In vitro prediction of polycyclic aromatic hydrocarbon bioavailability of 14 different incidentally ingested soils in juvenile swine.

Predicting mammalian bioavailability of PAH mixtures from in vitro bioaccessibility results has proven to be an elusive goal. In an attempt to improve in vitro predictions of PAH soil bioavailability we investigated how energetic input influences PAH bioaccessibility by using a high and low energetic shaking method. Co-inertia analysis (COIA), and Structural Equation Modeling (SEM) were also used to examine PAH-PAH interactions during ingestion. PAH bioaccessibility was determined from 14 historically contaminated soils using the fed organic estimation of the human simulation test (FOREhST) with inclusion of a silicone rod as a sorption sink and compared to bioavailability estimates from the juvenile swine model. Shaking method significantly affected PAH bioaccessibility in the FOREhST model, with PAH desorption from the high energy FOREhST almost an order of magnitude greater compared to the low energy FOREhST. PAH-PAH interactions significantly influenced PAH bioavailability and when these interactions were used in a linear model, the model predicted benzo(a)anthracene bioavailability with an slope of 1 and r2 of 0.66 and for benzo(a)pyrene bioavailability has a slope of 1 and r2 of 0.65. Lastly, to confirm the effects as determined by COIA and SEM, we spiked low levels of benzo(a)anthracene into historically contaminated soils, and observed a significant increase in benzo(a)pyrene bioaccessibility. By accounting for PAH interactions, and reducing the energetics of in vitro extractions, we were able to use bioaccessibility to predict bioavailability across 14 historically contaminated soils. Our work suggests that future work on PAH bioavailability and bioaccessibility should focus on the dynamics of how the matrix of PAHs present in the soil interact with mammalian systems. Such interactions should not only include the chemical interactions discussed here but also the interactions of PAH mixtures with mammalian uptake systems.

Do biomarkers of exposure and effect correlate with internal exposure to PAHs in swine?

Humans are commonly exposed to polycyclic aromatic hydrocarbons (PAHs), a family of compounds present as mixtures in the environment. This study exposed swine to PAH mixtures in single and subacute dose regimens and collected liver and ileum tissue to measure cytochrome P450 mRNA expression and enzyme activity as biomarkers of exposure and DNA adducts and oxidized proteins as biomarkers of effect. Micronucleated reticulocytes were measured as systemic biomarkers of effect. Duration of exposure did not influence biomarkers of exposure, though exposure duration produced significant increases in DNA adducts and oxidative stress. Micronucleated reticulocyte numbers were not affected by exposure length.

Is received dose from ingested soil independent of soil PAH concentrations?-Animal model results.

Polycyclic aromatic hydrocarbon (PAH) bioavailability from ingested soils will vary between soils; however, the nature of this variation is not well characterized. A juvenile swine model was used to link external exposure to internal benzo[a]pyrene (BaP) and anthracene exposure following oral PAH ingestion of 27 different impacted site soils, soots, or spiked artificial soils. Internal exposure of BaP and anthracene, represented by area under the plasma-time curve, did not relate to soil concentration in impacted site soils, but did relate in spiked artificial soil. Point of departure modeling identified soil PAH concentrations greater than 1900 mg kg(-1) as the point where area under the curve becomes proportional to external dose. A BaP internal exposure below 1900 mg kg(-1) had an upper 95% confidence interval estimate of 33% of external exposure. Weak relationships between soil:simulated gastrointestinal fluid PAH partitioning and area under the curve values suggest that differences in internal PAH exposure between soils may not be dominated by differences in PAH partitioning. The data seem to best support exposure assessment assuming constant internal PAH exposure below soil concentrations of 1900 mg kg(-1) . However, because constant internal exposure would challenge several existing paradigms, a bioavailability estimate of 33% of the external exposure is suggested as a likely workable solution. Environ Toxicol Chem 2016;35:2261-2269. © 2016 SETAC.

