Erythrocyte Cu-Zn Superoxide Dismutase Research Articles

Non-genomic, direct modulatory effect of 17β-estradiol, progesterone and their synthetic derivatives on the activity of human erythrocyte CuZn superoxide dismutase

This study aimed to evaluate whether natural or synthetic steroid hormones could directly modulate the activity of the different superoxide dismutase (SOD) isoforms found in human blood fractions without changing enzyme expression. Enzyme samples of human erythrocytes, the human platelet-rich plasma fraction (PRP) or isolated CuZnSOD, which was purified from human erythrocytes were pre-incubated with natural steroids (17β-estradiol 17-acetate and progesterone) and their synthetic derivatives (β-estradiol 3-benzoate and medroxyprogesterone 17-acetate). Then, CuZn and MnSOD activities were measured using the xanthine/xanthine oxidase/nitroblue tetrazolium method. Hormones had no effect on MnSOD activity from the PRP, but we show for the first time that natural and synthetic steroid hormones have a direct, bell-shaped effect on the activity of CuZnSOD from both male and female human erythrocytes. Low (physiological) hormone concentrations caused a dose-dependent increase in enzyme activity, which disappeared at higher hormone concentrations. In addition, the combination of synthetic and natural estrogens and progestins had a synergistic stimulatory effect on the activity of CuZnSOD from human erythrocytes. The molecular interaction between CuZnSOD and steroid hormones was preliminarily studied. Natural hormones did not change the electrophoretic mobility of SOD under denaturing conditions, but they did increase the absorption spectra of SOD in the 230–290 nm range. These data suggest that hormone-mediated modulation of CuZnSOD is related to subtle changes in protein conformation, possibly related to Trp and Phe residues. We propose that this effect may account for the physiological regulation of enzyme activity during conditions where steroid hormones undergo alterations as the ovulatory cycle.

Nitric oxide and superoxide dismutase in rheumatoid arthritis: Correlation with disease activity

Aim: To analyse serum and synovial fluid nitric oxide (NO) concentrations and erythrocyte CuZn superoxide dismutase (SOD) activity in rheumatoid arthritis (RA) patients and to determine the importance of imflammatory markers in reflect disease activity. Methods: 20 RA patients and 15 healthy controls were included. NO level was determined spectrophotometrically by the Griess reaction and CuZn SOD activity by the method of Sun et al. Results: Erythrocyte sedimentation rate (ESR), C-reactive protein and leukocyte levels were determined as inflammatory markers. Significantly higher ESR (p<0.001) and leukocyte levels (p<0.01) and serum and synovial fluid NO levels (p<0.01, p<0.05) and lower CuZn SOD activity (p<0.05) were observed in RA patients. Classification of RA patients according to disease activity score (DAS) revealed significantly higher synovial fluid NO level (p<0.05) and lower CuZn SOD activity (p<0.05) in RA patients with DAS≥2.7. Conclusions: Increased serum and synovial fluid NO levels and decreased CuZn SOD activity up-regulated inflammatory response are observed in RA patients.

Flexibility of the Cu,Zn superoxide dismutase structure investigated at 0.57 GPa

The 2 A resolution crystal structure of bovine erythrocyte Cu,Zn superoxide dismutase (CuZnSOD) has been determined by X-ray diffraction at high pressure (0.57 GPa) and room temperature. At 0.57 GPa the secondary, tertiary and quaternary structures are similar to other previously determined bovine erythrocyte CuZnSOD structures. Nevertheless, pressure has a localized impact on the atomic coordinates of C(alpha) atoms and on side chains. The compression of the crystal and of the protein backbone is anisotropic. This anisotropy is discussed, taking into account intermolecular contacts and protein conformation. Pressure perturbation highlights the more flexible zones in the protein such as the electrostatic loop. At 0.57 GPa, a global shift of the dimetallic sites in both subunits and changes in the oxidation state of Cu were observed. The flexibility of the electrostatic loop may be useful for the interaction of different metal carriers in the copper-uptake process, whereas the flexibility of the metal sites involved in the activity of the protein could contribute to explaining the ubiquitous character of CuZnSODs, which are found in organisms living in very different conditions, including the deep-sea environment. This work illustrates the potential of combining X-ray crystallography with high pressure to promote and stabilize higher energy conformational substates.

