaluate the effects of exposure to the electromagnetic field on the biochemical analysis and morphological characteristics in Culex pipiens . Therefore, the third instar larvae were exposed to EMF frequency (50 Hz) at different intensities (250, 500, 1000 mT) for 1 h during four successive days. After end exposure to EMF, the biochemical analysis, oxidative stress parameters, body weight, and morphological features were investigated. The results showed that larval body weight and total protein content were decreased significantly in all treated groups compared to controls. The total lipid content significantly decreased at 500 & 1000 mT exposure, while the treatment group exposed to EMF at 250 mT was not statistically different compared to control. At a high intensity of EMF 1000 mT all oxidative stress parameters; catalase (CAT) and Glutathione S-transferase (GST) activity and lipid peroxidation level (MDA) were decreased significantly compared to the control. On the other hand, at a low intensity (250 mT), CAT and GST activity was significantly increased, while MDA content was not statistically changed. Scanning electron microscopy showed that both 500 & 1000 mT caused distinct malformations to the larval body parts, mouthparts, thorax and abdomen with last abdominal structures. In conclusion, the results demonstrated that EMF exposure is a considerable stress factor that affects oxidative state and morphological features in mosquitoes and give hope that EMF will be used as an alternative method for Culex pipiens vector control.

MEASURE OF SERUM MDA, GST AND VIT-C IN WOMEN WITH CERVICAL CARCINOMA

Cervical carcinoma (CaCx) is the second most common gynecological cancers globally. Many authors revealed that low levels of antioxidants induce the overproduction of reactive oxygen species (ROS) that result in oxidative stress. In the present study an attempt was made to evaluate the oxidative stress in 20 women with cervical carcinoma by measuring serum Malondialdehyde (MDA), Glutathione-S-transferase (GST) and Ascorbic acid (Vit-C) levels which was compared with 20 age matched non cancerous women. Signicantly elevated MDA levels (P<0.001) and lowered GST, Vit-C levels (P<0.0001) were observed in women with cervical carcinoma as compared with healthy controls. From our ndings it can be concluded that oxidative stress plays an important role in pathogenesis of cervical carcinoma. Hence supplementation of antioxidants may reduce the oxidative stress and minimize the associated morbidity and mortality

Antioxidant balance in plasma of patients on home parenteral nutrition: A pilot study comparing three different lipid emulsions

Parenteral nutrition (PN) can supply all essential nutrients to a patient with gastrointestinal insufficiency. However, the sensitivity to lipid peroxidation might increase in those receiving PN, especially home parenteral nutrition (HPN). This study aimed to investigate whether PN affects the antioxidant balance of plasma of HPN patients without comorbidities and whether this balance is influenced by comorbidities and according to the type of lipid emulsion included in the PN. Adult patients on HPN (n=86) received one of three types of lipid emulsion (based on 1) soyabean oil, 2) olive and soyabean oil or 3) soyabean, coconut, olive and fish oil) in all-in-one mixtures; in addition healthy controls (n=66) were studied as comparators. HPN patients were classified to the following subgroups: 1) patients without (n=58) or with (n=28) comorbidities 2) patients on Intralipid (GINTRA, n=53), ClinOleic (GCLIN, n=17) or SMOFlipid (GSMOFn=16). The activities of total glutathione peroxidase (GSH-Px), selenium dependent glutathione peroxidase (Se-GSHPx) and glutathione S-transferase (GST) in plasma were determined spectrophotometrically. The antioxidant potential of plasma was determined using oxygen radical absorbance capacity (ORAC). The lipid peroxidation marker malondialdehyde (MDA) was analyzed with high performance liquid chromatography. MDA concentration was the highest in GINTRA and the lowest in GSMOF (p<0.05). GSMOF also had the highest activity of GSH-Px. No differences in Se-GSHPx, GST and ORAC were observed among GINTRA, GCLIN and GSMOF. Comparing with healthy controls, significantly lower GST (p=0.0293) and ORAC (p<0.0001) were observed in the HPN patients. Among all measured parameters only the concentration of MDA was significantly higher in patients with comorbidities compared to those without them. Comorbidities did not influence MDA level in GINTRA and GSMOF being still the lowest in GSMOF (p=0.0033). In contrast, significantly higher MDA level was observed for GCLIN in those with vs. without comorbidities (p=0.0262). Patients on HPN have lower antioxidant defenses than healthy controls. The type of lipid emulsion used in HPN affects lipid peroxidation (even after taking into account comorbidities which often involve oxidative stress) being the highest in GINTRA and the lowest in GSMOF. Thus, to minimize the risk of oxidative stress, SMOFlipid can be considered in patients in HPN especially for those with comorbidities. ClinOleic can be considered in HPN patients without comorbidities. The observation should be confirmed in larger studies.

