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Showing 1761–1780 of 2058 publications.
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Hawkins, Clare L.; Davies, Michael J.Activated phagocyte cells generate hypochlorite (HOCl) via the release of H<inf>2</inf>O<inf>2</inf> and the enzyme myeloperoxidase. Plasma proteins are major targets for HOCl, although little information is available about the mechanism(s) of oxidation. In this study the reaction of HOCl (at least 50 ?M) with diluted fresh human plasma has been shown to generate material that oxidizes 5-thio-2-nitrobenzoic acid; these oxidants are believed to be chloramines formed from the reaction of HOCl with protein amine groups. Chloramines have also been detected with isolated plasma proteins treated with HOCl. In both cases chloramine formation accounts for approx. 20-30% of the added HOCl. These chloramines decompose in a time-dependent manner when incubated at 20 or 37C but not at 4C. Ascorbate and urate remove these chloramines in a time- and concentration-dependent manner, with the former being more efficient. The reaction of fresh diluted plasma with HOCl also gives rise to protein-derived nitrogen-centred radicals in a time- and HOCl-concentration-dependent manner; these have been detected by EPR spin trapping. Identical radicals have been detected with isolated HOCl-treated plasma proteins. Radical formation was inhibited by excess methionine, implicating protein-derived chloramines (probably from lysine side chains) as the radical source. Plasma protein fragmentation occurs in a time- and HOCl-concentration-dependent manner, as evidenced by the increased mobility of the EPR spin adducts, the detection of further radical species believed to be intermediates in protein degradation and the loss of the parent protein bands on SDS/PAGE. Fragmentation can be inhibited by methionine and other agents (ascorbate, urate, Trolox C or GSH) capable of removing chloramines and reactive radicals. These results are consistent with protein-derived chloramines, and the radicals derived from them, as contributing agents in HOCl-induced plasma protein oxidation.
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Dass, Crispin R.; Burton, Mark A.Microspheres have been used clinically for management of solid tumors. When delivered intraarterially upstream of a tumor, these spheres become entrapped in the tumor microvasculature, from where they release their drug load. Entrapment in tumor microvasculature is selective due to its aberrant characteristics compared to normal tissue vasculature. While microspheres have traditionally been utilized for targeted delivery of chemotherapeutic and radiotherapeutic agents, their versatility may be exploited for delivery of other forms of therapeutic agents. One such treatment modality is gene therapy, and the present review discusses the applicability of microspheres in delivering genes to solid tumors. The increasing usage of spheres in the literature for gene delivery attests to the rapid influx of ideas for new and radical ways of treating solid tumors.
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Kritharides, Leonard[No abstract available]
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Gao, J.; Lovejoy, David B.; Richardson, Des Raymond[No abstract available]
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Brown, Andrew J.; Jessup, Wendy K.Oxysterols are present in human atherosclerotic plaque and are suggested to play an active role in plaque development. Moreover, the oxysterol:cholesterol ratio in plaque is much higher than in normal tissues or plasma. Oxysterols in plaque are derived both non-enzymically, either from the diet and/or from in vivo oxidation, or (e.g. 27-hydroxycholesterol) are formed enzymically during cholesterol catabolism. While undergoing many of the same reactions as cholesterol, such as being esterified by cells and in plasma, certain oxysterols in some animal and in vitro models exhibit far more potent effects than cholesterol per se. In vitro, oxysterols perturb several aspects of cellular cholesterol homeostasis (including cholesterol biosynthesis, esterification, and efflux), impair vascular reactivity and are cytotoxic and/or induce apoptosis. Injection of relatively large doses of oxysterols into animals causes acute angiotoxicity whereas oxysterol-feeding experiments have yielded contrary results as far as their atherogenicity is concerned. There is no direct evidence yet in humans that oxysterols contribute to atherogenesis. However, oxysterol levels are elevated in human low-density lipoprotein (LDL) subfractions that are considered potentially atherogenic and two recent studies have indicated that raised plasma levels of a specific oxysterol (7?-hydroxycholesterol) may be associated with an increased risk of atherosclerosis. At the present time there are a number of significant and quite widespread problems with current literature which preclude more than a tentative suggestion that oxysterols have a causal role in atherogenesis. Further studies are necessary to definitively determine the role of oxysterols in atherosclerosis, and considering the wide-ranging tissue levels reported in the literature, special emphasis is needed on their accurate analysis, especially in view of the susceptibility of the parent cholesterol to artifactual oxidation.
