==================================CMR40================================== 40. 1. Free radical pathology on aging and disease. a) laboratory animals, i.e. mice, rats, monkeys, etc. 2. i.e. heavy metals, singlet oxygen, lipids, peroxides, superoxides. 3. Free radical quenchers - natural and artificial. a) Vit C, Vit E, Vit A, selenium, B6, Beta Carotene, zinc b) Superoxide dismutase and glutathione peroxidase. 4. Effects of free radicals on immune system. 1 UI - 87121292 AU - Itoh T ; Kawakami M ; Yamauchi Y ; Shimizu S ; Nakamura M TI - Effect of allopurinol on ischemia and reperfusion-induced cerebral injury in spontaneously hypertensive rats. AB - In spontaneously hypertensive rats, we studied the participation of xanthine oxidase-linked free radical in ischemia and reperfusion-induced cerebral injury, using allopurinol, a xanthine oxidase inhibitor. The loss of righting reflex was noted in some animals after a 4 hour occlusion of bilateral common carotid arteries and 19 of 25 animals died within 72 hours after reperfusion. One hour after reperfusion, the cerebral water content increased significantly, with an increase in sodium content and a decrease in potassium content. In 7 animals treated with oral administrations of allopurinol (200 mg/kg) 24 hours and 1 hour before occlusion, no death was found either during occlusion or after reperfusion, and the loss of righting reflex was noted in only one animal 24-72 hours following reperfusion. The increase in cerebral water content and accompanied changes in electrolyte contents were clearly prevented by allopurinol. These results suggest the possibility that the production of xanthine oxidase-linked free radical participates in cerebral injury due to ischemia and reperfusion in spontaneously hypertensive rats. MH - Allopurinol/*PHARMACODYNAMICS ; Animal ; Brain/DRUG EFFECTS/ METABOLISM ; Cerebral Ischemia/*PREVENTION & CONTROL/ PHYSIOPATHOLOGY ; Cerebrovascular Circulation ; Free Radicals ; Male ; Premedication ; Rats ; Rats, Inbred SHR ; Reflex/DRUG EFFECTS ; Superoxide/METABOLISM ; Xanthine Oxidase/*ANTAGONISTS & INHIBITORS/PHYSIOLOGY SO - Stroke 1986 Nov-Dec;17(6):1284-7 2 UI - 87088814 AU - Murphy ME ; Kehrer JP TI - Free radicals: a potential pathogenic mechanism in inherited muscular dystrophy. AB - Despite years of intensive work, the biochemical defect responsible for the pathogenesis of inherited muscular dystrophy has not been identified either in humans or animal models. This review examines evidence in support of the hypothesis that free radicals may be responsible for muscle degeneration in this disorder. A variety of cellular abnormalities noted in dystrophic muscles can be accounted for by free radical mediated damage. In addition, chemical by-products associated with free radical damage are found in dystrophic muscle tissue from humans and animals with this disease. Various enzymatic antioxidant systems can be enhanced as a normal cellular response to oxidative stress, and such changes are seen both in dystrophic muscle cells and certain other tissues of dystrophic animals. An increased level of free radical damage would follow from either: enhanced production of free radical species, or a deficient component of the cellular antioxidant system, such as vitamin E. The free radical hypothesis of muscular dystrophy can account for data supporting several alternative theories of the pathogenesis of this disease, as well as other observations which have not previously been explained. MH - Calcium/METABOLISM ; Collagen/METABOLISM ; *Free Radicals ; Glycerylphosphorylcholine/METABOLISM ; Membranes/PHYSIOLOGY ; Microcirculation ; Muscles/BLOOD SUPPLY/CYTOLOGY ; Muscular Dystrophy/COMPLICATIONS/*PHYSIOPATHOLOGY ; Review ; Serotonin/ METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Vitamin E Deficiency/COMPLICATIONS SO - Life Sci 1986 Dec 15;39(24):2271-8 3 UI - 87076674 AU - Vissers MC ; Winterbourn CC TI - The effect of oxidants on neutrophil-mediated degradation of glomerular basement membrane collagen. AB - The contribution of activated oxygen species to neutrophil-mediated degradation of basement membrane collagen was investigated. In preliminary experiments, pre-exposure of either albumin or glomerular basement membrane to neutrophil myeloperoxidase with H2O2 and chloride increased their susceptibility to proteolysis 2-3-fold. In the basement membrane model, neutrophils are stimulated by trapped immune complexes to adhere, produce oxidants and degranulate. Degradation, measured as the amount of hydroxyproline solubilised, was due to neutral proteinases, particularly elastase, and depended on cell number and the amount of proteinase released. Experiments with oxidant scavengers and inhibitors and with neutrophils from donors with chronic granulomatous disease or myeloperoxidase deficiency showed that oxidants did not affect degradation of the basement membrane when this was measured on a per cell basis. However, oxidative inactivation of the released granule enzymes occurred. Activities of elastase, beta-glucuronidase and lysozyme were 1.5-2-times higher in the presence of catalase, but were unaffected by superoxide dismutase or hydroxyl radical scavengers. Inactivation did not occur with chronic granulomatous disease or myeloperoxidase deficient neutrophils. When related to the activity of released elastase, or to other degranulation markers, collagen degradation was decreased in the presence of catalase, or with chronic granulomatous disease or myeloperoxidase deficient cells. This implies that the basement membrane was made more digestible by myeloperoxidase-derived oxidants, as occurred in the cell-free experiments. Taken together, the results indicate that neutrophil oxidants have two opposing effects. They increase the susceptibility of the collagen to proteolysis and inactivate the proteinases responsible. MH - Basement Membrane/*METABOLISM ; Chlorides/*PHARMACODYNAMICS ; Collagen/*METABOLISM ; Cytoplasmic Granules/ENZYMOLOGY/SECRETION ; Free Radicals ; Granulomatous Disease, Chronic/PATHOLOGY ; Human ; Hydrogen Peroxide/*PHARMACODYNAMICS ; IgG ; Kidney Glomerulus/*DRUG EFFECTS ; Myeloperoxidase/DEFICIENCY/METABOLISM ; Neutrophils/*DRUG EFFECTS/ENZYMOLOGY/SECRETION ; Oxidation-Reduction ; Peptide Hydrolases/METABOLISM/SECRETION ; Support, Non-U.S. Gov't SO - Biochim Biophys Acta 1986 Dec 19;889(3):277-86 4 UI - 87076629 AU - Wright SE ; White JC TI - Membrane ordering effects of the anticancer agent VM-26. AB - The effect of the anticancer agent VM-26 on acyl chain order of cellular and model membranes was examined by electron spin resonance techniques. The order parameter for the paramagnetic probe 5-doxyl stearate was increased when VM-26 was incorporated into the bilayer of fluid-phase dimyristoylphosphatidylcholine (DMPC) or gel-phase dipalmitoylphosphatidylcholine (DPPC) liposomes at concentrations up to 4.8 mol%. The ordering effect of VM-26 in DMPC was greater than that of cholesterol on an equimolar basis. The less cytotoxic congener of VM-26, VP-16, was only one-third as active as VM-26 in its ordering effects on DMPC. Higher order parameters for 5-doxyl stearate were also noted in asolectin liposomes, Ehrlich ascites tumor cells, and CCRF-CEM cells treated with VM-26. We conclude that VM-26 has significant membrane associated activity in addition to its previously recognized nuclear effects. MH - Animal ; Carcinoma, Ehrlich Tumor/ULTRASTRUCTURE ; Cell Line ; Cell Membrane/*DRUG EFFECTS ; Cholesterol/PHARMACODYNAMICS ; Comparative Study ; Cyclic N-Oxides ; Dimyristoylphosphatidylcholine ; Electron Spin Resonance ; Etoposide/PHARMACODYNAMICS ; Human ; Leukemia/PATHOLOGY ; *Lipid Bilayers ; Liposomes ; Male ; Membrane Lipids ; Mice ; Podophyllotoxin/*ANALOGS & DERIVATIVES ; Spin Labels ; Support, Non-U.S. Gov't ; Teniposide/*PHARMACODYNAMICS ; 1,2-Dipalmitoylphosphatidylcholine SO - Biochim Biophys Acta 1986 Dec 16;863(2):297-304 1 UI - 87121292 AU - Itoh T ; Kawakami M ; Yamauchi Y ; Shimizu S ; Nakamura M TI - Effect of allopurinol on ischemia and reperfusion-induced cerebral injury in spontaneously hypertensive rats. AB - In spontaneously hypertensive rats, we studied the participation of xanthine oxidase-linked free radical in ischemia and reperfusion-induced cerebral injury, using allopurinol, a xanthine oxidase inhibitor. The loss of righting reflex was noted in some animals after a 4 hour occlusion of bilateral common carotid arteries and 19 of 25 animals died within 72 hours after reperfusion. One hour after reperfusion, the cerebral water content increased significantly, with an increase in sodium content and a decrease in potassium content. In 7 animals treated with oral administrations of allopurinol (200 mg/kg) 24 hours and 1 hour before occlusion, no death was found either during occlusion or after reperfusion, and the loss of righting reflex was noted in only one animal 24-72 hours following reperfusion. The increase in cerebral water content and accompanied changes in electrolyte contents were clearly prevented by allopurinol. These results suggest the possibility that the production of xanthine oxidase-linked free radical participates in cerebral injury due to ischemia and reperfusion in spontaneously hypertensive rats. MH - Allopurinol/*PHARMACODYNAMICS ; Animal ; Brain/DRUG EFFECTS/ METABOLISM ; Cerebral Ischemia/*PREVENTION & CONTROL/ PHYSIOPATHOLOGY ; Cerebrovascular Circulation ; Free Radicals ; Male ; Premedication ; Rats ; Rats, Inbred SHR ; Reflex/DRUG EFFECTS ; Superoxide/METABOLISM ; Xanthine Oxidase/*ANTAGONISTS & INHIBITORS/PHYSIOLOGY SO - Stroke 1986 Nov-Dec;17(6):1284-7 2 UI - 87076629 AU - Wright SE ; White JC TI - Membrane ordering effects of the anticancer agent VM-26. AB - The effect of the anticancer agent VM-26 on acyl chain order of cellular and model membranes was examined by electron spin resonance techniques. The order parameter for the paramagnetic probe 5-doxyl stearate was increased when VM-26 was incorporated into the bilayer of fluid-phase dimyristoylphosphatidylcholine (DMPC) or gel-phase dipalmitoylphosphatidylcholine (DPPC) liposomes at concentrations up to 4.8 mol%. The ordering effect of VM-26 in DMPC was greater than that of cholesterol on an equimolar basis. The less cytotoxic congener of VM-26, VP-16, was only one-third as active as VM-26 in its ordering effects on DMPC. Higher order parameters for 5-doxyl stearate were also noted in asolectin liposomes, Ehrlich ascites tumor cells, and CCRF-CEM cells treated with VM-26. We conclude that VM-26 has significant membrane associated activity in addition to its previously recognized nuclear effects. MH - Animal ; Carcinoma, Ehrlich Tumor/ULTRASTRUCTURE ; Cell Line ; Cell Membrane/*DRUG EFFECTS ; Cholesterol/PHARMACODYNAMICS ; Comparative Study ; Cyclic N-Oxides ; Dimyristoylphosphatidylcholine ; Electron Spin Resonance ; Etoposide/PHARMACODYNAMICS ; Human ; Leukemia/PATHOLOGY ; *Lipid Bilayers ; Liposomes ; Male ; Membrane Lipids ; Mice ; Podophyllotoxin/*ANALOGS & DERIVATIVES ; Spin Labels ; Support, Non-U.S. Gov't ; Teniposide/*PHARMACODYNAMICS ; 1,2-Dipalmitoylphosphatidylcholine SO - Biochim Biophys Acta 1986 Dec 16;863(2):297-304 3 UI - 87073796 AU - Jackson CV ; Mickelson JK ; Pope TK ; Rao PS ; Lucchesi BR TI - O2 free radical-mediated myocardial and vascular dysfunction. AB - In the present investigation electrolysis of a physiological buffer solution for 2 min with a constant current (20 mA, DC stainless steel anode) was observed to generate free radicals, determined by a luminol assay. Rabbit isolated hearts perfused with physiological buffer subjected to electrolysis were observed to undergo an increase in coronary artery perfusion pressure (PP) and in left ventricular end-diastolic pressure (LVEDP), 80 +/- 4 and 52 +/- 7 mmHg, respectively. Immediately after electrolysis of the physiological buffer, the hearts were observed to accumulate and retain (8-fold) more 125I-labeled albumin than hearts perfused with normal buffer without electrolysis, indicating an increased vascular permeability. The free radical scavengers, dimethyl sulfoxide (DMSO) and catalase (CAT), provided significant protection of the hearts against the changes in PP, LVEDP, and vascular permeability. This study demonstrates that toxic oxygen species generated independently of circulating blood elements or enzymatic reactions can have a direct effect on the vasculature of an isolated heart leading to alterations in cardiac function. The protection afforded by the addition of DMSO or CAT to the perfusion system would suggest that the OH. radical and H2O2 were the reactive oxygen species involved in producing the observed vascular and cardiac effects. MH - Animal ; Blood Pressure/DRUG EFFECTS ; Catalase/PHARMACODYNAMICS ; Coronary Vessels/DRUG EFFECTS/PATHOLOGY/*PHYSIOPATHOLOGY ; Dimethyl Sulfoxide/PHARMACODYNAMICS ; Electrolysis/ INSTRUMENTATION/METHODS ; Free Radicals ; Heart/DRUG EFFECTS/ *PHYSIOPATHOLOGY ; In Vitro ; Kinetics ; Male ; Myocardium/ PATHOLOGY ; Oxygen/*TOXICITY ; Perfusion ; Rabbits ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Am J Physiol 1986 Dec;251(6 Pt 2):H1225-31 4 UI - 87050276 AU - Angel MF ; Narayanan K ; Swartz WM ; Ramasastry SS ; Kuhns DB ; Basford RE ; Futrell JW TI - Deferoxamine increases skin flap survival: additional evidence of free radical involvement in ischaemic flap surgery. AB - This study presents further evidence of free radical involvement in skin flap necrosis in a dorsal rat flap model. Rats receiving deferoxamine, a free radical scavenger and iron chelator had significantly less necrosis (p less than 0.001) than saline treated rats. In a separate experiment, tissue determinations for malonyldialdehyde (MDA) were consistent with the survival results in showing a significant decrease in MDA in all biopsy sites (p less than 0.05 or less), indicative of reduced lipoperoxidation in the deferoxamine treated rats. MH - Animal ; Deferoxamine/*PHARMACODYNAMICS ; Free Radicals ; Graft Survival/*DRUG EFFECTS ; Male ; Malondialdehyde/METABOLISM ; Necrosis ; Rats ; Rats, Inbred Strains ; Skin/METABOLISM/ PATHOLOGY ; *Surgical Flaps ; Xanthine Oxidase/METABOLISM SO - Br J Plast Surg 1986 Oct;39(4):469-72 5 UI - 87033609 AU - Malis CD ; Bonventre JV TI - Mechanism of calcium potentiation of oxygen free radical injury to renal mitochondria. A model for post-ischemic and toxic mitochondrial damage. AB - With a variety of forms of ischemic and toxic tissue injury, cellular accumulation of Ca2+ and generation of oxygen free radicals may have adverse effects upon cellular and, in particular, mitochondrial membranes. Damage to mitochondria, resulting in impaired ATP synthesis and diminished activity of cellular energy-dependent processes, could contribute to cell death. In order to model, in vitro, conditions present post-ischemia or during toxin exposure, the interactions between Ca2+ and oxygen free radicals on isolated renal mitochondria were characterized. The oxygen free radicals were generated by hypoxanthine and xanthine oxidase to simulate in vitro one of the sources of oxygen free radicals in the early post-ischemic period in vivo. With site I substrates, pyruvate and malate, Ca2+ pretreatment, followed by exposure to oxygen free radicals, resulted in an inhibition of electron transport chain function and complete uncoupling of oxidative phosphorylation. These effects were partially mitigated by dibucaine, a phospholipase A2 inhibitor. With the site II substrate, succinate, the electron transport chain defect was not manifest and respiration remained partially coupled. The electron transport chain defect produced by Ca2+ and oxygen free radicals was localized to NADH CoQ reductase. Calcium and oxygen free radicals reduced mitochondrial ATPase activity by 55% and adenine nucleotide translocase activity by 65%. By contrast oxygen free radicals alone reduced ATPase activity by 32% and had no deleterious effects on translocase activity. Dibucaine partially prevented the Ca2+-dependent reduction in ATPase activity and totally prevented the Ca2+-dependent translocase damage observed in the presence of oxygen free radicals. These findings indicate that calcium potentiates oxygen free radical injury to mitochondria. The Ca2+-induced potentiation of oxygen free radical injury likely is due in part to activation of phospholipase A2. This detrimental interaction associated with Ca2+ uptake by mitochondria and exposure of the mitochondria to oxygen free radicals may explain the enhanced cellular injury observed during post-ischemic reperfusion. MH - Animal ; Calcium/*PHARMACODYNAMICS ; Dibucaine/PHARMACODYNAMICS ; Free Radicals ; H(+)-Transporting ATPase/METABOLISM ; Ischemia/ PATHOLOGY ; Kidney/BLOOD SUPPLY/DRUG EFFECTS/*ULTRASTRUCTURE ; Malates/METABOLISM ; Male ; Mitochondria/DRUG EFFECTS/*METABOLISM ; Models, Biological ; Multienzyme Complexes/METABOLISM ; Oligomycins/PHARMACODYNAMICS ; Oxygen Consumption ; Oxygen/ *TOXICITY ; Phosphotransferases, ATP/METABOLISM ; Pyruvates/ METABOLISM ; Quinone Reductases/METABOLISM ; Rats ; Rats, Inbred Strains ; Support, U.S. Gov't, P.H.S. SO - J Biol Chem 1986 Oct 25;261(30):14201-8 6 UI - 87028677 AU - Menasche P ; Grousset C ; Gauduel Y ; Mouas C ; Piwnica A TI - Enhancement of cardioplegic protection with the free-radical scavenger peroxidase. AB - This study assesses the ability of the free-radical scavenger peroxidase to enhance cardioplegic protection when given during or before myocardial ischemia. Forty-four isolated isovolumetric buffer-perfused rat hearts were studied. In a first series of experiments that consisted of three groups, hearts were subjected to 90 min of normothermic global ischemia followed by 45 min of reperfusion. One group received a crystalloid cardioplegic solution given as a single dose at the onset of arrest. A second group received cardioplegic solution supplemented with superoxide dismutase (200,000 U/liter), and a third group received cardioplegic solution supplemented with peroxidase (6000 U/liter). Based on comparisons of postreperfusion coronary flow, left ventricular developed pressure, maximum dP/dt, and diastolic pressure, we found that the best protection was provided by peroxidase-enriched cardioplegia. A second series of experiments was then undertaken to assess the effects of the latter enzyme given as a pretreatment. Hearts were subjected to 3 hr of global ischemia, during which myocardial protection was provided by hypothermia (15 degrees C) along with multidose cardioplegia. The treatment group was given peroxidase (10,000 U/liter) added to the perfusate fluid for 15 min before the onset of cardioplegic arrest without further enzyme supplementation during ischemia or reperfusion. Hearts perfused with standard buffer for an equal period of time served as controls. While the two groups demonstrated the same degree of postischemic increase in myocardial stiffness, peroxidase-pretreated hearts had a significantly better recovery of contractile indexes at 30 and 45 min of reflow.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Animal ; Compliance ; Coronary Circulation ; Free Radicals ; *Heart Arrest, Induced ; Heart Function Tests ; Heart Ventricle ; Heart/PHYSIOLOGY ; Hydrogen Peroxide/*ANTAGONISTS & INHIBITORS ; In Vitro ; Isoenzymes/*THERAPEUTIC USE ; Myocardial Diseases/ PREVENTION & CONTROL ; Myocardium/PATHOLOGY ; Peroxidases/ *THERAPEUTIC USE ; Rats ; Rats, Inbred Strains SO - Circulation 1986 Nov;74(5 Pt 2):III138-44 7 UI - 87023373 AU - Grisham MB ; Hernandez LA ; Granger DN TI - Xanthine oxidase and neutrophil infiltration in intestinal ischemia. AB - A growing body of experimental data indicates that reactive oxygen metabolites such as superoxide, hydrogen peroxide, and hydroxyl radical may mediate the mucosal injury produced by reperfusion of ischemic intestine. Xanthine oxidase has been proposed as the primary source of these reduced O2 species because pretreatment with xanthine oxidase inhibitors such as allopurinol or pterin aldehyde prevent postischemic mucosal injury. Another potential source of oxygen radicals is the inflammatory neutrophil. To ascertain whether neutrophils could play a role in the pathogenesis of ischemia-reperfusion injury in the small bowel we examined the effect of ischemia and reperfusion on neutrophil infiltration and tissue levels of reduced glutathione, superoxide dismutase, and catalase. Our studies demonstrate that reperfusion of ischemic intestines results in a dramatic increase (1,800%) in neutrophil infiltration and a concurrent loss of reduced glutathione and superoxide dismutase of 60 and 30%, respectively. Catalase activity was unaffected by ischemia-reperfusion. Pretreatment with allopurinol or administration of superoxide dismutase prevented the influx of neutrophils and retarded the drop in reduced glutathione levels. These results suggest a relationship among xanthine oxidase-generated oxy radicals, neutrophil extravasation, and mucosal damage. We propose that ischemia and reperfusion results in xanthine oxidase-generated, superoxide-dependent accumulation of inflammatory neutrophils in the mucosa where neutrophil-derived reactive oxygen metabolites mediate and/or exacerbate intestinal injury. MH - Allopurinol/PHARMACODYNAMICS ; Animal ; Cats ; Free Radicals ; Glutathione/METABOLISM ; Intestinal Mucosa/ENZYMOLOGY/PATHOLOGY ; Intestines/*BLOOD SUPPLY ; Ischemia/*ENZYMOLOGY/PATHOLOGY ; Myeloperoxidase/METABOLISM ; Neutrophils/DRUG EFFECTS/ENZYMOLOGY/ *PATHOLOGY ; Oxygen/METABOLISM ; Superoxide Dismutase/METABOLISM/ PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Xanthine Oxidase/*METABOLISM SO - Am J Physiol 1986 Oct;251(4 Pt 1):G567-74 8 UI - 87013189 AU - Otani H ; Umemoto M ; Kagawa K ; Nakamura Y ; Omoto K ; Tanaka K ; Sato T ; Nonoyama A ; Kagawa T TI - Protection against oxygen-induced reperfusion injury of the isolated canine heart by superoxide dismutase and catalase. AB - While oxygen-derived free radicals have been implicated in the pathogenesis of myocardial injury, the exact nature of this injury is still unclear. To test the hypothesis that oxygen-induced injury may influence the recovery of cardiac function from ischemic damage, we used an oxygen free radical scavenger, superoxide dismutase (SOD), together with catalase, during the reperfusion of isolated canine heart which had been subjected to 15 min of normothermic ischemic arrest followed by 2 hr of hypothermic cardioplegic preservation using a modified Collins solution. Determinations of thiobarbituric acid reactive substances and coenzyme Q10 within the myocardium showed that the treatment with SOD and catalase was capable of inhibiting lipid peroxidation induced by reperfusion. This inhibition was apparently associated with the improvement of myocardial energy metabolism and cardiac performance. Coronary flow was significantly higher in the heart treated with SOD and catalase during the working stage with a corresponding increase in oxygen consumption. Myocardial adenosine triphosphate (ATP) was partially, but significantly restored during reperfusion in these hearts whereas no restoration was observed in the heart without the enzymes. The treatment with SOD and catalase also improved left ventricular stroke work index and left ventricular maximum dp/dt at an early stage of the working mode. These results suggest that the use of SOD and catalase during reperfusion can protect the ischemic heart against reperfusion injury by scavenging oxygen-derived free radicals. MH - Animal ; Catalase/*THERAPEUTIC USE ; *Coronary Circulation ; Coronary Disease/*PHYSIOPATHOLOGY ; Dogs ; Free Radicals ; Heart Arrest, Induced ; In Vitro ; *Oxygen ; Superoxide Dismutase/ *THERAPEUTIC USE ; Support, Non-U.S. Gov't SO - J Surg Res 1986 Aug;41(2):126-33 9 UI - 87003348 AU - Hansson R ; Johansson S ; Jonsson O ; Pettersson S ; Scherst:en T ; Waldenstr:om J TI - Kidney protection by pretreatment with free radical scavengers and allopurinol: renal function at recirculation after warm ischaemia in rabbits. AB - Renal function and morphology were studied before and after 60 min of renal ischaemia and contralateral nephrectomy in five groups of rabbits. The animals were pretreated with superoxide dismutase, catalase, allopurinol or mannitol. One group was not pretreated and served as a control. A moderate transient increase in serum creatinine concentration was observed in the control rabbits, while a significantly less pronounced increase was noted after pretreatment with superoxide dismutase, catalase and mannitol. Pretreatment with allopurinol did not significantly reduce the postoperative increase in serum creatinine and sodium excretion, but the urine osmolality returned to normal more rapidly than in the control group. The appearance under the light microscope of kidney tissue taken from surviving rabbits was found to be normal irrespective of pretreatment. Severe tubular necrosis was observed in the kidneys from rabbits that died during the observation period. MH - Allopurinol/*PHARMACODYNAMICS ; Animal ; Catalase/ PHARMACODYNAMICS ; Comparative Study ; Creatinine/BLOOD ; Free Radicals ; Ischemia/*PHYSIOPATHOLOGY ; Kidney/*BLOOD SUPPLY/DRUG EFFECTS/PHYSIOPATHOLOGY ; Mannitol/PHARMACODYNAMICS ; Nephrectomy ; Rabbits ; Superoxide Dismutase/PHARMACODYNAMICS ; Support, Non-U.S. Gov't SO - Clin Sci 1986 Sep;71(3):245-51 10 UI - 87002887 AU - Czerniecki B ; Gad SC ; Reilly C ; Smith AC ; Witz G TI - Phorbol diacetate inhibits superoxide anion radical production and tumor promotion by mezerein. AB - The ability of the non-promoter phorbol diacetate (PDA) to modulate superoxide anion radical production by the complete tumor promoter phorbol myristate acetate (PMA) or the second stage promoter mezerein was assessed. Superoxide anion radical production was measured by the superoxide dismutase inhibitable reduction of nitroblue tetrazolium (NBT) to a blue intracellular formazan precipitate. These studies demonstrated that superoxide anion radical production by murine peritoneal exudate cells (PEC) stimulated by i.p. injection with mezerein (100 ng) is inhibited in a dose-dependent manner by co-administration with PDA (1-1000 ng). There was no effect on the number of formazan-positive PEC when PDA was co-administered with PMA. In a two-stage tumor promotion bioassay in female SENCAR mice initiated with 25.6 micrograms dimethylbenz[a]anthracene (DMBA) followed by first stage promotion with PMA (4X, 2 micrograms), co-administration of mezerein (2 micrograms) with 2 micrograms or 20 micrograms PDA reduced the number of papillomas after 14 weeks by 38% and 44%, respectively, compared with mezerein treatment alone. PDA (20 micrograms) when co-administered with mezerein (2 micrograms) does not inhibit mezerein induced hyperplasia in mouse skin. These results suggest a correlation between the ability of PDA to inhibit both superoxide anion radical production and tumor promotion by mezerein. MH - Animal ; *Carcinogens ; Female ; Free Radicals ; Hyperplasia ; Macrophages/METABOLISM ; Mice ; Phorbol Esters/*PHARMACODYNAMICS ; Skin Neoplasms/CHEMICALLY INDUCED/*PREVENTION & CONTROL ; Skin/ PATHOLOGY ; Superoxide/*METABOLISM ; Support, U.S. Gov't, P.H.S. ; Terpenes/*TOXICITY ; Tetradecanoylphorbol Acetate SO - Carcinogenesis 1986 Oct;7(10):1637-41 11 UI - 86319279 AU - Hearse DJ ; Manning AS ; Downey JM ; Yellon DM TI - Xanthine oxidase: a critical mediator of myocardial injury during ischemia and reperfusion? AB - Myocardial ischemia initiates a series of cellular reactions which unless checked will culminate in cell death and tissue necrosis. Although reperfusion provides a means of preventing cell death it is not without hazard. In cases of mild ischemia, where tissue injury is in its reversible phase, reperfusion may precipitate potentially lethal ventricular arrhythmias and in cases of severe injury it may actually accelerate the process of cell death leading to hemorrhage and other forms of severe injury. The identity of mediators of cellular injury, and particularly the critical transition from reversible to irreversible injury, remains controversial. Whereas for a number of years ATP depletion, calcium overload and catecholamines have been considered as key factors in tissue injury, attention has recently been directed towards oxygen-derived free radicals (e.g. superoxide and the hydroxyl radical). In this article we discuss sources of free radicals in the mammalian heart (xanthine oxidase, mitochondria, leucocytes, and catecholamines) and present arguments based on quantitative and temporal considerations that the xanthine oxidase-mediated degradation of hypoxanthine is the most important source of free radicals and as such is the most appropriate target for therapeutic intervention. To support our arguments we present data from two species, the dog and the rat, in which we have shown how allopurinol, the specific inhibitor of xanthine oxidase, can afford a reduction of infarct size in the dog and can dramatically reduce the incidence of potentially lethal reperfusion-induced arrhythmias in the rat. Arising from these and other studies is the proposition that anti-free radical interventions (particularly those directed towards xanthine oxidase inhibition) may provide an important new therapeutic principle in the management of ischemia and reperfusion. MH - Allopurinol/PHARMACODYNAMICS ; Animal ; Arrhythmia/DRUG THERAPY/ METABOLISM/PHYSIOPATHOLOGY ; Catecholamines/METABOLISM ; Coronary Disease/DRUG THERAPY/ENZYMOLOGY/*PATHOLOGY/PHYSIOPATHOLOGY ; Free Radicals ; Human ; Leukocytes/ENZYMOLOGY ; Mitochondria, Heart/ METABOLISM ; Myocardial Infarction/DRUG THERAPY/ENZYMOLOGY/ *PATHOLOGY/PHYSIOPATHOLOGY ; Oxygen/METABOLISM ; Perfusion ; Review ; Support, Non-U.S. Gov't ; Xanthine Oxidase/*PHYSIOLOGY SO - Acta Physiol Scand [Suppl] 1986;548:65-78 12 UI - 86319275 AU - Sanfey H ; Sarr MG ; Bulkley GB ; Cameron JL TI - Oxygen-derived free radicals and acute pancreatitis: a review. AB - The role of oxygen-derived free radicals in the pathogenesis of acute pancreatitis has been investigated in a series of studies using an ex vivo, perfused canine pancreas preparation. Three models of experimental acute pancreatitis have been developed in this preparation: ischemic pancreatitis, gallstone pancreatitis, and alcohol-induced pancreatitis. In each model, the pancreas becomes edematous, gains weight, and the perfusate develops hyperamylasemia during the 4 hour period of perfusion. Pretreatment with the free radical scavengers superoxide dismutase and catalase significantly ameliorates these manifestations of pancreatic injury in each of the three models. The source of the free radical generation was investigated by pretreating the preparation with allopurinol, a quite specific inhibitor of xanthine oxidase. In each of the three models, this also significantly ameliorated the injury process. These experiments demonstrate that oxygen-derived free radicals, generated by activated xanthine oxidase, appear to play a central role in the pathogenesis of acute pancreatitis in these models. These findings shed light on the fundamental pathophysiology of this disease and may provide the basis for more effective therapy in the future. MH - Acute Disease ; Alcoholism/COMPLICATIONS/ENZYMOLOGY ; Allopurinol/ PHARMACODYNAMICS ; Animal ; Catalase/DIAGNOSTIC USE ; Disease Models, Animal ; Free Radicals ; Human ; Ischemia/ENZYMOLOGY/ ETIOLOGY ; Oxygen/*TOXICITY ; Pancreas/BLOOD SUPPLY ; Pancreatitis/*CHEMICALLY INDUCED/ENZYMOLOGY/PATHOLOGY ; Review ; Superoxide Dismutase/DIAGNOSTIC USE ; Support, U.S. Gov't, P.H.S. ; Xanthine Oxidase/ANTAGONISTS & INHIBITORS SO - Acta Physiol Scand [Suppl] 1986;548:109-18 13 UI - 86298163 AU - Shinkai K ; Mukai M ; Akedo H TI - Superoxide radical potentiates invasive capacity of rat ascites hepatoma cells in vitro. AB - Effect of superoxide radical (O2-) produced extracellularly by hypoxanthine (HX) and xanthine oxidase (XO) on invasive capacity of rat ascites hepatoma cells was studied. Invasive capacity was estimated in vitro by counting the number of tumor cell colonies penetrated underneath cultured mesothelial cell monolayer. When the tumor cells had been treated with non-toxic doses of HX and XO, the formation of penetrated colonies increased with increasing concentrations of XO. This increment was completely inhibited by scavengers of active oxygen radicals, superoxide dismutase (SOD) in combination with catalase (CAT) added simultaneously at the time of HX-XO treatment. MH - Animal ; Catalase/PHARMACODYNAMICS ; Cells, Cultured ; Free Radicals ; Hypoxanthines/PHARMACODYNAMICS ; Liver Neoplasms, Experimental/*PATHOLOGY ; Macrophages/PHYSIOLOGY ; Neoplasm Invasiveness ; Superoxide/*TOXICITY ; Superoxide Dismutase/ PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Xanthine Oxidase/ PHARMACODYNAMICS SO - Cancer Lett 1986 Jul;32(1):7-13 14 UI - 86294769 AU - Woodruff T ; Blake DR ; Freeman J ; Andrews FJ ; Salt P ; Lunec J TI - Is chronic synovitis an example of reperfusion injury? AB - In an attempt to define why the joint synovial cavity is prone to develop persistent synovial inflammation we show that hypoxia is induced by pressure changes caused by exercise in the presence of an inflammatory effusion. On resting 'reperfusion injury' may take place. The biochemistry of reperfusion injury has only recently been defined and perhaps surprisingly for an insult that has hypoxia as its central ingredient involves the subsequent production of oxygen derived free radical species. We apply the reaction sequences that are believed to occur during hypoxic/reperfusion injury to the joint synovial cavity and, on the basis of reported 'in vivo' observations, suggest