@article{hawkey_shekey_dean_asrat_koburov_holloway_kullman_levin_2024, title={Developmental exposure to pesticides that disrupt retinoic acid signaling causes persistent retinoid and behavioral dysfunction in zebrafish}, volume={1}, ISSN={["1096-0929"]}, DOI={10.1093/toxsci/kfae001}, abstractNote={Abstract}, journal={TOXICOLOGICAL SCIENCES}, author={Hawkey, Andrew B. and Shekey, Nathan and Dean, Cassandra and Asrat, Helina and Koburov, Reese and Holloway, Zade R. and Kullman, Seth W. and Levin, Edward D.}, year={2024}, month={Jan} }
 @article{wang_ritter_kullman_muddiman_2023, title={Comparative analysis of sucrose-embedding for whole-body zebrafish MSI by IR-MALDESI}, volume={8}, ISSN={["1618-2650"]}, url={https://doi.org/10.1007/s00216-023-04914-1}, DOI={10.1007/s00216-023-04914-1}, journal={ANALYTICAL AND BIOANALYTICAL CHEMISTRY}, author={Wang, Mary F. and Ritter, Morgan M. and Kullman, Seth W. and Muddiman, David C.}, year={2023}, month={Aug} }
 @article{phelps_palekar_conley_ferrero_driggers_linder_kullman_reif_sheats_dewitt_et al._2023, title={Legacy and emerging per- and polyfluoroalkyl substances suppress the neutrophil respiratory burst}, volume={20}, ISSN={["1547-6901"]}, url={https://doi.org/10.1080/1547691X.2023.2176953}, DOI={10.1080/1547691X.2023.2176953}, abstractNote={Abstract Per- and polyfluoroalkyl substances (PFASs) are used in a multitude of processes and products, including nonstick coatings, food wrappers, and fire-fighting foams. These chemicals are environmentally-persistent, ubiquitous, and can be detected in the serum of 98% of Americans. Despite evidence that PFASs alter adaptive immunity, few studies have investigated their effects on innate immunity. The report here presents results of studies that investigated the impact of nine environmentally-relevant PFASs [e.g. perfluorooctanoic acid (PFOA), perfluorooctane sulfonic acid potassium salt (PFOS-K), perfluorononanoic acid (PFNA), perfluorohexanoic acid (PFHxA), perfluorohexane sulfonic acid (PFHxS), perfluorobutane sulfonic acid (PFBS), ammonium perfluoro(2-methyl-3-oxahexanoate) (GenX), 7H-perfluoro-4-methyl-3,6-dioxa-octane sulfonic acid (Nafion byproduct 2), and perfluoromethoxyacetic acid sodium salt (PFMOAA-Na)] on one component of the innate immune response, the neutrophil respiratory burst. The respiratory burst is a key innate immune process by which microbicidal reactive oxygen species (ROS) are rapidly induced by neutrophils in response to pathogens; defects in the respiratory burst can increase susceptibility to infection. The study here utilized larval zebrafish, a human neutrophil-like cell line, and primary human neutrophils to ascertain whether PFAS exposure inhibits ROS production in the respiratory burst. It was observed that exposure to PFHxA and GenX suppresses the respiratory burst in zebrafish larvae and a human neutrophil-like cell line. GenX also suppressed the respiratory burst in primary human neutrophils. This report is the first to demonstrate that these PFASs suppress neutrophil function and support the utility of employing zebrafish larvae and a human cell line as screening tools to identify chemicals that may suppress human immune function.}, number={1}, journal={JOURNAL OF IMMUNOTOXICOLOGY}, author={Phelps, Drake W. and Palekar, Anika I. and Conley, Haleigh E. and Ferrero, Giuliano and Driggers, Jacob H. and Linder, Keith E. and Kullman, Seth W. and Reif, David M. and Sheats, M. Katie and DeWitt, Jamie C. and et al.}, year={2023}, month={Dec} }
 @article{joignant_ritter_knizner_garrard_kullman_muddiman_2023, title={Maximized Spatial Information and Minimized Acquisition Time of Top-Hat IR-MALDESI-MSI of Zebrafish Using Nested Regions of Interest (nROIs)}, volume={8}, ISSN={["1879-1123"]}, DOI={10.1021/jasms.3c00210}, abstractNote={Increasing the spatial resolution of a mass spectrometry imaging (MSI) method results in a more defined heatmap of the spatial distribution of molecules across a sample, but it is also associated with the disadvantage of increased acquisition time. Decreasing the area of the region of interest to achieve shorter durations results in the loss of potentially valuable information in larger specimens. This work presents a novel MSI method to reduce the time of MSI data acquisition with variable step size imaging: nested regions of interest (nROIs). Using nROIs, a small ROI may be imaged at a higher spatial resolution while nested inside a lower-spatial-resolution peripheral ROI. This conserves the maximal spatial and chemical information generated from target regions while also decreasing the necessary acquisition time. In this work, the nROI method was characterized on mouse liver and applied to top-hat MSI of zebrafish using a novel optical train, which resulted in a significant improvement in both acquisition time and spatial detail of the zebrafish. The nROI method can be employed with any step size pairing and adapted to any method in which the acquisition time of larger high-resolution ROIs poses a practical challenge.}, journal={JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY}, author={Joignant, Alena N. and Ritter, Morgan M. and Knizner, Kevan T. and Garrard, Kenneth P. and Kullman, Seth W. and Muddiman, David C.}, year={2023}, month={Aug} }
 @article{hamm_mahapatra_knuth_abedini_lingerfelt_ekins_kullman_2022, title={Confirmation of high-throughput screening data and novel mechanistic insights into FXR-xenobiotic interactions by orthogonal assays}, volume={3}, ISSN={["2666-027X"]}, DOI={10.1016/j.crtox.2022.100092}, abstractNote={Toxicology in the 21st Century (Tox21) is a federal collaboration employing a high-throughput robotic screening system to test 10,000 environmental chemicals. One of the primary goals of the program is prioritizing toxicity evaluations through in vitro high-throughput screening (HTS) assays for large numbers of chemicals already in commercial use for which little or no toxicity data is available. Within the Tox21 screening program, disruption in nuclear receptor (NR) signaling represents a particular area of interest. Given the role of NR's in modulating a wide range of biological processes, alterations of their activity can have profound biological impacts. Farnesoid X receptor (FXR) is a member of the nuclear receptor superfamily that has demonstrated importance in bile acid homeostasis, glucose metabolism, lipid homeostasis and hepatic regeneration. In this study, we re-evaluated 24 FXR agonists and antagonists identified through Tox21 using select orthogonal assays. In transient transactivation assays, 7/8 putative agonists and 4/4 putative inactive compounds were confirmed. Likewise, we confirmed 9/12 antagonists tested. Using a mammalian two hybrid approach we demonstrate that both FXR agonists and antagonists facilitate FXRα-coregulator interactions suggesting that differential coregulator recruitment may mediate activation/repression of FXRα mediated transcription. Additionally, we tested the ability of select FXR agonists and antagonists to facilitate hepatic transcription of FXR gene targets Shp and Bsep in a teleost (Medaka) model. Through application of in vitro cell-based assays, in silico modeling and in vivo gene expressions, we demonstrated the molecular complexity of FXR:ligand interactions and confirmed the ability of diverse ligands to modulate FXRα, facilitate differential coregulator recruitment and activate/repress receptor-mediated transcription. Overall, we suggest a multiplicative approach to assessment of nuclear receptor function may facilitate a greater understanding of the biological and mechanistic complexities of nuclear receptor activities and further our ability to interpret broad HTS outcomes.}, journal={CURRENT RESEARCH IN TOXICOLOGY}, author={Hamm, Jon and Mahapatra, Debabrata and Knuth, Megan M. and Abedini, Jaleh and Lingerfelt, Mary and Ekins, Sean and Kullman, Seth W.}, year={2022} }
 @article{kassotis_lefauve_chiang_knuth_schkoda_kullman_2022, title={Nonylphenol Polyethoxylates Enhance Adipose Deposition in Developmentally Exposed Zebrafish}, volume={10}, DOI={10.3390/toxics10020099}, abstractNote={Alkylphenol polyethoxylates (APEOs), such as nonylphenol ethoxylates (NPEOs), are high-production-volume surfactants used in laundry detergents, hard-surface cleaners, pesticide formulations, textile production, oils, paints, and other products. NPEOs comprise −80% of the total production of APEOs and are widely reported across diverse environmental matrices. Despite a growing push for replacement products, APEOs continue to be released into the environment through wastewater at significant levels. Research into related nonionic surfactants from varying sources has reported metabolic health impacts, and we have previously demonstrated that diverse APEOs and alcohol polyethoxylates promote adipogenesis in the murine 3T3-L1 pre-adipocyte model. These effects appeared to be independent of the base alkylphenol and related to the ethoxylate chain length, though limited research has evaluated NPEO exposures in animal models. The goals of this study were to assess the potential of NPEOs to promote adiposity (Nile red fluorescence quantification) and alter growth and/or development (toxicity, length, weight, and energy expenditure) of developmentally exposed zebrafish (Danio rerio). We also sought to expand our understanding of the ability to promote adiposity through evaluation in human mesenchymal stem cells. Herein, we demonstrated consistent adipogenic effects in two separate human bone-marrow-derived mesenchymal stem cell models, and that nonylphenol and its ethoxylates promoted weight gain and increased adipose deposition in developmentally exposed zebrafish. Notably, across both cell and zebrafish models we report increasing adipogenic/obesogenic activity with increasing ethoxylate chain lengths up to maximums around NPEO-6 and then decreasing activity with the longest ethoxylate chain lengths. This research suggests metabolic health concerns for these common obesogens, suggesting further need to assess molecular mechanisms and better characterize environmental concentrations for human health risk assessments.}, number={2}, journal={Toxics}, author={Kassotis, Christopher D. and LeFauve, Matthew K. and Chiang, Yu-Ting Tiffany and Knuth, Megan M. and Schkoda, Stacy and Kullman, Seth W.}, year={2022}, month={Feb}, pages={99} }
 @article{kassotis_lefauve_chiang_knuth_schkoda_kullman_2022, title={Nonylphenol Polyethoxylates Enhance Adipose Deposition in Developmentally Exposed Zebrafish (vol 10, 99, 2022)}, volume={10}, ISSN={["2305-6304"]}, DOI={10.3390/toxics10070345}, abstractNote={In the original publication [...]}, number={7}, journal={TOXICS}, author={Kassotis, Christopher D. and LeFauve, Matthew K. and Chiang, Yu-Ting Tiffany and Knuth, Megan M. and Schkoda, Stacy and Kullman, Seth W.}, year={2022}, month={Jul} }
 @article{watson_carmona baez_jima_reif_ding_roberts_kullman_2022, title={TCDD alters essential transcriptional regulators of osteogenic differentiation in multipotent mesenchymal stem cells}, volume={11}, ISSN={["1096-0929"]}, url={https://doi.org/10.1093/toxsci/kfac120}, DOI={10.1093/toxsci/kfac120}, abstractNote={Abstract}, journal={TOXICOLOGICAL SCIENCES}, author={Watson, AtLee T. D. and Carmona Baez, Aldo and Jima, Dereje and Reif, David and Ding, Jun and Roberts, Reade and Kullman, Seth W.}, year={2022}, month={Nov} }
 @article{knuth_stutts_ritter_garrard_kullman_2021, title={Vitamin D deficiency promotes accumulation of bioactive lipids and increased endocannabinoid tone in zebrafish}, volume={62}, ISSN={["1539-7262"]}, url={https://doi.org/10.1016/j.jlr.2021.100142}, DOI={10.1016/j.jlr.2021.100142}, abstractNote={Vitamin D is well known for its traditional role in bone mineral homeostasis; however, recent evidence suggests that vitamin D also plays a significant role in metabolic control. This study served to investigate putative linkages between vitamin D deficiency (VDD) and metabolic disruption of bioactive lipids by MS imaging. Our approach employed infrared-matrix-assisted laser desorption electrospray ionization MS imaging for lipid metabolite profiling in 6-month-old zebrafish fed either a VDD or a vitamin D-sufficient (VDS) diet. Using a lipidomics pipeline, we found that VDD zebrafish had a greater abundance of bioactive lipids (N-acyls, endocannabinoids [ECs], diacylglycerols/triacylglycerols, bile acids/bile alcohols, and vitamin D derivatives) suggestive of increased EC tone compared with VDS zebrafish. Tandem MS was performed on several differentially expressed metabolites with sufficient ion abundances to aid in structural elucidation and provide additional support for MS annotations. To confirm activation of the EC pathways, we subsequently examined expression of genes involved in EC biosynthesis, metabolism, and receptor signaling in adipose tissue and liver from VDD and VDS zebrafish. Gene expression changes were congruent with increased EC tone, with VDD zebrafish demonstrating increased synthesis and metabolism of anandamide compared with VDS zebrafish. Taken together, our data suggest that VDD may promote accumulation of bioactive lipids and increased EC tone in zebrafish.}, journal={JOURNAL OF LIPID RESEARCH}, publisher={Elsevier BV}, author={Knuth, Megan M. and Stutts, Whitney L. and Ritter, Morgan M. and Garrard, Kenneth P. and Kullman, Seth W.}, year={2021} }
 @article{baldwin_bain_di giulio_kullman_rice_ringwood_hurk_2020, title={20th Pollutant Responses in Marine Organisms (PRIMO 20): Global issues and fundamental mechanisms caused by pollutant stress in marine and freshwater organisms}, volume={227}, ISSN={["1879-1514"]}, DOI={10.1016/j.aquatox.2020.105620}, abstractNote={The 20th Pollutant Responses in Marine Organisms (PRIMO 20) conference provided a forum for scientists from around the world to communicate novel toxicological research findings specifically focused on aquatic organisms, by combining applied and basic research at the intersection of environmental and mechanistic toxicology. The work highlighted in this special issue of Aquatic Toxicology, a special issue of Marine Environmental Research, and presented through posters and presentations, encompass important and emerging topics in freshwater and marine toxicology. This includes multiple types of emerging contaminants including microplastics and UV filtering chemicals. Other studies aimed to further our understanding of the effects of endocrine disrupting chemicals, pharmaceuticals, and personal care products. Further research presented in this virtual issue examined the interactive effects of chemicals and pathogens, while the final set of manuscripts demonstrates continuing efforts to combine traditional biomonitoring, data from -omic technologies, and modeling for use in risk assessment and management. An additional goal of PRIMO meetings is to address the link between environmental and human health. Several articles in this issue of Aquatic Toxicology describe the appropriateness of using aquatic organisms as models for human health, while the keynote speakers, as described in the editorial below, presented research that highlighted bioaccumulation of contaminants such as PFOS and mercury from fish to marine mammals and coastal human populations such as the Gullah/GeeChee near Charleston, South Carolina, USA.}, journal={AQUATIC TOXICOLOGY}, author={Baldwin, William S. and Bain, Lisa J. and Di Giulio, Richard and Kullman, Seth and Rice, Charles D. and Ringwood, Amy H. and Hurk, Peter}, year={2020}, month={Oct} }
 @article{oliveri_glazer_mahapatra_kullman_levin_2020, title={Developmental exposure of zebrafish to vitamin D receptor acting drugs and environmental toxicants disrupts behavioral function}, volume={81}, ISSN={["1872-9738"]}, DOI={10.1016/j.ntt.2020.106902}, abstractNote={Vitamin D receptor (VDR) signaling is important for optimal neurobehavioral development. Disruption of VDR signaling by environmental toxicants during early development might contribute to the etiology of behavioral dysfunction. In the current set of studies, we examined ten compounds known to affect VDR function in vitro for neurobehavioral effects in vivo in zebrafish. Zebrafish embryos were exposed to concentrations of the compounds in their water during the first 5 days post-fertilization. On day 5, the embryos were tested in an alternating light-dark locomotor assay using a computerized video tracking system. We found that most of the compounds produced significant changes in locomotor behavior in exposed zebrafish larvae, although the direction of the effect (i.e., hypo- or hyperactivity) and the sensitivity of the effect to changes in illumination condition varied across the compounds. The nature of the behavioral effects generally corresponded to the effects these compounds have been shown to exert on VDR. These studies lay a foundation for further investigation to determine whether behavioral dysfunction persists into adulthood and if so which behavioral functions are affected. Zebrafish can be useful for screening compounds identified in high throughput in vitro assays to provide an initial test for how those compounds would affect construction and behavioral function of a complex nervous system, helping to bridge the gap between in vitro neurotoxicity assays and mammalian models for risk assessment in humans.}, journal={NEUROTOXICOLOGY AND TERATOLOGY}, author={Oliveri, Anthony N. and Glazer, Lilah and Mahapatra, Debabrata and Kullman, Seth W. and Levin, Edward D.}, year={2020} }
 @article{stutts_knuth_ekelöf_mahapatra_kullman_muddiman_2020, title={Methods for Cryosectioning and Mass Spectrometry Imaging of Whole-Body Zebrafish}, volume={31}, ISSN={1044-0305 1879-1123}, url={http://dx.doi.org/10.1021/jasms.9b00097}, DOI={10.1021/jasms.9b00097}, abstractNote={The zebrafish (Danio rerio) is an ideal model for whole animal studies of lipid metabolism and lipid-related disease. In this work, infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) mass spectrometry imaging (MSI) was applied for direct visualization of lipid and metabolite distributions across various organs in whole-body zebrafish tissue sections. Detailed methods for overcoming the challenges of cryosectioning adult male zebrafish for MSI and complementary histological imaging are described. Representative two-dimensional ion maps demonstrated organ specific localization of lipid analytes allowing for visualization of areas of interest including the brain, liver, intestines, and skeletal muscle. A high resolving power mass spectrometer was utilized for accurate mass measurements, which permitted the use of open-source, web-based tools for MS1 annotations including METASPACE and METLIN. Whole-body MSI with IR-MALDESI allowed for broad lipid coverage with high spatial resolution, illustrating the potential of this technique for studying lipid-related diseases using zebrafish as a model organism.}, number={4}, journal={Journal of the American Society for Mass Spectrometry}, publisher={American Chemical Society (ACS)}, author={Stutts, Whitney L. and Knuth, Megan M. and Ekelöf, Måns and Mahapatra, Debabrata and Kullman, Seth W. and Muddiman, David C.}, year={2020}, month={Feb}, pages={768–772} }
