ascorbic-acid and 6-hydroxy-2-5-7-8-tetramethylchroman-2-carboxylic-acid

ascorbic-acid has been researched along with 6-hydroxy-2-5-7-8-tetramethylchroman-2-carboxylic-acid* in 194 studies

Reviews

2 review(s) available for ascorbic-acid and 6-hydroxy-2-5-7-8-tetramethylchroman-2-carboxylic-acid

ArticleYear
The antioxidative potential of melatonin in the skin.
    Current problems in dermatology, 2001, Volume: 29

    Topics: Animals; Antioxidants; Ascorbic Acid; Chromans; Dose-Response Relationship, Drug; Dose-Response Relationship, Radiation; Free Radical Scavengers; Free Radicals; Glycolates; Humans; Interleukin-3; Leukocytes; Luminescent Measurements; Melatonin; Mice; Mice, Hairless; Oxidative Stress; Oxygen; Radiation-Protective Agents; Reactive Oxygen Species; Respiratory Burst; Singlet Oxygen; Skin; Skin Aging; Ultraviolet Rays

2001
Neuroprotection by antioxidants in models of global and focal ischemia.
    Annals of the New York Academy of Sciences, 1994, Nov-17, Volume: 738

    Topics: Animals; Antioxidants; Ascorbic Acid; Brain Ischemia; Chromans; Corpus Striatum; Hippocampus; Ischemic Attack, Transient; Male; Neurons; Rats; Rats, Wistar; Reperfusion Injury; Structure-Activity Relationship

1994

Other Studies

192 other study(ies) available for ascorbic-acid and 6-hydroxy-2-5-7-8-tetramethylchroman-2-carboxylic-acid

ArticleYear
Enhancement of chemical stability of curcumin-enriched oil-in-water emulsions: Impact of antioxidant type and concentration.
    Food chemistry, 2020, Aug-01, Volume: 320

    Curcumin is claimed to have many health benefits, but it has low chemical stability. In this study, the influence of food-grade antioxidants on the chemical degradation of curcumin-enriched oil-in-water emulsions was examined. The curcumin degradation rate and extent depended on antioxidant type. The water-soluble antioxidants were more effective at protecting curcumin from degradation than the oil-soluble ones, which may have been because curcumin degrades faster in water than in oil. Interestingly, the amphiphilic antioxidant was almost as effective as the water-soluble ones. The oil-soluble antioxidant actually slightly promoted curcumin degradation. In summary, curcumin retention after storage declined in the following order: 82.6% (Trolox) ~82.2% (ascorbic acid) >79.5% (ascorbyl palmitate) ≫57.9% (control) >52.7% (α-tocopherol). The effectiveness of ascorbic acid in stabilizing curcumin increased as its concentration was raised (0-300 μM). Our results may facilitate the creation of curcumin-enriched foods and beverages with enhanced bioactivity.

    Topics: alpha-Tocopherol; Antioxidants; Ascorbic Acid; Chromans; Curcumin; Emulsions; Oils; Solubility; Water

2020
Heterocyclic β-keto sulfide derivatives of carvacrol: Synthesis and copper (II) ion reducing capacity.
    Bioorganic & medicinal chemistry letters, 2019, 10-01, Volume: 29, Issue:19

    Sixteen β-keto sulfide derivatives of carvacrol (4-19) incorporating phenyl or N, O and S heterocyclic moieties were synthesized in three steps. The relationships between heterocyclic structure and cupric, Cu(II), ion reducing antioxidant capacity (CUPRAC) were examined. Nine of the compounds (8-9 and 13-19) showed better CUPRAC activity than trolox at neutral pH, with trolox equivalent antioxidant capacity (TEAC) coefficients ranging between 1.20 and 1.75. Two derivatives (11-12) showed comparable reducing capacity to trolox, with TEAC values of 0.95 for 11 and 1.02 for 12. Compounds 8-9 and 11-19 were more effective at reducing the Cu(II) ion than ascorbic acid and the parent compound, carvacrol. The most effective antioxidants were those containing an oxadiazole, thiadiazole or triazole moiety. In particular, the methyl thiadiazole derivative (15) had the highest Cu(II) ion reducing capacity, with a TEAC coefficient of 1.73.

    Topics: Antioxidants; Chromans; Copper; Cymenes; Heterocyclic Compounds; Molecular Structure; Sulfides

2019
Suppression of riboflavin-sensitized singlet oxygen generation by l-ascorbic acid, 3-O-ethyl-l-ascorbic acid and Trolox.
    Journal of photochemistry and photobiology. B, Biology, 2019, Volume: 191

    Riboflavin (RF), a water-soluble vitamin B

    Topics: Antioxidants; Ascorbic Acid; Chromans; Humans; Kinetics; Oxygen; Photosensitizing Agents; Riboflavin; Singlet Oxygen; Spectrum Analysis

2019
Bulk storage of mango (Mangifera indica L.) and pineapple (Ananas comosus L.) pulp: effect of pulping and storage temperature on phytochemicals and antioxidant activity.
    Journal of the science of food and agriculture, 2019, Aug-30, Volume: 99, Issue:11

    The effects of pulp extraction, thermal treatment and bulk storage of mango (Mangifera indica L.) and pineapple (Ananas comosus L.) pulps for 20 weeks at ambient (28 ± 2 °C) and cold (4 °C) temperatures on the bioactive phytochemicals and antioxidant activity were investigated.. The contents of total polyphenols in mango (10.5%) and pineapple (5.4%) increased during pulping. The ratio of the degradation rate constants (k. Bulk storage of mango and pineapple pulp under cold storage conditions (4 °C) is recommended as a better pulp preservation method than storage at ambient (28 ± 2 °C) temperature. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

    Topics: Ananas; Antioxidants; Ascorbic Acid; beta Carotene; Catechin; Chromans; Food Storage; Fruit; Mangifera; Phytochemicals; Plant Extracts; Polyphenols; Tannins; Temperature

2019
Biologically and chemically important hydrazino-containing imidazolines as antioxidant agents.
    Free radical research, 2018, Volume: 52, Issue:6

    Topics: Amidines; Animals; Ascorbic Acid; Benzhydryl Compounds; Biphenyl Compounds; Butylated Hydroxyanisole; Butylated Hydroxytoluene; Chromans; Erythrocytes; Free Radical Scavengers; Hydrazines; Hydrogen Peroxide; Imidazolines; Nitric Oxide; Picrates; Rats; Structure-Activity Relationship

2018
Biologically important hydrazide-containing fused azaisocytosines as antioxidant agents.
    Redox report : communications in free radical research, 2017, Volume: 22, Issue:6

    Two important classes of hydrazide-containing fused azaisocytosines were evaluated as possible antioxidants and characterised by UV spectroscopy.. 2,2-Diphenyl-1-picrylhydazyl (DPPH), nitric oxide (NO), hydrogen peroxide (H. The strongest DPPH scavengers were found to be 9, showing the potency superior to that of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate (PG) and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) and comparable to that of ascorbic acid (AA), and 6, revealing the antioxidant potency superior to that of BHA, BHT, PG and Trolox. In turn, 3 and 9 were the most promising NO scavengers, exhibiting the potency superior to that of BHA, BHT (3 and 9) and AA (3). The most potent H. Hydrazides 3, 6 and 9 with an antioxidant potential better or comparable to that of the well-known antioxidants are proposed as new antioxidant candidates.

    Topics: alpha-Tocopherol; Antioxidants; Ascorbic Acid; Butylated Hydroxytoluene; Chromans; Free Radical Scavengers; Hydrogen Peroxide; Lipid Peroxidation; Propyl Gallate

2017
Solutions to decrease a systematic error related to AAPH addition in the fluorescence-based ORAC assay.
    Analytical biochemistry, 2017, Feb-15, Volume: 519

    Oxygen radical absorbance capacity (ORAC) assay in 96-well multi-detection plate readers is a rapid method to determine total antioxidant capacity (TAC) in biological samples. A disadvantage of this method is that the antioxidant inhibition reaction does not start in all of the 96 wells at the same time due to technical limitations when dispensing the free radical-generating azo initiator 2,2'-azobis (2-methyl-propanimidamide) dihydrochloride (AAPH). The time delay between wells yields a systematic error that causes statistically significant differences in TAC determination of antioxidant solutions depending on their plate position. We propose two alternative solutions to avoid this AAPH-dependent error in ORAC assays.

    Topics: Amidines; Antioxidants; Ascorbic Acid; Biological Assay; Chromans; Fluorescence; Gallic Acid; Oxidants; Oxygen Radical Absorbance Capacity; Reactive Oxygen Species

2017
Vitamin C and Trolox decrease oxidative stress and hemolysis in cold-stored human red blood cells.
    Redox report : communications in free radical research, 2017, Volume: 22, Issue:6

    To investigate the effects of sodium ascorbate (SA) (5-3125 μM) and a combination of SA and Trolox (25 and 125 μM) on oxidative changes generated in red blood cells (RBCs) followed by up to 20 days refrigerated storage.. RBCs were isolated from CPD-preserved human blood. Percentage of hemolysis and extracellular activity of lactate dehydrogenase (LDH) were measured to assess the RBC membrane integrity. Lipid peroxidation (LPO), glutathione (GSH) and total antioxidant capacity (TAC) were quantified by thiobarbituric acid-reactive substances, Ellman's reagent and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate) [Formula: see text]-based assay, respectively.. SA failed to reduce the storage-induced hemolysis and RBC membrane permeability. Addition of SA resulted in a concentration-independent LPO inhibition and increased TAC. A combination of SA/Trolox supplemented to the RBC medium significantly inhibited hemolysis, LDH leakage, LPO, GSH depletion and enhanced TAC.. The effects of vitamin C action are closely concentration-dependent and may be modulated by a variety of compounds (e.g. Hb degradation products) released from RBCs during the prolonged storage, changing its properties from anti- to pro-oxidative. The two different class antioxidants (SA/Trolox) could possibly cooperate to be good potential RBC storage additives ensuring both antiradical and membrane stabilizing protection.

    Topics: Ascorbic Acid; Cells, Cultured; Chromans; Erythrocytes; Glutathione; Hemolysis; Humans; L-Lactate Dehydrogenase; Lipid Peroxidation; Oxidative Stress; Preservation, Biological

2017
Redox modulation of curcumin stability: Redox active antioxidants increase chemical stability of curcumin.
    Molecular nutrition & food research, 2016, Volume: 60, Issue:3

    Substantial studies have shown that curcumin, a dietary compound from turmeric, has beneficial effects on many diseases. However, curcumin rapidly degrades at physiological pH, making it difficult to interpret whether the observed actions of curcumin are from curcumin itself or its degradation products. Therefore, it is important to better understand the mechanisms involved in curcumin degradation and the roles of degradation in its biological actions.. Here, we show that a series of redox active antioxidants with diverse chemical structures, including gallic acid, ascorbate (vitamin C), tert-butylhydroquinone (TBHQ), caffeic acid, rosmarinic acid, and Trolox (a water-soluble analog of vitamin E), dramatically increased curcumin stability in phosphate buffer at physiological pH. When treated in basal cell culture medium in MC38 colon cancer cells, curcumin rapidly degraded with a half-life of several minutes and showed a weak antiproliferative effect; co-addition of antioxidants enhanced stability and antiproliferative effect of curcumin. Finally, co-administration of antioxidant significantly increased plasma level of curcumin in animal models.. Together, these studies strongly suggest that a redox-dependent mechanism plays a critical role in mediating curcumin degradation. In addition, curcumin itself, instead of its degradation products, is largely responsible for the observed biological actions of curcumin.

    Topics: Animals; Antioxidants; Ascorbic Acid; Caffeic Acids; Cell Line, Tumor; Cell Proliferation; Chromans; Cinnamates; Curcumin; Depsides; Drug Stability; Gallic Acid; Hydrogen-Ion Concentration; Hydroquinones; Male; Mice; Oxidation-Reduction; Rosmarinic Acid

2016
CFTR silencing in pancreatic β-cells reveals a functional impact on glucose-stimulated insulin secretion and oxidative stress response.
    American journal of physiology. Endocrinology and metabolism, 2016, Feb-01, Volume: 310, Issue:3

    Cystic fibrosis (CF)-related diabetes (CFRD) has become a critical complication that seriously affects the clinical outcomes of CF patients. Although CFRD has emerged as the most common nonpulmonary complication of CF, little is known about its etiopathogenesis. Additionally, whether oxidative stress (OxS), a common feature of CF and diabetes, influences CFRD pathophysiology requires clarification. The main objective of this study was to shed light on the role of the cystic fibrosis transmembrane conductance regulator (CFTR) in combination with OxS in insulin secretion from pancreatic β-cells. CFTR silencing was accomplished in MIN6 cells by stable expression of small hairpin RNAs (shRNA), and glucose-induced insulin secretion was evaluated in the presence and absence of the valuable prooxidant system iron/ascorbate (Fe/Asc; 0.075/0.75 mM) along with or without the antioxidant Trolox (1 mM). Insulin output from CFTR-silenced MIN6 cells was significantly reduced (∼ 70%) at basal and at different glucose concentrations compared with control Mock cells. Furthermore, CFTR silencing rendered MIN6 cells more sensitive to OxS as evidenced by both increased lipid peroxides and weakened antioxidant defense, especially following incubation with Fe/Asc. The decreased insulin secretion in CFTR-silenced MIN6 cells was associated with high levels of NF-κB (the major participant in inflammatory responses), raised apoptosis, and diminished ATP production in response to the Fe/Asc challenge. However, these defects were alleviated by the addition of Trolox, thereby pointing out the role of OxS in aggravating the effects of CFTR deficiency. Our findings indicate that CFTR deficiency in combination with OxS may contribute to endocrine cell dysfunction and insulin secretion, which at least in part may explain the development of CFRD.

    Topics: Adenosine Diphosphate; Adenosine Triphosphate; Animals; Antioxidants; Ascorbic Acid; Blotting, Western; Catalase; Cell Line, Tumor; Cell Survival; Chromans; Cystic Fibrosis Transmembrane Conductance Regulator; Gene Knockdown Techniques; Gene Silencing; Glucose; Glutathione; Glutathione Peroxidase; HEK293 Cells; Humans; Insulin; Insulin Secretion; Insulin-Secreting Cells; Iron; Lipid Peroxidation; Mice; Oxidative Stress; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Superoxide Dismutase; Trace Elements

2016
Dynamic nuclear polarization-magnetic resonance imaging at low ESR irradiation frequency for ascorbyl free radicals.
    Scientific reports, 2016, Feb-19, Volume: 6

    Highly water-soluble ubiquinone-0 (CoQ0) reacts with ascorbate monoanion (Asc) to mediate the production of ascorbyl free radicals (AFR). Using aqueous reaction mixture of CoQ0 and Asc, we obtained positively enhanced dynamic nuclear polarization (DNP)-magnetic resonance (MR) images of the AFR at low frequency (ranging from 515 to 530 MHz) of electron spin resonance (ESR) irradiation. The shape of the determined DNP spectrum was similar to ESR absorption spectra with doublet spectral peaks. The relative locational relationship of spectral peaks in the DNP spectra between the AFR (520 and 525 MHz), (14)N-labeled carbamoyl-PROXYL ((14)N-CmP) (526.5 MHz), and Oxo63 (522 MHz) was different from that in the X-band ESR spectra, but were similar to that in the 300-MHz ESR spectra. The ratio of DNP enhancement to radical concentration for the AFR was higher than those for (14)N-CmP, Oxo63, and flavin semiquinone radicals. The spectroscopic DNP properties observed for the AFR were essentially the same as those for AFR mediated by pyrroloquinoline quinone. Moreover, we made a success of in vivo DNP-MR imaging of the CoQ0-mediated AFR which was administered by the subcutaneous and oral injections as an imaging probe.

    Topics: Ascorbic Acid; Benzoquinones; Chromans; Electron Spin Resonance Spectroscopy; Free Radicals; Magnetic Resonance Imaging

2016
Measurement of Antioxidant Capacity by Electron Spin Resonance Spectroscopy Based on Copper(II) Reduction.
    Analytical chemistry, 2016, Apr-05, Volume: 88, Issue:7

    A new method is proposed for measuring the antioxidant capacity by electron spin resonance spectroscopy based on the loss of electron spin resonance signal after Cu(2+) is reduced to Cu(+) with antioxidant. Cu(+) was removed by precipitation in the presence of SCN(-). The remaining Cu(2+) was coordinated with diethyldithiocarbamate, extracted into n-butanol and determined by electron spin resonance spectrometry. Eight standards widely used in antioxidant capacity determination, including Trolox, ascorbic acid, ferulic acid, rutin, caffeic acid, quercetin, chlorogenic acid, and gallic acid were investigated. The standard curves for determining the eight standards were plotted, and results showed that the linear regression correlation coefficients were all high enough (r > 0.99). Trolox equivalent antioxidant capacity values for the antioxidant standards were calculated, and a good correlation (r > 0.94) between the values obtained by the present method and cupric reducing antioxidant capacity method was observed. The present method was applied to the analysis of real fruit samples and the evaluation of the antioxidant capacity of these fruits.

    Topics: Antioxidants; Ascorbic Acid; Caffeic Acids; Chlorogenic Acid; Chromans; Copper; Coumaric Acids; Electron Spin Resonance Spectroscopy; Fruit and Vegetable Juices; Gallic Acid; Linear Models; Oxidation-Reduction; Quercetin; Rutin; Thiocyanates; Time Factors

2016
"Cut-off" effect of antioxidants and/or probes of variable lipophilicity in microheterogeneous media.
    Food chemistry, 2016, Sep-01, Volume: 206

    The activities of two hydrophilic (ascorbic acid and Trolox) and two hydrophobic (α-tocopherol and BHT) antioxidants were measured by reaction with a series of 4-alkanoyloxyTEMPO radical probes 1 in buffered (pH 7), aqueous, micellar solutions of reduced Triton-X 100. In all cases, a cut-off effect was observed, in line with previous observations of the same effect for the partitioning of probe series 1 in this medium. These results support an interpretation of the cut-off effect in food emulsions, based on the "amphiphobic" nature of either the antioxidants or probes: competition between two molecular moieties, for the micellar hydrophobic core, tends to expose a reacting fragment differently to a more hydrophilic microenvironment, as the probe or antioxidant hydrophobicity increases.

    Topics: alpha-Tocopherol; Antioxidants; Ascorbic Acid; Butylated Hydroxytoluene; Chromans; Hydrophobic and Hydrophilic Interactions; Micelles; Oxidation-Reduction

2016
Antioxidative Effects of a Glucose-Cysteine Maillard Reaction Product on the Oxidative Stability of a Structured Lipid in a Complex Food Emulsion.
    Journal of food science, 2016, Volume: 81, Issue:12

    A glucose-cysteine Maillard reaction product (MRP) was produced and its antioxidant effects on lipid oxidation were determined for a structured-lipid enriched with polyunsaturated fatty acids in a complex emulsion. Trolox equivalent antioxidant capacities (TEAC) were determined for MRP heating intervals of 2, 4, and 6 h and were compared to α-tocopherol (TOC), MRP with TOC (TOC-MRP), and TOC with ascorbyl palmitate (TOC-AP). Emulsions were produced with total antioxidant additions of 0.02% of the oil, and lipid oxidation was monitored by peroxide and p-anisidine values over 56 d. Positive correlations between browning and heating time as well as TEAC were observed. Total TEAC values for the MRP at 6 h, TOC, TOC with the MRP at 6 h, and TOC-AP were 2.51, 3.87, 2.68, and 2.76 mg trolox eq/g, respectively. Oxidation results indicated a possible antioxidant effect for the MRP at 6 h on secondary oxidation for days 14 to 28. These results suggest that the MRP at 6 h could be useful in inhibiting secondary oxidation in complex emulsions.

    Topics: alpha-Tocopherol; Aniline Compounds; Antioxidants; Ascorbic Acid; Chromans; Cysteine; Emulsions; Fatty Acids, Unsaturated; Food; Glucose; Lipid Metabolism; Maillard Reaction; Oxidation-Reduction; Particle Size; Peroxides

2016
Effect of antioxidants on the genotoxicity of phenethyl isothiocyanate.
    Mutagenesis, 2015, Volume: 30, Issue:3

    Isothiocyanates are plant-derived compounds that may be beneficial in the prevention of certain chronic diseases. Yet, by stimulating the production of reactive oxygen species (ROS), isothiocyanates can be genotoxic. Whether antioxidants influence isothiocyanate-induced genotoxicity is unclear, but this situation was clarified appreciably herein. In HCT116 cells, phenethyl isothiocyanate (PEITC) increased ROS production, which was inhibited by N-acetylcysteine (NAC) and deferoxamine (DFO) but not by ascorbic acid (ASC) and trolox (TRX) that were found to be more potent radical scavengers. Surprisingly, ASC and TRX each intensified the DNA damage that was caused by PEITC, but neither ASC nor TRX by themselves caused any DNA damage. In contrast, NAC and DFO each not only attenuated PEITC-induced DNA damage but also attenuated the antioxidant-intensified, PEITC-induced DNA damage. To determine if the DNA damage could be related to possible changes in the major antioxidant defence system, glutathione (GSH) was investigated. PEITC lowered GSH levels, which was prevented by NAC, whereas ASC, TRX and DFO neither inhibited nor enhanced the GSH-lowering effect of PEITC. The GSH synthesis inhibitor, buthionine sulphoxime, intensified PEITC-induced DNA damage, although by itself buthionine sulphoxime did not directly cause DNA damage. The principal findings suggest that ASC and TRX make PEITC more genotoxic, which might be exploited in killing cancer cells as one approach in killing cancer cells is to extensively damage their DNA so as to initiate apoptosis.

    Topics: Apoptosis; Ascorbic Acid; Biphenyl Compounds; Chromans; DNA Damage; Drug Evaluation, Preclinical; Free Radical Scavengers; Glutathione; HCT116 Cells; HT29 Cells; Humans; Isothiocyanates; Mutagens; Picrates; Reactive Oxygen Species

2015
Genotoxic effect of ethacrynic acid and impact of antioxidants.
    Toxicology and applied pharmacology, 2015, Jul-01, Volume: 286, Issue:1

    It is known that ethacrynic acid (EA) decreases the intracellular levels of glutathione. Whether the anticipated oxidative stress affects the structural integrity of DNA is unknown. Therefore, DNA damage was assessed in EA-treated HCT116 cells, and the impact of several antioxidants was also determined. EA caused both concentration-dependent and time-dependent DNA damage that eventually resulted in cell death. Unexpectedly, the DNA damage caused by EA was intensified by either ascorbic acid or trolox. In contrast, EA-induced DNA damage was reduced by N-acetylcysteine and by the iron chelator, deferoxamine. In elucidating the DNA damage, it was determined that EA increased the production of reactive oxygen species, which was inhibited by N-acetylcysteine and deferoxamine but not by ascorbic acid and trolox. Also, EA decreased glutathione levels, which were inhibited by N-acetylcysteine. But, ascorbic acid, trolox, and deferoxamine neither inhibited nor enhanced the capacity of EA to decrease glutathione. Interestingly, the glutathione synthesis inhibitor, buthionine sulfoxime, lowered glutathione to a similar degree as EA, but no noticeable DNA damage was found. Nevertheless, buthionine sulfoxime potentiated the glutathione-lowering effect of EA and intensified the DNA damage caused by EA. Additionally, in examining redox-sensitive stress gene expression, it was found that EA increased HO-1, GADD153, and p21mRNA expression, in association with increased nuclear localization of Nrf-2 and p53 proteins. In contrast to ascorbic acid, trolox, and deferoxamine, N-acetylcysteine suppressed the EA-induced upregulation of GADD153, although not of HO-1. Overall, it is concluded that EA has genotoxic properties that can be amplified by certain antioxidants.

    Topics: Antioxidants; Ascorbic Acid; Buthionine Sulfoximine; Chromans; Cyclin-Dependent Kinase Inhibitor p21; Deferoxamine; DNA Damage; Ethacrynic Acid; Glutathione; HCT116 Cells; Heme Oxygenase-1; Humans; Mutagens; NF-E2-Related Factor 2; Reactive Oxygen Species; RNA, Messenger; Transcription Factor CHOP; Tumor Suppressor Protein p53

2015
Effects of antioxidants and pro-oxidants on cytotoxicity of dihydroartemisinin to Molt-4 human leukemia cells.
    Anticancer research, 2015, Volume: 35, Issue:4

    The objective of the present study was to investigate how oxidative status influences the effectiveness of cytotoxicity of artemisinin towards cancer cells. It is hypothesized that antioxidants would reduce, whereas pro-oxidants would enhance, cytotoxicity.. Molt-4 human leukemia cells were incubated with vitamins C, E, D3, dexamethasone, or hydrogen peroxide alone or in combination with dihydroartemisinin (DHA). Concentrations of these compounds studied were similar to those achievable by oral administration. Viable cell counts were performed before (0 h) and at, 24 and 48 h after treatment.. Vitamin C, vitamin D3, dexamethasone, and H2O2 caused significant Molt-4 cell death. Vitamin E caused an increase in Molt-4 cell growth. Vitamin C and vitamin D3 significantly interacted with DHA at the 48-h time point and with H2O2 at both 24-h and 48-h time points.. Cellular oxidative status could alter the potency of artemisinin in killing cancer cells.

    Topics: Antioxidants; Apoptosis; Artemisinins; Ascorbic Acid; Cell Line, Tumor; Cholecalciferol; Chromans; Dexamethasone; Humans; Hydrogen Peroxide; Leukemia; Oxidation-Reduction; Reactive Oxygen Species

2015
Solubilisation of a 2,2-diphenyl-1-picrylhydrazyl radical in water by β-cyclodextrin to evaluate the radical-scavenging activity of antioxidants in aqueous media.
    Chemical communications (Cambridge, England), 2015, May-14, Volume: 51, Issue:39

    A 2,2-diphenyl-1-picrylhydrazyl radical (DPPH˙) was successfully solubilised in water by β-cyclodextrin (β-CD). DPPH˙/β-CD thus obtained was demonstrated to be a powerful tool to evaluate the antioxidative activity of water-soluble antioxidants, such as ascorbate and Trolox, in aqueous buffer solutions.

    Topics: Ascorbic Acid; beta-Cyclodextrins; Biphenyl Compounds; Chromans; Free Radical Scavengers; Free Radicals; Picrates; Solubility; Water

2015
A new approach to elucidating repair reactions of resveratrol.
    Physical chemistry chemical physics : PCCP, 2015, Jun-07, Volume: 17, Issue:21

    The repair by co-antioxidants of the phenoxy radical of resveratrol, the famous health-preserving ingredient of red wine, is a key step of radical scavenging cascades in nature. To generate that radical, we employed 355 nm photoionization as a direct and selective access that reduces the chemical complexity and is equally applicable in organized phases; to monitor it, we used its hitherto unreported absorption in the red where no other species in our systems interfere. With this novel approach, we measured rate constants and H/D kinetic isotope effects for the repairs by ascorbate, trolox (a vitamin E analogue) and 4-aminophenol, and identified the mechanisms as one-step hydrogen abstractions. Cysteine and glutathione are unreactive. In micellar solution (SDS), the repair by ascorbate is much slower and involves only the hydrophilic phenoxy moieties protruding from the micelles. The new experimental strategy also led to a reevaluation of extinction coefficients, rate constants and mechanisms.

    Topics: Aminophenols; Antioxidants; Ascorbic Acid; Chromans; Kinetics; Phenols; Resveratrol; Stilbenes; Wine

2015
The Impact of Sonication on the Surface Quality of Single-Walled Carbon Nanotubes.
    Journal of pharmaceutical sciences, 2015, Volume: 104, Issue:8

    Sonication process is regularly adopted for dispersing single-walled carbon nanotubes (SWCNTs) in an aqueous medium. This can be achieved by either covalent functionalization of SWCNTs with strong acid or by noncovalent functionalization using dispersants that adsorb onto the surface of SWCNTs during dispersion. Because the dispersion process is usually performed using sonication, unintentional free radical formation during sonication process may induce covalent modification of SWCNT surface. Herein, we have systematically investigated the status of SWCNT surface modification under various sonication conditions using Raman spectroscopy. Comparing ID /IG (Raman intensities between D and G bands) ratio of SWCNTs under various sonication conditions suggests that typical sonication conditions (1-6 h bath sonication with sonication power between 3 and 80 W) in aqueous media do not induce covalent modification of SWCNT surface. In addition, we confirm that SWCNT dispersion with single-stranded DNA (ssDNA) involves noncovalent adsorption of ssDNA onto the surface of SWCNTs, but not covalent linkage between ssDNA and SWCNT surface.

    Topics: Adsorption; Algorithms; Antioxidants; Ascorbic Acid; Chromans; DNA, Single-Stranded; Drug Compounding; Drug Delivery Systems; Kinetics; Nanotubes, Carbon; Oligodeoxyribonucleotides; Oxidants; Sonication; Spectrum Analysis, Raman; Surface Properties

2015
Dietary Total Antioxidant Capacity and Colorectal Cancer in the Italian EPIC Cohort.
    PloS one, 2015, Volume: 10, Issue:11

    Colorectal cancer is the third most common cancer worldwide. Diet has been hypothesized as involved in colorectal cancer etiology, but few studies on the influence of total dietary antioxidant intake on colorectal cancer risk have been performed.. We investigated the association between colorectal cancer risk and the total antioxidant capacity (TAC) of the diet, and also of intake of selected antioxidants, in 45,194 persons enrolled in 5 centers (Florence, Naples, Ragusa, Turin and Varese) of the European Prospective Investigation into Cancer and Nutrition (EPIC) Italy study. TAC was estimated by the Trolox equivalent antioxidant capacity (TEAC) assay. Hazard ratios (HRs) for developing colorectal cancer, and colon and rectal cancers separately, adjusted for confounders, were estimated for tertiles of TAC by Cox modeling, stratifying by center.. Four hundred thirty-six colorectal cancers were diagnosed over a mean follow-up of 11.28 years. No significant association between dietary TAC and colorectal cancer incidence was found. However for the highest category of TAC compared to the lowest, risk of developing colon cancer was lower (HR: 0.63; 95% CI: 0.44-0.89, P trend: 0.008). By contrast, increasing TAC intake was associated with significantly increasing risks of rectal cancer (2nd tertile HR: 2.09; 95%CI: 1.19-3.66; 3rd tertile 2.48 95%CI: 1.32-4.66; P trend 0.007). Intakes of vitamin C, vitamin E, and ß-carotene were not significantly associated with colorectal cancer risk.. Further prospective studies are needed to confirm the contrasting effects of high total antioxidant intake on risk of colon and rectal cancers.

    Topics: Adult; Anthropometry; Antioxidants; Ascorbic Acid; beta Carotene; Chromans; Colorectal Neoplasms; Diet; Feeding Behavior; Female; Follow-Up Studies; Humans; Italy; Life Style; Male; Middle Aged; Nutrition Assessment; Proportional Hazards Models; Prospective Studies; Risk Factors; Vitamin E

2015
Lignans from the fruit of Schisandra glaucescens with antioxidant and neuroprotective properties.
    Journal of natural products, 2014, Jun-27, Volume: 77, Issue:6

    Two rare 7,8-seco-lignans (1, 2), three new lignan glycosides (3, 4a, 4b), and 10 known lignans (5-14) were isolated from the fruit of Schisandra glaucescens Diels. The absolute configurations of 1 and 2 were determined by comparing their experimental and calculated electronic circular dichroism spectra. The molecular structures of the new compounds (3, 4a, and 4b), including their absolute configurations, were determined using various spectroscopic methods and hydrolysis reactions. The antioxidant activities of the isolated compounds were tested using 2,2-diphenyl-1-picrylhydrazyl and ferric reducing antioxidant power assays. Compounds 4, 7, 8, 10, 11, and 12 exhibited antioxidant activities of varying potential in both assays. Of these compounds, 7 showed the strongest 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity, with IC50 values of 15.7 (150 μM DPPH) and 34.6 μM (300 μM DPPH), respectively, and 4, 12, and 7 displayed higher total antioxidant activities than Trolox in the ferric reducing antioxidant power assay. The neuroprotective effects of these compounds against Aβ25-35-induced cell death in SH-SY5Y cells were also investigated. Compounds 1, 2, 6, 7, 8, 11, and 12 exhibited statistically significant neuroprotective effects against Aβ25-35-induced SH-SY5Y cell death compared with the group treated only with Aβ25-35.

    Topics: Amyloid beta-Peptides; Antioxidants; Biphenyl Compounds; Chromans; Drugs, Chinese Herbal; Fruit; Glycosides; Inhibitory Concentration 50; Lignans; Molecular Structure; Neuroprotective Agents; Nuclear Magnetic Resonance, Biomolecular; Oxidation-Reduction; Peptide Fragments; Picrates; Plant Extracts; Schisandra

2014
Quality parameters, bioactive compounds and their correlation with antioxidant capacity of commercial fruit-based baby foods.
    Food science and technology international = Ciencia y tecnologia de los alimentos internacional, 2014, Volume: 20, Issue:7

    Comprehensive research is required to achieve the optimization of the antioxidant protection through baby foods, in particular, the commercially available fruit-based baby foods. This study investigated the physicochemical properties, ascorbic acid (AA), total carotenoids (TC), total phenolic content (TPC), trolox equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) of 23 different commercially available fruit-based baby foods. The main contribution to the total antioxidant capacity (trolox equivalent antioxidant capacity and oxygen radical absorbance capacity) was provided by ascorbic acid, followed by phenolic compounds, in accordance with a mathematical equation obtained from the data: TEAC = 245.906 + 7.727 × (AA) + 1.988 × (TPC) - 0.008 × (TC) and ORAC = 318.662 + 2.775 × (AA) - 0.531 × (TPC) - 0.073 × (TC). Moreover, a positive correlation (r = 0.346, p < 0.05) was found for oxygen radical absorbance capacity and trolox equivalent antioxidant capacity methods. Baby foods with different kind of fruits used as ingredients showed higher antioxidant capacity. Among the commercial baby foods analysed in this work, that treated by gentle steam cooking process had high levels of bioactive compounds and antioxidant capacity.

    Topics: Antioxidants; Ascorbic Acid; Carotenoids; Chromans; Commerce; Fruit; Humans; Infant; Infant Food; Phenols

2014
Nicotine mediates hypochlorous acid-induced nuclear protein damage in mammalian cells.
    Inflammation, 2014, Volume: 37, Issue:3

    Activated neutrophils secrete hypochlorous acid (HOCl) into the extracellular space of inflamed tissues. Because of short diffusion distance in biological fluids, HOCl-damaging effect is restricted to the extracellular compartment. The current study aimed at investigating the ability of nicotine, a component of tobacco and electronic cigarettes, to mediate HOCl-induced intracellular damage. We report, for the first time, that HOCl reacts with nicotine to produce nicotine chloramine (Nic-Cl). Nic-Cl caused dose-dependent damage to proliferating cell nuclear antigen (PCNA), a nuclear protein, in cultured mammalian lung and kidney cells. Vitamin C, vitamin E analogue (Trolox), glutathione, and N-acetyl-L-cysteine inhibited the Nic-Cl-induced PCNA damage, implicating oxidation in PCNA damage. These findings point out the ability of nicotine to mediate HOCl-induced intracellular damage and suggest antioxidants as protective measures. The results also raise the possibility that Nic-Cl can be created in the inflamed tissues of tobacco and electronic cigarette smokers and may contribute to smoking-related diseases.

    Topics: Acetylcysteine; Antioxidants; Ascorbic Acid; Cell Line; Cell Proliferation; Chromans; Glutathione; Humans; Hypochlorous Acid; Inflammation; Kidney; Lung; Neutrophils; Nicotine; Oxidation-Reduction; Proliferating Cell Nuclear Antigen

2014
Ascorbate reverses high glucose- and RAGE-induced leak of the endothelial permeability barrier.
    Biochemical and biophysical research communications, 2014, Feb-28, Volume: 445, Issue:1

    High glucose concentrations due to diabetes increase leakage of plasma constituents across the endothelial permeability barrier. We sought to determine whether vitamin C, or ascorbic acid (ascorbate), could reverse such high glucose-induced increases in endothelial barrier permeability. Human umbilical vein endothelial cells and two brain endothelial cell lines cultured at 25 mM glucose showed increases in endothelial barrier permeability to radiolabeled inulin compared to cells cultured at 5mM glucose. Acute loading of the cells for 30-60 min with ascorbate before the permeability assay prevented the high glucose-induced increase in permeability and decreased basal permeability at 5mM glucose. High glucose-induced barrier leakage was mediated largely by activation of the receptor for advanced glycation end products (RAGE), since it was prevented by RAGE blockade and mimicked by RAGE ligands. Intracellular ascorbate completely prevented RAGE ligand-induced increases in barrier permeability. The high glucose-induced increase in endothelial barrier permeability was also acutely decreased by several cell-penetrant antioxidants, suggesting that at least part of the ascorbate effect could be due to its ability to act as an antioxidant.

    Topics: Acetylcysteine; Animals; Antioxidants; Ascorbic Acid; Benzamides; Cell Line; Cell Membrane Permeability; Cells, Cultured; Chromans; Cyclic N-Oxides; Dithiothreitol; Dose-Response Relationship, Drug; Endothelial Cells; Glucose; Glycation End Products, Advanced; HMGB1 Protein; Human Umbilical Vein Endothelial Cells; Humans; Mice; Receptor for Advanced Glycation End Products; Receptors, Immunologic; Serum Albumin, Bovine; Spin Labels

2014
Geminate recombination as a photoprotection mechanism for fluorescent dyes.
    Angewandte Chemie (International ed. in English), 2014, May-26, Volume: 53, Issue:22

    Despite common presumption due to fast photodestruction pathways through higher excited states, we show that further improvement of photostability is still achievable with diffusion-limited photoprotection formulas. Single-molecule fluorescence spectroscopy reveals that thiolate ions effectively quench triplet states of dyes by photoinduced electron transfer. Interestingly, this reaction rarely yields a radical anion of the dye, but direct return to the ground state is promoted by an almost instantaneous back electron transfer (geminate recombination). This type of mechanism is not detected for commonly used reductants such as ascorbic acid and trolox. The mechanism avoids the formation of radical cations and improves the photostability of single fluorophores. We find that a combination of β-mercaptoethanol and classical reducing and oxidizing systems yields the best results for several dyes including Atto532 and Alexa568.

    Topics: Ascorbic Acid; Chromans; Electron Transport; Fluorescent Dyes; Mercaptoethanol; Oxidation-Reduction; Photobleaching; Sulfhydryl Compounds; Ultraviolet Rays

2014
Insights on antioxidant assays for biological samples based on the reduction of copper complexes-the importance of analytical conditions.
    International journal of molecular sciences, 2014, Jun-25, Volume: 15, Issue:7

    Total antioxidant capacity assays are recognized as instrumental to establish antioxidant status of biological samples, however the varying experimental conditions result in conclusions that may not be transposable to other settings. After selection of the complexing agent, reagent addition order, buffer type and concentration, copper reducing assays were adapted to a high-throughput scheme and validated using model biological antioxidant compounds of ascorbic acid, Trolox (a soluble analogue of vitamin E), uric acid and glutathione. A critical comparison was made based on real samples including NIST-909c human serum certified sample, and five study samples. The validated method provided linear range up to 100 µM Trolox, (limit of detection 2.3 µM; limit of quantification 7.7 µM) with recovery results above 85% and precision <5%. The validated developed method with an increased sensitivity is a sound choice for assessment of TAC in serum samples.

    Topics: Antioxidants; Ascorbic Acid; Blood Chemical Analysis; Chromans; Copper; Glutathione; High-Throughput Screening Assays; Humans; Oxidation-Reduction; Uric Acid

2014
Assessment of the phytochemical constituents and antioxidant activity of a bloom forming microalgae Euglena tuba.
    Biological research, 2014, Jun-04, Volume: 47

    Unstable generation of free radicals in the body are responsible for many degenerative diseases. A bloom forming algae Euglena tuba growing abundantly in the aquatic habitats of Cachar district in the state of Assam in North-East India was analysed for its phytochemical contents, antioxidant activity as well as free radical scavenging potentials.. Based on the ability of the extract in ABTS•+ radical cation inhibition and Fe3+ reducing power, the obtained results revealed the prominent antioxidant activity of the algae, with high correlation coefficient of its TEAC values to the respective phenolic and flavonoid contents. The extract had shown its scavenging activity for different free radicals and 41.89 ± 0.41 μg/ml, 5.83 ± 0.07 μg/ml, 278.46 ± 15.02 μg/ml and 223.25 ± 4.19 μg/ml were determined as the IC50 values for hydroxyl, superoxide, nitric oxide and hypochlorous acid respectively, which are lower than that of the corresponding reference standards. The phytochemical analysis also revealed that the phenolics, flavonoids, alkaloids, tannins and carbohydrates are present in adequate amount in the extract which was confirmed by HPLC analysis.. The results showed that 70% methanol extract of the algae possesses excellent antioxidant and free radical scavenging properties.