Algae-derived polysaccharides supplementation ameliorates heat stress-induced impairment of bursa of Fabricius via modulating NF-κB signaling pathway in broilers

This study aimed to investigate the protective effects of dietary algae-derived polysaccharides (ADP) from Enteromorpha prolifera against heat stress (HS)-induced bursa of Fabricius injure in broilers, and to elucidate the molecular mechanisms underlying the protective effect. A total of 144 8-week-old male yellow-feathered broilers were randomly allocated into 3 treatments of 6 replicates each (8 broilers per replicate): thermoneutral zone group (TN, fed basal diet); heat stress group (HS, fed basal diet); heat stress + ADP group (HSA, basal diet supplemented with 1,000 mg/kg ADP). Broilers in TN group were raised at 23.6 ± 1.8°C during the whole study. Broilers in HS and HSA groups were exposed to 33.2 ± 1.5°C for 10 h/day. The experimental period lasted for four weeks. The results showed that HS and dietary ADP had no significant effects on bursa of Fabricius index (P > 0.05). HS exposure increased the apoptosis rate of bursa of Fabricius (P < 0.05), and the apoptosis rate was reduced by dietary ADP (P < 0.05). Besides, broilers in HS and HSA groups had a lower glutathione-S transferase (GST) activity and total anti-oxidation capacity (T-AOC), whereas had a higher malondialdehyde (MDA) levels of bursa of Fabricius than those in TN group (P < 0.05). HS exposure elevated the concentration of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-4, and IL-6, while decreased the concentration of interferon-γ (INF-γ) and IL-2 (P < 0.05), and dietary inclusion of ADP reduced the IL-1β level and increased the IL-2 level of bursa of Fabricius (P < 0.05). Compared with TN group, broilers in HS and HSA groups had lower relative mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1) and GSTT1 in bursa of Fabricius (P < 0.05). Additionally, HS exposure down-regulated the mRNA expression of inhibitor kappa B alpha (IκBα), IFN-γ, and IL-2, while up-regulated the mRNA expression of nuclear factor-kappa B (NF-κB) p65, TNF-α, IL-1β, and IL-6 in bursa of Fabricius (P < 0.05). However, dietary inclusion of ADP up-regulated the mRNA expression of IκBα and down-regulated the mRNA expression of NF-κB p65, TNF-α, and IL-6 in bursa of Fabricius (P < 0.05). Furthermore, HS exposure increased the relative protein expression levels of total and nuclear NF-κB p65 (P < 0.05), but dietary ADP supplementation reduced the relative protein expression levels of total and nuclear NF-κB p65 in bursa of Fabricius (P < 0.05). Collectively, dietary ADP ameliorated the impairment of histology, cell apoptosis and immune balance in bursa of Fabricius of heat stressed broilers, which is involved in modulation of NF-κB signaling pathway.

Gene expression profiling of reactive oxygen species (ROS) and antioxidant defense system following Sugarcane mosaic virus (SCMV) infection

BackgroundViruses are infectious pathogens, and plant virus epidemics can have devastating consequences to crop yield and quality. Sugarcane mosaic virus (SCMV, belonging to family Potyviridae) is one of the leading pathogens that affect the sugarcane crop every year. To combat the pathogens’ attack, plants generate reactive oxygen species (ROS) as the first line of defense whose sophisticated balance is achieved through well-organized antioxidant scavenging pathways.ResultsIn this study, we investigated the changes occurring at the transcriptomic level of ROS associated and ROS detoxification pathways of SCMV resistant (B-48) and susceptible (Badila) sugarcane genotypes, using Saccharum spontaneum L. genome assembly as a reference genome. Transcriptomic data highlighted the significant upregulation of ROS producing genes such as NADH oxidase, malate dehydrogenase and flavin-binding monooxygenase, in Badila genotype after SCMV pathogenicity. To scavenge the ROS, the Badila genotype illustrated a substantial enhancement of antioxidants i.e. glutathione s-transferase (GST), as compared to its resistant counterpart. GST is supposed to be a key indicator of pathogen attacks on the plant. A remarkably lower GST expression in B-48, as compared to Badila, indicated the development of resistance in this genotype. Additionally, we characterized the critical transcription factors (TFs) involved in endowing resistance to B-48. Among these, WRKY, AP2, NAC, bZIP, and bHLH showed enhanced expression in the B-48 genotype. Our results also confirmed the linkage of transcriptomic data with the enzymatic and qPCR data. The estimation of enzymatic activities for superoxide dismutase, catalase, ascorbate peroxidase, and phenylalanine ammonia-lyase supported the transcriptomic data and evinced higher resistance in B-48 genotype.ConclusionThe current study supported the efficiency of the B-48 genotype under SCMV infection. Moreover, comparative transcriptomic data has been presented to highlight the role of significant transcription factors conferring resistance to this genotype. This study provides an in-depth knowledge of the expression profiling of defense mechanisms in sugarcane.