 @article{kassotis_herkert_hammel_hoffman_xia_kullman_sosa_stapleton_2020, title={Thyroid Receptor Antagonism of Chemicals Extracted from Personal Silicone Wristbands within a Papillary Thyroid Cancer Pilot Study}, volume={54}, ISSN={["1520-5851"]}, DOI={10.1021/acs.est.0c05972}, abstractNote={Research suggests that thyroid cancer incidence rates are increasing, and environmental exposures have been postulated to be playing a role. To explore this possibility, we conducted a pilot study to investigate the thyroid disrupting bioactivity of chemical mixtures isolated from personal silicone wristband samplers within a thyroid cancer cohort. Specifically, we evaluated TRβ antagonism of chemical mixtures extracted from wristbands (n = 72) worn by adults in central North Carolina participating in a case-control study on papillary thyroid cancer. Sections of wristbands were solvent-extracted and analyzed via mass spectrometry to quantify a suite of semivolatile chemicals. A second extract from each wristband was used in a bioassay to quantify TRβ antagonism in human embryonic kidney cells (HEK293/17) at concentrations ranging from 0.1 to 10% of the original extract (by volume). Approximately 70% of the sample extracts tested at a 1% extract concentration exhibited significant TRβ antagonism, with a mean of 30% and a range of 0-100%. Inhibited cell viability was noted in >20% of samples that were tested at 5 and 10% concentrations. Antagonism was positively associated with wristband concentrations of several phthalates, organophosphate esters, and brominated flame retardants. These results suggest that personal passive samplers may be useful in evaluating the bioactivities of mixtures that people contact on a daily basis. We also report tentative associations between thyroid receptor antagonism, chemical concentrations, and papillary thyroid cancer case status. Future research utilizing larger sample sizes, prospective data collection, and measurement of serum thyroid hormone levels (which were not possible in this study) should be utilized to more comprehensively evaluate these associations.}, number={23}, journal={ENVIRONMENTAL SCIENCE & TECHNOLOGY}, author={Kassotis, Christopher D. and Herkert, Nicholas J. and Hammel, Stephanie C. and Hoffman, Kate and Xia, Qianyi and Kullman, Seth W. and Sosa, Julie Ann and Stapleton, Heather M.}, year={2020}, month={Dec}, pages={15296–15312} }
 @article{knuth_mahapatra_jima_wan_hammock_law_kullman_2020, title={Vitamin D deficiency serves as a precursor to stunted growth and central adiposity in zebrafish}, volume={10}, ISSN={["2045-2322"]}, DOI={10.1038/s41598-020-72622-2}, abstractNote={Abstract}, number={1}, journal={SCIENTIFIC REPORTS}, author={Knuth, Megan M. and Mahapatra, Debabrata and Jima, Dereje and Wan, Debin and Hammock, Bruce D. and Law, Mac and Kullman, Seth W.}, year={2020}, month={Sep} }
 @article{oliveri_knuth_glazer_bailey_kullman_levin_2020, title={Zebrafish show long-term behavioral impairments resulting from developmental vitamin D deficiency}, volume={224}, ISSN={["0031-9384"]}, DOI={10.1016/j.physbeh.2020.113016}, abstractNote={Vitamin D has been shown in a wide variety of species to play critical roles in neurodevelopment. Vitamin D deficiency disrupts development of the brain and can cause lasting behavioral dysfunction. Zebrafish have become an important model for the study of development in general and neurodevelopment in particular. Zebrafish were used in the current study to characterize the effects of developmental vitamin D deficiency on behavioral function. Adult zebrafish that had been chronically fed a vitamin D deficient or replete diets were bred and the offspring were continued on those diets. The offspring were behaviorally tested as adults. In the novel tank diving test the vitamin D deficient diet significantly lowered the vertical position of fish indicative of more anxiety-like behavior. In the novel tank diving test swimming activity was also significantly decreased by vitamin D deficiency. Startle response was increased by developmental vitamin D deficiency during the early part of the test. No significant effects of vitamin D deficiency were seen with social affiliation and predatory stimulus avoidance tests. These results indicate a phenotype of vitamin D deficiency characterized by more anxiety-like behavior. This result was relatively specific inasmuch as few or no behavioral effects were seen in other behavioral tests.}, journal={PHYSIOLOGY & BEHAVIOR}, author={Oliveri, Anthony N. and Knuth, Megan and Glazer, Lilah and Bailey, Jordan and Kullman, Seth W. and Levin, Edward D.}, year={2020}, month={Oct} }
 @inbook{kullman_watson_2019, place={Hoboken, NJ}, edition={2nd}, title={Alterations in skeletogenesis during medaka embryonic development}, ISBN={9781119575290 9781119575399}, booktitle={Medaka: Biology, Management, and Experimental Protocols}, publisher={Wiley}, author={Kullman, S.W. and Watson, A.D.}, editor={Murata, K. and Kinoshita, M. and Naruse, K. and Tanaka, M. and Kamei, Y.Editors}, year={2019} }
 @article{watson_nordberg_loboa_kullman_2019, title={Evidence for Aryl hydrocarbon Receptor-Mediated Inhibition of Osteoblast Differentiation in Human Mesenchymal Stem Cells}, volume={167}, ISSN={["1096-0929"]}, DOI={10.1093/toxsci/kfy225}, abstractNote={Multipotent mesenchymal stem cells (MSCs) maintain the ability to differentiate into adipogenic, chondrogenic, or osteogenic cell lineages. There is increasing concern that exposure to environmental agents such as aryl hydrocarbon receptor (AhR) ligands, may perturb the osteogenic pathways responsible for normal bone formation. The objective of the current study was to evaluate the potential of the prototypic AhR ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to disrupt osteogenic differentiation of human bone-derived MSCs (hBMSCs) in vitro. Primary hBMSCs from three donors were exposed to 10 nM TCDD and differentiation was interrogated using select histological, biochemical, and transcriptional markers of osteogenesis. Exposure to 10 nM TCDD resulted in an overall consistent attenuation of alkaline phosphatase (ALP) activity and matrix mineralization at terminal stages of differentiation in primary hBMSCs. At the transcriptional level, the transcriptional regulator DLX5 and additional osteogenic markers (ALP, OPN, and IBSP) displayed attenuated expression; conversely, FGF9 and FGF18 were consistently upregulated in each donor. Expression of stem cell potency markers SOX2, NANOG, and SALL4 decreased in the osteogenic controls, whereas expression in TCDD-treated cells resembled that of undifferentiated cells. Coexposure with the AhR antagonist GNF351 blocked TCDD-mediated attenuation of matrix mineralization, and either fully or partially rescued expression of genes associated with osteogenic regulation, extracellular matrix, and/or maintenance of multipotency. Thus, experimental evidence from this study suggests that AhR transactivation likely attenuates osteoblast differentiation in multipotent hBMSCs. This study also underscores the use of primary human MSCs to evaluate osteoinductive or osteotoxic potential of chemical and pharmacologic agents in vitro.}, number={1}, journal={TOXICOLOGICAL SCIENCES}, author={Watson, AtLee T. D. and Nordberg, Rachel C. and Loboa, Elizabeth G. and Kullman, Seth W.}, year={2019}, month={Jan}, pages={145–156} }
 @misc{ideraabdullah_belenchia_rosenfeld_kullman_knuth_mahapatra_bereman_levinlo_peterson_2019, title={Maternal vitamin D deficiency and developmental origins of health and disease (DOHaD)}, volume={241}, ISSN={["1479-6805"]}, DOI={10.1530/JOE-18-0541}, abstractNote={Vitamin D is an essential nutrient that is metabolized in the body to generate an active metabolite (1,25(OH)2D) with hormone-like activity and highly diverse roles in cellular function. Vitamin D deficiency (VDD) is a prevalent but easily preventable nutritional disturbance. Emerging evidence demonstrates the importance of sufficient vitamin D concentrations during fetal life with deficiencies leading to long-term effects into adulthood. Here, we provide a detailed review and perspective of evidence for the role of maternal VDD in offspring long-term health, particularly as it relates to developmental origins of health and disease (DOHaD). We focus on the roles in neurobehavioral and cardiometabolic disorders in humans and highlight recent findings from zebrafish and rodent models that probe potential mechanisms linking early life VDD to later life health outcomes. Moreover, we explore evidence implicating epigenetic mechanisms as a mediator of this link. Gaps in our current understanding of how maternal VDD might result in deleterious offspring outcomes later in life are also addressed.}, number={2}, journal={JOURNAL OF ENDOCRINOLOGY}, author={Ideraabdullah, Folami Y. and Belenchia, Anthony M. and Rosenfeld, Cheryl S. and Kullman, Seth W. and Knuth, Megan and Mahapatra, Debabrata and Bereman, Michael and Levinlo, Edward D. and Peterson, Catherine A.}, year={2019}, month={May}, pages={R65–R80} }
 @article{lee_kullman_yost_worley-davis_reckhow_2018, title={An object-oriented Bayesian network approach for establishing swine manure-borne natural estrogenic compounds budget}, volume={639}, ISSN={["1879-1026"]}, DOI={10.1016/j.scitotenv.2018.05.209}, abstractNote={A facility-wide estrogen budget model was developed to assess the excretion of natural estrogens by swine in a commercial swine farrowing concentrated animal feeding operations (CAFO) in North Carolina, using an object-oriented Bayesian network (OOBN) approach. The OOBN model is the combination of twelve objects of Bayesian network models, which characterize the estrogen budget flows based on the sow reproductive cycle (i.e., pre-estrus, estrus, and lactation) for the three natural estrogen types [estrone (E1), 17β-estradiol (E2), and estriol (E3)] within each barn. This OOBN model provides a mechanism to quantify the levels of the natural estrogens and their probabilistic distributions with regard to estrogen type, waste sources such as urine, feces, and recycling lagoon slurry, and animal reproductive status. Moreover, the OOBN model allows us to assess the overall contribution of natural estrogen compounds from each operational unit of the CAFO, while accounting for the uncertainties. Results from the OOBN model indicate a rank order of lactating sows > gestating sows > breeding sows in terms of contribution of estrogen loads to the total natural estrogen budget. As to estrogen type, E1 was found as the major estrogen metabolite with the summed concentrations of urine, feces, and flushing slurry wastes exceeding 3000 ng/l > 90% of the time. As to waste sources, the flushing slurry waste was found to be a major contributor of the estrogen budget compared with urine and feces wastes from barn animals.}, journal={SCIENCE OF THE TOTAL ENVIRONMENT}, author={Lee, Boknam and Kullman, Sethw. and Yost, Erin E. and Worley-Davis, Lynn and Reckhow, Kenneth H.}, year={2018}, month={Oct}, pages={815–825} }
 @article{mahapatra_franzosa_roell_kuenemann_houck_reif_fourches_kullman_2018, title={Confirmation of high-throughput screening data and novel mechanistic insights into VDR-xenobiotic interactions by orthogonal assays}, volume={8}, ISSN={2045-2322}, url={http://dx.doi.org/10.1038/S41598-018-27055-3}, DOI={10.1038/S41598-018-27055-3}, abstractNote={Abstract}, number={1}, journal={Scientific Reports}, publisher={Springer Science and Business Media LLC}, author={Mahapatra, Debabrata and Franzosa, Jill A. and Roell, Kyle and Kuenemann, Melaine Agnes and Houck, Keith A. and Reif, David M. and Fourches, Denis and Kullman, Seth W.}, year={2018}, month={Jun} }
 @article{neelon_white_vidal_schildkraut_murtha_murphy_kullman_hoyo_2018, title={Maternal vitamin D, DNA methylation at imprint regulatory regions and offspring weight at birth, 1 year and 3 years}, volume={42}, ISSN={["1476-5497"]}, DOI={10.1038/ijo.2017.160}, abstractNote={Vitamin D deficiency during pregnancy is associated with poor birth outcomes in some studies, but few have examined weight beyond birth. In addition, little is known about how vitamin D influences DNA methylation of regulatory regions known to be involved in growth, as possible mediators to weight status in offspring. We conducted linear regressions to assess maternal plasma 25-hydroxyvitamin D (25(OH)D) by quartile and birth weight for gestational age z-score, 1-year weight-for-length z-score and 3-year body mass index (BMI) z-score among 476 mother/infant dyads from a prospective cohort. We assessed maternal 25(OH)D and infant DNA methylation at nine differentially methylated regions (DMRs) of genomically imprinted genes with known functions in fetal growth, including H19, IGF2, MEG3, MEG3-IG, MEST, NNAT, PEG3, PLAGL1 and SGCE/PEG10. Mean (standard deviation, s.d.) maternal 25(OH)D was 41.1 (14.2) nmol l−m at a mean (s.d.) of 13.2 (5.5) weeks gestation. After adjustment for potential confounders, the first (Q1) and second (Q2) quartiles of 25(OH)D, compared to the fourth (Q4), were associated with lower birth weight for gestational age z-scores (−0.43 units; CI: −0.79, −0.07; P=0.02 for Q1 and −0.56 units; CI: −0.89, −0.23; P=0.001 for Q2). Q1 compared to Q4 was associated with higher 1-year weight-for-length z-scores (0.78 units; 0.08, 1.54; P=0.04) and higher 3-year BMI z-scores (0.83 units; 0.11, 0.93; P=0.02). We did not observe associations between maternal 25(OH)D and methylation for any of the nine DMRs after correcting for multiple testing. Reduced maternal 25(OH)D was associated with lower birth weight for gestational age z-scores but higher 1-year weight-for-length and 3-year BMI z-scores in offspring. However, 25(OH)D does not appear to be operating through the regulatory sequences of the genomically imprinted genes we examined.}, number={4}, journal={INTERNATIONAL JOURNAL OF OBESITY}, author={Neelon, S. E. Benjamin and White, A. J. and Vidal, A. C. and Schildkraut, J. M. and Murtha, A. P. and Murphy, S. K. and Kullman, S. W. and Hoyo, C.}, year={2018}, month={Apr}, pages={587–593} }
 @inbook{kullman_baldwin_leblanc_2018, place={Hoboken, NJ}, edition={5th}, title={Nuclear receptors}, booktitle={Molecular and Biochemical Toxicology}, publisher={John Wiley & Sons}, author={Kullman, S.W. and Baldwin, W.B. and LeBlanc, G.A.}, editor={Smart, Robert C. and Hodgson, ErnestEditors}, year={2018}, pages={293–326} }
 @article{lee_kullman_yost_meyer_worley-davis_williams_reckhow_2018, title={Predicting characteristics of rainfall driven estrogen runoff and transport from swine AFO spray fields (vol 532, pg 571, 2015)}, volume={628-629}, ISSN={["1879-1026"]}, DOI={10.1016/j.scitotenv.2018.02.141}, journal={SCIENCE OF THE TOTAL ENVIRONMENT}, author={Lee, Boknam and Kullman, Seth W. and Yost, Erin E. and Meyer, Michael T. and Worley-Davis, Lynn and Williams, C. Michael and Reckhow, Kenneth H.}, year={2018}, month={Jul}, pages={1460–1460} }
 @article{grieshaber_penland_kwak_cope_heise_law_shea_aday_rice_kullman_et al._2018, title={Relation of contaminants to fish intersex in riverine sport fishes}, volume={643}, ISSN={0048-9697}, url={http://dx.doi.org/10.1016/J.SCITOTENV.2018.06.071}, DOI={10.1016/j.scitotenv.2018.06.071}, abstractNote={Endocrine active compounds (EACs) are pollutants that have been recognized as an emerging and widespread threat to aquatic ecosystems globally. Intersex, the presence of female germ cells within a predominantly male gonad, is considered a biomarker of endocrine disruption caused by EACs. We measured a suite of EACs and assessed their associated impacts on fish intersex occurrence and severity in a large, regulated river system in North Carolina and South Carolina, USA. Our specific objective was to determine the relationship of contaminants in water, sediment, and fish tissue with the occurrence and severity of the intersex condition in wild, adult black bass (Micropterus), sunfish (Lepomis), and catfish (Ictaluridae) species at 11 sites located on the Yadkin-Pee Dee River. Polycyclic aromatic hydrocarbons (PAHs), ethinylestradiol (EE2), and heavy metals were the most prevalent contaminants that exceeded effect levels for the protection of aquatic organisms. Fish intersex condition was most frequently observed and most severe in black basses and was less frequently detected and less severe in sunfishes and catfishes. The occurrence of the intersex condition in fish showed site-related effects, rather than increasing longitudinal trends from upstream to downstream. Mean black bass and catfish tissue contaminant concentrations were higher than that of sunfish, likely because of the latter's lower trophic position in the food web. Principal component analysis identified waterborne PAHs as the most correlated environmental contaminant with intersex occurrence and severity in black bass and sunfish. As indicated by the intersex condition, EACs have adverse but often variable effects on the health of wild sport fishes in this river, likely due to fluctuations in EAC inputs and the dynamic nature of the riverine system. These findings enhance the understanding of the relationship between contaminants and fish health and provide information to guide ecologically comprehensive conservation and management decisions.}, journal={Science of The Total Environment}, publisher={Elsevier BV}, author={Grieshaber, C.A. and Penland, T.N. and Kwak, T.J. and Cope, W.G. and Heise, R.J. and Law, J.M. and Shea, Damian and Aday, D.D. and Rice, J.A. and Kullman, S.W. and et al.}, year={2018}, month={Dec}, pages={73–89} }
 @article{lee pow_law_kwak_cope_rice_kullman_aday_2017, title={Endocrine active contaminants in aquatic systems and intersex in common sport fishes}, volume={36}, ISSN={0730-7268}, url={http://dx.doi.org/10.1002/ETC.3607}, DOI={10.1002/etc.3607}, abstractNote={Abstract}, number={4}, journal={Environmental Toxicology and Chemistry}, publisher={Wiley}, author={Lee Pow, Crystal S.D. and Law, J. Mac and Kwak, Thomas J. and Cope, W. Gregory and Rice, James A. and Kullman, Seth W. and Aday, D. Derek}, year={2017}, month={Apr}, pages={959–968} }