Erythrocyte Indices, Microminerals and Ratios, Antioxidants and Lipids in Centrum Materna Diet-Supplemented Omani Mothers

Venous (maternal) and cord blood (neonatal) samples of Omani women who had a daily supplement of Centrum Materna multivitamin and multimineral tablet throughout pregnancy were investigated at late preterm (n=37) and at term (n=37) delivery for erythrocyte indices, micromineral, antioxidant, and lipid values. Hemoglobin (Hb), hematocrit (HCT), mean cell volume (MCV), red cell distribution width (RDW), copper (Cu), zinc (Zn), ceruloplasmin, erythrocyte Cu-Zn superoxide dismutase (Cu-Zn SOD), cholesterol, apolipoprotein (apo) A-I and apo B were measured by appropriate analytical systems. Cu/zinc and Cu/ceruloplasmin ratios were calculated. The erythrocyte indices were normal in neonatal blood but showed borderline anemia in maternal blood of both groups. There were significantly decreased values of Cu (P=0.012), Zn (P=0.001), apo A-I (P=0.029), and Cu/ceruloplasmin ratio (P=0.032) in late preterm compared to term mothers. Significantly decreased values of Cu (P=0.003), ceruloplasmin (P<0.0001), apo A-I (P=0.024), and Cu/Zn ratio (P=007) were observed in late preterm relative to term neonates. Late preterm mothers were significantly younger (P=0.027) than term mothers. Maternal age correlated positively with apo A-I (r=0.424, P=0.012) and negatively with Cu/Zn ratio (r=-0.353, P=0.040). The findings suggest that with daily dietary Centrum Materna supplementation throughout pregnancy, hematological indices were maintained within normal in mothers and neonates, but the levels of microminerals and micromineral ratios were subnormal in late preterm mothers and their neonates.

Associação da concentração plasmática de cobre com metaloproteínas cobre-dependentes em atletas de elite

O cobre é um elemento-traço essencial para a manutenção de vários processos biológicos, tais como metabolismo energético, homeostase de ferro e mecanismos de proteção antioxidante através da atividade da cobre-zinco superóxido dismutase (Cu-Zn SOD), da ceruloplasmina e da metalotioneína. No entanto, o cobre também participa de reações oxidativas que promovem a liberação de radicais livres, podendo prejudicar a integridade e a funcionalidade celular. A atividade física afeta a homeostase do cobre e promove maior utilização de oxigênio, favorecendo a instalação do estresse oxidativo quando mecanismos naturais de proteção antioxidante, incluindo os dependentes de cobre, não atuam adequadamente. Não há relatos na literatura sobre a associação de diferentes níveis de concentração plasmática de cobre com indicadores antioxidantes cobre-dependentes em atletas de elite. O presente estudo objetivou verificar a associação entre diferentes níveis plasmáticos de cobre e metaloproteínas cobre-dependentes, com atividade antioxidante, em atletas de elite. Os indicadores bioquímicos (metalotioneína e Cu-Zn SOD eritrocitárias, ceruloplasmina e cobre plasmáticos) foram avaliados em 50 atletas, homens e adultos, utilizando metodologias já consolidadas. Os resultados mostraram que 32% dos atletas apresentaram níveis de cobre plasmático inferiores a 11µmol/L, 38% entre 11-13µmol/L e 30% &gt; 13µmol/L. As associações encontradas entre cobre plasmático e ceruloplasmina (r = 0,31; p = 0,04) e Cu-Zn SOD (r = 0,32, p = 0,02); metalotioneína eritrocitária e ceruloplasmina (r = 0,40, p = 0,006) e Cu-Zn SOD (0,73, p = 0,001) e entre Cu-Zn SOD e ceruloplasmina (r = 0,37, p &lt; 0,001) demonstraram que a atividade da Cu-Zn SOD e a concentração de metalotioneína eritrocitárias são sensíveis a menor concentração, enquanto que a ceruloplasmina é sensível a elevadas concentrações plasmáticas de cobre, sugerindo que há um equilíbrio homeostático entre antioxidantes, cobre-dependentes, intra e extracelulares em atletas de elite.

