Journal of Applied Toxicology

Cover image for Vol. 34 Issue 5

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

Editor-in-Chief: Philip W. Harvey

Impact Factor: 2.597

ISI Journal Citation Reports © Ranking: 2012: 37/85 (Toxicology)

Online ISSN: 1099-1263


  1. 1 - 90
  1. Research Articles

    1. The in Vitro estrogenic activities of triclosan and triclocarban

      Hongyu Huang, Guizhen Du, Wei Zhang, Jialei Hu, Di. Wu, Ling Song, Yankai Xia and Xinru Wang

      Article first published online: 16 APR 2014 | DOI: 10.1002/jat.3012

      In the present study, we found that triclosan (TCS) and triclocarban (TCC) could induce the luciferase activities in the ER reporter gene assay, stimulate the proliferation of MCF-7 cells and change the expression of ERα and pS2 in MCF-7 cells. Moreover, TCS and TCC could alter the expression of multiple microRNAs (mir-22, mir-206 and mir-193b) in MCF-7 cells, which would help to understand the mechanisms of their estrogenic effects on regulating the expression of ERα.

    2. Usefulness of urinary kidney injury molecule-1 (Kim-1) as a biomarker for cisplatin-induced sub-chronic kidney injury

      Ken-ichiro Nan-ya, Masatomo Kajihara, Natsuki Kojima and Masakuni Degawa

      Article first published online: 16 APR 2014 | DOI: 10.1002/jat.2999

      We explored biomarkers suitable for monitoring sub-chronic kidney injury using rat models of kidney injury induced by cisplatin (CDDP). CDDP was intravenously administered to rats at 1.5 mg kg–1 for 4 days at 24-h intervals and for up to 10 weeks at weekly intervals, and the changes in blood and urine components were examined together with the histopathological changes in renal tissues. The present findings indicate that urinary kidney injury molecule-1 (Kim-1) is the most useful biomarker for CDDP-induced rat sub-chronic kidney injury.

    3. Evaluation of developmental toxicity using undifferentiated human embryonic stem cells

      Eui-Man Jung, Yeo-ul Choi, Hong-Seok Kang, Hyun Yang, Eui-Ju Hong, Beum-Soo An, Jun-young Yang, Ki. Hwan Choi and Eui-Bae Jeung

      Article first published online: 16 APR 2014 | DOI: 10.1002/jat.3010

      An embryonic stem cell test (EST) has been developed to evaluate the potential of chemicals with an in vitro system. In the present study, novel methods to screen toxic chemicals during the developmental process were evaluated using undifferentiated human embryonic stem (hES) cells. The embryotoxic chemicals influence their response to pluripotent ES cell markers. Our microarray data may be used as indicators for assessment of new chemicals. We define the hES cell viability and embryonic-specific differentiation by chemicals.

    4. Serum enhanced cytokine responses of macrophages to silica and iron oxide particles and nanomaterials: a comparison of serum to lung lining fluid and albumin dispersions

      David M. Brown, Helinor Johnston, Eva Gubbins and Vicki Stone

      Article first published online: 16 APR 2014 | DOI: 10.1002/jat.2998

      The effect of amorphous silica and iron oxide particles and nanoparticles (NPs) dispersed in cell culture medium supplemented with bovine serum albumin (BSA), lung lining fluid (LLF) or serum were examined in terms of macrophage viability and cytokine production. Silica induced a dose dependent increase in LDH release with evidence of a size difference between particles in some treatments. These data suggest that serum is very potent at enhancing macrophage responses to silica and iron oxide NPs and particles.

    5. In vitro exposure of DE-71, a penta-PBDE mixture, on immune endpoints in bottlenose dolphins (Tursiops truncatus) and B6C3F1 mice

      Jena R. Wirth, Margie M. Peden-Adams, Natasha D. White, Gregory D. Bossart and Patricia A. Fair

      Article first published online: 7 APR 2014 | DOI: 10.1002/jat.3008

      Following in vitro exposure to environmentally relevant DE-71, a penta-PBDE commercial mixture, concentrations (0–50 µg ml–1) in dolphin peripheral blood leukocytes (PBLs) and adult female B6C3F1 mouse splenocytes, lymphocyte proliferation and natural killer cell activity was measured.  Risk assessment using a parallelogram approach indicated that dolphin PBLs exhibited similar results to correlative field studies and mice were generally more sensitive than dolphins. The parallelogram approach was predictive for the immune endpoints in this study.

    6. Impact of acute arsenic and cadmium exposure on the expression of two haeme oxygenase genes and other antioxidant markers in common carp (Cyprinus carpio)

      Zsanett Jancsó and Edit Hermesz

      Article first published online: 7 APR 2014 | DOI: 10.1002/jat.3000

      The response of an organism to exposure to toxic metals is in many cases brought about by changes at the level of gene expression. In this study, the heme oxygenases genes (ho-1 and ho-2) of common carp Cyprinus carpio were identified, and the changes in gene expressions were analyzed from the aspect of Cd and As accumulation. Both ho-1 and ho-2 are transcriptionally induced by Cd and As, but their inductions differ in time course, dose response and tissue specificity.

    7. Usefulness of optical coherence tomography to detect central serous chorioretinopathy in monkeys

      Hyun-Kyu Park, Woori Jo, Hyun-Ji Choi, Bongcheol Kim, Gilnam Lee, Jeongbeob Seo, Suk Young Cho, Choung-Soo Kim, Eun Kyung Choi, Jung Jin Hwang, Joo Yong Lee, Young Hee Yoon and Woo-Chan Son

      Article first published online: 28 MAR 2014 | DOI: 10.1002/jat.3009

      It is difficult to detect ocular toxicity owing to the lack of appropriate evaluation methods and application of imaging tools to preclinical studies provides more information. Optical coherence tomography (OCT) is a valuable imaging method to screen drug-induced central serous chorioretinopathy (CSC), and will be useful to detect ocular toxicity in preclinical trials. To our knowledge, this is the first study to demonstrate CSC using OCT in a monkey model.

