Useful Marker Of Oxidative Stress Research Articles

Low intensity whole-body oxidative stress in patients with parotid gland tumors.

Recently, there has been a significant increase in the number of patients hospitalized and undergoing surgery due to parotid gland tumors. We have observed an increase in the total number of salivary gland tumors, and hence an increased number of malignant tumors. Etiological factors of salivary gland tumors are still unknown. Oxidative stress (OS) has been suggested to be an important indicator of carcinogenesis in some tumors. Therefore, the aim of our study was to assess the state of OS in patients with selected benign and malignant parotid gland tumors. After final postoperative histological diagnosis of the tumor, patients were divided into the following groups i.e. pleomorphic adenoma 35 patients, Warthins tumor (WT) - 17 patients, mucoepidermoid carcinoma - 9 patients and 45 controls. The final study group included 106 patients. We found an increase in plasma lipofuscin levels in patients with all investigated parotid gland tumors. Furthermore, the activity of Cu/Zn-SOD in Warthins tumor patients was statistically lower compared to the controls, pleomorphic adenoma or mucoepidermoid carcinoma.In conclusion, we sustain the hypothesis on the presence of a low intensity whole-body OS in patients with parotid gland tumors. Furthermore, LPS may be a useful marker of OS in patients with benign and malignant tumors. Moreover, pathological mechanisms connected with WT seem to be different compared to malignant neoplasms and other benign tumors. It seems that Cu/Zn-SOD, as an antioxidative enzyme, may play an important role in those mechanisms taking part in WT.

Activity of the enzyme delta-aminolevulinate dehydratase and parameters of oxidative stress in different modes of delivery

Aims The objective of this study was to investigate the effect of the type of delivery (vaginal and cesarean) on the activity of the enzyme delta-aminolevulinate dehydratase (δ-ALA-D), which as yet has not been studied, and the markers of oxidative stress. Methods Seventy-six mothers and their newborns were divided into two groups: normal birth (VD) and elective cesarean section (ECS). Samples of maternal and umbilical cord blood were collected up to 5 min after birth. Thiobarbituric acid reactive substances (TBARS), advanced oxidation protein products (AOPP), protein thiol (P-SH), nonprotein (NP-SH), the ferric reducing ability of plasma (FRAP), total antioxidant capacity (TAC), catalase, and δ-ALA-D enzyme activity were tested. Results TBARS and AOPP were significantly higher in mothers of the VD group, while P-SH, NP-SH, FRAP and TAC were reduced. In newborns, TBARS and AOPP did not differ between the groups; however, in the VD group, there was a decrease in P-SH, NP-SH, FRAP, TAC, and catalase. The activity of the δ-ALA-D enzyme was decreased in mothers and neonates born by VD. Conclusions Mothers undergoing VD had higher levels of free radicals and lower antioxidant defenses, while their newborns decreased antioxidant defenses likely to contain the oxidative imbalance. The inhibition of the δ-ALA-D enzyme in this scenario allows its use as a useful marker of oxidative stress in different obstetric settings.

Serum enzymes markers in myocardial infarction: A study of rural area

Background: Acute Myocardial Infarction is the reduction of coronary flow to such an extent that supply of oxygen to the myocardium do not need the oxygen demand of myocardial tissues. The diagnosis of AMI cannot be fulfilled unless the elevated levels of serum cardiac enzymes particularly CK-MB iso-enzyme activity. The rate of release of cardiac enzymes is highly diagnostic.Aims and Objectives: Therefore, the present thesis aim is-in evaluating, whether the elevated levels of cardiac marker enzymes can be compared to the extent of the Myocardial infarction.Material and Methods: Therefore the present study was undertaken on 50 patients of MI and 50 patients of control. They were group according to the age and sex. The activity of different cardiac enzymes were studied.Result: The AMI patients had significantly elevated levels of 90% patients of high LDH values and 86% patients have elevated levels of AST. The significant elevation of serum enzymes as compared to the control.Conclusion: The magnitude of the elevated levels of enzymes can be compared to the extent of the myocardial infarction. Serum GGT can also be useful marker of oxidative stress in myocardial infarction.Asian Journal of Medical Sciences Vol.8(2) 2017 34-37

Plasma marker of tissue oxidative damage and edaravone as a scavenger drug against peroxyl radicals and peroxynitrite.