Plasma Lipid Peroxidation Zinc and Erythrocyte Cu-Zn Superoxide Dismutase Enzyme Activity in Patients with Type 2 Diabetes Mellitus in Gorgan City (South East of the Caspian Sea)

Plasma Lipid Peroxidation Zinc and Erythrocyte Cu-Zn Superoxide Dismutase Enzyme Activity in Patients with Type 2 Diabetes Mellitus in Gorgan City (South East of the Caspian Sea)

Oxidative stress and antioxidant defense in patients with chronic hepatitis C patients before and after pegylated interferon alfa-2b plus ribavirin therapy

BackgroundOxidative stress could play a role in pathogenesis of hepatitis C virus (HCV) infection. The aim of our study is to determine oxidant/antioxidant status of patients with chronic hepatitis C (CHC), and the effect of pegylated interferon alfa-2b plus ribavirin combination therapy on oxidative stress.MethodsNineteen patients with chronic HCV infection and 28 healthy controls were included in the study. In control and patient groups, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, erythrocyte malondialdehyde (MDA) levels, erythrocyte CuZn-superoxide dismutase (SOD), erythrocyte glutathione peroxidase (GSH-Px) activities were measured. After pegylated interferon alfa-2b and ribavirin combination therapy for 48 weeks, these parameters were measured again in the patient group.ResultsSerum MDA levels increased significantly in CHC patients (n:19), before the treatment when compared with healthy subjects (n:28) 9.28 ± 1.61, 4.20 ± 1.47 nmol/ml, p < 0.001 respectively. MDA concentration decreased significantly (p < 0.001) after the treatment as well as ALT, AST activity, in erythrocytes of these patients. Average antioxidant enzymes (superoxide dismutase and glutathione peroxidase) were significantly lower in erythrocytes of patients with CHC before treatment compared with the control group (both, p < 0.001). Chronic Hepatitis C patients after pegylated interferon alfa-2b and ribavirin therapy showed values of SOD, GSH-Px were significantly higher than pretreatment levels (both, p < 0.001).ConclusionOur results show that patients with chronic HCV infection are under the influence of oxidative stress associated with lower levels of antioxidant enzymes. These impairments return to level of healthy controls after pegylated interferon alfa-2b plus ribavirin combination therapy of CHC patients. Although interferon and ribavirin are not antioxidants, their antiviral capacity might reduce viral load, and inflammation, and perhaps through this mechanism might reduce virus-induced oxidative stress.

Erythrocyte CuZn Superoxide Dismutase Activity Is Decreased in Iron-Deficiency Anemia

Iron and copper are essential microminerals that are intimately related. The present study was performed to determine the effect of iron-deficiency anemia (IDA) and treatment with iron on laboratory indicators of copper status. Hemoglobin, mean corpuscular volume erythrocyte Zn protoporphyrin, serum ferritin, serum copper, serum ceruloplasmin, and erythrocyte CuZn-superoxide dismutase (SOD) activity were studied in 12 adult women with IDA before and after iron treatment for 60-90 d (100 mg/d Fe, as ferric polymaltose) and in 27 women with normal iron status. Prior to treatment with iron, serum copper and ceruloplasmin were not different between the groups and treatment with iron did not affect these measures. IDA women, before and after treatment with iron, presented a 2.9- and 2-fold decrease in erythrocyte CuZn-SOD activity compared to women with normal iron status (p < 0.001). Treatment with iron increased erythrocyte CuZn-SOD activity of the IDA group; however, this change was not statistically significant. In conclusion, CuZn-SOD activity is decreased in IDA. Measurement of this enzyme activity is not useful for evaluating copper nutrition in iron-deficient subjects.

