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It has been more than five years since the appearance of the first edition of Nitric Oxide Protocols. According to Medline, more than 32,500 publications dealing with nitric oxide have appeared in the primary literature in the int- vening time from 1999 to April 2004. These numbers attest to the continuing vitality of a field dealing with a deceptively simple, although extremely int- esting and promiscuous molecule, nitric oxide. Despite its apparent chemical simplicity, nitric oxide continues to amaze, with an extremely wide spectrum of functions and effects. In particular, the last few years have witnessed an explosion of nonclassical effects, involving the direct interaction of nitric - ide and related oxidized products with proteins to yield such novel combi- tions as nitrosylated and nitrated proteins. Another emergent area has been that of NO's clinical applications, including interesting developments related to the virus-mediated expression of nitric oxide synthase and the use of nitric oxide in gene therapy. In Nitric Oxide Protocols, I have gathered chapters dealing with the some of the latest methodologies for the measurement of nitric oxide.Many of these deal with quantitation in biological fluids or tissues, and they cover a wide spectrum of technologies, including capillary electrophoresis, microcoaxial electrodes, the in vivo measurement of nitric oxide in exhaled air, confocal microscopy, gas chromatography-mass spectroscopy, in situ hybridization, and real-time polymerase chain reaction.