The percentage of the plasma oxidized form of coenzyme Q10 in the total amount of coenzyme Q10 (%CoQ10) is a useful marker of oxidative stress in the circulation. Plasma free fatty acids and their composition can be used as markers of tissue oxidative damage, as demonstrated in patients suffering from a wide variety of diseases and in humans and rats under oxidative stress. Edaravone was approved for the treatment of stroke in Japan in 2001 and its mechanism of action is based on scavenging lipid peroxyl radicals. In 2015, edaravone was also approved for the treatment of ALS patients. Edaravone functions therapeutically as a scavenger of peroxynitrite, as demonstrated by the finding that its administration raises plasma uric acid levels and decreases 3-nitrotyrosine in cerebrospinal fluid.

Oxidative Stress in Vivax Malaria

Malaria is still a leading cause of morbidity and mortality. The increase in lipid peroxidation reported in malaria infection and antioxidant status may be a useful marker of oxidative stress during malaria infection. The aim of this study was to investigate the role of antioxidant enzymes against toxic reactive oxygen species in patients infected with Plasmodium vivax and healthy controls. Malondialdehyde levels, superoxide dismutase, and glutathione peroxidase activities were determined in 91 P. vivax patients and compared with 52 controls. Malondialdehyde levels, superoxide dismutase, and glutathione peroxidase activities were 8.07±2.29 nM/ml, 2.69±0.33 U/ml, and 49.6±3.2 U/g Hb in the patient group and 2.72±0.50 nM/ml, 3.71±0.47 U/ml, and 62.3±4.3 U/g Hb in the control group, respectively. Malondialdehyde levels were found statistically significant in patients with vivax malaria higher than in healthy controls (P<0.001). On the other hand, superoxide dismutase and glutathione peroxidase activities were found to be significantly lower in vivax malaria patients than in controls (P<0.05). There was an increase in oxidative stress in vivax malaria. The results suggested that antioxidant defense mechanisms may play an important role in the pathogenesis of P. vivax.

Urinary Catalytic Iron in Obesity

Obesity precedes the development of many cardiovascular disease risk factors, including type 2 diabetes mellitus (DM), hypertension, and chronic kidney disease. Catalytic iron, which has been associated with these chronic diseases, may be one of the links between obesity and these multifactorial diverse disorders. We investigated whether urinary catalytic iron is increased in obese individuals without DM and overt kidney disease. We measured urinary catalytic iron using established methods in 200 randomly selected individuals without DM [100 who were obese (body mass index ≥30 kg/m(2)) and 100 who were nonobese (body mass index ≤27)]. Participants were selected from an outpatient clinic and community setting and were part of an ongoing cross-sectional study of obesity in individuals between the ages of 18 and 70 years. There was a significant difference in mean (95% CI) urinary catalytic iron excretion between the obese participants and the nonobese participants, 463 (343-582) nmol/mg [52.3 (38.8-65.8) nmol/μmol] vs 197 (141-253) nmol/mg [22.3 (15.9-28.6) nmol/μmol]; P < 0.001. The significant predictors of increased urinary catalytic iron were obesity (P = 0.001) and waist-to-hip ratio (P = 0.03). Our study results demonstrate that obesity and waist-to-hip ratio are associated with increased urinary catalytic iron, which may be a useful marker of oxidative stress. Additional studies are needed to determine the role of catalytic iron in increased cardiovascular disease and chronic kidney disease associated with obesity.

