Journal of Applied Toxicology

Cover image for Vol. 36 Issue 6

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 - 74
  1. Research articles

    1. Lack of genotoxic mechanisms in early-stage furan-induced hepatocellular tumorigenesis in gpt delta rats

      Daisuke Hibi, Yu Yokoo, Yuta Suzuki, Yuji Ishii, Meilan Jin, Aki Kijima, Takehiko Nohmi, Akiyoshi Nishikawa and Takashi Umemura

      Article first published online: 3 MAY 2016 | DOI: 10.1002/jat.3331

      Furan is shown to contaminate various foods and hepatocarcinogen in rodents. To elucidate furan-induced hepatocarcinogenic mechanisms, comprehensive medium-term analysis was conducted using gpt delta rats treated with furan at carcinogenic doses for 13 weeks. The reporter gene mutant frequencies in the liver were significantly increased. But, the number and area of glutathione S-transferase placental form-positive foci, and cell proliferation activity were significantly increased. Thus, it is highly probable that cell proliferation, but not genotoxic mechanisms, contribute to early-stage furan-induced hepatocarcinogenesis.

    2. Development of the Larval Amphibian Growth and Development Assay: effects of chronic 4-tert-octylphenol or 17β-trenbolone exposure in Xenopus laevis from embryo to juvenile

      Jonathan T. Haselman, Patricia A. Kosian, Joseph J. Korte, Allen W. Olmstead, Taisen Iguchi, Rodney D. Johnson and Sigmund J. Degitz

      Article first published online: 3 MAY 2016 | DOI: 10.1002/jat.3330

      The Larval Amphibian Growth and Development Assay (LAGDA) is a globally harmonized test guideline designed to evaluate apical effects of chronic chemical exposure with emphasis on endocrine disruption. Xenoestrogen, 4-tert-octylphenol, and xenoandrogen, 17β-trenbolone, were tested to evaluate the performance of the assay. Chemical-specific effects on thyroid homeostasis and reproductive development were identified. Notably, Müllerian duct development was the most sensitive endpoint affected by both chemicals. The LAGDA performed adequately to evaluate apical effects of chronic exposure to two endocrine-active compounds.

    3. Characterization of three human cell line models for high-throughput neuronal cytotoxicity screening

      Zhi-Bin Tong, Helena Hogberg, David Kuo, Srilatha Sakamuru, Menghang Xia, Lena Smirnova, Thomas Hartung and David Gerhold

      Article first published online: 3 MAY 2016 | DOI: 10.1002/jat.3334

      Three human neuronal cell lines were evaluated as high throughput screening models for neuronal cytotoxicity: SH-SY5Y neuroblastoma cells, LUHMES conditionally-immortalized dopaminergic neurons, and Neural Stem Cells. After 7 days of differentiation, LUHMES expressed the highest levels of neuronal markers. Differentiated LUHMES cells exhibited greater cytotoxic sensitivity to most of 32 suspected or known neurotoxicants than differentiated SH-SY5Y or NSCs, and greater cytotoxic sensitivity to 11 compounds compared to undifferentiated LUHMES cells.

    4. Effects of in vitro exposure to butylparaben and di-(2 ethylhexyl) phthalate, alone or in combination, on ovarian function

      Marina T. Guerra, Hayley C. Furlong, Wilma G. Kempinas and Warren G. Foster

      Article first published online: 2 MAY 2016 | DOI: 10.1002/jat.3335

      We evaluated the effects of Di-(2-ethylhexyl) phthalate (DEHP) and Butylparaben (BP), alone and in combination, on isolated mouse preantral follicle and human granulosa cell (hGC) cultures to study direct effects on follicle growth and ovarian steroidogenesis. Our results suggest that DEHP and BP adversely affect steroidogenesis from the preanatral stage onward and the effects of these chemicals are both stage-dependent and modified by co-exposure.

    5. Regucalcin counteracts tert-butyl hydroperoxide and cadmium-induced oxidative stress in rat testis

      Sara Correia, Cátia V. Vaz, Ana M. S. Silva, José E. Cavaco and Sílvia Socorro

      Article first published online: 25 APR 2016 | DOI: 10.1002/jat.3333

      To analyze the role of regucalcin (RGN)-modulating testicular oxidative stress, seminiferous tubules (SeT) from transgenic rats overexpressing RGN (Tg-RGN) and wild-type (WT) controls were cultured ex vivo in the presence/absence of pro-oxidant stimuli, tert-butyl hydroperoxide and cadmium chloride. The SeT of Tg-RGN animals displayed lower levels of oxidative stress and increased antioxidant defenses, underpinned by diminished rates of apoptosis. These findings support the antioxidant role of RGN in spermatogenesis.

