Oxygen Free Radical Activity Research Articles

Lipid Peroxidation and Protein Turnover after Trauma and Cold Treatment in Skeletal Muscle of Exercise-trained Rats.

Objective: To examine the effects of muscle trauma and cold application on indicators of oxygen free radical (OFR) and protein levels in exercise trained rat skeletal muscle. Design: Randomized intervention study following an exercise training program. Skeletal muscle tissue analysis 72 hours after trauma and cold treatments. Setting: University animal research laboratory. Subjects: 70 female Sprague-Dawley rats weighing 200-250 grams, housed in wire cages, provided with rat food and water ad libitum. Interventions: 35 rats took part in a daily running program for 8 weeks. After the training program, 28 rats were randomly assigned to one of four groups to study indices of OFR levels after trauma and cold treatments. Further, 28 rats were randomly assigned to one of four groups to study muscle protein content after trauma and cold treatment. A sedentary group (N=7) was included in each analysis as a control group. Groups that received trauma were subjected to a single impact blunt trauma to the medial aspect of the gastrocnemius muscle of the left leg. Following the trauma procedure, groups that received cold treatment had 6 cold applications for 10 minutes, twice per day over a 72 hour period with a 3 × 2 inch gel cold pack, stored at -4°C. Main outcome measures: Medial gastrocnemius muscles were analyzed at 72 hours post-trauma for citrate synthase, indicators of OFR (malonaldehyde and lipofuscin) and protein content. Results: After the endurance training program, significant increases (p<0.05) in citrate synthase activity and protein levels were elicited, but the indices of lipid peroxidation (OFR indicators) were unaltered when compared to sedentary and exercise groups. Following trauma, the net loss of pre-existing protein was statistically significant (p<0.05) and lipid peroxidation activity increased slightly, but not significantly. Further, modulations in lipid peroxidation and protein content were induced in muscle by cold treatment followed by trauma (p<0.05) as the application of cold prevented or delayed indicators of OFR activity and protein loss in the medial gastrocnemius muscle. Conclusions: Cold treatment applied to traumatized muscle significantly decreased lipofuscin content (indices of OFR) and offset the catabolic response in protein content during the inflammatory phase. The early application of cold immediately after trauma is recommended within the protocol guidelines.

Association of Renal Injury with Increased Oxygen Free Radical Activity and Altered Nitric Oxide Metabolism in Chronic Experimental Hemosiderosis

Chronic iron (Fe) overload is associated with a marked increase in renal tissue iron content and injury. It is estimated that 10% of the American population carry the gene for hemochromatosis and 1% actually suffer from iron overload. The mechanism of iron overload-associated renal damage has not been fully elucidated. Iron can accelerate lipid peroxidation leading to organelle membrane dysfunction and subsequent cell injury/death. Iron-catalyzed generation of reactive oxygen species (ROS) is responsible for initiating the peroxidatic reaction. We investigated the possible association of oxidative stress and its impact on nitric oxide (NO) metabolism in iron-overload-associated renal injury. Rats were randomized into Fe-loaded (given 0.5 g elemental iron/kg body weight as iron dextran; IV), Fe-depleted (given an iron-free diet for 20 weeks), and control groups. Renal histology, tissue expression of endothelial and inducible nitric oxide synthases (eNOS and iNOS), renal tissue expression of nitrotyrosine, plasma, and renal tissue lipid peroxidation product, malondialdehyde (MDA), and plasma and urinary NO metabolites (NOx) were examined. Iron overload was associated with mild proteinuria, tissue iron deposition together with significant glomerulosclerosis, tubular atrophy, and interstitial fibrosis. Rare focal glomerulosclerosis and tubulointerstitial changes were noted in normal controls. No renal lesions were observed in Fe-depleted rats. Iron deposits were seen in glomeruli, proximal tubules, and interstitium. The iron staining in the distal tubules was negligible. Both plasma and renal tissue MDA and renal tissue nitrotyrosine were increased significantly in Fe-loaded rats compared with control rats. In contrast, Fe-depleted animals showed a marked reduction in plasma and renal tissue MDA and nitrotyrosine together with significant elevation of urinary NOx excretion. In addition, iron-overload was associated with up-regulation of renal eNOS and iNOS expressions when compared with the control and Fe-depleted rats that showed comparable values. In conclusion, chronic iron overload resulted in iron deposition in the glomeruli and proximal tubules with various renal lesions and evidence of increased ROS activity, enhanced ROS-mediated inactivation, and sequestration of NO and compensatory up-regulation of renal eNOS and iNOS expressions. However, iron depletion was associated with reduced MDA and tissue nitrotyrosine abundance, increased urinary NOx excretion, normal nitric oxide synthase (NOS) expression, and absence of renal injury. These findings point to the possible role of ROS in chronic iron overload-induced renal injury.

