Use of a statistical model to predict the potential for repeated dose and developmental toxicity of dermally administered crude oil and relation to reproductive toxicity.

Petroleum (commonly called crude oil) is a complex substance primarily composed of hydrocarbon constituents. Based on the results of previous toxicological studies as well as occupational experience, the principal acute toxicological hazards are those associated with exposure by inhalation to volatile hydrocarbon constituents and hydrogen sulfide, and chronic hazards are associated with inhalation exposure to benzene and dermal exposure to polycyclic aromatic compounds. The current assessment was an attempt to characterize the potential for repeated dose and/or developmental effects of crude oils following dermal exposures and to generalize the conclusions across a broad range of crude oils from different sources. Statistical models were used to predict the potential for repeated dose and developmental toxicity from compositional information. The model predictions indicated that the empirical data from previously tested crude oils approximated a "worst case" situation, and that the data from previously tested crude oils could be used as a reasonable basis for characterizing the repeated dose and developmental toxicological hazards of crude oils in general.

Characterization of the noncancer hazards of gas oils.

Gas oils, used to manufacture diesel fuel and residential heating oil, are complex hydrocarbon substances with carbon numbers of C9-C30 and boiling ranges of approximately 150 °C to 450 °C. Target organ (liver enlargement, reduced thymus weights, and reductions in hematological parameters) and developmental (reduced fetal viability, increased resorption frequency, and reduced fetal weights) effects are associated with aromatic constituents present in some gas oils. Two types of gas oils were tested for repeated-dose and developmental toxicity following repeated dermal administration. A blend of commercial diesel fuels containing 26% aromatics, primarily single-ring compounds, did not cause either target organ or developmental effects at levels up to 600 mg/kg/d. "Cracked" gas oils containing higher levels of aromatic constituents were also tested. Because of limited sample availability, 2 cracked gas oil samples were tested, one for systemic effects and the other for developmental toxicity. The sample tested in the repeated-dose toxicity study (81% aromatics including approximately 10% 3-ring compounds) produced increased liver weights, reduced thymus weights, and reductions in hematological parameters. The overall no observed adverse effect level (NOAEL) was 100 mg/kg/d. The sample tested for developmental toxicity (65% aromatics including approximately 5% 3-ring compounds) resulted in significant reductions in fetal survival, significant increases in resorption frequency, and significant reductions in fetal weights with an overall NOAEL of 100 mg/kg/d. In summary, gas oils may or may not cause target organ and/or developmental effects depending on the levels and types of aromatic constituents that they contain.

Extreme sensitivity and the practical implications of risk assessment thresholds.

Traditional risk-assessment theory assumes the existence of a threshold for non-cancer health effects. However, a recent trend in environmental regulation rejects this assumption in favor of non-threshold linearity for these endpoints. This trend is driven largely by two related concepts: (1) a theoretical assumption of wide-ranging human sensitivity, and (2) inability to detect thresholds in epidemiologic models. Wide-ranging sensitivity assumes a subpopulation with extreme background vulnerability, so that even trivial environmental exposures are hazardous to someone somewhere. We use examples from the real world of clinical medicine to show that this theoretical assumption is inconsistent with the biology of mammalian systems and the realities of patient care. Using examples from particulate-matter air-pollution research, we further show that failure to reject linearity is usually driven by statistical rather than biological considerations, and that nonlinear/threshold models often have a similar or better fit than their linear counterparts. This evidence suggests the existence of practical, real-world thresholds for most chemical exposures.

The relationship between repeat-dose toxicity and aromatic-ring class profile of high-boiling petroleum substances.

A study was undertaken within the context of the U.S. EPA HPV Chemical Challenge Program to (1) characterize relationships between PAC content and repeat-dose toxicities of high-boiling petroleum substances (HBPS) and (2) develop statistical models that could be used to predict the repeat-dose toxicity of similar untested substances. The study evaluated 47 repeat-dose dermal toxicity and 157 chemical compositional studies. The four most sensitive endpoints of repeat-dose toxicity were platelet count, hemoglobin concentration, relative liver weight and thymus weight. Predictive models were developed for the dose-response relationships between the wt.% concentration of each of seven ring classes of aromatic compounds (the "ARC profile") and specific effects, with high correlations (r=0.91-0.94) between the observed and model-predicted data. The development of the mathematical models used to generate the results reported in this study is described by Nicolich et al. (2013). Model-generated dose-response curves permit the prediction of either the effect at a given dose or the dose that causes a given effect. The models generate values that are consistent with other standard measures. The models, using compositional data, can be used for predicting the repeat-dose toxicity of untested HBPS.

