ascorbic-acid and Hemolysis

There is an increased number of reports being published on rasburicase-induced methemoglobinemia recently. We aimed to identify and critically evaluate all the descriptive studies that described the rasburicase-induced methemoglobinemia, its treatment approach, and their outcomes.. PubMed, Scopus and grey literature databases were searched from inception to January 2022 using search terms "rasburicase" and "methemoglobinemia" without any language and date restriction. A bibliographic search was also done to find additional studies. Only descriptive studies on Rasburicase-induced methemoglobinemia were included for our review. Two contributors worked independently on study selection, data abstraction, and quality assessment, and any disagreements were resolved by consensus or discussion with a third reviewer.. A total of 24 reports including 27 patients (23 male, 3 female patients, and 1 study did not specify the gender of the patient) aged from 5 to 75 years were included in the review. Immediate withdrawal of the drug and administering methylene blue, ascorbic acid, blood transfusion, and supportive oxygen therapy are the cornerstone in the management of rasburicase-induced methemoglobinemia.. Rasburicase administration should be followed by careful monitoring of patients for any severe complication and treat it as early as possible appropriately. In a patient who presents with rasburicase-induced haemolysis or methemoglobinemia, it is often important to expect a diagnosis of G6PD deficiency unless otherwise confirmed and to avoid administering methylene blue, even though the patient is from a low-risk ethnicity for G6PDD.

Topics: Ascorbic Acid; Female; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Male; Methemoglobinemia; Methylene Blue

Topics: Anemia; Ascorbic Acid; Blood Coagulation Disorders; Chronic Disease; Folic Acid; Hematologic Diseases; Hemolysis; Humans; Leukocytosis; Mononuclear Phagocyte System; Serum Albumin; Stress, Physiological; Syndrome; Thrombocytosis; Zinc

Topics: Absorption; Amino Acids; Anemia, Hypochromic; Animals; Ascorbic Acid; Bile; Binding Sites; Chelating Agents; Diet; Epithelium; Erythropoiesis; Female; Gastric Juice; Gastric Mucosa; Hemolysis; Hexoses; Humans; Intestinal Absorption; Intestinal Mucosa; Iron; Pregnancy; Rats

Topics: Adult; Anemia, Hemolytic; Anemia, Macrocytic; Ascorbic Acid; Ascorbic Acid Deficiency; Blood Cell Count; Bone Marrow Cells; Erythrocytes; Erythropoiesis; FIGLU Test; Folic Acid; Folic Acid Deficiency; Hemolysis; Hemorrhagic Disorders; Humans; Iron; Leukocytes; Liver; Reticulocytes; Scurvy; Tissue Extracts; Vitamin B 12; Vitamin B 12 Deficiency

Topics: Ascorbic Acid; Carbohydrate Metabolism; Cysteine; Erythrocyte Aging; Erythrocytes; Glucosephosphate Dehydrogenase; Hemolysis; Humans; NADP; Pentosephosphates; Pyruvates

Sepsis is a health problem of global importance; treatments focus on controlling infection and supporting failing organs. Recent clinical research suggests that intravenous vitamin C may decrease mortality in sepsis. We have designed a randomized controlled trial (RCT) to ascertain the effect of vitamin C on the composite endpoint of death or persistent organ dysfunction at 28 days in patients with sepsis.. LOVIT (Lessening Organ dysfunction with VITamin C) is a multicenter, parallel-group, blinded (participants, clinicians, study personnel, Steering Committee members, data analysts), superiority RCT (minimum n = 800). Eligible patients have sepsis as the diagnosis for admission to the intensive care unit (ICU) and are receiving vasopressors. Those admitted to the ICU for more than 24 h are excluded. Eligible patients are randomized to high-dose intravenous vitamin C (50 mg/kg every 6 h for 96 h) or placebo. The primary outcome is a composite of death or persistent organ dysfunction (need for vasopressors, invasive mechanical ventilation, or new and persisting renal replacement therapy) at day 28. Secondary outcomes include persistent organ dysfunction-free days to day 28, mortality and health-related quality of life at 6 months, biomarkers of dysoxia, inflammation, infection, endothelial function, and adverse effects (hemolysis, acute kidney injury, and hypoglycemia). Six subgroup analyses are planned.. This RCT will provide evidence of the effect of high-dose intravenous vitamin C on patient-important outcomes in patients with sepsis.. clinicaltrials.gov, NCT03680274, first posted 21 September 2018.

Topics: Acute Kidney Injury; Administration, Intravenous; Adult; Antioxidants; Ascorbic Acid; Clinical Trials, Phase III as Topic; Dose-Response Relationship, Drug; Drug Therapy, Combination; Female; Hemolysis; Hospital Mortality; Humans; Hypoglycemia; Intensive Care Units; Male; Middle Aged; Multiple Organ Failure; Quality of Life; Sepsis; Treatment Outcome; Vasoconstrictor Agents

To compare the effects of lotus root and cucumber on antioxidant function in aged subjects.. Pilot comparative study.. Research setting with vegetable intervention.. Healthy aged subjects over the age of sixty.. 30-day supplementation of lotus root or cucumber powder.. Plasma value of ferric reducing antioxidant power assay, activity of antioxidant enzymes, contents of some antioxidants, oxidation products, hemolysis, blood mononuclear cell DNA damage and urinary excretion of 8-hydroxy-2'-deoxyguanosine were measured before and after the intervention.. Plasma glutathione peroxidase activity, contents of vitamin C, total phenolics were significantly increased, while plasma uric acid content significantly decreased in both groups at the end of the intervention. Meanwhile, hemolysis was significantly reduced in both groups and DNA injury rate of blood mononuclear cells in lotus root group and the ratio of comet tail length to total length in cucumber group were also declined significantly post-intervention. However, plasma value of ferric reducing antioxidant power assay, contents of reduced glutathione, vitamin E, malondialdehyde, oxidized low density lipoprotein, carbonyls and activity of superoxide dismutase and catalase were not changed significantly in both group after the intervention.. These results suggest that lotus root and cucumber are not remarkably different in improving antioxidant function in aged subjects, though they are significantly different in antioxidant capacity in vitro. The benefits observed in this study may come from the additive or synergistic combinations of antioxidants contained in vegetables.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Aged; Antioxidants; Ascorbic Acid; Catalase; Comet Assay; Cucumis sativus; Deoxyguanosine; Dietary Supplements; DNA Damage; Female; Glutathione; Glutathione Peroxidase; Healthy Volunteers; Hemolysis; Humans; Leukocytes, Mononuclear; Lipoproteins, LDL; Lotus; Male; Malondialdehyde; Middle Aged; Oxidation-Reduction; Phenols; Pilot Projects; Superoxide Dismutase; Uric Acid; Vegetables; Vitamin E

Changes that occur to red blood cells (RBCs) during routine blood bank storage include decreased deformability, increased haemolysis and oxidative damage. Oxidative injury to the RBC membrane and haemoglobin can affect changes in shape and deformability. Ascorbic acid (AA) is an antioxidant that maintains haemoglobin in a reduced state and minimises RBC oxidative injury. We hypothesised that AA would improve membrane fragility and decrease haemolysis during storage.. Whole blood derived, AS-5 preserved, pre-storage leucoreduced RBC units were exposed to either AA or saline control solutions. Several rheological and biochemical parameters were measured serially during storage, including RBC membrane mechanical fragility, percent haemolysis and methaemoglobin levels.. AA exposure significantly reduced mechanical fragility and haemolysis over the entire storage period. The highest two concentrations of AA affected the greatest reductions in mechanical fragility and percent haemolysis. Addition of AA to the RBCs did not significantly alter their biochemical parameters compared to control RBCs incubated with saline.. AA reduced RBC membrane fragility and decreased haemolysis during storage without adversely affecting other RBC biochemical parameters. The clinical significance of these findings needs to be determined.

Topics: Adult; Antioxidants; Ascorbic Acid; Blood Preservation; Erythrocyte Deformability; Erythrocyte Membrane; Female; Hemoglobins; Hemolysis; Humans; Male; Middle Aged; Oxidation-Reduction

Erythrocytes from sickle cell anaemia (SCA) patients continuously produce larger amounts of pro-oxidants than normal cells. Oxidative stress seems to primarily affect the membrane and results in haemolysis. The use of antioxidants in vitro reduces the generation of pro-oxidants. To evaluate the impact of vitamins C (VitC) and E (VitE) supplementation in SCA patients, patients over 18 years were randomly assigned to receive VitC 1400 mg + VitE 800 mg per day or placebo orally for 180 d. Eighty-three patients were enrolled (44 vitamins, 39 placebo), median age 27 (18-68) years, 64% female. There were no significant differences between the two groups regarding clinical complications or baseline laboratorial tests. Sixty percent of the patients were VitC deficient, 70% were VitE deficient. Supplementation significantly increased serum VitC and E. However, no significant changes in haemoglobin levels were observed, and, unexpectedly, there was a significant increase in haemolytic markers with vitamin supplementation. In conclusion, VitC + VitE supplementation did not improve anaemia and, surprisingly, increased markers of haemolysis in patients with SCA and S-β(0) -thalassaemia. The exact mechanisms to explain this findings and their clinical significance remain to be determined.

Topics: Adolescent; Adult; Aged; Anemia, Sickle Cell; Antioxidants; Ascorbic Acid; Biomarkers; Dietary Supplements; Double-Blind Method; Drug Utilization; Female; Hemolysis; Hospitalization; Humans; Inflammation; Male; Middle Aged; Oxidative Stress; Quality of Life; Reactive Oxygen Species; Sickle Cell Trait; Thalassemia; Vitamin E; Young Adult

Chronic hemodialysis (HD) patients manifest anemia and atherosclerosis with associated oxidative stress. We explored whether intravenous infusion of vitamin C (VC) and/or use of vitamin E (VE)-coated dialysis membrane could palliate HD-evoked oxidative stress. Eighty patients undergoing chronic HD were enrolled and randomly assigned into four groups: HD with intravenous VC (n=20), HD with VE-coated dialyzer (n=20), HD with both (n=20), and HD with neither (n=20). We evaluated oxidative stress in blood and plasma, erythrocyte methemoglobin/ferricyanide reductase (red blood cells (RBC)-MFR) activity, plasma methemoglobin, and pro-inflammatory cytokines in these patients. All patients showed marked increases (14-fold) in blood reactive oxygen species (ROS) after HD. The types of ROS were mostly hydrogen peroxide, and in lesser amounts, O2*- and HOCl. HD resulted in decreased plasma VC, total antioxidant status, and RBC-MFR activity and increased plasma and erythrocyte levels of phosphatidylcholine hydroperoxide (PCOOH) and methemoglobin. Intravenous VC significantly palliated HD-induced oxidative stress, plasma and RBC levels of PCOOH, and plasma methemoglobin levels and preserved RBC-MFR activity. The VE-coated dialyzer effectively prevented RBCs from oxidative stress, although it showed a partial effect on the reduction of total ROS activity in whole blood. In conclusion, intravenous VC plus a VE-coated dialyzer is effective in palliating HD-evoked oxidative stress, as indicated by hemolysis and lipid peroxidation, and by overexpression of proinflammation cytokines in HD patients. Using VE-coated dialyzer per se is, however, effective in reducing lipid peroxidation and oxidative damage to RBCs.

Topics: Antioxidants; Ascorbic Acid; Cytokines; Erythropoietin; Female; Hemolysis; Humans; Hydrogen Peroxide; Infusions, Intravenous; Kidney Failure, Chronic; Lipid Peroxidation; Male; Membranes, Artificial; Methemoglobin; NADH, NADPH Oxidoreductases; Oxalates; Oxidative Stress; Phosphatidylcholines; Reactive Oxygen Species; Renal Dialysis; Spectrophotometry, Atomic; Vitamin E

alpha-Tocopherol is a potent antioxidant that effectively protects biological membranes against oxidative injury through coordination with ascorbic acid. Because propofol has a phenolic structure similar to that of alpha-tocopherol, this intravenous anesthetic may also have similar antioxidant activity. To test this hypothesis, the effect of propofol on oxidative injury of human erythrocytes was examined. Propofol inhibited oxidative hemolysis and cis-parinaric acid oxidation in erythrocyte membranes (ED(50) = 6 microM). Although ascorbic acid alone has no appreciable effect, the protective effect of propofol was enhanced by ascorbic acid. An electron spin resonance (ESR) study showed that propofol-derived radicals (g = 2.005) were continuously generated during the oxidation of erythrocyte membranes by an ascorbic acid-inhibitable mechanism. These and other results suggest that propofol interacts with ascorbic acid, thereby exhibiting potent antioxidant activity in and around membranes as does alpha-tocopherol. Kinetic analysis revealed that propofol increased the membrane fluidity of erythrocytes, thereby increasing their resistance to physical and hemodynamic stress. Further, a greater preservation of red blood cell counts was seen after surgery with propofol compared with conventional sevoflurane anesthesia. Thus, propofol may protect erythrocytes against both oxidative and physical stress, indicating its potential as an efficient and safe antioxidant.

Topics: alpha-Tocopherol; Antioxidants; Ascorbic Acid; Drug Evaluation, Preclinical; Drug Synergism; Electron Spin Resonance Spectroscopy; Erythrocyte Count; Erythrocyte Membrane; Fatty Acids, Unsaturated; Free Radical Scavengers; Free Radicals; Gastrectomy; Hemolysis; Humans; Intraoperative Care; Middle Aged; Osmotic Fragility; Oxidative Stress; Propofol; Stress, Mechanical

We tested the hypothesis that vitamin C supplementation of premature neonates is associated with hemolysis.. A double-blind, randomized, controlled trial of vitamin C supplementation (50 mg/day) was undertaken in premature neonates (birth weight, 1000 to 1500 gm). Infants were randomly assigned to receive vitamin C (Ce-Vi-Sol) (n = 32) or placebo (n = 24) for 14 days. Twenty-three subjects per group were required to detect a difference of 1 SD in corrected carboxyhemoglobin values (alpha = 0.05, beta = 0.10).. Day 14 vitamin C levels were lower in control subjects than in supplemented neonates (62 +/- 24 vs 125 +/- 62 micromol/L, p = 0.005). There was no difference in corrected blood carboxyhemoglogin concentrations (0.72 +/- 0.44 vs 0.72 +/- 0.23%; p = 0.95), other parameters of hemolysis, weight gain, blood sampled, presumed septic episodes, necrotizing enterocolitis, feeding intolerance, or transfusion. On day 14, bilirubin values were higher in control subjects than in the supplemented group (77 +/- 37 vs 55 +/- 33 micromol/L; p = 0.04). When a distant outlier in the nonsupplemented group was excluded (163 micromol/L), statistical significance was lost (73 +/- 32 vs 55 +/- 33 micromol/L; p = 0.09).. Oral supplementation of premature infants with vitamin C is not associated with evidence of increased erythrocyte destruction, hyperbilirubinemia, or other morbidity.

Topics: Anemia, Hemolytic; Ascorbic Acid; Double-Blind Method; Female; Hemolysis; Humans; Hyperbilirubinemia; Infant, Newborn; Infant, Premature; Infant, Premature, Diseases; Male

The aim of this study was to examine whether extreme endurance stress of trained athletes can influence lipid peroxidation and muscle enzymes. A randomized and placebo-controlled study was carried out on 24 trained long-distance runners who were substituted with alpha-tocopherol (400 I.U. d-1) and ascorbic acid (200 mg d-1) during 4.5 weeks prior to a marathon race. The serum concentrations of retinol, ascorbic acid, beta-carotene, alpha-tocopherol, malondialdehyde (TBARS) and uric acid as well as glutathione peroxidase (GSH Px) and catalase were measured 4.5 weeks before (A), immediately before (B), immediately after (C) and 24 h after (D) the course. After competition (C) TBARS serum concentrations of the athletes (n = 22) decreased in both groups (P < 0.0001). The ascorbic acid serum concentration increased significantly in the supplemented group from (A) to (B) (P < 0.01), from (B) to (C) (P < 0.001) and in the placebo group a significant increase from (B) to (C) (P < 0.01) was observed. The alpha-tocopherol serum concentration increased significantly in the supplemented group from (A) to (B) (P < 0.001) and from (B) to (C) (P < 0.05). The enzymes glutathione peroxidase (GSH Px) and catalase measured in erythrocytes as well as the serum selenium levels did not show significant differences at any time. A significant increase of CK concentration was observed from (C) to (D) in the supplemented group (P < 0.01) and in the placebo group (P < 0.001). The increase of CK serum concentration is remarkably lower in the supplemented group compared with the placebo group (P < 0.01). It is concluded that endurance training coupled with antioxidant vitamin supplementation reduces blood CK increase under exercise stress.

Topics: Adult; Antioxidants; Ascorbic Acid; Blood Volume; Creatine Kinase; Exercise Test; Hemolysis; Humans; Lipid Peroxidation; Male; Middle Aged; Oxidative Stress; Physical Endurance; Physical Exertion; Thiobarbituric Acid Reactive Substances; Vitamin E

Due to its advantageous antioxidant phytochemical components,

Topics: Animals; Antioxidants; Ascorbic Acid; Catechin; Flavonoids; Hemolysis; Humans; Oils, Volatile; Phytochemicals; Plant Extracts; Plant Leaves; Psidium

Background In this study, the hematological and antioxidant potential as well as the osmotic fragility effects of a Nigerian polyherbal formulation were evaluated. Materials and methods A total of 40 fats were divided into four groups of 10 rats each. Group 1 served as the control group, and the rest were assigned increasing daily oral administration of the extract for 28 days. At the end of treatment, blood was collected for hematological and osmotic fragility studies. The free radical scavenging effect of the extract was investigated via different in vitro models as well. Results Results showed that the nitric oxide scavenging and 2,2-diphenyl-1-picrylhydrazyl (DPPH) activities of the extract were significant (p < 0.05) and compared favorably with that of vitamin C. At 200 and 400 μg/mL, the nitric oxide scavenging activities for Ajumbise Polyherbal Extract (APE) were 60.71 ± 0.25% and 59.49 ± 0.98%, respectively, whereas for the same concentrations of vitamin C, 74.60 ± 0.25% and 85.24 ± 0.14 scavenging activities were obtained. The (DPPH) activity at 100 μg/mL was 81.24 ± 0.02% for the extract and 96.22 ± 0.18% for vitamin C. However, at all concentrations, the extract had significantly lower Ferric Reducing Antioxidant Power (FRAP) activity than vitamin C. Red blood cell counts (RBCC), hemoglobin and packed cell volume values (PCV) were significantly lowered only in groups treated with 400 and 800 mg/kg of the extract (p < 0.05), whereas other RBCC parameters and white blood cell counts (WBCC) were not significantly affected (p < 0.05). Platelet (PLT) count was also significantly lowered in all extract-treated groups. The extract also significantly reduced RBCC percentage hemolysis (p < 0.05). Conclusions Ajumbise polyherbal may be free of hematoxicity and may improve the integrity of the RBC membrane due to its appreciable antioxidant activity.

Topics: Animals; Antioxidants; Ascorbic Acid; Biphenyl Compounds; Blood Platelets; Erythrocytes; Flavonoids; Hematology; Hemolysis; Leukocytes; Nigeria; Nitric Oxide; Osmotic Fragility; Phenols; Phytotherapy; Picrates; Plant Extracts; Plants, Medicinal; Rats

Diabetes mellitus (DM) is a major health problem with an increasing global prevalence. It is usually associated with an imbalance between pro-oxidant mechanisms and antioxidant defenses, contributing to oxidative-stress, and this leads to an increased susceptibility to endothelial dysfunction, atherosclerosis, insulin-resistance and impaired-pancreatic β-cell function.. We have assessed the Prooxidant-antioxidant balance (PAB) and anti-hemolytic effect of Thuja orientalis L using the PAB assay and the analysis of hematological markers.. The antioxidant and anti-hemolytic activity of Thuja orientalis was evaluated using the PAB assay and the inhibition of RBC hemolysis using the hydrogen peroxide hemolysis test. The percentage of anti-hemolysis was calculated from the ratio of the measurements (A-B)/B×100.. Our results showed that the antioxidant effect of Thuja orientalis L. was greater in water than in ethyl-acetate, ethanol and methanol extract, respectively. We also observed its anti-hemolytic effect, which was higher in water, than in ethyl-acetate, methanol and ethanol extract, respectively. In particular our data showed that the H2O2-induced RBC hemolysis was inhibited in a dose-dependent manner.. we demonstrated the antioxidant and anti-hemolytic effect of Thuja orientalis L. extracts in human serum and RBC, showing its potential property of reducing free radicals supporting further investigations to assess its functional role in larger samples size and in vivo models, as a potential antioxidant agent.

Topics: Antioxidants; Ascorbic Acid; Diabetes Mellitus; Hemolysis; Humans; Oxidants; Plant Extracts; Thuja

Topics: Animals; Anti-Anxiety Agents; Antioxidants; Artemia; Ascorbic Acid; Behavior, Animal; Biomarkers; Catalase; Coumaric Acids; Diazepam; Erythrocytes; gamma-Aminobutyric Acid; Glutathione; Glutathione Peroxidase; Hemolysis; Hippocampus; Lipid Peroxidation; Male; Mice; Nitrites; Oxidative Stress; Superoxide Dismutase; Thiobarbituric Acid Reactive Substances; Toxicity Tests

Topics: Animals; Ascorbic Acid; Biological Transport; Cell Membrane; Dehydroascorbic Acid; Erythrocytes; Gene Knockout Techniques; Glucose; Hemolysis; Mice; Mice, Knockout; Oxidation-Reduction; Oxidoreductases; Phenotype; Sugar Acids

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

In experiments in vivo we studied the effect of chronic iron-contained drug (Urfuhem) supplemention on the level of hemoglobin (Hb) in the blood of aging rats. To establish the biochemical mechanisms of drug action it were determined the parameters of oxidative/nitrosative stress and the hydrogei sulfide level in plasma and erythrocytes, the level of non-heme iron in plasma and erythrocytes sensitivity to acid hemolysis. It was found that in aging rats the drug significantly increases the Hb content of red blood cells and reduces its resistance to acid hemolysis. After the drug supplemention the rate of superoxide anion-radical (*0(2)(-)) generation in erythrocytes and stable hydrogen peroxide (H(2)0(2)) content both in plasma and erythrocytes, were down-regulated. The drug did not reduce the high levels of generation of the hydroxyl radical (*OH) and high levels of excess NO de novo synthesis by iNOS in erythrocytes but reduced the pools of nitrate anion(NO(3)(-))a its reutilization for NO synthesis. After thei drug supplemention the rate of constitutine NO synthesis by cNOS in aging rats plasma was up-regulated perhaps by cNOS coupling. The results indicate that the reason for increasing the permeability of the proton (H*) in red blood cells that causes the acid hemolysis in aging fats after the drug supplemention can be change in the balance of levels of oxidative and nitrosative stress in red blood cells in favor of the latter, and that toxic, OH generation is not at the expense of the classical Fenton reaction in the presence of iron ions (Fe(2+)); but due to the formation and decomposition of peroxynitrie (ON0O(-)).