Lipid peroxidation in umbilical arterial blood at birth: the effects of breech delivery.

To determine oxygen free radical activity in breech presentation at birth and relate it to umbilical cord blood acid-base status. A series of 63 singleton deliveries (28 cephalic deliveries as controls), 23 breech deliveries with normoacidemia, and 12 breech deliveries with mild acidaemia) had determination of malondialdehyde and acid-base parameters. The delivery suite in the Department of Obstetrics and Gynaecology at the Süleyman Demirel University, Isparta, Turkey. Sixty-three singleton infants born at term with spontaneous or induced labour and initially normal fetal heart rate tracing. After delivery, umbilical cord arterial and venous blood samples were collected for the determination of malondialdehyde concentrations. Oxygen saturation, pO2, pCO2, pH, and base excess were also measured. Umbilical cord arterial and venous blood gases and malondialdehyde levels. There was a significant correlation between umbilical arterial and venous levels of malondialdehyde and all acid-base parameters (P < 0.001). There were negative correlations between malondialdehyde levels and pH, pO2 and bicarbonate, while there was a positive correlation between malondialdehyde concentrations and pCO2. A positive correlation between malondialdehyde levels and base excess was present in the control group and total breech group (n = 35). The malondialdehyde levels in the total breech group, nonacidaemic breech group (n = 23) and the mildly acidaemic breech group (n = 12) were significantly higher than those in the control group (P < 0.0001). However, acid-base parameters in the nonacidaemic breech group were not statistically different from those in the control group. Lipid peroxidation products (malondialdehyde) existed to some extent in the umbilical cords of newborns with normal acid-base parameters in breech delivery. Our data support the contention that lipid peroxide may be a more sensitive measure for metabolic dysfunctions due to fetal hypoxia than acid-base balance.

Indicators of oxidative injury and alterations of the cell membrane in the skeletal muscle of rats submitted to ischemia and reperfusion.

Oxygen free radicals are considered to be important components involved in the physiopathological tissue alterations observed during ischemia and reperfusion. The objective of the present study was to investigate oxidative stress based on indicators of oxygen free radical activity and on the changes in behavior of the lipoprotein membrane (O-phosphoserine) in the skeletal muscle of rats. Twenty Wistar rats were divided into two groups of 10. One group was submitted to 3 h of total ischemia by applying a tourniquet to the hind limb and the contralateral hind limb was used as control. The second group was submitted to the same procedure and was reperfused for 45 min after 3 h of ischemia by removing the tourniquet, where the contralateral hind limb of the same animal was used as control. Muscle biopsies were taken after ischemia and reperfusion and the parameters indicating oxidative stress (reduced and oxidized glutathione, malondialdehyde, glutamine synthetase, protein carbonyl) and O-phosphoserine (OPS) alterations were analyzed. The following results display control versus experimental hindlimbs groups obtained from the same animal. The skeletal muscle of rats submitted to total ischemia of 3 h duration showed increased OPS release (2.69 +/- 4.52 vs 8.03 +/- 7.20; n = 10; P = 0.024) and no change in reduced and oxidized glutathione, glutamine synthetase, protein carbonyl, or malondialdehyde. After 45 min of reperfusion there was an increase in oxidized glutathione levels (0.30 +/- 0.06 vs 0.39 +/- 0.09; n = 8; P = 0.02) and malondialdehyde levels (154. 78 +/- 26.13 vs 206.30 +/- 47.30; n = 9; P = 0.008), a fall in glutamine synthetase (21.80 +/- 3.61 vs 13.52 +/- 6.78; n = 9; P = 0. 004), and a return of OPS to levels close to the initial ones. No changes in reduced glutathione or protein carbonyl were observed in the two groups studied. After a total ischemia duration of 3 h there were signs of damage to the phospholipid membrane of the rat skeletal muscle, as demonstrated by the elevation of OPS and the few or no oxidative changes in the cell. After 45 min of reperfusion, oxidative damage to the lipoprotein components of the cell membrane was observed, characterized by elevations of oxidized glutathione and malondialdehyde levels and a fall in glutamine synthetase levels.

