Robert Budinsky - Academia.edu (original) (raw)

Papers by Robert Budinsky

Research paper thumbnail of Quantitative analysis of unconjugated and total bisphenol A in human urine using solid-phase extraction and UPLC–MS/MS: method implementation, method qualification and troubleshooting

Journal of Chromatography B, 2015

The aim of the presented investigation was to document challenges encountered during implementati... more The aim of the presented investigation was to document challenges encountered during implementation and qualification of a method for bisphenol A (BPA) analysis and to develop and discuss precautions taken to avoid and to monitor contamination with BPA during sample handling and analysis. Previously developed and published HPLC-MS/MS methods for the determination of unconjugated BPA (Markham et al. Journal of Analytical Toxicology, 34 (2010) 293-303) [17] and total BPA (Markham et al. Journal of Analytical Toxicology, 38 (2014) 194-203) [20] in human urine were combined and transferred into another laboratory. The initial method for unconjugated BPA was developed and evaluated in two independent laboratories simultaneously. The second method for total BPA was developed and evaluated in one of these laboratories to conserve resources. Accurate analysis of BPA at sub-ppb levels is a challenging task as BPA is a widely used material and is ubiquitous in the environment at trace concentrations. Propensity for contamination of biological samples with BPA is reported in the literature during sample collection, storage, and/or analysis. Contamination by trace levels of BPA is so pervasive that even with extraordinary care, it is difficult to completely exclude the introduction of BPA into biological samples and, consequently, contamination might have an impact on BPA biomonitoring data. The applied UPLC-MS/MS method was calibrated from 0.05 to 25ng/ml. The limit of quantification was 0.1ng/ml for unconjugated BPA and 0.2ng/ml for total BPA, respectively, in human urine. Finally, the method was applied to urine samples derived from 20 volunteers. Overall, BPA can be analyzed in human urine with acceptable recovery and repeatability if sufficient measures are taken to avoid contamination throughout the procedure from sample collection until UPLC-MS/MS analysis.

Research paper thumbnail of Aryl hydrocarbon receptor knockout rats are insensitive to the pathological effects of repeated oral exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin

Journal of applied toxicology : JAT, Jan 17, 2015

Sustained activation of the aryl hydrocarbon receptor (AHR) is believed to be the initial key eve... more Sustained activation of the aryl hydrocarbon receptor (AHR) is believed to be the initial key event in AHR receptor-mediated tumorigenesis in the rat liver. The role of AHR in mediating pathological changes in the liver prior to tumor formation was investigated in a 4-week, repeated-dose study using adult female wild-type (WT) and AHR knockout (AHR-KO) rats treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Beginning at 8 weeks of age, AHR-KO and WT rats were dosed by oral gavage with varying concentrations of TCDD (0, 3, 22, 100, 300 and 1000 ng kg(-1) day(-1) ). Lung, liver and thymus histopathology, hematology, serum chemistry and the distribution of TCDD in liver and adipose tissue were examined. Treatment-related increases in the severity of liver and thymus pathology were observed in WT, but not AHR-KO rats. In the liver, these included hepatocellular hypertrophy, bile duct hyperplasia, multinucleated hepatocytes and inflammatory cell foci. A loss of cellularity in the ...

Research paper thumbnail of The adverse outcome pathway for rodent liver tumor promotion by sustained activation of the aryl hydrocarbon receptor

Regulatory Toxicology and Pharmacology, 2015

Research paper thumbnail of A Model for Aryl Hydrocarbon Receptor-Activated Gene Expression Shows Potency and Efficacy Changes and Predicts Squelching Due to Competition for Transcription Co-Activators

Research paper thumbnail of Mode of mutagenic action for the biocide Bioban CS-1246 in mouse lymphoma cells and implications for its in vivo mutagenic potential

Toxicological sciences : an official journal of the Society of Toxicology, 2005

The biocidal agent, BIOBAN CS-1246 (7-ethyl bicyclooxazolidine, CAS# 7747-35-5, CS-1246) induced ... more The biocidal agent, BIOBAN CS-1246 (7-ethyl bicyclooxazolidine, CAS# 7747-35-5, CS-1246) induced a concentration-dependent mutagenic response in mouse lymphoma (L5178Y TK+/-) cells both with and without the addition of S9 metabolic activation. Previous data indicating the ability of CS-1246 to hydrolyze in aqueous media to generate formaldehyde (FA), led us to investigate the potential role of FA in the CS-1246-induced mutagenic response in the mouse lymphoma assay (MLA). To accomplish this, the MLA on CS-1246 was repeated in the presence of a metabolizing system (formaldehyde dehydrogenase/NAD+), which was shown to successfully inhibit the mutagenic response of formaldehyde in this assay system. Significantly, the observed mutagenicity of CS-1246 was completely abrogated when the cultures were supplemented with formaldehyde dehydrogenase/NAD+, suggesting that the positive MLA response was attributable to the generation of FA in situ. These results demonstrate that in vitro mutageni...