Biomarkers of oxidative stress and cell damage in freshwater bivalves Diplodon parodizi exposed to landfill leachate.

Landfill is a public and environmental health problem; establishing and understanding methodologies to decrease its toxicity are thus necessary. Leachate samples were collected, at a sanitary landfill, immediately after the exit from the landfill, i.e. raw leachate (collection point A), after conventional treatment (point B) and after treatment by wetlands (point C). D. parodizi specimens were exposed to 3%, 10% and control (0%) dilutions of leachate from these collection points for 7days. Markers of antioxidant defences and cell damage were analysed. At point B, the gills of D. parodizi showed higher glutathione-S-transferase (GST) and glutathione reductase (GR) activity; the latter is a supplier of glutathione reductase (GSH). The low GST activity at point A was associated with the hormesis effect. Higher levels of superoxide dismutase (SOD), ethoxyresorufin-O-deethylase (EROD) and glutathione peroxidase (GPx) occurred at point A. Glucose-6-phosphate dehydrogenase (G6PDH) was inhibited at the points with the highest pollutant load and at the highest leachate dilutions. Higher levels of markers at point A may be related to the high pollutant charge and specific compounds present in the untreated leachate. The multi-xenobiotic resistance mechanism (MXR), metallothionein-like proteins (MT) and lipid peroxidation (LPO) did not vary among treatments. The biomarker responses showed negative effects of the leachate on the freshwater bivalve and simultaneously showed that the wetland treatment employed at the Caximba sanitary landfill is effective.

Lifetime Risk Factors for Pre- and Post-Bronchodilator Lung Function Decline. A Population-based Study.

Rationale: Interactions between early life and adult insults on lung function decline are not well understood, with most studies investigating prebronchodilator (pre-BD) FEV1 decline.Objectives: To investigate relationships between adult risk factors and pre- and post-BD lung function decline and their potential effect modification by early life and genetic factors.Methods: Multiple regression was used to examine associations between adult exposures (asthma, smoking, occupational exposures, traffic pollution, and obesity) and decline in both pre- and post-BD spirometry (forced expiratory volume in 1 s [FEV1], forced vital capacity [FVC], and FEV1/FVC) between ages 45 and 53 years in the Tasmanian Longitudinal Health Study (n = 857). Effect modification of these relationships by childhood respiratory risk factors, including low childhood lung function and GST (glutathione S-transferase) gene polymorphisms, was investigated.Results: Baseline asthma, smoking, occupational exposure to vapors/gases/dusts/fumes, and living close to traffic were associated with accelerated decline in both pre- and post-BD FEV1. These factors were also associated with FEV1/FVC decline. Occupational exposure to aromatic solvents was associated with pre-BD but not post-BD FEV1 decline. Maternal smoking accentuated the effect of personal smoking on pre- and post-BD FEV1 decline. Lower childhood lung function and having the GSTM1 null allele accentuated the effect of occupational exposure to vapors/gases/dusts/fumes and personal smoking on post-BD FEV1 decline. Incident obesity was associated with accelerated decline in FEV1 and more pronounced in FVC.Conclusions: This study provides new evidence for accentuation of individual susceptibility to adult risk factors by low childhood lung function, GSTM1 genotype, and maternal smoking.