 @article{macaulay_chernick_chen_hinton_bailey_kullman_levin_stapleton_2017, title={Exposure to a PBDE/OH-BDE mixture alters juvenile zebrafish (Danio rerio) development}, volume={36}, ISSN={["1552-8618"]}, DOI={10.1002/etc.3535}, abstractNote={Abstract}, number={1}, journal={ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY}, author={Macaulay, Laura J. and Chernick, Melissa and Chen, Albert and Hinton, David E. and Bailey, Jordan M. and Kullman, Seth W. and Levin, Edward D. and Stapleton, Heather M.}, year={2017}, month={Jan}, pages={36–48} }
 @article{lee pow_tilahun_creech_law_cope_kwak_rice_aday_kullman_2017, title={Windows of Susceptibility and Consequences of Early Life Exposures to 17β–estradiol on Medaka (<i>Oryzias latipes</i>) Reproductive Success}, volume={51}, ISSN={0013-936X 1520-5851}, url={http://dx.doi.org/10.1021/acs.est.7b01568}, DOI={10.1021/acs.est.7b01568}, abstractNote={Estrogens and estrogen mimics are commonly found in surface waters and are associated with deleterious effects in fish populations. Impaired fertility and fecundity in fish following chronic exposures to estrogens and estrogen mimics during critical windows in development are well documented. However, information regarding differential reproductive effects of exposure within defined developmental stages remains sparse. In this study, reproductive capacity was assessed in Japanese medaka (Oryzias latipes) after exposure to two concentrations of 17β-estradiol (E2β; 2 ng/L and 50 ng/L) during four distinct stages of development: gonad development, gonad differentiation, development of secondary sex characteristics (SSC) and gametogenesis. Exposure to E2β did not adversely impact survival, hatch success, growth, or genotypic ratios. In contrast, exposure to 50 ng/L E2β during SSC development altered phenotypic ratios and SSC. Exposure to both E2β treatments reduced reproductive capacity (fertility, fecundity) by 7.3-57.4% in adult medaka breeding pairs, with hindrance of SSC development resulting in the largest disruption in breeding capacity (51.6-57.4% decrease) in the high concentration. This study documents differential effects among four critical stages of development and provides insight into factors (window of exposure, exposure concentration and duration of exposure period) contributing to reproductive disruption in fish.}, number={9}, journal={Environmental Science & Technology}, publisher={American Chemical Society (ACS)}, author={Lee Pow, Crystal S.D. and Tilahun, Kedamawit and Creech, Kari and Law, J. Mac and Cope, W. Gregory and Kwak, Thomas J. and Rice, James A. and Aday, D. Derek and Kullman, Seth W.}, year={2017}, month={Apr}, pages={5296–5305} }
 @article{planchart_mattingly_allen_ceger_casey_hinton_kanungo_kullman_tal_bondesson_et al._2016, title={Advancing toxicology research using in vivo high throughput toxicology with small fish models}, volume={33}, ISSN={1868-596X}, url={http://dx.doi.org/10.14573/altex.1601281}, DOI={10.14573/altex.1601281}, abstractNote={Summary Small freshwater fish models, especially zebrafish, offer advantages over traditional rodent models, including low maintenance and husbandry costs, high fecundity, genetic diversity, physiology similar to that of traditional biomedical models, and reduced animal welfare concerns. The Collaborative Workshop on Aquatic Models and 21st Century Toxicology was held at North Carolina State University on May 5-6, 2014, in Raleigh, North Carolina, USA. Participants discussed the ways in which small fish are being used as models to screen toxicants and understand mechanisms of toxicity. Workshop participants agreed that the lack of standardized protocols is an impediment to broader acceptance of these models, whereas development of standardized protocols, validation, and subsequent regulatory acceptance would facilitate greater usage. Given the advantages and increasing application of small fish models, there was widespread interest in follow-up workshops to review and discuss developments in their use. In this article, we summarize the recommendations formulated by workshop participants to enhance the utility of small fish species in toxicology studies, as well as many of the advances in the field of toxicology that resulted from using small fish species, including advances in developmental toxicology, cardiovascular toxicology, neurotoxicology, and immunotoxicology. We also review many emerging issues that will benefit from using small fish species, especially zebrafish, and new technologies that will enable using these organisms to yield results unprecedented in their information content to better understand how toxicants affect development and health.}, number={4}, journal={ALTEX - Alternatives to animal experimentation,}, publisher={ALTEX Edition}, author={Planchart, Antonio and Mattingly, Carolyn J. and Allen, David and Ceger, Patricia and Casey, Warren and Hinton, David and Kanungo, Jyotshna and Kullman, Seth W. and Tal, Tamara and Bondesson, Maria and et al.}, year={2016}, pages={435–452} }
 @article{kollitz_zhang_hawkins_whitfield_reif_kullman_2016, title={Evolutionary and Functional Diversification of the Vitamin D Receptor-Lithocholic Acid Partnership}, volume={11}, ISSN={["1932-6203"]}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-85005987339&partnerID=MN8TOARS}, DOI={10.1371/journal.pone.0168278}, abstractNote={The evolution, molecular behavior, and physiological function of nuclear receptors are of particular interest given their diverse roles in regulating essential biological processes. The vitamin D receptor (VDR) is well known for its canonical roles in calcium homeostasis and skeletal maintenance. Additionally, VDR has received an increased amount of attention due to the discovery of numerous non-calcemic functions, including the detoxification of lithocholic acid. Lithocholic acid is a toxic metabolite of chenodeoxycholic acid, a primary bile acid. The partnership between the VDR and lithocholic acid has been hypothesized to be a recent adaptation that evolved to mediate the detoxification and elimination of lithocholic acid from the gut. This partnership is speculated to be limited to higher vertebrates (birds and mammals), as lower vertebrates do not synthesize the parent compound of lithocholic acid. However, the molecular functions associated with the observed insensitivity of basal VDRs to lithocholic acid have not been explored. Here we characterize canonical nuclear receptor functions of VDRs from select species representing key nodes in vertebrate evolution and span a range of bile salt phenotypes. Competitive ligand binding assays revealed that the receptor’s affinity for lithocholic acid is highly conserved across species, suggesting that lithocholic acid affinity is an ancient and non-adaptive trait. However, transient transactivation assays revealed that lithocholic acid-mediated VDR activation might have evolved more recently, as the non-mammalian receptors did not respond to lithocholic acid unless exogenous coactivator proteins were co-expressed. Subsequent functional assays indicated that differential lithocholic acid-mediated receptor activation is potentially driven by differential protein-protein interactions between VDR and nuclear receptor coregulator proteins. We hypothesize that the vitamin D receptor-lithocholic acid partnership evolved as a by-product of natural selection on the ligand-receptor partnership between the vitamin D receptor and the native VDR ligand: 1α,25-dihydroxyvitamin D3, the biologically active metabolite of vitamin D3.}, number={12}, journal={PLOS ONE}, publisher={Public Library of Science (PLoS)}, author={Kollitz, Erin M. and Zhang, Guozhu and Hawkins, Mary Beth and Whitfield, G. Kerr and Reif, David M. and Kullman, Seth W.}, editor={Zhang, ChiEditor}, year={2016}, month={Dec} }
 @article{watson_planchart_mattingly_winkler_reif_kullman_2016, title={From the Cover: Embryonic Exposure to TCDD Impacts Osteogenesis of the Axial Skeleton in Japanese medaka,Oryzias latipes}, volume={155}, ISSN={1096-6080 1096-0929}, url={http://dx.doi.org/10.1093/toxsci/kfw229}, DOI={10.1093/toxsci/kfw229}, abstractNote={Recent studies from mammalian, fish, and in vitro models have identified bone and cartilage development as sensitive targets for dioxins and other aryl hydrocarbon receptor ligands. In this study, we assess how embryonic 2,3,7,8-tetrachlorochlorodibenzo-p-dioxin (TCDD) exposure impacts axial osteogenesis in Japanese medaka (Oryzias latipes), a vertebrate model of human bone development. Embryos from inbred wild-type Orange-red Hd-dR and 3 transgenic medaka lines (twist:EGFP, osx/sp7:mCherry, col10a1:nlGFP) were exposed to 0.15 nM and 0.3 nM TCDD and reared until 20 dpf. Individuals were stained for mineralized bone and imaged using confocal microscopy to assess skeletal alterations in medial vertebrae in combination with a qualitative spatial analysis of osteoblast and osteoblast progenitor cell populations. Exposure to TCDD resulted in an overall attenuation of vertebral ossification characterized by truncated centra, and reduced neural and hemal arch lengths. Effects on mineralization were consistent with modifications in cell number and cell localization of transgene-labeled osteoblast and osteoblast progenitor cells. Endogenous expression of osteogenic regulators runt-related transcription factor 2 (runx2) and osterix (osx/sp7), and extracellular matrix genes osteopontin (spp1), collagen type I alpha I (col1), collagen type X alpha I (col10a1), and osteocalcin (bglap/osc) was significantly diminished at 20 dpf following TCDD exposure as compared with controls. Through global transcriptomic analysis more than 590 differentially expressed genes were identified and mapped to select pathological states including inflammatory disease, connective tissue disorders, and skeletal and muscular disorders. Taken together, results from this study suggest that TCDD exposure inhibits axial bone formation through dysregulation of osteoblast differentiation. This approach highlights the advantages and sensitivity of using small fish models to investigate how xenobiotic exposure may impact skeletal development.}, number={2}, journal={Toxicological Sciences}, publisher={Oxford University Press (OUP)}, author={Watson, AtLee T. D. and Planchart, Antonio and Mattingly, Carolyn J. and Winkler, Christoph and Reif, David M. and Kullman, Seth W.}, year={2016}, month={Nov}, pages={485–496} }
 @article{pow_yost_aday_kullman_2016, title={Sharing the Roles: An Assessment of Japanese Medaka Estrogen Receptors in Vitellogenin Induction}, volume={50}, ISSN={["1520-5851"]}, DOI={10.1021/acs.est.6b01968}, abstractNote={Teleost fish express at least three estrogen receptor (ER) subtypes. To date, however, the individual role of these ER subtypes in regulating expression of estrogen responsive genes remains ambiguous. Here, we investigate putative roles of three ER subtypes in Japanese medaka (Oryzias latipes), using vitellogenin (VTG) I and II as model genes. We identify specific ligand/receptor/promoter dynamics, using transient transactivation assays that incorporate luciferase reporters comprising 3kb promoter/enhancer regions of medaka VTGI and VTGII genes. Four steroidal estrogens (17β-estradiol, estrone, estriol, and 17α-estradiol) were tested in these assays. Results indicate that all three medaka ERs (mERs) are capable of initiating transactivation of both VTG I and II, with ERβ2 exhibiting greatest activity. Promoter deletion analysis suggests that ligand-specific receptor transactivation and utilization of regional-specific estrogen response elements may be associated with differential activities of each medaka ER. Further, cluster analysis of in vivo gene expression and in vitro transactivation suggests that all three ER subtypes putatively play a role in up-regulation of VTG. Results illustrate that preferential ligand/receptor/promoter interactions may have direct implications for VTG gene expression and other ER-mediated regulatory functions that are relevant to the risk assessment of estrogenic compounds.}, number={16}, journal={ENVIRONMENTAL SCIENCE & TECHNOLOGY}, author={Pow, Crystal S. D. Lee and Yost, Erin E. and Aday, D. Derek and Kullman, Seth W.}, year={2016}, month={Aug}, pages={8886–8895} }
 @misc{wilson_leblanc_kullman_crofton_schmieder_jacobs_2016, title={Where do we go from here: Challenges and the future of endocrine disrupting compound screening and testing}, url={http://dx.doi.org/10.7287/peerj.preprints.2605v1}, DOI={10.7287/peerj.preprints.2605v1}, abstractNote={Worldwide concern about the impacts of endocrine disrupting compounds on both human and environmental health has led to implementation of multiple screening and testing programs. In most cases these programs have focused on impacts to the estrogen, androgen and thyroid hormone (EAT) signaling pathways. The goal of the presentations in session five of the Society of Environmental Toxicology and Chemistry (SETAC) North America Focused Topic Meeting: Endocrine Disruption (February 4 – 6, 2014) was to discuss moving beyond EAT pathways to address current challenges and identify future approaches for the expansion of screening and testing programs. The session was chaired by Drs. Gerald A. LeBlanc and Vickie S. Wilson and included five presentations. Dr. Gerald A. LeBlanc provided insight on non-EAT endocrine targets that are known to be susceptible to endocrine disrupting compounds. Dr. Seth Kullman gave an overview of emerging technologies that hold promise for the screening of chemicals for interaction with EAT and other endocrine pathways. These were followed by two presentations on the current status and future promise of computational (Dr. Kevin Crofton) and in silico (Dr. Patricia Schmieder) approaches for screening and ranking chemicals for endocrine activity. Dr. Miriam Jacobs culminated the session with an overview of the current understanding of the role of epigenetics in endocrine regulation and approaches for evaluating chemicals for their ability to disrupt the epigenetic regulation of endocrine processes.}, publisher={PeerJ}, author={Wilson, Vickie S and LeBlanc, Gerald A and Kullman, Seth and Crofton, Kevin and Schmieder, Patricia and Jacobs, Miriam N}, year={2016}, month={Nov} }
 @article{kullman_2015, title={In Response
: Conservation versus functional diversification of nuclear receptors: An academic perspective}, volume={34}, ISSN={0730-7268}, url={http://dx.doi.org/10.1002/ETC.2832}, DOI={10.1002/ETC.2832}, abstractNote={Environmental Toxicology and ChemistryVolume 34, Issue 3 p. 463-465 ET&C Perspectives In Response: Conservation versus functional diversification of nuclear receptors: An academic perspective Seth W. Kullman, Seth W. Kullman North Carolina State University Raleigh, North Carolina, USASearch for more papers by this author Seth W. Kullman, Seth W. Kullman North Carolina State University Raleigh, North Carolina, USASearch for more papers by this author First published: 24 February 2015 https://doi.org/10.1002/etc.2832Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Volume34, Issue3March 2015Pages 463-465 RelatedInformation}, number={3}, journal={Environmental Toxicology and Chemistry}, publisher={Wiley}, author={Kullman, Seth W.}, year={2015}, month={Feb}, pages={463–465} }
 @article{kollitz_zhang_hawkins_whitfield_reif_kullman_2015, title={Molecular Cloning, Functional Characterization, and Evolutionary Analysis of Vitamin D Receptors Isolated from Basal Vertebrates}, volume={10}, ISSN={["1932-6203"]}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-84929469049&partnerID=MN8TOARS}, DOI={10.1371/journal.pone.0122853}, abstractNote={The vertebrate genome is a result of two rapid and successive rounds of whole genome duplication, referred to as 1R and 2R. Furthermore, teleost fish have undergone a third whole genome duplication (3R) specific to their lineage, resulting in the retention of multiple gene paralogs. The more recent 3R event in teleosts provides a unique opportunity to gain insight into how genes evolve through specific evolutionary processes. In this study we compare molecular activities of vitamin D receptors (VDR) from basal species that diverged at key points in vertebrate evolution in order to infer derived and ancestral VDR functions of teleost paralogs. Species include the sea lamprey (Petromyzon marinus), a 1R jawless fish; the little skate (Leucoraja erinacea), a cartilaginous fish that diverged after the 2R event; and the Senegal bichir (Polypterus senegalus), a primitive 2R ray-finned fish. Saturation binding assays and gel mobility shift assays demonstrate high affinity ligand binding and classic DNA binding characteristics of VDR has been conserved across vertebrate evolution. Concentration response curves in transient transfection assays reveal EC50 values in the low nanomolar range, however maximum transactivational efficacy varies significantly between receptor orthologs. Protein-protein interactions were investigated using co-transfection, mammalian 2-hybrid assays, and mutations of coregulator activation domains. We then combined these results with our previous study of VDR paralogs from 3R teleosts into a bioinformatics analysis. Our results suggest that 1, 25D3 acts as a partial agonist in basal species. Furthermore, our bioinformatics analysis suggests that functional differences between VDR orthologs and paralogs are influenced by differential protein interactions with essential coregulator proteins. We speculate that we may be observing a change in the pharmacodynamics relationship between VDR and 1, 25D3 throughout vertebrate evolution that may have been driven by changes in protein-protein interactions between VDR and essential coregulators.}, number={4}, journal={PLOS ONE}, author={Kollitz, Erin M. and Zhang, Guozhu and Hawkins, Mary Beth and Whitfield, G. Kerr and Reif, David M. and Kullman, Seth W.}, year={2015}, month={Apr} }