Antioxidant effects of a dietary supplement: Reduction of indices of oxidative stress in normal subjects by water-soluble chitosan

The effect of water-soluble chitosan, a natural polymer derived from chitin, on indices of oxidative stress was investigated in normal volunteers. Treatment with chitosan for 4 weeks produced a significant decrease in levels of plasma glucose, atherogenic index and led to increase in high density lipoprotein cholesterol (HDL). Chitosan treatment also lowered the ratio of oxidized to reduced albumin and increased total plasma antioxidant activity (TPA). There was good correlation between TPA and oxidized albumin ratio. The results indicate that oxidized albumin ratio represents a potentially useful marker of oxidative stress. In in vitro studies, albumin carbonyls and hydroperoxides were significantly decreased in a time-dependent manner in the presence of chitosan, compared with controls ( p < 0.05). Chitosan also reduced two stable radicals in a dose- and time-dependent manner. The results suggest that chitosan has a direct antioxidant activity in systemic circulation by lowering the indices of oxidative stress in both in vitro and in vivo studies. This may confer benefits additional to the reduction in plasma carbohydrate and increase in HDL levels. It may also inhibit oxidation of serum albumin commonly observed in patients undergoing hemodialysis, resulting in reduction of oxidative stress associated with uremia.

Electronegative LDL and Lipid Abnormalities in Patients Undergoing Hemodialysis and Peritoneal Dialysis

Background: Oxidative modification of low-density lipoprotein (LDL) has been demonstrated in patients with end-stage renal disease, where it is associated with oxidative stress and plays a key role in the pathogenesis of atherosclerosis. In this context, the generation of minimally oxidized LDL, also called electronegative LDL [LDL(–)], has been associated with active disease, and is a detectable sign of atherogenic tendencies. The purpose of this study was to evaluate serum LDL(–) levels and anti-LDL(–) IgG autoantibodies in end-stage renal disease patients on dialysis, comparing patients on hemodialysis (HD), peritoneal dialysis (PD) and a control group. In addition, the serum lipid profile, nutritional status, biochemical data and parameters of mineral metabolism were also evaluated. Methods: The serum levels of LDL(–) and anti-LDL(–) IgG autoantibodies were measured in 25 patients undergoing HD and 11 patients undergoing PD at the Centro Integrado de Nefrologia, Rio de Janeiro, Brazil. Ten healthy subjects served as a control group. Serum levels of albumin, total cholesterol, triglycerides and lipoproteins were measured. Calculations of subjects’ body mass index and measurements of waist circumference, triceps skin fold and arm muscle area were performed. Measurements of hematocrit, serum blood urea nitrogen, creatinine, parathyroid hormone, phosphorus and calcium were taken. Results: Levels of LDL(–) were higher in HD patients (575.6 ± 233.1 µg/ml) as compared to PD patients (223.4 ± 117.5 µg/ml, p < 0.05), which in turn were higher than in the control group (54.9 ± 33.3 µg/ml, p < 0.01). The anti-LDL(–) IgG autoantibodies were increased in controls (0.36 ± 0.09 µg/ml) as compared to PD (0.28 ± 0.12 µg/ml, p < 0.001) and HD patients (0.2 ± 0.1 µg/ml, p < 0.001). The mean values of total cholesterol and LDL were considered high in the PD group, whereas the mean triceps skin fold was significantly lower in the HD group. Conclusion: Levels of LDL(–) are higher in renal patients on dialysis than in normal individuals, and are reciprocally related to IgG autoantibodies. LDL(–) may be a useful marker of oxidative stress, and this study suggests that HD patients are more susceptible to cardiovascular risk due to this condition. Moreover, autoantibodies reactive to LDL(–) may have protective effects in chronic kidney disease.