Role of Src in the Modulation of Multiple Adaptor Proteins in FcαRI Oxidant Signaling

AbstractCross-linking of Fc receptors for IgA, FcαR (CD89), on monocytes/macrophages is known to enhance phagocytic activity and generation of oxygen free radicals. We provide evidence here that the FcαR signals through the γ subunit of FcɛRI in U937 cells differentiated with interferon γ (IFNγ). Our results provide the first evidence that FcαR-mediated signals modulate a multimolecular adaptor protein complex containing Grb2, Shc, SHIP, CrkL, Cbl, and SLP-76. Cross-linking of FcαRI using anti-FcαRI induces the phosphorylation of the γ subunit as detected by mobility retardation on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Stimulation of FcαRI induced the tyrosine phosphorylation of Shc and increased the association of Grb2 with Shc and CrkL. Grb2 associates constitutively with Sos, and the latter undergoes mobility shift upon FcαRI stimulation. The complex adapter proteins, Cbl and SLP-76, are physically associated in myeloid cells and both proteins undergo tyrosine phosphorylation upon FcαR stimulation. These data indicate that the stimulation of FcαR results in the modulation of adaptor complexes containing tyrosine-phosphorylated Cbl, Shc, SHIP, Grb2, and Crkl. Experiments performed with the Src kinase inhibitor, PP1, provide the first evidence that Src kinase activation is required for FcαRI-induced production of superoxide anions and provide insight into the mechanism for FcαR-mediated activation of downstream oxidant signaling in myeloid cells.

Role of Src in the Modulation of Multiple Adaptor Proteins in FcRI Oxidant Signaling

Cross-linking of Fc receptors for IgA, FcR (CD89), on monocytes/macrophages is known to enhance phagocytic activity and generation of oxygen free radicals. We provide evidence here that the FcR signals through the γ subunit of FcɛRI in U937 cells differentiated with interferon γ (IFNγ). Our results provide the first evidence that FcR-mediated signals modulate a multimolecular adaptor protein complex containing Grb2, Shc, SHIP, CrkL, Cbl, and SLP-76. Cross-linking of FcRI using anti-FcRI induces the phosphorylation of the γ subunit as detected by mobility retardation on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Stimulation of FcRI induced the tyrosine phosphorylation of Shc and increased the association of Grb2 with Shc and CrkL. Grb2 associates constitutively with Sos, and the latter undergoes mobility shift upon FcRI stimulation. The complex adapter proteins, Cbl and SLP-76, are physically associated in myeloid cells and both proteins undergo tyrosine phosphorylation upon FcR stimulation. These data indicate that the stimulation of FcR results in the modulation of adaptor complexes containing tyrosine-phosphorylated Cbl, Shc, SHIP, Grb2, and Crkl. Experiments performed with the Src kinase inhibitor, PP1, provide the first evidence that Src kinase activation is required for FcRI-induced production of superoxide anions and provide insight into the mechanism for FcR-mediated activation of downstream oxidant signaling in myeloid cells.

Lipid peroxidation in cord blood at birth: the effect of labour.