Topics: Aging; Animals; Animals, Outbred Strains; Ascorbic Acid; Erythrocytes; Hemoglobins; Hemolysis; Hydrogen Peroxide; Hydrogen-Ion Concentration; Hydroxyl Radical; Iron; Lipid Peroxidation; Primary Cell Culture; Rats; Superoxides

A 15-year-old boy presented to emergency services with accidental naphthalene ball ingestion. Following consumption he developed methaemoglobinaemia, massive intravascular haemolysis and acute kidney injury. He had no history suggestive of congenital haemoglobin M disease. Development of severe methaemoglobinaemia and intravascular haemolysis is quite unusual after consumption of a single ball of naphthalene. The patient was managed with ascorbic acid and intravenous N-acetyl cysteine. He also required haemodialysis for acute kidney injury that developed secondary to pigment nephropathy.

Topics: Acute Kidney Injury; Adolescent; Antioxidants; Ascorbic Acid; Cysteine; Hemolysis; Humans; Male; Methemoglobinemia; Naphthalenes; Renal Dialysis

The discovery that oxidized vitamin C, dehydroascorbate (DHA), can induce oxidative stress and cell death in cancer cells has rekindled interest in the use of high dose vitamin C (VC) as a cancer therapy. However, high dose VC has shown limited efficacy in clinical trials, possibly due to the decreased bioavailability of oral VC. Because human erythrocytes express high levels of Glut1, take up DHA, and reduce it to VC, we tested how erythrocytes might impact high dose VC therapies. Cancer cells are protected from VC-mediated cell death when co-cultured with physiologically relevant numbers of erythrocytes. Pharmacological doses of VC induce oxidative stress, GSH depletion, and increased glucose flux through the oxidative pentose phosphate pathway (PPP) in erythrocytes. Incubation of erythrocytes with VC induced hemolysis, which was exacerbated in erythrocytes from glucose-6-phosphate dehydrogenase (G6PD) patients and rescued by antioxidants. Thus, erythrocytes protect cancer cells from VC-induced oxidative stress and undergo hemolysis in vitro, despite activation of the PPP. These results have implications on the use of high dose VC in ongoing clinical trials and highlight the importance of the PPP in the response to oxidative stress.

Topics: Ascorbic Acid; Cell Line, Tumor; Dehydroascorbic Acid; Erythrocytes; Glucose; Glucose Transporter Type 1; Glucosephosphate Dehydrogenase; Glutathione; Hemolysis; Humans; Neoplasms; Oxidation-Reduction; Oxidative Stress; Pentose Phosphate Pathway

There is an urgent need to develop blood-contacting biomaterials with long-term anti-hemolytic capability. To obtain such biomaterials, we coaxially electrospin [ascorbic acid (AA) and lecithin]/poly (ethylene oxide) (PEO) core-shell nanofibers onto the surface of styrene-b-(ethylene-co-butylene)-b-styrene elastomer (SEBS) that has been grafted with poly (ethylene glycol) (PEG) chains. Our strategy is based on that the grafted layers of PEG render the surface hydrophilic to reduce the mechanical injure to red blood cells (RBCs) while the AA and lecithin released from nanofibers on blood-contacting surface can actively interact with RBCs to decrease the oxidative damage to RBCs. We demonstrate that (AA and lecithin)/PEO core-shell structured nanofibers have been fabricated on the PEG grafted surface. The binary release of AA and lecithin in the distilled water is in a controlled manner and lasts for almost 5 days; during RBCs preservation, AA acts as an antioxidant and lecithin as a lipid supplier to the membrane of erythrocytes, resulting in low mechanical fragility and hemolysis of RBCs, as well as high deformability of stored RBCs. Our work thus makes a new approach to fabricate blood-contacting biomaterials with the capability of long-term anti-hemolysis.

Topics: Animals; Antioxidants; Ascorbic Acid; Biocompatible Materials; Cells, Cultured; Elastomers; Electrochemical Techniques; Erythrocyte Deformability; Erythrocytes; Hemolysis; Hydrophobic and Hydrophilic Interactions; Lecithins; Microscopy, Electron, Scanning; Nanofibers; Polyethylene Glycols; Rabbits; Styrenes; Surface Properties

Potential risk of high-dose vitamin C consumption is often ignored. Recently, gram-dose vitamin C is being intravenously injected for the treatment of cancer, which can expose circulating blood cells to extremely high concentrations of vitamin C. As well as platelets, red blood cells (RBCs) can actively participate in thrombosis through procoagulant activation. Here, we examined the procoagulant and prothrombotic risks associated with the intravenous injection of gram-dose vitamin C. Vitamin C (0.5-5 mM) increased procoagulant activity of freshly isolated human RBCs via the externalization of phosphatidylserine (PS) to outer cellular membrane and the formation of PS-bearing microvesicles. PS exposure was induced by the dysregulation of key enzymes for the maintenance of membrane phospholipid asymmetry, which was from vitamin C-induced oxidative stress, and resultant disruption of calcium and thiol homeostasis. Indeed, the intravenous injection of vitamin C (0.5-1.0 g/kg) in rats in vivo significantly increased thrombosis. Notably, the prothrombotic effects of vitamin C were more prominent in RBCs isolated from cancer patients, who are at increased risks of thrombotic events. Vitamin C-induced procoagulant and prothrombotic activation of RBCs, and increased thrombosis in vivo. RBCs from cancer patients exhibited increased sensitivity to the prothrombotic effects of vitamin C, reflecting that intravenous gram-dose vitamin C therapy needs to be carefully revisited.

Topics: Adenosine Triphosphate; Animals; Ascorbic Acid; Blood Coagulation; Calcium; Erythrocytes; Flow Cytometry; Glutathione; Hemolysis; Humans; Injections, Intravenous; Leukemia; Male; Microscopy, Electron, Scanning; Neoplasms; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Thrombosis; Vitamins

To assess changes in biochemical and biophysical properties of canine RBCs during cold (1° to 6°C) storage in a licensed RBC additive solution (the RBC preservation solution designated AS-1) supplemented with ascorbic acid.. Blood samples from 7 neutered male Greyhounds; all dogs had negative results when tested for dog erythrocyte antigen 1.1.. Blood was collected into citrate-phosphate-dextrose and stored in AS-1. Stored RBCs were supplemented with 7.1mM ascorbic acid or with saline (0.9% NaCl) solution (control samples). Several biochemical and biophysical properties of RBCs were measured, including percentage hemolysis, oxygen-hemoglobin equilibrium, and the kinetic rate constants for O2 dissociation, carbon monoxide association, and nitric oxide dioxygenation.. Greyhound RBCs stored in AS-1 supplemented with ascorbic acid did not have significantly decreased hemolysis, compared with results for the control samples, during the storage period.. In this study, ascorbic acid did not reduce hemolysis during storage. Several changes in stored canine RBCs were identified as part of the hypothermic storage lesion.

Topics: Animals; Ascorbic Acid; Blood Preservation; Cold Temperature; Dogs; Erythrocytes; Hemolysis; Male; Sodium Chloride; Time Factors

Five 4-arylcoumarins (1c-g) and twelve 3,4-dihydro-4-arylcoumarins (2a-l) were synthesized and tested for antioxidant activity, antitumor activity, toxicity and structure-activity relationships analysis. 4-Arylcoumarins and 3,4-dihydro-4-arylcoumarins that possess two hydroxyl groups in ortho position, such as 1d, 1f, 2a, 2f, 2g and 2h had stronger radical scavenging properties than that of vitamin C (Vit C) in ABTS(+) assay. Kinetic traces of scavenging ABTS(+) and DPPH radicals showed that all the reaction could reached endpoint in 1 min, which was similar with Vit C. 4-Arylcoumarins with 3'-hydroxyl-4'-methylphenyl structural show more efficient NO radical scavenging activity. Three compounds 2e, 1f and 2a, in particular had superior EC50 for NO scavenging than did Vit C. MTT assay indicated that one compound in particular had a potential antitumor effect, inhibiting proliferation of BGC-823 cells and almost completely killing them at a concentration 62.5 mg/L. With same concentration 100 μg/mL, hemolytic analysis in rabbit red blood cells showed that only two compounds had hemolytic activity with a little more than 5% hemolysis. Injection and oral toxicity tests on Galleria mellonella larvae showed that none of the tested 4-arylcoumarins significantly affected their appetite, viability and mortality.

Topics: Administration, Oral; Animals; Antineoplastic Agents; Antioxidants; Ascorbic Acid; Cell Line, Tumor; Coumarins; Drug Evaluation, Preclinical; Erythrocytes; Free Radical Scavengers; Hemolysis; Humans; Molecular Structure; Moths; Rabbits; Structure-Activity Relationship; Toxicity Tests

Hawk tea (Litsea coreana var. lanuginose) is a very popular herbal tea in the southwest of China. According to the maturity degree of raw materials, Hawk tea can usually be divided into three types: Hawk bud tea (HB), Hawk primary leaf tea (HP), and Hawk mature leaf tea (HM). In this study, some of the bioactive constituents and antioxidant properties of the three kinds of Hawk tea infusions were comparatively investigated. The results showed that the contents of total flavonoids, vitamin C, and carbohydrates in Hawk bud tea infusion (HBI) were higher than those in Hawk primary leaf tea infusion (HPI) and Hawk mature leaf tea infusion (HMI). HPI had higher contents of total polyphenols and exhibited better DPPH radical scavenging activity and ferric reducing activity power. HBI could provide more effective protection against erythrocyte hemolysis. As age is going from bud to mature leaf, the ability to inhibit the formation of low density lipoprotein (LDL) conjugated diene and the loss of tryptophan fluorescence decreased. The bioactive constituents and antioxidant activities of Hawk tea infusions were significantly affected by the maturity degree of the raw material.

Topics: Ascorbic Acid; Carbohydrates; Cells, Cultured; Flavonoids; Free Radical Scavengers; Hemolysis; Humans; Litsea; Oxidation-Reduction; Plant Extracts

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

Topics: Acute Disease; Ascorbic Acid; Dose-Response Relationship, Drug; Erythrocytes; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Infant; Male; Tachycardia

Amphotericin B (AmB) remains the antifungal polyene of choice in deep fungal infections, but its high toxicity to mammalian cells limits its use. This toxicity is partly due to lipid peroxidation exerted by amphotericin B in cell membranes. The work we have undertaken focused on the one part the evaluation of the efficacy of amphotericin B in the presence of some antioxidants vitamins (vitamin C "ascorbic acid" and vitamin E "α-tocopherol") against the yeast Candida albicans ATCC 10231. Secondly, we have tested the cytotoxicity of these formulations on human red blood cells. The results showed a significant improvement in the efficiency of our formulations tested from 7% to 12% compared with amphotericin B alone at therapeutic concentrations. Furthermore, the addition of vitamin C and vitamin E protects human red blood cells against the cytotoxicity induced by amphotericin B with 17%. This is due may be to the antioxidant power of vitamins which confer protection against the autoxidation of the molecule of amphotericin B. On the other hand, it is noticed that the yeast regrows after 24h whatever in complex with vitamin C or vitamin E of the stock solution. On completion of this study, the incorporation of antioxidant vitamins that we propose to the reaction medium of antifungal improved the therapeutic index of amphotericin B.

Topics: alpha-Tocopherol; Amphotericin B; Ascorbic Acid; Candida albicans; Drug Interactions; Erythrocytes; Hemolysis; Humans; Microbial Sensitivity Tests; Oxidation-Reduction; Toxicity Tests

Reactive oxygen species (ROS) can lead to haemolysis and eventually to diseases such as thalassemia and sickle cell anaemia. Their action can be counteracted by the antihaemolytic activity of therapeutic agents. The aim of our study was to identify plants that most efficiently counteract ROS-caused haemolysis. From ten plants known for their antioxidant activity (Orobanche orientalis G. Beck, Cucumis melo L., Albizzia julibrissin Durazz, Galium verum L., Scutellaria tournefortii Benth, Crocus caspius Fischer & Meyer, Sambucus ebulus L., Danae racemosa L., Rubus fruticsos L., and Artemisia absinthium L.) we prepared 30 extracts using three extraction methods (percolation, Soxhlet, and ultrasound-assisted extraction) to see whether the extraction method affects antihaemolytic efficiency, and one extraction method (polyphenol extraction) to see how much of this action is phenol-related. Extract antihaemolytic activity was determined in mice red blood cells and compared to that of vitamin C as a known antioxidant. Nine of our extracts were more potent than vitamin C, of which G. verum (aerial parts/percolation) and S. tournefortii (aerial parts/polyphenol) extracts were the most potent, with an IC50 of 1.32 and 2.08 μg mL⁻¹, respectively. Haemolysis inhibition depended on extract concentration and the method of extraction. These plants could provide accessible sources of natural antioxidants to the pharmaceutical industry.

Topics: Animals; Antioxidants; Ascorbic Acid; Erythrocytes; Hemolysis; Hemolytic Agents; Iran; Male; Mice; Plant Components, Aerial; Plant Extracts; Reactive Oxygen Species

A series of novel N-(4-aryl-1,3-thiazol-2-yl)-2-(2,4-dioxo-1,3-thiazolidin-5-yl)acetamides (4a-k) and N-(1,3-benzothiazol-2-yl)-2-(2,4-dioxo-1,3-thiazolidin-5-yl)acetamide derivatives (4l-o) are synthesized and evaluated for their anti-inflammatory and antioxidant activity (DPPH radical scavenging, superoxide anion scavenging, lipid peroxide inhibition, erythrocyte hemolytic inhibition). Compounds 4k and 4l have exhibited good antioxidant activity in four assays, while compounds 4c, 4d, 4m, 4n and 4o have shown good DPPH radical scavenging efficacy. Compounds 4a, 4h, 4i, 4k, 4m and 4n have possessed excellent anti-inflammatory activity. N-[4-(o-methoxyphenyl)-1,3-thiazol-2-yl]-2-(2,4-dioxo-1,3-thiazolidin-5-yl)acetamide (4k) and N-(6-nitro-/methoxy-1,3-benzothiazol-2-yl)-2-(2,4-dioxo-1,3-thiazolidin-5-yl)acetamide (4m and 4n) have exhibited both antioxidant and anti-inflammatory activities.

Topics: Acetamides; Anti-Inflammatory Agents, Non-Steroidal; Cell Line; Drug Design; Erythrocytes; Free Radical Scavengers; Hemolysis; Humans; Inhibitory Concentration 50

A series of polysulfones of new structural types on the basis of azanorbornenes, 2,2-diallyl-1,1,3,3-tetraethylguanidiniumchloride and tris(diethylamino)diallylaminophosphonium salts were obtained by free radical polymerization reaction. Their antioxidant properties in lipid peroxidation induced by iron/ascorbate and in erythrocyte hemolysis by H2O2 were evaluated. The polysulfone of 2,2-diallyl-1,1,3,3-tetraethylguanidinium chloride showed protection of erythrocytes against hydrogen peroxide and thermal shock, protected epithelial cells against UV irradiation, was not toxic for erythrocytes, epithelial cells SPEV and 3T3 fibroblasts. The polysulfone of N-benzyl-2-azanorbornene-5 inhibited lipid peroxidation in liver homogenate, did not exhibit cytotoxicity in SPEV and 3T3 cells, protected these against UV irradiation. Both compounds had low bioavailability and may be recommended for further investigations as excipients to be externally applied.

Topics: 3T3 Cells; Animals; Antioxidants; Ascorbic Acid; Cell Line; Cell Survival; Dose-Response Relationship, Drug; Epithelial Cells; Guanidine; Hemolysis; Hot Temperature; Hydrogen Peroxide; Iron; Lipid Peroxidation; Male; Mice; Models, Chemical; Molecular Structure; Oxidants; Oxidative Stress; Polymers; Sulfones; Swine; Ultraviolet Rays

In this study, the acetylcholinesterase inhibition and in vitro and in vivo antioxidant activities of Ganoderma lucidum grown on germinated brown rice (GLBR) were evaluated. In antioxidant assays in vitro, GLBR was found to have strong metal chelating activity, DPPH, ABTS, hydroxyl and superoxide radical scavenging activity. Cell-based antioxidant methods were used, including lipid peroxidation on brain homogenate and AAPH-induced erythrocyte haemolysis. In antioxidant assays in vivo, mice were administered with GLBR and this significantly enhanced the activities of antioxidant enzymes in the mice sera, livers and brains. The amount of total phenolic and flavonoid compounds were 43.14 mg GAE/g and 13.36 mg CE/g dry mass, respectively. GLBR also exhibited acetylcholinesterase inhibitory activity. In addition, HPLC analyses of GLBR extract revealed the presence of different phenolic compounds. These findings demonstrate the remarkable potential of GLBR extract as valuable source of antioxidants which exhibit interesting acetylcholinesterase inhibitory activity.

Topics: Acetylcholinesterase; Animals; Antioxidants; Ascorbic Acid; beta Carotene; Carotenoids; Catalase; Chelating Agents; Cholinesterase Inhibitors; Erythrocytes; Female; Flavonoids; Free Radical Scavengers; Glutathione; Hemolysis; Lipid Peroxidation; Lycopene; Mice; Oryza; Oxidation-Reduction; Phenols; Plant Extracts; Reishi; Superoxide Dismutase

Topics: Anemia, Hemolytic; Antioxidants; Ascorbic Acid; Cyclophosphamide; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Male; Methemoglobinemia; Middle Aged; Multiple Myeloma; Myeloablative Agonists; Treatment Outcome; Tumor Lysis Syndrome; Urate Oxidase

Aging is characterized by an impaired capacity to maintain the redox balance both in physiological and pathological situations associated with an increased production of reactive oxygen species. Since the extent of this phenomenon may be influenced by an antioxidants-rich diet, we investigated the effect of supplementation with fresh red orange juice (ROJ) on biochemical and cellular biomarkers of oxidative stress in healthy, trained elderly women after a single bout of exhaustive exercise (EE). To this purpose, a sample of 22 females, 15 (69.0 ± 5.1 years) taking the ROJ supplementation and 7 (68.1 ± 2.7 years) as Control group, was constituted. Blood samples were collected immediately before, 30 minutes, and 24 hr after a single bout of EE, at baseline and after 4 weeks. Our results demonstrate that markers of DNA damage or apoptosis were not affected by EE both in Control and ROJ group, and by ROJ, whereas, exercise temporarily affected the redox balance in both groups. Controls didn't change their response to EE after the experimental period, but experimental group after ROJ supplementation had lower EE-induced MDA, consumed less ascorbic acid, and had less activation of the hypoxanthine/xanthine system, i.e., they seemed to be protected from hypoxia/reoxygenation mechanisms.

Topics: Administration, Oral; Aged; Antioxidants; Ascorbic Acid; Beverages; Biomarkers; Citrus sinensis; Dietary Supplements; DNA Damage; Exercise; Female; Hemolysis; Homeostasis; Humans; Hypoxanthine; Malondialdehyde; Middle Aged; Oxidation-Reduction; Oxidative Stress; Physical Exertion; Plant Extracts; Xanthine

This study was performed to explore other potential mechanisms underlying hemolysis in addition to pore-formation of tentacle extract (TE) from the jellyfish Cyanea capillata. A dose-dependent increase of hemolysis was observed in rat erythrocyte suspensions and the hemolytic activity of TE was enhanced in the presence of Ca2+, which was attenuated by Ca2+ channel blockers (Diltiazem, Verapamil and Nifedipine). Direct intracellular Ca2+ increase was observed after TE treatment by confocal laser scanning microscopy, and the Ca2+ increase could be depressed by Diltiazem. The osmotic protectant polyethylenglycol (PEG) significantly blocked hemolysis with a molecular mass exceeding 4000 Da. These results support a pore-forming mechanism of TE in the erythrocyte membrane, which is consistent with previous studies by us and other groups. The concentration of malondialdehyde (MDA), an important marker of lipid peroxidation, increased dose-dependently in rat erythrocytes after TE treatment, while in vitro hemolysis of TE was inhibited by the antioxidants ascorbic acid-Vitamin C (Vc)-and reduced glutathione (GSH). Furthermore, in vivo hemolysis and electrolyte change after TE administration could be partly recovered by Vc. These results indicate that lipid peroxidation is another potential mechanism besides pore-formation underlying the hemolysis of TE, and both Ca2+ channel blockers and antioxidants could be useful candidates against the hemolytic activity of jellyfish venoms.

Topics: Animals; Antioxidants; Ascorbic Acid; Calcium; Calcium Channel Blockers; Cnidarian Venoms; Diltiazem; Erythrocyte Membrane; Erythrocytes; Glutathione; Hemolysis; Lipid Peroxidation; Male; Malondialdehyde; Nifedipine; Osmosis; Polyethylene Glycols; Rats; Rats, Sprague-Dawley; Scyphozoa; Verapamil

Topics: 2-Pyridinylmethylsulfinylbenzimidazoles; Ascorbic Acid; Cimetidine; Erythrocytes; Famotidine; Hemolysis; Histamine H2 Antagonists; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Lansoprazole; Lovastatin; Nizatidine; Omeprazole; Pantoprazole; Pravastatin; Proton Pump Inhibitors; Ranitidine; Ultraviolet Rays

Electron paramagnetic resonance (EPR) spectroscopy of spin labels was used to monitor membrane dynamic changes in erythrocytes subjected to oxidative stress with hydrogen peroxide (H(2)O(2)). The lipid spin label, 5-doxyl stearic acid, responded to dramatic reductions in membrane fluidity, which was correlated with increases in the protein content of the membrane. Membrane rigidity, associated with the binding of hemoglobin (Hb) to the erythrocyte membrane, was also indicated by a spin-labeled maleimide, 5-MSL, covalently bound to the sulfhydryl groups of membrane proteins. At 2% hematocrit, these alterations in membrane occurred at very low concentrations of H(2)O(2) (50 µM) after only 5 min of incubation at 37°C in azide phosphate buffer, pH 7.4. Lipid peroxidation, suggested by oxidative hemolysis and malondialdehyde formation, started at 300 µM H(2)O(2) (for incubation of 3 h), which is a concentration about six times higher than those detected with the probes. Ascorbic acid and α-tocopherol protected the membrane against lipoperoxidation, but did not prevent the binding of proteins to the erythrocyte membrane. Moreover, the antioxidant (+)-catechin, which also failed to prevent the cross-linking of cytoskeletal proteins with Hb, was very effective in protecting erythrocyte ghosts from lipid peroxidation induced by the Fenton reaction. This study also showed that EPR spectroscopy can be useful to assess the molecular dynamics of red blood cell membranes in both the lipid and protein domains and examine oxidation processes in a system that is so vulnerable to oxidation.

