Glutathione sulfur transferases (GSTs) is a group of enzymes involved in the detoxification process of carcinogens and other substances. The genes encoding isoenzymes M1 and T1 are polymorphic in humans and the phenotypic absence of enzyme activity (null genotype) may have an effect on the risk of chronic lymphoblastic leukemia (CLL). Our purpose was to examine whether the GSTM1 and GSTT1 homozygous null genotypes altered the risk of CLL. DNA was extracted from the peripheral blood of 27 patients with CLL and 147 cancer-free controls; both groups originated from a defined population (residents of the Ioannina region, northwestern Greece) and were similar with regard to mean age, race and sex; GSTM1 and GSTT1 were simultaneously analyzed by a multiplex polymerase chain reaction (PCR) method and Fisher's exact test was used for comparisons between the two groups. A significantly increased incidence of the GSTM1 null genotype was found in the group of patients compared to the controls (74.07 versus 34.69%, P = 0.0002). Additionally, the incidence of the GSTT1 null genotype was comparable in patients and controls (25.92 versus 10.20%, P = 0.05). The frequency of the combined GSTM1 null/T1 null genotype was significantly different between patients and controls (14.81 versus 2.04%, P = 0.012). However, there was no relationship between the advanced stage of the disease and the GSTs status. Individuals with the GSTM1 and GSTT1 null genotypes may have enhanced susceptibility to CLL.

Oxidative stress and accumulation of free radicals might play a role in the pathogenesis of vitiligo. Glutathione S-transferase (GST) is a multigene family of enzymes that detoxify oxidative stress products. In this study, genotyping by multiplex PCR of GSTM1 and GSTT1 in 101 women with nonsegmental vitiligo vulgaris and 101 age-matched healthy female volunteers showed that only the GSTM1 null genotype (P=0.04) was significantly overexpressed in patients with vitiligo. Analysis of the combined effect of GSTM1 and GSTT1 genotyping identified a significant association of risk for vitiligo with the GSTT1/GSTM1 double-null type only (P=0.01; OR=2.69; 95% CI 1.12-6.46). Age of onset of vitiligo was significantly earlier in patients with the T1 null genotype (P<0.01) and those with the T1-/M1+ and T1-/M1- combined genotypes (P<0.01 and P=0.01, respectively). In conclusion, the GSTM1 gene and the GSTM1/GSTT1 double-null genotype may be a risk factor for vitiligo in Egyptian patients. Inability to cope with oxidative stresses because of GST deficiency may cause early disease onset.

Several studies have shown that high oxidative stress levels are associated with varicocele. The GST (glutathione S-transferase) family of genes is critical in the protection of cells from oxidative stress because they utilise as substrates a wide variety of products of oxidative stress. The objective of this study was to assess the relationship between genetic polymorphism in GST-M1 and GST-T1 and varicocele using 109 varicocele patients and 123 controls. Varicoceles were clinically graded as Grade 1, Grade 2 and Grade 3. GST-M1 and GST-T1 genes were determined by polymerase chain reaction. Although the GST-M1 null genotype was higher in Grade 3 than in Grade 1, 2 and controls, there were no statistical differences between control group and varicocele groups according to GST-M1 and GST-T1 null genotype. Men with varicocele do not have more GST-M1 and GST-T1 null polymorphisms than men without varicocele. Additional studies are needed to assess the exact mechanism by which the varicocele corresponds to elevated ROS levels.

OBJECTIVE

The aim of this study was to examine chromosome aberrations in 25 tunnel workers exposed to acrylamide-containing grout in injection work.

METHODS

Blood samples were collected from 25 exposed and 25 unexposed tunnel workers matched for age, gender, and smoking habits. Whole blood was cultured for 50-53 hours according to conventional methods. Chromosome damage was scored in 200 metaphases per person on coded slides. The distribution of glutathione S-transferase (GST) genotypes (M1 and T1) was examined for all the workers. Exposure assessment was performed with detailed interviews and questionnaires.

