Nitrite-induced Methemoglobin Formation Research Articles

Taurine mitigates nitrite-induced methemoglobin formation and oxidative damage in human erythrocytes.

Nitrite is present as a noxious contaminant in drinking water and causes oxidative damage in various tissues of humans and animals. It is a well-known methemoglobin-forming agent that has been shown to damage blood cells. The protective effect of taurine, a semi-essential sulfur-containing amino acid, was studied on sodium nitrite (NaNO2)-induced oxidative damage in human erythrocytes. Erythrocytes were incubated with NaNO2, in the presence and absence of taurine, and changes in oxidative stress parameters determined. Pretreatment with taurine significantly ameliorated NaNO2-induced oxidative damage to lipids, proteins, and plasma membrane. It also reduced the NaNO2-induced increase in methemoglobin levels and ROS production. Taurine improved the antioxidant capacity of cells, restored the alterations in the activities of various metabolic enzymes, and prevented morphological changes in erythrocytes. Thus, taurine can be potentially used as a protective agent against the damaging effects of nitrite.

Crocin protects human erythrocytes from nitrite-induced methemoglobin formation and oxidative damage.

Sodium nitrite (NaNO2 ) is a common contaminant of drinking water and food and feed chain. Nitrite induces oxidative damage in humans and animals. In this work, we studied the protective effect of crocin, the active constituent of Crocus sativus (saffron), on NaNO2 -induced oxidative damage in human erythrocytes. Changes in oxidative stress parameters following NaNO2 incubation of erythrocytes in presence and absence of crocin were determined. It was found that crocin pre-treatment significantly attenuated NaNO2 -induced oxidative damage of proteins, lipids, and plasma membrane. Crocin reduced the level of methemoglobin, the primary acute effect of nitrite intoxication. It also improved the antioxidant capacity of cells and NaNO2 -induced morphological changes in erythrocytes. Crocin is thus a potent protective agent against nitrite-induced cytotoxicity.

In vitro immunomodulatory effects of Astragalus verus Olivier. (black milkvetch): an immunological tapestry in Kurdish ethnomedicine

Astragalus verus Olivier., Fabaceae, has been used as an immunopromoting remedy in Kurdish ethnomedicine. Here, phagocytic (nitroblue tetrazolium (NBT) dye reduction and lysosomal enzyme activity of mouse splenocyte), (co)mitogenic (mouse splenocyte reduction of methyl thiazolyl tetrazolium (MTT)), and antioxidative (nitrite-induced methemoglobin formation) activities of A. verus extracts and chemical compositions of their essential oils (gas chromatography–mass spectrometry) were investigated. All aqueous (AQ), acetone (AC), and methanol (ME) extracts of aerial parts and root polysaccharide extract (PS) enhanced phagocytic activity compared with the control. The phagocytic activities assayed by lysosomal enzyme activity were in agreement with the values obtained by NBT reduction test and followed the order AQ > PS > AC > ME. All A. verus extracts showed enhanced mitogenic activities compared with the control; however, the AQ extract at 200 μg/mL showed maximum mitogenic activity by 311% compared with the control. The stimulation index values obtained by lipopolysaccharide comitogenic activity were roughly in agreement with the values obtained by concanavalin A comitogenic activity and followed the order AQ > PS > AC > ME. The AQ extract showed profound antioxidant activity while other extracts did not show an acceptable antioxidant activity. The main components isolated from the essential oil of A. verus were thymol in AQ, n-hexadecanoic acid, 1,2-benzenedicarboxylic acid, diisooctyl ester, and phytol in both ME and AC and tetradecane, pentadecane, hexadecane, and phenol,2,4-bis(1,1-dimethylethyl) in PS. The results clearly showed that A. verus could be employed as an immunopromoting remedy; however, more advanced researches are requested to develop a promised pharmaceutical drug from A. verus in the future.