Topics: alpha-Tocopherol; Antioxidants; Ascorbic Acid; Catechin; Cyclic N-Oxides; Electron Spin Resonance Spectroscopy; Erythrocyte Membrane; Hemoglobins; Hemolysis; Humans; Hydrogen Peroxide; Hydrogen-Ion Concentration; Lipid Peroxidation; Membrane Fluidity; Membrane Proteins; Oxidative Stress; Thiobarbituric Acid Reactive Substances

Eryngium (E.) caucasicum was found as a new cultivated vegetable plant in northern Iran and used in several local foods. Little information is available on biological properties of E. caucasicum. In this work antioxidant activity of flavonoid-rich fraction of this plant was investigated by eight in vitro assay systems.. 1,1-diphenyl-2-picryl hydrazyl radical (DPPH), nitric oxide and hydrogen peroxide scavenging activities, Fe2+ chelating ability, reducing power and hemoglobin-induced linoleic acid peroxidation were used to evaluate antioxidant activities. Antihemolytic activities were evaluated against CuOOH and H2O2 induced hemolysis in rat erythrocyte.. IC50 for DPPH scavenging activity was 391.2 +/- 14.9, 706.6 +/- 22.3 and 779.7 +/- 16.7 microg ml(-1) for aqueous (AQ), ethyl acetate (EA) and n-hexane (HE) fractions, respectively. There was no difference between reducing power of AQ fraction activity and vitamin C (p > 0.05). IC50 for NO radical-scavenging activity was in order of AQ (133.5 +/- 6.2 microg ml(-1)) > EA (350.1 +/- 14.8 microg ml(-1)) > and HE (639.9 +/- 21.7 microg ml(-1)) fractions, respectively. Extracts showed weak Fe2+ chelating ability. HE fraction showed better activity (173.5 +/- 9.6 microg ml(-1)). Extracts exhibited weak hydrogen peroxide scavenging activity but exhibited very good antioxidant activity against the hemoglobin-induced linoleic acid peroxidation. There was no significant difference between AQ fraction and vitamin C (p > 0.01). Fractions delayed the onset of the CuOOH induced hemolysis. AQ fraction showed very high total phenol and flavonoid contents which was higher than other fractions. High phenol and flavonoid content of AQ fraction may lead to its better antioxidant activity.. E. caucasicum fractions exhibited different levels of antioxidant and antihemolytic activities in all tested models. These results can be useful as a starting point of view for further applications of E. caucasicum aerial parts or its constituents in pharmaceutical preparations.

Topics: Animals; Antioxidants; Ascorbic Acid; Dose-Response Relationship, Drug; Eryngium; Erythrocytes; Flavonoids; Hemolysis; Hydrogen Peroxide; Inhibitory Concentration 50; Male; Phenols; Plant Extracts; Rats; Rats, Wistar; Solvents

Artemisia (Asteraceae) contains more than 400 species. Many of the plants belonging to this genus are known to possess biological properties. In this study, antioxidant and antihemolytic activities of flavonoid rich fractions of A. tschernieviana Besser were evaluated.. Plant aerial parts were extracted with 60% acetone. Extract was fractionated sequentially with hexane (HE), ethyl acetate (EA) and water (AQ). Antioxidant and antihemolytic activities of these fractions were assessed. Their antihemolytic activity was determined by H2O2 and cumene hydroperoxide induced hemolysis models.. AQ fraction showed very powerful activity in 1,1-disphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity with IC50 = 0.12 +/- 0.01 microg ml(-1) that was better than controls (vitamin C, quercetin and butylated hydroxyanisole (BHA)). This fraction showed very powerful reducing power assay that was better than vitamin C (p < 0.01). EA fraction showed higher activity in scavenging nitric oxide with IC50 = 0.14 +/- 0.04 microg ml(-1). In iron chelating activity HE fraction showed the best activity (IC50 = 162.2 +/- 8.7 microg ml(-1)). In scavenging of hydrogen peroxide, AQ fraction showed better activity than control group. This fraction had higher phenol and flavonoid contents. EA fraction showed higher antihemolytic activity with IC50 = 728.8 +/- 29 microg ml(-1).. The fractions show very good activities in studied models. Aqueous fractions showed better activity than the others in nearly all tested models. These results can be useful as a starting point of view for further applications of A. tschernieviana aerial parts or its constituents in pharmaceutical preparations after performing clinical in vivo researches.

Topics: Animals; Antioxidants; Artemisia; Ascorbic Acid; Butylated Hydroxyanisole; Erythrocytes; Flavonoids; Hemolysis; Inhibitory Concentration 50; Male; Nitric Oxide; Phenols; Plant Components, Aerial; Plant Extracts; Quercetin; Rats; Rats, Wistar; Solvents

Naphthalene, a widely used industrial and household chemical, has rarely been an agent of poisoning worldwide. Severe haemolysis from naphthalene poisoning is rare and can be a challenge to clinicians. We report a 22-year-old female, who accidentally ingested naphthalene mixed coconut oil and got admitted with recurrent vomiting, headache and passage of dark urine. Severe intravascular haemolysis with hypotension and neutrophilic leukocytosis was detected. She was treated with red blood cell transfusions, intravenous saline infusion and ascorbic acid.

Topics: Administration, Oral; Anemia, Hemolytic; Ascorbic Acid; Coconut Oil; Erythrocyte Transfusion; Female; Glucose; Hemoglobinuria; Hemolysis; Humans; Hypotension; Infusions, Intravenous; Methemoglobinemia; Naphthalenes; Plant Oils; Poisoning; Severity of Illness Index; Treatment Outcome; Young Adult

Experiments were performed with the aim of investigating the effect of packing on erythrocyte osmotic fragility (EOF) and malondialdehyde (MDA) concentration in donkeys, and the effect of ascorbic acid (AA). Twelve apparently healthy donkeys raised under the traditional extensive system served as experimental subjects. Six donkeys administered orally with AA (200 mg/kg) and subjected to packing were used as experimental animals, whilst six others not administered with AA served as controls. Blood samples were collected pre- and post-packing from all the donkeys for the determination of MDA and EOF. At 0.3% Sodium Chloride (NaCl) concentration, the percentage haemolysis was 93.69% ± 2.21% in the control donkeys and the value was significantly (P < 0.05) higher than the value of 71.31% ± 8.33%, recorded in the experimental donkeys. The post-packing MDA concentration obtained in the control donkeys was 39.62 µmol ± 4.16 µmol, and was not significantly different (P > 0.05) from the value of 35.97 µmol ± 2.88 µmol recorded in the experimental donkeys. In conclusion, the increase in haemolysis obtained in the donkeys suggested that packing induced oxidative stress, which was ameliorated by AA administration.

Topics: Animals; Ascorbic Acid; Case-Control Studies; Equidae; Erythrocytes; Female; Hemolysis; Male; Malondialdehyde; Osmotic Fragility; Oxidative Stress; Sodium Chloride

Three new diterpenes Amijiol acetate (3), dolabellane, Dolabellatrienol (4), and dolastane, Amijiol-7, 10-diacetate (9) were isolated together with the previously described Pachydictyol A (1), Isopachydictyol A (2), 8β-hydroxypachydictyol A (5), Amijiol (6), Isodictyohemiacetal (7) and Dictyol C (8) from the Red Sea brown alga Dictyota dichotoma var. implexa. The structures and relative stereochemistry of the new diterpenoids were proposed on the basis of their spectral data. Compounds 3 and 9 have potent activity against DNA damage, cytotoxicity against WI-38, HepG2, and MCF-7 cell lines, and antioxidant using ABTS and erythrocytes hemolysis.

Topics: Animals; Antineoplastic Agents; Antioxidants; Benzothiazoles; Cell Line, Tumor; Cell Survival; Chlorocebus aethiops; Diterpenes; DNA Damage; Erythrocytes; Hemolysis; Humans; Phaeophyceae; Sulfonic Acids; Vero Cells

In the present work, 1-(5-methyl-2-phenyl-1H-imidazol-4-yl)ethanone 1 was prepared and used as a precursor for the synthesis of new thiazole, arylidiene and coumarin derivatives. The antimicrobial, antioxidant, anti-hemolytic, and cytotoxic activities of new compounds have been screened. Compound 12 showed an excellent antibacterial activity for all the tested bacteria with minimal inhibitory concentration (MIC) ranged between 21.9 and 43.8 μg/mL. While, compounds 2, 8 and 10a were the best antioxidant reagents using the DPPH method. Compounds 6a and 10b proved to exhibit potent antioxidative activity as reflected in the ability to inhibit lipid per-oxidation in rat brain and kidney homogenates and rate erythrocyte hemolysis. Compounds 6a proved to have the highest cytotoxic activity (81.9%) followed by 2, 6c, 7b and 12 using in vitro Ehrlich ascites assay. The details synthetic methods, spectroscopic data and biological results are recorded.

Topics: Animals; Anti-Bacterial Agents; Antineoplastic Agents; Antioxidants; Bacteria; Cell Proliferation; Drug Screening Assays, Antitumor; Erythrocytes; Hemolysis; Heterocyclic Compounds; Imidazoles; Microbial Sensitivity Tests; Molecular Structure; Rats; Stereoisomerism; Structure-Activity Relationship

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

Phytochemicals such as polyphenols and carotenoids are gaining importance because of their contribution to human health and their multiple biological effects such as antioxidant, antimutagenic, anticarcinogenic, and cytoprotective activities and their therapeutic properties. Banana peel is a major by-product in pulp industry and it contains various bioactive compounds like polyphenols, carotenoids, and others. In the present study, effect of ripening, solvent polarity on the content of bioactive compounds of crude banana peel and the protective effect of peel extracts of unripe, ripe, and leaky ripe banana fruit on hydrogen peroxide-induced hemolysis and their antioxidant capacity were investigated. Banana (Musa paradisica) peel at different stages of ripening (unripe, ripe, leaky ripe) were treated with 70% acetone, which were partitioned in order of polarity with water, ethyl acetate, chloroform (CHCl₃), and hexane sequentially. The antioxidant activity of the samples was evaluated by the red cell hemolysis assay, free radical scavenging (1,1-diphenyl-2-picrylhydrazyl free radical elimination) and superoxide dismutase activities. The Folin-Ciocalteu's reagent assay was used to estimate the phenolic content of extracts. The findings of this investigation suggest that the unripe banana peel sample had higher antioxidant potency than ripe and leaky ripe. Further on fractionation, ethyl acetate and water soluble fractions of unripe peel displayed high antioxidant activity than CHCl₃ and hexane fraction, respectively. A positive correlation between free radical scavenging capacity and the content of phenolic compound were found in unripe, ripe, and leaky ripe stages of banana peel.

Topics: Antioxidants; Ascorbic Acid; Biotechnology; Biphenyl Compounds; Erythrocytes; Flavonoids; Food Handling; Free Radical Scavengers; Free Radicals; Fruit; Hemolysis; Humans; Musa; Oxidation-Reduction; Phenols; Picrates; Plant Extracts; Polyphenols; Solvents; Superoxide Dismutase

6-O-alkyl ascorbic acid esters (ASC(n)) are amphiphilic molecules that behave as surfactants in aqueous solution. ASC(n) have shown some physical and rheological properties that suggest a potential utility as drug carriers. The present paper aims to evaluate the effect of ASC(n) on erythrocyte properties in order to get information regarding the relationship between osmotic fragility, erythrocyte deformability and membrane lipoperoxidation process. The assays were performed at the following concentrations: the critical micelar concentration (CMC), producing 10% hemolysis (CH(10)) and producing 50% hemolysis (CH(50)). We observed that ASC(n) (ASC(8), ASC(10) and ASC(12)), at concentration nearby CMC, neither affected cell deformability nor produced lipoperoxidation. Nevertheless, at higher concentrations (CH(10) and CH(50)), the RBCs incubated with ASC(n) were affected by a significant and progressive loss of deformability, simultaneously with an increase of osmotic fragility and membrane lipoperoxidation.

Topics: Adult; Ascorbic Acid; Dose-Response Relationship, Drug; Erythrocyte Deformability; Erythrocyte Membrane; Esters; Fatty Acids; Hemolysis; Humans; Lipid Peroxidation; Osmotic Fragility; Oxidative Stress

2-O-β-D-Glucopyranosyl-L-ascorbic acid (AA-2βG) is a natural derivative of vitamin C (Lascorbic acid, AA) isolated from Goji berry (Lycium barbarum L.) fruit. We evaluated the antioxidant activities of AA-2βG and AA using in vitro and in vivo model systems. In vitro radical scavenging assays demonstrated that AA-βG was capable of scavenging 1,1-diphenyl-2-picryl-hydrazyl and hydroxyl peroxide and inhibiting H(2)O(2)-induced hemolysis better than AA. AA-2βG and AA had similar hydroxyl radical scavenging capabilities, but AA-2βG was incapable of scavenging superoxide anion radicals, and its capacity to scavenge nitrite (NO(2) (-)) was lower than that of AA. The overall in vitro reduction capability of AA-2βG was also significantly lower than that of AA. Moreover, in vivo studies demonstrated that AA-2βG was capable of protecting the liver against carbon tetrachloride-induced acute liver injury in mice. These results suggest that AA-2βG is an important antioxidant component of Goji berry fruit, which may share similar but distinct antioxidant mechanistic properties with AA. This study furthers our understanding of the mechanisms of Goji berry fruit pharmacological activities on antiaging and antitumor properties as a traditional medicine and dietary supplement.

Topics: Animals; Antioxidants; Ascorbic Acid; Carbon Tetrachloride; Chemical and Drug Induced Liver Injury; Dietary Supplements; Free Radical Scavengers; Fruit; Hemolysis; Hydrogen Peroxide; Kinetics; Lycium; Male; Mice; Mice, Inbred Strains; Oxidants; Oxidation-Reduction; Oxidative Stress; Rabbits; Random Allocation

Ferula gummosa Boiss is native to central Asia. This plant has traditionally been used in the treatment of many diseases. The antihypoxic and antioxidant activities of Ferula gummosa roots were investigated.. 1,1-diphenyl-2-picryl hydrazyl radical (DPPH), nitric oxide and hydrogen peroxide scavenging activities, Fe2+ chelating ability, reducing power and hemoglobin-induced linoleic acid peroxidation were used to evaluate antioxidant activities. Antihemolytic activity was evaluated by H2O2 induced hemolysis in rat erythrocytes. The total amount of phenolic compounds was determined as gallic acid equivalents and total flavonoid contents were calculated as quercetin equivalents from a calibration curve.. The extracts showed moderate antioxidant activity in some models. IC50 for DPPH radical-scavenging activity was 579.6 +/- 19.4 microg/ml. The extracts showed weak nitric oxide-scavenging activity between 0.1 and 1.6 mg ml(-1) but showed good Fe2+ chelating ability. IC50 was 895.5 +/- 24.1 microg/ml. The extract also exhibited low antioxidant activity in the linoleic acid model but were capable of scavenging hydrogen peroxide in a concentration dependent manner. Tested extract show moderate activity in H2O2 induced hemolysis in rat erythrocytes which was not comparable with vitamin C.. F. gummosa Boiss root showed different level antioxidant and antihemolytic activities. Biological effects may be attributed, at least in part, to the presence of phenols and flavonoids in the extract.

Topics: Animals; Antioxidants; Ascorbic Acid; Dose-Response Relationship, Drug; Erythrocytes; Ferula; Flavonoids; Free Radical Scavengers; Hemolysis; Hydrogen Peroxide; Inhibitory Concentration 50; Male; Medicine, East Asian Traditional; Nitric Oxide; Phenols; Plant Extracts; Plant Roots; Rats; Rats, Wistar

In the present paper the photodynamic effect of hypericin on superoxide dismutase activity and the possibility of reduction of hypericin phototoxicity by antioxidants were studied. It was shown an almost twice decrease in superoxide dismutase activity of red blood cells under the photosensitization by hypericin. The influence of antioxidants (ascorbic acid and quercetin) on hypericin photodynamic action has revealed that these antioxidants suppress or stimulate photohemolysis caused by hypericin. The photosensitization reaction realized by hypericin could be shifted from type II to type I or vice versa by manipulating the antioxidant concentration. Strengthening of photohemolysis by antioxidants in some concentrations indicates the switching of alternative mechanisms of hypericin photodynamic action and its complicated manner. Thus the selection of antioxidant concentrations is of extreme importance for changing the efficacy of photodynamic therapy with hypericin.

Topics: Algorithms; Anthracenes; Antioxidants; Ascorbic Acid; Calcium Channels; Dose-Response Relationship, Drug; Erythrocytes; Hemolysis; Humans; Male; Peroxidases; Perylene; Photosensitizing Agents; Protein Kinase C; Quercetin; Reactive Oxygen Species; Superoxide Dismutase; Time Factors

The aim of our study was to compare and assess the effectiveness of antioxidant mixtures on the erythrocytes (RBC) of adult male albino rats (Wister) subjected to simulated intermittent high altitudes--5,100 m (AL(1)) and 6,700 m (AL(2))--to induce oxidative stress (OS). To achieve our objective, we pre-supplemented four sets of animals with different antioxidant mixtures [vitamin E (vit.E; 50 IU/kg BW), vitamin C (vit.C; 400 mg/kg) and L: -carnitine (400 mg/kg)] in different combinations [M1 (vit.E+vit.C), M2 (vit.C+carnitine), M3 (vit.E+carnitine) and M4 (vit.C+vit.E+carnitine)] for 30 days prior to as well during exposure to intermittent hypobaric hypoxia (IHH). Membrane instability, in terms of osmotic fragility and hemolysis, decreased in RBCs of supplemented animals. There was a significant increase in the activity of glutathione peroxidase in the RBCs of supplemented animals. We confirmed OS imposed by IHH with assays relating to lipid [thiobarbituric acid reactive substances (TBARS) and lipofuscin (LF)] and protein (carbonyl, PrC) oxidation, and found a positive correlation between PrC and hemolysis, with a decrease in both upon supplementation with M3 and M4 mixtures. Fluorescence microscopic observation showed a maximum decrease in the LF content in rats administered M4 and M1 compared to those on M2 and M3 mixtures at both altitudes. We suggest that multiple antioxidant fortifications are effective in overcoming increased OS experienced by RBCs at high altitudes.

Topics: Altitude; Animals; Antioxidants; Ascorbic Acid; Carnitine; Erythrocytes; Glutathione Peroxidase; Hemolysis; Hypoxia; Lipid Peroxidation; Male; Microscopy, Fluorescence; Oxidative Stress; Rats; Rats, Wistar; Time Factors; Vitamin E

The halo 6-fatty acid esters of L-ascorbic acid 3a, 3b and 6-fatty acid esters of L-ascorbic acid 5a-g were achieved from L-ascorbic acid 1. Compounds 3a, 3b and 5a-g were evaluated for anti-oxidant, anti-lipid peroxidation, and secretory phospholipase A(2) (sPLA(2)) inhibition in vitro, and sPLA(2) induced mouse paw edema. All the derivatives retained their anti-oxidant property compared to ascorbic acid at 6 × 10(-4)M and are good inhibitors of lipid peroxidation at 1 mg ml(-1) as evaluated by 2, 2-Diphenyl-1-picrylhydrazyl radical and thio-barbituric acid methods, respectively. Compounds 5e and 5f significantly inhibited purified group I sPLA(2) from Naja naja and group II sPLA(2) from Vipera russelli, human synovial fluid and human pleural fluid with IC(50) value ranging from 64 ± 1.95 to 82 ± 1.3 and 48 ± 2.27 to 61 ± 2.23 μM, respectively. The compounds 5e and 5f also showed varying degree of potency in neutralizing indirect hemolytic activity of sPLA(2) at 50 μM concentration, and sPLA(2) induced mouse paw edema at the dose 3 mg/kg. Further docking studies also confirmed that compounds 5e and 5f have maximum interaction with increasing negative energy value. Single molecule possessing both anti-oxidant and anti-inflammatory activities is of great therapeutic significance in inflammatory disorders.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Ascorbic Acid; Drug Evaluation, Preclinical; Edema; Fatty Acids; Hemolysis; Humans; Hydrophobic and Hydrophilic Interactions; Lipid Peroxidation; Mice; Phospholipases A2, Secretory; Protein Binding; Snake Venoms; Structure-Activity Relationship

The 5,10-dihydro-2-thioxo-pyrimido[4,5-b]quinolines (2a-c) and its oxidized form 3 were prepared and used as key intermediates for the synthesis of thiazolo[3',2':1,2]pyrimido[4,5-b]-quinolines (5a-c), isoxazolo[5'',4'':4',5']thiazolo[3',2':1,2]pyrimido[4,5-b]quinolines (6a-c), 4-chloro-2-methylthio-pyrimido[4,5-b]quinoline, its amino derivatives (19-21) and 10,11,12,13-tetrahydro-5H-quino[2',3':4,5]pyrimido[6,1-b]quinazoline (22). The newly synthesized compounds were characterized by IR, NMR (1H, 13C) and mass spectral studies. Representative of the synthesized compounds was tested and evaluated for anti-oxidant, anti-inflammatory and analgesic activities. Compounds 2a-c showed the highest inhibitory anti-oxidant activity either using erythrocyte hemolysis or ABTS methods. Compounds 2a, 10b, 16, and 17a manifested the best protective effect against DNA damage induced by bleomycin. Compounds 2c, 5a, 20a, 2a, and 2b exhibited a potent anti-inflammatory activity using carrageenan-induced paw edema test in rats.

Topics: Analgesics; Animals; Anti-Inflammatory Agents; Antioxidants; Brain; DNA Damage; Drug Design; Edema; Erythrocytes; Hemolysis; Kidney; Quinolines; Rats; Spectrum Analysis

Ingestional naphthalene mothball poisoning leading to prolonged haemolysis and methaemoglobinaemia can present with diagnostic and therapeutic challenges. A 19-year-old woman ingested 12 mothballs, and presented two days later with haemolysis and methaemoglobinaemia. She was treated with red blood cell transfusions, intravenous methylene blue, N-acetylcysteine and ascorbic acid. Continuous venovenous haemofiltration was conducted for 45 hours. Haemolysis with anaemia and methaemoglobinaemia persisted even after five days post-ingestion. Clinical and biochemical parameters improved. We describe a case of ingestional naphthalene poisoning with a good outcome after treatment.

Topics: Acetylcysteine; Adult; Anemia, Hemolytic; Ascorbic Acid; Erythrocyte Transfusion; Female; Hemolysis; Humans; Methemoglobinemia; Methylene Blue; Naphthalenes; Poisoning; Suicide, Attempted; Time Factors; Treatment Outcome

The antioxidant activity of a provitamin C agent, 2-O-beta-D-glucopyranosyl-L-ascorbic acid (AA-2betaG), was compared to that of 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G) and ascorbic acid (AA) using four in vitro methods, 1,1-diphenyl-picrylhydrazyl (DPPH) radical-scavenging assay, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS(*+))-scavenging assay, oxygen radical absorbance capacity (ORAC) assay, and 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced erythrocyte hemolysis inhibition assay. AA-2betaG slowly and continuously scavenged DPPH radicals and ABTS(*+) in roughly the same reaction profiles as AA-2G, whereas AA quenched these radicals immediately. In the ORAC assay and the hemolysis inhibition assay, AA-2betaG showed similar overall activities to AA-2G and to AA, although the reactivity of AA-2betaG against the peroxyl radical generated in both assays was lower than that of AA-2G and AA. These data indicate that AA-2betaG had roughly the same radical-scavenging properties as AA-2G, and a comprehensive in vitro antioxidant activity of AA-2betaG appeared to be comparable not only to that of AA-2G but also to that of AA.

