Quantitative structure-mesothelioma potency model optimization for complex mixtures of elongated particles in rat pleura: A retrospective study.

Cancer potencies of mineral and synthetic elongated particle mixtures, including asbestos fibers, are influenced by changes in fiber dose composition, bioavailability, and biodurability in combination with relevant cytotoxic dose-response relationships. An extensive rat intrapleural dose characterization data set with a wide variety of elongated particles physicochemical properties facilitated statistical analyses of pleural mesothelioma response data combined from several studies for evaluation of alternative dose-response models. Utilizing logistic regression of individual elongated particle dimensional variations within each test sample, four major findings emerged: (1) Mild acid leaching provides superior prediction of tumor incidence compared to samples that were not leached; (2) sum of the elongated particle surface areas from mildly acid-leached samples provides the optimum holistic dose-response model; (3) progressive removal of dose associated with very short and/or thin elongated particles significantly degrades the resultant particle count and surface area dose-based predictive model fits; and (4) alternative biologically plausible model adjustments provide evidence for reduced potency of elongated particles with aspect ratios less than 8 and lengths greater than 80 µm. Regardless of these adjustments, the optimum predictive models strongly incorporate potency attributable to abundant short elongated particles in proportion to their surface area. Transmission electron microscopy analyses of low-temperature-ashed pleural membrane and lung tissues 5.5 mo post intrapleural exposures do not support hypotheses that short elongated particles that reach the pleural space are rapidly eliminated. Low-aspect-ratio elongated particles were still abundant in pleural membrane tissues but may have reduced potencies due to aggregation tendencies and therefore lower potential for intracellular presence.

In vitro determinants of asbestos fiber toxicity: effect on the relative toxicity of Libby amphibole in primary human airway epithelial cells.

BACKGROUND: An abnormally high incidence of lung disease has been observed in the residents of Libby, Montana, which has been attributed to occupational and environmental exposure to fibrous amphiboles originating from a nearby contaminated vermiculite mine. The composition of Libby amphibole (LA) is complex and minimal toxicity data are available. In this study, we conduct a comparative particle toxicity analysis of LA compared with standard reference asbestiform amphibole samples. METHODS: Primary human airway epithelial cells (HAEC) were exposed to two different LA samples as well as standard amphibole reference samples. Analysis of the samples included a complete particle size distribution analysis, calculation of surface area by electron microscopy and by gas adsorption and quantification of surface-conjugated iron and hydroxyl radical production by the fibers. Interleukin-8 mRNA levels were quantified by qRT-PCR to measure relative pro-inflammatory response induced in HAEC in response to amphibole fiber exposure. The relative contribution of key physicochemical determinants on the observed pro-inflammatory response were also evaluated. RESULTS: The RTI amosite reference sample contained the longest fibers and demonstrated the greatest potency at increasing IL-8 transcript levels when evaluated on an equal mass basis. The two LA samples and the UICC amosite reference sample consisted of similar particle numbers per milligram as well as similar particle size distributions and induced comparable levels of IL-8 mRNA. A strong correlation was observed between the elongated particle (aspect ratio ≥3:1) dose metrics of length and external surface area. Expression of the IL-8 data with respect to either of these metrics eliminated the differential response between the RTI amosite sample and the other samples that was observed when HAEC were exposed on an equal mass basis. CONCLUSIONS: On an equal mass basis, LA is as potent as the UICC amosite reference sample at inducing a pro-inflammatory response in HAEC but is less potent than the RTI amosite sample. The results of this study show that the particle length and particle surface area are highly correlated metrics that contribute significantly to the toxicological potential of these amphibole samples with respect to the inflammogenic response induced in airway epithelial cells.

Morphological and chemical mechanisms of elongated mineral particle toxicities.