The relationship between developmental toxicity and aromatic-ring class profile of high-boiling petroleum substances.

In response to the US EPA HPV Challenge Program, this study was conducted to: (1) evaluate the relationship between PAC content and the developmental toxicity of high-boiling petroleum substances (HBPS) and (2) develop mathematical models to predict the developmental toxicity of similar untested substances based on their aromatic ring class (ARC) profiles. For this investigation, 68 developmental toxicity studies were reviewed. The ARC models relied on data from 21 rat dermal developmental toxicity studies conducted with similar experimental designs to ensure a consistent data set for comparison. The most sensitive general endpoints of developmental toxicity (i.e., decreased fetal survival and growth) were chosen for modeling. The ARC models demonstrated a strong correlation between the predicted vs. observed values for specific sensitive endpoints of these developmental toxicities (percent resorptions, r=0.99; live fetuses per litter, r=0.98; fetal body weight, r=0.94). Such associations provide a promising approach for predicting the developmental toxicity of untested HBPS. Efforts to corroborate the ARC models using test substances that were not used to build the ARC models produced mixed results, and further development and refinement of the ARC models is recommended before they can be reliably applied to all HBPS.

Genetic toxicity of high-boiling petroleum substances.

There are several specific types of high-boiling petroleum substances (HBPS) having final boiling points >343°C), in which genetic toxicity can be related to the content of polycyclic aromatic compounds (PACs), specifically crude oils, gas oils, heavy fuel oils, lubricant base oils, waxes and aromatic extracts. Evaluation of optimized Salmonella tests covering over 250 samples from 43 types of HBPS revealed that gene mutation can be determined for these substances using a protocol optimized for the detection of mutagenic PAC. The outcomes of modified Salmonella assays can be predicted using HBPS compositional information as input to a newly developed statistical model. The general outcome of the optimized Salmonella assay can be predicted for an untested substance based on its Aromatic Ring Class (ARC) profile. Review of the results from numerous cytogenetic tests showed that although a few positive study results have been reported, most HBPS do not produce chromosomal effects when tested in rodent bone marrow assays or in in vitro chromosomal aberration assays. Results of both bacterial and cytogenetic studies can be used to satisfy genetic toxicity endpoints for the HBPS category substances.

Evaluating the male and female reproductive toxicity of high-boiling petroleum substances.

To meet the EPA HPV Chemical Challenge Program requirement for reproductive toxicity data on sponsored high-boiling petroleum substances (HBPS), an analysis was conducted using the results of 39 repeat-dose and 59 developmental rat dermal toxicity studies on HBPS samples spanning the boiling range of the sponsored substances, and the results of three one-generation reproductive toxicity studies on two samples spanning the concentration range of polycyclic aromatic compounds of sponsored substances. The analysis found little evidence of male or female reproductive tract toxicity based on histopathology, reproductive organ weight, and sperm parameters, and no evidence of effects on fertility, while significant developmental toxicity and/or systemic repeat-dose toxicity were frequently observed. Among 14 samples of HBPS tested in both repeat-dose toxicity and developmental toxicity studies, there were no studies in which an adverse reproductive tract finding occurred at a dose lower than that producing developmental toxicity or other adverse effects in repeat-dose toxicity studies. The current analysis supports the hypothesis that effects in developmental and/or repeat-dose toxicity studies of HBPS occur at doses lower than those that might affect fertility in rat one-generation reproductive studies. When adequate developmental and repeat-dose toxicity studies are available, a reproductive toxicity study of HBPS appears unnecessary.

The development of statistical models to determine the relationship between aromatic-ring class profile and repeat-dose and developmental toxicities of high-boiling petroleum substances.