Research paper thumbnail of Proposing a scientific confidence framework to help support the application of adverse outcome pathways for regulatory purposes

Regulatory Toxicology and Pharmacology, 2015

Research paper thumbnail of Background concentrations of dioxins, furans, and PCBs in Sprague-Dawley rats and juvenile swine

Journal of toxicology and environmental health. Part A, Jan 11, 2004

In preparation for a study of the relative oral bioavailability of polychlorinated dibenzo-p-diox... more In preparation for a study of the relative oral bioavailability of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) in soils (typically containing less than 1 ppb 2,3,7,8-tetrachlo-rodibenzo-p-dioxin [TCDD] toxic equivalents [TEQ]), the background concentrations of PCDD/Fs and selected polychlorinated biphenyls (PCBs) were measured in liver and adipose tissue from female Sprague-Dawley rats and juvenile swine after 30 d of ingesting laboratory chow. The measured concentrations of TCDD and other PCDD/Fs in rat livers were severalfold less than previously reported in the literature for control (unexposed) laboratory rodents. The concentrations of PCDD/Fs and selected PCBs in livers of swine were three- to fourfold lower than those reported for rats. The lower concentrations found in this study compared to previous findings may be due to inadvertent laboratory contamination in previous studies or to declining levels of PCDD/Fs in laboratory feed, which parallel the decline...

Research paper thumbnail of WE RECOGNIZE AND THANK THE FOLLOWING AS SUSTAINING MEMBERS FOR THE YEAR 2009

Research paper thumbnail of Sex ratio of the offspring of Sprague–Dawley rats exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in utero and lactationally in a three-generation study

Toxicology and Applied Pharmacology, 2006

Reports of a decreased male/female sex ratio in children born to males exposed to TCDD in Seveso,... more Reports of a decreased male/female sex ratio in children born to males exposed to TCDD in Seveso, Italy, at a young age have sparked examinations of this endpoint in other populations exposed to TCDD or related compounds. Overall, the male/female sex ratio results reported in these studies, with slightly different age-exposed male populations, have shown mixed results. Experimental studies of the effects of in utero exposure to TCDD in laboratory animals have reported no effect on the f(1) sex ratio and mixed results for the sex ratio of the f(2) generation. In order to better understand the potential effects of TCDD on second generation sex ratio, we retrieved archived data from a comprehensive three-generation feeding study of TCDD in rats that was conducted and published in the 1970s, but which did not publish data on sex ratio of the offspring [Murray, F.J., Smith, F.A., Nitschke, K.D., Humiston, C.G., Kociba, R.J., Schwetz, B.A., 1979. Three-generation reproduction study of rats given 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in the diet. Toxicol. Appl. Pharmacol. 50, 241-252]. A re-examination of the original Murray et al. data found no statistically significant treatment-related changes in postnatal day 1 sex ratio in any generation of treated animals, consistent with one other relatively large study reporting on this endpoint. We discuss mechanistic data underlying a potential effect of TCDD on this endpoint. We conclude that the inconsistency in findings on sex ratio of the offspring of male rats exposed to TCDD in utero is likely due to random variation associated with a relatively small sample size, although differences between studies in strain of rat, dose regimen, and day of ascertainment of sex ratio cannot be ruled out.

Research paper thumbnail of Recommended Relative Potency Factors for 2,3,4,7,8-Pentachlorodibenzofuran: The Impact of Different Dose Metrics

Toxicological Sciences, 2006

The recent National Toxicology Program (NTP) cancer bioassays for 2,3,7,8-tetrachlorodibenzo-p-di... more The recent National Toxicology Program (NTP) cancer bioassays for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and 2,3,4,7,8-pentachlorodibenzofuran (4-PeCDF) permit a reevaluation of the current TEF value of 4-PeCDF. The data also allow for the derivation of relative potency factors (RPFs) for cancer, which are based not only on administered dose but also on potentially more informative dose metrics, such as liver concentration, area under the liver concentration curve, and lifetime average body burden. Our analyses of these data indicate that chi-squared tests of observed versus predicted liver tumor incidence for 4-PeCDF reject the current TEF value of 0.5 value as too high. 4-PeCDF RPFs were derived using estimation methods that either did or did not assume parallelism of the 4-PeCDF and TCDD dose-response curves. The resulting parallelism-based RPFs for administered dose, liver concentration at terminal sacrifice, liver concentration AUC, and lifetime average body burden are 0.26, 0.014, 0.021, and 0.036, respectively. The administered dose RPF estimate is approximately one-half the current TEF value of 0.5. However, the use of administered dose fails to take into account pharmacokinetic differences between congeners and the generally acknowledged belief that body burden or some other measure of cumulative dose is more appropriate for estimating the health risk posed by persistent chemicals. The other three dose metrics do account for these important factors, and the corresponding RPFs are at least 10-fold lower than the current TEF for 4-PeCDF. In summary, our analyses support an administered dose TEF no greater than 0.25 and one in the 0.05-0.1 range for internal dose metrics such as lifetime average liver concentration or body burden.