HPLC analysis of human erythrocytic glutathione forms using OPA and N-acetyl-cysteine ethyl ester: Evidence for nitrite-induced GSH oxidation to GSSG

Glutathione exists in biological samples in the reduced form (GSH), as its disulfide (GSSG) and as a mixed disulfide (GSSR) with thiols (RSH). GSH is the most abundant low-molecular-mass thiol and plays important roles as a cofactor and as a main constituent of the intracellular redox status. Due to its own sulfhydryl (SH) group, GSH reacts readily with o-phthaldialdehyde (OPA) to form a highly stable and fluorescent isoindole derivative (GSH-OPA), which allows for sensitive and specific quantitative determination of GSH in biological systems by HPLC with fluorescence (FL) detection. In the present article we report on the utility of the novel, strongly disulfide bond-reducing thiol N-acetyl-cysteine ethyl ester (NACET) for the specific quantitative analysis of GSH and GSSG in the cytosol of red blood cells (RBC) as GSH-OPA derivative with FL (excitation/emission 338/458 nm) or UV absorbance (338 nm) detection. Unlike in aqueous solution, the derivatization of GSH in RBC cytosol yielded two closely related derivatives in the absence of NACET and only the GSH-OPA derivative in the presence of NACET. The HPLC method was optimized and validated for human RBC and applied to measure GSH and GSSG in RBC of healthy subjects. Basal GSH and GSSG concentrations were determined to be 2340 ± 350 μM and 11.4 ± 3.2 μM, respectively, in RBC of 12 healthy young volunteers (aged 23–38 years). The method was also applied to study the effects of nitrite on the glutathione status in intact and lysed human RBC. Nitrite at mM-concentrations caused instantaneous and considerable GSSG formation in lysed but much less pronounced in intact RBC. GSH externally added to lysed RBC inhibited nitrite-induced methemoglobin formation. Our findings suggest that nitric oxide/nitrite-related consumption rate of GSH, and presumably that of NADH and NADPH, could be of the order of 600 μmol/day in RBC of healthy subjects.

Anti-oxidant activity of ethyl acetate extract of <i> Aquilaria agallocha </i> on nitrite-induced methemoglobin formation

Anti-oxidant activity of ethyl acetate extract of <i> Aquilaria agallocha </i> on nitrite-induced methemoglobin formation

Effect of non steroidal anti-inflammatory drugs on inhibition of nitrite-induced methemoglobin formation

Paracetamol, Diclofenac, Aceclofenac, Nimesulide, Ibuprofen and Rofecoxib belonging to the class of non-steroidal anti-infl ammatory drugs (NSAID’s) were tested for inhibitory role on nitrite-induced oxidation of hemoglobin in vitro. Result indicates a promising inhibitory role of aceclofenac and signifi cant inhibitory role of ibuprofen in a concentration range of 0.25 mM each. Pro-oxidant activity was higher for remaining test compounds viz., Diclofenac, Paracetamol, Rofecoxib and Nimesulide. Key words: Free radical, methemoglobin, nitrite, NSAID

Effect of non steroidal anti-inflammatory drugs on inhibition of nitrite-induced methemoglobin formation

Effect of non steroidal anti-inflammatory drugs on inhibition of nitrite-induced methemoglobin formation

Nitrite uptake and metabolism and oxidant stress in human erythrocytes.

Nitric oxide, when released into the bloodstream, is quickly scavenged by Hb in erythrocytes or oxidized to nitrite. Nitrite can also enter erythrocytes and oxidize Hb. The goals of this work were to determine the mechanism of erythrocyte nitrite uptake and whether this uptake causes oxidant stress in these cells. Erythrocytes took up 0.8 mM nitrite with a half-time of 11 min. Nitrite uptake was sensitive to temperature and to the pH and ionic composition of the medium but was not inhibited by the specific anion-exchange inhibitor DIDS. About 25% of nitrite uptake occurred on the sodium-dependent phosphate transporter and the rest as diffusion of nitrous acid or other species across the plasma membrane. Methemoglobin formation increased in proportion to the intracellular nitrite concentration. Nitrite reacted with erythrocyte ascorbate, but ascorbate loading of cells decreased nitrite-induced methemoglobin formation only at high nitrite concentrations. In conclusion, nitrite rapidly enters erythrocytes and reacts with oxyhemoglobin but does not exert a strong oxidant stress on these cells.

Nitrite-induced methemoglobin formation and recovery in rainbow trout (Oncorhynchus mykiss) at high chloride concentrations

Nitrite-induced methemoglobin formation and recovery in rainbow trout (Oncorhynchus mykiss) at high chloride concentrations

Curcumin inhibits nitrite-induced methemoglobin formation

Curcumin protects hemoglobin from nitrite-induced oxidation to methemoglobin. The protection is not observed when curcumin is added after the autocatalytic stage of the oxidation of hemoglobin by nitrite. The ability of curcumin to scavenge superoxide may be responsible since superoxide is implicated in promoting the autocatalytic stage of oxidation of hemoglobin by nitrite.

Methemoglobin formation and reduction in canine erythrocytes with inherited high Na, K-ATPase activity.