 @article{lee_kullman_yost_meyer_worley-davis_williams_reckhow_2015, title={Predicting characteristics of rainfall driven estrogen runoff and transport from swine AFO spray fields}, volume={532}, ISSN={["1879-1026"]}, DOI={10.1016/j.scitotenv.2015.06.051}, abstractNote={Animal feeding operations (AFOs) have been implicated as potentially major sources of estrogenic contaminants into the aquatic environment due to the relatively minimal treatment of waste and potential mobilization and transport of waste components from spray fields. In this study a Bayesian network (BN) model was developed to inform management decisions and better predict the transport and fate of natural steroidal estrogens from these sites. The developed BN model integrates processes of surface runoff and sediment loss with the modified universal soil loss equation (MUSLE) and the soil conservation service curve number (SCS-CN) runoff model. What-if scenario simulations of lagoon slurry wastes to the spray fields were conducted for the most abundant natural estrogen estrone (E1) observed in the system. It was found that E1 attenuated significantly after 2 months following waste slurry application in both spring and summer seasons, with the overall attenuation rate predicted to be higher in the summer compared to the spring. Using simulations of rainfall events in conjunction with waste slurry application rates, it was predicted that the magnitude of E1 runoff loss is significantly higher in the spring as compared to the summer months, primarily due to spray field crop management plans. Our what-if scenario analyses suggest that planting Bermuda grass in the spray fields is likely to reduce runoff losses of natural estrogens near the water bodies and ecosystems, as compared to planting of soybeans.}, journal={SCIENCE OF THE TOTAL ENVIRONMENT}, author={Lee, Boknam and Kullman, Sethw. and Yost, Erin E. and Meyer, Michael T. and Worley-Davis, Lynn and Williams, C. Michael and Reckhow, Kenneth H.}, year={2015}, month={Nov}, pages={571–580} }
 @article{sackett_pow_rubino_aday_cope_kullman_rice_kwak_law_2015, title={Sources of endocrine-disrupting compounds in North Carolina waterways: A geographic information systems approach}, volume={34}, ISSN={0730-7268}, url={http://dx.doi.org/10.1002/ETC.2797}, DOI={10.1002/etc.2797}, abstractNote={Abstract}, number={2}, journal={Environmental Toxicology and Chemistry}, publisher={Wiley}, author={Sackett, Dana K. and Pow, Crystal Lee and Rubino, Matthew J. and Aday, D. Derek and Cope, W. Gregory and Kullman, Seth and Rice, James A. and Kwak, Thomas J. and Law, Mac}, year={2015}, month={Jan}, pages={437–445} }
 @article{lee_kullman_yost_meyer_worley-davis_williams_reckhow_2014, title={A Bayesian Network Model for Assessing Natural Estrogen Fate and Transport in a Swine Waste Lagoon}, volume={10}, ISSN={["1551-3793"]}, DOI={10.1002/ieam.1538}, abstractNote={ABSTRACT}, number={4}, journal={INTEGRATED ENVIRONMENTAL ASSESSMENT AND MANAGEMENT}, author={Lee, Boknam and Kullman, Seth W. and Yost, Erin and Meyer, Michael T. and Worley-Davis, Lynn and Williams, C. Michael and Reckhow, Kenneth H.}, year={2014}, month={Oct}, pages={511–521} }
 @article{yost_pow_hawkins_kullman_2014, title={Bridging the Gap From Screening Assays to Estrogenic Effects in Fish: Potential Roles of Multiple Estrogen Receptor Subtypes}, volume={48}, ISSN={["1520-5851"]}, DOI={10.1021/es404093n}, abstractNote={This study seeks to delineate the ligand interactions that drive biomarker induction in fish exposed to estrogenic pollutants and provide a case study on the capacity of human (h) estrogen receptor (ER)-based in vitro screening assays to predict estrogenic effects in aquatic species. Adult male Japanese medaka (Oryzias latipes) were exposed to solutions of singular steroidal estrogens or to the estrogenic extract of an anaerobic swine waste lagoon. All exposure concentrations were calibrated to be equipotent based on the yeast estrogen screen (YES), which reports activation of hERα. These exposures elicited significantly different magnitudes of hepatic vitellogenin and choriogenin gene induction in the male medaka. Effects of the same YES-calibrated solutions in the T47D-KBluc assay, which reports activation of hERα and hERβ, generally recapitulated observations in medaka. Using competitive ligand binding assays, it was found that the magnitude of vitellogenin/choriogenin induction by different estrogenic ligands correlated positively with preferential binding affinity for medaka ERβ subtypes, which are highly expressed in male medaka liver prior to estrogen exposure. Results support emerging evidence that ERβ subtypes are critically involved in the teleost estrogenic response, with the ERα:ERβ ratio being of particular importance. Accordingly, incorporation of multiple ER subtypes into estrogen screening protocols may increase predictive value for the risk assessment of aquatic systems, including complex estrogenic mixtures.}, number={9}, journal={ENVIRONMENTAL SCIENCE & TECHNOLOGY}, author={Yost, Erin E. and Pow, Crystal Lee and Hawkins, Mary Beth and Kullman, Seth W.}, year={2014}, month={May}, pages={5211–5219} }
 @article{kollitz_hawkins_whitfield_kullmam_2014, title={Functional diversification of vitamin D receptor paralogs in teleost fish after a whole genome duplication event}, volume={155}, DOI={10.1210/en.2014-1505}, abstractNote={The diversity and success of teleost fishes (Actinopterygii) has been attributed to three successive rounds of whole-genome duplication (WGD). WGDs provide a source of raw genetic material for evolutionary forces to act upon, resulting in the divergence of genes with altered or novel functions. The retention of multiple gene pairs (paralogs) in teleosts provides a unique opportunity to study how genes diversify and evolve after a WGD. This study examines the hypothesis that vitamin D receptor (VDR) paralogs (VDRα and VDRβ) from two distantly related teleost orders have undergone functional divergence subsequent to the teleost-specific WGD. VDRα and VDRβ paralogs were cloned from the Japanese medaka (Beloniformes) and the zebrafish (Cypriniformes). Initial transactivation studies using 1α, 25-dihydroxyvitamin D3 revealed that although VDRα and VDRβ maintain similar ligand potency, the maximum efficacy of VDRβ was significantly attenuated compared with VDRα in both species. Subsequent analyses revealed that VDRα and VDRβ maintain highly similar ligand affinities; however, VDRα demonstrated preferential DNA binding compared with VDRβ. Protein-protein interactions between the VDR paralogs and essential nuclear receptor coactivators were investigated using transactivation and mammalian two-hybrid assays. Our results imply that functional differences between VDRα and VDRβ occurred early in teleost evolution because they are conserved between distantly related species. Our results further suggest that the observed differences may be associated with differential protein-protein interactions between the VDR paralogs and coactivators. We speculate that the observed functional differences are due to subtle ligand-induced conformational differences between the two paralogs, leading to divergent downstream functions.}, number={12}, journal={Endocrinology}, author={Kollitz, E. M. and Hawkins, M. B. and Whitfield, G. K. and Kullmam, Seth}, year={2014}, pages={4641–4654} }
 @article{yost_meyer_dietze_williams_worley-davis_lee_kullman_2014, title={Transport of Steroid Hormones, Phytoestrogens, and Estrogenic Activity across a Swine Lagoon/Sprayfield System}, volume={48}, ISSN={["1520-5851"]}, DOI={10.1021/es5025806}, abstractNote={The inflow, transformation, and attenuation of natural steroid hormones and phytoestrogens and estrogenic activity were assessed across the lagoon/sprayfield system of a prototypical commercial swine sow operation. Free and conjugated steroid hormones (estrogens, androgens, and progesterone) were detected in urine and feces of sows across reproductive stages, with progesterone being the most abundant steroid hormone. Excreta also contained phytoestrogens indicative of a soy-based diet, particularly, daidzein, genistein, and equol. During storage in barn pits and the anaerobic lagoon, conjugated hormones dissipated, and androgens and progesterone were attenuated. Estrone and equol persisted along the waste disposal route. Following application of lagoon slurry to agricultural soils, all analytes exhibited attenuation within 2 days. However, analytes including estrone, androstenedione, progesterone, and equol remained detectable in soil at 2 months postapplication. Estrogenic activity in the yeast estrogen screen and T47D-KBluc in vitro bioassays generally tracked well with analyte concentrations. Estrone was found to be the greatest contributor to estrogenic activity across all sample types. This investigation encompasses the most comprehensive suite of natural hormone and phytoestrogen analytes examined to date across a livestock lagoon/sprayfield and provides global insight into the fate of these analytes in this widely used waste management system.}, number={19}, journal={ENVIRONMENTAL SCIENCE & TECHNOLOGY}, author={Yost, Erin E. and Meyer, Michael T. and Dietze, Julie E. and Williams, C. Michael and Worley-Davis, Lynn and Lee, Boknam and Kullman, Seth W.}, year={2014}, month={Oct}, pages={11600–11609} }
 @book{kullman_2013, title={A Set of Scientific Issues Being Considered by the Environmental Protection Agency Regarding: Proposed Endocrine Disruptor Screening Program (EDSP) Tier 2 Ecotoxicity Tests}, number={2013-04}, institution={Environmental Protection Agency}, author={Kullman, Seth}, year={2013}, month={Jul} }
 @article{yost_meyer_dietze_meissner_worley-davis_williams_lee_kullman_2013, title={Comprehensive Assessment of Hormones, Phytoestrogens, and Estrogenic Activity in an Anaerobic Swine Waste Lagoon}, volume={47}, ISSN={["1520-5851"]}, DOI={10.1021/es4026408}, abstractNote={In this study, the distribution of steroid hormones, phytoestrogens, and estrogenic activity was thoroughly characterized within the anaerobic waste lagoon of a typical commercial swine sow operation. Three independent rounds of sampling were conducted in June 2009, April 2010, and February 2011. Thirty-seven analytes in lagoon slurry and sludge were assessed using LC/MS-MS, and yeast estrogen screen was used to determine estrogenic activity. Of the hormone analytes, steroidal estrogens were more abundant than androgens or progesterone, with estrone being the predominant estrogen species. Conjugated hormones were detected only at low levels. The isoflavone metabolite equol was by far the predominant phytoestrogen species, with daidzein, genistein, formononetin, and coumestrol present at lower levels. Phytoestrogens were often more abundant than steroidal estrogens, but contributed minimally toward total estrogenic activity. Analytes were significantly elevated in the solid phases of the lagoon; although low observed log KOC values suggest enhanced solubility in the aqueous phase, perhaps due to dissolved or colloidal organic carbon. The association with the solid phase, as well as recalcitrance of analytes to anaerobic degradation, results in a markedly elevated load of analytes and estrogenic activity within lagoon sludge. Overall, findings emphasize the importance of adsorption and transformation processes in governing the fate of these compounds in lagoon waste, which is ultimately used for broadcast application as a fertilizer.}, number={23}, journal={ENVIRONMENTAL SCIENCE & TECHNOLOGY}, author={Yost, Erin E. and Meyer, Michael T. and Dietze, Julie E. and Meissner, Benjamin M. and Worley-Davis, Lynn and Williams, C. Michael and Lee, Boknam and Kullman, Seth W.}, year={2013}, month={Dec}, pages={13781–13790} }
 @article{wettere_kullmam_hinton_law_2013, title={Immunohistochemical characterization of the hepatic progenitor cell compartment in medaka (oryzias latipes) following hepatic injury}, volume={149}, DOI={10.1016/j.jcpa.2013.03.008}, abstractNote={Laboratory fish species are used increasingly in biomedical research and are considered robust models for the study of regenerative processes. Studies investigating the response of the fish liver to injury have demonstrated the presence of a ductular reaction and oval-like cells in injured and regenerating liver. To date, however, it is unclear if this cell population is the piscine equivalent of oval cells (OCs) or intermediate hepatobiliary cells (IHBCs) identified in rodents and man, respectively. The present study defines the process of OC differentiation in fish liver using histopathology, immunohistochemistry and transmission electron microscopy. To generate OC proliferation in Japanese medaka (Oryzias latipes), hepatic injury was induced by exposure of adult fish to either microcystin LR or dimethylnitrosamine. A transgenic strain of medaka expressing a red fluorescent protein (RFP) exclusively in hepatocytes was used. The morphological response to injury was characterized by a ductular reaction comprised of cytokeratin (CK) AE1/AE3+ OCs progressing to IHBCs variably positive for CK and RFP and finally mature RFP+ hepatocytes and CK+ cholangiocytes. These observations support a bipotential differentiation pathway of fish OCs towards hepatocytes and cholangiocytes. Ultrastructural morphology confirmed the presence of OCs and differentiation towards hepatocytes. These results demonstrated clear similarities between patterns of reaction to injury in fish and mammalian livers. They also confirm the presence of, and support the putative bipotential lineage capabilities of, the fish OC.}, number={4}, journal={Journal of Comparative Pathology}, author={Wettere, A. J. Van and Kullmam, Seth and Hinton, D. E. and Law, J. M.}, year={2013}, pages={434–445} }
 @article{van wettere_law_hinton_kullman_2013, title={Phenotypic Characterization of Transgenic Japanese Medaka (Oryzias latipes) That Express a Red Fluorescent Protein in Hepatocytes}, volume={42}, ISSN={0192-6233 1533-1601}, url={http://dx.doi.org/10.1177/0192623313499643}, DOI={10.1177/0192623313499643}, abstractNote={ Transgenic organisms that express fluorescent proteins are used frequently for in vivo visualization of proteins and cells. The phenotype of a transgenic medaka ( Oryzias latipes) strain that expresses a red fluorescent protein (RFP) in hepatocytes was characterized using light and fluorescence microscopy, immunohistochemistry, and transmission electron microscopy (TEM). Expression of RFP was first detected by confocal fluorescence microscopy in the location of the liver bud of live medaka embryos at 60 hr postfertilization (developmental stage 27). Subsequently, RFP signal was observed exclusively in hepatocytes throughout life using fluorescence microscopy in live fish and immunohistochemistry in formalin-fixed, paraffin-embedded liver sections. As the fish aged, prominent intracytoplasmic eosinophilic inclusions immunoreactive for RFP were observed by light microscopy and were correlated with membrane-bound electron dense inclusions on TEM. These results define the onset and location of RFP expression in the Tg(zf.L-fabp:DsRed) medaka and characterize a histologic phenotype that results from RFP expression in hepatocytes. }, number={3}, journal={Toxicologic Pathology}, publisher={SAGE Publications}, author={Van Wettere, Arnaud J. and Law, J. Mac and Hinton, David E. and Kullman, Seth W.}, year={2013}, month={Aug}, pages={616–621} }
 @article{miller_clark_hinton_whitehead_martin_kwok_kullman_2012, title={Anchoring Ethinylestradiol Induced Gene Expression Changes with Testicular Morphology and Reproductive Function in the Medaka}, volume={7}, ISSN={["1932-6203"]}, DOI={10.1371/journal.pone.0052479}, abstractNote={Environmental estrogens are ubiquitous in the environment and can cause detrimental effects on male reproduction. In fish, a multitude of effects from environmental estrogens have been observed including altered courting behavior and fertility, sex reversal, and gonadal histopathology. However, few studies in fish assess the impacts of estrogenic exposure on a physiological endpoint, such as reproduction, as well as the associated morphologic response and underlying global gene expression changes. This study assessed the implications of a 14 day sub-chronic exposure of ethinylestradiol (EE2; 1.0 or 10.0 µg/L EE2) on male medaka fertility, testicular histology and testicular gene expression. The findings demonstrate that a 14 day exposure to EE2 induced impaired male reproductive capacity and time- and dose-dependent alterations in testicular morphology and gene expression. The average fertilization rate/day following the exposure for control, 1.0 and 10.0 µg/L EE2 was 91.3% (±4.4), 62.8% (±8.3) and 28.8% (±5.8), respectively. The testicular morphologic alterations included increased germ cell apoptosis, decreased germinal epithelium and thickening of the interstitium. These changes were highly associated with testicular gene expression changes using a medaka-specific microarray. A pathway analysis of the differentially expressed genes emphasized genes and pathways associated with apoptosis, cell cycle and proliferation, collagen production/extracellular matrix organization, hormone signaling, male reproduction and protein ubiquitination among others. These findings highlight the importance of anchoring global gonadal gene expression changes with morphology and ultimately with tissue/organ function.}, number={12}, journal={PLOS ONE}, author={Miller, Hilary D. and Clark, Bryan W. and Hinton, David E. and Whitehead, Andrew and Martin, Stan and Kwok, Kevin W. and Kullman, Seth W.}, year={2012}, month={Dec} }
 @article{van wettere_law_hinton_kullman_2012, title={Anchoring hepatic gene expression with development of fibrosis and neoplasia in a toxicant-induced fish model of liver injury}, volume={41}, ISSN={["1533-1601"]}, url={http://tpx.}, DOI={10.1177/0192623312464308}, abstractNote={ Fish have been used as laboratory models to study hepatic development and carcinogenesis but not for pathogenesis of hepatic fibrosis. In this study, a dimethylnitrosamine-induced fish model of hepatic injury was developed in Japanese medaka ( Oryzias latipes) and gene expression was anchored with the development of hepatic fibrosis and neoplasia. Exposed livers exhibited mild hepatocellular degenerative changes 2 weeks’ postexposure. Within 6 weeks, hepatic fibrosis/cirrhosis was evident with development of neoplasia by 10 weeks. Stellate cell activation and development of fibrosis was associated with upregulation of transforming growth factor beta 1 ( tgfb1), tgfb receptor 2, mothers against decapentaplegic homolog 3 ( smad3a), smad3b, beta-catenin ( ctnnb1), myc, matrix metalloproteinase ( mmp2), mmp14a, mmp14b, tissue inhibitors of metalloproteinase ( timp) 2a, timp2b, timp3, collagen type I alpha 1a ( col1a1a), and col1a1b and a less pronounced increase in mmp13 and col4a1 expression. Tgfb receptor I expression was unchanged. Immunohistochemistry suggested that biliary epithelial cells and stellate cells were the main producers of TGF-β1. This study identified a group of candidate genes likely to be involved in the development of hepatic fibrosis and demonstrated that the TGF-β pathway likely plays a major role in the pathogenesis. These results support the medaka as a viable fish model of hepatic fibrosis. }, number={5}, journal={Toxicologic Pathology}, author={Van Wettere, A.J. and Law, J.M. and Hinton, D.E. and Kullman, S.W.}, year={2012}, pages={1–17} }