    Topics: Alkaloids; Animals; Antioxidants; Ascorbic Acid; Cell Extracts; Chromans; Chromatography, High Pressure Liquid; Euglena; Flavonoids; Free Radical Scavengers; Glucose; India; Lipid Peroxidation; Male; Methanol; Mice; Microalgae; Oxidation-Reduction; Phenols; Reducing Agents; Tannins

2014
Effect of N-acetylarginine, a metabolite accumulated in hyperargininemia, on parameters of oxidative stress in rats: protective role of vitamins and L-NAME.
    Cell biochemistry and function, 2014, Volume: 32, Issue:6

    In the present investigation, we initially evaluated the in vitro effect of N-acetylarginine on thiobarbituric acid-reactive substances (TBA-RS), total sulfhydryl content and on the activities of antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the blood, kidney and liver of rats. Results showed that N-acetylarginine, at a concentration of 5.0 μM, decreased the activity of CAT in erythrocytes, enhanced TBA-RS in the renal cortex, decreased CAT and SOD activities in the renal medulla and decreased CAT and increased SOD and GSH-Px activities in the liver of 60-day-old rats. Furthermore, we tested the influence of the antioxidants, trolox and ascorbic acid, as well as of the N(ω) -nitro-L-arginine methyl ester (L-NAME) on the effects elicited by N-acetylarginine on the parameters tested. Antioxidants and L-NAME prevented most of the alterations caused by N-acetylarginine on the oxidative stress parameters evaluated. Data indicate that oxidative stress induction is probably mediated by the generation of NO and/or ONOO(-) and other free radicals because L-NAME and antioxidants prevented the effects caused by N-acetylarginine in the blood, renal tissues and liver of rats. Our findings lend support to a potential therapeutic strategy for this condition, which may include the use of appropriate antioxidants for ameliorating the damage caused by N-acetylarginine.

    Topics: alpha-Tocopherol; Animals; Antioxidants; Arginine; Ascorbic Acid; Chromans; Hyperargininemia; Kidney Cortex; Kidney Medulla; Liver; NG-Nitroarginine Methyl Ester; Nitric Oxide Synthase; Oxidative Stress; Rats, Wistar; Sulfhydryl Compounds; Thiobarbituric Acid Reactive Substances; Vitamins

2014
Radical scavenging activity of antioxidants evaluated by means of electrogenerated HO radical.
    Talanta, 2014, Volume: 129

    A method is proposed and tested concerning the characterization of antioxidants by means of their reaction with electrogenerated HO radicals in galvanostatic assays with simultaneous O2 evolution, using a Pt anode fairly oxidized. The consumption of a set of species with antioxidant activity, ascorbic acid (AA), caffeic acid (CA), gallic acid (GA) and trolox (T), is described by a first order kinetics. The rate of the processes is limited by the kinetics of reaction with HO radicals and by the kinetics of charge transfer. Information regarding the scavenger activity of antioxidants is obtained by the relative value of the rate constant of the reaction between antioxidants and HO radicals, k(AO,HO)/k(O2). The number of HO radicals scavenged per molecule of antioxidant is also estimated and ranged from 260 (ascorbic acid) to 500 (gallic acid). The method is applied successfully in the characterization of the scavenger activity of ascorbic acid in a green-tea based beverage.

    Topics: Antioxidants; Ascorbic Acid; Beverages; Caffeic Acids; Chromans; Chromatography, High Pressure Liquid; Electrochemistry; Electrolysis; Food Analysis; Free Radical Scavengers; Gallic Acid; Hydroxyl Radical; Oxygen; Phenol; Tea

2014
Carotenoids are effective inhibitors of in vitro hemolysis of human erythrocytes, as determined by a practical and optimized cellular antioxidant assay.
    Journal of food science, 2014, Volume: 79, Issue:9

    β-Carotene, zeaxanthin, lutein, β-cryptoxanthin, and lycopene are liposoluble pigments widely distributed in vegetables and fruits and, after ingestion, these compounds are usually detected in human blood plasma. In this study, we evaluated their potential to inhibit hemolysis of human erythrocytes, as mediated by the toxicity of peroxyl radicals (ROO•). Thus, 2,2'-azobis (2-methylpropionamidine) dihydrochloride (AAPH) was used as ROO• generator and the hemolysis assay was carried out in experimental conditions optimized by response surface methodology, and successfully adapted to microplate assay. The optimized conditions were verified at 30 × 10(6) cells/mL, 17 mM of AAPH for 3 h, at which 48 ± 5% of hemolysis was achieved in freshly isolated erythrocytes. Among the tested carotenoids, lycopene (IC(50) = 0.24 ± 0.05 μM) was the most efficient to prevent the hemolysis, followed by β-carotene (0.32 ± 0.02 μM), lutein (0.38 ± 0.02 μM), and zeaxanthin (0.43 ± 0.02 μM). These carotenoids were at least 5 times more effective than quercetin, trolox, and ascorbic acid (positive controls). β-Cryptoxanthin did not present any erythroprotective effect, but rather induced a hemolytic effect at the highest tested concentration (3 μM). These results suggest that selected carotenoids may have potential to act as important erythroprotective agents by preventing ROO•-induced toxicity in human erythrocytes.

    Topics: Antioxidants; Ascorbic Acid; Carotenoids; Chromans; Cytoprotection; Erythrocytes; Hemolysis; Humans; Lutein; Peroxides; Quercetin

2014
The α-tocopherol derivative ESeroS-GS induces cell death and inhibits cell motility of breast cancer cells through the regulation of energy metabolism.
    European journal of pharmacology, 2014, Dec-15, Volume: 745

    Cancer cells are known to exhibit different hallmarks compared with normal cells. Therefore, targeting these features may improve the response to cancer therapy. In this study, we provided direct evidence that the α-tocopherol derivative ESeroS-GS inhibited the viability, migration, and invasion of breast cancer cells. ESeroS-GS induced cell death in different cancer cells in a dose-dependent manner but showed no significant effects on MCF-10A mammary epithelial cells. Although the ESeroS-GS-induced cell death in MDA-MB-231 breast cancer cells was accompanied with the generation of reactive oxygen species and the down regulation of mitochondrial membrane potential (MMP), no such effect on reactive oxygen species and MMP was seen in MCF-10A cells. Further studies indicated that ESeroS-GS down-regulated the expression of hexokinase II, SDH B, UQCRC2 and COX II in MDA-MB-231 cells but not in MCF-10A cells. The down-regulation of these enzymes accounts for the decreased oxidative phosphorylation (OXPHOS) and glycolysis in MDA-MB-231 cells upon ESeroS-GS treatment. We also found that sub-toxic concentration of ESeroS-GS treatment resulted in the impairment of F-actin cytoskeleton assembly and the consequently decreased migratory and invasive ability of MDA-MB-231 cells, which might be due to the inhibition of cellular energy metabolism. These results indicate that ESeroS-GS shows potential to become a novel anti-cancer agent by targeting the energy metabolism of cancer cells.

    Topics: Adenosine Triphosphate; alpha-Tocopherol; Animals; Antineoplastic Agents; Antioxidants; Ascorbic Acid; Benzopyrans; Breast Neoplasms; Cell Line; Cell Line, Tumor; Cell Movement; Cell Survival; Chromans; Dose-Response Relationship, Drug; Energy Metabolism; Female; Glycolysis; HeLa Cells; Humans; Indoles; Mammary Glands, Human; Membrane Potential, Mitochondrial; Mice; Neoplasm Invasiveness; NIH 3T3 Cells; Oxidative Phosphorylation; Reactive Oxygen Species; Vitamin E

2014
Polar paradox revisited: analogous pairs of hydrophilic and lipophilic antioxidants in linoleic acid emulsion containing Cu(II).
    Journal of the science of food and agriculture, 2013, Aug-15, Volume: 93, Issue:10

    Literature data are scarce on the activities of analogous pairs of hydrophilic and lipophilic antioxidants related to the 'polar paradox' distinguishing antioxidants based on their partitioning between lipids and water. The peroxidation of linoleic acid (LA) in the presence of either Cu(II) ions alone or Cu(II) ions combined with Trolox (TR), ascorbic acid (AA), hydroquinone (HQ) and gallic acid (GA), as hydrophilic antioxidants, or with α-tocopherol (TocH), ascorbyl palmitate (AP), tert-butyl hydroquinone (TBHQ) and propyl gallate (PG), as their respective lipophilic analogues, was investigated in aerated and incubated emulsions at 37 °C and pH 7.. LA peroxidation induced by Cu(II) followed pseudo-first-order kinetics with respect to the formation of primary (hydroperoxides) and secondary (aldehyde- and ketone-like) oxidation products, which were determined by ferric thiocyanate (Fe(III)-SCN) and thiobarbituric acid-reactive substances (TBARS) methods respectively. With the exception of TocH at certain concentrations, the tested compounds showed antioxidant behaviour depending on their polarities. The results were evaluated in the light of structure-activity relationships and the polar paradox.. The results of this study partly confirm the hypothesis that the polar paradox experiences limitations in oil-in-water emulsions and that its validity is also dependent on the concentrations of the antioxidants employed.

    Topics: alpha-Tocopherol; Antioxidants; Ascorbic Acid; Chromans; Copper; Emulsions; Gallic Acid; Hydrophobic and Hydrophilic Interactions; Hydroquinones; Iron; Kinetics; Linoleic Acid; Lipid Peroxidation; Peroxides; Propyl Gallate; Structure-Activity Relationship; Thiobarbituric Acid Reactive Substances; Thiocyanates

2013
Antioxidant enzymes mediate survival of breast cancer cells deprived of extracellular matrix.
    Cancer research, 2013, Jun-15, Volume: 73, Issue:12

    Metastasis by cancer cells relies upon the acquisition of the ability to evade anoikis, a cell death process elicited by detachment from extracellular matrix (ECM). The molecular mechanisms that ECM-detached cancer cells use to survive are not understood. Striking increases in reactive oxygen species (ROS) occur in ECM-detached mammary epithelial cells, threatening cell viability by inhibiting ATP production, suggesting that ROS must be neutralized if cells are to survive ECM-detachment. Here, we report the discovery of a prominent role for antioxidant enzymes, including catalase and superoxide dismutase, in facilitating the survival of breast cancer cells after ECM-detachment. Enhanced expression of antioxidant enzymes in nonmalignant mammary epithelial cells detached from ECM resulted in ATP elevation and survival in the luminal space of mammary acini. Conversely, silencing antioxidant enzyme expression in multiple breast cancer cell lines caused ATP reduction and compromised anchorage-independent growth. Notably, antioxidant enzyme-deficient cancer cells were compromised in their ability to form tumors in mice. In aggregate, our results reveal a vital role for antioxidant enzyme activity in maintaining metabolic activity and anchorage-independent growth in breast cancer cells. Furthermore, these findings imply that eliminating antioxidant enzyme activity may be an effective strategy to enhance susceptibility to cell death in cancer cells that may otherwise survive ECM-detachment.

    Topics: Adenosine Triphosphate; Animals; Antioxidants; Ascorbic Acid; Blotting, Western; Breast Neoplasms; Catalase; Catechin; Cell Adhesion; Cell Line; Cell Line, Tumor; Cell Survival; Chromans; Extracellular Matrix; Female; Humans; Mice; Mice, Nude; Reactive Oxygen Species; RNA Interference; Superoxide Dismutase; Tomography, X-Ray Computed; Xenograft Model Antitumor Assays

2013
Assessing and comparing the total antioxidant capacity of commercial beverages: application to beers, wines, waters and soft drinks using TRAP, TEAC and FRAP methods.
    Combinatorial chemistry & high throughput screening, 2013, Volume: 16, Issue:1

    This work measures and tries to compare the Antioxidant Capacity (AC) of 50 commercial beverages of different kinds: 6 wines, 12 beers, 18 soft drinks and 14 flavoured waters. Because there is no reference procedure established for this purpose, three different optical methods were used to analyse these samples: Total Radical trapping Antioxidant Parameter (TRAP), Trolox Equivalent Antioxidant Capacity (TEAC) and Ferric ion Reducing Antioxidant Parameter (FRAP). These methods differ on the chemical background and nature of redox system. The TRAP method involves the transfer of hydrogen atoms while TEAC and FRAP involves electron transfer reactions. The AC was also assessed against three antioxidants of reference, Ascorbic acid (AA), Gallic acid (GA) and 6-hydroxy-2,5,7,8-tetramethyl- 2-carboxylic acid (Trolox). The results obtained were analyzed statistically. Anova one-way tests were applied to all results and suggested that methods and standards exhibited significant statistical differences. The possible effect of sample features in the AC, such as gas, flavours, food colouring, sweeteners, acidity regulators, preservatives, stabilizers, vitamins, juice percentage, alcohol percentage, antioxidants and the colour was also investigated. The AC levels seemed to change with brand, kind of antioxidants added, and kind of flavour, depending on the sample. In general, higher ACs were obtained for FRAP as method, and beer for kind of sample, and the standard expressing the smaller AC values was GA.

    Topics: Antioxidants; Ascorbic Acid; Beer; Beverages; Carbonated Beverages; Chromans; Drinking Water; Food Analysis; Gallic Acid; Wine

2013
In vitro stimulation of oxidative stress by hypoxanthine in blood of rats: prevention by vitamins e plus C and allopurinol.
    Nucleosides, nucleotides & nucleic acids, 2013, Volume: 32, Issue:1

    We herein investigated the in vitro effect of hypoxanthine on the activities of antioxidant enzymes such as catalase (CAT), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) in erythrocytes, as well as on thiobarbituric acid-reactive substances (TBA-RS), in the plasma of rats. Results showed that hypoxanthine, when added to the incubation medium, enhanced CAT (10.0 μM), GSH-Px and SOD (8.5 μM and 10.0 μM) activities in erythrocytes of 15-day-old rats, reduced CAT activity (10.0 μM) and enhanced GSH-Px activity (10.0 μM) in erythrocytes of 30-day-old rats, reduced CAT activity (10.0 μM) and enhanced GSH-Px activity (8.5 μM and 10.0 μM) in erythrocytes of 60-day-old rats, as compared to controls. In addition, hypoxanthine (10.0 μM) enhanced TBA-RS levels in the plasma of 30- and 60-day old rats. Furthermore, we also tested the influence of allopurinol, trolox, and ascorbic acid on the effects elicited by hypoxanthine on the antioxidant enzymes and TBA-RS. Allopurinol and/or administration of antioxidants prevented most alterations caused by hypoxanthine in the oxidative stress parameters evaluated. Findings suggest that hypoxanthine alters antioxidant defenses and induces lipid peroxidation in the blood of rats; however, in the presence of allopurinol and antioxidants, some of these alterations in oxidative stress caused are prevented. Data indicate that, in humans, antioxidant administration might serve as a potential adjuvant therapy for ameliorating the damage caused by hypoxanthine.

    Topics: Allopurinol; Animals; Antioxidants; Ascorbic Acid; Catalase; Chromans; Erythrocytes; Glutathione Peroxidase; Hypoxanthines; Malondialdehyde; Oxidative Stress; Rats; Rats, Wistar; Superoxide Dismutase; Thiobarbituric Acid Reactive Substances; Vitamin E

2013
Racemization of the aspartic acid residue of amyloid-β peptide by a radical reaction.
    Bioscience, biotechnology, and biochemistry, 2013, Volume: 77, Issue:2

    Human amyloid-β peptide 1-42 (Aβ) was subjected to a radical reaction by using ascorbic acid and CuCl(2). The percentage of D-aspartic acid (D-Asp) after 24 h had increased to 6.69 ± 0.09%, this being comparable with the reported D-Asp concentration of purified core amyloids in Alzheimer's disease patients. This racemization was significantly inhibited by radical scavengers. L-Alanine was also racemized during the same reaction.

    Topics: Amyloid beta-Peptides; Ascorbic Acid; Aspartic Acid; Chromans; Copper; Free Radical Scavengers; Free Radicals; Humans; Mercaptoethanol; Peptide Fragments; Solutions; Stereoisomerism

2013
Trolox enhances curcumin's cytotoxicity through induction of oxidative stress.
    Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 2012, Volume: 29, Issue:3-4

    Curcumin, a natural polyphenol in the spice turmeric, has been found to exhibit anticancer activity. Although curcumin is generally considered an antioxidant, it is also able to elicit apoptosis through the generation of ROS, thereby functioning as a pro-oxidant in cancer cells. The present study investigated the effects of antioxidant pretreatment on curcumin-induced cytotoxicity in the human cancer cell lines A2780, MCF-7, and MDA-MB-231. Cytotoxicity was enhanced by trolox, vitamin C or vitamin E; trolox, a water soluble vitamin E derivative, was the most potent. The combination of curcumin (10 μM) and trolox (10-50 μM) induced apoptosis of cancer cells as evidenced by PARP cleavage and caspase-3 activation. Furthermore, expression of the pro-apoptotic protein Bad was up-regulated and expression of the anti-apoptotic proteins Bcl-2 and Bcl-xl was down-regulated in cells that had been treated with trolox plus curcumin. ROS generation was detected in curcumin-treated cells and was significantly enhanced when cells were treated with trolox plus curcumin. Exogenous catalase or SOD1 did not alter cytotoxicity, while over-expression of either catalase or SOD1 did, pointing to the importance of intracellular hydrogen peroxide generation in cell killing. In conclusion, we demonstrated for the first time that antioxidants such as trolox can potentiate cancer cell killing by curcumin, a finding which may help in the development of novel drug combination therapies.

    Topics: Antineoplastic Agents, Phytogenic; Antineoplastic Combined Chemotherapy Protocols; Antioxidants; Apoptosis; Ascorbic Acid; bcl-Associated Death Protein; Caspase 3; Cell Line, Tumor; Cell Survival; Chromans; Curcumin; Dose-Response Relationship, Drug; Enzyme Activation; Gene Expression Regulation, Neoplastic; Humans; Oxidative Stress; Poly(ADP-ribose) Polymerases; Proteolysis; Proto-Oncogene Proteins c-bcl-2; Reactive Oxygen Species; Superoxide Dismutase; Superoxide Dismutase-1; Vitamin E

2012
Motor deficits are triggered by reperfusion-reoxygenation injury as diagnosed by MRI and by a mechanism involving oxidants.
    The Journal of neuroscience : the official journal of the Society for Neuroscience, 2012, Apr-18, Volume: 32, Issue:16

    The early antecedents of cerebral palsy (CP) are unknown but are suspected to be due to hypoxia-ischemia (H-I). In our rabbit model of CP, the MRI biomarker, apparent diffusion coefficient (ADC) on diffusion-weighted imaging, predicted which fetuses will develop postnatal hypertonia. Surviving H-I fetuses experience reperfusion-reoxygenation but a subpopulation manifested a continued decline of ADC during early reperfusion-reoxygenation, which possibly represented greater brain injury (RepReOx). We hypothesized that oxidative stress in reperfusion-reoxygenation is a critical trigger for postnatal hypertonia. We investigated whether RepReOx predicted postnatal neurobehavior, indicated oxidative stress, and whether targeting antioxidants at RepReOx ameliorated motor deficits, which included testing of a new superoxide dismutase mimic (MnTnHex-2-PyP). Rabbit dams, 79% gestation (E25), were subjected to 40 min uterine ischemia. Fetal brain ADC was followed during H-I, immediate reperfusion-reoxygenation, and 4-72 h after H-I. Endpoints were postnatal neurological outcome at E32, ADC at end of H-I, ADC nadir during H-I and reperfusion-reoxygenation, and area under ADC curve during the first 20 min of reperfusion-reoxygenation. Antioxidants targeting RepReOx were administered before and/or after uterine ischemia. The new MRI-ADC biomarker for RepReOx improved prediction of postnatal hypertonia. Greater superoxide production, mitochondrial injury, and oligodendroglial loss occurred in fetal brains exhibiting RepReOx than in those without. The antioxidants, MnTnHex-2-PyP and Ascorbate and Trolox combination, significantly decreased postnatal motor deficits and extent of RepReOx. The etiological link between early injury and later motor deficits can thus be investigated by MRI, and allows us to distinguish between critical oxidative stress that causes motor deficits and noncritical oxidative stress that does not.

    Topics: Age Factors; Animals; Animals, Newborn; Antioxidants; Ascorbic Acid; Benzimidazoles; Blood Flow Velocity; Brain; Brain Mapping; Carbocyanines; Chromans; Diffusion Magnetic Resonance Imaging; Disease Models, Animal; Embryo, Mammalian; Female; Flow Cytometry; Hypoxia-Ischemia, Brain; Ionophores; Laser-Doppler Flowmetry; Membrane Potential, Mitochondrial; Metalloporphyrins; Microvessels; Mitochondria; Movement Disorders; Muscle Hypertonia; O Antigens; Pregnancy; Rabbits; Reperfusion Injury; Superoxides; Time Factors; Valinomycin

2012
Amaranth dye in the evaluation of bleaching of cerium (IV) by antioxidants: application in food and medicinal plants.
    Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy, 2012, Volume: 95

    A simple, low-cost, sensitive, and diversely applicable spectrophotometric method for the determination of total antioxidant capacity of several medicinal plants and food has been developed. The method is based on the bleaching of cerium (IV) by antioxidants and dye in slightly acid medium at room temperature. The unbleached dye, imparting pink color to the solution, is measured at λ(max) 530 nm which is directly proportional to the antioxidant concentration. The method is reproducible, and the trolox equivalent antioxidant capacities (TEAC coefficients) of the tested antioxidant compounds were correlated with those found by reference method such as ABTS. The recommended method was applied for the determination of total antioxidant capacity of medicinal and food samples. The performance of the recommended method was evaluated in terms of Student's t-test and variance ratio F-test, which indicated the significance of proposed method over the reference method.

    Topics: Absorption; Amaranth Dye; Antioxidants; Ascorbic Acid; Calibration; Cerium; Chromans; Fluorescence Recovery After Photobleaching; Food Analysis; Gallic Acid; Photobleaching; Plants, Medicinal; Quercetin; Spectrophotometry

2012
Evaluation of antioxidant activity of vanillin by using multiple antioxidant assays.
    Biochimica et biophysica acta, 2011, Volume: 1810, Issue:2

    Vanillin, a compound widely used in foods, beverages, cosmetics and drugs, has been reported to exhibit multifunctional effects such as antimutagenic, antiangiogenetic, anti-colitis, anti-sickling, and antianalgesic effects. However, results of studies on the antioxidant activity of vanillin are not consistent.. We systematically evaluated the antioxidant activity of vanillin using multiple assay systems. DPPH radical-, galvinoxyl radical-, and ABTS(+)-scavenging assays, ORAC assay and an oxidative hemolysis inhibition assay (OxHLIA) were used for determining the antioxidant activity.. Vanillin showed stronger activity than did ascorbic acid and Trolox in the ABTS(+)-scavenging assay but showed no activity in the DPPH radical- and galvinoxyl radical-scavenging assays. Vanillin showed much stronger antioxidant activity than did ascorbic acid and Trolox in the ORAC assay and OxHLIA. In the ABTS(+)-scavenging assay, ORAC assay and OxHLIA, vanillin reacted with radicals via a self-dimerization mechanism. The dimerization contributed to the high reaction stoichiometry against ABTS(+) and AAPH-derived radicals to result in the strong effect of vanillin. Oral administration of vanillin to mice increased the vanillin concentration and the antioxidant activity in plasma. These data suggested that antioxidant activity of vanillin might be more beneficial than has been thought for daily health care.. Based on the results of the present study, we propose the addition of antioxidant capacity to the multifunctionality of vanillin.

    Topics: Animals; Antioxidants; Ascorbic Acid; Benzaldehydes; Benzhydryl Compounds; Benzothiazoles; Biphenyl Compounds; Chromans; Chromatography, High Pressure Liquid; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Erythrocytes; Free Radical Scavengers; Hemolysis; Mice; Molecular Structure; Oxidation-Reduction; Picrates; Sheep; Sulfonic Acids

2011
Effect of micronutrients on methylglyoxal-mediated in vitro glycation of albumin.
    Biological trace element research, 2011, Volume: 143, Issue:2

    Nonenzymatic glycation of long-lived proteins has been implicated in several complications related to age and diabetes. Dicarbonyl compounds such as methylglyoxal (MGO) have been identified as the predominant source for the formation of advanced glycation end-products (AGEs) in various tissues. We investigated the effect of 13 micronutrients on MGO-mediated in vitro glycation of bovine serum albumin (BSA), as formation of AGEs and protein carbonyls. BSA (10 mg/ml) was incubated at 37°C with 100 mM MGO for 24 hours, in presence of ascorbic acid, Trolox (water-soluble α-tocopherol analog), β-carotene, retinol, riboflavin, thiamin, folic acid, niacin, pyridoxine, zinc, iron, manganese, and selenium. Fluorescence was measured at the wavelength pair of 370 and 440 nm as an index of the formation of AGEs and spectra were recorded for promising interactions at λex=280 nm and λex=370 nm. Within four standard antiglycating agents, aminoguanidine showed highest inhibitory response for BSA glycation followed by quercetin, gallic acid, and tannic acid. Promising antiglycation potential was seen for Trolox, riboflavin, Zn, and Mn as evidenced by decrease in the formation of AGEs and protein carbonyls.

    Topics: Animals; Ascorbic Acid; beta Carotene; Cattle; Chromans; Folic Acid; Glycation End Products, Advanced; Glycosylation; Iron; Manganese; Micronutrients; Pyridoxine; Pyruvaldehyde; Riboflavin; Selenium; Serum Albumin, Bovine; Thiamine; Vitamin A; Zinc

2011
Antioxidant and antibacterial activities of selected varieties of thai mango seed extract.
    Pakistan journal of pharmaceutical sciences, 2011, Volume: 24, Issue:1

    This study reports the antioxidant and antibacterial activities of four fresh mango seed extracts from Thai varieties. Total phenol contents determined by the Folin-ciocalteu method revealed the highest values to be in MKE, Chok-a-nan variety (399.8 mgGAE/g extract) and MSE of Nam-dok-mai variety (377.2 mgGAE/g extract). Both extracts showed potent ABTS˙+ radical and DPPH˙ radical scavenging activities with the lower half inhibition concentration (IC50) values than those of the reference compounds; vitamin C, trolox and BHA, respectively. Their antioxidant property of MSE and MKE is strongly correlated with the total phenol contents (r=0.98 and 0.98, respectively). When combined the MSE and MKE of the Fah-lun variety showed the strongest antioxidant activity. All mango seed extracts showed interesting antibacterial activity against both gram positive and gram negative bacteria as determined by disc diffusion method. The most sensitive pathogenic strain inhibited by all extracts (especially Kaew variety) was Pseudomonas aeruginosa ATCC 27853. This work suggests potential applications for practical uses of mango seed extracts from Thai varieties, as sources of antioxidant and antibacterial agents.

    Topics: Anti-Bacterial Agents; Antioxidants; Ascorbic Acid; Bacteria; Benzothiazoles; Biphenyl Compounds; Chromans; Diffusion; Free Radical Scavengers; Indicators and Reagents; Mangifera; Microbial Sensitivity Tests; Phenols; Picrates; Plant Extracts; Seeds; Sulfonic Acids; Thailand

2011
Evaluation of free-radical quenching properties of standard Ayurvedic formulation Vayasthapana Rasayana.
    BMC complementary and alternative medicine, 2011, May-12, Volume: 11

    Cellular damage induced by free-radicals like Reactive Oxygen and Nitrogen Species (ROS and RNS) has been implicated in several disorders and diseases, including ageing. Hence naturally occurring anti-oxidant rich-herbs play a vital role in combating these conditions. The present study was carried out to investigate the in vitro free-radical quenching capacity of a known Ayurvedic poly-herbal formulation called Vayasthapana Rasayana.. Methanol extracts of Vayasthapana Rasayana formulation (VRF) were studied for in vitro total antioxidant activity along with phenolic content and reducing power. In vitro assays like DPPH, FRAP, ABTS scavenging to evaluate radical quenching potential were performed.. The formulation has shown 94% at 0.1 mg/ml DPPH free-radical scavenging activity as against 84% at 0.1 mg/ml for standard ascorbic acid (IC₅₀ value 5.51 μg/ml for VRF and 39 μg/ml for standard). It has a significant higher ferric reducing potential also (OD 0.87 at 700 nm & 0.21 at 0.1 mg/ml for VRF and standard, respectively). The total phenolic content (gallic acid equivalent) of the VRF is 8.3 mg per g of dry mass. Total antioxidant capacity of the formulation, estimated by FRAP was 1150 ± 5 μM Fe(II)/g dry mass. ABTS radical scavenging activity of VRF was 69.55 ± 0.21% at 100 μg/ml concentration with a IC50 value of 69.87 μg/ml as against 9% and 95% by ascorbic acid and Trolox (at 70.452 μg/ml and 0.250 μg/ml concentrations, respectively).. In Indian traditional Ayurvedic system, use of VRF is in regular practice for mainly combating age-related disorders and diseases as many of the components of the Rasayana are known for their free-radical scavenging activity. This study has validated the potential use of VRF as an anti-oxidant to fight age-related problems.

    Topics: Aging; Antioxidants; Ascorbic Acid; Benzothiazoles; Biphenyl Compounds; Chromans; Ferric Compounds; Humans; Magnoliopsida; Medicine, Ayurvedic; Oxidative Stress; Phenols; Picrates; Plant Extracts; Reference Standards; Sulfonic Acids; Thiazoles

2011
Ramalin, a novel nontoxic antioxidant compound from the Antarctic lichen Ramalina terebrata.
    Phytomedicine : international journal of phytotherapy and phytopharmacology, 2011, Nov-15, Volume: 18, Issue:14

    Ramalin (γ-glutamyl-N'-(2-hydroxyphenyl)hydrazide), a novel compound, was isolated from the methanol-water extract of the Antarctic lichen Ramalina terebrata by several chromatographic methods. The molecular structure of ramalin was determined by spectroscopic analysis. The experimental data showed that ramalin was five times more potent than commercial butylated hydroxyanisole (BHA) in scavenging 1-diphenyl-2-picryl-hydazil (DPPH) free radicals, 27 times more potent in scavenging 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid free radicals (ABTS(+)) than the vitamin E analogue, trolox, and 2.5 times more potent than BHT in reducing Fe(3+) to Fe(2+) ions. Similarly, ramalin was 1.2 times more potent than ascorbic acid in scavenging superoxide radicals and 1.25 times more potent than commercial kojic acid in inhibiting tyrosinase enzyme activity, which ultimately leads to whitening of skin cells. Ramalin showed no or very little cytotoxicity in human keratinocyte and fibroblast cells at its antioxidant concentration. Furthermore, ramalin was assessed to determine its antioxidant activity in vivo. One microgram per milliliter ramalin significantly reduced the released nitric oxide (NO) and 0.125 μg/ml ramalin reduced the produced hydrogen peroxide (H(2)O(2)) in LPS (lipopolysaccharide)-stimulated murine macrophage Raw264.7 cells. Considering all the data together, ramalin can be a strong therapeutic candidate for controlling oxidative stress in cells.

    Topics: Animals; Antarctic Regions; Antioxidants; Ascorbic Acid; Benzothiazoles; Biphenyl Compounds; Butylated Hydroxyanisole; Cell Line; Cell Survival; Chromans; Fibroblasts; Free Radicals; Fungal Proteins; Glutamates; Humans; Hydrogen Peroxide; Keratinocytes; Lichens; Mice; Molecular Structure; Monophenol Monooxygenase; Nitric Oxide; Picrates; Pyrones; Sulfonic Acids

2011
Homocysteine induces hypophosphorylation of intermediate filaments and reorganization of actin cytoskeleton in C6 glioma cells.
    Cellular and molecular neurobiology, 2010, Volume: 30, Issue:4

    In this study, we investigated the actions of high homocysteine (Hcy) levels (100 and 500 microM) on the cytoskeleton of C6 glioma cells. Results showed that the predominant cytoskeletal response was massive formation of actin-containing filopodia at the cell surface that could be related with Cdc42 activation and increased vinculin immunocontent. In cells treated with 100 microM Hcy, folic acid, trolox, and ascorbic acid, totally prevented filopodia formation, while filopodia induced by 500 microM Hcy were prevented by ascorbic acid and attenuated by folic acid and trolox. Moreover, competitive NMDA ionotropic antagonist DL-AP5 totally prevented the formation of filopodia in both 100 and 500 microM Hcy treated cells, while the metabotropic non-selective group I/II antagonist MCPG prevented the effect of 100 microM Hcy but only slightly attenuated the effect induced by of 500 microM Hcy on actin cytoskeleton. The competitive non-NMDA ionotropic antagonist CNQX was not able to prevent the effects of Hcy on the reorganization of actin cytoskeleton in the two concentrations used. Also, Hcy-induced hypophosphorylation of vimentin and glial fibrillary acidic protein (GFAP) and this effect was prevented by DL-AP5, MCPG, and CNQX. In conclusion, our results show that Hcy target the cytoskeleton of C6 cells probably by excitoxicity and/or oxidative stress mechanisms. Therefore, we could propose that the dynamic restructuring of the actin cytoskeleton of glial cells might contribute to the response to the injury provoked by elevated Hcy levels in brain.

    Topics: Actins; Animals; Antioxidants; Ascorbic Acid; Cell Line; Chromans; Cytoskeleton; Excitatory Amino Acid Agonists; Folic Acid; Glial Fibrillary Acidic Protein; Homocysteine; Intermediate Filaments; Neuroglia; Phosphorylation; Rats; Vimentin; Vitamin B Complex

2010
Antioxidant and cytotoxic activities of Aphanes arvensis extracts.
    Plant foods for human nutrition (Dordrecht, Netherlands), 2010, Volume: 65, Issue:1

    The objectives of this study were to examine the free radical scavenging activity and the protective effects against macromolecular oxidation as well as the cytotoxic activity of Aphanes arvensis aqueous and methanolic extracts. Free radical scavenging activity was determined by DPPH method. The methanolic extract showed a scavenging activity nearly equivalent to Trolox and Vitamin C and has an IC(50) value of 4.54 microg/mL. Total antioxidant capacity was determined by CUPRAC method. The antioxidant capacity of aqueous and methanolic extract was 0.792 and 1.550 mmol TE/g DWE, respectively. The protective effect of A. arvensis extracts against lipid peroxidation was evaluated using a liposome oxidation system. The methanolic extract was more active than the aqueous extract. The aqueous extract possessed protective effect against protein oxidation in a dose dependent manner. Both extracts showed inhibitory effect on DNA oxidation as measured by plasmid relaxation assay. Results presented here indicate that A. arvensis possess strong antioxidant activity and protective effects with very little cytotoxic effect, and they can therefore be used as a natural additive in food, cosmetic and pharmaceutical industries.

    Topics: Antioxidants; Ascorbic Acid; Chromans; Cytotoxins; DNA; DNA Damage; Dose-Response Relationship, Drug; Free Radical Scavengers; Inhibitory Concentration 50; Lipid Peroxidation; Liposomes; Oxidation-Reduction; Plant Extracts; Protein Carbonylation; Rosaceae

2010
Oxygen radical absorbance capacity of phycocyanin and phycocyanobilin from the food supplement Aphanizomenon flos-aquae.
    Journal of medicinal food, 2010, Volume: 13, Issue:1

    The oxygen radical absorbance capacity (ORAC) assay has been widely used to quantify peroxyl radical scavenging capacity of pure antioxidant compounds and antioxidant plant/food extracts. However, it has never been applied to natural compounds derived from microalgae-based dietary supplements, namely, phycocyanin (PC) and phycocyanobilin (PCB), for which a strong radical scavenger activity has been documented. In this article, we applied the ORAC method to investigate the capacity of PC and PCB purified from the edible microalga Aphanizomenon flos-aquae to directly quench peroxyl radicals in comparison to well-known antioxidants molecules such as Trolox, ascorbic acid, and reduced glutathione. As a result, PCB was found to have the highest ORAC value (22.18 micromol of Trolox/micromol of compound), comparable to that of PC (20.33 micromol of Trolox/micromol of compound), hence confirming that PCB is mostly responsible for the scavenger activity of PC and making the protein a possible source of the antioxidant in vivo. Our data further corroborate the use of these natural compounds from A. flos-aquae as dietary antioxidant supplements in the treatment of clinical conditions related to oxidative stress.

    Topics: Antioxidants; Aphanizomenon; Ascorbic Acid; Chromans; Dietary Supplements; Glutathione; Oxidative Stress; Peroxides; Phycobilins; Phycocyanin; Plant Preparations; Reactive Oxygen Species

2010
Role of oxidative stress and MAPK signaling in reference moist smokeless tobacco-induced HOK-16B cell death.
    Toxicology letters, 2010, May-19, Volume: 195, Issue:1

    The use of smokeless tobacco products is often associated with an oral injury at the site of repeated use. To further our understanding of this injury process, the effect of reference moist smokeless tobacco extract (STE) on cell death, oxidative stress, and MAPK signaling in a human oral keratinocyte cell line, HOK-16B, was investigated. STE caused dose-dependent cell death and reactive oxygen species (ROS) production within 30 min to 3h of exposure. This same insult enhanced the activity of ERK1/2, JNK1/2, p38 MAPK and ASK1, an upstream activator of JNK1/2 and p38 MAPK. Inhibition of JNK1/2 and to a lesser extent p38 MAPK, but not ERK1/2, suppressed STE-induced cell death. Pretreatment with antioxidants and an iron chelator, deferoxamine suppressed ROS production, ASK1, JNK1/2 and p38 MAPK activation, and reduced cell death after STE exposure. Interestingly, extracellular free iron levels in STE (29.4+/-0.5 microM) were significantly elevated as compared with cell culture medium (4.9+/-0.6 microM) and the addition of extracellular free iron (14, 30 or 70 microM) to HOK-16B cultures (without STE) caused dose-dependent cell death after 3h. Thus, acute exposure to STE leads to HOK-16B cell death in part through oxidative stress via activation of ASK1 and the JNK1/2 and p38 MAPK pathways.

    Topics: Ascorbic Acid; Cell Death; Cell Line; Chromans; Deferoxamine; Dose-Response Relationship, Drug; Gene Expression Regulation; Humans; Keratinocytes; MAP Kinase Signaling System; Mitogen-Activated Protein Kinase Kinases; Mitogen-Activated Protein Kinases; Oxidative Stress; Time Factors; Tobacco, Smokeless

2010
Determination of antioxidant constituents in cactus pear fruits.
    Plant foods for human nutrition (Dordrecht, Netherlands), 2010, Volume: 65, Issue:3

    An analytical study was carried out on the presence of antioxidant constituents and the in vitro antioxidant capacity in the extracts of three species of Spanish red-skinned cactus pear fruits (Opuntia ficus-indica, Opuntia undulata and Opuntia stricta). The cactus pear fruit extracts were analyzed for determined constituents: ascorbic acid, flavonoids (quercetin, isorhamnetin, myricetin, kaempferol and luteolin), betalains, taurine, total carotenoids and total phenolics. The antioxidant capacity was assessed by means of two different methods: the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (Trolox equivalent antioxidant capacity) method and the 2,2-diphenyl-1-picrylhydrazyl radical method. Opuntia ficus-indica fruit extract had the strongest antioxidant capacity and taurine content. O. stricta fruits were the richest in ascorbic acid and total phenolics, whereas O. undulata fruits showed the highest carotenoid content. Quercetin and isorhamnetin were the main flavonoids detected. This study provides basic information on the presence of bioactive compounds and antioxidant capacity in extracts of cactus pear fruits, in order to consider these extracts as ingredient for the production of health-promoting food.

    Topics: Antioxidants; Ascorbic Acid; Biphenyl Compounds; Carotenoids; Chromans; Flavonoids; Flavonols; Fruit; Opuntia; Phenols; Picrates; Plant Preparations; Polyphenols; Quercetin; Taurine

2010
Evaluation of selected food supplements containing antioxidants.
    Roczniki Panstwowego Zakladu Higieny, 2010, Volume: 61, Issue:2

    Seven commercial food supplements present on the Polish market were evaluated for their in vitro antioxidant capacity. The selected products were in the form of hard gelatin capsules. They contained the extracts from chokeberry, cranberry, blueberry and green tea. The mixture of vitamins and minerals as well as the product containing vitamin C in substantial dose were included into comparison. The products were examined using three methods in order to evaluate their antioxidant capacity: electron paramagnetic resonance (EPR), Trolox equivalent antioxidant capacity (TEAC), oxygen radical absorbing antioxidant capacity (ORAC) assays. Total polyphenolic content was determined according to Folin-Ciocalteu method The results were calculated per capsule. All studied preparations showed antioxidant properties and may provide substantial antioxidant protection. The in vitro antioxidant capacity varied considerably and was associated with the content of polyphenols in the capsule. The supplement with 250 mg of green tea extract was the most potent antioxidant in all assays. Nevertheless it must be remembered that the amounts of extracts were different in encapsulated products. As the quality of extracts and their properties are miscellaneous, there is a need to standardize dietary antioxidant supplements with respect to their antioxidant capacity if effective doses are to be recommended.

    Topics: Antioxidants; Ascorbic Acid; Chromans; Dietary Supplements; Electron Spin Resonance Spectroscopy; Food Analysis; Humans; Phenols; Plant Extracts; Poland; Quality Control; Reference Standards; Vitamin E; Vitamins

2010
Minimization of free radical damage by metal catalysis of multivitamin/multimineral supplements.
    Nutrition journal, 2010, Nov-23, Volume: 9

    Multivitamin/multimineral complexes are the most common dietary supplements. Unlike minerals in foods that are incorporated in bioorganic structures, minerals in dietary supplements are typically in an inorganic form. These minerals can catalyze the generation of free radicals, thereby oxidizing antioxidants during digestion. Here we examine the ability of a matrix consisting of an amino acid and non-digestible oligosaccharide (AAOS) to blunt metal-catalyzed oxidations. Monitoring of ascorbate radical generated by copper shows that ascorbate is oxidized more slowly with the AAOS matrix than with copper sulfate. Measurement of the rate of oxidation of ascorbic acid and Trolox® by catalytic metals confirmed the ability of AAOS to slow these oxidations. Similar results were observed with iron-catalyzed formation of hydroxyl radicals. When compared to traditional forms of minerals used in supplements, we conclude that the oxidative loss of antioxidants in solution at physiological pH is much slower when AAOS is present.