To determine the effect of labour on free oxygen radical activity in the fetus, as reflected by lipid peroxide levels in umbilical cord arterial blood. Prospective, observational study. Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong. Umbilical cord arterial and venous blood samples were collected from singleton term infants delivered by elective caesarean section. Base excess, PO2, pCO2 and pH were measured in both samples and compared to identify double venous samples. Cord arterial acid-base balance and concentrations of organic hydroperoxides and malondialdehyde were compared with those obtained from normal vaginal deliveries. Cord arterial blood samples, obtained from cases of uncomplicated labour followed by spontaneous vaginal delivery, had significantly higher lipid peroxide concentrations than those delivered following elective caesarean section. This was most marked for malondialdehyde with a median value increased by 105%, whilst organic hydroperoxide was increased by only 27%. Of the acid-base parameters, base excess was increased by 78%, with only minimal changes in pH, pCO2 and PO2. These differences remained highly significant after including other pregnancy characteristics in multivariate analysis. The findings indicate that high levels of free oxygen radical activity in the fetus are a function of the labour process, as are changes in acid-base balance.

Role of increased oxygen free radical activity in the pathogenesis of uremic hypertension

Earlier studies have demonstrated increased oxygen free radical (OFR) activity, diminished antioxidant capacity and reduced OFR-inactivating enzymes in chronic renal failure (CRF). Via inactivation of nitric oxide (NO), oxidation of arachidonic acid and a direct vasoconstrictive action, OFR can potentially raise blood pressure (BP). This study was designed to test the hypothesis that increased OFR activity may contribute to CRF hypertension. Four weeks after 5/6 nephrectomy rats were treated for two weeks with either lazaroid, a potent antioxidant and lipid peroxidation inhibitor (CRF-LZ group), or vehicle alone (CRF group) by daily gastric gavage. The control group was sham operated and placebo treated. The CRF group exhibited significant increases in BP and plasma lipid peroxidation product, malondialdehyde (MDA), indicating enhanced OFR activity. This was accompanied by decreased urinary nitrate/nitrite (NOx) excretion suggesting depressed NO production. LZ therapy normalized plasma MDA and significantly ameliorated CRF-induced hypertension. Both MDA and blood pressure (BP) rose to values seen in the untreated CRF group within two weeks after termination of LZ therapy. Intravenous administration of the hydroxyl radical scavenger, dimethylthiourea (DMTU), significantly lowered BP and raised urinary NOx excretion. However, no discernible effects were found with either superoxide dismutase or catalase (superoxide and H2O2 quenchers). The results suggest that increased OFR activity is, in part, responsible for CRF-associated HTN. The study further points to hydroxyl radicals as the major source of OFR in CRF animals. If substantiated in humans, antioxidant therapy becomes a logical adjunct in the management of CRF.

Monitoring of oxygen free radical activity in the acute phase after severe head injury

Neurosurgical ICU, Hospital of Treviso; Department of Neurosurgery, Humanitas Clinic, Rozzano (MI); and Institute of Pharmacology, University of Pavia, Italy

Monitoring of oxygen free radical activity in the acute phase after severe head injury

Paolin, A.; Nardin, L.; Gaetani, P.; Rodriguez, R.; Finotti, N.; Marzatico, F. Author Information

Impact of oxygen therapy on antioxidant status in newborns. Relationship with infection risk.

Newborns requiring intensive clinical care are susceptible to a wide range of excessive oxygen free radical production-related problems. In utero, fetal organs, particularly lungs, are exposed to relatively hypoxic tensions which rise abruptly after birth and this transition may cause oxidative injury in the neonate. The aims of this study were to determine oxygen free radical activity in neonates at the first 24 h, examine the role of immaturity and infection risk and compare the degree of oxidant stress in newborns treated with different oxygen concentrations. Plasma selenium levels in neonates with high infection risk (IR) were significantly lower than in healthy neonates. Comparative study of selenium in preterm, term and young infants showed age-related increases and differences were significant. Plasma selenium values were lower when oxygen therapy was administered. Vitamin E levels were significantly decreased in IR compared with healthy newborns. The results suggest that selenium and vitamin E deficiencies predispose to neonatal infection and that supplementary oxygen contributes significantly to decreasing the antioxidant defence system.

