Application of Evolving Computational and Biological Platforms for Chemical Safety Assessment

Abstract The 21st century is placing new demands on toxicology, such as the need to evaluate many more chemicals than ever before, and to make toxicity testing more relevant to human exposure. Concomitantly, advances in molecular biology, systems biology, and computational methods are available to address these new needs. As a result, toxicology finds itself in a major transformational period represented by a shift from an observational science to one that is more predictive. The intent of this chapter is to review the enormously wide range of computational and biological assay platforms that are available for use in constructing a predictive toxicology program. These assay platforms all have their own strengths and weaknesses that need to be carefully considered. We conclude with a discussion of potential applications of these assay platforms.

[1]  I Kimber,et al.  The murine local lymph node assay: a commentary on collaborative studies and new directions. , 1992, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[2]  George Daston,et al.  Framework for identifying chemicals with structural features associated with the potential to act as developmental or reproductive toxicants. , 2013, Chemical research in toxicology.

[3]  Tao Chen,et al.  Mutant frequency and mutational spectra in the Tk and Hprt genes of N‐ethyl‐N‐nitrosourea‐treated mouse lymphoma cells † , 2002, Environmental and molecular mutagenesis.

[4]  Richard A Becker,et al.  Read-across approaches--misconceptions, promises and challenges ahead. , 2014, ALTEX.

[5]  A. Guillouzo,et al.  Liver cell models in in vitro toxicology. , 1998, Environmental health perspectives.

[6]  Mark Johnston,et al.  Whither Model Organism Research? , 2005, Science.

[7]  Robert J Kavlock,et al.  Incorporating human dosimetry and exposure into high-throughput in vitro toxicity screening. , 2010, Toxicological sciences : an official journal of the Society of Toxicology.

[8]  Catherine Ribault,et al.  Highly efficient gene transfer into hepatocyte‐like HepaRG cells: New means for drug metabolism and toxicity studies , 2010, Biotechnology journal.

[9]  A. Guillouzo,et al.  Comparison of caffeine metabolism by slices, microsomes and hepatocyte cultures from adult human liver. , 1989, Xenobiotica; the fate of foreign compounds in biological systems.

[10]  Richard A Becker,et al.  Toxicity testing in the 21st century: a view from the chemical industry. , 2009, Toxicological sciences : an official journal of the Society of Toxicology.

[11]  A. J. Gandolfi,et al.  Precision-cut tissue chips as an in vitro toxicology system. , 2007, Toxicology in vitro : an international journal published in association with BIBRA.

[12]  A. Kligman,et al.  The Identification of Contact Allergens by Animal Assay. the Guinea Pig Maximization Test , 1969 .

[13]  Hannu Komulainen,et al.  Bacterial and mammalian‐cell genotoxicity of mixtures of chlorohydroxyfuranones, by‐products of water chlorination , 2004, Environmental and molecular mutagenesis.

[14]  D. Massaro,et al.  Mitogenic response of rat lung to endotoxin exposure. , 1987, Biochemical pharmacology.

[15]  A. Y. Lu,et al.  N-acetyl-p-benzoquinone imine: a cytochrome P-450-mediated oxidation product of acetaminophen. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[16]  Yuri Dancik,et al.  Bayesian integrated testing strategy to assess skin sensitization potency: from theory to practice , 2013, Journal of applied toxicology : JAT.

[17]  Erwin van Vliet,et al.  Current standing and future prospects for the technologies proposed to transform toxicity testing in the 21st century. , 2011 .

[18]  J. Chiang,et al.  Transcriptional regulation of the human cholesterol 7 alpha-hydroxylase gene (CYP7A) in HepG2 cells. , 1996, Journal of lipid research.

[19]  D B Dusenbery,et al.  Using the Nematode Caenorhabditis Elegans To Predict Mammalian Acute Lethality To Metallic Salts , 1988, Toxicology and industrial health.

[20]  G Frank Gerberick,et al.  Development of a peptide reactivity assay for screening contact allergens. , 2004, Toxicological sciences : an official journal of the Society of Toxicology.

[21]  P. Wertz,et al.  The human epidermis models EpiSkin, SkinEthic and EpiDerm: an evaluation of morphology and their suitability for testing phototoxicity, irritancy, corrosivity, and substance transport. , 2005, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[22]  D B Dusenbery,et al.  A Promising Indicator of Neurobehavioral Toxicity Using the Nematode Caenorhabditis Elegans and Computer Tracking , 1990, Toxicology and industrial health.

[23]  R. Nagel DarT: The embryo test with the Zebrafish Danio rerio--a general model in ecotoxicology and toxicology. , 2002, ALTEX.

[24]  Stuart L Schreiber,et al.  Chemical suppression of a genetic mutation in a zebrafish model of aortic coarctation , 2004, Nature Biotechnology.

