Journal of Applied Toxicology

Cover image for Vol. 36 Issue 4

Early View (Online Version of Record published before inclusion in an issue)

Editor-in-Chief: Philip W. Harvey

Impact Factor: 2.982

ISI Journal Citation Reports © Ranking: 2014: 27/88 (Toxicology)

Online ISSN: 1099-1263


  1. 1 - 52
  1. Research articles

    1. Gene expression analyses of vitellogenin, choriogenin and estrogen receptor subtypes in the livers of male medaka (Oryzias latipes) exposed to equine estrogens

      Hiroshi Ishibashi, Masaya Uchida, Akiko Koyanagi, Yoshihiro Kagami, Teruhiko Kusano, Ayami Nakao, Ryoko Yamamoto, Nobuhiro Ichikawa, Nobuaki Tominaga, Yasuhiro Ishibashi and Koji Arizono

      Article first published online: 11 FEB 2016 | DOI: 10.1002/jat.3292

      No comprehensive data are yet available regarding the estrogenic potentials and risks of equine estrogens to aquatic organisms. We carried out expression analyses on estrogen-responsive genes in the livers of male medaka (Oryzias latipes) that were exposed to the equine estrogens for 3 days. Our qRT-PCR analyses revealed that the expression levels of hepatic estrogen-responsive genes in male medaka responded to various types and concentrations of equine estrogens. This is the first report describing the comprehensive analyses of in vivo estrogenicity of the equine estrogens in male medaka.

    2. Combined toxicity of heavy metal mixtures in liver cells

      Xialu Lin, Yuanliang Gu, Qi Zhou, Guochuan Mao, Baobo Zou and Jinshun Zhao

      Article first published online: 10 FEB 2016 | DOI: 10.1002/jat.3283

      Human exposure through air, water and food normally involves a mixture consisting of multiple metals. In this study, eight common heavy metals (Pb, Cd, Hg, Cu, Zn, Mn, Cr, Ni) that cause environmental contamination were selected to investigate the combined toxicity of different metal mixtures in HL7702 cells. Synergistic, antagonistic or additive effects of the toxicity were observed in different metal mixtures. These results suggest that the combined effects should be considered in the risk assessment of heavy metal co-exposure.

    3. Inhibitory effect of cadmium on estrogen signaling in zebrafish brain and protection by zinc

      Lina Chouchene, Elisabeth Pellegrini, Marie-Madeleine Gueguen, Nathalie Hinfray, François Brion, Benjamin Piccini, Olivier Kah, Khaled Saïd, Imed Messaoudi and Farzad Pakdel

      Article first published online: 9 FEB 2016 | DOI: 10.1002/jat.3285

      This study was conducted to assess the effects of Cd exposure on estrogen signaling in the zebrafish brain, and the potential protective role of Zn against Cd-induced toxicity. Effects on the transcriptional activation of estrogen receptors, aromatase B protein expression and molecular expression of related genes were examined. Our results demonstrate that Cd acts as a potent anti-estrogen in vivo and in vitro, and that Cd-induced estradiol antagonism can be reversed, at the protein level, by Zn supplement.

    4. Diurnal activity patterns as a sensitive behavioural outcome in fish: effect of short-term exposure to treated sewage and a sub-lethal PPCP mixture

      Steven D. Melvin, David R. Buck and Larelle D. Fabbro

      Article first published online: 9 FEB 2016 | DOI: 10.1002/jat.3284

      Diurnal activity patterns occur in many fish species and are sensitive to changes in natural factors. We exposed the mosquitofish (Gambusia holbrooki) to treated sewage and a mixture of Pharmaceuticals and Personal Care Products (PPCPs) identified in the wastewater, and quantified outcomes on diurnal rhythms. Daytime activity was abolished in male fish exposed to sewage and the mixture, whereas female activity was only significantly reduced with the PPCP mixture. Results demonstrate that diurnal activities provide a sensitive indication of sub-lethal toxicity in fish exposed to environmental pollutants.

    5. Integrated decision strategies for skin sensitization hazard

      Judy Strickland, Qingda Zang, Nicole Kleinstreuer, Michael Paris, David M. Lehmann, Neepa Choksi, Joanna Matheson, Abigail Jacobs, Anna Lowit, David Allen and Warren Casey

      Article first published online: 6 FEB 2016 | DOI: 10.1002/jat.3281

      The Interagency Coordinating Committee for the Validation of Alternative Methods (ICCVAM) evaluated a non-animal decision strategies to predict skin sensitization. Machine learning approaches integrated in vitro, in chemico and in silico data and six physicochemical properties for 120 substances to predict murine local lymph node assay (LLNA) outcomes. The seven models with the highest accuracy used a support vector machine with different combinations of predictor variables. The models outperformed individual non-animal methods and test batteries. This suggests that computational approaches are promising tools to effectively integrate data to identify potential skin sensitizers without animal testing.

    6. Gelucire and Gelucire-PEG400 formulations; tolerability in species used for non-clinical safety testing after oral (gavage) dosing

      Mikael Elander, Jette B. Boll, Anne S. Hojman and Allan D. Rasmussen

      Article first published online: 5 FEB 2016 | DOI: 10.1002/jat.3296

      A series of oral tolerability studies were conducted with Gelucire and Gelucire:PEG400 formulations in rats, dogs and minipigs in order to determine tolerable daily dose volumes in these species. It was concluded that Gelucire:PEG400 (90:10) was tolerated in Beagle dogs when administered at 1 ml kg–1 once daily for 39 weeks, and 100% Gelucire was tolerated in the rat and the minipig when administered once daily at 5 ml kg–1 for 5 days.