novel therapeutic approaches that we believe are applicable to the treatment of persistent synovial inflammation. MH - Animal ; Anoxia/COMPLICATIONS ; Cats ; Chronic Disease ; Exertion ; Free Radicals ; Human ; Ischemia/ETIOLOGY ; Knee Joint/ PHYSIOPATHOLOGY ; Oxygen/METABOLISM ; Partial Pressure ; Perfusion ; Rats ; Synovitis/*PHYSIOPATHOLOGY ; Xanthine Oxidase/ METABOLISM ; Xanthines/METABOLISM SO - Ann Rheum Dis 1986 Jul;45(7):608-11 15 UI - 86056804 AU - Granger DN ; McCord JM ; Parks DA ; Hollwarth ME TI - Xanthine oxidase inhibitors attenuate ischemia-induced vascular permeability changes in the cat intestine. AB - Previous reports indicate that allopurinol, a xanthine oxidase inhibitor, largely prevents the injury produced by reperfusion of ischemic tissues. In order to further assess the role of xanthine oxidase in ischemia-reperfusion injury, we examined the influence of another inhibitor of the enzyme (pterin aldehyde) on the increased vascular permeability produced by intestinal ischemia. Vascular permeability estimates in autoperfused segments of cat ileum were derived from the relationship between lymph-to-plasma protein concentration ratio and lymph flow. One hour of intestinal ischemia increased vascular permeability to 0.43 +/- 0.02 from a control (nonischemic) value of 0.08 +/- 0.005. In ischemic ileal segments pretreated with purified pterin aldehyde, vascular permeability increased to only 0.15 +/- 0.02. Pretreatment with commercially prepared folic acid, which is contaminated with pterin aldehyde, also attenuated the ischemia-induced increase in vascular permeability (0.16 +/- 0.04). These findings support the hypothesis that xanthine oxidase is a major source of oxygen-free radicals produced during reperfusion of the ischemic small bowel. MH - Animal ; Blood Proteins/ANALYSIS ; Capillary Permeability/*DRUG EFFECTS ; Cats ; Folic Acid/PHARMACODYNAMICS ; Free Radicals ; Ileum/*BLOOD SUPPLY ; Ischemia/*PHYSIOPATHOLOGY ; Lymph/ANALYSIS/ SECRETION ; Pteridines/*PHARMACODYNAMICS ; Support, U.S. Gov't, P.H.S. ; Time Factors ; Xanthine Oxidase/*ANTAGONISTS & INHIBITORS/PHYSIOLOGY SO - Gastroenterology 1986 Jan;90(1):80-4 16 UI - 86322649 AU - Rao NA ; Romero JL ; Fernandez MA ; Sevanian A ; Marak GE Jr TI - Effect of iron chelation on severity of ocular inflammation in an animal model. AB - Metabolites of oxygen-free radicals generated by polymorphonuclear leukocytes and macrophages are believed to inflict the initial tissue damage in acute inflammations. Of the various oxygen products, hydroxyl radicals are known to be potent toxic agents, and their generation depends largely on the presence of free iron. Treatment of experimental uveitis in Lewis rats with an iron chelator, deferoxamine mesylate, resulted in marked reduction in choroidal inflammation and suppression of retinal damage. These findings suggest that in experimental uveitis the severity of ocular inflammation and tissue damage may be mediated by the iron-catalyzed generation of hydroxyl radicals, and deferoxamine may thus serve as an anti-inflammatory agent. MH - Animal ; *Anti-Inflammatory Agents ; Antigens ; Choroiditis/ PATHOLOGY ; Deferoxamine/*PHARMACODYNAMICS/THERAPEUTIC USE ; Disease Models, Animal ; Free Radicals ; Hydroxides ; Rats ; Rats, Inbred LEW ; Retinitis/PATHOLOGY ; Support, U.S. Gov't, P.H.S. ; Uveitis/*DRUG THERAPY/ETIOLOGY/PATHOLOGY SO - Arch Ophthalmol 1986 Sep;104(9):1369-71 17 UI - 86319276 AU - Martin D ; Korthuis RJ ; Perry M ; Townsley MI ; Taylor AE TI - Oxygen radical-mediated lung damage associated with alpha-naphthylthiourea. AB - Lungs were damaged with alpha-naphthylthiourea (ANTU) and various compounds were used to block its effect. Although the results are variable, superoxide dismutase, catalase and dimethylsulfoxide all protected against ANTU, indicating that OH radicals are responsible for this type of lung injury. Leukocytes do not appear to be required for the damage to occur; however, hydroxurea (given over 2 days) did block the ANTU damage when neutrophils were decreased to 1/2 normal values or when administered acutely. The free radicals may be generated by the cyclooxygenase pathway since ibuprofen blocked the ANTU damage, whereas blocking xanthine oxidase using allopurinol failed to prevent the lung damage. MH - Animal ; Blood Pressure/DRUG EFFECTS ; Capillary Permeability/ DRUG EFFECTS ; Dimethyl Sulfoxide/PHARMACODYNAMICS ; Disease Models, Animal ; Dogs ; Endothelium/METABOLISM/PATHOLOGY/ SECRETION ; Free Radicals ; Lung/DRUG EFFECTS/*PATHOLOGY/ PHYSIOPATHOLOGY ; Oxygen/*TOXICITY ; Pulmonary Circulation/DRUG EFFECTS ; Superoxide Dismutase/METABOLISM/THERAPEUTIC USE ; Support, Non-U.S. Gov't ; Thiourea/*ANALOGS & DERIVATIVES/ TOXICITY SO - Acta Physiol Scand [Suppl] 1986;548:119-25 18 UI - 86308888 AU - Paller MS TI - Hypothyroidism protects against free radical damage in ischemic acute renal failure. AB - The effect of hypothyroidism on ischemic acute renal failure was studied in rats. Ten days after thyroidectomy with parathyroid reimplantation, rats underwent right uninephrectomy followed by occlusion of the left renal artery for 60 min. Plasma creatinine was lower in thyroidectomized than control rats 24 hr after ischemia; 1.3 +/- 0.5 vs. 3.2 +/- 0.6 mg%; P less than 0.05. Twenty-four hours after ischemia, inulin clearance was higher in thyroidectomized than control animals (0.40 +/- 0.06 vs. 0.17 +/- 0.03 mliter/min; P less than 0.01), despite an initially lower inulin clearance in thyroidectomized animals (0.81 vs. 1.1 +/- 0.07 mliter/min; P less than 0.05). Administration of the antithyroid drug prophylthiouracil for 14 days also resulted in lower plasma creatinine after ischemia. Kidneys from thyroidectomized animals showed less histologic damage 24 hr after ischemia. Renal cortical content of the lipid peroxidation product malondialdehyde was increased less in thyroidectomy than control kidneys after 60 min ischemia plus 15 min reflow (0.08 +/- 0.02 vs. 0.42 +/- 0.1 nmole/mg protein; P less than 0.005). Renal cortical glutathione content was higher in thyroidectomized animals by approximately 36%, 650 +/- 46 vs. 479 +/- 32 nmole/mg protein (P less than 0.02). In normal rats, glutathione infusion also increased renal cortical glutathione content and resulted in lower plasma creatinine 24 hr after renal artery ischemia. Therefore, hypothyroidism resulted in functional and histologic protection against injury after ischemia. Post-ischemic renal lipid peroxidation was reduced in thyroidectomized animals, perhaps the result of increased scavenging of reactive oxygen species (oxygen free radicals and H2O2) by glutathione. MH - Animal ; Free Radicals ; Glutathione/PHYSIOLOGY ; Hypothyroidism/ *COMPLICATIONS ; Ischemia/*PHYSIOPATHOLOGY ; Kidney/*BLOOD SUPPLY/ PHYSIOLOGY ; Kidney Failure, Acute/*COMPLICATIONS/METABOLISM/ PATHOLOGY ; Lipid Peroxides/METABOLISM ; Male ; Propylthiouracil/ PHARMACODYNAMICS ; Rats ; Support, U.S. Gov't, P.H.S. SO - Kidney Int 1986 Jun;29(6):1162-6 19 UI - 86298907 AU - McKechnie K ; Furman BL ; Parratt JR TI - Modification by oxygen free radical scavengers of the metabolic and cardiovascular effects of endotoxin infusion in conscious rats. AB - Since oxygen free radicals may have a role in the pathophysiology of endotoxin shock, we have studied the effects of a wide range of compounds (alpha-tocopherol, reduced glutathione, allopurinol, superoxide dismutase (alone or in combination with catalase) and phenyl butylnitrone) that can act either to remove free radicals as they are generated or to prevent their generation. The effects of these substances on the metabolic and cardiovascular responses to endotoxin were examined in conscious rats. The intravenous infusion of endotoxin (10 mg/kg i.v. given over 4 h) resulted in systemic hypotension, transient tachycardia, an increase in plasma lactate, and an initial hyperglycemia followed, in those rats that died before 24 h, by hypoglycemia. The hypotension and tachycardia produced by endotoxin were not significantly modified by alpha-tocopherol, allopurinol, or superoxide dismutase, alone or in combination with catalase. The tachycardia was attenuated by reduced glutathione and phenyl butylnitrone. alpha-Tocopherol attenuated the initial hyperglycemia produced by endotoxin whilst alpha-tocopherol, allopurinol, and phenyl butylnitrone all significantly attenuated the endotoxin-induced increase in plasma lactate. Among the free radical scavenging systems studied, only alpha-tocopherol and phenyl butylnitrone improved survival. These results suggest a contribution from oxygen-free radicals to the pathophysiology of endotoxemia. MH - Allopurinol/PHARMACODYNAMICS ; Animal ; Blood Glucose/METABOLISM ; Cardiovascular System/*DRUG EFFECTS/PHYSIOLOGY ; Catalase/ PHARMACODYNAMICS ; Endotoxins/*TOXICITY ; Free Radicals ; Glutathione/PHARMACODYNAMICS ; Lactates/BLOOD ; Male ; Nitrogen Oxides/PHARMACODYNAMICS ; Oxygen/*METABOLISM ; Prostaglandins/ BLOOD ; Rats ; Rats, Inbred Strains ; Shock, Septic/ETIOLOGY/ PHYSIOPATHOLOGY ; Superoxide Dismutase/PHARMACODYNAMICS ; Vitamin E/BLOOD/PHARMACODYNAMICS SO - Circ Shock 1986;19(4):429-39 20 UI - 86277740 AU - Jin LJ ; Lalonde C ; Demling RH TI - Lung dysfunction after thermal injury in relation to prostanoid and oxygen radical release. AB - We studied whether changes in lung function after burns (1- to 12-h period) were due to changes in lung water or airways resistance and the relationship of the changes to prostanoid and O2 radical activity (measured as lipid peroxidation). Twenty-five anesthetized mechanically ventilated adult sheep were given a 40% of body surface scald burn and resuscitated to restore and maintain base-line filling pressures. Dynamic lung compliance (Cdyn) decreased by 40% from 38 +/- 5 to 24 +/- 4 ml/cmH2O at 12 h. Venous thromboxane B2 transiently increased from 210 +/- 40 to 1,100 +/- 210 pg/ml, and the value in lung lymph increased from 180 +/- 80 to 520 +/- 80 pg/ml. Prostacyclin levels in lung lymph and plasma remained at base line. Protein-poor lung lymph flow increased two- to threefold, but postmortem lung analysis revealed no increase in lung water from the control of 3.5 +/- 0.3 g H2O/g dry wt. No increase in protein permeability was seen. However, the lipid peroxidation of lung tissue measured as malondialdehyde was significantly increased from the control value of 56 +/- 4 nmol/g lung to a value of 69 +/- 6. Ibuprofen pretreatment (12.5 mg/kg) markedly attenuated the decrease in Cdyn, with the value at 12 h being 90% of base line. Ibuprofen also decreased the amount of lung lipid peroxidation but did not decrease the lung lymph response. We conclude that the decrease in Cdyn seen early postburn is not due to increased lung water, but, rather, is due to a mediator-induced bronchoconstriction, attenuated by ibuprofen; the mediator being either thromboxane or a byproduct of O2 radicals as evidenced by increased lipid peroxide production in lung tissue. MH - Anesthesia ; Animal ; Burns/DRUG THERAPY/METABOLISM/ *PHYSIOPATHOLOGY ; Free Radicals ; Ibuprofen/THERAPEUTIC USE ; Lung/METABOLISM/*PHYSIOPATHOLOGY ; Oxygen/*METABOLISM ; Postmortem Changes ; Prostaglandins/*METABOLISM ; Respiratory Function Tests ; Sheep ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - J Appl Physiol 1986 Jul;61(1):103-12 21 UI - 86265778 AU - Warren JS ; Ward PA TI - Oxidative injury to the vascular endothelium. AB - In recent years, increasing clinical and experimental data have provided compelling evidence that neutrophil-derived oxygen radicals and their metabolites are important mediators of vascular endothelial injury. In this review, attention is directed toward in vitro, ex vivo, and in vivo experimental studies contributing to current understanding of mechanisms of oxyradical mediated endothelial damage. Although these studies may have broad significance, they appear to have particular relevance to the pathogenesis of the adult respiratory distress syndrome and other types of acute pulmonary dysfunction. MH - Animal ; Blood Vessels/*METABOLISM/PATHOLOGY ; Endothelium/ METABOLISM/PATHOLOGY ; Free Radicals ; Lung/PATHOLOGY ; Microscopy, Electron ; Neutrophils/METABOLISM ; Oxygen/ *METABOLISM ; Peroxides/METABOLISM ; Rabbits ; Rats ; Superoxide Dismutase/METABOLISM ; Umbilical Veins/METABOLISM/PATHOLOGY SO - Am J Med Sci 1986 Aug;292(2):97-103 22 UI - 86260256 AU - Hladovec J TI - Protective effect of oxygen-derived free radical scavengers on the endothelium in vivo. AB - The endothelo-protective activity of a series of low-molecular oxygen-derived free radical scavengers (OFRS) was tested in rats. A model of endothelaemia provoked by intravenous administration of hydrogen peroxide was used. With each OFRS the activity in the hydrogen peroxide model was compared with that in the less specific model using the provocation by citrate as a calcium chelating agent. Relatively unspecific but biologically important OFRS, ascorbic acid, tocopherol, troxerutin and glutathione were tested in the first phase of the study. A marked optimum of endothelo-protective activity was shown with all agents, the optimum against hydrogen peroxide having been observed at doses from 3 to 50 times lower than against citrate. Ascorbic acid, troxerutin and the combination of both were also tested in another model based on leg ischaemia produced by ligature of the common femoral artery. Without OFRS, a marked increase of endothelaemia was observed after 30-60 min ischaemia showing a second peak after the release of the ligature. This second peak was completely abolished by the preventive administration of OFRS in a dose which was also effective in the hydrogen peroxide model. MH - Animal ; Anticoagulants/*PHARMACODYNAMICS ; Ascorbic Acid/ *PHARMACODYNAMICS ; Aspirin/*PHARMACODYNAMICS ; Coronary Disease/ *PHYSIOPATHOLOGY ; Endothelium/DRUG EFFECTS/*PHYSIOLOGY ; Female ; Femoral Artery/PHYSIOLOGY ; Free Radicals ; Heparin/ *PHARMACODYNAMICS ; Hydrogen Peroxide/*TOXICITY ; Hydroxyethylrutoside/*ANALOGS & DERIVATIVES/PHARMACODYNAMICS ; Kinetics ; Lactates/PHARMACODYNAMICS ; Perfusion ; Pyruvates/ PHARMACODYNAMICS ; Rats ; Rats, Inbred Strains ; Rutin/*ANALOGS & DERIVATIVES ; Vitamin E/*ANALOGS & DERIVATIVES/PHARMACODYNAMICS SO - Physiol Bohemoslov 1986;35(2):97-103 23 UI - 86255633 AU - Jackson CV ; Mickelson JK ; Stringer K ; Rao PS ; Lucchesi BR TI - Electrolysis-induced myocardial dysfunction. A novel method for the study of free radical mediated tissue injury. AB - Oxygen-derived free radicals and other oxidizing species are thought to be involved in inflammation and ischemic tissue injuries. Recently, oxygen-derived free radicals also have been implicated in tissue injury of the myocardium subjected to ischemia/reperfusion. The purpose of this investigation was to determine if electrolysis of a physiological buffer would serve as a source of free radicals, and if these radicals would lead to alterations in myocardial function. Isolated Langendorff-perfused rabbit hearts perfused with buffer subjected to a 20 mA D.C. current for 2 min demonstrated significant increases in coronary perfusion pressure (37 +/- 6 mmHg), left ventricular end diastolic pressure (41 +/- 7 mmHg), and loss in left ventricular developed pressure (35 +/- 5%). The free radical scavengers, superoxide dismutase and a combination of tryptophan plus glycine, were effective in protecting the hearts from the effects of electrolysis. The presence of free radicals was semiquantitated with a radical-luminol chemiluminescent assay. In this assay a variety of radical scavengers and antioxidants were effective (i.e., dimethyl sulfoxide, nitro blue tetrazolium, ascorbate, superoxide dismutase, 1, 3-diphenylisobenzofuran, and glycine, catalase), whereas mannitol and tryptophan were not effective. The data indicate that electrolysis of a physiological buffer produces a milieu containing several reactive oxygen species or free radicals that have the potential to produce alterations in a biological system. This method has the advantage over existing protocols for the generation of radicals in that it is a blood-free and an enzyme-free system. MH - Animal ; Buffers ; Catalase/PHARMACODYNAMICS ; Electric Stimulation ; Electrolysis ; Free Radicals ; Heart/DRUG EFFECTS/ *PHYSIOPATHOLOGY ; Hydrogen Peroxide/PHARMACODYNAMICS ; Male ; Methods ; Myocardium/*PATHOLOGY ; Rabbits ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - J Pharmacol Methods 1986 Jul;15(4):305-20 24 UI - 86245799 AU - Balin AK ; Allen RG TI - Mechanisms of biologic aging. AB - The authors outline the progression of thought on the mechanism of the aging process, giving emphasis to environmental factors that influence genetic events. Discussion is limited to those theories that explain fundamental causes of aging and have a firm thermodynamic basis. The authors propose that the cumulative result of continual oxidative stress and other thermodynamic processes (such as amino-acid racemization and nonenzymatic glycosylation), resulting in altered function and increasing the net entropy of living systems, governs the rate of the aging process. MH - *Aging ; Amino Acids/METABOLISM ; Animal ; Antioxidants/ METABOLISM ; Catalase/METABOLISM ; Energy Metabolism ; Free Radicals ; Gene Expression Regulation ; Glutathione/METABOLISM ; Glutathione Reductase/METABOLISM ; Glutathione Transferases/ METABOLISM ; Human ; Longevity ; Oxidation-Reduction ; Oxygen Consumption ; Review ; Superoxide Dismutase/METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Temperature ; Thermodynamics SO - Dermatol Clin 1986 Jul;4(3):347-58 25 UI - 86241232 AU - Kirsch JR ; Dean JM ; Rogers MC TI - Current concepts in brain resuscitation. AB - In spite of the tremendous amount of effort and money put forth to reduce morbidity and mortality associated with global cerebral ischemia, the outlook for patients suffering an ischemic insult remains dismal. The lack of a sufficient substrate supply during the period of ischemia as well as the production of toxic metabolites in response to ischemia have been incriminated as key factors causing brain damage. As discussed in this article, modes of therapy have included efforts to minimize the duration of ischemia (eg, effective cardiopulmonary resuscitation, hemodilution, heparinization, calcium antagonists) and decrease the production of toxic metabolites (eg, barbiturates, calcium antagonists). Although the barbiturates have also been proposed to decrease the metabolic needs during ischemia, they have no therapeutic value for global cerebral ischemia. The initial evaluation of the calcium antagonists has been more promising. MH - Animal ; Barbiturates/THERAPEUTIC USE ; Brain/METABOLISM/ PATHOLOGY ; Calcium/ANALYSIS ; Calcium Channel Blockers/ THERAPEUTIC USE ; Cerebral Ischemia/ETIOLOGY/*THERAPY ; Cerebrovascular Circulation ; Disease Models, Animal ; Etomidate/ THERAPEUTIC USE ; Fatty Acids/ANALYSIS ; Free Radicals ; Heart Arrest/COMPLICATIONS ; Hemodilution ; Heparin/THERAPEUTIC USE ; Human ; Lactates/ANALYSIS ; Mitochondria/ANALYSIS ; *Resuscitation ; Review SO - Arch Intern Med 1986 Jul;146(7):1413-9 26 UI - 86241098 AU - Pieri C ; Giuli C ; Bertoni-Freddari C ; Bernardini A TI - Vitamin E deficiency alters the in vivo Rb+ discrimination of rat brain cortical cells. AB - The in vivo Rb+ uptake and release of rat brain cortical cells of 11-months-old rats fed with a vitamin E deficient diet was investigated. The animals were treated with a daily dose of 30 mg RbCl/100 g body weight for 14 days. After discontinuation of the RbCl treatment the animals were killed at intervals of 2, 4, 9 and 15 days, respectively. The intracellular Rb+ and K+ contents were analyzed by energy dispersive X-ray microanalysis, whereas concentrations of these two ions were determined by atomic absorbtion spectrophotometry in the serum and cerebrospinal fluid. Vitamin E deficient rats accumulate more Rb+ than age-matched normally fed animals at any time taken into account. Rb+-discrimination ratios calculated on the basis of Rb+ and K+ contents of both, cortical cell cytoplasm and cerebrospinal fluid, are higher in vitamin E deficient rats than in the controls (+20%), which supports the view that the enhanced membrane lipid peroxidation induced by vitamin E deficiency impairs the passive membrane permeability for Rb+ (and K+). MH - Aging ; Animal ; Cell Membrane Permeability ; Cerebral Cortex/ *METABOLISM ; Female ; Free Radicals ; Lipid Peroxides/ BIOSYNTHESIS ; Membrane Lipids/METABOLISM ; Potassium/METABOLISM ; Rats ; Rats, Inbred Strains ; Rubidium/*METABOLISM ; Support, Non-U.S. Gov't ; Vitamin E Deficiency/*METABOLISM SO - Arch Gerontol Geriatr 1986 Apr;5(1):21-31 27 UI - 86239626 AU - Parks DA ; Granger DN TI - Contributions of ischemia and reperfusion to mucosal lesion formation. AB - Two theories have been proposed to account for the mucosal injury associated with intestinal ischemia, hypoxia-countercurrent exchange, and oxygen free radicals. The countercurrent mechanism suggests that mucosal injury should occur predominately during the ischemic period, whereas the oxygen free radical hypothesis predicts that the majority of mucosal injury results from reperfusion of ischemic tissue. Histological specimens obtained during the ischemic period and following reperfusion allowed a systematic evaluation of the time course of development of mucosal lesions in a regional ischemia model. Reperfusion after 3 h of regional hypotension reduced mean mucosal thickness from 1,022.2 +/- 6.3 to 503.6 +/- 10.0 microns. The decrease in mucosal thickness was largely due to a reduction in villus height, inasmuch as the reduction in crypt depth was statistically insignificant. A significantly smaller change in mucosal thickness was observed when the bowel was subjected to 3 h ischemia without reperfusion. The mucosal injury produced by 3 h ischemia and 1 h reperfusion was more severe than that produced by 4 h ischemia without reperfusion. The results of this study suggest that most of the tissue damage produced by the widely employed regional hypotension model occurs at the time of reperfusion. MH - Animal ; Cats ; Epithelium/PATHOLOGY ; Free Radicals ; Ileum/ *BLOOD SUPPLY/PATHOLOGY ; Intestinal Mucosa/*BLOOD SUPPLY/ PATHOLOGY ; Ischemia/*PHYSIOPATHOLOGY ; Necrosis ; Oxygen/ METABOLISM ; Support, U.S. Gov't, P.H.S. ; Time Factors SO - Am J Physiol 1986 Jun;250(6 Pt 1):G749-53 28 UI - 86225520 AU - Gerberick GF ; Jaffe HA ; Willoughby JB ; Willoughby WF TI - Relationships between pulmonary inflammation, plasma transudation, and oxygen metabolite secretion by alveolar macrophages. AB - We have previously shown that alveolar macrophages from normal rabbit lungs do not synthesize reactive oxygen intermediates unless first conditioned by culture in vitro in the presence of serum for 24 to 48 hr. This conditioning process is mediated by a serum constituent that partitions on gel exclusion columns with an apparent m.w. of 30,000 to 50,000 daltons. Alveolar macrophage conditioning in vitro requires protein synthesis, is associated with the generation of membrane NADPH oxidase activity, and is reversible. We have predicted therefore that during the course of pulmonary inflammation, as observed 3 wk after i.v. injection of M. butyricum in oil, alveolar macrophages might similarly become conditioned in vivo through exposure to plasma protein transudates reaching the alveolus. In support of this hypothesis we show that after experimental production of granulomatous pulmonary inflammation in rabbits, alveolar macrophages showed an augmented capacity to secrete superoxide anion when stimulated with phorbol ester, and this enhancement increases exponentially with increased plasma transudation. This augmented enhancement was reversible, and decreased after culture in vitro in the absence of serum. Mature alveolar macrophages were responsible for this enhanced superoxide anion production rather than freshly emigrated monocytes. Moreover, superoxide anion production in this model of pulmonary inflammation appears to be an "all-or-none: phenomenon, with superoxide anion production associated with a subpopulation of optimally conditioned alveolar macrophages, whereas the remaining unconditioned alveolar macrophages produce little or none. We feel that these two classes of alveolar macrophages may be derived from inflamed and noninflamed regions of the lung, respectively, thereby reflecting the discontinuous nature of the inflammatory lesions themselves. Thus we propose that measurements of reactive oxygen intermediate production by lavaged alveolar macrophages may provide a semi-quantitative measure of chronic pulmonary inflammation. MH - Animal ; *Capillary Permeability ; Cell Adhesion ; Free Radicals ; Inflammation/BLOOD/*PATHOLOGY/PHYSIOPATHOLOGY ; Lung/METABOLISM/ *PATHOLOGY ; Macrophages/*METABOLISM ; Male ; Monocytes/ PHYSIOLOGY ; Oxygen/*METABOLISM ; Pulmonary Alveoli/CYTOLOGY ; Rabbits ; Rats ; Rats, Inbred LEW ; Rats, Inbred Strains ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - J Immunol 1986 Jul 1;137(1):114-21 29 UI - 86224906 AU - Ward PA ; Johnson KJ ; Till GO TI - Complement and experimental respiratory failure. AB - Activation of the complement system within the lung can lead to acute pulmonary damage and dysfunction. Based on a variety of experimental models it is now apparent that lung injury is related to complement-induced generation of oxygen derived free radicals from neutrophils and from macrophages. In addition to the oxygen radicals, it is also possible that the conversion of hydrogen peroxide by myeloperoxidase to hypochlorous acid also contributes to the injury. Exposure of the pulmonary microvasculature to oxygen radicals generated from complement-activated neutrophils causes focal damage and necrosis of endothelial cells. IgG immune complex-induced injury of lung is also complement and neutrophil dependent and oxygen radical mediated. In contrast, lung injury produced by IgA immune complexes is neutrophil independent, complement dependent and oxygen radical mediated. There is now increasing evidence that oxygen radicals are not only directly tissue-toxic but also able to potentiate the activity of leukocytic proteases. In all of these models the lung can be protected from injury by pretreatment of the animals with either scavengers of hydroxyl radical or with agents that prevent its formation (e.g. catalase, iron chelators). Data from these models may have direct clinical relevance to conditions such as adult respiratory distress syndrome where lung injury is probably oxygen radical mediated. MH - Animal ; Catalase/THERAPEUTIC USE ; *Complement Activation ; Free Radicals ; Human ; IgA/METABOLISM ; IgG/METABOLISM ; Iron Chelates/THERAPEUTIC USE ; Lung/DRUG EFFECTS/METABOLISM/ PHYSIOPATHOLOGY ; Macrophages/METABOLISM ; Models, Biological ; Neutrophils/METABOLISM ; Rabbits ; Respiratory Insufficiency/DRUG THERAPY/*IMMUNOLOGY/PHYSIOPATHOLOGY ; Sheep ; Superoxide/ METABOLISM/TOXICITY SO - Intensive Care Med 1986;12(1):17-21 30 UI - 86218943 AU - Das DK ; Engelman RM ; Otani H ; Rousou JA ; Breyer RH ; Lemeshow S TI - Effect of superoxide dismutase and catalase on myocardial energy metabolism during ischemia and reperfusion. AB - Survival of cardiac patients undergoing heart surgery depends critically upon the recovery of myocardial energy metabolism during reperfusion of ischemic myocardium. The present study compares various parameters of myocardial energy metabolism using an isolated in situ pig heart. The left anterior descending (LAD) coronary artery was occluded for 60 min, followed by 60 min of global hypothermic cardioplegic arrest and 60 min of reperfusion. Free radical scavengers [superoxide dismutase SOD and catalase] were used to protect the ischemic heart from reperfusion injury. In both control and SOD plus catalase-treated groups, ATP, creatine phosphate (CP), ATP/ADP ratio, energy charge and phosphorylation potential dropped significantly during ischemic insult. After reperfusion, CP, ATP/ADP ratio and phosphorylation potential improved significantly, but they were restored to control level only in treated animals. In either case, free energy of ATP hydrolysis (delta G) lowered only by 5% during ischemia, but recovered promptly upon reperfusion. SOD and catalase also improved coronary blood flow and reduced creatine kinase release compared to those of untreated animals, suggesting improved myocardial recovery upon reperfusion. Our results suggest that SOD and catalase significantly improve the myocardial recovery during reperfusion by enhancing rephosphorylation steps, and the value of delta G is more critical compared to those of ATP and CP for myocardial recovery. MH - Adenosine Triphosphate/METABOLISM ; Animal ; Catalase/ *PHARMACODYNAMICS ; Comparative Study ; Coronary Circulation/DRUG EFFECTS ; Coronary Disease/*DRUG THERAPY/ENZYMOLOGY/METABOLISM/ PHYSIOPATHOLOGY ; Creatine Kinase/BLOOD/METABOLISM ; Energy Metabolism/*DRUG EFFECTS ; Female ; Free Radicals ; Heart/DRUG EFFECTS ; In Vitro ; Male ; Myocardium/ENZYMOLOGY/*METABOLISM ; Perfusion ; Superoxide Dismutase/*PHARMACODYNAMICS ; Swine SO - Clin Physiol Biochem 1986;4(3):187-98 31 UI - 86214274 AU - Coles JC ; Ahmed SN ; Mehta HU ; Kaufmann JC TI - Role of free radical scavenger in protection of spinal cord during ischemia. AB - Previous work in our laboratory established an experimental model for the production of paraplegia in the anesthetized normothermic adult mongrel dog. The current study involves 24 animals divided into two equal groups: Group 1 served as control, and Group 2 received treatment with scavenger agent. Vascular occlusive clamps were placed on the thoracic aorta proximal to the left subclavian artery, on the left subclavian artery at its origin, and on the distal thoracic aorta at the diaphragm for 30 minutes. In Group 1,200 ml of normal saline solution (37 degrees C) was perfused into the occluded aortic segment at the rate of 0.33 ml per kilogram of body weight per minute. In Group 2, 90% dimethyl sulfoxide (DMSO) in a dose of 0.1 gm/kg in normal saline solution (37 degrees C) for a total volume of 200 ml, was likewise injected into the occluded aortic segment at the same infusion rate. Animals were observed for evidence of paresis in the postoperative period. Microscopic analysis revealed evidence of ischemic myelopathy in the control group but none in the treated group. Under the conditions of this experiment, we conclude that the scavenger agent DMSO has a highly protective effect on the spinal cord during ischemic insult. MH - Animal ; Aorta, Thoracic/*SURGERY ; Dimethyl Sulfoxide/ ADMINISTRATION & DOSAGE/*PHARMACODYNAMICS ; Dogs ; Free Radicals ; Ischemia/*PATHOLOGY ; Microscopy, Electron, Scanning ; Nuclear Magnetic Resonance ; Paresis/PREVENTION & CONTROL ; Spinal Cord/ *BLOOD SUPPLY/DRUG EFFECTS/PATHOLOGY ; Subclavian Artery SO - Ann Thorac Surg 1986 May;41(5):551-6 32 UI - 86211665 AU - Del Soldato P ; Foschi D ; Benoni G ; Scarpignato C TI - Oxygen free radicals interact with indomethacin to cause gastrointestinal injury. AB - In the present study it was shown that, unlikely MK447, a known oxygen free radical compound, PGE2 is much less effective against indomethacin-induced G.I. ulcers than against ethanol damage. It seems likely that factors other than PG deficiency (such as oxygen free radicals), could be involved in the pathogenesis of NSAID-induced G.I. damage. Some compounds that can capture free radicals (aminopyrine, thiourea and its derivative, MK 447) or that inhibit the lipoxygenase pathway (MK 447, salicylazosulfapyridine, BW 755, benoxaprofen) are able to abolish indomethacin-induced G.I. damage. After irradiation with hydroxyl free radicals, indomethacin reacts with them to cause marked G.I. injury, even at a submaximal dose, one poorly ulcerogenic by itself. The above findings suggest that oxygen free radicals are one of the causal factors in the formation of NSAID-induced G.I. side effects. Some of the data in this paper were presented at Fermo, August 31, 1984 (Advanced course on 'oxygen and sulfur radicals in chemistry and medicine') and at the 9th Iuphar International Congress of Pharmacology in London, July 30, 1984. MH - Alcohol, Ethyl/TOXICITY ; Animal ; Anti-Inflammatory Agents/ *PHARMACODYNAMICS ; Butylated Hydroxytoluene/*ANALOGS & DERIVATIVES/PHARMACODYNAMICS ; Female ; Free Radicals ; Gastrointestinal System/DRUG EFFECTS/*PATHOLOGY ; Indomethacin/ *TOXICITY ; Oxygen/*TOXICITY ; Peptic Ulcer/*CHEMICALLY INDUCED ; Prostaglandins E/*PHARMACODYNAMICS ; Rats ; Stomach Ulcer/ *CHEMICALLY INDUCED SO - Agents Actions 1986 Mar;17(5-6):484-8 33 UI - 86203000 AU - Garfinkel D TI - Is aging inevitable? The intracellular zinc deficiency hypothesis of aging. AB - A review of the literature suggests that an intracellular zinc deficiency may be the primary cause of the aging process. Zinc-metalloenzymes play an important role in many aspects of cellular metabolism including DNA replication, repair and transcription. The main enzymes affected by zinc deficiency may be specific for each cell type. Depending on which zinc enzymes are "overvulnerable:, zinc deficiency may result in accumulation of useless (or toxic) materials, malproduction of essential proteins, a neoplastic change or cell death, thus explaining the variability in aging patterns in different cell types. There is no simple and reliable index of zinc status in humans and a therapeutic trial may be needed to establish zinc deficiency. Finding a zinc-compound which can enter the cell and avoid the development of intracellular zinc deficiency may retard the aging process and postpone age-related diseases. MH - *Aging ; Animal ; Cardiovascular Diseases/ETIOLOGY ; Cells/ *PHYSIOLOGY ; Cells, Cultured ; Dementia, Senile/ETIOLOGY ; Diabetes Mellitus/ETIOLOGY ; DNA/PHYSIOLOGY ; Environment ; Enzymes/PHYSIOLOGY ; Female ; Forecasting ; Free Radicals ; Genetics ; Human ; Immune System/PHYSIOPATHOLOGY ; Intestinal Absorption ; Metalloproteins/PHYSIOLOGY ; *Models, Biological ; Neoplasms/ETIOLOGY ; Pregnancy ; Pregnancy Complications ; Review ; Zinc/ADMINISTRATION & DOSAGE/*DEFICIENCY/PHYSIOLOGY/THERAPEUTIC USE SO - Med Hypotheses 1986 Feb;19(2):117-37 34 UI - 86201586 AU - Johnson KJ ; Ward PA ; Kunkel RG ; Wilson BS TI - Mediation of IgA induced lung injury in the rat. Role of macrophages and reactive oxygen products. AB - Acute lung injury in the rat has been induced by the instillation of affinity-purified mouse monoclonal IgA antibody with specific reactivity to the hapten dinitrophenol coupled to albumin. As previously reported, this model of lung injury requires an intact complement system but is independent of neutrophils. In contrast to macrophages obtained by bronchoalveolar lavage from rats receiving IgA into the airways in the absence of intravenously injected antigen, macrophages obtained from the lungs of rats developing IgA immune complex-induced lung injury were significantly increased in number, showed greater spontaneous generation of O2.