    Topics: Amino Acids; Antioxidants; Ascorbic Acid; Catalysis; Chelating Agents; Chromans; Copper; Dietary Supplements; Free Radical Scavengers; Free Radicals; Iron; Kinetics; Minerals; Oligosaccharides; Oxidation-Reduction; Oxidative Stress; Pharmaceutic Aids; Vitamins

2010
Inhibitory effect of antioxidants on hydroxyl radical generation from methylguanidine: an ESR study.
    Neurochemical research, 2009, Volume: 34, Issue:4

    Methylguanidine (MG) is known as not only a nephrotoxin but also as a neurotoxine. We have previously showed that MG itself generates hydroxyl radicals (*OH) in an in vitro study. In this study, we examined the inhibitory effects of ascorbate, EPC-K(1) (alpha-tocopheryl-L-ascorbate-2-O-phosphate diester), Trolox (water-soluble vitamin E analogue), and glutathione (GSH) on *OH generation from MG using an electron spin resonance (ESR) spectrometry with spin trap 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). It was found that these compounds have potent inhibitory effect on *OH generation from MG in the order of ascorbate > GSH > EPC-K(1) > Trolox.

    Topics: Antioxidants; Ascorbic Acid; Chromans; Cyclic N-Oxides; Electron Spin Resonance Spectroscopy; Glutathione; Hydroxyl Radical; Methylguanidine; Spin Trapping; Vitamin E

2009
Antioxidant activity assay based on laccase-generated radicals.
    Analytical and bioanalytical chemistry, 2009, Volume: 393, Issue:2

    A novel antioxidant activity assay was developed using laccase-oxidized phenolics. In a three-step approach, phenolic compounds were first oxidized by laccase. Laccase was then inhibited using 80% (v/v) methanol which also stabilized the oxidized phenolics which were then used to measure antioxidant activities of ascorbic acid and Trolox. From a number of laccase-oxidized phenolics screened for potential use in the measurement of antioxidant activities, syringaldazine emerged the best, giving results comparable to the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, which is currently used in conventional methods. Like DPPH radicals, two moles of stoichiometric oxidized syringaldazine were reduced by one mole of either ascorbic acid or Trolox. For the first time we show that antioxidant activity can be correlated to oxygen consumption by laccase. Reduction of one molecule of oxygen corresponded to oxidation of four molecules of syringaldazine which in turn is reduced by two molecules of Trolox or ascorbic acid. This study therefore demonstrates the great potential of using laccase-oxidized syringaldazine for the measurement of antioxidant activity.

    Topics: Antioxidants; Ascorbic Acid; Biological Assay; Biphenyl Compounds; Chromans; Free Radicals; Laccase; Methanol; Oxidation-Reduction; Oxygen; Phenols; Picrates; Spectrophotometry, Ultraviolet; Time Factors

2009
Antioxidant status associated with inflammation in sarcoidosis: a potential role for antioxidants.
    Respiratory medicine, 2009, Volume: 103, Issue:3

    Enhanced production of reactive oxygen species (ROS), capable of reducing endogenous defense levels and enhancing inflammation, is suggested to play a role in sarcoidosis. Antioxidant supplementation might offer protection against such ROS-mediated damage. A promising candidate for antioxidant supplementation is the flavonoid quercetin.. To determine the antioxidant and inflammatory status in sarcoidosis. Furthermore, the potential of quercetin to mitigate the occurring inflammation will be assessed.. Non-smoking sarcoidosis patients and healthy controls matched for age, gender and dietary behavior were enrolled (NCT-00512967). Measurements included assessment of total plasma antioxidant capacity, vitamin C, uric acid, glutathione, basal and LPS-induced levels of tumor necrosis factor alpha (TNFalpha), interleukin (IL)-8 and -10 as well as the effect of quercetin on these levels.. Compared to their controls, the sarcoidosis patients displayed significantly lower total plasma antioxidant capacity, decreased levels of vitamin C, uric acid and glutathione and increased levels of basal TNFalpha and IL-8. Quercetin significantly decreased ex vivo LPS-induced TNFalpha- and IL-8 production in a concentration-dependent manner in both groups. Interestingly, this quercetin effect was more pronounced in sarcoidosis patients.. The endogenous antioxidant defense was significantly reduced in sarcoidosis, indicating that oxidative stress underlies the pathology of this disease. Furthermore, the inflammatory status was significantly enhanced in sarcoidosis. Finally, our results regarding the effect of quercetin on cytokine production imply that sarcoidosis patients might benefit from antioxidant supplementation not only by empowering the relatively low protection against ROS but also by reducing inflammation.

    Topics: Adult; Antioxidants; Ascorbic Acid; Biomarkers; Case-Control Studies; Chromans; Dose-Response Relationship, Drug; Female; Flavonoids; Glutathione; Humans; Interleukin-10; Interleukin-8; Lipopolysaccharides; Lung; Male; Middle Aged; Oxidative Stress; Quercetin; Sarcoidosis; Statistics, Nonparametric; Tumor Necrosis Factor-alpha; Uric Acid

2009
A new redox-dependent mechanism of MMP-1 activity control comprising reduced low-molecular-weight thiols and oxidizing radicals.
    Journal of molecular medicine (Berlin, Germany), 2009, Volume: 87, Issue:3

    Matrix metalloproteinases (MMPs), a family of zinc-dependent proteinases, participate in remodeling and degradation of the extracellular matrix proteins. The activity of MMPs is thought to be predominately posttranslationally regulated via proteolytic activation of precursor zymogens or via their naturally occurring endogenous inhibitors. Here, using recombinant MMP-1, we investigated new redox-dependent mechanisms of proteinase activity regulation by low-molecular-weight thiols. We find that glutathione (GSH), cysteine, homocysteine, and N-acetylcysteine at physiological concentrations competitively reduce MMP-1 activity up to 75% with an efficiency of cysteine > or = GSH > homocysteine > N-acetylcysteine. In contrast, S-derivatized thiols completely lack this inhibitory activity. Interestingly, the competitive GSH-mediated inhibition of MMP-1-activity can be fully reversed abrogated by oxidizing radicals like (*)NO(2) or Trolox radicals, here generated by UVA irradiation of nitrite or Trolox, two relevant agents in human skin physiology. This redox-dependent reactivation of the inactive GSH-MMP-1-complex comprises GSH oxidation and is significantly inhibited in the presence of ascorbic acid, an effective (*)NO(2) and Trolox radical scavenger. We here offer a new concept of redox-sensitive control of MMP-1 activity based on the inhibitory effect of reduced thiols and reactivation by a mechanism comprising derivatization or oxidation of the MMP-1-bound inhibitory-acting thiol.

    Topics: Antioxidants; Ascorbic Acid; Blotting, Western; Catalysis; Cells, Cultured; Chromans; Electron Spin Resonance Spectroscopy; Fluorescence; Free Radicals; Glutathione; Glutathione Disulfide; Humans; Kinetics; Matrix Metalloproteinase 1; Matrix Metalloproteinase Inhibitors; Molecular Structure; Molecular Weight; Nitrites; Oxidation-Reduction; Peptides; Recombinant Proteins; Substrate Specificity; Sulfhydryl Compounds; Ultraviolet Rays

2009
Comparison of bee products based on assays of antioxidant capacities.
    BMC complementary and alternative medicine, 2009, Feb-26, Volume: 9

    Bee products (including propolis, royal jelly, and bee pollen) are popular, traditional health foods. We compared antioxidant effects among water and ethanol extracts of Brazilian green propolis (WEP or EEP), its main constituents, water-soluble royal jelly (RJ), and an ethanol extract of bee pollen.. The hydrogen peroxide (H2O2)-, superoxide anion (O2.-)-, and hydroxyl radical (HO.)- scavenging capacities of bee products were measured using antioxidant capacity assays that employed the reactive oxygen species (ROS)-sensitive probe 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCFDA) or aminophenyl fluorescein (APF).. The rank order of antioxidant potencies was as follows: WEP > EEP > pollen, but neither RJ nor 10-hydroxy-2-decenoic acid (10-HDA) had any effects. Concerning the main constituents of WEP, the rank order of antioxidant effects was: caffeic acid > artepillin C > drupanin, but neither baccharin nor coumaric acid had any effects. The scavenging effects of caffeic acid were as powerful as those of trolox, but stronger than those of N-acetyl cysteine (NAC) or vitamin C.. On the basis of the present assays, propolis is the most powerful antioxidant of all the bee product examined, and its effect may be partly due to the various caffeic acids it contains. Pollen, too, exhibited strong antioxidant effects.

    Topics: Acetylcysteine; Animals; Ascorbic Acid; Bees; Caffeic Acids; Chromans; Cinnamates; Coumaric Acids; Fatty Acids; Fatty Acids, Monounsaturated; Free Radical Scavengers; Phenylpropionates; Plant Extracts; Pollen; Propolis; Quinic Acid; Trichothecenes

2009
[The comparison of alpha-lipoic acid, melatonin, vitamin C and trolox effectiveness in decreasing DNA stand brakes and increasing plasma antioxidant power].
    Polski merkuriusz lekarski : organ Polskiego Towarzystwa Lekarskiego, 2009, Volume: 27, Issue:157

    Human plasma contains different antioxidants which may be important in the maintenance of an antioxidant status. The aim of this study was to compare the effect of exogenous antioxidants (vitamin C, trolox, alpha-lipoic acid and melatonin) on DNA standard brakes level and antioxidant activity of plasma.. Blood was obtain from 24 healthy volunteers from 19-23 of age and centrifuge. Vitamin C (vit C), trolox (water soluble vitamin E), lipoic acid (LA) or melatonin (MEL) were added separately to the plasma right before assay. Final concentration of each antioxidant in plasma was 1 mmol. Antioxidant properties of plasma and plasma with antioxidants were evaluated by three methods: level of DNA damages, ferric reducing ability of plasma (FRAP), scavenging of DPPH (2,2'-diphenyl-1-picrylhydrazyl) radical.. We showed that vit C and trolox have strong ability to reduce bivalent iron (p<0.001 and p<0.001, respectively). The highest protective effect on hydroxyl radical-induced DNA damage was observed for trolox and LA (p<0.001 and p<0.01, respectively). All antioxidants possessed weak ability to scavenge DPPH radicals.. In our study trolox and vitamin C effectively diminished DNA oxidative damages and possessed the highest antioxidant properties. Melatonin and lipoic acid decreased DNA damages but did not increase plasma antioxidant ability.

    Topics: Adult; Antioxidants; Ascorbic Acid; Chromans; DNA Breaks; Humans; In Vitro Techniques; Melatonin; Reference Values; Thioctic Acid; Young Adult

2009
Intracellular glutathione regulates Andrographolide-induced cytotoxicity on hepatoma Hep3B cells.
    Redox report : communications in free radical research, 2009, Volume: 14, Issue:4

    Andrographolide (ANDRO), a diterpenoid lactone isolated from the traditional herbal plant Andrographis paniculata, was reported to induce apoptosis in hepatoma Hep3B cells in our previous study (Ji LL, Liu TY, Liu J, Chen Y, Wang ZT. Andrographolide inhibits human hepatoma-derived Hep3B cells growth through the activation of c-Jun N-terminal kinase. Planta Med 2007; 73: 1397-1401). The present investigation was carried out to observe whether cellular reduced glutathione (GSH) plays important roles in ANDRO-induced apoptosis. ANDRO initially increased intracellular GSH levels which then decreased later, while inhibition of cellular GSH synthesis by L-Buthionine-(S,R)-sulfoximine (BSO) augmented ANDRO-induced cytotoxicity and apoptosis in Hep3B cells. On the other hand, the thiol antioxidant dithiothreitol (DTT) rescued ANDRO-depleted cellular GSH, and abrogated ANDRO-induced cytotoxicity and apoptosis. Furthermore, BSO pretreatment augmented ANDRO-decreased expression of antioxidant protein thioredoxin 1 (Trx1), while DTT reversed this decrease. Further results showed that ANDRO increased the activity of the GSH-related antioxidant enzyme glutathione peroxidase (GPx) and the production of intracellular reactive oxygen species (ROS). Taken together, this study demonstrates that the intracellular redox system plays important roles in regulating the cytotoxicity of ANDRO on hepatoma Hep3B cells.

    Topics: Antioxidants; Apoptosis; Ascorbic Acid; Blotting, Western; Cell Line, Tumor; Cell Survival; Chromans; Diterpenes; Dithiothreitol; DNA Fragmentation; Glutathione; Glutathione Peroxidase; Humans; Mannitol; Reactive Oxygen Species

2009
ESR study of the singlet oxygen quenching and protective activity of Trolox on the photodecomposition of riboflavin and lumiflavin in aqueous buffer solutions.
    Journal of food science, 2009, Volume: 74, Issue:6

    Singlet oxygen quenching activity of Trolox, a water-soluble derivative of tocopherol, was studied by electron spin resonance (ESR) spectroscopy in a buffer solution (pH 7.4) containing methylene blue (MB), 2,2,6,6-tetramethyl-4-piperidone (TMPD) after light illumination for 30 min. Trolox at the concentration of 125 microM quenched 89.1% singlet oxygen in the system. Trolox showed significantly higher singlet oxygen quenching activity than ascorbic acid in the buffer solution (P < 0.05). Riboflavin in phosphate buffer solutions was degraded very fast under fluorescent light illumination. The photodegradation rate of riboflavin at pH 8.5 was significantly higher than pHs 4.5 and 6.5 (P < 0.05). Lumiflavin was also degraded under the fluorescent light illumination, but its degradation rate was much lower than that of riboflavin under the same light intensity. Unlike riboflavin, the rate of lumiflavin photodegradation was the greatest at pH 4.5 and followed by pHs 6.5 and 8.5, in a decreasing order. Trolox greatly protected the photodegradation of riboflavin and lumiflavin. The protective activities of Trolox against the photodegradation of riboflavin and lumiflavin were also pH dependent. The treatments of 5 mM Trolox in the buffer solutions of pHs 8.5 and 6.5 exhibited 56.1% and 31.7% protection of riboflavin against degradation during 120 min light illumination, respectively. The treatments of Trolox at the concentrations of 1, 3, and 5 mM in the buffer of 6.5 exhibited 14.8%, 58.4%, and 81.4% protection of lumiflavin against degradation during 24 h light illumination, respectively.

    Topics: Antioxidants; Ascorbic Acid; Chromans; Chromatography, High Pressure Liquid; Electron Spin Resonance Spectroscopy; Flavins; Hydrogen-Ion Concentration; Light; Photolysis; Riboflavin; Singlet Oxygen; Time Factors

2009
A high-throughput reporter gene assay to prove the ability of natural compounds to modulate glutathione peroxidase, superoxide dismutase and catalase gene promoters in V79 cells.
    Free radical research, 2008, Volume: 42, Issue:8

    The aim of the study was to establish a 96-well microtiter plate-based reporter gene assay to test the influence of natural compounds on the promoter activities of rat catalase, human glutathione peroxidase and human superoxide dismutase expressed in V79 cells. Luciferase expression vectors with the promoter regions of the genes coding for the three above-mentioned enzymes were constructed and transfected into V79 cells. Thereafter the ability of sodium ascorbate, L-carnitine, catechin, epigallocatechin gallate, genistein, paraquat, quercetin, 12-O-tetradecanoylphorbol-13-acetate and Trolox to enhance the promoter activities was evaluated. Genistein, paraquat and quercetin led to a statistically significant increase in the glutathione peroxidase and superoxide dismutase gene promoter activities. None of the compounds tested enhanced the catalase gene promoter activity. The reporter gene assay described in this report is easy to perform, fast and allows one to test a high number of compounds and different concentrations of a single compound at the same time.

    Topics: Animals; Anticarcinogenic Agents; Antioxidants; Ascorbic Acid; Carcinogens; Carnitine; Catalase; Catechin; Cells, Cultured; Chromans; Cricetinae; Cricetulus; Genistein; Glutathione Peroxidase; Herbicides; Humans; Luciferases; Paraquat; Promoter Regions, Genetic; Quercetin; Rats; Superoxide Dismutase; Tetradecanoylphorbol Acetate; Vitamin B Complex

2008
Protective effect of antioxidants against sarcoplasmic reticulum (SR) oxidation by Fenton reaction, however without prevention of Ca-pump activity.
    Toxicology in vitro : an international journal published in association with BIBRA, 2008, Volume: 22, Issue:7

    The Ca(2+)-ATPase of the sarcoplasmic reticulum (SERCA) of rabbit skeletal muscle was oxidized by Fe2+/H2O2/ascorbic acid (AA), a system which generates HO(.) radicals according to the Fenton reaction: (Fe2(+)+H2O2-->HO(.)+OH(-)+Fe(3+)) under conditions similar to the pathological state of inflammation. Under these conditions, when hydroxyl-radicals and/or ferryl-radicals are generated, a 50% decrease of the SERCA activity was observed, a significant decrease of SH groups and an increase of protein carbonyl groups and lipid peroxidation were identified. Two new bands, time dependent in density, appeared in the SERCA protein electrophoresis after incubation with the Fenton system (at approximately 50 and 75kDa), probably due to structural changes as supported also by trypsin digestion. Immunoblotting of DNPH derivatized protein bound carbonyls detected a time dependent increase after incubation of SERCA with the Fenton system. Trolox and the pyridoindole stobadine (50microM) protected SR against oxidation induced via the Fenton system by preventing SH group oxidation and lipid peroxidation. Pycnogenol((R)) and EGb761 (40microg/ml) protected SERCA in addition against protein bound carbonyl formation. In spite of the antioxidant effects, trolox and stobadine were not able to prevent a decrease in the SERCA Ca(2+)-ATPase activity. Pycnogenol and EGb761 even enhanced the decrease of the Ca(2+)-ATPase activity induced by the Fenton system, probably by secondary oxidative reactions.

    Topics: Animals; Antioxidants; Ascorbic Acid; Carbolines; Chromans; Ferrous Compounds; Flavonoids; Ginkgo biloba; Hydrogen Peroxide; Inflammation; Lipid Peroxidation; Muscle, Skeletal; Oxidation-Reduction; Plant Extracts; Protein Carbonylation; Rabbits; Sarcoplasmic Reticulum Calcium-Transporting ATPases; Swine; Time Factors

2008
Adiposity measures and oxidative stress among police officers.
    Obesity (Silver Spring, Md.), 2008, Volume: 16, Issue:11

    Our objective was to investigate associations between adiposity measures (BMI, waist circumference, waist-to-hip ratio, waist-to-height ratio, and abdominal height) and biomarkers of oxidative stress (glutathione (GSH), GSH peroxidase (GSH-Px), vitamin C, thiobarbituric acid reactive substances (TBARS), and trolox equivalent antioxidant capacity (TEAC)) among police officers. This cross-sectional study included randomly selected police officers (43 policewomen; 67 policemen) from Buffalo, New York. Adiposity measures were performed using standardized methods. Biomarkers were measured on fasting blood specimens. An oxidative stress score (OSS) was created as a composite of the biomarkers. ANOVAs were used to compare mean levels of biomarkers across tertiles of the adiposity measures. Officers were 26- to 61-years old. GSH was inversely associated with waist circumference (trend P = 0.030) and waist-to-hip ratio (trend P = 0.026). GSH-Px was inversely associated with BMI (trend P = 0.004) and with waist-to-height ratio (trend P = 0.017). No associations were observed for TEAC, TBARS, or OSS with any adiposity measure. Significant interactions were observed by physical activity status for GSH with waist circumference and waist-to-hip ratio and for vitamin C with waist circumference, waist-to-hip and waist-to-height ratios. The above associations were inversely related only among officers who reported engaging in physical activity. Inverse associations were observed for BMI and waist circumference with GSH, but only among women; the interaction with gender was significant. Larger indices of adiposity were associated with increased levels of oxidative stress and decreased levels of antioxidant defense.

    Topics: Adiposity; Adult; Ascorbic Acid; Biomarkers; Body Mass Index; Chromans; Cross-Sectional Studies; Female; Glutathione; Glutathione Peroxidase; Humans; Male; Middle Aged; Oxidative Stress; Police; Thiobarbituric Acid Reactive Substances; Waist-Hip Ratio

2008
Quenching mechanisms and kinetics of Trolox and ascorbic acid on the riboflavin-photosensitized oxidation of tryptophan and tyrosine.
    Journal of agricultural and food chemistry, 2008, Nov-26, Volume: 56, Issue:22

    The effects of 0, 0.3, 0.6, and 0.9 mM Trolox and ascorbic acid on the singlet oxygen oxidation of tryptophan and tyrosine containing 25 ppm of riboflavin were determined by measuring tryptophan and tyrosine concentration by high-performance liquid chromatography analysis. The samples were stored in the a 1000 lx light storage box for 4 h at 30 degrees C. As the concentration of Trolox and ascorbic acid increased, the degradation of tryptophan and tyrosine decreased significantly at p < 0.05. Trolox reduced tryptophan and tyrosine degradation by quenching both singlet oxygen and excited triplet riboflavin, whereas ascorbic acid quenched singlet oxygen only. The total singlet oxygen quenchings of Trolox in the presence of tryptophan and tyrosine were 1.55 x 10(7) and 1.32 x 10(7) M(-1) s(-1), respectively. The total singlet oxygen quenchings of ascorbic acid in the presence of tryptophan and tyrosine were 1.16 x 10(7) and 1.10 x 10(7) M(-1) s(-1), respectively. Trolox was more effective than ascorbic acid in preventing the degradation of tryptophan and tyrosine.

    Topics: Ascorbic Acid; Chromans; Kinetics; Oxidation-Reduction; Photochemistry; Riboflavin; Singlet Oxygen; Tryptophan; Tyrosine

2008
The antioxidant and pro-oxidant activity of vitamin C and trolox in vitro: a comparative study.
    Journal of applied toxicology : JAT, 2008, Volume: 28, Issue:2

    The antioxidant and pro-oxidant properties of ascorbic acid (vitamin C) and the water-soluble analogue of alpha-tocopherol (trolox) were compared. Trolox has advantages over alpha-tocopherol, the latter being only lipid-soluble due to the presence of a carboxyl group in lieu of a phytol chain which imparts trolox with water solubility. Trolox is used as a standard antioxidant in biochemical studies against which the antioxidant capacity of compounds is compared. Although ascorbic acid and tocopherols possess strong antioxidant properties, they might also exhibit pro-oxidant properties in the presence of free transition metals. Thus, reactions detailed in this study were performed in the presence of Cr(VI) in an effort to investigate the potential of ascorbic acid and trolox to generate hydroxyl radicals in a Fenton-like reaction. Results obtained were derived from reactions containing the same concentration of ascorbic acid and trolox under identical experimental conditions. Hydroxyl radical formation was observed in the reaction of Cr(VI) with ascorbic acid resulting from ascorbic acid auto-oxidation and H2O2 formation. Hydroxyl radical formation was only detected in the reaction mixture containing Cr(VI) and trolox following the addition of H2O2.

    Topics: Antioxidants; Ascorbic Acid; Chromans; Chromium; DNA Damage; DNA, Superhelical; Hydrogen Peroxide; Hydroxyl Radical; Oxidants; Oxidation-Reduction; Plasmids

2008
Glial cell line-derived neurotrophic factor and antioxidants preserve the electrical responsiveness of the spiral ganglion neurons after experimentally induced deafness.
    Neurobiology of disease, 2008, Volume: 29, Issue:1

    Cochlear implant surgery is currently the therapy of choice for profoundly deaf patients. However, the functionality of cochlear implants depends on the integrity of the auditory spiral ganglion neurons. This study assesses the combined efficacy of two classes of agents found effective in preventing degeneration of the auditory nerve following deafness, neurotrophic factors, and antioxidants. Guinea pigs were deafened and treated for 4 weeks with either local administration of GDNF or a combination of GDNF and systemic injections of the antioxidants ascorbic acid and Trolox. The density of surviving spiral ganglion cells was significantly enhanced and the thresholds for eliciting an electrically evoked brain stem response were significantly reduced in GDNF treated animals compared to deafened-untreated. The addition of antioxidants significantly enhanced the evoked responsiveness over that observed with GDNF alone. The results suggest multiple sites of intervention in the rescue of these cells from deafferentation-induced cell death.

    Topics: Acoustic Stimulation; Animals; Antioxidants; Ascorbic Acid; Auditory Threshold; Chromans; Deafness; Disease Models, Animal; Dose-Response Relationship, Radiation; Drug Administration Routes; Electric Stimulation; Evoked Potentials, Auditory, Brain Stem; Glial Cell Line-Derived Neurotrophic Factor; Guinea Pigs; Neurons; Spiral Ganglion; Statistics, Nonparametric

2008
Microplate-compatible biamperometry array for parallel 48-channel amperometric or coulometric measurements.
    Analytical chemistry, 2008, Apr-15, Volume: 80, Issue:8

    We report a new reusable electrochemical array for parallel biamperometric measurements that has been designed for use with standard microplates. The 48-channel array uses half of the available 96 wells and has 48 pairs of Pt wire electrodes. Applications to the quantitation of a variety of oxidizable species, including acetaminophen, ascorbic acid, hydroquinone, trolox, and uric acid, are demonstrated in assays that use potassium ferricyanide as an oxidant to produce a mixture of ferri- and ferrocyanide. Hydrogen peroxide quantitation is also demonstrated, based on an assay in which ferrocyanide is oxidized, again to produce a mixture of ferri- and ferrocyanide. Detection limits (signal-to-noise ratio (S/N) = 3) in these assays range from 1 (acetaminophen, R2 = 0.994) to 8 microM (ascorbic acid, R2 = 0.967), and linearity was observed to analyte concentrations of at least 100 microM. We also demonstrate the application of the biamperometric array to enzymatic assays, using the glucose oxidase reaction as an example; following a 20 min enzyme reaction time, a detection limit of 0.1 mM glucose was obtained. These results indicate that applications to other oxidase-based assays are feasible in this high-throughput format. The new electrochemical array employs standard, inexpensive microplates, and the biamperometric measurements are simple, precise, and rapid, requiring only 2 min for 48 parallel measurements.

    Topics: Acetaminophen; Ascorbic Acid; Calibration; Chromans; Electrochemistry; Electrodes; Ferrocyanides; Glucose; Glucose Oxidase; Hydrogen Peroxide; Hydroquinones; Microchemistry; Oxidation-Reduction; Platinum; Uric Acid

2008
Effects of antioxidants on auditory nerve function and survival in deafened guinea pigs.
    Neurobiology of disease, 2007, Volume: 25, Issue:2

    Based on in vitro studies, it is hypothesized that neurotrophic factor deprivation following deafferentation elicits an oxidative state change in the deafferented neuron and the formation of free radicals that then signal cell death pathways. This pathway to cell death was tested in vivo by assessing the efficacy of antioxidants (AOs) to prevent degeneration of deafferented CNVIII spiral ganglion cells (SGCs) in deafened guinea pigs. Following destruction of sensory cells, guinea pigs were treated immediately with Trolox (a water soluble vitamin E analogue)+ascorbic acid (vitamin C) administered either locally, directly in the inner ear, or systemically. Electrical auditory brainstem response (EABR) thresholds were recorded to assess nerve function and showed a large increase following deafness. In treated animals EABR thresholds decreased and surviving SGCs were increased significantly compared to untreated animals. These results indicate that a change in oxidative state following deafferentation plays a role in nerve cell death and antioxidant therapy may rescue SGCs from deafferentation-induced degeneration.

    Topics: Animals; Antioxidants; Ascorbic Acid; Auditory Threshold; Cell Survival; Chromans; Cochlear Nerve; Deafness; Denervation; Evoked Potentials, Auditory, Brain Stem; Guinea Pigs; Hair Cells, Auditory; Hearing Loss, Sensorineural; Male; Neurons, Afferent; Neuroprotective Agents; Oxidative Stress; Spiral Ganglion; Treatment Outcome

2007
Inhibition of Neutral red photolysis with different antioxidants.
    Bosnian journal of basic medical sciences, 2007, Volume: 7, Issue:1

    Neutral red is a dye the azine structure which has been used as an acido-base indicator and a dye in histochemistry. In 1960 Goldhaber introduced Neutral red into the medium of resorbing bone cultures to localize the osteoclast in the living cultures. Using time-lapse microcinematography in order to follow the osteoclasts, he reported excellent contrast could be obtained with Neutral red due to the avidity of osteoclasts for this dye. Unfortunately, however, the photodynamic effect resulting from subsequent exposure of these cultures to light precluded this approach, and again in 1963. it was observed that the death of the osteoclasts was probably due to a photodynamic effect related to the dye in the cell, the presence of oxygen and the frequent exposure of light by our time-lapse photography. VIS and UV irradiation induced photolysis of Neutral red, and from Neutral red cation produced with photons a Neutral red radical. This Neutral red radical can be inhibited with action of an antioxidant, such as melatonin, glutathione, ascorbic acid, E vitamin, etc. We developed an assay with Neutral red photolysis which utilizes a VIS and UV irradiation technique for quantification the inhibition of photolysis with action of an antioxidant. In this method Neutral red acts double, as a free radical generator and as a photosensitizer.

    Topics: Antioxidants; Ascorbic Acid; Chromans; Glutathione; Melatonin; Neutral Red; Photolysis; Phototherapy; Spectrophotometry

2007
The impact of nitrite and antioxidants on ultraviolet-A-induced cell death of human skin fibroblasts.
    Free radical biology & medicine, 2007, Sep-01, Volume: 43, Issue:5

    Nitrite (NO(2)(-)) occurs ubiquitously in biological fluids such as blood and sweat. Ultraviolet A-induced nitric oxide formation via decomposition of cutaneous nitrite, accompanied by the production of reactive oxygen (ROS) or nitrogen species (RNS), represents an important source for NO in human skin physiology. Examining the impact of nitrite and the antioxidants glutathione (GSH), Trolox (TRL), and ascorbic acid (ASC) on UVA-induced toxicity of human skin fibroblasts (FB) we found that NO(2)(-) concentration-dependently enhances the susceptibility of FB to the toxic effects of UVA by a mechanism comprising enhanced induction of lipid peroxidation. While ASC completely protects FB cultures from UVA/NO(2)(-)-induced cell damage, GSH or TRL excessively enhances UVA/NO(2)(-)-induced cell death by a mechanism comprising nitrite concentration-dependent TRL radical formation or GSH-derived oxidative stress. Simultaneously, in the presence of GSH or TRL the mode of UVA/NO(2)(-)-induced cell death changes from apoptosis to necrosis. In summary, during photodecomposition of nitrite, ROS or RNS formation may act as strong toxic insults. Although inhibition of oxidative stress by NO and other antioxidants represents a successful strategy for protection from UVA/NO(2)(-)-induced injuries, GSH and TRL may nitrite-dependently aggravate the injurious impact by TRL or GSH radical formation, respectively.

    Topics: Antioxidants; Ascorbic Acid; Cell Death; Cells, Cultured; Chromans; Cytoprotection; Fibroblasts; Free Radicals; Glutathione; Humans; Lipid Peroxidation; Nitric Oxide; Nitrites; Skin; Ultraviolet Rays

2007
Mechanisms of root canal sealers cytotoxicity on osteoblastic cell line MC3T3-E1.
    Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics, 2007, Volume: 104, Issue:5

    The cytotoxic mechanisms of root canal sealers (Sealapex, AH26, and N2 Universal) were studied in vitro with MC3T3-E1 osteoblastic cells.. MC3T3-E1 cells were cotreated with root canal sealers and antioxidants, and concentrations of intracellular glutathione (GSH) and reactive oxygen species (ROS) were measured. DNA fragmentation was observed after treatment with the sealers.. N-Acetylcysteine (NAC) prevented N2 Universal- and AH26-induced cytotoxicities. However, ascorbic acid and Trolox did not affect the cytotoxicity of the sealers. N2 Universal and AH26 significantly decreased the GSH pool within a 3-hour treatment period. Unlike GSH levels, the ROS levels were not altered by the sealers. Cytotoxicity of Sealapex was not affected by NAC, and there were no changes of GSH/glutathione disulfide levels in cells treated with Sealapex.. Cytotoxicities of N2 Universal and AH26 are caused by an intracellular GSH depletion without a burst of ROS. Sealapex may cause cytotoxicity in a way different from N2 Universal and AH26.

    Topics: Animals; Antioxidants; Ascorbic Acid; Cattle; Cell Line; Chromans; Cystine; DNA Fragmentation; Glutathione; Mice; Osteoblasts; Reactive Oxygen Species; Root Canal Filling Materials

2007
Pro-apoptotic effects of 1'-acetoxychavicol acetate in human breast carcinoma cells.
    Toxicology letters, 2007, Sep-28, Volume: 173, Issue:3

    The tropical ginger compound, 1'-acetoxychavicol acetate (ACA) possesses cancer chemopreventive properties in several models but its effects on breast cancer have not been fully evaluated. In this study, the effects of ACA on human breast carcinoma-derived MCF-7 and MDA-MB-231 cell viability were assessed using trypan blue exclusion analysis. ACA significantly decreased cell viability in a time- and dose-dependent manner, with effective concentrations 10-50 microM. Apoptosis was confirmed by morphological examination of cells through light microscopy, 4,6-diamidino-2-phenylindole dihydrochloride staining, and annexin V/Alexa Fluor 488 staining visualized using flow cytometry. ACA also increased protein expression of the activated form of caspase-3 in MDA-MB-231 cells. Addition of antioxidants N-acetylcysteine, ascorbic acid, or trolox prevented the loss of viability caused by ACA using trypan blue uptake as a marker. These results suggest ACA may have potential anticancer effects against breast carcinoma cells by inducing apoptosis.

    Topics: Acetylcysteine; Antineoplastic Agents, Phytogenic; Antioxidants; Apoptosis; Ascorbic Acid; Benzyl Alcohols; Breast Neoplasms; Caspase 3; Cell Cycle; Cell Line, Tumor; Cell Survival; Chromans; Dose-Response Relationship, Drug; Enzyme Induction; Female; Humans; Terpenes; Time Factors

2007
Sequential injection analysis with chemiluminescence detection for the antioxidative activity against singlet oxygen.
    Analytical sciences : the international journal of the Japan Society for Analytical Chemistry, 2006, Volume: 22, Issue:1

    A sequential injection analysis (SIA) with chemiluminescence (CL) detection was developed for the measurement of antioxidative activity against singlet oxygen ((1)O2). Lactoperoxidase-hydrogen peroxide-bromide ion system was used for the generation of (1)O2. When a 100 mM sodium acetate buffer (pH 4.5) was used as a carrier solution, the SIA-CL system could be optimized with respect to the flow-rate of the carrier, concentration of reagents and their aspiration order. The antioxidative activity was expressed as an attenuation of luminol CL due to the quenching of (1)O2 by an antioxidant. The relative standard deviations of antioxidative activity (n=3) against (1)O2 for within- and between-day analyses were < or = 1.6% (20 microM Trolox). The system was successfully applied to the assay of antioxidative activities of various antioxidants including vitamin supplements at a rate of 10 samples within 15 min. The proposed SIA-CL system was rapid and reproducible with minimum consumption of the sample and of reagents, and thus was useful for the screening of compounds possessing antioxidative activity against (1)O2.

    Topics: Antioxidants; Ascorbic Acid; Bromides; Chromans; Flow Injection Analysis; Hydrogen Peroxide; Lactoperoxidase; Luminescent Measurements; Luminol; Sensitivity and Specificity; Singlet Oxygen; Sodium Azide; Vitamin E

2006
Fibrinogen is a co-antioxidant that supplements the vitamin E analog trolox in a model system.
    Free radical research, 2006, Volume: 40, Issue:3

    It appears that the atherosclerotic plaque is a prooxidant environment where some molecules that are normally antioxidants, including vitamins C and E, may act as prooxidants that contribute to atherosclerosis by oxidizing LDL. Some molecules can act as co-antioxidants to eliminate this prooxidant effect by recycling or other mechanisms of supplementation. Fibrinogen and other acute phase proteins found in the plaque are antioxidants. We hypothesized that fibrinogen can act as a co-antioxidant to supplement vitamin E thereby eliminating its oxidative effect under prooxidant conditions. We tested a model system for this hypothesis using the vitamin E analogue Trolox in a cell free system.. LDL was oxidized using 5 umol/l copper. Antioxidant conditions were achieved by adding the antioxidants immediately with LDL, while prooxidant conditions were created by adding antioxidants after a 40 min delay. Oxidation was monitored as the lag phase at 234 nm.. Under antioxidant conditions, the protective effect of fibrinogen and Trolox combined together were about equal to the sum of the anitioxidant effects of each alone (additive), while under prooxidant conditions the combined protection was 54-200% greater (synergistic). These effects were different than those of vitamin C with Trolox in that under antioxidant conditions fibrinogen and Trolox were additive while vitamin C and Trolox showed strong synergistic effects, and in that unlike vitamin C and Trolox fibrinogen showed no prooxidant tendencies under prooxidant reaction conditions.. The data indicated that fibrinogen did act as a co-antioxidant to supplement Trolox and eliminate its prooxidant effect, most probably, by directly quenching the phenoxyl radical, because unlike vitamin C, fibrinogen did not appear to recycle vitamin E. But fibrinogen may act as a universal antioxidant, since unlike Trolox and vitamin C, it showed little tendency toward becoming a prooxidant.

    Topics: Antioxidants; Ascorbic Acid; Chromans; Dietary Supplements; Drug Synergism; Fibrinogen; Humans; Kinetics; Lipoproteins, LDL; Models, Biological; Oxidants; Oxidation-Reduction

2006
Characterization of the radical-scavenging reaction of 2-O-substituted ascorbic acid derivatives, AA-2G, AA-2P, and AA-2S: a kinetic and stoichiometric study.
    Biological & pharmaceutical bulletin, 2006, Volume: 29, Issue:4

    The aim of this study was to characterize the antioxidant activity of three ascorbic acid (AA) derivatives O-substituted at the C-2 position of AA: ascorbic acid 2-glucoside (AA-2G), ascorbic acid 2-phosphate (AA-2P), and ascorbic acid 2-sulfate (AA-2S). The radical-scavenging activities of these AA derivatives and some common low molecular-weight antioxidants such as uric acid or glutathione against 1,1-diphenyl-picrylhydrazyl (DPPH) radical, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS+), or galvinoxyl radical were kinetically and stoichiometrically evaluated under pH-controlled conditions. Those AA derivatives slowly and continuously reacted with DPPH radical and ABTS+, but not with galvinoxyl radical. They effectively reacted with DPPH radical under acidic conditions and with ABTS+ under neutral conditions. In contrast, AA immediately quenched all species of radicals tested at all pH values investigated. The reactivity of Trolox, a water-soluble vitamin E analogue, was comparable to that of AA in terms of kinetics and stoichiometrics. Uric acid and glutathione exhibited long-lasting radical-scavenging activity against these radicals under certain pH conditions. The radical-scavenging profiles of AA derivatives were closer to those of uric acid and glutathione rather than to that of AA. The number of radicals scavenged by one molecule of AA derivatives, uric acid, or glutathione was equal to or greater than that by AA or Trolox under the appropriate conditions. These data suggest the potential usage of AA derivatives as radical scavengers.

    Topics: Ascorbic Acid; Benzhydryl Compounds; Benzothiazoles; Biphenyl Compounds; Chromans; Free Radical Scavengers; Glutathione; Hydrogen-Ion Concentration; Kinetics; Picrates; Sulfonic Acids; Uric Acid

2006
Evaluation of the free-radical-scavenging activity of diclofenac acid on the free-radical-induced haemolysis of human erythrocytes.
    The Journal of pharmacy and pharmacology, 2006, Volume: 58, Issue:5

    Free-radical-induced peroxidation in-vivo is regarded as the aetiology of some diseases and free-radical-scavenging drugs, also called antioxidants (AH), have been widely used to overcome oxidative stress. An in-vitro experimental method, 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH)-induced haemolysis of human erythrocytes can be applied to assess the free-radical-scavenging activity of a drug. The major objectives of this work were focused on three aspects. Firstly, introduction of the chemical kinetic deduction of free-radical-initiating reaction to AAPH-induced haemolysis of human erythrocytes, by which the number of free radicals trapped by an antioxidant, n, can be obtained after finding the quantitative relationship between the inhibition period (t(inh)) and the concentration of the antioxidant, t(inh) = (n/Ri) [AH]. Ri, the free-radical-initiating rate, was initially confirmed by using alpha-tocopherol (VE) whose n was taken as 2. Secondly, the free-radical-scavenging activity of diclofenac acid (DaH) and its sodium salt (DaNaH) was assessed. It has been found that DaH and DaNaH protect human erythrocytes against AAPH-induced haemolysis dose-dependently. In particular, the n values of DaH and DaNaH (4.96 and 3.60) were much higher than some traditional antioxidants, such as 6-hydroxyl-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox, a water-soluble structural analogue of VE, n = 0.30) and L-ascorbic acid (VC, n = 0.25), and L-ascorbyl-6-laurate (VC-12, a lipophilic structural analogue of VC, n = 1.11). Moreover, the free-radical-scavenging activity of lipophilic antioxidants is higher than the corresponding water-soluble species. Thirdly, the free-radical-scavenging activity of mixed antioxidants, VE + DaH, VC-12 + DaH, Trolox + DaNaH and VC + DaNaH, was revealed. The n value of VC, VC-12, VE and Trolox increase in the case of mixed usage with DaH and DaNaH, implying that diclofenac acid can repair the radical of these antioxidants. Thus, a mutual antioxidant effect between diclofenac acid and these antioxidants prolongs the lifespan of VC, VC-12, VE and Trolox, respectively.