    8. Involvement of immune- and inflammatory-related factors in flucloxacillin-induced liver injury in mice

      Shohei Takai, Satonori Higuchi, Azusa Yano, Koichi Tsuneyama, Tatsuki Fukami, Miki Nakajima and Tsuyoshi Yokoi

      Article first published online: 20 MAR 2014 | DOI: 10.1002/jat.3002

      Flucloxacillin (FLX), which is a β-lactam antibiotic of the penicillin class, rarely causes drug-induced liver injury. The purpose of this study was to elucidate the mechanisms of FLX-induced liver injury in vivo. Toll-like receptor 4 (TLR4) ligands and inflammatory factors were significantly increased in FLX-administered mice. IL-17-related transcriptional factors and cytokines were increased, and the administration of recombinant IL-17 resulted in an exacerbation of the FLX-induced liver injury.TLR4-associated-signal transduction and IL-17 may be involved in FLX-induced liver injury.

    9. Exposure to parabens at the concentration of maximal proliferative response increases migratory and invasive activity of human breast cancer cells in vitro

      Sugandha Khanna, Philip R. Dash and Philippa D. Darbre

      Article first published online: 20 MAR 2014 | DOI: 10.1002/jat.3003

      Alkyl esters of p–hydroxybenzoic acid (parabens) are used widely as preservatives in consumer products. They have been measured in human breast tissue and can stimulate proliferation of oestrogen-responsive human breast cancer cells. We report here for the first time that long-term exposure (20 ± 2 weeks) to concentrations of methylparaben, n-propylparaben or n-butylparaben which give a maximal proliferative response also increased migratory and invasive properties of human breast cancer cell lines (MCF-7, T-47-D and ZR-75-1) in vitro.

    10. Molecular biomarkers of phospholipidosis in rat blood and heart after amiodarone treatment

      Nicola Bocchini, Mery Giantin, Federica Crivellente, Serena Ferraresso, Ivo Faustinelli, Mauro Dacasto and Patrizia Cristofori

      Article first published online: 18 FEB 2014 | DOI: 10.1002/jat.2992

      A set of phospholipidosis (PLD) biomarkers was identified in the blood and hearts of rats treated with amiodarone (AMD) using DNA microarrays. Overall, 545 and 8218 genes were modulated by AMD in hearts and blood, respectively; furthermore, some genes implicated in cellular phospholipid accumulation showed similar alterations. The usefulness of seven candidate genes was screened and confirmed by quantitative Real-Time PCR analysis. Presented data underscore the importance of transcriptional profiling in preclinical studies. Blood may be a surrogate tissue of cardiomyocyte PLD.

  2. Review Articles

    1. Nitroaromatic compounds: Environmental toxicity, carcinogenicity, mutagenicity, therapy and mechanism

      Peter Kovacic and Ratnasamy Somanathan

      Article first published online: 16 FEB 2014 | DOI: 10.1002/jat.2980

      Vehicle pollution is an increasing problem in the industrial world. Aromatic nitro compounds comprise a significant portion of the threat. In this review, the class includes nitro derivatives of benzene, biphenyls, naphthalenes, benzanthrone and polycyclic aromatic hydrocarbons, plus nitroheteroaromatic compounds. The numerous toxic manifestations are discussed. An appreciable number of drugs incorporate the nitroaromatic structure. The mechanistic aspects of both toxicity and therapy are addressed in the context of a unifying mechanism involving electron transfer, reactive oxygen species, oxidative stress and antioxidants.

  3. Research Articles

    1. Preliminary safety evaluation of a taurocholate-conjugated low-molecular-weight heparin derivative (LHT7): a potent angiogenesis inhibitor

      Farzana Alam, Seung Woo Chung, Seung Rim Hwang, Ji-young Kim, Jooho Park, Hyun Tae Moon and Youngro Byun

      Article first published online: 16 FEB 2014 | DOI: 10.1002/jat.2995

      In our previous studies, taurocholic acid (TA)-conjugated low-molecular-weight heparin derivative (LHT7) showed promising anti-angiogenic effects. For the development of this drug, LHT7, evaluation of its biological safety with mechanism-related side effects were performed. From this study, no other side effects have been found except for LHT7's accumulation in the liver and kidney owing its affinity to vascular endothelial growth factor (VEGF) and its saturation with TA at high doses.

    2. Interaction of citrate-coated silver nanoparticles with earthworm coelomic fluid and related cytotoxicity in Eisenia andrei

      Jin Il Kwak, Woo-Mi Lee, Shin Woong Kim and Youn-Joo An

      Article first published online: 16 FEB 2014 | DOI: 10.1002/jat.2993

      The earthworm Eisenia andrei was exposed to citrate-coated silver nanoparticles (cAgNPs), and the interaction of cAgNPs with earthworm coelomic fluid (ECF), the cytotoxicity of cAgNPs in earthworm coelomocytes was assessed. The aggregation and dissolution of cAgNPs increased in ECF, which contains various electrolytes that alter the properties of nanoparticles, and their subsequent toxicity. Microscopic and dissolution studies demonstrated that the aggregation of cAgNPs rapidly increased, and readily dissolved in ECF. The bioavailability of cAgNPs to earthworms induced lysosomal cytotoxicity.

    3. Carbon-based nanomaterials accelerate arteriolar thrombus formation in the murine microcirculation independently of their shape

      Martin Holzer, Peter Bihari, Marc Praetner, Bernd Uhl, Christoph Reichel, Janos Fent, Minnamari Vippola, Susan Lakatos and Fritz Krombach

      Article first published online: 14 FEB 2014 | DOI: 10.1002/jat.2996

      The aim of this study was to compare the acute effects of two differently shaped CBNs, fiber-shaped single-walled carbon nanotubes (SWCNT) and spherical ultrafine carbon black (CB), on thrombus formation as well as on leukocyte–endothelial cell interactions and leukocyte transmigration in the murine microcirculation upon systemic administration in vivo. The findings from this study suggest that systemic administration of CBNs accelerates arteriolar thrombus formation independently of the CBNs' shape, but does not induce leukocyte–endothelial cell interactions or leukocyte transmigration.