-, and demonstrated significantly enhanced O2.- responses in the presence of an added stimulus, phorbol ester. Inhibition studies in vivo suggested that the IgA-induced lung injury is mediated by oxygen radicals generated from lung macrophages. Pretreatment of animals with superoxide dismutase, catalase, the iron chelator, deferoxamine, or the hydroxyl radical scavenger, dimethyl sulfoxide, suppressed the development of lung injury. Morphologically the lungs of protected animals showed increased numbers of mononuclear cells within the alveolar compartment but little evidence of alveolar or capillary injury, in contrast to unprotected animals in which there was evidence of severe injury, both to microvascular interstitial endothelial cells as well as to alveolar lining epithelial cells. These studies suggest that acute lung injury in the rat induced by IgA immune complexes is mediated by oxygen radical formation and that the macrophage may be the principle effector cell, as compared to IgG immune complex induced lung injury, which is also oxygen radical mediated but in which the neutrophil is the effector cell. MH - Animal ; Catalase/PHARMACODYNAMICS ; Deferoxamine/ PHARMACODYNAMICS ; Dimethyl Sulfoxide/PHARMACODYNAMICS ; Dinitrophenols/IMMUNOLOGY ; Free Radicals ; Haptens/IMMUNOLOGY ; IgA/*IMMUNOLOGY ; Immune Complex Disease/*IMMUNOLOGY/METABOLISM ; Lung/PATHOLOGY ; Lung Diseases/*IMMUNOLOGY/METABOLISM/PATHOLOGY ; Macrophages/*IMMUNOLOGY/METABOLISM ; Pulmonary Alveoli/PATHOLOGY ; Rats ; Serum Albumin, Bovine/IMMUNOLOGY ; Superoxide/ *METABOLISM ; Superoxide Dismutase/PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Lab Invest 1986 May;54(5):499-506 35 UI - 86201289 AU - Diamond JR ; Bonventre JV ; Karnovsky MJ TI - A role for oxygen free radicals in aminonucleoside nephrosis. AB - The cellular processes responsible for the proteinuria induced by the aminonucleoside of puromycin (PA) remain inadequately defined. Hypoxanthine is both a metabolic breakdown product of PA as well as a substrate for xanthine oxidase, which catalyzes its enzymatic conversion to xanthine and uric acid, yielding the superoxide anion in the process. We examined whether oxygen free radical production contributes to the development of proteinuria in this model. Seven groups of male Sprague-Dawley rats were studied. Proteinuria was quantitated and histology examined 7 days after rats were treated with PA intravenously over 5 min. PA-treated animals received either saline, dimethyl sulfoxide, superoxide dismutase, or catalase over 30 min prior to and 30 min following PA administration. Another group received allopurinol over 4 hr prior to PA. The superoxide dismutase and allopurinol treatment groups had a significant suppression of urinary protein excretion compared to the PA control group. There were also less severe glomerular morphologic changes in the superoxide dismutase group vs. the PA controls, which demonstrated a pathologic pattern that included epithelial cell blebbing, segmental mesangial cell proliferation and matrix expansion, loss of glomerular capillary lumina, and occasional adhesions between the glomerular tuft and Bowman's capsule. The allopurinol group exhibited normal glomerular morphology on light microscopy, with the exception of occasional epithelial cell blebs. All groups showed spreading of the epithelial cell cytoplasm along the glomerular basement membrane with loss of foot processes, focal areas of lifting of the epithelial cell from the glomerular basement membrane, cytoplasmic vacuolization, and protein reabsorption droplets; however, allopurinol-treated animals demonstrated these changes to a lesser extent.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Allopurinol/PHARMACODYNAMICS ; Animal ; Catalase/PHARMACODYNAMICS ; Depression, Chemical ; Dimethyl Sulfoxide/PHARMACODYNAMICS ; Disease Models, Animal ; Free Radicals ; Male ; Nephrosis/ *CHEMICALLY INDUCED/PATHOLOGY ; Oxygen/*PHYSIOLOGY ; Proteinuria/ ETIOLOGY ; *Puromycin/ANALOGS & DERIVATIVES ; *Puromycin Aminonucleoside ; Rats ; Rats, Inbred Strains ; Superoxide Dismutase/PHARMACODYNAMICS ; Support, U.S. Gov't, P.H.S. SO - Kidney Int 1986 Feb;29(2):478-83 36 UI - 86198922 AU - Kontos HA ; Wei EP TI - Superoxide production in experimental brain injury. AB - The appearance of superoxide anion radicals in cerebral extracellular space during and after experimental fluid-percussion brain injury was investigated in anesthetized cats equipped with cranial windows. Superoxide was detected by demonstrating the presence of superoxide dismutase (SOD)-inhibitable reduction of nitroblue tetrazolium (NBT). The SOD-inhibitable rate of reduction of NBT was 3.52 +/- 0.72 nM/min/sq cm during brain injury and 4.11 +/- 0.74 nM/min/sq cm 1 hour after injury. No significant superoxide production was detected in control animals. The sustained arteriolar dilation and reduced responsiveness to the vasoconstrictor effects of arterial hypocapnia observed 30 minutes after brain injury were eliminated by after-treatment with topical SOD (60 U/ml) and catalase (40 U/ml). The results show that experimental brain injury causes the generation and appearance in extracellular fluid space of superoxide. Superoxide production continues for at least 1 hour following injury. The sustained dilation and abnormal responsiveness of cerebral arterioles after injury are due to the continued generation of superoxide and other radicals derived from it. These functional changes can be reversed by after-treatment with appropriate scavenging agents. MH - Animal ; Arachidonic Acids/METABOLISM ; Brain Injuries/ *METABOLISM/PHYSIOPATHOLOGY ; Cats ; Free Radicals/METABOLISM ; Oxygen/*METABOLISM ; Prostaglandin Synthase/METABOLISM ; Superoxide/*METABOLISM ; Superoxide Dismutase/METABOLISM ; Support, U.S. Gov't, P.H.S. SO - J Neurosurg 1986 May;64(5):803-7 37 UI - 86198352 AU - Asayama K ; Kooy NW ; Burr IM TI - Effect of vitamin E deficiency and selenium deficiency on insulin secretory reserve and free radical scavenging systems in islets: decrease of islet manganosuperoxide dismutase. AB - There is increasing evidence that islet beta cells may be susceptible to redox insult, and that this susceptibility may contribute to the pathogenesis of experimental models of diabetes mellitus. We investigated the effect of vitamin E deficiency, selenium deficiency, and combined deficiency on islet function and free radical scavenging systems. The tissue levels of glutathione peroxidase, catalase, and immunoreactive superoxide dismutases were measured in four groups of rats (i.e., controls and those with vitamin E, selenium, and combined deficiency). Glucose tolerance tests were performed for each animal before sacrifice. Superoxide dismutase concentrations in liver, heart, and skeletal muscle were within 20% of the control levels in all groups. However, the manganosuperoxide dismutase concentrations in islets were significantly lower than control levels in response to vitamin E, selenium, and combined deficiency. Combined deficiency appeared to have an additive effect. In contrast, cuprozinc superoxide dismutase concentration in islets was higher in the deficient groups than in controls. Insulin secretory reserve was decreased in each of the three deficient groups. This decrease was reflected as glucose intolerance only in the group with combined deficiency. Glutathione peroxidase activity was markedly decreased in selenium-deficient animals in all tissues studied. Catalase activity did not change significantly among groups in any tissue studied. Islets had the lowest glutathione peroxidase and cuprozinc and total superoxide dismutase levels among tissues studied. MH - Animal ; Blood Glucose/ANALYSIS ; Catalase/METABOLISM ; Free Radicals ; Glucose Tolerance Test ; Glutathione Peroxidase/ METABOLISM ; Insulin/*SECRETION ; Islands of Langerhans/ ENZYMOLOGY/METABOLISM/*SECRETION ; Liver/ENZYMOLOGY ; Male ; Manganese/*METABOLISM ; Muscles/ENZYMOLOGY ; Myocardium/ ENZYMOLOGY ; Rats ; Rats, Inbred Strains ; Selenium/*DEFICIENCY ; Superoxide Dismutase/*METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Vitamin E Deficiency/COMPLICATIONS/ METABOLISM/*PHYSIOPATHOLOGY SO - J Lab Clin Med 1986 May;107(5):459-64 38 UI - 86196856 AU - Campbell CA ; Przyklenk K ; Kloner RA TI - Infarct size reduction: a review of the clinical trials. AB - The most important finding to emerge from this review of experimental and clinical studies is that the earlier therapy is begun after the onset of symptoms of acute MI, the greater the potential for reduction of infarct size and possibly mortality. It is difficult to define a precise time after which therapy would not have an effect, since the clinical trials for each drug group vary significantly in respect to time of therapy initiation. In experimental studies, major salvage of ischemic myocardium occurs when the drug is given within two hours of coronary artery occlusion. If drug therapy is begun four to six hours postocclusion, then only minor or no reductions in infarct size will occur. The ability of any drug or intervention to reduce infarct size in humans would be optimized if therapy were begun less than four hours of onset of symptoms. With the realization of the wavefront phenomenon and the potential salvage of myocardium at risk with reperfusion, the introduction of reperfusion in the clinical setting with thrombolytic agents or other procedures becomes highly desirable. Clot-selective thrombolytic agents, such as tissue plasminogen activator, diminish the adverse effects and high costs of intracoronary thrombolytic therapy or PTCA. Consequently, it is probable that the initial procedure of choice would be the use of clot-selective thrombolytic therapy. Thrombolytic therapy only lyses thrombi and does not affect the underlying causes of the coronary artery occlusion. Therefore, therapy to reduce the chances of reinfarction and death must also be initiated. Percutaneous transluminal coronary angioplasty, in selected patients, should reduce the reocclusion rate. Beta-adrenoceptor blocking agents appear to be an excellent therapy for reducing mortality when administered chronically; these agents reduce myocardial oxygen consumption and reverse the imbalance between oxygen supply and oxygen demand caused by activation of the sympathetic nervous system and actions of catecholamines. Since thrombus formation has occurred at least once in patients who survive an MI, it is probable that the conditions for thrombus formation still exist. Therefore, institution of antiplatelet aggregating drugs, such as aspirin, would seem to be an appropriate prophylactic regimen. Beta blockers and possibly nitroglycerin have desirable effects when thrombolysis is unavailable. The efficacy of calcium-channel blocking agents on reduction of infarct size appears to be limited, although in the setting of stable and unstable angina postinfarction, these agents can play an important role.(ABSTRACT TRUNCATED AT 400 WORDS) MH - Adrenergic Beta Receptor Blockaders/THERAPEUTIC USE ; Animal ; Anti-Inflammatory Agents/THERAPEUTIC USE ; Calcium Channel Blockers/THERAPEUTIC USE ; Clinical Trials ; Coronary Disease/ PHYSIOPATHOLOGY ; Creatine Kinase Isoenzymes/ANALYSIS ; Dogs ; Electrocardiography ; Fibrinolytic Agents/THERAPEUTIC USE ; Free Radicals ; Glyceryl Trinitrate/THERAPEUTIC USE ; Human ; Hyaluronidase/THERAPEUTIC USE ; Myocardial Infarction/*DRUG THERAPY/PATHOLOGY/PHYSIOPATHOLOGY ; Necrosis/PATHOLOGY ; Oxygen/ METABOLISM ; Perfusion ; Support, Non-U.S. Gov't SO - J Clin Pharmacol 1986 May-Jun;26(5):317-29 39 UI - 86196219 AU - Zaizen Y ; Nakagawara A ; Ikeda K TI - Patterns of destruction of mouse neuroblastoma cells by extracellular hydrogen peroxide formed by 6-hydroxydopamine and ascorbate. AB - The patterns of the cytolytic effects of 6-hydroxydopamine (6-OHDA), with/without ascorbate, on C-1300 and three other cloned mouse neuroblastoma cell lines (N1E-115, NS-20, N-18) were studied in vitro. The sensitivity to 6-OHDA differed and the three cloned cell lines were more sensitive than the wild type C-1300 cell line. Ascorbate synergistically potentiated the cytolytic effect of 6-OHDA to all four cell lines. The 6-OHDA cytotoxicity was eliminated by the addition of exogenous catalase but not by addition of other oxygen free radical scavengers, thereby suggesting that the hydrogen peroxide formed might influence the cells, extracellularly. In addition, the critical time for tumor cell lysis was the first 60 min of the reaction. The cytotoxicity induced by the unmasked cyclophosphamide, 4-hydroperoxycyclophosphamide, was synergistically enhanced in the presence of a nontoxic concentration of 6-OHDA and ascorbate. These data suggest that reactive oxygen intermediates may prove to be a good tool for destroying neuroblastoma cells. MH - Animal ; Antineoplastic Agents, Combined/THERAPEUTIC USE ; Ascorbic Acid/METABOLISM/TOXICITY/*THERAPEUTIC USE ; Cell Line ; Cisplatin/ADMINISTRATION & DOSAGE ; Clone Cells/DRUG EFFECTS/ METABOLISM ; Comparative Study ; Doxorubicin/ADMINISTRATION & DOSAGE ; Drug Screening ; Drug Synergism ; Female ; Free Radicals ; Hydrogen Peroxide/METABOLISM/*THERAPEUTIC USE ; Hydroxydopamines/METABOLISM/TOXICITY/*THERAPEUTIC USE ; Mice ; Mice, Inbred A ; Neuroblastoma/*DRUG THERAPY/METABOLISM/PATHOLOGY ; Spleen/DRUG EFFECTS ; Time Factors ; Vincristine/ADMINISTRATION & DOSAGE SO - J Cancer Res Clin Oncol 1986;111(2):93-7 40 UI - 86190438 AU - Mitsos SE ; Askew TE ; Fantone JC ; Kunkel SL ; Abrams GD ; Schork A ; Lucchesi BR TI - Protective effects of N-2-mercaptopropionyl glycine against myocardial reperfusion injury after neutrophil depletion in the dog: evidence for the role of intracellular-derived free radicals. AB - Reperfusion of the previously ischemic myocardium is associated with the production of oxygen free radicals and their metabolites, which contribute to the ultimate extent of irreversible myocardial injury. The relative importance of polymorphonuclear leukocytes vs intracellular-derived oxygen metabolites has remained uncertain. We evaluated the effectiveness of a free-radical scavenger, N-2-mercaptopropionyl glycine (MPG), in limiting infarct size after ischemia/reperfusion in dogs that were depleted of neutrophils with specific antisera. Twenty-four urethane-anesthetized open-chest dogs were subjected to 90 min of ischemia by occlusion of the left circumflex coronary artery followed by 6 hr of reperfusion. Dogs were randomly assigned to receive nonimmune serum, neutrophil antiserum, or neutrophil antiserum plus MPG (20 mg/kg intra-atrially 15 min before reperfusion was initiated and for 45 min after reperfusion). Infarct size, as a percent of the area at risk, was reduced by 33% in the neutrophil antiserum group as compared with the nonimmune group (30.7 +/- 2.7% vs 45.6 +/- 3.7%, p less than .01). The combined administration of neutrophil antiserum plus MPG reduced the size of infarction by 63% of the area at risk compared with that in the nonimmune group (17.0 +/- 2.7% vs 45.6 +/- 3.7%, p less than .01). The reduction in infarct size with neutrophil antiserum plus MPG was significantly greater than that with the neutrophil antiserum alone (p less than .01). The areas at risk did not differ among the groups. Myocardial protection could not be explained on the basis of hemodynamic differences. The observation that MPG enhances the protective effects of neutrophil depletion suggests that both extramyocardial- and intramyocardial-derived oxygen free radicals contribute significantly to reperfusion-induced myocardial injury. MH - Amino Acids, Sulfur/*THERAPEUTIC USE ; Animal ; Cell Movement/ DRUG EFFECTS ; Dogs ; Free Radicals ; Hemodynamics/DRUG EFFECTS ; Male ; Mercaptopropionylglycine/PHARMACODYNAMICS/*THERAPEUTIC USE ; Myocardial Infarction/*DRUG THERAPY/METABOLISM/PATHOLOGY ; Myocardium/METABOLISM/PATHOLOGY ; Neutropenia/ETIOLOGY/METABOLISM/ PATHOLOGY ; Neutrophils/*METABOLISM/PHYSIOLOGY ; Oxygen Consumption/*DRUG EFFECTS ; *Perfusion ; Random Allocation ; Time Factors SO - Circulation 1986 May;73(5):1077-86 41 UI - 86189629 AU - O'Connell JF ; Klein-Szanto AJ ; DiGiovanni DM ; Fries JW ; Slaga TJ TI - Enhanced malignant progression of mouse skin tumors by the free-radical generator benzoyl peroxide. AB - Chemical carcinogenesis in mouse skin can be divided into the processes of initiation, promotion, and progression. The free-radical generator benzoyl peroxide is moderately active during the promotion stage. Repetitive treatment of mouse benign skin tumors (papillomas) with benzoyl peroxide (20 mg, twice weekly) increased the number of cumulative carcinomas per group by 325% and the number of keratoacanthomas by 44% compared to tumor-bearing Sencar mice treated with the promoter 12-O-tetradecanoylphorbol-13-acetate. The lack of increase in the number of cumulative papillomas per group due to benzoyl peroxide treatment suggests that benzoyl peroxide enhanced the progression of preexisting papillomas. The ability of benzoyl peroxide to enhance the progression of benign tumors to cancer should be considered when determining the human risk from exposure to this widely used chemical agent; in addition, biological assays specifically testing malignant progression may be essential and beneficial for determining an agent's carcinogenic risk. MH - Animal ; Benzoyl Peroxide/*TOXICITY ; Cocarcinogenesis ; Female ; Free Radicals ; Keratoacanthoma/CHEMICALLY INDUCED ; Mice ; Mice, Inbred Strains ; Papilloma/CHEMICALLY INDUCED ; Peroxides/ *TOXICITY ; Skin Neoplasms/*CHEMICALLY INDUCED/PATHOLOGY ; Support, U.S. Gov't, P.H.S. ; Tetradecanoylphorbol Acetate ; 9,10-Dimethyl-1,2-Benzanthracene SO - Cancer Res 1986 Jun;46(6):2863-5 42 UI - 86189605 AU - Carr BI ; Langley D ; Dias CB ; Hammond WG ; Benfield JR TI - Differential interaction of normal and preneoplastic hamster bronchi with adriamycin. AB - Advanced bronchogenic carcinoma in humans is notoriously resistant to the cytocidal actions of cancer chemotherapy. The experiments reported here were undertaken as a first step in examining the mechanisms of resistance of carcinogen-altered bronchus to the actions of the commonly used cancerocidal agent Adriamycin. Syrian Golden hamsters were treated with an endobronchial carcinogen in order to produce bronchial neoplasms or with no carcinogen as controls. Hamsters were then given i.v. Adriamycin, and the amounts and metabolism of bronchial Adriamycin were determined. Peak uptake values were found 5 min after Adriamycin administration, and the amounts of Adriamycin in normal and carcinogen-altered bronchi were found to be similar. Whereas no metabolism of Adriamycin was observed in normal bronchi, 40-60% of total Adriamycin fluorescence was found to be due to Adriamycinol and Adriamycin aglycones in bronchi with premalignant changes. In separate experiments, the susceptibility of normal and carcinogen-altered bronchial extracts to drug-induced lipid peroxidation was measured in vitro. A 50% decrease was found in the ability of carcinogen-altered bronchi to act as a substrate for lipid peroxidation mediated by Adriamycin and an approximately 30% decrease for lipid peroxidation induced by t-butyl-hydroperoxide. These results demonstrate two different mechanisms by which bronchogenic carcinomas might become resistant to the chemotherapeutic actions of Adriamycin. These are by the carcinogen induction of metabolism of Adriamycin to less toxic products and by resistance of the bronchi to free radical damage. MH - Animal ; Bronchi/*DRUG EFFECTS/METABOLISM/PATHOLOGY ; Bronchial Neoplasms/DRUG THERAPY/*METABOLISM ; Doxorubicin/METABOLISM/ *PHARMACODYNAMICS ; Free Radicals ; Hamsters ; Lipid Peroxides/ METABOLISM ; Mesocricetus ; Precancerous Conditions/DRUG THERAPY/ *METABOLISM ; Support, U.S. Gov't, P.H.S. SO - Cancer Res 1986 Jun;46(6):2730-4 43 UI - 86189548 AU - Yamashina K ; Miller BE ; Heppner GH TI - Macrophage-mediated induction of drug-resistant variants in a mouse mammary tumor cell line. AB - The ability of macrophages to induce drug-resistant variants was studied in an in vitro macrophage-tumor cell coculture system utilizing the hypoxanthine-guanine phosphoribosyl transferase locus as measured by resistance to 6-thioguanine. Tumor cells of mouse mammary tumor line 66 were sensitive to macrophage induction of thioguanine resistance as shown by an increase in the frequency of thioguanine-resistant variants which arose following macrophage coculture to levels at least 5- to 10-fold above the spontaneous frequency. Detection of increased numbers of variants depended upon the macrophage:tumor cell ratio, with 50:1 or greater being necessary. The activity of the macrophages was dependent upon their activation stage. The induction of drug-resistant variants could be inhibited by oxygen radical scavengers. The basis for the emergence of thioguanine-resistant cells appeared to be induction of new variants rather than selection of preexisting resistant cells from the parental population, since thioguanine-sensitive and -resistant cells were equally sensitive to macrophage-mediated toxicity. In six of the six macrophage-induced variants tested, resistance was associated with loss of hypoxanthine-guanine phosphoribosyl transferase activity. The reverse variation frequency at the hypoxanthine-guanine phosphoribosyl transferase locus in five macrophage-induced variants was low and similar to that of a stable ethyl methanesulfonate-induced, thioguanine-resistant line. Macrophages isolated directly from growing mammary tumors, as well as activated peritoneal macrophages, were capable of inducing thioguanine resistance in line 66 cells. MH - Animal ; Catalase/METABOLISM ; *Drug Resistance ; Free Radicals ; Hypoxanthine Phosphoribosyltransferase/METABOLISM ; Macrophage Activation ; Macrophages/*PHYSIOLOGY ; Male ; Mammary Neoplasms, Experimental/*PHYSIOPATHOLOGY ; Mannitol/METABOLISM ; Mice ; Mutation ; Superoxide Dismutase/METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Thioguanine/ PHARMACODYNAMICS SO - Cancer Res 1986 May;46(5):2396-401 44 UI - 86184074 AU - Rehan A ; Wiggins RC ; Kunkel RG ; Till GO ; Johnson KJ TI - Glomerular injury and proteinuria in rats after intrarenal injection of cobra venom factor. Evidence for the role of neutrophil-derived oxygen free radicals. AB - The purpose of these studies was to determine how intravascular complement activation could lead to glomerular injury. Cobra venom factor (CVF) infused into the renal artery of rats resulted in increased excretion of protein in urine, which was maximal over the first 24 hours (51.2 +/- 6.0 mg/24 hours in CVF versus 14.1 +/- 0.9 mg/24 hours in saline-treated animals; P less than 0.001). Depletion of circulating neutrophils with anti-neutrophil serum significantly reduced the CVF-induced proteinuria in the first 24 hours (neutrophil depleted rats 22.7 +/- 2.8 mg/24 hours versus 63.4 +/- 9.9 mg/24 hours in neutrophil intact rats; P less than 0.005). Morphologic abnormalities (which were quantitated morphometrically) included accumulation of neutrophils in glomerular capillary loops, blebbing of endothelial cells, and epithelial cell foot process fusion. The increased protein excretion was reduced by 70% by simultaneous administration of catalase (23 +/- 4.3 mg/24 hours in CVF plus catalase versus 52.1 +/- 10 mg/24 hours in CVF alone; P less than 0.05). Catalase reduced glomerular endothelial cell blebbing and epithelial cell foot process fusion but not neutrophil accumulation in glomeruli as assessed by morphometry. In similar experiments superoxide dismutase, dimethyl sulfoxide, and deferoxamine did not prevent CVF-induced proteinuria. These studies, therefore, suggest that intravascular activation of complement in the rat causes glomerular injury and proteinuria which is dependent on neutrophils and upon the generation of hydrogen peroxide and/or its metabolites. MH - Animal ; Catalase/PHARMACODYNAMICS ; Cobra Venoms/ANTAGONISTS & INHIBITORS/*IMMUNOLOGY ; *Complement Activation/DRUG EFFECTS ; Deferoxamine/PHARMACODYNAMICS ; Dimethyl Sulfoxide/ PHARMACODYNAMICS ; Free Radicals ; Glomerular Filtration Rate ; Kidney Glomerulus/*PATHOLOGY/ULTRASTRUCTURE ; Male ; Microscopy, Electron ; Neutrophils/*METABOLISM ; Oxygen/METABOLISM ; Proteins/ ANALYSIS ; Proteinuria/*ETIOLOGY/PATHOLOGY ; Rats ; Rats, Inbred Strains ; Renal Artery ; Stains and Staining ; Superoxide Dismutase/PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Am J Pathol 1986 Apr;123(1):57-66 45 UI - 86170479 AU - Mizuno Y ; Ohta K TI - Regional distributions of thiobarbituric acid-reactive products, activities of enzymes regulating the metabolism of oxygen free radicals, and some of the related enzymes in adult and aged rat brains. AB - Regional distributions of thiobarbituric acid-reactive products, activities of enzymes regulating metabolism of oxygen free radicals, and some of the related enzymes were studied in 10 areas of adult and aged rat brains. Thiobarbituric acid-reactive products were lower in cerebral cortex, septal area, hippocampus, caudate-putamen, and substantia nigra compared with other areas studied in adult rats; however, they increased significantly in the former areas with aging. A slight but significant reduction in superoxide dismutase activity was noted in frontal cortex, septal area, caudate-putamen, and substantia nigra with aging. Glutathione peroxidase and reductase activities were highest in caudate-putamen and in substantia nigra. Glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase activities were lowest in cortical areas. Phosphofructokinase activity was lowest in septal area and hippocampus in aged rats. Glyceraldehyde-3-phosphate dehydrogenase activity showed only small regional and evolutional changes. Lactate dehydrogenase activity declined with age in most of the areas studied. sn-Glycerol-3-phosphate dehydrogenase activity showed small changes with aging except in hippocampus, where 40% reduction was noted. Generally, cerebral cortical areas, hippocampus, and septal areas were not particularly enriched in enzymes regulating the metabolism of oxygen free radicals. The results were discussed in relation to the role of free radicals in aging. MH - *Aging ; Animal ; Brain/*ENZYMOLOGY ; Brain Chemistry ; Comparative Study ; Free Radicals ; Glucosephosphate Dehydrogenase/METABOLISM ; Glutathione Peroxidase/METABOLISM ; Glutathione Reductase/METABOLISM ; Glyceraldehydephosphate Dehydrogenase/METABOLISM ; Glycerolphosphate Dehydrogenase/ METABOLISM ; *Glycolysis ; Indicators and Reagents ; Lactate Dehydrogenase/METABOLISM ; Male ; Oxygen/*METABOLISM ; Phosphofructokinase/METABOLISM ; Rats ; Superoxide Dismutase/ METABOLISM ; Support, Non-U.S. Gov't ; *Thiobarbiturates ; Tissue Distribution SO - J Neurochem 1986 May;46(5):1344-52 46 UI - 86145346 AU - James JL ; Friend DS ; MacDonald JR ; Smuckler EA TI - Alterations in hepatocyte plasma membrane in carbon tetrachloride poisoning. Freeze-fracture analysis of gap junction and electron spin resonance analysis of lipid fluidity. AB - The plasmalemma of the livers of rats treated with carbon tetrachloride (CCl4) were examined by freeze-fracture and electron spin resonance probe techniques. The rodents received by mouth either mineral oil alone (0 to 4.5 hours before sacrifice) or CCl4 in mineral oil (1:1) (2.5 ml of CCl4/kg, 0 to 3 hours before sacrifice). Rats were anesthetized with ether and livers were perfused in situ with saline either at ambient temperature or at 4 degrees C. After perfusion, livers were fixed in situ and processed for freeze-fracture and electron microscopy. Hepatocytes were isolated and incubated with 12-doxylstearic acid and subjected to electron spin resonance analysis. Electron microscopy revealed greater than a 2.5-fold increase in the individual mean gap junction size when rats were treated with mineral oil alone for 4.5 hours and the livers were processed at room temperature. The mean gap junction size in rats dosed with CCl4 for 0.5 hours before sacrifice equalled those of the group treated with mineral oil for 4.5 hours. Increases in gap junction size with CCl4 were progressive with time; by 3 hours, a 3.5-fold increase over controls was observed (p less than 0.05). When livers were perfused with iced saline, rats treated with mineral oil for 1.5 hours had a slight decrease (not significant) in mean gap junction size as compared to controls, while the size in rats treated for the same amount of time with CCl4 increased almost 5-fold over controls (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) MH - Animal ; Carbon Tetrachloride Poisoning/*PATHOLOGY ; Cell Membrane/DRUG EFFECTS/ULTRASTRUCTURE ; Cyclic N-Oxides/DIAGNOSTIC USE ; Electron Spin Resonance ; Freeze Fracturing ; Intercellular Junctions/DRUG EFFECTS/ULTRASTRUCTURE ; Liver/*DRUG EFFECTS/ ULTRASTRUCTURE ; Male ; Membrane Fluidity/*DRUG EFFECTS ; Mineral Oil/PHARMACODYNAMICS ; Rats ; Rats, Inbred Strains ; Spin Labels ; Support, U.S. Gov't, P.H.S. SO - Lab Invest 1986 Mar;54(3):268-74 47 UI - 86134895 AU - Burman WJ ; Martin WJ 2d TI - Oxidant-mediated ciliary dysfunction. Possible role in airway disease. AB - The effects of reactive species of oxygen on the airway are not well known. This study examined the effects of hydrogen peroxide (H2O2) on the structure and function of the airway epithelium. Tracheal rings were prepared from 200 g male rats. Damage to the airway epithelium was assayed by monitoring the ciliary beat frequency, the release of 51Cr, and histology. H2O2 at concentrations of 1.0 mM and above caused a very rapid decrease in ciliary beat frequency. After ten minutes' exposure to 1.0 mM, the ciliary beat frequency was 72 +/- 20 percent of control. Release of 51Cr was a less sensitive measure with significant release occurring after four hours of exposure to ciliotoxic concentrations of H2O2. Histologic changes were not evident within the experimental time period. All toxic effects of H2O2 were completely blocked by catalase. This study shows that H2O2 causes a rapid decline in ciliary activity and suggests that oxidant-mediated ciliary dysfunction could play a role in the pathogenesis of airway disease. The ciliary beat frequency provides a sensitive, physiologically relevant parameter for the in vitro study of these diseases. MH - Animal ; Chromium Radioisotopes/DIAGNOSTIC USE ; Cilia/DRUG EFFECTS/PATHOLOGY/PHYSIOLOGY ; Epithelium/PATHOLOGY/PHYSIOLOGY ; Free Radicals ; Hydrogen Peroxide/*TOXICITY ; Larynx/PATHOLOGY/ *PHYSIOPATHOLOGY ; Male ; Rats ; Rats, Inbred Strains ; Respiratory Tract Infections/PATHOLOGY/PHYSIOPATHOLOGY ; Time Factors ; Trachea/PATHOLOGY/*PHYSIOPATHOLOGY SO - Chest 1986 Mar;89(3):410-3 48 UI - 86133285 AU - Lindenschmidt RC ; Tryka AF ; Witschi HP TI - Inhibition of mouse lung tumor development by hyperoxia. AB - The hypothesis was tested that continuous hyperoxia would enhance the development of lung tumors in mice. In strain A/J mice treated with a single dose of urethan (1000 mg/kg) and exposed to 70% O2 for 16 wk, an average of 5 tumors per lung developed, whereas in animals kept in air, an average of 20 tumors per lung was found. When the animals were returned to air after oxygen exposure, it was found that a difference of 15 tumors per lung between the two groups persisted up to 1 yr later, indicating that O2 was tumoricidal. The shortest duration of O2 exposure to be effective was 4 wk, and delay of O2 exposure up to 12 wk after urethan still was effective in reducing the number of developing tumors. Histopathology showed that continued exposure to 70% O2 produced some hyperplasia of the bronchiolar epithelium and only very discrete changes in the pulmonary parenchyma. Analysis of cell proliferation patterns with a continuous [3H]thymidine labeling technique showed a persistent high cell labeling in the bronchiolar epithelium and a temporary increase in alveolar wall cell labeling. Chronic hyperoxia failed to alter the activities of pulmonary superoxide dismutase or glucose-6-phosphate dehydrogenase. Ornithine decarboxylase, on the other hand, was increased as long as the animals remained exposed to oxygen. It was concluded that hyperoxia kills developing tumor cells in mouse lung. MH - Animal ; Body Weight ; Cell Division ; Dose-Response Relationship, Drug ; Free Radicals ; Lung/ENZYMOLOGY/PATHOLOGY ; Lung Neoplasms/PATHOLOGY/*PREVENTION & CONTROL ; Male ; Mice ; Mice, Inbred Strains ; Ornithine Decarboxylase/ANALYSIS ; Oxygen/ *PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Thymidine/METABOLISM ; Time Factors ; Tritium/ DIAGNOSTIC USE ; Urethane/TOXICITY SO - Cancer Res 1986 Apr;46(4 Pt 2):1994-2000 49 UI - 86132682 AU - Pellmar T TI - Electrophysiological correlates of peroxide damage in guinea pig hippocampus in vitro. AB - To study the effects of active oxygen on neuronal electrophysiology, hippocampal brain slices were exposed to hydrogen peroxide plus ferrous sulfate which react to produce hydroxyl free radicals. Analysis of extracellularly recorded somatic and dendritic responses to orthodromic stimulation indicated a decrease in both synaptic efficacy and impairment of action potential generation. MH - Animal ; Brain Diseases/CHEMICALLY INDUCED ; Cerebral Ischemia/ PHYSIOPATHOLOGY ; Disease Models, Animal ; Drug Interactions ; Evoked Potentials/DRUG EFFECTS ; Ferrous Compounds/*TOXICITY ; Free Radicals ; Guinea Pigs ; *Hippocampus ; Hydrogen Peroxide/ *TOXICITY ; In Vitro ; Iron/*TOXICITY ; Lipid Peroxides/ BIOSYNTHESIS ; Male ; Support, U.S. Gov't, Non-P.H.S. SO - Brain Res 1986 Feb 5;364(2):377-81 50 UI - 86127952 AU - Guice KS ; Miller DE ; Oldham KT ; Townsend CM Jr ; Thompson JC TI - Superoxide dismutase and catalase: a possible role in established pancreatitis. AB - The mechanism of cerulein-induced acute pancreatitis may involve the production of free radicals in excess of the capacity of endogenous intracellular scavengers. These radicals destroy the cellular membranes, releasing digestive enzymes and cellular proteins into the interstitium. Thereafter, a cascade of events, including polymorphonuclear infiltration and complement activation, leads to pancreatic destruction. The present study demonstrates that superoxide dismutase and catalase reduce the ultrastructural and biochemical injury associated with cerulein-induced acute pancreatitis in rats. Pretreatment with superoxide dismutase and catalase 30 minutes before injury did not appear to be protective, presumably because the half-life of intravenous superoxide dismutase is only 6 minutes. This and similar studies suggest a potential clinical role for free radical scavengers in acute established pancreatitis. MH - Acute Disease ; Animal ; Caerulein ; Catalase/*PHARMACODYNAMICS ; Comparative Study ; Disease Models, Animal ; DNA/METABOLISM ; Free Radicals ; Human ; In Vitro ; Male ; Pancreas/METABOLISM/ ULTRASTRUCTURE ; Pancreatitis/CHEMICALLY INDUCED/*METABOLISM/ PATHOLOGY ; Rats ; Rats, Inbred Strains ; RNA/METABOLISM ; Superoxide Dismutase/*PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Am J Surg 1986 Jan;151(1):163-9 51 UI - 86124048 AU - Green CJ ; Healing G ; Lunec J ; Fuller BJ ; Simpkin S TI - Evidence of free-radical-induced damage in rabbit kidneys after simple hypothermic preservation and autotransplantation. AB - Rabbit kidneys were stored for 24 or 48 hr at 0 degree C after single-passage vascular flush with 30 ml of cold hypertonic citrate solution or 0.9% isotonic sodium chloride solution. They were then subjected to in vitro biochemical assay for evidence of free-radical damage immediately after storage or after they had been orthotopically autotransplanted and reperfused with blood in vivo for 60 min. Kidney homogenates were incubated at 37 degrees C and assayed for fluorescent conjugated Schiff bases as indicators of lipid peroxidation, as well as for superoxide dismutase activity and reduced and oxidized glutathione. In kidneys flushed with hypertonic citrate, no evidence of peroxidation could be detected immediately after storage for 24 or 48 hr. However, after in vivo reperfusion significantly more peroxidation (P less than 0.01) was evident. Storage in isotonic saline solution produced still higher levels of peroxidation damage whether reperfused or not (P less than 0.001). Schiff base formation was inversely proportional to the reduced and oxidized glutathione levels measured. No changes in superoxide dismutase levels could be detected. It is concluded that lipid peroxidation is important during cold ischemia but most damage occurs during the 60-min of reperfusion in vivo immediately after transplantation. MH - Animal ; Citrates ; Free Radicals ; Glutathione/METABOLISM ; Hypertonic Solutions ; *Hypothermia, Induced/ADVERSE EFFECTS ; Kidney/METABOLISM/*PATHOLOGY ; Lipid Peroxides/*METABOLISM/ TOXICITY ; *Organ Preservation ; Oxidation-Reduction ; Rabbits ; Schiff Bases ; Superoxide Dismutase/METABOLISM ; Support, Non-U.S. Gov't ; Transplantation, Autologous SO - Transplantation 1986 Feb;41(2):161-5 52 UI - 86123470 AU - Manson PN ; Narayan KK ; Im MJ ; Bulkley GB ; Hoopes JE TI - Improved survival in free skin flap transfers in rats. AB - We have demonstrated previously that oxygen-derived free radicals are important mediators of tissue injury in experimental island skin flaps that have been subjected to prolonged ischemia (vascular occlusion) followed by reperfusion. In this study the role of oxygen free radicals in ischemia/reperfusion injury has been investigated in free flap transfers. Groin skin flaps were harvested, stored at room temperature for 21 to 24 hours, and transplanted to the contralateral groin. These free flap transfers normally exhibit a high incidence of complete necrosis. Treatment before the onset of reperfusion with a single dose of superoxide dismutase (SOD), a scavenger of superoxide radicals, increased the survival rate of these skin flaps from 38% in the control group to 76% (p less than 0.025). Tissue levels of SOD were measured before ischemia, after ischemia but before reperfusion, and 30 minutes after reperfusion: untreated flap tissues, which were destined to undergo necrosis, exhibited a significant decrease in SOD activity after reperfusion, whereas SOD-treated flap tissues, destined to survive, demonstrated increased enzyme activity. High levels of tissue SOD activity thus appeared to be associated with improved flap survival. The results have significant clinical implications with regard to organ preservation and transplantation. MH - Animal ; Female ; Free Radicals ; Ischemia ; Necrosis ; Perfusion ; Rats ; Rats, Inbred Strains ; Skin/BLOOD SUPPLY/ENZYMOLOGY/ PATHOLOGY/*TRANSPLANTATION ; Superoxide Dismutase/METABOLISM/ PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; *Surgical Flaps ; Tissue Preservation SO - Surgery 1986 Feb;99(2):211-5 53 UI - 86116883 AU - Otani H ; Engelman RM ; Rousou JA ; Breyer RH ; Lemeshow S ; Das DK TI - Cardiac performance during reperfusion improved by pretreatment with oxygen free-radical scavengers. AB - We studied the effects of oxygen free radicals on cardiac performance during reperfusion of ischemic myocardium. The pig heart, isolated in situ, was subjected to 60 minutes of regional ischemia at normothermia by occlusion of the left anterior descending coronary artery followed by 60 minutes of hypothermic cardioplegic arrest and 60 minutes of normothermic reperfusion. The oxygen free-radical scavengers, superoxide dismutase and catalase, were administered before occlusion of the left anterior descending coronary artery in the experimental group. The generation of free radicals in the untreated group, estimated by the measurement of malondialdehyde in the perfusate, was significant during reperfusion and was associated with a corresponding increase in creatine kinase. Superoxide dismutase and catalase significantly slowed the appearance of malondialdehyde and the release of creatine kinase during reperfusion. Superoxide dismutase and catalase did not alter coronary flow and myocardial oxygen extraction or consumption during occlusion of the left anterior descending coronary artery; however, coronary flow and oxygen consumption were significantly higher (p less than 0.05) during reperfusion in hearts treated with antioxidants. Left ventricular developed pressure and its maximum first derivative were measured under isovolumic conditions. In the untreated group, left ventricular developed pressure and its maximum first derivative declined to 61.1% and 57.1% of baseline values, respectively, after 60 minutes' occlusion of the left anterior descending, and to 45% of baseline values after 15 minutes of reperfusion. The decline in left ventricular developed pressure and its maximum first derivative during reperfusion was significantly (p less than 0.05) inhibited by superoxide dismutase and catalase, but left ventricular end-diastolic pressure was not significantly altered. These results implicate oxygen-derived free radicals in the injury resulting from reperfusion of ischemic myocardium and suggest that oxygen free-radical scavengers effectively protect against such injury. MH - Animal ; Blood Flow Velocity/DRUG EFFECTS ; Catalase/ PHARMACODYNAMICS ; Coronary Disease/ENZYMOLOGY/METABOLISM/ *PHYSIOPATHOLOGY ; Creatine Kinase/METABOLISM ; Female ; Free Radicals ; Male ; Malondialdehyde/METABOLISM ; *Myocardial Revascularization/ADVERSE EFFECTS/METHODS ; Myocardium/METABOLISM ; Oxygen/METABOLISM/*PHARMACODYNAMICS ; Oxygen Consumption/DRUG EFFECTS ; Perfusion/ADVERSE EFFECTS ; Potassium ; Premedication ; Superoxide Dismutase/PHARMACODYNAMICS ; Support, U.S. Gov't, P.H.S. ; Swine SO - J Thorac Cardiovasc Surg 1986 Feb;91(2):290-5 54 UI - 86103436 AU - Lefer AM TI - Leukotrienes as mediators of ischemia and shock. AB - Leukotrienes have been implicated as mediators of ischemia and shock. Recent evidence has been obtained supporting the four major criteria of acceptance of leukotrienes as mediators of shock, namely (a) increased concentration in body fluids during shock states, (b) ability to exert significant pathophysiologic effects which aggravate ischemia and shock, (c) amelioration of the shock state by leukotriene synthesis inhibitors and leukotriene receptor antagonists, and (d) production of a shock-like state by exogenous administration of leukotrienes. In conclusion, both LTB4 and the peptide leukotrienes (e.g. LTC4, LTD4 and LTE4) also known as the slow reacting substance of anaphylaxis (SRS-A) can be considered as mediators of ischemia and shock. Although difficulties exist with measuring leukotrienes in circulating blood and in obtaining long lasting selective blockers of leukotriene synthesis, innovative experiments measuring leukotrienes in bile and other body fluids and in employing specific leukotriene receptor antagonists have helped in assessing the significance of the leukotrienes in shock states. Additional studies are necessary to evaluate these findings in perspective, and to compare and contrast the role of leukotrienes to that of other vascular mediators including prostaglandins and thromboxanes, as well as non-eicosanoids including serotonin, histamine, angiotensin II and vasopressin, all of which can play a mediator role in ischemia and shock states. Further clarification of these issues promises to open exciting new chapters in shock research. MH - Animal ; Free Radicals ; Human ; Ischemia/*PHYSIOPATHOLOGY ; Leukotrienes B/*PHYSIOLOGY ; Lipoxygenases/ANTAGONISTS & INHIBITORS ; Receptors, Immunologic/DRUG EFFECTS ; Receptors, Prostaglandin/DRUG EFFECTS ; Shock/*PHYSIOPATHOLOGY ; Support, U.S. Gov't, P.H.S. ; SRS-A/*PHYSIOLOGY SO - Biochem Pharmacol 1986 Jan 15;35(2):123-7 55 UI - 86082395 AU - Till GO ; Ward PA TI - Systemic complement activation and acute lung injury. AB - Experimental studies of rats have provided significant evidence that intravascular complement activation after i.v. injection of cobra venom factor (CVF) or thermal injury of skin can result in acute lung injury. This has been determined by morphological changes in lung and increases in lung vascular permeability. Systemic complement activation is associated with an early appearance of C5-derived chemotactic activity in the circulation coincident with the development of transient neutropenia, followed by extensive granulocytosis and sequestration of neutrophils in lung interstitial capillaries. The acute pulmonary injury depends on availability of complement and neutrophils. Depletion of either complement or blood neutrophils before CVF injection or thermal injury will prevent development of lung injury. Interventional studies with catalase, scavengers of hydroxyl radical OH., and iron chelators have revealed that the acute pulmonary injury is related to production of oxygen-derived free radicals by activated neutrophils. OH. appears to be the key mediator involved in the acute lung microvascular injury. MH - Acute Disease ; Animal ; Capillary Permeability/DRUG EFFECTS ; Cobra Venoms/PHARMACODYNAMICS ; *Complement Activation/DRUG EFFECTS ; Complement 5/PHYSIOLOGY ; Endothelium/PATHOLOGY ; Free Radicals ; Heat ; Inflammation/PATHOLOGY/PHYSIOPATHOLOGY ; Leukocyte Count ; Lung/BLOOD SUPPLY/*PATHOLOGY ; Neutrophils/ METABOLISM/*PATHOLOGY ; Rats ; Support, U.S. Gov't, P.H.S. SO - Fed Proc 1986 Jan;45(1):13-8 1 UI - 87102360 AU - Kubow S ; Bray TM ; Bettger WJ TI - Effects of dietary zinc and copper on free radical production in rat lung and liver. AB - The effects of dietary copper and zinc on free radical production in lung and liver microsomes were studied in male weanling rats. The rats were fed for 6 weeks on one of seven diets, with different copper and zinc concentrations representing low, adequate, and high dietary levels of copper and low and adequate levels of zinc. Rats were put on diets arranged in a 3 X 2 factorial design with copper and zinc supplementations of 0, 15, and 500 mg/kg and 0.5 or 100 mg/kg, respectively. The low copper diet depressed copper levels in both the lungs and liver, although zinc levels were unchanged in rats on the low zinc diets. Endogenous carbon-centered lipid radical production in microsomes induced by NADPH was measured using spin-trapping techniques. The low zinc diets increased free radical production in lung microsomes but not in liver microsomes. No change in free radical production was observed in lung or liver microsomes obtained from rats on low copper diets. The data indicate that endogenous free radical production is increased in lung microsomes as a function of dietary zinc deficiency but is not influenced by copper status. MH - Animal ; Copper/*PHARMACODYNAMICS ; Diet ; Free Radicals ; Liver/ *METABOLISM ; Lung/*METABOLISM ; Male ; Microsomes/ENZYMOLOGY ; NADP/METABOLISM ; Organ Weight/DRUG EFFECTS ; Rats ; Rats, Inbred Strains ; Zinc/*PHARMACODYNAMICS SO - Can J Physiol Pharmacol 1986 Oct;64(10):1281-5 2 UI - 87099822 AU - Rice-Evans C ; Omorphos SC ; Baysal E TI - Sickle cell membranes and oxidative damage. AB - Sickle erythrocytes and their membranes are susceptible to endogenous free-radical-mediated oxidative damage which correlates with the proportion of irreversibly sickled cells. The suppression of incubation-induced oxidative stress by antioxidants, free radical scavengers and an iron chelator suggest that oxidation products of membrane-bound haemoglobin contribute towards the pathology of the disease. MH - Anemia, Sickle Cell/*BLOOD ; Antioxidants/PHARMACODYNAMICS ; Ascorbic Acid/PHARMACODYNAMICS ; Deferoxamine/PHARMACODYNAMICS ; Erythrocyte Membrane/DRUG EFFECTS/*METABOLISM ; Erythrocytes, Abnormal/DRUG EFFECTS/*METABOLISM ; Free Radicals ; Human ; In Vitro ; Iron/PHARMACODYNAMICS ; Oxidation-Reduction ; Support, Non-U.S. Gov't SO - Biochem J 1986 Jul 1;237(1):265-9 3 UI - 87039059 AU - DeCaprio AP TI - Mechanisms of in vitro pyrrole adduct autoxidation in 2,5-hexanedione-treated protein. AB - The neurotoxic gamma-diketone, 2,5-hexanedione, reacts with axonal protein amine residues to form 2,5-dimethylpyrrole adducts. Current evidence implicates this reaction as the potentially critical step in gamma-diketone neurotoxicity, although it is unclear whether pyrrole formation per se is sufficient to induce neuropathy or whether secondary autoxidative reactions are also required. The present in vitro study examines aspects of pyrrole formation and the secondary phenomena of chromophore development and covalent protein crosslinking in 2,5-hexanedione-treated protein. p-Dimethylaminobenzaldehyde (DMAB)-detectable pyrrole concentrations decreased linearly with time when pyrrolylated bovine serum albumin (pyrrole-BSA) was incubated under air, but remained unchanged following N2 incubation. The air-induced decrease was accompanied by the appearance of chromophores and crosslinked protein. Covalent crosslinking of pyrrole-BSA was pH-dependent, with relatively increased intermolecular bridging at pH 7.4 as compared to pH 9.5. Chromophore formation and the loss in DMAB-detectable pyrrole were also accelerated at the lower pH. Autoxidative parameters were inhibited in the presence of a free radical scavenger (ascorbic acid) but induced by free radical initiators (potassium persulfate and 2,2'-azobis[2-amidinopropane hydrochloride]). In vitro incubation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of combinations of bovine serum albumin, ribonuclease, pyrrole-BSA, and pyrrolylated ribonuclease revealed that the intermolecular crosslinking pathway was mediated by pyrrole-pyrrole bridging. These findings demonstrate that the secondary autoxidative phenomena following pyrrole adduct formation in gamma-diketone-treated protein proceed via pH-dependent, free radical-mediated mechanisms. If similar mechanisms are present in vivo, the results also suggest that intermolecular covalent crosslinking of pyrrolylated axonal protein may be less widespread and more specific than previously thought. MH - Animal ; Ascorbic Acid/PHARMACODYNAMICS ; Cattle ; Free Radicals ; Hexanones/*PHARMACODYNAMICS ; Hydrogen-Ion Concentration ; Intermediate Filaments/METABOLISM ; Ketones/*PHARMACODYNAMICS ; Oxidation-Reduction/DRUG EFFECTS ; Pyrroles/*METABOLISM ; Serum Albumin, Bovine/METABOLISM ; Support, U.S. Gov't, P.H.S. SO - Mol Pharmacol 1986 Nov;30(5):452-8 4 UI - 87030920 AU - Tan KH ; Meyer DJ ; Coles B ; Ketterer B TI - Thymine hydroperoxide, a substrate for rat Se-dependent glutathione peroxidase and glutathione transferase isoenzymes. AB - The thymine hydroperoxide, 5-hydroperoxymethyluracil, is a substrate for Se-dependent glutathione (GSH) peroxidase and the Se-independent GSH peroxidase activity associated with the GSH transferase fraction. These enzymes may contribute to repair mechanisms for damage caused by oxygen radicals. GSH transferases 1-1, 2-2, 3-3, 4-4, 6-6, and 7-7 [(1984) Biochem. Pharmacol. 33, 2539-2540] are shown to differ considerably in their ability to utilize this substrate. For example, high activity is found in GSH transferase 6-6 which is the major isoenzyme in spermatogenic tubules where DNA synthesis is so active and faithful DNA replication so important. The activity of the purified GSH transferase isoenzymes towards 5-hydroperoxymethyluracil is comparable with their activity towards other endogenous substrates related to cellular peroxidation such as linoleate hydroperoxide and 4-hydroxynon-2-enal or biologically important xenobiotic metabolites such as benzo(a)pyrene-7,8-diol-9,10-oxide. MH - Animal ; DNA Repair ; Free Radicals ; Glutathione Peroxidase/ *METABOLISM ; Glutathione Transferases/*METABOLISM ; Isoenzymes/ *METABOLISM ; Oxygen ; Rats ; Selenium/*PHARMACODYNAMICS ; Substrate Specificity ; Support, Non-U.S. Gov't ; Thymine/ *ANALOGS & DERIVATIVES/METABOLISM SO - FEBS Lett 1986 Oct 27;207(2):231-3 5 UI - 87025777 AU - Goshima N ; Wadano A ; Miura K TI - 3-Hydroxykynurenine as O2-. scavenger in the blowfly, Aldrichina grahami. AB - We studied the distribution of O2-.-scavenging activity in 6-day-old larvae of Aldrichina grahami. Total activity was highest in the muscle. The specific activity per milligram of protein in the Malpighian tubules was highest, 10 times the highest elsewhere. Most of the O2-. scavenging activity in muscle depended on superoxide dismutase. However, the activity in the Malpighian tubules mostly depended on 3-hydroxykynurenine. MH - Animal ; Ascorbic Acid/ANALYSIS ; Chromatography, High Pressure Liquid ; Diptera/*METABOLISM ; Free Radicals ; Kynurenine/ *ANALOGS & DERIVATIVES/METABOLISM ; Molecular Weight ; Oxygen/ *METABOLISM ; Tissue Distribution ; Uric Acid/ANALYSIS SO - Biochem Biophys Res Commun 1986 Sep 14;139(2):666-72 6 UI - 87025745 AU - Chan TM ; Chen E ; Tatoyan A ; Shargill NS ; Pleta M ; Hochstein P TI - Stimulation of tyrosine-specific protein phosphorylation in the rat liver plasma membrane by oxygen radicals. AB - Incorporation of 32P from [gamma-32P]ATP into endogenous proteins, added histone and the copolymers Glu 80 Tyr 20 by rat liver plasma membranes was markedly increased by several naphthoquinones, including menadione. This stimulation was most marked with Glu 80 Tyr 20, has an absolute requirement for either dithiothreitol or reduced glutathione, and was inhibited by superoxide dismutase, catalase, and desferrioxamine to varying degrees depending on the quinones used. Their effectiveness in stimulating the apparent tyrosine-specific protein phosphorylation correlated with the rates of DTT-dependent redox cycling measured by oxygen consumption. Increased protein phosphorylation was also seen with particulate fractions isolated from hepatocytes incubated with quinones. A free radical-mediated mechanism is suggested for the quinone stimulation of protein phosphorylation. MH - Adenosine Triphosphate/METABOLISM ; Animal ; Cell Membrane/DRUG EFFECTS/*ENZYMOLOGY ; Deferoxamine/PHARMACODYNAMICS ; Free Radicals ; Hydroquinones/PHARMACODYNAMICS ; Liver/*CYTOLOGY ; Naphthoquinones/METABOLISM/PHARMACODYNAMICS ; Oxidation-Reduction ; Oxygen Consumption/DRUG EFFECTS ; Oxygen/*PHARMACODYNAMICS ; Protein-Tyrosine Kinase/*METABOLISM ; Quinones/PHARMACODYNAMICS ; Rats ; Structure-Activity Relationship ; Superoxide Dismutase/ METABOLISM ; Support, U.S. Gov't, P.H.S. ; Vitamin K/ PHARMACODYNAMICS SO - Biochem Biophys Res Commun 1986 Sep 14;139(2):439-45 7 UI - 87001950 AU - Hennekens CH ; Mayrent SL ; Willett W TI - Vitamin A, carotenoids, and retinoids. AB - One promising area of current research in chemoprevention is the possibility that micronutrients, including vitamin A analogues, may decrease cancer incidence. The term "vitamin A: refers either to retinol (preformed vitamin A) and its synthetic analogues, or to certain carotenoids (provitamin A), which are converted to retinol in the body, as needed. Retinol and the other retinoids are integrally involved in cell growth and differentiation, which may affect carcinogenesis. Such a role has been supported by a large number of in vitro and animal experiments. Data from studies among humans are sparse, in part because most dietary studies have been conducted in populations in which the vast majority of vitamin A intake is actually beta-carotene and other carotenoids, found in carrots and other vegetables and fruits. Although the carotenoids do not have the hormone-like properties of retinol, they do have a potent antioxidant effect and could thus reduce cancer risk by preventing tissue damage due to oxidation. This possibility is supported by data from a large number of observational epidemiologic studies, most of which have consistently found an inverse relation between consumption of carotene-rich vegetables and cancer risk. However, the only direct way to determine whether carotenoids have a beneficial effect is through large, carefully conducted randomized trials. Several such studies are currently underway and should provide sound evidence on which future medical policy and practice can be based. MH - Carotenoids/*PHYSIOLOGY ; Clinical Trials ; Free Radicals ; Human ; Neoplasms/ETIOLOGY/*PREVENTION & CONTROL ; Random Allocation ; Retinoids/*PHYSIOLOGY ; Vitamin A/*PHYSIOLOGY SO - Cancer 1986 Oct 15;58(8 Suppl):1837-41 8 UI - 87000763 AU - Cadet J ; Berger M ; Decarroz C ; Wagner JR ; van Lier JE ; Ginot YM ; Vigny P TI - Photosensitized reactions of nucleic acids. AB - The main effects of near-ultraviolet and visible light on cellular DNA are reviewed with emphasis on base lesions, oligonucleotide single-strand breaks and DNA-protein cross-links. Model system photosensitization reactions of DNA are also discussed. This includes photodynamic effects, menadione-mediated photooxidation, photoionization of antibiotics, the photochemistry of 5-halogenopyrimidines and urocanic acid. MH - p-Aminobenzoic Acid ; Animal ; Antineoplastic Agents ; Bacillus Subtilis/GENETICS ; Bromouracil ; Chemistry ; Daunorubicin/ RADIATION EFFECTS ; Doxorubicin/RADIATION EFFECTS ; DNA Repair ; DNA/*RADIATION EFFECTS ; DNA, Bacterial/RADIATION EFFECTS ; Escherichia Coli/GENETICS ; Free Radicals ; Guanine ; Hematoporphyrins ; Human ; Light ; Nucleosides ; Oxidation-Reduction ; Oxygen/PHARMACODYNAMICS ; Photochemistry ; Pyrimidine Dimers/RADIATION EFFECTS ; Pyrimidines ; Review ; Rose Bengal ; Support, Non-U.S. Gov't ; Ultraviolet Rays ; Uridine/ ANALOGS & DERIVATIVES ; Urocanic Acid ; Vitamin K SO - Biochimie 1986 Jun;68(6):813-34 9 UI - 86295796 AU - Cataldi de Flombaum MA ; Stoppani AO TI - Inactivation of mitochondrial adenosine triphosphatase from Trypanosoma cruzi by oxygen radicals. AB - Incubation of Trypanosoma cruzi mitochondrial ATPase (Fo-F1) with the xanthine oxidase system (XO), Fenton's reagent (Fe2+ + H2O2) and the ascorbate-Cu system, caused gradual loss of enzyme activity, which increased as a function of incubation time and rate of oxygen radical generation. The essential role of OH. radicals for ATPase inactivation was supported by a) the enzyme protection afforded by superoxide dismutase, catalase and mannitol, when using the XO system; b) the similar effect of mannitol and benzoate with Fenton's reagent; c) the similar effect of catalase, EDTA and histidine with the ascorbate-Cu system; d) the increased rate of ATPase inactivation by 1) the XO system supplemented with chelated iron, and 2) the ascorbate-Cu system supplemented with H2O2. Comparison of oxygen radical generators for their action on membrane-bound (Fo-F1) and soluble F1 revealed that ascorbate-Cu was the most effective one, possibly because of its capability of producing OH. radicals that react preferentially with the enzyme at their formation site. MH - Adenosine Triphosphatase/*ANTAGONISTS & INHIBITORS ; Animal ; Ascorbic Acid/PHARMACODYNAMICS ; Catalase/METABOLISM ; Cattle ; Copper/PHARMACODYNAMICS ; Erythrocytes/ENZYMOLOGY ; Free Radicals ; Hydroxides/*PHARMACODYNAMICS ; Kinetics ; Liver/ENZYMOLOGY ; Mitochondria/*ENZYMOLOGY ; Superoxide Dismutase/BLOOD ; Support, Non-U.S. Gov't ; Trypanosoma Cruzi/*ENZYMOLOGY ; Xanthine Oxidase/ METABOLISM SO - Biochem Int 1986 Jun;12(6):785-93 10 UI - 86133694 AU - Moreno SN ; Docampo R TI - Reduction of the metallochromic indicators murexide and tetramethylmurexide to their free radical metabolites by cytoplasmic enzymes and reducing agents. AB - Murexide underwent reduction by rat liver cytosolic fraction or a hypoxanthine-xanthine oxidase system to produce a free radical metabolite. Reduction of murexide by the freshly prepared cytosolic fraction depended upon the presence of ascorbic acid. N1-Methylnicotinamide, xanthine or hypoxanthine, in that order, could also serve as a source of reducing equivalents for the production of that free radical by the cytosolic fraction. Several thiol compounds (GSH, cysteine, and cysteamine), pyridine nucleotides (NADH, NADPH) and ascorbic acid were also effective in generating the murexide-derived free radical. Tetramethyl murexide was also reduced to its free radical derivative by a hypoxanthine-xanthine oxidase system. MH - Animal ; Ascorbic Acid/PHARMACODYNAMICS ; Barbiturates/ *METABOLISM ; Cysteine/PHARMACODYNAMICS ; Cytosol/ENZYMOLOGY ; Electron Spin Resonance ; Free Radicals ; Glutathione/ PHARMACODYNAMICS ; Liver/*ENZYMOLOGY ; Male ; Murexide/ANALOGS & DERIVATIVES/*METABOLISM ; Oxidation-Reduction/*DRUG EFFECTS ; Oxygen Consumption/DRUG EFFECTS ; Rats ; Support, U.S. Gov't, P.H.S. ; Xanthine Oxidase/*METABOLISM SO - Chem Biol Interact 1986 Jan;57(1):17-25 11 UI - 86104725 AU - Helgestad J ; Storm-Mathisen I ; Lie SO TI - Vitamin C and thiol reagents promote the in vitro growth of murine granulocyte/macrophage progenitor cells by neutralizing endogenous inhibitor(s). AB - Growth of murine hemopoietic cells in culture requires the presence of a stimulator of stem cell proliferation, "colony stimulating factor: (CSF). A widely used source of CSF is lung conditioned medium (LCM). We have earlier shown that the great variability of CSF activities in different batches of LCM is due to varying amounts of inhibitor(s). The present study expands the observation that the addition of ascorbic acid to the murine bone marrow soft agar assay system removes the inhibitory activity. The vitamin probably acts as an antioxidant or free radical scavenger, since addition of reduced (but not oxidized) glutathione, cysteine, dithiothreitol or 2-mercaptoethanol to the cultures also inactivates the endogeneous inhibitor. Cysteine and glutathione gave the highest colony numbers, were active at concentrations present in body fluids and did not inhibit colony growth even at concentrations ten times higher than optimum. No synergistic effects could be observed between the different antioxidants. At optimum concentration (usually 0.45 mmol/l) the otherwise bell-shaped dose-response curve for conditioned medium changed to a sigmoid curve. Antioxidants had no growth promoting effect in the absence of CSF. The presence of cysteine or vitamin C revealed CSF-like activity in conditioned media of tissues not considered to be potent producers of such factors. It has been reported that individual batches of foetal calf serum contain different levels of reduced glutathione, and we suggest that one of the batch variable growth regulators in foetal calf serum may be reduced glutathione. The results indicate a possible physiological role of antioxidants in granulopoiesis and suggest that cysteine or reduced glutathione should be freshly added to culture systems assaying CSF and/or granulocyte macrophage progenitor cells. MH - Animal ; Antioxidants/PHARMACODYNAMICS ; Ascorbic Acid/ *PHARMACODYNAMICS ; Cells, Cultured ; Colony-Forming Units Assay ; Colony-Stimulating Factor/ANALYSIS ; Comparative Study ; Culture Media ; Cysteine/*PHARMACODYNAMICS ; Dithiothreitol/ PHARMACODYNAMICS ; Free Radicals ; Glutathione/*PHARMACODYNAMICS ; Granulocytes ; *Hematopoiesis ; Hematopoietic Stem Cells/*DRUG EFFECTS ; Macrophages ; Male ; Mercaptoethanol/PHARMACODYNAMICS ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Proteins/ ANALYSIS ; Support, Non-U.S. Gov't SO - Blut 1986 Jan;52(1):1-8 12 UI - 86323081 AU - Marx G ; Chevion M TI - Site-specific modification of albumin by free radicals. Reaction with copper(II) and ascorbate. AB - Exposure of albumin to Cu(II) (10-100 microM) and ascorbate (0.1-2 mM) results in extensive molecular modifications, indicated by decreased fluorescence and chain breaks. The rate of utilization of molecular oxygen and ascorbate as a function of Cu(II) concentration is non-linear at copper/albumin ratios of greater than 1. It appears that Cu(II) bound to the tightest albumin-binding site is less available to the ascorbate than the more loosely bound cation. SDS/polyacrylamide-gel electrophoresis reveals new protein bands corresponding to 50, 47, 22, 18 and 3 kDa. For such a cleavage pattern, relatively few (approximately 3) and rather specific chain breaks occurred. Repeated addition of portions of ascorbate to the albumin/Cu(II) mixture results in increased intensity of the new bands. The absence of Cu(II) or the presence of metal chelating agents is inhibitory. There was no evidence of intermolecular cross-linking or of the formation of insoluble, albumin-derived, material. A mechanism is proposed wherein the loosely bound Cu(II) participates in a Fenton-type reaction. This generates OH. radicals, which rapidly inter-react with the protein and modify it in a 'site-specific' manner. MH - Ascorbic Acid/*METABOLISM ; Binding Sites ; Copper/*METABOLISM ; Electrophoresis, Polyacrylamide Gel ; Free Radicals ; Human ; Oxidation-Reduction ; Serum Albumin/*METABOLISM ; Spectrophotometry ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. SO - Biochem J 1986 Jun 1;236(2):397-400 13 UI - 86311644 AU - Mizoi K ; Suzuki J ; Imaizumi S ; Yoshimoto T TI - Development of new cerebral protective agents: the free radical scavengers. AB - The generation of the free radical reaction in the ischaemic and hypoxic brain has been demonstrated using the chemiluminescence and the ESR techniques. The effects of various drugs, thought to be free radical scavengers were then tested in the ischaemic brain of dogs. It was found that not only mannitol, but also vitamin E, dexamethasone and other drugs have significant cerebral protective effects--particularly when administered together. Moreover, remarkable improvements were found when these 3 drugs were administered with the artificial blood substitute, PFC, which is known to have a high oxygen-carrying capacity. Finally, using the chemiluminescence and the ESR method, we have demonstrated that mannitol, vitamin E and glucocorticoids act as free radical scavengers and particularly mannitol acts as a scavenger of the hydroxy radical. MH - Animal ; Anoxia/*DRUG THERAPY/METABOLISM ; Cerebral Ischemia/ *DRUG THERAPY/METABOLISM ; Dexamethasone/*THERAPEUTIC USE ; Disease Models, Animal ; Dogs ; Electroencephalography ; Electron Spin Resonance ; Free Radicals ; Luminescence ; Male ; Mannitol/ *THERAPEUTIC USE ; Rats ; Rats, Inbred Strains ; Vitamin E/ *THERAPEUTIC USE SO - Neurol Res 1986 Jun;8(2):75-80 14 UI - 86309261 AU - Cavarocchi NC ; England MD ; O'Brien JF ; Solis E ; Russo P ; Schaff HV ; Orszulak TA ; Pluth JR ; Kaye MP TI - Superoxide generation during cardiopulmonary bypass: is there a role for vitamin E? AB - The cytotoxic metabolites of oxygen [superoxide (O-2), hydrogen peroxide (H2O2), and hydroxyl (OH.)] have been demonstrated to be involved in the peroxidation of membrane lipids consequently altering membrane composition, morphology, and function. Of all the lines of defense adopted by living organisms against toxic oxygen free radicals, vitamin E is most effective in the prevention of membrane damage. Cardiopulmonary bypass (CPB) has been shown to activate complement and cause sequestration of leukocytes which can recruit, adhere, and stimulate release of cytotoxic oxygen radicals. A prospective study of 30 patients evaluated the effects of CPB with and without an exogenous free radical scavenger (Group I, N = 20, control) and (Group II, N = 10, vitamin E) on H2O2 (a marker of oxygen free radicals) malonaldehyde (a marker of lipid peroxidation), transpulmonary leukosequestration, and plasma levels of vitamins E and C. Group I showed a progressive increase in H2O2 during CPB from 65 +/- 6 to 130 +/- 11 micron/ml (P less than 0.0001); plasma vitamin E decreased from 15 +/- 3 to 6 +/- 1 mg/liter (P less than 0.0001) while vitamin C increased from 1.6 +/- .3 to 2.3 +/- .3 mg/dl (P less than 0.0001). Group II showed no significant increase in H2O2 (from 78 +/- 8 to 93 +/- 5 microns/ml) during CPB and a significant reduction in H2O2 levels compared to Group I (P less than 0.001); plasma vitamins E and C did not change significantly in Group II.