Mutagenic and carcinogenic potency of extracts of diesel and related environmental emissions: Study design, sample generation, collection, and preparation

A major diesel emissions research program has been initiated by the U.S. Environmental Protection Agency to assess the human health risk associated with increased use of diesel automobiles. This program is intended to establish the mutagenic and carcinogenic potency of complex organics associated with diesel particles as well as comparative particle-bound organics from other environmental emissions for which human epidemiological data are available. The mobile source samples selected for this study were collected from a heavy-duty diesel engine, a series of light-duty diesel passenger cars, and a gasoline catalyst automobile. The comparative source samples incorporated into the study were cigarette smoke condensate, coke oven emissions, roofing tar emissions, and benzo(a)pyrene. The samples were tested using three mutagenic assays and four carcinogenic assays as prescribed by a test matrix. This report describes the study design, particle generation, and sample collection and preparation. A brief summary of the bioassays is also included.     

The in-vitro response of pollen germination and tube length to different types of acidity

Pollen germination and tube growth are among the most sensitive responses to atmospheric pollution. Both these are inhibited by the acidity of the growth medium. Pollen grains from two species (Pinus cembra L. and Sambucus nigra L.) were germinated in media over a range of pHs (5.0, 4.5, 4.0, 3.5, 3.0 and 2.5) and six types of acidity (H(2)SO(4), HNO(3), H(2)SO(4): HNO(3) in 1:1, 2:1, 3:1 and 5:1 ratio). Pollen of the Gymnosperm is shown to be more resistant to acidity in the medium. Sulphuric acid alone and the ratio 2:1 with nitric acid are demonstrated to be the more harmful for P. cembra and S. nigra, respectively. The latter species was sensitive to all mixtures, particularly in respect to germination percentage.     

Metabolism of crufomate (4-tert-butyl-2-chlorophenyl methyl methylphosphoramidate) administered topically to a sheep.

A sheep dosed topically with 14C-crufomate (4-tert-butyl-2-chlorophenyl methyl methylphosphoramidate) excreted 45.5% of the 14C dose in the urine within 9 days. The feces contained 1.2% and the carcass 40.4% (this included the 37.7% of the dose remaining on the skin in the dosing area) of the dose. At sacrifice, the fat, liver, kidney, lung, and skin (where the dose was applied) contained the highest concentrations of 14C. Fourteen urinary metabolites were isolated and characterized by mass spectrometry. The metabolic reactions involved were oxidations of the t-butyl moiety, O-demethylation, replacement of the H-N-CH3 moiety with a hydroxyl group, oxidation of the N-methyl group to yield N-formyl phosphoramidates, hydrolysis of the phosphoramidate moiety to yield phenols, conjugation with glucuronic acid and combinations of these reactions.