  4. Review Articles

    1. Role of the modulation of CYP1A1 expression and activity in chemoprevention: A mini review

      S. Badal and R. Delgoda

      Article first published online: 14 FEB 2014 | DOI: 10.1002/jat.2968

      An extra-hepatic Cytochrome P450 (CYP) enzyme, CYP1A1 has been comprehensively investigated for its ability to activate both exogenous and endogenous compounds into their carcinogenic derivatives, which link it to cancer initiation and progression. This review considers the role of CYP1A1 in cancer and takes an in-depth look at a number of phytochemicals, plant extracts and a few synthetic compounds that have been investigated for their ability to inhibit its activity and thus be considered for chemopreventive worth.

  5. Research Articles

    1. Development and utilization of an ex vivo bromodeoxyuridine local lymph node assay protocol for assessing potential chemical sensitizers

      W. C. Williams, C. Copeland, E. Boykin, S. J. Quell and D. M. Lehmann

      Article first published online: 14 FEB 2014 | DOI: 10.1002/jat.2983

      The murine local lymph node assay (LLNA) is widely used to identify chemicals that may cause allergic contact dermatitis. We developed an ex vivo BrdU-labeling procedure eliminating the need for in vivo injections. The ex vivo BrdU LLNA accurately predicted the sensitization potential of known sensitizers and non-sensitizers. This ex vivo BrdU labeling method offers predictive capacity comparable to previously established LLNA protocols while eliminating animal injections and the use of radioisotope.

    2. Comparative cytotoxicity of nanosilver in human liver HepG2 and colon Caco2 cells in culture

      Saura C. Sahu, Jiwen Zheng, Lesley Graham, Lynn Chen, John Ihrie, Jeffrey J. Yourick and Robert L. Sprando

      Article first published online: 12 FEB 2014 | DOI: 10.1002/jat.2994

      This report evaluates two in vitro cell culture models, human liver HepG2 cells and colon Caco2 cells, for screening the potential cytotoxicity of silver nanoparticles. It compares the cytotoxic potential of 20-nm silver in these two cell types. The nanosilver is cytotoxic to both the cell types, HepG2 being more sensitive. However, both the cell types are excellent in vitro systems for screening cytotoxicity of nanosilver.

    3. Fish multigeneration test with preliminary short-term reproduction assay for estrone using Japanese medaka (Oryzias latipes)

      Ataru Nakamura, Ikumi Tamura, Hitomi Takanobu, Masumi Yamamuro, Taisen Iguchi and Norihisa Tatarazako

      Article first published online: 12 FEB 2014 | DOI: 10.1002/jat.2981

      The long-term adverse effects of estrone on aquatic wildlife is of great concern. We exposed Japanese medaka (Oryzias latipes) to estrone (5.74-91.4 ng l–1) for 27 weeks across three generations. We found a decline in egg production and fertility in the second generation exposed to 91.4 ng l–1, much lower than known concentrations in urban rivers. Furthermore, histopathological abnormalities observed in the third generation exposed to 47.1 ng l–1 suggest that estrone exerts severe effects on the third or later generations.

  6. Review Articles

    1. You have full text access to this OnlineOpen article
      Mode of action human relevance (species concordance) framework: Evolution of the Bradford Hill considerations and comparative analysis of weight of evidence

      M. E. (Bette) Meek, Christine M. Palermo, Ammie N. Bachman, Colin M. North and R. Jeffrey Lewis

      Article first published online: 10 FEB 2014 | DOI: 10.1002/jat.2984

      The potential value of the mode of action (MOA)/human relevance (species concordance) framework in considering relative weight of evidence (WOE) amongst different cases and hypothesized MOA(s) is explored based on the content of several published assessments. The comparison is based on evolved Bradford Hill considerations rank ordered to reflect their relative contribution to WOE determinations for MOA based on experience internationally.

  7. Research Articles

    1. You have full text access to this OnlineOpen article
      Microvesicle-associated microRNA expression is altered upon particulate matter exposure in healthy workers and in A549 cells

      Valentina Bollati, Laura Angelici, Giovanna Rizzo, Laura Pergoli, Federica Rota, Mirjam Hoxha, Francesco Nordio, Matteo Bonzini, Letizia Tarantini, Laura Cantone, Angela C. Pesatori, Pietro Apostoli, Andrea A. Baccarelli and Pier Alberto Bertazzi

      Article first published online: 7 FEB 2014 | DOI: 10.1002/jat.2987

      Cell-derived microvesicles (MVs) are found in plasma and may transfer signals between tissues. In this article, we report in-vivo and in-vitro studies demonstrating that Particulate Matter (PM) affects systemic MV signaling by inducing MV release from alveolar cells into plasma. In-vivo microRNA screening showed increased miR-128 level in plasma MVs after PM exposure. In-vitro experiments confirmed PM-induced release of miR-128 in MVs from A549 alveolar cells. Future studies are warranted to determine the roles of MVs in mediating PM effects.

    2. Lipopolysaccharide exposure augments isoniazide-induced liver injury

      Yijing Su, Yun Zhang, Mi. Chen, Zhenzhou Jiang, Lixin Sun, Tao Wang and Luyong Zhang

      Article first published online: 6 FEB 2014 | DOI: 10.1002/jat.2979

      The results of this study demonstrate that LPS-induced inflammation makes rats more susceptible to INH hepatotoxicity. INH/LPS coadministration resulted in damage on bile acid metabolism, and cholestasis is considered as the main pattern of INH/LPS induced hepatotoxicity. TNF-α may be a factor in the ability of LPS to enhance the hepatotoxicity of INH. These results may have implications in idiosyncratic research attempting to improve the safety of INH.

    3. Development of a multiparametric in vitro model of skin sensitization

      Muriel Guyard-Nicodème, Eloise Gerault, Marion Platteel, Olivier Peschard, Wilfried Veron, Philippe Mondon, Svinareff Pascal and Marc G. J. Feuilloley

      Article first published online: 3 FEB 2014 | DOI: 10.1002/jat.2986

      A new in vitro model of skin sensitizing potential based on the measure of 4 parameters, i.e. trans-cutaneous permeation (Factor A), haptenation (Factor B), sensitization cytokines production (Factor C) and acute toxicity (Factor D) was described. A total of 32 compounds regularly employed in cosmetics were tested in this model. The correlation with in vivo assays reached 81.2% and the safety of the test (risk of false negative) reached 96.8 %.