    6. Biodistribution of polyacrylic acid-coated iron oxide nanoparticles is associated with proinflammatory activation and liver toxicity

      Diana Couto, Marisa Freitas, Vera Marisa Costa, Renan Campos Chisté, Agostinho Almeida, M. Arturo Lopez-Quintela, José Rivas, Paulo Freitas, Paula Silva, Félix Carvalho and Eduarda Fernandes

      Article first published online: 22 APR 2016 | DOI: 10.1002/jat.3323

      Iron oxide nanoparticles (IONs) have physical and chemical properties that render them useful for several new biomedical applications. Still, so far, in vivo safety studies of IONs with coatings of biomedical interest are still scarce. The aimof this study, therefore, was to clarify the acute biological effects of polyacrylic acid (PAA)-coated IONs, by determining their biodistribution and their potential proinflammatory and toxic effects in CD-1 mice.

    7. Neuroglial alterations in the zebrafish brain exposed to cadmium chloride

      Antonio Monaco, Maria C. Grimaldi and Ida Ferrandino

      Article first published online: 15 APR 2016 | DOI: 10.1002/jat.3328

      The effects of cadmium chloride on zebrafish brain were analysed by detecting the GFAP expression and mRNA levels of mbp, mpz and plp1 in specimens exposed to cadmium for 2, 7 and 16 days. A significant decrease in GFAP protein was observed already after 2 days reaching 50% after 16 days. This reduction was observed around ventricle areas and at the pial surface. Alterations in myelin were observed with upregulation of mpz mRNA levels. These data confirm the neurotoxic action of cadmium.

    8. Impact of silver nanoparticles on marine diatom Skeletonema costatum

      Jun Huang, Jinping Cheng and Jun Yi

      Article first published online: 15 APR 2016 | DOI: 10.1002/jat.3325

      Exposure to AgNPs induces excess intracellular reactive oxygen species, reduces chlorophyll a content and cell viability, inhibits the conversion of light energy into photosynthetic electron transport, and down-regulate the expression of the genes of the photosystem II reaction center protein in the exposed marine diatom. Scanning electron microscopy observation reveals that AgNPs obstruct the light absorption of algae. This study implies AgNPs might have serious consequences on the succession of the phytoplankton communities and aquatic ecosystem equilibrium.

    9. Accounting for data variability, a key factor in in vivo/in vitro relationships: application to the skin sensitization potency (in vivo LLNA versus in vitro DPRA) example

      S. Dimitrov, A. Detroyer, C. Piroird, C. Gomes, J. Eilstein, T. Pauloin, C. Kuseva, H. Ivanova, I. Popova, Y. Karakolev, S. Ringeissen and O. Mekenyan

      Article first published online: 15 APR 2016 | DOI: 10.1002/jat.3318

      A general methodology to analyze and integrate data variability when rescaling from one test to another is proposed. The methodology is demonstrated through the case study of rescaling in vitro DPRA reactivity to the in vivo LLNA skin sensitization potency classifications. A comprehensive statistical analysis evaluating the reliability and variability of LLNA and DPRA data a concept of grey zones around the thresholds and confidence probability have been introduced. The presented methodology is not restricted to skin sensitization only.

    10. A 28-year observational study of urinary cadmium and β2-microglobulin concentrations in inhabitants in cadmium-polluted areas in Japan

      Hoang Duc Phuc, Teruhiko Kido, Ho Dung Manh, Le Thai Anh, Nguyen Thi Phuong Oanh, Rie Okamoto, Akie Ichimori, Kazuhiro Nogawa, Yasushi Suwazono and Hideaki Nakagawa

      Article first published online: 15 APR 2016 | DOI: 10.1002/jat.3327

      A follow-up study was conducted over 28 years (1986–2014) to clarify the relationship between urinary cadmium (Cd) and β2-microglobulin (β2-MG) concentration in 28 inhabitants who lived in a previously Cd-polluted region in Japan. Urinary Cd concentration decreased by nearly half in both sexes between 1986 and 2008. However, it increased significantly from 2008 to 2014 in males. Urinary β2-MG concentrations tended to increase during the study. Age is more strongly associated with urinary β2-MG concentration than recent Cd body burden.