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Irwin, Jennifer A.; tdal, Henrik; Davies, Michael J.Reaction of equine Fe(III) myoglobin with H<inf>2</inf>O<inf>2</inf> gives rise to an Fe(IV)-oxo species at the heme center and protein (globin)-derived radicals. Studies have shown that there are two (or more) sites for the protein-derived radical: at tyrosine (Tyr-103) or tryptophan (Trp-14). The latter radical reacts rapidly with oxygen to give a Trp-derived peroxyl radical. The formation of both the tyrosine phenoxyl radical and the tryptophan-derived peroxyl species have been confirmed in the present study; the latter appears to be the major initial radical, with the phenoxyl radical appearing at longer reaction times, possibly via secondary reactions. We have investigated, by EPR spectroscopy, the reactivity of the Trp-14 peroxyl radical with amino acids, peptides, proteins, and antioxidants, with the aim of determining whether this species can damage other targets, i.e., whether intermolecular protein-to-protein radical transfer and hence chain-oxidation occurs, and the factors that control these reactions. Three amino acids show significant reactivity: Tyr, Trp, and Cys, with Cys the least efficient. Evidence has also been obtained for (inefficient) hydrogen abstraction at peptide ?-carbon sites; this may result in backbone cleavage in the presence of oxygen. The myoglobin Trp-14 peroxyl radical has been shown to react rapidly with a wide range of proteins to give long-lived secondary radicals on the target protein. These reactions appear to mainly involve Tyr residues on the target protein, although evidence for reaction at Trp has also been obtained. Antioxidants (GSH, ascorbate, Trolox C, vitamin E, and urate) react with the myoglobin-derived peroxyl radical; in some cases antioxidant-derived radicals are detected. These reactions are only efficient at high antioxidant concentrations, suggesting that protein-to-protein damage transfer and protein chain-oxidation may occur readily in biological systems.
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tdal, Henrik; Andersen, Henrik Jgen; Davies, Michael J.Incubation of Fe(III)myoglobin (Fe(III)Mb) with H<inf>2</inf>O<inf>2</inf> in the presence of bovine serum albumin (BSA) has been shown previously to give albumin- derived radicals as a result of radical transfer from myoglobin to BSA. In this study the occurrence of similar processes with peroxidases has been investigated using horseradish peroxidase (HRP)/H<inf>2</inf>O<inf>2</inf>, in the presence and absence of added tyrosine. Incubation of HRP with H<inf>2</inf>O<inf>2</inf> and bovine or human serum albumins, in the presence and absence of tyrosine, gave long-lived albumin-derived radicals as detected by EPR spectroscopy. Evidence has been obtained for these albumin radicals being located on buried tyrosine residues on the basis of blocking experiments. The effect of protein conformation on radical transfer has been investigated using partial proteolytic digestion prior to protein oxidation. With HRP/H<inf>2</inf>O<inf>2</inf>/BSA and Fe(III)Mb/ H<inf>2</inf>O<inf>2</inf>/BSA increased radical concentrations were observed after limited digestion, although this effect was less marked with the HRP/H<inf>2</inf>O<inf>2</inf>/BSA system than with Fe(III)Mb/H<inf>2</inf>O<inf>2</inf>/BSA, consistent with different modes of radical transfer. More extensive digestion of BSA decreased the radical concentration to levels below those detected with native albumin, indicating that the tertiary structure of the target protein plays an important role in determining the rate of radical transfer and/or the stability of the resultant species. These results are consistent with a mechanism for the HRP/H<inf>2</inf>O<inf>2</inf>/no free tyrosine system involving radical transfer to the albumin via the heme edge of the peroxidase. In contrast, albumin radical formation by the HRP/H<inf>2</inf>O<inf>2</inf>/free tyrosine system was only marginally affected by proteolysis, consistent with free tyrosine phenoxyl radicals being the mediators of radical transfer, without significant protein-protein interaction. These protein-to-protein radical transfer reactions may have important consequences for understanding protein oxidation in biological systems.