Research paper thumbnail of Toxicokinetics Of 2,3,7,8-TCDF and 2,3,4,7,8-PeCDF in Mink (Mustela vison) at Ecologically Relevant Exposures

Toxicological Sciences, 2008

Research paper thumbnail of Comparative Temporal Toxicogenomic Analysis of TCDD- and TCDF-Mediated Hepatic Effects in Immature Female C57BL/6 Mice

Toxicological Sciences, 2008

Research paper thumbnail of Estimates of Cancer Potency of 2,3,7,8-Tetrachlorodibenzo(p)dioxin Using Linear and Nonlinear Dose-Response Modeling and Toxicokinetics

Toxicological Sciences, 2009

Research paper thumbnail of Estimates of Cancer Potency of 2,3,4,7,8-Pentachlorodibenzofuran Using Both Nonlinear and Linear Approaches

Toxicological Sciences, 2008

Cancer potency estimates were derived for 2,3,4,7,8-pentachlorodibenzofuran (4-PeCDF) using data ... more Cancer potency estimates were derived for 2,3,4,7,8-pentachlorodibenzofuran (4-PeCDF) using data collected from the recently published National Toxicology Program bioassay in female Sprague-Dawley rats. By using a toxicokinetic model for 4-PeCDF, the dose-response relationship for combined liver tumors (hepatocellular adenomas and cholangiocarcinomas) in rats was assessed in terms of lifetime average liver concentration and lifetime average adipose concentration with data from both the lifetime and the stop-exposure components of the bioassay. Benchmark dose modeling was performed to estimate tissue concentrations at two points of departure (EC(10) and EC(01) and their 95% upper and lower confidence limits). The same toxicokinetic model with human input values was then used to back-extrapolate human equivalent doses that corresponded to the internal tissue concentration measures at the points of departure. Information regarding the cancer mode of action was used to support the development of several toxicity criterion values based on a nonlinear method, e.g., reference dose or tolerable daily intake. Nonlinear estimates of toxicity criteria based on observed noncancer toxic events as possible precursors to tumor formation were also derived and were similar in value to those based on combined liver tumors. For comparison purposes, linear estimates of cancer potency were also derived.

Research paper thumbnail of Automated Dose-Response Analysis of the Relative Hepatic Gene Expression Potency of TCDF in C57BL/6 Mice

Toxicological Sciences, 2009

Toxic equivalency factors (TEFs) are assigned to dioxin-like chemicals based on relative potency ... more Toxic equivalency factors (TEFs) are assigned to dioxin-like chemicals based on relative potency (REP) values of individual adaptive and toxic responses compared to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Agilent 4x44K oligonucleotide microarrays were used to examine the hepatic gene expression potency of 2,3,7,8-tetrachlorodibenzofuran (TCDF), relative to TCDD with complementary histopathology, TCDD and TCDF tissue level analysis, and ethoxyresorufin-O-deethylase (EROD) assay data. Immature ovariectomized C57BL/6 mice were gavaged with 0.03, 0.1, 0.3, 1, 3, 10, 30, or 100 microg/kg TCDD, the World Health Organization TEF-adjusted doses (10 x TCDD dose) of TCDF (0.3, 1, 3, 10, 30, 100, or 300 microg/kg), or sesame oil vehicle and killed at 72 h. Two thousand two hundred eighty-eight and 1347 genes were differentially expressed (P1(t) > 0.90) at one or more doses by TCDD and TCDF, respectively. Automated dose-response modeling (ToxResponse Modeler) identified a total of 1027 and 837 genes with either a sigmoidal, exponential, linear, Gaussian, or quadratic dose-response relationship 72 h after treatment in TCDD and TCDF, respectively. Two hundred seventy genes exhibited a sigmoidal TCDD-induced dose-response (ED(50s) from 0.08 to 42.2 microg/kg) compared to only 179 sigmoidal responsive genes (ED(50s) from 0.74 to 299.9 microg/kg) elicited by TCDF. Of the 1027 TCDD dose-responsive genes, 654 were not examined further due to the lack of a dose response elicited by TCDF. Of the 373 genes that exhibited a TCDD and TCDF dose response, REPs were calculated for the 83 genes that exhibited comparable sigmoidal curve shapes and slopes. The median REP for these 83 genes was 0.10, with a maximum REP of 0.56 and a minimum of 0.01. REPs of 0.04 were also calculated for EROD and increase in relative liver weight (RLW) at 72 h. Collectively, the lower number of TCDF-induced genes compared to TCDD and the 0.04 REPs for EROD activity and increased RLW are not consistent with the TEF of 0.10 for the hepatotoxicity of TCDF in C57BL/6 mice at 72 h.

