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.     

Mutagenic and carcinogenic potency of extracts from diesel related environmental emissions: Simultaneous morphological transformation and mutagenesis in BALB/c 3t3 cells

Particulate extracts from six different environmental emission sources were assayed for genotoxic activity in mouse BALB/c 3T3 clone A31-1 cells. All compounds were tested simultaneously for both transforming and mutagenic (induction of ouabain-resistance) potential with and without exogenous metabolic activation in the form of a 9000 × g postmitochondrial hepatic supernatant fraction from Aroclor-1254 induced Fischer 344 rats. Dichloromethane particulate extracts from the exhaust of two light duty diesel engines (Oldsmobile and Nissan), one heavy duty diesel engine (Caterpillar) and one late model gasoline engine (Mustang II) were assayed in an identical manner to particulate extracts from the emissions of a roofing tar pot and a coke oven. No clear dose-dependent responses were observed, but several of the samples showed significant transforming and mutagenic activity. A qualitative ranking system showed the activity of these particulate extracts for either mutagenesis or transformation was: coke oven = Mustang II gasoline engine > Nissan diesel engine > roofing tar. Particulate extracts from the Oldsmobile diesel engine and the Caterpillar diesel engine showed essentially no activity.     

The additivity of multiple doses of a liver carcinogen in rats

The temporal pattern and dose-rate dependence of liver cancer in rats fed 2-fluorenylacetamide (2-FAA) was compared to analytical formulas derived from a specific model. The model was based on the idea that carcinogens have two distinct actions on cells: (1) transformation and (2) acceleration of progression. Transformed cells are by definition progressing toward cancer, but their rate of progression may depend on several factors, specifically, the amount of carcinogen. In the experiments the carcinogen was removed from the diet after various times and the rate of tumor progression was estimated from the time of tumor occurrence. The results were consistent with the idea that 2-FAA accelerated the progression of the cancers. The model provides a simple explanation of Druckrey's law in that the temporal occurrence of tumors induced by a carcinogen represents the acceleration of a naturally occurring disease progression.     

Plutonium in sheep faeces as an indicator of deposition on vegetation

In order to study the deposition on vegetation of plutonium arising from the testing of nuclear weapons and discharges from nuclear facilities, sheep faeces were collected from the South-west and North of England. The plutonium content of faeces has been shown to be a reproducible and sensitive indicator of deposition on herbage and, with this technique, plutonium discharged from the Sellafield Works of British Nuclear Fuels LImited can be detected 60 km from the site. On moorland pastures the concentration of ^239+240Pu in faeces, expressed as activity per gram of ash, is roughly equal numerically to the deposit of plutonium on 1 m² of vegetation.Some of the plutonium ingested by a grazing sheep is absorbed and the highest concentrations are found in liver. The liver of a sheep grazing on pasture where the ^239+240Pu concentration in faeces was about 2 pCi (74 mBq) g⁻¹ of ash, was found to contain <1% of the Generalised Derived Limit for mutton and offal, and the meat <0·001%.     

An elemental rationing technique for assessing concentration data from a complex water system

Water samples have been collected at the surface and bottom layers at 51 locations throughout Chesapeake Bay. The suspended particulate and dissolved fractions of these samples have been analyzed for Cd, Ce, Co, Cu, Fe, Mn, Mo, Ni, Pb, Sc, Sn, Th, U, and Zn using neutron activation analysis and atomic absorption spectrometry. Special chemical procedures were used to preconcentrate the elements of interest in the dissolved samples and separate them from the salt water matrix. The elemental concentrations observed in the dissolved samples were evaluated by direct comparison to those found in coastal seawater; however, the elemental concentrations in the particulate samples (mass per volume of water) were strongly influenced by the total amount of particulate material suspended in the water at time of collection. A double normalization procedure was used to calculate crustal enrichment factors for each sample, and these enrichment factors provided both a means to observe sample-to-sample variations, and also allowed a crude comparison with the natural levels occurring in the earth's crust.     