    Topics: Amidines; Ascorbic Acid; Chromans; Diclofenac; Dose-Response Relationship, Drug; Drug Combinations; Erythrocytes; Free Radical Scavengers; Free Radicals; Hemolysis; Humans; In Vitro Techniques; Solubility

2006
Icariin: a special antioxidant to protect linoleic acid against free-radical-induced peroxidation in micelles.
    The journal of physical chemistry. A, 2006, May-18, Volume: 110, Issue:19

    The objective in this work is to determine the antioxidant capacity and effectiveness of icariin (2-(4'-methoxylphenyl)-3-rhamnosido-5-hydroxyl-7-glucosido-8-(3'-methyl-2-butylenyl)-4-chromanone), the major component in herba epimedii being used widely in traditional Chinese medicine for the treatment of artherosclerosis and neuropathy, in which 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced peroxidation of linoleic acid (LH) in sodium dodecyl sulfate (SDS) acts as the experimental system. By containing an intramolecular hydrogen bond, icariin protects LH against AAPH-induced peroxidation of LH only in SDS, an anionic micelle. The number of trapping peroxyl radicals (LOO(*)), n, by icariin is just 0.0167 whereas alpha-tocopherol (TOH) and L-ascorbyl-6-laurate (VC-12) are 2.14 and 1.25, respectively, with reference to the n of 6-hydroxyl-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), 2.00. This is also related to how the intramolecular hydrogen bond enhances the bond dissociation enthalpy (BDE) of O-H in icariin. However, calculation of the inhibition rate constant, k(inh), a kinetic parameter to describe the reaction between the antioxidant and LOO(*), results in a k(inh) of icariin at about one magnitude larger than those of Trolox, TOH, and VC-12. This fact reveals that, by the view of kinetics, icariin is an antioxidant with much higher effectiveness. In addition, the antioxidant capacities of icariin used together with other antioxidants have been determined and the results indicate that the n of icariin decreases markedly while the n values of Trolox and TOH increase, even if the n of icariin is a negative value in the presence of VC-12. Furthermore, an analysis of k(inh) in this case reveals that the k(inh)(icariin) increases nearly one magnitude with the decrease of k(inh)(Trolox) and no remarkable change occurs for k(inh)(TOH). The negative value of k(inh)(icariin) in the presence of VC-12 can be regarded as the icariin functions as a prooxidant that can be rectified by VC-12 effectively. These findings implicate that the evaluation of antioxidant activity should not only focus on an n value, a thermodynamic possibility, but k(inh) and the charge property of the micelle should be also taken into account. To some extent, the latter factors are more important than the thermodynamic possibility.

    Topics: Algorithms; Antioxidants; Ascorbic Acid; Chromans; Flavonoids; Free Radicals; Hydrogen Bonding; Kinetics; Lauric Acids; Linoleic Acid; Lipid Peroxidation; Micelles; Sodium Dodecyl Sulfate

2006
Synthesis of 7-hydroxy-4-oxo-4H-chromene- and 7-hydroxychroman-2-carboxylic acid N-alkyl amides and their antioxidant activities.
    Archives of pharmacal research, 2006, Volume: 29, Issue:9

    A series of 7-hydroxy-4-oxo-4H-chromene- (3a - h) and 7-hydroxychroman-2-carboxylic acid N-alkyl amides (4a - g) were synthesized and their antioxidant activities were evaluated. While compounds 3a - h were less active, compounds 4a - g exhibited more potent inhibition of lipid peroxidation initiated by Fe2+ and ascorbic acid in rat brain homogenates. Among them, 7-hydroxychroman-2-carboxylic acid N-alkylamides (4e - g) bearing nonyl, decyl, and undecyl side chain exhibited 3 times more potent inhibition than trolox (1).

    Topics: Alkanes; Animals; Antioxidants; Ascorbic Acid; Biphenyl Compounds; Chromans; Free Radical Scavengers; Indicators and Reagents; Iron; Lipid Peroxidation; Picrates; Rats

2006
Oxidative stress influences cholesterol efflux in THP-1 macrophages: role of ATP-binding cassette A1 and nuclear factors.
    Cardiovascular research, 2006, 12-01, Volume: 72, Issue:3

    Understanding the mechanisms involved in oxidative stress-induced foam cell formation is of fundamental importance for atherosclerosis. Our aim was to characterize the effects of oxidative stress on key receptors of macrophage cholesterol homeostasis, on the nuclear transcription factors PPAR and LXR regulating their expression, and on macrophage cholesterol handling.. The incubation of macrophages derived from the human monocyte cell line THP-1 with iron (100 microm)/ascorbate (1000 microm) for a period of 4 h induced a strong peroxidation, as demonstrated by the elevation of malondialdehyde (220%, P < 0.001). The production of lipid peroxidation affected cholesterol efflux, which was probably due to decreased ABCAI gene and protein expression. On the other hand, cholesterol influx remained unchanged as did the mRNA and protein levels of SR-BI and CD36, important protein receptors that participate in cholesterol import. Experiments using RT-PCR showed that the ABCAI modulation was orchestrated by the nuclear receptors LXRalpha, LXRbeta, PPARalpha, and PPARgamma. Treatment with powerful antioxidants (Trolox and BHT) prevented the adverse effects of iron-ascorbate on cholesterol movement, conceivably supporting the role of oxidative stress.. Our results show that oxidative stress can directly be induced in macrophages and concomitantly impairs the expression of receptors involved in cholesterol flux, which could influence foam cell formation and atherosclerosis development.

    Topics: Antioxidants; Ascorbic Acid; Atherosclerosis; ATP Binding Cassette Transporter 1; ATP-Binding Cassette Transporters; Blotting, Western; Butylated Hydroxytoluene; CD36 Antigens; Cell Line; Cholesterol; Chromans; DNA-Binding Proteins; Foam Cells; Gene Expression; Homeostasis; Humans; Lipid Peroxidation; Liver X Receptors; Orphan Nuclear Receptors; Oxidative Stress; Peroxisome Proliferator-Activated Receptors; PPAR alpha; PPAR gamma; PPAR-beta; Receptors, Cytoplasmic and Nuclear; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Scavenger Receptors, Class B

2006
A pharmacological action of vitamin E unrelated to its antioxidant capacity.
    Methods and findings in experimental and clinical pharmacology, 2006, Volume: 28, Issue:8

    The pharmacological action of vitamin E on the mechanical activity of isolated guinea pig colonic smooth muscle was examined in normoxic and hypoxic conditions. In hypoxia, but not normoxia, alpha-tocopherol (1-160 microM) evoked rapid concentration-dependent contractions from the colon. This was also seen with other members of the vitamin E family, and potency measurements gave EC(50) values (microM) of 10.6 +/- 0.9 for D-alpha-tocopherol, 6.0 +/- 1.2 for D-beta-tocopherol, 7.5 +/- 0.7 for D-gamma-tocopherol, and 6.1 +/- 1.5 for D-delta-tocopherol. This order of potency for the components of the vitamin differs from previously studied bioassay systems and from their antioxidant activity. A range of potent natural and synthetic antioxidants was not active in this system. Compounds with structural similarities to the side chain of vitamin E produced similar stimulatory responses and some, like phytol, were more potent than the vitamin (EC(50): 1.0 +/- 0.2 microM), whereas ring structures related to the vitamin, like Trolox C, antagonized the stimulant responses in a concentration-dependent manner. Therefore, this model system measures, directly, vitamin E-induced responses through a mechanism that does not appear to be related to the known antioxidant capacity of these agents.

    Topics: alpha-Tocopherol; Animals; Antioxidants; Ascorbic Acid; Chromans; Colon, Transverse; Dose-Response Relationship, Drug; Female; Guinea Pigs; Hypoxia; In Vitro Techniques; Isometric Contraction; Male; Methacrylates; Muscle Contraction; Muscle, Smooth; Phytol; Pyrogallol; Quantitative Structure-Activity Relationship; Stereoisomerism; Temperature; Terpenes

2006
The combined effect of pycnogenol with ascorbic acid and trolox on the oxidation of lipids and proteins.
    General physiology and biophysics, 2006, Volume: 25, Issue:4

    Pycnogenol (PYC), a procyanidin-rich extract of French maritime pine bark (Pinus pinaster) has strong antioxidant potential and promotes cellular health. The aim of this study was to investigate a possible cooperation of natural antioxidant PYC with synthetic antioxidants ascorbic acid and trolox in the model system of lipid peroxidation determined as conjugated dienes formation in liposomes and on the oxidation of proteins (in BSA and plasma proteins) determined as protein carbonyls. The present study shows that PYC and trolox significantly increased inhibition of lipid peroxidation initiated by copper acetate and tert-butylhydroperoxide in concentration and time dependence compared with untreated unilamellar liposomes. PYC and trolox added simultaneously to the oxidized liposomes exerted an additive preventive effect. PYC s inhibitory effect on formation of carbonyl compounds in BSA and plasma proteins, oxidized by two oxidative systems--H2O2/FeSO4 and HOCl, were studied in co-operation with other synthetic antioxidants--ascorbic acid and trolox. We found the synergistic or additive effect of PYC with mentioned antioxidants.

    Topics: Ascorbic Acid; Chromans; Drug Combinations; Flavonoids; Lipids; Oxidation-Reduction; Plant Extracts; Proteins

2006
Antioxidant-mediated augmentation of ozone-induced membrane oxidation.
    Free radical biology & medicine, 2005, Feb-15, Volume: 38, Issue:4

    The pulmonary epithelial lining fluid (ELF) contains substrates, e.g., ascorbic acid (AH2), uric acid (UA), glutathione (GSH), proteins, and unsaturated lipids, which undergo facile reaction with inhaled ozone (O3). Reactions near the ELF gas/liquid interface likely provide the driving force for O3 absorption ("reactive absorption") and constrain O3 diffusion to the underlying epithelium. To investigate the potential mechanisms wherein O3/ELF interactions may induce cellular damage, we utilized a red cell membrane (RCM) model intermittently covered by an aqueous film to mimic the lung surface compartmentation, and evaluated exposure-mediated loss of acetylcholinesterase activity (AChE) and TBARS accumulation. In the absence of aqueous reactants, O3 exposure induced no detectable changes in AChE or TBARS. AH2 and GSH preferentially induced oxidative damage in a dose-dependent fashion. AH2-mediated RCM oxidation was not inhibited by superoxide dismutase, catalase, mannitol, or Fe chelators. O3 reaction with UA, Trolox, or albumin produced no RCM oxidation but oxidation occurred when AH2 was combined with UA or albumin. Rat bronchoalveolar lavage fluid (BALF) also induced RCM oxidation. However, in vivo O3 exposure dampened the extent of BALF-mediated RCM oxidation. Although we cannot completely rule out O3 diffusion to the RCM, product(s) derived from O3 + AH2/GSH reactions (possibly O3*- or 1O2) likely initiated RCM oxidation and may suggest that in vivo, such secondary species account for O3 permeation through the ELF leading to cellular perturbations.

    Topics: Acetylcholinesterase; Aldehydes; Animals; Antioxidants; Ascorbic Acid; Chromans; Erythrocyte Membrane; Glutathione; Humans; L-Lactate Dehydrogenase; Male; Oxidation-Reduction; Ozone; Rats; Rats, Sprague-Dawley; Uric Acid

2005
Chemical composition, antimicrobial and antioxidant activity of the essential oil of Teucrium marum (Lamiaceae).
    Journal of ethnopharmacology, 2005, Apr-08, Volume: 98, Issue:1-2

    The chemical composition of the essential oil obtained from Teucrium marum subsp. marum (Lamiaceae) was analysed by GC/MS and 30 components were identified. Isocaryophyllene (20.24%), beta-bisabolene (14.73%), beta-sesquiphellandrene (11.27%), alpha-santalene (10.97%), dolichodial (9.38%) and, alpha-caryophyllene (7.18%) were the main components. The antimicrobial activity of the essential oil was assayed against four phytopathogenic fungi and Rhyzoctonia solani resulted to be the most sensitive microorganism with a MIC value of 250 ppm. The antioxidant activity of the essential oil was evaluated using the DPPH test, 5-lipoxygenase test and luminol/xanthine/xanthine oxidase chemiluminescence assay.

    Topics: Antioxidants; Ascorbic Acid; Biphenyl Compounds; Butylated Hydroxytoluene; Chromans; Drug Evaluation, Preclinical; Fatty Acids; Free Radical Scavengers; Fungicides, Industrial; Inhibitory Concentration 50; Microbial Sensitivity Tests; Mitosporic Fungi; Monoterpenes; Mycoses; Picrates; Plant Components, Aerial; Plant Diseases; Plant Oils; Sesquiterpenes; Superoxides; Teucrium

2005
Hypoxic augmentation of Ca2+ channel currents requires a functional electron transport chain.
    The Journal of biological chemistry, 2005, Jun-10, Volume: 280, Issue:23

    The incidence of Alzheimer disease is increased following ischemic episodes, and we previously demonstrated that following chronic hypoxia (CH), amyloid beta (Abeta) peptide-mediated increases in voltage-gated L-type Ca(2+) channel activity contribute to the Ca(2+) dyshomeostasis seen in Alzheimer disease. Because in certain cell types mitochondria are responsible for detecting altered O(2) levels we examined the role of mitochondrial oxidant production in the regulation of recombinant Ca(2+) channel alpha(1C) subunits during CH and exposure to Abeta-(1-40). In wild-type (rho(+)) HEK 293 cells expressing recombinant L-type alpha(1C) subunits, Ca(2+) currents were enhanced by prolonged (24 h) exposure to either CH (6% O(2)) or Abeta-(1-40) (50 nm). By contrast the response to CH was absent in rho(0) cells in which the mitochondrial electron transport chain (ETC) was depleted following long term treatment with ethidium bromide or in rho(+) cells cultured in the presence of 1 microm rotenone. CH was mimicked in rho(0) cells by the exogenous production of O2(-.). by xanthine/xanthine oxidase. Furthermore Abeta-(1-40) enhanced currents in rho(0) cells to a degree similar to that seen in cells with an intact ETC. The antioxidants ascorbate (200 microm) and Trolox (500 microm) ablated the effect of CH in rho(+) cells but were without effect on Abeta-(1-40)-mediated augmentation of Ca(2+) current in rho(0) cells. Thus oxidant production in the mitochondrial ETC is a critical factor, acting upstream of amyloid beta peptide production in the up-regulation of Ca(2+) channels in response to CH.

    Topics: Alzheimer Disease; Amyloid beta-Peptides; Antioxidants; Ascorbic Acid; Biological Transport; Brain Ischemia; Calcium; Calcium Channels; Cell Line; Chromans; Electron Transport; Electrons; Electrophysiology; Ethidium; Humans; Hypoxia; Immunohistochemistry; Mitochondria; Neurodegenerative Diseases; Oxidants; Oxygen; Peptides; Reactive Oxygen Species; Rotenone; Superoxides; Transfection; Up-Regulation; Xanthine Oxidase

2005
Neuroprotectant effects of iso-osmolar D-mannitol to prevent Pacific ciguatoxin-1 induced alterations in neuronal excitability: a comparison with other osmotic agents and free radical scavengers.
    Neuropharmacology, 2005, Volume: 49, Issue:5

    The basis for the neuroprotectant effect of D-mannitol in reducing the sensory neurological disturbances seen in ciguatera poisoning, is unclear. Pacific ciguatoxin-1 (P-CTX-1), at a concentration 10 nM, caused a statistically significant swelling of rat sensory dorsal root ganglia (DRG) neurons that was reversed by hyperosmolar 50 mM D-mannitol. However, using electron paramagnetic resonance (EPR) spectroscopy, it was found that P-CTX-1 failed to generate hydroxyl free radicals at concentrations of toxin that caused profound effects on neuronal excitability. Whole-cell patch-clamp recordings from DRG neurons revealed that both hyper- and iso-osmolar 50 mM D-mannitol prevented the membrane depolarisation and repetitive firing of action potentials induced by P-CTX-1. In addition, both hyper- and iso-osmolar 50 mM D-mannitol prevented the hyperpolarising shift in steady-state inactivation and the rise in leakage current through tetrodotoxin (TTX)-sensitive Na(v) channels, as well as the increased rate of recovery from inactivation of TTX-resistant Na(v) channels induced by P-CTX-1. D-Mannitol also reduced, but did not prevent, the inhibition of peak TTX-sensitive and TTX-resistant I(Na) amplitude by P-CTX-1. Additional experiments using hyper- and iso-osmolar D-sorbitol, hyperosmolar sucrose and the free radical scavenging agents Trolox and L-ascorbic acid showed that these agents, unlike D-mannitol, failed to prevent the effects of P-CTX-1 on spike electrogenesis and Na(v) channel gating. These selective actions of D-mannitol indicate that it does not act purely as an osmotic agent to reduce swelling of nerves, but involves a more complex action dependent on the Na(v) channel subtype, possibly to alter or reduce toxin association.

    Topics: Algorithms; Ascorbic Acid; Cell Size; Chromans; Ciguatoxins; Diuretics; Electron Spin Resonance Spectroscopy; Electrophysiology; Free Radical Scavengers; Ganglia, Spinal; Ion Channel Gating; Mannitol; Membrane Potentials; Neurons, Afferent; Neuroprotective Agents; Osmolar Concentration; Patch-Clamp Techniques; Sodium Channel Blockers; Sodium Channels; Sorbitol; Tetrodotoxin

2005
Antioxidants prevented oxidative injury of SR induced by Fe2+/H2O2/ascorbate system but failed to prevent Ca2+-ATPase activity decrease.
    BioFactors (Oxford, England), 2005, Volume: 24, Issue:1-4

    Dysfunction of sarcoplasmic reticulum (SR) Ca2+-ATPase induced by oxidative stress may be a contributing factor to the development of serious age related diseases. Incubation of sarcoplasmic reticulum (SR) vesicles of rabbit skeletal muscles with Fe2+/H2O2/ascorbate decreased the SH group content of SR approximately to 35% and Ca2+-ATPase activity to 50% of control not oxidized sample. Protein carbonyls increased twofold, lipid peroxidation was also significantly elevated. The antioxidant effects of trolox, the pyridoindole derivative stobadine and of the standardized extracts from bark of Pinus Pinaster PycnogenolR (Pyc) and from leaves of Ginkgo biloba (EGb 761) were studied on oxidatively injured SR. All antioxidants exerted preventive effects against the oxidized lipids and protein SH groups of SR vesicles. Trolox and stobadine did not influence protein carbonyl formation, while flavonoid extracts prevented carbonyl generation, probably by binding to protein. The preventive effects of the antioxidants studied on lipids and protein SH groups were however not associated with protection of Ca2+-ATPase activity. Stobadine and trolox exerted no effect on enzyme activity, Pyc and EGb 761 enhanced the inhibitory effect of Ca2+-ATPase activity in oxidatively injured SR. Concluding, under the conditions of oxidative stress induced by Fe2+/H2O2/ascorbate against SR of rabbit skeletal muscle, the agents studied demonstrated antioxidant effects yet failed to protect Ca2+-ATPase activity.

    Topics: Animals; Antioxidants; Ascorbic Acid; Calcium-Transporting ATPases; Carbolines; Chromans; Ferrous Compounds; Flavonoids; Ginkgo biloba; Hydrogen Peroxide; Muscle, Skeletal; Oxidative Stress; Pinus; Plant Bark; Plant Leaves; Rabbits; Sarcoplasmic Reticulum; Sulfhydryl Compounds

2005
4-Hydroxy tempol-induced impairment of mitochondrial function and augmentation of glucose transport in vascular endothelial and smooth muscle cells.
    Biochemical pharmacology, 2004, May-15, Volume: 67, Issue:10

    The water-soluble and cell permeable nitroxide derivative 4-hydroxy tempol (TPL) has been shown to reduce or ameliorate oxidative stress-induced dysfunction and damage in vascular endothelial cells. We studied the effects of TPL on glucose transport and metabolism in bovine aortic endothelial (VEC) and smooth muscle cells (VSMC) under normal and high glucose conditions. Normally, these cells operate an autoregulatory protective mechanism that limits the rate of glucose transport under hyperglycemic conditions by decreasing the cell content of their typical glucose transporter GLUT-1 mRNA and protein as well as its plasma membrane abundance. TPL augmented the rate of glucose transport both under normo- and hyperglycemic conditions by increasing GLUT-1 mRNA and protein content and its plasma membrane abundance in both types of cells, leading to an increased flux of glucose into the cells. These effects were found related to ROS-generating and oxidant activities of TPL and to a decreased rate of mitochondrial ATP production under both normo- and hyperglycemic conditions. Since impaired mitochondrial functions, and in particular decreased rate of ATP production, augment the expression of GLUT-1 protein and glucose transport and metabolism, we suggest that the stimulatory effects of TPL in vascular cells results from its unfavorable interactions in the mitochondrion. It is therefore suggested that effects of TPL in cells of cardiovascular system be evaluated in parallel to its adverse effects on glucose and energy metabolism.

    Topics: Acetylcysteine; Adenosine Triphosphate; Animals; Antioxidants; Ascorbic Acid; Biological Transport; Cattle; Cell Membrane; Chromans; Cyclic N-Oxides; Endothelium, Vascular; Glucose; Glucose Transporter Type 1; Guanidines; Mitochondria; Monosaccharide Transport Proteins; Myocytes, Smooth Muscle; RNA, Messenger; Spin Labels

2004
Hydroxyl radical generation caused by the reaction of singlet oxygen with a spin trap, DMPO, increases significantly in the presence of biological reductants.
    Free radical research, 2004, Volume: 38, Issue:4

    Photosensitizers newly developed for photodynamic therapy of cancer need to be assessed using accurate methods of measuring reactive oxygen species (ROS). Little is known about the characteristics of the reaction of singlet oxygen (1O2) with spin traps, although this knowledge is necessary in electron spin resonance (ESR)/spin trapping. In the present study, we examined the effect of various reductants usually present in biological samples on the reaction of 1O2 with 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). The ESR signal of the hydroxyl radical (*OH) adduct of DMPO (DMPO-OH) resulting from 1O2-dependent generation of *OH strengthened remarkably in the presence of reduced glutathione (GSH), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), ascorbic acid, NADPH, etc. A similar increase was observed in the photosensitization of uroporphyrin (UP), rose bengal (RB) or methylene blue (MB). Use of 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide (DEPMPO) as a spin trap significantly lessened the production of its *OH adduct (DEPMPO-OH) in the presence of the reductants. The addition of DMPO to the DEPMPO-spin trapping system remarkably increased the signal intensity of DEPMPO-OH. DMPO-mediated generation of *OH was also confirmed utilizing the hydroxylation of salicylic acid (SA). These results suggest that biological reductants enhance the ESR signal of DMPO-OH produced by DMPO-mediated generation of *OH from 1O2, and that spin trap-mediated *OH generation hardly occurs with DEPMPO.

    Topics: Antioxidants; Ascorbic Acid; Azides; Chromans; Cyclic N-Oxides; Electron Spin Resonance Spectroscopy; Ethanol; Glutathione; Hydroxyl Radical; Manganese; Oxygen; Reactive Oxygen Species; Salicylic Acid; Spin Trapping; Superoxides

2004
Inherent redox properties of diesel exhaust particles: catalysis of the generation of reactive oxygen species by biological reductants.
    Toxicological sciences : an official journal of the Society of Toxicology, 2004, Volume: 81, Issue:1

    The toxicity of diesel exhaust particles (DEP) can be due to the particle itself, extractable components, or both. Many studies focus on the biological properties of DEP-extractable components although it is possible that chemical properties inherent to the DEP itself can lead to toxicity. Thus, an examination of the chemistry inherent to DEP was carried out. Herein, we report that DEP are capable of catalyzing the consumption of O2 (monitored using a Clarke electrode) by ascorbate and thiols leading to the generation of reactive oxygen species. Consistent with the idea that DEP are capable of catalyzing the generation of reactive oxygen species, they were also found to catalyze DNA strand breakage via an O2- and reductant-dependent process. Significantly, extraction of DEP with either organic solvent (methylene chloride) or acid (aqueous HCl) did little to abrogate this chemistry. Finally, using electron paramagnetic spectrometry (EPR), DEP were found to have paramagnetic properties. The paramagnetic character of DEP may be important to their ability to catalyze the formation of reactive oxygen species and at least partially responsible for their toxicity. These findings indicate that studies that primarily consider or examine particle extracts as the toxic components of DEP may be insufficient in describing the toxicity associated with DEP exposure.

    Topics: Algorithms; Antioxidants; Ascorbic Acid; Catalysis; Chromans; DNA; DNA Damage; Electron Spin Resonance Spectroscopy; Glutathione; In Situ Nick-End Labeling; NAD; Oxidation-Reduction; Oxygen Consumption; Plasmids; Reactive Oxygen Species; Solvents; Vehicle Emissions

2004
The increase in human plasma antioxidant capacity after apple consumption is due to the metabolic effect of fructose on urate, not apple-derived antioxidant flavonoids.
    Free radical biology & medicine, 2004, Jul-15, Volume: 37, Issue:2

    Regular fruit consumption lowers the risk of cardiovascular diseases and certain cancers, which has been attributed in part to fruit-derived antioxidant flavonoids. However, flavonoids are poorly absorbed by humans, and the increase in plasma antioxidant capacity observed after consumption of flavonoid-rich foods often greatly exceeds the increase in plasma flavonoids. In the present study, six healthy subjects consumed five Red Delicious apples (1037 +/- 38 g), plain bagels (263.1 +/- 0.9 g) and water matching the carbohydrate content and mass of the apples, and fructose (63.9 +/- 2.9 g) in water matching the fructose content and mass of the apples. The antioxidant capacity of plasma was measured before and up to 6 h after food consumption as ferric reducing antioxidant potential (FRAP), without or with ascorbate oxidase treatment (FRAPAO) to estimate the contribution of ascorbate. Baseline plasma FRAP and FRAPAO were 445 +/- 35 and 363 +/- 35 microM trolox equivalents, respectively. Apple consumption caused an acute, transient increase in both plasma FRAP and FRAPAO, with increases after 1 h of 54.6 +/- 8.7 and 61.3 = 17.2 microM trolox equivalents, respectively. This increase in plasma antioxidant capacity was paralleled by a large increase in plasma urate, a metabolic antioxidant, from 271 +/- 39 microM at baseline to 367 +/- 43 microM after 1 h. In contrast, FRAP and FRAPAO time-dependently decreased after bagel consumption, together with urate. Consumption of fructose mimicked the effects of apples with respect to increased FRAP, FRAPAO, and urate, but not ascorbate. Taken together, our data show that the increase in plasma antioxidant capacity in humans after apple consumption is due mainly to the well-known metabolic effect of fructose on urate, not apple-derived antioxidant flavonoids.

    Topics: Antioxidants; Ascorbic Acid; Bread; Carbohydrates; Chromans; Flavonoids; Free Radicals; Fructose; Fruit; Humans; Malus; Nutritional Physiological Phenomena; Time Factors; Uric Acid; Vitamins; Water

2004
Detection of lead-induced oxidative stress in the rat epididymis by chemiluminescence.
    Chemosphere, 2004, Volume: 57, Issue:10

    It has been shown that lead (Pb) is able to induce lipid peroxidation, one of the main manifestations of oxidative stress. In this study we examined the relationship between chronic Pb exposure and level of reactive oxygen species (ROS) in reproductive system tissues of sexually mature male Wistar rats. One group of animals (control, K) was allowed to drink distilled water, the second group (Pb) was allowed to drink freely 1% aqueous solution of lead acetate. Another groups had a following supplements: rats were allowed to drink distilled water containing vitamin C (vit C) at concentration of 500 mg/l or Trolox (a vitamin E analog) at concentration of 48 mg/l or vit C (500 mg/l) + Trolox (48 mg/l). The similar groups among Pb-treated animals were examined after treatment with the same vitamins and using the same vitamin doses, dissolved in 1% aqueous solution of lead acetate. In all cases the time of drinking was 6 months. It was found that lead content in samples of tissues from testis, epididymis and in a whole blood in Pb- and Pb with antioxidants treated rats was significantly elevated. Chemiluminescence (CL) emitted by the Pb-treated tissues was significantly higher when compared to the light emission by tissues isolated from the animals of control group. The increase in the CL caused by lead occurs in the following increasing order within the studied tissues: cauda of epididymis < testis < caput of epididymis (19%, 39% and 51%, respectively). Dietary vit C supplementation to the Pb-treated rats for 6 months period decreased the CL from caput of epididymis, cauda of epididymis and testis (by 43%, 24%, 39%, respectively) more effectively in comparison to the control group (35%, 17%, 33%, respectively). Also stronger quenching effect on the light emission from the above mentioned tissues after Trolox supplementation was observed in the Pb-treated group (42%, 21%, 35%, respectively) than in the control group (23%, 13%, 13% respectively). The combination of both antioxidants treatments (vit C and Trolox) did not give a higher significant quenching effect compared to the treatment with the vitamins separately. No ultrastructural changes were found in the seminiferous epithelium of Pb-treated animals. However, we found abnormalities in ultrastructure of epididymal epithelial cells and epididymal spermatozoa in rats of Pb-treated groups. These findings provide ex vivo evidence that Pb causes oxidative cellular damage in reproductive system tissues of adult

    Topics: Analysis of Variance; Animals; Antioxidants; Ascorbic Acid; Body Weight; Chromans; Epididymis; Lead; Luminescent Measurements; Male; Organ Size; Oxidative Stress; Rats; Rats, Wistar; Reactive Oxygen Species; Testis

2004
In vitro effects of ascorbate and Trolox on the biocompatibility of dental restorative materials.
    Biomaterials, 2003, Volume: 24, Issue:1

    Previous in vitro studies on the cytotoxicity of eight dental restorative materials including composites, compomers, resin-modified glass ionomer cements and glass ionomer cements have demonstrated a depletion of intracellular glutathione in gingival fibroblasts incubated with eluates of these materials and a protective effect of N-acetylcysteine. In the present study, we investigate the effects of two other antioxidants: ascorbate and Trolox. It was found that Trolox reduced the cytotoxicity induced by resin-based biomaterial eluates. In contrast, ascorbate increased in a dose-dependent manner the toxic effect of all eluates except for Z100 MP and Tetric flow (composites). The effect of D-mannitol was studied for GC FUJI II and was found to neutralize the additional toxic effect of ascorbate. Ascorbate increased the depletion of intracellular glutathione of these dental material eluates (between 17% and 24%, depending on the material). Quantification of metal ions in the dental material eluates showed the presence of significant amounts of aluminum and iron in GC FUJI II > photac fil > GC FUJI II LC > F2000. The mechanism of this increased cytotoxicity could be explained by the Fenton reaction resulting from the pro-oxidant effect of ascorbate in the presence of iron (transition metal ions) and/or aluminum.

    Topics: Aluminum; Ascorbic Acid; Cell Survival; Cells, Cultured; Chromans; Dental Materials; Dental Restoration, Permanent; Fibroblasts; Free Radical Scavengers; Gingiva; Glutathione; Humans; In Vitro Techniques; Iron; Mannitol; Materials Testing; Metals

2003
Protection against oxidative protein damage induced by metal-catalyzed reaction or alkylperoxyl radicals: comparative effects of melatonin and other antioxidants.
    Biochimica et biophysica acta, 2003, Mar-17, Volume: 1620, Issue:1-3

    Melatonin is a well-known hydroxyl radical (*OH) scavenger that protects DNA and lipids from free radical attack. In this paper, we studied the ability of melatonin to prevent oxidative damage to bovine serum albumin (BSA) induced by two different paradigms: the metal-catalyzed oxidation (MCO) induced by Cu(2+)/H(2)O(2) and the alkoxyl and alkylperoxyl radicals formed by the azo initiator 2,2'-azobis(2-amidinopropane) hydrochloride (AAPH, 40 mM). The protective effects of melatonin were compared with 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (trolox), glutathione (GSH), ascorbate, 3,4',5-trihydroxy-trans-stilbene (resveratrol, 0.1 microM-4 mM) and mannitol (50 microM-100 mM). Melatonin efficiently prevented protein modification induced by both models, as assayed by polyacrylamide gel electrophoresis and carbonyl content. Both trolox and ascorbate had an obvious pro-oxidant effect in the Cu(2+)/H(2)O(2) model, whereas both prevented BSA damage induced by AAPH. In the MCO model, the efficacy of GSH in terms of protein protection was higher than melatonin at relatively high concentrations (250 microM-4 mM); however, at lower concentrations (50-250 microM), the efficacy of melatonin was superior to GSH. D-Mannitol (50 microM-100 mM) and resveratrol did not protect BSA from the site-specific damage induced by Cu(2+)/H(2)O(2). On the other hand, the relative protective efficiency in the AAPH model was melatonin approximately trolox>GSH>ascorbate.

    Topics: Amidines; Animals; Antioxidants; Ascorbic Acid; Cattle; Chromans; Copper; Electrophoresis, Polyacrylamide Gel; Hydrogen Peroxide; Melatonin; Metals; Oxidants; Peroxides; Serum Albumin, Bovine; Time Factors

2003
Antioxidant activity of melatonin in Chinese hamster ovarian cells: changes in cellular proliferation and differentiation.
    Biochemical and biophysical research communications, 2003, Mar-14, Volume: 302, Issue:3

    Melatonin is an endogenously generated molecule with free radical scavenging and antioxidant properties. Here, we studied the antiproliferative role of melatonin and other antioxidants on transformed Chinese hamster ovarian cells. Melatonin reduces cell proliferation in a dose- and time-dependent manner. Natural antioxidants which appear in edible plants including resveratrol and vitamin E mimicked the effect of melatonin. Flow cytometer analysis revealed that melatonin treatment reduces the number of cells in S-phase and increases cells in both G0/G1 and G2/M gaps. In addition, melatonin, as well as trolox, caused a clear morphological change by inducing the cells to become spindle shaped and fibroblast-like. Its effect is a reversible phenomenon that disappeared when melatonin was withdrawn from the culture medium. GSH levels are increased after melatonin treatment but pharmacologically blockade of GSH synthesis did not abolish melatonin's antiproliferative effect. Reduction of cell proliferation and the apparent induction of cell differentiation overlapped with melatonin's ability to change the intracellular redox state of CHO cells. We conclude that the cellular redox state may be involved in cellular transformation caused by antioxidants such as melatonin and trolox.

    Topics: Animals; Antioxidants; Ascorbic Acid; Cell Differentiation; Cell Division; Cells, Cultured; CHO Cells; Chromans; Coloring Agents; Cricetinae; Dose-Response Relationship, Drug; Flow Cytometry; G1 Phase; G2 Phase; Glutathione; Glutathione Disulfide; Mitosis; Oxidation-Reduction; Reactive Oxygen Species; Resting Phase, Cell Cycle; Resveratrol; S Phase; Stilbenes; Tetrazolium Salts; Thiazoles; Time Factors; Trypan Blue

2003
Mechanistic and comparative studies of melatonin and classic antioxidants in terms of their interactions with the ABTS cation radical.
    Journal of pineal research, 2003, Volume: 34, Issue:4

    Melatonin and classic antioxidants possess the capacity to scavenge ABTSb+ with IC50s of 4, 11, 15.5, 15.5, 17 and 21 microm for melatonin, glutathione, vitamin C, trolox, NADH and NADPH, respectively. In terms of scavenging ABTSb+, melatonin exhibits a different profile than that of the classic antioxidants. Classic antioxidants scavenge one or less ABTSb+, while each melatonin molecule can scavenge more than one ABTSb+, probably with a maximum of four. Classic antioxidants do not synergize when combined in terms of scavenging ABTSb+. However, a synergistic action is observed when melatonin is combined with any of the classic antioxidants. Cyclic voltammetry indicates that melatonin donates an electron at the potential of 715 mV. The scavenging mechanism of melatonin on ABTSb+ may involve multiple-electron donations via intermediates through a stepwise process. Intermediates including the melatoninyl cation radical, the melatoninyl neutral radical and cyclic 3-hydroxymelatonin (cyclic 3-OHM) and N1-acetyl-N2-formyl-5-methoxykynuramine (AFMK) seem to participate in these reactions. More interestingly, the pH of the solution dramatically modifies the ABTSb+ scavenging capacity of melatonin while pH changes have no measurable influence on the scavenging activity of classic antioxidants. An acidic pH markedly reduces the ABTSb+ scavenging capacity of melatonin while an increased pH promotes the interaction of melatonin and ABTSb+. The major melatonin metabolites that develop when melatonin interacts with ABTSb+ are cyclic 3-OHM and AFMK. Cyclic 3-OHM is the intermediate between melatonin and AFMK, and cyclic 3-OHM also has the ability to scavenge ABTSb+. Melatonin and the metabolites which are generated via the interaction of melatonin with ABTSb+, i.e. the melatoninyl cation radical, melatoninyl neutral radical and cyclic 3-OHM, all scavenge ABTSb+. This unique cascade action of melatonin, in terms of scavenging, increases its efficiency to neutralized ABTSb+; this contrasts with the effects of the classic antioxidants.

    Topics: Antioxidants; Ascorbic Acid; Benzothiazoles; Cations; Chromans; Chromatography, High Pressure Liquid; Electrochemistry; Free Radical Scavengers; Free Radicals; Glutathione; Hydrogen-Ion Concentration; Kynuramine; Mechanics; Melatonin; NAD; NADP; Sulfonic Acids

2003
Antioxidants attenuate myocyte apoptosis and improve cardiac function in CHF: association with changes in MAPK pathways.
    American journal of physiology. Heart and circulatory physiology, 2003, Volume: 285, Issue:2

    Antioxidant vitamins reduce cardiac oxidative stress and cardiomyocyte apoptosis produced by exogenous norepinephrine (NE) and attenuate cardiac dysfunction in animals with pacing-induced congestive heart failure (CHF). This study was carried out to determine whether the mitogen-activated protein kinase (MAPK) signal transduction pathways are involved in oxidative stress-induced myocyte apoptosis. Rabbits with rapid pacing-induced CHF and sham operation were randomized to receive either a combination of antioxidant vitamins (beta-carotene, ascorbic acid, and alpha-tocopherol), alpha-tocopherol alone, or placebo for 8 wk. Compared with sham-operated animals, CHF animals exhibited increased oxidative stress as evidenced by decreased myocardial reduced-to-oxidized glutathione (GSH/GSSG) ratio (27 +/- 7 vs. 143 +/- 24, P < 0.05), myocyte apoptosis (77 +/- 18 vs. 17 +/- 4 apoptotic nuclei/10,000 cardiomyocytes, P < 0.05), increased total and phosphorylated c-Jun NH2-terminal protein kinase (p-JNK; 1.95 +/- 0.14 vs. 1.04 +/- 0.04 arbitrary units, P < 0.05) and phosphorylated p38 kinase (p-p38), and decreased phosphorylated extracellular signal-regulated kinase (p-ERK). Administration of antioxidant vitamins and alpha-tocopherol attenuated oxidative stress, myocyte apoptosis, and cardiac dysfunction, with reversal of the changes of total JNK, p-JNK, and p-ERK in CHF. Furthermore, because NE infusion produced changes of JNK, p-p38, and p-ERK similar to those in CHF, we conclude that NE may play an important role in the production of oxidative stress, MAPK activation, and myocyte apoptosis in CHF.

    Topics: alpha-Tocopherol; Animals; Antibodies, Monoclonal; Antioxidants; Apoptosis; Ascorbic Acid; beta Carotene; Blotting, Western; Cardiac Pacing, Artificial; Chromans; DNA, Single-Stranded; Glutathione; Heart Failure; MAP Kinase Signaling System; Mitogen-Activated Protein Kinase 8; Mitogen-Activated Protein Kinase 9; Mitogen-Activated Protein Kinases; Myocardium; Myocytes, Cardiac; Norepinephrine; Oxidative Stress; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Rabbits; Superoxide Dismutase; Sympathomimetics

2003
Photoprotective properties of a hydrophilic extract of the fern Polypodium leucotomos on human skin cells.
    Journal of photochemistry and photobiology. B, Biology, 2003, Volume: 70, Issue:1

    The effect of a hydrophilic extract of the fern Polypodium leucotomos (PLE) has been investigated in terms of photoprotection against UV-induced cell damage. PLE efficiently preserved human fibroblast survival and restored their proliferative capability when the cells were exposed to UVA light. This effect was specific and dose-dependent. Photoprotection was not restricted to fibroblasts, as demonstrated by its effect on survival and proliferation of the human keratinocyte cell line HaCat. Finally, treatment of the cells with PLE prevented UV-induced morphological changes in human fibroblasts, namely disorganisation of F-actin-based cytoskeletal structures, coalescence of the tubulin cytoskeleton and mislocalization of adhesion molecules such as cadherins and integrins. Our in vitro results demonstrate the photoprotective effect of PLE on human cells and support its use in the preventive treatment of sunburning and skin pathologies associated with UV-mediated damage.

    Topics: Acetylcysteine; Ascorbic Acid; Cell Adhesion; Cell Division; Cell Line; Cell Survival; Cells, Cultured; Chromans; Cytoskeleton; Fibroblasts; Humans; Plant Extracts; Polypodium; Protective Agents; Skin; Ultraviolet Rays

2003
Activity and location of olive oil phenolic antioxidants in liposomes.
    Chemistry and physics of lipids, 2003, Volume: 124, Issue:1

    The antioxidant activity of hydroxytyrosol, hydroxytyrosol acetate, oleuropein, 3,4-dihydroxyphenylelenolic acid (3,4-DHPEA-EA) and 3,4-dihydroxyphenylelenolic acid dialdehyde (3,4-DHPEA-EDA) towards oxidation initiated by 2,2'-azobis(2-amidinopropane) hydrochloride in a soybean phospholipid liposome system was studied. The antioxidant activity of these olive oil phenols was similar and the duration of the lag phase was almost twice that of alpha-tocopherol. Trolox, a water-soluble analogue of alpha-tocopherol, showed the worst antioxidant activity. However, oxidation before the end of the lag phase was inhibited less effectively by the olive oil phenols than by alpha-tocopherol and Trolox. Synergistic effects (11-20% increase in lag phase) were observed in the antioxidant activity of combinations of alpha-tocopherol with olive oil phenols both with and without ascorbic acid. Fluorescence anisotropy of probes and fluorescence quenching studies showed that the olive oil phenols did not penetrate into the membrane, but their effectiveness as antioxidants showed they were associated with the surface of the phospholipid bilayer.