Research paper thumbnail of Automated Dose-Response Analysis and Comparative Toxicogenomic Evaluation of the Hepatic Effects Elicited by TCDD, TCDF, and PCB126 in C57BL/6 Mice

Toxicological Sciences, 2010

Research paper thumbnail of An Evaluation of Modeled Benzene Exposure and Dose Estimates Published in the Chinese–National Cancer Institute Collaborative Epidemiology Studies

Regulatory Toxicology and Pharmacology, 1999

Risk estimates and cause and effect determinations are directly dependent on exposure and dose-re... more Risk estimates and cause and effect determinations are directly dependent on exposure and dose-response relationships. Recently, relative risks and excess cancer mortality attributed to occupational benzene exposure have been published in collaborative studies conducted by Chinese investigators and scientists from the National Cancer Institute. The results of these studies suggest increased risk of acute nonlymphocytic leukemia at relatively low benzene concentrations and associations with cancers not previously associated with benzene exposure. These studies are potentially important due to their size and potential to more thoroughly investigate the link between benzene exposure and cancer. However, there are questions concerning the validity of exposure and dose estimates supporting relative risk characterizations in these studies. Apparent discrepancies between modeled exposure and dose estimates and sources of actual measured exposure information and clinical markers of benzene toxicity raise serious concerns questioning the reliability of relative risk and cancer associations stated in these studies.

Research paper thumbnail of The human AHR: identification of single nucleotide polymorphisms from six ethnic populations

Pharmacogenetics and Genomics, 2010

The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related dioxin-like chemicals are mediated... more The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related dioxin-like chemicals are mediated through binding-dependent activation of the cytosolic aryl hydrocarbon receptor (AHR). The human AHR is a low-affinity receptor relative to most rodents, but some reports suggest that there may be individuals with polymorphic high-affinity receptors, thereby possibly increasing the sensitivity to dioxins in such people. Although no polymorphisms have been reported in the ligand binding region of the AHR in the over 100 reported sequences, we sequenced 108 additional human AHR genes in an effort to further identify single single nucleotide polymorphisms (SNPs) within the open reading frames of the AHR locus. The DNA was sequenced from six ethnic populations that included Japanese, Chinese, European/Caucasian, African-American, South East Asian, and Hispanic. Six exonic SNPs were identified; four had been described as previously reported and two seem to be novel. Four of the SNPs identified lead to amino acid changes in the AHR protein and two of the SNPs lead to synonymous substitutions. An additional four SNPs have been reported elsewhere that were not identified in the current analysis. With these new sequences, more than 200 human AHR gene sequences have been analyzed for SNPs. The results indicate a very limited presence of polymorphisms in the core ligand binding region of the human AHR. Other regions, such as the transactivation domain, seem to be slightly more polymorphic in the human population and the impact on functionality should be further examined.

Research paper thumbnail of RAPID COMMUNICATION: BACKGROUND CONCENTRATIONS OF DIOXINS, FURANS, AND PCBs IN SPRAGUE-DAWLEY RATS AND JUVENILE SWINE

Journal of Toxicology and Environmental Health, Part A, 2004

... Michael V. Ruby,1 Stan W. Casteel,2 Timothy J. Evans,2 Kurt A. Fehling,3 Dennis J. Paustenbac... more ... Michael V. Ruby,1 Stan W. Casteel,2 Timothy J. Evans,2 Kurt A. Fehling,3 Dennis J. Paustenbach,3 Robert A. Budinsky,4 John P. Giesy,5 Lesa L. Aylward,6 Bryce D. Landenberger4 1Exponent, Inc ... We appreciate the assistance of Dr. Hon-Wing Leung and Terri Horvath. ...