    7. Nanosuspension formulations of poorly water-soluble compounds for intravenous administration in exploratory toxicity studies: in vitro and in vivo evaluation

      Hisako Fujimura, Takao Komasaka, Taizo Tomari, Yasunori Kitano and Kouji Takekawa

      Article first published online: 5 FEB 2016 | DOI: 10.1002/jat.3280

      Nanosuspensions for intravenous injections were prepared using a mixer mill and zirconia beads with a vehicle of 2% (w/v) poloxamer 338, which was confirmed to induce no histamine release in dogs. Sterilized nanosuspensions were obtained by milling for 30 min, followed by autoclaving for 20 min at 121 °C and milling for 30 min (mill–autoclave–mill method). The enhancing effect of a nanosuspension on exposure in dogs and the versatility of the method were demonstrated.

    8. Depth-dependent stratum corneum permeability in human skin in vitro

      John Jay P. Cadavona, Hanjiang Zhu, Xiaoying Hui, Eui-Chang Jung and Howard I. Maibach

      Article first published online: 3 FEB 2016 | DOI: 10.1002/jat.3289

      The stratum corneum (SC) concentration–thickness profiles were determined for four model chemicals on intact and adhesive tape-stripped skin samples to clarify whether SC is a homogeneous barrier for chemical transport. Data analysis with the diffusion equation of Fick's second law permitted the chemical diffusion coefficient in SC. Results suggested the depth-dependency of SC permeability to panthenol, benzoic acid and butenafine; variation of the diffusion coefficient from the SC surface to the deeper layers agreed with a change in the diffusion coefficient over time in intact skin.

    9. Evaluation of kidney injury biomarkers in rat amniotic fluid after gestational exposure to cadmium

      Tania Jacobo-Estrada, Mariana Cardenas-Gonzalez, Mitzi Santoyo-Sánchez, Benjamín Parada-Cruz, Esther Uria-Galicia, Laura Arreola-Mendoza and Olivier Barbier

      Article first published online: 27 JAN 2016 | DOI: 10.1002/jat.3286

      Cadmium effects in fetal kidneys have not been widely studied, and no study has used early biomarkers for the detection of prenatal kidney injury. Pregnant Wistar rats were exposed to cadmium during gestation. Biomarkers of kidney injury were quantified in amniotic fluid (AF) samples, and fetal kidneys were processed for histological examination. Cadmium increased the levels of some biomarkers. Histological findings confirmed kidney alterations. In conclusion, kidney injury biomarkers in AF may be used to detect cadmium-induced fetal kidney damage.

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      Systems toxicology of chemically induced liver and kidney injuries: histopathology-associated gene co-expression modules

      Jerez A. Te, Mohamed Diwan M. AbdulHameed and Anders Wallqvist

      Article first published online: 4 JAN 2016 | DOI: 10.1002/jat.3278

      We proposed a protocol for selecting gene co-expression modules associated with chemical-induced injuries that classify 11 liver and eight kidney histopathology endpoints based on dose-dependent activation of the identified modules. We showed that the activation of the modules for a particular chemical exposure correlated with the severity of histopathological damage and could distinguish different types of organ-specific injuries. The generated modules provide a link between toxic chemical exposures, different molecular initiating events among underlying molecular pathways, and resultant organ damage.

  2. Review articles

    1. Metabolic activation and drug-induced liver injury: in vitro approaches for the safety risk assessment of new drugs

      M. José Gómez-Lechón, Laia Tolosa and M. Teresa Donato

      Article first published online: 22 DEC 2015 | DOI: 10.1002/jat.3277

      This review focuses on proposed in vitro screening strategies to predict and reduce idiosyncratic drug-induced liver injury associated with metabolic bioactivation. Metabolically competent biological systems (liver-derived cells, subcellular fractions), in combination with methods to reveal the formation of reactive intermediates (formation of adducts with proteins, metabolite trapping, enzyme inhibition assays), are commonly used to screen the reactivity of new molecules in early drug development. Several cell-based assays have also been proposed for the safety risk assessment of bioactivable compounds.

  3. Research articles

    1. Changes in RANKL and osteoprotegerin expression after chronic exposure to indoor air pollution as a result of cooking with biomass fuel

      Hirak Saha, Bidisha Mukherjee, Banani Bindhani and Manas Ranjan Ray

      Article first published online: 22 DEC 2015 | DOI: 10.1002/jat.3275

      The impact of indoor air pollution as a result of cooking with biomass on membrane-bound and serum receptor activator of nuclear factor-kappa ligand 1 (RANKL), its soluble decoy receptor osteoprotegerin (OPG) and osteoclast precursor CD14+CD16+ monocytes was investigated. Seventy-four pre-menopausal biomass-using women from eastern India and 65 control women who cooked with cleaner liquefied petroleum gas (LPG) were enrolled. Chronic exposure to biomass smoke increased membrane-bound and soluble RANKL and circulating osteoclast precursors but decreased OPG, suggesting an increased risk of bone resorption and consequent osteoporosis in their child-bearing age.

    2. Discrimination of skin sensitizers from non-sensitizers by interleukin-1α and interleukin-6 production on cultured human keratinocytes

      Daun Jung, Jeong-Hwan Che, Kyung-Min Lim, Young-Jin Chun, Yong Heo and Seung Hyeok Seok

      Article first published online: 22 DEC 2015 | DOI: 10.1002/jat.3274

      In vitro testing methods for classifying sensitizers could be valuable alternatives to in vivo sensitization testing using animal models, such as themurine local lymph node assay (LLNA) and the guinea pigmaximization test (GMT), but there remains a need for in vitro methods that are more accurate and simpler to distinguish skin sensitizers from non-sensitizers. Thus, the aim of our study was to establish an in vitro assay as a screening tool for detecting skin sensitizers using the human keratinocyte cell line, HaCaT.