The repeat-dose and developmental toxicities of certain petroleum refinery streams are related to their polycyclic aromatic compound (PAC) content (Feuston et al., 1994). Building on this foundation, and working within the context of the US EPA High Production Volume (HPV) Chemical Challenge Program, we: (1) characterized relationships between PAC content and repeat-dose and developmental toxicities of high boiling petroleum substances (HBPS), and (2) developed statistical models that can be used to predict critical effects of similar untested substances. Data from 39 dermal toxicity studies of HBPS were used to develop statistical models to predict the dose-response relationships between the weight percent concentration of each of their 1-7 aromatic ring classes and 4 repeat-dose and 3 developmental endpoints (absolute thymus weight, hemoglobin count, platelet count, liver to body weight, live fetus count, fetal weight, and percent resorptions). The correlations between the observed and model-predicted values are >0.90. The predictive ability of the models was tested via a series of evaluation or corroboration methods. As is shown in the paper, using only compositional data of untested HBPS, the models can be used to predict the effect at a given dose or the dose that causes an effect of a stipulated magnitude.

Assessing the mammalian toxicity of high-boiling petroleum substances under the rubric of the HPV program.

In 1998, the US EPA announced the HPV Challenge Program, a voluntary chemical data collection effort. The Petroleum HPV Testing Group (PHPVTG(1)) volunteered to provide data on approximately 110 high boiling petroleum substances (HBPS), i.e. substances with final boiling points ≥ approximately 650°F (343°C). These HBPS are substances of unknown and variable composition (UVCBs) that are composed of numerous individual constituents. Toxicity studies have shown that some HBPS can produce systemic (repeat-dose) and developmental effects, and some are mutagenic under in vitro conditions. The papers in this supplement show that these effects are related to the profiles of aromatic constituents in these substances. Further, it is shown that the effects on selected repeat-dose and developmental toxicity endpoints and mutagenic activity in bacterial assays can be predicted from compositional information using models based on the aromatic-ring class profile, "ARC profile" as defined by gas chromatographic separation of the DMSO-soluble fraction of the starting materials. This chromatographic method and the predictive models provide an efficient means of characterizing for screening purposes the potential for repeat-dose, developmental effects and bacterial mutagenicity of HBPS and can reduce the number of animal tests that would be required if these tests were conducted on all 110 HBPS.

Lung cancer incidence in Canadian petroleum workers.

OBJECTIVES: This study's purpose was to conduct a more in-depth analysis of the potential association between lung cancer, occupational exposures and smoking using data on cohort members from a Canadian petroleum company and refined statistical analyses. METHODS: Information on various exposures including asbestos and petroleum coke dust, as well as job type and operating segment were collected via manual and computerised company records. We performed life-table analyses, Poisson regression and restricted cubic splines to model exposure-response patterns while controlling for smoking status and age. Model diagnostics included the assessment of dispersion and offset parameters. RESULTS: These analyses show that lung cancer risk is strongly related to age and smoking, and to a lesser extent to province of last residence. When controlling for these covariates, there is suggestive evidence that maintenance work may also be related to lung cancer risk. Some analyses also indicate that asbestos exposure may be associated with lung cancer risk, although a clear exposure-response trend is not seen. Other exposures, including petroleum coke dust, were not strongly related to lung cancer risk, particularly when expressed as a continuous measure. CONCLUSIONS: These data suggest that maintenance work may be associated with lung cancer incidence, although exposures to the single agents studied did not emerge as strong predictors of lung cancer incidence. Maintenance work may be a surrogate for general exposures to several agents (eg, polycyclic aromatic hydrocarbons, metals, welding fumes, radiation, etc), although these results may be affected by residual confounding due to smoking or other socio-demographic factors.

Lung cancer and diesel exhaust: an updated critical review of the occupational epidemiology literature.