Research paper thumbnail of AH receptor agonist activity in human blood measured with a cell-based bioassay: Evidence for naturally occurring AH receptor ligands in vivo

Journal of Exposure Science and Environmental Epidemiology, 2008

Research paper thumbnail of Quantitative analysis of unconjugated and total bisphenol A in human urine using solid-phase extraction and UPLC–MS/MS: method implementation, method qualification and troubleshooting

Journal of Chromatography B, 2015

The aim of the presented investigation was to document challenges encountered during implementati... more The aim of the presented investigation was to document challenges encountered during implementation and qualification of a method for bisphenol A (BPA) analysis and to develop and discuss precautions taken to avoid and to monitor contamination with BPA during sample handling and analysis. Previously developed and published HPLC-MS/MS methods for the determination of unconjugated BPA (Markham et al. Journal of Analytical Toxicology, 34 (2010) 293-303) [17] and total BPA (Markham et al. Journal of Analytical Toxicology, 38 (2014) 194-203) [20] in human urine were combined and transferred into another laboratory. The initial method for unconjugated BPA was developed and evaluated in two independent laboratories simultaneously. The second method for total BPA was developed and evaluated in one of these laboratories to conserve resources. Accurate analysis of BPA at sub-ppb levels is a challenging task as BPA is a widely used material and is ubiquitous in the environment at trace concentrations. Propensity for contamination of biological samples with BPA is reported in the literature during sample collection, storage, and/or analysis. Contamination by trace levels of BPA is so pervasive that even with extraordinary care, it is difficult to completely exclude the introduction of BPA into biological samples and, consequently, contamination might have an impact on BPA biomonitoring data. The applied UPLC-MS/MS method was calibrated from 0.05 to 25ng/ml. The limit of quantification was 0.1ng/ml for unconjugated BPA and 0.2ng/ml for total BPA, respectively, in human urine. Finally, the method was applied to urine samples derived from 20 volunteers. Overall, BPA can be analyzed in human urine with acceptable recovery and repeatability if sufficient measures are taken to avoid contamination throughout the procedure from sample collection until UPLC-MS/MS analysis.

Research paper thumbnail of Aryl hydrocarbon receptor knockout rats are insensitive to the pathological effects of repeated oral exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin

Journal of applied toxicology : JAT, Jan 17, 2015

Sustained activation of the aryl hydrocarbon receptor (AHR) is believed to be the initial key eve... more Sustained activation of the aryl hydrocarbon receptor (AHR) is believed to be the initial key event in AHR receptor-mediated tumorigenesis in the rat liver. The role of AHR in mediating pathological changes in the liver prior to tumor formation was investigated in a 4-week, repeated-dose study using adult female wild-type (WT) and AHR knockout (AHR-KO) rats treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Beginning at 8 weeks of age, AHR-KO and WT rats were dosed by oral gavage with varying concentrations of TCDD (0, 3, 22, 100, 300 and 1000 ng kg(-1) day(-1) ). Lung, liver and thymus histopathology, hematology, serum chemistry and the distribution of TCDD in liver and adipose tissue were examined. Treatment-related increases in the severity of liver and thymus pathology were observed in WT, but not AHR-KO rats. In the liver, these included hepatocellular hypertrophy, bile duct hyperplasia, multinucleated hepatocytes and inflammatory cell foci. A loss of cellularity in the ...

Research paper thumbnail of The adverse outcome pathway for rodent liver tumor promotion by sustained activation of the aryl hydrocarbon receptor

Regulatory Toxicology and Pharmacology, 2015

Research paper thumbnail of A Model for Aryl Hydrocarbon Receptor-Activated Gene Expression Shows Potency and Efficacy Changes and Predicts Squelching Due to Competition for Transcription Co-Activators

Research paper thumbnail of Mode of mutagenic action for the biocide Bioban CS-1246 in mouse lymphoma cells and implications for its in vivo mutagenic potential

Toxicological sciences : an official journal of the Society of Toxicology, 2005

The biocidal agent, BIOBAN CS-1246 (7-ethyl bicyclooxazolidine, CAS# 7747-35-5, CS-1246) induced ... more The biocidal agent, BIOBAN CS-1246 (7-ethyl bicyclooxazolidine, CAS# 7747-35-5, CS-1246) induced a concentration-dependent mutagenic response in mouse lymphoma (L5178Y TK+/-) cells both with and without the addition of S9 metabolic activation. Previous data indicating the ability of CS-1246 to hydrolyze in aqueous media to generate formaldehyde (FA), led us to investigate the potential role of FA in the CS-1246-induced mutagenic response in the mouse lymphoma assay (MLA). To accomplish this, the MLA on CS-1246 was repeated in the presence of a metabolizing system (formaldehyde dehydrogenase/NAD+), which was shown to successfully inhibit the mutagenic response of formaldehyde in this assay system. Significantly, the observed mutagenicity of CS-1246 was completely abrogated when the cultures were supplemented with formaldehyde dehydrogenase/NAD+, suggesting that the positive MLA response was attributable to the generation of FA in situ. These results demonstrate that in vitro mutageni...