    3. Long-term exposure to high levels of decabrominated diphenyl ether inhibits CD4 T-cell functions in C57Bl/6 mice

      Yan Feng, Weihong Zeng, Ying Wang, Hao Shen and Yan Wang

      Article first published online: 18 DEC 2015 | DOI: 10.1002/jat.3270

      Environmental exposure to decabrominated diphenyl ether (BDE-209) is ubiquitous and comparatively high burdens of BDE-209 have been found in occupationally environmental compartments and exposed personnel. In the present study, we observed impaired CD4 T-cell functions in mice exposed to a relatively high-dose of BDE-209 consecutively, accompanied by increased T regulatory cells in the blood. BDE-209 also suppressed the reactivity of CD4 T cells in vitro. Furthermore, antigen-specific CD4 T-cell responses to exogenous pathogens were prohibited in the exposed mice.

    4. You have full text access to this OnlineOpen article
      BMDExpress Data Viewer - a visualization tool to analyze BMDExpress datasets

      Byron Kuo, A. Francina Webster, Russell S. Thomas and Carole L. Yauk

      Article first published online: 15 DEC 2015 | DOI: 10.1002/jat.3265

      We developed BMDExpress Data Viewer, which contains two collections of tools, “Summary Visualization Tools” and “Dataset Exploratory Tools,” to visualize and analyze BMDExpress output files. Through two case studies, we demonstrate the capabilities of graphically examining transcriptomic dose–response datasets in a risk assessment context by comparing and observing trends in transcriptomic benchmark doses (BMDs) for biological processes and pathways. Our results illustrate that BMDExpress Data Viewer is a useful tool to visualize, explore and analyze BMDExpress output files.

    5. Sulfation of benzyl alcohol by the human cytosolic sulfotransferases (SULTs): a systematic analysis

      Lingtian Zhang, Katsuhisa Kurogi, Ming-Yih Liu, Alaina M. Schnapp, Frederick E. Williams, Yoichi Sakakibara, Masahito Suiko and Ming-Cheh Liu

      Article first published online: 11 DEC 2015 | DOI: 10.1002/jat.3268

      The aim of the present study was to identify human cytosolic sulfotransferases (SULTs) that are capable of sulfating benzyl alcohol and to examine the sulfation in cultured human cells and human organ homogenates. A systematic analysis revealed that four human SULTs, particularly SULT1A1, were capable of sulfating benzyl alcohol. HepG2 cells were able to generate sulfated benzyl alcohol under the metabolic settings. Moreover, the homogenates of human liver and small intestine displayed strong benzyl alcohol-sulfating activity.

    6. Development and application of a human PBPK model for bromodichloromethane to investigate the impacts of multi-route exposure

      Elaina M. Kenyon, Christopher Eklund, Teresa Leavens and Rex A. Pegram

      Article first published online: 9 DEC 2015 | DOI: 10.1002/jat.3269

      A refined human physiologically based pharmacokinetic (PBPK) model for bromodichloromethane (BDCM) (including new chemical-specific human parameters) was developed to evaluate the impact of BDCM exposure during showering and bathing on important measures of internal dose compared with oral exposure. Analyses demonstrated large contributions of dermal and inhalation exposure routes to an internal dose of the parent chemical reaching the systemic circulation. Thus, consideration of the contribution of multiple routes of exposure when evaluating risks from water-borne BDCM is highly desirable.

    7. Reconstituted high-density lipoprotein can elevate plasma alanine aminotransferase by transient depletion of hepatic cholesterol: role of the phospholipid component

      Eva Herzog, Ingo Pragst, Marcel Waelchli, Andreas Gille, Sabrina Schenk, Jochen Mueller-Cohrs, Svetlana Diditchenko, Paolo Zanoni, Marina Cuchel, Andreas Seubert, Daniel J. Rader and Samuel D. Wright

      Article first published online: 9 DEC 2015 | DOI: 10.1002/jat.3264

      Intravenous infusion of reconstituted high-density lipoprotein (HDL) causes simultaneous elevations in plasma cholesterol and alanine aminotransferase (ALT) in several species. The degree of cholesterol and ALT elevation correlates with the amount and type of phospholipid in the reconstituted HDL. Blocking hepatic cholesterol efflux by using either HDL pre-loaded with cholesterol or by genetic deletion of hepatic cholesterol transporter SR-BI, blocks rise of ALT. We conclude that ALT release is secondary to efflux of hepatic cholesterol into blood and can be overcome by reducing phospholipid content.

    8. Allopurinol induces innate immune responses through mitogen-activated protein kinase signaling pathways in HL-60 cells

      Akira Nakajima, Shingo Oda and Tsuyoshi Yokoi

      Article first published online: 7 DEC 2015 | DOI: 10.1002/jat.3272

      Allopurinol is a frequent cause of severe cutaneous adverse reactions (SCARs) in humans. Although SCARs have been suspected to be immune-mediated, the mechanisms of allopurinol-induced SCARs remain unclear. In this study, we demonstrate that treatment of HL-60 cells with allopurinol significantly increased the mRNA levels of pro-inflammatory cytokines, including IL-8, MCP-1, and TNFa, through activation of MAPK signaling pathways. These results indicate that innate immune responses induced by allopurinol might be involved in the development of allopurinol-induced SCARs.