    11. Airborne nanoparticles (PM0.1) induce autophagic cell death of human neuronal cells

      Yu-Mi Jeon and Mi-Young Lee

      Article first published online: 15 APR 2016 | DOI: 10.1002/jat.3324

      Exposure to airborne nanoparticles PM0.1 (<100 nm in diameter) resulted in cell viability decrease and oxidative DNA damage. PM0.1-induced autophagic cell death occurred with increase in the expression of autophagy-related proteins. Moreover, 14 proteins were significantly changed upon exposure to PM0.1. Taken together, PM0.1-induced oxidative stress via ROS generation might play a key role in autophagic cell death and differential protein expressions in SH-SY5Y cells.

    12. You have full text access to this OnlineOpen article
      MicroRNA profiles in a monkey testicular injury model induced by testicular hyperthermia

      Ken Sakurai, Kei Mikamoto, Makoto Shirai, Takuma Iguchi, Kazumi Ito, Wataru Takasaki and Kazuhiko Mori

      Article first published online: 12 APR 2016 | DOI: 10.1002/jat.3326

      In this study, we identified 4 testicular-specific miRNAs, miR-34c-5p, miR-202-5p, miR-449a, and miR-508-3p based on next-generation sequencing of miRNAs from representative organs obtained from naïve mature cynomolgus monkeys. Next, miRNAs were profiled in a model of testicular injury induced by testicular hyperthermia. Microarray and PCR analyses revealed down-regulation of miR-34c-5p in the testis, which is enriched in meiotic cells, reflecting decreased numbers of pachytene spermatocytes and spermatids by the treatment.

    13. Accessing the molecular interactions of phthalates and their primary metabolites with the human pregnane X receptor using in silico profiling

      M. K. Sarath Josh, S. Pradeep, Aparna K. Balan, M. N. Sreejith and Sailas Benjamin

      Article first published online: 12 APR 2016 | DOI: 10.1002/jat.3321

      This molecular docking study using Glide evaluates a variety of molecular interactions of 12 commonly used diphthalates (generally reported as hazardous) and respective monophthalates with the ligand binding domain of the human pregnane X receptor (hPXR). Diisodecyl phthalate showed more affinity towards hPXR; whereas diisononyl phthalate and di(2-ethyhexyl)phthalate, the predominant plasticizers found in a variety of plastics and allied products, showed moderate binding. A higher affinity of certain phthalates to hPXR than its natural ligands and BPA (a proven endocrine disruptor) is alarming.

    14. Pyrazinamide induced hepatic injury in rats through inhibiting the PPARα pathway

      Yun Zhang, Hongli Guo, Hozeifa M. Hassan, Ping-ping Ding, Yijing Su, Yuming Song, Tao Wang, Lixin Sun, Luyong Zhang and Zhenzhou Jiang

      Article first published online: 12 APR 2016 | DOI: 10.1002/jat.3319

      In the present study, we described the hepatotoxic effects of PZA and the role of PPARα and its target genes in the downstream pathway including L-Fabp, Lpl, Cpt-1b, Acaa1, Apo-A1 and Me1 in this process. We found PZA induced the liver lipid metabolismdisorder and PPARα expression was down-regulated which had a significant inverse correlation with liver injury degree.

    15. Polymatin A from Smallanthus macroscyphus leaves: A safe and promising antidiabetic compound

      Carolina Serra-Barcellona, Stella M Honoré, Wilfredo M Cabrera, Natalia C Habib, Susana B Genta and Sara S Sánchez

      Article first published online: 7 APR 2016 | DOI: 10.1002/jat.3312

      Smallanthus macroscyphus is an herb native to South America whose leaves are a source of anti-diabetic compounds, although complete information about their safe use is not available yet. This study was developed to evaluate the toxicity profile of both 10% decoction and the sesquiterpene lactone polymatin A from S. macroscyphus leaves through in vitro cytotoxicity assays and in vivo subchronic oral toxicity.

    16. Perfluorooctane sulfonate (PFOS) impairs the proliferation of C17.2 neural stem cells via the downregulation of GSK-3β/β-catenin signaling

      Xuan Dong, Jianbin Yang, Xiaoke Nie, Jing Xiao and Shengyang Jiang

      Article first published online: 28 MAR 2016 | DOI: 10.1002/jat.3320

      PFOS has been recently documented to cause significant adverse effects on nervous system. We herein reported that PFOS exposure led to the downregulation of GSK-3β/β-catenin in C17.2 neural stem cells at a dose range of 25-200 nM, which is correlated with impaired proliferation of C17.2 cells. Our findings may help gain a better insight into the molecular mechanisms underlying PFOS-induced neurological deficits.

