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Radical Generation (radical + generation)
Kinds of Radical Generation Selected AbstractsFree radical generation and oxidative stress with ageing and exercise: Differential effects in the myocardium and liverACTA PHYSIOLOGICA, Issue 4 2000Bejma Reactive oxygen species and other oxidants are implicated in the mechanisms of biological ageing and exercise-induced tissue damage. The present study examined the effects of ageing and an acute bout of exercise on intracellular oxidant generation, lipid peroxidation, protein oxidation and glutathione (GSH) status in the heart and liver of young adult (8 month, N=24) and old (24 month, N=24) male Fischer 344 rats. Young rats ran on treadmill at 25 m min,1, 5% grade until exhaustion (55.4 ± 2.7 min), whereas old rats ran at 15 m min,1, 5% until exhaustion (58.0 ± 2.7 min). Rate of dichlorofluorescin (DCFH) oxidation, an indication of intracellular oxidant production, was significantly higher in the homogenates of aged heart and liver compared with their young counterparts. In the isolated heart and liver mitochondria, ageing increased oxidant production by 29 and 32% (P < 0.05), respectively. Acute exercise increased oxidant production in the aged heart but not in the liver. When nicodinamide dinucleotide phosphate (reduced), adenosine diphosphate and Fe3+ were included in the assay, DCFH oxidation rate was 47 and 34% higher (P < 0.05) in the aged heart and liver homogenates, respectively, than the young ones. The age differences in the induced state reached 83 and 140% (P < 0.01) in isolated heart and liver mitochondria, respectively. Lipid peroxidation was increased in the aged liver and exercised aged heart, whereas protein carbonyl content was elevated only in the aged heart (P < 0.05). Although our data using DCFH method probably underestimated cellular oxidant production because of time delay and antioxidant competition, it is clear that oxidative stress was enhanced in both heart and liver with old age. Furthermore, aged myocardium showed greater susceptibility to oxidative stress after heavy exercise. [source] High glucose activates pituitary proopiomelanocortin gene expression: possible role of free radical-sensitive transcription factorsDIABETES/METABOLISM: RESEARCH AND REVIEWS, Issue 4 2007Koichi Asaba Abstract Background Hyperglycemia is recognized as a metabolic stress, and indeed it is known to stimulate hypothalamo-pituitary-adrenal (HPA) axis, a representative anti-stress system, in patients with diabetes mellitus or in animal models of hyperglycemia. Thus, we tried to clarify the molecular mechanism of glucose-induced HPA axis activation. Methods We studied the effect of high glucose on the transcriptional regulation of proopiomelanocortin (POMC) gene that encodes adrenocorticotropic hormone, a central mediator of HPA axis, using AtT20 corticotroph cell line in vitro. Results We found that high glucose concentration (24 mM) significantly stimulated the 5,-promoter activity of POMC gene. The effect was promoter-specific, and was mimicked by nuclear factor-kappaB (NF-,B)- or AP1-responsive promoters but not by cAMP-responsive element or serum-response element-containing promoters. Furthermore, the stimulatory effect of high glucose on POMC gene was eliminated by NF-,B and AP1 inhibitors, suggesting the involvement of the transcriptional factors. The POMC 5,-promoter has the canonical NF-,B consensus sequence, and gel shift assay showed the binding of NF-,B to the element. Finally, the effect of high glucose was completely abolished by treatment with a radical quencher 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL). Conclusions Our data suggest that hyperglycemia activates POMC gene expression, at least partly, via NF-,B/AP1, and that high-glucose-induced free radical generation may mediate the activation of these transcription factors, which in turn stimulates the transcription of POMC gene. Copyright © 2006 John Wiley & Sons, Ltd. [source] Immobilized Cytochrome c Sensor in Organic/Aqueous Media for the Characterization of Hydrophilic and Hydrophobic AntioxidantsELECTROANALYSIS, Issue 18 2003Moritz Beissenhirtz Abstract A method for the characterization of antioxidants is introduced, which allows the measurement of pure hydrophilic and hydrophobic substances as well as complex cosmetic creams. The sensor is based on cytochrome c covalently immobilized on a gold wire electrode working in mixtures of phosphate buffer and organic solvents. It is combined with a superoxide generating enzyme system. The decrease of the superoxide concentration in the test solution by the added antioxidants is detected and used for the quantification of their antioxidative efficiency. Electrochemical properties of immobilized cytochrome c, such as formal potential and heterogeneous electron transfer rate constant, have been investigated in mixtures of aqueous buffer and DMSO, methanol, butanediol, and THF. The maximum organic solvent content for quasi-reversible electrode behavior was correlated to spectroscopic measurements. The activity of the radical producing enzyme in such media was determined and the radical generation characterized. The antioxidative properties of pure substance such as ascorbic acid and Biochanin A as well as of five anti-ageing cosmetic creams were studied. This showed also the influence of matrix composition on the efficiency of antioxidative supplements. [source] The alpha-amino group of l -arginine mediates its antioxidant effectEUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 2 2001S. Wallner Antioxidant effects may constitute part of the possible antiatherogenic effects of the amino acid l -arginine. These antioxidant