Altered nitric oxide metabolism and increased oxygen free radical activity in lead-induced hypertension: Effect of lazaroid therapy

Chronic exposure to low levels of lead results in sustained hypertension (HTN) in humans and experimental animals. The mechanism of lead-induced HTN remains unclear. We investigated the possible role of reactive oxygen species (ROS) and their impact on nitric oxide (NO) metabolism in lead-induced HTN. Male Sprague-Dawley rats were treated with lead (100 ppm in drinking water) for twelve weeks. They were then treated with either the potent antioxidant, lazaroid (des-methyl-tirilazad, 5 mg/kg i.p., twice daily) (Pb-Lz group) or placebo (Pb group) for two weeks and monitored for an additional two weeks. A group of normal animals served as controls (N = 6 in each group). Lead administration resulted in marked HTN together with a significant rise in plasma concentration of lipid peroxidation product, malondialdehyde (MDA, reflecting increased ROS generation) and a twofold reduction in urinary excretion of NO metabolites, that is, total nitrates and nitrites (NOx). Lazaroid therapy led to prompt normalization of blood pressure, plasma MDA and urinary NOx. In contrast, blood pressure and plasma MDA remained elevated, and recovery of urinary NOx excretion was slow with placebo therapy. No significant difference was found in creatinine clearance between the study groups during the observation period. Thus, chronic lead exposure resulted in marked HTN coupled with increased ROS production and decreased urinary NOx excretion. Administration of the potent antioxidant, lazaroid, abrogated HTN and reversed the abnormalities of plasma MDA and urinary NOx excretion, thus supporting the role of ROS in lead-induced HTN in this model.

Oxygen free radical activity in the second stage of labor.

To assess fetal cellular injury arising from oxygen free radical activity in relation to the duration of the second stage of labor. Cord arterial pH, malondialdehyde and organic hydroperoxides levels were determined following vaginal delivery of 326 term singleton pregnancies. Of these, 35 (11%) received epidural analgesia. The length of the second stage was recorded from the time of full dilatation to delivery. Arterial cord pH, malondialdehyde and organic hydroperoxides were significantly correlated with duration of second stage of labor (r=-0.4492; r=0.2542; r=0.2244; respectively, p<0.001). The association of lipid peroxidation products with second stage duration was independent of cord pH. This correlation was unchanged when cases of operative delivery were excluded. However, the relationship lost statistical significance amongst cases who received epidurals. The duration of the second stage is correlated with raised reactive oxygen species-derived lipid peroxidation products. Caution should be exercised in the management of prolonged second stage of labor.

Lipid peroxidation in cord blood: the effects of umbilical nuchal cord.

To examine the relation between cord entanglement around the fetal neck and umbilical arterial measures of free oxygen radical activity at birth. Prospective observational study of the lipid peroxide levels in umbilical cord arterial blood. Delivery suite of Prince of Wales Hospital, Hong Kong. Women with a singleton pregnancy, at term (37 to 42 weeks of gestation), cephalic presentation, spontaneous or induced labour and an initially normal fetal heart rate tracing. A series of 230 singleton deliveries had determination of two lipoperoxides in umbilical arterial cord blood: malondialdehyde and organic hydroperoxide. Umbilical pH, PO2, PCO2 and base excess were also measured in both umbilical cord arterial and venous blood. Umbilical cord blood gases and arterial malondialdehyde and organic hydroperoxide levels. Forty-nine cases of nuchal cord were observed: 14 tight and 35 loose. Tight nuchal cord was associated with a low 1-min Apgar score and increased lipid peroxide levels. Loose nuchal cord was associated with larger amniotic fluid volume and significantly decreased lipid peroxide levels. This study suggests that tight nuchal cord is associated with increased free oxygen radical activity in the fetus, but that loose nuchal cord carries no adverse effects.

Lipid peroxidation in cord blood at birth: a marker of fetal hypoxia during labour.