[25]  Thomas Braunbeck,et al.  Towards an alternative for the acute fish LC(50) test in chemical assessment: the fish embryo toxicity test goes multi-species -- an update. , 2005, ALTEX.

[26]  Julieta M. Panzica-Kelly,et al.  Development of a zebrafish embryo teratogenicity assay and quantitative prediction model. , 2010, Birth defects research. Part B, Developmental and reproductive toxicology.

[27]  H Spielmann,et al.  Interlaboratory assessment of the bovine corneal opacity and permeability (BCOP) assay. , 1994, Toxicology in vitro : an international journal published in association with BIBRA.

[28]  R. Judson,et al.  High throughput heuristics for prioritizing human exposure to environmental chemicals. , 2014, Environmental science & technology.

[29]  Hilda Witters,et al.  The ReProTect Feasibility Study, a novel comprehensive in vitro approach to detect reproductive toxicants. , 2010, Reproductive toxicology.

[30]  Aldert Piersma,et al.  The Practical Application of Three Validated In Vitro Embryotoxicity Tests , 2006, Alternatives to laboratory animals : ATLA.

[31]  Thomas Hartung,et al.  Re-evaluation of animal numbers and costs for in vivo tests to accomplish REACH legislation requirements for chemicals - a report by the transatlantic think tank for toxicology (t(4)). , 2009, ALTEX.

[32]  Ivan Rusyn,et al.  Toxicogenetics: population-based testing of drug and chemical safety in mouse models. , 2010, Pharmacogenomics.

[33]  Nicholas Ball,et al.  The challenge of using read-across within the EU REACH regulatory framework; how much uncertainty is too much? Dipropylene glycol methyl ether acetate, an exemplary case study. , 2014, Regulatory toxicology and pharmacology : RTP.

[34]  R. Young,et al.  Stem Cells, the Molecular Circuitry of Pluripotency and Nuclear Reprogramming , 2008, Cell.

[35]  C. Fraga,et al.  Halogenated compounds as inducers of lipid peroxidation in tissue slices. , 1987, Free radical biology & medicine.

[36]  Urs A. Boelsterli,et al.  Biomechanisms of cocaine-induced hepatocyte injury mediated by the formation of reactive metabolites , 2005, Archives of Toxicology.

[37]  I Gerner,et al.  Interlaboratory assessment of alternatives to the Draize eye irritation test in Germany. , 1991, Toxicology in vitro : an international journal published in association with BIBRA.

[38]  Keith B Tierney,et al.  Behavioural assessments of neurotoxic effects and neurodegeneration in zebrafish. , 2011, Biochimica et biophysica acta.

[39]  Michael Boutros,et al.  The art and design of genetic screens: RNA interference , 2008, Nature Reviews Genetics.

[40]  David M. Reif,et al.  Zebrafish developmental screening of the ToxCast™ Phase I chemical library. , 2012, Reproductive toxicology.

[41]  J A Miller,et al.  DNA repair in primary cultures of rat hepatocytes. , 1978, Cancer research.

[42]  M Mizell,et al.  The aquatic vertebrate embryo as a sentinel for toxins: zebrafish embryo dechorionation and perivitelline space microinjection. , 1997, The International journal of developmental biology.

[43]  Phillip L. Williams,et al.  Predicting relative metal toxicity with ion characteristics: Caenorhabditis elegans LC50 , 1997 .

[44]  Jon D Larson,et al.  Morpholino phosphorodiamidate oligonucleotides in zebrafish: a recipe for functional genomics? , 2002, Briefings in functional genomics & proteomics.

[45]  Christina Magkoufopoulou,et al.  Comparison of HepG2 and HepaRG by whole-genome gene expression analysis for the purpose of chemical hazard identification. , 2010, Toxicological sciences : an official journal of the Society of Toxicology.

[46]  Phillip L. Williams,et al.  The nematode Caenorhabditis elegans as a model of organophosphate-induced mammalian neurotoxicity. , 2004, Toxicology and applied pharmacology.

[47]  Aldert H Piersma,et al.  Chemical class-specific gene expression changes in the zebrafish embryo after exposure to glycol ether alkoxy acids and 1,2,4-triazole antifungals. , 2011, Reproductive toxicology.

[48]  Marc Peschanski,et al.  Human pluripotent stem cells in drug discovery and predictive toxicology. , 2010, Biochemical Society transactions.

[49]  G. Serbedzija,et al.  Zebrafish angiogenesis: A new model for drug screening , 2004, Angiogenesis.

[50]  J. Hook,et al.  Mechanism of chloroform nephrotoxicity. II. In vitro evidence for renal metabolism of chloroform in mice. , 1983, Toxicology and applied pharmacology.