properties were further characterized in a model of lipoprotein oxidation. Oxidation of lipoproteins in unfractionated human serum was continuously monitored by a fluorescent probe. The antioxidant effects of l -arginine, N -,-acetyl-arginine and vitamin E in combination with l -arginine were measured after initiation of free radical generation with either copper or 2,2,-azobis(2-amidinopropane) hydrochloride (AAPH). The half-time of the fast propagation rate for copper-induced lipoprotein oxidation increased after incubation with l -arginine in a dose-dependent manner (P < 0·01). N -,-acetyl-arginine did not show such effects. Vitamin E and l -arginine show different effects on copper-induced oxidation, the former increasing only lag-time, the latter increasing only propagation rate, and do not have reciprocal effects. In contrast to copper-induced oxidation, l -arginine increased the lag-time of AAPH-induced lipoprotein oxidation (P < 0·01), with no effect on the propagation rate at physiological concentrations. Again, N -,-acetyl-arginine did not show any antioxidant effects. Our experiments provide further evidence that mechanisms other than serving as a substrate for the NO-synthase could be involved in the antiatherosclerotic effect of l -arginine. In addition, our experiments clearly show, that the antioxidant effect of l -arginine is due to a chemical moiety different from that serving as the substrate for NO biosynthesis. [source] Selective detection of superoxide anion radicals generated from macrophages by using a novel fluorescent probeFEBS JOURNAL, Issue 7 2007Jing Jing Gao Quantitation of superoxide radical (O,2,·) production at the site of radical generation remains challenging. A simple method to detect nanomolar to micromolar levels of superoxide radical in aqueous solution has been developed and optimized. This method is based on the efficient trapping of O2,· using a novel fluorescent probe (2-chloro-1,3-dibenzothiazolinecyclohexene), coupled with a spectra character-signaling increase event. A high-specificity and high-sensitivity fluorescent probe was synthesized in-house and used to image O2,· in living cells. Better selectivity for O2,· over competing cellular reactive oxygen species and some biological compounds illustrates the advantages of our method. Under optimal conditions, the linear calibration range for superoxide anion radicals was 5.03 × 10,9,3.33 × 10,6 m. The detection limit was 1.68 × 10,9 m. Fluorescence images of probe-stained macrophages stimulated with 4,-phorbol 12-myristate 13-acetate were obtained successfully using a confocal laser scanning microscope. [source] Superoxide radical generation and Mn- and Cu-Zn superoxide dismutases activities in human leukemic cellsHEMATOLOGICAL ONCOLOGY, Issue 1 2003Masahiko Kato Abstract Mn- and Cu-Zn superoxide dismutase (SOD) activities and generation of superoxide radicals (O) were assessed in leukemic cells from 10 patients with acute myeloid or monocytic leukemia (AML) and 10 patients with acute lymphoblastic leukemia (ALL), using a sensitive, specific chemiluminescence method. Leukemic cells were classified according to the French,American,British classification. M4 AML cells from two patients produced some O upon stimulation with opsonized zymosan (OZ), phorbol myristate acetate (PMA) or N-formyl-methionyl-leucyl-phenylalanine (FMLP), but less than normal granulocytes or monocytes. M5b AML cells from one patient produced as much O in response to these stimulants as normal monocytes. No O generation was induced in other types of leukemic cells. Total SOD activity in AML cells was significantly greater in normal granulocytes, but was only half of the activity in ALL cells. Mn-SOD in AML cells was very low or undetectable. These results suggest that except in M5b cells, decreased O production may contribute to susceptibility to infections in AML patients. Decreased Mn-SOD activity in AML cells may predispose them to oxidative stress. Copyright © 2002 John Wiley & Sons, Ltd. [source] Less-oxidative hemodialysis solution rendered by cathode-side application of electrolyzed waterHEMODIALYSIS INTERNATIONAL, Issue 3 2007Masaaki NAKAYAMA Abstract Electrolyzed water (EW) generated on the cathode side reportedly displays anti-oxidative properties, and application of EW to hemodialysis (HD) systems supposedly suppresses oxidative markers in patients on HD. However, most of the chemical properties and biological effects of such solutions remain unclear. This study aimed to examine those issues to clarify the scientific background for the clinical use of EW solution. Reverse osmosis water comprising EW from the cathode side (e-RO) was prepared and used to process a test HD solution (e-HD). Chemical and biological properties of these solutions were compared with controls. Redox properties were examined by chemiluminescence (CL) of the luminol-H2O2 system. Biological effects of e-RO on human polymorphonuclear leukocytes (PMNs) were tested with respect to the cellular protection against methylglyoxal, and with respect to the preservation of cellular function as to radical generation. Control HD solution presented the highest CL, followed by e-HD, control RO, suggesting a lower oxidative capacity for EW-based solutions. Increased levels of dissolved hydrogen were characteristic of e-RO and e-HD. Application of e-RO tended to be associated with less injury of PMNs by methylglyoxal, and with significantly higher levels of radical generation compared with the control. Compared with control HD, e-RO-based HD solution displays less-oxidative capacity in chemical terms, and may at least partly facilitate preservation of PMN viability. These results appear to offer a scientific basis for