 @article{law_redelings_kullman_2012, title={Comparative Genomics of Duplicate gamma-Glutamyl Transferase Genes in Teleosts: Medaka (Oryzias latipes), Stickleback (Gasterosteus aculeatus), Green Spotted Pufferfish (Tetraodon nigroviridis), Fugu (Takifugu rubripes), and Zebrafish (Danio rerio)}, volume={318B}, ISSN={["1552-5015"]}, DOI={10.1002/jez.b.21439}, abstractNote={Abstract}, number={1}, journal={JOURNAL OF EXPERIMENTAL ZOOLOGY PART B-MOLECULAR AND DEVELOPMENTAL EVOLUTION}, author={Law, Sheran Hiu Wan and Redelings, Benjamin David and Kullman, Seth William}, year={2012}, month={Jan}, pages={35–49} }
 @book{detailed review paper on the state of the science on novel in vitro and in vivo screening and testing methods and endpoints for evaluating endocrine disruptors_2012, ISBN={9789264221352}, ISSN={2077-7876}, url={http://dx.doi.org/10.1787/9789264221352-en}, DOI={10.1787/9789264221352-en}, abstractNote={Increasing incidents of disorders such as obesity/diabetes/metabolic syndrome, reproductive dysfunction, and neuro-developmental abnormalities in some human populations have raised concern that disruption of key endocrine-signaling pathways by exposure to environmental chemicals may be involved.This Detailed Review Paper describes some endocrine pathways that have been shown to be susceptible to environmental endocrine disruption and whose disruption could contribute to increasing incidents of some disorders in humans and wildlife populations.Assays and endpoints are described that could be used in new or existing Organization for Economic Cooperative Development (OECD) Test Guidelines for evaluating chemicals for endocrine-disrupting activity.Endocrine pathways evaluated were the hypothalamus:pituitary:adrenocortical (HPA) axis, the hypothalamus:pituitary:gonad (HPG) axis, the somatotropic axis, the retinoid signaling pathway, the hypothalamus:pituitary:thyroid (HPT) axis, the vitamin D signaling pathway, and the peroxisome proliferator-activated receptor (PPAR) signaling pathway.In addition, the potential role of chemical-induced epigenetic modifications to endocrine signaling pathways, during sensitive windows of exposure, was evaluated as a mechanism of endocrine disruption, along with the examination of potential methods for assessing such disruption.This section is provided as an annex to the document (Annex 1).Potential targets of disruption along putative adverse outcome pathways associated with the signaling pathways were identified, along with assays that show promise in evaluating the target in a screening and testing program.Disruption of the HPA or retinoid X receptor signaling pathways could contribute to disorders of emerging concern, and adverse outcome pathways are well defined.However, assays for the assessment of disruption of these pathways are less well developed, and in some cases, are not specific to the pathway.Several new assays were described for the detection of disruption of the HPT axis.These assays may complement assays in the existing Test Guidelines and strengthen the adverse outcome pathway lineage.Assays for the detection of vitamin D signaling disruption and novel aspects of the HPG axis (membrane receptor signaling, progestin signaling) require further development and refinement prior to consideration for incorporation into Test Guidelines.Disruption of the somatotropic axis is likely to occur through disruption of other signaling pathways that cross-talk with the somatotropic axis.Disruption of the somatotropic axis may thus provide a more holistic view of the general integrity of the endocrine system.PPARs are involved in lipid and glucose homeostasis, inflammation, and aspects of development.The adverse outcome pathway for PPARγ is well established.Assays used to assess disruption of PPAR signaling are well developed, and many are suitable for incorporation into existing OECD Test Guidelines.In conclusion, OECD Test Guidelines could be modified to include new assays or the incorporation of novel endpoints into existing assays that would expand the repertoire of endocrine signaling pathways included in the screening and testing regimen.* See section 1.2 1 Some assays may also provide some evidence of adverse effects. 2 Effects can be sensitive to more than one mechanism and may be due to non-endocrine mechanisms.* See section 1.2 1 Some assays may also provide some evidence of adverse effects. 2 Effects can be sensitive to more than one mechanism and may be due to non-endocrine mechanisms.* See section 1.2 1 Some assays may also provide some evidence of adverse effects. 2 Effects can be sensitive to more than one mechanism and may be due to non-endocrine mechanisms.* See section 1.2 1 Some assays may also provide some evidence of adverse effects. 2 Effects can be sensitive to more than one mechanism and may be due to non-endocrine mechanisms.* See section 1.2 1 Some assays may also provide some evidence of adverse effects. 2 Effects can be sensitive to more than one mechanism and may be due to non-endocrine mechanisms.}, journal={OECD Series on Testing and Assessment}, publisher={OECD}, year={2012}, month={Aug} }
 @article{dong_hinton_kullman_2012, title={TCDD Disrupts Hypural Skeletogenesis during Medaka Embryonic Development}, volume={125}, ISSN={["1096-6080"]}, DOI={10.1093/toxsci/kfr284}, abstractNote={Abstract}, number={1}, journal={TOXICOLOGICAL SCIENCES}, author={Dong, Wu and Hinton, David E. and Kullman, Seth W.}, year={2012}, month={Jan}, pages={91–104} }
 @article{krasowski_ai_hagey_kollitz_kullman_reschly_ekins_2011, title={The evolution of farnesoid X, vitamin D, and pregnane X receptors: insights from the green-spotted pufferfish (Tetraodon nigriviridis) and other non-mammalian species}, volume={12}, ISSN={["1471-2091"]}, DOI={10.1186/1471-2091-12-5}, abstractNote={The farnesoid X receptor (FXR), pregnane X receptor (PXR), and vitamin D receptor (VDR) are three closely related nuclear hormone receptors in the NR1H and 1I subfamilies that share the property of being activated by bile salts. Bile salts vary significantly in structure across vertebrate species, suggesting that receptors binding these molecules may show adaptive evolutionary changes in response. We have previously shown that FXRs from the sea lamprey (Petromyzon marinus) and zebrafish (Danio rerio) are activated by planar bile alcohols found in these two species. In this report, we characterize FXR, PXR, and VDR from the green-spotted pufferfish (Tetraodon nigriviridis), an actinopterygian fish that unlike the zebrafish has a bile salt profile similar to humans. We utilize homology modelling, docking, and pharmacophore studies to understand the structural features of the Tetraodon receptors.Tetraodon FXR has a ligand selectivity profile very similar to human FXR, with strong activation by the synthetic ligand GW4064 and by the primary bile acid chenodeoxycholic acid. Homology modelling and docking studies suggest a ligand-binding pocket architecture more similar to human and rat FXRs than to lamprey or zebrafish FXRs. Tetraodon PXR was activated by a variety of bile acids and steroids, although not by the larger synthetic ligands that activate human PXR such as rifampicin. Homology modelling predicts a larger ligand-binding cavity than zebrafish PXR. We also demonstrate that VDRs from the pufferfish and Japanese medaka were activated by small secondary bile acids such as lithocholic acid, whereas the African clawed frog VDR was not.Our studies provide further evidence of the relationship between both FXR, PXR, and VDR ligand selectivity and cross-species variation in bile salt profiles. Zebrafish and green-spotted pufferfish provide a clear contrast in having markedly different primary bile salt profiles (planar bile alcohols for zebrafish and sterically bent bile acids for the pufferfish) and receptor selectivity that matches these differences in endogenous ligands. Our observations to date present an integrated picture of the co-evolution of bile salt structure and changes in the binding pockets of three nuclear hormone receptors across the species studied.}, journal={BMC BIOCHEMISTRY}, author={Krasowski, Matthew D. and Ai, Ni and Hagey, Lee R. and Kollitz, Erin M. and Kullman, Seth W. and Reschly, Erica J. and Ekins, Sean}, year={2011}, month={Feb} }
 @article{howarth_law_law_mondon_kullman_hinton_2010, title={Exposure to the synthetic FXR agonist GW4064 causes alterations in gene expression and sublethal hepatotoxicity in eleutheroembryo medaka (Oryzias latipes)}, volume={243}, ISSN={["1096-0333"]}, DOI={10.1016/j.taap.2009.11.022}, abstractNote={The small freshwater teleost, medaka (Oryzias latipes), has a history of usage in studies of chronic toxicity of liver and biliary system. Recent progress with this model has focused on defining the medaka hepatobiliary system. Here we investigate critical liver function and toxicity by examining the in vivo role and function of the farnesoid X receptor alpha (FXRα, NR1H4), a member of the nuclear receptor superfamily that plays an essential role in the regulation of bile acid homeostasis. Quantitative mRNA analysis of medaka FXRα demonstrates differential expression of two FXRα isoforms designated Fxrα1 and Fxrα2, in both free swimming medaka embryos with remaining yolk (eleutheroembryos, EEs) and adults. Activation of medaka Fxrα in vivo with GW4064 (a strong FXRα agonist) resulted in modification of gene expression for defined FXRα gene targets including the bile salt export protein, small heterodimer partner, and cytochrome P450 7A1. Histological examination of medaka liver subsequent to GW4064 exposure demonstrated significant lipid accumulation, cellular and organelle alterations in both hepatocytes and biliary epithelial cells of the liver. This report of hepatobiliary injury following GW4064 exposure extends previous investigations of the intrahepatic biliary system in medaka, reveals sensitivity to toxicant exposure, and illustrates the need for added resolution in detection and interpretation of toxic responses in this vertebrate.}, number={1}, journal={TOXICOLOGY AND APPLIED PHARMACOLOGY}, author={Howarth, Deanna L. and Law, Sheran H. W. and Law, J. McHugh and Mondon, J. A. and Kullman, Seth W. and Hinton, David E.}, year={2010}, month={Feb}, pages={111–121} }
 @inbook{kullman_mattingly_meyer_whitehead_2010, place={Hoboken NJ}, edition={4th}, title={Perspectives on Informatics in Toxicology, in A Textbook of Modern Toxicology}, booktitle={A Textbook of Modern Toxicology}, publisher={John Wiley and Sons}, author={Kullman, S.W. and Mattingly, C.J. and Meyer, J.N. and Whitehead, A.}, editor={Hodgson, ErnestEditor}, year={2010}, pages={593–605} }
 @article{dong_matsumura_kullman_2010, title={TCDD Induced Pericardial Edema and Relative COX-2 Expression in Medaka (Oryzias Latipes) Embryos}, volume={118}, ISSN={["1096-0929"]}, DOI={10.1093/toxsci/kfq254}, abstractNote={Exposure to dioxin and other aryl hydrocarbon receptor (AhR) ligands results in multiple, specific developmental cardiovascular phenotypes including pericardial edema and circulatory failure in small aquarium fish models. Although phenotypes are well described, mechanistic underpinnings for such toxicities remain elusive. Here we suggest that AhR activation results in stimulation of inflammation and "eicosanoid" pathways, which contribute to the observed developmental, cardiovascular phenotypes. We demonstrate that medaka embryos exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (0.05-1 ppb) during early development result in a dose-related increase in the prevalence of pericardial edema and that this phenotype correlates with an increase in cyclooxygenase-2 (COX-2) gene expression. Those individuals exhibiting the edema phenotype had significantly greater COX-2 mRNA than their nonedematous cohort. Selective pharmacological inhibition of COX-2, with NS-398, and genetic knock down of COX-2 with a translation initiation morpholino significantly attenuated prevalence and severity of edema phenotype. Subsequently, exposures of medaka embryos to arachidonic acid (AA) resulted in recapitulation of the pericardial edema phenotype and significantly increased COX-2 expression only in those individuals exhibiting the edema phenotype compared with their nonedematous cohort. AA exposure does not result in significant induction of cytochrome P450 1A expression, suggesting that pericardial edema can be induced independent of AhR/aryl hydrocarbon receptor nuclear translocator/dioxin response element interactions. Results from this study demonstrate that developmental exposure to TCDD results in an induction of inflammatory mediators including COX-2, which contribute to the onset, and progression of heart dysmorphogenesis in the medaka model.}, number={1}, journal={TOXICOLOGICAL SCIENCES}, author={Dong, Wu and Matsumura, Fumio and Kullman, Seth W.}, year={2010}, month={Nov}, pages={213–223} }
 @article{howarth_hagey_law_ai_krasowski_ekins_moore_kollitz_hinton_kullmam_2010, title={Two farnesoid X receptor alpha isoforms in Japanese medaka (Oryzias latipes) are differentially activated in vitro}, volume={98}, DOI={10.1016/j.aquatox.2010.02.020}, abstractNote={The nuclear receptor farnesoid X receptor alpha (FXRα, NR1H4) is activated by bile acids in multiple species including mouse, rat, and human and in this study we have identified two isoforms of Fxrα in Japanese medaka (Oryzias latipes), a small freshwater teleost. Both isoforms share a high amino acid sequence identity to mammalian FXRα (∼70% in the ligand-binding domain). Fxrα1 and Fxrα2 differ within the AF1 domain due to alternative splicing at the fourth intron-exon boundary. This process results in Fxrα1 having an extended N-terminus compared to Fxrα2. A Gal4DBD-FxrαLBD fusion construct was activated by chenodeoxycholic, cholic, deoxycholic and lithocholic acids, and the synthetic agonist GW4064 in transient transactivation assays. Activation of the Gal4DBD-FxrαLBD fusion construct was enhanced by addition of PGC-1α, as demonstrated through titration assays. Surprisingly, when the full-length versions of the two Fxrα isoforms were compared in transient transfection assays, Fxrα2 was activated by C24 bile acids and GW4064, while Fxrα1 was not significantly activated by any of the compounds tested. Since the only significant difference between the full-length constructs was sequence in the AF1 domain, these experiments highlight a key functional region in the Fxrα AF1 domain. Furthermore, mammalian two-hybrid studies demonstrated the ability of Fxrα2, but not Fxrα1, to interact with PGC-1α and SRC-1, and supported our results from the transient transfection reporter gene activation assays. These data demonstrate that both mammalian and teleost FXR (Fxrα2 isoform) are activated by primary and secondary bile acids.}, number={3}, journal={Aquatic Toxicology (Amsterdam, Netherlands)}, author={Howarth, D. L. and Hagey, L. R. and Law, S. H. W. and Ai, N. and Krasowski, M. D. and Ekins, S. and Moore, J. T. and Kollitz, E. M. and Hinton, D. E. and Kullmam, Seth}, year={2010}, pages={245–255} }
 @article{hinton_hardman_kullman_law_schmale_walter_winn_yoder_2009, title={Aquatic animal models of human disease: Selected papers and recommendations from the 4th Conference}, volume={149}, ISSN={1532-0456}, url={http://dx.doi.org/10.1016/j.cbpc.2008.12.006}, DOI={10.1016/j.cbpc.2008.12.006}, abstractNote={Large volumes of oil sands process-affected water (OSPW) are produced by the surface-mining oil sands industry in Alberta. Both laboratory and field studies have demonstrated that the exposure to OSPW leads to many physiological changes in a variety of organisms. Adverse effects include compromised immunological function, developmental delays, impaired reproduction, disrupted endocrine system, and higher prevalence of tissue-specific pathological manifestations. The composition of OSPW varies with several factors such as ore sources, mining process, and tailings management practices. Differences in water characteristics have confounded interpretation or comparison of OSPW toxicity across studies. Research on individual fractions extracted from OSPW has helped identify some target pollutants. Naphthenic acids (NAs) are considered as the major toxic components in OSPW, exhibiting toxic effects through multiple modes of action including narcosis and endocrine disruption. Other pollutants, like polycyclic aromatic hydrocarbons (PAHs), metals, and ions may also contribute to the overall OSPW toxicity. Studies have been conducted on OSPW as a whole complex effluent mixture, with consideration of the presence of unidentified components, and the interactions (potential synergistic or antagonistic reactions) among chemicals. This review summarizes the toxicological data derived from in vitro and in vivo exposure studies using different OSPW types, and different taxa of organisms. In general, toxicity of OSPW was found to be dependent on the OSPW type and concentration, duration of exposures (acute versus sub chronic), and organism studied.}, number={2}, journal={Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology}, publisher={Elsevier BV}, author={Hinton, David E. and Hardman, Ron C. and Kullman, Seth W. and Law, Jerry M. and Schmale, Michael C. and Walter, Ronald B. and Winn, Richard N. and Yoder, Jeffrey A.}, year={2009}, month={Mar}, pages={121–128} }
 @article{padilla_cowden_hinton_yuen_law_kullman_johnson_hardman_flynn_au_2009, title={Use of Medaka in Toxicity Testing}, volume={39}, ISSN={1934-9254 1934-9262}, url={http://dx.doi.org/10.1002/0471140856.tx0110s39}, DOI={10.1002/0471140856.tx0110s39}, abstractNote={Abstract}, number={1}, journal={Current Protocols in Toxicology}, publisher={Wiley}, author={Padilla, Stephanie and Cowden, John and Hinton, David E. and Yuen, Bonny and Law, Sheran and Kullman, Seth W. and Johnson, Rodney and Hardman, Ronald C. and Flynn, Kevin and Au, Doris W.T.}, year={2009}, month={Feb} }
 @inbook{schlenk_celander_gallagher_george_james_kullman_van den hurk_willett_2008, place={Boca Raton, Fl}, title={Biotransformation in Fishes}, url={http://dx.doi.org/10.1201/9780203647295.ch4}, DOI={10.1201/9780203647295.ch4}, booktitle={The Toxicology of Fishes}, publisher={CRC Press}, author={Schlenk, Daniel and Celander, Malin and Gallagher, Evan and George, Stephen and James, Margaret and Kullman, Seth and van den Hurk, Peter and Willett, Kristie}, year={2008}, month={Feb}, pages={153–234} }