This prospective study examined purine metabolism in relation to free oxygen radical activity, as reflected by lipid peroxide levels in umbilical cord blood at birth. Departments of Obstetrics and Gynaecology and of Chemical Pathology, the Chinese University of Hong Kong, Hong Kong, and Purine Research Laboratory, UMDS of Guy's and St. Thomas' Hospitals, London, UK. Umbilical cord arterial and venous blood samples were collected from 132 singleton term deliveries for determination of hypoxanthine, xanthine, inosine, uric acid, organic hydroperoxides (OHP) and malondialdehyde. Oxygen saturation, PO2, pCO2, pH, and base excess (BE) were also measured. There was a significant correlation between umbilical arterial and venous levels of hypoxanthine, xanthine, inosine, uric acid and all acid-base parameters (p < 0.001). Significant arteriovenous differences were observed for all parameters with the exception of inosine, uric acid and OHP. Umbilical arterial xanthine and potassium correlated significantly with OHP, but hypoxanthine, inosine and uric acid did not. In 13 babies classified as severely asphyxiated at birth (umbilical arterial pH <7.15, BE <-8), xanthine and OHP levels were significantly elevated when compared with non-asphyxiated babies. No significant differences were observed for hypoxanthine, inosine or uric acid. The findings indicate that OHP, either in cord arterial or venous blood, is the best marker of free oxygen radical activity in the fetus, and that this correlates with other evidence of cellular hypoxia-reperfusion injury. We propose OHP is a better measure of perinatal outcome than either acid-base balance or hypoxanthine.

Measurement of oxygen free radicals in a rabbit shock lung model. Effect of superoxide dismutase and retinol.

To develop an isolated rabbit lung model in which oxygen free radical activity could be measured and to examine the effects on the model of oxygen free radical scavengers. Prospective, randomized study. A clinical and basic research facility attached to a teaching hospital. Twenty-five New Zealand white rabbits weighing 3.5 to 4.5 kg. The mechanism of lung injury by oleic acid or by phorbol myristate acetate (20 ng/mL) plus polymorphonuclear neutrophils (PMA-PMN) in an ex vivo rabbit shock lung model may be the production of oxygen free radicals. Using a standard heart-lung preparation from these rabbits, baseline mean pulmonary artery pressure was maintained at 15 mm Hg and the mean airway pressure at 10 mm Hg. Experimental perfusates were infused over 30 minutes, followed by Krebs-Henseleit solution, pH 7.4. Dimethyl pyrroline oxide trapped oxygen free radicals, levels of which were measured by electron paramagnetic resonance spectroscopy. Lung injury was assessed by light and scanning electron microscopy and by lung weight. A 5-fold increase in pulmonary artery pressure (P < .001) and a nearly 3-fold increase in mean airway pressure (P < .001) were observed in both the oleic acid and PMA-PMN models. Superoxide dismutase (20,000 U/kg), but not retinol palmitate (2000 U), prevented lung injury, the increases in pulmonary artery pressure and mean airway pressure, and the increase in oxygen free radicals in the PMA-PMN model. There were no increases in oxygen free radicals in the control, oleic acid, or PMA-PMN/superoxide dismutase groups (n = 5 in each group). Maximum mean +/- SD increases in oxygen free radicals were 112 +/- 22 nmol/L in the PMA-PMN group (P < .003, n = 5) and 108 +/- nmol/L in the PMA-PMN/retinol group (P < .003, n = 5). The mechanism of lung injury in the PMA-PMN model is an increase in oxygen free radicals, because superoxide dismutase prevents both the rise in oxygen free radicals and lung injury. Administration of retinol does not prevent lung injury. Oleic acid produces injury not by an increase in oxygen free radicals but rather by another, unknown mechanism.

Lipid peroxidation in cord blood at birth

OBJECTIVE: The purpose of this study was to determine oxygen free radical activity in the neonate at birth and relate it to umbilical cord blood acid-base status. STUDY DESIGN: A series of 110 singleton deliveries had determination of two lipoperoxides in umbilical cord blood: malondialdehyde and organic hydroperoxide. Umbilical pH, PO 2, PCO 2, bicarbonate, and base excess were also measured. RESULTS: There was a significant association between lipoperoxides and cord blood pH and base excess. A significant difference existed in the levels of umbilical artery lipoperoxides between nonacidemic and acidemic fetus, as defined by an umbilical arterial pH <7.20. CONCLUSION: There is a positive association between lipoperoxide production and acid-base balance at delivery. (A M J O BSTET G YNECOL 1996;174:62-5.)

Neurovascular dysfunction in diabetic rats. Potential contribution of autoxidation and free radicals examined using transition metal chelating agents.