supporting the clinical challenge of applying this technology to HD treatment. [source] Bid-dependent generation of oxygen radicals promotes death receptor activation,induced apoptosis in murine hepatocytesHEPATOLOGY, Issue 2 2004Wen-Xing Ding Activation of tumor necrosis factor receptor 1 or Fas leads to the generation of reactive oxygen species, which are important to the cytotoxic effects of tumor necrosis factor , (TNF-,) or Fas ligand. However, how these radicals are generated following receptor ligation is not clear. Using primary hepatocytes, we found that TNF-, or anti,Fas antibody,induced burst of oxygen radicals was mainly derived from the mitochondria. We discovered that Bid,a pro-death Bcl-2 family protein activated by ligated death receptors,was the main intracellular molecule signaling the generation of the radicals by targeting to the mitochondria and that the majority of oxygen radical production was dependent on Bid. Reactive oxygen species contributed to cell death and caspase activation by promoting FLICE-inhibitory protein degradation and mitochondrial release of cytochrome c. For the latter part, the oxygen radicals did not affect Bak oligomerization but instead promoted mitochondrial cristae reorganization and membrane lipid peroxidation. Antioxidants could reverse these changes and therefore protect against TNF-, or anti,Fas-induced apoptosis. In conclusion, our studies established the signaling pathway from death receptor engagement to oxygen radical generation and determined the mechanism by which reactive oxygen species contributed to hepatocyte apoptosis following death receptor activation. (HEPATOLOGY 2004;40:403,413.) [source] Effects of nickel poisoning on expression pattern of the 72/73 and 94 kDa stress proteins in rat organs and in the COS-7, HepG2, and A549 cell linesJOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, Issue 1 2005N. Hfaiedh Abstract The present study deals with the effects of Ni on the expression level of three stress proteins, namely, the cytosolic HSP72 and HSP73, and the reticulum-associated GRP94. Experiments were carried out on "Wistar" female rats daily injected with 4 mg NiCl2 per kg body weight for 1, 3, 5, and 10 days. Another set of experiments were carried out using cell lines, derived from the monkey kidney (COS-7), and from human tumors of the lung (A549) and liver (HepG2). Cells were cultured for 4 days in the permanent presence of 100, 200, or 400 µM NiCl2. In control rats, stress proteins pattern was found to be tissue specific: two protein bands of 96 and 94 kDa were immunodetected with the anti-GRP94 antibody in kidney and liver extracts, whereas only the 96 kDa band was present in ovary extracts. HSP73 was present in kidney, liver, and ovary whereas HSP72 was only found in kidney. In kidney of nickel-treated animals, HSP73 and the 96 kDa proteins were overexpressed whereas HSP72 was strongly down regulated. No such effect was observed in liver or ovary. Similarly, in nickel-treated cell lines, HSP72 was downregulated and GRP94 (96 kDa protein) was overexpressed. HSP73 expression appeared moderately increased in A549 cells but decreased in COS-7 cells. Because long-term caloric restriction was reported to reduce free radical generation in cells, the effect of 1 month food restriction (50%) was tested in rats as a possible way to lower oxidative damages induced by Ni. No significant effect on HSP expression was observed. © 2005 Wiley Periodicals, Inc. J Biochem Mol Toxicol 19:12,18, 2005; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20056 [source] Synergistic dopaminergic neurotoxicity of manganese and lipopolysaccharide: differential involvement of microglia and astrogliaJOURNAL OF NEUROCHEMISTRY, Issue 2 2010Ping Zhang Abstract Overexposure to manganese is known to cause damage to basal ganglial neurons and the development of movement abnormalities. Activation of microglia and astrocytes has increasingly been associated with the pathogenesis of a variety of neurological disorders. We have recently shown that microglial activation facilitates manganese chloride (MnCl2, 10,300 ,M)-induced preferential degeneration of dopamine (DA) neurons. In this study, we report that combinations of MnCl2 (1,30 ,M) and endotoxin lipopolysaccharide (LPS, 0.5,2 ng/mL), at minimally effective concentrations when used alone, induced synergistic and preferential damage to DA neurons in rat primary neuron-glia cultures. Mechanistically, MnCl2 significantly potentiated LPS-induced release of tumor necrosis factor-alpha and interleukin-1 beta in microglia, but not in astroglia. MnCl2 and LPS were more effective in inducing the formation of reactive oxygen species and nitric oxide in microglia than in astroglia. Furthermore, MnCl2 and LPS-induced free radical generation, cytokine release, and DA neurotoxicity was significantly attenuated by pre-treatment with potential anti-inflammatory agents minocycline and naloxone. These results demonstrate that the combination of manganese overexposure and neuroinflammation is preferentially deleterious to DA neurons. Moreover, these findings not only shed light on the understanding of manganese neurotoxicity but may also bear relevance to the potentially multifactorial etiology of Parkinson's disease. [source] Ca2+ -induced permeabilization promotes free radical release from rat brain mitochondria with partially inhibited complex IJOURNAL OF NEUROCHEMISTRY, Issue 3 2005Tatyana V. Votyakova Abstract Mitochondrial complex I dysfunction has been implicated in a number of brain pathologies, putatively owing to an increased rate of reactive oxygen species (ROS) release. However, the mechanisms regulating the ROS burden are poorly understood. In this study we