[51]  B. Fleischmann,et al.  The use of quantitative image analysis in the assessment of in vitro embryotoxicity endpoints based on a novel embryonic stem cell clone with endoderm-related GFP expression. , 2002, Toxicology in vitro : an international journal published in association with BIBRA.

[52]  Gerberick Frank,et al.  The intra- and inter-laboratory reproducibility and predictivity of the KeratinoSens assay to predict skin sensitizers in vitro: results of a ring-study in five laboratories. , 2011, Toxicology in vitro : an international journal published in association with BIBRA.

[53]  G Patlewicz,et al.  Building scientific confidence in the development and evaluation of read-across. , 2015, Regulatory toxicology and pharmacology : RTP.

[54]  O Pelkonen,et al.  Predictive value of animal models for human cytochrome P450 (CYP)-mediated metabolism: A comparative study in vitro , 2007, Xenobiotica; the fate of foreign compounds in biological systems.

[55]  R. Pulak,et al.  Techniques for analysis, sorting, and dispensing of C. elegans on the COPAS flow-sorting system. , 2006, Methods in molecular biology.

[56]  Hidekazu Tomimoto,et al.  Thalidomide-induced antiangiogenic action is mediated by ceramide through depletion of VEGF receptors, and is antagonized by sphingosine-1-phosphate. , 2005, Blood.

[57]  Daniel Vallero,et al.  SHEDS-HT: an integrated probabilistic exposure model for prioritizing exposures to chemicals with near-field and dietary sources. , 2014, Environmental science & technology.

[58]  Aldert H Piersma,et al.  Validation of alternative methods for developmental toxicity testing. , 2003, Toxicology letters.

[59]  Aldert Piersma,et al.  A Review of the Implementation of the Embryonic Stem Cell Test (EST) , 2009, Alternatives to laboratory animals : ATLA.

[60]  R Zamora,et al.  Damage to protein synthesis concurrent with lipid peroxidation in rat liver slices: effect of halogenated compounds, peroxides, and vitamin E1. , 1989, Archives of biochemistry and biophysics.

[61]  P. Friedman,et al.  Competition between paraquat and putrescine for accumulation by rat lung slices. , 1983, Toxicology.

[62]  E. Alexandre,et al.  Isolation and culture of primary hepatocytes from resected human liver tissue. , 2010, Methods in molecular biology.

[63]  Jeroen L A Pennings,et al.  Identification by gene coregulation mapping of novel genes involved in embryonic stem cell differentiation. , 2011, Stem cells and development.

[64]  Fuart Gatnik Mojca,et al.  Review of QSAR Models and Software Tools for Predicting of Genotoxicity and Carcinogenicity , 2010 .

[65]  Charles Keller,et al.  Refinement of a morphological scoring system for postimplantation rabbit conceptuses. , 2007, Birth defects research. Part B, Developmental and reproductive toxicology.

[66]  Melvin E. Andersen,et al.  Incorporating New Technologies Into Toxicity Testing and Risk Assessment: Moving From 21st Century Vision to a Data-Driven Framework , 2013, Toxicological sciences : an official journal of the Society of Toxicology.

[67]  Richard A Becker,et al.  Tiered toxicity testing: evaluation of toxicity-based decision triggers for human health hazard characterization. , 2007, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[68]  Calum A. MacRae,et al.  Drugs That Induce Repolarization Abnormalities Cause Bradycardia in Zebrafish , 2003, Circulation.

[69]  Stefan O Mueller,et al.  Effects of cell culture conditions on primary rat hepatocytes-cell morphology and differential gene expression. , 2006, Toxicology.

[70]  A. Fleming,et al.  Accumulation and metabolism of drugs and CYP probe substrates in zebrafish larvae , 2010, Xenobiotica; the fate of foreign compounds in biological systems.

[71]  S Fedoroff Proposed usage of animal tissue culture terms. , 1967, Journal of the National Cancer Institute.

[72]  M M Moore,et al.  The L5178Y/tk+/- mouse lymphoma specific gene and chromosomal mutation assay a phase III report of the U.S. Environmental Protection Agency Gene-Tox Program. , 1997, Mutation research.

[73]  A. Rubinstein,et al.  Neuroprotection of MPTP-induced toxicity in zebrafish dopaminergic neurons. , 2005, Brain research. Molecular brain research.

[74]  S. Brenner,et al.  The structure of the ventral nerve cord of Caenorhabditis elegans. , 1976, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[75]  Robert Combes,et al.  Integrated Decision-tree Testing Strategies for Developmental and Reproductive Toxicity with Respect to the Requirements of the EU REACH Legislation , 2008, Alternatives to laboratory animals : ATLA.

[76]  I. Lim,et al.  TIS21/BTG2/PC3 as a link between ageing and cancer: cell cycle regulator and endogenous cell death molecule , 2006, Journal of Cancer Research and Clinical Oncology.

