Mutagenicity in lung of big Blue((R)) mice and induction of tandem-base substitutions in Salmonella by the air pollutant peroxyacetyl nitrate (PAN): predicted formation of intrastrand cross-links.

Peroxyacetyl nitrate (PAN) is a ubiquitous air pollutant formed from NO(2) reacting with acetoxy radicals generated from ambient aldehydes in the presence of sunlight and ozone. It contributes to eye irritation associated with photochemical smog and is present in most urban air. PAN was generated in a chamber containing open petri dishes of Salmonella TA100 (gas-phase exposure). After subtraction of the background mutation spectrum, the spectrum of PAN-induced mutants selected at 3.1-fold above the background mutant yield was 59% GC-->TA, 29% GC-->AT, 2% GC-->CG, and 10% multiple mutations - primarily GG-->TT tandem-base substitutions. Using computational molecular modeling methods, a mechanism was developed for producing this unusual tandem-base substitution. The mechanism depends on the protonation of PAN near the polyanionic DNA to release NO(2)(+) resulting in intrastrand dimer formation. Insertion of AA opposite the dimerized GG would account for the tandem GG-->TT transversions. Nose-only exposure of Big Blue((R)) mice to PAN at 78ppm (near the MTD) was mutagenic at the lacI gene in the lung (mutant frequency +/-S.E. of 6.16+/-0.58/10(5) for controls versus 8.24+/-0.30/10(5) for PAN, P=0.016). No tandem-base mutations were detected among the 40 lacI mutants sequenced. Dosimetry with 3H-PAN showed that 24h after exposure, 3.9% of the radiolabel was in the nasal tissue, and only 0.3% was in the lung. However, based on the molecular modeling considerations, the labeled portion of the molecule would not have been expected to have been bound covalently to DNA. Our results indicate that PAN is weakly mutagenic in the lungs of mice and in Salmonella and that PAN produces a unique signature mutation (a tandem GG-->TT transversion) in Salmonella that is likely due to a GG intrastrand cross-link. Thus, PAN may pose a mutagenic and possible carcinogenic risk to humans, especially at the high concentrations at which it is present in some urban environments.

Comparison of chemiluminescence and ultraviolet ozone monitor responses in the presence of humidity and photochemical pollutants.

The effect of water vapor and other pollutants on ozone monitoring instruments was investigated. Five UV-type and two chemiluminescence-type monitors were employed in this study. The results of the study indicate that in systems containing ozone, water vapor and zero air only, the UV-based monitors showed negligible effects due to humidity. On average, the UV monitors were within 0.5 percent of independently determined ozone values judged to be extremely accurate. The chemiluminescence-based monitors showed systematically higher readings than the UV monitors with added water vapor. The effect was found to be linear with water vapor concentration with an average positive deviation of 3.0 percent per percent H2O at 25 degrees C. For these measurement, ozone concentrations ranged from 85 to 320 ppbv and water concentrations from 1 to 3 percent (i.e., dew point temperatures from 9 to 24 degrees C). These results are largely in agreement with previous studies conducted to measure this interference, although the present study extends the range of water concentrations tested. Studies were also performed with a smog chamber with simulated polluted air (containing paraffinic, olefinic and aromatic hydrocarbon precursors) and varying relative humidities. Although the presence of water vapor did not appear to represent a substantial interference in these systems, a positive interference was observed with the UV monitors. This interference was likely a result of the presence of toluene and some of its aromatic photooxidation products (e.g., benzaldehyde), which can be partially removed from the reference stream by the ozone scrubber within the UV monitor. If the compound absorbs radiation at 254 nm, it is detected as ozone. However, when the results are scaled back to ambient concentrations of toluene and NO(x), the effect appears to be very minor (ca. 3 percent under the study conditions). It is concluded that under atmospheric conditions at moderate pollution and relative humidity levels, both types of instruments can give accurate measurements of the ozone concentration. These potential effects should be recognized when conducting ambient ozone measurements.

Comparison of mutagenic activities of several peroxyacyl nitrates.

Salmonella typhimurium, strain TA100 was exposed to a series of peroxyacyl nitrates including peroxyacetyl nitrate (PAN), peroxypropionyl nitrate (PPN), peroxybutyryl nitrate (PBN), peroxybenzoyl nitrate (PBzN), and chloroperoxyacetyl nitrate (CPAN). Gas-phase concentrations for the individual exposures were in the high part per billion by volume ppbv range. The dose was determined from the deposition rate and measured from the net decrease of the test compound in the exposure chamber and the exposure time. The mutagenic activity for each compound determined from the dose-response relationship gave values ranging from 250 (PBN) to 6,500 (PBzN) revertants/mumols. The mutagenic activity for CPAN could not be determined, due to an interference from chloroacetaldehyde. The difficulties of quantifying the actual gas-phase chemical dose the bacteria are exposed to in this variant of the Ames test are delineated.