Plasma Lipid Peroxidation Zinc And Erythrocyte Cu-Zn Superoxide Dismutase Enzyme Activity In Patients With Type 2 Diabetes Mellitus In Gorgan City (South East Of The Caspian Sea)

The aim of this study was to determine the changes of plasma lipid peroxidation, zinc and erythrocyte Cu-Zn superoxide dismutase activity in patients with type 2 diabetes mellitus and healthy control in Gorgan city. Fifty type 2 patients with diabetes mellitus and 50 people without diabetes were included in this randomized study. None of patients studied had any diabetic complications. The levels of plasma malondialdehyde and zinc from type 2 diabetes mellitus patients (6.24±0.85 nmol mL-1 and 116.78±5.51 μ dL-1) and control groups (3.63±0.97 nmol mL-1 and 146.86±9.06 μg dL-1) were determined. Erythrocyte Cu-Zn superoxide dismutase activity from patients with diabetes (675.34±60.89 U g-1 Hb) and control groups (1052.70± 52.76 U g-1 Hb) were determined. The increased plasma lipid peroxidation and decreased plasma zinc and erythrocyte Cu-Zn superoxide dismutase activity that we observed in patients with type 2 diabetes mellitus may predispose to the development of cardiovascular complications. We propose that diabetic patients may have elevated requirement for antioxidants. Supplementation with zinc and vitamin or dietary free radical scavengers such as vitamins E and C or tomato, orange and etc. have a potential role in boosting antioxidant-related defences and maybe important in patients with diabetes.

Role of heme oxygenase and bilirubin in oxidative stress in preterm infants.

In a previous study, it was found that the decrease in the total plasma bilirubin level (Btot) in preterm infants was associated with the decrease in oxidative stress. We hypothesized that this occurs as a result of a pro-oxidant effect of heme oxygenase (HO), which outcompetes with the antioxidant properties of bilirubin. In this study we studied 12 preterm infants in whom the plasma levels of Btot, total hydroperoxide (TH), protein SH groups, HO activity, non-transferrin-bound iron (NTBI), and erythrocyte CuZn superoxide dismutase (CuZn SOD) activity were concurrently measured when the Btot was >220 microM and after a Btot drop of >34 microM. The Btot decrease was concurrent with the TH decrease, protein SH groups increase, and the HO and CuZn SOD activity increase and was not associated with an NTBI increase. We concluded that 1) Btot does not exert a meaningful antioxidant effect in vivo; 2) HO does not exert a pro-oxidant effect involving an NTBI increase and that, on the contrary, it could exert an antioxidant effect; and 3) the concurrent HO and CuZn SOD activity increase could indicate a synergic antioxidant effect of the two enzymes.

Superoxide dismutase targets NO from GSNO to Cysbeta93 of oxyhemoglobin in concentrated but not dilute solutions of the protein.

The role of hemoglobin (Hb) in transmitting the vasodilatory property of NO throughout the vascular system is of much current interest. NO exchange between Hb and low-molecular-weight nitrosothiols such as S-nitrosoglutathione (GSNO) has been speculated and reported in vitro. Previously, we reported that NO delivery from GSNO to Cysbeta93 of human oxyHb is prevented in the presence of the Cu chelators, neocuproine, and DTPA.(1) In the present work, 5 mM solutions of commercial human Hb were found by ICP-MS to contain approximately 20 microM Cu and Zn, suggesting the presence of Cu,Zn-superoxide dismutase (CuZnSOD), which was confirmed by Western blotting. SOD activity measurements were consistent with the presence of approximately 20 microM CuZnSOD monomer in 5 mM Hb solutions, which is the physiological concentrations of these proteins in the red blood cell. Incubation of 3.75 mM oxyHb (15 mM heme; 7.5 mM Cysbeta93) with 3.75 or 7.5 mM GSNO gave rise to 50% or 100% S-nitrosation, respectively, of Cysbeta93 as monitored by FTIR nu(SH) absorption, whereas excess GSNO over Cysbeta93 converted oxyHb to metHb due to the reaction, oxyHb + NO<==>metHb + NO(3)(-). Removal of CuZnSOD by anion-exchange chromatography yielded an oxyHb sample that was unreactive toward GSNO, and replacement with bovine CuZnSOD restored reactivity. Addition of 1 microM GSNO (Cysbeta93/GSNO = 1) to solutions diluted 10(4)-fold from physiological concentrations of oxyHb and CuZnSOD resulted largely in metHb formation. Thus, this work reports the following key findings: CuZnSOD is an efficient catalyst of NO transfer between GSNO and Cysbeta93 of oxyHb; metHb is not detected in oxyHb/GSNO incubates containing close to the physiological concentration (5 mM) of Hb and CuZnSOD when the Cysbeta93/GSNO molar ratio is 0.5 to 1.0, but metHb is detected when the total Hb concentration is low micromolar. These results suggest that erythrocyte CuZnSOD may play a critical role in preserving the biological activity of NO by targeting it from GSNO to Cysbeta93 of oxyHb rather than to its oxyheme.