 @article{carney_erwin_hardman_yuen_volz_hinton_kullman_2008, title={Differential developmental toxicity of naphthoic acid isomers in medaka (Oryzias latipes) embryos}, volume={57}, ISSN={["1879-3363"]}, DOI={10.1016/j.marpolbul.2008.02.036}, abstractNote={Polycyclic aromatic hydrocarbons (PAHs) are widespread persistent pollutants that readily undergo biotic and abiotic conversion to numerous transformation products in rivers, lakes and estuarine sediments. Here we characterize the developmental toxicity of four PAH transformation products each structural isomers of hydroxynaphthoic acid: 1H2NA, 2H1NA, 2H3NA, and 6H2NA. Medaka fish (Oryzias latipes) embryos and eleutheroembryos were used to determine toxicity. A 96-well micro-plate format was used to establish a robust, statistically significant platform for assessment of early life stages. Individual naphthoic acid isomers demonstrated a rank order of toxicity with 1H2NA>2H1NA>2H3NA>6H2NA being more toxic. Abnormalities of circulatory system were most pronounced including pericardial edema and tube heart. To determine if HNA isomers were AhR ligands, spatial-temporal expression and activity of CYP1A was measured via in vivo EROD assessments. qPCR measurement of CYP1A induction proved different between isomers dosed at respective concentrations affecting 50% of exposed individuals (EC50s). In vitro, all ANH isomers transactivated mouse AhR using a medaka CYP1A promoter specific reporter assay. Circulatory abnormalities followed P450 induction and response was consistent with PAH toxicity. A 96-well micro-plates proved suitable as exposure chambers and provided statistically sound evaluations as well as efficient toxicity screens. Our results demonstrate the use of medaka embryos for toxicity analysis thereby achieving REACH objectives for the reduction of adult animal testing in toxicity evaluations.}, number={6-12}, journal={MARINE POLLUTION BULLETIN}, author={Carney, Michael W. and Erwin, Kyle and Hardman, Ron and Yuen, Bonny and Volz, David C. and Hinton, David E. and Kullman, Seth W.}, year={2008}, pages={255–266} }
 @book{kullman_2008, title={Endocrine Disruptor Screening Program (EDSP) Proposed Tier 1 Screening Battery - Federal Insecticide, Fungicide, and Rodenticide Act Scientific Advisory Panel}, institution={Environmental Protection Agency}, author={Kullman, S.}, year={2008}, month={Mar} }
 @inbook{hinton_segner_au_kullman_hardman_2008, place={Boca Raton, FL}, title={Liver Toxicity}, url={http://dx.doi.org/10.1201/9780203647295.ch7}, DOI={10.1201/9780203647295.ch7}, booktitle={The Toxicology of Fishes}, publisher={CRC Press}, author={Hinton, David and Segner, Helmut and Au, Doris and Kullman, Seth and Hardman, Ronald}, year={2008}, month={Feb}, pages={327–400} }
 @article{hardman_kullman_yuen_hinton_2008, title={Non invasive high resolution in vivo imaging of α-naphthylisothiocyanate (ANIT) induced hepatobiliary toxicity in STII medaka}, volume={86}, ISSN={0166-445X}, url={http://dx.doi.org/10.1016/j.aquatox.2007.09.014}, DOI={10.1016/j.aquatox.2007.09.014}, abstractNote={A novel transparent stock of medaka (Oryzias latipes; STII), homozygous recessive for all four pigments (iridophores, xanthophores, leucophores, melanophores), permits transcutaneous, high resolution (<1 μm) imaging of internal organs and tissues in living individuals. We applied this model to in vivo investigation of α -naphthylisothiocyanate (ANIT) induced hepatobiliary toxicity. Distinct phenotypic responses to ANIT involving all aspects of intrahepatic biliary passageways (IHBPs), particularly bile preductular epithelial cells (BPDECs), associated with transitional passageways between canaliculi and bile ductules, were observed. Alterations included: attenuation/dilation of bile canaliculi, bile preductular lesions, hydropic vacuolation of hepatocytes and BPDECs, mild BPDEC hypertrophy, and biliary epithelial cell (BEC) hyperplasia. Ex vivo histological, immunohistochemical, and ultrastructural studies were employed to aid in interpretation of, and verify, in vivo findings. 3D reconstructions from in vivo investigations provided quantitative morphometric and volumetric evaluation of ANIT exposed and untreated livers. The findings presented show for the first time in vivo evaluation of toxicity in the STII medaka hepatobiliary system, and, in conjunction with prior in vivo work characterizing normalcy, advance our comparative understanding of this lower vertebrate hepatobiliary system and its response to toxic insult.}, number={1}, journal={Aquatic Toxicology}, publisher={Elsevier BV}, author={Hardman, Ron and Kullman, Seth and Yuen, Bonny and Hinton, David E.}, year={2008}, month={Jan}, pages={20–37} }
 @article{hardman_kullman_hinton_2008, title={Non invasive in vivo investigation of hepatobiliary structure and function in STII medaka (Oryzias latipes): methodology and applications}, volume={7}, ISSN={1476-5926}, url={http://dx.doi.org/10.1186/1476-5926-7-7}, DOI={10.1186/1476-5926-7-7}, abstractNote={A novel transparent stock of medaka (Oryzias latipes; STII), recessive for all pigments found in chromatophores, permits transcutaneous imaging of internal organs and tissues in living individuals. Findings presented describe the development of methodologies for non invasive in vivo investigation in STII medaka, and the successful application of these methodologies to in vivo study of hepatobiliary structure, function, and xenobiotic response, in both 2 and 3 dimensions.Using brightfield, and widefield and confocal fluorescence microscopy, coupled with the in vivo application of fluorescent probes, structural and functional features of the hepatobiliary system, and xenobiotic induced toxicity, were imaged at the cellular level, with high resolution (< 1 microm), in living individuals. The findings presented demonstrate; (1) phenotypic response to xenobiotic exposure can be investigated/imaged in vivo with high resolution (< 1 microm), (2) hepatobiliary transport of solutes from blood to bile can be qualitatively and quantitatively studied/imaged in vivo, (3) hepatobiliary architecture in this lower vertebrate liver can be studied in 3 dimensions, and (4) non invasive in vivo imaging/description of hepatobiliary development in this model can be investigated.The non-invasive in vivo methodologies described are a unique means by which to investigate biological structure, function and xenobiotic response with high resolution in STII medaka. In vivo methodologies also provide the future opportunity to integrate molecular mechanisms (e.g., genomic, proteomic) of disease and toxicity with phenotypic changes at the cellular and system levels of biological organization. While our focus has been the hepatobiliary system, other organ systems are equally amenable to in vivo study, and we consider the potential for discovery, within the context of in vivo investigation in STII medaka, as significant.}, number={1}, journal={Comparative Hepatology}, publisher={Springer Science and Business Media LLC}, author={Hardman, Ron C and Kullman, Seth W and Hinton, David E}, year={2008}, month={Oct} }
 @article{howarth_law_barnes_hall_hinton_moore_maglich_moore_kullman_2008, title={Paralogous vitamin D receptors in teleosts: Transition of nuclear receptor function}, volume={149}, ISSN={["1945-7170"]}, DOI={10.1210/en.2007-1256}, abstractNote={The availability of multiple teleost (bony fish) genomes is providing unprecedented opportunities to understand the diversity and function of gene duplication events using comparative genomics. Here we describe the cloning and functional characterization of two novel vitamin D receptor (VDR) paralogs from the freshwater teleost medaka (Oryzias latipes). VDR sequences were identified through mining of the medaka genome database in which gene organization and structure was determined. Two distinct VDR genes were identified in the medaka genome and mapped to defined loci. Each VDR sequence exhibits unique intronic organization and dissimilar 5′ untranslated regions, suggesting they are not isoforms of the same gene locus. Phylogenetic comparison with additional teleosts and mammalian VDR sequences illustrate that two distinct clusters are formed separating aquatic and terrestrial species. Nested within the teleost cluster are two separate clades for VDRα and VDRβ. The topology of teleost VDR sequences is consistent with the notion of paralogous genes arising from a whole genome duplication event prior to teleost radiation. Functional characterization was conducted through the development of VDR expression vectors including Gal4 chimeras containing the yeast Gal4 DNA binding domain fused to the medaka VDR ligand binding domain and full-length protein. The common VDR ligand 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D3] resulted in significant transactivation activity with both the Gal4 and full-length constructs of medaka (m) VDRβ. Comparatively, transactivation of mVDRα with 1α,25(OH)2D3 was highly attenuated, suggesting a functional divergence between these two nuclear receptor paralogs. We additionally demonstrate through coactivator studies that mVDRα is still functional; however, it exhibits a different sensitivity to 1α,25(OH)2D3, compared with VDRβ. These results suggest that in mVDRα and VDRβ have undergone a functional divergence through a process of sub- and/or neofunctionalization of VDR nuclear receptor gene pairs.}, number={5}, journal={ENDOCRINOLOGY}, author={Howarth, Deanna L. and Law, Sheran H. W. and Barnes, Benjamin and Hall, Julie M. and Hinton, David E. and Moore, Linda and Maglich, Jodi M. and Moore, John T. and Kullman, Seth W.}, year={2008}, month={May}, pages={2411–2422} }
 @article{volz_kullman_howarth_hardman_hinton_2008, title={Protective response of the ah receptor to ANIT-Induced biliary epithelial cell toxicity in see-through medaka}, volume={102}, ISSN={["1096-0929"]}, DOI={10.1093/toxsci/kfm308}, abstractNote={The adaptive role of the aryl hydrocarbon receptor (Ah receptor or AHR) in protecting against disease-related conditions remains unclear in nonmammalian models, particularly teleosts. Therefore, this study focused on the potential role of AHR in response to biliary epithelial cell toxicity and hepatobiliary alteration in medaka. See-through medaka (STII strain) were exposed for 96 h using the biliary toxicant alpha-naphthylisothiocyanate (ANIT) as a reagent, and fish were evaluated daily using histological and ultrastructural analysis, and by imaging directly through the body wall of living fish. Brightfield and transmission electron microscopy showed that a single ANIT dose (40 mg/kg) specifically induced swelling and apoptosis of bile preductular epithelial cells (BPDECs) as early as 6 h after initial exposure. Following ANIT-induced BPDEC toxicity, in vivo imaging of STII medaka showed significant gallbladder discoloration from 48-72 h. Collectively, these pathologic data suggested that ANIT exposure resulted in acute hepatobiliary changes, lasting < 96 h following initial exposure. We then tested the potential role of AHR in response to ANIT-induced hepatobiliary alteration. Overall, we demonstrated that (1) transient AHR activation and cytochrome P450 1A (CYP1A) induction in livers occurred during ANIT-induced hepatobiliary impairment, (2) pretreatment with an AHR agonist partially protected against acute hepatobiliary alteration, and (3) using a luciferase-based reporter assay, the bile pigment bilirubin weakly activated mouse AHR and binding to medaka-specific CYP1A promoter, resulting in AHR element-driven transcription. Given that bile acids and pigments are present in mammalian and fish liver, these studies collectively suggest that bile-induced AHR activation may be conserved between teleosts and rodents.}, number={2}, journal={TOXICOLOGICAL SCIENCES}, author={Volz, David C. and Kullman, Seth W. and Howarth, Deanna L. and Hardman, Ron C. and Hinton, David E.}, year={2008}, month={Apr}, pages={262–277} }
 @article{hardman_volz_kullman_hinton_2007, title={An in vivo Look at Vertebrate Liver Architecture: Three-Dimensional Reconstructions from Medaka (Oryzias latipes)}, volume={290}, ISSN={1932-8486 1932-8494}, url={http://dx.doi.org/10.1002/ar.20524}, DOI={10.1002/ar.20524}, abstractNote={Abstract}, number={7}, journal={The Anatomical Record: Advances in Integrative Anatomy and Evolutionary Biology}, publisher={Wiley}, author={Hardman, Ron C. and Volz, Dave C. and Kullman, Seth W. and Hinton, David E.}, year={2007}, pages={770–782} }
 @article{hardman_kullman_hinton_2007, title={Application of in vivo methodologies to investigation biological structure, function and xenobiotic response in see-through medaka (Oryzias latipes}, volume={11}, journal={The Fish Biology Journal, Medaka}, author={Hardman, R.C. and Kullman, S.W. and Hinton, D.E.}, year={2007}, pages={43–65} }
 @book{kullman_linden_hinton_2007, title={Bioanalytical Analysis Of Natural Estrogens at Concentrated Animal Feed Operations Affecting North Carolina River Waters}, number={376376}, institution={Water Resources Research Institute of North Carolina}, author={Kullman, S.W. and Linden, K.G. and Hinton, D.E.}, year={2007} }
 @article{chen_rosenfeldt_kullman_hinton_linden_2007, title={Biological assessments of a mixture of endocrine disruptors at environmentally relevant concentrations in water following UV/H2O2 oxidation}, volume={376}, ISSN={0048-9697}, url={http://dx.doi.org/10.1016/j.scitotenv.2006.12.051}, DOI={10.1016/j.scitotenv.2006.12.051}, abstractNote={Numerous studies have investigated degradation of individual endocrine disrupting compounds (EDCs) in lab or natural waters. However, natural variations in water matrices and mixtures of EDCs in the environment may confound analysis of the treatment efficiency. Because chemical based analytical methods cannot represent the combined or synergistic activities between water quality parameters and/or the EDC mixtures at environmentally relevant concentrations (μg L− 1-ng L− 1), bioanalytical assessments of residual estrogenic activity in treated water were used to evaluate the performance of the UV based advanced oxidation process for estrogenic contaminants in water. Four EDCs including estradiol (E2), ethinyl estradiol (EE2), bisphenol-A (BPA) and nonylphenol (NP) were spiked individually or as a mixture at μg L− 1-ng L− 1 in laboratory or natural river water. The removal rates of estrogenic activity were quantitatively evaluated by in vitro yeast estrogen screen (YES) and in vivo Vitellogenin (VTG) assays with Japanese medaka fish (Oryzias latipes). UV in combination with 10 ppm H2O2 as an oxidation process was capable of decreasing in vitro and in vivo estrogenic activity, however, in vivo estrogenic activity of the EDC mixture in natural water was not completely removed at UV fluence up to 2000 mJ cm− 2. The removal rates of in vitro estrogenic activity of the EDC mixtures were lower than those observed for single compounds, and slower in natural waters, likely due to lower steady-state concentrations of hydroxyl radicals (•OH) in the presence of •OH scavengers from the water matrix and EDC mixture.}, number={1-3}, journal={Science of The Total Environment}, publisher={Elsevier BV}, author={Chen, Pei-Jen and Rosenfeldt, Erik J. and Kullman, Seth W. and Hinton, David E. and Linden, Karl G.}, year={2007}, month={Apr}, pages={18–26} }
 @article{chen_kullman_hinton_linden_2007, title={Comparisons of polychromatic and monochromatic UV-based treatments of bisphenol-A in water via toxicity assessments}, volume={68}, ISSN={0045-6535}, url={http://dx.doi.org/10.1016/j.chemosphere.2007.02.020}, DOI={10.1016/j.chemosphere.2007.02.020}, abstractNote={Polychromatic ultraviolet irradiation, such as from medium pressure (MP) Hg lamps may enhance the UV degradation of environmental pollutants as compared to low pressure (LP) Hg UV sources emitting monochromatic irradiation. Typically, studies involving destruction of environmental pollutants such as endocrine disrupting compounds (EDCs) are based on measurement of the parent compound decay using analytical chemistry, but such information is insufficient to determine an effective treatment endpoint because the identity and biological activity of many transformation products remain unknown. Bioanalytical methods to assess residual biological activity of a treated water offers one means to compare removal efficiency of EDC activity between MP- and LP-UV lamps under photolysis and UV/H2O2 oxidation. In this study, changes in estrogenic activity of bisphenol-A (BPA) as a function of UV treatment were evaluated using both an in vitro yeast estrogen screen and in vivo vitellogenin assay with Japanese medaka (Oryzias latipes) fish. Decay of BPA parent compound and formation of degradation products were followed using HPLC analysis. Results demonstrated that MP-UV direct photolysis more effectively removed BPA and associated estrogenic activity compared to LP-UV lamps. UV in combination with H2O2 significantly removed estrogenic activity in vitro and in vivo compared to direct photolysis; however, no significant difference in removal rates was found between the two lamps under UV/H2O2 oxidation. Furthermore, the UV/H2O2 process was effective for reducing embryo toxicity of BPA, but resulted in the production of acidic intermediates, causing acute toxicity and delayed hatching in some medaka embryos.}, number={6}, journal={Chemosphere}, publisher={Elsevier BV}, author={Chen, Pei-Jen and Kullman, Seth W. and Hinton, David E. and Linden, Karl G.}, year={2007}, month={Jun}, pages={1041–1049} }
 @article{rosenfeldt_chen_kullman_linden_2007, title={Destruction of estrogenic activity in water using UV advanced oxidation}, volume={377}, ISSN={0048-9697}, url={http://dx.doi.org/10.1016/j.scitotenv.2007.01.096}, DOI={10.1016/j.scitotenv.2007.01.096}, abstractNote={The transformation of the steroidal Endocrine Disrupting Compounds (EDCs), 17-beta-estradiol (E2) and 17-alpha-ethinyl estradiol (EE2) by direct UV photolysis and UV/H(2)O(2) advanced oxidation was studied from the perspective of the removal of estrogenic activity associated with the compounds. First, experiments were performed to link the oxidation of E2 and EE2 with subsequent reduction in estrogenic activity. No statistically significant difference between removal rates was observed, implying that the oxidation products of E2 and EE2 are not as estrogenic (measured by the Yeast Estrogen Screen (YES)) as the parent compounds. Utilizing the YES, 90% removal of estrogenic activity of E2 and EE2 at environmentally relevant concentrations ( approximately 3 microg L(-1)) was achieved using a combination of 5 mg L(-1) H(2)O(2) and a UV fluence of less than 350 mJ cm(-2). Thus, these compounds, when considered at environmentally relevant levels, are significantly degraded at much lower UV fluences than previously thought. A steady state OH radical model was used to predict oxidation of EE2 in laboratory and natural waters.}, number={1}, journal={Science of The Total Environment}, publisher={Elsevier BV}, author={Rosenfeldt, E and Chen, P and Kullman, S and Linden, K}, year={2007}, month={May}, pages={105–113} }