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Lyons, Malcolm A.; Brown, Andrew J.7-Ketocholesterol is a major oxidation product of cholesterol found in human atherosclerotic plaque and is more atherogenic than cholesterol in some animal studies. 7-Ketocholesterol can inhibit cholesterol 7?-hydroxylase, the rate-limiting step in bile acid biosynthesis, as well as strongly inhibiting HMG-CoA reductase, the rate-limiting enzyme in cholesterol biosynthesis. It has even been suggested that 7-ketocholesterol is formed enzymically as an endogenous regulator of cholesterol biosynthesis. However, when tested as a pharmacological cholesterol-lowering agent, inhibition of HMG-CoA reductase was rapidly overcome and the 7-ketocholesterol metabolised. In vitro, 7-ketocholesterol has wide-ranging and potent effects, most of which have the potential to contribute to atherosclerosis. For example, 7-ketocholesterol can be cytotoxic and can induce apoptosis in vascular cells. These effects, either individually or more likely, in combination, all implicate 7-ketocholesterol in the initiation and development of atherosclerosis, but further work is needed to establish whether or not its role is a direct causal one. Copyright (C) 1999 Elsevier Science Ltd.
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Simons, Leon A.; von Konigsmark, Margaret; Simons, Judith; Stocker, Roland; Celermajer, David S.Endothelial dysfunction is thought to be an important early event in atherogenesis, related in part to reduced bioavailability of nitric oxide in the arterial wall. Endothelial function may be impaired in the presence of oxidised low density lipoprotein. The use of vitamin E as an anti-oxidant might enhance the bioavailability of nitric oxide in this situation. The effect of vitamin E 1000 IU/day on arterial endothelial physiology was studied in 20 asymptomatic older subjects, aged 45-70 years, who showed evidence of age-related endothelial dysfunction. Endothelial function was assessed non-invasively using brachial ultrasound and the primary outcome measure was flow-mediated endothelium-dependent dilatation (FMD) in response to reactive hyperaemia. A double-blind, placebo-controlled, randomised crossover design was employed. After 3 weeks of stabilisation on a standard fat-reduced diet, subjects received vitamin E or placebo for 10 weeks in random order, separated by a washout period of 8 weeks. There were no significant changes in blood pressure, plasma lipid or lipoprotein concentrations. Plasma ?-tocopherol increased from 50 3 (mean S.E.M.) to 91 6 ?mol/l (P < 0.001) with vitamin E ingestion. Total plasma F(2?)- isoprostanes, a measure of free radical-induced lipid peroxidation, were not altered by vitamin E ingestion (0.86 0.26 versus 0.82 0.25 nmol/l, P > 0.6). FMD was not significantly different between the placebo and vitamin E periods (2.7 0.6% versus 2.4 0.4%). Variation in FMD was not correlated with change in plasma ?-tocopherol (r = -0.03, P > 0.8). The study was powered to detect a minimum change in FMD of 2%. Glyceryl trinitrate endothelium-independent dilatation was not significantly changed with Vitamin E (13.7 1.3% versus 13.6 1.4%). These results exclude a major impact of medium-term supplementation with vitamin E on arterial endothelial function when age-related dysfunction is already present.
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Hazell, Linda J.; Davies, Michael J.; Stocker, RolandOxidation of low-density lipoproteins (LDL) is thought to contribute to atherogenesis. Although there is increasing evidence for a role of myeloperoxidase-derived oxidants such as hypochlorite (HOCl), the mechanism by which HOCl modifies LDL remains controversial. Some studies report the protein component to be the major site of attack, whereas others describe extensive lipid peroxidation. The present study addresses this controversy. The results obtained are consistent with the hypothesis that radical-induced oxidation of LDL's lipids by HOCl is a secondary reaction, with most HOCl consumed via rapid, non-radical reaction with apolipoprotein B-100. Subsequent incubation of HOCl-treated LDL gives rise to lipid peroxidation and antioxidant consumption in a time-dependent manner. Similarly, with myeloperoxidase/H<inf>2</inf>O<inf>2</inf>/Cl- (the source of HOCl in vivo), protein oxidation is rapid and followed by an extended period of lipid peroxidation during which further protein oxidation does not occur. The secondary lipid peroxidation process involves EPR-detectable radicals, is attenuated by a radical trap or treatment of HOCl-oxidized LDL with methionine, and occurs less rapidly when the lipoprotein was depleted of ?-tocopherol. The initial reaction of low concentrations of HOCl (400-fold or 800-fold molar excess) with LDL therefore seems to occur primarily by two-electron reactions with side-chain sites on apolipoprotein B-100. Some of the initial reaction products identified as lysine-residue-derived chloramines, subsequently undergo homolytic (one-electron) reactions to give radicals that initiate antioxidant consumption and lipid oxidation via tocopherol-mediated peroxidation. The identification of these chloramines, and the radicals derived from them, as initiating agents in LDL lipid peroxidation offers potential new targets for antioxidative therapy in atherogenesis.