Predicting Polycyclic Aromatic Hydrocarbon Bioavailability to Mammals from Incidentally Ingested Soils Using Partitioning and Fugacity.

Soil and dust ingestion is one of the major human exposure pathways to contaminated soil; however, pollutant transfer from ingested substances to humans cannot currently be confidently predicted. Soil polycyclic aromatic hydrocarbon (PAH) bioavailability is likely dependent upon properties linked to chemical potential and partitioning such as fugacity, fugacity capacity, soil organic carbon, and partitioning to simulated intestinal fluids. We estimated the oral PAH bioavailability of 19 historically contaminated soils fed to juvenile swine. Between soils, PAH blood content, with the exception of benzo(a)pyrene, was not linked to fugacity. In contrast, between individual PAHs, using partitioning explained PAH blood content (area under the curve = 0.47 log fugacity + 0.34, r(2) = 0.68, p < 0.005, n = 14). Soil fugacity capacity predicts PAH soil concentration with an average slope of 0.30 (µg PAH g(-1) soil) Pa(-1) and r(2)'s of 0.61-0.73. Because PAH blood content was independent of soil concentration, soil fugacity correlated to PAH bioavailability via soil fugacity's link to soil concentration. In conclusion, we can use fugacity to explain PAH uptake from a soil into blood. However, something other than partitioning is critical to explain the differences in PAH uptake into blood between soils.

Using near infrared transmittance to generate sorted fractions of Fusarium-infected wheat and the impact on broiler performance.

The objective of this study was to investigate the effects of naturally contaminated Fusarium wheat containing deoxynivalenol (DON) on growth and performance of broiler chickens from 0 to 35 d. The BoMill TriQ individual kernel sorting technology uses near infrared transmittance (NIT) spectra to separate Fusarium-damaged kernels (FDK) from healthy kernels based on individual kernel CP. Three Fusarium-contaminated wheat sources were individually sorted into 3 test fractions: outlier (10% of the source), high mycotoxin (20% of the source), and low mycotoxin (70% of the source). These fractions were reconstituted into 4 ratios-M0, M20, M40, and M60-relating to the proportion of the high mycotoxin fraction in the reconstituted diets. These 12 reconstituted wheat sources with varying levels of DON were incorporated at ∼70% (starter) or ∼75% (grower/finisher) into diets. The fractions of wheat used had FDK ranging from 0.1 to 25.8% and DON from 0.0 to 14.3 ppm. A total of 480 newly hatched Ross 308 male broilers were randomly divided into 96 cages. Each test diet was assigned to 8 replicates with 5 birds per replicate cage. At 21 d, 180 birds were transferred to 36 cages, allowing 3 replicates of 5 birds per diet until 35 d. A factorial arrangement analysis compared the 3 wheat sources and 4 ratios produced from each sorted wheat. Growth and performance were evaluated as BW (g), feed intake (FI; g/bird/day), feed conversion ratio (FCR; g:g), AME (kcal ME/kg diet), nitrogen retention (NR; %), and mortality (%) for 0 to 21 d and 21 to 35 d. Results indicate no significant difference in BW, FI, and FCR (P > 0.05). Significant differences were seen in AME and NR (P < 0.01). This study demonstrates the potential of this novel sorting technology to produce naturally contaminated diets with a large range of mycotoxin concentrations from a single wheat source to enable future investigations of mycotoxin exposure in any species.

The use of near infrared transmittance kernel sorting technology to salvage high quality grain from grain downgraded due to Fusarium damage.

