oxalates and Hemolysis

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

Oxalate is a minor, but important metabolite of ethylene glycol and has been directly linked with acute and subchronic renal toxicity in ethylene glycol poisoning. Numerous studies have characterized the cytotoxicity of oxalate as including plasma membrane damage and organelle injury. Oxalate has two forms in vivo: oxalate ions and calcium oxalate monohydrate (COM) crystals that readily form in the presence of calcium. The present study was designed to compare the cytotoxicity of the oxalate ion and COM crystals in human and rat cells. In rat red blood cells, the oxalate ion did not increase hemolysis, while COM crystals produced hemolysis with a concentration-dependent increase. In human proximal tubule (HPT) cells in culture, COM suspensions, at concentrations >3 mM but with no oxalate ion, caused cytotoxicity as evidenced by the release of lactate dehydrogenase (LDH) into media. Cytotoxicity was not observed in HPT cells treated with oxalate solutions that contained no COM because EDTA prevented its formation. The cytotoxic effects of COM to HPT cells were potentiated by acidosis (pH 6.5), but not by glycolate, the major metabolite of ethylene glycol. The toxicity of COM to HPT cells and to proximal tubule cells from Wistar and F-344 rats, compared using both ethidium homodimer uptake and LDH leakage, increased in human and rat cells in a concentration-dependent manner. Rat cells were more sensitive to COM than HPT cells, but there were no apparent differences between the effects in Wistar cells and F-344 cells. These results demonstrate that COM crystals, and not the oxalate ion, are responsible for the membrane damage and cell death observed in normal human and rat PT cells and suggest that COM accumulation in the kidney is responsible for the renal toxicity associated with ethylene glycol exposure.

Topics: Animals; Calcium Oxalate; Cell Death; Cells, Cultured; Chelating Agents; Crystallization; Dose-Response Relationship, Drug; Edetic Acid; Ethylene Glycol; Hemolysis; Humans; Kidney Tubules, Proximal; Oxalates; Oxalic Acid; Rats; Rats, Inbred F344; Rats, Wistar; Species Specificity

The precise nature of band 3 protein and its involvement in oxalate exchange in the red blood cells (RBCs) of renal failure patients has not been studied in detail. Therefore, here we studied the oxalate exchange and binding by band 3 protein in RBCs of humans with conditions of acute and chronic renal failure (ARF and CRF). The RBCs of ARF and CRF patients exhibited abnormal red cell morphology and an increased resistance to osmotic hemolysis. Further, an increase in the cholesterol content and decrease in the activities of Na(+)-K(+)-, Ca(2+)-, and Mg(2+)-ATPases of membranes were observed in the RBCs of ARF and CRF patients. A decrease in the oxalate flux was observed in the RBCs of ARF and CRF patients. The oxalate-binding activities of the RBC membranes were significantly lower in ARF (20 pmoles/mg protein) and CRF (5.3 pmoles/mg protein) patients as compared to that in the normal subjects (36 pmoles/mg protein). DEAE-cellulose and Sephadex G-200 column chromatography purification profiles revealed a distinctive shift in oxalate-binding activity of band 3 protein of RBCs of ARF and CRF patients as compared to that of the normal subjects. It was also observed from the binding studies with a fluorescent dye, eosin-5-maleimide, which specifically binds to band 3 protein, that the RBCs of ARF and CRF patients exhibited only 53 and 32% of abundance of band 3 protein, respectively, as compared to that in the RBCs of the normal subjects, thus revealing a decrease in the band 3 protein content in ARF and CRF patients. These results for the first time showed a decrease in the oxalate exchange in RBCs of patients with ARF and CRF, which was also concomitant with the low levels of abundance of band 3 protein.

Topics: Acute Kidney Injury; Anion Exchange Protein 1, Erythrocyte; Eosine Yellowish-(YS); Erythrocyte Membrane; Erythrocytes; Female; Fluorescent Dyes; Hemolysis; Humans; Kidney Failure, Chronic; Male; Middle Aged; Oxalates

Topics: Animals; Anticoagulants; Cattle; Citrates; Edetic Acid; Erythrocytes; Hemolysis; Heparin; Humans; Male; Osmotic Fragility; Oxalates; Sodium Citrate

1. Studies were conducted to determine the effects of anticoagulants and storage (4 degrees C) on the PCV of blood samples from Nigerian domestic fowl (DF) and the guinea fowl (GF). 2. Citrate significantly reduced pre-storage PCV of the DF in comparison with the effect of ethylenediamine tetra-acetic acid (EDTA). 3. It further decreased (P < 0.05) the PCV of the blood of DF and GF over 3 d of storage; this was similar to the effect of EDTA on the PCV of the GF blood. 4. Citrate and oxalates induced haemolysis of blood of the DF and the GF in storage faster than EDTA, but overall the haemolysis was more pronounced from red cells of the GF than from those of the DF. 5. The mean fall in the PCV of the DF was significant at 3.0 mg EDTA/ml of blood in contrast to the fall in the PCV of the GF blood.