Succination of Protein Thiols during Adipocyte Maturation

Although obesity is a risk factor for development of type 2 diabetes and chemical modification of proteins by advanced glycoxidation and lipoxidation end products is implicated in the development of diabetic complications, little is known about the chemical modification of proteins in adipocytes or adipose tissue. In this study we show that S-(2-succinyl)cysteine (2SC), the product of chemical modification of proteins by the Krebs cycle intermediate, fumarate, is significantly increased during maturation of 3T3-L1 fibroblasts to adipocytes. Fumarate concentration increased > or =5-fold during adipogenesis in medium containing 30 mm glucose, producing a > or =10-fold increase in 2SC-proteins in adipocytes compared with undifferentiated fibroblasts grown in the same high glucose medium. The elevated glucose concentration in the medium during adipocyte maturation correlated with the increase in 2SC, whereas the concentration of the advanced glycoxidation and lipoxidation end products, N(epsilon)-(carboxymethyl)lysine and N(epsilon)-(carboxyethyl)lysine, was unchanged under these conditions. Adipocyte proteins were separated by one- and two-dimensional electrophoresis and approximately 60 2SC-proteins were detected using an anti-2SC polyclonal antibody. Several of the prominent and well resolved proteins were identified by matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry. These include cytoskeletal proteins, enzymes, heat shock and chaperone proteins, regulatory proteins, and a fatty acid-binding protein. We propose that the increase in fumarate and 2SC is the result of mitochondrial stress in the adipocyte during adipogenesis and that 2SC may be a useful biomarker of mitochondrial stress in obesity, insulin resistance, and diabetes.

Up-regulation of hMUTYH, a DNA repair enzyme, in the mitochondria of substantia nigra in Parkinson’s disease

There is ample evidence for the involvement of oxidative stress in mitochondrial DNA damage and repair mechanisms in Parkinson's disease (PD). The human MutY homolog (hMUTYH) which removes misincorporated adenine opposite 8-oxoG in DNA functions in post-replication, and is localized in the nuclei and mitochondria. We hypothesized that hMUTYH is involved in the disease process of PD. To test our hypothesis, we performed immunohistochemical and biochemical studies on brains of patients with PD and those of control patients. Our results showed up-regulation of hMUTYH in the mitochondria of the SN of PD patients. Western blot analysis also revealed high hMUTYH levels in PD patients and expression of a 47-kDa molecule in the brains as the major isoform. This molecule was localized within the mitochondria as confirmed by double staining with a mitochondrial marker. To confirm the presence of this molecule, we examined the mRNAs of isoforms that translate to the 47-kDa molecule. Based on the amount of mRNAs, the major molecule was alpha4. Interestingly, this molecule lacks the mitochondria targeting sequence. Our results suggest that hMUTYH might be a useful marker of oxidative stress and that oxidative stress and genomic instability are important in the PD disease process.

Lipid peroxidation in type 2 diabetes mellitus

AIM: The link between hyperglycemia, enhanced freeradical activity, and the complications of diabetes is unknown. The purpose of this study is to evaluate the levels of malondialdehyde (MDA) measured as thiobarbituric acid-reactive substances (TBARS), an index of lipid peroxidation, in patients of type 2 diabetes without complications, and compare with normal subjects of the same population. METHODOLOGY: We recruited 60 type 2 diabetic subjects without complications and with poor metabolic control and 60 age-matched controls with good metabolic control. Levels of glucose, total cholesterol, HbA 1C , and MDA as TBARS were determined. RESULTS: Diabetic patients had higher levels of blood glucose (P<0.001), HbA 1C (P<0.001), and MDA (P<0.001) than control subjects. The total cholesterol of the control subjects and diabetic patients did not differ. There was no correlation between the family history in diabetics and elevation in either HbA or 1C MDA levels. CONCLUSION: Increased levels of MDA may be a useful marker of oxidative stress. The enhanced lipid peroxidation leads to an increase in free-radical activity in type 2 diabetics. This increase in free-radical activity in type 2 diabetes mellitus along with insulin resistance can lead to activation of stress-sensitive pathways, which may play an important role in the complications of diabetes.

