Kenneth T. Bogen

Naphthalene metabolism in relation to target tissue anatomy, physiology, cytotoxicity and tumorigenic mechanism of action

This report provides a summary of deliberations conducted under the charge for members of Module C Panel participating in the Naphthalene State-of-the-Science Symposium (NS(3)), Monterey, CA, October 9-12, 2006. The panel was charged with reviewing the current state of knowledge and uncertainty about naphthalene metabolism in relation to anatomy, physiology and cytotoxicity in tissues observed to have elevated tumor incidence in these rodent bioassays. Major conclusions reached concerning scientific claims of high confidence were that: (1) rat nasal tumor occurrence was greatly enhanced, if not enabled, by adjacent, histologically related focal cellular proliferation; (2) elevated incidence of mouse lung tumors occurred at a concentration (30 ppm) cytotoxic to the same lung region at which tumors occurred, but not at a lower and less cytotoxic concentration (tumorigenesis NOAEL=10 ppm); (3) naphthalene cytotoxicity requires metabolic activation (unmetabolized naphthalene is not a proximate cause of observed toxicity or tumors); (4) there are clear regional and species differences in naphthalene bioactivation; and (5) target tissue anatomy and physiology is sufficiently well understood for rodents, non-human primates and humans to parameterize species-specific physiologically based pharmacokinetic (PBPK) models for nasal and lung effects. Critical areas of uncertainty requiring resolution to enable improved human cancer risk assessment were considered to be that: (1) cytotoxic naphthalene metabolites, their modes of cytotoxic action, and detailed low-dose dose-response need to be clarified, including in primate and human tissues, and neonatal tissues; (2) mouse, rat, and monkey inhalation studies are needed to better define in vivo naphthalene uptake and metabolism in the upper respiratory tract; (3) in vivo validation studies are needed for a PBPK model for monkeys exposed to naphthalene by inhalation, coupled to cytotoxicity studies referred to above; and (4) in vivo studies are needed to validate a human PBPK model for naphthalene. To address these uncertainties, the Panel proposed specific research studies that should be feasible to complete relatively promptly. Concerning residual uncertainty far less easy to resolve, the Panel concluded that environmental, non-cytotoxic exposure levels of naphthalene do not induce tumors at rates that can be predicted meaningfully by simple linear extrapolation from those observed in rodents chronically exposed to far greater, cytotoxic naphthalene concentrations.

Probabilistic Exposure Analysis for Chemical Risk Characterization

This paper summarizes the state of the science of probabilistic exposure assessment (PEA) as applied to chemical risk characterization. Current probabilistic risk analysis methods applied to PEA are reviewed. PEA within the context of risk-based decision making is discussed, including probabilistic treatment of related uncertainty, interindividual heterogeneity, and other sources of variability. Key examples of recent experience gained in assessing human exposures to chemicals in the environment, and other applications to chemical risk characterization and assessment, are presented. It is concluded that, although improvements continue to be made, existing methods suffice for effective application of PEA to support quantitative analyses of the risk of chemically induced toxicity that play an increasing role in key decision-making objectives involving health protection, triage, civil justice, and criminal justice. Different types of information required to apply PEA to these different decision contexts are identified, and specific PEA methods are highlighted that are best suited to exposure assessment in these separate contexts.

Efficient tumorigenesis by mutation-induced failure to terminate microRNA-mediated adaptive hyperplasia

Seven current contending cancer theories consider different sets of critical events as sufficient for tumorigenesis. These theories, most recently the microRNA dysregulation (MRD) theory, have overlapping attributes and extensive empirical support, but also some discrepancies, and some do not address both benign and malignant tumorigenesis. By definition, the most efficient tumorigenic pathways will dominate under conditions that selectively activate those pathways. The MRD theory provides a mechanistic basis to combine elements of the current theories into a new hypothesis that: (i) tumors arise most efficiently under stress that induces and sustains either protective or regenerative states of adaptive hyperplasia (AH) that normally are epigenetically maintained unless terminated; and (ii) if dysregulated by a somatic mutation that prevents normal termination, these two AH states can generate benign and malignant tumors, respectively. This hypothesis, but not multistage cancer theory, predicts that key participating AH-stem-cell populations expand markedly when triggered by stress, particularly chronic metabolic or oxidative stress, mechanical irritation, toxic exposure, wounding, inflammation, and/or infection. This hypothesis predicts that microRNA expression patterns in benign vs. malignant tumor tissue will correlate best with those governing protective vs. regenerative AH in that tissue, and that tumors arise most efficiently inmutagen-exposed stem cells that either happen to be in, or incidentally later become recruited into, an AH state.