RESULTS

The chromosome examinations showed no statistically significant differences between the 25 exposed and 25 unexposed workers for cells with chromosome aberrations or for chromatid breaks, chromosome breaks, and chromosome gaps. The exposed workers had a significantly higher number of chromatid gaps (mean 10.6, SD 5.6) than the unexposed workers (mean 6.4, SD 4.4, P=0.004), but there was no exposure-response relationship. The limited stratum-specific numbers showed that the exposed workers with the GSTM1-/GSTT1-genotype had nonsignificantly higher frequencies of all the effect parameters than the unexposed workers; this finding indicates that individual susceptibility related to the detoxification of acrylamide and N-methylolacrylamide may have played a role in the observed effect.

CONCLUSIONS

No increase in chromosome breaks or aberrations was observed for 25 workers exposed to acrylamide-containing grout during tunnel work. The increased frequency of chromatid gaps in the exposed workers may indicate a slight genotoxic effect related to exposure to acrylamide or N-methylolacrylamide.

OBJECTIVE

Inherited impairment of xenobiotic metabolism is a postulated mechanism underlying environmentally associated pathogeneses such as multiple chemical sensitivity (MCS). Using the Quick Environmental Exposure and Sensitivity Inventory (QEESI), we defined people who have a strong response to chemical substances as "chemical sensitive populations (CSP)." The aim of this study is to evaluate the condition of subjects sensitive to chemicals and to analyze their genotypes in order to identify susceptibility factors in CSPs in Japanese populations.

METHODS

A total of 1,084 employees of Japanese companies were surveyed using the QEESI, history of MCS, and sick house syndrome. The common genotypes of the participants were analyzed for glutathione S-transferase (GST) M1, GSTT1, aldehyde dehydrogenase2 (ALDH2), and paraoxonase1 (PON1) in order to identify factors in the susceptibility to sensitivity to chemicals.

RESULTS

Four subjects had history of diagnosis of MCS; no subjects had diagnosis of sick house syndrome. The subjects were divided into four levels according to scores of 0, 1-19, 20-39, and 40 or more on three of the QEESI subscales. In addition, we used the MCS criteria by Hojo to differentiate between cases (CSP) and controls. No significant differences in the allelic distribution of genetic polymorphisms in the GSTM1, GSTT1, ALDH2 or PON1 genes were found among the four levels of each subscale, or between cases and controls.

CONCLUSIONS

Our findings suggest that the common genotypes of GSTM1, GSTT1, ALDH2, and PON1 are of little importance to CSP in a Japanese population.

Preeclampsia is a pregnancy-specific disorder in humans and a major cause of maternal and neonatal morbidity and mortality. Increasing evidence suggests that oxidative stress plays an important role in the pathogenesis of preeclampsia. The aim of this study was to investigate the relationship between null alleles of the glutathione S-transferases (GST) M1 and T1 genes and the risk of preeclampsia. This case-control study involved 112 preeclamptic and 233 normoevolutive pregnant women. The null polymorphisms were genotyped by multiplex polymerase chain reaction (PCR). Our results showed an increased risk of preeclampsia in patients with the GSTT1 null genotype [odds ratio (OR) = 2.21; 95% confidence interval (CI) = 1.14-4.27; P = 0.018]. Our data further showed that a combination of deletion genotypes of the GSTM1 and GSTT1 genes conferred an even higher risk of preeclampsia (OR = 4.56, 95%CI = 1.59-13.09; P = 0.005). Our results provide the first evidence suggesting that a GSTT1 null polymorphism might be associated with preeclampsia in the Mexican mestizo population, and that this risk increases with the combination of both GSTT1 and GSTM1 null polymorphisms.

