B. Matkovics

Involvement of oxygen-derived free radicals in L-arginine-induced acute pancreatitis

This study was aimed at an assessment of the role of oxygen-derived free radicals in the pathogenesis of L-arginine (Arg)-induced acute pancreatitis in rat, by measuring the levels of malonyl dialdehyde (MDA), glutathione peroxidase (GPx), catalase, and superoxide dismutase (Mn- and Cu,Zn-SOD) in the pancreatic tissue, and evaluating the protective effect of the xanthine oxidase inhibitor allopurinol. Acute pancreatitis was induced in male Wistar rats by injecting 2 × 250 mg/100 g body weight of Arg intraperitoneally in a 1-hr interval, as a 20% solution in 0.15 M NaCl. Control rats received the same quantity of glycine. Allopurinol, 100 or 200 mg/kg, was administered subcutaneously 30 min before the first Arg injection. Rats were killed at 6, 12, 24, and 48 hr following Arg administration, and acute pancreatitis was confirmed by a serum amylase level elevation and typical inflammatory features observed microscopically. The serum level of amylase reached the peak level at 24 hr after the Arg injection (30,800 ± 3813 vs 6382 ± 184 units/liter in the control) and normalized at 48 hr. The tissue concentration of MDA was significantly elevated at 24 hr and reached the peak value at 48 hr (5.00 ± 1.75 vs 0.28 ± 0.05 nM/mg protein in the control). The catalase and Mn-SOD activities were significantly decreased throughout the study, while the GPx activity was significantly reduced at 6 and 12 hr, and the Cu,Zn-SOD activity was significantly lower at 12 hr after the Arg injection as compared with the controls. Allopurinol treatment markedly reduced the serum amylase elevation (12.631 ± 2.257 units/liter at 24 hr) and prevented the increase in tissue MDA concentration (0.55 ± 0.09 nM/mg protein at 48 hr). Both doses of allopurinol significantly ameliorated the pancreatic edema, necrosis, and inflammation at 48 hr after Arg administration. Oxygen-derived free radicals are generated at an early stage of Arg-induced acute pancreatitis. Prophylactic allopurinol treatment prevents the generation of reactive oxygen metabolites, reduces the serum amylase concentration, and exerts a beneficial effect on the development of histopathological changes.

Oxidative stress and antioxidant defense mechanism in glomerular diseases

The changes in red blood cell (RBC) lipid peroxidation [measured via the malonyl dialdehyde (MDA) concentration], reduced (GSH), and oxidized glutathione (GSSG) levels, hemoglobin (Hb) oxidation and antioxidant enzyme [catalase (Cat), glutathione peroxidase, and superoxide dismutase (SOD)] activities were studied in 45 pediatric patients with various glomerular diseases [minimal change nephrotic syndrome (MCNS) in relapse or in remission, lupus nephropathy (SLE), poststreptococcal glomerulonephritis (APSGN), IgA nephropathy (IGA gn)], and in 20 adult patients with IGA gn and also in 15 pediatric and 14 adult controls. The in vitro effects of hydrogen peroxide [acetyl phenylhydrazine (APH) test] on the GSH and Hb metabolisms were likewise investigated. There was an increased oxidative stress in MCNS with relapse, IGA gn, SLE gn, and APSGN, which could be detected in the GSH and Hb oxidation and in the lipid peroxidation on the peripheral RBC-s. The RBC SOD and Cat activities were significantly lower in all patients than in the controls. The RBC GSSG level was significantly elevated in all patients, with the exception of MCNS in remission. This stimulated a compensatory GSH production in MCNS with relapse and in IGA gn, but not in SLE or APSGN. The regeneration of GSH from GSSG was reduced in MCNS with relapse, SLE, and IGA gn, but not in APSGN. In remission, the GSH-GSSG redox system normalizes, but in vitro the APH test stimulates an intensive Hb oxidation. In conclusion, there is a correlation between the presence of active glomerular disease and the evidence of oxidative changes in the various parameters measured in peripheral RBCs.

Paraquat and atrazine co-resistance in Conyza canadensis (L.) Cronq.