Much of our understanding regarding the mechanisms for induction of disease following inhalation of respirable elongated mineral particles (REMP) is based on studies involving the biological effects of asbestos fibers. The factors governing the disease potential of an exposure include duration and frequency of exposures; tissue-specific dose over time; impacts on dose persistence from in vivo REMP dissolution, comminution, and clearance; individual susceptibility; and the mineral type and surface characteristics. The mechanisms associated with asbestos particle toxicity involve two facets for each particle's contribution: (1) the physical features of the inhaled REMP, which include width, length, aspect ratio, and effective surface area available for cell contact; and (2) the surface chemical composition and reactivity of the individual fiber/elongated particle. Studies in cell-free systems and with cultured cells suggest an important way in which REMP from asbestos damage cellular molecules or influence cellular processes. This may involve an unfortunate combination of the ability of REMP to chemically generate potentially damaging reactive oxygen species, through surface iron, and the interaction of the unique surfaces with cell membranes to trigger membrane receptor activation. Together these events appear to lead to a cascade of cellular events, including the production of damaging reactive nitrogen species, which may contribute to the disease process. Thus, there is a need to be more cognizant of the potential impact that the total surface area of REMP contributes to the generation of events resulting in pathological changes in biological systems. The information presented has applicability to inhaled dusts, in general, and specifically to respirable elongated mineral particles.

Direct application of biota-sediment accumulation factors.

In the early stages of risk assessments for sites with contaminated sediments, predictions of risks are often complicated or limited by sparse or inadequate bioaccumulation data. These limitations often require risk assessors to estimate bioaccumulation relationships in order to complete the assessments of risk. In the present study, the errors are evaluated with the simple (direct) application of field measured biota-sediment accumulation factors (BSAFs) to other species at a specific location, and to the same species and/or other species at other locations within a site and to other sites. The median (90th percentile) differences in directly applying BSAFs to other species at a specific location were < or =2.1x (< or =5.1x) for fish and mussel species groups. The median (90th percentile) differences for applications across locations within a site for a specific species and to other species were < or =3.3x (< or =10x) for fish, mussel, and decapod crustacean groups. For direct application across sites, slightly larger median (90th percentile) differences were observed, i.e., < or =4.0x (< or =12x). The analysis was performed using a data set of 17,848 BSAFs spanning 392 chemicals/chemical combinations and 71 species.

Organic carbon-water concentration quotients (II(SOC)S and pi(poc)S): measuring apparent chemical disequilibria and exploring the impact of black carbon in Lake Michigan.

Chemical concentration quotients measured between water and total organic carbon (TOC) in sediment (II(SOC)) or suspended particulates (pi(poc)) in southern Lake Michigan reveal up to 2 orders of magnitude differences for polychlorinated biphenyl (PCB), dibenzo-p-dioxin (PCDD), dibenzofuran (PCDF), and polycyclic aromatic hydrocarbon (PAH) compounds with similar octanol-water partition coefficients (K(ow)S). Apparent disequilibria for PAHs, PCDDs, and PCDFs, determined as measured II(SOC)S or pi(poc)S divided by their organic carbon equilibrium partitioning values, are significantly greater than disequilibria of PCBs with similar K(ow)S. Apparent disequilibria, when adjusted for black carbon content by using published black carbon nonlinear partition coefficients (K(f,bc)S) and a Freundlich exponent (n(f)) value = 0.7, still exceed equilibrium predictions for the PAHs, PCBs, and PCDDs but with the PCDF disequilibria uniquely below equilibrium. While Monte Carlo analysis of all the variables associated with the black carbon adjusted disequilibria provides wide confidence intervals for individual chemicals, the large class disequilibria differences between PAHs and PCDFs with respect to the PCBs and PCDDs are highly significant. Use of the PCDD K(f,bc)S for calculating both the PCDF and PCDD disequilibria eliminates their extreme divergence. On the basis of the complexity of carbonaceous geosorbent effects and the apparent variable degrees of chemical sequestration in particles, the disequilibria can be adjusted by chemical class to meet expected near equilibrium conditions between suspended particles and water in the hypolimnion. Although these adjustments to the disequilibria calculations produce consistent and plausible values, the complexities of variable carbonaceous geosorbent affinities for these chemicals in Lake Michigan presently favor use of measured, rather than a priori modeled, steady-state total organic carbon-water concentration quotients indexed to TOC as biogenic organic carbon.