The frequency of sister chromatid exchanges (SCEs), high SCE frequency cells (HFCs), and genetic polymorphism of genotypes glutathione S-transferase (GST) M1 and T1 were analyzed in peripheral lymphocytes of 35 workers occupationally exposed to chromium (Cr) and 35 matched control group. Results showed that workers exposed to Cr showed 6.07 SCE/cell, as compared to 4.76 SCE/cell for the control group (p<0.01). Smokers showed a statistically significant higher frequency of SCE than non-smokers in both groups. The work duration of Cr workers was an important factor. Workers exposed for more than 5 years showed a significantly higher level of SCEs (p<0.05). Workers exposed to Cr for 5 or more years had higher HFC rates (51.4%) than those exposed for less than 5 years (22.9%), with an odds ratio of 4.5 times than those exposed for less than 5 years. In HFC analysis, Cr workers who smoked showed a higher level of HFC (60%) than the control group (5.7%) and also had a higher odds ratio (60.4) compared with the control group. Among non-smokers, the odds ratio was 9.0. Another objective of this study is to investigate the relationship between SCE and genetic polymorphisms of GST M1 and T1 in Cr workers. The results showed that the incidence of GSTM1 null genotype was 60% in the control group and 77.1% in Cr workers, and percentages of GSTT1 deletion were 42.9% and 62.9% in control and exposed individuals, respectively. There was a slightly increased frequency of SCE among Cr workers with GSTM1 null genotype as opposed to non-null genotype individuals. A similar result was seen among the control group; however, there were no statistically significant differences. In conclusion, the current study found the positive induction of SCE in workers who smoked or/and were exposed to Cr. However, different GST genotypes did not influence the level of cytogenetic damage between groups. Despite slight variation in numbers, they all appear to be not different.

OBJECTIVE

Polymorphisms of glutathione S-transferase (GST) genes in mothers may be involved in teratogenesis in their offspring. This study aims to investigate the association of GST genes (T1, M1 and P1) with the risk of having children with congenital malformations (CMs) in residents of the West Siberian region of Russia.

METHODS

We studied 235 women with offspring's with CMs, and 273 women with one or more healthy children. Null genotypes of GSTM1 and GSTT1 were identified through multiplex real-time polymerase chain reaction, and GSTP1 gene (Ile105Val) polymorphism was determined through TaqMan-real-time polymerase chain reaction.

RESULTS

The study showed that the maternal genotype GSTT1 «0/0» is associated with CMs in the offspring (odd ratio (OR) = 3.63, P = 5.18 × 10(-9) ). A significant association of the maternal genotype GSTT1 «0/0» with CMs of the cardiovascular system (OR = 5.03, P = 2.93 × 10(-7) ), urinary system (OR = 4.20, P = 3.51 × 10(-6) ) and central nervous system (OR = 4.40, P = 6.69 × 10(-5) ) was found in the child. No association of maternal GSTM1 (del) and GSTP1 (Ile105Val) genetic polymorphisms with CMs of the child was identified.

CONCLUSIONS

Homozygous deletion of the GSTT1 gene in women of the West Siberian region is a risk factor for birth defects in the child.

1. The metabolism of a nitrate ester-substituted dihydropyridine derivative (NND) in vitro was characterized with rabbit hepatic microsomes and cytosol. 2. Denitration activity was located in both the microsomal and cytosolic fractions, whereas oxidation to the pyridine analogue was solely located in the microsomal fraction. 3. Oxidation to the pyridine analogue required NADPH and was inhibited by carbon monoxide, miconazole and SKF-525A, suggesting that oxidation was catalysed by P450. 4. Denitration activity in the microsomes required either NADPH or GSH. Together with these results, responses to various inhibitors indicate participation of both P450 and glutathione S-transferase (GST). 5. Denitration activity in cytosol was activated by glutathione (GSH), and by dithiothreitol (DTT) to a greater extent. GSH-dependent denitration was inhibited by S-hexyl GSH, an inhibitor of GST, but DTT-dependent denitration was not. Moreover, the formation patterns of the mono-denitrated metabolites, M1 and M2, were shown to be different in each incubation condition. 6. These results suggest that the denitration of NND in cytosol could be catalysed by a GSH-independent enzyme as well as the GSH-dependent enzyme, GST.

The objectives of this study were to investigate the individual characteristics, lifestyle habits, exposure levels, and genetic diversity of xenobiotic-metabolizing enzymes involved in toluene metabolism in Korean and foreign workers exposed to toluene at a manufacturing plant. This study was conducted to determine the effects of culture or ethnicity on toluene metabolism. The results showed that blood and urinary toluene concentrations were dependent on the level of exposure to toluene. We analyzed the correlation between toluene metabolism and genetic diversity in glutathione S-transferase (GST) (M1), GSTT1, and cytochrome p-450 (CYP) 2E1*5 as well as lifestyle habits (smoking, drinking, and exercise habits). The results revealed significant correlations between toluene metabolism and GSTM1 and GSTT1 genetic diversity, as well as smoking and exercise.