[77]  Chia-Cheng Chang,et al.  Factors to consider in the use of stem cells for pharmaceutic drug development and for chemical safety assessment. , 2010, Toxicology.

[78]  H. Spielmann,et al.  The embryonic stem cell test (EST), an in vitro embryotoxicity test using two permanent mouse cell lines : 3T3 fibroblasts and embryonic stem cells , 1997 .

[79]  A. Guillouzo,et al.  Cultured human adult hepatocytes: a new model for drug metabolism studies. , 1983, Biochemical pharmacology.

[80]  Hilda Witters,et al.  Zebrafish embryos as an alternative to animal experiments--a commentary on the definition of the onset of protected life stages in animal welfare regulations. , 2012, Reproductive toxicology.

[81]  P. Maurel,et al.  Cyclosporin A drug interactions. Screening for inducers and inhibitors of cytochrome P-450 (cyclosporin A oxidase) in primary cultures of human hepatocytes and in liver microsomes. , 1990, Drug metabolism and disposition: the biological fate of chemicals.

[82]  Peter Friedl,et al.  Compensation mechanism in tumor cell migration , 2003, The Journal of cell biology.

[83]  Jörg C Gerlach,et al.  HepaRG human hepatic cell line utility as a surrogate for primary human hepatocytes in drug metabolism assessment in vitro. , 2011, Journal of pharmacological and toxicological methods.

[84]  J. S. Sidhu,et al.  Phenobarbital responsiveness as a uniquely sensitive indicator of hepatocyte differentiation status: requirement of dexamethasone and extracellular matrix in establishing the functional integrity of cultured primary rat hepatocytes. , 2004, Experimental cell research.

[85]  A. Guillouzo,et al.  Species differences in metabolism of ketotifen in rat, rabbit and man: demonstration of similar pathways in vivo and in cultured hepatocytes. , 1987, Life sciences.

[86]  Volker Mersch-Sundermann,et al.  Genotoxic effects of dietary and lifestyle related carcinogens in human derived hepatoma (HepG2, Hep3B) cells. , 2004, Mutation research.

[87]  B P Schmid,et al.  Xenobiotic influences on embryonic differentiation, growth and morphology in vitro. , 1985, Xenobiotica; the fate of foreign compounds in biological systems.

[88]  H B Koëter,et al.  Justification of the enucleated eye test with eyes of slaughterhouse animals as an alternative to the Draize eye irritation test with rabbits. , 1993, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[89]  M. Roberts,et al.  A model to study intestinal and hepatic metabolism of propranolol in the dog. , 2004, Journal of veterinary pharmacology and therapeutics.

[90]  Melvin E. Andersen,et al.  Incorporating High-Throughput Exposure Predictions With Dosimetry-Adjusted In Vitro Bioactivity to Inform Chemical Toxicity Testing , 2015, Toxicological sciences : an official journal of the Society of Toxicology.

[91]  G. Patlewicz,et al.  An evaluation of the implementation of the Cramer classification scheme in the Toxtree software , 2008, SAR and QSAR in environmental research.

[92]  T Takahashi,et al.  Superoxide is involved in the pathogenesis of paraquat‐induced injury in cultured rat liver slices , 1991, Hepatology.

[93]  Patricia McGrath,et al.  Zebrafish: a preclinical model for drug screening. , 2002, Assay and drug development technologies.

[94]  Zheng Zhou,et al.  Detecting apoptotic cells and monitoring their clearance in the nematode Caenorhabditis elegans. , 2009, Methods in molecular biology.

[95]  D L Story,et al.  Response of isolated hepatocytes to organic and inorganic cytotoxins. , 1983, Journal of toxicology and environmental health.

[96]  J. S. Wassom,et al.  A review and analysis of the Chinese hamster ovary/hypoxanthine guanine phosphoribosyl transferase assay to determine the mutagenicity of chemical agents. A report of phase III of the U.S. Environmental Protection Agency Gene-Tox Program. , 1988, Mutation research.

[97]  Marina Marinovich,et al.  Isoeugenol destabilizes IL-8 mRNA expression in THP-1 cells through induction of the negative regulator of mRNA stability tristetraprolin , 2011, Archives of Toxicology.

[98]  Kiyoshi Nagata,et al.  Selective suppressions of human CYP3A forms, CYP3A5 and CYP3A7, by troglitazone in HepG2 cells. , 2002, Drug metabolism and pharmacokinetics.

[99]  Lixin Yang,et al.  Transcriptional profiling reveals barcode-like toxicogenomic responses in the zebrafish embryo , 2007, Genome Biology.

[100]  Shingo Niimi,et al.  Spheroid formation and expression of liver-specific functions of human hepatocellular carcinoma-derived FLC-4 cells cultured in lactose-silk fibroin conjugate sponges. , 2011, Biomacromolecules.