Chamber for Testing Asbestos-Containing Products: Validation and Testing of a Re-created Chrysotile-Containing Joint Compound

Joint compound products containing chrysotile asbestos were commonly used for building construction from the late 1940s through the mid-1970s. Few relevant data exist to support reconstructing historical worker exposures to fibers generated by working with this material. Therefore, we re-created 1960s-era chrysotile-containing joint compound (JCC) and compared its characteristics to a current-day asbestos-free joint compound (JCN). Validation studies showed that a bench-scale chamber with controlled flow dynamics, designed to quantify particulate emissions from joint compound products, provided precise and reliable measurements of generated airborne dust mass, chrysotile fiber concentrations, and corresponding activity-specific emission rates. Subsequent chamber studies characterized fibers counted by phase contrast microscopy (PCM) per mass of respirable dusts and total suspended particulate dusts (total dusts), generated during JCC sanding or sweeping, as well as corresponding dust emission rates for JCC and JCN, and the ratio of total to respirable dust mass for JCN. From these data we estimated factors, F(CH-rd) and F(CH-td) (in units of f cm(-3) per mg m(-3)), by which respirable JCN dust mass concentrations collected during construction use can be converted to corresponding airborne PCM fiber concentrations generated by sanding or sweeping JCC. For sanding, median values (95% confidence limits) of F(CH-rd) and F(CH-td) were estimated to be 0.044 (0.039-0.050) and 0.212 (0.115-0.390) f cm(-3) per mg m(-3), respectively. The F(CH-td) to F(CH-rd) ratio indicates that approximately five times as many airborne PCM fibers are anticipated per unit air volume sampled when JCC dust is collected on cassettes (as done historically), than when respirable JCC dust is collected on cyclones. As the sizes of individual fibers collected appear to be primarily respirable, this difference may be a sampling artifact and suggests caution in interpreting historical fiber concentration measures made using cassettes during work with JCC-like materials. F(CH-rd) can be used with published and newly generated field measurements of respirable dust mass concentrations associated with the use of JCN or equivalent JCN materials to better characterize historical worker exposures to PCM fibers from use of JCC or equivalent JCCs. The experimental process described also can be used to develop conversion factors for other combinations of modern-day asbestos-free and historical chrysotile-containing products.

MECHANISTIC MODELS FIT TO ED001 DATA ON >40,000 TROUT EXPOSED TO DIBENZO[A,L]PYRENE INDICATE MUTATIONS DO NOT DRIVE INCREASED TUMOR RISK

ED001-study data on increased liver and stomach tumor risks in >40,000 trout fed dibenzo[a,l]pyrene (DBP), one of the most potently mutagenic chemical carcinogens known, provide the greatest low-dose dose-response resolution of any experimentally induced tumor data set to date. Although multistage somatic mutation/clonal-expansion cancer theory predicts that genotoxic carcinogens increase tumor risk in linear no-threshold proportion to dose at low doses, ED001 tumor data curiously exhibit substantial low-dose nonlinearity. To explore the role that nongenotoxic mechanisms may have played to yield such nonlinearity, the liver and stomach tumor data sets were each fit by two models that each assume a genotoxic and a nongenotoxic pathway to increased tumor risk: the stochastic 2-stage (MVK) cancer model, and a model implementing the more recent dysregulated adaptive hyperplasia (DAH) theory of tumorigenesis. MVK and DAH fits to the data sets were each excellent, but unexpectedly each MVK fit implies that DBP acts to increase tumor risk by entirely non-mutagenic mechanisms. Given that DBP is such a potent mutagen, the MVK-model fits obtained appear to be biologically implausible, whereas the DAH-model fits reflect that models assumption that chemical-induced tumorigenesis typically is driven by elevated repair-cell populations rather than mutations per se.