    9. The minipig as a new model for the evaluation of doxorubicin-induced chronic toxicity

      Rosa Anna Manno, Andrea Grassetti, Germano Oberto, Abraham Nyska and Yuval Ramot

      Article first published online: 27 NOV 2015 | DOI: 10.1002/jat.3266

      Doxorubicin can cause life-threatening toxic effects in several organs, with cardiotoxicity being the major concern. Although a large number of animal models have been utilized to study doxorubicin toxicity, several restrictions limit their use. Since the Göttingen minipig is an accepted species for non-clinical safety assessment and translation to man, we aimed at exploring its use as a non-rodent animal model for safety assessment and regulatory toxicity studies using doxorubicin.

    10. Stratum corneum reservoir as a predictive method for in vitro percutaneous absorption

      Farhaan Hafeez, Audris Chiang, Xiaoying Hui, Hanjiang Zhu, Faraz Kamili and Howard I. Maibach

      Article first published online: 27 NOV 2015 | DOI: 10.1002/jat.3262

      Interaction between drug and stratum corneum (SC) is an important pharmacokinetic parameter in absorption. Previous in vivo studies showed the amount penetrating over a 96 h period could be predicted based on the linear relationship with the amount in SC after a 30 min application. We aimed to determine in vitro penetration behavior across SC by determining the association between quantity in SC 30 min after application and penetration after 24 h. Linear relationships are shown between quantity in SC 30 min after application and penetration after 24 h.

    11. Effect and mechanism of waterborne prolonged Zn exposure influencing hepatic lipid metabolism in javelin goby Synechogobius hasta

      Chao Huang, Zhi Luo, Christer Hogstrand, Feng Chen, Xi Shi, Qi-Liang Chen, Yu-Feng Song and Ya-Xiong Pan

      Article first published online: 25 NOV 2015 | DOI: 10.1002/jat.3261

      The present study was conducted to determine the effect and mechanism of waterborne Zn exposure influencing hepatic lipid deposition and metabolism in javelin goby Synechogobius hasta. S. hasta were exposed to four waterborne Zn concentrations (0.005, 0.18, 0.36 and 0.55 mg l-l, respectively) for 60 days. Sampling occurred at days 20, 40 and 60, respectively. Zn exposure increased Zn content, declined hepatic lipid content, and reduced lipogenic enzyme activities, including 6PGD, G6PD, ME and FAS. At days 20 and 60, Zn exposure decreased hepatic mRNA levels of 6PGD, G6PD, ME, FAS, ACCα, ACCβ, hormone-sensitive lipase (HSL)a, HSLb, SREBP-1, PPARα and PPARγ. However, the mRNA levels of CPT 1 and adipose triglyceride lipase increased following Zn exposure. On day 40, Zn exposure reduced hepatic mRNA expression of 6PGD, G6PD, ME, FAS, ACCα,ACCβ, HSLa, HSLb, SREBP-1 and PPARγ but increased mRNA expression of CPT 1, adipose triglyceride lipase and PPARα. For the first time, the present study provided evidence that chronic Zn exposure differentially influenced mRNA expression and activities of genes and enzymes involved in lipogenic and lipolytic metabolism in a duration-dependent manner, and provided new insight into the relationship between metal elements and lipid metabolism.

    12. Transcriptional and morphological effects of tamoxifen on the early development of zebrafish (Danio rerio)

      Liang Xia, Liang Zheng and Jun Liang Zhou

      Article first published online: 20 NOV 2015 | DOI: 10.1002/jat.3257

      Zebrafish embryos exposed to 500 µg l–1 of tamoxifen for 96 h caused a 20% reduction in heart rate and mild defects in caudal fin and skin. The expression of many endocrine-related genes was increased by tamoxifen exposure. The expression of metabolic-related genes and genes related to observed morphological changes were also upregulated by a high concentration of tamoxifen.

    13. Effects of soap–water wash on human epidermal penetration

      Hanjiang Zhu, Eui-Chang Jung, Christina Phuong, Xiaoying Hui and Howard Maibach

      Article first published online: 15 NOV 2015 | DOI: 10.1002/jat.3258

      Epidermal penetration was assayed with four chemicals to clarify effect of stratum corneum (SC) hydration on chemical penetration. Results showed accelerated penetration of benzoic acid and paraoxon quickly after surface wash, but reduced penetration of hydroquinone and benzoic acid 30 min post-decontamination. At the end of experiment, the lower hydroquinone penetration, greater paraoxon penetration and similar levels of benzoic acid and clonidine penetration resulted from surface wash. The observed wash-in effect agrees with the enhancement effect of SC hydration on the SC chemical absorption rate.

  4. Review articles

    1. An overview of the safety and biological effects of Bacillus thuringiensis Cry toxins in mammals

      Néstor Rubio-Infante and Leticia Moreno-Fierros

      Article first published online: 4 NOV 2015 | DOI: 10.1002/jat.3252

      Bacillus thuringiensis (Bt) Crystal proteins (Cry) are worldwide used bioinsecticides. This review summarizes and discusses current information regarding the biosafety and biological effects that Bt and its insecticidal Cry proteins elicit in mammals. Bt proteins are safe but are not innocuous to vertebrates. The effects of Cry proteins might be associated with immune-activating or allergic responses. More detailed studies are still needed on the effects of Cry toxins in mammals, as concerns regarding their possible health impact have been raised.

  5. Research articles

    1. Development of novel in vitro photosafety assays focused on the Keap1–Nrf2–ARE pathway

      Kyoko Tsujita-Inoue, Morihiko Hirota, Tomomi Atobe, Takao Ashikaga, Yoshiki Tokura and Hirokazu Kouzuki

      Article first published online: 28 OCT 2015 | DOI: 10.1002/jat.3234

      The immune response to photoallergens after UV-induced antigen formation is the same as in ordinary skin sensitization. We examined whether activation of the Keap1–Nrf2–ARE pathway, which is deeply involved in skin sensitization, could be used to assess the photoallergenic potential of chemicals using the reporter cell line AREc32 or KeratinoSensTM. Representative photoallergens activated this pathway after exposure to 5 J cm–2 UVA irradiation. The accuracy of predicting photoallergenicity/phototoxicity was 70% with AREc32 cells and 67% with KeratinoSensTM, and the specificity was 100%.