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Ascorbic Acid/BLOOD ; Cardiopulmonary Bypass/*ADVERSE EFFECTS ; Clinical Trials ; Female ; Free Radicals ; Human ; Hydrogen Peroxide/BLOOD ; Hydroxides ; Leukocytes ; Lung/CYTOLOGY ; Male ; Middle Age ; Oxygen ; Random Allocation ; Superoxide/ANTAGONISTS & INHIBITORS/*METABOLISM ; Vitamin E/BLOOD/*THERAPEUTIC USE SO - J Surg Res 1986 Jun;40(6):519-27 15 UI - 86301189 AU - Kalyanaraman B ; Hintz P ; Sealy RC TI - An electron spin resonance study of free radicals from catechol estrogens. AB - Electron spin resonance spectroscopy has been used to demonstrate production of semiquinone free radicals from the oxidation of the catechol estrogens 2- and 4-hydroxyestradiol and 2,6- and 4,6-dihydroxyestradiol. Radicals were generated by horseradish peroxidase/H2O2 or tyrosinase/O2, or by autoxidation, and were detected as their complexes with spin-stabilizing metal ions (Zn2+ and/or Mg2+). Radical production occurs via one- or two-electron oxidation of catechol estrogens, depending on the type of activating system. Autoxidation of catechol estrogens produces superoxide and H2O2 at physiological pH values. The present results also indicate a difference in the reactivity of quinones derived from 2- and 4-hydroxyestradiol. The toxicological significance of these reactions is discussed. MH - Animal ; Catechol Estrogens/*METABOLISM ; Chemistry ; Cytochrome P-450/METABOLISM ; Electron Spin Resonance ; Estradiol/ANALOGS & DERIVATIVES/METABOLISM ; Female ; Free Radicals ; Horseradish Peroxidase/METABOLISM ; Hydrogen Peroxide/PHARMACODYNAMICS ; Isoenzymes/METABOLISM ; Magnesium/PHARMACODYNAMICS ; Models, Biological ; Oxidation-Reduction ; Oxygen/PHARMACODYNAMICS ; Peroxidases/METABOLISM ; Quinones/METABOLISM/TOXICITY ; Review ; Support, U.S. Gov't, P.H.S. ; Tyrosinase/METABOLISM ; Uterus/ ENZYMOLOGY ; Zinc/PHARMACODYNAMICS SO - Fed Proc 1986 Sep;45(10):2477-84 16 UI - 86296764 AU - Mansbach CM 2d ; Rosen GM ; Rahn CA ; Strauss KE TI - Detection of free radicals as a consequence of rat intestinal cellular drug metabolism. AB - Because the intestine is the first pass organ for orally administered drugs and because some of these drugs are known to undergo oxidative metabolism leading to the formation of free radicals, we investigated the potential for this to occur in cell suspensions of rat enterocytes. As part of our study, the effect of intracellularly produced superoxide on cellular metabolism was investigated. The drugs chosen were the quinone, menadione and the aromatic nitro-containing compound, nitrazepam. On incubation of both drugs with isolated enterocytes and the spin trap, 5,5-dimethyl-1-pyrroline N-oxide (DMPO), rapid appearance of an electron paramagnetic resonance (EPR) spectrum was recorded which was characteristic of hydroxyl radicals being spin trapped by DMPO giving 2,2-dimethyl-5-hydroxy-1-pyrrolidenyloxyl (DMPO-OH). Experiments were conducted which determined that the EPR spectrum of DMPO-OH resulted from the initial spin trapping of superoxide by DMPO to yield the corresponding nitroxide, 2,2-dimethyl-5-hydroxyl-1-pyrrolidenyloxyl (DMPO-OOH). Bioreduction of DMPO-OOH by glutathione peroxidase led to the rapid formation of DMPO-OH. We believe this enzymic pathway accounted for the EPR spectrum noted in incubations with either drug in the presence of the spin trap, DMPO. The incubation of enterocytes with both drugs did not mediate release of 51Cr nor lactate dehydrogenase. However, production of 14CO2 from [14C]glucose was severely inhibited (4-5-fold) in the presence of both drugs, while the incorporation of [14C]leucine into trichloroacetic acid precipitable protein was antagonized by menadione only. We conclude that superoxide can be demonstrated to arise as the result of enterocyte metabolism of menadione or nitrazepam. The consequence of oxidative metabolism of these drugs results in cellular dysfunction. MH - Animal ; Biotransformation ; Cytochrome C/METABOLISM ; Cytochrome P-450/METABOLISM ; Electron Spin Resonance ; Free Radicals/ METABOLISM ; In Vitro ; Intestines/*METABOLISM ; Male ; Nitrazepam/METABOLISM ; Rats ; Superoxide/*METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Vitamin K/ METABOLISM SO - Biochim Biophys Acta 1986 Aug 29;888(1):1-9 17 UI - 86292350 AU - Kawabata T ; Awai M ; Kohno M TI - Generation of active oxygen species by iron nitrilotriacetate (Fe-NTA). AB - Ferric nitrilotriacetate (Fe3+-NTA) solution showed maximum absorbance at pH 7.5. The iron was in ferric high-spin state and coordinated octahedrally with a relatively symmetric structure and also probably pentagonally. A spin trapping technique employing 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) yielded a DMPO spin adduct of unknown radical with three doublets (DMPO-Z) and a simple nitroxide radical (Y-NO.) in serum from rats injected intraperitoneally with Fe3+-NTA. When the Fe3+-NTA solution was diluted 500-fold with 50 mM NTA solution, DMPO-Z, Y-NO. and an additional signal, DMPO-OH were observed. The DMPO-Z signal was suppressed by a decrease in oxygen tension, alpha-tocopherol and 3-tert-butyl-4-hydroxy-anisole (BHA). The DMPO-OH signal was suppressed in the presence of ethanol and catalase. Fe2+-NTA solution hardly produced DMPO spin adducts. The Fe3+-NTA solution produced a strong DMPO-OH signal in the presence of H2O2. Rose Bengal solution, a singlet oxygen generating system, produced the same DMPO adducts. Fe3+-NTA reacted with oxygen in solution. The oxygen was activated and might be similar to singlet molecular oxygen. In the presence of H2O2, the Fe3+-NTA solution generated a hydroxyl radical. Fe3+-NTA itself generated free radicals, but Fe2+-NTA did not. MH - Alcohol, Ethyl ; Animal ; Butylated Hydroxyanisole ; Catalase ; Cyclic N-Oxides ; Electron Spin Resonance ; Ferric Compounds/ *ANALYSIS/METABOLISM ; Free Radicals ; Iron/*ANALYSIS ; Male ; Oxygen ; Piperazines ; Rats ; Rats, Inbred Strains ; Rose Bengal ; Spectrophotometry ; Superoxide Dismutase ; Support, Non-U.S. Gov't ; Vitamin E SO - Acta Med Okayama 1986 Jun;40(3):163-73 18 UI - 86284785 AU - Van Sluys MA ; Alcantara-Gomes R ; Menck CF TI - Escherichia coli xthA mutant is not hypersensitive to ascorbic acid/copper treatment--an H2O2 generating reaction. AB - Ascorbate (vitamin C) in the presence of copper yields H2O2, which seems to be responsible for its toxic effects in bacteria. However, we found that the Escherichia coli xthA mutant strain, which is hypersensitive to H2O2, has almost the same sensitivity as the wild-type strain to ascorbate and copper treatment. Our results suggest that the DNA damage induced in E. coli by H2O2 generated in oxidized ascorbate solutions is different from that induced by direct H2O2 treatment. MH - Ascorbic Acid/*PHARMACODYNAMICS ; Bacterial Proteins/GENETICS/ PHYSIOLOGY ; Copper/*PHARMACODYNAMICS ; Drug Resistance, Microbial ; DNA Repair ; Escherichia Coli/DRUG EFFECTS/ENZYMOLOGY/ *GENETICS ; Exodeoxyribonucleases/GENETICS/PHYSIOLOGY ; Free Radicals ; Hydrogen Peroxide/*PHARMACODYNAMICS ; Oxidation-Reduction ; Support, Non-U.S. Gov't SO - Mutat Res 1986 Aug;174(4):265-9 19 UI - 86278006 AU - Wakefield LM ; Cass AE ; Radda GK TI - Electron transfer across the chromaffin granule membrane. Use of EPR to demonstrate reduction of intravesicular ascorbate radical by the extravesicular mitochondrial NADH:ascorbate radical oxidoreductase. AB - A two-compartment electron paramagnetic resonance system has been developed in which the membrane-impermeable spin probe Ni(en)2+3 is used to selectively eliminate the EPR signal from extravesicular ascorbate radical, such that radicals in intra- and extravesicular compartments can be distinguished. Using this system, we have shown that an increase in ascorbate radical in the extravesicular medium is reflected by an increase in ascorbate radical within resealed chromaffin granule ghosts containing trapped ascorbate but has no effect on radical concentrations inside liposomes containing ascorbate. This indicates that the chromaffin granule membrane contains a component, not present in liposomes, that allows equilibration between the intra- and extravesicular ascorbate/ascorbate radical couples. This component is probably cytochrome b561. We further show that activation of the mitochondrial NADH:ascorbate radical oxidoreductase in the extravesicular medium causes a decrease in intravesicular ascorbate radical in chromaffin granule ghosts but not in liposomes. These data provide direct experimental evidence for the hypothesis that the adrenal medullary mitochondrial NADH:ascorbate radical oxidoreductase could drive the re-reduction of ascorbate free radical generated inside the chromaffin granule by the turnover of dopamine beta-hydroxylase, without the ascorbate radical ever having to leave the granule. MH - Animal ; Ascorbic Acid/*METABOLISM ; Cattle ; Chromaffin Granules/ *METABOLISM ; Chromaffin System/*METABOLISM ; Cyclic N-Oxides/ PHARMACODYNAMICS ; Dopamine beta-Hydroxylase/METABOLISM ; Electron Spin Resonance ; Electron Transport ; Free Radicals ; Intracellular Membranes/*ENZYMOLOGY ; Liposomes/METABOLISM ; Mitochondria/*ENZYMOLOGY ; Nickel/METABOLISM ; NADH, NADPH Oxidoreductases/*METABOLISM ; Support, Non-U.S. Gov't SO - J Biol Chem 1986 Jul 25;261(21):9746-52 20 UI - 86278005 AU - Wakefield LM ; Cass AE ; Radda GK TI - Functional coupling between enzymes of the chromaffin granule membrane. AB - The reactions of cytochrome b561 with other redox-active components of the adrenal chromaffin granule were examined using optical difference spectroscopy. It was shown that there is no direct electron transfer between the cytochrome and dopamine beta-hydroxylase, but that in the presence of ascorbate, turnover of dopamine beta-hydroxylase causes an oxidation of the cytochrome, which is partially reversed by the action of the mitochondrial NADH:A-. oxidoreductase. Thus, these three proteins may be functionally coupled via ascorbate. A quantitative study of the relationship between the redox state of the cytochrome and the ascorbate radical concentration measured by EPR showed that ascorbate reduces the cytochrome in a one-electron transfer reaction. Generation of a proton electrochemical gradient across the granule membrane causes only a small (20 mV) increase in the cytochrome midpoint potential suggesting the cytochrome is not a proton pump. The data are consistent with a model in which cytochrome b561, by reacting with ascorbate or ascorbate free radical on either side of the granule membrane, could couple the ascorbate-consuming reaction of the dopamine beta-hydroxylase inside the chromaffin granule to the ascorbate-regenerating reaction of the NADH:A-. oxidoreductase on the outer mitochondrial membrane. The H+-ATPase of the granule membrane could both drive the flow of electrons in the direction from cytosol to granule and replenish protons consumed by the turnover of dopamine beta-hydroxylase inside the granule. MH - Animal ; Ascorbic Acid/METABOLISM ; Cattle ; Chromaffin Granules/ *ENZYMOLOGY ; Chromaffin System/*ENZYMOLOGY ; Cytochrome b/ *ANALYSIS ; Dopamine/PHARMACODYNAMICS ; Dopamine beta-Hydroxylase/ METABOLISM ; Electron Spin Resonance ; Epinephrine/ PHARMACODYNAMICS ; Ethylmaleimide/PHARMACODYNAMICS ; Free Radicals ; Fumarates/PHARMACODYNAMICS ; Fusaric Acid/ PHARMACODYNAMICS ; Intracellular Membranes/*ENZYMOLOGY ; Norepinephrine/PHARMACODYNAMICS ; NAD/METABOLISM ; Oxidation-Reduction ; Spectrophotometry ; Support, Non-U.S. Gov't ; Tyramine/PHARMACODYNAMICS SO - J Biol Chem 1986 Jul 25;261(21):9739-45 21 UI - 86274679 AU - Ekstr:om G ; Ingelman-Sundberg M TI - Mechanisms of lipid peroxidation dependent upon cytochrome P-450 LM2. AB - A mechanism of lipid peroxidation dependent on the oxidase activity of cytochrome P-450 LM2 in reconstituted membrane vesicles has been investigated. The rate of lipid peroxidation, determined as the formation of thiobarbituric-acid-reactive substances, was inhibited by CO. It increased concomitantly to the production of O-2 and H2O2, when cytochrome P-450 LM2 was incorporated into vesicles containing NADPH-cytochrome-P-450 reductase, until a 1:1 molar ratio between the enzymes was reached. Also the formation of lipid hydroperoxides was dependent on the presence of cytochrome P-450 LM2 in the membranes. This lipid peroxidation was not inhibited by hydroxyl radical scavengers and not specifically inhibited by scavengers of singlet oxygen. By contrast, superoxide dismutase was a very potent scavenger of the lipid peroxidation. A half-maximal effect at 3 ng/ml enzyme was registered, whereas a 100-fold higher concentration was necessary in order to inhibit O-2 formation as detected by succinylated cytochrome c or pyrogallol. The reason for this difference might be inherent in different types of kinetics in the interaction of O-2 with different scavengers or might possibly indicate that SOD scavenges another type of reactive oxygen, different from O-2, generated by cytochrome P-450 LM2. Iron chelators inhibited the P-450-dependent lipid peroxidation, whereas iron chelate interacted with NADPH-cytochrome-P-450 reductase in the membranes giving rise to reductase-dependent lipid peroxidation. Neither superoxide dismutase nor EDTA at high concentrations, inhibited CCl4-initiated lipid peroxidation, indicating the point of action of these compounds at the initiation step in the cytochrome-P-450-LM2-dependent lipid peroxidation. Superoxide generated by pyrogallol, in three times the amount produced by P-450 LM2, could not bring about lipid peroxidation. It is suggested that the cytochrome-P-450-dependent lipid peroxidation mechanism might be of importance for intracellular oxidative damage under certain conditions. MH - Carotene/PHARMACODYNAMICS ; Cytochrome P-450/*PHARMACODYNAMICS ; EDTA/PHARMACODYNAMICS ; Free Radicals ; Furans/PHARMACODYNAMICS ; Hydrogen Peroxide/METABOLISM ; Iron/PHARMACODYNAMICS ; Iron Chelates/PHARMACODYNAMICS ; Lipid Peroxides/*METABOLISM ; Superoxide/METABOLISM ; Superoxide Dismutase/PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Time Factors SO - Eur J Biochem 1986 Jul 1;158(1):195-201 22 UI - 86269112 AU - Hodnick WF ; Kung FS ; Roettger WJ ; Bohmont CW ; Pardini RS TI - Inhibition of mitochondrial respiration and production of toxic oxygen radicals by flavonoids. A structure-activity study. AB - A series of fourteen flavonoids were employed in a systematic structure-activity study to assess their abilities to inhibit succinoxidase and generate toxic oxygen species in beef heart mitochondria. By comparing I50 values toward succinoxidase activity, flavonoids with a catechol moiety on the b ring exhibited the following general order of potency: chalcone greater than flavone greater than flavonol greater than dihydroflavonol greater than anthocyanidin. Catechins were inactive. In a series of 3,5,7-trihydroxyflavones containing various configurations of the b ring hydroxyl groups, it was found that the flavonoids possessing adjacent trihydroxy (pyrogallol) and b ring ortho-hydroxy(catechol) configurations were the most potent inhibitors of succinoxidase, followed by those with meta-hydroxyl, monohydroxyl and unhydroxylated configurations. Four of the fifteen flavonoids tested exhibited substrate-independent, KCN-insensitive respiration. Two flavonols with a pyrogallol configuration, myricetin and quercetagetin, produced the largest respiratory bursts and were found to auto-oxidize. Evidence is presented that the mitochondrial respiratory bursts induced by both flavonols and their auto-oxidation resulted in the generation of O-2 and H2O2. MH - Animal ; Cattle ; Chalcone/*ANALOGS & DERIVATIVES/ PHARMACODYNAMICS ; Flavones/*PHARMACODYNAMICS ; Free Radicals ; Mitochondria, Heart/*DRUG EFFECTS ; Oxidation-Reduction ; Oxidoreductases/ANTAGONISTS & INHIBITORS ; Oxygen/*METABOLISM ; Oxygen Consumption/DRUG EFFECTS ; *Propiophenones ; Spectrum Analysis ; Structure-Activity Relationship SO - Biochem Pharmacol 1986 Jul 15;35(14):2345-57 23 UI - 86253397 AU - Bray TM ; Kubow S ; Bettger WJ TI - Effect of dietary zinc on endogenous free radical production in rat lung microsomes. AB - The objective of this study was to investigate the effect of dietary zinc on endogenous production of free radicals in lung and liver microsomes. Male weanling rats were fed a zinc-deficient basal diet containing less than 1.1 ppm zinc, or were pair-fed or fed ad libitum a zinc-adequate diet supplemented with 100 ppm zinc. The isolated microsomes (100,000 X g precipitate) of lung and liver were incubated with 0.1 M PBN (spin trap) and 0.3 mM NADPH (cofactor) at 37 degrees C for 1.0 h. A carbon-centered free radical (aN = 16.0 G, aH beta = 3.4 G) was trapped in both lung and liver microsomes. There was a significant increase in the concentration of carbon-centered free radicals generated in lung microsomes in animals fed a zinc-deficient diet. Dietary zinc status did not significantly affect the concentration of free radicals in liver microsomes. The amount of free radicals generated is proportional to microsomal protein concentration and is linear with protein concentration between 5 and 20 mg per milliliter of incubate. The free radicals formed in the microsomal system were dependent on the presence of NADPH. Carbon monoxide inhibited 40-50% of the free radical production in both lung and liver microsomes. The results suggest that dietary zinc deficiency stimulates the production of endogenous free radicals in rat lung microsomes by an NADPH- and cytochrome P-450-dependent system. MH - Animal ; Body Weight ; Carbon Monoxide/PHARMACODYNAMICS ; Diet ; Electron Spin Resonance ; Free Radicals ; Lung/DRUG EFFECTS/ *METABOLISM ; Male ; Microsomes/METABOLISM ; Microsomes, Liver/ DRUG EFFECTS/METABOLISM ; NADP/PHARMACODYNAMICS ; Rats ; Rats, Inbred Strains ; Sulfates/PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Zinc/ADMINISTRATION & DOSAGE/*DEFICIENCY/PHARMACODYNAMICS SO - J Nutr 1986 Jun;116(6):1054-60 24 UI - 86245454 AU - Gabrielson EW ; Rosen GM ; Grafstrom RC ; Strauss KE ; Harris CC TI - Studies on the role of oxygen radicals in asbestos-induced cytopathology of cultured human lung mesothelial cells. AB - The possible role of oxygen radicals in mediating the cytopathologic effects of asbestos was studied using human mesothelial cells in culture. Electron paramagnetic resonance measurements of intact cells using the spin trap 5,5-dimethyl-1-pyrroline-1-oxide failed to detect any increase in oxygen radicals in mesothelial cells after exposure to amosite asbestos, although oxygen radicals were readily detected in cells exposed to menadione, an uncoupler of oxidation-reduction reactions. Cellular thiol levels were reduced after exposure to menadione, but were not affected by exposure to asbestos. Addition to the culture media of the free radical scavengers superoxide dismutase, reduced glutathione, N-acetylcysteine, or D-alpha-tocopherol had no affect on the dose-dependent cytotoxicity of amosite fibers. Furthermore, exposure of the mesothelial cells to amosite fibers resulted in no significant increase in the level of DNA single-strand breaks. These results all suggest that for cultured human mesothelial cells, oxygen free radicals are not important mediators of the cytopathic effect of asbestos. MH - Asbestos/*TOXICITY ; Cell Count ; Cells, Cultured ; DNA/ANALYSIS ; Electron Spin Resonance ; Free Radicals ; Human ; In Vitro ; Lung/CYTOLOGY/*DRUG EFFECTS ; *Oxygen ; Spin Labels ; Sulfhydryl Compounds/ANALYSIS ; Support, Non-U.S. Gov't ; Vitamin K/TOXICITY SO - Carcinogenesis 1986 Jul;7(7):1161-4 25 UI - 86242174 AU - Wojtczak L ; Szewczyk A TI - Internalization of the spin-labelled surface potential probe CAT12 by energized mitochondria. AB - Rat liver mitochondria briefly incubated in the presence of 50 microM 4-(dodecyl dimethyl ammonium)-1-oxyl-2,2,6,6-tetramethyl piperidine bromide (CAT12) bind 1.9 nmol of this spin-labelled membrane probe per mg mitochondrial protein in a form which is not reducible by ascorbate. Upon energization with ATP the amount of the non-reducible CAT12 increases by about 37%. It is concluded that CAT12 which is not reducible by externally added ascorbate is bound to the inner surface of the inner mitochondrial membrane and/or accumulated in the matrix compartment. Therefore, CAT12 is not suitable to monitor the surface potential of mitochondria and other organelles which develop a transmembrane potential, negative inside. MH - Adenosine Triphosphate/METABOLISM ; Animal ; Ascorbic Acid/ PHARMACODYNAMICS ; Carbonyl Cyanide m-Chlorophenyl Hydrazone/ PHARMACODYNAMICS ; Cyclic N-Oxides/*METABOLISM ; Intracellular Membranes/METABOLISM ; Mitochondria, Liver/*METABOLISM ; Rats ; Spin Labels/*METABOLISM ; Time Factors SO - Biochem Biophys Res Commun 1986 May 14;136(3):941-6 26 UI - 86231113 AU - Jendryczko A ; Dro:zd:z M ; Tomala J ; Magner K TI - Copper and zinc concentrations, and superoxide dismutase activities in malignant and nonmalignant tissues of female reproductive organs. AB - The copper and zinc concentrations in 44 malignant and 48 nonmalignant women tissue samples of reproductive organs in women were measured. In malignant samples, the mean copper concentrations were 110%, 76%, and 38% higher for cervix, endometrium and ovary than the nonmalignant ones. The zinc concentrations in the analysed malignant tissues were lower than that in the corresponding nonmalignant tissues. The results of superoxide dismutase activities determinations demonstrate a considerable lowering of the enzymatic capacities to remove free oxygen radical in malignant tissues. A hypothesis for possible mechanism involving elevated copper concentrations, and decreased zinc concentrations, which may be responsible for malignant processes, are presented. MH - Copper/*ANALYSIS ; Female ; Free Radicals ; Genital Neoplasms, Female/*ANALYSIS ; Genitalia, Female/*ANALYSIS ; Human ; Superoxide Dismutase/*ANALYSIS ; Zinc/*ANALYSIS SO - Neoplasma 1986;33(2):239-44 27 UI - 86223969 AU - Harwood HJ Jr ; Greene YJ ; Stacpoole PW TI - Inhibition of human leukocyte 3-hydroxy-3-methylglutaryl coenzyme A reductase activity by ascorbic acid. An effect mediated by the free radical monodehydroascorbate. AB - 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity in microsomes isolated from cultured lymphoid (IM-9) cells or freshly isolated human leukocytes was markedly decreased by either ascorbic acid or its oxidized derivative, dehydroascorbate. Inhibition of IM-9 leukocyte HMG-CoA reductase activity was log linear between 0.01 and 10 mM ascorbic acid (25 and 81% inhibition, respectively) and 0.1 and 10 mM dehydroascorbate (5 and 75% inhibition, respectively). Inhibition was noncompetitive with respect to HMG-CoA (Km = 10.2 microM (RS); ascorbic acid, Ki = 6.4 mM; dehydroascorbate, Ki = 15 mM) and competitive with respect to NADPH (Km = 16.3 microM; acetic acid, Ki = 6.3 mM; dehydroascorbate, Ki = 3.1 mM). Ascorbic acid and dehydroascorbate are interconverted through the free radical intermediate monodehydroascorbate. Reducing agents are required to convert dehydroascorbate to monodehydroascorbate, but prevent formation of the free radical from ascorbate. In microsomes from IM-9 cells, the reducing agent, dithiothreitol, abolished HMG-CoA reductase inhibition by ascorbate but enhanced inhibition by dehydroascorbate. In addition, the concentration of monodehydroascorbate present in ascorbate solutions was directly proportional to the degree of HMG-CoA reductase inhibition by 1.0 mM ascorbate. Fifty per cent inhibition of enzyme activity occurred at a monodehydroascorbate concentration of 14 microM. These data indicate that monodehydroascorbate mediates inhibition of HMG-CoA reductase by both ascorbate and dehydroascorbate. This effect does not appear to be due to free radical-induced membrane lipid modification, however, since both ascorbate and dehydroascorbate inhibited the protease-solubilized, partially purified human liver enzyme. Since inhibition of HMG-CoA reductase occurs at physiological concentrations of ascorbic acid in the human leukocyte (0.2-1.72 mM), this vitamin may be important in the regulation of endogenous cholesterol synthesis in man. MH - Ascorbic Acid/*ANALOGS & DERIVATIVES/METABOLISM/*PHARMACODYNAMICS ; Dehydroascorbic Acid/*ANALOGS & DERIVATIVES/METABOLISM/ PHARMACODYNAMICS ; Dithiothreitol/PHARMACODYNAMICS ; Free Radicals ; Glutathione/PHYSIOLOGY ; Human ; Hydroxymethylglutaryl CoA Reductases/*ANTAGONISTS & INHIBITORS/BLOOD ; Kinetics ; Leukocytes/*ENZYMOLOGY ; NADP/PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - J Biol Chem 1986 Jun 5;261(16):7127-35 28 UI - 86215320 AU - Davison AJ ; Legault NA ; Steele DW TI - Effect of 6-hydroxydopamine on polymerization of tubulin. Protection by superoxide dismutase, catalase, or anaerobic conditions. AB - Microtubular protein (tubulin) isolated from porcine brain was subjected to selected oxidative stresses, including incubation with the neurotoxin 6-hydroxydopamine (6-OHDA) under aerobic and anaerobic conditions. The functional capacity of the tubulin was determined on the basis of its ability to form microtubules as measured by alterations in the viscosity of the test mixtures, and confirmed by electron microscopy. 6-OHDA completely inhibited formation of microtubules at concentrations as low as 10 mM. Assembled microtubules were half as susceptible to destruction by 6-OHDA as unaggregated tubulin. Anaerobic conditions or the presence of catalase, superoxide dismutase, or a mixture of superoxide dismutase and catalase provided partial protection against 6-OHDA-induced destruction. In control reactions, tubulin-containing solutions incubated for up to 8 hr at ambient oxygen tensions, also showed significant decreases in ability to polymerize. Anaerobic conditions provided partial protection against this loss of function. In contrast, ascorbate accelerated the loss of activity upon standing, while glutathione or dithiothreitol offered no protection. MH - Anaerobiosis ; Animal ; Ascorbic Acid/PHARMACODYNAMICS ; Buffers ; Catalase/*PHARMACODYNAMICS ; Dithiothreitol/PHARMACODYNAMICS ; Free Radicals ; Glutathione/PHARMACODYNAMICS ; Hydroxydopamines/ *TOXICITY ; Microscopy, Electron ; Microtubules/DRUG EFFECTS ; Polymers/*METABOLISM ; Quinones/PHARMACODYNAMICS ; Superoxide Dismutase/*PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Swine ; Tubulin/*METABOLISM SO - Biochem Pharmacol 1986 May 1;35(9):1411-7 29 UI - 86215306 AU - Gorus FK ; Finsy R ; Pipeleers DG TI - Alloxan toxicity in human and canine spermatozoa. Possible biochemical basis for a species difference in sensitivity. AB - In view of the well known species differences in the sensitivity of pancreatic B-cells to the toxic glucose analogue alloxan, it was tested whether spermatozoa from two species with a different diabetogenic effect of alloxan displayed a similar difference in their sensitivity to this drug. In canine spermatozoa, less than 2 mM alloxan profoundly reduced the rate of glucose oxidation and cellular motility whereas more than 5 mM was required to significantly alter these parameters in human spermatozoa. Such species difference was not observed in spermatozoal sensitivity towards the inhibitory effects of tert-butyl hydroperoxide. The phenomenon is not attributable to a different rate of alloxan uptake since the drug is not incorporated by dog or human spermatozoa. The alloxan toxicity was counteracted by D-glucose and its 3-O-methyl analogue in both species, and was potentiated by ascorbic acid; however, only in man. The protective effect of D-glucose was much less marked in tert-butyl hydroperoxide-cytotoxicity. It is concluded that the observed species difference in spermatozoal alloxan sensitivity is not related to differences in alloxan uptake or in sensitivity to organic peroxides; differences in cellular scavenging of superoxide anion radicals and/or ascorbic acid metabolism may explain the lower sensitivity of human spermatozoa for alloxan. MH - Adult ; Alloxan/METABOLISM/*TOXICITY ; Animal ; Ascorbic Acid/ PHARMACODYNAMICS ; Dogs ; Free Radicals ; Glucose/ PHARMACODYNAMICS ; Human ; Male ; Species Specificity ; Sperm Motility/DRUG EFFECTS ; Spermatozoa/*DRUG EFFECTS/METABOLISM ; Superoxide/METABOLISM ; Support, Non-U.S. Gov't SO - Biochem Pharmacol 1986 May 15;35(10):1725-9 30 UI - 86215156 AU - Wustmann C ; Fischer HD ; Schmidt J TI - Protective and restitutive effects of antihypoxic drugs on posthypoxic dopamine release inhibition. AB - Exposure of rats to hypoxia results in a substantial decrease of dopamine release from striatum slices for several days. Nootropic drugs (piracetam, meclofenoxate hydrochloride, methylglucamine orotate, nicergoline) accelerate the restitution of posthypoxic release inhibition. In contrast, amphetamine is ineffective in this respect. The antihypoxic action of sedatives (diazepam, phenobarbital) prevents the decrease of dopamine release. Comparable results with free radical scavengers (cysteamine hydrochloride, sodium formiate, ouabain), ascorbic acid, natrii calcii edetas, selenium methionine and acetylsalicylic acid which protect dopamine release from hypoxically produced changes agree with and support the hypothesis of hypoxia induced free radical generation followed by phospholipid peroxidation altering particularly neuronal membrane function. On that account, dopamine release from rat striatum slices reflects not only the vulnerability of neuronal membrane function by hypoxia but also the preventive, protective and restitutive effects of antihypoxic drugs of different type and is able to contribute to discriminating drug investigation. MH - Amphetamine/PHARMACODYNAMICS ; Animal ; Anoxia/DRUG THERAPY/ *METABOLISM ; Antioxidants/PHARMACODYNAMICS ; Chelating Agents/ PHARMACODYNAMICS ; Corpus Striatum/METABOLISM ; Dopamine/ *METABOLISM ; Free Radicals ; Lipid Peroxides/METABOLISM ; Male ; Piracetam/*PHARMACODYNAMICS ; Pyrrolidinones/*PHARMACODYNAMICS ; Rats ; Selenium/PHARMACODYNAMICS SO - Biomed Biochim Acta 1986;45(4):549-56 31 UI - 86201214 AU - Chester JF ; Gaissert HA ; Ross JS ; Malt RA ; Weitzman SA TI - Augmentation of 1,2-dimethylhydrazine-induced colon cancer by experimental colitis in mice: role of dietary vitamin E. AB - Because ulcerative colitis predisposes to colonic cancer, for determination of the effect of colitis on experimental colon carcinogenesis, rectal instillations of peptides that attract and activate neutrophils were used to induce colitis in CD-1 (ICR) BR mice receiving 20 weekly injections of the carcinogen 1,2-dimethylhydrazine [(DMH) CAS: 540-73-8]. From week 4 through week 15 of DMH injections, twice-weekly enemas of formyl-norleucyl-leucyl-phenylalanine were given to DMH-treated mice. The effect of the antioxidant vitamin E in the diet (1,750 IU/kg diet) was studied in another group of mice treated with DMH and having colitis. Four weeks after DMH was discontinued, cancer occurred in 9 of 28 (32%) animals with DMH plus control enemas, in 22 of 29 (76%) animals with DMH plus colitis (P = .001), and in 16 of 28 (57%) animals with DMH plus colitis plus supplemental vitamin E (P = .11 compared with the group with DMH and colitis). Colitis enhances DMH-induced colonic carcinogenesis. MH - Animal ; Cell Division ; Colitis/*COMPLICATIONS ; Colonic Neoplasms/CHEMICALLY INDUCED/*ETIOLOGY/PREVENTION & CONTROL ; Diet ; Dimethylhydrazines ; Free Radicals ; Male ; Mice ; Mice, Inbred Strains ; Support, Non-U.S. Gov't ; Vitamin E/ *PHARMACODYNAMICS SO - JNCI 1986 May;76(5):939-42 32 UI - 86201137 AU - Sugihara K ; Gemba M TI - Modification of cisplatin toxicity by antioxidants. AB - cis-Diamminedichloroplatinum II (cisplatin) is a potent anticancer chemotherapeutic agent. The major limitation in its use is nephrotoxicity, caused by an unknown mechanism. Injection of cisplatin into rats caused a decrease in body weight and an increase in blood urea nitrogen (BUN). These effects were modified by giving a radical scavenger, alpha-tocopherol, before the cisplatin injection. N-N'-diphenyl-p-phenylenediamine, another