Nitric oxide synthase inhibition by L-NAME in streptozotocin induced diabetic rats: impacts on oxidative stress.

The effects of nitric oxide synthase (NOS) inhibition by Nw-nitro-L-arginine methyl ester (L-NAME) administration on oxidative stress parameters were investigated in streptozotocin (STZ) induced diabetic rats. Lipid peroxidation as reflected by thiobarbituric acid reactive substances (TBARS) was insignificantly higher in diabetic rats. Plasma NO2+NO3 values (p < 0.05) and erythrocyte CuZn superoxide dismutase (CuZn SOD) and glutathione peroxidase (GSH Px) activities were significantly higher (p < 0.01, p < 0.001, respectively) in diabetic rats. L-NAME administration to diabetic rats caused significantly lower CuZn SOD and GSH Px activities (p < 0.01) and NO2+NO3 values (p < 0.001), whereas a significantly higher GSH level (p < 0.01). TBARS/GSH ratio was significantly higher in diabetic rats than controls (p < 0.05) and significantly lower in L-NAME administered diabetic rats than diabetic rats (p < 0.05). This experimental study highlightens the importance of NOS inhibition by L-NAME in the attenuation of oxidative stress in STZ diabetic rats.

Oxidant/antioxidant status in blood of patients with malignant breast tumour and benign breast disease.

The aim of this study was to investigate the alterations in lipid peroxidation and antioxidant enzyme defences in the blood of patients with malignant breast tumour and benign breast disease. Forty patients with malignant breast tumour, 20 patients with benign breast disease and also 20 healthy control subjects were recruited for the study. Malondialdehyde levels in plasma and erythrocytes, and the activities of erythrocyte CuZn-superoxide dismutase, catalase, glutathione peroxidase and glucose-6-phosphate dehydrogenase were measured. Malondialdehyde levels were higher in patients with both benign breast disease and malignant breast tumour compared with control subjects. The activities of all antioxidant enzymes were higher in patients with malignant breast tumour, while only glutathione peroxidase and CuZn-superoxide dismutase activities were higher in patients with benign breast disease. Except for glucose-6-phosphate dehydrogenase, the antioxidant enzymes studied correlated positively with the malondialdehyde levels in patients with malignant breast tumour. On the other hand, only glucose-6-phosphate dehydrogenase activity was increased by the level of malignancy. The activity increases in erythrocyte antioxidant enzymes may be a compensatory upregulation in response to increased oxidative stress especially in patients with malignant breast tumour.

Plasma and erythrocyte levels of trace elements and related antioxidant enzyme activities in low-birthweight infants during the early postnatal period.

To discover the relationship between antioxidant enzyme activities and trace elements in low-birthweight infants during the early postnatal period, we analysed catalase (CAT), CuZn-superoxide dismutase (CuZn-SOD) and glutathione peroxidase (GSH-Px) activities in erythrocytes. and compared them with Fe, Cu. Zn and Se levels in plasma, and Cu, Zn and Se levels in erythrocytes until 16 wk after birth. Thirteen low-birthweight infants whose mean birthweight and gestation were 1520 +/- 293 g and 32.0 +/- 2.8 wk were enrolled in this study. All infants were without chronic complications, well nourished, and predominantly fed standard formula or preterm formula based on cow's milk, commercially available in Japan. Cu and Zn levels in erythrocytes did not decline after birth, in contrast to a temporal decrease in plasma Zn. Erythrocyte CAT activity was significantly higher at 16 wk than that at birth or 4 wk of age. Erythrocyte CuZn-SOD activity did not change throughout the study period. Only Se in plasma and erythrocytes decreased remarkably after birth, which resulted in a significant decline in erythrocyte GSH-Px activity at 8 and 16 wk (11.2 +/- 2.0 and 11.0 +/- 1.1 U/g Hb) compared to that at birth (12.4 +/- 2.1 U/g Hb). During the early postnatal period. erythrocyte CAT and CuZn-SOD activities did not decrease in formula-fed, low-birthweight infants. Japanese commercial formula not supplemented with Se, however, caused postnatal plasma and erythrocyte Se deficiency and a concomitant decline in erythrocyte GSH-Px activity in the same subjects.