 @article{kashiwada_kameshiro_tatsuta_sugaya_kullman_hinton_goka_2007, title={Estrogenic modulation of CYP3A38, CYP3A40, and CYP19 in mature male medaka (Oryzias latipes)}, volume={145}, ISSN={1532-0456}, url={http://dx.doi.org/10.1016/j.cbpc.2007.01.009}, DOI={10.1016/j.cbpc.2007.01.009}, abstractNote={We examined cytochrome P450 production and activity and circulating hormone concentrations in male medaka exposed to 17β-estradiol (E2) or 17α-ethinylestradiol (EE2). Intraperitoneal injection of E2 at 1, 10, or 100 μg/g-fish completely suppressed CYP3A38 protein production and suppressed CYP3A40 protein levels by 89%, 52%, or 47%, respectively. CYP3A38 and CYP3A40 mRNA expression was unaltered, and CYP3A enzymatic activity initially increased and then decreased with increasing E2 dose. Males co-cultured with females were exposed to a markedly high concentration (43 ng/L) of E2 secreted by females. CYP3A protein levels in co-cultured males were suppressed. Serum testosterone (TE) and 11keto-testosterone levels in co-cultured males were downregulated to 40% of pre-exposure levels. Serum E2 levels increased in co-cultured males or males exposed to EE2. Testicular CYP19, which converts TE to E2, increased by 9.5 times in males exposed to 50 ng/L EE2 and by 21.5 times in those exposed to 100 ng/L EE2. Male medaka exposed to EE2 showed increased serum Vtg levels. Estrogenic exposure induced Vtg production, suppressed CYP3A protein production, downregulated TE metabolism, and enhanced CYP19 activity. Serum E2 endogenously induced by CYP19 could contribute to Vtg induction in male medaka.}, number={3}, journal={Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology}, publisher={Elsevier BV}, author={Kashiwada, Shosaku and Kameshiro, Maiko and Tatsuta, Haruki and Sugaya, Yoshio and Kullman, Seth W. and Hinton, David E. and Goka, Koichi}, year={2007}, month={Apr}, pages={370–378} }
 @book{linden_kullman_2007, title={Impact of UV and UV Endocrine Disrupting Compounds in Water}, number={91175}, institution={American Water Works Association Research Foundation}, author={Linden, K.G. and Kullman, S.W.}, year={2007} }
 @book{kullman_linden_hinton_2007, title={Towards an Innovative DNA Array Technology for Detection of Pharmaceuticals in Reclaimed Water}, institution={Water Environment Research Foundation (WERF) and Environmental Protection agency (EPA)}, author={Kullman, S.W. and Linden, K.G. and Hinton, D.E.}, year={2007} }
 @article{linden_rosenfeldt_kullman_2007, title={UV/H2O2 degradation of endocrine-disrupting chemicals in water evaluated via toxicity assays}, volume={55}, ISSN={0273-1223 1996-9732}, url={http://dx.doi.org/10.2166/wst.2007.423}, DOI={10.2166/wst.2007.423}, abstractNote={Due to rising concern regarding the presence of endocrine-disrupting chemicals (EDCs) in surface water and groundwater throughout the United States, Asia and Europe, treatment of these chemicals in drinking water and wastewater to protect human health and the environment is an area of great interest. Many conventional treatment schemes are relatively ineffective in removing EDCs from water and wastewater. This is concerning because these chemicals are biologically active at very low concentrations and effects of mixtures are relatively unknown.}, number={12}, journal={Water Science and Technology}, publisher={IWA Publishing}, author={Linden, Karl G. and Rosenfeldt, Erik J. and Kullman, Seth W.}, year={2007}, month={Jun}, pages={313–319} }
 @article{chen_linden_hinton_kashiwada_rosenfeldt_kullman_2006, title={Biological assessment of bisphenol A degradation in water following direct photolysis and UV advanced oxidation}, volume={65}, ISSN={0045-6535}, url={http://dx.doi.org/10.1016/j.chemosphere.2006.04.048}, DOI={10.1016/j.chemosphere.2006.04.048}, abstractNote={Endocrine disrupting compounds (EDCs) are exogenous environmental chemicals that can interfere with normal hormone function and present a potential threat to both environmental and human health. The fate, distribution and degradation of EDCs is a subject of considerable investigation. To date, several studies have demonstrated that conventional water treatment processes are ineffective for removal of most EDCs and in some instances produce multiple unknown transformation products. In this study we have investigated the use of direct photolysis with low-pressure (LP) Hg UV lamps and UV + hydrogen peroxide (H2O2) advanced oxidation process (AOP) for the degradation of a prototypic endocrine disrupter, bisphenol A (BPA), in laboratory water. Removal rates of BPA and formation of degradation products were determined by high performance liquid chromatography (HPLC) analysis. Changes in estrogenic activity were evaluated using both in vitro yeast estrogen screen (YES) and in vivo vitellogenin (VTG) assays with Japanese medaka fish (Oryzias latipes). Our results demonstrate that UV alone did not effectively degrade BPA. However, UV in combination with H2O2 significantly removed BPA parent compound and aqueous estrogenic activity in vitro and in vivo. Removal rates of in vivo estrogenic activity were significantly lower than those observed in vitro, demonstrating differential sensitivities of these bioassays and that certain UV/AOP metabolites may retain estrogenic activity. Furthermore, the UV/H2O2 AOP was effective for reducing larval lethality in treated BPA solutions, suggesting BPA degradation occurred and that the degradation process did not result in the production of acutely toxic intermediates.}, number={7}, journal={Chemosphere}, publisher={Elsevier BV}, author={Chen, Pei-Jen and Linden, Karl G. and Hinton, David E. and Kashiwada, Shosaku and Rosenfeldt, Erik J. and Kullman, Seth W.}, year={2006}, month={Nov}, pages={1094–1102} }
 @article{volz_hinton_law_kullman_2006, title={Dynamic Gene Expression Changes Precede Dioxin-Induced Liver Pathogenesis in Medaka Fish}, volume={89}, ISSN={1096-6080 1096-0929}, url={http://dx.doi.org/10.1093/toxsci/kfj033}, DOI={10.1093/toxsci/kfj033}, abstractNote={A major challenge for environmental genomics is linking gene expression to cellular toxicity and morphological alteration. Herein, we address complexities related to hepatic gene expression responses after a single injection of the aryl hydrocarbon receptor (AHR) agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin) and illustrate an initial stress response followed by cytologic and adaptive changes in the teleost fish medaka. Using a custom 175-gene array, we find that overall hepatic gene expression and histological changes are strongly dependent on dose and time. The most pronounced dioxin-induced gene expression changes occurred early and preceded morphologic alteration in the liver. Following a systematic search for putative Ah response elements (AHREs) (5'-CACGCA-3') within 2000 bp upstream of the predicted transcriptional start site, the majority (87%) of genes screened in this study did not contain an AHRE, suggesting that gene expression was not solely dependent on AHRE-mediated transcription. Moreover, in the highest dosage, we observed gene expression changes associated with adaptation that persisted for almost two weeks, including induction of a gene putatively identified as ependymin that may function in hepatic injury repair. These data suggest that the cellular response to dioxin involves both AHRE- and non-AHRE-mediated transcription, and that coupling gene expression profiling with analysis of morphologic pathogenesis is essential for establishing temporal relationships between transcriptional changes, toxicity, and adaptation to hepatic injury.}, number={2}, journal={Toxicological Sciences}, publisher={Oxford University Press (OUP)}, author={Volz, David C. and Hinton, David E. and Law, J. McHugh and Kullman, Seth W.}, year={2006}, month={Feb}, pages={524–534} }
 @article{volz_bencic_hinton_law_kullman_2005, title={2,3,7,8-Tetrachlorodibenzo-p-Dioxin (TCDD) Induces Organ- Specific Differential Gene Expression in Male Japanese Medaka (Oryzias latipes)}, volume={85}, ISSN={1096-6080 1096-0929}, url={http://dx.doi.org/10.1093/toxsci/kfi109}, DOI={10.1093/toxsci/kfi109}, abstractNote={2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous environmental contaminant with well-known adverse effects in fish. In this study, we initially exploited suppression subtractive hybridization (SSH) as a screening tool to assess qualitative gene expression changes in whole brain, liver, and testis of adult male Japanese medaka (Oryzias latipes) exposed for 48 h to a single intraperitoneal-injected dose of TCDD (10 microg TCDD/kg body weight). Across these three organs, SSH identified a total of 335 unique genes. Each set of forward- and reverse-subtracted organ cDNA libraries consisted of a distinct gene list and corresponding distribution of biological processes, suggesting that transcript profiles of these libraries were highly organ-specific. Based on sequence match significance and frequencies within each set of organ libraries, genes hypothesized to be strongly responsive (42 total) within male medaka brain, liver, or testis were semi-quantitatively screened with replicate cDNA nylon membrane arrays. In addition, TCDD-treated male medaka were surveyed for gross histological analysis of brain, liver, and testis. In general, adverse histopathological changes were not observed in the brain, and glycogen depletion was observed only in the liver. However, significant histological changes occurred in the testis, and included disorganization of spermatogenesis at the testis periphery, disruption of the interstitium, Leydig cell swelling, and Sertoli cell vacuolation. Of the 42 genes screened by cDNA array analysis, cytochrome P450 1A (CYP1A) mRNA was the only transcript significantly higher in TCDD-exposed brain, whereas 12 transcripts (including CYP1A) were significantly higher in TCDD-exposed liver, and 34 transcripts were significantly lower in TCDD-exposed testis. Therefore, the degree of TCDD-induced alterations observed in each organ at a gross histological level corresponded well with the number and ontology of gene transcripts affected on the array. Based on real-time reverse transcription polymerase chain reaction (RT-PCR), relative CYP1A (but not AHR1) transcript levels were confirmed to be significantly higher in TCDD-treated brain and liver. However, CYP1A was not significantly induced in TCDD-exposed testis, suggesting that gene expression and histopathological responses observed in the testis at 48 h may be CYP1A-independent. Based on these data, unique liver-specific and testis-specific mRNA-level targets in male medaka were identified as promising biomarkers of acute TCDD-induced toxicity.}, number={1}, journal={Toxicological Sciences}, publisher={Oxford University Press (OUP)}, author={Volz, David C. and Bencic, David C. and Hinton, David E. and Law, J. McHugh and Kullman, Seth W.}, year={2005}, month={Feb}, pages={572–584} }
 @article{kashiwada_hinton_kullman_2005, title={Functional characterization of medaka CYP3A38 and CYP3A40: Kinetics and catalysis by expression in a recombinant baculovirus system}, volume={141}, ISSN={1532-0456}, url={http://dx.doi.org/10.1016/j.cca.2005.07.006}, DOI={10.1016/j.cca.2005.07.006}, abstractNote={Phylogenic analysis of the teleost genomic lineages has demonstrated the precedent for multiple genome duplications. Among many of the genes duplicated, cytochrome P450 genes have undergone independent diversification, which can be traced to a single ancestral gene. In teleosts, cytochrome P450s, from all major families, have been identified. Among these, the CYP3A family has been cloned in several teleost species and demonstrated to contain multiple paralogs differing in gene expression patterns and tissue distribution. Herein we characterized the catalytic and kinetic activities of two medaka CYP3A paralogs (CYP3A38 and CYP3A40) with benzyloxyresorufin (BFC), a fluorescent 3A-selective substrate, and testosterone, a known metabolic substrate for CYP3A enzymes. Recombinant CYP3A was produced using the baculovirus expression vector system in Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tn5) insect cells and accounted for up to 24% of total cellular protein. Following addition of a heme-albumin conjugate to log phase cells, spectral P450 content reached a maximum of 560 and 2350 pmol/mg microsomal protein for CYP3A38 and CYP3A40, respectively. Incubations containing recombinant CYP3A, human NADPH-cytochrome P-450 oxidoreductase reductase, human cytochrome b5, and a NADPH generation system catalyzed the dealkylation of BFC and hydroxylation of testosterone with a high degree of stereoselectivity. However, efficiencies and specificities were significantly different between the two isoforms. Km and Vmax activities based on BFC-catalysis were 0.116 and 0.363 muM, and 7.95 and 7.77 nmol/min/nmol P450 for CYP3A38 and CYP3A40, respectively. CYP3A38 preferentially catalyzed testosterone hydroxylation at the 6beta-, 2beta- and 16beta-positions with minor hydroxylation at other positions within the steroid nucleus. Testosterone catalysis with CYP3A40 was limited predominantly to the 6beta- and 2beta-positions. Putative identification of CYP3A substrate recognition sites (SRS) 1-6 indicates that 12 of the 49 amino acid differences between CYP3A38 and CYP3A40 OFRs occur in SRS regions previously known to be associated with steroid hydroxylation. We suggest that differences in kinetics and catalytic activities are a result of amino acid substitutions in SRS regions 1, 3 and 5 within the CYP3A38 and CYP3A40 protein sequence.}, number={4}, journal={Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology}, publisher={Elsevier BV}, author={Kashiwada, Shosaku and Hinton, David E. and Kullman, Seth W.}, year={2005}, month={Aug}, pages={338–348} }
 @article{hinton_kullman_hardman_volz_chen_carney_bencic_2005, title={Resolving mechanisms of toxicity while pursuing ecotoxicological relevance?}, volume={51}, ISSN={0025-326X}, url={http://dx.doi.org/10.1016/j.marpolbul.2005.07.020}, DOI={10.1016/j.marpolbul.2005.07.020}, abstractNote={In this age of modern biology, aquatic toxicological research has pursued mechanisms of action of toxicants. This has provided potential tools for ecotoxicologic investigations. However, problems of biocomplexity and issues at higher levels of biological organization remain a challenge. In the 1980s and 1990s and continuing to a lesser extent today, organisms residing in highly contaminated field sites or exposed in the laboratory to calibrated concentrations of individual compounds were carefully analyzed for their responses to priority pollutants. Correlation of biochemical and structural analyses in cultured cells and tissues, as well as the in vivo exposures led to the production and application of biomarkers of exposure and effect and to our awareness of genotoxicity and its chronic manifestations, such as neoplasms, in wild fishes. To gain acceptance of these findings in the greater environmental toxicology community, “validation of the model” versus other, better-established often rodent models, was necessary and became a major focus. Resultant biomarkers were applied to heavily contaminated and reference field sites as part of effects assessment and with investigations following large-scale disasters such as oil spills or industrial accidents. Over the past 15 years, in the laboratory, small aquarium fish models such as medaka (Oryzias latipes), zebrafish (Danio rerio), platyfish (Xiphophorus species), fathead minnow (Pimephales promelas), and sheepshead minnow (Cyprinodon variegatus) were increasingly used establishing mechanisms of toxicants. Today, the same organisms provide reliable information at higher levels of biological organization relevant to ecotoxicology. We review studies resolving mechanisms of toxicity and discuss ways to address biocomplexity, mixtures of contaminants, and the need to relate individual level responses to populations and communities.}, number={8-12}, journal={Marine Pollution Bulletin}, publisher={Elsevier BV}, author={Hinton, David E. and Kullman, Seth W. and Hardman, Ron C. and Volz, David C. and Chen, Pei-Jen and Carney, Michael and Bencic, David C.}, year={2005}, month={Jan}, pages={635–648} }
 @article{kullman_kashiwada_hinton_2004, title={Analysis of medaka cytochrome P450 3A homotropic and heterotropic cooperativity}, volume={58}, ISSN={0141-1136}, url={http://dx.doi.org/10.1016/j.marenvres.2004.03.030}, DOI={10.1016/j.marenvres.2004.03.030}, abstractNote={We have previously demonstrated that medaka CYP3A is associated with metabolism of several endobiotics including steroids and bile acids. In this study, we demonstrate that medaka CYP3A catalysis exhibits unusual kinetic behaviors consistent with allosteric interaction which cannot be described by hyperbolic kinetic models. Using 7-benzyloxy-4-(trifluoromethyl)-coumarin (BFC) and nonylphenol as CYP3A substrates, we describe both homotropic and heterotropic cooperative interactions. Given the role of CYP3A in maintaining the homeostatic balance for numerous endobiotics, enzymatic activation/inhibition by endocrine disruptors (EDCs) represents a putative (non-genomic) mechanism for endocrine disruption.}, number={2-5}, journal={Marine Environmental Research}, publisher={Elsevier BV}, author={Kullman, Seth W. and Kashiwada, Shosaku and Hinton, David E.}, year={2004}, month={Aug}, pages={469–473} }
 @misc{johnson_kullman_lin_2004, title={Bioinformatics Tools}, ISBN={9780824754303 9780203025925}, ISSN={2154-7408}, url={http://dx.doi.org/10.1201/9780203025925.ch19}, DOI={10.1201/9780203025925.ch19}, journal={Nutrition and Disease Prevention}, publisher={CRC Press}, author={Johnson, Kimberly and Kullman, Seth and Lin, Simon}, year={2004}, month={Aug}, pages={449–472} }
 @article{kullman_hinton_2003, title={Analysis of medaka CYP3A gene regulation, promoter regulatory regions and cloning of the orphan nuclear receptor PXR}, volume={42}, journal={The MDIBL Bulletin}, author={Kullman, S.W. and Hinton, D.E.}, year={2003}, pages={29–31} }
 @article{kullman_matsumura_hinton_2003, title={Estrogen signaling in trout liver: Activation of the ras p21/MAP-kinase pathway}, volume={10}, journal={Environmental Science}, author={Kullman, S.W. and Matsumura, F. and Hinton, D.E.}, year={2003}, pages={223–227} }