Glutathione regulates susceptibility to oxidant-induced mitochondrial DNA damage in human lymphocytes

Oxidative damage to mitochondrial DNA (mtDNA) interferes with the expression of mitochondrial-encoded subunits of the electron transport complexes of oxidative phosphorylation. MtDNA is protected by several mitochondrial antioxidant systems, but the specific importance of glutathione is unknown. We hypothesized that glutathione protects mtDNA from oxidative damage in human blood lymphocytes and that glutathione depletion increases susceptibility to mtDNA depletion, which increases vulnerability to apoptosis. MtDNA damage was measured in human blood lymphocytes exposed to tert-butyl-hydroperoxide (t-BOOH) or t-BOOH plus the glutathione analog, glutathione ethyl ester (GEE). Mitochondrial oxidative stress, mtDNA damage, and susceptibility to apoptosis were analyzed after glutathione depletion with buthionine sulfoximine (BSO). The data show selective damage to lymphocyte mtDNA at low concentrations of tBOOH that is attenuated by glutathione supplementation. Moreover, inhibition of glutathione synthesis led to lymphocyte ROS generation and mtDNA damage, and increased susceptibility to receptor-mediated apoptosis. These findings implicate the glutathione system in maintaining mtDNA integrity and resistance to apoptosis in lymphocytes and suggest that assessment of mtDNA damage in blood lymphocytes may be a useful marker of oxidative stress in humans.

Selective quantification of free 3-nitrotyrosine in exhaled breath condensate in asthma using gas chromatography/tandem mass spectrometry

Reactive nitrogen species can cause oxidative modifications of certain amino acid residues in proteins, notably the modification of tyrosine to 3-nitrotyrosine (3-NT), which is a potentially useful marker of oxidative stress. Since lung diseases are associated with airway inflammation and oxidative stress, quantification of 3-NT in exhaled breath condensate (EBC) may provide a non-invasive means for monitoring ongoing inflammatory processes. 3-NT-like immunoreactivity has previously been detected in EBC, but no definitive evidence for the presence of 3-NT in EBC is available. Here, a method based on gas chromatography/negative ion chemical ionization/tandem mass spectrometry was established for the quantification of free 3-NT in EBC. The detection limit was 0.56 pM (corresponding to 3.0 amol microl(-1) sample injected) and the method was found to give linear results (r2 > 0.999) in the concentration range of 0-5.0 nM. The coefficient of variation (CV) for within-day and between-day precision were 11 and 12%, respectively. No artifactual nitration was observed during sample processing. The method was applied to study subjects with asthma (n = 8), and healthy subjects (n = 10), but only a slight non-significant increase in 3-NT levels was found in the former group (median [interquartile ranges]; 99 [50-547] amol s(-1) vs. 75 [35-147] amol s(-1)). No correlation with exhaled nitric oxide (NO), pulmonary function or EBC levels of total protein was observed. The 3-NT levels were much lower compared to previously reported levels, based on immunochemical measurements. The method does not allow the simultaneous quantification of tyrosine in samples.

Urinary 20-hydroxyeicosatetraenoic acid excretion is associated with oxidative stress in hypertensive subjects