The mycotoxins associated with specific Fusarium fungal infections of grains are a threat to global food and feed security. These fungal infestations are referred to as Fusarium Head Blight (FHB) and lead to Fusarium Damaged Kernels (FDK). Incidence of FDK >0.25% will lower the grade, with a tolerance of 5% FDK for export feed grain. During infestation, the fungi can produce a variety of mycotoxins, the most common being deoxynivalenol (DON). Fusarium Damaged Kernels have been associated with reduced crude protein (CP), lowering nutritional, functional and grade value. New technology has been developed using Near Infrared Transmittance (NIT) spectra that estimate CP of individual kernels of wheat, barley and durum. Our objective is to evaluate the technology's capability to reduce FDK and DON of downgraded wheat and ability to salvage high quality safe kernels. In five FDK downgraded sources of wheat, the lowest 20% CP kernels had significantly increased FDK and DON with the high CP fractions having decreased FDK and DON, thousand kernel weights (TKW) and bushel weight (Bu). Strong positive correlations were observed between FDK and DON (r = 0.90); FDK and grade (r = 0.62) and DON and grade (r = 0.62). Negative correlations were observed between FDK and DON with CP (r = -0.27 and -0.32); TKW (r = -0.45 and -0.54) and Bu (r = -0.79 and -0.74). Results show improved quality and value of Fusarium downgraded grain using this technology.

The bioavailability of polycyclic aromatic hydrocarbons from different dose media after single and sub-chronic exposure in juvenile swine.

Humans are constantly exposed to contaminants in the environment, which may lead to changes in physiological processes by altering enzyme activities that could affect bioavailability. However, bioavailability estimates are typically made from a single exposure to an animal model, which may lead to overestimating bioavailability. This study uses juvenile swine to model human exposure to benzo[a]pyrene (BaP) and anthracene in certified reference material (CRM), spiked soil, spiked food, or spiked corn oil after one and seven days of dosing. Area under the curve (AUC) was calculated after one and seven days of exposure for both BaP and anthracene for each exposure media. Whereas there were significant differences in AUC between different media, there were no significant changes in AUC after sub-chronic exposure to BaP or anthracene. Average BaP bioavailability for CRM, spiked soil, spiked food and corn oil was 71%, 0.72%, 0.03% and 0.97% respectively. Average anthracene bioavailability was 1.7% and 43% for corn oil and CRM respectively. Anthracene was not detected above background in swine exposed to spiked food and spiked soil. Thus, this study indicates that exposure media impacts bioavailability, but there is no statistical evidence that sub-chronic exposure affects systemic exposure.

An investigation of the effect of gastrointestinal microbial activity on oral arsenic bioavailability.

In vitro gastrointestinal (GI) microbial activity in the colon compartment facilitates the arsenic release from soils into simulated GI fluids. Consequentially, it is possible that in vitro models that neglect to include microbial activity underestimate arsenic bioaccessibility when calculating oral exposure. However, the toxicological relevance of increased arsenic release due to microbial activity is contingent upon the subsequent absorption of arsenic solubilized in the GI lumen. The objectives of this research are to: (1) assess whether microbes in the in vitro small intestine affect arsenic solubilization from soils, (2) determine whether differences in the GI microbial community result in differences in the oral bioavailability of soil-borne arsenic. In vitro GI microbial activity in the distal small intestine increased arsenic release from soils; however, these effects were unlikely to be relevant since they were transient and demonstrated small effect sizes. In vivo arsenic absorption for juvenile swine was unaffected by antibiotic treatment. Therefore, it appears that microbial effects on arsenic release do not result in increased arsenic bioavailability. However, it remains to be seen whether the results for the limited set of soils described herein can be extrapolated to arsenic contaminated sites in general.

Combined effect of short-term dehydration and sublethal acute oral dicrotophos exposure confounds the diagnosis of anticholinesterase exposure in common quail (Coturnix coturnix) using plasma cholinesterase activity.