Plasma and erythrocyte levels of trace elements and related antioxidant enzyme activities in low‐birthweight infants during the early postnatal period

To discover the relationship between antioxidant enzyme activities and trace elements in low‐birthweight infants during the early postnatal period, we analysed catalase (CAT), CuZn‐superoxide dismutase (CuZn‐SOD) and glutathione peroxidase (GSH‐Px) activities in erythrocytes, and compared them with Fe, Cu, Zn and Se levels in plasma, and Cu, Zn and Se levels in erythrocytes until 16 wk after birth. Thirteen low‐birthweight infants whose mean birthweight and gestation were 1520 ± 293 g and 32.0 ± 2.8 wk were enrolled in this study. All infants were without chronic complications, well nourished, and predominantly fed standard formula or preterm formula based on cow's milk, commercially available in Japan. Cu and Zn levels in erythrocytes did not decline after birth, in contrast to a temporal decrease in plasma Zn. Erythrocyte CAT activity was significantly higher at 16 wk than that at birth or 4 wk of age. Erythrocyte CuZn‐SOD activity did not change throughout the study period. Only Se in plasma and erythrocytes decreased remarkably after birth, which resulted in a significant decline in erythrocyte GSH‐Px activity at 8 and 16 wk (11.2 ± 2.0 and 11.0 ± 1.1 U/g Hb) compared to that at birth (12.4 ± 2.1 U/g Hb). Conclusion: During the early postnatal period, erythrocyte CAT and CuZn‐SOD activities did not decrease in formula‐fed, low‐birthweight infants. Japanese commercial formula not supplemented with Se, however, caused postnatal plasma and erythrocyte Se deficiency and a concomitant decline in erythrocyte GSH‐Px activity in the same subjects.

Superoxide dismutase and glutathione peroxidase function in progressive myoclonus epilepsies

Progressive myoclonic epilepsies (EPM) are difficult to treat and refractory to most antiepileptic drugs. Besides epilepsy, EPMs also involve continuous neurological deterioration. Oxidative stress is thought to be an important factor in this process. We therefore analyzed a series of antioxidant enzymes in the blood of patients and compared with healthy age matched controls. In addition patients were given high doses of N-acetylcysteine (NAC), a glutathione percursor to determine if symptoms of EPM would improve. Five patients, four with EPM 1 (Unverricht–Lundborg disease) and one patient with EPM2 (Lafora body disease) were treated with 6 g/day of NAC. Before treatment, plasma samples were analyzed for glutathione peroxidase activity, catalase activity, extracellular superoxide dismutase (SOD) and CuZn–SOD and compared with the controls. Erythrocyte CuZn–SOD was significantly lower in the EPM patients compared to controls. NAC improved markedly and stabilized the neurological symptoms in patients with EPM 1 but had a doubtful effect in the patient with EPM 2.

Copper-zinc superoxide dismutase from the marine yeast Debaryomyces hansenii.

We have isolated the cytosolic form of Cu-Zn superoxide dismutase (SOD) from the marine yeast Debaryomyces hansenii. This enzyme has a subunit mass of 18 kDa. The preparation was found to be heterogeneous by IF electrophoresis with two pI ranges: 5.14-4.0 and 1.6-1.8. The enzyme preparation had a remarkably strong stability at pH 6.0-7.0, surviving boiling for 10 min without losing more than 60% of activity. On Western blots, this enzyme was recognized by antibodies raised in rabbits against D. hansenii extracts, while only a weak cross-reaction could be detected using antibodies generated against either Saccharomyces cerevisiae or bovine erythrocyte Cu-Zn SODs. In sequencing analysis, a peptide obtained by trypsin digestion was found to have 85% identity to the S. cerevisiae Cu-Zn SOD.