Research paper thumbnail of Proposing a scientific confidence framework to help support the application of adverse outcome pathways for regulatory purposes

Regulatory Toxicology and Pharmacology, 2015

Research paper thumbnail of Background concentrations of dioxins, furans, and PCBs in Sprague-Dawley rats and juvenile swine

Journal of toxicology and environmental health. Part A, Jan 11, 2004

In preparation for a study of the relative oral bioavailability of polychlorinated dibenzo-p-diox... more In preparation for a study of the relative oral bioavailability of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) in soils (typically containing less than 1 ppb 2,3,7,8-tetrachlo-rodibenzo-p-dioxin [TCDD] toxic equivalents [TEQ]), the background concentrations of PCDD/Fs and selected polychlorinated biphenyls (PCBs) were measured in liver and adipose tissue from female Sprague-Dawley rats and juvenile swine after 30 d of ingesting laboratory chow. The measured concentrations of TCDD and other PCDD/Fs in rat livers were severalfold less than previously reported in the literature for control (unexposed) laboratory rodents. The concentrations of PCDD/Fs and selected PCBs in livers of swine were three- to fourfold lower than those reported for rats. The lower concentrations found in this study compared to previous findings may be due to inadvertent laboratory contamination in previous studies or to declining levels of PCDD/Fs in laboratory feed, which parallel the decline...

Research paper thumbnail of WE RECOGNIZE AND THANK THE FOLLOWING AS SUSTAINING MEMBERS FOR THE YEAR 2009

Research paper thumbnail of Sex ratio of the offspring of Sprague–Dawley rats exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in utero and lactationally in a three-generation study

Toxicology and Applied Pharmacology, 2006

Reports of a decreased male/female sex ratio in children born to males exposed to TCDD in Seveso,... more Reports of a decreased male/female sex ratio in children born to males exposed to TCDD in Seveso, Italy, at a young age have sparked examinations of this endpoint in other populations exposed to TCDD or related compounds. Overall, the male/female sex ratio results reported in these studies, with slightly different age-exposed male populations, have shown mixed results. Experimental studies of the effects of in utero exposure to TCDD in laboratory animals have reported no effect on the f(1) sex ratio and mixed results for the sex ratio of the f(2) generation. In order to better understand the potential effects of TCDD on second generation sex ratio, we retrieved archived data from a comprehensive three-generation feeding study of TCDD in rats that was conducted and published in the 1970s, but which did not publish data on sex ratio of the offspring [Murray, F.J., Smith, F.A., Nitschke, K.D., Humiston, C.G., Kociba, R.J., Schwetz, B.A., 1979. Three-generation reproduction study of rats given 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in the diet. Toxicol. Appl. Pharmacol. 50, 241-252]. A re-examination of the original Murray et al. data found no statistically significant treatment-related changes in postnatal day 1 sex ratio in any generation of treated animals, consistent with one other relatively large study reporting on this endpoint. We discuss mechanistic data underlying a potential effect of TCDD on this endpoint. We conclude that the inconsistency in findings on sex ratio of the offspring of male rats exposed to TCDD in utero is likely due to random variation associated with a relatively small sample size, although differences between studies in strain of rat, dose regimen, and day of ascertainment of sex ratio cannot be ruled out.

Research paper thumbnail of Recommended Relative Potency Factors for 2,3,4,7,8-Pentachlorodibenzofuran: The Impact of Different Dose Metrics

Toxicological Sciences, 2006

The recent National Toxicology Program (NTP) cancer bioassays for 2,3,7,8-tetrachlorodibenzo-p-di... more The recent National Toxicology Program (NTP) cancer bioassays for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and 2,3,4,7,8-pentachlorodibenzofuran (4-PeCDF) permit a reevaluation of the current TEF value of 4-PeCDF. The data also allow for the derivation of relative potency factors (RPFs) for cancer, which are based not only on administered dose but also on potentially more informative dose metrics, such as liver concentration, area under the liver concentration curve, and lifetime average body burden. Our analyses of these data indicate that chi-squared tests of observed versus predicted liver tumor incidence for 4-PeCDF reject the current TEF value of 0.5 value as too high. 4-PeCDF RPFs were derived using estimation methods that either did or did not assume parallelism of the 4-PeCDF and TCDD dose-response curves. The resulting parallelism-based RPFs for administered dose, liver concentration at terminal sacrifice, liver concentration AUC, and lifetime average body burden are 0.26, 0.014, 0.021, and 0.036, respectively. The administered dose RPF estimate is approximately one-half the current TEF value of 0.5. However, the use of administered dose fails to take into account pharmacokinetic differences between congeners and the generally acknowledged belief that body burden or some other measure of cumulative dose is more appropriate for estimating the health risk posed by persistent chemicals. The other three dose metrics do account for these important factors, and the corresponding RPFs are at least 10-fold lower than the current TEF for 4-PeCDF. In summary, our analyses support an administered dose TEF no greater than 0.25 and one in the 0.05-0.1 range for internal dose metrics such as lifetime average liver concentration or body burden.

