Program specification: a precursor to program monitoring and quality improvement. A case study from Boysville of Michigan

The reaction of synthetic DOPA melanin (DM) with lactoperoxidase (LPO), hydrogen peroxide, and nitrite (NO2-) has been investigated using EPR. We observed that in the presence of nitrite LPO/H2O2 generated large amount of melanin radicals, as evidenced by a strong, up to 11-fold, increase in the intensity of the melanin EPR signal. In contrast, when nitrite was omitted the increase was much less, ca. 30%, which, nevertheless, indicates that DM can be metabolized directly by LPO/H2O2. When the nitrite was present, the concentration of melanin radicals was linearly dependent on [NO2-] (for [NO2-] <5 mM), and increased when [LPO] and [H2O2] increased (at constant [NO2-]). We propose that the mechanism for the generation of melanin radicals by the LPO/H2O2/nitrite system involves oxidation of NO2- by LPO/H2O2 to a reactive metabolite, most likely the nitrogen dioxide radical (.NO2), which subsequently reacts with melanin 5,6-dihydroxyindole subunits producing the respective semiquinone radicals. Because melanin and .NO2 generating systems (nitrite, peroxidase enzymes, hydrogen peroxide) may coexist in cells in vivo, our results suggest that melanin could function as a natural scavenger of this highly reactive nitrogen species. This property may be relevant to the physiological functions of the melanin pigments in vivo.