Glutathione transferase (GST) enzymes catalyze the conjugation of glutathione with reactive functional groups of endogenous compounds and xenobiotics, including halonitroaromatics. 1-Chloro-2,4-dinitrobenzene (CDNB) is one of the most commonly used substrates for GST activity assays. We have studied the interactions of dinitronaphthalene analogues of CDNB with recombinant human GST enzymes (Alpha, Mu, and Pi classes) expressed in Escherichia coli. Dinitronaphthalene derivatives were found to be GST inhibitors. The highest potency of inhibition was observed towards Mu-class GSTs, M1-1 and M2-2; IC50 values for 1-methoxy- and 1-ethoxy-2,4-dinitronaphthalene were in the high nanomolar to low micromolar range. Inhibition accompanies the formation, at the enzyme active site, of very stable Meisenheimer complex intermediates.

Glutathione S-transferase (GST) family of enzymes are involved in a two-stage detoxification process of a wide range of environmental toxins, carcinogens and xenobiotics. The GST enzymes play important roles in oxidative stress pathways, and polymorphisms in the GSTM1 and GSTT1 genes mediate susceptibility and outcome in different diseases. Human immunodeficiency virus (HIV) infection is associated with oxidative stress, but there is limited data on the frequency of deleted GSTM1 and GSTT1 genes in HIV/AIDS patients and their effect on progression among Ghanaians. This study sought to investigate the association between homozygous deletion of GSTM1 and GSTT1 genes (both null deletion) with HIV/AIDS disease progression in Ghanaian patients. HIV-infected individuals on antiretroviral therapy (ART), ART-naïve HIV patients, and HIV seronegative individuals were recruited for the study. HIV/AIDS disease progression was assessed by measuring CD4+ cell count and viral load of the patients, and GST polymorphism was determined by amplifying the GSTT1 and GSTM1 genes using multiplex PCR, with CYP1A1 gene as an internal control. The mean CD4+ count of patients that were naïve to ART (298 ± 243 cells/mm3) was significantly lower than that of patients on ART (604 ± 294 cells/mm3), and viral load was significantly lower in the ART-experienced group (30379 ± 15073 copies/mm3) compared to the ART-naïve group (209882 ± 75045 copies/mm3). Frequencies of GSTM1 and GSTT1 deletions were shown to be 21.9% and 19.8%, respectively, in the HIV patients, and patients with homozygous deletion of both GSTM1 and GSTT1 were more likely to have their CD4+ count rising above 350 cells/mm3 (OR = 6.44, 95% CI = 0.81-51.49, p = 0.039) suggesting that patients with homozygous deletion of GSTM1 and GSTT1 genes have slower disease progression. The findings of this study show that double deletion of glutathione S-transferases M1 and T1 is statistically associated with normal CD4+ count in patients diagnosed with HIV/AIDS. Further study is required to investigate the clinical importance of the both null deletion in HIV patients.

Human exposure assessments for perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS) have been mostly limited to the quantification of these chemicals in different environmental matrices, but only a few studies have addressed toxicological aspects associated with them. It has been suggested that both PFOA and PFOS are highly stable chemicals that are not metabolized, yet previous reports have described abnormal activity of important biotransformation pathways. Therefore, the goal of the present study was to investigate the effects of PFOA and PFOS on phase I and II biotransformation enzymes at the gene expression and activity levels, and by using the well-established human liver HepaRG cell line. Cells were exposed to a wide range of PFOA and PFOS concentrations for 24 or 48 h, prior to cytotoxicity measurements, and quantification of expression and activity of three cytochrome P450 enzymes (CYP1A2, CYP2C19 and CYP3A4) and two conjugation enzymes (glutathione-S-transferase (GST-M1) and UDP-glucuronosyltransferase (UGT-1A1)). Expression of all CYP enzymes was significantly reduced from exposure to both PFOA and PFOS after 48 h and from concentrations as low as 40-50 ng/L, with CYP3A4 also presenting the lowest activity. Among the conjugation enzymes, the expression of UGT was significantly reduced only by PFOA after 48 h of exposure, yet no significant alterations in its activity were observed. While the specific chemico-biological interactions of these compounds with gene expression and biotransformation pathways is not clear, the results from this study suggest that the interference of PFOA and PFOS with phase I and II biotransformation enzymes could potentially lead to adverse outcomes resulting from the inability of biotransformation pathways to function as needed.