    2. You have full text access to this OnlineOpen article
      Predictive performance of the Vitrigel-eye irritancy test method using 118 chemicals

      Hiroyuki Yamaguchi, Hajime Kojima and Toshiaki Takezawa

      Article first published online: 15 OCT 2015 | DOI: 10.1002/jat.3254

      The sensitivity, specificity and accuracy of Vitrigel-EIT method in comparison to GHS were 90.1%, 65.9% and 80.5%, respectively. In case of eliminating nine chemicals showing pH 5 or lower, those were improved to 96.8%, 67.4% and 84.4%, respectively. Meanwhile, nine of 16 false-positive chemicals were classified irritant by EPA and immunohistologically confirmed to have an eye irritant potential. These data demonstrated that the Vitrigel-EIT method could provide excellent predictive performance to judge the widespread eye irritancy, including mild irritant chemicals.

    3. Benzoquinone toxicity is not prevented by sulforaphane in CD-1 mouse fetal liver cells

      Nicola A. Philbrook and Louise M. Winn

      Article first published online: 12 OCT 2015 | DOI: 10.1002/jat.3251

      Benzene is an environmental pollutant known to cause leukemia, however the mechanism of toxicity is unknown. We exposed cultured CD-1 mouse fetal liver cells to the benzene metabolite, benzoquinone, to determine its potential to cause DNA damage and alter DNA repair. Cells were also exposed to sulforaphane (SFN) to determine any potential protective effects against benzoquinone-mediated toxicity. Benzoquinone exposure led to a significant increase in ROS, DNA damage and decreased Ogg1 expression, which was not prevented by SFN.

    4. Comparison of outcomes obtained in murine local lymph node assays using CBA/J or CBA/Ca mice

      Yosuke Maeda, Haruka Hirosaki, Naoaki Yakata and Masahiro Takeyoshi

      Article first published online: 12 OCT 2015 | DOI: 10.1002/jat.3250

      Quantitative and qualitative comparisons of results in the murine local lymph node assay (LLNA) between the results obtained with CBA/J and CBA/Ca mice were made using five chemicals including typical contact sensitizers and non-sensitizers. Consequently, a significant difference was noted in disintegrations per minute (DPM) values per mouse derived from each strain of mice; however, no considerable difference was noted in the final outcomes, such as positive/negative decisions, stimulation index (SI) values and EC3 values in LLNA.

    5. Thyroid endocrine disruption of acetochlor on zebrafish (Danio rerio) larvae

      Mei Yang, Jingjin Hu, Shuying Li, Youning Ma, Wenjun Gui and Guonian Zhu

      Article first published online: 23 SEP 2015 | DOI: 10.1002/jat.3230

      Herbicide acetochlor has been suspected to disturb the thyroid endocrine system, but the underlying mechanisms have not yet been clarified. The present study suggested that zebrafish larvae in the time window could be used as a model for assessment of the thyroid endocrine disruption effects of pesticides. Acetochlor altered the mRNA expression of the hypothalamic–pituitary–thyroid axis-related genes and changed whole body thyroid hormone levels in zebrafish larvae, causing endocrine disruption of the thyroid system by simulating the biological activity of 3,5,3′-triiodothyronine.

    6. Functional expressions of adenosine triphosphate-binding cassette transporters during the development of zebrafish embryos and their effects on the detoxification of cadmium chloride and β-naphthoflavone

      Huancai Yin, Pengli Bai, Peng Miao, Mingli Chen, Jun Hu, Xudong Deng and Jian Yin

      Article first published online: 21 SEP 2015 | DOI: 10.1002/jat.3225

      This paper aimed to evaluate the physiological functions of adenosine triphosphate-binding cassette (ABC) transporters, Pgp (Abcb4), Mrp1, and Mrp2, at differential development stages of zebrafish embryos (4, 24, 48, and 72 hpf). The results indicated that both the gene expressions and activities of Pgp and Mrps increased with the development of embryos, which could cause an increasing tolerance of zebrafish embryos to cadmium chloride (CdCl2) and β-naphthoflavone (BNF).

    7. Oxidative stress-related DNA damage and homologous recombination repairing induced by N,N-dimethylformamide

      Cui Wang, Jinhuan Yang, Dezhao Lu, Yongsheng Fan, Meirong Zhao and Zhuoyu Li

      Article first published online: 21 SEP 2015 | DOI: 10.1002/jat.3226

      Our study demonstrated for the first time that DMF reduced the proliferation of human liver cells by causing oxidative DNA damage and double-strand breaks through the release of reactive oxygen species during biodegradation. Moreover, the ineffectiveness of the homologous repair pathway, leading to persistent DNA lesions, can partially explain the reason for DMF-related liver injury.