Topics: Animals; Anticoagulants; Citrates; Edetic Acid; Erythrocytes; Hematocrit; Hemolysis; Nigeria; Oxalates; Poultry; Temperature

The effects of 3 UICC asbestos fibres (A chrysotile, crocidolite, amosite) were observed in vitro on red blood cells (RBC) and alveolar macrophages (AM). THe reactivity of the fibres after leaching with 0.1 N oxalic acid or adsorption of SO2 or benzo-3,4-pyrene (BP) was studied. The haemolytic activity of crocidolite and amosite was very low. A cytotoxic effect on AM occurred when the fibres were present in high concentration (100 microgram/ml), this was characterized by a release of both cytoplasmic (LDH) and lysosomal (beta-galactosidase) enzymes. The leached fibres were more haemolytic than the unleached ones, and more beta-galactosidase (beta-Gal) than lactic dehydrogenase (LDH) was released from the AM. In contrast to the amphiboles, chrysotile fibres were highly haemolytic and induced a selective release of beta-Gal from AM. Leached fibres were less haemolytic and were cytotoxic for AM (both enzymes were released). Their in vitro reactivity was similar to that observed with quartz. The results showed that SO2 changed the reactivity very little. BP sorption on acid-leached chrysotile decreased the LDH release from AM. The difference in the in vitro reactivity related to the chemical state of asbestos fibres might explain the difference in their in vivo reactivity (latency, degree of fibrosis). This point is discussed.

Topics: Animals; Asbestos; Benzo(a)pyrene; Benzopyrenes; beta-Galactosidase; Erythrocytes; Hemolysis; Humans; In Vitro Techniques; L-Lactate Dehydrogenase; Macrophages; Oxalates; Pulmonary Alveoli; Rabbits; Sulfur Dioxide

Topics: Autoanalysis; Drug Stability; Edetic Acid; Erythrocytes; Evaluation Studies as Topic; Fluorides; Glutathione; Hematocrit; Hemolysis; Humans; Hydro-Lyases; Indicators and Reagents; Kinetics; Methods; Oxalates; Oxidation-Reduction; Porphobilinogen Synthase; Saponins; Spectrophotometry; Temperature; Time Factors; Ultrasonics

Topics: Animals; Anticoagulants; Blood Glucose; Cattle; Edetic Acid; Erythrocyte Count; Fluorides; Glycolysis; Hematocrit; Hemoglobinometry; Hemolysis; Oxalates; Potassium; Quaternary Ammonium Compounds; Sodium; Temperature; Time Factors

Topics: Animals; Bacteria; Blood; Cattle; Citrates; Corynebacterium pyogenes; Culture Media; Depression, Chemical; Dose-Response Relationship, Drug; Escherichia coli; Haemophilus; Hemolysis; Methods; Moraxella; Oxalates; Potassium; Sodium; Staphylococcus; Streptococcus

Topics: Animals; Bicarbonates; Carbon Isotopes; Dicarboxylic Acids; Hemolysis; Humans; Iron; Iron Isotopes; Kinetics; Malonates; Oxalates; Rabbits; Reticulocytes; Spectrophotometry; Temperature; Transferrin

Topics: Animals; Edetic Acid; Fluorides; Hemoglobins; Hemolysis; Humans; Insulin; Iodine Isotopes; Methods; Oxalates; Radioimmunoassay; Swine

Topics: Blood Sedimentation; Bone Marrow; Bone Marrow Cells; Centrifugation; Hemolysis; Humans; Leukemia, Lymphoid; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Leukocytes; Leukocytosis; Methods; Oxalates; Quaternary Ammonium Compounds; Staining and Labeling

Topics: Biotransformation; Chromatography, Paper; Dihydroxyphenylalanine; Dopamine; Erythrocytes; Fluorescence; Hemolysis; Heparin; Humans; Male; Melanins; Norepinephrine; Oxalates; Spectrophotometry; Time Factors; Ultraviolet Rays

Topics: Chromatography, Paper; Chronic Disease; Dihydroxyphenylalanine; Dopamine; Erythrocytes; Hemolysis; Heparin; Humans; Male; Melanins; Norepinephrine; Oxalates; Schizophrenia; Spectrophotometry; Ultraviolet Rays

Topics: Animals; Anticoagulants; Blood Preservation; Blood Sedimentation; Blood Viscosity; Citrates; Edetic Acid; Hemolysis; Heparin; Horse Diseases; Horses; Oxalates; Temperature

Topics: ABO Blood-Group System; Blood Transfusion; Citrates; Dialysis; Erythrocytes; Hematocrit; Hemoglobinometry; Hemoglobinuria, Paroxysmal; Hemolysis; Humans; Immune Sera; Isotonic Solutions; Oxalates; Sucrose; Temperature

Topics: Adrenochrome; Adult; Chromatography, Paper; Chronic Disease; Epinephrine; Erythrocytes; Fluorescence; Hemolysis; Heparin; Humans; Indoles; Male; Melanins; Middle Aged; Oxalates; Schizophrenia; Spectrum Analysis

Topics: Adrenochrome; Chromatography, Paper; Epinephrine; Fluorescence; Heme; Hemolysis; Heparin; Humans; In Vitro Techniques; Male; Melanins; Oxalates; Plasma; Porphyrins; Spectrum Analysis

Topics: Anti-Bacterial Agents; Bacteriological Techniques; Blood; Calcium; Complement System Proteins; Enterobacter aerogenes; Escherichia coli; Hemagglutination; Hemagglutination Tests; Hemolysis; Humans; Microbiology; Muramidase; Oxalates; Pharmacology; Plasma; Research