    Topics: alpha-Tocopherol; Amidines; Antioxidants; Ascorbic Acid; Benzothiazoles; Chromans; Fluorescence Polarization; Glycine max; Kinetics; Liposomes; Membrane Fluidity; Olive Oil; Oxidation-Reduction; Phenols; Plant Oils; Sulfonic Acids

2003
Oxidative damage to catalase induced by peroxyl radicals: functional protection by melatonin and other antioxidants.
    Free radical research, 2003, Volume: 37, Issue:5

    Thermal decomposition by the azo initiator 2,2' azobis(2-amidinopropane) dihydrochloride (AAPH) has been widely used as a water-soluble source of free radical initiators capable of inducing lipid peroxidation and protein damage. Here, in a lipid-free system, AAPH alone (40 mM) rapidly induced protein modification and inactivation of the enzyme catalase (EC 1.11.1.6). Using SDS-PAGE, it was shown that protein band intensity is dramatically reduced after 4 h of incubation with AAPH, leading to protein aggregation. Several antioxidants including melatonin, glutathione (GSH) and trolox prevented catalase modification when used at a 250 microM concentration whereas ascorbate was only effective at 1 mM concentration. All the antioxidants tested reduced carbonyl formation although melatonin was the most effective in this regard. Enzyme inactivation caused by AAPH was also significantly reduced by the antioxidants and again melatonin was more efficient than the other antioxidants used in this study. Results shown here demonstrate that alkyl peroxyl radicals inactivate catalase and reduce the effectiveness of cells to defend against free radical damage; the damage to catalase can be prevented by antioxidants, especially melatonin.

    Topics: Amidines; Antioxidants; Ascorbic Acid; Catalase; Chromans; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Glutathione; Melatonin; Oxidants; Oxidation-Reduction; Peroxides; Time Factors

2003
Cadmium induces apoptosis in anterior pituitary cells that can be reversed by treatment with antioxidants.
    Toxicology and applied pharmacology, 2003, Jul-01, Volume: 190, Issue:1

    Cadmium (Cd(2+)) is an ubiquitous toxic metal that is involved in a variety of pathological conditions. Several reports indicate that Cd(2+) alters normal pituitary hormone secretion; however, little is known about the mechanisms that induce this misregulation. This paper reports the effect of Cd(2+) on anterior pituitary cell viability and its relation to prolactin secretion. Cd(2+) concentrations above 10 microM were found to be cytotoxic for pituitary cells. Morphological studies as well as DNA ladder fragmentation and caspase activation showed that Cd(2+)-treated cells undergo apoptosis. Even though several hours were needed to detect Cd(2+)-induced cytotoxicity, the effect of the metal became irreversible very quickly, requiring only 3 h of treatment. Prolactin release (measured at 48 h) was inhibited when the cells were exposed to Cd(2+) for 1 h, before any change in cell viability was observed. The antioxidants N-acetyl-cysteine and Trolox (a hydrosoluble derivative of vitamin E), but not ascorbic acid, reversed both Cd(2+)-mediated cytotoxicity and the inhibition of prolactin release, supporting the involvement of oxidative stress in the mechanism of Cd(2+) action. In summary, the present work demonstrates that Cd(2+) is cytotoxic for anterior pituitary cells, that this effect is due to an induction of apoptosis, and that it can be reversed by antioxidants.

    Topics: Acetylcysteine; Animals; Antioxidants; Apoptosis; Ascorbic Acid; Cadmium; Caspases; Cell Survival; Chromans; DNA Fragmentation; Free Radical Scavengers; Hormones; Immunohistochemistry; Male; Oxidative Stress; Pituitary Gland, Anterior; Prolactin; Rats; Rats, Wistar; Tetrazolium Salts; Thiazoles; Vitamin E

2003
Inhibition of mitochondrial creatine kinase activity by D-2-hydroxyglutaric acid in cerebellum of young rats.
    Neurochemical research, 2003, Volume: 28, Issue:9

    D-2-Hydroxyglutaric aciduria (DHGA) is a neurometabolic disorder biochemically characterized by tissue accumulation and excretion of high amounts of D-2-hydroxyglutaric acid (DGA). Although the affected patients have predominantly severe neurological findings, the underlying mechanisms of brain injury are virtually unknown. In previous studies we have demonstrated that DGA, at concentrations as low as 0.25 mM, significantly decreased creatine kinase activity and other parameters of energy metabolism in cerebral cortex of young rats. In the present study, we investigated the effect of DGA (0.25-5 mM) on total creatine kinase (tCK) activity, as well as on CK activity in cytosolic (Cy-CK) and mitochondrial (Mi-CK) preparations from cerebellum of 30-day-old Wistar rats in order to test whether the inhibitory effect of DGA on CK was tissue specific. We verified that tCK (22% inhibition) and Mi-CK (40% inhibition) activities were moderately inhibited by DGA at concentrations of 2.5 mM and higher, in contrast to Cy-CK, which was not affected by the acid. Kinetic studies revealed that the inhibitory effect of DGA was noncompetitive in relation to phosphocreatine. We also observed that this inhibition was fully prevented by preincubation of the homogenates with reduced glutathione, suggesting that the inhibition of CK activity by DGA is possibly mediated by modification of essential thiol groups of the enzyme. Our present results therefore demonstrate a relatively weak inhibitory effect of DGA on cerebellum Mi-CK activity, as compared to that provoked in cerebral cortex, and may possibly be related to the neuropathology of DHGA, characterized by cerebral cortex abnormalities.

    Topics: Animals; Antioxidants; Ascorbic Acid; Cerebellum; Chromans; Creatine Kinase; Free Radical Scavengers; Glutarates; Glutathione; In Vitro Techniques; Isoenzymes; Mitochondria; NG-Nitroarginine Methyl Ester; Nitric Oxide Synthase; Organ Specificity; Rats; Rats, Wistar; Stereoisomerism

2003
Cooperative interaction between ascorbate and glutathione during mitochondrial impairment in mesencephalic cultures.
    Journal of neurochemistry, 2003, Volume: 86, Issue:6

    A decrease in total glutathione, and aberrant mitochondrial bioenergetics have been implicated in the pathogenesis of Parkinson's disease. Our previous work exemplified the importance of glutathione (GSH) in the protection of mesencephalic neurons exposed to malonate, a reversible inhibitor of mitochondrial succinate dehydrogenase/complex II. Additionally, reactive oxygen species (ROS) generation was an early, contributing event in malonate toxicity. Protection by ascorbate was found to correlate with a stimulated increase in protein-glutathione mixed disulfide (Pr-SSG) levels. The present study further examined ascorbate-glutathione interactions during mitochondrial impairment. Depletion of GSH in mesencephalic cells with buthionine sulfoximine potentiated both the malonate-induced toxicity and generation of ROS as monitored by dichlorofluorescein diacetate (DCF) fluorescence. Ascorbate completely ameliorated the increase in DCF fluorescence and toxicity in normal and GSH-depleted cultures, suggesting that protection by ascorbate was due in part to upstream removal of free radicals. Ascorbate stimulated Pr-SSG formation during mitochondrial impairment in normal and GSH-depleted cultures to a similar extent when expressed as a proportion of total GSH incorporated into mixed disulfides. Malonate increased the efflux of GSH and GSSG over time in cultures treated for 4, 6 or 8 h. The addition of ascorbate to malonate-treated cells prevented the efflux of GSH, attenuated the efflux of GSSG and regulated the intracellular GSSG/GSH ratio. Maintenance of GSSG/GSH with ascorbate plus malonate was accompanied by a stimulation of Pr-SSG formation. These findings indicate that ascorbate contributes to the maintenance of GSSG/GSH status during oxidative stress through scavenging of radical species, attenuation of GSH efflux and redistribution of GSSG to the formation of mixed disulfides. It is speculated that these events are linked by glutaredoxin, an enzyme shown to contain both dehydroascorbate reductase as well as glutathione thioltransferase activities.

    Topics: Animals; Antioxidants; Ascorbic Acid; Buthionine Sulfoximine; Chromans; Disulfides; Fluorescent Dyes; Glutathione; Glutathione Disulfide; Malonates; Mesencephalon; Mitochondria; Oxidative Stress; Parkinson Disease; Rats; Rats, Sprague-Dawley

2003
Urinary excretion of 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone, a ring-fission metabolite of (-)-epicatechin, in rats and its in vitro antioxidant activity.
    Journal of agricultural and food chemistry, 2003, Nov-05, Volume: 51, Issue:23

    There is great interest in the nutritional potential of (-)-epicatechin, a common polyphenolic constituent of many foods and beverages, because of its potent antioxidant capacity. To better evaluate the biological role of (-)-epicatechin, we studied the urinary excretion of 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone, a ring-fission metabolite of (-)-epicatechin by intestinal microflora, in rats as well as its antioxidant activity in vitro. The method for measuring the urinary levels of (-)-epicatechin and 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone was based on the enzymatic hydrolysis of beta-glucuronidase and sulfatase, and was subsequently determined by HPLC coupled to an electrochemical detector. Following administration of (-)-epicatechin at doses of 0, 20, 40, and 80 mumol per rat, (-)-epicatechin and 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone were excreted into the urine within 24 h in a dose-dependent manner. Urinary 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone was mostly in the conjugated form, with a higher ratio of conjugation than (-)-epicatechin. We assessed the relative antioxidant potentials for scavenging radicals in the aqueous phase as expressed in the Trolox equivalent antioxidant capacity (TEAC). The results demonstrated that the degradation of (-)-epicatechin into 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone attenuated the antioxidant ability of the former. However, 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone showed stronger antioxidant activity than l-ascorbic acid. These results led us to suppose that 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone, a microbial metabolite of (-)-epicatechin, circulating in the body may also at least be biologically active in terms of contributing to its combined antioxidant effect.

    Topics: Animals; Antioxidants; Ascorbic Acid; Catechin; Chromans; Chromatography, High Pressure Liquid; Lactones; Male; Rats; Rats, Wistar

2003
Inhibitory activity of flavonoids from Prunus davidiana and other flavonoids on total ROS and hydroxyl radical generation.
    Archives of pharmacal research, 2003, Volume: 26, Issue:10

    Since reactive oxygen species (ROS) and hydroxyl radicals (*OH) play an important role in the pathogenesis of many human degenerative diseases, much attention has focused on the development of safe and effective antioxidants. Preliminary experiments have revealed that the methanol (MeOH) extract of the stem of Prunus davidiana exerts inhibitory/scavenging activities on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, total ROS and peroxynitrites (ONOO-). In the present study, the antioxidant activities of this MeOH extract and the organic solvent-soluble fractions, dichloromethane (CH2Cl2), ethyl acetate (EtOAc), and n-butanol (n-BuOH), and the water layer of P. davidiana stem were evaluated for the potential to inhibit *OH and total ROS generation in kidney homogenates using 2',7'-dichlorodihydrofluorescein diacetate (DCHF-DA), and for the potential to scavenge authentic ONOO-. We also evaluated the inhibitory activity of seven flavonoids isolated from P. davidiana stem, kaempferol, kaempferol 7-O-beta-D-glucoside, (+)-catechin, dihydrokaempferol, hesperetin 5-O-beta-D-glucoside, naringenin and its 7-O-beta-D-glucoside, on the total ROS, *OH and ONOO- systems. For the further elucidation of the structure-inhibitory activity relationship of flavonoids on total ROS and *OH generation, we measured the antioxidant activity of sixteen flavonoids available, including three active flavonoids isolated from P. davidiana, on the total ROS and *OH systems. We found that the inhibitory activity on total ROS generation increases in strength with more numerous hydroxyl groups on their structures. Also, the presence of an ortho-hydroxyl group, whether on the A-ring or B-ring, and a 3-hydroxyl group on the C-ring increased the inhibitory activity on both total ROS and *OH generation.

    Topics: 1-Butanol; Acetates; Animals; Antioxidants; Ascorbic Acid; Biphenyl Compounds; Catechin; Chromans; Drug Evaluation, Preclinical; Flavonoids; Free Radical Scavengers; Hesperidin; Hydrazines; Hydroxyl Radical; Kaempferols; Kidney; Male; Methanol; Methylene Chloride; Penicillamine; Peroxynitrous Acid; Picrates; Plant Extracts; Plant Stems; Prunus; Rats; Rats, Wistar; Reactive Oxygen Species; Structure-Activity Relationship; Water

2003
Diabetes-induced activation of nuclear transcriptional factor in the retina, and its inhibition by antioxidants.
    Free radical research, 2003, Volume: 37, Issue:11

    Oxidative stress is increased in the retina in diabetes, and long-term administration of antioxidants inhibits the development of retinopathy in diabetic rats. The purpose of this study is to determine how diabetes affects the activation of a redox-sensitive nuclear transcriptional factor in the retina, NF-kappaB, and its inhibition by antioxidants. Alloxan diabetic rats were assigned to receive standard diet or the diet supplemented with multiple antioxidants, including ascorbic acid, Trolox, dl alpha-tocopherol acetate, N-acetyl cysteine, beta-carotene, and selenium for up to 14 months. NF-kappaB activation, oxidative stress and nitric oxides were measured in the retina at 2, 8 and 14 months of diabetes. Retinal NF-kappaB was activated by about 60% at two months after induction of diabetes, remained activated for up to 14 months of diabetes, and the duration of diabetes had no effect on the intensity of NF-kappaB activation. Similarly, oxidative stress and nitric oxides were elevated by over 50% in the retina of rats diabetic for 14 months, and nitrotyrosine levels were elevated by over two folds. Administration of the antioxidants to the rats for the entire duration of diabetes inhibited activation of NF-kappaB and elevations in oxidative stress, nitric oxides and nitrotyrosine formation without ameliorating the severity of hyperglycemia. These in vivo results were confirmed by in vitro studies showing that high glucose activates NF-kappaB and elevates NO and lipid peroxides in both retinal endothelial cells and pericytes that can be inhibited by antioxidants. Thus, the results suggest that the activation of retinal NF-KB in diabetes is an early event in the development of retinopathy, and it remains active when the retinal capillary cell death is accelerating, and histopathology is developing. Beneficial effects of antioxidants on the development of diabetic retinopathy might involve inhibition of NF-kappaB activation and its downstream pathways in the retina.

    Topics: alpha-Tocopherol; Animals; Antioxidants; Ascorbic Acid; beta Carotene; Chromans; Cysteine; Diabetes Mellitus, Experimental; Diabetic Retinopathy; Glucose; Lipid Peroxides; Male; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Rats; Rats, Sprague-Dawley; Retina; Selenium; Tyrosine

2003
In vitro antioxidant activity of 2,5,7,8-tetramethyl-2-(2'-carboxyethyl)-6-hydroxychroman (alpha-CEHC), a vitamin E metabolite.
    Free radical research, 2002, Volume: 36, Issue:8

    2,5,7,8-tetramethyl-2-(2'-carboxyethyl)-6-hydroxychroman (alpha-CEHC) has been identified as a major water-soluble metabolite of vitamin E, which circulates in the blood and is excreted with the urine. The aim of this study was to assess the antioxidant activity of alpha-CEHC using several methods with different prooxidant challenges. In the Oxygen Radical Absorbance Capacity assay, a fluorescent protein acts as a marker for oxidative damage induced by peroxyl radicals. In the Trolox Equivalent Antioxidant Capacity (TEAC) assay, a stable free radical, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid (ABTS.+) is reduced directly by antioxidants. Scavenging properties vs. reactive nitrogen species were studied measuring the effects on tyrosine nitration after reaction with peroxynitrite. Trolox, alpha-tocopherol, ascorbic acid, and (-)-epicatechin were simultaneously tested in order to compare their antioxidant activities. In all mentioned systems, alpha-CEHC exhibited antioxidant properties similar to those of Trolox. We conclude that alpha-CEHC is a molecule with good antioxidant activity, having the advantage over Trolox of being a naturally occurring compound. These properties might be useful for research or industrial purposes.

    Topics: Antioxidants; Ascorbic Acid; Chromans; Dose-Response Relationship, Drug; Humans; In Vitro Techniques; Models, Chemical; Nitrogen; Oxygen; Peroxynitrous Acid; Propionates; Spectrometry, Fluorescence; Time Factors; Tyrosine; Vitamin E

2002
Inhibition of lipid peroxidation by S-nitrosoglutathione and copper.
    Free radical research, 2002, Volume: 36, Issue:10

    The antioxidant properties of S-nitrosoglutathione, a nitric oxide-derived product were studied in different experimental systems. By using the crocin bleaching test, S-nitrosoglutathione, in the presence of copper ions, shows an antioxidant capacity about six times higher than that of Trolox c and referable to the interception of peroxyl radicals by nitric oxide. Copper alone shows a modest inhibitory action, which is about seven times lower than that of Trolox c. S-nitrosoglutathione prevents lipid peroxidation induced by the well-known Fe2+/ascorbate system (IC50 = 450 microM) and the inhibitory effect is strongly reinforced by the presence of copper ions (IC50 = 6.5 microM). In addition, cumene hydroperoxide-induced lipid peroxidation is markedly decreased by S-nitrosoglutathione, provided that copper ions, maintained reduced by ascorbate, are present. Decomposition of S-nitrosoglutathione through metal catalysis and/or the presence of reducing agents and the consequent release of nitric oxide are of crucial importance for eliciting the antioxidant power. In this way, copper ions and/or reducing species with low antioxidant potency are able to promote the formation of an extremely strong antioxidant species such as nitric oxide.

    Topics: Animals; Antioxidants; Ascorbic Acid; Benzene Derivatives; Carotenoids; Chromans; Copper; Ferrous Compounds; Lipid Peroxidation; Microsomes, Liver; Nitric Oxide; Peroxides; Rats; Reducing Agents; S-Nitrosoglutathione; Thiobarbituric Acid Reactive Substances

2002
Radical scavengers from the entomogenous deuteromycete Beauveria amorpha.
    Planta medica, 2002, Volume: 68, Issue:1

    The culture medium of Beauveria amorpha Hohn. exhibited strong radical scavenging properties. Activity-directed fractionation assisted by a radical scavenging assay on TLC afforded two active compounds which were identified as 6-methyl-2,4-dihydroxyphenyl 4-O-methyl-beta-D-glucopyranoside (1) and (-)-terredionol (2). Compound 1 displays radical scavenging and lipid peroxidation inhibitory activity comparable to those of TroloxC, ascorbic acid or quercetin.

    Topics: Animals; Antioxidants; Ascorbic Acid; Chromans; Cyclohexanones; Free Radical Scavengers; Lipid Peroxidation; Lipid Peroxides; Magnetic Resonance Spectroscopy; Mice; Mitosporic Fungi; Monosaccharides; PC12 Cells; Quercetin; Rats; Vitamin E

2002
CX-659S: a novel diaminouracil derivative that has antioxidative and acute anti-inflammatory activities.
    European journal of pharmacology, 2002, Mar-08, Volume: 438, Issue:3

    We investigated the antioxidative activities and the effects on acute inflammation in mice of a novel diaminouracil derivative, CX-659S ((S)-6-amino-5-(6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxamido)-3-methyl-1-phenyl-2,4(1H,3H)-pyrimidinedione). CX-659S showed potent scavenging activities against the hydroxyl radical and peroxynitrite and inhibited lipid peroxidation in rat brain homogenates in vitro. Topically applied CX-659S dose-dependently inhibited arachidonic acid- and 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced ear edema in mice. Consistent with its antioxidative properties in vitro, CX-659S dramatically attenuated the accumulation of lipid peroxides in the mouse ear elicited by repeated application of TPA. Previously, we reported the effectiveness of CX-659S against contact hypersensitivity reactions in both mouse and guinea pig models. These present results further suggest the therapeutic potential of CX-659S for acute skin inflammation that may involve oxidative tissue damage.

    Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Arachidonic Acid; Ascorbic Acid; Chromans; Dose-Response Relationship, Drug; Ear; Edema; Lipid Peroxidation; Male; Mice; Mice, Inbred ICR; Oxidation-Reduction; Peroxynitrous Acid; Rats; Rats, Sprague-Dawley; Tetradecanoylphorbol Acetate; Uracil

2002
The effect of antioxidants on glycated albumin-induced cytotoxicity in bovine retinal pericytes.
    Biochemical and biophysical research communications, 2002, Apr-12, Volume: 292, Issue:4

    Loss of retinal pericytes is the initial deficit in the early stage of diabetic retinopathy. Glycated albumin (GA) forms under hyperglycemic conditions and exists in the retinal blood vessels of diabetic patients with retinopathy. In this study, using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) reduction test, we investigated whether GA induces cytotoxicity in cultured bovine retinal pericytes, and whether the antioxidants, l-ascorbic acid, Trolox, and probucol, provide any protection from GA-mediated cytotoxicity. GA induced pericyte death in a dose-dependent manner. With increasing time, GA-induced cytotoxicity also increased despite no strict time dependence. Furthermore, this cell death was found to be mediated both by apoptosis, which was confirmed by apoptosis-specific fluorescent staining of nuclei and cell membranes, and mitochondrial damage, as elucidated by electron microscopy. All three antioxidants used in this study partially protected against GA-induced pericyte death, suggesting that oxidative stress plays a role in GA-induced pericyte death. The results indicate that GA induces cell death in cultured bovine retinal pericytes, and that certain antioxidants may reduce this cytotoxicity.

    Topics: Animals; Antioxidants; Ascorbic Acid; Cattle; Cell Death; Cell Survival; Cells, Cultured; Chromans; Cytoprotection; Diabetic Retinopathy; Dose-Response Relationship, Drug; Glycated Serum Albumin; Glycation End Products, Advanced; Mitochondria; Oxidative Stress; Pericytes; Probucol; Retina; Retinal Vessels; Serum Albumin; Tetrazolium Salts; Thiazoles; Time Factors

2002
An investigation into the mechanisms mediating plasma lipoprotein-potentiated beta-amyloid fibrillogenesis.
    FEBS letters, 2002, May-08, Volume: 518, Issue:1-3

    The toxicity of the beta-amyloid (Abeta) peptide of Alzheimer's disease may relate to its polymerisation state (i.e. fibril content). We have shown previously that plasma lipoproteins, particularly when oxidised, greatly enhance Abeta polymerisation. In the present study the nature of the interactions between both native and oxidised lipoproteins and Abeta1-40 was investigated employing various chemical treatments. The addition of ascorbic acid or the vitamin E analogue, trolox, to lipoprotein/Abeta coincubations failed to inhibit Abeta fibrillogenesis, as did the treatment of lipoproteins with the aldehyde reductant, sodium borohydride. The putative lipid peroxide-derived aldehyde scavenger, aminoguanidine, however, inhibited Abeta-oxidised lipoprotein-potentiated polymerisation, but in a manner consistent with an antioxidant action for the drug. Lipoprotein treatment with the reactive aldehyde 4-hydroxy-2-trans-nonenal enhanced Abeta polymerisation in a concentration-dependent fashion. Incubation of Abeta with lipoprotein fractions from which the apoprotein components had been removed resulted in extents of polymerisation comparable to those observed with Abeta alone. These data indicate that the apoprotein components of plasma lipoproteins play a key role in promoting Abeta polymerisation, possibly via interactions with aldehydes.

    Topics: Aldehydes; Alzheimer Disease; Amyloid beta-Peptides; Antioxidants; Apolipoproteins; Ascorbic Acid; Biopolymers; Borohydrides; Chromans; Guanidines; Humans; Kinetics; Lipoproteins; Oxidation-Reduction; Peptide Fragments

2002
Assessment of antioxidant activity by using different in vitro methods.
    Free radical research, 2002, Volume: 36, Issue:2

    In this study, six common tests for measuring antioxidant activity were evaluated by comparing four antioxidants and applying them to beverages (tea and juices): Trolox equivalent antioxidant capacity assay (TEAC I-III assay), Total radical-trapping antioxidant parameter assay (TRAP assay), 2,2-diphenyl-l-picrylhydrazyl assay (DPPH assay), N,N-dimethyl-p-phenylendiamine assay (DMPD assay), Photochemiluminescence assay (PCL assay) and Ferric reducing ability of plasma assay (FRAP assay). The antioxidants included gallic acid representing the group of polyphenols, uric acid as the main antioxidant in human plasma, ascorbic acid as a vitamin widely spread in fruits and Trolox as water soluble vitamin E analogue. The six methods presented can be divided into two groups depending on the oxidising reagent. Five methods use organic radical producers (TEAC I-III, TRAP, DPPH, DMPD, PCL) and one method works with metal ions for oxidation (FRAP). Another difference between these tests is the reaction procedure. Three assays use the delay in oxidation and determine the lag phase as parameter for the antioxidant activity (TEAC I, TRAP, PCL). They determine the delay of radical generation as well as the ability to scavenge the radical. In contrast, the assays TEAC II and III, DPPH, DMPD and FRAP analyse the ability to reduce the radical cation (TEAC II and III, DPPH, DMPD) or the ferric ion (FRAP). The three tests acting by radical reduction use preformed radicals and determine the decrease in absorbance while the FRAP assay measures the formed ferrous ions by increased absorbance. Gallic acid was the strongest antioxidant in all tests with exception of the DMPD assay. In contrast, uric acid and ascorbic acid showed low activity in some assays. Most of the assays determine the antioxidant activity in the micromolar range needing minutes to hours. Only one assay (PCL) is able to analyse the antioxidant activity in the nanomolar range. Black currant juice showed highest antioxidant activity in all tests compared to tea, apple juice and tomato juice. Despite these differences, results of these in vitro assays give an idea of the protective efficacy of secondary plant products. It is strongly recommended to use at least two methods due to the differences between the test systems investigated.

    Topics: Antioxidants; Ascorbic Acid; Beverages; Chemistry Techniques, Analytical; Chromans; Fluorescence; Fruit; Gallic Acid; Reproducibility of Results; Sensitivity and Specificity; Tea; Uric Acid

2002
The antioxidant activity of regularly consumed fruit and vegetables reflects their phenolic and vitamin C composition.
    Free radical research, 2002, Volume: 36, Issue:2

    Recent studies are emphasising the importance and putative modes of action of specific flavonoids as bioactive components of the diet in in vivo and in vitro models. Thus, it is important to have a clear idea of the major phenolic families of which fruit and vegetables are comprised and the levels contained therein. Regularly consumed fruit and vegetables of mixed varieties available on the UK market were analysed for the composition of the major individual phenolic components. The total phenolic content (applying the Folin assay) and the vitamin C levels were also determined. The antioxidant capacities of aqueous/methanolic extracts were comparatively assessed using the TEAC (Trolox Equivalent Antioxidant Capacity), the FRAP (Ferric Reducing Ability of Plasma) and ORAC (Oxygen Radical Absorbance Capacity) assays, which comprise contributions from polyphenols, simple phenols and the ascorbate component. The results were calculated in terms of 100 g fresh weight (FW) uncooked portion sizes. Fruit and vegetables rich in anthocyanins (e.g. strawberry, raspberry and red plum) demonstrated the highest antioxidant activities, followed by those rich in flavanones (e.g. orange and grapefruit) and flavonols (e.g. onion, leek, spinach and green cabbage), while the hydroxycinnamate-rich fruit (e.g. apple, tomato, pear and peach) consistently elicited the lower antioxidant activities. The TEAC, FRAP and ORAC values for each extract were relatively similar and well-correlated with the total phenolic and vitamin C contents. The antioxidant activities (TEAC) in terms of 100 g FW uncooked portion size were in the order: strawberry>> raspberry = red plum >> red cabbage >>>grapefruit = orange > spinach > broccoli > green grape approximately/= onion > green cabbage > pea > apple > cauliflower tomato approximately/= peach=leek > banana approximately/= lettuce.

    Topics: Anthocyanins; Antioxidants; Ascorbic Acid; Chromans; Chromatography, High Pressure Liquid; Coumaric Acids; Flavonoids; Fruit; Oxidation-Reduction; Phenols; Reactive Oxygen Species; Spectrophotometry, Ultraviolet; Vegetables

2002
Vitamin C equivalent antioxidant capacity (VCEAC) of phenolic phytochemicals.
    Journal of agricultural and food chemistry, 2002, Jun-19, Volume: 50, Issue:13

    To express the antioxidant capacity of plant foods in a more familiar and easily understood manner (equivalent to vitamin C mg/100 g), two stable radical species, ABTS(*)(-) and DPPH(*), commonly used for antioxidant activity measurements, were employed independently to evaluate their efficacies using apple polyphenolic extracts and seven polyphenolic standards including synthetic Trolox. Their antioxidant activities were expressed as vitamin C equivalent antioxidant capacity (VCEAC) in mg/100 g apple or mg/100 mL of the reference chemical compounds in 10 and 30 min using the ABTS(*)(-) and DPPH(*) scavenging assays, respectively. The antioxidant capacity of Gala apples and seven phenolic standards, determined by both ABTS(*)(-) and DPPH(*) scavenging assays, showed a dose-response of the first-order. Fresh Gala apples had a VCEAC of 205.4 +/- 5.6 mg/100 g using the ABTS assay, and the relative VCEACs of phenolic standards were as follows: gallic acid > quercetin > epicatechin > catechin > vitamin C > rutin > chlorogenic acid > Trolox. With the DPPH radical assay, the VCEAC of fresh Gala apples was 136.0 +/- 6.6 mg/100 g, and the relative VCEACs of seven phenolic standards were, in decreasing order, as follows: gallic acid > quercetin > epicatechin > catechin > or = vitamin C > Trolox > rutin > chlorogenic acid. Because the ABTS assay can be used in both organic and aqueous solvent systems, employs a specific absorbance at a wavelength remote from the visible region, and requires a short reaction time, it is a more desirable method than the DPPH assay. Therefore, it is recommended that antioxidant capacity be expressed as vitamin C mg/100 g equivalent (VCEAC) using the ABTS assay.

    Topics: Antioxidants; Ascorbic Acid; Benzothiazoles; Biphenyl Compounds; Catechin; Chlorogenic Acid; Chromans; Free Radical Scavengers; Free Radicals; Gallic Acid; Kinetics; Malus; Oxidation-Reduction; Phenols; Picrates; Plant Extracts; Plants, Edible; Quercetin; Rutin; Sulfonic Acids

2002
A reassessment of the peroxynitrite scavenging activity of uric acid.
    Annals of the New York Academy of Sciences, 2002, Volume: 962

    Peroxynitrite is implicated in numerous human diseases. Hence, there is considerable interest in potential therapeutic peroxynitrite scavengers. It has been claimed that uric acid is a powerful peroxynitrite scavenger. We previously observed that uric acid is a powerful inhibitor of tyrosine nitration induced by peroxynitrite, but fails to prevent alpha(1)-antiproteinase (alpha(1)-AP) inactivation induced by peroxynitrite. However, the reactivity of peroxynitrite is significantly modified by bicarbonate and this has not been considered in evaluating the scavenging activity of uric acid and other endogenous antioxidant compounds. In the presence of bicarbonate (25 mM), the ability of uric acid, ascorbate, Trolox, and GSH to inhibit peroxynitrite-mediated tyrosine and guanine nitration is decreased. Protection against peroxynitrite-mediated alpha(1)-AP inactivation is also decreased by ascorbate, Trolox, and GSH, but it is enhanced by uric acid. Bicarbonate also inhibits the ability of these compounds to prevent peroxynitrite-mediated ABTS radical cation formation. However, the abilities of these antioxidants to prevent peroxynitrite-mediated bleaching of pyrogallol red are enhanced by bicarbonate. These results show that physiologic concentrations of bicarbonate substantially modify the ability of uric acid to prevent peroxynitrite-mediated reactions. This study highlights the need to use several different assays in the presence of physiologically relevant concentrations of bicarbonate when assessing compounds for peroxynitrite scavenging, in order to avoid misleading results.

    Topics: alpha 1-Antitrypsin; Antioxidants; Ascorbic Acid; Benzothiazoles; Bicarbonates; Chromans; Coloring Agents; Glutathione; Guanine; Humans; Indicators and Reagents; Peroxynitrous Acid; Pyrogallol; Reactive Nitrogen Species; Serine Proteinase Inhibitors; Sulfonic Acids; Tyrosine; Uric Acid

2002
Intracellular antioxidant activity is necessary to modulate the macrophage response to endotoxin.
    Shock (Augusta, Ga.), 2002, Volume: 18, Issue:1

    The tissue-fixed macrophage (Mphi) is a key cell in the coordination of the excessive systemic immunoinflammatory response underlying the adult respiratory distress syndrome (ARDS). Macrophage-generated reactive oxygen intermediates (ROIs) are involved in both tissue destruction via lipid peroxidation and in the activation of these inflammatory cells. It is unclear whether oxidant-induced activation involves an extracellular effect and membrane destabilization or occurs through intracellular alteration of the redox state and direct involvement as second messengers. In this study, we compare the differential effects of known intracellular vs. extracellular antioxidants on the Mphi response to endotoxin. Rabbit alveolar Mphi were obtained by bronchoalveolar lavage and exposed to either the extracellular antioxidants [vitamin C (VC) (10-1000 microM), Trolox (100-1000 microM, superoxide dismutase (SOD) (10-500 microM))] or the intracellular antioxidants [N-acetylcysteine (NAC) (0.1-10 mM) or butylated hydroxyanisole (BHA) (10-200 microM)] for 1 h. Cells were subsequently stimulated with lipopolysaccharide at 10 ng/mL. After 18 h, supernatants were analyzed for tumor necrosis factor (TNF) and F2 isoprostane (F2ISP) production and cellular monolayers for procoagulant activity (PCA). A dose response inhibition of both TNF and PCA production was demonstrated after both NAC and BHA pretreatment but not with VC, Trolox, or SOD. In addition, northern blots revealed inhibition of TNF mRNA production by both NAC and BHA. F2ISP, a marker of membrane lipid peroxidation, was inhibited by BHA and Trolox but not NAC, VC, or SOD. In conclusion, antioxidants that are incorporated intracellularly are expected to be beneficial in the treatment of excessive inflammatory responses through the interruption of redox dependent signal transduction pathways and subsequent modulation of the Mphi proinflammatory response.

    Topics: Acetylcysteine; Animals; Antioxidants; Ascorbic Acid; Blood Coagulation Factors; Butylated Hydroxyanisole; Cells, Cultured; Chromans; Endotoxins; F2-Isoprostanes; Inflammation; Macrophages, Alveolar; Male; Rabbits; Reactive Oxygen Species; Respiratory Distress Syndrome; Signal Transduction; Superoxide Dismutase; Tumor Necrosis Factor-alpha

2002
Lucigenin chemiluminescence in human seminal plasma.
    Free radical research, 2001, Volume: 34, Issue:2

    Seminal plasma protects spermatozoa from the detrimental effects of reactive oxygen species such as hydrogen peroxide. We investigated the lucigenin-dependent chemiluminescence in cell-free seminal plasma from andrological patients. The seminal plasma was separated from cells by centrifugation. In all seminal plasmas studied lucigenin-dependent chemiluminescence (LCL) was detected. The LCL showed a strong pH-dependence. The signal was stable if samples were stored at +4 degrees C for up to 4 days or up to 8 days at -80 degrees C. Filtration of the samples (0.45 and 0.22 microm pore size) did not lower their luminescence. The addition of superoxide dismutase (SOD) and ascorbic acid oxidase (AAO) lowered LCL nearly to baseline values while trolox and desferal showed moderate effect, whereas allopurinol had no effect. Electron paramagnetic resonance spectroscopy demonstrated ascorbyl radicals in seminal plasma. Physiological concentrations of ascorbic acid yielded SOD-inhibitable lucigenin-chemiluminescence. The nitroblue-tetrazolium assay showed that ascorbic acid in buffer solution produced formazan. Superoxide-anion radicals were not detected in seminal plasma by the spin-trap DEPMPO due to their low steady state concentration. It is concluded that in seminal plasma ascorbate reacts with molecular oxygen yielding ascorbyl radicals and superoxide anion. If lucigenin is added to seminal plasma, reducing substances present, such as ascorbate, reduce lucigenin to the corresponding radical; this radical reacts with molecular oxygen and also forms O2-. So LCL in human seminal plasma results from the autoxidation of ascorbate and the oxidation of the reduced lucigenin. While the physiological relevance of the former mechanism is unknown, the latter is an artifact.

    Topics: Acridines; Adult; Allopurinol; Antioxidants; Ascorbate Oxidase; Ascorbic Acid; Chromans; Deferoxamine; Electron Spin Resonance Spectroscopy; Free Radicals; Humans; Hydrogen-Ion Concentration; Infertility, Male; Luminescent Measurements; Male; Oxidants; Oxidation-Reduction; Oxygen; Oxygen Consumption; Reactive Oxygen Species; Semen; Semen Preservation; Superoxide Dismutase; Temperature

2001
The binding of oxidized low density lipoprotein (ox-LDL) to ox-LDL receptor-1 reduces the intracellular concentration of nitric oxide in endothelial cells through an increased production of superoxide.
    The Journal of biological chemistry, 2001, Apr-27, Volume: 276, Issue:17

    Oxidized low density lipoprotein (ox-LDL) has been suggested to affect endothelium-dependent vascular tone through a decreased biological activity of endothelium-derived nitric oxide (NO). Oxidative inactivation of NO is regarded as an important cause of its decreased biological activity, and in this context superoxide (O(2)) is known to inactivate NO in a chemical reaction during which peroxynitrite is formed. In this study we examined the effect of ox-LDL on the intracellular NO concentration in bovine aortic endothelial cells and whether this effect is influenced by ox-LDL binding to the endothelial receptor lectin-like ox-LDL receptor-1 (LOX-1) through the formation of reactive oxygen species and in particular of O(2). ox-LDL induced a significant dose-dependent decrease in intracellular NO concentration both in basal and stimulated conditions after less than 1 min of incubation with bovine aortic endothelial cells (p < 0.01). In the same experimental conditions ox-LDL also induced O(2) generation (p < 0.001). In the presence of radical scavengers and anti-LOX-1 monoclonal antibody, O(2) formation induced by ox-LDL was reduced (p < 0.001) with a contemporary rise in intracellular NO concentration (p < 0.001). ox-LDL did not significantly modify the ability of endothelial nitric oxide synthase to metabolize l-arginine to l-citrulline. The results of this study show that one of the pathophysiological consequences of ox-LDL binding to LOX-1 may be the inactivation of NO through an increased cellular production of O(2).

    Topics: Allopurinol; Animals; Antibodies, Monoclonal; Antioxidants; Aorta; Ascorbic Acid; Aspirin; Bradykinin; Catecholamines; Cattle; Cells, Cultured; CHO Cells; Chromans; Cricetinae; Dose-Response Relationship, Drug; Endothelium, Vascular; Enzyme Inhibitors; Flow Cytometry; Free Radical Scavengers; Hemostatics; Humans; Imidazolines; Lipoproteins, LDL; Mice; Nitric Oxide; omega-N-Methylarginine; Oxygen; Probucol; Protein Binding; Reactive Oxygen Species; Receptors, LDL; Receptors, Oxidized LDL; Scavenger Receptors, Class E; Superoxides; Thrombin; Time Factors

2001
Antioxidants inhibit JNK and p38 MAPK activation but not ERK 1/2 activation by angiotensin II in rat aortic smooth muscle cells.
    Hypertension research : official journal of the Japanese Society of Hypertension, 2001, Volume: 24, Issue:3

    Angiotensin II (Ang II) induces vascular smooth muscle cell (VSMC) hypertrophy, which results in several cardiovascular diseases. Ang II-induced cellular events have been mediated, in part, by reactive oxygen species (ROS) which also involve activation of mitogen-activated protein (MAP) kinases. Although it has been proposed that the therapeutic administration of antioxidants is useful for vascular diseases, the precise mechanisms which regulate ROS-sensitive signaling events have not been well characterized. Thus, we hypothesized that antioxidants may affect ROS-mediated MAP kinases activation induced by Ang II. The present findings showed that Ang II stimulated rapid and significant activation of ERK 1/2, JNK and p38 MAPK in cultured rat aortic smooth muscle cells (RASMC). Ang II-induced ERK 1/2 activation was not affected by all antioxidants examined, whereas JNK was sensitive to all antioxidants. In contrast, p38 MAPK activation was inhibited by DPI and ascorbic acid concentration-dependently, but by NAC only at high concentration. DETC and Trolox C had no effects on p38 MAPK activation by Ang II. We further examined the effects of antioxidants on Ang II-induced increases in oxygen consumption as an index of ROS generation in RASMC. DPI strongly inhibited Ang II-induced increases in oxygen consumption. DETC also inhibited Ang II-induced oxygen consumption, whereas ascorbic acid markedly augmented it. These findings suggest that the inhibitory effects of antioxidants on MAP kinases activation in VSMC are attributable, in part, to their modulating effects on ROS generation by Ang II in VSMC. Thus, inhibition of MAP kinases by antioxidants may imply their usefulness for relief of cardiovascular diseases.