Abstract A biotype of Conyza canadensis (L.) Cronq. from a vineyard subjected to repeated paraquat and triazine herbicide treatment and a wild type from an adjacent ruderal area were examined for resistance to paraquat and atrazine. CO 2 fixation was reduced by paraquat for 3 hr, but thereafter it was strongly stimulated in the resistant biotype, while that of the susceptible biotype remained inhibited. The fluorescence quenching measurements show that paraquat inhibited both biotypes at the level of isolated chloroplasts. Paraquat quenched fluorescence in intact leaves during the first hour of treatment. Fluorescence quenching ceased in the resistant biotype within 24 hr of paraquat treatment. The I 50 with paraquat in the resistant biotype was 170 times higher than that of the susceptible biotype in measurements of fluorescence induction quenching at 24 hr. There were no chlorophyll bleaching and chlorophyll/protein damage in the resistant biotype incubated with paraquat, in marked contrast to the susceptible biotype. The ultrastructure of the chloroplasts in the paraquat-treated leaves of resistant plants was similar to that of the untreated ones. Plastids of the susceptible biotype were disrupted, with considerable rearrangement of the lamellar system, after paraquat application. Superoxide dismutase, ascorbate peroxidase, catalase, and reduced glutathione levels in the total cell extracts were slightly lower in the resistant biotype, but there was a slightly higher level in the extracts of the susceptible biotype in the presence of paraquat. The paraquat-resistant biotype had co-resistance to atrazine as demonstrated through fluorescence induction measurements. The resistance ratio to atrazine was 300. From these results it was concluded that C. canadensis displays co-resistance to paraquat and atrazine; the data indicate that resistance to herbicides with different modes of action can be developed.

Oxidative stress in distant organs and the effects of allopurinol during experimental acute pancreatitis.

SummaryBackground. The present study was aimed at an assessment of the role of oxygen-derived free radicals in the development of local and systemic manifestations of l-arginine (Arg)-induced acute pancreatitis and at an evaluation of the protective effect of the xanthine oxidase inhibitor allopurinol.Methods. Acute pancreatitis was induced in male Wistar rats by injecting 2×250 mg/100 g body weight of Arg intraperitoneally at an interval of 1 h, as a 20% solution in 0.15 M NaCl. Control rats received the same quantity of glycine. In a third group, 200 mg/kg of allopurinol was administered subcutaneously 30 min before the first Arg injection. Rats were killed at 6, 12, 24, or 48 h following Arg administration. Acute pancreatitis was confirmed by a serum amylase level elevation and typical inflammatory features were observed microscopically. Tissue concentrations of malonyl dialdehyde (MDA), superoxide dismutase (Mn- and Cu,Zn-SOD), glutathione peroxidase (GPx), and catalase were measured in the pancreas, liver, and kidney.Results. The tissue concentration of MDA was significantly elevated in each organ. The activities of Mn-SOD, Cu,Zn-SOD, GPx, and catalase were quickly depleted in the pancreas and kidney, whereas only the Mn-SOD and GPx activities were reduced in the liver after the onset of pancreatitis. Histologic examination revealed acinar cell necrosis in the pancreas, but only mild alterations in the liver and kidney. Allopurinol pretreatment prevented the generation of reactive oxygen metabolites in the pancreas and reduced their formation in the kidney.Conclusion. Oxygen-derived free radicals are generated in the pancreas, liver, and kidney at an early stage of Arg-induced acute pancreatitis. The liver and the kidney, but not the pancreas, are able to defend against oxidative stress. The prophylactic application of allopurinol significantly restrains the generation of free radicals in pancreas and kidney.

A comparative study of superoxide dismutase, catalase and lipid peroxidation in red blood cells from muscular dystrophy patients and normal controls

Abstract The superoxide dismutase and catalase activities and lipid peroxidation of the red blood cells (RBC) were determined in Duchenne muscular dystrophy (DMD) patients and healthy controls, and were found to be significantly higher in the DMD patients.