Reassessment of MTBE cancer potency considering modes of action for MTBE and its metabolites

Abstract A 1999 California state agency cancer potency (CP) evaluation of methyl tert-butyl ether (MTBE) assumed linear risk extrapolations from tumor data were plausible because of limited evidence that MTBE or its metabolites could damage DNA, and based such extrapolations on data from rat gavage and rat and mouse inhalation studies indicating elevated tumor rates in male rat kidney, male rat Leydig interstitial cells, and female rat leukemia/lymphomas. More recent data bearing on MTBE cancer potency include a rodent cancer bioassay of MTBE in drinking water; several new studies of MTBE genotoxicity; several similar evaluations of MTBE metabolites, formaldehyde, and tert-butyl alcohol or TBA; and updated evaluations of carcinogenic mode(s) of action (MOAs) of MTBE and MTBE metabolites. The lymphoma/leukemia data used in the California assessment were recently declared unreliable by the U.S. Environmental Protection Agency (EPA). Updated characterizations of MTBE CP, and its uncertainty, are currently needed to address a variety of decision goals concerning historical and current MTBE contamination. To this end, an extensive review of data sets bearing on MTBE and metabolite genotoxicity, cytotoxicity, and tumorigenicity was applied to reassess MTBE CP and related uncertainty in view of MOA considerations. Adopting the traditional approach that cytotoxicity-driven cancer MOAs are inoperative at very low, non-cytotoxic dose levels, it was determined that MTBE most likely does not increase cancer risk unless chronic exposures induce target-tissue toxicity, including in sensitive individuals. However, the corresponding expected (or plausible upper bound) CP for MTBE conditional on a hypothetical linear (e.g., genotoxic) MOA was estimated to be ∼2 × 10–5 (or 0.003) per mg MTBE per kg body weight per day for adults exposed chronically over a lifetime. Based on this conservative estimate of CP, if MTBE is carcinogenic to humans, it is among the weakest 10% of chemical carcinogens evaluated by EPA.

Potential Artifacts Associated with Historical Preparation of Joint Compound Samples and Reported Airborne Asbestos Concentrations

Airborne samples collected in the 1970s for drywall workers using asbestos-containing joint compounds were likely prepared and analyzed according to National Institute of Occupational Safety and Health Method P&CAM 239, the historical precursor to current Method 7400. Experimentation with a re-created, chrysotile-containing, carbonate-based joint compound suggested that analysis following sample preparation by the historical vs. current method produces different fiber counts, likely because of an interaction between the different clearing and mounting chemicals used and the carbonate-based joint compound matrix. Differences were also observed during analysis using Method 7402, depending on whether acetic acid/dimethylformamide or acetone was used during preparation to collapse the filter. Specifically, air samples of sanded chrysotile-containing joint compound prepared by the historical method yielded fiber counts significantly greater (average of 1.7-fold, 95% confidence interval: 1.5- to 2.0-fold) than those obtained by the current method. In addition, air samples prepared by Method 7402 using acetic acid/dimethylformamide yielded fiber counts that were greater (2.8-fold, 95% confidence interval: 2.5- to 3.2-fold) than those prepared by this method using acetone. These results indicated (1) there is an interaction between Method P&CAM 239 preparation chemicals and the carbonate-based joint compound matrix that reveals fibers that were previously bound in the matrix, and (2) the same appeared to be true for Method 7402 preparation chemicals acetic acid/dimethylformamide. This difference in fiber counts is the opposite of what has been reported historically for samples of relatively pure chrysotile dusts prepared using the same chemicals. This preparation artifact should be considered when interpreting historical air samples for drywall workers prepared by Method P&CAM 239.