 @article{liu_kullman_bencic_torten_hinton_2003, title={ras oncogene mutations in diethylnitrosamine-induced hepatic tumors in medaka (Oryzias latipes), a teleost fish}, volume={539}, ISSN={1383-5718}, url={http://dx.doi.org/10.1016/s1383-5718(03)00133-5}, DOI={10.1016/s1383-5718(03)00133-5}, abstractNote={Medaka fish are an established non-mammalian research model for the study of liver carcinogenesis and exposure to environmental pollutants. Studies have emphasized the development of hepatic neoplasms in medaka following exposure to model carcinogens. To date however, little information is known regarding the mechanisms underlying initiation of hepatic tumors in this species. The aim of this study was to relate our understanding of diethylnitrosamine (DEN)-induced tumor formation to ras gene activation in hepatic neoplasms of exposed medaka. Initial studies were conducted to identify medaka ras exons 1 and 2 by reverse transcriptase polymerase chain reaction (RT-PCR). Amplification of ras exons 1 and 2 from untreated medaka liver resulted in the identification of three polymorphic ras sequence variants exhibiting a high degree of homology to other teleost and mammalian ras genes. Exposure of medaka to 159 ppm of DEN resulted in a wide range of hepatic neoplasms including: hepatocellular adenomas, hepatocellular carcinomas, cholangiomas, and mixed hepatocholangiocellular carcinomas. Individual liver tumors were examined for oncogenically activating ras mutations by probing genomic DNA with probes specific for activating point mutations or by direct cloning and sequencing of ras transcripts using RT-PCR. Using allele-specific oligonucleotide (ASO) analysis, a single point mutation was detected in codon 12 position two in 8/25 (32%) tumors examined. Mutated ras alleles were additionally detected in 12 of 39 (30%) medaka liver tumors by sequence analysis. Ten of the 12 mutations identified contained a single point mutation at codon 12 resulting in a Gly to Asp amino acid substitution. Two unique mutations were identified at codon 16 resulting in either Lys to Asn or Lys to Thr amino acid substitutions. Our results show that ras mutations are induced by DEN and are present in over 30% of the fish that developed tumors. A ras mutation incidence of 30% is similar to that reported in mammalian species exposed to DEN. While mutations at codon 12 have previously been reported, the present study is the first in vivo report of ras point mutations at codon 16.}, number={1-2}, journal={Mutation Research/Genetic Toxicology and Environmental Mutagenesis}, publisher={Elsevier BV}, author={Liu, Zi and Kullman, Seth W. and Bencic, David C. and Torten, Michael and Hinton, David E.}, year={2003}, month={Aug}, pages={43–53} }
 @article{arukwe_kullman_berg_goksøyr_hinton_2002, title={Molecular cloning of rainbow trout (Oncorhynchus mykiss) eggshell zona radiata protein complementary DNA: mRNA expression in 17β-estradiol- and nonylphenol-treated fish}, volume={132}, ISSN={1096-4959}, url={http://dx.doi.org/10.1016/s1096-4959(02)00009-x}, DOI={10.1016/s1096-4959(02)00009-x}, abstractNote={A complementary DNA (cDNA) encoding the eggshell zona radiata protein (RbtZR: AF407574) has been cloned from the liver of estradiol-17β (E2)-treated rainbow trout (Oncorhynchus mykiss) by reverse-transcriptase polymerase chain reaction (RT-PCR). A set of degenerate primers homologous to the highly conserved cysteine-rich region of the zona radiata protein gene from salmon, winter flounder, medaka and carp were used for the initial RT-PCR. The resulting PCR product was cloned, sequenced and identified as the Zrp gene fragment based on amino acid sequence similarities. Based on the Zrp sequence from the initial PCR, a pair of gene-sequence primers was designed for 3′- and 5′- random amplification of cDNA ends (RACE). Cloning and sequencing of RACE products showed a 1349-bp Zrp gene encoding a 403-amino acid protein with a theoretical molecular mass of approximately 45 kDa. Alignment of the deduced amino acid sequence reveals that RbtZR is similar to piscine and mammalian zona pellucida proteins. The RbtZR gene, together with the estrogen receptor (ER) and vitellogenin (Vtg) genes, was further characterized and comparatively studied for transcriptional and translational expression in xenoestrogen- (nonylphenol, NP) and E2-treated juvenile rainbow trout in a time-course experiment. Northern and slot blot analysis showed that the RbtZR mRNA was expressed, in parallel with the ER and Vtg mRNA, in both NP- and E2-treated juvenile rainbow trout. Indirect enzyme-linked immunosorbent assay (ELISA) using monoclonal antibody raised against Atlantic salmon Zrp indicated the translational expression of RbtZR protein in blood plasma samples from NP- and E2-treated juvenile trout. The differential time-dependent transcriptional and translational expression and use of Zrp, ER and Vtg as sensitive biomarkers in environmental monitoring of endocrine disrupters in fish is discussed.}, number={2}, journal={Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology}, publisher={Elsevier BV}, author={Arukwe, Augustine and Kullman, Seth W. and Berg, Karin and Goksøyr, Anders and Hinton, David E.}, year={2002}, month={Jun}, pages={315–326} }
 @article{kullman_hinton_2002, title={Toxicant induced differential gene expression and production of an aquatic gene array}, volume={41}, journal={The MDIBL Bulletin}, author={Kullman, S.W. and Hinton, D.E.}, year={2002}, pages={58–61} }
 @article{arukwe_kullman_hinton_2001, title={Differential biomarker gene and protein expressions in nonylphenol and estradiol-17β treated juvenile rainbow trout (Oncorhynchus mykiss)}, volume={129}, ISSN={1532-0456}, url={http://dx.doi.org/10.1016/s1532-0456(01)00170-3}, DOI={10.1016/s1532-0456(01)00170-3}, abstractNote={The time- and dose-dependent transcriptional and translational expression of biomarker genes in nonylphenol (NP) and estradiol-17beta (E(2)) treated juvenile rainbow trout is reported. Fish were exposed to NP (1, 5 and 25 mg/kg) and E(2) (5 mg/kg) and killed at 2, 6, 12, 24, 48 and 72 h after exposure. The estrogen receptor (ER), vitellogenin (Vtg) and eggshell zona radiata protein (Zr-protein) gene expressions were analyzed in total liver RNA using Northern and slot hybridization with specific cDNA probes. Plasma Vtg and Zr-protein levels were evaluated using indirect ELISA. While Zr-protein gene showed an induction only at 24 h post-exposure, the plasma protein levels showed a time-dependent increase in the 25-mg NP treated group. Vtg transcripts showed an apparent time-dependent increase without a concomitant increase in protein levels in the 25-mg NP treated fish. Time-dependent increases in Vtg and Zr-protein gene expressions without the corresponding increases in ER gene transcription was observed in E(2)-treated fish at 2, 6 and 12 h post-exposure. Induction of ER gene transcripts was observed from 24 h and did not change significantly at 48 and 72 h. In the E(2)-treated fish, induction of plasma Vtg levels was observed at 48 and 72 h, while plasma Zr-protein was induced at 24, 48 and 72 h, after exposure. We conclude that the E(2)- and NP-induced Vtg and Zr-protein gene expressions at the early time intervals after exposure are not dependent on increase in the transcriptional activity of the ER gene and that Vtg and Zr-protein gene transcriptions require only basal or minimal ER concentration, in addition to other mechanisms.}, number={1}, journal={Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology}, publisher={Elsevier BV}, author={Arukwe, A and Kullman, S.W. and Hinton, D.E.}, year={2001}, month={May}, pages={1–10} }
 @article{kullman_hinton_2001, title={Identification, characterization, and ontogeny of a second cytochrome P450 3A gene from the fresh water teleost medaka (Oryzias latipes)}, volume={58}, ISSN={1040-452X 1098-2795}, url={http://dx.doi.org/10.1002/1098-2795(200102)58:2<149::aid-mrd3>3.0.co;2-x}, DOI={10.1002/1098-2795(200102)58:2<149::aid-mrd3>3.0.co;2-x}, abstractNote={Multiple copies of cytochrome P450 gene family 3 have been identified from numerous mammalian species. Often these genes exhibit differential catalytic activities and gene regulation. To date however, little information is available regarding multiple forms of this gene family in teleost fishes. In this study, a second isozyme of cytochrome P450 3A has been cloned from the teleost fish Oryzias latipes and designated CYP3A40. Screening of a cDNA library to medaka liver resulted in the identification of a full length cDNA clone containing a 2316 base pair (bp) insert with an open reading frame encoding a single peptide of 502 amino acids. Comparisons of the deduced amino acid sequence to other known cytochrome P450 sequences indicate that this gene product is most similar to the CYP3A gene family and shares a 90% identity to CYP3A38 previously identified from medaka liver. Consistent with Northern blot and Western blot analysis, Southern blots of medaka genomic DNA demonstrated the presence of two CYP3A genes. Gene expression studies demonstrated that CYP3A38 and CYP3A40 are differentially regulated according to embryonic development. Northern blot analysis, using a probe to a conserved region of both CYP3A genes, demonstrated the presence of a single CYP3A transcript for early and late embryonic stages and two CYP3A transcripts in larvae and adult liver. Similarly, Western blots show a single faint immunoreactive cytochrome P450 3A protein in microsomes from early and late embryos and two abundant protein bands in microsomes from larval and adult liver. To further examine the transcriptional differences in CYP3A expression, RT‐PCR analysis was performed on embryonic stages 11–35, 1‐ and 14‐day‐old larvae, and adult liver using primer sets specific for CYP3A38 and CYP3A40. These results demonstrate that CYP3A40 is expressed early in embryonic development and continues throughout adult stages. CYP3A38, however, is tightly regulated during embryonic development and is only expressed post‐hatch Mol. Reprod. Dev. 58:149–158, 2001. © 2001 Wiley‐Liss, Inc.}, number={2}, journal={Molecular Reproduction and Development}, publisher={Wiley}, author={Kullman, Seth W. and Hinton, David E.}, year={2001}, month={Feb}, pages={149–158} }
 @article{kullman_hamm_hinton_2000, title={Identification and Characterization of a cDNA Encoding Cytochrome P450 3A from the Fresh Water Teleost Medaka (Oryzias latipes)}, volume={380}, ISSN={0003-9861}, url={http://dx.doi.org/10.1006/abbi.2000.1904}, DOI={10.1006/abbi.2000.1904}, abstractNote={A new member of the CYP3A gene family has been cloned from the teleost fish medaka (Oryzias latipes) by reverse-transcriptase polymerase chain reaction (RT-PCR). Degenerate primers homologous to highly conserved regions of known CYP3A sequences were used for initial RT-PCRs. Individual PCR products were cloned, sequenced, and identified as those belonging to the cytochrome P450 superfamily based on amino acid sequence similarity and the presence of the highly conserved heme-binding region. PCR products were subsequently used as probes to screen a complementary DNA library. A full-length cDNA clone was identified containing a 1758-base-pair (bp) insert with an open reading frame encoding a single peptide of 500 amino acids. Comparisons of the deduced amino acid sequence to other known cytochrome P450 sequences indicate that this gene product is most similar to the CYP3A gene family and has been designated as CYP3A38 by the cytochrome P450 nomenclature committee. Northern blot analysis identified two abundant CYP3A related transcripts in liver of both male and female adults and demonstrated quantitative differences in abundance according to gender. Similarly, Western blot analysis demonstrated the presence of two abundant cytochrome P450 related proteins in liver of both male and female adults. These results suggests that O. latipes contains multiple forms of CYP3A. Heterologous expression of CYP3A38 cDNA in HEK 293 cells produced a single protein that was reactive with anti-scup P450A (CYP3A) polyclonal antibody. Microsomes of HEK 293 cells expressing recombinant CYP3A38 protein actively catalyzed the hydroxylation of testosterone.}, number={1}, journal={Archives of Biochemistry and Biophysics}, publisher={Elsevier BV}, author={Kullman, Seth W. and Hamm, Jonathan T. and Hinton, David E.}, year={2000}, month={Aug}, pages={29–38} }
 @article{kullman_matsumura_1997, title={Identification of a novel cytochrome P-450 gene from the white rot fungus Phanerochaete chrysosporium}, volume={63}, ISSN={0099-2240 1098-5336}, url={http://dx.doi.org/10.1128/aem.63.7.2741-2746.1997}, DOI={10.1128/aem.63.7.2741-2746.1997}, abstractNote={A gene fragment belonging to the cytochrome P-450 superfamily has been cloned and identified from stationary cultures of the filamentous fungus Phanerochaete chrysosporium by reverse transcriptase (RT)-PCR. A set of degenerate primers homologous to highly conserved regions of known cytochrome P-450 sequences were used for initial RT-PCRs. Individual PCR products were cloned, sequenced, and identified as those belonging to the cytochrome P-450 superfamily based on amino acid sequence homologies and the presence of the highly conserved heme binding region. The nucleotide sequence of a single cDNA clone indicated the presence of an open reading frame encoding a partial cytochrome P-450 protein of 208 amino acids. Comparisons of the deduced amino acid sequence of the partial protein to other known cytochrome P-450 sequences indicate that it is the first member of a new family of cytochrome P-450s, designated CYP63-1A. Northern blot analysis suggests that CYP63-1A is expressed under both nitrogen-rich and nitrogen-deficient culture conditions and thus not under the same regulatory constraints as the well-studied lignin and manganese peroxidases. Western blot analyses using antibodies raised to the heme binding region of CYP63-1A indicate that the protein has a molecular mass of approximately 44,000 Da.}, number={7}, journal={Applied and Environmental Microbiology}, publisher={American Society for Microbiology}, author={Kullman, S W and Matsumura, F}, year={1997}, month={Jul}, pages={2741–2746} }
 @article{kullman_matsumura_1996, title={Metabolic pathways utilized by Phanerochaete chrysosporium for degradation of the cyclodiene pesticide endosulfan}, volume={62}, ISSN={0099-2240 1098-5336}, url={http://dx.doi.org/10.1128/aem.62.2.593-600.1996}, DOI={10.1128/aem.62.2.593-600.1996}, abstractNote={Recent studies have shown that cultures of white rot fungi not favoring the production of lignin and manganese peroxidases are effective in degrading certain xenobiotics. In this study we have used endosulfan as a model xenobiotic to assess the enzymatic mechanisms of pesticide metabolism under ligninolytic (nutrient-deficient) and nonligninolytic (nutrient-rich) culture conditions. Rapid metabolism of this chlorinated pesticide occurred under each nutrient condition tested. However, the extent of degradation and the nature of the metabolic products differed for nutrient-deficient and nutrient-rich media. The pathways for endosulfan metabolism were characterized by analysis of the fungal metabolites produced. The major endosulfan metabolites were identified by gas chromatography-electron capture detection and gas chromatography-mass spectrometry as endosulfan sulfate, endosulfan diol, endosulfan hydroxyether, and a unknown metabolite tentatively identified as endosulfan dialdehyde. The nature of the metabolites formed indicates that this organism utilizes both oxidative and hydrolytic pathways for metabolism of this pesticide. Piperonyl butoxide, a known cytochrome P-450 inhibitor, significantly inhibited the oxidation of endosulfan to endosulfan sulfate and enhanced hydrolysis of endosulfan to endosulfan diol. We suggest that the metabolism of endosulfan is mediated by two divergent pathways, one hydrolytic and the other oxidative. Judging by the inactivity of extracellular fluid and partially purified lignin peroxidase in metabolizing endosulfan, we conclude that metabolism of this compound does not involve the action of extracellular peroxidases.}, number={2}, journal={Applied and Environmental Microbiology}, publisher={American Society for Microbiology}, author={Kullman, S W and Matsumura, F}, year={1996}, month={Feb}, pages={593–600} }
 @book{kullman_lindenauer_fuller_1995, title={Bioremediation Principles: Laboratory Manual}, institution={Department of Civil and Environmental Engineering and US Department of Defense, Bioremediation course development for Marie Island Remediation}, author={Kullman, S.W. and Lindenauer, K.G. and Fuller, M.}, year={1995}, month={Jul} }
 @article{lucas_bekheit_goodrow_jones_kullman_matsumura_woodrow_seiber_hammock_1993, title={Development of antibodies against hydroxyatrazine and hydroxysimazine: Application to environmental samples}, volume={41}, ISSN={0021-8561 1520-5118}, url={http://dx.doi.org/10.1021/jf00033a032}, DOI={10.1021/jf00033a032}, abstractNote={An enzyme-linked immunosorbent assay (ELISA) selective for hydroxyatrazine and hydroxysimazine was developed as a means of monitoring environmental and microbial degradation of atrazine to hydroxyatrazine. This ELISA was tolerant to solvents, salts, and pH changes and functioned well in aseries of matrices (water, soil, horse manure, urine, and fungal extracts). In preliminary bioremediation studies conducted in a drum impervious to UV light, the microbes endogenous to horse manure converted approximately 1% of the total initial amount of atrazine to hydroxyatrazine. Also, a UV-resistant strain of white rot fungus (Phanerochaete chrysosporium) was tested for ita ability to degrade atrazine. Using the ELISA described here, approximately half of the time zero amount of atrazine (2 pg/35-mm Petri dish) was gone in 1 day and essentially all of the atrazine was converted to hydroxyatrazine, primarily due to UV irradiation. This ELISA provided a valuable method to quantify hydroxyatrazine and hydroxysimazine in a series of traditionally difficult matrices for the investigation of the bioremediation of atrazine.}, number={9}, journal={Journal of Agricultural and Food Chemistry}, publisher={American Chemical Society (ACS)}, author={Lucas, Anne D. and Bekheit, Hassan K. M. and Goodrow, Marvin H. and Jones, A. Daniel. and Kullman, Seth. and Matsumura, Fumio. and Woodrow, James E. and Seiber, James N. and Hammock, Bruce D.}, year={1993}, month={Sep}, pages={1523–1529} }