Threshold and dose response estimation using the “filter model”

The “filter model” has been developed to explain the biologic effects of radiation and chemicals. We have examined nearly 300 sets of dose response data, of which 50 are presented here. Responses (induced by radiation and chemicals) which have been examined include in vitro survival studies on animal and plant tissues, induction of cellular aberrations and time to tumor or death. Similar data from in vivo studies has also been examined. All of the data appear to fit the model R = a lnD + b(lnD)² + c, where R is the response, a and b are parameters fitted by regression to a particular set of data, and c is the response at zero (or lowest) dose. By writing this model in exponential form, it can be seen that the response R results from multistage filtering (by net amounts a and b) of the initial dose, D. The threshold is obtained from this model as the point, $\text{D}\text{?}{}_{\mathrm{T}}$, at which the second derivative becomes zero. This is given by $\text{D}\text{?}{}_{\mathrm{T}}\text{=}\text{exp}\text{(1 ?}\text{a}\text{2b}\text{)}$ when a and b are oppositelt signed.     

Fertility and air pollution: Some empirical findings

This paper attempts to explain regional differences in fertility rates in the standard metropolitan statistical areas of the United States through socioeconomic factors and air pollution. An important feature of this study is that it uses the most recent data pertaining to a homogeneous sample of observations within a country rather than using data for different countries. Statistical results suggest that given other factors, fertility and the level of air pollution have an inverse relationship. This quantitative effect is statistically significant but the absolute impact is rather low. Our results also suggest that an increase in the level of income, relative education of females, temperature, and the availability of physicians would in the long run reduce fertility rates in the urban areas of the United States.     

Health effects of oxidants

Oxidants of significance to human health include ozone, nitrogen dioxide, and peroxyacetylnitrate. All of these compounds are involved in complex photochemical reactions which makes quantification and prediction of their individual health effects difficult. Ozone causes trauma to lung tissues and interferes with enzyme systems in the lungs and other tissues causing a broad range of symptoms. Measurable health impacts can occur at concentrations as low as 390 μg/m³. Acute effects of ozone exposure are reversible at normal urban concentrations (80–120 μg/m³). A special problem of concern, however, is increased susceptibility to infectious diseases contracted through the lungs. Nitrogen dioxide also causes trauma to lung tissues and interferes with enzyme systems. Measurable impacts can occur at concentrations as low as 100 μg/m³, but recovery is rapid and it is not known whether repeated exposures at this level have cumulative effects or predispose the lungs to permanent damage. Chronic exposure of laboratory animals to higher nitrogen dioxide levels can cause emphysema-like conditions and reduction in resistance to respiratory infection. Epidemiological studies of children in houses with gas stoves confirm the finding of reduced resistance to respiratory infection. The U.S. EPA estimates that health effects may occur in young children exposed to concentrations in excess of 280 to 560 μg/m³ one-hour average. These concentrations occur routinely in houses having gas stoves. Peroxyacetylnitrate is a powerful eye irritant in photochemical smog. Other health effects are similar to those of ozone, but less important because of the relatively low concentrations of this pollutant compared to other oxidants.     

An appraisal of emissions control requirements in the California South Coast Air Basin

The purpose of this study is to evaluate the effect of reductions of reactive organic gases (ROG) and NO_x emissions on short-term O₃ and NO₂ concentrations and annual average NO₂ concentrations in the California South Coast Air Basin. Short-term air quality predictions were obtained by applying the Systems Applications Airshed Model to summer O₃ and autumn NO₂ episodes. Effects of emission controls on annual NO₂ concentrations were estimated using CDM and a new parcel tracking model NOXTRAK. Results for the summer O₃ episode indicate that ROG emission reduction in an effective means for reducing peak O₃ concentrations. NO_x emission reduction imposed in addition to ROG emission reductions are counterproductive in reducing peak O₃ concentrations. The modeling results also suggest that attainment of the 1-h federal O₃ standard requires ROG emission reductions on the order of 80% from 1987 levels. Results for the autumn NO₂ episode indicate that NO_x emission reductions approximating those recommended in a proposed Air Quality Management Plan (about 22%) will result in only small (about 5%) reductions in the peak NO₂ concentrations. ROG emission reduction may be more effective than NO_x emission reduction in reducing the peak NO₂ concentration. For the episode studied, a reduction of 36% in ROG emissions is estimated to result in a reduction in peak NO₂ concentrations commensurate with that required to attain the 1-h state NO₂ standard. Model calculations also indicate that the federal NO₂ standard may not be meet by 1987 at one or two stations, but may blosely approached.