Generalized Haber's Law for Exponential Concentration Decline, with Application to Riparian‐Aquatic Pesticide Ecotoxicity

A simple analytic solution to the dynamic version of Habers law was derived, conditional on a specified toxic load exponent (n) and on exponential decline in environmental toxicant concentration. Such conditions are particularly relevant to assessing ecotoxicity risk posed (e.g., to juvenile salmonids) by agricultural organophosphate (OP) pesticides that are subject to degradation and/or dissipation. A dynamic Habers law model was fit to previously published detailed data on lethality for two aquatic species induced by six agricultural OP pesticides, and more crude fits were obtained to less detailed data on five other OP and on two non-OP pesticides, indicating that for lethality, a range of 0.5 ≤ n ≤ 1.5 may be typical for OP pesticides. The AgDRIFT(®) stream deposition model was next used to establish that first-order or exponential loss, with dilution half-times on the order of ≤0.01 days, pertains approximately to pesticide residues in streams that arise after aerial application of agricultural pesticides 100 feet upwind. The analytic model was then applied to demonstrate that pesticide concentrations deposited in downwind streams following an aerial application are effectively diluted by about 50- to 300-fold from their initial concentration. Riparian ecotoxicity risk assessment models that ignore this effective dilution, and base pesticide-specific estimates of reduced survival on the initial concentrations, are therefore unrealistically conservative.

Generic Hockey-Stick Model for Estimating Benchmark Dose and Potency: Performance Relative to BMDS and Application to Anthraquinone

Benchmark Dose Model software (BMDS), developed by the U.S. Environmental Protection Agency, involves a growing suite of models and decision rules now widely applied to assess noncancer and cancer risk, yet its statistical performance has never been examined systematically. As typically applied, BMDS also ignores the possibility of reduced risk at low doses (“hormesis”). A simpler, proposed Generic Hockey-Stick (GHS) model also estimates benchmark dose and potency, and additionally characterizes and tests objectively for hormetic trend. Using 100 simulated dichotomous-data sets (5 dose groups, 50 animals/group), sampled from each of seven risk functions, GHS estimators performed about as well or better than BMDS estimators, and a surprising observation was that BMDS mis-specified all of six non-hormetic sampled risk functions most or all of the time. When applied to data on rodent tumors induced by the genotoxic chemical carcinogen anthraquinone (AQ), the GHS model yielded significantly negative estimates of net potency exhibited by the combined rodent data, suggesting that—consistent with the anti-leukemogenic properties of AQ and structurally similar quinones—environmental AQ exposures do not likely increase net cancer risk. In addition to its simplicity and flexibility, the GHS approach offers a unified, consistent approach to quantifying environmental chemical risk.

Exposures from chrysotile-containing joint compound: evaluation of new model relating respirable dust to fiber concentrations.

The potential for fiber exposure during historical use of chrysotile-containing joint compounds (JCC) has been documented, but the published data are of limited use for reconstructing exposures and assessing worker risk. Consequently, fiber concentration distributions for workers sanding JCC were independently derived by applying a recently developed model based on published dust measurements from sanding modern-day (asbestos-free) joint compound and compared to fiber concentration distributions based on limited historical measurements. This new procedure relies on factors that account for (i) differences in emission rates between modern-day and JCC and (ii) the number of fibers (quantified by phase contrast microscopy [PCM]) per mass of dust generated by sanding JCC, as determined in a bench-scale chamber study using a recreated JCC, that convert respirable dust concentrations to fiber concentrations. Airborne respirable PCM-fiber concentration medians (and 95% confidence intervals) derived for output variables using the new procedure were 0.26 (0.039, 1.7) f/cm(3) and 0.078 (0.013, 0.47) f/cm(3) , and corresponding total fiber concentrations were 1.2 (0.17, 9.2) f/cm(3) and 0.37 (0.056, 2.5) f/cm(3) , in enclosed and nonenclosed environments, respectively. Corresponding estimates of respirable and total PCM fiber concentrations measured historically during sanding of asbestos-containing joint compound-adjusted for differences between peak and time-weighted average (TWA) concentrations and documented analytical preparation and sampling artifacts-were 0.15 (0.019, 0.95) f/cm(3) and 0.86 (0.11, 5.4) f/cm(3) , respectively. The PCM-fiber concentration distributions estimated using the new procedure bound the distribution estimated from adjusted TWA historical fiber measurements, suggesting reasonable consistency of these estimates taking into account uncertainties addressed in this study.