    4. Facilitating the use of non-standard in vivo studies in health risk assessment of chemicals: a proposal to improve evaluation criteria and reporting

      Anna Beronius, Linda Molander, Christina Rudén and Annika Hanberg

      Article first published online: 30 JAN 2014 | DOI: 10.1002/jat.2991

      The purpose of this work was to propose tools that facilitate the use of non-standard toxicity studies in health risk assessment of chemicals. Criteria and guidance for evaluation of study reliability and relevance were based on requirements and recommendations identified from OECD test guidelines. A checklist for reporting such studies was also proposed. Together, these resources provide an approach to include all relevant data that may fill information gaps and reduce scientific uncertainty in health risk assessment conclusions.

    5. Differential effect of methyl-, butyl- and propylparaben and 17β-estradiol on selected cell cycle and apoptosis gene and protein expression in MCF-7 breast cancer cells and MCF-10A non-malignant cells

      Anna Maria Wróbel and Ewa Łucja Gregoraszczuk

      Article first published online: 30 JAN 2014 | DOI: 10.1002/jat.2978

      Parabens are alkyl esters of p-hydroxybenzoic acid used as antimicrobial preservatives in pharmaceuticals, foods and cosmetics. The present study was undertaken to determine the effect of methyl, butyl- and propylparaben, and 17β-estradiol as positive control, on cell cycle and apoptotic gene and protein expression in MCF-7 breast cancer and MCF-10A breast epithelial cells. The present results show differences between parabens and 17β-estradiol mechanisms of action in MCF-7, while similar to that of 17β-estradiol in non-transformed MCF-10A cells.

    6. Characterization of the biochemical effects of naphthalene on the mouse respiratory system using NMR-based metabolomics

      Jia-Huei Hong, Wen-Ching Lee, Yu-Ming Hsu, Hao-Jan Liang, Cho-Hua Wan, Chung-Liang Chien and Ching-Yu Lin

      Article first published online: 30 JAN 2014 | DOI: 10.1002/jat.2970

      Naphthalene, a ubiquitous environmental pollutant, has been shown to cause bronchiolar epithelial necrosis in the mouse airway. Metabolic variations in the bronchoalveolar lavage fluid and the lung tissues of naphthalene-treated mice and controls were examined using NMR-based metabolomics to identify the toxic mechanism. The metabolic changes may be related to lipid peroxidation, disruptions of membrane components, and imbalanced energy supply, and these results may partially explain the loss of cell membrane integrity in the airway epithelial cells of naphthalene-treated mice.

    7. Effects of cylindrospermopsin on a common carp leucocyte cell line

      Anna Sieroslawska and Anna Rymuszka

      Article first published online: 30 JAN 2014 | DOI: 10.1002/jat.2990

      Cylindrospermopsin (CYN) is a cytotoxin produced by different cyanobacterial species. The aim of the study was to elucidate the impact of cylindrospermopsin on a common carp (Cyprinus carpio L.) leucocyte cell line (CLC). The toxin at 10 and 1 µg ml–1 occurred to be evidently cytotoxic whereas at 0.1 µg ml–1 there were no effects on cell viability or cell number; however, a strong reduction of bacteria uptake was detected. The obtained results indicate that cylindrospermopsin may interfere with the basic functions of fish phagocytic cells.

    8. Benchmark dose of cadmium concentration in rice for renal effects in a cadmium-polluted area in Japan

      Kazuhiro Nogawa, Teruhiko Kido, Muneko Nishijo, Hideaki Nakagawa and Yasushi Suwazono

      Article first published online: 30 JAN 2014 | DOI: 10.1002/jat.2982

      We applied an updated hybrid approach to estimate the benchmark doses (BMD) and their 95% lower confidence limits (BMDL) of cadmium concentration in rice for renal effects in humans. The BMDL of cadmium in rice for β2-microglobulin was 0.25 mg/kg in men and 0.24 mg/kg in women. These estimated BMDLs may contribute to further discussion on the health risk assessment of cadmium exposure especially in terms of the food standard of cadmium concentration in rice.

    9. The effect of diesel exhaust exposure on blood–brain barrier integrity and function in a murine model

      Sayeh Heidari Nejad, Ryusuke Takechi, Benjamin J. Mullins, Corey Giles, Alexander N. Larcombe, Dean Bertolatti, Krassi Rumchev, Satvinder Dhaliwal and John Mamo

      Article first published online: 30 JAN 2014 | DOI: 10.1002/jat.2985

      Epidemiological studies indicate that exposure to diesel exhaust (DE) is associated with vascular-based disorders. An in vivo murine model was utilized to determine the effect of DE inhalation on neurovascular integrity. The study demonstrated that exposure to DE compromises the integrity and function of cerebral capillary vessels. Heightened neurovascular inflammation as a consequence of sub-chronic DE exposure may contribute to risk for neurovascular based disorders.

    10. Absence of mature microRNAs inactivates the response of gene expression to carcinogenesis induced by N-ethyl-N-nitrosourea in mouse liver

      Yang Luan, Xinming Qi, Liang Xu, Jin Ren and Tao Chen

      Article first published online: 30 JAN 2014 | DOI: 10.1002/jat.2973

      This study evaluates the role of microRNAs in chemical tumorigenesis by evaluating genomic gene expression in microRNA knockout mice treated with carcinogen N-ethyl-N-nitrosourea. The results demonstrate that the mouse liver in the absence of mature microRNAs could not appropriately respond to the carcinogenic insults to suppress the carcinogenesis induced by the carcinogen. Many tumor-suppressor functions, especially BRCA1 and P53 signaling pathways, were lost due to the absence of mature microRNAs. Thus, microRNAs play a critical role in chemical carcinogenesis.

    11. Application of the SOS/umu test and high-content in vitro micronucleus test to determine genotoxicity and cytotoxicity of nine benzothiazoles

      Yan Ye, Jiang Weiwei, Li Na, Ma Mei, Rao Kaifeng and Wang Zijian

      Article first published online: 30 JAN 2014 | DOI: 10.1002/jat.2972

      Cytotoxicity of BTs in both bacterial Salmonella typhimurium TA1535/pSK1002, MGC-803 and A549 cells was observed in a dose-dependent manner, and activation and inactivation were found for specific BTs after metabolism. Six of nine BTs were found to be genotoxic to either bacteria or cells or both, and structure alerts including heterocyclic amine and hacceptor-path3-hacceptor may contribute to the underlying genotoxic mechanism for 2-Aminobenzothiazole and 2-Hydroxy-benzothiazole respectively.