Effects of pH on growth and biochemical responses in Agarophyton vermiculophyllum under different temperature conditions

The effects of pH (6.2, 7.2, 8.2, 9.2, and 10.2) under rising temperature (30 °C vs 20 °C) on Agarophyton vermiculophyllum growth and bio-physiology were investigated. Results showed that A. vermiculophyllum exhibited lower growth rates under elevated temperature in all pH values. Chlorophyll a, carotenoid, and phycocyanin levels were significantly enhanced by temperature elevation (p < 0.05). Enhanced H2O2 production either at lower or higher pH values correlated with lipid peroxidation (LPO) levels under elevated temperature, which suggested oxidative stress development. Oxidative damage was more severe at elevated pH values, which is confirmed by higher reactive oxygen species (ROS) levels. Compared with ambient pH 8.2 value, lower pH values under elevated temperature lead to increase activities of superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST), indicating that these enzymes played an important role to combat stress. However, decreased glutathione reductase (GR) and glutathione peroxidase (GPx) activities indicate least contribution for ROS scavenging at lower pH values. On contrary, SOD and CAT declined at elevated pH values compared with ambient pH, suggesting least contribution for ROS removal. Moreover, enhanced GR and GPx activities at elevated pH and temperature are not enough to scavenge ROS production. These data are consistent with higher H2O2 and LPO levels, and lower GST activities. Collectively, our results indicated that either pH fluctuations or elevated temperature displayed a disadvantageous influence on growth and bio-physiology of A. vermiculophyllum. Therefore, rising temperature alleviates adverse effects of seawater acidification, but it aggravates the negative effects of seawater alkalization on growth and bio-physiology of A. vermiculophyllum.

Human Adenovirus, Mesophilic Bacteria and Fungi in Puppies’ Food Marketed in Bulk in Southern Brazil

Background: The Brazilian domestic canine population are the second largest in the world and their feeding means 0.4% of the Brazilian gross domestic product. For maintaining the quality of the food, the companies use worldwide standards for technical prevention and control of contaminants and biological conservation. The packaging is part of this process, since it provides a barrier between food and environment. However, in Brazil, packagings are often opened in retail stores for bulk marketing. The objectives of this work were to develop a methodology to detect viruses in foods and to analyze the bacterial and fungal contamination in puppies’ food sold in bulk in Ivoti and Estância Velha, cities in Southern Brazil. Materials &amp; Methods &amp; Results: Twenty samples collected between September and October 2016 were analyzed for most probable number of coliforms, Salmonella sp., mesophilic aerobic bacteria and yeast/mold following the regulation of the Brazilian Ministry of Agriculture, Livestock and Food Supply guidelines. They were also tested for Human Mastadenovirus C (HAdV), Canine Mastadenovirus A (CAdV), and Carnivore Protoparvovirus 1 (CPV) genomes. Viral analysis were performed by polymerase chain reaction (PCR) detection. During the collection of the samples hygienic-sanitary conditions, storage of feeds, animals’ access, dog grooming, and veterinary care were considered to evaluate the conditions of each store. A pilot study was carried out using one food sample marketed in bulk and one sample from the original package (closed package) and testing them for bacterial and fungal contamination for standardizing viral detection. Ten grams of food from the original package were mixed with 90 mL of Eagles’ Minimal Essential Medium (E-MEM) in 100 mL sterile bottles. These bottles were kept in room temperature and shaken for 60 min. Subsequently, aliquots were obtained by sequentially diluting the sample (10-2 to 10-4). All final specimens contained 10 mL and each diluted sample was spiked with HAdV-C prototype viral strain (AdV5). A standard solution of HAdV-C was diluted from 3.6x106TCID50mL, (50% tissue culture infective dose) to 3.6x103 TCID50mL, and DNA extraction was performed. Nested-PCR targeting AdV DNApol was performed to detect adenoviruses from different hosts. AdV-positive samples were submitted to nucleotide sequencing and phylogenetic analysis. Specific PCRs were also carried out for CAdV and CPV. Mesophilic aerobic bacteria were detected in all samples and Aspergillus sp. was found in five samples, among which one sample was co-infected with Penicillium sp. One sample was positive for AdV, which was identified as HAdV by sequencing; while coliforms and Salmonella sp. were not detected.Discussion: The presence of fungi with mycotoxigenic potential, such as Aspergillus sp. and Penicillium sp. represents a threat for canines, due to toxins that may persist for a long period even after the fungus is not viable. Moreover, dogs seem to be more susceptible to the effects of the toxins, which is probably because of low glutathione s-transferase activity. Some species of Penicillium sp. genus may produce ocrathoxin A, which nephrotoxic and immunosuppressive effects in dogs are widely reported. Mesophilic bacteria were detected in all samples (at 104 CFU/g) and considered harmless. The detection of human viruses points to the presence of anthropic contamination; on the other hand, ingestion of contaminated feed, even if it is by a heterologous species, turns the dogs into carriers of the virus. In addition, manipulation of those feeds by children who share the same space with dogs can result in gastroenteritis episodes.