Methemoglobin formation and reduction in canine erythrocytes with inherited high Na,K-ATPase activity (HK cells) were compared with those in normal canine cells (LK cells). Nitrite-induced methemoglobin formation in hemoglobin solutions indicated that the hemoglobin from HK cells was oxidized at essentially the same rate as that of LK cells. However, methemoglobin formation in HK cells was slower due to the inhibition by high glutathione (GSH) concentration. Methemoglobin reduction was allowed to take place on nitrite-treated and washed erythrocytes in a glucose medium and was reduced more rapidly in HK cells than in LK cells. During the reduction, the amounts of lactate and pyruvate increased more rapidly in HK cells, indicating enhanced glycolysis in HK cells. It is thus evident that the hemoglobin of HK cells is more securely protected from nitrite-induced oxidation by the GSH presence in great excess and by the increase in glycolysis.

Nitrite-induced methemoglobin formation and recovery in channel catfish (Ictalurus punctatus) at three acclimation temperatures.

The relationship between nitrite concentration and degree of methemoglobinemia has been documented in catfish; however, the influence of temperature on this process has not been examined. In light of the effects of temperature on the movement of chemicals, it is tempting to hypothesize that higher temperatures always increase toxicity. A temperature change may result in an increase, decrease or no change in the toxicity of a chemical pollutant. The authors conducted research to measure the effect of temperature on methemoglobin formation and recovery in nitrite-exposed channel catfish.

Dynamics of nitrite-induced methemoglobin formation after total gamma-ray irradiation of rats

One trend in the design of atomic power stations (APS) based on fast neutrons is the development of APS in which dissociating nitrogen tetroxide is used as heat carrier and as working heat. Here, the authors study the combined action of ionizing radiation and of sodium nitrite on methemoglobin formation associated with the action of nitrites as a metabolic product of nitrogen oxide metabolism. Experiments were carried out on male Wistar rats weighing 180-200 g. Single whole-body irradiation was given on the UGU-420 gamma-ray source in doses of 77.4 and 180.6 mCi/kg body weight. The dynamics of changes in the methemoglobin concentration was monitored on a VSU-2-P spectrophotometer at 540-630 nm. A diagram showing the relationship between nitrite-induced methemoglobin formation and dose and time elapsing after irradiation is shown.

The effect of ascorbic acid on nitrite-induced methemoglobin formation in rats, sheep, and normal human erythrocytes

Interspecies differences were demonstrated with respect to the occurrence of sodium nitrite-induced methemoglobin (METHB) in the erythrocytes of rats, dorset sheep, and normal humans, with the rats displaying approximately 25–33% of the sensitivity of sheep and human erythrocytes. Ascorbic acid incubation along with the nitrite was able to significantly reduce METHB formation in a dose-dependent manner in both rats and humans but not in sheep.

The effect of ascorbic acid on sodium nitrite-induced methemoglobin formation in glucose-6-phosphate dehydrogenase-deficient erythrocytes

Ascorbic acid significantly reduced the occurrence of sodium nitrite-induced methemoglobin (METHB) formation in a dose-dependent manner in erythrocytes from glucose-6-phosphate dehydrogenase (G-6-PD)-deficient humans in vitro. The ascorbic acid treatment, however, also decreased levels of reduced GSH in a dose-dependent manner, a response indicative of oxidant stress to the erythrocyte membrane. The latter findings are inconsistent with the hypothesis that ascorbic acid supplementation in G-6-PD-deficient humans may help compensate for inherently low levels of erythrocyte GSH. Finally, the ascorbic acid-induced reduction of METHB values, while of statistical significance, does not appear to be of clinical significance.

Low erythrocyte glucose-6-phosphate dehydrogenase (g-6-PD) activity and susceptibility to nitrite-induced methemoglobin formation.

Low erythrocyte glucose-6-phosphate dehydrogenase (g-6-PD) activity and susceptibility to nitrite-induced methemoglobin formation.