powerful radical scavenger, also attenuated the increase in BUN induced by cisplatin. These results suggest that the toxic effects of cisplatin may be related to free radical induced damage. MH - Animal ; Antioxidants/*PHARMACODYNAMICS ; Blood Urea Nitrogen ; Body Weight/DRUG EFFECTS ; Cisplatin/*TOXICITY ; Free Radicals ; Kidney/DRUG EFFECTS ; Lipid Peroxides/METABOLISM ; Male ; Phenylenediamines/PHARMACODYNAMICS ; Rats ; Rats, Inbred Strains ; Vitamin E/PHARMACODYNAMICS SO - Jpn J Pharmacol 1986 Feb;40(2):353-5 33 UI - 86191395 AU - Riley MV ; Schwartz CA ; Peters MI TI - Interactions of ascorbate and H2O2: implications for in vitro studies of lens and cornea. AB - The interaction of ascorbate, hydrogen peroxide and oxygen has been examined in order to understand the equilibrium between these compounds that exists in the aqueous humor of the eye and their influence on function of the corneal endothelium. Ascorbate was found to promote corneal swelling when isolated corneas were perfused with a medium lacking glucose. This was found to be due to the rapid oxidation of ascorbate in the medium, yielding H2O2 which is toxic to the endothelial cells. In the absence of oxygen, or if EDTA was added to the medium, no H2O2 was produced from ascorbate, but ascorbate reacted with any pre-existing H2O2. Oxidation of ascorbate in the aqueous humor is limited by the presence of glutathione (and, possibly, other compounds) and no significant increase in H2O2 concentration occurs on standing in air. Nevertheless, the concentration of H2O2 in the aqueous is directly dependent on the concentration of ascorbate secreted in the aqueous humor. Therefore, there must be a dynamic equilibrium in this fluid between ascorbate, H2O2 and oxygen, and it may be modulated by glutathione. Each of these substances is important in redox reactions, including free-radical production or scavenging. Consequently, when studying the effects on corneal or lenticular function of other agents which cause or relieve oxidant stress, it is critical that the modifying effects of ascorbate and H2O2, as they occur in vivo, be considered. A perfusion system is described which permits an approximation in vitro of stable concentrations of ascorbate, H2O2, GSH and O2 similar to those found in the aqueous humor. MH - Animal ; Aqueous Humor/METABOLISM ; Ascorbic Acid/*METABOLISM/ PHARMACODYNAMICS ; Cornea/DRUG EFFECTS/*METABOLISM ; Free Radicals ; Hydrogen Peroxide/*METABOLISM ; In Vitro ; Lens, Crystalline/*METABOLISM ; Oxidation-Reduction ; Oxygen/METABOLISM ; Perfusion ; Rabbits ; Support, U.S. Gov't, P.H.S. SO - Curr Eye Res 1986 Mar;5(3):207-16 34 UI - 86184445 AU - Malmgren R ; Unge G ; Zetterstr:om O ; Theorell H ; de Wahl K TI - Lowered glutathione-peroxidase activity in asthmatic patients with food and aspirin intolerance. AB - In analogy with findings from animal experiments, people with low glutathione-peroxidase (GSH-Px) activity could be expected to have altered sensitivities to effects of drugs, chemicals and possibly food. We have investigated GSH-Px activity in 12 patients with intrinsic asthma and food and aspirin intolerance. Ten of the 12 patients had very low or low GSH-Px activity and the frequency of low GSH-Px activity in this group was statistically significant (P less than 0.001) compared with the control material of age- and sex-matched healthy individuals. Our finding of lowered GSH-Px activity in patients with aspirin intolerance may indicate the involvement of hitherto unknown mechanisms in the pathogenesis of asthmatic disorders. MH - Arachidonic Acids/METABOLISM ; Aspirin/ADVERSE EFFECTS/ PHARMACODYNAMICS ; Asthma/BLOOD/COMPLICATIONS/*ENZYMOLOGY ; Blood Platelets/ENZYMOLOGY/PHYSIOLOGY ; Drug Hypersensitivity/ *COMPLICATIONS/ENZYMOLOGY ; Food Hypersensitivity/*COMPLICATIONS/ ENZYMOLOGY ; Free Radicals ; Glutathione Peroxidase/*BLOOD/ PHYSIOLOGY ; Prostaglandin Synthase/ANTAGONISTS & INHIBITORS/ METABOLISM ; Selenium/DEFICIENCY/PHYSIOLOGY ; Support, Non-U.S. Gov't SO - Allergy 1986 Jan;41(1):43-5 35 UI - 86155709 AU - Del Principe D ; Menichelli A ; Lubrano R ; Bandino D ; Di Giulio S ; Di Corpo ML ; Giardini O TI - Vitamin E consumption by human blood platelets activated by latex particles. AB - Human blood platelet activation elicited by latex particles is associated to a 30% decrease in the cellular content of vitamin E. The vitamin E consumption is inhibited by the addition of catalase (500 U/ml) and azide (1 mM), but it is not affected by potassium cyanide (1 mM). It may be proposed that the challenge of platelets with particulate stimuli causes generation of oxygen reduction products, which leads to vitamin E depletion. MH - Azides/PHARMACODYNAMICS ; Blood Platelets/DRUG EFFECTS/ *METABOLISM/PHYSIOLOGY ; Catalase/PHARMACODYNAMICS ; Free Radicals ; Human ; Latex ; Malondialdehyde/BIOSYNTHESIS ; Microspheres ; Oxygen/METABOLISM ; Support, Non-U.S. Gov't ; Thromboxane B2/METABOLISM ; Vitamin E/*METABOLISM SO - Am J Hematol 1986 Apr;21(4):351-6 36 UI - 86154945 AU - Hermansen K ; Wassermann K TI - The effect of vitamin E and selenium on doxorubicin (Adriamycin) induced delayed toxicity in mice. AB - The antagonistic action of repeated administration of vitamin E and selenium on the lethality caused by a single intraperitoneal injection of doxorubicin 15 to 20 mg/kg has been investigated in mice. Mice were treated with vitamin E, 20 to 4100 mg/kg intraperitoneally/intramuscularly daily or once a week and/or selenium 27 micrograms/kg orally daily for 6 to 7 weeks, i.e. one or two weeks before and five weeks after doxorubicin administration. 30 mg/kg of doxorubicin intraperitoneally caused 100% lethality within 4 days, whereas 15 mg/kg caused no deaths within a week, but resulted in a delayed toxicity with a cumulative lethality of 80% at the end of the observation period of 6 months. Vitamin E protected the mice from the lethal effect of doxorubicin, 15 mg/kg during the administration, but the mice began to die when the vitamin E administration was discontinued. Selenium only protected the mice for two weeks in spite of the continuous administration of selenium. The combined administration of vitamin E and selenium had no protective effect on the lethality caused by doxorubicin, 20 mg/kg. The pathogenesis of the delayed lethality of a single doxorubicin administration in mice is discussed. It is concluded that vitamin E and/or selenium have no significant protective action against doxorubicin induced delayed lethality in mice. MH - Administration, Oral ; Animal ; Doxorubicin/*TOXICITY ; Female ; Free Radicals ; Heart/DRUG EFFECTS ; Injections, Intramuscular ; Injections, Intraperitoneal ; Mice ; Mice, Inbred BALB C ; Myocardium/METABOLISM ; Selenium/*PHARMACODYNAMICS ; Vitamin E/ *PHARMACODYNAMICS SO - Acta Pharmacol Toxicol (Copenh) 1986 Jan;58(1):31-7 37 UI - 86149588 AU - Maridonneau-Parini I ; Braquet P ; Garay RP TI - Heterogeneous effect of flavonoids on K+ loss and lipid peroxidation induced by oxygen-free radicals in human red cells. AB - Treatment of fresh erythrocytes with phenazine methosulfate, an intracellular generator of oxygen-free radicals, and diethyldithiocarbamate an inhibitor of superoxide dismutase results in membrane damage consisting in lipid peroxidation and increase in passive K+ permeability. Various flavonoids which have previously been reported to act as oxygen-free radical scavengers were tested on this erythrocyte model. Surprisingly, flavonoids did not exhibit the same effect on the oxygen free radical-stimulated K+ permeability. It was possible to classify these agents into four groups: protective (those decreasing the oxygen-free radical-stimulated K+ permeability): kaempferol, naringenin, apigenin, naringin; toxic (those increasing the deleterious effect of oxygen-free radicals): myricetin, delphinidin, quercetin; biphasic effective (characterized by opposite effects depending on the concentration): phloretin, cyanin, catechin, morin and inactive: rutin, phloridzin. In addition, a similar classification was observed when membrane lipid peroxidation was examined, i.e. kaempferol decreased lipid peroxide formation whereas myricetin enhanced it, morin exhibited a biphasic effect and rutin has no effect. The previously reported metal chelating effect of flavonoids could not totally explain the protective effect of kaempferol as was demonstrated by the partial protective effect exhibited by desferrioxamine. Moreover, this study suggests that a generation of oxygen-free radicals in red cells induced a K+ loss which probably results from membrane lipid peroxidation. MH - Bioflavonoids/*PHARMACODYNAMICS ; Cell Membrane Permeability ; Deferoxamine/PHARMACODYNAMICS ; Diethyldithiocarbamate/ PHARMACODYNAMICS ; Erythrocytes/DRUG EFFECTS/*METABOLISM ; Free Radicals ; Human ; In Vitro ; Lipid Peroxides/*BLOOD ; Malondialdehyde/BLOOD ; Potassium/*BLOOD ; Superoxide Dismutase/ BLOOD SO - Pharmacol Res Commun 1986 Jan;18(1):61-72 38 UI - 86140134 AU - Tan SL ; Kopczynski MG ; Bachovchin WW ; Orme-Johnson WH ; Babior BM TI - Electron spin-echo studies of the composition of the paramagnetic intermediate formed during the deamination of propanolamine by ethanolamine ammonia-lyase, and AdoCbl-dependent enzyme. AB - During the deamination of S-2-aminopropanol by the AdoCbl-dependent ethanolamine ammonia-lyase of Clostridia sp., a catalytic intermediate accumulates whose active site contains two paramagnetic species: cob(II)alamin and a free radical derived from the substrate molecule. Spin-echo spectroscopy has revealed that the unpaired electron on the substrate-derived radical is delocalized over a nitrogen atom that from its quadrupole splittings is probably a component of a secondary amide group. Experiments with 15N- and deuterium-labeled propanolamine gave no evidence of an interaction between this unpaired electron and the nitrogen originally attached to the substrate molecule. These results strongly suggest that the substrate-derived radical in this intermediate has already lost its nitrogen, and that this radical is stabilized by delocalization of the unpaired electron onto a nitrogen most likely situated in one of the peptide bonds of the enzyme backbone. MH - Ammonia-Lyases/*PHARMACODYNAMICS ; Cobamides/*PHARMACODYNAMICS ; Deamination ; Ethanolamine Ammonia-Lyase/*PHARMACODYNAMICS ; Free Radicals ; Nitrogen ; *Propanolamines ; Spectrum Analysis ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. SO - J Biol Chem 1986 Mar 15;261(8):3483-5 39 UI - 86138929 AU - J:avor T ; T:arnok F ; Past T ; Nagy S TI - Cytoprotective effect of free radical scavengers against mucosal damage produced by different antirheumatic drugs. AB - Following 200 mg aspirin, 20 mg indomethacin or 100 mg diclofenac, gastric mucosal damage was evoked after five hours in rats. By administering vitamin A, vitamin E, MTDQ (6,6-methylenebis-2,2,4-trimethyl-1,2-dihydroquinoline), vitamin C, lipoic acid and penicillamine intragastrically at the time of the application of the damaging agent, the authors studied the beneficial effect of these free-radical scavengers upon the mucosal lesions. Vitamin C and penicillamine exerted no significant protective effect. Among the other drugs, the most effective were the lipid-soluble ones: vitamin A, vitamin E and MTDQ. The authors hypothesized that the gastric damage may be connected with the degradation of the polyunsaturated fatty acid components of the cellular membranes and thus the lipid-soluble free radical scavengers were able to offer protection. MH - Animal ; Anti-Inflammatory Agents/*ANTAGONISTS & INHIBITORS ; Anti-Ulcer Agents ; Antioxidants/*PHARMACODYNAMICS ; Ascorbic Acid/PHARMACODYNAMICS ; Aspirin/ANTAGONISTS & INHIBITORS ; Diclofenac/ANTAGONISTS & INHIBITORS ; Female ; *Free Radicals ; Gastric Mucosa/*DRUG EFFECTS ; Indomethacin/ANTAGONISTS & INHIBITORS ; Lipoic Acid/PHARMACODYNAMICS ; Male ; Penicillamine/ PHARMACODYNAMICS ; Quinolines/PHARMACODYNAMICS ; Rats ; Solubility ; Vitamin A/PHARMACODYNAMICS ; Vitamin E/ PHARMACODYNAMICS SO - Int J Tissue React 1986;8(1):35-40 40 UI - 86136440 AU - Valenzuela A ; Guerra R TI - Differential effect of silybin on the Fe2+-ADP and t-butyl hydroperoxide-induced microsomal lipid peroxidation. AB - We have observed a differential effect of silybin dihemisuccinate on rat liver microsomal oxygen consumption and on lipid peroxidation induced by NADPH-Fe2+-ADP and t-butyl hydroperoxide. These results are ascribed to the antioxidant properties of the flavonoid. The differences observed in the effect of the catalysts may be a consequence of the different capacity of silybin to act as a scavenger of free radicals formed by NADPH-Fe2+-ADP or t-butyl hydroperoxide. MH - Adenosine Diphosphate/*PHARMACODYNAMICS ; Animal ; Ferrous Compounds/*PHARMACODYNAMICS ; Flavones/*PHARMACODYNAMICS ; Free Radicals ; Iron/*PHARMACODYNAMICS ; Lipid Peroxides/*METABOLISM ; Male ; Microsomes, Liver/DRUG EFFECTS/*METABOLISM ; NADP/ PHARMACODYNAMICS ; Oxygen Consumption/DRUG EFFECTS ; Peroxides/ *PHARMACODYNAMICS ; Rats ; Rats, Inbred Strains ; Silymarin/ ANALOGS & DERIVATIVES/*PHARMACODYNAMICS ; Support, Non-U.S. Gov't SO - Experientia 1986 Feb 15;42(2):139-41 41 UI - 86123466 AU - Marubayashi S ; Dohi K ; Ochi K ; Kawasaki T TI - Role of free radicals in ischemic rat liver cell injury: prevention of damage by alpha-tocopherol administration. AB - The present study was undertaken to determine whether alpha-tocopherol pretreatment could modify cellular free radical metabolism during hepatic ischemia and subsequent reperfusion and prolong the viability of the liver. Although ischemia of the liver for 90 minutes did not permit survival of the animals, alpha-tocopherol administration (10 mg/kg of body weight) for 3 days increased the survival rate to 45.5%. The period of ischemia was accompanied by decreases in the hepatic adenosine triphosphate (ATP) level, endogenous alpha-tocopherol, and total glutathione (reduced and oxidized) without any significant increase in endogenous coenzyme Q (CoQ) homologs (CoQ9 and CoQ10) and lipid peroxide formation. The subsequent restoration of blood flow resulted in a low recovery of ATP and marked decreases in endogenous alpha-tocopherol, total glutathione, and CoQ homologs and, on the contrary, a marked increase in lipid peroxide levels. In alpha-tocopherol-treated animals, however, resynthesis of ATP was accelerated even after 90 minutes of ischemia, and there were no changes in the levels of total glutathione or CoQ homologs or in the level of the enhanced alpha-tocopherol during the reperfusion period. The pretreatment also completely suppressed the elevation of lipid peroxide levels. These results are compatible with the assumption that cellular damage caused by hepatic ischemia can be explained by free radical reaction processes during ischemia and especially reperfusion and suggest that administration of a free radical scavenger and antioxidant, alpha-tocopherol, is effective in ischemic liver cell injury. MH - Adenine Nucleotides/METABOLISM ; Animal ; *Free Radicals ; Glutathione/ANALOGS & DERIVATIVES/METABOLISM ; Glutathione Peroxidase/METABOLISM ; Ischemia/*METABOLISM ; Lipid Peroxides/ BIOSYNTHESIS ; Liver/*BLOOD SUPPLY/DRUG EFFECTS/METABOLISM ; Male ; Mitochondria, Liver/DRUG EFFECTS ; Rats ; Rats, Inbred Strains ; Ubiquinone/METABOLISM ; Vitamin E/*PHARMACODYNAMICS SO - Surgery 1986 Feb;99(2):184-92 42 UI - 86116722 AU - Mossman BT ; Marsh JP ; Shatos MA TI - Alteration of superoxide dismutase activity in tracheal epithelial cells by asbestos and inhibition of cytotoxicity by antioxidants. AB - We report here the inhibition of asbestos-induced cytotoxicity in a hamster tracheal epithelial cell line by superoxide dismutase, a scavenger of superoxide (O2-.), and by mannitol and dimethylthiourea, scavengers of the hydroxyl radical (OH.). By using these agents, cell damage was ameliorated in cultures exposed to long (greater than 10 microns in length) fibers of chrysotile and crocidolite asbestos. In contrast, injury to epithelial cells by short (less than or equal to 2 microns) chrysotile or glass fibers was not prevented by scavengers of O2-., OH., H2O2 or 1O2 (singlet oxygen). These results implicate active oxygen species as mediators of injury by long asbestos fibers to cells of the respiratory tract. By using immunocytochemical and biochemical techniques, we detected appreciable amounts of copper-zinc superoxide dismutase in hamster tracheobronchial epithelial cells and alveolar macrophages in vitro and in histologic sections of rat and human respiratory tract. Activity of total endogenous superoxide dismutase (copper-zinc and manganese forms) increased in tracheal epithelial cells exposed for several days in vitro to either crocidolite or chrysotile asbestos but was unchanged in untreated cells and those exposed to comparable amounts of glass fibers. After inhalation of asbestos by rats, or exposure of cells in culture to asbestos, long fibers were observed protruding from both epithelial cells and alveolar macrophages. The unsuccessful phagocytosis of long fibers of asbestos coupled with generation of oxygen free radicals might explain the increased pathogenic potential of long fibers in asbestos-associated diseases of the respiratory tract. MH - Animal ; Antioxidants/*PHARMACODYNAMICS ; Asbestos/ANTAGONISTS & INHIBITORS/*PHARMACODYNAMICS ; Clone Cells ; Epithelium/CYTOLOGY/ DRUG EFFECTS/ENZYMOLOGY ; Free Radicals ; Hamsters ; Histocytochemistry ; Immunochemistry ; Microscopy, Electron ; Minerals/PHARMACODYNAMICS ; Oxygen/METABOLISM ; Radioisotopes/ DIAGNOSTIC USE ; Selenium/DIAGNOSTIC USE/METABOLISM ; Superoxide Dismutase/*METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Trachea/CYTOLOGY/DRUG EFFECTS/*ENZYMOLOGY SO - Lab Invest 1986 Feb;54(2):204-12 43 UI - 86111750 AU - Mak IT ; Kramer JH ; Weglicki WB TI - Potentiation of free radical-induced lipid peroxidative injury to sarcolemmal membranes by lipid amphiphiles. AB - The effects of naturally occurring lipid amphiphiles on free radical-mediated peroxidative injury in isolated canine sarcolemma were studied. Highly enriched canine myocytic sarcolemmal membranes were preincubated for 10 min at 37 degrees C with or without different amphiphilic lipids before the addition of a free radical-generating system consisting of dihydroxyfumarate and Fe3+-ADP. Lipid peroxidation, assayed as malondialdehyde formation, was catalyzed linearly up to 40 min in the control samples. Pretreatment of the sarcolemma with palmitoyl-CoA, palmitoylcarnitine, or lysophosphatidylcholine accelerated the initial rates (20 min) of peroxidation in a concentration-dependent manner (10-100 microM) and achieved maximal stimulation (240%, 160%, and 210%, respectively, of controls) at 50 microM concentrations of each of these amphiphiles. However, free fatty acids, CoA, and carnitine were without effect. These promoting effects of the amphiphiles persisted over a wide pH range (pH 6.0-7.8) and exhibited additive effects when lower levels of different amphiphiles were combined together. Associated with the accelerated rates of peroxidation produced by palmitoyl-CoA and palmitoylcarnitine were greater losses in the activity of sarcolemmal (Na,K)-ATPase. Since all three kinds of amphiphilic lipids accumulate during ischemia, this study suggests a novel mechanism of potentiation of sacolemmal membrane injury when free radicals are present. MH - Adenosine Triphosphatase, Sodium, Potassium/METABOLISM ; Animal ; Dogs ; Free Radicals ; Fumarates/PHARMACODYNAMICS ; Lipid Peroxides/*METABOLISM ; Lipids/*METABOLISM ; Myocardium/CYTOLOGY/ ENZYMOLOGY ; Oxidation-Reduction ; Palmitoyl Coenzyme A/ PHARMACODYNAMICS ; Palmitoylcarnitine/PHARMACODYNAMICS ; Sarcolemma/*METABOLISM ; Support, U.S. Gov't, P.H.S. SO - J Biol Chem 1986 Jan 25;261(3):1153-7 44 UI - 86103454 AU - Pagonis C ; Tauber AI ; Pavlotsky N ; Simons ER TI - Flavonoid impairment of neutrophil response. AB - Flavonoids are a class of phenolic plant pigments which impair the oxidative burst of neutrophils to an extent dependent on their hydrophobicity. The distribution of quercetin and of morin in nitrogen-cavitated neutrophils paralleled their respective hydrophobic characteristics and respiratory burst inhibition. While both flavonoids were localized primarily in the specific granule membrane of neutrophils, the amount of quercetin was considerably greater than that of morin. We here demonstrate inhibition of the initial stimulation response, depolarization of the membrane potential as monitored by fluorescence of the membrane probe diS-C3-(5), and of the respiratory burst, monitored by following the destruction of diS-C3-(5), a reaction mediated by the H2O2 produced in the burst. The flavonoids kaempferol, morin, quercetin, or fisetin were preincubated with human neutrophils at a concentration of 100 microM per 2 X 10(6) cells/ml for 2-3 min and subsequently stimulated with 1 microgram/ml of the tumor promoter phorbol myristate acetate (PMA) or with 60 micrograms/ml of immune complex. The effect of each compound differed, i.e. depolarization was enhanced by some and inhibited by others, while H2O2 generation was inhibited by each, supporting our previous findings that membrane potential depolarization and the respiratory burst are dissociable events. Concentration-response experiments, performed at flavonoid concentrations between 12.5 and 500 microM to determine the IC50 values of these compounds for depolarization and burst activation, indicated that none of the flavonoids affected the resting potential, while all perturbed the stimulus-coupled response, the direction and extent of the perturbation depending upon the stimulus, and the function assessed. These data show that the effects of flavonoids on human neutrophils are complex and suggest several sites of action depending upon the flavonoid's subcellular distribution and pathway of stimulation. MH - Flavones/*PHARMACODYNAMICS ; Free Radicals ; Human ; Hydrogen Peroxide/METABOLISM ; Membrane Potentials/DRUG EFFECTS ; Neutrophils/*DRUG EFFECTS/METABOLISM ; Oxygen Consumption/DRUG EFFECTS ; Quercetin/PHARMACODYNAMICS ; Solubility ; Superoxide/ BIOSYNTHESIS ; Support, U.S. Gov't, P.H.S. ; Tetradecanoylphorbol Acetate/PHARMACODYNAMICS SO - Biochem Pharmacol 1986 Jan 15;35(2):237-45 45 UI - 86087947 AU - Asayama K ; Dettbarn WD ; Burr IM TI - Differential effect of denervation on free-radical scavenging enzymes in slow and fast muscle of rat. AB - To determine the effect of denervation on the free-radical scavenging systems in relation to the mitochondrial oxidative metabolism in the slow-twitch soleus and fast-twitch extensor digitorum longus (EDL) muscles, the sciatic nerve of the rat was crushed in the midthigh region and the muscle tissue levels of five enzymes were studied 2 and 5 weeks following crush. Recently developed radioimmunoassays were utilized for the selective measurement of cuprozinc (cytosolic) and mangano (mitochondrial) superoxide dismutases. Total tissue content of cuprozinc superoxide dismutase showed a mild decrease after denervation in slow but not in fast muscle. Manganosuperoxide dismutase and fumarase decreased markedly at 2 weeks and returned toward control levels by 5 weeks, the changes appearing to be greater in slow than in fast muscle. At 2 weeks, cytochrome c oxidase decreased significantly in slow, but not in fast muscle. GSH-peroxidase at baseline was 10-fold higher in slow than in fast muscle, markedly decreased at 2 weeks in slow muscle, and returned toward control levels at 5 weeks, whereas the total enzyme activity in fast muscle did not change through 5 weeks. These data represent the first systematic report of free radical scavenging systems in slow and fast muscles in response to denervation. Selective modification of cuprozinc and manganosuperoxide dismutases and differential regulation of GSH-peroxidase was demonstrated in slow and fast muscle. MH - Animal ; Copper ; Cytochrome Oxidase/METABOLISM ; Free Radicals ; Fumarate Hydratase/METABOLISM ; Glutathione Peroxidase/METABOLISM ; Male ; *Muscle Denervation ; Muscles/*ENZYMOLOGY ; Organ Weight ; Rats ; Rats, Inbred Strains ; Superoxide Dismutase/METABOLISM ; Support, U.S. Gov't, Non-P.H.S. ; Zinc SO - J Neurochem 1986 Feb;46(2):604-9 46 UI - 86061722 AU - Koide T ; Asano T ; Matsushita H ; Takakura K TI - Enhancement of ATPase activity by a lipid peroxide of arachidonic acid in rat brain microvessels. AB - The effects of 15-hydroperoxyarachidonic acid (15-HPAA) on Na+, K+- and Mg+-ATPase activities in the blood-brain barrier (BBB) were examined using rat brain microvessels (MV). 15-HPAA markedly stimulated these ATPase activities in MV at low concentrations whereas the synaptosomal Na+, K+-ATPase activity was inhibited in a dose-dependent manner. Further neurochemical analysis revealed that this stimulatory effect of 15-HPAA in MV was not due to a simple detergent-like action of the compound on the membranes but rather to stimulation of the phospholipase A2 and lipoxygenase activity within MV. In addition, it was shown that free radical reactions were involved in the mechanism. Since such anti-edema drugs as 1,2-bis(nicotinamido)propane were proved to be potent suppressors of the enhanced ATPase activity, further speculations on the role of this effect for ischemic brain edema are offered. MH - Adenosine Triphosphatase/*METABOLISM ; Adenosine Triphosphatase, Magnesium/METABOLISM ; Adenosine Triphosphatase, Sodium, Potassium/METABOLISM ; Animal ; Arachidonic Acids/ *PHARMACODYNAMICS ; Brain/*BLOOD SUPPLY/DRUG EFFECTS/ENZYMOLOGY ; Brain Edema/METABOLISM ; Dose-Response Relationship, Drug ; Fatty Acids/PHARMACODYNAMICS ; Free Radicals ; Hydroquinones/ PHARMACODYNAMICS ; Lipid Peroxides/*PHARMACODYNAMICS ; Lipoxygenases/METABOLISM ; Nicotinamide/ANALOGS & DERIVATIVES/ PHARMACODYNAMICS ; Phospholipases A/METABOLISM ; Quinacrine/ PHARMACODYNAMICS ; Rats ; Synaptosomes/DRUG EFFECTS/ENZYMOLOGY ; Vitamin E/PHARMACODYNAMICS SO - J Neurochem 1986 Jan;46(1):235-42 1 UI - 87101104 AU - Thomas MJ ; Shirley PS ; Hedrick CC ; DeChatelet LR TI - Role of free radical processes in stimulated human polymorphonuclear leukocytes. AB - Human polymorphonuclear leukocytes produce large quantities of superoxide when they attack and kill bacteria. However, superoxide is a weak oxidizing and reducing agent, and other more reactive oxygen species derived from reactions of superoxide are suggested to participate in the killing processes. To test the hypothesis that a reactive free radical or singlet oxygen is involved in bactericidal activity, human polymorphonuclear leukocytes were exposed to phagocytozable particles containing lipids that contain the easily autoxidized 1,4-diene moiety. After incubation the preparations were extracted and the extracts reduced with NaBH4 to convert hydroperoxides to stable alcohols. Using gas chromatography/mass spectrometry to analyze the extracts, we were unable to detect products unless iron salts were added to the medium. The products obtained by extraction are those that would be expected if both free radical chain autoxidation and 1O2 oxidation were taking place. In summary, we find that polymorphonuclear leukocytes do not cause peroxidation, implying that formation of strongly oxidizing free radicals is not an intrinsic property of the leukocyte. Added iron catalyzes peroxidation by activated leukocytes yielding an unusual distribution of hydroxylated products. MH - Free Radicals ; Human ; Hydrogen Peroxide/BLOOD ; Mass Fragmentography ; Neutrophils/*PHYSIOLOGY ; Oxidation-Reduction ; Oxygen Consumption ; Oxygen/BLOOD ; Phagocytosis ; Superoxide/ *BLOOD ; Support, U.S. Gov't, P.H.S. SO - Biochemistry 1986 Dec 2;25(24):8042-8 2 UI - 87092581 AU - Tahamont MV ; Gee MH TI - The effects of cyclooxygenase inhibition on chemiluminescence and aggregation in sheep neutrophils. AB - Antiinflammatory actions of cyclooxygenase inhibitors may be related to inhibition of the synthesis and release of prostaglandins and thromboxane or to nonspecific actions of particular drugs. The role of cyclooxygenase products of arachidonic acid was studied in two leukocyte functions, free radical release and aggregation, after complement activation. Dose response curves were constructed after treatment with meclofenamate or ibuprofen. To differentiate between effects on free radical release from complement activated neutrophils and scavenging free radicals, additional experiments were made with a cell free system to generate free radicals. Both drugs inhibited complement initiated neutrophil chemiluminescence in a dose dependent manner. Meclofenamate acted primarily as a scavenger while ibuprofen inhibited free radical release. Neither drug had any inhibitory effects on complement induced leukocyte aggregation. MH - Animal ; Cell Aggregation/DRUG EFFECTS ; Complement Activation ; Free Radicals ; Ibuprofen/PHARMACODYNAMICS ; In Vitro ; Luminescence ; Meclofenamic Acid/PHARMACODYNAMICS ; Neutrophils/ DRUG EFFECTS/IMMUNOLOGY/*PHYSIOLOGY ; Prostaglandin Synthase/ *ANTAGONISTS & INHIBITORS ; Sheep ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Prostaglandins Leukotrienes Med 1986 Oct;24(2-3):139-49 3 UI - 87076674 AU - Vissers MC ; Winterbourn CC TI - The effect of oxidants on neutrophil-mediated degradation of glomerular basement membrane collagen. AB - The contribution of activated oxygen species to neutrophil-mediated degradation of basement membrane collagen was investigated. In preliminary experiments, pre-exposure of either albumin or glomerular basement membrane to neutrophil myeloperoxidase with H2O2 and chloride increased their susceptibility to proteolysis 2-3-fold. In the basement membrane model, neutrophils are stimulated by trapped immune complexes to adhere, produce oxidants and degranulate. Degradation, measured as the amount of hydroxyproline solubilised, was due to neutral proteinases, particularly elastase, and depended on cell number and the amount of proteinase released. Experiments with oxidant scavengers and inhibitors and with neutrophils from donors with chronic granulomatous disease or myeloperoxidase deficiency showed that oxidants did not affect degradation of the basement membrane when this was measured on a per cell basis. However, oxidative inactivation of the released granule enzymes occurred. Activities of elastase, beta-glucuronidase and lysozyme were 1.5-2-times higher in the presence of catalase, but were unaffected by superoxide dismutase or hydroxyl radical scavengers. Inactivation did not occur with chronic granulomatous disease or myeloperoxidase deficient neutrophils. When related to the activity of released elastase, or to other degranulation markers, collagen degradation was decreased in the presence of catalase, or with chronic granulomatous disease or myeloperoxidase deficient cells. This implies that the basement membrane was made more digestible by myeloperoxidase-derived oxidants, as occurred in the cell-free experiments. Taken together, the results indicate that neutrophil oxidants have two opposing effects. They increase the susceptibility of the collagen to proteolysis and inactivate the proteinases responsible. MH - Basement Membrane/*METABOLISM ; Chlorides/*PHARMACODYNAMICS ; Collagen/*METABOLISM ; Cytoplasmic Granules/ENZYMOLOGY/SECRETION ; Free Radicals ; Granulomatous Disease, Chronic/PATHOLOGY ; Human ; Hydrogen Peroxide/*PHARMACODYNAMICS ; IgG ; Kidney Glomerulus/*DRUG EFFECTS ; Myeloperoxidase/DEFICIENCY/METABOLISM ; Neutrophils/*DRUG EFFECTS/ENZYMOLOGY/SECRETION ; Oxidation-Reduction ; Peptide Hydrolases/METABOLISM/SECRETION ; Support, Non-U.S. Gov't SO - Biochim Biophys Acta 1986 Dec 19;889(3):277-86 4 UI - 87058202 AU - Asman B ; Bergstr:om K ; Wijkander P ; Lockowandt B TI - Influence of plasma components on luminol-enhanced chemiluminescence from peripheral granulocytes in juvenile periodontitis. AB - The generation rate of free oxygen radicals as measured by maximal light intensity of luminol-enhanced chemiluminescence from peripheral blood granulocytes (PMN) stimulated with differently opsonized Staphylococcus aureus was studied in 13 patients with juvenile periodontitis (JP) and pair-matched, healthy controls. Plasma proteins related to inflammation were also assayed. When stimulated with bacteria opsonized with autologous serum, the PMN from the JP patients showed a more intensive chemiluminescence than did their pair-matched controls (p less than or equal to 0.0005). The difference was consistent but slightly reduced when using heat-treated serum (p less than or equal to 0.006) or heterologous gammaglobulin (p less than or equal to 0.19) for opsonization. When testing freeze preserved sera from 11 of the compared pairs, the sera from JP patients induced a slightly higher chemiluminescence in PMN from a healthy donor (p less than or equal to 0.031). Protein analysis of the patient sera revealed a slightly higher concentration of complement 4 (p less than or equal to 0.032) and IgM (p less than or equal to 0.030) when compared with their respective pair-matched healthy controls. The influence of other blood components contaminating our assay system was checked on healthy PMN cells. Lymphocytes, platelets, relevant amounts of ADP and serum had no effect on the chemiluminescence. In conclusion, the increased chemiluminescence of peripheral blood granulocytes from patients with juvenile periodontitis seems to be related mainly to the cells. The association with free oxygen radicals and their tissue-damaging potency might be a contributing factor in the pathogenesis of periodontal disease. MH - Adolescence ; Adult ; Blood Proteins/PHYSIOLOGY ; Female ; Free