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Fish Oil and Oxidative Stress by Inflammatory Leukocytes
Authors:Yvonne Mei Sian Tay  Kok Seong Lim  Fwu-Shan Sheu  Andrew Jenner  Matthew Whiteman  Kim Ping Wong
Institution:1. Department of Biochemistry, Faculty of Medicine, National University of Singapore, MD 7 #, 8 Medical Drive, 117597, Singapore, Singaporebchbh@nus.edu.sg;3. Department of Biochemistry, Faculty of Medicine, National University of Singapore, MD 7 #, 8 Medical Drive, 117597, Singapore, Singapore;4. Department of Biological Sciences, Faculty of Science, National University of Singapore, 14 Science Drive 4, 117543, Singapore, Singapore
Abstract:Several papers have claimed that mitochondria contain nitric oxide synthase (NOS) and make nitric oxide (NO?) in amounts sufficient to affect mitochondrial respiration. However, we found that the addition of l-arginine or the NOS inhibitor l-NMMA to intact rat liver mitochondria did not have any effect on the respiratory rate in both State 3 and State 4. We did not detect mitochondrial NO? production by the oxymyoglobin oxidation assay, or electrochemically using an NO? electrode. An apparent NO? production detected by the Griess assay was identified as an artifact. NO? generated by eNOS added to the mitochondria could easily be detected, although succinate-supplemented mitochondria appeared to consume NO?.

Our data show that NO? production by normal rat liver mitochondria cannot be detected in our laboratory, even though the levels of production claimed in the literature should easily have been measured by the techniques used. The implications for the putative mitochondrial NOS are discussed.
Keywords:Mitochondria  Nitric oxide  Nitric oxide synthase  Artifact  Griess assay
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