    Topics: Acetylcysteine; Angiotensin II; Animals; Antioxidants; Aorta, Thoracic; Ascorbic Acid; Cells, Cultured; Chelating Agents; Chromans; Ditiocarb; Dose-Response Relationship, Drug; Enzyme Inhibitors; Free Radical Scavengers; JNK Mitogen-Activated Protein Kinases; Male; MAP Kinase Kinase 4; MAP Kinase Signaling System; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinase Kinases; Mitogen-Activated Protein Kinases; Muscle, Smooth, Vascular; Onium Compounds; Oxygen Consumption; p38 Mitogen-Activated Protein Kinases; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Vasoconstrictor Agents

2001
Hydrogen peroxide removal and glutathione mixed disulfide formation during metabolic inhibition in mesencephalic cultures.
    Journal of neurochemistry, 2001, Volume: 77, Issue:6

    Compromised mitochondrial energy metabolism and oxidative stress have been associated with the pathophysiology of Parkinson's disease. Our previous experiments exemplified the importance of GSH in the protection of neurons exposed to malonate, a reversible inhibitor of mitochondrial succinate dehydrogenase/complex II. This study further defines the role of oxidative stress during energy inhibition and begins to unravel the mechanisms by which GSH and other antioxidants may contribute to cell survival. Treatment of mesencephalic cultures with 10 microM buthionine sulfoximine for 24 h depleted total GSH by 60%, whereas 3 h exposure to 5 mM 3-amino-1,2,4-triazole irreversibly inactivated catalase activity by 90%. Treatment of GSH-depleted cells with malonate (40 mM) for 6, 12 or 24 h both potentiated and accelerated the time course of malonate toxicity, however, inhibition of catalase had no effect. In contrast, concomitant treatment with buthionine sulfoximine plus 3-amino-1,2,4-triazole in the presence of malonate significantly potentiated toxicity over that observed with malonate plus either inhibitor alone. Consistent with these findings, GSH depletion enhanced malonate-induced reactive oxygen species generation prior to the onset of toxicity. These findings demonstrate that early generation of reactive oxygen species during mitochondrial inhibition contributes to cell damage and that GSH serves as a first line of defense in its removal. Pre-treatment of cultures with 400 microM ascorbate protected completely against malonate toxicity (50 mM, 12 h), whereas treatment with 1 mM Trolox provided partial protection. Protein-GSH mixed disulfide formation during oxidative stress has been suggested to either protect vulnerable protein thiols or conversely to contribute to toxicity. Malonate exposure (50 mM) for 12 h resulted in a modest increase in mixed disulfide formation. However, exposure to the protective combination of ascorbate plus malonate increased membrane bound protein-GSH mixed disulfides three-fold. Mixed disulfide levels returned to baseline by 72 h of recovery indicating the reversible nature of this formation. These results demonstrate an early role for oxidative events during mitochondrial impairment and stress the importance of the glutathione system for removal of reactive oxygen species. Catalase may serve as a secondary defense as the glutathione system becomes limiting. These findings also suggest that protein-GSH mixed disulfide formatio

    Topics: Amitrole; Animals; Antioxidants; Ascorbic Acid; Catalase; Cells, Cultured; Chromans; Energy Metabolism; Enzyme Inhibitors; Glutathione Disulfide; Hydrogen Peroxide; Malonates; Mesencephalon; Neurons; Oxidative Stress; Parkinson Disease; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species

2001
Abnormalities of retinal metabolism in diabetes and experimental galactosemia. VII. Effect of long-term administration of antioxidants on the development of retinopathy.
    Diabetes, 2001, Volume: 50, Issue:8

    Antioxidants were administered to diabetic rats and experimentally galactosemic rats to evaluate the ability of these agents to inhibit the development of diabetic retinopathy. Alloxan diabetic rats and nondiabetic rats that were fed 30% galactose randomly received standard diets or the diets supplemented with ascorbic acid and alpha-tocopherol (vitamins C+E diet) or a more comprehensive mixture of antioxidants (multi-antioxidant diet), including Trolox, alpha-tocopherol, N-acetyl cysteine, ascorbic acid, beta-carotene, and selenium. Diabetes or galactose feeding of at least 12 months resulted in pericyte loss, acellular capillaries, and basement membrane thickening. Compared with diabetic controls, the development of acellular capillaries was inhibited by 50% (P < 0.05) in diabetic rats that received supplemental vitamins C+E, and the number of pericyte ghosts tended to be reduced. The vitamins C+E supplement had no beneficial effect in galactosemic rats, but these rats consumed only approximately half as much of the antioxidants as the diabetic rats. The multi-antioxidant diet significantly inhibited ( approximately 55-65%) formation of both pericyte ghosts and acellular capillaries in diabetic rats and galactosemic rats (P < 0.05 vs. controls), without affecting the severity of hyperglycemia. Parameters of retinal oxidative stress, protein kinase C activity, and nitric oxides remained elevated for at least 1 year of hyperglycemia, and these abnormalities were normalized by multi-antioxidant therapy. Thus, long-term administration of antioxidants can inhibit the development of the early stages of diabetic retinopathy, and the mechanism by which this action occurs warrants further investigation. Supplementation with antioxidants can offer an achievable and inexpensive adjunct therapy to help inhibit the development of retinopathy in diabetes.

    Topics: Acetylcysteine; Animals; Antioxidants; Ascorbic Acid; beta Carotene; Chromans; Diabetes Mellitus, Experimental; Diabetic Retinopathy; Diet; Dietary Supplements; Galactosemias; Male; Pericytes; Rats; Rats, Sprague-Dawley; Retina; Retinal Diseases; Selenium; Vitamin E

2001
The 'pivotal antioxidant' hypothesis for the role of flavonoids in their reduction of HO* radical-induced damage on DNA.
    Redox report : communications in free radical research, 2001, Volume: 6, Issue:3

    Topics: Antioxidants; Ascorbic Acid; Catechin; Chromans; DNA Damage; DNA, Bacterial; Flavonoids; Gamma Rays; Glutathione; Hydroxyl Radical; Models, Biological; Oxidants; Oxidation-Reduction

2001
Astaxanthin and peridinin inhibit oxidative damage in Fe(2+)-loaded liposomes: scavenging oxyradicals or changing membrane permeability?
    Biochemical and biophysical research communications, 2001, Oct-19, Volume: 288, Issue:1

    Astaxanthin and peridinin, two typical carotenoids of marine microalgae, and lycopene were incorporated in phosphatidylcholine multilamellar liposomes and tested as inhibitors of lipid oxidation. Contrarily to peridinin results, astaxanthin strongly reduced lipid damage when the lipoperoxidation promoters-H(2)O(2), tert-butyl hydroperoxide (t-ButOOH) or ascorbate-and Fe(2+):EDTA were added simultaneously to the liposomes. In order to check if the antioxidant activity of carotenoids was also related to their effect on membrane permeability, the peroxidation processes were initiated by adding the promoters to Fe(2+)-loaded liposomes (encapsulated in the inner aqueous solution). Despite that the rigidifying effect of carotenoids in membranes was not directly measured here, peridinin probably has decreased membrane permeability to initiators (t-ButOOH > ascorbate > H(2)O(2)) since its incorporation limited oxidative damage on iron-liposomes. On the other hand, the antioxidant activity of astaxanthin in iron-containing vesicles might be derived from its known rigidifying effect and the inherent scavenging ability.

    Topics: Antioxidants; Ascorbic Acid; beta Carotene; Carotenoids; Cell Membrane Permeability; Chromans; Free Radical Scavengers; Hydrogen Peroxide; Iron; Lipid Peroxidation; Liposomes; Models, Biological; Oxidative Stress; Reactive Oxygen Species; tert-Butylhydroperoxide; Thiobarbituric Acid Reactive Substances; Xanthophylls

2001
Bioflavonoid rescue of ascorbate at a membrane interface.
    Journal of bioenergetics and biomembranes, 2001, Volume: 33, Issue:4

    In aqueous solution, ascorbate potently prevents bleaching of cytochrome c on exposure to excess H2O2 or t-butyl hydroperoxide. Ascorbate failed to protect cytochrome c in the presence of liposomes of mitochondrial membranelike composition. Like the redox mediator N,N,N,'N'-tetramethyl-p-phenylenediamine (TMPD), however, the bioflavonoids epicatechin and quercetin restored the protection afforded by ascorbate in the presence of liposomes and gave further protection. The quercetin glycoside, rutin, was much less effective, as was the vitamin E analog Trolox. In the presence of liposomes, quercetin alone was relatively ineffective, but cooperated with ascorbate to extend protection synergistically. The results bear specific implications in antioxidant protection of cytochrome c and in moderation of its hydroperoxidase activities in biological membranes. The data also reveal a situation where ascorbate is without effect except in the presence of a bioflavonoid, and substantiate a possibly vital role for certain bioflavonoids in mediating electron transfer from ascorbate into a hydrophobic environment.

    Topics: Animals; Antioxidants; Ascorbic Acid; Catechin; Chromans; Cytochrome c Group; Drug Synergism; Flavonoids; Kinetics; Liposomes; Oxidation-Reduction; Peroxides; Quercetin; Rutin; Spectrum Analysis

2001
Effects of antioxidants on induction of apoptosis in bursal cells of Fabricius during in vitro cultivation.
    The Journal of veterinary medical science, 2000, Volume: 62, Issue:1

    After physically disrupting cell contacts, apoptosis of bursal cells of Fabricius was induced during in vitro cultivation. The percentage of apoptotic cells increased with incubation time and approximately 70% cells represented apoptosis after 6 hr of incubation. The induction of apoptosis was significantly inhibited by treatment of the cells with ascorbic acid (vitamin C), but not with trolox, a vitamin E analog. An intense DNA ladder pattern was shown at 6 hr post-isolation, which is a biochemical hallmark of apoptosis. Treatment of the cells with ascorbic acid inhibited the DNA fragmentation, but trolox did not. To monitor the intracellular production of reactive oxygen species (ROSs), the intensity of fluorescence emitted from DCFH-DA was measured. The intensity of fluorescence from cells incubated for 0.5-2 hr was approximately 2-fold higher than that from cells at 0 hr. The relative intensity of fluorescence decreased immediately after the addition of ascorbic acid to the cells. The intensity from the cells treated with ascorbic acid was 20-30% of that from the control cells at each incubation time. For trolox, the intensity was 50-70% of that from the control cells at each 1 to 2 hr incubation time. When ROSs-induced lipid peroxidation was assessed using cis-parinaric acid (PnA) as a monitor molecule, lipid peroxidation was found to occur in the control cells after isolation of the bursal cells. Treatment of the cells with trolox reduced lipid peroxidation, but treatment with ascorbic acid enhanced peroxidation.

    Topics: Animals; Antioxidants; Apoptosis; Ascorbic Acid; Bursa of Fabricius; Cells, Cultured; Chickens; Chromans; DNA Fragmentation; Electrophoresis, Agar Gel; Fatty Acids, Unsaturated; Fluoresceins; Fluorescent Dyes; Lipid Peroxidation; Male; Reactive Oxygen Species; Spectrometry, Fluorescence

2000
Oxidized low density lipoprotein (ox-LDL) binding to ox-LDL receptor-1 in endothelial cells induces the activation of NF-kappaB through an increased production of intracellular reactive oxygen species.
    The Journal of biological chemistry, 2000, Apr-28, Volume: 275, Issue:17

    In this study we examined the effect of oxidized low density lipoprotein (ox-LDL) on the intracellular production of reactive oxygen species (ROS) in bovine aortic endothelial cells (BAECs) and whether this increase occurs through its binding to the endothelial receptor lectin-like ox-LDL receptor-1 (LOX-1). Furthermore, this study also aimed to ascertain whether the binding of ox-LDL to LOX-1 is associated with NF-kappaB activation. ox-LDL induced a significant dose-dependent increase in ROS production after a 30-s incubation with BAECs (p < 0.01). ROS formation was markedly reduced in BAECs incubated with anti-LOX-1 monoclonal antibody (p < 0.001), while control nonimmune IgG produced no effect. ox-LDL induced a time- and dose-dependent significant increase in ROS formation only in CHO-K1 cells stably expressing bovine LOX-1 (p < 0.001), while no increase was present in CHO-K1 cells. The activation of the transcription factor NF-kappaB in BAECs was evident after a 5-min incubation with ox-LDL and was attenuated by anti-LOX-1 monoclonal antibody. The conclusion is that one of the pathophysiological consequences of ox-LDL binding to LOX-1 may be the activation of NF-kappaB through an increased ROS production.

    Topics: Animals; Anticholesteremic Agents; Antioxidants; Ascorbic Acid; Cattle; Cells, Cultured; CHO Cells; Chromans; Cricetinae; Endothelium, Vascular; Fluoresceins; Humans; Hydrogen Peroxide; Lipoproteins, LDL; NF-kappa B; Probucol; Protein Binding; Reactive Oxygen Species; Receptors, LDL; Receptors, Oxidized LDL; Scavenger Receptors, Class E; Time Factors

2000
The use of cyclic voltammetry for the evaluation of antioxidant capacity.
    Free radical biology & medicine, 2000, Mar-15, Volume: 28, Issue:6

    Low-molecular weight antioxidants (LMWAs) play a major role in protecting biological systems against reactive oxygen-derived species and reflect the antioxidant capacity of the system. Cyclic voltammetry (CV), shown to be convenient methodology, has been validated for quantitation of the LMWA capacity of blood plasma, tissue homogenates, and plant extracts. Analysis of the CV tracing yields the values of (i) the biological oxidation potential, E and E(1/2), which relate to the nature of the specific molecule(s); (ii) the intensity (Ia) of the anodic current; and (iii) the area of the anodic wave (S). Both Ia and S relate to the concentration of the molecule(s). LMWA components of human plasma and animal tissues were identified and further validated by reconstruction of the CV tracing and by high-performance liquid chromatography-electrochemical detection. To reflect the oxidative stress status, the use of an additional parameter, R, has been proposed. R represents the level (%) of oxidized ascorbate (compared with total ascorbate) and is measured by high-performance liquid chromatography-electrochemical detection. All these parameters were monitored in healthy human subjects as well as in chronic (diabetes mellitus) and acute care patients (subjected to total body irradiation before bone marrow transplantation). The electroanalytical methodologies presented here could be widely employed for rapid evaluation of the status of subjects (in health and disease) for monitoring of their response to treatment and/or nutritional supplementation as well as for screening of specific populations.

    Topics: Amidines; Antioxidants; Ascorbic Acid; Chromans; Copper; Electrochemistry; Humans; Microelectrodes; Oxidative Stress; Peroxides; Plants, Edible

2000
Phycocyanin is an antioxidant protector of human erythrocytes against lysis by peroxyl radicals.
    The Journal of pharmacy and pharmacology, 2000, Volume: 52, Issue:4

    Topics: Antioxidants; Ascorbic Acid; Chromans; Cyanobacteria; Dose-Response Relationship, Drug; Erythrocytes; Free Radicals; Hemolysis; Humans; Peroxides; Phycocyanin

2000
Resveratrol inhibition of lipid peroxidation.
    Free radical research, 2000, Volume: 33, Issue:1

    To define the molecular mechanism(s) of resveratrol inhibition of lipid peroxidation we have utilized model systems that allow us to study the different reactions involved in this complex process. Resveratrol proved (a) to inhibit more efficiently than either Trolox or ascorbate the Fe2+ catalyzed lipid hydroperoxide-dependent peroxidation of sonicated phosphatidylcholine liposomes; (b) to be less effective than Trolox in inhibiting lipid peroxidation initiated by the water soluble AAPH peroxyl radicals; (c) when exogenously added to liposomes, to be more potent than alpha-tocopherol and Trolox, in the inhibition of peroxidation initiated by the lipid soluble AMVN peroxyl radicals; (d) when incorporated within liposomes, to be a less potent chain-breaking antioxidant than alpha-tocopherol; (e) to be a weaker antiradical than alpha-tocopherol in the reduction of the stable radical DPPH*. Resveratrol reduced Fe3+ but its reduction rate was much slower than that observed in the presence of either ascorbate or Trolox. However, at the concentration inhibiting iron catalyzed lipid peroxidation, resveratrol did not significantly reduce Fe3+, contrary to ascorbate. In their complex, our data indicate that resveratrol inhibits lipid peroxidation mainly by scavenging lipid peroxyl radicals within the membrane, like alpha-tocopherol. Although it is less effective, its capacity of spontaneously entering the lipid environment confers on it great antioxidant potential.

    Topics: Antioxidants; Ascorbic Acid; Bepridil; Biphenyl Compounds; Chromans; Free Radical Scavengers; Free Radicals; In Vitro Techniques; Iron; Lipid Peroxidation; Liposomes; Picrates; Resveratrol; Stilbenes; Vitamin E

2000
Alterations of extracellular matrix induced by tobacco smoke extract.
    Archives of dermatological research, 2000, Volume: 292, Issue:4

    Epidemiologic studies have indicated the association between tobacco smoking and skin aging, but the exact mechanism of tobacco smoke-induced premature skin aging is currently unknown. In this study, we investigated the alterations of collagen, matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in human fibroblasts treated with tobacco smoke extract. Human fibroblasts were exposed to different concentrations of water-soluble extract from tobacco smoke. Human fibroblasts irradiated with ultraviolet A1 (UVA1) were used as positive controls because the mechanism of UVA1-mediated MMP expression has been well characterized. The expression of MMP and TIMP was analyzed semiquantitatively following reverse transcriptase-polymerase chain reaction. Production of type I and type III collagens was detected by Western blotting and biosynthesis of new collagen was assessed by 3H-proline incorporation. Upon treatment with tobacco smoke extract or UVA1 irradiation, the expression of MMP-1 and MMP-3 mRNA was significantly increased in a dose-dependent manner. Maximum induction was observed with 25 microl/ml tobacco smoke extract. In contrast, the expression of TIMP-1 and TIMP-3 mRNA remained unchanged. Western blotting of the supernatant revealed that type I and type III collagens were decreased as compared with untreated controls. Collagen biosynthesis was significantly reduced by 40.1% following treatment with 25 microl/ml tobacco smoke extract. Sodium azide, L-ascorbic acid and Trolox (a water-soluble vitamin E) prevented both the UVA1- and the tobacco-induced alteration of MMP-1. These observations suggest that the imbalance of connective tissue matrix components might contribute to the molecular basis for premature skin aging in smokers. They also suggest that reactive oxygen species including singlet oxygen mediate this process.

    Topics: Adult; Aged; Antioxidants; Ascorbic Acid; Cells, Cultured; Chromans; Collagen; Collagenases; Extracellular Matrix; Female; Fibroblasts; Gene Expression Regulation; Humans; Male; Matrix Metalloproteinase 1; Matrix Metalloproteinase 3; Nicotiana; Plants, Toxic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Skin Aging; Smoke; Solutions; Tissue Inhibitor of Metalloproteinases; Ultraviolet Rays

2000
Alterations by norepinephrine of cardiac sympathetic nerve terminal function and myocardial beta-adrenergic receptor sensitivity in the ferret: normalization by antioxidant vitamins.
    Circulation, 2000, Jul-04, Volume: 102, Issue:1

    Chronic excessive norepinephrine (NE) causes cardiac sympathetic nerve terminal abnormalities, myocardial beta-adrenergic receptor downregulation, and beta-adrenergic subsensitivity. The present study was carried out to determine whether these changes could be prevented by antioxidants.. Ferrets were administered either NE (1.33 mg/d) or vehicle by use of subcutaneous pellets for 4 weeks. Animals were simultaneously assigned to receive either antioxidant vitamins (beta-carotene, ascorbic acid, and alpha-tocopherol) or placebo pellets. NE increased plasma NE 4- to 5-fold but had no effect on heart rate, heart weight, arterial pressure, or left ventricular systolic function. However, myocardial NE uptake activity and NE uptake-1 site density were reduced, as well as cardiac neuronal NE, tyrosine hydroxylase, and neuropeptide Y. In addition, there was a decrease in myocardial beta-adrenergic receptor density with a selective decrease of the beta(1)-receptor subtype, reduction of the high-affinity site for isoproterenol, decreased basal adenylyl cyclase activity, and the adenylyl cyclase responses to isoproterenol, Gpp(NH)p, and forskolin. All of these changes were prevented by antioxidant vitamins. The effects of NE on myocardial beta-adrenergic receptor density, NE uptake-1 carrier site density, and neuronal NE were also prevented by superoxide dismutase or Trolox C.. The toxic effects of NE on the sympathetic nerve terminals are mediated via the formation of NE-derived oxygen free radicals. Preservation of the neuronal NE reuptake mechanism is functionally important, because the antioxidants also prevented myocardial beta-adrenergic receptor downregulation and postreceptor abnormalities. Thus, antioxidant therapy may be beneficial in heart failure, in which cardiac NE release is increased.

    Topics: Adenylyl Cyclases; Adrenergic alpha-Agonists; Animals; Antioxidants; Ascorbic Acid; Blood Pressure; Chromans; Ferrets; Free Radical Scavengers; Heart; Heart Failure; Heart Rate; Male; Myocardium; Norepinephrine; Polyethylene Glycols; Receptors, Adrenergic, beta; Recombinant Proteins; Superoxide Dismutase; Sympathetic Nervous System; Vitamin A; Vitamin E

2000
A fluorometric method for measurement of oxygen radical-scavenging activity of water-soluble antioxidants.
    Analytical biochemistry, 2000, Aug-15, Volume: 284, Issue:1

    The relative activities of the antioxidants Trolox, ascorbic acid, uric acid, quercetin, and rutin, and the activities of total antioxidants in serum samples were determined using a fluorometric assay based on the dye 6-carboxyfluoroscein (6C-Fl) as a fluorescent indicator; 2,2'-azobis-2-amidinopropane hydrochloride (AAPH) as a peroxyl radical generator; 6-hydroxy-2,5,7, 8-tetramethyl-1-chroman-2-carboxylic acid (Trolox) as a calibrator; and phosphate buffer (pH 7.0) as a solvent. Incubation of 6C-Fl in 0. 075 M phosphate buffer, in the presence of AAPH at 37 degrees C, resulted in loss of its fluorescence signal at 520 nm with excitation at 495 nm. The antioxidants Trolox, ascorbic acid, and uric acid provided protection of the fluorescence of 6C-Fl, and the relative antioxidant activities, determined by the net protection area under curve technique, were found to be 1:0.4:1, respectively. Trolox and ascorbic acid were used to validate this assay. A linear correlation of the net protection value with the concentration of serum, Trolox, ascorbic acid, and uric acid was demonstrated. Quercetin and rutin were shown to have strong antioxidant activities, nearly 10 times those of vitamin C. This assay is simple, reliable, and suitable for automation to handle many samples and requires few microliters of serum samples.

    Topics: Amidines; Animals; Antioxidants; Area Under Curve; Ascorbic Acid; Chromans; Dogs; Fluoresceins; Free Radicals; Humans; Oxygen; Quercetin; Rutin; Spectrometry, Fluorescence; Time Factors; Uric Acid

2000
Melatonin but not vitamins C and E maintains glutathione homeostasis in t-butyl hydroperoxide-induced mitochondrial oxidative stress.
    FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 2000, Volume: 14, Issue:12

    Topics: Animals; Antioxidants; Ascorbic Acid; Chromans; Glutathione; Glutathione Disulfide; Homeostasis; Melatonin; Mitochondria; Oxidation-Reduction; Oxidative Stress; Rats; tert-Butylhydroperoxide; Vitamin E

2000
Comparative effects of melatonin, L-deprenyl, Trolox and ascorbate in the suppression of hydroxyl radical formation during dopamine autoxidation in vitro.
    Journal of pineal research, 2000, Volume: 29, Issue:2

    Degeneration of nigrostriatal dopaminergic neurons is the major pathogenic substrate of Parkinson's disease (PD). Inhibitors of monoamine oxidase B (MAO-B) have been used in the treatment of PD and at least one of them, i.e., deprenyl, also displays antioxidant activity. Dopamine (DA) autoxidation produces reactive oxygen species implicated in the loss of dopaminergic neurons in the nigrostriatal pathway. In this study we compared the effects of melatonin with those of deprenyl and vitamins E and C in preventing the hydroxyl radical (8OH) generation during DA oxidation. The rate of production of 2,3-dihydroxybenzoate (2,3-DHBA) in the presence of salicylate, an *OH scavenger, was used to detect the in vitro generation of *OH during iron-catalyzed oxidation of DA. The results showed a dose-dependent effect of melatonin, deprenyl and vitamin E in counteracting DA autoxidation, whereas vitamin C had no effect. Comparative analyses between the effect of these antioxidants showed that the protective effect of melatonin against DA autoxidation was significantly higher than that of the other compounds tested. Also, when melatonin plus deprenyl were added to the incubation medium, a potentiation of the antioxidant effect was found. These findings suggest that antioxidants may be useful in brain protection against toxicity of reactive oxygen species produced during DA oxidation, and melatonin, alone or in combination with deprenyl, may be an important component of the brain's antioxidant defenses to protect it from dopaminergic neurodegeneration.

    Topics: Antioxidants; Ascorbic Acid; Chromans; Dopamine; Dose-Response Relationship, Drug; Hydroxybenzoates; Hydroxyl Radical; Iron; Melatonin; Oxidation-Reduction; Salicylates; Selegiline

2000
Ascorbate-dependent recycling of the vitamin E homologue Trolox by dihydrolipoate and glutathione in murine skin homogenates.
    Free radical biology & medicine, 2000, Volume: 29, Issue:3-4

    In the redox antioxidant network, dihydrolipoate can synergistically enhance the ascorbate-dependent recycling of vitamin E. Since the major endogenous thiol antioxidant in biological systems is glutathione (GSH) it was of interest to compare the effects of dihydrolipoate with GSH on ascorbate-dependent recycling of the water-soluble homologue of vitamin E, Trolox, by electron spin resonance (ESR). Trolox phenoxyl radicals were generated by a horseradish peroxidase (HRP)-hydrogen peroxide (H2O2) oxidation system. In the presence of dihydrolipoate, Trolox radicals were suppressed until both dihydrolipoate and endogenous levels of ascorbate in skin homogenates were consumed. Similar experiments made in the presence of GSH revealed that Trolox radicals reappeared immediately after ascorbate was depleted and that GSH was not able to drive the ascorbate-dependent Trolox recycling reaction. However, at higher concentrations GSH was able to increase ascorbate-mediated Trolox regeneration from the Trolox radical. ESR and spectrophotometric measurements demonstrated the ability of dihydrolipoate or GSH to react with dehydroascorbate, the two-electron oxidation product of ascorbate in this system. Dihydrolipoate regenerated greater amounts of ascorbate at a much faster rate than equivalent concentrations of GSH. Thus the marked difference between the rate and efficiency of ascorbate generation by dihydrolipoate as compared with GSH appears to account for the different kinetics by which these thiol antioxidants influence ascorbate-dependent Trolox recycling.

    Topics: Animals; Antioxidants; Ascorbic Acid; Cell Extracts; Chromans; Electron Spin Resonance Spectroscopy; Free Radicals; Glutathione; Hydrogen Peroxide; Kinetics; Mice; Peroxidase; Phenols; Skin; Thioctic Acid; Vitamin E

2000
Novel approach to in vivo screening for radioprotective activity in whole mice: in vivo electron spin resonance study probing the redox reaction of nitroxyl.
    Journal of radiation research, 2000, Volume: 41, Issue:2

    Previously, we reported that X-irradiation enhanced the signal decay of a spin probe injected into whole mice measured by in vivo ESR, and that the observed enhancement was suppressed by the pre-administration of cysteamine, a radioprotector [Miura, Y., Anzai, K., Urano, S. and Ozawa, T. (1997) Free Rad. Biol. Med. 23: 533-540]. In the present study, the suppression activity of the X-ray-induced increase in the ESR signal decay rate (termed suppression index, SI) was measured for several radioprotectors: 5-hydroxytryptamine (5-HT), S-2-(3-aminopropylamino)-ethylphosphorothioic acid (WR-2721), 4-hydroxy-2,2,6,6-tetramethyl-piperidine-N-oxyl (TEMPOL), cimetidine, interleukin-1 beta (IL-1 beta) and stem cell factor (SCF). The enhancement of the ESR signal decay of carbamoyl-PROXYL due to X-irradiation was suppressed by a treatment with all of the radioprotectors examined, showing positive SI values. However, a dose-dependency of 5-HT or WR-2721 was not observed, suggesting that several mechanisms exist for radioprotection and a modification of the signal decay rate. Although the in vivo ESR system cannot be used in place of the 30-day survival method for the assessment of radioprotectors, this system might be applicable to in vivo, non-invasive screening prior to using the 30-day survival method.

    Topics: Amifostine; Animals; Ascorbic Acid; Chromans; Cimetidine; Cyclic N-Oxides; Dose-Response Relationship, Drug; Electron Spin Resonance Spectroscopy; Interleukin-1; Mice; Nitrogen Oxides; Oxidation-Reduction; Oxidative Stress; Pyrrolidines; Radiation-Protective Agents; Recombinant Proteins; Serotonin; Spin Labels; Stem Cell Factor; Vitamin E; Whole-Body Irradiation

2000
Protective effect of beta-carbolines and other antioxidants on lipid peroxidation due to hydrogen peroxide in rat brain homogenates.
    Neuroscience letters, 2000, Nov-10, Volume: 294, Issue:1

    Tryptoline and pinoline are two beta-carbolines isolated from the nervous system of mammals. We investigated the ability of these compounds to prevent lipid peroxidation induced by hydrogen peroxide in rat brain homogenates. We also compared their effects with other known antioxidants including melatonin, trolox and ascorbic acid. Lipid peroxidation was assessed by measuring malonaldehyde (MDA) and 4-hydroxy-alkenals (4-HDA) concentrations in the brain homogenates. Incubation with hydrogen peroxide (5 mM) increased MDA+4-HDA levels, which were totally prevented by tryptoline, pinoline, melatonin and trolox in a concentration-dependent manner. By contrast, higher MDA-4-HDA concentrations compared with control experiments were found after incubation with ascorbic acid, thus reflecting an increase of lipid peroxidation induced by this compound. Although in vivo studies are needed, the data suggest that these beta-carbolines may be potential neuroprotective agents because of their antioxidant activities.

    Topics: Aldehydes; Animals; Antioxidants; Ascorbic Acid; Brain Chemistry; Carbolines; Cell-Free System; Chromans; Dose-Response Relationship, Drug; Hydrogen Peroxide; Lipid Peroxidation; Male; Malondialdehyde; Melatonin; Rats; Rats, Sprague-Dawley

2000
Diet enriched with procyanidins enhances antioxidant activity and reduces myocardial post-ischaemic damage in rats.
    Life sciences, 1999, Volume: 64, Issue:8

    Aim of this work was to study the efficacy of procyanidins from Vitis vinifera seeds, a standardized mixture of polyphenol antioxidants, on cardiac mechanics following ischemia/reperfusion stunning in the rat, after 3 weeks supplementation. Young and aged male rats were fed a diet enriched with procyanidins complexed (1:3 w/w) with soybean lecithin (2.4%); control animals (CTR-young and CTR-aged) received an equal amount of lecithin and 2 additional groups of animals the standard diet. At the end of the treatment, the total plasma antioxidant defense (TRAP), vitamin E, ascorbic acid and uric acid were determined in plasma and the hearts from all groups of animals subjected to moderate ischemia (flow reduction to 1 ml/min for 20 min) and reperfusion (15 ml/min for 30 min). In both young and aged rats supplemented with procyanidins the recovery of left ventricular developed pressure (LVDP) at the end of reperfusion was 93% (p < 0.01) and 74% (p < 0.01) of the preischemic values and the values of coronary perfusion pressure (CPP) were maintained close to those of the preischemic period. Also creatine kinase (CK) outflow was restrained to baseline levels, while a 2-fold increase in prostacyclin (6-keto-PGF1alpha) in the perfusate from hearts of young and aged rats was elicited during both ischemia and reperfusion. In parallel, procyanidins significantly increased the total antioxidant plasma capacity (by 40% in young and by 30% in aged rats) and the plasma levels of ascorbic acid, while tend to reduce vitamin E levels; no significant differences were observed in uric acid levels. The results of this study demonstrate that procyanidins supplementation in the rat (young and aged) makes the heart less susceptible to ischemia/reperfusion damage and that this is positively associated to an increase in plasma antioxidant activity.

    Topics: Aging; Angiotensin II; Animals; Antioxidants; Ascorbic Acid; Biflavonoids; Blood Glucose; Body Weight; Catechin; Cholesterol; Chromans; Creatine Kinase; Dietary Supplements; Epoprostenol; Heart; Male; Myocardial Reperfusion Injury; Myocardium; Proanthocyanidins; Rats; Rats, Wistar; Triglycerides; Uric Acid; Vitamin E

1999
Effective inhibition by beta-carotene of cellular DNA breaking induced by peroxynitrous acid.
    Free radical research, 1999, Volume: 30, Issue:1

    Peroxynitrous acid synthesized by reaction of hydrogen peroxide and nitrite and generated from 3-morpholinosydononimine (SIN-1) induced cellular DNA breaking of human promyelocytic leukemia HL-60 cells in phosphate buffer (pH 7.5) as assessed by alkaline single cell gel electrophoresis (comet) assay and quantification of comet types. Ascorbate and Trolox inhibited cellular DNA breaking induced by peroxynitrous acid, but the concentrations of these antioxidants required for effective inhibition was about 50-fold higher than that of peroxynitrous acid. beta-Carotene protected DNA breaking by peroxynitrous acid in 20% tetrahydrofuran-phosphate buffer (pH 7.5) much more effectively than ascorbate and Trolox. The concentrations of beta-carotene required for effective inhibition was lower than the concentration of peroxynitrous acid.

    Topics: Antioxidants; Ascorbic Acid; beta Carotene; Chromans; Dimethyl Sulfoxide; DNA; DNA Damage; Dose-Response Relationship, Drug; Electrophoresis, Agar Gel; Free Radical Scavengers; Furans; HL-60 Cells; Humans; Mannitol; Microscopy, Fluorescence; Molsidomine; Nitrous Acid; Peroxynitrous Acid; Solubility; Sorbic Acid; Time Factors

1999
Antioxidants reversibly inhibit the spontaneous resumption of meiosis.
    The American journal of physiology, 1999, Volume: 276, Issue:4

    We previously showed that the cell-permeant antioxidant 2(3)-tert-butyl-4-hydroxyanisole (BHA) inhibited germinal vesicle breakdown (GVBD) in oocyte-cumulus complexes (OCC) of the rat. The objective of the present studies was to assess other antioxidants and whether such inhibition was reversible. Spontaneous GVBD in OCC incubated for 2 h was significantly inhibited (P < 0.005) by nordihydroguaiaretic acid (NDGA; GVBD = 19.4%), BHA (GVBD = 25.7%), octyl gallate (OG; GVBD = 52.2%), ethoxyquin (EQ; GVBD = 58.8%), 2, 6-di-tert-butyl-hydroxymethyl phenol (TBHMP; GVBD = 59%), butylated hydroxytoluene (BHT; GVBD = 59.5%), and tert-butyl hydroperoxide (TBHP; GVBD = 60.0%). Other antioxidants that produced lower but significant (P < 0.05) inhibition of oocyte maturation included propyl gallate (PG; GVBD = 70.3%), 2,4,5-trihydroxybutrophenone (THBP; GVBD = 71.4%), and lauryl gallate (LG; GVBD = 71.4%). Antioxidants that had no effect on oocyte maturation at the same concentration (100 microM) included ascorbic acid, vitamin E, and Trolox. Inhibition of GVBD was evident for up to 8 h of incubation of OCC and denuded oocytes (DO) with BHA or NDGA and was reversed by washing. NDGA was less potent than BHA for inhibition of GVBD in DO, unlike that seen with OCC. Oocyte maturation was induced by incubation of follicles for 3 h with human chorionic gonadotropin (hCG), and this response was inhibited by BHA or NDGA. These findings support the conclusion that cell-permeant antioxidants inhibit spontaneous resumption of meiosis, which may implicate a role of oxygen radicals in oocyte maturation.

    Topics: Animals; Antioxidants; Ascorbic Acid; Butylated Hydroxytoluene; Chorionic Gonadotropin; Chromans; Female; Humans; Hydroquinones; Masoprocol; Meiosis; Oocytes; Ovarian Follicle; Rats; Rats, Sprague-Dawley; Vitamin E

1999
Metalloporphyrins are potent inhibitors of lipid peroxidation.
    Free radical biology & medicine, 1999, Volume: 26, Issue:5-6

    The objectives of these studies were to determine whether metalloporphyrins could inhibit lipid peroxidation, characterize factors that influence their potency and compare their potency to prototypical antioxidants. Lipid peroxidation was initiated with iron and ascorbate in rat brain homogenates and the formation of thiobarbituric acid reactive species was used as an index of lipid peroxidation. Metalloporphyrins were found to be a novel and potent class of lipid peroxidation inhibitors. Inhibition of lipid peroxidation by metalloporphyrins was dependent on the transition metal ligated to the porphyrin, indicating that metal centered redox chemistry was important to the mechanism of their antioxidant activities. Manganese porphyrins with the highest superoxide dismutase (SOD) activities, MnOBTM-4-PyP and MnTM-2-PyP (charges are omitted throughout text for clarity), were the most potent inhibitors of lipid peroxidation with calculated IC50s of 1.3 and 1.0 microM, respectively. These manganese porphyrins were 2 orders of magnitude more potent than either trolox (IC50 = 204 microM) or rutin (IC50 = 112 microM). The potencies of the manganese porphyrins were related not only to their redox potentials and SOD activities, but also to other factors that may contribute to their ability to act as electron acceptors. The broad array of antioxidant activities possessed by metalloporphyrins make them attractive therapeutic agents in disease states that involve the overproduction of reactive oxygen species.

    Topics: Animals; Antioxidants; Ascorbic Acid; Brain; Chromans; Free Radical Scavengers; Kinetics; Lipid Peroxidation; Manganese; Metalloporphyrins; Oxidation-Reduction; Rats; Rats, Sprague-Dawley; Rutin; Superoxide Dismutase; Zinc

1999
Antioxidant properties of EPC-K1: a study on mechanisms.
    Biophysical chemistry, 1999, Mar-29, Volume: 77, Issue:2-3

    Scavenging effects of L-ascorbic acid 2-[3,4-dihydro-2,5,7,8- tetramethyl-2-(4,8,12-trimethytridecyl)-2H-1-benzopyran- 6-yl-hydrogen phosphate] potassium salt (EPC-K1) on hydroxyl radicals, alkyl radicals and lipid radicals were studied with ESR spin trapping techniques. The inhibition effects of EPC-K1 on lipid peroxidation were assessed by TBA assay. The kinetics of EPC-K1 reacting with hydroxyl radicals and linoleic acid radicals were studied by pulse radiolysis. The active site of EPC-K1 and the structure-antioxidative activity relationships were discussed. The superoxide radicals scavenging capacity of the brain homogenate of EPC-K1-treated rats was measured. The results revealed that in comparison with Trolox and vitamin C, EPC-K1 showed better overall antioxidative capacity in vitro and in vivo. EPC-K1 was a moderate scavenger on hydroxyl radicals and alkyl radicals, a potent scavenger on lipid radicals, and an effective inhibitor on lipid peroxidation. EPC-K1 could react with hydroxyl radicals with a rate constant of 7.1 x 10(8) dm3 mol-1 s-1 and react with linoleic acid radicals with a rate constant of 2.8 x 10(6) dm3 mol-1 s-1. The active site of EPC-K1 was the enolic hydroxyl group. After administration of EPC-K1, the ability of rat brain to scavenge superoxide radicals was significantly increased. The potent scavenging effects of EPC-K1 on both hydrophilic and hydrophobic radicals were relevant with its molecular structure, which consisted of both hydrophilic and hydrophobic groups.

    Topics: Animals; Antioxidants; Ascorbic Acid; Brain; Chromans; Electron Spin Resonance Spectroscopy; Free Radical Scavengers; Hydroxyl Radical; Linoleic Acid; Lipid Peroxidation; Male; Molecular Structure; Peroxides; Pulse Radiolysis; Rats; Rats, Wistar; Spectrophotometry; Structure-Activity Relationship; Vitamin E

1999
The importance of lipid solubility in antioxidants and free radical generating systems for determining lipoprotein proxidation.
    Free radical biology & medicine, 1999, Volume: 26, Issue:11-12

    The oxidative modification of low-density lipoprotein (LDL) plays an important role in atherosclerosis. Protecting LDL from oxidation has been shown to reduce the risk of coronary heart disease. In this study, we compared the protective effects of two lipophilic antioxidants (vitamin E and lazaroid) with two hydrophilic antioxidants (trolox and vitamin C) in the presence of several different free radical generating systems. Vitamin E (IC50 = 5.9 microM) and lazaroid (IC50 = 5.0 microM) were more effective in inhibiting lipid peroxidation caused by a Fe-ADP free radical generating system than vitamin C (IC50 = 5.2 x 10(3) microM) and trolox (IC5 = 1.2 x 10(3) microM). Preincubation of lipoproteins with a lipophilic antioxidant increased the protective effect against various free radicals. Preincubation with hydrophilic antioxidants did not have an effect. We also tested the efficacy of the antioxidants when the free radicals were generated within the lipid or the aqueous environment surrounding the LDL. For this purpose, we used the peroxyl generating azo-compounds AMVN (2,2'-azobis(2,4-dimethylvaleronitrile)) and AAPH (2,2'azobis(2-amidinopropane) dihydrochloride). All of the antioxidants tested were more effective against free radicals generated in a water soluble medium than they were against free radicals generated in a lipid environment. In conclusion, our data demonstrate that lipid solubility is an important factor for both the antioxidant and the free radical generating systems in determining the extent of lipid peroxidation in LDL. Our data also demonstrate that antioxidant efficacy in one set of experimental conditions may not necessarily translate into a similar degree of protection in another set of conditions where lipophilicity is a variable.