[101]  Olivier Fardel,et al.  Functional expression of sinusoidal and canalicular hepatic drug transporters in the differentiated human hepatoma HepaRG cell line. , 2006, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[102]  E. Little,et al.  Cholinergic and behavioral neurotoxicity of carbaryl and cadmium to larval rainbow trout (Oncorhynchus mykiss). , 2001, Ecotoxicology and environmental safety.

[103]  David M. Reif,et al.  High-throughput models for exposure-based chemical prioritization in the ExpoCast project. , 2013, Environmental science & technology.

[104]  Aldert H Piersma,et al.  Relative embryotoxicity of two classes of chemicals in a modified zebrafish embryotoxicity test and comparison with their in vivo potencies. , 2011, Toxicology in vitro : an international journal published in association with BIBRA.

[105]  A. J. Gandolfi,et al.  Culturing precision-cut human prostate slices as an in vitro model of prostate pathobiology , 2004, Cell Biology and Toxicology.

[106]  L F Stankowski,et al.  An evaluation of the roles of mammalian cell mutation assays in the testing of chemical genotoxicity. , 1991, Regulatory toxicology and pharmacology : RTP.

[107]  Tommy B Andersson,et al.  Evaluation of HepaRG Cells as an in Vitro Model for Human Drug Metabolism Studies , 2008, Drug Metabolism and Disposition.

[108]  L. Cicurel,et al.  Postimplantation embryo culture for the assessment of the teratogenic potential and potency of compounds , 1988, Experientia.

[109]  Marina Marinovich,et al.  Use of IL-18 production in a human keratinocyte cell line to discriminate contact sensitizers from irritants and low molecular weight respiratory allergens. , 2009, Toxicology in vitro : an international journal published in association with BIBRA.

[110]  Mike J. Mason,et al.  Induced pluripotent stem cells and embryonic stem cells are distinguished by gene expression signatures. , 2009, Cell stem cell.

[111]  G Williamson,et al.  Use of metabolically competent human hepatoma cells for the detection of mutagens and antimutagens. , 1998, Mutation research.

[112]  Worth Andrew,et al.  Review of QSAR Models and Software Tools for predicting Developmental and Reproductive Toxicity , 2010 .

[113]  E. S. Wright,et al.  Effects of chronic exposure to ozone on collagen in rat lung. , 1988, Toxicology and applied pharmacology.

[114]  J. Picard,et al.  Whole embryo culture of presomitic mouse embryos. , 1995, Toxicology in vitro : an international journal published in association with BIBRA.

[115]  M G Cherian,et al.  Renal glutathione depletion and nephrotoxicity of cadmium-metallothionein in rats. , 1989, Toxicology and applied pharmacology.

[116]  Toshio Miki,et al.  Hepatic differentiation of human embryonic stem cells is promoted by three-dimensional dynamic perfusion culture conditions. , 2011, Tissue engineering. Part C, Methods.

[117]  R Sanyal,et al.  Inhibition of the genotoxic effects of heterocyclic amines in human derived hepatoma cells by dietary bioantimutagens. , 1997, Mutagenesis.

[118]  R. Judson,et al.  The Toxicity Data Landscape for Environmental Chemicals , 2008, Environmental health perspectives.

[119]  Aldert Piersma,et al.  The ECVAM International Validation Study on In Vitro Embryotoxicity Tests: Results of the Definitive Phase and Evaluation of Prediction Models , 2002, Alternatives to laboratory animals : ATLA.

[120]  M K Prinsen,et al.  The chicken enucleated eye test (CEET): a practical (pre)screen for the assessment of eye irritation/corrosion potential of test materials. , 1996, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[121]  Martha M. Moore,et al.  Use of DNA purified in situ from cells embedded in agarose plugs for the molecular analysis of tk-/- mutants recovered in the L5178Y tk+/- 3.7.2C mutagen assay system. , 1990 .

[122]  A. Guillouzo,et al.  Assessment of the genotoxic potential of indirect chemical mutagens in HepaRG cells by the comet and the cytokinesis-block micronucleus assays. , 2010, Mutagenesis.

[123]  Kristina Luthman,et al.  Allergic contact dermatitis--formation, structural requirements, and reactivity of skin sensitizers. , 2008, Chemical research in toxicology.

[124]  G. Williams,et al.  Detection of chemical carcinogens by unscheduled DNA synthesis in rat liver primary cell cultures. , 1977, Cancer research.

[125]  Roland Buesen,et al.  Trends in improving the embryonic stem cell test (EST): an overview. , 2004, ALTEX.

[126]  Michael O. Hengartner,et al.  Finding function in novel targets: C. elegans as a model organism , 2006, Nature Reviews Drug Discovery.