OBJECTIVE

To explore the relationship between DNA damage induced by vinyl chloride monomer (VCM) and polymorphisms of metabolizing enzymes of VCM.

METHODS

Comet assay was employed to detect DNA damage, the workers were divided into two groups based on the status of DNA damage. Case-control design was used to investigate the relationship between the genetic polymorphisms of metabolizing enzymes and DNA damage. Genotypes of CYP2E1 c1/c2 and mEH4 His139Arg were identified by the PCR-RFLP, null genotypes of GST T1 and GST M1 were detected by PCR.

RESULTS

The genotypes of CYP2E1 c1c2 and c2c2 were significantly associated with DNA damages (P < 0.01). A prominent risk increasing of DNA damage was observed for those individuals having a high or low VCM exposure and possessing the CYP2E1 c1c2 and c2c2 genotypes (OR 4.92, 95% CI 1.35-13.85 and OR 2.57, 95% CI 1.01-6.59).

CONCLUSIONS

Cumulative exposure dose and polymorphism of metabolizing enzymes may modulate the DNA damage of VCM-exposed workers. possessing the CYP2E1 c1c2 and c2c2 genotypes (OR 4.92, 95% CI 1.35-13.85 and OR 2.57, 95% CI 1.01-6.59).

CONCLUSIONS

Cumulative exposure dose and polymorphism of metabolizing enzymes may modulate the DNA damage of VCM-exposed workers.

OBJECTIVE

The influence of the polymorphic human glutathione-S-transferase (GST) T1 and M1 genotypes (classified as "conjugators" and "nonconjugators") on the biological effects of nonoccupational ethylene oxide exposure as reflected by the formation of globin N2-hydroxyethylvaline adducts was investigated. Specific attention was paid to smoking as a potential source of exposure. A total of 27 Caucasian subjects, including 10 women and 17 men, participated in the study. Volunteers were grouped as smokers, i.e., 6 subjects (5 male, 1 female), and nonsmokers, i.e., 21 subjects (12 male, 9 female). The regular cigarette consumption in the smoker group ranged from 10 to 25 cigarettes/day.

METHODS

The amount of N2-hydroxyethylvaline (HEV) bound to the N-terminal valine in human globin was determined following a procedure described by Bader and co-workers and the Deutsche Forschungsgemeinschaft. The GST genotypes were determined by a polymerase chain reaction (PCR) analysis outlined by Bell and colleagues (with beta-globin serving as an internal standard) and Kempkes and co-workers (coamplification of the GSTT1 fragment).

RESULTS

The median level of HEV detected in the smoker group was 280 pmol/g globin as compared with the median value of 50 pmol/g globin recorded for the nonsmokers, indicating that ethylene oxide intake from cigarettes may result in a approximately 5-fold higher overall HEV level in smokers in comparison with nonsmoking individuals. No dose-effect correlation was observed between daily cigarette consumption and the resulting HEV levels. Moreover, the individual GSTT1 or GSTM1 genotype did not influence the HEV level in smokers. The subgroup of nonsmoking GSTT1 conjugators revealed a median HEV value of 46 pmol/g globin as compared with the median value of 92 pmol/g globin found in the nonconjugator subgroup. Subjects with the GSTM1 gene had a median HEV value of 55 pmol/g globin, whereas subjects with the gene deletion had a median HEV value of 44 pmol/g globin. The 2-fold increase in the median HEV value detected in GSTT1 non-conjugators as compared with GSTTI conjugators indicates an influence of GSTT1 on the globin HEV levels. This hypothesis was found to be significant in the nonparametric Mann-Whitney U-test for independent samples (P < 0.002, two-sided). The same test was applied to evaluate the influence of GSTM1 on the globin HEV levels. No significant influence was observed (P>> 0.10; two-sided).

CONCLUSIONS

This result is in accordance with the finding that ethylene oxide is a substrate for GSTT1 but not for GSTM1. In addition, this study demonstrates a clear influence of genetically determined GSTT1 status on biological effects, e.g., protein adduct formation after non-occupational ethylene oxide exposure. In smokers, however, a modulating influence of GSTT1 status was not observed.