Radicals ; Granulocytes/*METABOLISM ; Human ; *Luminescence ; Luminol/*PHARMACODYNAMICS ; Male ; Neutrophils/*METABOLISM ; Opsonins/PHARMACODYNAMICS ; Oxygen ; Periodontal Diseases/*BLOOD ; Periodontosis/*BLOOD ; Phagocytosis ; Pyridazines/ *PHARMACODYNAMICS SO - J Clin Periodontol 1986 Oct;13(9):850-5 5 UI - 87056225 AU - Zeis BM ; Anderson R TI - Clofazimine-mediated stimulation of prostaglandin synthesis and free radical production as novel mechanisms of drug-induced immunosuppression. AB - The effects of clofazamine (3-(p-chloroanilino)-10-(p-chlorophenyl)-2, 10-dihydro-2-(isopropylimino)-phenazine) at concentrations of 0.625-20 micrograms/ml on the mitogen-induced transformation, luminol-enhanced chemiluminescence, arachidonic acid metabolism and sulphydryl content of human mononuclear leucocytes (MNL) were investigated in vitro. The drug at all concentrations tested decreased MNL sulphydryl content and inhibited mitogen-induced transformation. Clofazimine increased the spontaneous luminol-enhanced chemiluminescence and activated the arachidonic acid cascade in MNL. The anti-oxidants ascorbic acid and cysteine and the prostaglandin (PG) synthesis inhibitor indomethacin were used individually and in combination to identify the primary mediators of the anti-proliferative effects of clofazimine on MNL. Combinations of an anti-oxidant with a PG synthesis inhibitor completely protected MNL from clofazimine mediated inhibition of mitogen-induced transformation. These results show that the anti-proliferative activity of clofazimine is related to both the pro-oxidative and PG synthesis enhancing effects of the drug on MNL. MH - Clofazimine/*PHARMACODYNAMICS ; Free Radicals ; Human ; *Immunosuppression ; In Vitro ; Luminescence ; Mitogens/ PHARMACODYNAMICS ; Models, Biological ; Neutrophils/DRUG EFFECTS ; Phytohemagglutinins/PHARMACODYNAMICS ; Prostaglandins E/ BIOSYNTHESIS ; Prostaglandins/*BIOSYNTHESIS ; Sulfhydryl Compounds/METABOLISM ; Support, Non-U.S. Gov't ; SRS-A/ BIOSYNTHESIS ; Thromboxane B2/BIOSYNTHESIS SO - Int J Immunopharmacol 1986;8(7):731-9 6 UI - 87052290 AU - Bron AJ ; Cheng H TI - Cataract and retinopathy: screening for treatable retinopathy. AB - Diabetes causes cataract and certain physical changes in the lens. The diabetic lens is larger than the non-diabetic and shows greater light scatter and fluorescence. Both hyperglycaemia and lowering of blood glucose case refractive changes and hypermetropia is the most common. Classical 'snow-flake' juvenile cataract associated with hyperglycaemia is now rare. It has an osmotic mechanism. Diabetes is a risk factor for cataract in adults which is duration dependent, more frequent in women and leads to earlier surgery. It resembles non-diabetic senile cataract. Extracapsular cataract extraction is the method of choice for diabetic cataract with a better visual result and less risk of rubeosis iridis. A posterior chamber implant may still permit retinal photocoagulation if necessary. Diabetic retinopathy is still the leading cause of blindness in the working age group. The beneficial effect of photocoagulation has been shown by randomized controlled trials to be long-lasting for both proliferative retinopathy and maculopathy. Therefore there is a need for screening, especially for those with proliferative disease which may be present without symptoms. A knowledge of risk factors will enhance detection rate with duration as the strongest determinant for retinopathy. Any screening modality should be highly sensitive as well as specific. The role of different professionals as potential screeners should be considered. Adequate provisions include facilities for checking vision and for dimming ambient lighting. Mydriasis and a good ophthalmoscope light will increase detection rate. The use of a 45 degrees non-mydriatic camera is unlikely to supplant the use of an ophthalmoscope as a single field is likely to miss important lesions. A 60 degrees camera may confer a large enough field and the use of transparencies will provide magnification when films are projected but the camera is more difficult to use. A list of features chosen by a recent study to characterize sight-threatening retinopathy is included and their presence indicates the need for referral to an ophthalmic clinic for treatment or close observation. MH - Adolescence ; Adult ; Aldose Reductase/ANTAGONISTS & INHIBITORS ; Cataract Extraction ; Cataract/*DIAGNOSIS/PATHOLOGY ; Diabetes Mellitus, Insulin-Dependent/PATHOLOGY ; Diabetes Mellitus, Non-Insulin-Dependent/PATHOLOGY ; Diabetic Retinopathy/*DIAGNOSIS/ PATHOLOGY/THERAPY ; Enzyme Inhibitors/THERAPEUTIC USE ; Female ; Free Radicals ; Glycosylation ; Human ; Lens, Crystalline/ PATHOLOGY/PHYSIOPATHOLOGY ; Male ; Mass Screening ; Middle Age ; Photography ; Review SO - Clin Endocrinol Metab 1986 Nov;15(4):971-99 7 UI - 87036674 AU - Burt HM ; Evans E ; Lam EW ; Gehrs PF ; Herring FG TI - Membranolytic effects of monosodium urate monohydrate: influence of grinding. AB - The effect of grinding on the membranolytic interaction between monosodium urate monohydrate (MSUM) crystals and intact erythrocyte membranes was studied. Crystals were ground for between 1-72 h, and percent hemolysis and zeta potentials determined. A cationic amphiphilic probe (CAT12) was incorporated into the erythrocyte membrane and incubated with MSUM. Increasing grinding times caused a decrease in both the crystallinity and zeta potential of the samples, a decrease in percent hemolysis values and a change in the distribution of free and bound spin label populations. The probe redistribution is thought to be due to an electrostatic interaction between negatively charged MSUM and the CAT12 probe. MH - Crystallization ; Electron Spin Resonance ; Erythrocyte Membrane/ *DRUG EFFECTS ; Hemolysis/DRUG EFFECTS ; Human ; In Vitro ; Inflammation/CHEMICALLY INDUCED ; Powders ; Spin Labels ; Support, Non-U.S. Gov't ; Uric Acid/ISOLATION & PURIFICATION/ *TOXICITY ; X-Ray Diffraction SO - J Rheumatol 1986 Aug;13(4):778-83 8 UI - 87030684 AU - Raphael GD ; Metcalfe DD TI - Mediators of airway inflammation. AB - Mediators of inflammation can be classified into those that exist preformed in cells and those that are rapidly generated following stimulation. Vasoactive amines, lysosomal enzymes, neuropeptides and proteoglycans are examples of classes of mediators that are synthesized and stored, awaiting the proper stimulus that results in their release. Prostaglandins, leukotrienes, platelet activating factors and free radicals are representative substances that are generated as a result of cell activation. It is now clear that similar inflammatory mediators can be produced by diverse cell types. The exact role of specific mediators of inflammation depends on several factors: the precise stimulus that results in their release or production, the time in the overall inflammatory process when they are produced, and their specific biological effects. Upon stimulation, resident cells such as epithelial cells, mast cells and macrophages are in a position to induce inflammation, whereas cells entering sites of initial injury are able to perpetuate the process. Specificity further depends upon cell surface receptors and the mediators resulting from receptor perturbation. Finally, the most critical question is how inflammation, once induced, is limited by the mammalian system in order to prevent this process from unlimited extension. It would appear that numerous mechanisms which limit injury are brought into play with the induction of inflammation. These mechanisms include degradation of mediators by other mediators, mediator degradation by resident cells, and mediator dilution by tissue fluids. In summary, the role of mediators is extremely complex, involving their innate properties, temporal expression, and inactivation. MH - Arachidonic Acids/METABOLISM ; Free Radicals ; Histamine Liberation ; Human ; Immunity, Cellular ; Inflammation/ *PHYSIOPATHOLOGY ; Mast Cells/PHYSIOLOGY ; Membrane Lipids/ PHYSIOLOGY ; Models, Biological ; Platelet Activating Factor/ PHYSIOLOGY ; Prostaglandins/PHYSIOLOGY ; Respiratory Tract Infections/*PHYSIOPATHOLOGY ; Review SO - Eur J Respir Dis [Suppl] 1986;147:44-56 9 UI - 87028675 AU - Cavarocchi NC ; England MD ; Schaff HV ; Russo P ; Orszulak TA ; Schnell WA Jr ; O'Brien JF ; Pluth JR TI - Oxygen free radical generation during cardiopulmonary bypass: correlation with complement activation. AB - To determine the relationships among complement activation, pulmonary leukosequestration, and oxygen free radical generation, we prospectively studied 15 patients undergoing cardiopulmonary bypass for myocardial revascularization. Plasma levels of C3a, C4a, and hydrogen peroxide (a marker of oxygen free radical generation) were measured before, during, and after extracorporeal circulation. The results confirm that cardiopulmonary bypass activates complement via the alternate (C3a) pathway. This first phase of complement activation was accompanied by an increase in plasma H2O2 (from 80 +/- 8 to 155 +/- 13 microM/ml; p less than .001) and by pulmonary sequestration of polymorphonuclear leukocytes. Protamine administration after cardiopulmonary bypass further activated complement via the classical (C4a) pathway but was not accompanied by a change in plasma hydrogen peroxide. We hypothesize that both complement activation and excess oxygen free radical generation contribute to the pathophysiology of extracorporeal circulation. MH - Blood Cells/PATHOLOGY ; *Cardiopulmonary Bypass ; Cell Count ; *Complement Activation ; Complement/ANALYSIS ; Extracorporeal Circulation ; Free Radicals ; Human ; Hydrogen Peroxide/BLOOD ; Leukocytes/PATHOLOGY ; *Oxygen ; Pulmonary Circulation SO - Circulation 1986 Nov;74(5 Pt 2):III130-3 10 UI - 87026584 AU - Kunicki TJ ; Nugent DJ ; Piotrowicz RS ; Lai CS TI - Covalent attachment of sulfhydryl-specific, electron spin resonance spin-labels to Fab' fragments of murine monoclonal antibodies that recognize human platelet membrane glycoproteins. Development of membrane protein specific spin probes. AB - A general method for the production of high-affinity, nitroxide-labeled, protein-specific spin probes is described in this paper. Fab' fragments are generated from protein-specific, murine monoclonal antibodies by pepsin digestion and mild reduction with cysteine. The free sulfhydryl group located in the carboxy-terminal region of these molecules and produced de novo by this manipulation is then alkylated by reaction with 4-maleimido-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO-maleimide), thereby generating spin-labeled Fab' fragments of these monoclonal antibodies. Two prototypic monoclonal antibodies were tested, each specific for a different integral membrane glycoprotein of human blood platelets. The results indicate that Fab' spin probes generated by this method retain the ability to bind to these glycoproteins within the membrane of intact platelets. These reagents thus represent probes that can be generally used to monitor integral membrane protein mobility on the surface of the intact cell. MH - Animal ; *Antibodies, Monoclonal/ISOLATION & PURIFICATION ; Antigen-Antibody Complex ; Blood Platelets/*ANALYSIS ; Cell Membrane/ANALYSIS ; Cyclic N-Oxides ; Electron Spin Resonance ; Glycoproteins/*BLOOD ; Human ; IgG/ISOLATION & PURIFICATION ; *Immunoglobulins, Fab/ISOLATION & PURIFICATION ; Membrane Proteins/*BLOOD ; Mice ; Spin Labels ; Sulfhydryl Compounds/ ANALYSIS ; Support, U.S. Gov't, P.H.S. SO - Biochemistry 1986 Sep 9;25(18):4979-83 11 UI - 87026056 AU - Biemond P ; Swaak AJ ; van Eijk HG ; Koster JF TI - Intraarticular ferritin-bound iron in rheumatoid arthritis. A factor that increases oxygen free radical-induced tissue destruction. AB - Iron mobilized from ferritin is able to convert superoxide and hydrogen peroxide, which are produced in large amounts in rheumatoid arthritis (RA), to the extremely toxic hydroxyl radical. We have found that synovial fluid ferritin is increased significantly in RA patients compared with levels in controls. The high synovial fluid:serum ferritin ratio is compatible with the hypothesis that synovial fluid ferritin is derived from the synovial membrane. We found no difference in ferritin concentrations in the synovial membranes of RA patients compared with those of controls. Quantitative data on the amount of iron bound to ferritin showed that the level was 2.9 times higher in RA synovial membranes than in those of controls. Moreover, RA synovial fluid contained considerable amounts of iron bound to ferritin. Calculation of the iron saturation of ferritin revealed that RA synovial membranes contained a mean of 2,210 moles of iron per mole of ferritin: a significant elevation when compared with the mean value of 1,500 moles found in the synovial membranes of the controls. The decreased saturation of ferritin in RA synovial fluid, compared with that in the synovial membrane, could be caused by an uncompensated release of iron from ferritin, which has been induced by superoxide that is produced by stimulated granulocytes. The results demonstrate that in the joints of RA patients, sufficient ferritin loaded with iron is available to stimulate oxygen free radical damage. MH - Arthritis, Rheumatoid/BLOOD/*METABOLISM ; Ferritin/*METABOLISM ; Free Radicals ; Human ; Iron/*METABOLISM ; Joints/*METABOLISM ; Knee Injuries/BLOOD/METABOLISM ; Osteoarthritis/BLOOD/METABOLISM ; Protein Binding ; Support, Non-U.S. Gov't ; Synovial Fluid/ METABOLISM ; Synovial Membrane/METABOLISM SO - Arthritis Rheum 1986 Oct;29(10):1187-93 12 UI - 87004435 AU - Kasai H ; Nishimura S TI - Hydroxylation of guanine in nucleosides and DNA at the C-8 position by heated glucose and oxygen radical-forming agents. AB - Heated glucose is mutagenic to Salmonella typhimurium TA 100 in the absence of S-9 mix. For identifying unknown mutagens in heated glucose (dry solid, 200 degrees C, 20 min), reaction with isopropylideneguanosine (IPG) was followed by isolation and characterization of the mutagen-IPG adduct. Two adducts, glyoxal-IPG and 8-hydroxy-IPG, were identified in the reaction mixture by this technique. To elucidate the mechanism of this hydroxylation reaction, we investigated the abilities of various agents to hydroxylate deoxyguanosine or guanine base in DNA. Various reducing agents, metals, asbestoses, polyphenols, aminophenols, and X-ray were effective for hydroxylation, and an oxygen radical seems to be the reactive species. For sensitive detection of 8-hydroxyguanine, a monoclonal antibody for it was prepared. MH - Antibodies, Monoclonal ; Deoxyguanosine/ANALOGS & DERIVATIVES/ IMMUNOLOGY ; DNA ; *Food Analysis ; Food Contamination ; Free Radicals ; *Glucose ; *Guanine ; Heat ; Hydroxylation ; Mutagens/ *ISOLATION & PURIFICATION ; Nucleosides ; Oxygen ; Support, Non-U.S. Gov't SO - Environ Health Perspect 1986 Aug;67:111-6 13 UI - 87002161 AU - Goodglick LA ; Kane AB TI - Role of reactive oxygen metabolites in crocidolite asbestos toxicity to mouse macrophages. AB - Crocidolite asbestos is toxic to macrophages in vitro. We hypothesize that this toxicity is mediated by the generation of reactive oxygen metabolites. Elicited mouse peritoneal macrophages were found to release reactive oxygen metabolites upon incubation with crocidolite asbestos in vitro. Crocidolite toxicity to both primary cultures of mouse peritoneal macrophages and P388D1 cells, a mouse macrophage-like cell line, could be prevented by a hypoxic environment or by addition of the reactive oxygen metabolite scavengers, superoxide dismutase and catalase. In addition, if crocidolite fibers were presoaked with the iron chelator deferoxamine, no macrophage death occurred. In an attempt to mimic crocidolite-induced cytotoxicity, P388D1 cells or primary elicited macrophages were exposed to the nontoxic mineral particle titanium dioxide in the presence and absence of ferric chloride. Titanium dioxide was only lethal when ferric chloride was added. This toxicity was prevented by superoxide dismutase, catalase, or deferoxamine. These results suggest that crocidolite-induced injury to macrophages depends on the formation of reactive oxygen metabolites. Iron present in crocidolite fibers may catalyze the production of hydroxyl radical from superoxide anion and hydrogen peroxide generated during phagocytosis. These highly reactive hydroxyl radicals are postulated to mediate lethal cell injury. MH - Animal ; Asbestos/*TOXICITY ; Catalase/METABOLISM ; Cell Line ; Cell Survival/DRUG EFFECTS ; Deferoxamine/PHARMACODYNAMICS ; Ferric Compounds ; Free Radicals ; Hydrogen Peroxide/TOXICITY ; In Vitro ; Macrophages/*DRUG EFFECTS ; Mice ; Oxygen/*TOXICITY ; Phagocytosis ; Superoxide Dismutase/METABOLISM ; Superoxide/ TOXICITY ; Support, U.S. Gov't, P.H.S. ; Titanium/TOXICITY SO - Cancer Res 1986 Nov;46(11):5558-66 14 UI - 87000766 AU - Morli:ere P TI - Drug-induced photosensitivity: phototoxic and photoallergic reactions--a few molecular aspects. AB - Drug-induced photosensitivity involves mainly phototoxic and photoallergic reactions. The main features of phototoxic and photoallergic reactions are presented and some molecular aspects involved in the mechanisms leading to an adverse skin response are illustrated with examples. MH - Animal ; Antigens/IMMUNOLOGY ; Chlordiazepoxide/ADVERSE EFFECTS ; DNA ; Free Radicals ; Human ; Light ; Oxygen ; Phenothiazines/ ADVERSE EFFECTS ; Photochemistry ; Photosensitivity Disorders/ *CHEMICALLY INDUCED/DIAGNOSIS/IMMUNOLOGY ; Propionic Acids/ ADVERSE EFFECTS ; Psoralens/ADVERSE EFFECTS ; Review ; Sulfonamides/ADVERSE EFFECTS ; Superoxide ; Time Factors ; Ultraviolet Rays SO - Biochimie 1986 Jun;68(6):849-55 15 UI - 87000567 AU - Berliner LJ ; Musci G ; Maliarik M ; Plessas NR ; Goldstein IJ TI - Binding of N-acetylgalactosamine-specific lectins to spin-labeled galactosamine derivatives. AB - Legume seed lectins specific for N-acetyl-alpha-D-galactosaminyl end groups from Amphicarpaea bracteata, lima bean, Griffonia simplicifolia, Dolichos biflorus, and soybean were compared with respect to binding of several spin-labeled derivatives of D-galactosamine by electron spin resonance and precipitin inhibition analysis. Spin-label II [methyl 2-[[(2,2,5,5-tetramethyl-1-oxopyrrolidin-3-yl) carbonyl]amino]-2-deoxy-alpha-D-galactopyranoside], spin-label III [1-(methyl 2-deoxy-alpha-D-galactopyranosid-2-yl)-3-(2,2,6, 6-tetramethyl-1-oxypiperidin-4-yl)-2-thiourea], and spin-label IV [1-[4-[[(methyl 2-deoxy-alpha-D-galactopyranosid-2-yl)amino]carbonyl]phenyl]-3-(2- , 2,6-tetramethyl-1-oxypiperidin-4-yl)-2-thiourea] contain 2-N-(oxypiperidinyl) or 2-N-(oxypyrrolidinyl) substituents varying in length and polarity of the linker arm between the glycoside and nitroxide ring. Spin-labels II and III were found to bind very weakly to all the lectins tested (Kd greater than or equal to 1.0 mM). Spin-label IV, containing a planar, nonpolar 2-N-phenyl group, was bound very strongly (Kd = 0.1-0.4 mM) and was moderately immobilized (2T parallel = 48-56 G) by all lectins except that from D. biflorus. Notably, the affinity of spin-label IV to lima bean lectin was 18-fold greater than that for methyl N-acetyl-alpha-galactosaminide. These results suggest that when the bulky oxypiperidinyl moiety lies in a position close to the sugar ring, it interferes with binding; in the cases where a phenyl group spacer exists, the aromatic ring in some cases actually enhances binding.(ABSTRACT TRUNCATED AT 250 WORDS) MH - *Acetylgalactosamine ; Comparative Study ; Electron Spin Resonance ; *Galactosamine/ANALOGS & DERIVATIVES ; *Lectins ; *Spin Labels ; Structure-Activity Relationship ; Support, U.S. Gov't, P.H.S. SO - Biochemistry 1986 Jul 29;25(15):4457-61 16 UI - 86306105 AU - Ghezzi P ; Saccardo B ; Bianchi M TI - Induction of xanthine oxidase and heme oxygenase and depression of liver drug metabolism by interferon: a study with different recombinant interferons. AB - Induction of xanthine oxidase in mouse liver by interferon (IFN) was studied with three different recombinant human leukocyte IFN molecules: IFLrA, IFLrD and the hybrid IFLrA/D(Bgl II). The ability of different IFN species to induce xanthine oxidase correlated with their ability to depress liver cytochrome P-450-dependent drug metabolism, supporting the hypothesis that reactive oxygen metabolites generated by xanthine oxidase might be responsible for this impairment of liver function by IFN. The antioxidant N-acetylcysteine protected in vivo against the depression of liver drug metabolism by IFLrA/D. IFLrA/D was also found to induce liver microsomal heme oxygenase, an effect that was probably secondary to the observed depression of cytochrome P-450. MH - Animal ; Cytochrome P-450/METABOLISM ; Drugs/METABOLISM ; Enzyme Induction/DRUG EFFECTS ; Free Radicals ; Heme Oxygenase/ *BIOSYNTHESIS ; Hydroxylases/*BIOSYNTHESIS ; Interferon Type I/ *PHARMACODYNAMICS ; Liver/*DRUG EFFECTS/METABOLISM ; Male ; Mice ; Superoxide/METABOLISM ; Support, Non-U.S. Gov't ; Xanthine Oxidase/*BIOSYNTHESIS SO - J Interferon Res 1986 Jun;6(3):251-6 17 UI - 86149368 AU - Pontremoli S ; Melloni E ; Michetti M ; Sacco O ; Sparatore B ; Salamino F ; Damiani G ; Horecker BL TI - Cytolytic effects of neutrophils: role for a membrane-bound neutral proteinase. AB - A neutral serine proteinase, purified 250-fold from the plasma membrane fraction of human neutrophils, differs in its catalytic and molecular properties from the well-known neutral proteinases present in azurophil (primary) granules. Stimulation of neutrophils with low concentrations of phorbol 12-myristate 13-acetate (PMA) results in the release into the medium of the membrane-bound proteinase and the concomitant production of oxygen radicals. These concentrations of PMA also induce full cytolytic activity measured with 51Cr-labeled ox erythrocytes. A role for the neutral serine proteinase in the cytolytic activity of PMA-stimulated neutrophils is supported by the following observations: (i) the lytic activity of the stimulated neutrophils is correlated with the quantity of neutral proteinase present in the membranes; (ii) the extracellular medium from PMA-stimulated neutrophils causes the cytolysis of 51Cr-labeled erythrocytes that have been exposed to nonlytic concentrations of H2O2; (iii) cytolysis of H2O2-treated erythrocytes is also observed with the crude proteinase solubilized from neutrophil membranes or with the purified proteinase from the same source; and (iv) in each case the cytolytic activity is proportional to the proteinase activity present and is prevented by the addition of serine proteinase inhibitors. We conclude that cytolysis of target cells by PMA-activated neutrophils can result from the cooperative effects of oxygen radicals and the membrane-bound neutral serine proteinase. The participation of enzymes from specific (secondary) granules is excluded because, with the low concentrations of PMA employed, very little release of secondary granule constituents is observed. MH - Cytotoxicity, Immunologic ; Erythrocytes/DRUG EFFECTS ; Free Radicals ; Human ; Hydrogen Peroxide/PHARMACODYNAMICS ; Membrane Proteins/ISOLATION & PURIFICATION/*PHYSIOLOGY/SECRETION ; Neutrophils/DRUG EFFECTS/*ENZYMOLOGY/PHYSIOLOGY ; Oxygen/ METABOLISM ; Peptide Peptidohydrolases/ISOLATION & PURIFICATION/ *PHYSIOLOGY/SECRETION ; Protease Inhibitors/PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Tetradecanoylphorbol Acetate/ PHARMACODYNAMICS SO - Proc Natl Acad Sci USA 1986 Mar;83(6):1685-9 18 UI - 86087182 AU - Weiss SJ ; Curnutte JT ; Regiani S TI - Neutrophil-mediated solubilization of the subendothelial matrix: oxidative and nonoxidative mechanisms of proteolysis used by normal and chronic granulomatous disease phagocytes. AB - Both normal and chronic granulomatous disease (CGD) neutrophils were able to degrade the subendothelial matrix secreted by human endothelial cells via an elastase-dependent process. In the absence of the plasma antiproteinase, alpha-1-proteinase inhibitor (alpha-1-PI), normal neutrophils protect their released elastase from inactivation by using the chlorinated oxidants hypochlorous acid and endogenous N-chloroamines to suppress the antiproteinase's activity. In contrast, CGD neutrophils were unable to generate either class of chlorinated oxidant or to inactivate the porcine pancreatic elastase inhibitory capacity of alpha-1-PI unless the cells were supplemented with exogenous hydrogen peroxide. Despite the reliance of normal neutrophils on chlorinated oxidants to inactivate alpha-1-PI, neutrophils triggered in the presence of agents that block the generation of these reactive species continued to degrade the subendothelial matrix at a suppressed but significant rate in the presence of a 50-fold excess of the antiproteinase. The continued solubilization of the matrix by normal neutrophils was not due to the incomplete inhibition of oxidant generation because triggered CGD neutrophils were also able to degrade the matrix in the presence of excess alpha-1-PI. If CGD neutrophils were stimulated in the presence of an exogenous source of H2O2 and alpha-1-PI, the proteolytic potential of the cells was identical to that observed with normal stimulated neutrophils. We conclude that normal neutrophils can enhance their ability to degrade the subendothelial matrix by oxidatively protecting elastase from inactivation by alpha-1-PI but both normal and CGD neutrophils possess non-oxidatively linked mechanisms for sequestering and using elastase to mediate proteolytic effects in the presence of native antiproteinase. MH - Blood Proteins/METABOLISM/*PHARMACODYNAMICS ; Endothelium/ *METABOLISM ; Enzyme Activation/DRUG EFFECTS ; Extracellular Matrix/METABOLISM ; Free Radicals ; Granulomatous Disease, Chronic/*METABOLISM/PATHOLOGY ; Human ; Male ; Neutrophils/ ENZYMOLOGY/*METABOLISM/PHYSIOLOGY ; Oxygen/*METABOLISM ; Protease Inhibitors/*PHARMACODYNAMICS ; Solubility ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Umbilical Veins SO - J Immunol 1986 Jan;136(2):636-41 19 UI - 86305992 AU - Ranadive NS ; Shirwadkar S ; Persad S ; Menon IA TI - Effects of melanin-induced free radicals on the isolated rat peritoneal mast cells. AB - Pheomelanin from human red hair (RHM) produces considerably more cellular damage in Ehrlich ascites carcinoma cells when subjected to radiations of wavelength 320-700 nm than eumelanin from black hair (BHM). Irradiation of RHM generated large amounts of superoxide while BHM did not produce detectable amounts of superoxide. The present investigations describe the effects of irradiation of mast cells in the presence of various natural and synthetic melanins. Irradiation of mast cells in the presence of RHM and red hair melanoprotein released large amounts of histamine while BHM and synthetic melanins prepared from dopa, cysteinyldopa, or a mixture of dopa and cysteinyldopa did not release histamine. The release of histamine at lower concentrations of RHM was not accompanied by the release of 51Cr from chromium-loaded cells, suggesting that this release was of noncytotoxic nature. On the other hand, the release of histamine at higher concentrations of RHM was due to cell lysis since both histamine and cytoplasmic marker 51Cr were released to the same extent. The release evoked by large concentration RHM was not inhibited by superoxide dismutase or catalase. This suggests that the cell lysis under these conditions was not due to H2O2 or O-2. The finding that mast cells release histamine when irradiated in the presence of RHM suggests that the immediate and late-phase reactions seen in sunburn may in part be due to the release of mediators from these cells. MH - Animal ; Free Radicals ; Histamine Liberation/DRUG EFFECTS ; In Vitro ; Male ; Mast Cells/*METABOLISM/RADIATION EFFECTS ; Melanins/ANALYSIS/*PHARMACODYNAMICS/RADIATION EFFECTS ; Photosensitivity Disorders/ETIOLOGY ; Rats ; Rats, Inbred Strains ; Superoxide/METABOLISM ; Support, Non-U.S. Gov't SO - J Invest Dermatol 1986 Mar;86(3):303-7 20 UI - 86304924 AU - Hatherill JR ; Till GO ; Bruner LH ; Ward PA TI - Thermal injury, intravascular hemolysis, and toxic oxygen products. AB - Acute thermal injury of rat skin produces an early, acute hemoglobinemia that is associated with the presence in blood of osmotically fragile red cells (RBC) that do not contain on their surfaces measurable amounts of complement components. The hemoglobinemia and the appearance in blood of osmotically fragile RBC appear to be the result of complement activation, which leads to oxygen radical production by neutrophils and damage of RBC. This has been demonstrated in vitro as well as in vivo by the ability of antioxidant interventions or neutrophil or complement depletion procedures to prevent the appearance of osmotically fragile RBC and the release of hemoglobin. These data may be relevant to the complications of hemoglobinemia and hemoglobinuria accompanying thermal injury in humans. MH - Animal ; Burns/*BLOOD ; Complement/METABOLISM ; Complement Activation ; Erythrocytes/METABOLISM ; Free Radicals ; *Hemolysis ; In Vitro ; Kinetics ; Male ; Neutrophils/METABOLISM ; Osmotic Fragility ; Oxygen/*BLOOD ; Rats ; Spectrophotometry ; Support, U.S. Gov't, P.H.S. SO - J Clin Invest 1986 Sep;78(3):629-36 21 UI - 86298459 AU - Cleary SF ; Marciano-Cabral F TI - Soluble amoebicidal factors mediate cytolysis of Naegleria fowleri by activated macrophages. AB - Murine peritoneal macrophages activated in vivo with Corynebacterium parvum or bacille Calmette-Gu:erin, in contrast to resident macrophages, demonstrated significant cytolysis of the amoeba, Naegleria fowleri. Catalase and superoxide dismutase, both alone and in combination, failed to inhibit cytolysis of amoebae. N. fowleri amoebae demonstrated significant resistance to exogenously added hydrogen peroxide. The hydroxyl radical scavengers mannitol, thiourea, and dimethyl sulfoxide, as well as anaerobic conditions, failed to inhibit the amoebicidal activity of activated macrophages. Actinomycin D, cycloheximide, and puromycin blocked macrophage amoebicidal activity. Conditioned medium (CM) from lipopolysaccharide-stimulated, but not unstimulated, cultures of activated macrophages was capable of mediating cytolysis of N. fowleri amoebae. Cytolytic activity was recovered by ammonium sulfate precipitation of CM. Heat treatment of the CM inactivated cytolytic activity. Results indicate soluble proteins of activated macrophage origin to be responsible for the amoebicidal activity. MH - Amoeba/*IMMUNOLOGY ; Anaerobiosis ; Animal ; Culture Media ; *Cytotoxicity, Immunologic/DRUG EFFECTS ; Cytotoxins/ANALYSIS/ *PHYSIOLOGY ; Female ; Fractional Precipitation ; Free Radicals ; Heat ; Hydrogen Peroxide/ANTAGONISTS & INHIBITORS/ PHARMACODYNAMICS ; Hydroxides/ANTAGONISTS & INHIBITORS ; *Macrophage Activation/DRUG EFFECTS ; Mice ; Mice, Inbred C57BL ; Proteins/BIOSYNTHESIS ; RNA/BIOSYNTHESIS ; Superoxide/ANTAGONISTS & INHIBITORS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Cell Immunol 1986 Aug;101(1):62-71 22 UI - 86295890 AU - Glette J ; Sandberg S TI - Phototoxicity of tetracyclines as related to singlet oxygen production and uptake by polymorphonuclear leukocytes. AB - The photo-induced singlet oxygen production of six tetracyclines was measured as tryptophan degradation. Demethylchlortetracycline was the most efficient singlet oxygen producer followed by doxycycline. The least efficient producer was minocycline. Doxycycline, however, was the most potent inducer of photodamage to polymorphonuclear leukocytes (PMNLs) followed by demethylchlortetracycline. Accordingly, the singlet oxygen production during irradiation did not correlate with the induction of photodamage to the PMNLs. However, the uptake of doxycycline by the cells was 3 times higher than that of demethylchlortetracycline, and the tetracycline-induced photodamage to the PMNLs correlated with the product of singlet oxygen production during irradiation and the drug uptake by the cells. MH - Free Radicals ; Human ; In Vitro ; Neutrophils/DRUG EFFECTS/ *METABOLISM ; Oxygen/*METABOLISM ; Photosensitivity Disorders/ *CHEMICALLY INDUCED ; Support, Non-U.S. Gov't ; Tetracyclines/ *TOXICITY SO - Biochem Pharmacol 1986 Sep 1;35(17):2883-5 23 UI - 86294693 AU - Kapp A ; Kirchner H ; Wokalek H ; Sch:opf E TI - Modulation of granulocyte oxidative response by recombinant interferon alpha 2 and gamma. AB - Interferon (IFN) has been described to influence various cellular functions. In this study we investigated whether the oxidative response of polymorphonuclear leukocytes (PMN) is also affected by IFN. In order to exclude the possible influence of impurities in IFN preparations, only recombinant human IFN alpha 2 or gamma were used. Lucigenin-dependent chemiluminescence (CL) of PMN was measured to assess the production of oxygen radicals. IFN gamma at a concentration of more than 10 ng/ml elicited a minimal CL response in PMN. When PMN were incubated with IFN gamma for 1 h and then stimulated with chemotactic peptide f-met-phe (FMP), zymosan-activated serum (ZAS), zymosan particles, or phorbol-myristate acetate (PMA), the CL response was increased as consequence of the generally enhanced oxidative metabolism. IFN alpha 2 showed no such effect at any concentration tested. A 5-min pretreatment with IFN gamma decreased the ZAS response but did not affect the reaction to the other stimuli. The possibility of a generation of IFN by PMN during the assay could be excluded as no IFN activity could be detected in an antiviral assay after stimulation of PMN for 6 h with PolyI X PolyC, LPS, ConA, C. parvum, PMA, zymosan, or FMP. The modulation of granulocyte activity by IFN gamma may be important in the regulation of the anti-inflammatory response of PMN. MH - Acridines ; Free Radicals ; Granulocytes/*DRUG EFFECTS/IMMUNOLOGY/ METABOLISM ; Human ; In Vitro ; Interferon Type I/ *PHARMACODYNAMICS ; Interferon Type II/*PHARMACODYNAMICS ; Luminescence ; Oxygen/*METABOLISM ; Recombinant Proteins/ PHARMACODYNAMICS SO - Arch Dermatol Res 1986;278(4):274-6 24 UI - 86291552 AU - Yoden S ; Kida H ; Kuwabara M ; Yanagawa R ; Webster RG TI - Spin-labeling of influenza virus hemagglutinin permits analysis of the conformational change at low pH and its inhibition by antibody. AB - To study the conformational changes in the hemagglutinin (HA) molecule of A/seal/Mass/1/80 (H7N7) (Seal) influenza virus at low pH, a spin-labeling method was used. This method also permits study of antibody interaction with the HA. A synthetic nitroxide compound was used for spin-labeling of tyrosine residues of the isolated HA molecule. Electron spin resonance (ESR) spectra of the spin-labeled HA at various pH values indicated that a conformational transition occurred under acidic conditions, and around pH 5.8 the HA molecule has maximal flexibility. Since virus-induced hemolysis occurs optimally at pH 5.8-5.9, the HA molecule in the maximally flexible conformation is considered to mediate membrane fusion. The ESR spectra of the antibody-bound HA at various pH values revealed that monoclonal antibodies to different regions on the molecule may inhibit the conformational change by different modes. One antibody inhibited the changes in the HA that resulted in flexibility, while the other did not. These results support the assumption that monoclonal antibodies, which failed to inhibit hemagglutination of the virus yet neutralized viral infectivity, inhibited the fusion step in the viral replication process by interfering with a low pH-induced conformational change in the HA molecule (Kida, H., Webster, R.G. and Yanagawa, R. (1983) Arch. Virol. 