PMID: 7542684Jul 17, 1995Paper

Measurement of methemoglobin formation from oxyhemoglobin. A real-time, continuous assay of nitric oxide release by human polymorphonuclear leukocytes

Journal of Immunological Methods
G Lärfars, H Gyllenhammar

Abstract

We have evaluated the spectrophotometric measurement (at 401 vs. 411 nm) of nitric oxide (NO)-dependent methemoglobin formation from oxyhemoglobin in order to assess NO release from human polymorphonuclear neutrophil leukocytes (PMN). S-nitroso-D,L-acetyl-penicillamine (SNAP, 25-200 microM), a donor of NO, induced a dose-dependent methemoglobin formation. Furthermore, when PMN were activated with N-formyl-methionylleucyl-phenylalanine or phorbol myristate acetate in the presence of superoxide dismutase (SOD) and catalase, methemoglobin formation ensued. The amount of methemoglobin formed was dependent on the amounts of oxyhemoglobin and stimulus used, and the number of PMN in the assay. The NO synthase (NOS) inhibitors NG-monomethyl-L-arginine or nitro-L-arginine methyl ester did not affect methemoglobin generation from oxyhemoglobin induced by SNAP but inhibited that mediated by activated PMN with IC50 values of 250 microM and 340 microM, respectively. The substrate for NO formation from NOS, L-arginine in concentrations up to 1 mM did not significantly influence the methemoglobin formation either induced by SNAP or activated PMN. Exclusion of SOD did not affect SNAP-dependent oxidation of oxyhemoglobin. Exclusion of SOD from ...Continue Reading

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Citations

Dec 8, 2000·Thrombosis Research·S Sethi, M Dikshit
Feb 13, 2001·ANZ Journal of Surgery·Y TsukaharaM Tanaka
Oct 16, 2012·Journal of Leukocyte Biology·Seymour J KlebanoffWilliam M Nauseef
Feb 13, 2015·Proceedings of the National Academy of Sciences of the United States of America·Errol L G SamuelJames M Tour
Mar 20, 2012·Nitric Oxide : Biology and Chemistry·Matthias TotzeckTienush Rassaf
Jul 17, 1998·The Journal of Laboratory and Clinical Medicine·G Lärfars, H Gyllenhammar
Jan 11, 2000·Circulation Research·S FukudaG W Schmid-Schönbein
May 21, 2019·Journal of Experimental Botany·Abhaypratap VishwakarmaKapuganti Jagadis Gupta
Aug 12, 2009·Analytica Chimica Acta·Marisa FreitasEduarda Fernandes

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