Common Quail (Coturnix coturnix) were subjected to controlled and replicated experiments in the summer of 2008 to investigate the effects of short-term dehydration on cholinesterase activity in brain and plasma and the interaction between dehydration and exposure to the organophosphorus pesticide dicrotophos in these same tissues. Our objective was to determine if dehydration could confound the diagnosis of anticholinesterase exposure using inhibition of cholinesterase activity in quail tissues. The effect of dehydration was quantified using measures of plasma osmolality and hematocrit. Dicrotophos exposure caused significant inhibition of cholinesterase activity in brain, while the effects of dehydration and interaction were not significant. Dehydration caused significant duration-dependent increases in plasma osmolality and hematocrit. Dehydration also caused a significant increase in plasma cholinesterase activity. Variation in the change in plasma cholinesterase activity in response to dehydration was significantly and positively correlated with dehydration-induced variation in both the change in plasma osmolality and the change in hematocrit. These correlations suggest that plasma cholinesterase activity in quail is not limited to plasma but occupies some larger pool of the extracellular fluid volume, and we suggest lymph is part of that pool. The effects of dehydration on plasma cholinesterase activity masked the inhibitory effects of dicrotophos. Here, the combination of dehydration and dicrotophos exposure produced plasma cholinesterase activity that was not significantly different from reference and pre-exposure values, confounding the diagnosis of anticholinesterase exposure in dehydrated, dicrotophos-exposed quail. A method to adjust plasma cholinesterase activities for the confounding effects of dehydration and enable the diagnosis of anticholinesterase exposure in dehydrated, dicrotophos-exposed quail was developed. Clinicians and practitioners responsible for the diagnosis of anticholinesterase exposure in birds are cautioned that dehydration, commonly observed in sick wildlife, may mask the effect of anticholinesterases on plasma cholinesterase activity.

Human exposure assessment: a case study of 8 PAH contaminated soils using in vitro digestors and the juvenile swine model.

In vitro digestors can be used to provide bioaccessibility values to help assess the risk from incidental human ingestion of contaminated soils. It has been suggested that these digestors may need to include a lipid sink to mimic human uptake processes. We compare the correspondence between in vivo polycyclic aromatic hydrocarbon (PAH) uptake for eight different PAH contaminated soils with PAH release in in vitro digestors in the presence and absence of a lipid sink. Lipid sinks were essential to the success of the in vitro digestors in predicting juvenile swine PAH uptake. In the presence of the lipid sink, results of the In Vitro Digestion model (IVD) closely corresponded with a slope of 0.85 (r(2) = 0.45, P < 0.07) to the in vivo results. The Relative Bioaccessibility Leaching Procedure (RBALP) results did not correspond to the in vivo study but did tightly reflect total soil PAH concentration. We conclude that the basis of this difference between digestors is that the RBALP used an aggressive extraction technique that maximized PAH release from soil. Systemic uptake in juvenile swine was not linked to soil PAH concentration but rather to the thermodynamic properties of the soil.

Toxicity and immune system effects of dietary deltamethrin exposure in tiger salamanders (Ambystoma tigrinum).

One theory proposed to explain the global declines in amphibian populations involves contaminant-induced immune alteration and subsequent increased susceptibility to infectious disease. The goal of this study was twofold, to (1) study acute oral toxicity of deltamethrin (cyclopropanecarboxylic acid, 3-(2,2-dibromoethenyl)-2,2-dimethyl cyano(3-phenoxyphenyl)methyl ester) in tiger salamanders (Ambystoma tigrinum), and (2) evaluate whether the insecticide deltamethrin produces immunosuppression in these animals. In the acute toxicity study, tiger salamanders receiving single doses of deltamethrin ranging from 1 to 35 mg/kg displayed intention tremors, hypersalivation, ataxia, choreoathetosis (writhing), severe depression (immobility with minimal response to stimuli), and death. For acute effects, based on clinical signs, the median lethal dose (LD(50)) and lowest observed adverse effect level (LOAEL) were estimated to be 5 to 10 mg/kg and 1 mg/kg, respectively. The LOAEL in animals dosed 3 times per week for 4 wk was 400 microg/kg/d. The endpoints for the immunotoxicity study included lymphoid organ mass and histopathology, hematological variables, and functional assays of phagocytosis, oxidative burst, and lymphoblastic transformation. Tiger salamanders in 4 treatment groups (0, 4, 40, or 400 microg/kg/d) were dosed with deltamethrin via the diet 3 times per week for 4 wk. Deltamethrin exposure resulted in increased liver mass, packed cell volume, and total plasma protein concentration, but these effects were not dose dependent. The relative mass of kidney and spleen, plasma albumin and globulin concentrations, and circulating leukocyte numbers were not affected by deltamethrin exposure, nor were phagocytosis, oxidative burst, and lymphoblastic transformation. This study shows that at moderate levels of exposure, deltamethrin may be neurotoxic to tiger salamanders. However, based on the immune assays considered in this study there was no evidence of immunosuppression from dietary exposure to environmentally relevant concentrations of deltamethrin. In light of these findings, it is unlikely that exposure to environmental concentrations of deltamethrin has produced immunosuppression and contributed to the emergence of iridovirus outbreaks in tiger salamander populations.