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Bruce, David; Fu, Shanlin; Armstrong, Sharyn; Dean, Roger T.Oxidation of low density lipoprotein (LDL) may be atherogenic, but radical-initiated oxidation of its apoprotein B-100 (apoB) has been little studied. Transition metal ions iron and copper are candidates for mediating radical oxidation of LDL in vivo. Therefore, we studied the copper-ion-induced oxidation of apoB in human LDL. Using HPLC methods developed in our recent work, we studied the destruction of native and the generation of six oxidised amino acids; we also assessed the release of peptides from the LDL particle by FPLC. We observed time-dependent losses of apoB histidine, lysine and glycine. Long-lived reactive species, the reductant DOPA, and the oxidant hydroperoxides of valine and leucine (measured as hydroxides after reduction), were generated. Their relative abundance (mol/mol of parent amino acid) was DOPA>o- and m-tyrosine>dityrosine, valine-hydroxides, leucine hydroxides. Low molecular weight fragments were also released from the LDL in a time-dependent manner, contained hydroperoxides sensitive to GSH peroxidase, and generated radicals on reaction with iron-EDTA. The fragments contained peptides active in the quinone redox cycling procedure, comprising 0.25% of the supplied LDL amino acids. Characteristic peptides were present in each FPLC fraction containing the fragments, as judged by further HPLC fractionation. Some fragments were present in the unoxidised LDL preparations, and when these were largely removed by FPLC, copper oxidation could still generate fragments, suggesting that those present in the starting material might indicate prior oxidation. Concordantly, we found that fresh plasma LDL apoB contained ~3% of total plasma protein-bound oxidised amino acids, and with the same relative abundance. We conclude that plasma proteins including apoB are subject to physiological oxidation, similar to that inflicted by copper ions; the latter may contribute to intimal LDL oxidation, which could be the source of oxidised plasma apoB. Copyright (C) 1999 Elsevier Science Ltd.
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Stocker, RolandThe early events in atherogenesis might be due to the oxidation of low-density lipoprotein. The antioxidant vitamin E, therefore, has received much attention as a potential anti-atherogenic agent. Recent mechanistic studies of the early stage of lipoprotein-lipid oxidation show that the role of vitamin E in this process is not simply that of a classical antioxidant. Unless additional compounds are present, vitamin E can have antioxidant, neutral or pro-oxidant activity. This more complex function is reflected in the results of vitamin-E-intervention studies of atherosclerosis in animals and of controlled prospective trials on the incidence of cardiovascular disease in humans, which, overall, are inconclusive.
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Su, Tao; Cariappa, Rohit; Stanley, Keith K.In MDCK cells, N-glycans have been shown to determine the sorting of secretory proteins and membrane proteins to the apical domain in the absence of a dominant basolateral targeting signal. We have examined the sorting of endogenous proteins in ECV304 cells in the presence and absence of tunicamycin, an inhibitor of N-linked glycosylation. A prominent apically secreted protein of 71 kDa was not N-glycosylated and continued to be secreted apically in the presence of tunicamycin. In contrast, other endogenous proteins that were N-glycosylated were secreted preferentially into the basolateral medium or without polarity. When rat growth hormone was expressed in MDCK and ECV304 cells, we observed 65 and 94% of the secretion to the basolateral medium, respectively. Introduction of a single N-glycan caused 83% of the growth hormone to be secreted at the apical surface in MDCK cells but had no significant effect on the polarity of secretion of growth hormone in ECV304 cells. These results indicate that not all cell lines recognise N-glycans as a signal for apical sorting and raises the possibility of using ECV304 cells as a model system for analysis of apical sorting molecules.