    8. Tris(2-chloroethyl)phosphate-induced cell growth arrest via attenuation of SIRT1-independent PI3K/Akt/mTOR pathway

      Wenjuan Zhang, Youjian Zhang, Zhiyuan Wang, Tian Xu, Cheng Huang, Wenjun Yin, Jing Wang, Wei Xiong, Wenhong Lu, Hongyan Zheng and Jing Yuan

      Article first published online: 17 SEP 2015 | DOI: 10.1002/jat.3223

      Tris(2-chloroethyl)phosphate (TCEP) as a flame retardant has been ubiquitously detected in the atmosphere, water and indoor dust samples as well as human milk and plasma samples. TCEP is classified as carcinogenic category 2 and toxic for reproduction category 1B. TCEP showed carcinogenicity in experimental animals in the liver and kidneys as well as cell loss in the brain. We investigated the in vitro effect of TCEP and TCEP-induced cell growth through the PI3K/Akt/mTOR pathway using the human non-tumor hepatic cell line L02 and human hepatoma cell line HepG2. We found that TCEP induced cell growth arrest associated with upregulated SIRT1 expression and inhibition of the PI3K/Akt/mTOR pathway; however, EX-527 aggravated cell growth arrest. The results indicated that TCEP induced cell growth arrest via attenuation of the SIRT1 independent PI3K/Akt/mTOR pathway.

    9. Benzo(a)pyrene inhibits migration and invasion of extravillous trophoblast HTR-8/SVneo cells via activation of the ERK and JNK pathway

      Liyuan Liu, Yingxiong Wang, Cha Shen, Junlin He, Xueqing Liu, Yubin Ding, Rufei Gao and Xuemei Chen

      Article first published online: 11 SEP 2015 | DOI: 10.1002/jat.3227

      The present study investigated the effect and mechanism of benzo(a)pyrene (BaP) on the invasion and migration of extravillous trophoblast HTR-8/SVneo cells. Treatment with BaP inhibited the migration and invasion of HTR-8/SVneo cells and changed the protein levels of MMP-2, MMP-9 and E-cadherin in cells. Moreover, BaP activated the MAPK signaling pathway. Pretreatment with specific inhibitors of MAPK rescued BaP-induced change in the migration and invasion of HTR-8/SVneo cells.

    10. Altered expression of histone deacetylases, inflammatory cytokines and contractile-associated factors in uterine myometrium of Long Evans rats gestationally exposed to benzo[a]pyrene

      Archana Laknaur, Terri-Lee Foster, Lesley E. Bobb, Aramandla Ramesh, Gwinnett M. Ladson, Darryl B. Hood, Ayman Al-Hendy and Chandrasekhar Thota

      Article first published online: 11 SEP 2015 | DOI: 10.1002/jat.3216

      There are no studies on the role of benzo[a]pyrene (BaP), an environmental toxicant, on preterm birth. In this study, pregnant rats treated with BaP delivered prematurely. Histopathology performed on the uterus, postpartum day 22, showed structural abnormalities in BaP-exposed rats. We observed a direct correlation between BaP metabolites and expression of interleukin-1β and -8, tumor necrosis factor α, connexin 43, cyclo-oxygenase-2, prostaglandin F receptor, histone deacetylase 5 (P < 0.05) and nuclear NFқBp65 and an inverse correlation with histone deacetylases 1 and 3 in the myometrium of BaP-exposed rats.

    11. The impact of caffeine on connexin expression in the embryonic chick cardiomyocyte micromass culture system

      Bhavesh K. Ahir and Margaret K. Pratten

      Article first published online: 24 AUG 2015 | DOI: 10.1002/jat.3219

      Cardiomyocytes are electrically coupled by gap junctions (GJs) defined as clusters of low-resistance multisubunit transmembrane channels composed of connexins (Cxs). The expression of Cx43, Cx45 and Cx40, which are present in cardiomyocytes, are known to be developmentally regulated. This study investigates the premise that alterations in GJs are one of the mechanisms by which teratogens may act. The most common human teratogen, caffeine was shown to affect cell-to-cell communication via the Cx43, Cx45 and Cx40, by alteration in expression and distribution in the embryonic chick cardiomyocytes MM culture system.

    12. Human ketosteroid receptors interact with hazardous phthalate plasticizers and their metabolites: an in silico study

      M. K. Sarath Josh, S. Pradeep, K. S. Vijayalekshmy Amma, R. Sudha Devi, S. Balachandran, M. N. Sreejith and Sailas Benjamin

      Article first published online: 24 AUG 2015 | DOI: 10.1002/jat.3221

      Phthalates are ubiquitous environmental pollutants, known for their adverse health effects in test animals, and of late in humans. Molecular interactions of diphthalates, respective monophthalates, phthalic acid and the known endocrine disruptor, bisphenol A with human ketosteroid receptors were explored using Glide (Schrödinger); and their binding efficiencies were compared with that of the natural steroids. From the in silico evidences, most of these diphthalates and their monophthalates showed potentials for anti-steroidal activity by interacting with human ketosteroid receptors.

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

      Joshua A. Harrill, Debra Layko, Abraham Nyska, Renee R. Hukkanen, Rosa Anna Manno, Andrea Grassetti, Marie Lawson, Greg Martin, Robert A. Budinsky, J. Craig Rowlands and Russell S. Thomas

      Article first published online: 17 AUG 2015 | DOI: 10.1002/jat.3211

      The role of AHR in mediating pathological changes in the liver prior to tumor formation was investigated in a 4-week, repeated-dose study using adult female wild-type (WT) and AHR knockout (AHR-KO) rats treated with varying concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD: 0, 3, 22, 100, 300 and 1000 ng kg−1 day−1). Treatment-related increases in the severity of liver and thymus pathology as well as changes in serum chemistry parameters were observed in WT, but not AHR-KO rats. Dose-dependent accumulation of TCDD was observed primarily in the liver of WT rats and primarily in the adipose tissue of AHR-KO rats.