Biochemical and molecular biomarkers in integument biopsies of free-ranging coastal bottlenose dolphins from southern Brazil

Adverse effects of exposure to persistent organic pollutants (POPs) threaten the maintenance of odontocete populations. In southern Brazil, coastal bottlenose dolphins from the Laguna Estuarine System (LES) and Patos Lagoon Estuary (PLE) were sampled using remote biopsies during the winter and summer months. Levels of bioaccumulated POPs were measured in the blubber. The activities of glutathione S-transferase (GST), glutathione reductase (GR), glutathione peroxidase (GPx), and superoxide dismutase (SOD) were also quantified, as were the mRNA transcript levels of aryl hydrocarbon receptor (AhR), AhR nuclear translocator (ARNT), cytochrome P450 1A1-like (CYP1A1), metallothionein 2A (MT2A), GST-π, GPx-4, GR, interleukin 1 alpha (IL-1α), and major histocompatibility complex II (MHCII) in the skin. In general, levels of POPs were similar among sites, sexes, ages and seasons. For most animals, total polychlorinated biphenyl (ΣPCBs) levels were above the threshold level have physiological effects and pose risks to cetaceans. The best-fitting generalized linear models (GLMs) found significant associations between GR, IL-1α and GPx-4 transcript levels, SOD and GST activities, and total polybrominated diphenyl ether (ΣPBDEs) and pesticide levels. GLMs and Kruskal-Wallis analyses also indicated that there were higher transcript levels for most genes and lower GST activity in the winter. These results reinforce the need to consider the influence of environmental traits on biomarker values in wildlife assessments.

High serum iron and zinc decrease glutathione S-transferase among women with breast cancer

BACKGROUNDBreast cancer in Indonesia ranks second as the most common cancer found in women after cervical cancer. Tumor cells express increased levels of antioxidant proteins to detoxify reactive oxygen species (ROS). Glutathione S-transferase (GST) is known to play a key role in the detoxification and reduction of ROS. Trace elements have nutritional benefits as essential cofactors for physiologic processes, but some can be toxic to humans. Accumulated evidence suggests that deficiency or excess of certain trace elements may be associated with risk of chronic diseases including cardiovascular diseases, diabetes and cancer. The aim of this research was to determine the correlation of the trace elements iron and zinc with the oxidative stress level of GST activity in women with breast cancer.METHODSAn observational analytic study with cross sectional design was conducted involving 35 breast cancer women. Research subjects were women with breast cancer diagnosed by biopsy. Measurements of iron and zinc levels were performed using atomic absorption spectrometry, GST activity was measured using spectrophotometry. Data analysis was done with the Pearson correlation test.RESULTSThe results showed that there was a significant relationship of serum iron with GST (r=-0.487; p&lt;0.05) and serum zinc with GST (r=-0.409; p&lt;0.01).CONCLUSIONOur study demonstrated that the higher zinc and iron level, the lower GST level among breast cancer women. It is recommended to use zinc, iron and GST levels as biomarkers for breast cancer and its progression.

The activity of erythrocyte and platelet glutathione reductase and glutathione-S-transferase in paranoid schizophrenia

A comparative evaluation of glutathione reductase (GR) and glutathione-S-transferase (GST) activities in erythrocytes and platelets of patients with schizophrenia. Fifty patients, 47 men and 3 women, aged 25-56 years (medium 34) with acute paranoid schizophrenia (F20.0 ICD-10) with hallucinatory-paranoid or paranoid syndrome were studied. The control group consisted of 48 healthy people, 45 men and 3 women, aged 21-59 years (medium 38). GR activity was determined by the oxidation of NADP-H in the reduction reaction of oxidized glutathione. GST activity was determined by the rate of chromogenic conjugate formation between glutathione and 1-chloro-2.4-dinitrobenzene. No differences in the erythrocyte GR and GST activities between the control group and patients with schizophrenia were found. The platelet activity of GR and GST was significantly lower in patients compared to the control group (Mann-Whitney U test, p<0.01). Spearman rank correlation analysis showed that the erythrocyte GST activity was significantly correlated with PANSSneg scores when measured at the beginning of the study, GST was higher in those patients who had less PANSSneg scores after treatment (R=-0.41, p<0.05). The activity of platelet GST in patients with schizophrenia was correlated with the severity of positive symptoms (PANSSpos score) at the beginning of the study before taking therapy (R=-0.31, p<0.05), i.e. the more prominent psychotic symptoms were expressed in patients with lower GST activity. Upon completion of therapy, this association disappeared. The activity of glutathione-dependent enzymes in the blood cells of schizophrenic patients determined before the beginning of antipsychotic pharmacotherapy may be important for objective assessment of this metabolic system status and the degree of its impairment in patients.