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Upston, Joanne M.; Karjalainen, Ari; Bygrave, Fyfe L.; Stocker, RolandAscorbate (AH, the reduced form of vitamin C) is an important radical scavenger and antioxidant in human plasma; the resulting ascorbyl radical can disproportionate to AH and dehydroascorbic acid (DHA). Here we address potential maintenance mechanism(s) for extracellular AH by examining the ability of cells to convert extracellularly presented DHA to AH. DHA was rapidly transported into human liver (HepG2), endothelial and whole blood cells in vitro by plasma membrane glucose transporters and reduced intracellularly. Liver cells displayed the highest capacity to release the intracellularly accumulated AH. The proteins responsible for DHA uptake and AH release could be distinguished by inhibitor studies. Thus, unlike DHA uptake, AH efflux was largely insensitive to cytochalasin B and thiol-reactive agents but was inhibited by phloretin, 4,4'-di-iso-thiocyanostilbene-2,2'-disulphonate and isoascorbate. Efflux of AH from cells was temperature-sensitive and saturable with a low affinity (millimolar, intracellular) for AH. In addition to isolated liver cells, perfusion of intact rat and guinea-pig liver with DHA resulted in AH in the circulating perfusate. Our results show that hepatocytes take up and reduce DHA and subsequently release part of the AH formed, probably via a membrane transporter. By converting extracellular DHA to extracellular AH, the liver might contribute to the maintenance of plasma AH, a process that could be important under conditions of oxidative stress.
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Mohr, Detlef; Umeda, Yoh; Redgrave, Trevor G.; Stocker, RolandDietary oxysterols can reach the circulation and this may contribute to atherosclerosis, where lipid oxidation is thought to be important. There is also evidence that, in rats, peroxidized lipids are absorbed and transported into lymph, although the method used to detect lipid peroxides lacked specificity. We tested whether intragastric administration of vegetable oils containing triglyceride hydroperoxides (TG-OOH) to rats resulted in detectable lipid hydroperoxides in mesenteric lymph. Using sensitive HPLC with postcolumn chemiluminescence detection, we were unable to detect hydroperoxides of triglycerides, cholesterylesters or phospholipids during the course of lipid absorption, and lymph levels of ascorbate, urate, ?-tocopherol and ubiquinol-9 did not change significantly. By contrast, we observed a striking reducing activity judged by the efficient reduction of administered ubiquinones-9 and -10 to the corresponding ubiquinols. Exposure of rat lymph and isolated chylomicrons to aqueous peroxyl radicals revealed patterns of antioxidant consumption and lipid hydroperoxide formation similar to those described previously for human extravascular fluids and isolated lipoproteins, respectively. In particular, rates of TG-OOH formation in lymph and chylomicrons were very low to undetectable as long as ascorbate and/or ubiquinols were present, but subsequently proceeded in a chain reaction despite the presence of ?-tocopherol. These studies demonstrate that rat intestine and mesenteric lymph possess efficient antioxidant defenses against preformed lipid hydroperoxides and (peroxyl) radical mediated lipid oxidation. We conclude that dietary lipid hydroperoxides or postprandial oxidation of lipids are not likely to contribute to these particular forms of oxidized lipids in circulation and aortic tissue.