Associations between immune function in yearling beef cattle and airborne polycyclic aromatic hydrocarbons and PM1.0 near oil and natural gas field facilities.

Researchers determined the potential associations between exposure to airborne fine particulate matter (ie, particulate matter that is < 1 microm in diameter, or PM1.0) and polycyclic aromatic hydrocarbons (PAHs) and immune system function in beef cattle by using blood samples collected from yearling cattle in 22 herds in the spring of 2002. The herds were located at variable distances from industry field facilities in the major oil- and gas-producing areas of western Canada. The researchers evaluated immune system competence by measuring populations of B-lymphocytes and T-lymphocyte subtypes (CD4, CD8, gammadelta, and WC1) in peripheral circulation (n = 469), and systemic antibody production in response to vaccine administration (n = 446). They used particulate air monitors to estimate the exposure of the cattle to airborne contaminants by determining mean monthly concentrations of PM1.0 and 24 different PAHs from January to June. The mean concentration of PAHs measured in the ambient air of herds monitored in this study was low, with naphthalene being present in the highest concentration (geometric mean = 5.6 ng/m3; geometric standard deviation = 38), followed by 1-methylnaphthalene (geometric mean = 2.2 ng/m3; geometric standard deviation = 12). The geometric mean monthly exposure to PM1.0 was 7.1 microg/m3 (geometric standard deviation = 1.5) for the same period. The researchers detected no significant plausible associations between exposure to any measured airborne contaminants and immune system function.

Associations between in utero exposure to airborne emissions from oil and gas production and processing facilities and immune system outcomes in neonatal beef calves.

To determine if oil- and gas-facility emissions prospectively measured as airborne sulfur dioxide and volatile organic compounds (VOCs) were associated with immunomodulation in neonatal beef calves, veterinarians collected samples from 325 calves from 60 western Canadian herds in the spring of 2002. Researchers assessed immune system effects by enumerating B-lymphocytes and specific T-lymphocyte subtypes (CD4, CD8, gammadelta, and WC1) in peripheral circulation using flow cytometry. They estimated in utero exposure for each calf from the earliest possible breeding date of the dam to the calving date, using air-quality data from passive monitors installed in pastures and wintering areas. Numbers of circulating CD4 and CD8 T-lymphocytes were 42% and 43% lower, respectively, in calves exposed to the highest quartile (> or = 0.378 microg/m3) of VOCs measured as airborne concentrations of benzene compared with calves exposed to concentrations in the lowest quartile (< 0.276 microg/m3). Similarly, the number of CD4 T-lymphocytes was 40% lower in calves exposed to VOCs measured as concentrations of toluene in the highest quartile (> or = 0.713 microg/m3), compared with calves exposed to concentrations in the lowest quartile (< 0.348 microg/m3). There was no statistically significant association between in utero exposure and numbers of B-lymphocytes and gammadelta and WC1 T-lymphocyte subtypes in calf blood samples.