    17. Arsenic inhibits mast cell degranulation via suppression of early tyrosine phosphorylation events

      Juyoung Shim, Rachel H. Kennedy, Lisa M. Weatherly, Lee M. Hutchinson, Jonathan H. Pelletier, Hina N. Hashmi, Kayla Blais, Alejandro Velez and Julie A. Gosse

      Article first published online: 28 MAR 2016 | DOI: 10.1002/jat.3300

      Arsenite inhibits IgE-mediated degranulation of RBL-2H3 mast cells. Mast cells serve vital roles in many physiological processes and diseases. Upon activation, mast cells secrete mediators from cytoplasmic granules, in degranulation. Here, we report that arsenic does not affect degranulation stimulated by Ca2+ ionophore A23187, thapsigargin, or the G-protein activator compound 48/80, all of which bypass early signaling events. However, arsenic inhibits the Ca2+ influx into antigen-activated mast cells. Syk and phosphoinositide 3-kinase enzyme-linked immunosorbent assay data show that arsenic inhibits early phosphorylation events.

    18. Danio rerio ABC transporter genes abcb3 and abcb7 play a protecting role against metal contamination

      Adélaïde Lerebours, Van Vinh To and Jean-Paul Bourdineaud

      Article first published online: 28 MAR 2016 | DOI: 10.1002/jat.3313

      The involvement of two zebrafish ABC transporters in cadmium, zinc and mercury detoxification was characterized in a metal hypersensitive mutant of Escherichia coli. The E. coli tolC mutant expressing ABCB3 or ABCB7 transporters exhibited higher survival ratios and lower metal accumulation under a metal exposure condition than the controls.

  2. Short communications

    1. Triclosan is a mitochondrial uncoupler in live zebrafish

      Juyoung Shim, Lisa M. Weatherly, Richard H. Luc, Maxwell T. Dorman, Andy Neilson, Ryan Ng, Carol H. Kim, Paul J. Millard and Julie A. Gosse

      Article first published online: 28 MAR 2016 | DOI: 10.1002/jat.3311

      Antimicrobial agent triclosan (TCS) is a mitochondrial uncoupler in multiple cell types. Here, we present data, obtained with a Seahorse Bioscience XFe 96 Extracellular Flux Analyzer, indicating that TCS is also a mitochondrial uncoupler in living zebrafish [24 hour post-fertilization (hpf); 1 embryo per well]. TCS increases the oxygen consumption rate and proton leak and decreases ATP-linked respiration and spare respiratory capacity without affecting survival or heartbeat rate. These data indicate, for the first time, that TCS is a mitochondrial uncoupler in vivo.

  3. Research articles

    1. The neurotoxicity of DE-71: effects on neural development and impairment of serotonergic signaling in zebrafish larvae

      Xianfeng Wang, Lihua Yang, Qiangwei Wang, Yongyong Guo, Na Li, Mei Ma and Bingsheng Zhou

      Article first published online: 22 MAR 2016 | DOI: 10.1002/jat.3322

      Polybrominated diphenyl ethers have been proven to damage the central nervous system and affect neurobehavior in animals, but the underlying mechanism is still not clear. In our study, it was suggested that DE-71 exposure could cause impairment of the neural development and secondary effects on serotonergic system, and these lesions might be involved in the altered locomotor behavior in zebrafish larvae.

    2. Let-7a modulates particulate matter (≤ 2.5 μm)-induced oxidative stress and injury in human airway epithelial cells by targeting arginase 2

      Lei Song, Dan Li, Yue Gu, Xiaoping Li and Liping Peng

      Article first published online: 15 MAR 2016 | DOI: 10.1002/jat.3309


      Let-7a level was decreased in B2B cells after particulate matter (≤ 2.5 μm; PM2.5) exposure. Overexpression of let-7a decreased intracellular reactive oxygen species levels and the percentage of apoptotic cells after PM2.5 exposure. Arginase 2 (ARG2) was identified as a direct target of let-7a. Knockdown of ARG2 remarkably reduced oxidative stress and cellular injury. Restoration of ARG2 abrogated the protective effects of let-7a against PM2.5-induced injury. These data suggested that let-7a modulated PM2.5-induced oxidative stress and injury in human airway epithelial cells by targeting ARG2. The let-7a/ARG2 axis could be a therapeutic target for PM2.5-induced airway epithelial injury.