    14. Sensitive periods for 17β-estradiol exposure during immune system development in sea bass head kidney

      Frauke Seemann, Thomas Knigge, Aurélie Duflot, Sabine Marie, Stéphanie Olivier, Christophe Minier and Tiphaine Monsinjon

      Article first published online: 16 AUG 2015 | DOI: 10.1002/jat.3215

      The influence of 17β-estradiol (E2) on estrogen receptor and cytokine gene expression, leukocyte populations and phagocytic activity was assessed in the juvenile sea bass (Dicentrarchus labrax) head kidney during organ regionalization from 98 to 239 dph. E2 exposure, beginning at 90 dph resulted in indirect and delayed modifications of interleukin 1β and estrogen receptor α gene expression, which may affect B-lymphocyte proliferation. The E2 treatment of 120 dph fish led to an increase in estrogen receptor β2 and a decrease in transforming growth factor β1 gene expression, which coincided with decreased phagocytic activity of head kidney lymphocytes and monocytes/macrophages.

    15. An in vitro human skin test for assessing sensitization potential

      S. S. Ahmed, X. N. Wang, M. Fielding, A. Kerry, I. Dickinson, R. Munuswamy, I. Kimber and A. M. Dickinson

      Article first published online: 7 AUG 2015 | DOI: 10.1002/jat.3197

      Chemical sensitization resulting in allergy is an important health issue. Here we describe a human in-vitro skin explant test for sensitization hazard assessment as an alternative approach to using animal models. This method measures histological damage in human skin as a readout of the immune response induced by the test material. We tested 44 chemicals and compared results to the mouse local lymph node assay LLNA and published human sensitization data with a correlation coefficient of 0.90 and 0.91, respectively.

    16. Perfluorinated chemicals, PFOS and PFOA, enhance the estrogenic effects of 17β-estradiol in T47D human breast cancer cells

      Pacharapan Sonthithai, Tawit Suriyo, Apinya Thiantanawat, Piyajit Watcharasit, Mathuros Ruchirawat and Jutamaad Satayavivad

      Article first published online: 3 AUG 2015 | DOI: 10.1002/jat.3210

      PFOS and PFOA do not possess estrogenic activity in T47D hormone-dependent human breast cancer cells. Both compounds enhance the effects of 17β-estradiol (E2) on estrogen response element (ERE) activation, expression of estrogen-responsive pS2 gene, ERK1/2 activation and cell growth.

    17. Antimicrobial agent triclosan is a proton ionophore uncoupler of mitochondria in living rat and human mast cells and in primary human keratinocytes

      Lisa M. Weatherly, Juyoung Shim, Hina N. Hashmi, Rachel H. Kennedy, Samuel T. Hess and Julie A. Gosse

      Article first published online: 23 JUL 2015 | DOI: 10.1002/jat.3209

      Triclosan (TCS) is an antimicrobial agent and mast cells are ubiquitous players in physiological processes and in diseases. TCS disrupts adenosine triphosphate production in mast cells, mouse fibroblasts and primary human keratinocytes and increases the oxygen consumption rate. TCS-methyl (no ionizable proton) affects neither degranulation nor adenosine triphosphate production at non-cytotoxic doses, indicating the effects of TCS are due to its proton ionophore structure. TCS is a mitochondrial uncoupler, affecting numerous cell types and functions via this mechanism.

  6. Research Articles

    1. BDE-209 inhibits pluripotent genes expression and induces apoptosis in human embryonic stem cells

      Lili Du, Wen Sun, Huili Zhang and Dunjin Chen

      Article first published online: 23 JUL 2015 | DOI: 10.1002/jat.3195

      Decabromodiphenyl ether (BDE-209) has been detected in human serum, semen, placenta, cord blood and milk worldwide. In this study, human embryonic stem cell lines FY-hES-10 and FY-hES-26 were used to evaluate the potential effects and explore the toxification mechanisms using low-level BDE-209 exposure. BDE-209 exposure could decrease pluripotent genes expression via epigenetic regulation and induce apoptosis through reactive oxygen species (ROS) generation in human embryonic stem cells in vitro.

  7. Research articles

    1. Proposed human stratum corneum water domain in chemical absorption

      Hanjiang Zhu, Eui-Chang Jung, Xiaoying Hui and Howard Maibach

      Article first published online: 23 JUL 2015 | DOI: 10.1002/jat.3208

      To better understand mechanisms of percutaneous absorption and skin decontamination, 21 compounds were studied for their affinities to stratum corneum (SC) and its subunits using a facile method. Differences between chemical absorption to intact SC and total contribution of protein and lipid domains suggest the possibility and significance of a water domain. A longer lag time of absorption into intact SC than to delipidized SC or SC lipid suggests the water domain may delay chemical binding to protein and lipid domains.

    2. Automated swimming activity monitor for examining temporal patterns of toxicant effects on individual Daphnia magna

      Simon Bahrndorff, Thomas Yssing Michaelsen, Anne Jensen, Laurits Faarup Marcussen, Majken Elley Nielsen and Peter Roslev

      Article first published online: 21 JUL 2015 | DOI: 10.1002/jat.3212

      We have evaluated an automated monitor for recording the swimming activity of Daphnia magna to establish temporal patterns of toxicant and temperature effects. The sensitivity of the monitor was evaluated by exposing D. magna to K2Cr2O7 and 2,4-dichlorophenol. Significant effects of toxicant concentrations, exposure time and incubation temperatures were observed. The results demonstrated that the monitor is capable of detecting sublethal behavioural effects that are toxicant and temperature dependent and can serve as a high-throughput screening tool in toxicity testing.