    12. Establishment of a short-term, in vivo screening method for detecting chemicals with juvenile hormone activity using adult Daphnia magna

      Ryoko Abe, Haruna Watanabe, Masumi Yamamuro, Taisen Iguchi and Norihisa Tatarazako

      Article first published online: 30 JAN 2014 | DOI: 10.1002/jat.2989

      Juvenile hormone (JH) and JH agonists have been shown to induce male offspring production in Daphnia using OECD TG211. The critical period for JH action to induce male offspring is 7-8 h (only 1 h) before ovulation. For the purpose of the detection of JH-like chemicals, we improved and simplified TG211 Annex7 and enabled us to save time and effort. The validity of a new testing method was verified by the test of positive and negative control chemicals.

    13. Zinc sulfide nanoparticles selectively induce cytotoxic and genotoxic effects on leukemic cells: involvement of reactive oxygen species and tumor necrosis factor alpha

      Sandeep Kumar Dash, Totan Ghosh, Soumyabrata Roy, Sourav Chattopadhyay and Debasis Das

      Article first published online: 30 JAN 2014 | DOI: 10.1002/jat.2976

      The present study describes the anti-cancer efficacy of synthesized zinc sulphide nanoparticles (ZnS NPs) on KG-1A cell line. pH dependent dissolution study of ZnS NPs showed selective internalization activity for leukemic cells and killed the cells by induction of apoptosis. Pre-treatment with ROS and TNF-α inhibitor confirmed that these nanoparticles were able to kill leukemic cells by generating an excess amount of ROS and thereby initiated TNF-α mediated apoptosis pathway.

    14. A longitudinal study assessing lens thickness changes in the eye of the growing beagle using ultrasound scanning: relevance to age of dogs in regulatory toxicology studies

      Juliana Maynard, Angela Sykes, Helen Powell, Guy Healing, Marietta Scott, Andrew Holmes, Sally-Ann Ricketts, Jane Stewart and Stewart Davis

      Article first published online: 16 JAN 2014 | DOI: 10.1002/jat.2967

      The lens is formed in utero with new secondary lens fibres added as outer layers throughout life in a growth pattern characteristic of the species. This study examined the time course of beagle lens growth to better understand the optimal starting age of dogs for safety studies to support adult versus paediatric indications, and to assess the feasibility of non-invasively monitoring lens growth with high frequency ultrasound. Ultrasound scanning was performed in six female beagle dogs using the Vevo770.

    15. Further studies toward a mouse model for biochemical assessment of neuropathic potential of organophosphorus compounds

      Galina F. Makhaeva, Elena V. Rudakova, Nichole D. Hein, Olga G. Serebryakova, Nadezhda V. Kovaleva, Natalia P. Boltneva, John K. Fink and Rudy J. Richardson

      Article first published online: 7 JAN 2014 | DOI: 10.1002/jat.2977

      Although the mouse does not readily exhibit clinical signs of organophosphorus compound-induced delayed neurotoxicity (OPIDN), it displays axonal lesions and expresses brain acetylcholinesterase (AChE) and neuropathy target esterase (NTE). The present research was performed as a further test of the hypothesis that inhibition of mouse brain AChE and NTE could be used to assess OPIDN potential using mouse brain preparations in vitro or employing mouse brain assays following dosing of OP compounds in vivo. Our results support this hypothesis.

    16. Gene expression profile of brain regions reflecting aberrations in nervous system development targeting the process of neurite extension of rat offspring exposed developmentally to glycidol

      Hirotoshi Akane, Fumiyo Saito, Ayako Shiraki, Nobuya Imatanaka, Yumi Akahori, Megu Itahashi, Liyun Wang and Makoto Shibutani

      Article first published online: 7 JAN 2014 | DOI: 10.1002/jat.2971

      Glycidol has shown effects on hippocampal neurogenesis after developmental exposure to rats. In this study, pregnant rats were given drinking water containing glycidol, and offspring were subjected to region-specific microarray analysis in four different brain areas. Gene expression alterations suggesting effects on neuronal plasticity were observed in all brain areas at 1000 ppm. Among them, genes in the hippocampal dentate gyrus were downregulated at ≥ 300 ppm, suggesting that neurogenesis might be the sensitive target of this type of toxicity.

    17. Evaluation of electric arc furnace-processed steel slag for dermal corrosion, irritation, and sensitization from dermal contact

      Mina Suh, Matthew J. Troese, Debra A. Hall, Blair Yasso, John J. Yzenas and Deborah M. Proctor

      Article first published online: 7 JAN 2014 | DOI: 10.1002/jat.2974

      Electric arc furnace (EAF) steel slag is alkaline and contains metals and thus has potential to cause dermal effects. To assess dermal irritation, corrosion, and sensitizing potential of EAF slag, in vitro and in vivo dermal toxicity assays were conducted. EAF slag was not corrosive, irritating, or sensitizing in any tests. These findings are supported by the observation that metals in EAF slag occur as oxides of low solubility with leachates that are well below toxicity characteristic leaching procedure limits.

    18. The immunosuppressant tributyltin oxide blocks the mTOR pathway, like rapamycin, albeit by a different mechanism

      Ahmed M. Osman and Henk van Loveren

      Article first published online: 26 DEC 2013 | DOI: 10.1002/jat.2966

      We report here that tributyltin oxide (TBTO) inhibits, like rapamycin, the p70S6k1 pathway, though by a different mechanism in EL4 cell line. Both TBTO and rapamycin downregulated the phosphorylation state of S6k1 assessed at Ser421/Thr424, but in contrast to rapamycin, TBTO decreased the total S6k1 protein. In addition, TBTO reduced the levels of RPS6 (Ser236) and eIF4B (Ser422), downstream substrates of S6k1. Further, TBTO and rapamycin differentially affected 4E-binding protein 1, a repressor of the cap-binding protein eIF4E.