    Topics: Animals; Antioxidants; Ascorbic Acid; Chromans; Free Radicals; Lipid Peroxidation; Lipids; Lipoproteins; Male; Rabbits; Solubility; Vitamin E; Water

1999
ESR studies of ascorbic acid-dependent recycling of the vitamin E homologue Trolox by coenzyme Q0 in murine skin homogenates.
    Redox report : communications in free radical research, 1999, Volume: 4, Issue:3

    The recycling of Trolox, a water-soluble vitamin E homologue, by coenzyme Q0 (CoQ0) during Cu2+-initiated oxidation of ascorbate in mouse skin homogenates was investigated using electron spin resonance (ESR) spectroscopy. In a mixture containing CoQ0, Cu2+ and mouse skin homogenates, the ESR signal of CoQ0 semiquinone radical (CoQ0*-) appeared and declined with time; addition of Trolox accelerated the CoQ0*- signal decay. Only after the disappearance of the CoQ0*- signal was the appearance of the Trolox phenoxyl radical signal observed. In addition, the lifetime of the CoQ0*- signal and the length of the lag period during which the Trolox radical ESR signal could not be detected were dependent on the presence of Trolox, CoQ0 or Cu2+. The results suggest that CoQ0*-, formed by the interaction between CoQ0 and endogenous ascorbic acid (AscH-) in skin homogenates, regenerates Trolox from its phenoxyl radical.

    Topics: Animals; Antioxidants; Ascorbic Acid; Chromans; Coenzymes; Copper; Electron Spin Resonance Spectroscopy; Female; Free Radicals; Kinetics; Mice; Mice, Hairless; Models, Chemical; Skin; Ubiquinone

1999
Antioxidant transport modulates peripheral airway reactivity and inflammation during ozone exposure.
    Journal of applied physiology (Bethesda, Md. : 1985), 1999, Volume: 87, Issue:5

    We examined the effects of ozone (O(3)) and endogenous antioxidant transport on canine peripheral airway function, central airway function, epithelial integrity, and inflammation. Dogs were either untreated or pretreated with probenecid (an anion-transport inhibitor) and exposed for 6 h to 0.2 parts/million O(3). Peripheral airway resistance (Rpa) and reactivity (DeltaRpa) were monitored in three sublobar locations before and after exposure to either air or O(3). Pulmonary resistance and transepithelial potential difference in trachea and bronchus were also recorded. Bronchoalveolar lavage fluid (BALF) was collected before, during, and after exposure. O(3) increased Rpa and DeltaRpa only in probenecid-treated dogs and in a location-dependent fashion. Pulmonary resistance and potential difference in bronchus increased after O(3) exposure regardless of treatment. O(3) markedly increased BALF neutrophils only in untreated dogs. With the exception of hexanal, O(3) did not alter any BALF constituent examined. Probenecid reduced BALF ascorbate, BALF protein, and plasma urate. We conclude that 1) a 6-h exposure to 0.2 parts/million O(3) represents a subthreshold stimulus in relation to its effects on peripheral airway function in dogs, 2) antioxidant transport contributes to the maintenance of normal airway tone and reactivity under conditions of oxidant stress, 3) O(3)-induced changes in Rpa and DeltaRpa are dependent on location, and 4) peripheral airway hyperreactivity and inflammation reflect independent responses to O(3) exposure. Finally, although anion transport mitigates the effect of O(3) on peripheral airway function, it contributes to the development of airway inflammation and may represent a possible target for anti-inflammatory prevention or therapy.

    Topics: Airway Resistance; Aldehydes; Animals; Antioxidants; Ascorbic Acid; Biological Transport, Active; Bronchoalveolar Lavage Fluid; Cell Count; Chromans; Dogs; Inflammation; Male; Oxidants, Photochemical; Ozone; Peroxides; Probenecid; Proteins; Respiratory System; Uricosuric Agents

1999
Effect of antioxidant supplementation on the adaptive response of human skin fibroblasts to UV-induced oxidative stress.
    Redox report : communications in free radical research, 1999, Volume: 4, Issue:6

    The effect of supplementation with substances having antioxidant properties on the adaptive responses of human skin fibroblasts to UV-induced oxidative stress was studied in vitro. UVR was found to induce a substantial oxidative stress in fibroblasts, resulting in an increased release of superoxide anions and an increase in lipid peroxidation (shown by an elevated malonaldehyde content). Sub-lethal doses of UVR were also found to induce adaptive responses in the fibroblast antioxidant defences, with a transient rise in catalase and superoxide dismutase activities followed by a slower, large increase in cellular glutathione content. Supplementation of the fibroblasts with the antioxidants, Trolox (a water soluble analogue of alpha-tocopherol), ascorbic acid or beta-carotene, had differential effects on these responses. Trolox supplementation reduced the UVR-induced cellular oxidative stress and adaptive response in a predictable concentration-dependent manner. This was in contrast to ascorbic acid which increased superoxide release from fibroblasts. At low doses, ascorbate supplements also reduced the magnitude of the adaptive increases in catalase and superoxide dismutase activities and increase in glutathione content. Beta-carotene had a similar effect to ascorbic acid, reducing the extent of the adaptations to UVR at lower doses while simultaneously increasing superoxide release and malonaldehyde content. These in vitro data indicate that only the vitamin E analogue suppressed UVR-induced oxidative stress in a predictable manner and suggest that common dietary antioxidants may not be equally effective in reducing the potential deleterious effects of UVR-induced oxidative stress in skin.

    Topics: Adaptation, Physiological; Antioxidants; Ascorbic Acid; beta Carotene; Cell Line; Chromans; Dose-Response Relationship, Radiation; Fibroblasts; Humans; Oxidative Stress; Skin; Superoxides; Ultraviolet Rays

1999
A rapid gas chromatographic assay for determining oxyradical scavenging capacity of antioxidants and biological fluids.
    Free radical biology & medicine, 1998, Volume: 24, Issue:3

    Herein, we report a new, rapid,and reliable method for measuring the protective antioxidant potential of pure antioxidant solutions or biological tissues. Peroxyl radicals generated by thermal homolysis of 2,2'-azobis-amidinopropane (ABAP) cause the oxidation of alpha-keto-gamma-methiolbutyric acid (KMBA) to ethylene; ethylene formation is monitored by gas chromatographic analysis of head space from the reaction vessel. The partial inhibition of ethylene formation in the presence of antioxidants that compete with KMBA for oxyradicals is the basis of the Total Oxyradical Scavenging Capacity Assay (TOSCA). The assay is shown to be reliable for quantifying ROS scavenging potential. The quantifiable parameters are consistent with the relative order of those predicted by the fluorescence- and oxygen electrode-based assays reported in the literature. Antioxidants competing for peroxyl radicals influenced the rate of KMBA oxidation in different ways, but the calculation of TOSC was not affected by such variations. Responses were linear over a wide range of sample concentrations and the TOSC values of classical soluble antioxidants showed the following relative order: Trolox > uric acid > ascorbic acid > GSH. The KMBA method was reliable for biological tissues; the TOSC for 1 microg rat liver cytosolic protein was 0.40 +/- 0.02 and for the microsomal membrane, 0.15 +/- 0.03. Soluble antioxidants accounted for 77% of the protective antioxidant potential in rat liver cytosol. When incorporated into the microsomal membrane, alpha-tocopherol markedly enhances antioxidant protection against peroxyl radical; thus, the assay is suitable for the assessment of fat-soluble antioxidants.

    Topics: Animals; Antioxidants; Ascorbic Acid; Body Fluids; Butyrates; Chromans; Chromatography, Gas; Cytosol; Ethylenes; Free Radical Scavengers; Glutathione; Kinetics; Liver; Microsomes; Peroxides; Rats; Solutions; Sulfhydryl Compounds; Uric Acid; Vitamin E

1998
Sustained hypoxia-ischemia results in reactive nitrogen and oxygen species production and injury in the premature fetal rabbit brain.
    Journal of neuropathology and experimental neurology, 1998, Volume: 57, Issue:6

    Free radical-mediated injury is implicated in hypoxic-ischemic encephalopathy observed in neonates. We investigated in utero free radical production and injury following hypoxia-ischemia to premature fetal brain utilizing a rabbit model of acute placental insufficiency. Pregnant rabbits at 29 days gestation were randomized to uterine ischemia for 50 minutes (min) (hypoxia) or nonischemic controls. Fetal brains were obtained immediately after ischemia for oxidative and acute-injury markers or 24 hours (h) post-ischemia for histopathology. Nitrotyrosine formation, a marker of NO-derived species such as peroxynitrite, was observed only in hypoxic brains. Hypoxia resulted in a significant increase in nitrogen oxides, lipid peroxidation, and protein oxidation, with a concomitant decrease in total antioxidant capacity, compared with controls. Peroxynitrite addition to brain homogenate increased nitrogen oxides linearly (1:1), although protein carbonyls were unchanged. Concomitantly, in vitro cortical and hippocampal cell viability and ATP levels decreased, with an increase in brain edema in hypoxic brains. Fetuses delivered 24 h post-ischemia had increased hippocampal nuclear karyorrhexis on histology compared with controls. Antioxidant administration (ascorbic acid and Trolox) intraperitoneally ameliorated changes in cellular viability and brain edema. Acute fetal hypoxia-ischemia without reoxygenation results in increased nitrogen and oxygen free radical production that may cause brain injury. The merits of the described model are discussed.

    Topics: Animals; Antioxidants; Ascorbic Acid; Brain; Brain Ischemia; Cell Death; Cell Survival; Chromans; Female; Free Radicals; Hypoxia, Brain; Nitrogen; Oxidative Stress; Pregnancy; Rabbits; Reactive Oxygen Species; Uterus

1998
Possible relationship between changes in [3H]DA uptake and autoxidation in rat striatal slices.
    Experimental neurology, 1998, Volume: 152, Issue:1

    Many recent studies have suggested that oxidative damage is an important factor in several neurodegenerative disorders. Our investigations considered whether autoxidation of rat striatal slices modified dopamine uptake. Biochemical assays (TBARS, MDA-TBA complex, aldehydes, and fluorescent lipid-soluble products) and a [3H]DA uptake assay were performed on nonincubated striatal slices and on slices incubated for 150 min at 37 degreesC in Krebs-Ringer buffer without addition of free-radical generators. The results showed that spontaneous lipid peroxidation occured during incubation and that DA uptake kinetic was biphasic (high-affinity uptake1 and low-affinity uptake2) with a significant decrease of maximal velocity of uptake. Ascorbate, a known antioxidant, was used to determine whether a relationship existed between lipid peroxidation and reduced dopamine uptake. Addition of ascorbate (100 and 500 microM) in Krebs-Ringer buffer for 150 min at 37 degreesC failed to indicate whether decreased [3H]DA uptake resulted from lipid peroxidation. In fact, ascorbate acted as a prooxidant, only preventing decreased DA uptake2 at 100 microM. Trolox, another antioxidant, inhibited lipid peroxidation by about 95% with a concentration of 700 microM and protected only uptake1. With a concentration of 5000 microM, Trolox also protected uptake2. On the whole, these results indicate that spontaneous autoxidation in rat striatal slices was associated with a lipid peroxidation process that altered the DA uptake system.

    Topics: Animals; Antioxidants; Ascorbic Acid; Carrier Proteins; Chromans; Dopamine; Dopamine Plasma Membrane Transport Proteins; Fluorescent Dyes; In Vitro Techniques; Lipid Peroxidation; Male; Malondialdehyde; Membrane Glycoproteins; Membrane Transport Proteins; Neostriatum; Nerve Tissue Proteins; Oxidation-Reduction; Presynaptic Terminals; Rats; Rats, Wistar; Thiobarbituric Acid Reactive Substances

1998
Simple determination of peroxyl radical-trapping capacity.
    Biochemistry and molecular biology international, 1998, Volume: 46, Issue:3

    A simple spectrophotometric method of determination of peroxyl radical-trapping capacity (PRTC) of body fluids and food products is proposed. In this method, decomposition of 2,2'-azobis(2-amidopropane) hydrochloride (ABAP) is the source of peroxyl and alkoxyl radicals which oxidize 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) to a green cation radical. Antioxidant present in a sample inhibit the reaction; the induction time of the reaction is proposed as a parameter enabling determination of antioxidant content. Standard assay conditions are: 20 mM ABAP and 150 microM ABTS in 0.1 M phosphate buffer, pH 7.0, at 37 degrees C; absorbance is monitored at 414 nm. A 10-min assay allows for determination of the induction time of appropriately diluted sample. As examples of application of this method, PRTC values of several types of beverages are reported.

    Topics: Alcoholic Beverages; Amidines; Amino Acids; Antioxidants; Ascorbic Acid; Benzothiazoles; Chromans; Epinephrine; Free Radicals; Indicators and Reagents; Oxidation-Reduction; Peroxides; Plasma; Spectrophotometry; Sulfonic Acids; Time Factors

1998
Inhibitory effects of ascorbic acid on dopamine uptake by rat striatal synaptosomes: relationship to lipid peroxidation and oxidation of protein sulfhydryl groups.
    Neuroscience research, 1998, Volume: 32, Issue:2

    Ascorbic acid is frequently added in the incubation medium to prevent oxidation of dopamine (DA) during uptake assays. However, a preliminary study showed that the presence of ascorbic acid induced a decrease of DA uptake after prolonged incubation. The purpose of this study was to determine the mechanism underlying ascorbic acid-induced alterations of DA uptake in rat striatal synaptosomes. In this context, the effects of physiological concentrations of ascorbic acid (100-500 microM) on DA uptake and Na+/K+ ATPase activity (which is essential for DA transporter function) were assessed in synaptosomes before and after incubation at 37 degrees C. The capacity of synaptosomes to take up DA was significantly decreased after incubation owing to a reduction in DA transporters (but with no modification of their affinity for DA). This partial inhibition was associated with a decrease of Na+/K+ ATPase activity, a production of thiobarbituric acid reactive substances (TBARS) and malonaldehyde (MDA), and a loss of sulfhydryl group content. Addition of Trolox C to the medium prevented the reduction of DA uptake, the inhibition of Na+/K+ ATPase activity, the decrease in sulfhydryl group content and the production of TBARS and MDA. These results suggest that ascorbic acid in the presence of contaminant ferrous ions induced a decrease in functional DA transporters, probably through a lipid peroxidation process involving oxidation of sulfhydryl groups and at least in part through a decrease of Na+/K+ ATPase activity.

    Topics: Animals; Ascorbic Acid; Biological Transport; Chromans; Corpus Striatum; Dopamine; Lipid Peroxidation; Malondialdehyde; Oxidation-Reduction; Proteins; Rats; Rats, Wistar; Sodium-Potassium-Exchanging ATPase; Sulfhydryl Compounds; Synaptosomes; Thiobarbituric Acid Reactive Substances; Time Factors

1998
Effects of vitamins E, C and catalase on bromobenzene- and hydrogen peroxide-induced intracellular oxidation and DNA single-strand breakage in Hep G2 cells.
    Journal of hepatology, 1997, Volume: 26, Issue:3

    Water-soluble vitamin E (Trolox C), ascorbic acid and catalase were shown in our previous study to protect isolated rat hepatocytes against bromobenzene-induced toxicity.. In order to study the mechanisms of this protection and the pathogenesis of bromobenzene-induced hepatocellular injury, a fluorometric assay for the investigation of intracellular oxidation, indicated by conversion of dichlorofluorescein diacetate to dichlorofluorescein, was used. Single-strand DNA breakage was also evaluated in Hep G2 cells by a radio-labelling method.. Bromobenzene (2.4 and 4.8 mM) induced a significant increase in dichlorofluorescein fluorescence intensity compared to the controls. Trolox C, ascorbic acid or catalase significantly inhibited bromobenzene-induced enhancement of fluorescence intensity (p<0.05-0.001), as well as reduced auto-intracellular oxidation in untreated Hep G2 cells. Hydrogen peroxide (H2O2) evoked a dose-dependent increase in dichlorofluorescein fluorescence intensity in Hep G2 cells, and the effect was completely blocked by Trolox C (2.0 mM) and catalase (4800 unit/ml). Bromobenzene caused significant single-strand DNA breakage in Hep G2 cells during 2 h suspension incubation and 24 h primary incubation. H2O2 (400 microM) led to marked single-strand DNA breakage in 20 min, and the effect was attenuated by Trolox C.. Metabolism of bromobenzene in Hep G2 cells induces production of H2O2, indicated by enhancement of dichlorofluorescein fluorescence intensity, or other free radicals, which leads to single-strand DNA breakage in the cells. Vitamins E and C and catalase display strong intracellular antioxidative effects. Vitamin E could partially inhibit H2O2-induced single-strand DNA breakage in the cells.

    Topics: Antioxidants; Ascorbic Acid; Bromobenzenes; Catalase; Cells, Cultured; Chromans; DNA Damage; DNA, Single-Stranded; Dose-Response Relationship, Drug; Fluoresceins; Fluorescence; Humans; Hydrogen Peroxide; Intracellular Fluid; Liver; Oxidants; Oxidation-Reduction; Vitamin E

1997
Oxidative tissue response promoted by 5-aminolevulinic acid promptly induces the increase of plasma antioxidant capacity.
    Free radical research, 1997, Volume: 26, Issue:3

    The heme precursor 5-aminolevulinic acid (ALA), acting as a prooxidant, has been proposed to underlie the clinical manifestations of various porphyric disorders. Accordingly, ALA-generated oxyradicals where shown to cause oxidative lesions in biomolecules and isolated cell organelles and to release iron from ferritin. In rats, administered ALA triggered oxidative stress in liver, brain and red muscles. We now study the correlation between the plasma antioxidant capacity and tissue oxidative damage, after acute (one and two doses) and prolonged (eight doses) ALA treatment of rats (one dose of ALA = 40 mg/kg body weight). The in situ spontaneous chemiluminescence intensity increased 5-fold in brain, 50% in liver and 4-fold in soleus muscle upon two dose-treatment, indicating tissue response to oxidative injury by ALA. Chemiluminescence reached the highest intensity after one or two doses of ALA and decreased after eight doses in all tissues. The plasma trapping capacity, evaluated by the luminol/2-amidinopropane system, gave a parallel response: maximum values after two doses and decreased values after prolonged treatment. After eight doses, the ALA concentration was found to be 3-fold above the normal value in plasma, 48% higher in liver and 38% higher in total brain. These data indicate that the plasma antioxidant system responds to ALA treatment and is correlated with tissue chemiluminescence. In vitro studies showed that ALA does not interfere with the antioxidant plasma capacity, neither promoting oxidation of plasma elements nor binding to plasma proteins.

    Topics: Amidines; Aminolevulinic Acid; Animals; Antioxidants; Ascorbic Acid; Blood Proteins; Brain; Chromans; Glutathione; Liver; Luminescent Measurements; Luminol; Male; Muscle, Skeletal; Oxidation-Reduction; Oxidative Stress; Plasma; Rats; Rats, Wistar; Sulfhydryl Compounds; Uric Acid

1997
Age-related changes in the peroxyl radical scavenging capacity of human plasma.
    Free radical biology & medicine, 1997, Volume: 23, Issue:1

    Aging and the diseases that typically follow with increasing age, notably atherosclerosis and cancer, are often proposed to be involved in increased oxidative stress. Animal studies, on the other hand, show no clear-cut pattern of age-related changes in enzymatic antioxidant defences. In this study we have demonstrated that total peroxyl radical scavenging antioxidant capacity (TRAP) in human plasma changes with age. We also found that among the antioxidants in human plasma there exists a major fraction of so far unidentified antioxidant(s). A chemiluminescent TRAP assay was used to determine the presence of peroxyl radical scavenging antioxidants in human plasma. The material consisted of 87 healthy volunteers, aged 20-96 years, who used no regular medication, vitamins, or trace elements. In females, total antioxidant capacity increased significantly during the life span. The increase in TRAP was mainly due to unidentified antioxidants. In males, TRAP increased until age 51-74, and then significantly decreased. The decrease observed among males was also due to the sharp decline in the concentration of unidentified antioxidants.

    Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aging; Animals; Antioxidants; Ascorbic Acid; Chromans; Female; Free Radical Scavengers; Free Radicals; Glutathione; Humans; Male; Middle Aged; Oxidative Stress; Peroxides; Sulfhydryl Compounds; Uric Acid; Vitamin E

1997
Cytotoxicity and apoptosis produced by arachidonic acid in Hep G2 cells overexpressing human cytochrome P4502E1.
    The Journal of biological chemistry, 1997, Jun-06, Volume: 272, Issue:23

    The goal of the current study was to evaluate the effects of arachidonic acid, as a representative polyunsaturated fatty acid, on the viability of a Hep G2 cell line, which has been transduced to express human cytochrome P4502E1 (CYP2E1). Arachidonic acid produced a concentration- and time-dependent toxicity to Hep G2-MV2E1-9 cells, which express CYP2E1, but little or no toxicity was found with control Hep G2-MV-5 cells, which were infected with retrovirus lacking human CYP2E1 cDNA. In contrast to arachidonic acid, oleic acid was not toxic to the Hep G2-MV2E1-9 cells. The cytotoxicity of arachidonic acid appeared to involve a lipid peroxidation type of mechanism since toxicity was enhanced after depletion of cellular glutathione; formation of malondialdehyde and 4-hydroxy-2-nonenal was markedly elevated in the cells expressing CYP2E1, and toxicity was prevented by antioxidants such as alpha-tocopherol phosphate, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (trolox), propylgallate, ascorbate, and diphenylphenylenediamine, and the iron chelator desferrioxamine. Transfection of the Hep G2-MV2E1-9 cells with plasmid containing CYP2E1 in the sense orientation enhanced the arachidonic acid toxicity, whereas transfection with plasmid containing CYP2E1 in the antisense orientation decreased toxicity. The CYP2E1-dependent arachidonic acid toxicity appeared to involve apoptosis, as demonstrated by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling and DNA laddering experiments. Trolox, which prevented toxicity of arachidonic acid, also prevented the apoptosis. Transfection with a plasmid containing bcl-2 resulted in complete protection against the CYP2E1-dependent arachidonic acid toxicity. It is proposed that elevated production of reactive oxygen intermediates by cells expressing CYP2E1 can cause lipid peroxidation, which subsequently promotes apoptosis and cell toxicity when the cells are enriched with polyunsaturated fatty acids such as arachidonic acid. The Hep G2-MV2E1-9 cells appear to be a valuable model to study interaction between CYP2E1, polyunsaturated fatty acids, reactive radicals, and the consequence of these interactions on cell viability and to reproduce several of the key features associated with ethanol hepatotoxicity in the intragastric infusion model of ethanol treatment.

    Topics: alpha-Tocopherol; Antioxidants; Apoptosis; Ascorbic Acid; Aspirin; Carcinoma, Hepatocellular; Cell Survival; Chromans; Cytochrome P-450 CYP2E1; Deferoxamine; Humans; L-Lactate Dehydrogenase; Liver Neoplasms; Oleic Acid; Phenylenediamines; Propyl Gallate; Proto-Oncogene Proteins c-bcl-2; Recombinant Proteins; Transfection; Tumor Cells, Cultured; Vitamin E

1997
The efficacy of antioxidants administered during low temperature storage of warm ischemic kidney tissue slices.
    Cryobiology, 1997, Volume: 34, Issue:4

    Accumulation of products of lipid peroxidation (malondialdehyde, conjugated dienes, lipid peroxides, and Schiff bases) was evaluated in rabbit kidney cortex slices made ischemic for 60 min followed by 18 h storage at 5 degrees C in UW Na gluconate solution and 210 min normothermic reoxygenated incubation. In addition, the effect of adding Trolox (1 mM), deferoxamine (1 mM), and ascorbate (1 mM) as supplemental antioxidants to the UW gluconate solution was evaluated. Lipid peroxidation was slightly increased after hypothermic storage compared to slices subjected to ischemia alone but was not significantly different than ischemic slices during subsequent incubation at normothermia. The addition of either deferoxamine or Trolox to the storage solution substantially reduced lipid peroxidation both during hypothermic storage and subsequent to normothermic incubation. Ascorbate had a mild prooxidant effect as a sole additive to the UW gluconate solution but was clearly prooxidant when combined with either deferoxamine or Trolox. These results suggest that supplemental antioxidants added to the UW gluconate solution under conditions analogous to machine perfusion preservation have a potential role in reducing oxidative stress in kidney tissues harvested after warm ischemia and that hypothermia may be a valuable adjunct to resuscitative therapeutic regimens developed for salvage of ischemic kidneys for transplantation.

    Topics: Animals; Antioxidants; Ascorbic Acid; Chromans; Cryopreservation; Deferoxamine; Ischemia; Kidney Cortex; Malondialdehyde; Rabbits; Tissue Preservation

1997
Light emission resulting from hydroxylamine-induced singlet oxygen formation of oxidizing LDL particles.
    Photochemistry and photobiology, 1997, Volume: 66, Issue:4

    Oxidation of low-density lipoprotein (LDL) by low amounts of cupric ions resulted in the formation of singlet oxygen (1O2, 1 delta g) when hydroxylamine (NH2OH) was added. Direct evidence on this excited species came from partial spectral resolution of the emitted light in the red spectral region (634 nm and 703 nm), which can be attributed to the dimol decay of singlet oxygen. Additional evidence for the existence of singlet oxygen came from the enhancing effect of deuterium oxide buffer (D2O) on chemiluminescence intensity and the quenching effect of sodium azide. A linear correlation between NH2OH-dependent chemiluminescence intensity and the amount of diene conjugates (DC) formed in this reaction was observed. Removal of adventitious transition metals by adequate chelators prevented chemiluminescence in this system; NH2OH was also found to efficiently decrease metabolites of lipid peroxidation (LPO). Our findings are consistent with a sequence of reactions in which NH2OH first converts transition metals to their reduced state, thereby stimulating the formation of alkoxy- and peroxyradicals. Peroxyradicals decompose in a bimolecular Russel reaction to hydroxyl compounds and singlet oxygen while the majority of alkoxy radicals are eliminated by a secondary reaction with NH2OH. Identical effects were observed when reducing antioxidants such as ascorbic acid or trolox C were used instead of hydroxylamine.

    Topics: Antioxidants; Ascorbic Acid; Chromans; Humans; Hydroxylamine; In Vitro Techniques; Light; Lipoproteins, LDL; Luminescent Measurements; Oxygen; Photochemistry; Singlet Oxygen

1997
The effect of simple hypothermic preservation with Trolox and ascorbate on lipid peroxidation in dog kidneys.
    Cryobiology, 1996, Volume: 33, Issue:2

    Dog kidneys were preserved by simple hypothermic storage in UW-lactobionate organ preservation solution for 48 h and analyzed for evidence of lipid peroxidation. The antioxidant function of the UW solution was evident in stored kidney tissues which had significant reductions in conjugated dienes, lipid peroxides, and Schiff base content compared to fresh controls without vascular flushing. Aerobic incubation of kidney cortex homogenates at 37 degrees C for 60 min resulted in increases in conjugated dienes, lipid peroxides, and Schiff bases in UW-stored kidneys. Schiff base production was markedly higher in UW-stored kidneys during warm incubation than in controls. Addition of Trolox (200 microM) to UW solution resulted in significant reductions in Schiff base production during warm aerobic incubation after preservation. In contrast, adding ascorbate (1 mM) to UW solution potentiated oxidative stress during aerobic incubation, with significant increases in conjugated dienes, lipid peroxides, and Schiff bases which were only partially reversed by further addition of Trolox. Increased oxidative stress was correlated with decreased respiratory function (decreased uncoupled respiration rates and sensitivity to oligomycin inhibition) in aerobically incubated homogenates. This study showed that although the UW solution does have an antioxidant function during hypothermic preservation there remains an increased oxidative stress during warm reoxygenation even in optimally harvested kidneys. The antioxidant effect of the UW solution after preservation can be significantly enhanced using the water-soluble vitamin E analogue Trolox. Antioxidant supplementation of UW solution may be advantageous in preserving kidneys with increased oxidative stresses obtained from suboptimal donors in clinical practice.

    Topics: Adenosine; Allopurinol; Animals; Antioxidants; Ascorbic Acid; Chromans; Cold Temperature; Dogs; Female; Glutathione; In Vitro Techniques; Insulin; Kidney; Lipid Peroxidation; Male; Malondialdehyde; Organ Preservation; Organ Preservation Solutions; Raffinose

1996
The reaction between ABTS radical cation and antioxidants and its use to evaluate the antioxidant status of serum samples.
    Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas, 1996, Volume: 29, Issue:2

    The 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation can be generated by incubation of ABTS and 2,2'-azo-bis(2- amidinopropane) at 45 degrees C. The ABTS radical cation is stable for several minutes at room temperature and reacts quantitatively and instantaneously with several antioxidants, such as Trolox, ascorbic acid, uric acid, cysteine, glutathione and bilirubin. In contrast, the ABTS radical cation reacts slowly with albumin. When serum is added to a solution of the ABTS radical cation, the bleaching of the radical follows biphasic kinetics, with a fast decay followed by a slow decay that takes place within several minutes. The fast decay is primarily due to uric acid, while the slow decay is related to the protein content of the sample. We propose that this procedure can provide an independent and simultaneous evaluation of the low molecular weight and protein antioxidants present in biological samples such as serum.

    Topics: Antioxidants; Ascorbic Acid; Benzothiazoles; Bilirubin; Chromans; Cysteine; Glutathione; Indicators and Reagents; Sulfonic Acids; Temperature; Time Factors; Uric Acid

1996
Anti-oxidant treatment for shock: vitamin E but not vitamin C improves survival.
    The American surgeon, 1996, Volume: 62, Issue:10

    Anti-oxidant therapy has been effective for treatment of experimental shock. In this study, the efficacy of Trolox (Aldrich Chemical Co., Milwaukee, WI), a water-soluble vitamin E analogue, and ascorbic acid (vitamin C) was evaluated in a rat model of hemorrhagic shock and resuscitation. In two prospective trials, rats were phlebotomized (27 mL/kg) and left in shock for 45 minutes. Resuscitation was then instituted by continuous IV infusion with lactated Ringer's (LR) (54 mL/kg) over 60 min. In Trial 1, rats were randomized to receive either placebo (LR) or Trolox (50 mg/kg) in LR. In Trial 2, rats were randomized to LR alone or ascorbic acid (50 mg/kg) in LR. Survival for ascorbic acid-treated rats (35 per cent) was not different than for control rats (35 per cent). However, the addition of Trolox to infusion significantly improved 72 hour survival, 75 per cent versus 40 per cent respectively, for Trolox-treated and control animals. These data demonstrate that Trolox is of survival benefit when added to resuscitation in this model. This benefit does not appear to be related to blood pressure or white cell adhesion. Trolox is more effective than ascorbic acid in this model.

    Topics: Animals; Antioxidants; Ascorbic Acid; Blood Pressure; Chromans; Disease Models, Animal; Fluid Therapy; Leukocyte Adherence Inhibition Test; Leukocytes; Prospective Studies; Rats; Rats, Sprague-Dawley; Resuscitation; Shock, Hemorrhagic; Vitamin E

1996
Protective effects of trolox C, vitamin C, and catalase on bromobenzene-induced damage to rat hepatocytes.
    Scandinavian journal of gastroenterology, 1996, Volume: 31, Issue:8

    The protective effects of trolox C (water-soluble vitamin E), vitamin C, and catalase on bromobenzene (BB)-induced toxicity to isolated rat hepatocytes were evaluated. The glutathione (GSH) content of the hepatocytes exposed to BB was measured.. BB caused acute damage to the cells during 2 h of incubation (short) when BB was added directly to the culture wells, whereas a late-occurring and time-dependent increase in lactate dehydrogenase (LDH) leakage rate was observed during 24 h of incubation (long) when BB was dissolved in a different way. Incubation of the cells with trolox C (0.5-2.0 mM) prevented the hepatocellular damage induced by BB at 2.4 mM during the long-term incubation. Vitamin C (0.1-1.0 mM) had a protective effect on BB-induced toxicity during both the short- (BB, 1.6 mM) and the long- (BB, 2.4 mM) term incubations. Catalase (3200 U/ml) also showed a beneficial effect on the cells during the short-term BB exposure. Trolox C (2.0 mM) and vitamin C (0.5 mM) restored BB-induced GSH depletion in the cells.. BB induced two patterns of LDH leakage from isolated hepatocytes on the basis of different ways of BB exposure and incubation periods. Trolox C, vitamin C, and catalase exerted protective effects on BB-induced toxicity during short- or/and long-term incubations. The effects were concentration-dependent. Restoration of GSH content in BB-exposed hepatocytes suggests that trolox C and vitamin C could reduce GSH consumption during BB metabolism and exert an antioxidant effect.

    Topics: Analysis of Variance; Animals; Antioxidants; Ascorbic Acid; Bromobenzenes; Catalase; Cells, Cultured; Chromans; Female; Glutathione; L-Lactate Dehydrogenase; Liver; Rats; Rats, Sprague-Dawley; Vitamin E

1996
Maternal infusion of antioxidants (Trolox and ascorbic acid) protects the fetal heart in rabbit fetal hypoxia.
    Pediatric research, 1996, Volume: 39, Issue:3

    The antioxidants, Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid, a water soluble analog of vitamin E) and ascorbic acid (AA), protect the heart from ischemia-reperfusion injury. We hypothesized that maternal infusion of Trolox and AA, would reduce the fetal bradycardia and myocardial damage observed in fetal hypoxia and increase the total antioxidant activity in fetal plasma. Either i.v. saline (control group) or Trolox + AA (drug group) was randomly administered to 29-d-old pregnant rabbits. Fetal hypoxia was induced by uterine ischemia. Fetal heart rate, plasma CK-MB activity, and plasma total radical antioxidant potential (TRAP) were measured in different sets of animals. Fetal heart rate in the drug group was higher than in the control group for the first 35 min (p < 0.05 at every 5-min interval). Fetal bradycardia (<60 beats/min) occurred after 39 min (median) in the drug group, and 29 min in the control group (p < 0.05). After 50 min of hypoxia, plasma CK-MB was lower in the drug group, 1204 +/- 132 U/L (mean +/- SEM), than in the control group, 2633 +/- 233 U/L (p < 0.05). TRAP was higher in the drug group, 3.01 +/- 0.15 mM (Trolox equivalent concentration), than in the control group, 1.48 +/- 0.27 mM (p < 0.05). Higher TRAP levels (> or = 2.0 mM) were associated with lower CK-MB levels (<2500 U/L) (p < 0.05). Administration of Trolox and AA to the mother has a beneficial effect on fetal myocardial damage after fetal hypoxia, and a small beneficial effect on fetal bradycardia during hypoxia. The beneficial effect may be due to the augmentation of fetal plasma antioxidants from maternal antioxidant pretreatment.

    Topics: Animals; Antioxidants; Ascorbic Acid; Bradycardia; Chromans; Creatine Kinase; Female; Fetal Hypoxia; Heart; Pregnancy; Rabbits; Reperfusion Injury; Survival

1996
Sensitivity of ATPase-ADPase activities from synaptic plasma membranes of rat forebrain to lipid peroxidation in vitro and the protective effect of vitamin E.
    Neurochemical research, 1996, Volume: 21, Issue:3

    The in vitro effects of membrane lipid peroxidation on ATPase-ADPase activities in synaptic plasma membranes from rat forebrain were investigated. Treatment of synaptic plasma membranes with an oxidant generating system (H(2)0(2)/Fe(2+)/ascorbate) resulted in lipid peroxidation and inhibition of the enzyme activity. Besides, trolox as a water soluble vitamin E analogue totally prevented lipid peroxidation and the inhibition of enzyme activity. These results demonstrate the susceptibility of ATPase-ADPase activities of synaptic plasma membranes to free radicals and suggest that the protective effect against lipid peroxidation by trolox prevents the inhibition of enzyme activity. Thus, inhibition of ATPase-ADPase activities of synaptic plasma membranes in cerebral oxidative stress probably is related to lipid peroxidation in the brain.

    Topics: Adenosine Triphosphatases; Animals; Antioxidants; Apyrase; Ascorbic Acid; Chromans; Ferrous Compounds; Hydrogen Peroxide; Kinetics; Lipid Peroxidation; Male; Prosencephalon; Rats; Rats, Wistar; Synaptic Membranes; Vitamin E

1996
Do antioxidant vitamins reduce infarct size following acute myocardial ischemia/reperfusion?
    Cardiovascular drugs and therapy, 1995, Volume: 9, Issue:1

    There is controversy concerning the ability of antioxidant vitamins to reduce myocardial infarct size. We sought to determine whether a brief prophylactic treatment of vitamin C or vitamin C plus Trolox (a water-soluble form of vitamin E) could reduce myocardial infarct size in an experimental model. We used an anesthetized open-chest rabbit model in which a branch of the circumflex coronary artery was ligated for 30 minutes followed by 4 hours of reperfusion. Experiments were performed in a randomized and blinded fashion. An IV injection of normal saline pH balanced to 7.4 (control group n = 15), vitamin C (150 mg/kg, n = 14), or vitamin C plus Trolox (150 mg/kg plus 100 mg/kg, respectively, n = 15) was administered prior to coronary occlusion. Collateral blood flow during coronary occlusion was measured by radioactive microspheres, myocardial risk zone (AR) was assessed by blue dye injection, and myocardial infarct size (AN) was assessed by triphenyltetrazolium chloride staining. All rabbits received comparable ischemic insult: Collateral blood flow and AR were similar among all three groups. Infarct size, measured as a percent of AR, did not differ significantly among the controls (21%), vitamin C (29%), or the vitamin C plus Trolox (18%) groups. Therefore, in this ischemia/reperfusion model, antioxidant vitamins did not alter myocardial infarct size.

    Topics: Animals; Antioxidants; Ascorbic Acid; Blood Pressure; Chromans; Disease Models, Animal; Drug Therapy, Combination; Free Radical Scavengers; Heart Rate; Hydrogen-Ion Concentration; Male; Myocardial Infarction; Myocardial Reperfusion Injury; Rabbits; Random Allocation; Staining and Labeling; Tetrazolium Salts; Vitamin E

1995
Detection of oxidants in uremic plasma by electron spin resonance spectroscopy.
    Kidney international, 1995, Volume: 48, Issue:1

    Depletion of antioxidants and the presence of products of free radical damage in plasma suggest that oxidative stress is increased in uremia. We have developed an application of electron spin resonance spectroscopy, and used this method to show that a stable oxidizing component or components of plasma accumulate in uremia. No oxidizing activity was detectable in plasma from subjects with normal renal function. The oxidant was detected by its capacity to oxidize the spin trap 3,5-dibromo-4-nitrosobenzene sulphonate (DBNBS). The oxidant was dialyzable from plasma, had an upper molecular weight limit of about 3,000 Daltons and was stable over many months. Physiological plasma concentrations of vitamin C, a water soluble congener of vitamin E and reduced glutathione were unable to inhibit the oxidizing capacity of uremic plasma. Thus, uremia is associated with accumulation of an endogenous oxidizing activity at much higher concentrations than in subjects with normal renal function.

    Topics: Adult; Antioxidants; Ascorbic Acid; Azides; Benzenesulfonates; Chromans; Creatinine; Electron Spin Resonance Spectroscopy; Endopeptidases; Female; Glutathione; Humans; Hydrolysis; Leukocyte Count; Male; Middle Aged; Neutrophils; Nitroso Compounds; Oxidants; Oxidation-Reduction; Renal Dialysis; Spin Labels; Uremia; Vitamin E

1995
Vitamin E, selenium, trolox C, ascorbic acid palmitate, acetylcysteine, coenzyme Q, beta-carotene, canthaxanthin, and (+)-catechin protect against oxidative damage to kidney, heart, lung and spleen.
    Free radical research, 1995, Volume: 22, Issue:2

    Male Sprague-Dawley rats were fed diets that varied qualitatively and quantitatively in antioxidants. Kidney, heart, lung, and spleen homogenates were incubated at 37 degrees C with and without hydroperoxide or Fe+2. Protection of antioxidants against oxidative damage to tissue was determined by measurement of oxidized heme proteins. Tissues from rats supplemented with dietary vitamin E and selenium showed protection compared to tissues from rats on the basal diet. Tissues from rats with diets containing larger quantities of antioxidants and both fat soluble antioxidants: vitamin E, beta-carotene, coenzyme Q10, ascorbic acid 6-palmitate and water soluble antioxidants: selenium, trolox C, acetylcysteine, coenzyme Q0, (+)-catechin, showed the highest protection.

    Topics: Acetylcysteine; Animals; Antioxidants; Ascorbic Acid; beta Carotene; Canthaxanthin; Carotenoids; Catechin; Chromans; Heart; Liver; Lung; Male; Myocardium; Oxidative Stress; Rats; Rats, Sprague-Dawley; Selenium; Spleen; Ubiquinone; Vitamin E

1995
Protection of vitamin E, selenium, trolox C, ascorbic acid palmitate, acetylcysteine, coenzyme Q0, coenzyme Q10, beta-carotene, canthaxanthin, and (+)-catechin against oxidative damage to rat blood and tissues in vivo.
    Free radical biology & medicine, 1995, Volume: 18, Issue:5

    Male Sprague-Dawley rats were fed either a vitamin E and selenium deficient diet, a diet supplemented with vitamin E and selenium, or a diet supplemented with vitamin E, selenium, trolox C, ascorbic acid palmitate, acetylcysteine, Beta-carotene, canthaxanthin, coenzyme Q0, coenzyme Q10, and (+)-catechin. Rats were injected with CBrCl3 (0.05 mmol/100 g body weight) intraperitoneally. Oxidative damage to tissues was measured by formation of oxidized heme proteins (OHP) in blood, liver, kidney, heart, lung, and spleen. Diets supplemented with antioxidants showed protection against oxidative damage caused by CBrCl3. The protection was dependent on the diversity and quantity of antioxidants in the diet. In general, diets supplemented with both fat soluble and water soluble antioxidants provided better protection than diets supplemented only with vitamin E and selenium or with vitamin E, selenium, and fat soluble antioxidants.