[127]  André Guillouzo,et al.  The human hepatoma HepaRG cells: a highly differentiated model for studies of liver metabolism and toxicity of xenobiotics. , 2007, Chemico-biological interactions.

[128]  Carl Westmoreland,et al.  A future approach to measuring relative skin sensitising potency: a proposal , 2006, Journal of applied toxicology : JAT.

[129]  Andrea E M Seiler,et al.  Use of murine embryonic stem cells in embryotoxicity assays: the embryonic stem cell test. , 2006, Methods in molecular biology.

[130]  D. Clive,et al.  Laboratory procedure for assessing specific locus mutations at the TK locus in cultured L5178Y mouse lymphoma cells. , 1975, Mutation research.

[131]  S Kato,et al.  An in vitro alternative to the Draize eye-irritation test: Evaluation of the crystal violet staining method. , 1991, Toxicology in vitro : an international journal published in association with BIBRA.

[132]  T H Shepard,et al.  Teratogenic bioactivation of cyclophosphamide in vitro. , 1979, Life sciences.

[133]  Robert E Chapin,et al.  Assessment of the Embryonic Stem Cell Test and application and use in the pharmaceutical industry. , 2008, Birth defects research. Part B, Developmental and reproductive toxicology.

[134]  R Clothier,et al.  The Relative Semi-quantification of mRNA Expression as a Useful Toxicological Endpoint for the Identification of Embryotoxic/Teratogenic Substances. , 1999, Toxicology in vitro : an international journal published in association with BIBRA.

[135]  R A Ford,et al.  Correlation of structural class with no-observed-effect levels: a proposal for establishing a threshold of concern. , 1996, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[136]  M Liebsch,et al.  The ECVAM Prevalidation Study on the Use of EpiDerm for Skin Corrosivity Testing , 2000, Alternatives to laboratory animals : ATLA.

[137]  Christian Trépo,et al.  Origin and characterization of a human bipotent liver progenitor cell line. , 2004, Gastroenterology.

[138]  R. Tompkins,et al.  Long‐Term in Vitro Function of Adult Hepatocytes in a Collagen Sandwich Configuration , 1991, Biotechnology progress.

[139]  J. Caldwell,et al.  Perspective on the usefulness of the mouse lymphoma assay as an indicator of a genotoxic carcinogen: ten compounds which are positive in the mouse lymphoma assay but are not genotoxic carcinogens. , 1993, Teratogenesis, carcinogenesis, and mutagenesis.

[140]  Roger van Egmond,et al.  The Use of the Zebrafish (Danio rerio) Embryo for the Acute Toxicity Testing of Surfactants, as a Possible Alternative to the Acute Fish Test , 2010, Alternatives to laboratory animals : ATLA.

[141]  Marjolein V. Smith,et al.  Medium- and high-throughput screening of neurotoxicants using C. elegans. , 2010, Neurotoxicology and teratology.

[142]  André Guillouzo,et al.  EXPRESSION OF CYTOCHROMES P450, CONJUGATING ENZYMES AND NUCLEAR RECEPTORS IN HUMAN HEPATOMA HepaRG CELLS , 2006, Drug Metabolism and Disposition.

[143]  Windy A. Boyd,et al.  Caenorhabditis elegans generates biologically relevant levels of genotoxic metabolites from aflatoxin B1 but not benzo[a]pyrene in vivo. , 2010, Toxicological sciences : an official journal of the Society of Toxicology.

[144]  Diego A. Golombek,et al.  An automated tracking system for Caenorhabditis elegans locomotor behavior and circadian studies application , 2007, Journal of Neuroscience Methods.

[145]  A. J. Gandolfi,et al.  Use of Precision-Cut Liver Slices as an In Vitro Tool for Evaluating Liver Function , 1996, Toxicologic pathology.

[146]  S. Yamanaka,et al.  Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors , 2006, Cell.

[147]  T J Oberly,et al.  A comparison of the soft agar and microtitre methodologies for the L5178Y tk +/- mouse lymphoma assay. , 1997, Mutation research.

[148]  C. Larabell,et al.  Reversion of the Malignant Phenotype of Human Breast Cells in Three-Dimensional Culture and In Vivo by Integrin Blocking Antibodies , 1997, The Journal of cell biology.

[149]  Joshua F. Robinson,et al.  Discriminating classes of developmental toxicants using gene expression profiling in the embryonic stem cell test. , 2011, Toxicology letters.

[150]  A. Guillouzo,et al.  General review on in vitro hepatocyte models and their applications. , 2010, Methods in molecular biology.

[151]  Alexander Meissner,et al.  Induced pluripotent stem cells: current progress and potential for regenerative medicine. , 2009, Trends in molecular medicine.

[152]  Sindy K. Y. Tang,et al.  Multizone Paper Platform for 3D Cell Cultures , 2011, PloS one.