Tissue distribution and metabolism of benzo[a]pyrene in embryonic and larval medaka (Oryzias latipes).

The need to understand chemical uptake, distribution, and metabolism in embryonic and larval fish derives from the fact that these early life stages often exhibit greater sensitivity to xenobiotic compounds than do adult animals. In this study, a 6-h acute waterborne exposure immediately after fertilization was used to quickly load the egg with benzo[a]pyrene (BaP). This exposure was used to mimic the initial egg concentration of a persistent bioaccumulative toxicant that could result from maternal transfer. We used multiphoton laser scanning microscopy (MPLSM) in combination with conventional analytical chemistry methods to characterize the tissue distribution of BaP and its principal metabolites in medaka embryos and post-hatch larvae. Embryonic metabolism of BaP was evident by MPLSM prior to liver formation or heart development. A major product of this metabolism was identified by liquid chromatography/mass spectrometry as BaP-3-glucuronide. MPLSM showed that metabolites were sequestered within the yolk, biliary system, and gastrointestinal tract. When the gastrointestinal tract became patent a few days after hatch, the metabolites were rapidly eliminated. These findings indicate that some of the earliest embryonic tissues are metabolically competent and that redistribution of BaP and its metabolic products occurs throughout development. Rapid metabolism of BaP substantially reduces the body burden of parent chemical in the developing embryo, potentially reducing toxicity. It remains unclear whether metabolism of BaP in medaka embryos leads to the formation of DNA adducts associated with genotoxic effects or yields metabolites that later lead to other toxicity in juveniles or adults.

A hybrid empirical-mechanistic modeling approach for extrapolating biota-sediment accumulation factors and bioaccumulation factors across species, time, and/or ecosystems.

An approach is presented for extrapolating field-measured biota-sediment accumulation factors (BSAFs) and bioaccumulation factors (BAFs) across species, time, and/or ecosystems. This approach, called the hybrid bioaccumulation modeling approach, uses mechanistic bioaccumulation models to extrapolate field-measured bioaccumulation data (i.e., BSAFs and BAFs) to new sets of ecological conditions. The hybrid approach predicts relative differences in bioaccumulation using food web models with two sets of ecological conditions and parameters: One set for the ecosystem where the BSAFs and/or BAFs were measured, and the other set for the ecological conditions and parameters for which the extrapolated BSAFs and/or BAFs are desired. The field-measured BSAF (or BAF) is extrapolated by adjusting the measured BSAF (or BAF) by the predicted relative difference, which is derived from two separate solutions of the food web model. Extrapolations of polychlorinated biphenyl BSAFs and BAFs for lake trout (Salvelinus namaycush) from southern Lake Michigan to Green Bay of Lake Michigan (Green Bay, WI, USA) walleye (Stizostedion vitreum) and brown trout (Salmo trutta), as well as Hudson River largemouth bass (Micropterus salmoides) and yellow perch (Perca flavescens), resulted in generally better agreement between measured and predicted BSAFs and BAFs with the hybrid approach.

Comparison of biota-sediment accumulation factors across ecosystems.

Sets of biota-sediment accumulation factors (BSAFs) for fish were compared across ecosystems for nonionic organic chemicals. The sets of BSAFs, when plotted against each other, in log-log space, formed linear relationships and demonstrated that the relative scaling or ranking of the individual BSAFs within a set are consistent, if not the same, across ecosystems. This behavior holds for chemicals that either are, or are not, metabolized by fish. These results demonstrate that sets of BSAF values can differ but with parallel shifts in magnitude between ecosystems (for example, all of the BSAFs in the set are uniformly larger in one ecosystem, while in another they all are uniformly smaller) in response to underlying differences in ecosystem conditions and parameters such as trophic level, diet of the organisms, and distribution of the chemical between the sediment and water column.