    Topics: Acetylcysteine; Animals; Antioxidants; Ascorbic Acid; beta Carotene; Canthaxanthin; Carotenoids; Catechin; Chromans; Coenzymes; Heart; Hemeproteins; Kidney; Liver; Lung; Male; Myocardium; Oxidation-Reduction; Oxidative Stress; Palmitic Acid; Palmitic Acids; Rats; Rats, Sprague-Dawley; Selenium; Spleen; Ubiquinone; Vitamin E; Vitamin E Deficiency

1995
Electroporation enhances cell membrane peroxidation and luminescence.
    Biochemical and biophysical research communications, 1995, Jan-05, Volume: 206, Issue:1

    Electroporation is a most popular method of cell membrane permeabilization, by pulsed electric fields. It allows foreign molecules to enter the cell and has been used for many biotechnological applications, including transformation of mammalian cells and plant protoplasts by exogenous genetic material. However, the mechanism underlying membrane electropermeabilization is still largely unknown. Evidence is presented here that electroporation under conditions compatible with cell survival induces lipid hydroperoxide formation in the membranes of animal and plant cells. Exposure to electric fields also enhanced up to 5-fold the spontaneous emission of light from both cell types, which paralleled the amount of conjugated hydroperoxides detected in cell membranes. The emitted photons were mainly in the red edge of the spectrum, suggesting the involvement of singlet oxygen. The presence of antioxidants during electroporation did not reduce the formation of hydroperoxides nor the permeability but quenched the luminescence.

    Topics: Antioxidants; Ascorbic Acid; Cell Membrane; Chromans; Electroporation; Fabaceae; Humans; Leukemia, Erythroblastic, Acute; Lipid Peroxidation; Luminescent Measurements; Membrane Lipids; Oxygen; Photochemistry; Plants, Medicinal; Protoplasts; Singlet Oxygen; Tumor Cells, Cultured; Vitamin E

1995
A fluorescence-based method for measuring total plasma antioxidant capability.
    Free radical biology & medicine, 1995, Volume: 18, Issue:1

    The Total Radical-Trapping Antioxidant Parameter (TRAP) of 10 freshly prepared human plasmas was measured by a new fluorometric assay. In this method, the rate of peroxidation induced by 2,2'-diazobis (2-amidinopropane) dihydrochloride (ABAP) was monitored through the loss of fluorescence of the protein R-Phycoerythrin (R-PE). The lag-phase induced by plasma was compared to that induced by 6-hydroxy-2,5,7,8-tetramethylchroman-2- carboxylic acid (Trolox, a water-soluble analogue of vitamin E). Proteins (but not their sulphydryl groups) interfere with the analysis, partially protecting R-PE when all plasma antioxidants are exhausted. A Trolox-induced lag-phase must therefore be measured on each plasma sample. We found that ascorbate (2.5-5.3%), alpha-tocopherol (2.9-8.5%), urate (19.6-61.0), and thiol groups (17.3-42.3%) jointly explain up to 70% of TRAP. Thus, either other compounds present in plasma are likely to exert antioxidant action, or a marked synergistic action between antioxidants should be postulated to exist. This latter hypothesis is supported by the finding that the simultaneous inactivation of ascorbate and thiol groups produces a loss in antioxidant capacity of plasma greater (26%) than the sum of the decreases produced by the separate inactivation of each of the two compounds. The proposed method appears simple, reliable, and allows the rapid handling of a reasonable number of freshly prepared plasma samples. Given the rapid loss of TRAP upon storage, the latter characteristic is crucial in studies on humans, involving a large number of subjects.

    Topics: Adult; Amidines; Antioxidants; Ascorbic Acid; Chromans; Free Radicals; Humans; Peroxides; Phycoerythrin; Spectrometry, Fluorescence; Sulfhydryl Compounds; Uric Acid; Vitamin E

1995
Comparative cytoprotection of cultured corneal endothelial cells by water-soluble antioxidants against free-radical damage.
    Cornea, 1995, Volume: 14, Issue:5

    We reported previously that purpurogallin (PPG) markedly protects the cultured rabbit corneal endothelial cells (RCEC) against oxyradical damage generated with hypoxanthine (HX) and xanthine oxidase (XO)(1). In this study, we further compared the cytoprotective activities of PPG versus Trolox (TX, alpha-tocopherol, a water-soluble analogue of vitamin E) and ascorbate (Asc) in confluent cultured RCEC with phase contrast microscopy and confirmed by transmission electron microscopy. PPG prolonged survival of the oxyradical damaged cells longer than those without PPG present (18.6 +/- 1.4 min at 1.0 mM and 11.2 +/- 1.0 at 0.25 mM respectively vs. 7.3 +/- 0.8 min in control). At levels equimolar to PPG, TX, and Asc were less effective in delaying cell necrosis caused by HX and XO (p < 0.01). When exposed to superoxide radicals generated by menadione, RCEC necrosed at 29.8 +/- 1.5 min compared to PPG 47.2 +/- 1.0 min at 1.0 mM and 38.9 +/- 1.0 min at 0.25 mM. This was significantly different from TX and Asc at corresponding concentrations (p < 0.01). PPG scavenges not only HX-XO-generated oxyradicals, but also nonenzymatically produced superoxide radicals, more actively than two well known antioxidants--TX and Asc.

    Topics: Animals; Antioxidants; Ascorbic Acid; Benzocycloheptenes; Cell Death; Cell Survival; Cells, Cultured; Chromans; Endothelium, Corneal; Free Radical Scavengers; Free Radicals; Hypoxanthine; Hypoxanthines; Rabbits; Reactive Oxygen Species; Solubility; Vitamin K; Xanthine Oxidase

1995
Differential effects of in vitro peroxidation on peripheral- and central-type benzodiazepine receptors. Protection by diverse antioxidants.
    Biochemical pharmacology, 1995, Nov-27, Volume: 50, Issue:11

    The influence of various concentrations of ferrous iron and ascorbate on in vitro peroxidation and drug binding of diverse membrane preparations (cerebral cortex and liver) was studied. Peroxidation was not simply dose-related to ascorbate and ferrous iron, but a complex relationship between iron and ascorbate when added in association was established. Under our conditions 0.01 mM Fe2+ and 0.5 mM ascorbate was the most peroxidative combination for cerebral and liver membranes. Under the same conditions, cerebral membranes were more peroxidated than liver membranes. Considering the consequences of drug binding, peripheral-type benzodiazepine receptors (PBRs) of liver were more affected by peroxidative events than central-type benzodiazepine receptors (CBRs) of the cerebral cortex. The degree of binding disturbance was generally inversely correlated to the degree of peroxidation and this was more significant for liver PBRs than for cerebral CBRs. The liver membrane model was retained for testing in vitro protection by diverse putative antioxidants. Under our conditions desferrioxamine, ethylene diamine tetra acetate (EDTA), trolox, and rutin were good protective antioxidants, whereas phenyl-butyl-nitrone (PBN) and tocopherol were not effective.

    Topics: Animals; Antioxidants; Ascorbic Acid; Benzodiazepinones; Brain; Cell Membrane; Chromans; Deferoxamine; Edetic Acid; Ferrous Compounds; Flunitrazepam; Hypolipidemic Agents; Lipid Peroxidation; Liver; Malondialdehyde; Receptors, GABA-A; Rutin; Vitamin E

1995
Trolox protects mitochondrial complex IV from nitric oxide-mediated damage in astrocytes.
    Brain research, 1994, Dec-30, Volume: 668, Issue:1-2

    The efficacy of cystine, ascorbate and trolox, a vitamin E analogue, at protecting against nitric oxide-mediated mitochondrial complex IV damage has been investigated in cultured astrocytes. Of these compounds, only trolox afforded protection. It is suggested that lipid peroxidation is responsible for nitric oxide-mediated mitochondrial damage and that inhibitors of this process may be of therapeutic benefit in conditions where excessive nitric oxide production is implicated.

    Topics: alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; Amino Acid Oxidoreductases; Animals; Ascorbic Acid; Astrocytes; Chromans; Cystine; Electron Transport Complex IV; Enzyme Induction; Interferon-gamma; Lipid Peroxidation; Lipopolysaccharides; Mitochondria; Nitric Oxide; Nitric Oxide Synthase; Rats; Rats, Wistar

1994
Visible chemiluminescence associated with the reaction between methemoglobin or oxyhemoglobin with hydrogen peroxide.
    Photochemistry and photobiology, 1994, Volume: 60, Issue:5

    Visible chemiluminescence is emitted in the irreversible deactivation of hemoglobin or methemoglobin with excess H2O2. The emission takes place in two phases. The most intense one lasts a few seconds and is followed by a second phase of lower intensity that remains for longer periods. This second phase presents chaotic or sustained oscillations. Free radicals are implicated in the luminescent process since the emission can be reduced by free radical scavengers such as 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) or ascorbic acid. These additives lead to a delay in reaching the maximum intensity, which can be related to their consumption, implying substantial recycling of the hemoprotein. Chemiluminescence is also observed in the oxidation of hemin by H2O2, suggesting a role for the heme group in the processes leading to the excited state production. The lower intensity observed in the presence of hemin can be related to the contribution of the globin chains.

    Topics: Animals; Antioxidants; Ascorbic Acid; Cattle; Chromans; Free Radical Scavengers; Free Radicals; Hydrogen Peroxide; Luminescent Measurements; Methemoglobin; Oxyhemoglobins

1994
[Induction and prevention of kidney injury induced by GSH depletion and vitamin E deficiency in rats].
    Nihon Jinzo Gakkai shi, 1994, Volume: 36, Issue:10

    Four-week-old Wistar male rats were fed a vitamin E (VE)-deficient diet for 8 weeks, followed by intraperitoneal injection of DL-buthionine- [S, R] -sulfoximine (BSO), an inhibitor of gamma-glutamylcysteine synthetase, at the dose of 1 mmol/kg body weight. As we reported previously, GSH depletion by administration of BSO induced acute tubular necrosis in the kidney of VE-deficient rats and was accompanied by decrease of renal TBA value and marked increase of renal lipofuscin content. In this study, we examined the effect of administration of AsA or Trolox C on these kidney injuries. AsA or Trolox C treatment increased renal GSH content and inhibited the increase of renal lipofuscin production. The increase of BUN and creatinine levels and LDH activity in the sera of rats administered BSO were inhibited by AsA or Trolox C treatment. AsA treatment completely protected the necrosis of epithelia of proximal renal tubules. These results suggest that GSH has an important role in preventing lipofuscin production through the reaction of lipid peroxides with amino acids. AsA spares GSH indicating that these compounds have similar antioxidant actions and that AsA can serve as an essential antioxidant in the presence of severe GSH deficiency.

    Topics: Animals; Ascorbic Acid; Buthionine Sulfoximine; Chromans; Glutathione; Kidney Tubular Necrosis, Acute; Male; Methionine Sulfoximine; Rats; Rats, Wistar; Vitamin E; Vitamin E Deficiency

1994
Protection by vitamin E selenium, trolox C, ascorbic acid palmitate, acetylcysteine, coenzyme Q, beta-carotene, canthaxanthin, and (+)-catechin against oxidative damage to liver slices measured by oxidized heme proteins.
    Free radical biology & medicine, 1994, Volume: 16, Issue:4

    Male SD rats were fed a vitamin E- and selenium-deficient diet, a diet supplemented with vitamin E and selenium, and diets supplemented with vitamin E, selenium, trolox C, ascorbic acid palmitate, acetylcysteine, beta-carotene, canthaxanthin, coenzyme Q0, coenzyme Q10, and (+)-catechin. Liver slices were incubated at 37 degrees C with and without CBrCl3, t-butyl-hydroperoxide, Fe+2, or Cu+2. The effect of antioxidant nutrients on the oxidative damage to rat liver was studied by measurement of the production of oxidized heme proteins (OHP) during the oxidative reactions. Diet supplemented with vitamin E and selenium showed a strong protection against heme protein oxidation compared to the antioxidant-deficient diet. Furthermore, increasing the diversity and quantity of antioxidants in the diets provided significantly more protection.

    Topics: Acetylcysteine; Animals; Antioxidants; Ascorbic Acid; beta Carotene; Canthaxanthin; Carotenoids; Catechin; Chromans; Dose-Response Relationship, Drug; Hemeproteins; In Vitro Techniques; Male; Oxidants; Oxidation-Reduction; Rats; Rats, Sprague-Dawley; Selenium; Ubiquinone; Vitamin D Deficiency; Vitamin E

1994
Morin: a wood pigment that protects three types of human cells in the cardiovascular system against oxyradical damage.
    Biochemical pharmacology, 1994, Mar-15, Volume: 47, Issue:6

    Morin is a yellowish pigment extractable from the wood of Chlorophora tinctoria. In the present study, we have determined that morin protects three types of human cells--ventricular myocytes, saphenous vein endothelial cells, and erythrocytes--against damage by oxyradicals generated in situ. In myocytes and endothelial cells, morin prolonged substantially and in a concentration-dependent manner the survival of cells exposed to either xanthine oxidase-generated oxyradicals or superoxide radicals produced with menadione. Morin protected erythrocytes from lytic attack by peroxyl radicals generated with 2,2'-azo-bis (2-amidinopropane) dihydrochloride. In all three types of human cells, the protective effect of morin clearly excelled that displayed by Trolox (a vitamin E analog), ascorbate, or mannitol, which are water-soluble antioxidants of similar molecular size. Chemically, we verified that morin behaves as an antioxidant by diminishing markedly the amount of malondialdehyde (lipid peroxidation product) found in human cardiocytes despite their exposure to oxyradicals. In agreement with related reports, we also observed that morin is non-toxic in rats even when used at concentrations 2-3 orders of magnitude higher than those in our in vitro studies. Thus, morin acts as a broad-spectrum and non-toxic antioxidant.

    Topics: Ascorbic Acid; Cells, Cultured; Chromans; Endothelium, Vascular; Erythrocytes; Flavonoids; Free Radical Scavengers; Heart Ventricles; Humans; Malondialdehyde; Mannitol; Reactive Oxygen Species; Vitamin K; Wood

1994
Inhibition of hydroxyl radical-induced protein damages by trolox.
    Biochemistry and molecular biology international, 1993, Volume: 31, Issue:1

    When bovine serum albumin was exposed to the hydroxyl radical generating system of ascorbate-EDTA-Fe3+ or ascorbate-EDTA-Fe(3+)-H2O2, carbonyl formation occurred. Trolox strongly inhibited the oxidative modification in a dose-dependent manner. The fragmentation and enhancement of the proteolytic susceptibility of bovine serum albumin were induced by HO. only in the presence of O2, while trolox protected the protein against such oxidative modifications. In addition, inactivations of lactate dehydrogenase and creatine kinase induced by the hydroxyl radical were also blocked by trolox.

    Topics: Aerobiosis; Anaerobiosis; Antioxidants; Ascorbic Acid; Chromans; Creatine Kinase; Dose-Response Relationship, Drug; Edetic Acid; Electrophoresis, Polyacrylamide Gel; Ferric Compounds; Hydrogen Peroxide; Hydroxyl Radical; L-Lactate Dehydrogenase; Oxidation-Reduction; Proteins; Serum Albumin, Bovine

1993
Ginkgo biloba extract EGb 761 or trolox C prevent the ascorbic acid/Fe2+ induced decrease in synaptosomal membrane fluidity.
    Free radical research communications, 1993, Volume: 19, Issue:5

    The ability of synaptosomes, prepared from striata, to take up 3H-dopamine declined rapidly during incubation at 37 degrees C, in an oxygenated Krebs-Ringer medium with 0.1 mM ascorbic acid. Ascorbic acid was responsible for this decrease. Its effectiveness after a 60 min incubation was concentration dependent from 1 microM and virtually complete for 0.1 mM. Furthermore, a decrease of synaptosomal membrane fluidity was revealed by measurements of fluorescence polarization using 1,6-diphenyl-1,3,5-hexatriene. This decrease was potentiated by Fe2+ ions (1 microM). In contrast, it was prevented by the Fe2+ ion chelator, desferrioxamine (0.1 mM), by the Ginkgo biloba extract EGb 761 [2-16 micrograms/ml], as well as by the flavonoid quercetin (0.1 microM). This preventive effect was shared by trolox C (from 0.1 mM). It is concluded that peroxidation of neuronal membrane lipids induced by ascorbic acid/Fe2+ is associated with a decrease in membrane fluidity which, in turn, reduces the ability of the dopamine transporter to take up dopamine.

    Topics: Animals; Antioxidants; Ascorbic Acid; Chromans; Diphenylhexatriene; Dopamine; Dose-Response Relationship, Drug; Ferrous Compounds; Fluorescence; Furans; Ginkgo biloba; In Vitro Techniques; Male; Membrane Fluidity; Mice; Plant Extracts; Quercetin; Synaptic Membranes

1993
Rates of interactions of superoxide with vitamin E, vitamin C and related compounds as measured by chemiluminescence.
    Biochimica et biophysica acta, 1992, Jan-23, Volume: 1115, Issue:3

    The rate constants for the interactions of superoxide with vitamin E (alpha-tocopherol), vitamin C (ascorbic acid) and their related compounds have been measured by a chemiluminescence method. A strong chemiluminescence of a constant intensity was observed when xanthine oxidase was added to an aqueous solution of hypoxanthine and a Cypridina luciferin analog, 2-methyl-6-phenyl-3-7-dihydroimidazo[1,2-a]pyrazin-3-one (CLA). Vitamin E, vitamin C and their related compounds competed with CLA to react with superoxide and reduced the chemiluminescence intensity. From a kinetic analysis of the effect of addition of these compounds on the chemiluminescence intensity, the rate constants for their interactions with superoxide were measured at 25 degrees C and pH 7.8. The rate constants were obtained as 3.3 x 10(5) and 1.7 x 10(4) M-1 s-1 for ascorbate and 2-carboxy-2,5,7,8-tetramethyl-6-chromanol, respectively, and also as 4.9 x 10(3) and 4.5 x 10(3) M-1 s-1 for alpha-tocopherol incorporated into soybean and dimyristoyl phosphatidylcholine liposomal membranes, respectively. It has been shown that this method is a sensitive and a quick method which can be applied for measurement of the reactivities of various natural and synthetic compounds toward superoxide. In addition it has been shown that this method can also be applied to the heterogeneous system as well as homogeneous solution, which makes it more versatile and useful for the study in biochemistry.

    Topics: Ascorbic Acid; Binding, Competitive; Chromans; Hydrogen-Ion Concentration; Hypoxanthine; Hypoxanthines; Kinetics; Liposomes; Luminescent Measurements; Phosphatidylcholines; Pyrazines; Superoxides; Vitamin E; Xanthine Oxidase

1992
Oxidation mechanism of vitamin E analogue (Trolox C, 6-hydroxy-2,2,5,7,8-pentamethylchroman) and vitamin E by horseradish peroxidase and myoglobin.
    Archives of biochemistry and biophysics, 1992, Volume: 299, Issue:2

    The oxidation of 6-hydroxy-2,2,5,7,8-pentamethylchroman, Trolox C, and alpha-tocopherol by horseradish peroxidase was examined by stopped-flow and ESR experiments. The catalytic intermediate of horseradish peroxidase during the oxidation of vitamin E analogues and vitamin E was invariably Compound II, and rate constants for the rate-determining step decreased in the order 6-hydroxy-2,2,5,7,8-pentamethylchroman > Trolox C > alpha-tocopherol. The formation of phenoxyl radicals from substrates was verified with ESR and was followed optically. Resulting 6-hydroxy-2,2,5,7,8-pentamethylchroman and Trolox C radicals decayed through a dismutation reaction, followed by formation of the quinoid form via a transient intermediate. The sequence of events after formation of 6-hydroxy-2,2,5,7,8-pentamethylchroman and Trolox C radicals was similar to that observed by pulse radiolysis (Thomas, M. J., and Bielski, B. H. J. (1989). J. Am. Chem. Soc. 111, 3315-3319). Final oxidation products of 6-hydroxy-2,2,5,7,8-pentamethylchroman and Trolox C were identified as the quinoid forms and were obtained quantitatively whether or not the analogue had a carboxyl or methyl group at the 2-position of chroman ring. In contrast, enzymatic oxidation of alpha-tocopherol gave alpha-tocopherol quinone in very low yield. Conversion of 6-hydroxy-2,2,5,7,8-pentamethylchroman, Trolox C, and alpha-tocopherol to the corresponding quinones was also catalyzed by metmyoglobin in a reaction completely inhibited by ascorbate.

    Topics: Animals; Ascorbic Acid; Chromans; Electron Spin Resonance Spectroscopy; Free Radicals; Horseradish Peroxidase; Horses; In Vitro Techniques; Kinetics; Myoglobin; Oxidation-Reduction; Spectrum Analysis; Vitamin E

1992
Effect of antioxidants on hypoxia/reoxygenation-induced injury in isolated perfused rat liver.
    Pharmacology & toxicology, 1992, Volume: 71, Issue:4

    Isolated perfused livers from rats fasted overnight were subjected to 30 min. of hypoxia followed by reoxygenation for 60 min., resulting in marked cytotoxicity as evidenced by an enhanced release of cytosolic enzymes (lactate dehydrogenase: 14-fold over controls, glutamate-pyruvate-transaminase: 12-fold over controls) and glutathione (twofold over controls) into the perfusate, by calcium accumulation (by a factor of 1.4) in the tissue and by an 80% inhibition of bile secretion. Virtually no mitochondrial injury became apparent and no evidence for lipid peroxidation could be found. In the presence of ascorbate, an augmentation of hepatic injury was observed. This might be due to the pro-oxidant activity of ascorbate in the presence of ionized iron, which is easily released from high molecular weight stores under reductive (e.g. hypoxic) conditions. The water soluble vitamin E analogue trolox C as well as propyl gallate clearly protected the liver against hypoxia/reoxygenation injury, yielding further evidence for a causative role of oxidative stress in this model. Due to their water solubility and their high efficacy as free radical scavengers, these antioxidants might be of therapeutic value.

    Topics: Animals; Antioxidants; Ascorbic Acid; Bile; Cell Hypoxia; Chromans; Disease Models, Animal; Glutamate Dehydrogenase; Glutathione; L-Lactate Dehydrogenase; Liver; Male; Oxygen Consumption; Propyl Gallate; Rats; Rats, Wistar

1992
Prevention by Ginkgo biloba extract (EGb 761) and trolox C of the decrease in synaptosomal dopamine or serotonin uptake following incubation.
    Biochemical pharmacology, 1992, Dec-15, Volume: 44, Issue:12

    Prolonged incubation of synaptosomes in Krebs-Ringer oxygenated medium in the presence of ascorbic acid (10(-4) M) led, after 20 min, to a decrease in [3H]dopamine (DA) (synaptosomes prepared from the striatum) and [3H]serotonin (5HT) (synaptosomes prepared from the cortex) uptake. The decrease was progressive and uptake was virtually abolished after a 60 min incubation period. A concentration-dependent (from 5 x 10(-6) M) role of ascorbic acid in the decrease of [3H]DA or [3H]5HT uptake was demonstrated. This decrease was potentiated by Fe2+ ions and prevented by the ferrous chelating agent desferrioxamine. Thus, the progressive decrease in synaptosomal uptake of either [3H]DA or [3H]5HT could depend on the generation of free radicals by the association of ascorbic acid with Fe2+ ions. The decrease in synaptosomal uptake was prevented, in a concentration-dependent manner, by the Ginkgo biloba extract EGb 761 (4-16 micrograms/mL) and the vitamin E analog trolox C (10(-4) M). The terpenic fraction of EGb 761, Bn 52063 (up to 0.5 microgram/mL), did not prevent the reduction of [3H]amine uptake. In contrast, the flavonoidic fraction, Cp 202, was effective (from 1 microgram/mL) and its efficacy was shared by the flavonoid quercetin (from 0.1 microgram/mL). The prolongation of the ability of synaptosomes to take up [3H]amine elicited by EGb 761, in particular its flavonoidic fraction, as well as by trolox C could be due to their free radical scavenger properties.

    Topics: Animals; Ascorbic Acid; Chromans; Corpus Striatum; Deferoxamine; Dopamine; Drug Interactions; Ginkgo biloba; In Vitro Techniques; Male; Mice; Plant Extracts; Quercetin; Serotonin; Synaptosomes; Tritium

1992
Inhibition of mammalian 5-lipoxygenase and cyclo-oxygenase by flavonoids and phenolic dietary additives. Relationship to antioxidant activity and to iron ion-reducing ability.
    Biochemical pharmacology, 1991, Oct-09, Volume: 42, Issue:9

    We investigated the ability of various plant flavonoids (a) to inhibit 5-lipoxygenase and cyclooxygenase activities in rat peritoneal leukocytes, (b) to inhibit lipid peroxidation in rat liver microsomes, and (c) to stimulate DNA degradation caused by the antibiotic bleomycin in the presence of ferric ions. These compounds were compared with a range of synthetic phenolic substances including carnosol, vanillin, vitamin E and its analogue trolox c. The flavonoids were potent inhibitors of non-enzymatic peroxidation in membranes but this was not significantly correlated with their ability to inhibit either pathway of eicosanoid synthesis, suggesting that their mode of inhibition of 5-lipoxygenase/cyclooxygenase is not simply due to interception of peroxyl radicals generated at the active site of the enzymes. Many of the flavonoids and other compounds (including carnosol, vitamin E and trolox c) stimulated Fe3+/bleomycin-dependent DNA degradation. Those flavonoids which stimulated DNA degradation at low concentrations but which inhibited it at higher concentrations ("biphasic" effect, possibly caused by changing relative contributions of ability to reduce ferric-bleomycin or to chelate iron ions from the bleomycin) were selective inhibitors of 5-lipoxygenase compared to cyclo-oxygenase. In contrast, those flavonoids that did not stimulate DNA degradation at all proved to be cyclo-oxygenase selective inhibitors. Compounds that increased Fe3+/bleomycin-dependent DNA damage up to a maintained plateau were non-selective inhibitors of both 5-lipoxygenase and cyclo-oxygenase. Thus, a combination of iron-chelating and iron ion-reducing properties appears to be required for selective 5-lipoxygenase inhibition by phenolic compounds. Carnosol, vitamin E and trolox c were also found to be 5-lipoxygenase inhibitors of varying potency, and all were less active as cyclo-oxygenase inhibitors.

    Topics: Abietanes; Animals; Antioxidants; Ascorbic Acid; Chromans; Cytochrome c Group; Diet; DNA Damage; Female; Ferric Compounds; Flavonoids; Flavonols; Leukocytes; Lipid Peroxidation; Lipoxygenase Inhibitors; Microsomes, Liver; Oxidation-Reduction; Oxygenases; Phenanthrenes; Phenols; Plant Extracts; Quercetin; Rats; Rats, Inbred Strains; Vitamin E

1991
On the ability of lactoperoxidase to catalyze the peroxidase-oxidase oxidation of a vitamin E water-soluble derivative (Trolox C).
    Biochemistry international, 1991, Volume: 25, Issue:5

    The rapid-scan spectral technique has been applied to test conversion of the lactoperoxidase compounds during the peroxidase-oxidase catalyzed oxidation of Trolox. The results clearly indicate a normal peroxidatic pathway of Trolox degradation. Changes of spectral scan profiles were investigated to study directly the interaction of Trolox with lactoperoxidase compound III. Oxygen radicals were not involved in peroxidase-mediated oxidation of Trolox. The rate of the one-electron reduction of lactoperoxidase compound I to II was the same in the absence and presence of equal amounts of Trolox and ascorbic acid, which is also a good substrate for lactoperoxidase. The oxidation of Trolox by lactoperoxidase has potential physiological relevance. Since it could help maintain the catalytic cycles and activity of animal peroxidases, leading to detoxification of hydrogen peroxide as a main product of inflammation processes.

    Topics: Ascorbic Acid; Chromans; Hydrogen Peroxide; Lactoperoxidase; Lipid Peroxidation; Oxidation-Reduction; Oxidoreductases; Peroxidase; Superoxide Dismutase

1991
Effect of antioxidants on vanadate-induced toxicity towards isolated perfused rat livers.
    Toxicology, 1991, Volume: 70, Issue:2

    The effect of trolox C, a water soluble vitamin E analogue, propyl gallate and ascorbate on vanadate hepatotoxicity was investigated in vitro. In isolated perfused livers from fasted rats, sodium orthovanadate (2 mmol/l) led to toxic responses including reduction of oxygen consumption, release of cytosolic (glutamate-pyruvate-transaminase (GPT) and lactate dehydrogenase (LDH)) and mitochondrial (glutamate-dehydrogenase (GLDH)) enzymes, intracellular accumulation of calcium, a marked depletion of glutathione (GSH) and an enhanced formation and release of thiobarbituric acid- (TBA) reactive material. Trolox C and propyl gallate inhibited the release of GPT and LDH partially and that of GLDH totally, but had no influence on vanadate-induced calcium accumulation or on the reduction of oxygen consumption. Both agents suppressed vanadate-induced lipid peroxidation (LPO) and partially prevented GSH depletion. Ascorbate failed to provide any protection probably due to the interference of its pro-oxidant potential with its antioxidant activity. The protection, mainly of mitochondria, afforded by those agents which also inhibited LPO substantiates our previous findings that the pro-oxidant activity of vanadate is mainly responsible for its direct hepatotoxic actions [2]. Besides, reduction of organ perfusion rate due to vasoconstriction also contributes to vanadate toxicity, but oxidative stress is not involved in this indirect toxic activity.

    Topics: Alanine Transaminase; Animals; Antioxidants; Ascorbic Acid; Calcium; Chromans; L-Lactate Dehydrogenase; Liver; Male; Oxygen Consumption; Propyl Gallate; Rats; Rats, Inbred Strains; Vanadates

1991
One-electron oxidation of Trolox C (a vitamin E analogue) by peroxidases.
    Journal of biochemistry, 1990, Volume: 108, Issue:2

    The oxidation mechanism of Trolox C (a vitamin E analogue) by peroxidases was examined by stopped flow and ESR techniques. The results revealed that during the oxidation of Trolox C, peroxidase Compound II was the catalytic intermediate. The rate constants for the reaction of Compound II with Trolox C, which should be the rate-determining step, were estimated to be 2.1 X 10(4) and 7.2 X 10(3) M-1.s-1 for horseradish peroxidase and lactoperoxidase, respectively, at pH 6.0. The formation of the Trolox C radical was followed by ESR. The time course of the signal was similar to that of the optical absorbance changes at 440 nm, assigned as the peak of the Trolox C radical. The signal exhibited a hyperfine structure characteristic of phenoxyl radicals. From an estimation of the radical concentration in the steady state and the velocity of the radical formation, the dismutation constant was calculated to be 5 X 10(5) M-1.s-1. The concentration of the signal in the steady state was reduced by the addition of GSH. The spectrum changed from that of the Trolox C radical to that of the ascorbate radical when the reaction was carried out in the presence of ascorbate.

    Topics: Ascorbic Acid; Chromans; Electron Spin Resonance Spectroscopy; Free Radicals; Horseradish Peroxidase; Hydrogen Peroxide; Kinetics; Lactoperoxidase; Oxidation-Reduction; Peroxidases; Vitamin E

1990
Ovothiols as free-radical scavengers and the mechanism of ovothiol-promoted NAD(P)H-O2 oxidoreductase activity.
    Biochemistry, 1990, Feb-20, Volume: 29, Issue:7

    Racemic ovothiol A [(+/-)-1a] and the ovothiol model compound 1,5-dimethyl-4-mercaptoimidazole (DMI, 2) were found to scavange the free radicals Fremy's salt (4) and Banfield' radical (5) much more rapidly than did the thiol antioxidant glutathione. Ovothiol A also scavenges the tyrosyl radical, with efficiency comparable to that of ascorbic acid and the tocopherol analogue trolox (3). The ovothiol model compound DMI was found to scavenge superoxide with a rate constant comparable to that of the reaction between superoxide and glutathione. These results suggest both a free-radical scavenging role for the ovothiols and a mechanism by which the ovothiols confer NAD(P)H-O2 oxidoreductase activity upon the enzyme ovoperoxidase. Investigation of this mechanism implicates the ovothiol thiyl radical and the NAD radical as key intermediates. The ovothiyl radical appears to be unreactive toward oxygen but highly reactive toward NADH. An estimate of the one-electron oxidation potential of the ovothiol anion is presented. The physical basis for the stability of the ovothiol free radical is discussed.

    Topics: Amino Acids, Sulfur; Antioxidants; Ascorbic Acid; Chromans; Free Radicals; Glutathione; Indicators and Reagents; Kinetics; Methylhistidines; NADH, NADPH Oxidoreductases; NADPH Oxidases; Nitroso Compounds; Structure-Activity Relationship; Sulfhydryl Compounds

1990
Myocardial salvage with trolox and ascorbic acid for an acute evolving infarction.
    The Annals of thoracic surgery, 1989, Volume: 47, Issue:4

    Both Trolox (a water-soluble analogue of alpha-tocopherol) and ascorbic acid were more effective than superoxide dismutase or catalase in protecting myocyte cell cultures from free radical attack (induced by hypoxanthine and xanthine oxidase). In a canine model of two hours of left anterior descending coronary artery occlusion followed by four hours of reperfusion, Trolox and ascorbic acid reduced the area of infarction within the area at risk. The Trolox group received 500 mL of deoxygenated saline solution containing 2.0 g of Trolox, 3.0 g of ascorbic acid, and 18 mg of EDTA (ethylenediaminetetraacetic acid) infused into the ascending aorta 30 seconds before and four minutes after reperfusion. Saline controls received 500 mL of deoxygenated saline solution containing 18 mg of EDTA. The angioplasty group had unmodified reperfusion by simple release of the occlusion. The area at risk and the area infarcted were estimated with Evans blue and triphenyl tetrazolium hydrochloride stains, respectively. The ratio of the area infarcted to the area at risk was significantly lower with Trolox (angioplasty, 30.4% +/- 5.1%; saline, 20.8% +/- 2.9%; and Trolox, 8.7% +/- 4.0%; p less than 0.01). In summary, the antioxidants Trolox and ascorbic acid effectively reduced myocardial necrosis after ischemia.

    Topics: Animals; Antioxidants; Ascorbic Acid; Benzopyrans; Cells, Cultured; Chromans; Disease Models, Animal; Dogs; Free Radicals; Heart; Hemodynamics; Myocardial Infarction; Myocardial Reperfusion Injury; Myocardium; Necrosis

1989
A qualitative fluorescence-based assay for tyrosyl radical scavenging activity: ovothiol A is an efficient scavenger.
    Analytical biochemistry, 1989, Aug-01, Volume: 180, Issue:2

    A method for determining relative tyrosyl radical scavenging activity of antioxidants which requires only a standard fluorometer and commercially available materials is presented. Ultraviolet irradiation of aqueous tyrosine solutions containing superoxide dismutase and catalase produces fluorescent dityrosine residues via dimerization of photogenerated tyrosyl radicals. Added antioxidants suppress the buildup of fluorescence by scavenging the tyrosyl radicals. A correlation exists between the ability of a substance to suppress dityrosine formation and the substance's one-electron oxidation potential. This method demonstrates that ovothiol A scavenges tyrosyl radicals much more efficiently than glutathione or cysteine, resembling instead the known biological radical scavengers uric acid and ascorbic acid and the alpha-tocopherol analog trolox.

    Topics: Animals; Antioxidants; Ascorbic Acid; Cattle; Chromans; Free Radicals; Methylhistidines; Spectrometry, Fluorescence; Tyrosine; Uric Acid

1989
Vitamin E analogue Trolox C. E.s.r. and pulse-radiolysis studies of free-radical reactions.
    The Biochemical journal, 1988, Oct-15, Volume: 255, Issue:2

    The reactions between Trolox C, a water-soluble vitamin E analogue, and several oxidizing free radicals including the hydroxyl radical and various peroxy radicals were examined by using the pulse-radiolysis technique. The results demonstrate that Trolox C may undergo rapid one-electron-transfer reactions as well as hydrogen-transfer processes; the resulting phenoxyl radical is shown to be relatively stable, in common with the phenoxyl radical derived from vitamin E. The reactions between the Trolox C phenoxyl radical and a variety of biologically relevant reducing compounds were examined by using both pulse radiolysis and e.s.r. The results demonstrate that the Trolox C phenoxyl radical is readily repaired by ascorbate (k = 8.3 x 10(6) dm3.mol-1.s-1) and certain thiols (k less than 10(5) dm3.mol-1.s-1) but not by urate, NADH or propyl gallate. Evidence from e.s.r. studies indicates that thiol-containing compounds may also enter into similar repair reactions with the alpha-tocopherol phenoxyl radical. Kinetic evidence is presented that suggests that Trolox C may 'repair' proteins that have been oxidized by free radicals.

    Topics: Ascorbic Acid; Benzopyrans; Benzothiazoles; Binding, Competitive; Chromans; Electron Spin Resonance Spectroscopy; Electron Transport; Free Radicals; Hydrogen-Ion Concentration; Indicators and Reagents; Oxidation-Reduction; Pulse Radiolysis; Sulfhydryl Compounds; Sulfonic Acids

1988
Ozone effects on inhibitors of human neutrophil proteinases.
    Archives of biochemistry and biophysics, 1987, Feb-15, Volume: 253, Issue:1

    The effects of ozone on human alpha 1-proteinase inhibitor (A-1-PI), alpha 1-antichymotrypsin (A-1-Achy), bronchial leukocyte proteinase inhibitor (BLPI), and Eglin C were studied using in vitro exposures in phosphate-buffered solutions. Following ozone exposure, inhibitory activities against human neutrophil elastase (HNE) and/or cathepsin G (Cat G) were measured. Exposure of A-1-PI to 50 mol O3/mol protein resulted in a complete loss of HNE inhibitory activity, whereas A-1-Achy lost only 50% of its Cat G inhibitory activity and remained half active even after exposure to 250 mol of O3. At 40 mol O3/mol protein, BLPI lost 79% of its activity against HNE and 87% of its Cat G inhibitory activity. Eglin C, a leech-derived inhibitor, lost 81% of its HNE inhibitory activity and 92% of its ability to inhibit Cat G when exposed to 40 mol O3/mol. Amino acid analyses of ozone-exposed inhibitors showed destruction of Trp, Met, Tyr, and His with as little as 10 mol O3/mol protein, and higher levels of O3 resulted in more extensive oxidation of susceptible residues. The variable ozone susceptibility of the different amino acid residues in the four proteins indicated that oxidation was a function of protein structure, as well as the inherent susceptibility of particular amino acids. Exposure of A-1-PI and BLPI in the presence of the antioxidants, Trolox C (water soluble vitamin E) and ascorbic acid (vitamin C), showed that antioxidant vitamins may protect proteins from oxidative inactivation by ozone. Methionine-specific modification of BLPI reduced its HNE and Cat G inhibitory activities. Two moles of N-chlorosuccinimide per mole of BLPI methionine caused an 80% reduction in activity against Cat G, but only a 40% reduction in HNE inhibitory activity.

    Topics: alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; Amino Acids; Ascorbic Acid; Chromans; Humans; Kinetics; Neutrophils; Oxidation-Reduction; Ozone; Protease Inhibitors; Proteins; Serpins

1987
Antioxidant and co-antioxidant activity of vitamin C. The effect of vitamin C, either alone or in the presence of vitamin E or a water-soluble vitamin E analogue, upon the peroxidation of aqueous multilamellar phospholipid liposomes.
    Biochimica et biophysica acta, 1985, Jul-09, Volume: 835, Issue:2

    Thermally labile azo-initiators, dissolved in either the aqueous or lipid phase, have been used to generate peroxyl radicals at a known, steady rate in an aqueous dispersion of dilinoleoylphosphatidylcholine multilamellar liposomes at 37 degrees C in order to study the antioxidant behaviour of ascorbate itself and ascorbate in combination with either alpha-tocopherol or a water-soluble alpha-tocopherol analogue (TROLOX(-]. It is found that ascorbate is an effective inhibitor of peroxidations initiated in the aqueous phase, with each ascorbate terminating 0.6 radical chains (i.e., n = 0.6), but it is a very poor inhibitor of peroxidations initiated in the lipid phase. Peroxidations initiated in the lipid-phase in the presence of either alpha-tocopherol or TROLOX(-) indicate that ascorbate is an excellent synergist with both phenolic antioxidants (n = 0.4). In peroxidations initiated in the aqueous phase ascorbate acts as a co-antioxidant with TROLOX(-) (n = 0.7), but the interpretation of the approximately additive effect obtained in the presence of alpha-tocopherol is complicated by the fact that under the experimental conditions employed alpha-tocopherol alone does not give a distinct, measurable inhibition period. The latter problem is shown to be due to a non-uniform distribution of the water-soluble initiator within the liposome. Other examples of the complicating effects of non-uniform distributions of reactants in kinetic studies of the autoxidation of organic substrates dispersed in water are described.

    Topics: Antioxidants; Ascorbic Acid; Benzopyrans; Chromans; Kinetics; Liposomes; Models, Biological; Oxidation-Reduction; Oxygen Consumption; Phosphatidylcholines; Vitamin E

1985