investigated the effect of Ca2+ loads on ROS release from rat brain mitochondria with complex I partially inhibited by rotenone. The addition of 20 nm rotenone to brain mitochondria increased ROS release. Ca2+ (100 µm) alone had no effect on ROS release, but greatly potentiated the effects of rotenone. The effect of Ca2+ was decreased by ruthenium red. Ca2+ -challenged mitochondria lose about 88% of their glutathione and 46% of their cytochrome c under these conditions, although this depends only on Ca2+ loading and not complex I inhibition. ADP in combination with oligomycin decreased the loss of glutathione and cytochrome c and free radical generation. Cyclosporin A alone was ineffective in preventing these effects, but augmented the protection provided by ADP and oligomycin. Non-specific permeabilization of mitochondria with alamethicin also increased the ROS signal, but only when combined with partial inhibition of complex I. These results demonstrate that Ca2+ can greatly increase ROS release by brain mitochondria when complex I is impaired. [source] Epidermal Growth Factor Induces Oxidative Neuronal Injury in Cortical CultureJOURNAL OF NEUROCHEMISTRY, Issue 1 2000Yoo Kyung Cha Abstract : Recently, we have demonstrated that certain neurotrophic factors can induce oxidative neuronal necrosis by acting at the cognate tyrosine kinase-linked receptors. Epidermal growth factor (EGF) has neurotrophic effects via the tyrosine kinase-linked EGF receptor (EGFR), but its neurotoxic potential has not been studied. Here, we examined this possibility in mouse cortical culture. Exposure of cortical cultures to 1-100 ng/ml EGF induced gradually developing neuronal death, which was complete in 48-72 h ; no injury to astrocytes was noted. Electron microscopic findings of EGF-induced neuronal death were consistent with necrosis ; severe mitochondrial swelling and disruption of cytoplasmic membrane occurred, whereas nuclei appeared relatively intact. The EGF-induced neuronal death was accompanied by increased free radical generation and blocked by the anti-oxidant Trolox. Suggesting mediation by the EGFR, an EGFR tyrosine kinase-specific inhibitor, C56, attenuated EGF-induced neuronal death. In addition, inhibitors of extracellular signal-regulated protein kinase 1/2 (Erk-1/2) (PD98056), protein kinase A (H89), and protein kinase C (GF109203X) blocked EGF-induced neuronal death. A p38 mitogen-activated protein kinase inhibitor (SB203580) or glutamate antagonists (MK-801 and 6-cyano-7-nitroquinoxaline-2,3-dione) showed no protective effect. The present results suggest that prolonged activation of the EGFR may trigger oxidative neuronal injury in central neurons. [source] Astaxanthin, a dietary carotenoid, protects retinal cells against oxidative stress in-vitro and in mice in-vivoJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 10 2008Yoshimi Nakajima We have investigated whether astaxanthin exerted neuroprotective effects in retinal ganglion cells in-vitro and in-vivo. In-vitro, retinal damage was induced by 24-h hydrogen peroxide (H2O2) exposure or serum deprivation, and cell viability was measured using a WST assay. In cultured retinal ganglion cells (RGC-5, a rat ganglion cell-line transformed using E1A virus), astaxanthin inhibited the neurotoxicity induced by H2O2 or serum deprivation, and reduced the intracellular oxidation induced by various reactive oxygen species (ROS). Furthermore, astaxanthin decreased the radical generation induced by serum deprivation in RGC-5. In mice in-vivo, astaxanthin (100 mg kg,1, p.o., four times) reduced the retinal damage (a decrease in retinal ganglion cells and in thickness of inner plexiform layer) induced by intravitreal N -methyl- d -aspartate (NMDA) injection. Furthermore, astaxanthin reduced the expressions of 4-hydroxy-2-nonenal (4-HNE)-modified protein (indicator of lipid peroxidation) and 8-hydroxy-deoxyguanosine (8-OHdG; indicator of oxidative DNA damage). These findings indicated that astaxanthin had neuroprotective effects against retinal damage in-vitro and in-vivo, and that its protective effects may have been partly mediated via its antioxidant effects. [source] Cimetidine: antioxidant and metal-binding propertiesJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 12 2002Zaynab Lambat ABSTRACT Cimetidine is one of the most potent H2 receptor antagonists for inhibiting excessive histamine-induced acid secretion and is currently used worldwide to treat peptic ulcers. In this study, levels of free radicals were assessed and the ability of cimetidine to act as an antioxidant was determined using nitroblue-tetrazolium assay and lipid peroxidation assays. Free radical generation in the brain is promoted by the presence of iron, as occurs in the Fenton reaction. The results show that cimetidine reduces the generation of superoxide anion formed in the nitroblue-tetrazolium assay. In addition, cimetidine (1 mm) is able to reduce the iron-induced rise in lipid peroxidation in rat brain homogenates. Electrochemistry, UV/Vis spectroscopy and HPLC experiments show metal-ligand interactions between cimetidine and transition metals. The results imply that cimetidine provides a neuroprotective effect by binding to iron and copper, thus making them unavailable for free radical production. [source] Incubation period in the 2,2,4,4-tetramethyl-1-piperidinyloxy-mediated thermal autopolymerization of styrene: Kinetics and simulationsJOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 24 2006Enrique Saldķvar-Guerra Abstract Mechanisms and simulations of the induction period and the initial polymerization stages in the nitroxide-mediated autopolymerization of styrene are discussed. At 120,125 °C and moderate 