    3. Different mechanisms of action of 2, 2’, 4, 4’-tetrabromodiphenyl ether (BDE-47) and its metabolites (5-OH-BDE-47 and 6-OH-BDE-47) on cell proliferation in OVCAR-3 ovarian cancer cells and MCF-7 breast cancer cells

      Anna Karpeta, Anna Maniecka and Ewa Łucja Gregoraszczuk

      Article first published online: 14 MAR 2016 | DOI: 10.1002/jat.3316

      In present study, we investigated the influence of BDE-47 and its metabolites on proliferation in OVCAR-3 ovarian and MCF-7 breast cancer cells by examining their effects on cell-cycle regulation, oestrogen receptors expression and activation of ERK1/2 and PKCα phosphorylation. The results clearly show a tissue-dependent mechanism of BDE-47 action. BDE-47 increased cell-cycle genes and proteins expression in OVCAR-3, but not in MCF-7 cells. Hydroxylated metabolites in both cell lines acted on ERs expression and ERK1/2 and PKCα phosphorylation.

    4. 2,3,7,8-tetrachlorodibenzo-p-dioxin exposure influence the expression of glutamate transporter GLT-1 in C6 glioma cells via the Ca2+/protein kinase C pathway

      Jianya Zhao, Yan Zhang, Jianmei Zhao, Cheng Wang, Jiamin Mao, Ting Li, Xiaoke Wang, Xiaoke Nie, Shengyang Jiang and Qiyun Wu

      Article first published online: 14 MAR 2016 | DOI: 10.1002/jat.3294

      In this study, we investigated the role of TCDD in regulating the expression of glutamate transporter GLT-1 in astrocytes. TCDD, at concentrations of 0.1–100 nM, had no significantly harmful effect on the viability of C6 glioma cells. However, the expression of GLT-1 in C6 glioma cells was downregulated in a dose-and time-dependent manner. TCDD also caused activation of protein kinase C (PKC), as TCDD induced translocation of the PKC from the cytoplasmor perinuclear to the membrane.

    5. Neuropathy target esterase in mouse whole blood as a biomarker of exposure to neuropathic organophosphorus compounds

      Galina F. Makhaeva, Elena V. Rudakova, Larisa V. Sigolaeva, Ilya N. Kurochkin and Rudy J. Richardson

      Article first published online: 11 MAR 2016 | DOI: 10.1002/jat.3305

      The adult hen is the standard animal model for testing organophosphorus (OP) compounds for organophosphorus compound-induced delayed neurotoxicity (OPIDN). Recently, we developed a mouse model for the biochemical assessment of the neuropathic potential of OP compounds based on brain neuropathy target esterase (NTE) and acetylcholinesterase (AChE) inhibition. The present work represents further development of the mouse model aimed at using whole blood NTE as a biomarker of exposure to neuropathic OP compounds and predicting OPIDN risk in susceptible species by comparing blood NTE and AChE inhibition.

    6. Silver nanoparticles induce pro-inflammatory gene expression and inflammasome activation in human monocytes

      A. Murphy, A. Casey, G. Byrne, G. Chambers and O. Howe

      Article first published online: 10 MAR 2016 | DOI: 10.1002/jat.3315

      As a result of the increasing incidence of silver nanoparticle (AgNP) exposure, it is vital to determine their effect on the immune response. This study clearly demonstrates proinflammatory cytokine up-regulation after AgNP exposure to both a THP-1 monocyte cell line and primary human monocytes and the synergistic effect of combined exposure to AgNP and LPS. In addition, the release of IL-1β by THP-1 cells indicates inflammasome involvement. These results demonstrate the important immunological influence of AgNP.

  4. Review articles

    1. Dose site reactions and related findings after vaccine administration in safety studies

      Paul Baldrick

      Article first published online: 10 MAR 2016 | DOI: 10.1002/jat.3314

      A review of 30 recently published and in-house repeat dose toxicity studies with a variety of vaccines was performed. Results indicated no unexpected findings indicating vaccine toxicity, but classic signs of enhanced acute and/or chronic inflammation at the dose site compared with that seen in injected control animals, often accompanied by changes in draining lymph nodes and the spleen (lymphoid hyperplasia and/or increased weight). Other associated signs of a response to vaccine dosing were altered clinical pathology parameters (commonly raised blood neutrophil count and altered globulin level).