Biota--sediment accumulation factors for polychlorinated biphenyls, dibenzo-p-dioxins, and dibenzofurans in southern Lake Michigan lake trout (Salvelinus namaycush).

A set of high-quality, age-specific biota-sediment accumulation factors (BSAFs) for polychlorinated biphenyls (PCBs), dibenzo-p-dioxins (PCDDs), and dibenzofurans (PCDFs) have been determined from concentrations measured with high-resolution gas chromatography/high-resolution mass spectrometry, by use of the 13C isotopic dilution technique, in lake trout and surficial (0-2 cm) sediment samples from southern Lake Michigan. BSAFs ranged from <0.1 to 18 for PCBs and from <0.001 to 0.32 for PCDDs and PCDFs detected in the fish. PCBs with zero or one chlorine in an ortho position had smaller BSAFs than other PCBs. PCDDs and PCDFs with chlorines at the 2,3,7,8-positions had larger BSAFs than most other PCDDs and PCDFs. The fidelity of the relative bioaccumulation potential data between independent lake trout samples, within and among age classes, suggests that differences in slight rates of net metabolism in the food chain are important and contribute to the apparent differences in BSAFs, not only for PCDDs and PCDFs but also possibly for some PCBs. A complicating factor for non-ortho- and mono-ortho-PCBs is the uncertain contribution of enhanced affinity for black carbon (and possibly volatility) acting in concert with metabolism to reduce measured BSAFs for lake trout. On the basis of the association between chemicals with apparent slight rates of metabolism and measured dioxin-like toxicity, several PCDFs with similar measured BSAFs but unknown toxicity may be candidates for toxicity testing.

Use of multi-photon laser-scanning microscopy to describe the distribution of xenobiotic chemicals in fish early life stages.

To better understand the mechanisms by which persistent bioaccumulative toxicants (PBTs) produce toxicity during fish early life stages (ELS), dose-response relationships need to be understood in relation to the dynamic distribution of chemicals in sensitive tissues. In this study, a multi-photon laser scanning microscope (MPLSM) was used to determine the multi-photon excitation spectra of several polyaromatic hydrocarbons (PAHs) and to describe chemical distribution among tissues during fish ELS. The multi-photon excitation spectra revealed intense fluorescent signal from the model fluorophore, pentamethyl-difluoro-boro-indacene (BODIPY), less signal from benzo[a]pyrene and fluoranthene, and no detectable signal from pyrene. The imaging method was tested by exposing newly fertilized medaka (Oryzias latipes) eggs to BODIPY or fluoranthene for 6 h, followed by transfer to clean media. Embryos and larvae were then imaged through 5 days post-hatch. The two test chemicals partitioned similarly throughout development and differences in fluorescence intensity among tissues were evident to a depth of several hundred microns. Initially, the most intense signal was observed in the oil droplet within the yolk, while a moderate signal was seen in the portion of the yolk containing the yolk-platelets. As embryonic development progressed, the liver biliary system, gall bladder, and intestinal tract accumulated strong fluorescent signal. After hatch, once the gastrointestinal tract was completely developed, most of the fluorescent signal was cleared. The MPLSM is a useful tool to describe the tissue distribution of fluorescent PBTs during fish ELS.

The relationship of bioaccumulative chemicals in water and sediment to residues in fish: a visualization approach.