2,2,4,4-tetramethyl-1-piperidinyloxy (TEMPO) concentrations (0.02,0.08 M), the main source of radicals is the hydrogen abstraction of the Mayo dimer by TEMPO [with the kinetic constant of hydrogen abstraction (kh)]. At higher TEMPO concentrations ([N,] > 0.1 M), this reaction is still dominant, but radical generation by the direct attack against styrene by TEMPO, with kinetic constant of addition kad, also becomes relevant. From previous experimental data and simulations, initial estimates of kh , 1 and kad , 6 × 10,7 L mol,1 s,1 are obtained at 125 °C. From the induction period to the polymerization regime, there is an abrupt change in the dominant mechanism generating radicals because of the sudden decrease in the nitroxide radicals. Under induction-period conditions, the simulations confirm the validity of the quasi-steady-state assumption (QSSA) for the Mayo dimer in this regime; however, after the induction period, the QSSA for the dimer is not valid, and this brings into question the scientific basis of the well-known expression kth[M]3 (where [M] is the monomer concentration and kth is the kinetic constant of autoinitiation) for the autoinitiation rate in styrene polymerization. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 6962-6979, 2006 [source] Luminol chemiluminescence catalysed by colloidal platinum nanoparticlesLUMINESCENCE: THE JOURNAL OF BIOLOGICAL AND CHEMICAL LUMINESCENCE, Issue 2 2007Sheng-Liang Xu Abstract Platinum colloids prepared by the reduction of hexachloroplatinic acid with citrate in the presence of different stabilizers were found to enhance the chemiluminescence (CL) of the luminol,H2O2 system, and the most intensive CL signals were obtained with citrate-protected Pt colloids synthesized with citrate as both a reductant and a stabilizer. Light emission was intense and reproducible. Transmission electron microscopy and X-ray photoelectron spectroscopy studies were conducted before and after the CL reaction to investigate the possible CL enhancement mechanism. It is suggested that this CL enhancement is attributed to the catalysis of platinum nanoparticles, which could accelerate the electron-transfer process and facilitate the CL radical generation in aqueous solution. The effects of Pt colloids prepared by the hydroborate reduction were also investigated. The application of the luminol,H2O2,Pt colloids system was exploited for the determination of compounds such as uric acid, ascorbic acid, phenols and amino acids. Copyright © 2006 John Wiley & Sons, Ltd. [source] Prediction of Chain Length Distribution of Polystyrene Made in Batch Reactors with Bifunctional Free-Radical Initiators Using Dynamic Monte Carlo SimulationMACROMOLECULAR REACTION ENGINEERING, Issue 3 2007Ibrahim M. Maafa Abstract The objective of this paper is to present a dynamic Monte Carlo model that is able to simulate the polymerization of styrene with bifunctional free-radical initiators in a batch reactor. The model can predict the dynamic evolution of the chain length distribution of polystyrene in the reactor. The model includes all relevant polymerization mechanistic steps, including chemical and thermal radical generation, and diffusion-controlled termination. The model was applied to styrene polymerization and the Monte Carlo estimates for chain length averages were compared to those obtained with the method of moments. Excellent agreement was obtained between the two methods. Although styrene polymerization was used as a case study, the proposed methodology can be easily extended to any other polymer type made by free-radical polymerization. [source] Hyperglycemia Stimulates a Sustained Increase in Hydraulic Conductivity In Vivo without Any Change in Reflection CoefficientMICROCIRCULATION, Issue 7 2007RACHEL M. PERRIN ABSTRACT Objective: Increased microvascular permeability contributes to the development of diabetic microvascular complications and diabetic vasculopathy is correlated with blood glucose levels. The mechanisms underlying increased permeability, however, are poorly understood. Methods: The Landis-Michel technique was used to measure water permeability (hydraulic conductivity, Lp) and macromolecular permeability (reflection coefficient, ,) of exchange capillaries in frogs and rats. Results: Dialysed normoglycemic plasma from diabetic patients had no effect on Lp. The same plasma with 20 mM glucose increased hydraulic conductivity from (mean ± SEM × 10,7 cm · s,1· cm H2O,1) 5.73 ± 2.01 to 13.09 ± 2.67 (P < .01). Nondiabetic control plasma did not affect Lp, but addition of 20 mM glucose increased Lp to a similar degree. The effect of glucose alone was examined. Glucose at 20 mM increased Lp, from 2.82 ± 0.61 to 4.71 ± 1.35 × 10, 7 cm · s, 1· cm H2O,1 (P = .002, n = 13). A similar increase was seen in rat mesenteric microvessels, from 1.04 ± 0.40 in control perfusions to 2.18 ± 0.56, P < .05. The microvascular macromolecular reflection coefficient in all the above experiments was unaltered. The use of specific inhibitors indicated that the glucose-induced increased Lp did not appear to be mediated through protein kinase C (PKC), free radical generation, glucose metabolism, or albumin glycation. Conclusions: These data suggest that hyperglycemia induced increased apparent protein permeability may be secondary to a glucose-mediated change in macromolecular convective flux rather than any change in protein permeability per se. The authors speculate that the increased microvascular permeability to water in vivo is mediated by direct interaction of glucose with the endothelial cells (perhaps with the glycocalyx). [source] Plasma-Assisted Atomic Layer Deposition of Al2O3 at Room TemperaturePLASMA PROCESSES AND POLYMERS, Issue S1 2009Tommi O. Kääriäinen