Oxygen free radical activity is elevated in diabetes mellitus and has been implicated in the etiology of vascular complications. Recent studies have shown that impaired perfusion of nerve endoneurium is a major cause of nerve fiber dysfunction in experimental diabetes. Free radical scavenger treatment prevents the development of nerve conduction abnormalities in diabetic rats. In vitro experiments suggest that autoxidation reactions of glucose, catalyzed by free transition metal ions, are a potential source of free radicals in diabetes. We investigated whether chronic treatment with deferoxamine and trientine, transition metal chelating agents which can prevent autoxidation, could correct nerve conduction and blood flow changes in streptozotocin-diabetic rats. A 20% reduction in sciatic nerve motor conduction velocity after 2 mo diabetes was 90% ameliorated by 2 wk of treatment with deferoxamine or trientine. Sciatic endoneurial nutritive blood flow was 45% reduced by diabetes, but was completely corrected by treatment. In contrast, transition metal chelation had no effect on blood flow or conduction velocity in nondiabetic rats. Thus, the data support the hypothesis that increased free radical activity by glucose autoxidation as a result of impaired transition metal handling is a major cause of early neurovascular deficits in diabetes.

Effects of manganese superoxide dismutase on lung fluid balance after smoke inhalation.

There is evidence of increased oxygen free radical activity after smoke inhalation with and without concomitant burn injury. We determined the effects of manganese superoxide dismutase (Mn SOD) on lung fluid balance as measured by lung microvascular permeability coefficient (sigma), filtration coefficient (Kf), and lymph flow. Merino breed ewes (n = 6/group) were surgically prepared. The SOD group (SOD) received Mn SOD (9,000 U/kg) as an intravenous bolus and was insufflated with smoke. The control group (CON) received saline and smoke. sigma and Kf were determined 24 h before and 24 h after smoke injury. Lymph flow, arterial O2-to-inspired O2 fraction ratio, systemic hemodynamics, and pulmonary arterial and capillary pressures were measured. The sigma was significantly (P < 0.05) higher after smoke insufflation in SOD compared with CON (0.71 +/- 0.03 vs. 0.53 +/- 0.05). Kf was significantly lower after smoke insufflation in SOD compared with CON (0.038 +/- 0.010 vs. 0.061 +/- 0.010). Lymph flows were significantly lower during the 24 h after smoke insufflation in SOD compared with CON (33 +/- 7 vs. 55 +/- 8 ml/h at 24 h). Arterial O2-to-inspired O2 fraction ratio was significantly improved at 6 and 12 h after smoke insufflation in SOD compared with CON at the same time points. Mn SOD meliorates the lung microvascular permeability changes associated with smoke inhalation injury.

Evidence of oxidant injury and tubular damage in early diabetic nephropathy

Two groups of patients with insulin-dependent diabetes mellitus of > 10 years duration and either persistent normoalbuminuria (group 1, n = 49; albumin excretion < 30 mg/day) or microalbuminuria (group 2, n = 33; albumin excretion 30-300 mg/day) were investigated for evidence of free oxygen radical activity (erythrocytic superoxide dismutase and glutathione peroxidase) and oxidant injury (serum malondialdehyde). Glomerular proteinuria (albuminuria, transferrinuria), tubular proteinuria (retinol-binding protein) and tubular enzymuria (N-acetyl-glucosaminidase and leucine aminopeptidase) were also measured. Healthy controls (n = 38) were matched for age and sex. Groups 1 and 2 were similar in terms of age, sex, duration of diabetes and recent glycaemic control. Serum cholesterol and creatinine were similar in all three groups. Free-radical activity and oxidant injury were significantly higher in groups 1 and 2 than in controls (p < 0.001). Glomerular proteinuria, tubular proteinuria and enzymuria were significantly higher in group 2 than in group 1 and controls (p < 0.01). Group 1 had significantly higher transferrinuria, tubular enzymuria and tubular proteinuria than controls. However, groups 1 and 2 were similar in degree of free oxygen radical generation and oxidant injury. In diabetic nephropathy, oxidant injury and renal tubular damage accompany and may even precede microalbuminuria. The presence of these abnormalities in the absence of glomerular proteinuria favours the hypothesis that alterations first occur in the peritubular microcirculation, which by causing oxidant injury and tubular damage, may initiate diabetic nephropathy.