Nitrite-Induced Methemoglobin Formation in Channel Catfish

Nitrite that accumulates during intensive culture of channel catfish (Ictalurus punctatus) often results in high mortality. This study evaluated sublethal effects of nitrite on young channel catfish. Fish (7–10 cm standard length) exposed to 1, 2, 3, 4, or 5 mg/liter of nitrite for 24 hours developed mean methemoglobin concentrations (% of total hemoglobin) of 35, 79, 79, 85, and 90%, respectively. During the 5 mg/liter exposure, methemoglobin increased rapidly during the first 6 hours from 7% to 59%, more slowly to about 80% by 12 hours, and trivially thereafter. Channel catfish exposed to 5 mg/liter nitrite for 6 hours and transferred to nitrite-free water had near normal amounts of methemoglobin (13%) after 24 hours. An inverse relationship was noted between pH of aquarium water and methemoglobin formation after exposure to nitrite. Addition of calcium chloride, potassium chloride, sodium chloride, and sodium bicarbonate to the water suppressed methemoglobin formation in fish exposed to nitrite, but sodium sulfate had no such protective effect.

Chloride Inhibition of Nitrite-Induced Methemoglobinemia in Channel Catfish (Ictalurus punctatus)

Exposure of channel catfish (Ictalurus punctatus) fingerlings for 24 h to 1.0, 2.5, and 5.0 mg/L nitrite (pH = 7; hardness = 40 mg/L; temperature = 22–25 °C) produced methemoglobin levels of 20.7 ± 1.9%, 59.8 ± 1.9%, and 77.4 ± 1.4% (SE), respectively. However, methemoglobin levels were not elevated when fish were simultaneously exposed to 1.0, 2.5, and 5.0 mg/L nitrite and 25, 50, and 100 mg/L sodium chloride, respectively. Acclimation to sodium chloride for 24 h before exposure to nitrite did not enhance the inhibitory action of sodium chloride. Fish exposed to 5 mg/L nitrite for 5 h developed 42.5 ± 3.8% methemoglobin. When transferred to water containing 5 mg/L nitrite and 250 mg/L sodium chloride, methemoglobin levels returned to normal within 24 h. Environmental chloride probably inhibits methemoglobin formation by competing with nitrite for entrance into the gills of the fish. An ionic ratio of 16 Cl− to 1 NO2− is capable of complete suppression of nitrite-induced methemoglobin formation. Bicarbonate ion present in the test water (1 meq/L) may also have contributed to the inhibitive action of chloride. Key words: nitrite, methemoglobin, chloride, channel catfish

A rapid method for simultaneous measurement of carboxy- and methemoglobin in blood.