Topics: Amidines; Animals; Antioxidants; Ascorbic Acid; Benzothiazoles; Erythrocytes; Hemolysis; Hydrogen-Ion Concentration; Molecular Structure; Sheep; Sulfonic Acids

Oxidative damage of biomolecules and antioxidant status in erythrocytes of humans from an outbreak of argemone oil (AO) poisoning in Kannauj (India) and AO intoxicated experimental animals was investigated. Erythrocytes of the dropsy patients and AO treated rats were found to be more susceptible to 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) induced peroxidative stress. Significant decrease in RBC glutathione (GSH) levels (46, 63%) with concomitant enhancement in oxidized glutathione (172, 154%) levels was noticed in patients and AO intoxicated animals. Further, depletion of glutathione reductase (GR), glucose-6-phosphate dehydrogenase (G-6-PDH) and glutathione-S-transferase (GST) (42-52%) was observed in dropsy patients. Oxidation of erythrocyte membrane lipids and proteins was increased (120-144%) in patients and AO treated animals (112-137%) along with 8-OHdG levels in whole blood (180%) of dropsy patients. A significant reduction in alpha-tocopherol content (68%) was noticed in erythrocytes of dropsy patients and hepatic, plasma and RBCs of AO treated rats (59-70%) thereby indicating the diminished antioxidant potential to scavenge free radicals or the limited transport of alpha-tocopherol from liver to RBCs leading to enhanced oxidation of lipids and proteins in erythrocytes. These studies implicate an important role of erythrocyte degradation in production of anemia and breathlessness in epidemic dropsy.

Topics: Adolescent; Adult; Aged; Animals; Antioxidants; Ascorbic Acid; Child; Child, Preschool; Erythrocytes; Female; Glutathione; Hemolysis; Humans; Male; Middle Aged; Plant Oils; Rats; Rats, Wistar; Vitamin E

New 7-aryl or 7-heteroarylamino-2,3-dimethylbenzo[b]thiophenes were prepared by palladium-catalyzed Buchwald-Hartwig cross-coupling of 7-bromo or 7-amino-2,3-dimethylbenzo[b]thiophenes, previously prepared by us, with substituted (4-methoxy or 3,4-dimethoxy) anilines and 3-aminopyridine or with substituted (3-methoxy or 4-cyano) bromobenzenes and 2-bromopyridine, respectively, using Pd(OAc)2, rac-BINAP or Xantphos as ligands, and Cs2CO3 as base. Their antioxidant properties were evaluated by several methods: reducing power, scavenging effect on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals, inhibition of erythrocyte hemolysis and inhibition of lipid peroxidation using the beta-carotene linoleate system. EC50 values for all the methods were determined and it was possible to establish some structure-activity relationships (SARs) based on the presence and position of different substituents on the phenyl ring (1 or 2 OMe and CN), on the presence of a pyridine ring and on the position of its nitrogen atom relative to the N-H bond.

Topics: Antioxidants; Cross-Linking Reagents; Free Radical Scavengers; Hemolysis; Humans; Inhibitory Concentration 50; Lipid Peroxidation; Structure-Activity Relationship; Thiophenes

6-O-alkyl ascorbic acid esters (ASCn) are amphiphilic molecules that behave as surfactants in aqueous solution. These compounds show physico-chemical and aggregation properties that depend on the alkyl chain length, pH and temperature. It must consider that ASCn have shown some physical and rheological properties that suggest a potential utility as drug carriers. The present paper aims to evaluate the effects of these surfactants on human erythrocyte membranes. The membrane properties studied were: osmotic resistance in hypotonic media, shape transformation, and vesicle release at lytic concentration. According to our results, all properties depended on the length of the hydrophobic chain and they did not evolve monotonically. Finally, the study of ASCn interaction with erythrocyte membrane allowed us to postulate the crucial influence that the molecular structure exerts upon the manner in which amphiphiles interact with biological membranes and the effects involved in them.

Topics: Ascorbic Acid; Erythrocyte Deformability; Erythrocyte Membrane; Esters; Hemolysis; Humans; Hypotonic Solutions; Osmotic Fragility; Structure-Activity Relationship; Surface-Active Agents

Phenyldrazine (PHDZ) provokes acute haemolysis and generates free radicals that are known to be involved in the toxicity of the drug. Vitamin C and E have been ascribed beneficial roles in many acute haemolytic states and oxidative stress.. The present study was designed to assess the antioxidant effect of vitamin C and E on PHDZ- induced haemolysis in SD rat in-vivo and ex-vivo.. Rats treated for weeks with oral vitamin E (500mg/kg/day, n=6), oral vitamin C (100mg/kg/day, n=6) and control rats, which received tap water; were monitored for peripheral blood parameters and haemolytic activity of erythrocytes at Baseline, 48 hours and 1 week respectively after intraperitonial administration of PHDZ (15mg/kg/day for 7 days). Osmotic fragility as well as organ weights were determined 1 week after PHDZ. Dose-dependent effects of vitamins C and E on haemolysis were determined at the end of 120 min incubation of rat RBC with the vitamins in phosphate-buffered-saline.. Hb and PCV were better preserved in rats pre-treated with the antioxidants. The spleenomegaly, cardiomegaly and hepatomegaly associated with PHDZ treatment were significantly reduced in antioxidant pre-treated rats (P<0.001). reduction in cardiac weight was more marked in vitamin E group (0.56+/- 0.02g vs. 0.72+/-0.03g; n=6; P<0.01; vitamin E vs. PHDZ) compared to vitamin C (0.69+/-0.02g vs. 0.72+/-0.03g; n=6; P>0.05; vitamin C compared to PHDZ). RBC of vitamin E group exhibited better resistance to osmoticlysis (P<0.001). at higher doses (400-500mg/L), vitamin C caused significant haemolysis both in the absence (P<0.001) and presence of PHDZ (P<0.001). Vitamin E abolished the long tail of the osmotic fragiligram.. The result of this study suggests that compared to vitamin C, vitamin E may better protect RBC against haemolysis induced by oxidative stress.

Topics: Animals; Antioxidants; Ascorbic Acid; Erythrocytes; Hemolysis; Humans; Nigeria; Oxidative Stress; Phenylhydrazines; Rats; Rats, Sprague-Dawley; Vitamin E; Vitamins

Vitamin C is a popular antioxidant; however, its water solubility limits its function in the lipid environment. As a result, the antioxidative properties of its lipophilic derivatives have aroused research attention, especially L-ascorbyl-6-laurate (VC-12). We have investigated the effect of a high concentration of VC-12 on 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH)-induced haemolysis of human erythrocytes. The findings indicated that VC-12 was capable of protecting erythrocytes against AAPH-induced haemolysis when its concentration was below 60 microM. With an increase in the concentration of VC-12 and a decrease in the concentration of AAPH, VC-12 promoted haemolysis remarkably, the mechanism of which has been proposed as VC-12-mediated peroxidation. When the concentration of VC-12 was increased to above 150 microM, VC-12 promoted haemolysis by its function as a surfactant, to unbalance the osmotic pressure within and outside erythrocytes. High concentrations of VC-12 may have generated radicals via autooxidation, resulting in eventual haemolysis. Therefore, the double-faced effect of VC-12 on haemolysis of human erythrocytes was due to its concentration. This information may be useful for the safe use of lipophilic vitamin C.

Topics: Amidines; Antioxidants; Ascorbic Acid; Dose-Response Relationship, Drug; Erythrocytes; Free Radicals; Hemolysis; Humans; Lipid Peroxidation; Osmotic Pressure; Oxidants; Oxidation-Reduction

Inhibitory effects of 2-O-substituted ascorbic acid derivatives, ascorbic acid 2-glucoside (AA-2G), ascorbic acid 2-phosphate (AA-2P), and ascorbic acid 2-sulfate (AA-2S), on 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of sheep erythrocytes were studied and were compared with those of ascorbic acid (AA) and other antioxidants. The order of the inhibition efficiency was AA-2S> or =Trolox=uric acid> or =AA-2P> or =AA-2G=AA>glutathione. Although the reactivity of the AA derivatives against AAPH-derived peroxyl radical (ROO(*)) was much lower than that of AA, the derivatives exerted equal or more potent protective effects on AAPH-induced hemolysis and membrane protein oxidation. In addition, the AA derivatives were found to react per se with ROO(*), not via AA as an intermediate. These findings suggest that secondary reactions between the AA derivative radical and ROO(*) play a part in hemolysis inhibition. Delayed addition of the AA derivatives after AAPH-induced oxidation of erythrocytes had already proceeded showed weaker inhibition of hemolysis compared to that of AA. These results suggest that the AA derivatives per se act as biologically effective antioxidants under moderate oxidative stress and that AA-2G and AA-2P may be able to act under severe oxidative stress after enzymatic conversion to AA in vivo.

Topics: Amidines; Animals; Antioxidants; Ascorbic Acid; Erythrocyte Membrane; Free Radicals; Hemolysis; Sheep; Time Factors

In the present work, we investigated the effect of ascorbic acid and glutathione on hemolysis induced by hemin in erythrocytes. Ascorbic acid not only enhanced hemolysis, but also induced formation of thiobarbituric acid-reactive substances in the presence of hemin. It has been shown that glutathione inhibits hemin-induced hemolysis by mediating hemin degradation. Erythrocytes depleted of glutathione became very sensitive to oxidative stress induced by hemin and ascorbic acid. H(2)O(2) was involved in hemin-mediated hemolysis in the presence of ascorbic acid. However, a combination of glutathione and ascorbic acid was more effective in inhibiting hemolysis induced by hemin than glutathione alone. Extracellular and intracellular ascorbic acid exhibited a similar effect on hemin-induced hemolysis or inhibition of hemin-induced hemolysis by glutathione. The current study indicates that ascorbic acid might function as an antioxidant or prooxidant in hemin-mediated hemolysis, depending on whether glutathione is available.

Topics: Ascorbic Acid; Cells, Cultured; Dose-Response Relationship, Drug; Drug Combinations; Erythrocytes; Glutathione; Hemin; Hemolysis; Humans

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

Following oxidative stress, modifications of several biologically important macromolecules have been demonstrated. In this study we investigated the effect of a natural extract from Mangifera indica L (Vimang), its main ingredient mangiferin and epigallocatechin gallate (EGCG) on energy metabolism, energy state and malondialdehyde (MDA) production in a red blood cell system. Analysis of MDA, high energy phosphates and ascorbate was carried out by high performance liquid chromatography (HPLC). Under the experimental conditions, concentrations of MDA and ATP catabolites were affected in a dose-dependent way by H2O2. Incubation with Vimang (0.1, 1, 10, 50 and 100 microg/mL), mangiferin (1, 10, 100 microg/mL) and EGCG (0.01, 0.1, 1, 10 microM) significantly enhances erythrocyte resistance to H2O2-induced reactive oxygen species production. In particular, we demonstrate the protective activity of these compounds on ATP, GTP and total nucleotides (NT) depletion after H2O2-induced damage and a reduction of NAD and ADP, which both increase because of the energy consumption following H2O2 addition. Energy charge potential, decreased in H2O2-treated erythrocytes, was also restored in a dose-dependent way by these substances. Their protective effects might be related to the strong free radical scavenging ability described for polyphenols.

Topics: Ascorbic Acid; Catechin; Chromatography, High Pressure Liquid; Erythrocytes; Hemolysis; Humans; Hydrogen Peroxide; Lipid Peroxidation; Malondialdehyde; Mangifera; Plant Extracts; Xanthones

Assessment of the antioxidant activity of vitamins and other compounds is of interest in the understanding of their in vivo effects. In this study, we have investigated the activity of several lipid and water-soluble vitamins in human whole blood. Measurements were carried out using a biological test that enables the evaluation of both red blood cells and plasma resistance against free radical activity induced by 2,2'-azobis (2-amidinopropane)hydrochloride (AAPH). Antioxidant activity of vitamins has been determined by using the biological test versus chemical methods (chemiluminescence, DMPD radical). We have observed strong anti-oxidant potentials for vitamins B6 and B9 with biological tests, but not with chemical methods. At 10 microM, the vitamin B9 efficiency in inhibiting radical-induced red blood cell hemolysis was almost three times higher than vitamin C efficiency and two times higher than alpha-tocopherol efficiency. Antioxidant activity was not observed for vitamins B1 or B2, nor for retinol. The weak activity of beta-carotene still remains to be investigated particularly in relation to oxygen pressure. Our study demonstrated that the biological test is more useful than the chemical methods employed in this instance, for the evaluation of antioxidant capacity of lipophilic and putatively biologically active compounds.

Topics: alpha-Tocopherol; Amidines; Antioxidants; Ascorbic Acid; beta Carotene; Biological Assay; Caffeic Acids; Dose-Response Relationship, Drug; Fluorescent Dyes; Hemolysis; Humans; Luminescent Measurements; Oxidants; Phenylenediamines; Reference Values; Solubility; Vitamin A; Vitamin B Complex; Vitamins

Oxidative stress seems to contribute to cardiopulmonary bypass (CPB)-related postoperative complications. Pediatric patients are particularly prone to these complications. With this in mind, we measured oxidative stress markers in blood plasma of 20 children undergoing elective heart surgery before, during, and up to 48 h after cessation of CPB, along with inflammatory parameters and full analysis of iron status. Ascorbate levels were decreased by approximately 50% (P < 0.001) at the time of aorta cross-clamp removal (or pump switch-off in 4 patients with partial CPB), and associated with corresponding increases in dehydroascorbate (P < 0.001, r = -0.80) and malondialdehyde (P < 0.01, r = -0.59). In contrast to the immediate oxidative response, peak levels of IL-6 and IL-8 were not observed until 3-12 h after CPB cessation. The early loss of ascorbate correlated with duration of CPB (P < 0.002, r = 0.72), plasma hemoglobin after cross-clamp removal (P < 0.001, r = 0.70), and IL-6 and IL-8 levels at 24 and 48 h after CPB (P < 0.01), but not with postoperative lactate levels, strongly suggesting that hemolysis, and not inflammation or ischemia, was the main cause of early oxidative stress. The correlation of ventilation time with early changes in ascorbate (P < 0.02, r = 0.55), plasma hemoglobin (P < 0.01, r = 0.60), and malondialdehyde (P < 0.02, r = 0.54) suggests that hemolysis-induced oxidative stress may be an underlying cause of CPB-associated pulmonary dysfunction. Optimization of surgical procedures or therapeutic intervention that minimize hemolysis (e.g., off-pump surgery) or the resultant oxidative stress (e.g., antioxidant treatment) should be considered as possible strategies to lower the rate of postoperative complications in pediatric CPB.

Topics: Ascorbic Acid; C-Reactive Protein; Cardiac Surgical Procedures; Cardiopulmonary Bypass; Child; Child, Preschool; Dehydroascorbic Acid; Heart Defects, Congenital; Hemolysis; Humans; Infant; Interleukin-6; Interleukin-8; Iron; Ischemia; Malondialdehyde; Neutrophils; Oxidative Stress; Pneumonia; Postoperative Complications; Prospective Studies

Intravascular hemolysis is one of the most emphasized mechanisms for destruction of erythrocytes during and after physical activity. Exercise-induced oxidative stress has been proposed among the different factors for explaining exercise-induced hemolysis. The validity of oxidative stress following exhaustive cycling exercise on erythrocyte damage was investigated in sedentary and trained subjects before and after antioxidant vitamin treatment (A, C, and E) for 2 mo. Exercise induced a significant increase in thiobarbituric acid-reactive substance and protein carbonyl content levels in sedentary subjects and resulted in an increase of osmotic fragility and decrease in deformability of erythrocytes, accompanied by signs for intravascular hemolysis (increase in plasma hemoglobin concentration and decrease in haptoglobulin levels). Administration of antioxidant vitamins for 2 mo prevented exercise-induced oxidative stress (thiobarbituric acid-reactive substance, protein carbonyl content) and deleterious effects of exhaustive exercise on erythrocytes in sedentary subjects. Trained subjects' erythrocyte responses to exercise were different from those of sedentary subjects before antioxidant vitamin treatment. Osmotic fragility and deformability of erythrocytes, plasma hemoglobin concentration, and haptoglobulin levels were not changed after exercise, although the increased oxidative stress was observed in trained subjects. After antioxidant vitamin treatment, functional and structural parameters of erythrocytes were not altered in the trained group, but exercise-induced oxidative stress was prevented. Increased percentage of young erythrocyte populations was determined in trained subjects by density separation of erythrocytes. These findings suggest that the exercise-induced oxidative stress may contribute to exercise-induced hemolysis in sedentary humans.

Topics: Adult; Antioxidants; Ascorbic Acid; Bicycling; Erythrocyte Deformability; Erythrocytes; Exercise; Hemolysis; Humans; Life Style; Male; Osmotic Fragility; Oxidative Stress; Oxidoreductases; Physical Education and Training; Thiobarbituric Acid Reactive Substances; Vitamin A; Vitamin E

Oxidant and free radical-generating system were used to promote oxidative damage in erythrocytes. Among the oxidants used, phenylhydrazine represents one of the most investigated intracellular free radical-generating probes, which in the presence of haemoglobin auto-oxidises and give rise to hydroxyl radical, a marker for cellular damage. Erythrocyte, as a single cell, is a good model to be used for studying the haemolytic mechanism of anaemia. Our present investigations reveal increased lipid peroxidation of erythrocyte using phenylhydrazine as well as other oxygen-generating systems (hydrogen peroxide, iron with hydrogen peroxide). It has further been observed that not only lipid peroxidation, phenylhydrazine causes significant elevation in methemoglobin formation, catalase activity and turbidity, in the above system, which are the typical characteristics of haemolytic anaemia. However, exogenous administration of green tea leaf extract and ascorbic acid as natural antioxidants and free radical scavengers were shown to protect separately increased lipid peroxidation caused by phenylhydrazine, though the degree of protection is more in case of green tea leaf extract than ascorbic acid. Results suggest that oxidative damage in vivo due to haemolytic disease may be checked to some extent by using natural antioxidants.

Topics: Animals; Ascorbic Acid; Erythrocytes; Goats; Hemolysis; Lipid Peroxidation; Oxidants; Oxidative Stress; Plant Extracts; Plant Leaves; Tea

Three experiments were carried out with rats (experiments 1 and 2) and guinea pigs (experiment 3) to study the effect of oxidized fats, in interaction with dietary concentrations of vitamins E and C, on the antioxidant status of erythrocytes and the rate of haemolysis. In experiment 1, diets with fresh or thermoxidized fats, containing either 25 or 250 mg alpha-tocopherol equivalents/kg were used; experiment 2 included diets with fresh or thermoxidized fats, containing 25 mg alpha-tocopherol equivalents/kg; in experiment 3, besides a control diet with a fresh fat, diets containing thermoxidized fats with various concentrations of Vitamin E (35 vs. 175 mg alpha-tocopherol equivalent/kg) and Vitamin C (300 vs. 1000 mg/kg) were used. Rats and guinea pigs fed diets with oxidized fats had reduced concentrations of glutathione in erythrocytes as compared with animals fed the fresh fat diets. In rats fed oxidized fats, the activity of catalase and in guinea pigs fed oxidized fats, the activity of glutathione peroxidase plus the concentration of alpha-tocopherol was reduced in erythrocytes as compared with animals fed the equivalent fresh fat diets. The concentrations of alpha-tocopherol in erythrocytes were increased by both, Vitamin E and Vitamin C while the concentrations of glutathione were independent of the concentrations of those vitamins in the diets. Erythrocytes of guinea pigs fed oxidized fats also showed an increased susceptibility to haemolysis during incubation in hypotonic salt solutions; this effect could be improved by increasing the concentrations of both, Vitamin E and Vitamin C. Parameters of in vivo haemolysis (activities of lactate dehydrogenase and acidic phosphatase and concentrations of potassium and free haemoglobin in plasma) were not adversely affected in rats and guinea pigs fed the oxidized fats as compared with animals fed the fresh fats. The study shows that dietary oxidized fats reduce the antioxidant status of erythrocytes and increase their susceptibility against haemolysis but do not increase the rate of haemolysis in vivo.

Topics: alpha-Tocopherol; Animals; Antioxidants; Ascorbic Acid; Diet; Dietary Fats; Erythrocytes; Glutathione; Guinea Pigs; Hemolysis; Male; Oxidation-Reduction; Rats; Rats, Sprague-Dawley; Weight Gain

Diabetes is an oxidative stress-related disorder in which erythrocyte zinc uptake may vary as compared to healthy individuals. Since zinc is one of the important antioxidant trace metals, some functional indices of erythrocyte zinc status, ie in vitro zinc uptake, osmotic fragility and glucose uptake, were compared in Type 2 diabetic subjects (n=43) and healthy controls (n=22). The associations of these indices with plasma levels of antioxidants and micronutrients were examined. The trace metals were analyzed by atomic absorption spectrophotometer. Vitamins were estimated using spectrophotometric and spectroflourometric methods. In vitro zinc uptakes of healthy subjects were 17 to 52% higher (p<0.01) than those for diabetic subjects. The osmotic fragility for diabetic cells was 2.2 to 1.5 times higher than the healthy cells in 0.85-0.5% NaCl solutions (p<0.05). Percent hemolysis at 0.75, 0.65 and 0.55% NaCl had significant negative correlations (p<0.05) with in vitro zinc uptakes and that at 0.50% NaCl had a positive correlation with HbA1c levels (p<0.05). The in vitro zinc uptakes of erythrocytes in healthy subjects showed a strong negative correlation (p<0.01) with percent hemolysis at 0.75, 0.65 and 0.55% NaCl, a positive correlation with plasma zinc (r=0.33, p<0.05) and a strong negative correlation with plasma selenium and iron, hemoglobin and serum ceruloplasmin indicating antagonistic behavior of copper, iron and selenium with zinc uptake (p<0.01). Furthermore, erythrocyte super oxide dismutase (SOD), plasma ascorbic acid and status of riboflavin and thiamine were negatively correlated with in vitro zinc uptakes of erythrocytes in healthy subjects (p<0.01). These associations in the diabetic subjects were weaker than normal. Erythrocyte zinc uptake and osmotic fragility could be biomarkers of long-term zinc status and decrease of zinc uptake may be one of the features of diabetic patients.