Abstract A new design of plasma source has been used for the plasma-assisted atomic layer deposition (PA-ALD) of Al2O3 films at room temperature. In this PA-ALD reactor the plasma is generated by capacitive coupling directly in the deposition chamber adjacent to the substrate but can be separated from it by a grid to reduce the ion bombardment while maintaining the flow of radicals directly to the substrate surface. During the ALD cycle a mixture of nitrogen and argon was introduced into the reactor to act as a purge gas between precursor pulses and to facilitate the generation of a plasma during the plasma cycle. Sequential exposures of TMA and excited O2 precursors were used to deposit Al2O3 films on Si(100) substrates. A plasma discharge was activated during the oxygen gas pulse to form radicals in the reactor space. The experiments showed that the growth rate of the film increased with increasing plasma power and with increasing O2 pulse length before saturating at higher power and longer O2 pulse length. The growth rate saturated at the level of 1.78 Å·cycle,1. EDS analysis showed that the films were oxygen rich and had carbon as an impurity. This can be explained by the presence of bonds between hydrocarbons from the unreacted TMA precursor and excess oxygen in the film. ATR-FTIR spectroscopy measurements indicated a change in growth mechanism when the distance between the location of the radical generation and the substrate was varied. A similar effect was observed with the use of different plasma power levels. [source] Methylmalonic acidaemia leads to increased production of reactive oxygen species and induction of apoptosis through the mitochondrial/caspase pathway,THE JOURNAL OF PATHOLOGY, Issue 4 2007E Richard Abstract Methylmalonic acidaemia (MMA) is a heterogeneous group of rare genetic metabolic disorders caused by defects related to intracellular cobalamin (vitamin B12) metabolism. Increasing evidence has emerged suggesting that free radical generation is involved in the pathophysiology of neurodegenerative diseases, including some inborn errors of metabolism. We have previously identified in MMA patients several differentially expressed proteins involved in oxidative stress [mitochondrial superoxide dismutase (MnSOD) and mitochondrial glycerophosphate dehydrogenase (mGPDH)] and apoptosis by a proteomic approach. We have now extensively evaluated various parameters related to oxidative stress and apoptosis in cultured fibroblasts from a spectrum of patients with methylmalonic acidaemia. Fibroblasts from several MMA patients showed a significant increase in intracellular reactive oxygen species (ROS) content and in MnSOD expression level with respect to controls, suggesting a cellular response to intrinsic ROS stress. Moreover, we have demonstrated, using siRNA, that mGPDH is an important ROS generator in MMA patients. Cells from patients with MMA had a higher rate of apoptosis than those of controls and there was evidence that this process primarily involves the mitochondrial/caspase-dependent pathway. ROS level,phenotype correlation revealed that patients with severe neonatal cblB disorder had elevated intracellular ROS content. These findings support the possible role of oxidative stress in the pathophysiology of methylmalonic acidaemia. Copyright © 2007 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. [source] Repeated bouts of aerobic exercise lead to reductions in skeletal muscle free radical generation and nuclear factor ,B activationTHE JOURNAL OF PHYSIOLOGY, Issue 16 2008Susan V. Brooks Chronic exercise improves endurance and skeletal muscle oxidative capacity. Despite the potential importance of reactive oxygen species (ROS) generated during exercise as regulators of these adaptations, the effect of repeated bouts of aerobic exercise on ROS generation by skeletal muscles during contractions has not been examined. Our aim was to establish the impact of repeated treadmill running exercise on muscle ROS generation and activation of redox-sensitive transcription factors. Following 8 weeks of treadmill running, mice displayed an improvement in running speed that was associated with an enhanced ability of gastrocnemius (GTN) muscles to maintain force during a protocol of isometric contractions. In contrast to GTN muscles of cage-sedentary (Sed) mice, muscles from exercised (Exer) mice did not release superoxide or nitric oxide during the isometric contractions. For male mice, basal levels of nuclear factor ,B (NF,B) and activator protein-1 (AP-1) DNA binding were increased by treadmill running, and the contraction-induced activation of NF,B and AP-1 observed in muscles of Sed mice was absent in Exer muscles. Also in contrast to Sed muscles, Exer muscles displayed no reductions in glutathione or protein thiol levels in response to contraction. Our observations of decreases for Exer compared with Sed muscles in contraction-induced (i) ROS generation, (ii) activation of redox-sensitive signalling pathways, and (iii) ROS stress suggest that exercise conditioning enhances the ability of skeletal muscle to readily and rapidly detoxify ROS and/or reduces ROS generation, providing protection from ROS-induced damage and reducing signals that might act to mediate further unnecessary adaptations. [source] Myo -inositol prevents oxidative damage, inhibits oxygen radical generation and increases antioxidant enzyme activities of juvenile Jian carp (Cyprinus carpio var. Jian)AQUACULTURE RESEARCH, Issue 15 2009Wei-Dan Jiang Abstract This study was conducted to evaluate the effects of dietary myo -inositol (MI) on the antioxidant status of juvenile Jian carp (Cyprinus carpio var. Jian). A total of 1050 Jian carp (22.28±0.07 g) were randomly