Oxidative stress has been implicated in the pathogenesis of hypertension. 20-Hydroxyeicosatetraenoic acid (20-HETE) is a cytochrome P450 product of arachidonic acid metabolism, thought to be involved in the regulation of blood pressure (BP). The metabolism of arachidonic acid by cytochrome P450 enzymes may be a significant source of oxidative stress. F 2-isoprostanes are reliable markers of in vivo oxidative damage. γ-Glutamyl transpeptidase (γ-GT) has traditionally been associated with alcohol intake or liver dysfunction and may be an early marker of oxidative stress. The objective of the present study was to investigate relationships between 20-HETE excretion and markers of oxidative stress (F 2-isoprostanes and γ-GT). Sixty-nine treated hypertensive subjects underwent measurement of 24-h ambulatory BP, serum γ-GT, and urinary F 2-isoprostane and 20-HETE excretion. 20-HETE excretion was positively associated with 24-h diastolic BP ( p = 0.005), alcohol intake ( p = 0.008), γ-GT ( p = 0.007), and F 2-isoprostanes ( p = 0.005). F 2-isoprostanes were positively associated with alcohol intake ( p = 0.018) and γ-GT ( p = 0.01). In a multivariate regression, γ-GT remained an independent predictor of 20-HETE excretion, after adjustment for age, gender, BMI, and alcohol intake. In conclusion, the study highlights the positive association observed between 20-HETE excretion and markers of oxidative damage. The study also provides evidence that γ-GT may be a useful marker of oxidative stress.

Accumulation ofNε-(Hexanoyl)lysine, an Oxidative Stress Biomarker, in Rice Seeds during Storage

Nepsilon-(hexanoyl)lysine (HEL) is a potentially useful marker of oxidative stress in animals. We investigated whether HEL might be useful as a marker in rice seeds damaged by oxidative stress during storage, as well as in animals. The germination ability of rice decreased with lipid peroxidation during storage at 40 degrees C for three months. Moreover, we observed accumulation of HEL in the damaged rice. In addition, the activities of antioxidative enzymes, catalase and superoxide dismutase, significantly decreased in the rice seeds during storage at 40 degrees C. These results suggest that HEL might be a useful marker of oxidative stress in rice.

Induced Sputum 8-Isoprostane Concentrations in Inflammatory Airway Diseases

Induced sputum 8-iso-prostaglandin F(2alpha) (PGF(2alpha)) concentrations may be a useful marker of oxidative stress in airways disease. This study examines oxidative stress (measured by 8-iso-PGF(2alpha)) in airway disease according to disease type (asthma and bronchiectasis), disease activity (stable and acute asthma), and disease pattern (intermittent, mild, moderate, and severe persistent asthma). We compared subjects with stable asthma (n = 71) and bronchiectasis (n = 23) with healthy control subjects (n = 29). Another group of patients with asthma (n = 39) were assessed during and after acute exacerbation. Induced sputum 8-iso-PGF(2alpha) concentrations were validated and found to be elevated in subjects with stable asthma and bronchiectasis versus control subjects (median [interquartile range] 216 [103-389] and 698 [264-1,613] ng/L vs. 123 [41-290] ng/L, p < 0.001) and increased as clinical asthma pattern worsened (intermittent 115 [42-153], mild persistent 116 [89-229] ng/L, moderate persistent 183 [110-317] ng/L, severe persistent 387 [102-587] ng/L; p = 0.010). Sputum 8-iso-PGF(2alpha) concentrations were elevated during acute asthma and decreased with recovery (458 [227-950] ng/L vs. 214 [148-304] ng/L, p = 0.0002). We conclude that 8-iso-PGF(2alpha) is involved in the pathophysiology of inflammatory airway diseases, being related to disease type, pattern, and activity. Analysis of 8-iso-PGF(2alpha) concentrations in induced sputum provides a useful tool for monitoring oxidative stress and investigating strategies aimed at reducing oxidative stress in airways disease.