Efficient Two-Photon Fluorescent Probe for Glutathione S-Transferase Detection and Imaging in Drug-Induced Liver Injury Sample.

Drug-induced liver injury (DILI) is a potential complication of any prescribed medication. So far, the diagnosis of DILI is still a clinical challenge due to the lack of efficient diagnosis method. Glutathione S-transferase (GST), with a high concentration in liver cytosol, can reduce toxicity and facilitate urinary excretion by catalyzing the conjugation of glutathione (GSH) with reactive metabolites in liver. When liver is seriously damaged, GST and GSH will be released into plasma from liver cytosol, which caused a lower GST activity in liver cytosol. Therefore, monitoring the level of GST activity in liver tissue may be a potential strategy for diagnosis of DILI. Here, we reported a two-photon probe P-GST for GST activity detection for the first time. In the proposed design, a donor-π-acceptor (D-π-A) structured naphthalimide derivative with efficient two-photon properties was chosen as the fluorescent group, and a 2,4-dinitrobenzenesulfonate group was employed as the GST recognition unit, which also acted as the fluorescence quencher. In the present of GST and GSH, the recognition unit was removed and the fluorophore was released, causing a 40-fold enhancement of fluorescence signal with a detection limit of 35 ng/mL. At last, P-GST was successfully applied in two-photon imaging of GST in cells and DILI samples, which demonstrated its practical application in complex biosystems as a potential method for diagnosis of DILI.

Retraction Note to: Coordinated response of sulfate transport, cysteine biosynthesis, and glutathione-mediated antioxidant defense in lentil (Lens culinaris Medik.) genotypes exposed to arsenic.

Response of sulfate transporters, thiol metabolism, and antioxidant defense system was studied in roots of two lentil (Lens culinaris Medik.) genotypes grown in arsenic (10, 25, and 40 μM As(V))-supplemented nutrient solution, and significant changes compared to control (0 μM As(V)) were observed mainly at 25 and 40 μM. In L 414, high glutathione (GSH) redox (0.8-0.9) was maintained with elevated thiol synthesis, powered by transcriptional up-regulation of LcSultr1;1 and LcSultr1;2 sulfate transporters and significant induction of LcSAT1;1 and LcSAT1;2 (serine acetyltransferase), OAS-TL (O-acetylserine(thiol)-lyase), γ-ECS (γ-glutamylcysteine synthetase), and PCS (phytochelatin synthase) genes predominantly within 12-24 h of As exposure at 25 μM and within 6-12 h at 40 μM. This thiolic potency in L 414 roots was effectively complemented by up-regulation of gene expressions and consequent enhanced activities of superoxide dismutase, ascorbate peroxidase (APX), dehydroascorbate reductase, glutathione reductase (GR), and glutathione-S-transferase (GST) isoforms at 25 and 40 μMAs, efficiently scavenging excess reactive oxygen species to prevent onset of As-induced oxidative stress and consequent inhibition of root growth in L 414. In contrast, down-regulation of vital sulfate-uptake transporters as well as entire thiol-metabolizing system and considerably low APX, GST, and GR expressions in DPL 59 not only resulted in reduced GSH redox but also led to over-accumulation of H2O2. This triggered membrane lipid peroxidations as the marks of As-induced oxidative damage. Results indicated coordinated response of thiol-metabolism and antioxidant defense in conferring As-tolerance in lentil, and GSH is the key point in this cascade.