distributed into seven groups of three replicates each, feeding diets containing graded levels of MI (163.5, 232.7, 384.2, 535.8, 687.3, 838.8 and 990.3 mg kg,1 diet) for 60 days. Results indicated that the malondialdehyde content was the lowest for fish fed diets containing ,384.2 mg MI kg,1, and the highest for fish fed the MI-unsupplemented basal diet (P<0.05). The protein carbonyl content was decreased with increasing dietary MI levels up to 535.8 mg kg,1 diet, and no differences were found with a further increase in the MI concentration. The anti-superoxide anion capacity (ASA) and anti-hydroxyl radical capacity (AHR) were increased with increasing MI levels up to 535.8 mg kg,1 diet, and plateaued thereafter. The superoxide dismutase and glutathione- S -transferase activities showed the same tendency with the ASA capacity. Catalase, glutathione peroxidase and glutathione reducase activities were improved with increasing MI levels up to 838.8, 384.2 and 687.3 mg kg,1 diet, respectively, and remained nearly constant thereafter. These results suggested that MI could inhibit oxygen radical generation, increase enzymatic antioxidant capacity and prevent oxidative damage of carp. Dietary MI requirements for ASA and AHR activities of juvenile Jian carp were 567.94 and 517.22 mg MI kg,1 diet respectively. [source] Free radical generation during the activation of hemolymph prepared from the homopteran Dactylopius coccusARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY (ELECTRONIC), Issue 1 2007F. Garcķa-Gil De Muńoz Abstract Superoxide anion (O,2) and nitric oxide (NO) generation in Dactylopius coccus hemolymph obtained by perfusion and activated with zymosan was studied. Activated hemolymph reduced 3-[4,5 dimethylthiazolil-2]-2,5-diphenyl tetrazolium bromide. This reduction was prevented by superoxide dismutase (SOD) indicating O,2 generation. This activity was dependent on temperature, and hemolymph incubated at 75°C lost its activity. Chromatocytes incubated with zymosan released their content and produced O,2. Activated hemolymph also produced NO and this activity was prevented in the presence of NG-nitro-L-arginine methyl ester, suggesting that nitric oxide synthase (NOS) might be present in D. coccus hemolymph. The probable source of O,2 in the D. coccus hemolymph is the anthraquinone oxidation, since commercial carminic dye produced O,2 during its oxidation by Agaricus bisporus tyrosinase. Gram+ Micrococcus luteus exposed to activated hemolymph were killed in vitro, and addition of NG-nitro-L-arginine methyl ester and D-Mannitol (a hydroxyl radical scavenger) prevented their killing. The cytotoxic effect produced by the activated hemolymph was not observed with the Gram, bacteria Serratia marcescens. These results suggest that D. coccus activated hemolymph generates reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI) that may limit M. luteus growth. Arch. Insect Biochem. Physiol. 65:20,28, 2007. © 2007 Wiley-Liss, Inc. [source] Effect of Nitric Oxide Synthase Inhibitors on Lipid Peroxide Formation in Liver Caused by Endotoxin ChallengeBASIC AND CLINICAL PHARMACOLOGY & TOXICOLOGY, Issue 4 2000Shuhei Sakaguchi This study investigated the effect of nitric oxide on lipid peroxide formation during endotoxaemia. Nitric oxide synthase inhibitors NG -monomethyl-L-arginine acetate (L-NMMA, 20 mg/kg, intravenously), NG -nitro-L-arginine-methyl ester (L-NAME, 10 mg/kg, intravenously), and NG -nitro-L-arginine (L-NA, 10 mg/kg, intravenously), and a relatively selective inducible nitric oxide synthase inhibitor aminoguanidine (10 mg/kg, intravenously), did not protect against endotoxin-induced death of mice. Superoxide dismutase activity in liver 18 hr after administration of endotoxin (6 mg/kg, intraperitoneally) to L-arginine analogues (L-NMMA, L-NAME, L-NA)-treated mice was lower than in mice treated with endotoxin alone, whereas the administration of L-arginine analogues increased xanthine oxidase activity in the livers of endotoxin-injected mice compared with mice treated with endotoxin alone. In mice treated with L-arginine analogues and aminoguanidine, the levels of non-protein sulfhydryl and lipid peroxide in liver 18 hr after endotoxin injection did not show significant differences from mice treated with endotoxin alone. L-Arginine analogues and aminoguanidine had little effect on lipid peroxide formation in liver caused by endotoxin. Treatment with aminoguanidine (300 ,M) significantly inhibited endotoxin-induced intracellular peroxide in J774A.1 cells, however, aminoguanidine did not affect endotoxin-induced cytotoxicity in J774A.1 cells. Our results clearly demonstrate that treatment with catalase (10 ,g/ml), D-mannitol (10 mM), or superoxide dismutase (100 U/ml), has little or no effect on nitric oxide production by endotoxin (1 ,g/ml)-activated J774A.1 cells. These findings suggest that nitric oxide is not crucial for lipid peroxide formation during endotoxaemia. Therefore, it is unlikely that nitric oxide plays a significant role in liver injury caused by free radical generation in endotoxaemia. [source] Differential effects of redox-cycling and arylating quinones on trans-plasma membrane electron transportBIOFACTORS, Issue 3 2008An S. Tan Abstract Cytotoxicity of quinones has been attributed to free radical generation and to arylation of cellular nucleophiles. For redox-cycling quinones, cell injury is associated with mitochondrial permeability transition, whereas arylating quinones directly depolarise the mitochondrial membrane and deplete ATP. Like mitochondrial electron transport, plasma membrane electron transport (PMET), plays a multifaceted role in cellular redox homeostasis but the effects of quinones on PMET are