[153]  A. Fire,et al.  Specific interference by ingested dsRNA , 1998, Nature.

[154]  M. Negishi,et al.  Phenobarbital-elicited activation of nuclear receptor CAR in induction of cytochrome P450 genes. , 2000, Biochemical and biophysical research communications.

[155]  Patricia McGrath,et al.  A zebrafish assay for identifying neuroprotectants in vivo. , 2006, Neurotoxicology and teratology.

[156]  M. O'hare,et al.  Three-dimensional in vitro tissue culture models of breast cancer — a review , 2004, Breast Cancer Research and Treatment.

[157]  A. Piersma,et al.  Development of prediction models for three in vitro embryotoxicity tests in an ECVAM validation study. , 2000, In vitro & molecular toxicology.

[158]  Thomas B Knudsen,et al.  Zebrafish: as an integrative model for twenty-first century toxicity testing. , 2011, Birth defects research. Part C, Embryo today : reviews.

[159]  Anthony A. Hyman,et al.  Whither systems biology , 2011, Philosophical Transactions of the Royal Society B: Biological Sciences.

[160]  T. Sadler,et al.  Use of whole embryo culture for evaluating toxicity and teratogenicity. , 1984, Pharmacological reviews.

[161]  J. Goldstone,et al.  Cytochrome P450 1 genes in early deuterostomes (tunicates and sea urchins) and vertebrates (chicken and frog): origin and diversification of the CYP1 gene family. , 2007, Molecular biology and evolution.

[162]  C. Tyson,et al.  Evaluation of hepatocytes isolated by a nonperfusion technique in a prescreen for cytotoxicity. , 1980, Journal of toxicology and environmental health.

[163]  Kenneth M. Yamada,et al.  Taking Cell-Matrix Adhesions to the Third Dimension , 2001, Science.

[164]  Stefan Scholz,et al.  Developing a list of reference chemicals for testing alternatives to whole fish toxicity tests. , 2008, Aquatic toxicology.

[165]  A. J. Gandolfi,et al.  Precision-cut tissue slices: applications in pharmacology and toxicology. , 1995, Life sciences.

[166]  Shuo Lin,et al.  Chemical screening with zebrafish embryos. , 2011, Methods in molecular biology.

[167]  K Strange Model organisms: comparative physiology or just physiology? , 2000, American journal of physiology. Cell physiology.

[168]  T. Braunbeck,et al.  Is the fish embryo toxicity test (FET) with the zebrafish (Danio rerio) a potential alternative for the fish acute toxicity test? , 2009, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[169]  D. Jefferson,et al.  Culturing hepatocytes and other differentiated cells , 1984, Hepatology.

[170]  K. Reisinger,et al.  Identifying and characterizing chemical skin sensitizers without animal testing: Colipa's research and method development program. , 2010, Toxicology in vitro : an international journal published in association with BIBRA.

[171]  Judy Strickland,et al.  Bayesian integrated testing strategy (ITS) for skin sensitization potency assessment: a decision support system for quantitative weight of evidence and adaptive testing strategy , 2015, Archives of Toxicology.

[172]  Robert J Kavlock,et al.  Integration of dosimetry, exposure, and high-throughput screening data in chemical toxicity assessment. , 2012, Toxicological sciences : an official journal of the Society of Toxicology.

[173]  H. Moore,et al.  The regulation of self-renewal in human embryonic stem cells. , 2006, Stem cells and development.

[174]  E Dybing,et al.  Cytotoxic effects of N-acetyl-p-benzoquinone imine, a common arylating intermediate of paracetamol and N-hydroxyparacetamol. , 1984, Biochemical pharmacology.

[175]  Thomas Hartung,et al.  Integrated Testing Strategies (ITS) for safety assessment. , 2015, ALTEX.

[176]  D A New,et al.  WHOLE‐EMBRYO CULTURE AND THE STUDY OF MAMMALIAN EMBRYOS DURING ORGANOGENESIS , 1978, Biological reviews of the Cambridge Philosophical Society.

[177]  Nektarios Tavernarakis,et al.  Nemo: a computational tool for analyzing nematode locomotion , 2007, BMC Neuroscience.

[178]  H. Diekmann,et al.  A review of developmental toxicity screening using zebrafish larvae , 2011 .

[179]  C. Austin,et al.  Improving the Human Hazard Characterization of Chemicals: A Tox21 Update , 2013, Environmental health perspectives.

[180]  P. Olinga,et al.  Preparation and incubation of precision-cut liver and intestinal slices for application in drug metabolism and toxicity studies , 2010, Nature Protocols.

[181]  B P Schmid,et al.  Post-implantation embryo culture: validation with selected compounds for teratogenicity testing. , 1988, Xenobiotica; the fate of foreign compounds in biological systems.