    19. Use of the mouse ear vesicant model to evaluate the effectiveness of ebselen as a countermeasure to the nitrogen mustard mechlorethamine

      Anju Lulla, Sandra Reznik, Louis Trombetta and Blase Billack

      Article first published online: 17 DEC 2013 | DOI: 10.1002/jat.2969

      The goal of the present study was to demonstrate for the first time the effectiveness of ebselen (EB-1) as a vesicant countermeasure in an in vivo model. Using the mouse ear vesicant model (MEVM), HN2, applied topically, showed a dose-dependent injury. To examine HN2 countermeasure activity, either hydrocortisone (HC) or EB-1 was administered as three topical treatments after HN2 exposure. Both HC and EB-1 were found to reduce tissue swelling associated with HN2 toxicity 24 h after vesicant exposure.

    20. Identification of differentially expressed proteins related to organophosphorus-induced delayed neuropathy in the brains of hens

      Ying Jiang, Xiaohui Liu, Shuangyue Li, Yanning Zhang, Fengyuan Piao and Xiance Sun

      Article first published online: 12 DEC 2013 | DOI: 10.1002/jat.2965

      In this study, eight differentially expressed proteins related to organophosphate-induced delayed neuropathy (OPIDN) were identified in the brains of hens on day 5. Among these proteins, downregulated expression of glutamine synthetase (GS) was further confirmed. Moreover, TOCP exposure increased the levels of glutamate and [Ca2+]i, and decreased the level of glutamine, which indicates that the downregulated expression of GS might be associated with OPIDN through the disruption of homeostasis of the glutamate–glutamine cycle and [Ca2+]i in the early stage after TOCP administration.

    21. Proteomic analysis of perfluorooctane sulfonate-induced apoptosis in human hepatic cells using the iTRAQ technique

      Qingyu Huang, Jie Zhang, Siyuan Peng, Miaomiao Du, Sawyen Ow, Hai Pu, Chensong Pan and Heqing Shen

      Article first published online: 3 DEC 2013 | DOI: 10.1002/jat.2963

      Environmental perfluorooctane sulfonate (PFOS) exposure has been associated with a number of adverse health effects. However, the mechanisms associated with PFOS toxicity remain unclear. In the current proteomic study, 18 differential proteins with dose-dependent changes were identified in PFOS-exposed L-02 cells by iTRAQ-labeling and nanoLC-MS/MS. Furthermore, induction of apoptosis was proposed as a key mechanism for PFOS cytotoxicity. These results may provide useful insights into the mechanisms of PFOS-mediated hepatotoxicity and assist in the development of biomarkers indicative of PFOS exposure.

    22. Increased methylmercury toxicity related to obesity in diabetic KK-Ay mice

      Megumi Yamamoto, Rie Yanagisawa, Eriko Motomura, Masaaki Nakamura, Mineshi Sakamoto, Motohiro Takeya and Komyo Eto

      Article first published online: 15 NOV 2013 | DOI: 10.1002/jat.2954

      Greater methylmercury (MeHg) toxicity was observed in KK-Ay type 2 diabetic mice compared with that of C57BL/6J mice at the same dose of MeHg per body weight. MeHg-treated KK-Ay mice showed hind-limb clasping in the final stage of the experiment. Body fat gain and low mercury accumulation in adipose tissue increased MeHg concentrations in organs and enhanced MeHg toxicity. Tissue injuries in MeHg-exposed KK-Ay mice were detected by CD204 expression.

    23. Circulating microRNA 122 in the methionine- and choline-deficient mouse model of non-alcoholic steatohepatitis

      John D. Clarke, Tatiana Sharapova, April D. Lake, Eric Blomme, Jonathan Maher and Nathan J. Cherrington

      Article first published online: 12 NOV 2013 | DOI: 10.1002/jat.2960

      The methionine choline-deficient diet (MCD) is a frequently used hepatotoxicity animal model of non-alcoholic steatohepatitis (NASH) that induces hepatic transaminase (ALT, AST) elevations and hepatobiliary histological changes similar to those observed in human NASH. Liver-specific microRNA-122 (miR-122) has recently been proposed as a sensitive and specific circulating biomarker of hepatic injury. This study shows that miR-122 serum levels are elevated earlier than transaminases and may be useful as a sensitive biomarker for the early detection of NAFLD-associated liver injury.

    24. Deltamethrin induced an apoptogenic signalling pathway in murine thymocytes : exploring the molecular mechanism

      Anoop Kumar, D. Sasmal and Neelima Sharma

      Article first published online: 12 NOV 2013 | DOI: 10.1002/jat.2948

      The immunotoxic effects of DLM on mammalian system and its mechanism is still unclear. This study has been designed to first observe the binding affinity of DLM to immune cells and its effects on immune system. The results imply that DLM has a strong binding affinity towards the CD4 and CD8 receptors. DLM induces apoptosis in murine thymocytes in a concentration-dependent manner. Caspase, ROS and GSH act as critical mediators in DLM-induced apoptosis. The posibility of caspase-independent pathways also exists.

    25. Combined antitumor effects of bee venom and cisplatin on human cervical and laryngeal carcinoma cells and their drug resistant sublines

      Goran Gajski, Tamara Čimbora-Zovko, Sanjica Rak, Marko Rožman, Maja Osmak and Vera Garaj-Vrhovac

      Article first published online: 12 NOV 2013 | DOI: 10.1002/jat.2959

      The present study investigated the possible combined anticancer ability of bee venom (BV) and cisplatin towards two pairs of tumour cell lines: parental cervical carcinoma HeLa cells and their cisplatin-resistant HeLa CK subline, as well as laryngeal carcinoma HEp-2 cells and their cisplatin-resistant CK2 subline. Our results suggest that combined treatment with BV could be useful from the point of minimizing the cisplatin concentration during chemotherapy, consequently reducing and/or postponing the development of cisplatin resistance.