Topics: Adult; Aged; Antioxidants; Ascorbic Acid; Blood Glucose; Ceruloplasmin; Diabetes Mellitus, Type 2; Erythrocytes; Female; Glycated Hemoglobin; Hemoglobins; Hemolysis; Humans; In Vitro Techniques; Iron; Male; Metals; Micronutrients; Middle Aged; Osmotic Fragility; Oxidative Stress; Riboflavin; Selenium; Sodium Chloride; Spectrometry, Fluorescence; Spectrophotometry, Atomic; Superoxide Dismutase; Thiamine; Vitamins; Zinc

Dietary polyphenolics in fruits, vegetables, wines, spices and herbal medicines have beneficial antioxidant, anti-inflammatory and anticancer effects. However, we have observed that dietary polyphenolics with phenol rings were metabolized by peroxidase to form prooxidant phenoxyl radicals which, in some cases were sufficiently reactive to cooxidize GSH or NADH accompanied by extensive oxygen uptake and reactive oxygen species formation. The order of catalytic effectiveness found for oxygen activation when polyphenolics were metabolized by peroxidase in the presence of GSH was phloretin>phloridzin>4,2'-dihydroxy chalcone>p-coumaric acid>naringenin>apigenin>curcumin>resveratrol>isoliquiritigenin>capsaicin>kaempferol. Ascorbate was also cooxidized by the phenoxyl radicals but without oxygen activation. Polyphenolics with catechol rings also cooxidized ascorbate, likely mediated by semiquinone radicals. The order of catalytic effectiveness found for ascorbate cooxidation was fisetin luteolin, quercetin, >eriodictyol, caffeic acid, nordihydroguaiaretic acid>catechin>taxifolin, catechol. NADH was stoichiometrically oxidized without oxygen uptake which, suggests that o-quinone metabolites were responsible. GSH was not cooxidized and GSH conjugates were formed, likely mediated by the o-quinone metabolites. Incubation of hepatocytes with dietary polyphenolics containing phenol rings was found to partially oxidize hepatocyte GSH to GSSG while polyphenolics with a catechol ring were found to deplete GSH through formation of GSH conjugates. Dietary polyphenolics with phenol rings also oxidized human erythrocyte oxyhemoglobin and caused erythrocyte hemolysis more readily than polyphenolics with catechol rings. It is concluded that polyphenolics containing a phenol ring are generally more prooxidant than polyphenolics containing a catechol ring.

Topics: Animals; Ascorbic Acid; Cell Survival; Diet; Erythrocytes; Flavonoids; Hemolysis; Hepatocytes; Humans; Intracellular Membranes; Male; Membrane Potentials; Mitochondria; Oxidation-Reduction; Oxidative Stress; Oxyhemoglobins; Phenols; Polymers; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species

Resveratrol and its analogs, six other polyhydroxystilbenes, were synthesized and their antioxidative activities were evaluated in vitro by determination of the levels of malondialdehyde and hydrogen peroxide. Results clearly exhibited that resveratrol and its analogs had various potencies in inhibiting lipid peroxidation in rat brain, kidney, and liver homogenates and rat erythrocyte hemolysis. Several polyhydroxystilbenes were found to be more active than resveratrol in these models, and structure-activity relationship studies on polyhydroxystilbenes are described in this paper.

Topics: Animals; Antioxidants; Ascorbic Acid; Brain Chemistry; Erythrocytes; Hemolysis; Hydrogen Peroxide; Indicators and Reagents; Kidney; Liver; Male; Malondialdehyde; Rats; Rats, Wistar; Resveratrol; Stilbenes; Structure-Activity Relationship

Hemidesmus indicus R. Br. (Asclepiadaceae) is a well known drug in Ayurveda system of medicine. In the present study, antioxidant activity of methanolic extract of H. indicus root bark was evaluated in several in vitro and ex vivo models. Further, preliminary phytochemical analysis and TLC fingerprint profile of the extract was established to characterize the extract which showed antioxidant properties. The in vitro and ex vivo antioxidant potential of root bark of H. indicus was evaluated in different systems viz. radical scavenging activity by DPPH reduction, superoxide radical scavenging activity in riboflavin/light/NBT system, nitric oxide (NO) radical scavenging activity in sodium nitroprusside/Greiss reagent system and inhibition of lipid peroxidation induced by iron-ADP-ascorbate in liver homogenate and phenylhydrazine induced haemolysis in erythrocyte membrane stabilization study. The extract was found to have different levels of antioxidant properties in the models tested. In scavenging DPPH and superoxide radicals, its activity was intense (EC50 = 18.87 and 19.9 microg/ml respectively) while in scavenging NO radical, it was moderate. It also inhibited lipid peroxidation of liver homogenate (EC50 = 43.8 microg/ml) and the haemolysis induced by phenylhydrazine (EC50 = 9.74 microg/ml) confirming the membrane stabilization activity. The free radical scavenging property may be one of the mechanisms by which this drug is effective in several free radical mediated disease conditions.

Topics: alpha-Tocopherol; Animals; Antioxidants; Apocynaceae; Ascorbic Acid; Biphenyl Compounds; Chromatography, Thin Layer; Curcumin; Erythrocyte Membrane; Free Radical Scavengers; Hemolysis; Lipid Peroxidation; Liver; Medicine, Ayurvedic; Nitric Oxide; Oxidants; Phenylhydrazines; Picrates; Plant Bark; Plant Extracts; Plant Roots; Pyrogallol; Rats; Superoxides

Prolonged low-medium intensity exercise is associated with increased oxidative stress in humans. We hypothesized that competitive equine endurance racing would induce changes in circulatory antioxidants and produce systemic oxidative stress. Forty horses competing in a 140-km endurance race in warm conditions [shade temperature 15-19 degrees C; 62-88% relative humidity (%RH)] were sampled before (Pre), immediately after exercise (End) and at approximately 16 h into recovery (+16 h). Plasma ascorbic acid concentration was not different between Pre [11.1 (median); 4.6-20.3 micromol/L (range)] and End [9.7; 3.0-38.9 (range) micromol/L] but was significantly decreased at +16 h (5.5; 2.8-15.5 micromol/L; P < 0.05). Total red cell hemolysate glutathione (TGSH) concentration was significantly reduced by exercise (Pre 1261; 883-1532 micromol/L; End 1065; 757-1334 micromol/L; P < 0.05) and at +16 h recovery (1032; 752-1362 micromol/L; P < 0.05). Glutathione redox ratio was unchanged by exercise but was significantly decreased at +16 h compared with that at both Pre and End (P < 0.05). The concentration of total barbituric acid reactive substances (TBARS) in plasma was increased compared with that at Pre (309; 66-1048 nmol/L), both at End (408; 170-1196 nmol/L; P < 0.05) and +16 h (380; 99-1161 nmol/L; P < 0.05). alpha-Tocopherol was unchanged by exercise or recovery. Mean race speed was 16.5 +/- 1.6 km/h and ranged from 13.9 to 19.7 km/h. Mean speed during competition in horses that completed the full 140 km (n = 28) was significantly correlated with end of exercise ascorbic acid (r = 0.622; P = 0.0004). Although there were increases in creatine phosphokinase (CK), aspartate aminotransferase (AST) and TBARS and a loss of TGSH, this study failed to demonstrate evidence of classical oxidative stress.

Topics: Animals; Antioxidants; Ascorbic Acid; Erythrocytes; Glutathione; Glutathione Disulfide; Hemolysis; Horses; Motor Activity; Physical Endurance; Time Factors

Antioxidative activities of the extracts from Sargassum siliquastrum were determined using the inhibition of red blood cell (RBC) hemolysis induced by 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) radicals, suppression of lipid peroxidation using rat brain homogenate, and scavenging activity of superoxide radicals. The dichloromethane fraction isolated from the methanol crude extract by differential solvent extractions exhibited the strongest antioxidant activity in both RBC hemolysis and lipid peroxidation assays. This fraction was further fractionated into four subfractions F1-F4 by silica gel column chromatography. F1 was found to be most effective in protecting RBC against AAPH radicals and in inhibiting lipid peroxidation. On the basis of thin-layer chromatography and UV and IR spectra analyses, all subfractions contained phenolic compounds. However, there was no correlation between the above antioxidant potency and total phenolic compounds estimated by using the Folin-Ciocalteau method.

Topics: Amidines; Animals; Antioxidants; Ascorbic Acid; Brain Chemistry; Chemical Fractionation; Chromatography, Thin Layer; Ferrous Compounds; Free Radicals; Hemolysis; Lipid Peroxidation; Male; Methylene Chloride; Phaeophyceae; Phenols; Rats; Rats, Sprague-Dawley; Seaweed; Spectrophotometry, Ultraviolet

This study was carried out to investigate sequel of oxidative insult to human erythrocytes induced by a water-soluble radical initiator, 2,2'-azobis-(amidinopropane) dihydrochloride (AAPH) and the effect of a commercially available mixed antioxidant (Blackmores, BioAce Excel), containing alpha-tocopherol, ascorbic acid, beta-carotene and some herbal extracts (containing grape seed catechins and milk thistle derived silybin), on lipid peroxidation, degradation of membrane proteins and haemolysis. We performed this study in order firstly to clarify aspects of the mechanism of AAPH induced free radical damage in human erythrocytes and secondly to establish in vitro conditions by which the efficacy of mixed antioxidant preparations may fairly and objectively be compared. In the process of oxidation initiated by peroxyl radical, a rapid loss of reduced glutathione occurred in the first 60 min. Formation of thiobarbitric acid-reactive substances indicative of lipid peroxidation increased subsequently and almost reached maximal levels at 180 min before significant apparent degradation of membrane proteins was detected. At this point, a significant haemolysis occurred. This sequence of events is consistent with the idea that haemolysis is a consequence of lipid peroxidation and the degradation of membrane proteins. The mixed commercial antioxidant, which suppressed lipid peroxidation and protected membrane proteins against degradation induced by peroxyl radicals, also effectively delayed AAPH induced haemolysis. The system we describe provides a sound objective basis for the in vitro comparison of the potential efficacy of the hundreds of antioxidant nutritional supplements currently available in the market place.

Topics: Amidines; Antioxidants; Ascorbic Acid; beta Carotene; Catechin; Drug Combinations; Erythrocytes; Fruit; Glutathione; Hemolysis; Humans; In Vitro Techniques; Kinetics; Lipid Peroxidation; Membrane Proteins; Methemoglobin; Oxidants; Plant Extracts; Seeds; Silymarin; Thiobarbituric Acid Reactive Substances; Vitamin E

To measure metabolic rates of the hexose monophosphate shunt (HMPS) in erythrocytes of rhinoceroses, and to test the hypothesis that low concentrations of endogenous ATP in erythrocytes impair HMPS capacity, thereby increasing susceptibility to oxidant-induced hemolysis.. 13 black and 3 white rhinoceroses, free-ranging in several regions of southern Africa, and 1 Sumatran rhinoceros in US captivity.. HMPS fluxes were measured in rhinoceros erythrocytes with carbon-labeled glucose in the presence and absence of known HMPS activators.. Compared with values for human erythrocytes, mean basal state HMPS fluxes were appreciably lower (22 to 46%) in all 3 rhinoceros species studied. Shunt activators increased HMPS rates approximately 5-fold over basal rates in rhinoceros erythrocytes, compared with increases in humans of 10-fold with ascorbate and 15-fold with methylene blue. Stimulated HMPS rates in human erythrocytes were quantitatively 5- to 10-times greater than those observed in rhinoceros erythrocytes. Overall HMPS catabolic rates were completely independent of intracellular ATP concentrations.. HMPS glycolytic and recycling rates and responses to activators are inherently low in erythrocytes from 3 species of rhinoceros, likely contributing to (but not solely responsible for) the high susceptibility of black rhinoceroses to oxidant-induced hemolysis. Slow erythrocyte HMPS capacities were independent of intracellular ATP concentrations, invalidating a current hypothesis regarding the pathogenesis of hemolytic anemia in captive black rhinoceroses. Limitations in HMPS capacities emphasize the importance of protecting rhinoceroses from exposure to drugs, chemicals, toxins, foodstuffs, and other conditions known to increase production of oxidizing metabolites, reactive oxygen species, and free radicals.

Topics: Adenosine Triphosphate; Animals; Ascorbic Acid; Blood Glucose; Enzyme Inhibitors; Erythrocytes; Hemolysis; Humans; Indonesia; Mammals; Methylene Blue; Pentose Phosphate Pathway; Perissodactyla; Radiometry; South Africa

Oxidant stress is one of the factors proposed to be responsible for damaged erythrocytes observed during and after exercise. The impact of exertional oxidant stress after acute exhaustive treadmill running on erythrocyte damage was investigated in sedentary (Sed) and exercise-trained (ET) rats treated with or without antioxidant vitamins C and E. Exhaustive exercise led to statistically significant increments in the levels of thiobarbituric acid-reactive substance (TBARS) and H2O2-induced TBARS in Sed rats and resulted in functional and structural alterations in erythrocytes (plasma hemoglobin concentrations, methemoglobin levels, and rise in osmotic fragility of erythrocytes with decrease in erythrocyte deformability). Administration of antioxidant vitamin for 1 mo before exhaustive exercises prevented lipid peroxidation (TBARS, H2O2-induced TBARS) in Sed rats without any functional or structural alterations in erythrocytes. Parameters indicating erythrocyte lipid peroxidation and deterioration after exhaustive exercise in rats trained regularly with treadmill running for 1 mo were not different from those in Sed controls. Erythrocyte lipid peroxidation (TBARS) increased in exhausted-ET rats compared with ET controls; however, the plasma hemoglobin, methemoglobin levels, and erythrocyte osmotic fragility and deformability did not differ. Exhaustive exercise-induced lipid peroxidation in ET rats on antioxidant vitamin treatment was prevented, whereas functional and structural parameters of erythrocytes were not different from those of the ET controls. We conclude that exertional oxidant stress contributed to erythrocyte deterioration due to exercise in Sed but not in ET rats.

Topics: Animals; Antioxidants; Ascorbic Acid; Body Weight; Erythrocyte Deformability; Erythrocytes; Hemolysis; Hydrogen Peroxide; Male; Methemoglobin; Organ Size; Osmotic Fragility; Oxidants; Oxidative Stress; Physical Conditioning, Animal; Physical Exertion; Rats; Rats, Wistar; Thiobarbituric Acid Reactive Substances; Vitamin E

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

Topics: Adult; Aged; Ascorbic Acid; Catalase; Erythrocytes; Female; Ferritins; Glutathione; Glutathione Peroxidase; Hemoglobins; Hemolysis; Humans; Male; Malondialdehyde; Middle Aged; Oxidative Stress; Renal Dialysis; Superoxide Dismutase; Vitamin E

Ascorbic acid, or vitamin C, has been reported to lower erythrocyte sorbitol concentrations, and present studies were performed to determine the mechanism of this effect. Incubation of erythrocytes with increasing concentrations of glucose (5-40 mM) progressively increased erythrocyte sorbitol contents, reflecting increased flux through aldose reductase. At extracellular concentrations of 90 microM, both ascorbic acid and its oxidized form, dehydroascorbate, decreased intracellular sorbitol by 25 and 45%, respectively. This inhibition was not dependent on the extracellular glucose concentration, or on erythrocyte contents of free NADPH or GSH. To test for a direct effect of ascorbate on aldose reductase, erythrocyte hemolysates were prepared and supplemented with 100 microM NADPH. Hemolysates reduced glucose to sorbitol in a dose-dependent manner that was inhibited with a Ki of 120 microM by the aldose reductase inhibitor tetramethylene glutaric acid. Above 100 microM, ascorbic acid also lowered hemolysate sorbitol generation by about 30%. Studies with ascorbic acid derivatives showed that the reducing capacity of ascorbic acid was not required for inhibition of sorbitol production from glucose in erythrocyte hemolysates. These results show that high, but physiologic, concentrations of ascorbic acid can directly inhibit erythrocyte aldose reductase, and provide a rationale for the use of oral vitamin C supplements in diabetes.

Topics: Aldehyde Reductase; Ascorbic Acid; Dehydroascorbic Acid; Enzyme Inhibitors; Erythrocytes; Glucose; Hemolysis; Humans; Intracellular Membranes; NADP; Sorbitol

Human erythrocytes efficiently reduce dehydroascorbic acid (DHA) to ascorbate, which helps to maintain the ascorbate content of blood. Whereas erythrocyte DHA reduction is thought to occur primarily through a direct chemical reaction with GSH, this work addresses the role of enzyme-mediated DHA reduction by these cells. The ability of intact erythrocytes to recycle DHA to ascorbate, estimated as DHA-dependent ferricyanide reduction, was decreased in parallel with GSH depletion by glutathione-S-transferase substrates. In contrast, the sulfhydryl reagent phenylarsine oxide inhibited DHA reduction to a much greater extent than it decreased GSH in intact cells. DHA reduction in excess of that due to a direct chemical reaction with GSH was also observed in freshly prepared hemolysates. Hemolysates likewise showed NADPH-dependent reduction of DHA that appeared due to thioredoxin reductase, because this activity was inhibited 68% by 10 microM aurothioglucose, doubled by 5 microM E. coli thioredoxin, and had an apparent Km for DHA (1.5 mM) similar to that of purified thioredoxin reductase. Additionally, aurothioglucose-sensitive, NADPH-dependent DHA reductase activity was decreased 80% in hemolysates prepared from phenylarsine oxide-treated cells. GSH-dependent DHA reduction in hemolysates was more than 10-fold that of NADPH-dependent reduction. Nonetheless, the ability of phenylarsine oxide to decrease DHA reduction in intact cells with little effect on GSH suggests that enzymes, such as thioredoxin reductase, may contribute more to this activity than previously considered.

Topics: Arsenicals; Ascorbic Acid; Aurothioglucose; Cell Membrane; Cell-Free System; Dehydroascorbic Acid; Dinitrochlorobenzene; Dose-Response Relationship, Drug; Enzymes; Erythrocytes; Ferricyanides; Glutathione; Glutathione Transferase; Hemolysis; Humans; Ketones; NADP; Thioredoxin-Disulfide Reductase; Vitamin E

Exposure of intact rabbit erythrocytes or erythrocyte lysates to ascorbic acid/FeCl3 in a glucose-free saline promoted a rapid decline in reduced glutathione and this response was paralleled by inactivation of hexokinase. Under the same conditions, the activity of the enzymes glyceraldehyde-3-phosphate dehydrogenase and glucose-6-phosphate dehydrogenase did not show appreciablevariations in intact cells, but was severely inhibited in the cell-free system. Similar results were obtained by replacing ascorbic acid/FeCl3 with dehydroascorbic acid. In addition, both treatments effectively inhibited the activity of purified hexokinase as well as those of glucose-6-phosphate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase. Further studies using the cell-free system indicated that the inhibition of enzyme activities elicited by either of the two treatments was effectively counteracted by the specific substrates of these enzymes. The fact that the hexokinase substrate glucose freely permeates the plasma membrane, unlike the substrates of glucose-6-phosphate dehydrogenase and glyceraldehyde-3-phosphatedehydrogenase, explains the selective inhibition of hexokinase observed in intact cells. The above results also indicate that dehydroascorbic acid is an inhibitor of these enzymes and strongly suggest that it is at least in part responsible for the effects mediated by the cocktail ascorbic acid/FeCl3.

Topics: Animals; Ascorbic Acid; Chlorides; Dehydroascorbic Acid; Enzyme Activation; Erythrocytes; Ferric Compounds; Glucosephosphate Dehydrogenase; Glutathione; Glyceraldehyde-3-Phosphate Dehydrogenases; Hemolysis; Hexokinase; Rabbits; Substrate Specificity

A two way regression study was performed to investigate the interactions between vitamins C and E, and the influence of dietary vitamin C on the development of vitamin E deficiency in first feeding Atlantic salmon. The fish were fed three levels of all-rac-alpha-tocopheryl acetate (0, 150, and 300 mg/kg), each with six levels of ascorbate monophosphate (0, 7.5, 15, 30, 45, and 60 mg/kg ascorbic acid equivalents). Vitamin C protected the fish against vitamin E deficiency in a dose dependent manner, as seen from the data on growth, mortality, hematology, and lipid oxidation in the liver, indicated by the concentration of malondialdehyde. Vitamin C did not influence the tissue levels of vitamin E, except in vitamin C deficiency, which induced a large drop in liver vitamin E concentration. The liver level of vitamin C increased in response to supplementation of both vitamins. The results indicate two different interaction mechanisms: a synergistic effect of simultaneous protection of the water and lipid phases against oxidation, and regeneration of vitamin E from the vitamin E radical by ascorbic acid.

Topics: Animals; Ascorbic Acid; Diet; Glutathione; Hemolysis; Lipid Metabolism; Liver; Malondialdehyde; Oxidation-Reduction; Regression Analysis; Salmon; Vitamin E Deficiency; Weight Gain

The present study was designed to investigate the effect of antioxidant supplementation on the in vitro osmotic fragility of erythrocytes from zinc-deficient rats. Rats were fed either a zinc-adequate diet, zinc-deficient diet or a zinc-deficient diet enriched either with vitamin C or vitamin E or beta-carotene. Components of the primary antioxidant system of erythrocytes, parameters of hemolysis in vivo and indicators of liver injuries were also examined. In order to ensure adequate and identical food intake rats were force-fed by intragastric tube. The supplementation with antioxidants led to a marked improvement of the osmotic fragility without having influenced zinc status of the animals and components of the antioxidant system. The strongest effect was exerted by vitamin E. The rats fed the zinc-adequate diet (control group) showed unusually high values of erythrocytes osmotic fragility. Therefore there was no difference between control group and zinc-deficient group. A possible reason for this is discussed. Zinc deficiency led to a reduction of serum zinc concentration and alkaline phosphatase activity as well as to changes in the antioxidant system of erythrocytes characterized by a decrease of glutathione and an increase of glutathione S-transferase activity. Superoxide dismutase activity in serum decreased. There was no indication for hemolysis in vivo and for liver injuries.

Topics: Alkaline Phosphatase; Analysis of Variance; Animals; Antioxidants; Ascorbic Acid; beta Carotene; Eating; Erythrocytes; Food, Fortified; Glutathione Transferase; Hemolysis; Male; Models, Statistical; Osmotic Fragility; Rats; Rats, Sprague-Dawley; Superoxide Dismutase; Vitamin E; Zinc

We have isolated from bovine brain a protein with a high capacity to inhibit the copper ion-catalyzed oxidation of L-ascorbate and identified it as S100b protein, an EF-hand calcium-binding protein, by sequencing its proteolytic peptides. Copper binding studies showed that this protein has four copper-binding sites per dimeric protein molecule with a dissociation constant of 0.46 microM and that in the presence of L-ascorbate, copper ions bind to a total of six binding sites with a great increase in affinity. Furthermore, we examined whether S100b protein can prevent copper-induced cell damage. Bovine S100b protein was found to suppress dose-dependently the hemolysis of mouse erythrocytes induced by CuCl2. We transformed Escherichia coli cells with pGEX-5X-3 vector containing a cDNA for rat S100b protein, so that this protein could be expressed as a fusion protein with glutathione S-transferase. The transformed cells were demonstrated to be markedly resistant to a treatment with CuCl2 plus H2O2 as compared with the control cells expressing glutathione S-transferase alone. These results indicate that S100b protein does suppress oxidative cell damage by sequestering copper ions.