[182]  Mehmet Fatih Yanik,et al.  Microfluidic system for on-chip high-throughput whole-animal sorting and screening at subcellular resolution , 2007, Proceedings of the National Academy of Sciences.

[183]  D. B. Myers,et al.  A replacement-first approach to toxicity testing is necessary to successfully reauthorize TSCA. , 2011, ALTEX.

[184]  J. Sulston,et al.  The embryonic cell lineage of the nematode Caenorhabditis elegans. , 1983, Developmental biology.

[185]  H. Babich,et al.  Acute cytotoxicities of polynuclear aromatic hydrocarbons determined in vitro with the human liver tumor cell line, HepG2 , 1988, Cell Biology and Toxicology.

[186]  Windy A. Boyd,et al.  A high-throughput method for assessing chemical toxicity using a Caenorhabditis elegans reproduction assay. , 2010, Toxicology and applied pharmacology.

[187]  Hiroshi Yamazaki,et al.  Evaluation of drug toxicity with hepatocytes cultured in a micro-space cell culture system. , 2011, Journal of bioscience and bioengineering.

[188]  Stefan Przyborski,et al.  Rat primary hepatocytes show enhanced performance and sensitivity to acetaminophen during three-dimensional culture on a polystyrene scaffold designed for routine use. , 2011, Assay and drug development technologies.

[189]  Paul C Melstrom,et al.  Reversible AChE inhibitors in C. elegans vs. rats, mice. , 2007, Biochemical and biophysical research communications.

[190]  Wim De Coen,et al.  Development of a screening assay to identify teratogenic and embryotoxic chemicals using the zebrafish embryo. , 2009, Reproductive toxicology.

[191]  S. Giri,et al.  Effects of bleomycin on 14C‐proline uptake, its incorporation into proteins and hydroxylation in collagenous proteins of hamster lung slices , 1985, Journal of applied toxicology : JAT.

[192]  Ivan Damjanov,et al.  Controlled synthesis of HBsAg in a differentiated human liver carcinoma-derived cell line , 1979, Nature.

[193]  Edward W Carney,et al.  Whole embryo culture: a "New" technique that enabled decades of mechanistic discoveries. , 2010, Birth defects research. Part B, Developmental and reproductive toxicology.

[194]  H. Sakaguchi,et al.  Development of an in vitro skin sensitization test using human cell lines: the human Cell Line Activation Test (h-CLAT). I. Optimization of the h-CLAT protocol. , 2006, Toxicology in vitro : an international journal published in association with BIBRA.

[195]  B. Nelson,et al.  Mechanical analysis of chorion softening in prehatching stages of zebrafish embryos , 2006, IEEE Transactions on NanoBioscience.

[196]  G Frank Gerberick,et al.  Chemical reactivity measurements: potential for characterization of respiratory chemical allergens. , 2011, Toxicology in vitro : an international journal published in association with BIBRA.

[197]  D. Clive,et al.  Cytogenetic distinction between the TK+ and TK- chromosomes in the L5178Y TK+/- 3.7.2C mouse-lymphoma cell line. , 1982, Mutation research.

[198]  Karen Blackburn,et al.  A framework for using structural, reactivity, metabolic and physicochemical similarity to evaluate the suitability of analogs for SAR-based toxicological assessments. , 2010, Regulatory toxicology and pharmacology : RTP.

[199]  Susanne Bremer,et al.  Validation of the Embryonic Stem Cell Test in the International ECVAM Validation Study on Three In Vitro Embryotoxicity Tests , 2004, Alternatives to laboratory animals : ATLA.

[200]  Paul A Clemons,et al.  The Connectivity Map: Using Gene-Expression Signatures to Connect Small Molecules, Genes, and Disease , 2006, Science.

[201]  A E McLean,et al.  Electron transport and protection of liver slices in the late stage of paracetamol injury of the liver. , 1989, British journal of pharmacology.

[202]  Jan Kaslin,et al.  Neurochemical and behavioural changes in zebrafish Danio rerio after systemic administration of 6‐hydroxydopamine and 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine , 2003, Journal of neurochemistry.

[203]  B. Quistorff,et al.  High zone-selectivity of cell permeabilization following digitonin-pulse perfusion of rat liver , 1989, Histochemistry.

[204]  Karen Augustine-Rauch,et al.  In vitro developmental toxicology assays: A review of the state of the science of rodent and zebrafish whole embryo culture and embryonic stem cell assays. , 2010, Birth defects research. Part C, Embryo today : reviews.

[205]  Gianfranco Piva,et al.  Carnosic acid from rosemary extracts: a potential chemoprotective agent against aflatoxin B1. An in vitro study , 2007, Journal of applied toxicology : JAT.

[206]  Andreas Bender,et al.  The challenges involved in modeling toxicity data in silico: a review. , 2012, Current pharmaceutical design.