A recent review concluded that the evidence from epidemiology studies was indeterminate and that additional studies were required to support the diesel exhaust-lung cancer hypothesis. This updated review includes seven recent studies. Two population-based studies concluded that significant exposure-response (E-R) trends between cumulative diesel exhaust and lung cancer were unlikely to be entirely explained by bias or confounding. Those studies have quality data on life-style risk factors, but do not allow definitive conclusions because of inconsistent E-R trends, qualitative exposure estimates and exposure misclassification (insufficient latency based on job title), and selection bias from low participation rates. Non-definitive results are consistent with the larger body of population studies. An NCI/NIOSH cohort mortality and nested case-control study of non-metal miners have some surrogate-based quantitative diesel exposure estimates (including highest exposure measured as respirable elemental carbon (REC) in the workplace) and smoking histories. The authors concluded that diesel exhaust may cause lung cancer. Nonetheless, the results are non-definitive because the conclusions are based on E-R patterns where high exposures were deleted to achieve significant results, where a posteriori adjustments were made to augment results, and where inappropriate adjustments were made for the "negative confounding" effects of smoking even though current smoking was not associated with diesel exposure and therefore could not be a confounder. Three cohort studies of bus drivers and truck drivers are in effect air pollution studies without estimates of diesel exhaust exposure and so are not sufficient for assessing the lung cancer-diesel exhaust hypothesis. Results from all occupational cohort studies with quantitative estimates of exposure have limitations, including weak and inconsistent E-R associations that could be explained by bias, confounding or chance, exposure misclassification, and often inadequate latency. In sum, the weight of evidence is considered inadequate to confirm the diesel-lung cancer hypothesis.

A weight of evidence approach for selecting exposure biomarkers for biomonitoring.

CONTEXT: It is known that there are usually several biomarkers and/or medium combinations that can be applied to answer a specific exposure question. To help determine an appropriate combination for the specific question, we have developed a weight-of-evidence Framework that provides a relative appropriateness score for competing combinations. METHODS: The Framework is based on an expert assessor's evaluation of the relevance and suitability of the biomarker and medium for the question based on a set of criteria. We provide a computer based modeling tool to guide the researcher through the process. RESULTS: We present an example with six biomarkers of benzene exposure in one matrix; the six are either the most commonly used biomarkers and/or have recent widespread usage. The example clearly demonstrates the usefulness of the Framework for scoring the choices, as well as the transparency of the method that provides the basis for discussion. CONCLUSIONS: The Framework provides for the first time a method to transparently document the rationale behind selecting, from among a set of alternatives, the most scientifically supportable exposure biomarker to address a specific biomonitoring question, thus providing a reproducible account of expert opinions on the suitability of a biomarker.

Mortality patterns and trends among 49,705 U.S.-based women in a petroleum company: update 1979-2000.

OBJECTIVE: To examine mortality patterns and trends in a cohort of women employed in U.S. operating segments of a petroleum company. METHODS: Based on human resources databases, we defined a cohort of 49,705 U.S.-based women with at least one day of company employment during 1979 to 2000. These data sources provided demographic and most work history information. Standardized mortality ratios and 95% confidence intervals were calculated for 95 causes of death for the total cohort and with separate analyses by job type and operating segment when numbers allowed. RESULTS: Cohort women have a 25% lower overall death rate than the general U.S. female population comparison. This lower rate is expected in light of the "healthy worker effect" that influences employee studies. Circulatory diseases have a deficit of 40%, and external causes of death and cancer have deficits of 13% and 9%, respectively. For analyses by job type, office/clerical workers have an elevation in ovarian cancer (standardized mortality ratio = 1.40, 95% confidence interval = 1.02 to 1.87), based on 46 deaths, with no work-related patterns. White-collar groups have generally large overall deficits for noncancer causes of death. In contrast, and based on smaller numbers, operators and laborers have elevations of motor vehicle accidents and other external causes of death, and laborers also have elevations of cerebrovascular disease and chronic obstructive pulmonary disease. These variations by job type are probably associated with differences in lifestyle factors. CONCLUSIONS: This large mortality surveillance study of women in the petroleum industry provides an opportunity for meaningful analysis of many causes of death. The study found an overall favorable mortality profile and, for a small number of elevations, helped identify possible subgroups for health and safety prevention programs and interventions.

Peripheral blood effects in benzene-exposed workers.