Topics: Amino Acid Sequence; Animals; Ascorbic Acid; Calcium-Binding Proteins; Cattle; Copper; Dose-Response Relationship, Drug; Drug Resistance; Escherichia coli; Hemolysis; Hydrogen Peroxide; Molecular Sequence Data; Nerve Growth Factors; Oxidation-Reduction; Oxidative Stress; Rats; Recombinant Fusion Proteins; S100 Calcium Binding Protein beta Subunit; S100 Proteins

Captopril ([2S]-1-[3-mercapto-2-methyl-propionyl]-L-proline) was found to protect erythrocytes from hemolysis caused by 2,2'-azobis (2-amidinopropane) (AAPH) and hypochlorite, erythrocyte membranes from lipid peroxidation caused by tert-butyl hydroperoxide (tBOOH) and hypochlorite, erythrocyte membrane ATPases from inactivation caused by tBOOH and hemoglobin from oxidation caused by AAPH and tBOOH. In all these systems enalapril ([S]-1-[N-(1-[ethoxycarbonyl]-3-phenylpropyl)-L-alanyl]-L-proline) was not protective or even increased the damage, especially with hypochlorite, probably due to chloramine formation. Captopril but not enalapril inhibited ascorbate autoxidation caused by Cu2+, which indicates that captopril binds Cu2+. On the other hand, deoxyribose degradation caused by iron and copper ions and DNA damage by o-phenanthroline/Cu2+/H2O2/beta-mercaptoethanol was enhanced by both captopril and enalapril. The effect of captopril was usually higher, apparently due to the reducing properties of captopril, which could reduce metal ions enabling their participation in the Fenton reaction. These results indicate that only -SH-group-containing inhibitors of angiotensin-converting enzyme (ACE) may exhibit antioxidant properties, and that the antioxidant/prooxidant action of ACE inhibitors depends on the system studied in vitro.

Topics: Adenosine Triphosphatases; Angiotensin-Converting Enzyme Inhibitors; Antioxidants; Ascorbic Acid; Captopril; Deoxyribose; DNA Damage; Enalapril; Erythrocyte Membrane; Hemolysis; Humans; Lipid Peroxidation; Oxidation-Reduction

The sulphonamide-derived oral antidiabetic drugs carbutamide, chlorpropamide, glibenclamide, glibornuride, gliclazide, glipizide, gliquidone, glisoxepide, glymidine, tolazamide and tolbutamide were investigated for photohemolytic properties in vitro. Irradiation with a SOL 3 apparatus (solar simulating irradiation) revealed hemolysis in the presence of chlorpropamide, glipizide, gliquidone, glymidine and tolbutamide (all in the concentration 10(-3) mol/l). Except for glymidine, which exerted photohemolysis in the concentration 10(-4) mol/l, no hemolytic effects were seen in the concentration of 10(-4) mol/l or 10(-5) mol/l. Irradiation with TL 12 light bulbs (UVB), a UVASUN 5000 apparatus (UVA) or an experimental lamp (visible light) did not induce phototoxic hemolysis with either of the test substances. Addition of the antioxidants ascorbic acid, alpha-tocopherol or superoxide dismutase significantly inhibited the phototoxic hemolysis. Investigations carried out in a nitrogen-rich atmosphere reduced the hemolysis as well. These findings indicate an involvement of reactive oxygen species in the mechanism of action of the hemolytic process in the presence of oral antidiabetic drugs.

Topics: Antioxidants; Ascorbic Acid; Carbutamide; Chlorpropamide; Dose-Response Relationship, Drug; Gliclazide; Glipizide; Glyburide; Hemolysis; Humans; Hypoglycemic Agents; Nitrogen; Radiation-Sensitizing Agents; Reactive Oxygen Species; Sulfonamides; Sulfonylurea Compounds; Superoxide Dismutase; Tolazamide; Tolbutamide; Ultraviolet Rays; Vitamin E

Hemolysis of human erythrocytes induced by free radicals initiated from water-soluble, 2,2'-azobis(amidinopropane) dihydrochloride (AAPH) has been investigated. The formation of the radical detected as DMPO (5,5-dimethyl-1-pyrroline N-oxide) adduct depended on temperature and AAPH concentration in a similar manner as hemolysis. The curve for the formation of DMPO--radical adduct, however, did not correspond directly to the hemolysis curve. The product of thiobarbituric acid-reactive materials, which reflect the extent of lipid peroxidation, could not be related directly to the hemolysis curve, too. During the hemolysis, the fluidity of the erythrocyte membrane did not change in appearance. To study whether band 3 proteins participate in the hemolysis or not, eosin-5-maleimide (EMI)-labeled ghosts were incubated in the presence of AAPH. High molecular weight band 3 was formed, and the induced circular dichrosim spectrum of the bound EMI was changed, indicating a conformational change of band 3. It was observed that ascorbic acid suppressed the hemolysis and the oxidation of band 3 dose dependently to produce an induction period. This result shows that specifically blocking band 3 oxidation inhibits the hemolysis, despite lipid peroxidation. Further, it was observed that the EMI-labeled erythrocytes revealed distinct clusters by incubation with AAPH. This means a redistribution of band 3 proteins to form hemolytic holes in the membrane. However, the time course of the conformational change of band 3 during the redistribution was not also correspondent to the hemolysis curve. These results indicate that either lipid peroxidation or redistribution of oxidized band 3 is not attributed only by itself to the hemolysis. Thus, the hemolysis was interpreted by a simple competitive reaction model between lipid peroxidation and redistribution of oxidized band 3. This model explained well the hemolysis curves.

Topics: Amidines; Anion Exchange Protein 1, Erythrocyte; Ascorbic Acid; Circular Dichroism; Cyclic N-Oxides; Electron Spin Resonance Spectroscopy; Erythrocytes; Free Radicals; Hemolysis; Humans; In Vitro Techniques; Kinetics; Lipid Peroxidation; Membrane Lipids; Oxidation-Reduction; Protein Conformation; Solubility; Spin Labels; Thermodynamics; Water

Substantial evidence supports the theory that free radicals, especially oxygen radicals, are involved in the process of aging. The human organisms have two ways to fight them: an enzymatic way with enzymatic intervention like superoxide dismutase, catalase... and a chemical way with the intervention of scavengers such as vitamins, cysteine, methionine, gluthatione... The aim of this work was to determine that an intakes of vitamins association: vitamin E, vitamin C and beta carotene induce an increase of singlet oxygen protection of erythrocytes' subjects. The method was based on the haemolytic effect of singlet oxygen which is generated by irradiation of hematoporphyrine at 365 nm, in 22 p. cent suspension of erythrocytes' subjects. Results show that a supply of beta carotene (15 or 30 mg/day), vitamin E (15 mg/day) and vitamin C (30 mg/day) involves an increase of singlet oxygen protection of erythrocytes of subjects. This protection appears very quickly after 15 days of treatment.

Topics: Administration, Oral; Ascorbic Acid; beta Carotene; Carotenoids; Erythrocyte Aging; Erythrocytes; Hematoporphyrins; Hemolysis; Humans; Oxygen; Photochemistry; Reference Values; Singlet Oxygen; Ultraviolet Rays; Vitamin E

Topics: Antioxidants; Ascorbic Acid; Butylated Hydroxytoluene; Chromium; Erythrocytes; Hemoglobins; Hemolysis; Humans; Kinetics; Maleates; Oxidation-Reduction; Oxyhemoglobins; Salicylates; Salicylic Acid; Vitamin E

Topics: Adult; Anemia, Hemolytic; Ascorbic Acid; Carbonated Beverages; Erythrocytes; Hemoglobinuria, Paroxysmal; Hemolysis; Humans; Male

A detailed investigation on the anti-inflammatory activity of the butanol fraction of a methanol extract (BMEL) of the defatted leaves of Paederia foetida was undertaken to find the pharmacological basis for the ethnomedical use of the plant. This fraction produced a significant inhibition of granulation tissue formation in cotton-pellet implanted rats. It decreased liver aspartate transaminase activity without affecting serum aspartate transaminase activity. It did not, however, affect adrenal weight and ascorbic acid content significantly, thus ruling out a stimulation of the adrenal-pituitary axis. BMEL antagonised hyposaline-induced haemolysis of human red blood cells and an elevation of rat serum acid phosphatase activity, indicating the presence of a membrane stabilising activity. It also inhibited the elevation of serum orosomucoid levels in rats, suggesting the possibility of the presence of disease-modifying antirheumatic activity. The results indicate that there is some rationale behind the ethnomedical use of the plant for treating inflammatory disorders.

Topics: 1-Butanol; Acid Phosphatase; Adrenal Glands; Animals; Anti-Inflammatory Agents; Ascorbic Acid; Aspartate Aminotransferases; Blood Proteins; Butanols; Chemical Fractionation; Disease Models, Animal; Erythrocyte Membrane; Female; Granuloma; Hemolysis; Humans; Liver; Male; Medicine, Traditional; Methanol; Organ Size; Orosomucoid; Plant Extracts; Rats; Spleen

Topics: Aged; Ascorbic Acid; Hemoglobinuria, Paroxysmal; Hemolysis; Humans; Male; Tea

The azo initiator of peroxyl radicals 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) induces oxidative hemolysis in human erythrocytes and subsequent hemoglobin oxidation. Using the degree of hemolysis versus time as an indication of the oxidative damage it was found that i) both reduced and oxidized alpha-lipoic acid protected against oxidative damage; ii) simultaneous treatment of erythrocytes with ascorbate and dihydrolipoate or alpha-lipoate has a synergistic tendency to protect cells against hemolysis; iii) glutathione in combination with dihydrolipoic acid or alpha-lipoic acid has an additive effect on hemolysis protection. The spin trapping reagent 5,5-dimethyl-1-pyrroline N-oxide (DMPO) formed an adduct with the peroxyl/alkoxyl radicals produced by thermal decomposition of AAPH in the presence of oxygen. The formation of this adduct was prevented by reduced or oxidized lipoic acid, reduced glutathione or ascorbate. It is concluded that AAPH-peroxyl radicals progressively damage the cells and the released hemoglobin is subsequently oxidized to methemoglobin which might further enhance the oxidative damage. The protective effect of antioxidants is exerted outside the cells by directly scavenging AAPH-alkoxyl radicals.

Topics: Amidines; Ascorbic Acid; Cyclic N-Oxides; Drug Synergism; Electron Spin Resonance Spectroscopy; Erythrocytes; Free Radicals; Glutathione; Hemolysis; Humans; Methemoglobin; Oxidation-Reduction; Stereoisomerism; Structure-Activity Relationship; Thioctic Acid

Topics: 1,2-Dipalmitoylphosphatidylcholine; Amidines; Animals; Antioxidants; Ascorbic Acid; Erythrocytes; Free Radicals; Hemolysis; In Vitro Techniques; Mice; Solubility

It was previously shown that the reductive debromination of BrCCl3 to trichloromethyl radical by human hemoglobin leads to formation of dissociable altered heme products, two of which are identical to those formed from myoglobin and one which is novel. In this study, we have elucidated the structure of this novel adduct with the use of mass spectrometry, as well as 1H and 13C NMR as a substitution product of a -C(Cl) = CCl2 moiety for a beta-hydrogen atom on the prosthetic heme's ring I vinyl group. From studies with the use of 13C-enriched BrCCl3, it was determined that the added carbon atoms were derived from 2 eq of BrCCl3. A mechanism that involves multiple reductive events and a radical cation heme intermediate is proposed. Consistent with this mechanism, cellular reductants were found to selectively enhance the amount of this novel dissociable heme adduct. These studies reveal fine differences between myoglobin and hemoglobin in the accessibility of reactive intermediates to the ring I vinyl group, as well as the potential importance of cellular reductants on the course of heme alteration.

Topics: Ascorbic Acid; Binding Sites; Bromotrichloromethane; Carbon Isotopes; Erythrocytes; Glutathione; Heme; Hemoglobins; Hemolysis; Humans; Kinetics; Magnetic Resonance Spectroscopy; Molecular Structure; Myoglobin

The human gene-engineering gamma-interferon agent gammaferon (GF) is demonstrated to lower the level of chromosomal aberrations and release of cyclophosphamide-induced micronuclei in murine bone marrow cells. A model is suggested for regulating the ratio of single to multiple chromosomal aberrations induced by cyclophosphamide by altering the antioxidative status of the body. With this model, it was shown that the antimutagenic effects of GF were caused by two factors: the antioxidative effect of the corresponding placebo and the unknown (but none antioxidative) effect of gamma-interferon itself. Erythrocytic resistance to in vitro oxidative disturbance in the Fe2+/system was used as an integral measure of the body's antioxidative status.

Topics: Animals; Antimutagenic Agents; Ascorbic Acid; Bone Marrow; Bone Marrow Cells; Chi-Square Distribution; Chromosome Aberrations; Cyclophosphamide; Hemolysis; Interferon-gamma; Mice; Mice, Inbred C57BL; Mice, Inbred CBA; Recombinant Proteins; Species Specificity

Intraerythrocytic malaria parasites ingest the cytosol of their host cell and digest it inside their acid food vacuoles. Acidified (pH 4-5.5, 37 degrees C) human red blood cell lysates were used to simulate this process, measuring the denaturation of hemoglobin (Hb) and the release of iron, in the absence or presence of exogenous protease. Spontaneous Hb denaturation and appearance of non-heme iron were observed upon lysate acidification, its rate decreasing with increasing pH, and increasing in presence of protease. Although the pH- and proteolysis-dependent release of iron paralleled Hb denaturation, released iron accounted for only a few percent of degraded Hb. Superoxide dismutase, catalase, and various scavengers of oxidative radicals had no effect on either process, consistent with the involvement of Fe(IV) intermediates in iron release from heme. Histidine and imidazole inhibited iron release, probably by binding directly to heme. Ascorbate enhanced iron release considerably but marginally enhanced the denaturation of Hb, suggesting that redox cycling of lysate free iron accelerated further release from heme. These processes could account for the endogenous supply of iron to the malarial parasite.

Topics: Ascorbic Acid; Catalase; Cathepsin D; Cytosol; Dose-Response Relationship, Drug; Endopeptidases; Erythrocytes; Free Radical Scavengers; Hemoglobins; Hemolysis; Humans; Hydrogen-Ion Concentration; Iron; Malaria; Protease Inhibitors; Superoxide Dismutase; Time Factors

Topics: Acute Disease; Adult; Ascorbic Acid; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Male

The rate of photohaemolysis of human red blood cells sensitized by chloroaluminium phthalocyanine sulphonate is increased by ascorbate, with or without added FeCl3. Stimulation of haemolysis by ascorbate without addition of metal salt, and in the presence of a strong chelator such as desferrioxamine, is an unexpected phenomenon. Lysis rate and ascorbate concentration were directly related, suggesting that ascorbate acts as a reactant and not as a catalyst. The process also requires oxygen; azide and D2O tests indicate some participation of singlet oxygen, although to a lesser extent than in the photosensitized haemolysis in the absence of ascorbate. Kinetic considerations suggest a reaction path initiated by excited sensitizer and ascorbate, parallel to the singlet oxygen-mediated process. Because of the ubiquitous presence of ascorbate in human tissues in concentrations comparable to those of dissolved oxygen, it is quite possible that in photodynamic therapy a fraction of the photodynamic damage proceeds via a Type I, ascorbate-assisted, mechanism.

Topics: Ascorbic Acid; Chlorides; Drug Synergism; Erythrocytes; Ferric Compounds; Hemolysis; Humans; Indoles; Organometallic Compounds; Radiation-Sensitizing Agents

The Glucocard (Kyoto Daiichi Kagaku) blood glucose meter is designed for self-monitoring of blood glucose concentrations in capillary blood through use of an electrochemical test strip. Evaluated in this laboratory, the Glucocard had CVs of 4.6%, 6.6%, and 3.5% at blood glucose concentrations of 2.4, 4.1, and 18.9 mmol/L, respectively. The meter's response varied linearly with blood glucose concentration between 2.2 and 27.8 mmol/L. Hemolysis, urate, ascorbate, and acetaminophen interfered by > 5%. Different hematocrits, in the range 0.20-0.70, did not affect the measured glucose concentration. Comparison with glucose results measured in whole blood with a NOVA Stat Profile 5 instrument yielded the following: Glucocard = 0.898 NOVA--0.184 (r = 0.995). The main advantages of the Glucocard are its small sample volume (5 microL), wide linear range, and fully automated sample-handling steps, which reduce user-related variability.

Topics: Acetaminophen; Ascorbic Acid; Blood Glucose; Blood Glucose Self-Monitoring; Capillaries; Electrochemistry; Hemolysis; Humans; Quality Control; Uric Acid

A significant decrease in blood haemoglobin, reduced glutathione and protein in lung and liver, without any change in blood reduced glutathione, was observed in rats exposed to 80% oxygen. Hydrogen peroxide induced erythrocyte haemolysis was significantly increased following exposure to hyperoxia. The lungs of rats exposed to hyperoxia showed perivascular edema. Simultaneous treatment with antioxidants, vitamin A, C, or E, protected the animals against oxygen toxicity.

Topics: Animals; Antioxidants; Ascorbic Acid; Glutathione; Hemoglobins; Hemolysis; Hydrogen Peroxide; Liver; Lung; Male; Oxygen; Proteins; Rats; Rats, Inbred Strains; Vitamin A; Vitamin E

This study looked at whether oral vitamin C and vitamin E would protect the erythrocyte from oxidant damage caused by dapsone in patients with dermatitis herpetiformis.. Fifteen consecutive patients with dermatitis herpetiformis taking dapsone therapy received, in addition, 800 U/d of vitamin E for 4 weeks; then 1000 mg of vitamin C per day for 4 weeks, and, finally, combined vitamin E and vitamin C therapy for 4 weeks. Hemolysis indexes were assessed at baseline and after each 4-week period.. Statistical analysis of the results suggests that oral administration of 800 units of vitamin E daily for 4 weeks confers partial protective effect against dapsone-induced hemolysis in patients with dermatitis herpetiformis.. Partial protection against dapsone-induced hemolysis by orally administered vitamin E, if confirmed as being clinically relevant by further trials, may allow clinicians to continue dapsone therapy orally in patients who develop significant hemolysis. Prophylactic vitamin E to minimize potential hemolysis at the initiation of dapsone therapy may also be appropriate.

Topics: Adult; Ascorbic Acid; Blood Cell Count; Dapsone; Dermatitis Herpetiformis; Female; Heinz Bodies; Hemoglobins; Hemolysis; Humans; Male; Methemoglobin; Middle Aged; Reticulocytes; Vitamin E

The photohaemolytic potentials of the quinolones oxolinic acid, pipemidic acid, rosoxacin, norfloxacin, ciprofloxacin and M-193324 (synthesis intermediary) were evaluated and compared with the photohaemolysis induced by nalidixic acid. Quinolones with a piperazine group in position 7 (pipemidic acid, norfloxacin and ciprofloxacin) did not induce photohaemolysis. However, oxolinic acid, rosoxacin and M-193324 produced a concentration- and oxygen-dependent photohaemolysis. Ascorbic acid, histidine and thiourea inhibited the photohaemolysis induced by oxolinic acid, rosoxacin and M-193324, suggesting a photodynamic mechanism similar to that found with nalidixic acid. In addition, deuterium oxide increased the photohaemolysis induced by photohaemolytic quinolones, indicating that this process is mediated by singlet oxygen.

Topics: Ascorbic Acid; Hemolysis; Histidine; Humans; Light; Photolysis; Quinolones; Radiation-Sensitizing Agents; Structure-Activity Relationship; Thiourea

Dioleoyl phosphatidylcholine (PC) liposomes were ozonized and the ozonized liposomes were tested for their lytic potency on human red blood cells (RBC). Ozonation of PC liposomes generated approximately 1 mole equivalent of hydrogen peroxide (H2O2) and 2 mole equivalents of aldehydes, based on the moles of ozone consumed. The time necessary for 50% hemolysis induced by ozonized liposomes (a convenient measure of hemolytic activity) was found to depend on the extent of ozonation of the PC liposomes, indicating the formation and accumulation of hemolytic agents during ozonation. Hemolysis was also observed when RBC were incubated with nonanal, the expected product of the ozonation of oleic acid, the principle unsaturated fatty acid in the liposomes. Hydrogen peroxide, another product of PC ozonation, did not induce hemolysis; however, a combination of H2O2 and nonanal was significantly more hemolytic than nonanal alone. A ratio of 1:2 H2O2/nonanal (the ratio observed in the ozonized liposomes) provided hemolytic activity comparable to that observed with ozonized dioleoyl PC. Among different antioxidants tested, ascorbate, catalase, and glutathione peroxidase partially inhibited hemolysis induced by ozonized liposomes and by H2O2/nonanal mixtures, but they were not protective against the nonanal-induced hemolysis. Identification of H2O2 and aldehydes as cytotoxic chemical species generated from the ozonation of unsaturated fatty acids may have an important bearing on the in vivo toxicity of ozone on the lung as well as on extrapulmonary tissues.

Topics: Aldehydes; Antioxidants; Ascorbic Acid; Catalase; Erythrocyte Membrane; Hemolysis; Humans; Hydrogen Peroxide; In Vitro Techniques; Lipid Peroxides; Liposomes; Ozone; Phosphatidylcholines; Vitamin E

Topics: Ascorbic Acid; Carbonated Beverages; Child; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Male; Time Factors

Topics: Ascorbic Acid; Hemolysis; Histidine; Humans; In Vitro Techniques; Nalidixic Acid; Naphthyridines; Photolysis; Thiourea

The effects of varying the method of preparation of haemolysates on the measurement of red cell folate concentration were investigated using the Simultrac kit. The concentration of ascorbic acid did not have any significant effect on the assayed concentrations of folate, but lower concentrations were obtained when the incubation time was increased. Folate was stable for 14 days in cells when they were stored at 4 degrees C and for seven days at -25 degrees C, but instability was increased by storage in ascorbic acid, by the use of stored (4 degrees C) ascorbic acid, and by preparing the haemolysates by freeze-thaw cycling. It is recommended that haemolysates should be diluted in fresh ascorbic acid, at a concentration of 10 g/l, incubated for 60 minutes in the dark and stored at -20 degrees/25 degrees C before being assayed.

Topics: Ascorbic Acid; Blood Preservation; Edetic Acid; Erythrocytes; Folic Acid; Freezing; Hemolysis; Humans; Methods; Reagent Kits, Diagnostic; Time Factors

Cigarette smoke has been found to induce the hemolysis of rabbit erythrocytes. The particulate phase had more profound effect than the gas phase. Neither free radical scavengers such as ascorbic acid, uric acid and water-soluble vitamin E analogue nor antioxidant enzymes such as catalase and superoxide dismutase suppressed the cigarette smoke-induced hemolysis, suggesting that free radicals, hydrogen peroxide, and superoxide were not the active species.

Topics: Animals; Ascorbic Acid; Aspartate Aminotransferases; Catalase; Erythrocytes; Free Radicals; Hemoglobins; Hemolysis; Nicotiana; Plants, Toxic; Potassium; Rabbits; Smoke; Superoxide Dismutase; Uric Acid; Vitamin E

The oxidative hemolysis of rabbit erythrocytes induced by free radicals and its inhibition by chain-breaking antioxidants have been studied. The free radicals were generated from either a water-soluble or a lipid-soluble azo compound which, upon its thermal decomposition, gave carbon radicals that reacted with oxygen immediately to give peroxyl radicals. The radicals generated in the aqueous phase from a water-soluble azo compound induced hemolysis in air, but little hemolysis was observed in the absence of oxygen. Water-soluble chain-breaking antioxidants, such as ascorbic acid, uric acid, and water-soluble chromanol, suppressed the hemolysis dose dependently. Vitamin E in the erythrocyte membranes was also effective in suppressing the hemolysis. 2,2,5,7,8-Pentamethyl-6-chromanol, a vitamin E analogue without phytyl side chain, incorporated into dimyristoylphosphatidylcholine liposomes, suppressed the above hemolysis, but alpha-tocopherol did not suppress the hemolysis. Soybean phosphatidylcholine liposomes also induced hemolysis, and a lipid-soluble azo initiator incorporated into the soybean phosphatidylcholine liposomes accelerated the hemolysis. The chain-breaking antioxidants incorporated into the liposomes were also effective in suppressing this hemolysis.