  8. Review Articles

    1. Toxic compounds in honey

      Md. Nazmul Islam, Md. Ibrahim Khalil, Md. Asiful Islam and Siew Hua Gan

      Article first published online: 11 NOV 2013 | DOI: 10.1002/jat.2952

      There is a wealth of information about the nutritional and medicinal properties of honey. However, honey may contain compounds that may lead to toxicity. A compound not naturally present in honey, named 5-hydroxymethylfurfural (HMF), may be formed during the heating or preservation processes of honey. HMF has gained much interest, as it is commonly detected in honey samples, especially samples that have been stored for a long time. HMF is a compound that may be mutagenic, carcinogenic and cytotoxic.

  9. Research Articles

    1. Hepatic fibrogenesis and transforming growth factor/Smad signaling activation in rats chronically exposed to low doses of lead

      Rossana C. Perez Aguilar, Stella M. Honoré, Susana B. Genta and Sara S. Sánchez

      Article first published online: 6 NOV 2013 | DOI: 10.1002/jat.2955

      The present work describes the role of lead in promoting liver fibrogenesis and suggests mechanisms in the underlying process. Our results report for the first time, that chronic exposure to low doses of lead could trigger the phenotypic transformation of hepatic stellate cells, secretion of the extracelular matrix and upregulated expression of TGFβ1/Smad signaling. Taken together, these results suggest a role of lead in promoting a fibrogenic state with altered hepatic structure.

    2. Calcium montmorillonite clay reduces AFB1 and FB1 biomarkers in rats exposed to single and co-exposures of aflatoxin and fumonisin

      Nicole J. Mitchell, Kathy S. Xue, Shuhan Lin, Alicia Marroquin-Cardona, Kristal A. Brown, Sarah E. Elmore, Lili Tang, Amelia Romoser, Wentzel C. A. Gelderblom, Jia-Sheng Wang and Timothy D. Phillips

      Article first published online: 6 NOV 2013 | DOI: 10.1002/jat.2942

      The binding capacity of UPSN, a calcium montmorillonite clay, was evaluated after aflatoxin (AF) and fumonisin (FB) single and co-exposures in Fischer 344 rats. UPSN decreased AFM1 and FB1 urinary biomarker excretion by 88–97% and 45–85%, respectively. When in combination, both AFM1 and FB1 sorption occurred, but capacity was decreased by half. In the absence of UPSN, combined AFB1 and FB1 treatment decreased corresponding urinary biomarkers by 67% and 45%, respectively, but increased levels of AFB1-albumin adduct compared with AFB1-treated rats.

  10. Review Articles

    1. Drug safety testing paradigm, current progress and future challenges: an overview

      Varun Ahuja and Sharad Sharma

      Article first published online: 29 OCT 2013 | DOI: 10.1002/jat.2935

      Early assessment of the safety of new molecules in the pharmaceutical industry is a multi-dimensional task involving predictive systems and screening approaches to aid in the optimization of lead compounds prior to their entry into the development phase. This study discusses the developments that allow toxicologists to anticipate safety problems and plan ways to address them, earlier than ever before. Today's toxicologist should collaborate with medicinal chemists, pharmacologists, and clinicians and these value-adding contributions will change traditional toxicologists from side-effect identifiers to drug development enablers.

  11. Research Articles

    1. Protective effects of schizandrin and schizandrin B towards cisplatin nephrotoxicity in vitro

      Valérian Bunel, Marie-Hélène Antoine, Joëlle Nortier, Pierre Duez and Caroline Stévigny

      Article first published online: 24 OCT 2013 | DOI: 10.1002/jat.2951

      Schizandrin and schizandrin B were tested on HK-2 cells for potential alleviation of cisplatin-mediated nephrotoxicity. Both compounds enhanced cell survival via reducing apoptosis rate. Schizandrin showed moderate effects towards modulation of regeneration capacities of healthy cells. Both Schi and Schi B limited extracellular matrix deposition and could help preventing dedifferentiation processes via the β-catenin pathway. Schizandrin and Schizandrin B present promising activities for future development of protective agents against CisPt nephrotoxicity.

    2. Borrelidin has limited anti-cancer effects in bcl-2 overexpressing breast cancer and leukemia cells and reveals toxicity in non-malignant breast epithelial cells

      Diana Gafiuc, Marlene Weiß, Ioannis Mylonas and Ansgar Brüning

      Article first published online: 24 OCT 2013 | DOI: 10.1002/jat.2946

      Borrelidin is a macrolide antibiotic with selective cytotoxicity against endothelial cells and leukemia cells. While testing the efficacy of borrelidin on breast cancer cells, we noticed the occurrence of non-specific toxicity in non-malignant cells and pre-existing resistance mechanisms against borrelidin in breast cancer and leukemia cell lines. This may limit the use of borrelidin as a possible new anti-cancer drug and raises concerns about possible unspecific toxicities.

    3. Prophylactic administration of non-organophosphate cholinesterase inhibitors before acute exposure to organophosphates: assessment using terbufos sulfone

      Dietrich E. Lorke, Syed M. Nurulain, Mohamed Y. Hasan, Kamil Kuča and Georg A. Petroianu

      Article first published online: 17 OCT 2013 | DOI: 10.1002/jat.2939

      In search of a broad-spectrum prophylactic agent for organophosphate poisoning, the reversible AChE-inhibitors pyridostigmine, physostigmine, ranitidine, tacrine and K-27 (an experimental oxime) were tested as a pre-treatment before exposure to the organophosphate AChE-inhibitor terbufos sulfone. Their efficacy in reducing organophosphorus compound-induced mortality was assessed by Cox survival analysis. Best in vivo protection from terbufos sulfone-induced mortality was achieved by K-27. Prophylactic administration of an oxime, such as K-27, in case of imminent organophosphorus compound exposure may be a viable option.

    4. Assessment of the genotoxic and carcinogenic potentials of 3-aminothiophene derivatives using in vitro and in silico methodologies

      Alban Lepailleur, Ronan Bureau, Marie-Pierre Halm-Lemeille, Michel Bouquet, Régis Pecquet, Christine Paris-Soubayrol, Jérémie Le. Goff, Véronique André, Yannick Lecluse, Pierre Lebailly, Marie-Aline Maire and Paule Vasseur

      Article first published online: 11 OCT 2013 | DOI: 10.1002/jat.2938

      The publication focus on the genotoxic/mutagenic and carcinogenic potentials of the methyl 3-amino-4-methylthiophene-2-carboxylate, a precursor of the articaine anesthetic. To discern some structure-toxicity relationships, we also studied the 3-amino 4-methylthiophene and the 2-acetyl 4-chlorothiophene. All these derivatives were negative in the Ames tests and the Comet assays. Only the 3-amino 4-methylthiophene showed a carcinogenic potential in the in vitro SHE CTA. In silico predictions of the reactivity of these derivatives towards CYP450 enabled us to hypothesize their possible toxicological pathways.