The hematotoxic effects of benzene exposure may be important in the occurrence of subsequent health effects. We sought to provide further information on peripheral blood effects by studying 928 workers in five factories in and around Shanghai, China exposed to a wide range of benzene concentrations. Specifically, we sought to investigate which blood indices are more strongly related to benzene exposure and which concentration levels of benzene result in peripheral blood changes. Lifestyle habits and demographic information was obtained via questionnaire, and potentially important genetic influences were determined by assessing single nucleotide polymorphisms in four genes (NQO1, MPO, CYP2E1, GSTT1). Weekly benzene exposure estimated from individual monitoring results ranged from 0.07 to 872 mg/m(3) with a median value of 7.4 mg/m(3). Twelve peripheral blood indices were examined. Stronger effects on peripheral blood were seen for red cell indices such as anemia and macrocytosis, albeit at higher (>10 ppm) exposure levels. The most sensitive parameters to benzene appeared to be neutrophils and the mean platelet volume (MPV), where effects were seen for benzene air concentrations of 7.8-8.2 ppm. Toluene exposure is a potential confounder for some peripheral blood effects, pointing to the need to scrutinize levels of both compounds in the occupational environment.

Mortality patterns and trends among 127,266 U.S.-based men in a petroleum company: update 1979-2000.

OBJECTIVE: To assess patterns and trends in mortality among men employed in U.S. operating segments of a petroleum company. METHODS: We defined a cohort of 127,266 men with at least 1 day of employment during the period of 1979 through 2000. Computerized human resources databases were the basis of the cohort definition as well as the source of demographic and most work history information. Standardized mortality ratios (SMR) and 95% confidence intervals (CI) were calculated for 94 causes of death, including analyses by operating segment and job type. RESULTS: Most SMR results are below unity. The main exception is mesothelioma (SMR = 1.49; 95% CI = 1.15 to 1.90), which has elevations three times greater than expectation among some groups of men working in manufacturing sites who were hired before 1960. SMRs for cancers of the blood and blood-forming organs are generally close to unity, whereas men in the chemicals segment have 17 deaths due to acute non-lymphocytic leukemia (SMR = 1.81; 95% CI = 1.06 to 2.90), with no temporal or job type patterns. Men in the downstream segment have an elevation of aplastic anemia (SMR = 2.19; 95% CI = 0.95 to 4.32), based on eight deaths. There are eight deaths from malignant melanoma among downstream drivers (SMR = 2.46; 95% CI = 1.06 to 4.84), and motor vehicle accident rates are slightly elevated among some groups of younger and shorter-term operators. CONCLUSIONS: This comprehensive study indicates an overall favorable mortality profile for this workforce. For a few elevations, the study helps guide decisions about future surveillance, focused studies, and other follow-up actions.

Cumulative effects and threshold levels in air pollution mortality: data analysis of nine large US cities using the NMMAPS dataset.

We examined the existence of thresholds, cumulative effects and the homogeneity of five air pollutants on the relative risk of three mortality outcomes using data from nine major US cities using data from NMMAPS. Overall, PM(10) (usually 200-day accumulation) and ozone (3-day accumulation) were the two important predictors of outcome but their effect was not uniform across the nine cities. Many models exhibited thresholds (25-45 microm g/m(3) for PM(10), and 10-45 ppb for O(3)). Our preliminary exploratory analyses suggest that the use of a linear, no threshold, model for pollution studies is not consistent with the observed data. The heterogeneity in the risk estimates across the nine cities suggests combining the local risk estimates to obtain a national risk estimate may not be justifiable and the estimate is likely to be confounded.

Genotoxicity of intermittent co-exposure to benzene and toluene in male CD-1 mice.