The biotransformation of the dietary carcinogen aflatoxin B1 (AFB1) was examined in hepatic microsomal and cytosolic fractions from channel catfish, an aquatic species shown to be refractory to AFB1 toxicity and reported to be resistant to AFB1 hepatocarcinogenesis. Catfish liver microsomes catalyzed the in vitro oxidation of AFB1 to the reactive AFB1-8,9-epoxide (AFBO) at high substrate concentrations (128 microM AFB1) but not at low substrate concentrations (16 microM AFB1) which were more representative of environmental exposure. A similar trend was observed for the production of the hydroxylated metabolite aflatoxin M1 (AFM1). In contrast, hepatic microsomes prepared from rainbow trout, a species sensitive to AFB1 toxicity and hepatocarcinogenesis, activated AFB1 at 16 and 128 microM AFB1, with the rate of AFB1 epoxidation by trout microsomes exceeding that of catfish by more than sixfold. Treatment of channel catfish with 5,6-benzoflavone (beta NF, 20 mg/kg) resulted in a threefold increase in AFM1 formation but did not affect AFBO formation. AFB1 was rapidly reduced to aflatoxicol (AFL), a putative detoxification product of AFB1, at both low and high substrate concentrations. The rate of AFL production by channel catfish hepatic cytosol was 40- and 65-fold greater than observed for rainbow trout at 16 and 128 microM AFB1, respectively. Western blotting of catfish cytosols revealed the presence of a catfish cytosolic protein of approximately 25 kDa that displayed immunological cross-reactivity to rat GST Ya, but not to other rat or mouse alpha class GSTs with high AFBO-conjugating activity. Furthermore, catfish cytosolic GSTs did not catalyze the conjugation of AFBO with GSH. The results of these studies indicate that AFB1 is poorly oxidized by channel catfish microsomes, and suggest that the lack of microsomal AFB1 activation together with the rapid conversion of AFB1 to AFL contributes to the apparent resistance of channel catfish to AFB1 toxicity and hepatocarcinogenesis.

Tobacco use and environmental air pollution are the established etiological factors in head and neck cancer (HNC) progression. Nevertheless, not all the inhabitants with high usage of tobacco from the same polluted locality are suffering with HNC and this is due to the existence of factors like inter-individual genetic polymorphisms, life time exposure to tobacco and the rate of xenobiotic metabolism enzyme (XME) activity. The present study investigates the polymorphic genotypes of the most important XME, glutathione-S-transferase Mu 1 (GST M1) and Theta 1 (GST T1) as the risk modulator to HNC among tobacco-habituated inhabitants of Saurashtra in Gujarat, a region in western India. A population based case–control study was done in 252 HNC patients and 504 healthy controls. Blood samples were collected from the subjects and investigated for polymorphic genotypes of GST M1 and GST T1. Estimation of the odds of risks was done by logistic regressions. Among the subjects with high usage of tobacco, M1 not null-T1 null genotypes presence was found as risk reducing factor to HNC with 0.334 folds (95 % CI; 0.170-0.659). The presence of M1 null-T1 not null genotypes was found with susceptibility to HNC among the subjects with no habit of tobacco chewing, adjusted odds ratio (AOR) 3.170 (1.128-8.913) and no habit of smoking, AOR of 2.544 (1.094-5.963). The present study reveals the finding of significantly increased risk to HNC by interactions of GST M1 null-GST T1 not null polymorphic genotypes among the subjects with nil or less tobacco usage shed some light for the insights of biomarker application in early detection of HNC.

BACKGROUND

Tobacco contains agents which generate various potent DNA adducts that can cause gene mutations. Production of DNA adducts may be neutralized by glutathione S transferase (GST) along with other phase I and phase II enzyme systems. The existence of null type of GST among the population increases the susceptibility to various disorders and diseases. The present study focuses on the impact of high tobacco usage and possible null type mutation in GST loci.

METHODS

Genotypes of GST were detected by multiplex polymerase chain reaction in unrelated 504 volunteers of high tobacco using natives of Gujarat. Allelic frequencies were calculated using Statistical Package for Social Studies-16 software. Hardy Weinberg Equilibrium (HWE) was calculated using Chi square test. Two sided Fisher's significance test was used to compare allelic frequencies of different populations.