  5. Research articles

    1. Phenotypic and biomarker evaluation of zebrafish larvae as an alternative model to predict mammalian hepatotoxicity

      Sandra Verstraelen, Bernard Peers, Walid Maho, Karen Hollanders, Sylvie Remy, Pascale Berckmans, Adrian Covaci and Hilda Witters

      Article first published online: 4 MAR 2016 | DOI: 10.1002/jat.3288

      Zebrafish phenotypic assays have shown promise to assess human hepatotoxicity, though scoring of liver morphology remains subjective and difficult to standardize. Liver toxicity in zebrafish larvae at 5 days was assessed using gene expression as the biomarker approach, complementary to phenotypic analysis and analytical data on compound uptake. This approach aimed to contribute to improved hepatotoxicity prediction, with the goal of identifying biomarker(s) as a step towards the development of transgenic models for prioritization.

    2. You have full text access to this OnlineOpen article
      Optimization of an air–liquid interface exposure system for assessing toxicity of airborne nanoparticles

      Siiri Latvala, Jonas Hedberg, Lennart Möller, Inger Odnevall Wallinder, Hanna L. Karlsson and Karine Elihn

      Article first published online: 3 MAR 2016 | DOI: 10.1002/jat.3304

      An electrostatic airliquid interface exposure system was developed to assess the effects of pulmonary exposure to airborne nanoparticles. An applied electrostatic field strength and aerosol flow rate can be adjusted for achieving different particle doses. The highest amount of deposited particles was 2.2 μg cm2. Silver nanoparticles were found to cause cytotoxicity after 2 h exposure to 0.45 and 0.74 μg(Ag+)/cm2.

    3. Evaluation of the effects of deltamethrin on the fetal rat testis

      Anne-Marie Saillenfait, Dieynaba Ndiaye, Jean-Philippe Sabaté, Flavien Denis, Guillaume Antoine, Alain Robert, Virginie Rouiller-Fabre and Delphine Moison

      Article first published online: 2 MAR 2016 | DOI: 10.1002/jat.3310

      Pregnant Sprague–Dawley rats were administered deltamethrin, at a broad range of doses (0.1–10 mg kg−1 day−1), or di-n-hexyl phthalate (250 mg kg−1 day−1), by gavage, from gestational day 13 to 19. Unlike di-n-hexyl phthalate, deltamethrin had no significant effect on ex vivo testosterone production by gestational day 19.5 fetal testis or on the expression of several genes and proteins involved in the steroid synthesis pathway (SRB1, StAR, P450scc, 3βHSD, P450c17 and/or 17βHSD), up to the maternal toxic dose of 10 mg kg−1 day−1.

    4. Early metabolomics changes in heart and plasma during chronic doxorubicin treatment in B6C3F1 mice

      Laura K. Schnackenberg, Lisa Pence, Vikrant Vijay, Carrie L. Moland, Nysia George, Zhijun Cao, Li-Rong Yu, James C. Fuscoe, Richard D. Beger and Varsha G. Desai

      Article first published online: 2 MAR 2016 | DOI: 10.1002/jat.3307

      Metabolomics analyses of plasma and heart tissue from male B6C3F1 mice dosed weekly with 3 mg kg−1 doxorubicin or saline for 2, 3, 4, 6 or 8 weeks (cumulative doses of 6, 9, 12, 18 and 24 mg kg−1, respectively) identified potential early stage injury biomarkers of doxorubicin-induced cardiotoxicity. Select metabolites were altered after the 6 mg kg−1 cumulative dose whereas myocardial injury and cardiac pathology were not noted until after the 18 and 24 mg kg−1 cumulative doses, respectively.

    5. Involvement of calcium/calmodulin-dependent protein kinase II in methamphetamine-induced neural damage

      Xufeng Chen, Jingjing Xing, Lei Jiang, Wenyi Qian, Yixin Wang, Hao Sun, Yu Wang, Hang Xiao, Jun Wang and Jinsong Zhang

      Article first published online: 29 FEB 2016 | DOI: 10.1002/jat.3301

      Mounting evidence shows that METH exposure contributes to neurotoxicity, especially for the monoaminergic neurons. However, only a few studies have tried to unravel the mechanisms involved in METH-induced non-monoaminergic neural damage. Therefore, in the present study, we tried to explore the mechanisms for METH-induced neural damage in cortical neurons. Our results showed that METH significantly increased intracellular [Ca2+]i and up-regulated calmodulin (CaM) expression and activated calmodulin-dependent protein kinase II (CaMK II). Noteworthily, METH-induced neural damage can be partially retarded by CaM antagonist W7, CaMK II blocker KN93 and L-type Ca2+ channel specific inhibitor nifedipine. Collectively, our results suggest that Ca2+-CaM-CaMKII is involved in METH-mediated neurotoxicity, and it might suggest a potential target for the development of therapeutic strategies for METH abuse.