Benzene is an important industrial chemical. At certain levels, benzene has been found to produce aplastic anemia, pancytopenia, myeloblastic anemia and genotoxic effects in humans. Metabolism by cytochrome P450 monooxygenases and myeloperoxidase to hydroquinone, phenol, and other metabolites contributes to benzene toxicity. Other xenobiotic substrates for cytochrome P450 can alter benzene metabolism. At high concentrations, toluene has been shown to inhibit benzene metabolism and benzene-induced toxicities. The present study investigated the genotoxicity of exposure to benzene and toluene at lower and intermittent co-exposures. Mice were exposed via whole-body inhalation for 6h/day for 8 days (over a 15-day time period) to air, 50 ppm benzene, 100 ppm toluene, 50 ppm benzene and 50 ppm toluene, or 50 ppm benzene and 100 ppm toluene. Mice exposed to 50 ppm benzene exhibited an increased frequency (2.4-fold) of micronucleated polychromatic erythrocytes (PCE) and increased levels of urinary metabolites (t,t-muconic acid, hydroquinone, and s-phenylmercapturic acid) vs. air-exposed controls. Benzene co-exposure with 100 ppm toluene resulted in similar urinary metabolite levels but a 3.7-fold increase in frequency of micronucleated PCE. Benzene co-exposure with 50 ppm toluene resulted in a similar elevation of micronuclei frequency as with 100 ppm toluene which did not differ significantly from 50 ppm benzene exposure alone. Both co-exposures - 50 ppm benzene with 50 or 100 ppm toluene - resulted in significantly elevated CYP2E1 activities that did not occur following benzene or toluene exposure alone. Whole blood glutathione (GSH) levels were similarly decreased following exposure to 50 ppm benzene and/or 100 ppm toluene, while co-exposure to 50 ppm benzene and 100 ppm toluene significantly decreased GSSG levels and increased the GSH/GSSG ratio. The higher frequency of micronucleated PCE following benzene and toluene co-exposure when compared with mice exposed to benzene or toluene alone suggests that, at the doses used in this study, toluene can enhance benzene-induced clastogenic or aneugenic bone marrow injury. These findings exemplify the importance of studying the effects of binary chemical interactions in animals exposed to lower exposure concentrations of benzene and toluene on benzene metabolism and clastogenicity. The relevance of these data on interactions for humans exposed at low benzene concentrations can be best assessed only when the mechanism of interaction is understood at a quantitative level and incorporated within a biologically based modeling framework.

Field Test of an Epidemiology Curriculum for Middle School Students.

The purpose of this study was to test the effectiveness of a middle school epidemiology curriculum called Detectives in the Classroom. The curriculum presents epidemiology as the science of public health, using health-related issues that capture the interest of young students and help prepare them to make evidence-based health-related decisions. The curriculum was field tested among seventh-grade urban students using a quasi-experimental design. Analysis of covariance of pre- and post-test scores examined five outcomes, including students' perceptions of their abilities in science as inquiry, scientific literacy, and knowledge about five enduring epidemiologic understandings; their self reported interest in science, and assessment of students' epidemiological reasoning ability. The 378 experimental students, compared to 620 controls, had generally higher post-test improvements in epidemiology-related outcomes and smaller increases in the other measures. A dose-response was suggested by higher scores among students exposed to more than 10 lessons. Strengths of this evaluation include a large sample and availability of data to account for differences in demographic and school performance variables. Limitations of this evaluation include randomization by school as opposed to student, the relatively short-term and generally self-reported outcomes, and inconsistencies in proportion of the curriculum actually taught. The findings offer encouragement about the potential for Detectives in the Classroom to improve students' perceptions of their science abilities and scientific literacy, their interest in science and their abilities in basic epidemiologic reasoning. Further tests of this and other epidemiology curricula are needed to respond to the growing interest in teaching public health science to younger students. And while it is important to test near-term impacts, an additional challenge from a curriculum evaluation standpoint will be to follow students over several years to examine subsequent choices concerning selected courses, college majors, and career paths.

Relationship between silicosis and lung function.

In an examination of the relationship between silicosis and lung function, relevant studies of silica-exposed workers were reviewed. Smoking, dust exposure, and emphysema are three important factors that can confound the association between silicosis and lung function. Despite the importance of smoking in relation to lung function, some studies did not control for smoking, or smoking was controlled inadequately. The data suggest a weak association between lung function (mainly obstruction) and dust exposure, although some studies had crude measures of exposure. In general, the lung function of those with radiographic silicosis in category 1 was indistinguishable from those in category 0. Those in category 2 had small reductions in lung function relative to those with category 0 and little difference in the prevalence of emphysema. There were slightly greater decrements in lung function with category 3 and more significant reductions with progressive massive fibrosis. Emphysema was related to higher categories of silicosis, as well as to smoking. Silica exposure was often inadequately controlled in studies examining silicosis and lung function. A few studies suggested that emphysema is an independent risk factor associated with significant reductions in lung function.