ArticleA rapid method for simultaneous measurement of carboxy- and methemoglobin in blood.K A Small, E P Radford, J M Frazier, F L Rodkey, and H A CollisonK A Small, E P Radford, J M Frazier, F L Rodkey, and H A CollisonPublished Online:01 Jul 1971https://doi.org/10.1152/jappl.1971.31.1.154MoreSectionsPDF (2 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformation Cited ByEndothelial Dysfunction in Passive Smokers23 December 2014 | Journal of Cardiology & Current Research, Vol. 1, No. 7Methods for the Detection of Gasotransmitters20 July 2012Are self-reports of smoking rate biased? Evidence from the Second National Health and Nutrition Examination SurveyJournal of Clinical Epidemiology, Vol. 48, No. 10Effects on the Heart of Two Forms of Chronic HypoxiaDiscrepancies between self-reported smoking and carboxyhemoglobin: an analysis of the second national health and nutrition survey.American Journal of Public Health, Vol. 82, No. 7Notice of Intended Change—Carbon Monoxide24 February 2011 | Applied Occupational and Environmental Hygiene, Vol. 6, No. 10Carboxyhemoglobin in the rat: Improvements in the spectrophotometric measurement and comparison to other studiesJournal of Applied Toxicology, Vol. 9, No. 5Chronic exposure to carbon monoxide at high altitude: Effects on mean electrical axisPhysiology & Behavior, Vol. 46, No. 1Arterial blood gas status in rats exposed to chronic CO at low and high altitudeRespiration Physiology, Vol. 75, No. 2Body and organ weights of rats exposed to carbon monoxide at high altitudeJournal of Toxicology and Environmental Health, Vol. 23, No. 3Serum Levels of Vitamin C in Relation to Dietary and Supplemental Intake of Vitamin C in Smokers and NonsmokersAnnals of the New York Academy of Sciences, Vol. 498, No. 1 Third ConfereDynamics of nitrite-induced methemoglobin formation after total gamma-ray irradiation of ratsBulletin of Experimental Biology and Medicine, Vol. 98, No. 5Prenatal Exposure to Carbon Monoxide: Learning and Memory DeficitsScience, Vol. 223, No. 4634Current Use of Ambient and Biological Monitoring: Reference Workplace Hazards. Inorganic Toxic Agents — Carbon Monoxide ICardiorespiratory responses to intestinal injection of carbon monoxidePharmacology Biochemistry and Behavior, Vol. 21Spectrophotometry of Hemoglobin and Hemoglobin DerivativesEffect of carbon monoxide on atherogenesis in normal pigs and pigs with von Willebrand's diseaseAtherosclerosis, Vol. 43, No. 2-3Measurement of smoking topography during withdrawal or deprivationBehavior Therapy, Vol. 12, No. 4A Multi-Wavelength Spectrophotometric Method for the Simultaneous Determination of Five Haemoglobin DerivativesClinical Chemistry and Laboratory Medicine, Vol. 19, No. 7Effects of prenatal carbon monoxide exposure on cardiac developmentToxicology and Applied Pharmacology, Vol. 56, No. 3Augmentation of Myocardial Ischemia by Low Level Carbon Monoxide Exposure in DogsArchives of Environmental Health: An International Journal, Vol. 34, No. 4A new method of carboxyhaemoglobin determination.Occupational and Environmental Medicine, Vol. 35, No. 1Lack of Effects of Carbon Monoxide on Human Vigilance6 October 2016 | Perceptual and Motor Skills, Vol. 45, No. 3Carboxyhaemoglobin. spectrophotometric determination tested and calibrated using a new reference method for measuring carbon monoxide in bloodClinica Chimica Acta, Vol. 80, No. 1Toxicity of Mild Prenatal Carbon Monoxide ExposureScience, Vol. 197, No. 4304Carbon monoxide and flash evoked potentials from rat cortex and superior colliculusPharmacology Biochemistry and Behavior, Vol. 6, No. 4The effects of brief exposures to carbon monoxide on temporally differentiated respondingEnvironmental Research, Vol. 12, No. 1Carbon Monoxide and Heart AttacksArchives of Environmental Health: An International Journal, Vol. 30, No. 10Hematological and biochemical effects of chronic co exposure on the Japanese quailRespiration Physiology, Vol. 24, No. 3Dichloromethane inhalation, carboxyhemoglobin concentrations, and drug metabolizing enzymes in rabbitsToxicology and Applied Pharmacology, Vol. 33, No. 3Effects of carboxyhemoglobin on the determination of methemoglobin in bloodBiochemical Medicine, Vol. 9, No. 3Comparison of three techniques for carboxyhaemoglobin determinationInternationales Archiv f�r Arbeitsmedizin, Vol. 33, No. 4Hemoglobin pigments. spectrophotometric determination of oxy-, carboxy-, met-, and sulfhemoglobin in capillary bloodClinica Chimica Acta, Vol. 42, No. 1A new method for the rapid determination of oxygen-dissociation curves in small blood samples by spectrophotometric titrationClinica Chimica Acta, Vol. 41 More from this issue > Volume 31Issue 1July 1971Pages 154-60 https://doi.org/10.1152/jappl.1971.31.1.154PubMed5556952History Published online 1 July 1971 Published in print 1 July 1971 Metrics

Hemoglobin M Reserve: studies on identification and characterization.

Abstract A family with hereditary cyanosis secondary to a hemoglobin M is reported; pending completion of amino acid analysis this abnormal hemoglobin is designated hemoglobin M Reserve . The M hemoglobin was separated by electrophoretic and chromatographic techniques and comprised 30 per cent of the proband's total hemoglobin. A qualitatively abnormal A 2 hemoglobin was also demonstrated. The isolated hemoglobin M had spectral absorption abnormalities in the 480 to 520 and 600 to 680 m μ ranges when in the oxy-, deoxy-, met-, and cyanmethemoglobin states; however, prolonged reduction with dithionite eliminated the spectral abnormalities present in the deoxygenated state. The structural abnormality of the hemoglobin M molecule was located in the alpha polypeptide chain and resulted in reduced oxygen affinity and decreased reversible oxygen binding capacity. Other demonstrable abnormalities included increased resistance to nitrite-induced methemoglobin formation, increased rate of ferricyanide-induced methemoglobin formation, and an increased rate of methemoglobin formation duing hemoglobin autooxidation. There were no demonstrable abnormalities in red cell content of reduced glutathione, glutathione peroxidase, TPN-methemoglobin reductase, or DPN diaphorase to explain these changes in rates of heme iron oxidation. The behavioral abnormalities are apparently secondary to the molecular conformational changes within the hemoglobin M molecule that are consequent upon the amino acid substitution, internal bonding of the ionized radical of the abnormal amino acid with heme iron, and altered interaction between alpha and beta polypepttde chains.