unknown. Here we investigate the effects of redox-cycling 2,3-dimethoxy-1,4-naphthoquinone (DMNQ), arylating 1,4-benzoquinone (BQ) and mixed mechanism 2-methyl-1,4-naphthoquinone (MNQ) on PMET, viability and growth of P815 mouse mastocytoma cells. BQ and MNQ rapidly and extensively inhibited PMET as determined by WST-1 /mPMS reduction (IC50 3.5-5 ,Mat 30 min) whereas the effects of DMNQ were less pronounced. In contrast, MTT reduction (cytosolic NADH dehydrogenase activity over 30 min) was weakly inhibited by BQ (IC50 20 ,M) but not by MNQ or DMNQ and cell viability was unaffected. Inhibition of WST-1/mPMS reduction by BQ and MNQ but not DMNQ was fully reversed by NAC. Treatment with DMNQ, MNQ and to a lesser extent BQ inhibited cell proliferation as determined by MTT reduction at 48 h. The effects of BQ and MNQ were reversed by NAC through covalent bonding to BQ and MNQ, but not DMNQ. These results show that arylating quinones are more potent inhibitors of PMET than pure redox-cycling quinones, but that redox-cycling quinones are more cytotoxic. [source] Oxidative Stress and Cardiovascular Disease: Antioxidants and Unresolved IssuesCARDIOVASCULAR THERAPEUTICS, Issue 2 2005Kamakshi Sachidanandam ABSTRACT Experimental and clinical studies suggest that oxidative stress contributes to the development and progression of cardiovascular disease. However, clinical trials with classic vitamin antioxidants failed to demonstrate any benefit in cardiovascular outcomes. Recent advances in our understanding of mechanisms involved in free radical generation reinstate that a more comprehensive approach targeting the prevention of reactive oxygen species (ROS) formation early in the disease process may prove beneficial. Experimental studies and reviews in oxidative stress were selected to provide a better understanding of the roles of the reactive species in the initiation and progression of cardiovascular disease (CVD). Clinical studies that evaluated the efficacy of several classes of antioxidants in CVD were included in the second part of this review to discuss future therapeutic guidelines based on currently available evidence. In conclusion, before a potential role for antioxidants in the treatment of CVD is eliminated, more carefully designed studies with classic as well as new antioxidants in well-defined patient populations are warranted to provide a definitive answer. [source] Lignin Chemistry: Biosynthetic Study and Structural Characterisation of Coniferyl Alcohol Oligomers Formed In Vitro in a Micellar EnvironmentCHEMISTRY - A EUROPEAN JOURNAL, Issue 20 2010Samantha Reale Dr. Abstract Model coniferyl alcohol lignin (the so-called dehydrogenative polymerisate, DHP) was produced in water under homogeneous conditions guaranteed by the presence of a micellised cationic surfactant. A complete study of the activity of the enzymatic system peroxidase/H2O2 under our reaction conditions was reported and all the reaction products up to the pentamer were characterised by 1H,NMR spectroscopy and ESI mass spectrometry. Our system, and the molecules that have been generated in it, represent a closer mimicry of the natural microenvironment since an enzyme, under micellar conditions, reproduces the cell system better than in buffer alone. On the basis of the oligomers structures a new biosynthetic perspective was proposed that focused attention on a coniferyl alcohol dimeric quinone methide as the key intermediate of the reaction. A formal, strictly alternate sequence of a radical and an ionic step underlines the reaction, thus generating ordered oligolignols structures. Alternatively to other model lignins, our olignols present a lower degree of radical coupling between oligomeric units. This offers a closer biosynthetic situation to the observation of a low rate of radical generation in the cell wall. [source] Early phase of reperfusion of human kidney allograft does not affect an erythrocyte anti-oxidative systemNEPHROLOGY, Issue 5 2006LESZEK DOMA SUMMARY: Background: Generation of reactive oxygen specimens is the basic mechanism leading to ischaemia/reperfusion injury of the kidney graft. Oxygen burst is a trigger for sophisticated biochemical changes leading to generation of oxygenated lipids and changes in microcirculation, which recruit recipient's neutrophils and contribute to delayed graft function. It has been shown that the free radicals generation correlates with the activity of anti-oxidative system. Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione (GSH) are involved in protection against free radicals. Aim: To examine the activity of erythrocyte anti-oxidative system during reperfusion of the transplanted kidney allograft. Methods: The study included 40 renal transplant recipients. Blood was taken from the iliac vein before transplantation and from the graft's renal vein immediately, as well as 2 and 4 min after total reperfusion. The authors assessed the process of reperfusion using ThermaCAM SC500 termovision camera. Spectrophotometric methods were used to measure superoxide dismutase, glutathione peroxidase and catalase activity as well as glutathione concentrations in erythrocytes. Results: There were no statistically significant differences in the activities of superoxide dismutase, catalase and glutathione peroxidase as well as glutathione concentrations during the first 4 min after total graft reperfusion. Nevertheless, there was a positive correlation between the activity of superoxide dismutase and glutathione peroxidase. Conclusion: The results suggest that the erythrocyte anti-oxidative system is stable during the early phase after reperfusion. An association between some anti-oxidative enzymes was noted. [source] | ||||||||||||||||||||||||||||