Research paper thumbnail of Toxicokinetics Of 2,3,7,8-TCDF and 2,3,4,7,8-PeCDF in Mink (Mustela vison) at Ecologically Relevant Exposures

Toxicological Sciences, 2008

Research paper thumbnail of Comparative Temporal Toxicogenomic Analysis of TCDD- and TCDF-Mediated Hepatic Effects in Immature Female C57BL/6 Mice

Toxicological Sciences, 2008

Research paper thumbnail of Estimates of Cancer Potency of 2,3,7,8-Tetrachlorodibenzo(p)dioxin Using Linear and Nonlinear Dose-Response Modeling and Toxicokinetics

Toxicological Sciences, 2009

Research paper thumbnail of Estimates of Cancer Potency of 2,3,4,7,8-Pentachlorodibenzofuran Using Both Nonlinear and Linear Approaches

Toxicological Sciences, 2008

Cancer potency estimates were derived for 2,3,4,7,8-pentachlorodibenzofuran (4-PeCDF) using data ... more Cancer potency estimates were derived for 2,3,4,7,8-pentachlorodibenzofuran (4-PeCDF) using data collected from the recently published National Toxicology Program bioassay in female Sprague-Dawley rats. By using a toxicokinetic model for 4-PeCDF, the dose-response relationship for combined liver tumors (hepatocellular adenomas and cholangiocarcinomas) in rats was assessed in terms of lifetime average liver concentration and lifetime average adipose concentration with data from both the lifetime and the stop-exposure components of the bioassay. Benchmark dose modeling was performed to estimate tissue concentrations at two points of departure (EC(10) and EC(01) and their 95% upper and lower confidence limits). The same toxicokinetic model with human input values was then used to back-extrapolate human equivalent doses that corresponded to the internal tissue concentration measures at the points of departure. Information regarding the cancer mode of action was used to support the development of several toxicity criterion values based on a nonlinear method, e.g., reference dose or tolerable daily intake. Nonlinear estimates of toxicity criteria based on observed noncancer toxic events as possible precursors to tumor formation were also derived and were similar in value to those based on combined liver tumors. For comparison purposes, linear estimates of cancer potency were also derived.

Research paper thumbnail of Automated Dose-Response Analysis of the Relative Hepatic Gene Expression Potency of TCDF in C57BL/6 Mice

Toxicological Sciences, 2009

Toxic equivalency factors (TEFs) are assigned to dioxin-like chemicals based on relative potency ... more Toxic equivalency factors (TEFs) are assigned to dioxin-like chemicals based on relative potency (REP) values of individual adaptive and toxic responses compared to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Agilent 4x44K oligonucleotide microarrays were used to examine the hepatic gene expression potency of 2,3,7,8-tetrachlorodibenzofuran (TCDF), relative to TCDD with complementary histopathology, TCDD and TCDF tissue level analysis, and ethoxyresorufin-O-deethylase (EROD) assay data. Immature ovariectomized C57BL/6 mice were gavaged with 0.03, 0.1, 0.3, 1, 3, 10, 30, or 100 microg/kg TCDD, the World Health Organization TEF-adjusted doses (10 x TCDD dose) of TCDF (0.3, 1, 3, 10, 30, 100, or 300 microg/kg), or sesame oil vehicle and killed at 72 h. Two thousand two hundred eighty-eight and 1347 genes were differentially expressed (P1(t) > 0.90) at one or more doses by TCDD and TCDF, respectively. Automated dose-response modeling (ToxResponse Modeler) identified a total of 1027 and 837 genes with either a sigmoidal, exponential, linear, Gaussian, or quadratic dose-response relationship 72 h after treatment in TCDD and TCDF, respectively. Two hundred seventy genes exhibited a sigmoidal TCDD-induced dose-response (ED(50s) from 0.08 to 42.2 microg/kg) compared to only 179 sigmoidal responsive genes (ED(50s) from 0.74 to 299.9 microg/kg) elicited by TCDF. Of the 1027 TCDD dose-responsive genes, 654 were not examined further due to the lack of a dose response elicited by TCDF. Of the 373 genes that exhibited a TCDD and TCDF dose response, REPs were calculated for the 83 genes that exhibited comparable sigmoidal curve shapes and slopes. The median REP for these 83 genes was 0.10, with a maximum REP of 0.56 and a minimum of 0.01. REPs of 0.04 were also calculated for EROD and increase in relative liver weight (RLW) at 72 h. Collectively, the lower number of TCDF-induced genes compared to TCDD and the 0.04 REPs for EROD activity and increased RLW are not consistent with the TEF of 0.10 for the hepatotoxicity of TCDF in C57BL/6 mice at 72 h.