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Rees, David; Sloane, Timothy M.; Jessup, Wendy K.; Dean, Roger T.; Kritharides, LeonardApolipoprotein A-I (apoA-I) overexpression inhibits atherogenesis in mice, and apolipoprotein E (apoE) secreted by foam cell macrophages may exert antiatherogenic effects within the arterial wall. We hypothesized that interaction between apoA-I and apoE contributed to the antiatherogenic properties of apoA-I, and therefore investigated whether apoA-I stimulated secretion of apoE by foam cell macrophages. Cholesterol enrichment of primary murine and human macrophages increased spontaneous apoE secretion 2-fold, as quantified by Western blot and chemiluminescence detection. Human apoA-I caused a further marked increase of apoE secretion from both murine (3.8- fold, p < 0.01) and human (3.2-fold, p = 0.01) foam cells in a time- and concentration- dependent manner, and this increase was confirmed by immunoprecipitation of [35S]methionine-labeled macrophage apoE. The protein synthesis inhibitor cycloheximide, but not the transcription inhibitor actinomycin D, markedly inhibited apoE secretion to apoA-I (73.1 9.8% inhibition at 4 h) and completely suppressed apoE secretion beyond 4 h. Pretreatment of macrophages with Pronase inhibited initial apoA-I-mediated apoE secretion by 70.5 6.5% at 2 h, but by 8 h apoA-I-induced apoE secretion was the same in Pronasepretreated and non-pretreated cells. Non- apolipoprotein-mediated cholesterol efflux induced by trimethyl-? cyclodextrin did not enhance apoE secretion, whereas phospholipid vesicles inducing the same degree of cholesterol efflux substantially enhanced apoE secretion, and apoA-I and phospholipid vesicles in combination demonstrated additive induction of apoE secretion. We conclude that apoA-I concurrently stimulates apoE secretion and cholesterol efflux from foam cell macrophages and that lipoprotein-derived apoA-I may enhance local secretion and accumulation of apoE in atherosclerotic lesions.
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Adams, Mark R.; Phu, Co Vien; Stocker, Roland; Celermajer, David S.L-Arginine, the physiologic substrate of nitric oxide synthase, has antiatherogenic properties in animal models of atherosclerosis, and improves endothelial function in hypercholesterolemic humans. Some of these effects may be mediated by increased production of nitric oxide; however, some investigators have postulated a direct antioxidant action related to its aminoguanidine moiety. We aimed therefore, to assess the antioxidant properties of L-arginine. The antioxidant properties of 200 ?M L-arginine, 200 ?M D-arginine and 200 ?M L-glutamate were compared with the powerful antioxidant ascorbate and a control (phosphate-buffered saline). Compounds were tested using four in vitro methods: (1) the Esterbauer technique (which tests the ability of the compounds to scavenge free radicals or chelate transition metals); (2) the effect on the autoxidation of ascorbate; (3) anti-tocopherol mediated peroxidation (which tests the compound's ability to synergize with ?-tocopherol to prevent mild chemically induced LDL oxidation); and (4) the ability of the compounds to attenuate ?-tocopherol radical in micellular emulsions (TRAA). The above methods were repeated using the metabolites of the test compounds after incubation with human endothelial cells. Ex vivo studies were then carried out by measuring levels of lipid peroxide production (using HPLC with UV and chemiluminescence detection) in three healthy volunteers before and 2 h after a single 7-g oral dose of L-arginine. By the Esterbauer technique, L-arginine increased lag time by 45% compared to control, as did D-arginine by 50%; L-glutamate had no effect and ascorbate increased lag time by 325%. Neither L-arginine, D-arginine or L-glutamate had significant effects on the autoxidation of ascorbate or anti-tocopherol mediated peroxidation. By the TRAA method, L-arginine had a small effect on preventing the decay of tocopherol. The results were similar for the studies of the compound's metabolites. In ex vivo studies, no changes were seen in lipid peroxide levels following acute dosage with L-arginine. L-Arginine has only weak and non-specific antioxidant effects, suggesting that its major cardioprotective benefits occur through other mechanisms, such as via the nitric oxide pathway. Copyright (C) 1999 Elsevier Science Ireland Ltd.