Topics: Animals; Antioxidants; Ascorbic Acid; Erythrocytes; Free Radicals; Hemolysis; In Vitro Techniques; Liposomes; Oxidation-Reduction; Phosphatidylcholines; Rabbits; Reference Values; Vitamin E Deficiency

Topics: Ascorbic Acid; Erythrocytes; Flavonoids; Hematoporphyrins; Hemolysis; Humans; Photochemistry; Quercetin

Topics: Amidines; Antioxidants; Ascorbic Acid; Bilirubin; Erythrocytes; Free Radicals; Hemoglobins; Hemolysis; Humans; Potassium; Sulfhydryl Compounds; Uric Acid; Vitamin E

An excess of copper is the cause of hemolysis in a number of clinical conditions. Incubation of human erythrocyte (RBC) suspensions with copper (II) causes the formation of methemoglobin, lipid peroxidation and hemolysis. A new variant of the thiobarbituric acid (TBA) method, which minimizes the formation of interfering chromophores, was used to detect lipid peroxidation. Lipid peroxidation precedes hemolysis and the antioxidant vitamins C and E, which inhibit lipid peroxidation, also inhibit hemolysis. Consequently lipid peroxidation appears to be the cause of RBC destruction. Lipid peroxidation arises mostly from the oxidation of oxyhemoglobin by copper as it is inhibited in RBCs with carbon monoxyhemoglobin or methemoglobin. A direct interaction of copper with the red cell membrane seems to play only a minor role. Copper effects depend on the presence of free SH groups. Lipid peroxidation is probably initiated by activated forms of oxygen as it is increased by an inhibitor of catalase and reduced by hydroxyl radical scavengers. With higher copper concentrations hemolysis is greater: its mechanism appears different as lipid peroxidation is smaller but hemoglobin alterations, namely precipitation, are more pronounced.

Topics: Ascorbic Acid; Copper; Erythrocytes; Ethylmaleimide; Hemolysis; Humans; Lipid Peroxidation; Methemoglobin; Vitamin E

Weanling male Sprague Dawley rats were fed a vitamin E and C-free basal diet with or without supplementation of 100 IU vitamin E per kg diet. After 20 weeks, the vitamin E-deficient rats were divided into four groups, six in each group, and received supplemental ascorbic acid and/or vitamin E by tube feeding daily for 7 days: Group I, 30 mg ascorbic acid/100 g body wt.; Group II, 0.03 mg RRR-alpha-tocopheryl acetate/100 g body wt.; Group III, 30 mg ascorbic acid and 0.03 mg RRR-alpha-tocopheryl acetate/100 g body wt.; and Group IV, placebo. The six control rats (Group V) received placebo. The rats were sacrificed, blood and liver samples were collected for biochemical determinations. Vitamin E deficiency significantly increased erythrocyte (RBC) spontaneous hemolysis, liver thiobarbituric acid (TBA) value, activities of glutamateoxaloacetate transaminase (GOT), pyruvate kinase (PK), and creatine phosphokinase (CPK) in plasma, and significantly lowered plasma vitamin E levels and glutathione peroxidase (GPX) activities. Tube-feeding ascorbic acid for 7 days produced partial reversal effect on liver TBA values, activities of plasma PK, GOT, CPK, and plasma vitamin E levels but not on RBC hemolysis and plasma GPX activity. Tube feeding both ascorbic acid and vitamin E showed similar partial reversal effect as feeding vitamin E alone on all the parameters stated above. The results suggest that ascorbic acid may spare the metabolism of vitamin E and partially reverse the changes in some of the biochemical parameters characteristic of vitamin E deficiency.

Topics: alpha-Tocopherol; Animals; Ascorbic Acid; Aspartate Aminotransferases; Creatine Kinase; Drug Therapy, Combination; Glutathione Peroxidase; Hemolysis; Lipid Peroxides; Liver; Male; Pyruvate Kinase; Rats; Rats, Inbred Strains; Thiobarbiturates; Tocopherols; Vitamin E; Vitamin E Deficiency

Hematoheme displays a potent cytolytic activity toward erythrocytes either in the presence of hydrogen peroxide and a halide ion (system I) or in the presence of oxygen and a reducing agent (system II). In system I it resembles the cytotoxic activity of various peroxidases, whereas in system II it resembles the destructive activity of bleomycin and a variety of metal complexes. Both types of reactions presumably involve the generation of active oxygen species, which are responsible for the damaging effects. In a first attempt to compare the chemical mechanisms of the two types of reactions we used various traps and scavengers of active oxygen species. Tryptophan as well as tyrosine and uric acid were found to be potent inhibitors of the hematoheme-H2O2-halide reaction but do not significantly inhibit the hematoheme-O2-ascorbate reaction. Pyridine, on the other hand, inhibits the oxygen-mediated reaction, but does not affect the peroxide-halide-mediated activity. The cytolytic activity of photoactivated hematoporphyrin, which involves the generation of singlet oxygen, is activated by pyridine and is strongly inhibited by diphenylisobenzofuran. The latter compound is a weak activator of both hematoheme reactions. We conclude that the two hematoheme reactions proceed by two different mechanisms and probably generate different toxic intermediates. The results further suggest that the toxic intermediate generated by photoactivated hematoporphyrin (singlet oxygen) does not play a dominant role in either of the two hematoheme reactions.

Topics: Animals; Ascorbic Acid; Cysteine; Hematoporphyrins; Hemolysis; Hydrogen Peroxide; Iodides; Oxygen; Rabbits; Tryptophan; Uric Acid

Topics: Ascorbic Acid; Carbon Monoxide; Erythrocytes; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Oxidation-Reduction; Peroxides; Plasma; Pyrimidinones; tert-Butylhydroperoxide

Urate, 3-ribosylurate, ascorbate, glutathione and plasma protected bovine, porcine and human erythrocytes from hemolysis caused by t-butyl hydroperoxide (t-BHP). Urate partially protected porcine erythrocytes from hemolysis by t-BHP when it was added 15 min after the addition of the t-BHP, but it did not protect when added 30 min after the t-BHP. Glutathione and ascorbate protected oxyhemoglobin from oxidation to methemoglobin by t-BHP; 3-ribosylurate gave only slight protection. Urate stimulated the formation of methemoglobin from oxyhemoglobin during treatment with t-BHP.

Topics: Animals; Antioxidants; Ascorbic Acid; Cattle; Erythrocytes; Glutathione; Hemolysis; Humans; Kinetics; Male; Methemoglobin; Peroxides; Ribonucleosides; Species Specificity; Swine; tert-Butylhydroperoxide; Uric Acid

The ESR spectrum, attributed to the ascorbic acid (ascorbyl) radical and obtained by exposing freeze dried material to air, can not be used as proof for the occurrence of in vivo free radical reactions. Depending on the method of freeze drying, the content of blood or hemolyzed blood is the dominant factor in creating higher than normal ESR signals in brain or related tissue. These findings explain why the signal, though larger in many human brain tumours than in their surroundings, is not indicative of malignancy. No differences are seen between oedematous and normal tissue. The ascorbyl radical is definitely not stable in aqueous solution, which indicates that fresh tissue sections can also not be used to study in vivo radicals by ESR.

Topics: Animals; Ascorbic Acid; Brain; Brain Neoplasms; Electron Spin Resonance Spectroscopy; Freeze Drying; Hemolysis; Humans; Oxygen; Rats; Rats, Inbred Strains

Topics: Ascorbic Acid; Cardiomyopathies; Erythrocytes; Glutathione Peroxidase; Hair; Hemoglobins; Hemolysis; Humans; Oxidation-Reduction; Selenium; Superoxides

The effect of antioxidants alpha-tocopherol and ionol on membranes of human red cells and bilayer lipid membrane (BLM) from azolektin has been studied. Ionol at concentration 4-10 mM induces the hemolysis of erythrocytes, the cells form changes are observed at concentration 2 mM alpha-tocopherol doesn't show the hemolytic properties at concentration 23 mM. The ionol concentration 1 mM doesn't change the form of the cells, but influence the passive electric parameters: the capacity (Cs) of erythrocytic membrane increases and the intracellular conductance (chi i) decreases. Tocopherol (3 mM) induces the decrease both Cs and chi i. The fast increase of membrane conductance is almost immediately registered on one side of BLM at addition of ionol (0,2-0,4 g/ml). Phosphatidylionol synthesized from ionol and contining the acyl chains C15H31 and C17H35 doesn't influence the electrical properties of BLM.

Topics: Antioxidants; Ascorbic Acid; Butylated Hydroxytoluene; Electric Conductivity; Erythrocyte Membrane; Hemolysis; Humans; Iron; Lipid Bilayers; Membrane Fluidity; Membranes, Artificial; Osmotic Pressure; Phosphatidylcholines; Phospholipids; Vitamin E

The hemolytic effect of saturated fatty acids increased rapidly, when the number of carbon atoms in the chain exceeded 12. At low fatty acid concentrations (less than 60 microM) the hemolytic effect decreased with increasing number of double bonds in the carbon chain (cis-form fatty acids). A more complex pattern was observed at higher fatty acid concentrations. Trans-unsaturated fatty acids were more hemolytic than cis-analogs. Ceruloplasmin, a serum protein with no fatty acid binding capacity, reduced the hemolytic effect of fatty acids; possibly by interacting with the cell membrane. Reducing compounds (thiols, vitamin C) also protected against fatty acid induced hemolysis.

Topics: Animals; Ascorbic Acid; Cattle; Ceruloplasmin; Fatty Acids, Nonesterified; Hemolysis; Humans; Kinetics; Male; Rats; Rats, Inbred Strains; Serum Albumin; Serum Albumin, Bovine; Species Specificity; Sulfhydryl Compounds

As reported previously, the lysate of liquid paraffin-induced guinea pig peritoneal macrophages contains a hemolytic factor which is composed of two components: the soluble (S) and membrane-bound (M) components. To investigate the mechanism whereby the factor hemolysis sheep erythrocytes, an attempt was made to identify the S and M components. The fractionation of the cytosol of macrophages by DEAE-cellulose chromatography and the failure of the lysate from L-ascorbate-depleted macrophages to lyse erythrocytes demonstrated that the S component was L-ascorbate. In addition, L-ascorbate was found to be replaced by NADPH, a substrate of the membrane-bound NADPH oxidase, showing that L-ascorbate acts as a donor of active oxygen. When L-ascorbate was combined with the phospholipids isolated from the membrane fraction by extraction with chloroform-methanol and thin layer chromatography, it became able to lyse erythrocytes. The results so far obtained indicate that the hemolysis by the macrophage lysate is dependent on the formation of peroxidized phospholipids in the membrane fraction with certain active oxygen species produced either from L-ascorbate or by the NADPH oxidase.

Topics: Animals; Ascorbic Acid; Cell Membrane; Chromatography, Thin Layer; Cytosol; Erythrocytes; Guinea Pigs; Hemolysis; Macrophages; Membrane Lipids; NADH, NADPH Oxidoreductases; NADP; NADPH Oxidases; Paraffin; Phospholipids; Sheep

The present study investigates the effect of ascorbate on red cell lipid peroxidation. At a concentration between 0.2 mmol - 20 mmol/1 ascorbic acid reduces hydrogen peroxide-induced red blood cell lipid peroxidation resulting in a marked decrease in ethane and pentane production as well as in haemolysis. Ascorbic acid also shows an antioxidant effect on chelated iron-catalyzed hydrogen peroxide-induced peroxidation of erythrocyte membranes. At a concentration of 10 mmol/1 ascorbic acid totally inhibits oxidative break-down of polyunsaturated fatty acids by radicals originating from hydrogen peroxide. Our results indicate that ascorbate at the chosen concentration has an antioxidant effect on red blood cell lipid peroxidation.

Topics: Ascorbic Acid; Erythrocyte Membrane; Ethane; Fatty Acids; Hemolysis; Humans; Hydrogen Peroxide; In Vitro Techniques; Lipid Peroxides; Oxidation-Reduction; Pentanes

Lysis of human erythrocytes induced by amphotericin B was retarded when the oxygen tension of the incubation mixture was reduced or when the antioxidant catalase was added; lysis was accelerated when cells were preincubated with the prooxidant ascorbate. In the atmosphere of reduced oxygen tension, the erythrocytes containing carboxyhemoglobin lysed at a slower rate than did the cells containing oxyhemoglobin. Consistent with a role for oxidative damage in lysis, the mixture of erythrocytes and amphotericin B showed an increase in malonyldialdehyde, the product of peroxidation, which paralleled the progression of hemolysis. In contrast, the permeabilizing effect of amphotericin B, measured as a decrease in intracellular K+, was not affected by changes in oxygen tension, catalase, or ascorbate treatment. These results imply that oxidant damage is involved in the lytic, but not in the permeabilizing, action of amphotericin B.

Topics: Amphotericin B; Ascorbic Acid; Carboxyhemoglobin; Catalase; Cell Membrane Permeability; Erythrocytes; Hemolysis; Humans; In Vitro Techniques; Light; Malondialdehyde; Oxidation-Reduction; Potassium; Time Factors

Favism is an acute hemolysis occurring in glucose-6-phosphate dehydrogenase (G6PD)-deficient (Mediterranean variant) individuals after intake of fava beans. Divicine (D), 2,6-diamino-4,5-dihydroxypyrimidine, is present in high amounts in the beans, and is suspected to play a role in hemolysis. Its mechanism of action was studied in a cell-free system and in G6PD (Mediterranean variant)-deficient red cells (RBC). Upon hydrolysis of the inactive beta-glucoside vicine, reduced divicine is formed. Oxygen acts as a one- or two-electron acceptor; superoxide anion and hydrogen peroxide are formed, respectively, together with the semiquinoid free-radical form of D. This free radical gives an electron spin resonance (ESR) signal, which is similar to that of the alloxan free radical. Added reduced glutathione (GSH) is rapidly oxidized with a stoichiometry of one to one, and the ESR signal is abolished. Additional GSH is oxidized by hydrogen peroxide and by a slow redox cycle which continuously regenerates oxidized D. The fast-direct and the slow-indirect oxidation result in nonstoichiometric oxidation of GSH. D added to G6PD-deficient RBC rapidly oxidizes GSH with an end point kinetics and a stoichiometry of one to one. Hydrogen peroxide and superoxide anion are scavenged in the RBC and no redox cycling is taking place. No GSH is regenerated even after long incubation periods. After the primary event, i.e., oxidation of GSH and--SH groups, a number of metabolic, rheologic, and membrane modifications, together with increased erythrophagocytosis take place in G6PD-deficient, D-treated RBC only.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Ascorbic Acid; Cell-Free System; Cytochrome c Group; Erythrocytes; Favism; Glucosephosphate Dehydrogenase Deficiency; Glutathione; Hemolysis; Humans; Hydrogen Peroxide; In Vitro Techniques; Kinetics; Male; Oxidation-Reduction; Pyrimidinones; Superoxides

Complete removal of leucocytes and platelets from erythrocytes and the development of a sensitized procedure for the assay of G6PD activity allowed the biochemical mechanisms of the Mediterranean variety of G6PD deficiency to be re-evaluated. Activity in the young erythrocytes from 9 G6PD-deficient subjects averaged 0.1% of the levels observed in the corresponding erythrocyte fraction from normal individuals: moreover, the decline of activity during aging of the G6PD-deficient erythrocytes was comparable with that observed for the normal enzyme. Mutant G6PD purified from granulocytes of a G6PD-deficient subject and entrapped within the corresponding erythrocytes was remarkably stable. Exposure of native erythrocytes to an oxidative stress (divicine plus ascorbate) resulted in a decrease of G6PD activity that was significantly more rapid and extensive in control than in G6PD-deficient cells. These results seem to exclude enhanced intracellular breakdown of the mutant protein within the circulating erythrocytes.

Topics: Ascorbic Acid; Erythrocyte Aging; Erythrocytes; Glucosephosphate Dehydrogenase; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Male; Mediterranean Islands; Mutation; Pyrimidinones

A spectrophotometric method for the determination of serum uric acid based on the oxidation of 2,2'-azino-di(3-ethylbenzthiazoline-6-sulphonate) by use of uricase and peroxidase has already been reported. The method is very precise (CV less than 4.7%). The standard curve is linear up to 4640 mumol/L. Comparison with other enzymatic methods gave good correlation. The method gave results 9% lower than the phosphotungstate method. The effects of bilirubin, haemoglobin, glucose, ascorbic acid, anticoagulants and purine compounds were studied. The reference values for this method are 140.8-407.8 mumol/L for female subjects and 145.6-514.7 mumol/L for male subjects.

Topics: Anticoagulants; Ascorbic Acid; Benzothiazoles; Bilirubin; Blood Glucose; Hemolysis; Humans; Indicators and Reagents; Purines; Reference Values; Sulfonic Acids; Uric Acid

Red blood cells of favism patients with acute hemolytic crisis have markedly more superoxide dismutase (superoxide:superoxide oxidoreductase, EC 1.15.1.1) and less glutathione peroxidase (glutathione:hydrogenperoxide oxidoreductase, EC 1.11.1.9) than either normal controls, glucose-6-phosphate dehydrogenase-deficient subjects or favism patients outside hemolytic crisis. This altered value of the two enzyme activities is not due to increased reticulocyte content of blood. The electrophoretic triplet pattern of superoxide dismutase is also changed, with significant increase of the most positively charged band. Similar modifications of the two enzyme activities are observed after treatment of normal red blood cells with high concentrations of divicine and ascorbate, which are redox compounds that are contained in fava seeds. This treatment produces no hemolysis, but leads to hemolysis if the treated cells are resuspended in the homologous plasma. These results suggest a possible role of active oxygen species in the development of favism.

Topics: Ascorbic Acid; Electrophoresis, Polyacrylamide Gel; Erythrocyte Aging; Erythrocytes; Favism; Glutathione Peroxidase; Hemolysis; Humans; Oxidation-Reduction; Pyrimidinones; Reticulocytes; Superoxide Dismutase

Copper ion induced lysis of rat erythrocytes was markedly stimulated by low concentrations of ascorbate and dehydroascorbate. Ascorbate oxidase, superoxide dismutase, catalase or scavengers of hydroxyl radicals protected erythrocytes against copper-ascorbate stimulated lysis. It is proposed that superoxide radicals and hydrogen peroxide cooperate in producing hydroxyl radicals, which are directly involved in hemolysis. The serum proteins, ceruloplasmin, albumin and apotransferrin, also reduced the hemolytic action of copper-ascorbate, the order of effectiveness being; ceruloplasmin greater than albumin greater than apotransferrin.

Topics: Animals; Apoproteins; Ascorbic Acid; Ceruloplasmin; Copper; Dehydroascorbic Acid; Hemolysis; In Vitro Techniques; Male; Oxidation-Reduction; Rats; Rats, Inbred Strains; Serum Albumin; Transferrin

The addition of sodium nitrite to washed bovine erythrocytes incubated in buffered saline resulted in the formation of methemoglobin with a decrease in the concentration of 3-N-ribosyluric acid. The oxyhemoglobin in hemolysates prepared from bovine red cells which contained high concentrations of 3-ribosyluric acid was oxidized to methemoglobin more slowly than oxyhemoglobin from cells with low levels of 3-ribosyluric acid. Oxyhemoglobin from hemolysates that were dialyzed was oxidized more rapidly than oxyhemoglobin which was not dialyzed. 3-Ribosyluric acid, glutathione, uric acid, and ascorbic acid prevented the oxidation of oxyhemoglobin by nitrite. Uric acid protected oxyhemoglobin at the lowest concentration, followed closely by ascorbic acid, 3-ribosyluric acid, and glutathione. Hydrogen peroxide enhanced the oxidation produced by nitrite; this effect was also prevented by the four antioxidants used.

Topics: Animals; Ascorbic Acid; Cattle; Erythrocytes; Glutathione; Hemolysis; Hydrogen Peroxide; In Vitro Techniques; Methemoglobin; Nitrites; Oxidation-Reduction; Oxyhemoglobins; Ribonucleosides; Sodium Nitrite; Uric Acid

Human caeruloplasmin (ferroxidase), bovine serum albumin and ascorbate protected washed rat erythrocytes against iron ion stimulated haemolysis, while superoxide dismutase, catalase and other scavengers of "activated oxygen" species had little or no effect. Caeruloplasmin retained its protective action when its oxidase activity was completely inhibited by azide, and when its copper ions had been removed. The effect of caeruloplasmin, apocaeruloplasmin and albumin could not be attributed to a binding of iron ions to protein molecules.

Topics: Animals; Apoproteins; Ascorbic Acid; Ceruloplasmin; Erythrocytes; Ferrous Compounds; Hemolysis; Iron; Male; Rats; Serum Albumin; Superoxide Dismutase

Male rats fed on pellet diet to an average weight of 105 g were placed on a vitamin E deficient diet containing 20% coconut oil for a period of 12 weeks at two dietary protein levels, 20% and 10% casein. Rats on 20% casein diet showed a definite weight loss but not so at the 10% casein level. A marked increase in the liver in vitro lipid peroxidation was observed at both protein levels. Feeding of retinyl palmitate at 100,000 IU/100 g body weight for 4 consecutive days inhibited the liver, brain and kidney in vitro peroxidation; megadoses of ascorbic acid produced less inhibition of the liver peroxidation, but the same degree of inhibition for brain and kidney peroxidation as in vitamin A loaded rats. Both dietary palmitate or ascorbic acid. Acetylcholine esterase and ATPase, two of the membrane enzymes of erythrocytes, were depressed in all the groups. The glutathione content of erythrocytes was increased in rats given ascorbic acid. In all the groups the higher dietary protein levels produced greater loss of body and tissue weights. It is concluded vitamin E deficient diet supplemented with dietary coconut oil (saturated fat) induces increased in vitro lipid peroxidation and oxidative lysis of erythrocytes and that megadoses of vitamin A or C suppress the in vitro lipid peroxidation but enhance the lysis.

Topics: Animals; Ascorbic Acid; Brain; Caseins; Cocos; Dietary Fats; Dose-Response Relationship, Drug; Hemolysis; Kidney; Lipid Peroxides; Liver; Male; Organ Size; Rats; Vitamin A; Vitamin E Deficiency

Topics: Ascorbic Acid; Erythrocytes; Formates; Hemolysis; Humans; In Vitro Techniques; Methemoglobin; Primaquine

Topics: Animals; Ascorbic Acid; Azides; Dithiothreitol; Erythrocytes; Goats; Hemolysis; Polysorbates; Rats; Vitamin E; Vitamin E Deficiency

It is proposed that the loss of ability by humans to synthesize ascorbic aicd may have markedly enhanced the survival opportunities of early man living in a malarial infested environment. This hypothesis is based on biomedical evidence which indicates that glucose-6-phosphate dehydrogenase (G-6-PD) deficient individuals display enhanced sensitivity to ascorbic acid induced hemolysis which has been fatal at sufficiently high doses and that the G-6-PD deficient trait has been selected for in malarial environments.