RESULTS

The frequency of homozygous null genotype of GSTM1 and GSTT1 were 20% (95% CI 16.7-23.9) and 35.5% (95% CI 31.4-39.9) respectively. The GSTM1 and GSTT1 null allele frequency distribution in the Gujarat population was significantly deviating from HWE. GSTT1 null frequency of Gujaratians was significantly higher and different to all reported low tobacco using Indian ethnics, while GSTM1 was not differing significantly.

CONCLUSIONS

Tobacco usage significantly influences the rate of mutation and frequency of GSTT1 and M1 null types among the habituates. The rate of mutation in GSTT1 loci was an undeviating response to the dose of tobacco usage among the population. This mutational impact of tobacco on GSTT1 postulates the possible gene - environment interaction and selection of null genotype among the subjects to prone them under susceptible status for various cancers and even worst to cure the population with GSTT1 dependent drugs.

BACKGROUND

Cardiomyocytes are particularly susceptible to complications from iron loading. The blood transfusions in thalassaemia major create loading of iron that cannot be naturally excreted. Apolipoprotein E and Glutathione S-transferase act as the scavenger of free radicals, which are generated due to excess iron. The variants of Apolipoprotein E (ApoE) and Glutathione S-transferase (GST) may play a role in oxidative damage-induced cardiomyopathy, so we aimed to study the association of genetic variants of these genes on diastolic dysfunction in our patients.

METHODS

One hundred and five β-thalassaemia patients older than 10 years were enrolled for the study. Two-dimensional and M-mode echocardiography analysis was done in all patients. Genotyping of the genetic variants of aforementioned genes was done using the PCR-RFLP method. Serum Glutathione S-transferase levels were estimated by ELISA.

RESULTS

Diastolic dysfunction was observed in 24 (22.8%) patients, whereas left ventricular hypertrophy was present in 37(35.2%) patients. There was a significant association of GSTM1 null allele with diastolic dysfunction only. Serum GST levels were also positively correlated with e/a and e/e' ratio. Positive association of ApoE E2 allele with the diastolic dysfunction was also seen.

CONCLUSIONS

Patients having Glutathione S-transferase M1 allele and Apolipoprotein E E2 allele are predisposed to oxidative stress-induced cardiac injury.

Glutathione S-transferases, including GST-T1 and GST-M1, are known to be involved in the phase II detoxification pathways for xenobiotics as well as in the metabolism of endogenous compounds. Polymorphisms in these genes have been linked to an increased susceptibility to carcinogenesis and associated with risk factors that predispose to certain inflammatory diseases. In addition, GST-T1 and GST-M1 null genotypes have been shown to be responsible for interindividual variations in the metabolism of arsenic, a known human carcinogen. To assess the specific GST genotypes in the Mexican population chronically exposed to arsenic, we have developed a multiplex High Resolution Melting PCR (HRM-PCR) analysis using a LightCycler480 instrument. This method is based on analysis of the PCR product melting curve that discriminates PCR products according to their lengths and base sequences. Three pairs of primers that specifically recognize GST-T1, GST-M1, and β-globin, an internal control, to produce amplicons of different length were designed and combined with LightCycler480 High Resolution Melting Master Mix containing ResoLight, a completely saturating DNA dye. Data collected from melting curve analysis were evaluated using LightCycler480 software to determine specific melting temperatures of individual melting curves representing target genes. Using this newly developed multiplex HRM-PCR analysis, we evaluated GST-T1 and GST-M1 genotypes in 504 DNA samples isolated from the blood of individuals residing in Zimapan, Lagunera, and Chihuahua regions in Mexico. We found that the Zimapan and Lagunera populations have similar GST-T1 and GST-M1 genotype frequencies which differ from those of the Chihuahua population. In addition, 14 individuals have been identified as carriers of the double null genotype, i.e., null genotypes in both GST-T1 and GST-M1 genes. Although this procedure does not distinguish between biallelic (+/+) and monoallelic (+/-) genotypes, it can be used in an automated workflow as a simple, sensitive, and time and money saving procedure for rapid identification of the GST-T1 and GST-M1 positive or null genotypes.