Blood antioxidant defense system and dietary survey of elderly diabetic men

Levels of antioxidants, activities of free radical scavenging enzymes and extent of lipid peroxidation were determined in the blood of 37 elderly diabetic men and 30 control elderly men, 16 without cardiovascular disease (CVD) and 14 with CVD. The mean ±S.D. of the ages of the diabetic men was 66±5 and those of the control men was 69±5, while serum glucose levels of diabetic men were 213±81 mg/dl and that of control subjects were 95±14 mg/dl. Among the diabetic men, 13 men were obese with body mass index>30, 26 men had poor control of diabetes (glycohemoglobin>7%) and 25 men had retinopathy. The diets of the control and diabetic men were evaluated. Blood samples were collected and analyzed for major endogenous antioxidant defense parameters and lipid peroxidation. The results show that diabetic men had significantly lower blood reduced glutathione levels ( p<0.001) and erythrocyte (RBC) CuZn-superoxide dismutase activity ( p<0.001) when compared to control groups with or without CVD. There was no significant differences in plasma vitamin E levels and the activities of catalase and glutathione peroxidase in RBC among the three groups. The extent of lipid peroxidation was highest in diabetic patients, intermediate in controls with CVD, and lowest in controls without CVD. The results suggest that a decline of endogenous antioxidant defense capability contributes to oxidative stress in the diabetic elderly patients. Dietary survey showed that there were no differences in the nutrient intakes of diabetic and control groups. It appears that individual dietary advice is needed for a large portion of diabetic patients in view of their poor glycemic control, hypertriglyceridemia and obesity.

CuZn-superoxide dismutase, extracellular superoxide dismutase, and glutathione peroxidase in blood from individuals homozygous for Asp90Ala CuZu-superoxide dismutase mutation.

The Asp90Ala CuZn-superoxide dismutase mutation is associated with amyotrophic lateral sclerosis (ALS) in both homo- and heterozygous form. We analyzed antioxidant enzymes in blood from 44 individuals homozygous and 114 individuals heterozygous for the Asp90Ala mutation as well as 66 blood relatives carrying the wild-type allele only. Erythrocyte CuZn-superoxide dismutase activity was reduced by 9% in the homozygous individuals, confirming previous findings on a smaller cohort. The specific activity of Asp90Ala mutant CuZn-superoxide dismutase in erythrocytes was equal to that of isolated mutant enzyme and slightly higher than that of isolated wild-type enzyme. There was no evidence for the presence of inactive mutant molecules in erythrocytes, and the lower activity is due to the occurrence of fewer active molecules. There were no significant differences between the groups in plasma extracellular superoxide dismutase content, and the erythrocyte glutathione peroxidase activities were virtually identical. Also, there were no differences in these parameters between homozygous individuals with or without ALS. There was no evidence for any association with ALS of a polymorphic extracellular superoxide dismutase mutation, Arg213Gly. The absence of response of the blood antioxidant enzymes to the Asp90Ala CuZn-superoxide dismutase mutation does not support the theory that the ALS-linked CuZn-superoxide dismutase mutations cause disease by increased oxidant stress.

Antioxidant Status of Breast Cancer and Benign Breast Diseases

This study, designed to investigate antioxidant the erythrocyte anti-oxidant status in malignant and benign breast tumours, was carried out on 38 patients. Based upon their histopathology, the patients were grouped as breast carcinoma (n=23) and benign breast disease (n=15). Of the 23 patients with breast carcinoma 14 had only ductal, 2 had lobular and 7 had ductal + lobular carcinoma. 14 patients with breast carcinoma had axillary metastasis and 9 had no metastasis. Patients in the malignant, group were aged between 38 and 80 years (58.61±13.38); those in the benign group were aged between 20 and 46 years (37±7.42). Heparinised blood samples, drawn preoperatively after an overnight fast, were analysed for erythrocyte CuZn superoxide dismutase (CuZn, SOD), glutathione peroxidase (GSH Px) and glutathione (GSH) values. Erythrocyte GSH-Px values were found to be significantly higher in the malignant group in comparison with the benign group (0.01 >P>0.001 ). CuZn SOD activity and GSH concentration were found to be higher , but not at significant levels, in the malignant group. Measuring the tissue levels of antioxidant status components would serve better for the evaluation of antioxidant profiles in patients with breast carcinoma and benign breast diseases.