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Tomasetti, Marco; Littarru, Gianpaolo; Stocker, Roland; Alleva, RenataUbiquinol-10, the reduced form of coenzyme Q<inf>10</inf>, is a powerful antioxidant in plasma and lipoproteins. It has been suggested that endogenous ubiquinol-10 also exerts a protective role even towards DNA oxidation mediated by lipid peroxidation. Even though the antioxidant activity of coenzyme Q<inf>10</inf> is mainly ascribed to ubiquinol-10, a role for ubiquinone-10 (the oxidized form), has been suggested not only if appropriate reducing systems are present. To investigate whether the concentration of ubiquinol-10 or ubiquinone-10 affects the extent of DNA damage induced by H<inf>2</inf>O<inf>2</inf>, we supplemented in vitro human lymphocytes with both forms of coenzyme Q<inf>10</inf> and evaluated the DNA strand breaks by Comet assay. The exposure of lymphocytes to100 ?M H<inf>2</inf>O<inf>2</inf> resulted in rapid decrease of cellular ubiquinol-10 content both in ubiquinol-10-enriched and in control cells, whereas ?-tocopherol and ?-carotene concentration were unchanged. After 30 min from H<inf>2</inf>O<inf>2</inf> exposure, the amount of DNA strand breaks was lower and cells' viability was significantly higher in ubiquinol-10-enriched cells compared with control cells. A similar trend was observed in ubiquinone-10-enriched lymphocytes when compared with control cells. Our experiments suggest that coenzyme Q<inf>10</inf> in vitro supplementation enhances DNA resistance towards H<inf>2</inf>O<inf>2</inf>-induced oxidation, but it doesn't inhibit directly DNA strand break formation. Copyright (C) 1999 Elsevier Science Inc.
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Luxford, Catherine; Morin, Bicte; Dean, Roger T.; Davies, Michael J.Exposure of amino acids, peptides and proteins to radicals, in the presence of oxygen, gives high yields of hydroperoxides. These materials are readily decomposed by transition metal ions to give further radicals. We hypothesized that hydroperoxide formation on nuclear proteins, and subsequent decomposition of these hydroperoxides to radicals, might result in oxidative damage to associated DNA, We demonstrate here that exposure of histone H1 and model compounds to ?-radiation in the presence of oxygen gives hydroperoxides in a dose-dependent manner. These hydroperoxides decompose to oxygen- and carbon-centred radicals (detected by electron paramagnetic resonance spectroscopy) on exposure to Cu+ and other transition metal ions. These hydroperoxide-derived radicals react readily with pyrimidine DNA bases and nucleosides to give adduct species (i.e. protein-DNA base cross-links). Product analysis has demonstrated that radicals from histone H1-hydroperoxides, and other protein and amino acid hydroperoxides, can also oxidize both free 2'-deoxyguanosine and intact calf thymus DNA to give the mutagenic oxidized base 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-hydroxy-2'-deoxyguanosine, 8-oxodG). The yield of 7,8-dihydro-8-oxo-2'-deoxyguanosine is proportional to the initial protein-hydroperoxide concentration, and corresponds (for histone H1-hydroperoxide. 280 ?M) to approx. 1.4% conversion for free 2'-deoxyguanosine (200 ?M), and 0.14% for 2'-deoxyguanosine in DNA (70 ?g/ml). Evidence has also been obtained with DNA for reaction at cytosine and thymine, but not adenine; the lack of damage to the latter may result from damage transfer to 2'-deoxyguanosine residues. These studies demonstrate that initial radical-induced damage to nuclear proteins can give rise to subsequent DNA damage, the latter includes both DNA-protein cross-links and formation of oxidized DNA bases.
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Adams, Mark R.; Celermajer, David S.Atherosclerosis, the pathological process underlying myocardial infarction, stroke and other occlusive vascular disease, is the major cause of death in the Western world. The development of techniques to accurately and reproducibly detect and measure the early changes of atherosclerosis and/or to identify subjects at highest cardiovascular risk may aid in the development of prevention strategies and facilitate a decrease in morbidity and mortality from atherosclerosis. Increasing understanding of the pathophysiology of early atherosclerosis has allowed the development of a number of potential methods for the assessment of the early stages of atherosclerosis in humans. These include techniques for assessing early structural changes in the coronary arteries with electron-beam computed tomography and magnetic resonance imaging. External vascular ultrasound has also been used to image other circulations as a surrogate marker for coronary acherosclerosis, e.g. the measurement of carotid artery intimamedia thickness. Early functional changes of atherosclerosis have also been described many years before the development of structural changes. A number of techniques have been developed to measure endothelial dysfunction, one of the earliest changes of atherosclerosis, including noninvasive measurement of endothelial function using external vascular ultrasound. A variety of serum markers have also been described, and may be useful markers of atherosclerosis. We discuss some of the more promising techniques for the detection of early, presymptomatic atherosclerosis.