Variations in hepatic biomarkers in American alligators (Alligator mississippiensis) from three sites in Florida, USA

Sub-individual biomarkers are sub-lethal biological responses commonly used in the assessment of wildlife exposure to environmental contaminants. In this study, we examined the activity of glutathione-s-transferase (GST) and lactate dehydrogenase (LDH), and metallothionein (MT) concentrations among captive-raised alligator hatchlings, wild-caught juveniles, and wild-caught adults. Juveniles and adults were collected from three locations in Florida (USA) with varying degrees of contamination (i.e. Lake Apopka (organochlorine polluted site), Merritt Island National Wildlife Refuge (NWR) (metal polluted site), and Lake Woodruff NWR (reference site)). We examined whether changes in the response of these three biomarkers were age and sex dependent or reflected site-specific variations of environmental contaminants. Juvenile alligators from Merritt Island NWR had higher MT concentrations and lower GST activity compared to those from the other two sites. This outcome was consistent with higher metal pollution at this location. Sexually dimorphic patterns of MT and GST (F > M) were observed in juvenile alligators from all sites, although this pattern was not observed in adults. GST activity was lower in captive-raised alligators from Lake Apopka and Merritt Island NWR as compared to animals from Lake Woodruff NWR, suggesting a possible developmental modulator at these sites. No clear patterns were observed in LDH activity. We concluded that GST and MT demonstrate age and sex specific patterns in the alligators inhabiting these study sites and that the observed variation among sites could be due to differences in contaminant exposure.

Differential metabolism of 4-hydroxynonenal in liver, lung and brain of mice and rats

The lipid peroxidation end-product 4-hydroxynonenal (4-HNE) is generated in tissues during oxidative stress. As a reactive aldehyde, it forms Michael adducts with nucleophiles, a process that disrupts cellular functioning. Liver, lung and brain are highly sensitive to xenobiotic-induced oxidative stress and readily generate 4-HNE. In the present studies, we compared 4-HNE metabolism in these tissues, a process that protects against tissue injury. 4-HNE was degraded slowly in total homogenates and S9 fractions of mouse liver, lung and brain. In liver, but not lung or brain, NAD(P)+ and NAD(P)H markedly stimulated 4-HNE metabolism. Similar results were observed in rat S9 fractions from these tissues. In liver, lung and brain S9 fractions, 4-HNE formed protein adducts. When NADH was used to stimulate 4-HNE metabolism, the formation of protein adducts was suppressed in liver, but not lung or brain. In both mouse and rat tissues, 4-HNE was also metabolized by glutathione S-transferases. The greatest activity was noted in livers of mice and in lungs of rats; relatively low glutathione S-transferase activity was detected in brain. In mouse hepatocytes, 4-HNE was rapidly taken up and metabolized. Simultaneously, 4-HNE-protein adducts were formed, suggesting that 4-HNE metabolism in intact cells does not prevent protein modifications. These data demonstrate that, in contrast to liver, lung and brain have a limited capacity to metabolize 4-HNE. The persistence of 4-HNE in these tissues may increase the likelihood of tissue injury during oxidative stress.

A key role for an impaired detoxification mechanism in the etiology and severity of autism spectrum disorders.

BackgroundAutism Spectrum Disorders (ASD) is a syndrome with a number of etiologies and different mechanisms that lead to abnormal development. The identification of autism biomarkers in patients with different degrees of clinical presentation (i.e., mild, moderate and severe) will give greater insight into the pathogenesis of this disease and will enable effective early diagnostic strategies and treatments for this disorder.MethodsIn this study, the concentration of two toxic heavy metals, lead (Pb) and mercury (Hg), were measured in red blood cells, while glutathione-s-transferase (GST) and vitamin E, as enzymatic and non-enzymatic antioxidants, respectively, were measured in the plasma of subgroups of autistic patients with different Social Responsiveness Scale (SRS) and Childhood Autism Rating Scale (CARS) scores. The results were compared to age- and gender-matched healthy controls.ResultsThe obtained data showed that the patients with autism spectrum disorder had significantly higher Pb and Hg levels and lower GST activity and vitamin E concentrations compared with the controls. The levels of heavy metals (Hg and Pb), GST and vitamin E were correlated with the severity of the social and cognitive impairment measures (SRS and CARS). Receiver Operating Characteristics (ROC) analysis and predictiveness curves indicated that the four parameters show satisfactory sensitivity, very high specificity and excellent predictiveness. Multiple regression analyses confirmed that higher levels of Hg and Pb, together with lower levels of GST and vitamin E, can be used to predict social and cognitive impairment in patients with autism spectrum disorders.ConclusionThis study confirms earlier studies that implicate toxic metal accumulation as a consequence of impaired detoxification in autism and provides insight into the etiological mechanism of autism.