A visualization approach is developed and presented for depicting and interpreting bioaccumulation relationships and data (i.e., bioaccumulation factors [BAFs], biota-sediment accumulation factors [BSAFs], and chemical residues in fish) using water-sediment chemical concentration XY plots. The approach is based on five basic parameters that affect bioaccumulation of nonionic organic chemicals: The distribution of chemical between sediment and water, the hydrophobicity of the compound (expressed as the n-octanol/water partition coefficient, K(OW)), the relationship of food chains to water and sediment, the length of the food chains, and the degree to which the chemical is metabolized. The visualization approach using water-sediment XY plots captures and visually presents the existing bioaccumulation knowledge in a form that is readily understandable from chemical, biological, and ecological aspects and, therefore, useful in the assessment, communication, and management of risk for persistent bioaccumulative toxicants.

Evaluation of two methods for prediction of bioaccumulation factors.

Two methods for deriving bioaccumulation factors (BAFs) used by the U.S. Environmental Protection Agency in development of water quality criteria were evaluated using polychlorinated biphenyl (PCB) data from the Hudson River and Green Bay ecosystems. One method predicts BAF(L)fd values (BAFs based upon concentrations of freely dissolved chemical in ambient water and in the lipid fraction of tissue) using field-measured biota-sediment accumulation factors (BSAFs): greater than 90% of the predicted BAF(L)fd values were within a factor of 5 of their measured values for both ecosystems. The second method predicts BAF(L)fd values as the chemical's 1-octanol/water partition coefficient (Kow) times a food chain multiplier: greater than 90% of the predicted BAF(L)fd values were within a factor of 5 of their measured values for the Green Bay ecosystem and for three of the six sampling locations on the Hudson River. Poorer predictive ability with the Kow method for the other three sampling locations was believed to be due to the existence of environmental conditions not representative of the longer term ecosystem conditions assumed for the method. BAF(L)fd and BAF(T)t values (BAFs based upon concentrations of total chemical in ambient water and in wet tissue) were compared. The within ecosystem and across ecosystems comparisons demonstrated a 2-5-fold decrease in variability (expressed as ratios of coefficients of variation, percentile ranges, and confidence ranges) for predicted BAF(L)fd values in comparison to BAF(T)t values.

Effects of aryl hydrocarbon receptor-mediated early life stage toxicity on lake trout populations in Lake Ontario during the 20th century.

Lake trout embryos and sac fry are very sensitive to toxicity associated with maternal exposures to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and structurally related chemicals that act through a common aryl hydrocarbon receptor (AHR)-mediated mechanism of action. The loading of large amounts of these chemicals into Lake Ontario during the middle of the 20th century coincided with a population decline that culminated in extirpation of this species around 1960. Prediction of past TCDD toxicity equivalence concentrations in lake trout eggs (TEC(egg)s) relative to recent conditions required fine resolution of radionuclide-dated contaminant profiles in two sediment cores; reference core specific biota--sediment accumulation factors (BSAFs) for TCDD-like chemicals in lake trout eggs; adjustment of the BSAFs for the effect of temporal changes in the chemical distributions between water and sediments; and toxicity equivalence factors based on trout early life stage mortality. When compared to the dose-response relationship for overt early life stage toxicity of TCDD to lake trout, the resulting TEC(egg)s predict an extended period during which lake trout sac fry survival was negligible. By 1940, following more than a decade of population decline attributable to reduced fry stocking and loss of adult lake trout to commercial fishing, the predicted sac fry mortality due to AHR-mediated toxicity alone explains the subsequent loss of the species. Reduced fry survival, associated with lethal and sublethal adverse effects and possibly complicated by other environmental factors, occurred after 1980 and contributed to a lack of reproductive success of stocked trout despite gradually declining TEC(egg)s. Present exposures are close to the most probable no observable adverse effect level (NOAEL TECegg = 5 pg TCDD toxicity equivalence/g egg). The toxicity predictions are very consistent with the available historical data for lake trout population levels in Lake Ontario, stocking programs, and evidence for recent improvement in natural reproduction concomitant with declining levels of persistent bioaccumulative chemicals in sediments and biota.