Associations between immune function in yearling beef cattle and airborne emissions of sulfur dioxide, hydrogen sulfide, and VOCs from oil and natural gas facilities.

Researchers assessed the associations between airborne emissions from oil and gas field facilities and the structure and function of the immune system of yearling beef cattle in 27 herds during spring 2002. They evaluated the immune systems of these animals by enumerating B lymphocytes and T-lymphocyte subtypes (CD4, CD8, gammadelta, and WC1) in peripheral circulation and by measuring systemic antibody production in response to vaccination. Researchers prospectively measured exposure to sulfur dioxide, hydrogen sulfide, and volatile organic compounds (VOCs) by using air-quality data from passive monitors installed in pastures and wintering areas. They estimated the mean exposure of each animal over the 6-month period before the start of sample collection. The researchers used mixed models, which adjusted for clustering by herd and accounted for known risk factors, to examine potential associations between exposure to airborne sulfur dioxide, VOCs (measured as concentrations of benzene and toluene) and hydrogen sulfide, as well as proximity to emission sources (well-site density), and the immune system outcomes. Increasing exposure to VOCs measured as toluene was associated with significant CD4 T lymphocytopenia. The number of CD4 T lymphocytes was 30% lower in cattle exposed to VOCs measured as toluene in the highest quartile (> 0.823 microg/m3) than in cattle exposed in the lowest quartile (< 0.406 microg/m3).

Biogeochemical toxicity and phytotoxicity of nitrogenous compounds in a variety of arctic soils.

Ammonium nitrate (NH(4)NO(3)) is a common water pollutant associated with many industrial and municipal activities. One solution to reduce exposure of sensitive aquatic systems to nitrogenous compounds is to atomize (atmospherically disperse in fine particles) contaminated water over the Arctic tundra, which will reduce nitrogen loading to surface water. The toxicity of ammonium nitrate to Arctic soils, however, is poorly understood. In the present study, we characterized the biogeochemical toxicity and phytotoxicity of ammonium nitrate solutions in four different Arctic soils and in a temperate soil. Soil was exposed to a range of ammonium nitrate concentrations over a 90-d period. Dose responses of carbon mineralization, nitrification, and phytotoxicity endpoints were estimated. In addition to direct toxicity, the effect of ammonium nitrate on ecosystem resilience was investigated by dosing nitrogen-impacted soils with boric acid. Ammonium nitrate had no effect on carbon mineralization activity and only affected nitrification in one soil, a polar desert soil from Cornwallis Island, Northwest Territories, Canada. In contrast, ammonium nitrate applications (43 mmol N/L soil water) significantly impaired seedling emergence, root length, and shoot length of northern wheatgrass (Elymus lanceolatus). Concentrations of ammonium nitrate in soil water that inhibited plant parameters by 20% varied between 43 and 280 mmol N/L soil water, which corresponds to 2,100 to 15,801 mg/L of ammonium nitrate in the application water. Arctic soils were more resistant to ammonium nitrate toxicity compared with the temperate soil under these study conditions. It is not clear, however, if this represents a general trend for all polar soils, and because nitrogen is an essential macronutrient, nitrogenous toxicity likely should be considered as a special case for soil toxicity.

Increased sensitivity and variability of phytotoxicity responses in Arctic soils to a reference toxicant, boric acid.