    6. Oxidative stress and lung pathology following geogenic dust exposure

      M. Leetham, J. DeWitt, B. Buck, D. Goossens, Y. Teng, J. Pollard, B. McLaurin, R. Gerads and D. Keil

      Article first published online: 29 FEB 2016 | DOI: 10.1002/jat.3297

      This study was designed to evaluate markers of systemic oxidative stress and lung histopathology following subacute exposure to geogenic dust with varying heavy metal content collected from a natural setting prone to wind erosion and used heavily for off-road vehicle recreation.

    7. Development of a quantitative morphological assessment of toxicant-treated zebrafish larvae using brightfield imaging and high-content analysis

      Samantha Deal, John Wambaugh, Richard Judson, Shad Mosher, Nick Radio, Keith Houck and Stephanie Padilla

      Article first published online: 29 FEB 2016 | DOI: 10.1002/jat.3290

      We developed a Computational Malformation Index that provides an objective manner for rapid phenotypic brightfield assessment of individual zebrafish larva in a developmental assay. By combining the use of high-content imaging with a very abbreviated human visual assessment, each larva can be quickly assessed for both detailed features as well as in-life parameters. Further, stability of these high-content measurements and the index performance were verified using a test set of zebrafish treated with two reference chemicals.

    8. Low-dose benzo[a]pyrene aggravates allergic airway inflammation in mice

      Rie Yanagisawa, Eiko Koike, Tin-Tin Win-Shwe, Takamichi Ichinose and Hirohisa Takano

      Article first published online: 25 FEB 2016 | DOI: 10.1002/jat.3308

      We examined to determine the effects of low dose BaP intratracheal administration on allergic airway inflammation and mediastinal lymph node (MLN) cell activation/proliferation in male C3H/HeJ mice. This study revealed that real-world exposure levels of BaP can, at least in part, enhance allergic airway inflammation by facilitating Th2 responses and activating MLN cells. In addition, high dose of BaP may contribute in activating both Th1 and Th2 responses.

    9. Differential cytotoxicity of copper ferrite nanoparticles in different human cells

      Javed Ahmad, Hisham A. Alhadlaq, Aws Alshamsan, Maqsood A. Siddiqui, Quaiser Saquib, Shams T. Khan, Rizwan Wahab, Abdulaziz A. Al-Khedhairy, Javed Musarrat, Mohd Javed Akhtar and Maqusood Ahamed

      Article first published online: 25 FEB 2016 | DOI: 10.1002/jat.3299

      Copper ferrite nanoparticles (NPs) have the potential to be applied in biomedical fields such as cell labeling and hyperthermia. However, there is a lack of information concerning the toxicity of copper ferrite NPs. We explored the cytotoxic potential of copper ferrite NPs in human lung (A549) and liver (HepG2) cells. Copper ferrite NPs were crystalline and almost spherically shaped with an average diameter of 35nm. Copper ferrite NPs induced dose-dependent cytotoxicity in both types of cells, evident by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide and neutral red uptake assays.

    10. Neverland regulates embryonic moltings through the regulation of ecdysteroid synthesis in the water flea Daphnia magna, and may thus act as a target for chemical disruption of molting

      Eri Sumiya, Yukiko Ogino, Kenji Toyota, Hitoshi Miyakawa, Shinichi Miyagawa and Taisen Iguchi

      Article first published online: 22 FEB 2016 | DOI: 10.1002/jat.3306

      To understand the physiological roles of ecdysteroids in Daphnia magna embryos, we analyzed the genes for ecdysteroid synthesis or degradation. The innate ecdysteroids titer during the embryogenesis showed two peaks after which embryonic moltings occur. Expression of neverland1 and shade increased before the first peak. Knocking-down neverland1 or shade inhibited embryonic moltings. Neverland1-knowckdown embryos showed lowered ecdysteroids titer. In situ hybridization revealed neverland1 was expressed in embryonic gut epithelium suggesting the site for an initial step of ecdysteroid synthesis.

    11. Cytotoxicity of various chemicals and mycotoxins in fresh primary duck embryonic fibroblasts: a comparison to HepG2 cells

      Xi Chen, Rhonda Murdoch, Daniel J. Shafer, Kolapo M. Ajuwon and Todd J. Applegate

      Article first published online: 18 FEB 2016 | DOI: 10.1002/jat.3298

      To cost-effectively screen the overall toxicity of a sample, especially in the case of food and feed ingredient quality control, a sensitive bioassay is necessary. Cell viability of fresh primary duck embryonic fibroblasts (DEF) in response to various chemicals and mycotoxins were determined. Results suggest that primary DEF are more sensitive to cytotoxins and mycotoxins compared to HepG2, and has great potential as an effective tool for in vitro cytotoxicity assessment.