Oxidative stress in patients with non-complicated malaria

Aim: To compare oxidative stress in adults with non-complicated malaria and healthy controls. Methodology: We measured malondialdehyde (MDA), total antioxidant status (TAS), catalase, superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX). Oxidative stress was calculated based on MDA/TAS, MDA/GSH-PX and SOD/catalase indexes. Results: Mean MDA in patients was 3.9 μmol/L (controls = 1.3 μmol/L). Mean TAS was 0.9 mmol/L in patients and controls. Malaria patients had less catalase activity when compared to controls (209.4 vs. 320.4 k/gr), while SOD and GSH-PX activity was higher (79.4 U/mL, 11,884.2U/L vs. 54.3 U/mL, 9,672.6 U/L). MDA/TAS index was 3.5 fold more in patients than in controls, MDA/GSH-PX and SOD/catalase indexes were increased by 6 and 2.8 fold. MDA levels and MDA/TAS index showed no differences according to malarial history, parasitaemia, Plasmodium species, parasite’s stage, place of residence and drinking or smoking habits. Conclusions: During acute non-complicated P. falciparum or P. vivax malaria, we observed high oxidative stress. This resulted from lipid peroxidation rather than from a reduced TAS. We propose MDA/TAS index as a useful marker of oxidative stress during malaria infection.

Protein carbonyl group content in patients affected by familiar chronic nail candidiasis.

Familiar chronic nail candidiasis (FCNC) is a rare disorder characterized by early-onset infections caused by different species of Candida, restricted to the nail of the hands and feet, and associated with a low serum concentration of intercellular adhesion molecule 1. Host defense mechanisms against candidiasis require the cooperation of many immune cells through several candidacidal mechanisms, including oxygen-dependent killing mechanisms, mediated by a superoxide anion radical myeloperoxidase--H2O2--halide system, and reactive nitrogen intermediates. We analyzed protein carbonyl groups (considered a useful marker of oxidative stress) in the serum of patients belonging to a five-generation Italian family with an isolated form of FCNC. Serum protein carbonyl groups in FCNC patients were significantly lower than those measured in healthy donors. Also, if this hypothesis is merely speculative, we could suggest that the decreased circulating level of protein carbonyl groups in these patients is not a marker of a lower oxidative stress condition, but might be linked to a lower protease activity.

Characterization of a Novel Hemoglobin-Glutathione Adduct That Is Elevated in Diabetic Patients

Typically, a diagnosis of diabetes mellitus is based on elevated circulating blood glucose levels. In an attempt to discover additional markers for the disease and predictors of prognosis, we undertook the characterization of HbA1d3 in diabetic and normal patients. PolyCAT A cation exchange chromatography and liquid chromatography-mass spectroscopy was utilized to separate the alpha- and beta-globin chains of HbA1d3 and characterize their presence in normal and diabetic patients. We report the characterization of HbA1d3 as a glutathionylated, minor hemoglobin subfraction that occurs in higher levels in diabetic patients (2.26 +/- 0.29%) than in normal individuals (1.21 +/- 0.14%, p < 0.001). The alpha-chain spectrum displayed a molecular ion of m/z 15126 Da, which is consistent with the predicted native mass of the HbA0 alpha-globin chain. By contrast, the mass spectrum of the beta-chain showed a mass excess of 307 Da (m/z = 16173 Da) versus that of the native HbA0 beta-globin chain (m/z = 15866 Da). The native molecular weight of the modified beta-globin chain HbA0 was regenerated by treatment of HbA1d3 with dithiothreitol, consistent with a glutathionylated adduct. We propose that HbA1d3 (HbSSG) forms normally in vivo, and may provide a useful marker of oxidative stress in diabetes mellitus and potentially other pathologic situations.

Oxidative DNA damage, antioxidants and DNA repair: applications of the comet assay

Estimates of background levels of oxidative base damage in human white blood cells vary enormously, from 300 down to 0.4 molecules of 8-oxoguanine per 106 guanines. An EC-funded Concerted Action, the European Standards Committee on Oxidative DNA Damage, is currently attempting to resolve the discrepancy and to agree a realistic estimate of basal endogenous oxidation. Oxidation of lymphocyte DNA is a useful marker of oxidative stress, and this can be decreased by supplementation with pure antioxidants or with foods rich in antioxidants. The steady-state level of DNA oxidation is ultimately controlled by the process of DNA repair; the extent to which this varies between individuals has yet to be established.