Comparative antioxidant enzyme study in freshwater fish with different types of feeding behaviour

Comparative studies were made of the activities of Cu,Zn-superoxide dismutase, Mn-superoxide dismutase, catalase and glutathione peroxidase in organ homogenates from two herbivorous (grass carp and silver carp) and three omnivorous fish (barbel, crucian carp and common carp). The protein contents and lipid peroxidation of the organ homogenates were also compared. These comparative measurements provided control values for subsequent toxicological examinations. For the herbivorous fish, the highest total superoxide dismutase activity was observed in the kidney, followed in turn by the liver or spleen. In contrast, the highest total superoxide dismutase activity in the barbel was found in the roe, in the crucian carp in the liver, and in the common carp in the liver too. The quantitative distributions of the two peroxide metabolism enzymes glutathione peroxidase and catalase in the organ homogenates present very varied pictures. The results of the lipid peroxidation are similarly difficult to fit into one framework, even for fish following the same diet. It appears that the enzyme activities mentioned in points 4 and 5, together with the lipid peroxidation, depend not on the species, but on the variety.

Examination of the role of oxygen free radicals in bronchial asthma in childhood

The antioxidant enzyme activities, the lipid peroxidation level, the parameters of glutathione metabolism, and the proportion of haemoglobin oxidation products were determined during the symptom-free period of childhood bronchial asthma. A decreased catalase activity and a significantly reduced glutathione instability were demonstrated as compared to the controls. The results indicate that antioxidant protection of the haemoglobin molecule in asthmatic children is considerably decreased.

Paraquat as an agent affecting antioxidant enzymes of common carp erythrocytes

The effects of paraquat on the superoxide dismutase, catalase, glutathionine peroxidase activities and lipid peroxidation at different times of paraquat exposure of Cyprinus carpio morph L. erythrocytes were studied. Typical characteristics were observed in the changes of the enzyme activities of the erythrocytes after exposure to paraquat. The haemoglobin concentration of common carp haemolysates was decreased by exposure to paraquat.

Oxidative damage of red blood cells in haemolytic uraemic syndrome

Changes in red blood cell (RBC) lipid peroxidation [measured by malonyl dialdehyde (MDA) concentration], glutathione (GSH) metabolism, antioxidant enzyme activities (catalase, superoxide dismutase, glutathione peroxidase) and haemoglobin (Hb) metabolites (metHb, carboxy Hb) were studied in six children with post-enteropathic (D+) haemolytic uraemic syndrome (HUS) and ten controls. The in vitro effect of hydrogen peroxide [acetyl-phenylhydrazine (APH) test] on GSH and Hb metabolism was also investigated. MDA levels were significantly higher and the antioxidant enzyme activities were lower in HUS patients than in the controls (P<0.01). The oxidised glutathione concentration was significantly higher in the patients than in the control children (26.3±12.6 vs. 10.9±1.8 nmol/g Hb. Percentage values of carboxy Hb and metHb were also higher in HUS (P<0.01). Incubation of RBC with APH induced a more pronounced decrease in the concentration of GSH (P<0.001) and a significant increase (P<0.01) in the level of metHb and carboxy Hb in the HUS patients. This suggests that there is reduced RBC GSH stability in HUS. Utilisation of GSH and antioxidant enzymes leads to increased Hb oxidation and haemolysis. The oxidative damage may have an important role in the pathogenesis of haemolytic anaemia in HUS.

Oxidative stress changes in L-arginine-induced pancreatitis in rats

The important role of oxygen radicals in acute experimental pancreatitis was demonstrated by study of the changes in the antioxidant system in the blood, liver, kidney, and pancreas of rats after the administration of a large quantity of L-arginine (L-Arg). The changes in lipid peroxidation and in reduced and oxidized glutathione were followed, as well as the activities of peroxide-decomposing enzymes (glutathione peroxidase and catalase) and H2O2-producing superoxide dismutases. The results demonstrated that “oxidative stress” develops and acute pancreatitis appears rapidly after L-Arg treatment. Oxidative stress symptoms are expressed 24 h after the final treatment. Slow restitution of the studied antioxidant system can be demonstrated as early as after 48 h.