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This study shows conclusively that some of the direct microbial mutagenic activity of the soluble-organie-fraction from Diesel particulate matter can be attributed to 1-nitropyrene. 1-nitropyrene has been shown to be formed by the nitration of pyrene, and pyrene is one inherent product of the diffusion-controlled-combustion of hycrocarbons that occurs with Diesel engine operation. Nitrogen dioxide, in the presence of water vapor, is shown to be a potential nitrating agent, and this gas can be produced by the high temperature oxidation of the nitrogen contained in the oxidant. These results are based on studies which used a well-documented engine, model fuel, model oxidants, and synthetic lubricant.