76, 91-99). MH - Animal ; Antibodies, Monoclonal/*IMMUNOLOGY ; Antibodies, Viral/ *IMMUNOLOGY ; Chick Embryo ; Electron Spin Resonance ; Hemagglutinins, Viral/*ANALYSIS/IMMUNOLOGY ; Hydrogen-Ion Concentration ; Microscopy, Electron ; Orthomyxoviruses Type A/ *IMMUNOLOGY/ULTRASTRUCTURE ; Protein Conformation ; Spin Labels ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Virus Res 1986 May;4(3):251-61 25 UI - 86281462 AU - Herring FG ; Lam EW ; Burt HM TI - A spin label study of the membranolytic effects of crystalline monosodium urate monohydrate. AB - The nature of the membranolytic interaction between monosodium urate monohydrate (MSUM) crystals and phospholipid membranes was studied using electron spin resonance. Two spin probe molecules were incorporated into intact human erythrocytes and incubated with MSUM crystals. The apparent increased fluidity of 5-doxyl stearic acid incorporated erythrocytes after a 2 h incubation with MSUM was probably due to an electrostatically induced redistribution of probe from the outer more rigid layer to the fluid inner leaflet via a flip-flop mechanism. It was suggested that the MSUM induced redistribution of cationic amphiphilic probe population in the whole erythrocyte was also due to an electrostatic interaction between negatively charged MSUM crystals and positively charged probe. Possible mechanisms of MSUM induced membranolysis are discussed. MH - Crystallization ; Cyclic N-Oxides/METABOLISM ; Electron Spin Resonance ; Erythrocyte Membrane/*DRUG EFFECTS ; Erythrocytes/ METABOLISM/ULTRASTRUCTURE ; Hemolysis ; Human ; Microscopy, Electron, Scanning ; *Spin Labels ; Support, Non-U.S. Gov't ; Uric Acid/*PHARMACODYNAMICS SO - J Rheumatol 1986 Jun;13(3):623-30 26 UI - 86272312 AU - Manabe H ; Utsumi H ; Kusama T ; Hamada A TI - Micellar formation of spin-labeled fatty acyl derivatives of lipophilic muramyl dipeptides and their incorporation into liposomal membranes. AB - A lipophilic muramyl dipeptide (MDP) with a nitroxide moiety in its acyl chain (SL-MDP) and its N-methyl derivative (SL-methyl MDP) were synthesized. The SL-MDPs formed micelles (cmc, 0.1-0.3 mM). The ESR spectra of the SL-MDPs in phosphatidylcholine (PC) liposomes at 25 degrees C consisted of an anisotropic signal and three sharp lines, indicating that both SL-MDPs partitioned between membranes and aqueous phase. The amounts of the SL-MDPs in membranes depended on the phospholipid species and the cholesterol (Chol) content, but no appreciable difference was observed between SL-MDPs. The SL-MDPs partitioned well at 25 degrees C into egg yolk PC liposomes but not into pure dipalmitoylphosphatidylcholine (DPPC), suggesting that the incorporation may be related to the membrane fluidity. Chol enhanced the incorporation into both phospholipids. The mobilities of the SL-MDPs in the membranes were less than that of the corresponding spin-labeled fatty acid. Comparison of the mobilities among SL-MDPs, spin-labeled ganglioside and spin-labeled galactosylceramide showed that the hydrophilicity of the polar group may influence the immobilization of their acyl chains. MH - Acetylmuramyl-Alanyl-Isoglutamine/*ANALOGS & DERIVATIVES ; Cholesterol ; Electron Spin Resonance ; *Liposomes ; Micelles ; Models, Biological ; *Phosphatidylcholines ; Spin Labels/*CHEM SYNTHESIS ; Support, Non-U.S. Gov't SO - Chem Phys Lipids 1986 May;40(1):1-14 27 UI - 86265072 AU - Bunning RA ; Richardson HJ ; Crawford A ; Skjodt H ; Hughes D ; Evans DB ; Gowen M ; Dobson PR ; Brown BL ; Russell RG TI - The effect of interleukin-1 on connective tissue metabolism and its relevance to arthritis. AB - Interleukin-1 (IL-1) is the name given to a family of related proteins showing a variety of activities. It was originally shown to be produced by monocytes and macrophages but is now known to be produced by numerous cell types, including synovial cells. From the point of view of arthritis, its most interesting activities are those on connective tissue cells in vitro. These include stimulation of production of prostaglandins, plasminogen activator and metalloproteinases such as collagenase and proteoglycanase. IL-1 is also mitogenic for synoviocytes and bone cells, and can alter rates of production of extracellular matrix constituents. The presence of IL-1 in synovial fluids from rheumatoid and osteoarthritic joints and its actions on connective tissues in vitro suggest that IL-1 may play an important role in the pathogenesis of arthritis. There are several potential cellular sources of IL-1 in the inflamed rheumatoid joint and interactions between these cells, T lymphocytes and plasma cells may continually induce IL-1 so contributing to the chronicity of the disease. The mechanism of action of IL-1 on connective tissue cells is at present uncertain though preliminary studies suggest that IL-1 may induce cellular responses by stimulating phosphoinositide turnover and possibly protein kinase C activity. MH - Arthritis, Rheumatoid/*METABOLISM ; Cell Cycle ; Connective Tissue/*METABOLISM ; Free Radicals ; Human ; Interleukin 1/ *PHYSIOLOGY ; Osteoarthritis/METABOLISM ; Proteins/PHYSIOLOGY ; Review ; Synovial Membrane/CYTOLOGY/PHYSIOLOGY SO - Agents Actions [Suppl] 1986;18:131-52 28 UI - 86265062 AU - Masini E ; Lodovici M ; Fantozzi R ; Brunelleschi S ; Conti A ; Mannaioni PF TI - Histamine release by free radicals: paracetamol-induced histamine release from rat peritoneal mast cells after in vitro activation by monooxygenase. AB - The incubation of paracetamol with isolated rat serosal mast cells evokes the release of histamine only in the presence of (S10) liver microsomes obtained from PCB or phenobarbital treated rats. The release of histamine was not accompanied by a leakage of lactate dehydrogenase and was blocked by the antioxidant alpha-tocopherol and by D-mannitol, an hydroxyl free radicals scavenger. The data are consistent with the metabolic oxidation of paracetamol in the generation of reactive intermediates capable of activating the sequential exocytosis. MH - Acetaminophen/METABOLISM/*PHARMACODYNAMICS ; Animal ; Enzyme Induction ; Exocytosis ; Free Radicals ; Histamine Liberation/ *DRUG EFFECTS ; In Vitro ; Male ; Mast Cells/DRUG EFFECTS/ *METABOLISM ; Oxygenases/*PHARMACODYNAMICS ; Phenobarbital/ PHARMACODYNAMICS ; Rats ; Rats, Inbred Strains ; Support, Non-U.S. Gov't SO - Agents Actions 1986 Apr;18(1-2):85-8 29 UI - 86265011 AU - Fantozzi R ; Brunelleschi S ; Rubino A ; Tarli S ; Masini E ; Mannaioni PF TI - FMLP-activated neutrophils evoke histamine release from mast cells. AB - Human neutrophils, having been activated by the chemotactic peptide N-formylmethionyl-leucyl-phenylalanine (FMLP), evoke histamine release from rat serosal mast cells. The release is dependent on FMLP concentration and it can be inhibited by disodium cromoglycate and by a flavonoid, silymarin, which displays free radical scavenging properties. Silymarin inhibition of neutrophil-mediated histamine release is dose-dependent. These results further stress the concept of a neutrophil-mast cell interaction, which may be involved in inflammatory processes. MH - Adult ; Animal ; Cytochalasin B/PHARMACODYNAMICS ; Dose-Response Relationship, Drug ; Free Radicals ; Histamine Liberation/*DRUG EFFECTS ; Human ; Hydrogen Peroxide/METABOLISM ; In Vitro ; Male ; Mast Cells/*METABOLISM ; N-Formylmethionine Leucyl-Phenylalanine/*PHARMACODYNAMICS ; Neutrophils/DRUG EFFECTS/ *PHYSIOLOGY ; Rats ; Rats, Inbred Strains ; Silymarin/ PHARMACODYNAMICS ; Superoxide/METABOLISM ; Support, Non-U.S. Gov't SO - Agents Actions 1986 Apr;18(1-2):155-8 30 UI - 86258095 AU - DeLeo JA ; Floyd RA ; Carney JM TI - Increased in vitro lipid peroxidation of gerbil cerebral cortex as compared with rat. AB - The in vitro thiobarbituric acid test was used as a measure of lipid peroxidation in the gerbil and rat. Synaptosomal preparations were isolated from the cerebral cortex of each species and incubated with a free radical generating system. Varying concentrations of ADP-Fe3+, with ascorbate and oxygenated incubation medium were used to generate hydroxy-radicals. Peroxidation of the synaptosomal membrane lipids was determined using malondialdehyde (MDA) accumulation. Both the gerbil and rat demonstrated significant increases in MDA in the presence of the generating system, while the gerbil P2 fraction consistently showed an increased level of MDA accumulation as compared with rat at each of the concentrations of ADP-Fe3+. Across a range of concentrations, there was a 2-2.6-fold greater increase in MDA accumulation in gerbil as compared with rat. Free radical generation is currently thought to be involved in the associated damage following cerebral ischemia. An in vitro model capable of producing biochemically similar damage to membrane systems by means of a controlled free-radical generating system may prove useful in studying possible mechanisms of ischemic damage. MH - Animal ; Cerebral Cortex/*METABOLISM ; Cerebral Ischemia/ *METABOLISM ; Comparative Study ; Free Radicals ; Gerbillinae/ *METABOLISM ; Lipid Peroxides/*BIOSYNTHESIS ; Male ; Malondialdehyde/ANALYSIS ; Rats ; Rats, Inbred F344 ; Species Specificity SO - Neurosci Lett 1986 Jun 6;67(1):63-7 31 UI - 86255751 AU - Vapaatalo H TI - Free radicals and anti-inflammatory drugs. AB - It is widely accepted that oxygen radicals and other activated oxygen species are potent mediators or modulators of acute and chronic inflammation. They are common products of cellular metabolism, where their concentrations are controlled by different protective mechanisms such as superoxide dismutase, catalase etc. In addition to their destructive effects on various macromolecules, oxygen radicals or their products are beneficial e.g., in killing bacteria. Oxygen radicals are also closely related to arachidonic acid metabolism, prostanoids (cyclo-oxygenase pathway) and leukotrienes (lipoxygenase pathway) as well as to lipid peroxidation in general. Also, the classical mediators of inflammation, histamine and bradykinin, may be connected with the release of oxygen radicals. In addition to the earlier described inhibition of formation of prostanoids, non-steroidal anti-inflammatory drugs can inhibit production of free radicals or scavenge those already formed. Antirheumatic penicillamine and allopurinol used in the treatment of gout also act on oxygen radicals. New anti-inflammatory compounds with antioxidant properties will be developed in the near future. MH - Animal ; Anti-Inflammatory Agents/*PHARMACODYNAMICS ; Antioxidants/PHARMACODYNAMICS ; Free Radicals ; Human ; Hydrogen Peroxide/TOXICITY ; Inflammation/METABOLISM ; Oxygen/*METABOLISM ; Phagocytosis ; Prostaglandins/BIOSYNTHESIS ; Review ; Superoxide/METABOLISM ; Superoxide Dismutase/PHARMACODYNAMICS SO - Med Biol 1986;64(1):1-7 32 UI - 86242206 AU - Kusumi A ; Winkelhake JL TI - Fc:Fc interactions revealed by spin-labeled IgG heterosaccharides in model immune complexes. AB - Dynamic properties of spin-labelled heterosaccharides in the Fc-region of murine monoclonal antihapten immunoglobulin G were studied in model immune complexes (IC) as a function of the IC size. Model IC dimers, trimers and oligomers were formed using bivalent photoaffinity antigens. The ESR spectrum exhibits two components. The rotational correlation time of the less-immobilized species is shorter than 10(-10) sec, and that of the more-immobilized component is in the order to 10(-9) approximately 10(-8) sec depending on the IC size. Fraction of the more-immobilized spin labels increases, and the mobility of this component decreases with increase in IC size (i.e., mobility: monomers approximately equal to dimers greater than trimers much greater than immune-complex precipitates). These data strongly suggest the existence of Fc:Fc interactions in IC, and provide the basis for a model in which such interactions underlie the initial mechanism by which the information of antigen binding to Fab region is transferred into organized Fc:Fc association structure for IgG effector activities. MH - Animal ; Antibodies, Monoclonal ; *Antigen-Antibody Complex ; Dinitrobenzenes/IMMUNOLOGY ; Electron Spin Resonance ; Glycoproteins ; *IgG ; *Immunoglobulin Fragments ; *Immunoglobulins, Fc ; Macromolecular Systems ; Mice ; Oligosaccharides ; Spin Labels ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Biochem Biophys Res Commun 1986 May 29;137(1):237-43 33 UI - 86224197 AU - Minetti M ; Ceccarini M ; Di Stasi AM ; Petrucci TC ; Marchesi VT TI - Spectrin involvement in a 40 degrees C structural transition of the red blood cell membrane. AB - Proteins involved in a structural transition detected in red blood cell membranes at 40 degrees C by spin labeling methods have been investigated. Antibodies specific for spectrin, band 3, and protein 4.1 have been used as specific probes to modify membrane thermotropic properties. Spectrin seems to be involved in a 40 degrees C transition detected in ghosts by both a stearic acid spin label (16-doxyl stearic) and a sulfhydryl-specific maleimide analogue spin label. Circular dichroism and maleimide spin labeling studies of purified spectrin show a slow unfolding of the protein structure starting at 25-30 degrees C and a massive transition with an onset temperature of 48 and 40 degrees C, respectively. This thermotropic behavior of spectrin could be the process that modifies membrane physicochemical properties above 40 degrees C that are detected by the stearic acid spin label. The transition detected by the stearic acid spin label was modified both by antispectrin antibodies and anti-4.1 protein antibodies, but not by antibodies specific for the cytoplasmic domain of band 3. These results suggest an involvement of protein 4.1 in regulating spectrin unfolding at the membrane level. A selective inhibition of the transition detected by the maleimide spin label has been obtained with a monoclonal antispectrin antibody at 1:1 molar ratio. The involvement in this transition of a localized spectrin domain(s) containing few exposed sulfhydryl groups is proposed. MH - Antibodies/IMMUNOLOGY ; Blood Proteins/IMMUNOLOGY/METABOLISM ; Erythrocyte Membrane/*METABOLISM ; Human ; In Vitro ; Protein Conformation ; Spectrin/*METABOLISM ; Spin Labels ; Support, Non-U.S. Gov't ; Temperature ; Thermodynamics SO - J Cell Biochem 1986;30(4):361-70 34 UI - 86222602 AU - Ozaki Y ; Ohashi T ; Niwa Y TI - Oxygen radical production by neutrophils from patients with bacterial infection and rheumatoid arthritis. Measurement of hydrogen peroxide may most accurately represent enhancement of oxygen radical production during infection. AB - The production of three kinds of oxygen radicals (superoxide, hydrogen peroxide, and hydroxyl radicals) by neutrophils from patients with bacterial infection or rheumatoid arthritis was measured. The stimulators used in this study were opsonized zymosan (1 mg/ml), phorbol myristate acetate (20 ng/ml), A23187 (1 microM), and platelet activating factor (1 microM). Oxygen radical production by neutrophils from patients with rheumatoid arthritis was not significantly different from that of the control group. Hydrogen peroxide production by the neutrophils from patients with bacterial infection was significantly enhanced by only opsonized zymosan, but the production of the other kinds of oxygen radicals was not. Cytochalasin B reduced the production of hydrogen peroxide induced by opsonized zymosan more markedly than that of any other kind of oxygen radical. The measurement of hydrogen peroxide is suggested to be the most accurate indicator of the enhancement of intracellular production of oxygen radicals by neutrophils during infection. MH - A-23187/PHARMACODYNAMICS ; Arthritis, Rheumatoid/*METABOLISM ; Bacterial Infections/*METABOLISM ; C-Reactive Protein/ANALYSIS ; Comparative Study ; Cytochalasin B/PHARMACODYNAMICS ; Free Radicals ; Human ; Hydrogen Peroxide/BIOSYNTHESIS ; Leukocyte Count ; Neutrophils/DRUG EFFECTS/*METABOLISM ; Oxygen/*METABOLISM ; Platelet Activating Factor/PHARMACODYNAMICS ; Superoxide/ BIOSYNTHESIS ; Tetradecanoylphorbol Acetate/PHARMACODYNAMICS ; Zymosan/PHARMACODYNAMICS SO - Inflammation 1986 Jun;10(2):119-30 35 UI - 86222435 AU - Melinn M ; McLaughlin H TI - Hydroxyl radical scavengers inhibit human lectin-dependent cellular cytotoxicity. AB - The role of oxygen-derived free radicals (ODFR) in lectin-dependent cellular cytotoxicity (LDCC) in humans was investigated. The hydroxyl radical traps thiourea, methanol, ethanol and phenol were effective in inhibiting LDCC, as was DABCO, a singlet oxygen quencher. The proposed pathway of hydroxyl radical production in living cells is either an iron catalysed Haber-Weiss reaction or a Fenton reaction. The effect of inhibitors of these pathways was investigated. The superoxide anion scavengers superoxide dismutase, ferricytochrome c and Tiron were without effect. It was shown that Tiron inhibits the lucigenin-amplified chemiluminescence produced by the action of xanthine oxidase, and also the lucigenin-amplified chemiluminescence produced by activated PMN, suggesting that this agent (Tiron) scavenges intracellular superoxide anion. Catalase gave slight inhibition of LDCC only. The ferric iron chelator desferrioxamine gave no protection of the target cells, while the ferrous chelator, 1,10-phenanthroline, inhibited LDCC and partially prevented the detection of hydroxyl radicals generated by the Fe2+-H2O2 system. Cibacron blue, an agent that inhibits NAD(P)H linked enzymes, also inhibited LDCC. The cyclo-oxygenase inhibitors indomethacin and salicylate were without effect, while the lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA) inhibited cytolysis. None of the LDCC inhibitors was cytotoxic to the effector cells or to the target cells, neither did they inhibit lymphocyte-target binding. The findings would suggest that hydroxyl radicals have a role to play in human T-cell mediated cytolysis, either as the active lytic agent or as an epiphenomenon. MH - Antibody-Dependent Cell Cytotoxicity/*DRUG EFFECTS ; Catalase/ PHARMACODYNAMICS ; Cell Line ; Cell Survival ; Chelating Agents/ PHARMACODYNAMICS ; *Free Radicals ; Human ; Hydroxylation ; In Vitro ; Lectins/*IMMUNOLOGY ; *Oxygen ; Superoxide ; T Lymphocytes, Cytotoxic/IMMUNOLOGY SO - Immunology 1986 Jun;58(2):197-202 36 UI - 86218400 AU - Fischer SM ; Baldwin JK ; Adams LM TI - Effects of anti-promoters and strain of mouse on tumor promoter-induced oxidants in murine epidermal cells. AB - The induction of oxidant production in mouse epidermal cells by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) can be suppressed by many, but not all, known inhibitors of mouse skin tumor promotion. Members of the anti-oxidant category that were tested were ranked in the following order, 7,8-benzoflavone greater than butylated hydroxyanisole greater than ascorbic acid. In the retinoid category, retinoic acid was only moderately effective, while the trimethoxymethylphenyl analog had at least twice the inhibitory activity. Among the six protease inhibitors examined, tosylphenylalanine chloromethylketone and tosyllysine chloromethylketone were effective in diminishing the response while tosylarginine methylester, antipain, leupeptin and soybean trypsin inhibitor were ineffective, suggesting that proteases are probably not involved in the oxidant response. Several agents, trifluoperazine, trisialoganglioside and diolein, that have been shown to suppress TPA activity in other cell systems were also found to suppress the oxidant response. Finally, the extent of the oxidant response was found to correlate with sensitivity to TPA tumor promotion among the three strains of mice tested: SSIn greater than SENCAR greater than C57BL/6J. MH - Animal ; Antioxidants/*PHARMACODYNAMICS ; Diglycerides/ PHARMACODYNAMICS ; Female ; Free Radicals ; Luminescence ; Male ; Mice ; Mice, Inbred Strains ; Neutrophils/METABOLISM ; Skin/ *METABOLISM ; Skin Neoplasms/*CHEMICALLY INDUCED/PREVENTION & CONTROL ; Species Specificity ; Superoxide/METABOLISM ; Support, U.S. Gov't, P.H.S. ; Tetradecanoylphorbol Acetate/ANTAGONISTS & INHIBITORS ; Tosylarginine Methyl Ester/PHARMACODYNAMICS ; Tosyllysine Chloromethyl Ketone/PHARMACODYNAMICS ; Tosylphenylalanyl Chloromethyl Ketone/PHARMACODYNAMICS ; Trifluoperazine/PHARMACODYNAMICS SO - Carcinogenesis 1986 Jun;7(6):915-8 37 UI - 86205972 AU - Kay MM ; Bosman GJ ; Shapiro SS ; Bendich A ; Bassel PS TI - Oxidation as a possible mechanism of cellular aging: vitamin E deficiency causes premature aging and IgG binding to erythrocytes. AB - Senescent-cell antigen is a "neo-antigen: that appears on the surface of senescent cells and initiates IgG binding and cellular removal. As an approach to evaluating oxidation as a possible mechanism for generation of senescent-cell antigen, we studied erythrocytes from vitamin E-deficient rats. Vitamin E is localized primarily in cellular membranes. Its major role is the termination of free-radical chain reactions propagated by the polyunsaturated fatty acids of membrane propagated by the polyunsaturated fatty acids of membrane phospholipids. Results of our studies indicate that erythrocytes of all ages from vitamin E-deficient rats behave like old erythrocytes from normal rats, as determined by their susceptibility to phagocytosis, IgG binding, anion transport ability, and glyceraldehyde-3-phosphate dehydrogenase activity. Increased breakdown products of band 3 were observed with immunoblotting in membranes of erythrocytes from vitamin E-deficient rats. Breakdown products of band 3 are known to increase as cells age in normal individuals. The data suggest that oxidation may be a possible mechanism for erythrocyte aging and generation of senescent-cell antigen in vivo. MH - Anemia, Hemolytic/ETIOLOGY ; Animal ; Anions/BLOOD ; Band 3 Protein/PHYSIOLOGY ; Biological Transport ; *Erythrocyte Aging ; Erythrocyte Membrane/PHYSIOLOGY ; Free Radicals ; Glyceraldehydephosphate Dehydrogenase/BLOOD ; Haptoglobins/BLOOD ; Lipid Peroxides/BLOOD ; Oxidation-Reduction ; Phagocytosis ; Rats ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Vitamin E Deficiency/*BLOOD/COMPLICATIONS SO - Proc Natl Acad Sci USA 1986 Apr;83(8):2463-7 38 UI - 86188121 AU - Herrmann A ; Groth T ; Lassmann G ; Ladhoff AM ; Hillebrecht B TI - Structural alterations of the human erythrocyte membrane upon influenza virus attachment. AB - Molecular events on the human erythrocyte membrane subsequent to influence virus binding were investigated by electron spin resonance (ESR) measurements after spin labeling of the cell membrane at different positions. Virus binding affected the glycocalyx structure as well as the physical state of the cytoskeleton at the inner leaflet, but not the lipid phase. A lateral reorganization of spin-labeled glycophorin was not indicated after virus attachment. MH - Absorption ; Attachment Sites (Microbiology) ; Cell Fusion ; Comparative Study ; Cyclic N-Oxides/DIAGNOSTIC USE ; Cytoskeleton/ METABOLISM/ULTRASTRUCTURE ; Electron Spin Resonance ; Erythrocyte Membrane/*ULTRASTRUCTURE ; Glycoproteins/ANALYSIS ; Hemolysis ; Human ; Hydrogen-Ion Concentration ; Membrane Fluidity ; Microscopy, Electron ; Orthomyxoviridae/METABOLISM/*PHYSIOLOGY/ ULTRASTRUCTURE ; Polysaccharides/ANALYSIS ; Spin Labels ; Temperature SO - Biosci Rep 1986 Jan;6(1):45-55 39 UI - 86149266 AU - Subramaniam S ; Seul M ; McConnell HM TI - Lateral diffusion of specific antibodies bound to lipid monolayers on alkylated substrates. AB - We have measured the lateral mobility of fluoresceinated monoclonal IgG antibodies bound specifically to a spin label lipid hapten in phospholipid monolayers supported on alkylated silicon oxide surfaces. Dimyristoyl phosphatidylcholine and dipalmitoyl phosphatidylcholine monolayers containing 5 mol% of the lipid hapten were transferred by conventional Langmuir-Blodgett techniques onto substrates alkylated with hydrocarbon chains containing 10, 16, and 18 carbon atoms. We show that the diffusion of the bound antibodies depends on their lateral density, the composition of the lipid monolayer, and the nature of lipid coupling to hydrocarbon chains on the alkylated substrate. Antibody diffusion coefficients at low antibody densities are within a factor of 2 of those displayed by the lipid hapten in the absence of the bound antibody. High antibody densities result in reduced antibody mobility, but the lateral diffusion of unbound lipids is unaffected. MH - Antibodies ; Antigen-Antibody Complex ; Cell Membrane/IMMUNOLOGY/ *PHYSIOLOGY ; Dose-Response Relationship, Immunologic ; *Immunologic Capping ; *Membrane Fluidity ; Membrane Lipids/ *PHYSIOLOGY ; Microscopy, Fluorescence ; Models, Chemical ; Phosphatidylcholines ; Silica ; Spin Labels/IMMUNOLOGY ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. SO - Proc Natl Acad Sci USA 1986 Mar;83(5):1169-73 40 UI - 86140730 AU - Adler S ; Baker PJ ; Johnson RJ ; Ochi RF ; Pritzl P ; Couser WG TI - Complement membrane attack complex stimulates production of reactive oxygen metabolites by cultured rat mesangial cells. AB - To explore possible mechanisms by which complement membrane attack complexes (MAC) that are deposited in the glomerular mesangium might be pathogenic, we stimulated rat glomerular mesangial cells grown in vitro with nascent MACs formed from the purified human complement components C5b6 and normal human serum and measured production of superoxide ion (O2-) and hydrogen peroxide (H2O2). Mesangial cells incubated with C5b6 + serum, which results in cell membrane interaction with the MAC, produce 0.9 +/- 0.15 nmol O2-/10(5) cells per 30 min, which was significantly greater than the amount produced by cells incubated with C5b6 alone, serum alone, or decayed MACs that can no longer interact with the cell membrane (0.3 +/- 0.2, 0.4 +/- 0.1, 0.3 +/- 0.2 nmol O2-/10(5) cells per 30 min, respectively; P less than 0.02). Production of O2- after stimulation with MACs increased during the first 20 min of incubation but then plateaued. Cells exposed to decayed MACs produced small amounts of O2-, which did not increase from 20 to 60 min. Production of H2O2 was also observed after stimulation with MACs, and continued to increase during 60 min of incubation (1.22 +/- 0.16 nmol H2O2/10(5) cells per 60 min), whereas H2O2 production could not be detected after exposure to decayed MACs. Cell viability was not adversely affected by exposure to nascent MACs as determined by trypan blue exclusion or chromium-51 release. These results demonstrate that glomerular mesangial cell membrane interaction with the MAC stimulates the production of the toxic oxygen metabolites O- and H2O2. Activation of the terminal complement pathway by mesangial immune deposits in vivo might lead to tissue injury by stimulation of local production of toxic oxygen-free radicals. MH - Animal ; Cell Survival ; Cells, Cultured ; Complement/*METABOLISM ; Free Radicals ; Hydrogen Peroxide/METABOLISM ; Kidney Glomerulus/*METABOLISM ; Male ; Oxygen/*METABOLISM ; Rats ; Superoxide/METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - J Clin Invest 1986 Mar;77(3):762-7 41 UI - 86104308 AU - Sharom FJ ; Ross TE TI - Association of gangliosides with the lymphocyte plasma membrane studied using radiolabels and spin labels. AB - Gangliosides are known to act as potent suppressors of lectin-stimulated lymphocyte activation when added to the culture medium. Since this effect may be mediated via ganglioside association with (or insertion into) the plasma membrane, we have used 3H- and spin-labelled derivatives of mixed gangliosides to probe the nature of this interaction. Gangliosides bind rapidly to the lymphocyte membrane and show no preference for association with either inside-out or right-side-out membrane vesicles. Around 20% of the bound gangliosides can be removed by repetitive washing, and a further 22-28% by treatment with pronase for 1 h, suggesting that this fraction is tightly bound to membrane proteins at the cell surface. The ESR spectrum of membrane-bound gangliosides did not resemble the spin-exchanged spectrum of micellar spin-labelled gangliosides in aqueous solution, but was similar to that seen for 5 mol% ganglioside spin label in liposomes of egg phosphatidylcholine. This suggests that the bulk of the membrane-bound gangliosides are inserted and molecular dispersed in the lymphocyte membrane. Binding of wheat-germ agglutinin to lymphocyte-associated gangliosides results in specific immobilization of the carbohydrate headgroup, while concanavalin A and other lectins have little or no effect on oligosaccharide mobility. Membrane-inserted gangliosides show a response to lectin binding which is qualitatively different from that seen for gangliosides in bilayers of phosphatidylcholine. MH - Animal ; Cattle ; Cell Membrane/ANALYSIS ; Electron Spin Resonance ; Gangliosides/*ANALYSIS ; Lectins/PHARMACODYNAMICS ; Lymphocytes/*CYTOLOGY ; Mathematics ; Micelles ; Spin Labels/ *METABOLISM ; Support, Non-U.S. Gov't ; Swine SO - Biochim Biophys Acta 1986 Jan 29;854(2):287-97 42 UI - 86087670 AU - Mossmann H ; Bamberger U ; Velev BA ; Gehrung M ; Hammer DK TI - Effect of platelet-activating factor on human polymorphonuclear leukocyte enhancement of chemiluminescence and antibody-dependent cellular cytotoxicity. AB - The effect of platelet-activating factor on human granulocytes was determined by luminol-dependent chemiluminescence (CL), antibody-dependent cellular cytotoxicity (ADCC), and rosette assay. An activation of the respiratory burst could be measured by CL and be shown to depend on the presence of extracellular calcium. This direct stimulation of PMN was proved to be inhibited by oxygen radical scavengers as well as by the calcium channel blocker diltiazem. Furthermore, the presence of PAF enhanced the activation of PMN via Fc- or C3b-receptors, as demonstrated by CL and ADCC. On the other hand, no influence on the rosette-forming capacity of PMN could be detected. The results support the concept of the import role of PAF in inflammatory processes. MH - Adult ; Antibody-Dependent Cell Cytotoxicity/*DRUG EFFECTS ; Calcium/PHARMACODYNAMICS ; Free Radicals ; Human ; *Luminescence ; Neutrophils/*DRUG EFFECTS/IMMUNOLOGY/METABOLISM ; Oxygen Consumption/DRUG EFFECTS ; Platelet Activating Factor/ *PHARMACODYNAMICS ; Rosette Formation ; Stimulation, Chemical ; Support, Non-U.S. Gov't SO - J Leukocyte Biol 1986 Feb;39(2):153-65 43 UI - 86053263 AU - Kaneko M ; Leadon SA TI - Production of thymine glycols in DNA by N-hydroxy-2-naphthylamine as detected by a monoclonal antibody. AB - We have quantitated the production of thymine glycols in DNA following treatment of cultured human fibroblasts or DNA in solution with the carcinogen N-hydroxy-2-naphthylamine. Thymine glycols, detected by using a monoclonal antibody specific to this base damage, were produced in DNA in a dose dependent manner both in vitro and in vivo. Exposure of DNA to N-hydroxy-2-naphthylamine in the presence of catalase and superoxide dismutase, which break down hydrogen peroxide and superoxide anions, respectively, inhibited the production of this base damage. Thymine glycols were efficiently removed from DNA in both normal human fibroblasts and in cells from a patient with xeroderma pigmentosum complementation group A, which are deficient in nucleotide excision repair. MH - Antibodies, Monoclonal/DIAGNOSTIC USE ; Catalase/METABOLISM ; Cells, Cultured ; Dose-Response Relationship, Drug ; DNA Repair ; Free Radicals ; Glycols ; Human ; *Naphthalenes ; Superoxide Dismutase/METABOLISM ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. ; Thymine/*ANALOGS & DERIVATIVES/IMMUNOLOGY ; Xeroderma Pigmentosum/FAMILIAL & GENETIC ; *2-Naphthylamine/ ANALOGS & DERIVATIVES SO - Cancer Res 1986 Jan;46(1):71-5