Industrial and human activities in the Arctic regions may pose a risk to terrestrial Arctic ecosystem functions. One of the most common terrestrial toxicological end points, primary productivity, typically is assessed using a plant phytotoxicity test. Because of cryoturbation, a soil mixing process common in polar regions, we hypothesized that phytotoxicity test results in Arctic soils would be highly variable compared to other terrestrial ecosystems. The variability associated with phytotoxicity tests was evaluated using Environment Canada's standardized plant toxicity test in three cryoturbated soils from Canada's Arctic exposed to a reference toxicant, boric acid. Northern wheatgrass (Elymus lanceolatus) not only was more sensitive to toxicants in Arctic soils, its response to toxicants was more variable compared to that in temperate soils. The phytotoxicity of boric acid in cryosols was much greater than commonly reported in other soils, with a boric acid concentration of less than 150 microg/g soil needed to inhibit root and shoot growth by 20%. Large variability also was found in the phytotoxicity test results, with coefficients of variation for 10 samples ranging from 160 to 79%. The increased toxicity of boric acid in cryosols and variability in test response was not explained by soil properties. Based on our admittedly limited data set of three different Arctic soils, we recommend that more than 30 samples be taken from each control and potentially impacted area to accurately assess contaminant effects at sites in northern Canada. Such intensive sampling will insure that false-negative results for toxicant impacts in Arctic soils are minimized.

Two novel biomarkers in developing wood frogs.

Two novel biomarkers (whole-body thyroid hormone and triglyceride concentrations) were developed for use in wood frog tadpoles. The development of these biomarkers was based on previous work done in fish and lizards and incorporated triglyceride and thyroid hormone extraction into 1 homogenization step with 95% ethanol as the solvent. This method is useful for reducing sample-to-sample variability because the same sample can be used for each extraction and assay.

Relative biological effectiveness (RBE) of alpha radiation in cultured porcine aortic endothelial cells.

PURPOSE: Northern peoples can receive elevated radiation doses (1- 10 mSv/y) from transfer of polonium-210 (210Po) through the lichen-caribou-human food chain. Ingested 210Po is primarily blood-borne and thus many of its short range alpha particles irradiate the endothelial cells lining the blood vessels. The relative biological effectiveness (RBE) of alpha particles vs. x-rays was examined in porcine aortic endothelial cells as a surrogate for understanding what might happen to human endothelial cells in northern populations consuming traditional foods. MATERIALS AND METHODS: Cultured porcine aortic endothelial cells were exposed to x-ray and 210Po alpha particle radiation. Alpha irradiation was applied to the cell cultures internally via the culture medium and externally, using thin-bottomed culture dishes. The results given here are based on the external irradiation method, which was found to be more reliable. Dose-response curves were compared for four lethal endpoints (cell viability, live cell fraction, release of lactate dehydrogenase [LDH] and clonogenic survival) to determine the relative biological effectiveness (RBE) of alpha radiation. RESULTS: The alpha RBE for porcine cells varied from 1.6-21, depending on the endpoint: 21.2+/-4.5 for cell viability, 12.9+/-2.7 for decrease in live cell number, 5.3+/-0.4 for LDH release to the medium but only 1.6 +/-0.1 for clonogenic survival. The low RBE of 1.6 was due to x-ray hypersensitivity of endothelial cells at low doses.

Cell death and cytotoxic effects in YAC-1 lymphoma cells following exposure to various forms of mercury.