Topics: Ascorbic Acid; Biological Evolution; Genetics, Population; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Malaria; Species Specificity

Topics: Animals; Ascorbic Acid; Catalase; Erythrocytes; Hemolysis; In Vitro Techniques; Lipid Peroxides; Male; Oxygen; Rabbits; Sodium Chloride; Superoxide Dismutase; Vitamin E; Zinc

Topics: Ascorbic Acid; Erythrocytes; Glucosephosphate Dehydrogenase; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Infant, Newborn; Jaundice, Neonatal

Rats were maintained on a vitamin E free diet containing 20% safflower oil for a period of 12 weeks at two dietary protein levels, 20% and 10% casein. Enhanced in vitro tissue lipid peroxidation and lysis of erythrocytes were noticed at both the protein levels. A reduction in body mass and tissue weights were observed in both the protein groups but more so at 20% protein level. Feeding of retinyl palmitate (100 000 IU/100 g body weight) for 4 consecutive days to -E rats inhibited liver and kidney in vitro lipid peroxidation. Ascorbic acid (150 mg/100 g body weight) given orally for 5 days to -E rats inhibited liver brain and kidney in vitro peroxidation. Lysis of erythrocytes from -E rats was further increased by dosing with both the vitamins "A" and "C", the latter being more effective. The stromal enzymes acetyl choline esterase and ATPase were lowered, following the hemolysis profile of the erythrocytes from the different groups. Glutathione content of erythrocytes were unaffected except in -E +C group. In all groups the higher protein level (20%) produced greater lysis as compared to 10% level. It is concluded that 20% protein is more injurious in vitamin E deficiency simultaneously made hypervitaminosis A or C.

Topics: Animals; Ascorbic Acid; Dietary Fats; Dietary Proteins; Diterpenes; Hemolysis; Lipid Peroxides; Male; Oxidation-Reduction; Rats; Retinyl Esters; Vitamin A; Vitamin E Deficiency

Topics: Anemia, Sickle Cell; Animals; Ascorbic Acid; Copper; Hemolysis; Humans; Rats; Risk; Sheep; Sickle Cell Trait

Weanling rats were maintained on purified diets with graded levels of vitamin E at 50, 60, 100, 200, or 500 IU/kg diet; each group was further divided into two subgroups, each with six rats and received vitamin C at 0 or 1.5 g/kg diet. After 1 or 2 months, the determination of antioxidant status showed that the high supplementation of vitamin C at the marginally adequate vitamin E level significantly increased in vitro erythrocyte hemolysis and liver lipid peroxidation, and significantly lowered erythrocyte level of reduced glutathione and plasma level of vitamin E; thus lowering the overall antioxidant potential of the animals. A small increase in vitamin E level counteracted the hemolytic and peroxidative effect of the high supplementation of vitamin C. A greater increase in vitamin E level counteracted the effect of high supplementation of vitamin C in decreasing glutathione level and plasma vitamin E level. These results indicate that the adverse effect of the high supplementation of vitamin C on tissue antioxidant potential may be overcome by increasing the supplementation level of vitamin E, and suggest that vitamin E requirement may be increased with increased vitamin C supplementation.

Topics: Animals; Antioxidants; Ascorbic Acid; Dose-Response Relationship, Drug; Hemolysis; Lipid Peroxides; Male; Nutritional Requirements; Rats; Time Factors; Vitamin E

Hemolytic sensitivity of various density (and age) fractions of bovine erythrocytes to oxidant stress induced by H2O2, ascorbate and gamma radiation, as well as to prolonged incubation (autohemolysis) and heat treatment was compared. The sensitivity of older red cells was increased for higher but not for lower rates of "oxidative load" nor to autohemolysis and hyperthermic hemolysis. Changes in the sensitivity of senescent erythrocytes to redox agents are mediated, at least partly, by factors other than glutathione concentration which was fairly constant during the intravascular red cell aging.

Topics: Animals; Ascorbic Acid; Cattle; Cobalt Radioisotopes; Erythrocyte Aging; Erythrocytes; Hemolysis; Hot Temperature; Hydrogen Peroxide; In Vitro Techniques

The preventive effects of L-ascorbic acid (V.C) and pantothenic acid (PaA) against photosensitized hemolysis in the presence of pheophorbide a or hematoporphyrin were investigated in vitro. The development of hematoporphyrin-induced photosensitivity and the preventive effects of V.C and PaA were also investigated in male mice. V.C and PaA exerted preventive effects against photosensitized hemolysis in the presence of pheophorbide a or hematoporphyrin. In such cases, V.C and PaA had not only an additive effect but also a synergistic one. The time required for the photosensitized acute shock-like death (i.e., survival time), was significantly extended by simultaneous administration of V.C and PaA. This supports the findings of in vitro experiments.

Topics: Animals; Ascorbic Acid; Chlorophyll; Drug Synergism; Hematoporphyrins; Hemolysis; Light; Male; Mice; Pantothenic Acid; Photosensitivity Disorders; Rats

Topics: Adult; Anemia, Hemolytic; Ascorbic Acid; Female; Hemolysis; Humans; Hypochlorous Acid; Japan; Kidney Failure, Chronic; Male; Renal Dialysis; Water Supply

Two groups of weanling male Hartley guinea pigs maintained on vitamin E deficient diet were supplemented with 0.4 I.U./100 g body weight/day of vitamin E and either 2 (Group A) or 10 (Group B) mg/100 g body weight/day of vitamin C for 5 weeks. As compared to Group A, the degree of erythrocyte hemolysis and liver TBAR level of Group B were significantly increased while plasma vitamin E and erythrocyte GSH levels were significantly decreased. In another experiment, two groups of guinea pigs were given 0.8 I.U./100 g body weight/day of vitamin E and 2 (Group C) or 30 mg/100 g body weight/day (Group D) of vitamin C. Levels of plasma vitamin E and erythrocyte GSH of Group D were significantly lower than those of Group C: however, erythrocyte hemolysis and liver TBAR were not affected by the level of vitamin C supplementation. The results suggest that the high levels of vitamin C supplementation lowered tissue antioxidant potential of animal when vitamin E was marginally adequate, and the hemolytic and peroxidizing effect of high level of vitamin C may be counteracted by increasing the level of vitamin E.

Topics: Animals; Antioxidants; Ascorbic Acid; Erythrocytes; Glutathione; Guinea Pigs; Hemolysis; Liver; Male; Thiobarbiturates; Vitamin E; Vitamin E Deficiency

Chloramines, compounds made up of chlorine and ammonia, when present in tap water used for dialysis cause methemoglobinemia and hemolysis. Ascorbic acid addition has been reported to effectively neutralize chloramines in vitro and in patients dialyzed with the single batch dialysis delivery system. We extended these observations to patients dialyzed with the proportioning dialysis delivery system where exposure time of ascorbic acid to chloramines is shorter. This may be important since we found that the half time of the reaction between ascorbic acid and chloramines is 4 minutes. Red cell oxidant sensitivity in 15 patients was assessed by incubating red cells with ascorbate-cyanide and measuring methemoglobin which averaged 2.17 +/- 0.42 g/100 ml (SEM) before dialysis and 2.87 +/- 0.52 g/100 ml after dialysis (NS). Reduced glutathione (GSH) levels were also measured as an index of red cell oxidant damage. GSH decreased from a mean of 7.40 +/- 0.59 micromoles/g Hb before dialysis to 6.98 +/- 0.52 micronmoles/g Hb after dialysis (P less than 0.01). In 2 patients there was no change in 51Cr red cell survival when dialyzed on either the proportioning system or other chloramine free systems. We conclude that addition of ascorbic acid to neutralize chloramines in tap water is also effective when using the proportioning dialysis delivery system.

Topics: Adult; Aged; Ascorbic Acid; Chloramines; Erythrocyte Aging; Glutathione; Hemolysis; Humans; Kidney Failure, Chronic; Methemoglobin; Methemoglobinemia; Middle Aged; Renal Dialysis

The effect of dietary vitamin C on vitamin E status and function was studied with weanling male Hartley guinea pigs. Thirty guinea pigs fed a basal diet free of vitamins E and C were divided into two groups; only one group received 2 mg/100 g body weight/day of vitamin C. Each group was further divided into three subgroups and received 0, 0.2, and 0.4 IU/100 g body weight/day of vitamin E. After 5 weeks the animals were killed. Degree of erythrocyte hemolysis induced by dialuric acid, level of plasma vitamin E and extent of liver lipid peroxidation were determined. Liver mitochondria were isolated and in vitro oxygen uptake was measured. The rate of swelling of mitochondria in two media was also determined. The results showed that the adequate dietary vitamin C supplementation did not affect the extent of erythrocyte hemolysis induced by dialuric acid, but increased the plasma vitamin E level and lowered liver lipid peroxidation. It also maintained the normal respiration and swelling rates of liver mitochondria when vitamin E was inadequate.

Topics: Animals; Ascorbic Acid; Dose-Response Relationship, Drug; Guinea Pigs; Hemolysis; Lipid Metabolism; Male; Mitochondria, Liver; Mitochondrial Swelling; Oxygen Consumption; Peroxides; Vitamin E

Topics: Administration, Oral; Animals; Ascorbic Acid; Brain; Hemolysis; Injections, Intraperitoneal; Kidney; Linoleic Acids; Lipid Metabolism; Liver; Rats; Spleen; Vitamin A

We describe the effects of uric acid, hemolysis, drugs, ascorbic acid, lipemia, and bilirubin on the enzymic measurement of cholesterol in serum by use of reagent kits from Abbott, Beckman, Boehringer Mannheim, Calbiochem, and Worthington. In all of these, the chromogen formed from the reaction of hydrogen peroxide with phenol and 4-aminoantipyrene is measured. The absorbance was measured at 500 nm vs. a serum blank for each kit--except Abbott's with which the recorded absorbances were the differences between readings at 500 and 600 nm. With all reagents kits, there was no interference from uric acid up to 200 mg/liter, hemoglobin up to 1.0 g/liter, or drugs (clofibrate, phenobarbital, Ketochol, Ovral-28), but negative interferences from ascorbic acid. Except for the Abbott kit, the cholesterol values obtained for lipemic samples were lower than found with the comparison method [Abell et al., Stand. Methods Clin. Chem. 2, 26 (1958)]. With Abbott's reagents, for most lipemic samples, the values were the same. Bilirubin at concentrations of 200 mg/liter significantly decreased the cholesterol values with Beckman, Calbiochem, and Worthington reagent kits. With Boehringer Mannheim reagent a small negative interference was observed and with Abbott reagent a small positive interference was observed when the bilirubin concentrations were 200 mg/liter.

Topics: Ascorbic Acid; Bilirubin; Cholesterol; Evaluation Studies as Topic; Hemolysis; Indicators and Reagents; Lipids; Spectrophotometry; Surface-Active Agents; Uric Acid

Topics: Ascorbic Acid; Blood Specimen Collection; Erythrocytes; Folic Acid; Freezing; Hemolysis; Humans; Methods; Radioligand Assay

Topics: Ascorbic Acid; Burns; Dose-Response Relationship, Drug; Erythrocytes; Female; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Male; Middle Aged; Uric Acid

Topics: Ascorbic Acid; Blood Glucose; Carbon Dioxide; Cyanides; Erythrocytes; Glucosephosphate Dehydrogenase Deficiency; Glutathione; Hemoglobins; Hemolysis; Humans; In Vitro Techniques; NADP; Oxygen

Topics: Aged; Ascorbic Acid; Black People; Burns; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Male

Topics: Animals; Ascorbic Acid; Dose-Response Relationship, Drug; Drug Compounding; Glycerol; Hemolysis; Injections, Intravenous; Mice; Rabbits; Solutions

Topics: Anaerobiosis; Animals; Ascorbic Acid; Cells, Cultured; Culture Media; Cystine; Erythrocytes; Glutathione; Hemolysis; Horseradish Peroxidase; Hot Temperature; Macrophages; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Molecular Weight; Oxygen; Sulfhydryl Reagents; Vitamin E

Topics: Anemia, Pernicious; Ascorbic Acid; Biological Assay; Densitometry; Erythrocytes; Folic Acid; Folic Acid Deficiency; Hematocrit; Hemolysis; Humans; Lacticaseibacillus casei; Methods

The hemolytic activity of fibrous asbestos varieties and of fibrous or granular silica dust can be markedly reduced by adsorption of polymers. Polyanions exert a specific action on asbestos, particularly chrysotile, whereas silica is inactivated by nonionic polymers. A high degree of reduction of the lytic action by comparatively small amounts of the antagonistic polymers can be demonstrated after short exposure to concentrations of 0.1-0.4 mg/ml of appropriate polymers. Inactivation is based on stable adsorption. Repeated washings of inactivated mineral sediments or exposure to elevated temperatures (80-120 degrees C) produced no essential loss of the reduction of lytic potency. In one example, inactivation of chrysotile by sodium alginate, depolymerization by ascorbic acid was also ineffective.

Topics: Adsorption; Alginates; Asbestos; Ascorbic Acid; Carboxymethylcellulose Sodium; Chondroitin Sulfates; Hemolysis; Hot Temperature; Povidone; Silicon Dioxide; Surface Properties; Time Factors

Topics: Absorption; Ascorbic Acid; Bilirubin; Color; Digoxin; Hemolysis; Hydrogen Peroxide; Iodine Radioisotopes; Quaternary Ammonium Compounds; Radioimmunoassay; Scintillation Counting; Triiodothyronine; Tritium

Topics: Ascorbic Acid; Azo Compounds; Bilirubin; Chemistry, Clinical; Diazonium Compounds; Epinephrine; Hemoglobins; Hemolysis; Histidine; Humans; Hyperlipidemias; Levodopa; Methyldopa; Norepinephrine; Rifampin; Spectrophotometry, Ultraviolet; Statistics as Topic; Tyrosine

Topics: Anemia, Hemolytic; Ascorbic Acid; Cell Survival; Charcoal; Chloramines; Chromium Radioisotopes; Erythrocytes; Filtration; Hemolysis; Hexosephosphates; Humans; Methemoglobin; Osmosis; Renal Dialysis; Uremia; Water Pollution, Chemical; Water Supply; Water-Electrolyte Balance

Topics: Ascorbic Acid; Blood; Carbon Radioisotopes; Enzyme Activation; Erythrocytes; Flavin-Adenine Dinucleotide; Flavins; Glucosephosphate Dehydrogenase Deficiency; Glutathione Reductase; Hemolysis; Humans; Liver Cirrhosis; Methylene Blue; Riboflavin; Umbilical Cord; Uremia

Topics: Ascorbic Acid; Autoanalysis; Bilirubin; Evaluation Studies as Topic; Hemolysis; Humans; Indicators and Reagents; Methods; Microchemistry; Oxygen Consumption; Polarography; Spectrophotometry; Time Factors; Urate Oxidase; Uric Acid

Topics: Ascorbic Acid; Autoanalysis; Blood Proteins; Child; Child, Preschool; Dialysis; Dinitrophenols; Evaluation Studies as Topic; Fasting; Hemolysis; Humans; Hydrazines; Hydrogen-Ion Concentration; Indicators and Reagents; Indophenol; Mass Screening; Methods; Microchemistry; Plasma; Spectrophotometry; Time Factors

Topics: Amitrole; Animals; Ascorbic Acid; Catalase; Cell Survival; Chromium Isotopes; Dapsone; Erythrocytes; Galactose; Glucosephosphate Dehydrogenase Deficiency; Glutathione; Hemolysis; Humans; Hydrogen Peroxide; Hydroxylamines; Methemoglobin; Rats

Topics: Adult; Ascorbic Acid; Bilirubin; Blood Glucose; Erythrocytes; Female; Fluorides; Glucose; Glucose Oxidase; Glycosuria; Hematocrit; Hemolysis; Humans; Methods; Middle Aged; Peroxidases; Sodium; Time Factors; Uric Acid

Topics: Ascorbic Acid; Erythropoiesis; Hemolysis; Humans; Porphyrias; Vitamin E

Topics: Ascorbic Acid; Autoanalysis; Buffers; Color; Copper; Dialysis; Hemolysis; Humans; Indicators and Reagents; Iron; Methods; Photometry

Topics: Anemia, Hemolytic; Ascorbic Acid; Carbon Radioisotopes; Cell Survival; Chromium Radioisotopes; Erythrocytes; Glucose; Glucosephosphate Dehydrogenase Deficiency; Heinz Bodies; Hemolysis; Hexosephosphates; Humans; Metabolism, Inborn Errors; Primaquine; Renal Dialysis; Sulfonamides; Uremia; Water

Topics: Ascorbic Acid; Autoanalysis; Colorimetry; Hemoglobins; Hemolysis; Humans; Iron; Iron Chelating Agents; Methods; Pyridines; Sulfonic Acids; Transferrin; Triazines

Topics: Animal Nutritional Physiological Phenomena; Animals; Ascorbic Acid; Cell Membrane; Deficiency Diseases; Drug Synergism; Erythrocytes; Globins; Glucose; Glutathione; Heinz Bodies; Hemoglobins; Hemolysis; Male; Methemoglobin; Oxidation-Reduction; Porphyrins; Rats; Selenium; Vitamin E; Vitamin E Deficiency

Topics: Aniline Compounds; Ascorbic Acid; Azides; Blood Glucose; Catalase; Chlorates; Erythrocytes; Glucosephosphate Dehydrogenase; Glutathione; Heinz Bodies; Hemoglobinometry; Hemolysis; Humans; Methemoglobin; Nitrites; Osmotic Fragility; Peroxidases; Phenylhydrazines; Potassium; Primaquine; Sodium; Vitamin K

Topics: Aldehydes; Antioxidants; Ascorbic Acid; Azides; Catalase; Emulsions; Erythrocytes; Glutathione; Hemolysis; Humans; Hydrogen Peroxide; Hydrogen-Ion Concentration; Lipids; Malonates; Nucleosides; Oxidation-Reduction; Oxygen; Phosphatidylcholines; Serum Albumin, Bovine; Spectrum Analysis; Time Factors

Topics: Adult; Ascorbic Acid; Carbon Dioxide; Carbon Isotopes; Erythrocytes; Erythrocytes, Abnormal; Female; Glucose; Glucosephosphate Dehydrogenase; Glucosephosphate Dehydrogenase Deficiency; Glutathione; Glutathione Reductase; Heinz Bodies; Hemolysis; Hexosephosphates; Humans; Methylene Blue; Peroxidases; Peroxides; Pyruvate Kinase; Sodium Chloride; Urinary Tract Infections

Topics: Ascorbic Acid; Bilirubin; Catalase; Color; Colorimetry; Glucose; Hemolysis; Humans; Mathematics; Methods; Ultraviolet Rays; Urate Oxidase; Uric Acid

Topics: Amino Acids; Animals; Ascorbic Acid; Blood; Blood Glucose; Blood Proteins; Blood Volume Determination; Chromium; Chromium Radioisotopes; Citrates; Dogs; Edetic Acid; Erythrocytes; Glucose; Hematocrit; Hemoglobins; Hemolysis; Heparin; Humans; Hydrogen-Ion Concentration; Ion Exchange; Methods; Oxidation-Reduction; Riboflavin; Rutin

Topics: Adult; Ascorbic Acid; Equipment and Supplies; Erythrocytes; Hemolysis; Humans; Light; Nitrogen; Osmolar Concentration; Oxygen; Porphyrias; Potassium; Radiation Effects

Topics: Ascorbic Acid; Aspirin; Blood Donors; Glucosephosphate Dehydrogenase Deficiency; Hemolysis; Humans; Morocco

Topics: Ascorbic Acid; Blood; Carcinoma; Catalase; Conductometry; Dialysis; Edetic Acid; Erythrocytes; Ethylmaleimide; Hemolysis; Hot Temperature; Humans; Kidney; Liver; Lung Neoplasms; Male; Neoplasms; Prostatic Neoplasms; Stomach Neoplasms; Tissue Extracts; Triazoles

Topics: Adipose Tissue; Animals; Ascorbic Acid; Atmospheric Pressure; Brain; Carbon Isotopes; Hemolysis; Lipid Metabolism; Liver; Male; Oxygen; Peroxides; Rats; Stress, Physiological; Sulfhydryl Compounds; Vitamin E; Vitamin E Deficiency

Topics: Aminocaproates; Ascorbic Acid; Erythrocytes; Gluconates; Hemolysis; Humans; Peptides; Staphylococcus

Topics: Acatalasia; Anemia; Anemia, Hemolytic; Ascorbic Acid; Azides; Catalase; Cyanides; Erythrocyte Aging; Erythrocytes; Ethylmaleimide; Genetics, Medical; Glucose; Glucosephosphate Dehydrogenase Deficiency; Glucosephosphates; Glutathione; Hemoglobins; Hemolysis; Hexosephosphates; Humans; Metabolism, Inborn Errors; Methemoglobin; Peroxides; Pharmacology; Phenylhydrazines; Primaquine; Sulfhemoglobin; Sulfhydryl Compounds

Topics: Anemia; Anemia, Pernicious; Ascorbic Acid; Clinical Enzyme Tests; Dehydroascorbic Acid; Erythrocytes; Hemolysis; Humans; Oxidoreductases; Vitamin B 12

Topics: Adenosine Triphosphate; Anemia; Anemia, Hemolytic; Ascorbic Acid; Blood Chemical Analysis; Blood Glucose; Carbon Dioxide; Erythrocyte Count; Erythrocytes; Geriatrics; Glucosephosphate Dehydrogenase; Glutathione; Hemoglobinometry; Hemolysis; Kidney Diseases; Metabolism; Methemoglobinemia; Nucleosides; Pyelonephritis; Renal Insufficiency; Sulfamethizole; Sulfathiazoles; Sulfonamides; Toxicology; Urinary Catheterization

Topics: Anemia; Anemia, Macrocytic; Ascorbic Acid; Ascorbic Acid Deficiency; Blood Cell Count; Blood Chemical Analysis; Blood Coagulation Tests; Blood Proteins; Bone Marrow Examination; Folic Acid; Gastric Acidity Determination; Geriatrics; Hemoglobins; Hemolysis; Hemorrhagic Disorders; Humans; Scurvy; Urine; Urobilin

Topics: Animals; Ascorbic Acid; Cats; Cattle; Glutathione; Guinea Pigs; Hemolysis; Liver; Metabolism; Mice; Pharmacology; Rats; Sheep

Topics: Abruptio Placentae; Ascorbic Acid; Blood Chemical Analysis; Female; Hemolysis; Humans; Metabolism; Pre-Eclampsia; Pregnancy

Topics: Animals; Ascorbic Acid; Cell Death; Erythrocytes; Guinea Pigs; Hemolysis; Osmosis; Research; Vitamins

Topics: Ascorbic Acid; Hemolysis; Humans; Oxygen; Vitamins

Topics: Ascorbic Acid; Hemolysis; Humans; Temperature

Topics: Ascorbic Acid; Erythrocytes; Hemolysis; Humans; Osmotic Fragility