BACKGROUND

Null genotypes of glutathione S-transferase (GST) exhibit the absence of enzymatic activity and are associated with increased cardiovascular risk. Recent reports have related both lower and higher urinary sodium excretion (USE) to higher cardiovascular risk. Here we investigate the impact of GSTM1 and GSTT1-null polymorphisms on USE in a Brazilian population.

METHODS

We cross-sectionally evaluated 1,308 subjects from the city of Vitoria, Brazil, based on clinical history, physical examination, anthropometry, analysis of laboratory parameters, measurement of USE, and GST polymorphisms genotyping.

RESULTS

The frequency of GST M1, T1, and double-deletion polymorphisms was 51%, 22%, and 11%, respectively. Individuals with the GSTM1-null genotype had lower USE than those with the non-null genotype (92.1±52.3 vs. 102.8 ± 6 0.7 mEq/12h; P < 0.001). Linear regression analysis adjusted for confounding factors revealed that the GSTM1-null genotype was independently associated with USE (P = 0.001). In addition, diastolic blood pressure and triglyceride levels were higher in GSTM1-null individuals than in non-null individuals in the highest tertile of USE. Finally, the presence of GSTT1-null or double-deleted genotypes did not influence USE or affect the interactions between USE and the variables studied.

CONCLUSIONS

Deletion of GSTM1 was associated with low USE and modulated the interaction between sodium intake and blood pressure in Brazilian subjects. These novel findings may provide a new unexplored link between sodium regulation and GST homeostasis.

OBJECTIVE

To explore the relationship between genetic polymorphisms of glutathione S-transferase (GST) M1, T1 and susceptibility to mountain sickness.

METHODS

Forty-three soldiers with acute mountain sickness and 80 healthy soldiers matching with sex/age and training under the same condition were divided into case group and control group. A multiple polymerase chain reaction method was used to detect GSTM1 and GSTT1 genes in genomic DNA isolated from peripheral blood cells from both cases and controls.

RESULTS

The frequency of the GSTT1 positive genotype was significantly higher in cases (69.8%) than in controls (42.5%) (P = 0.004, OR = 3.12, 95% CI 1.42 approximately 6.86). The frequency of GSTM1 negative genotype was also higher in cases (72.1%) than in controls (52.5%) (P = 0.03, OR = 2.34, 95% CI 1.05 approximately 5.02). Persons with both GSTM1 and GSTT1 negative genotypes had 5-fold more risk than those with GSTT1 negative and GSTM1 positive genotypes in developing mountain sickness (OR = 5.04, 95% CI: 1.00 approximately 25.3).

CONCLUSIONS

Genetic polymorphisms of glutathione S-transferase M1, T1 may be the risk factors in the development of mountain sickness.

The expression of different isoenzymes of glutathione transferase (GST), i.e. the cytosolic subunits GSTA1/A2, A3, A4, A5, M1/2, M2 and P1, T2, and the microsomal GST in follicles of different sizes and in corpora lutea from porcine ovary, was investigated by Western blotting. No immunoreactivity was obtained with anti-rat GSTT2 or anti-rat microsomal GST polyclonal antibodies. In contrast, GSTA1/A2, A3, A4, A5, M1/2, M2 and P1 are all expressed in the cytosol from porcine ovaries. In general, the highest levels of these GST isoenzymes were present in the cytosol from corpora lutea, in agreement with measurements of activity towards 1-chloro-2,4-dinitrobenzene. Immunoreactivity with anti-rat GSTP1 was only obtained with follicles. The cytosolic GSTs from follicles and corpora lutea were affinity purified on glutathione-Sepharose and separated by reversed-phase high-performance liquid chromatography in order to quantitate the different subunits. A peak corresponding to the class pi subunit was present in follicles. This peak was also seen with corpora lutea, although at very low level. There were four peaks containing class mu subunits. The remaining peaks were concluded to contain the class alpha subunits, except for two peaks which are suggested to contain proteins other than GSTs. The levels of the different subunits were quantitated on the basis of the areas under the peaks and the relative amounts in follicles of different sizes and in corpora lutea corresponded well with the Western blot analysis.