The effects of 1 min-4 h exposures to four Hg compounds (mercuric chloride [HgCl2], methyl mercuric chloride [CH3HgCl], p-chloromercuribenzoate [p-CMB] and thimerosal [TMS; ethylmercurithiosalicylate]) on cell death, microtubules, actin, CD3 receptor expression, protein tyrosine phosphorylation (PTyr-P) and intracellular calcium ([Ca2+]i) levels were investigated in YAC-1 lymphoma cells using flow cytometry. YOPRO-1 (YP) and propidium iodide (PI) dye uptake indicated all forms of Hg tested were toxic at concentrations ranging from 25.8-48.4 microM, with two distinct patterns of effects. Early apoptosis was prolonged for CH3HgCl- and TMS-treated cells, with more than 50% remaining YP+/PI- after 4h. Both CH3HgCl and TMS induced complete loss of beta-tubulin fluorescence, indicative of microtubule depolymerization and inhibition of tubulin synthesis and/or beta-tubulin degradation, while F-actin fluorescence diminished to a lesser degree and only after loss beta-tubulin. CH3HgCl and TMS induced an almost immediate two-fold increase in CD3 fluorescence, with levels returning to baseline within minutes. With continued exposure, CD3 fluorescence was reduced to approximately 50% of baseline values. Both compounds also increased PTyr-P two- to three-fold immediately, with levels returning to baseline at 4h. Similarly, two- to three-fold increases in [Ca2+]i were noted after 1 min exposure. [Ca2+]i increased progressively, reaching levels five- to eight-fold greater than control values. In contrast, dye uptake was delayed with HgCl2 and p-CMB, although cell death proceeded rapidly, with almost all non-viable cells being late apoptotic (YP+/PI+) by 4h. p-CMB produced early reductions in F-actin, and after 4h, complete loss of F-actin with only partial reduction of total beta-tubulin was seen with both p-CMB and HgCl2. HgCl2 reduced CD3 expression and PTyr-P slightly within minutes, while p-CMB produced similar effects on CD3 only at 4h, at which time PTyr-P was increased two- to three-fold. Both compounds increased [Ca2+]i within minutes, though levels remained under twice the baseline concentration after 15 min exposure. With continued exposure, [Ca2+]i increased to levels two- to five-fold greater than control values. These findings indicate the two groups of Hg compounds may induce cell death by distinct pathways, reflecting interactions with different cellular targets leading to cell death.

Toxicity of sodium chloride to house sparrows (Passer domesticus).

Sodium chloride (NaCl) is widely used as a deicing agent on roadways. There are numerous anecdotal reports of poisoning of passerine birds by road salt in the United States and Canada, but little is known about the toxicity of NaCl to songbirds. The objectives of this study were to determine the lethal dose range for NaCl in a representative passerine species (house sparrow [Passer domesticus]); to determine the clinical, physiologic, and pathologic effects of sublethal and lethal oral NaCl exposure; and to assess the potential for recovery after exposure to granular salt or highly concentrated salt solutions. The up-and-down method was used in a pilot study to estimate the lethal oral dose of granular NaCl in wild caught house sparrows. The toxicity of highly concentrated NaCl solution also was investigated. This was followed by an acute dose response study in which house sparrows were dosed orally with granular NaCl at 0, 500, 1,500, 2,500, or 3,500 mg/kg. Sparrows were deprived of water for 6 hr postexposure (PE) in an attempt to mimic specific winter conditions. Groups of three birds at each dose were euthanized at 1, 3, 6, and 12 hr PE, and samples were collected for histopathology and brain and plasma electrolyte analyses. Results indicated an approximate mean lethal dose (LD50) of 3,000-3,500 mg/kg in water-deprived birds, which is similar to mammalian values. House sparrows dosed with a concentrated solution of NaCl generally died at doses of 8,000 mg/kg. Clinical signs observed at >or=1,500 mg/kg included rapid onset (<30 min) of depression (indicated by reduced activity and reduced response to visual and auditory stimuli), ataxia, inability to fly or perch, and death in as little as 45 min. Birds that survived for 6 hr usually recovered. Plasma Na concentrations >200 mmol/l were consistently associated with clinical signs. Pathologic lesions consisted of edema and distension of the caudoventral thin muscled region of the gizzard and were observed 1 hr PE in most birds dosed with >or=500 mg/kg. Brain Na concentrations in clinically ill sparrows and those that died of NaCl toxicity ranged from 1,297 to 1,615 (mean=1,450; SD=115) ppm wet weight or 5,603 to 6,958 (mean=6,367; SD=454) ppm dry weight, which differed significantly from control birds. No histologic lesions were observed in brain sections of exposed birds, likely reflecting the acute nature of the exposure. However, fluid accumulation beneath the koilin layer of the gizzard was observed in the majority of birds at high dosage levels. These results indicate that passerines ingesting relatively small numbers of road salt granules or small quantities of highly concentrated NaCl solutions are at risk of sodium poisoning.