    12. Cannabis effects on driving longitudinal control with and without alcohol

      Rebecca L. Hartman, Timothy L. Brown, Gary Milavetz, Andrew Spurgin, Russell S. Pierce, David A. Gorelick, Gary Gaffney and Marilyn A. Huestis

      Article first published online: 18 FEB 2016 | DOI: 10.1002/jat.3295

      Cannabis’ effects with/without alcohol on driving longitudinal control were evaluated against blood ∆9-tetrahydrocannabinol (THC) and breath alcohol concentrations (BrAC) in 18 drivers at the National Advanced Driving Simulator, University of Iowa. THC was associated with decreased speed, increased driving time >10% below the speed limit, and increased following distance during headway maintenance; BrAC, with increased standard deviation (SD) speed and increased time >10% above the speed limit. Cannabis’ association with slower driving and greater headway may suggest awareness of impairment, attempt to compensate.

  6. Review articles

    1. Copper toxicology, oxidative stress and inflammation using zebrafish as experimental model

      Talita Carneiro Brandão Pereira, Maria Martha Campos and Maurício Reis Bogo

      Article first published online: 17 FEB 2016 | DOI: 10.1002/jat.3303

      Copper is an essential micronutrient but also, potentially toxic and in excess, triggers oxidative stress responses that activate inflammation. Copper exposure can be easily modeled in zebrafish; a consolidated model in toxicology with increasing interest in immunity-related research. Copper toxicological and more recently pro-inflammatory effects have been investigated in both larval and adult zebrafish with breakthrough findings. Here, we provide an overview of copper metabolism in health and disease and its effects on oxidative stress and inflammation responses in zebrafish models.

  7. Research articles

    1. n-butylparaben induces male reproductive disorders via regulation of estradiol and estrogen receptors

      Linyuan Zhang, Sijin Ding, Peihuan Qiao, Li Dong, Miao Yu, Chong Wang, Ming Zhang, Lixia Zhang, Yimin Li, Ning Tang and Bing Chang

      Article first published online: 17 FEB 2016 | DOI: 10.1002/jat.3291

      Exposure to exogenous hormones during fetal or neonatal life leads to adverse reproductive outcomes. In this study, we investigated the possible mechanisms of reproductive disorders induced by n-butylparaben in utero and lactation in rats, which indicated that altered synthesis and metabolism of T and E2, the expressions of ERα and epigenetic hypomethylation of ERα contributed to the reproductive disorders. It is the first time to elucidate the possible mechanisms of reproductive disorders induced by n-butylparaben as a xenoestrogenic chemical.

    2. Quantitative evaluation of local pulmonary distribution of TiO2 in rats following single or multiple intratracheal administrations of TiO2 nanoparticles using X-ray fluorescence microscopy

      Guihua Zhang, Naohide Shinohara, Hirokazu Kano, Hideki Senoh, Masaaki Suzuki, Takeshi Sasaki, Shoji Fukushima and Masashi Gamo

      Article first published online: 16 FEB 2016 | DOI: 10.1002/jat.3287

      We quantitatively evaluated the pulmonary microdistribution (per mesh: 100 μm × 100 μm) of TiO2 in lung sections from rats following one, two, three, or four doses of TiO2 NPs at a same total dosage of 10 mg kg−1 using X-ray fluorescence microscopy. The evidence suggests that multiple-dose administrations do not offer remarkable advantages over single-dose administration on the pulmonary NP microdistribution, although multiple-dose administrations may reduce the variations in TiO2 content for each lung lobe.

    3. 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.

    4. 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.

    5. 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.

    6. 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.

    7. 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.

    8. 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.

    9. 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.

    10. 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.

    11. You have full text access to this OnlineOpen article
      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.

    12. 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.

    13. 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.

    14. 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.

    15. 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.

    16. 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.

    17. 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.

    18. 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.

    19. 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.

    20. 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.

    21. 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.

    22. 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.

    23. 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.

    24. 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%.

    25. 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.

    26. 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.

    27. 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.

    28. 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).

    29. 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.

    30. 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.

    31. 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.

    32. 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.

    33. 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.

    34. 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.


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