    3. Protein profiles of cardiomyocyte differentiation in murine embryonic stem cells exposed to perfluorooctane sulfonate

      Ying-Ying Zhang, Lei-Lei Tang, Bei Zheng, Ren-Shan Ge and Dan-Yan Zhu

      Article first published online: 15 JUL 2015 | DOI: 10.1002/jat.3207

      PFOS was classified as a weak embryotoxic chemical by using embryonic stem cell test procedure and it could block cardiomyocyte differentiation. In total, 176 differential proteins were identified by quantitative proteomics analysis. The differential proteins were classified by GO and connectivity analysis. PFOS affected 32 signaling pathways and mostly influenced the metabolism pathways. Five differentially expressed proteins were confirmed by western blotting. These results revealed potential new targets of PFOS on the developmental cardiovascular system.

    4. Defensive and adverse energy-related molecular responses precede tris (1, 3-dichloro-2-propyl) phosphate cytotoxicity

      Jinkang Zhang, Timothy D. Williams, James K. Chipman and Mark R. Viant

      Article first published online: 15 JUL 2015 | DOI: 10.1002/jat.3194

      To investigate the potential adverse effects of human exposure to tris (1, 3-dichloro-2-propyl) phosphate (TDCIPP), the cytotoxicity of this flame retardant to HepG2/C3A and A549 cells was evaluated by CCK-8 assays. The molecular responses to TDCIPP exposure were investigated using transcriptomic and metabolomic approaches. Defensive responses (e.g. xenobiotic metabolism) as well as energy-related changes were observed, which preceded the cytotoxic effects of TDCIPP in HepG2/C3A cells.

    5. Effects of sulpiride and ethylene glycol monomethyl ether on endometrial carcinogenicity in Donryu rats

      Yoshikazu Taketa, Kaoru Inoue, Miwa Takahashi, Yohei Sakamoto, Gen Watanabe, Kazuyoshi Taya and Midori Yoshida

      Article first published online: 14 JUL 2015 | DOI: 10.1002/jat.3206

      Sulpiride and ethylene glycol monomethyl ether (EGME) stimulate prolactin (PRL) secretion. Here, the effects of PRL on endometrial carcinogenicity were evaluated in rats. Sulpiride (200 ppm) inhibited the uterine carcinogenesis whereas EGME (1250 ppm) did not. Sulpiride prevented the onset of persistent estrus and induced high PRL and progesterone (P4) serum levels. These results suggest that disruption of the estrous cycle with a decrease in estradiol-17β to P4 ratio may explain the inhibitory effects of sulpiride on uterine carcinogenesis.

    6. The comparative toxicity of a reduced, crude comfrey (Symphytum officinale) alkaloid extract and the pure, comfrey-derived pyrrolizidine alkaloids, lycopsamine and intermedine in chicks (Gallus gallus domesticus)

      Ammon W. Brown, Bryan L. Stegelmeier, Steven M. Colegate, Dale R. Gardner, Kip E. Panter, Edward L. Knoppel and Jeffery O. Hall

      Article first published online: 14 JUL 2015 | DOI: 10.1002/jat.3205

      Comfrey (Symphytum officinale), a commonly used herb, contains pro-toxic dehydropyrrolizidine alkaloids, including lycopsamine and intermedine, and has consequently been internationally regulated with respect to its use. To help further define the toxicity of S. officinale, male, California White chicks were used to compare the toxicity of a crude, reduced comfrey alkaloid extract to purified lycopsamine and intermedine. Based on clinical, serum biochemical, tissue adduct concentrations and histopathological analysis, the reduced comfrey extract was more toxic than either pure lycopsamine or intermedine. This suggests a cautionary note when estimates of herbal toxicity are based upon the observed toxicity of isolated toxins.

    7. Kupffer cell-mediated exacerbation of methimazole-induced acute liver injury in rats

      Sho Akai, Yasuaki Uematsu, Koichi Tsuneyama, Shingo Oda and Tsuyoshi Yokoi

      Article first published online: 14 JUL 2015 | DOI: 10.1002/jat.3202

      Methimazole (MTZ) is known to cause liver injury in humans. It has been demonstrated that MTZ-induced liver injury in Balb/c mice is accompanied by T helper 2 cytokine-mediated immune responses; however, there is little evidence for immune responses associated with MTZ-induced liver injury in rats. We found that Kupffer cell-mediated immune responses are crucial factors for the exacerbation of MTZ-induced liver injury in rats, indicating apparent species differences in the immune-mediated exacerbation of liver injury between mice and rats.

    8. Angiogenesis is repressed by ethanol exposure during chick embryonic development

      Guang Wang, Shan Zhong, Shi-yao Zhang, Zheng-lai Ma, Jian-long Chen, Wen-hui Lu, Xin Cheng, Manli Chuai, Kenneth Ka Ho Lee, Da-xiang Lu and Xuesong Yang

      Article first published online: 14 JUL 2015 | DOI: 10.1002/jat.3201

      In this study, we investigated the anti-angiogenic effect of ethanol on the YSM during chick embryogenesis. The anti-angiogenic effect of ethanol was found to be associated with the excess ROS production. Both ethanol and AAPH (a ROS inducer) could inhibit cell proliferation, enhance apoptosis and repress expression of angiogenesis-related genes. This further supports our proposal that excess ROS production was central to the anti-angiogenic effect of ethanol.

    9. An assay to determine the sensitive window of embryos to chemical exposure using Xenopus tropicalis

      Lingling Hu, Lijiao Wu, Yingang Xue, Jingmin Zhu and Huahong Shi

      Article first published online: 14 JUL 2015 | DOI: 10.1002/jat.3200

      The Xenopus tropicalis embryos showed great variations of malformation in response to nine tested compounds during four separate 12-h exposure periods. Based on the value of score of malformations, the most sensitive exposure period of embryos was significantly distinguished for eight compounds. A rapid and valid assay was proposed to determine the 12-h sensitive window of embryos to chemical exposure using X. tropicalis.


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