Research paper thumbnail of Automated Dose-Response Analysis and Comparative Toxicogenomic Evaluation of the Hepatic Effects Elicited by TCDD, TCDF, and PCB126 in C57BL/6 Mice

Toxicological Sciences, 2010

Research paper thumbnail of An Evaluation of Modeled Benzene Exposure and Dose Estimates Published in the Chinese–National Cancer Institute Collaborative Epidemiology Studies

Regulatory Toxicology and Pharmacology, 1999

Risk estimates and cause and effect determinations are directly dependent on exposure and dose-re... more Risk estimates and cause and effect determinations are directly dependent on exposure and dose-response relationships. Recently, relative risks and excess cancer mortality attributed to occupational benzene exposure have been published in collaborative studies conducted by Chinese investigators and scientists from the National Cancer Institute. The results of these studies suggest increased risk of acute nonlymphocytic leukemia at relatively low benzene concentrations and associations with cancers not previously associated with benzene exposure. These studies are potentially important due to their size and potential to more thoroughly investigate the link between benzene exposure and cancer. However, there are questions concerning the validity of exposure and dose estimates supporting relative risk characterizations in these studies. Apparent discrepancies between modeled exposure and dose estimates and sources of actual measured exposure information and clinical markers of benzene toxicity raise serious concerns questioning the reliability of relative risk and cancer associations stated in these studies.

Research paper thumbnail of The human AHR: identification of single nucleotide polymorphisms from six ethnic populations

Pharmacogenetics and Genomics, 2010

The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related dioxin-like chemicals are mediated... more The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related dioxin-like chemicals are mediated through binding-dependent activation of the cytosolic aryl hydrocarbon receptor (AHR). The human AHR is a low-affinity receptor relative to most rodents, but some reports suggest that there may be individuals with polymorphic high-affinity receptors, thereby possibly increasing the sensitivity to dioxins in such people. Although no polymorphisms have been reported in the ligand binding region of the AHR in the over 100 reported sequences, we sequenced 108 additional human AHR genes in an effort to further identify single single nucleotide polymorphisms (SNPs) within the open reading frames of the AHR locus. The DNA was sequenced from six ethnic populations that included Japanese, Chinese, European/Caucasian, African-American, South East Asian, and Hispanic. Six exonic SNPs were identified; four had been described as previously reported and two seem to be novel. Four of the SNPs identified lead to amino acid changes in the AHR protein and two of the SNPs lead to synonymous substitutions. An additional four SNPs have been reported elsewhere that were not identified in the current analysis. With these new sequences, more than 200 human AHR gene sequences have been analyzed for SNPs. The results indicate a very limited presence of polymorphisms in the core ligand binding region of the human AHR. Other regions, such as the transactivation domain, seem to be slightly more polymorphic in the human population and the impact on functionality should be further examined.

Research paper thumbnail of RAPID COMMUNICATION: BACKGROUND CONCENTRATIONS OF DIOXINS, FURANS, AND PCBs IN SPRAGUE-DAWLEY RATS AND JUVENILE SWINE

Journal of Toxicology and Environmental Health, Part A, 2004

... Michael V. Ruby,1 Stan W. Casteel,2 Timothy J. Evans,2 Kurt A. Fehling,3 Dennis J. Paustenbac... more ... Michael V. Ruby,1 Stan W. Casteel,2 Timothy J. Evans,2 Kurt A. Fehling,3 Dennis J. Paustenbach,3 Robert A. Budinsky,4 John P. Giesy,5 Lesa L. Aylward,6 Bryce D. Landenberger4 1Exponent, Inc ... We appreciate the assistance of Dr. Hon-Wing Leung and Terri Horvath. ...

Research paper thumbnail of AH receptor agonist activity in human blood measured with a cell-based bioassay: Evidence for naturally occurring AH receptor ligands in vivo

Journal of Exposure Science and Environmental Epidemiology, 2008