    5. Rapid determination of quetiapine in blood by gas chromatography–mass spectrometry. Application to post-mortem cases

      Olga López-Guarnido, María Jesús Tabernero, Antonio F. Hernández, Lourdes Rodrigo and Ana M. Bermejo

      Article first published online: 11 OCT 2013 | DOI: 10.1002/jat.2944

      A simple, fast and sensitive method for the determination of quetiapine (QTP) in human blood has been developed and validated. The method involved a basic liquid-liquid extraction procedure and subsequent analysis by gas chromatography-mass spectrometry (GC/MS), previous derivatization with bis(trimethylsilyl)-trifluoroacetamide (BSTFA) and chorotrimethylsilane (TMCS) (99:1). The method validation included linearity with a correlation coefficient >0.99 over the range 0.02–1 µg mL-1.The limit of detection was 0.005 µg mL-1. The procedure was further applied to post mortem cases.

    6. A catch-up validation study of an in vitro skin irritation test method using reconstructed human epidermis LabCyte EPI-MODEL24

      Hajime Kojima, Masakazu Katoh, Shinsuke Shinoda, Saori Hagiwara, Tamie Suzuki, Runa Izumi, Yoshihiro Yamaguchi, Maki Nakamura, Toshihiko Kasahawa and Aya Shibai

      Article first published online: 10 OCT 2013 | DOI: 10.1002/jat.2937

      According to the performance standard in Organisation for Economic Co-operation and Development (OECD) Test Guideline (TG) No. 439, the catch-up validation study for a skin irritation assay using reconstructed human epidermis (RhE) LabCyte EPI-MODEL24 (LabCyte EPI-MODEL24 SIT) were conducted at three laboratories in Japan. The obtained results showed that LabCyte EPI-MODEL24 SIT conformed more accurately to the classifications for skin irritation under the United Nations Globally Harmonised System of Classification and Labeling of Chemicals (UN GHS).

    7. Using quantitative structure–activity relationship modeling to quantitatively predict the developmental toxicity of halogenated azole compounds

      Evisabel A. Craig, Nina Ching Wang and Q. Jay Zhao

      Article first published online: 9 OCT 2013 | DOI: 10.1002/jat.2940

      Here we describe a QSAR model that may serve as an alternative approach to fill data gaps in the developmental toxicity profile of halogenated azole compounds. Chemical information from known developmental toxicants was curated using the OECD QSAR Toolbox and analyzed via linear regression to build a predictive model with an R2 of 0.79 and a Q2 of 0.77. Internal and external validation demonstrated the good predictability of the model, which estimated developmental LOAELs within 3 folds of experimental values.

    8. Toxic effects of a modified montmorillonite clay on the human intestinal cell line Caco-2

      Sara Maisanaba, Daniel Gutiérrez-Praena, Silvia Pichardo, F. Javier Moreno, María Jordá, Ana M. Cameán, Susana Aucejo and Ángeles Jos

      Article first published online: 9 OCT 2013 | DOI: 10.1002/jat.2945

      The increasing used of montmorillonite based clays in the development of reinforced food contact materials makes necessary to study the toxicological aspects of these materials. In the present study, the cytotoxic effects of a non-modified clay (Cloisite® Na+) and an organoclay (Cloisite® 30B) have been investigated in the intestinal cell line Caco-2. Only Cloisite® 30B showed cytotoxicity. In order to elucidate the toxic mechanisms underlying these effects, apoptosis, oxidative stress and genotoxicity biomarkers as well as ultrastructural alterations were studied.

    9. Histopathological findings on Carassius auratus hepatopancreas upon exposure to acrylamide: correlation with genotoxicity and metabolic alterations

      Miguel Larguinho, Pedro M. Costa, Gonçalo Sousa, Maria H. Costa, Mário S. Diniz and Pedro V. Baptista

      Article first published online: 7 OCT 2013 | DOI: 10.1002/jat.2936

      The toxicity of acrylamide to aquatic organisms is scarcely known. Goldfish were exposed to several concentrations of waterborne acrylamide and the results showed a dose-dependent increase in genotoxicity endpoints and in the levels of hepatic CYP1A and GST activity. In addition, acrylamide induced more histopathological changes to pancreatic acini than to the hepatic parenchyma. Overall, the findings suggest a specific mode of action that overran the defences against acrylamide a rather than causing a general systemic failure.

    10. Towards an integrated in vitro strategy for estrogenicity testing

      Si Wang, Jac M. M. J. G. Aarts, Laura H. J. de Haan, Dimitrios Argyriou, Ad A. C. M. Peijnenburg, Ivonne M. C. M. Rietjens and Toine F. H. Bovee

      Article first published online: 30 SEP 2013 | DOI: 10.1002/jat.2928

      In order to define an in vitro integrated testing strategy (ITS) for estrogenicity, a set of 23 reference compounds representing diverse chemical classes were tested in a series of in vitro assays including proliferation and reporter gene assays. All in vitro assays used revealed a reasonable to good correlation (R2 = 0.62–0.87) with the in vivo uterotrophic assay but the combination of the yeast estrogen bioassay with the U2OS ERα-CALUX assay seems most promising for an ITS for in vitro estrogenicity testing.

    11. Xenobiotic metabolism induction and bulky DNA adducts generated by particulate matter pollution in BEAS-2B cell line: geographical and seasonal influence

      Capucine Lepers, Véronique André, Mona Dergham, Sylvain Billet, Anthony Verdin, Guillaume Garçon, Dorothée Dewaele, Fabrice Cazier, François Sichel and Pirouz Shirali

      Article first published online: 30 SEP 2013 | DOI: 10.1002/jat.2931

      Particulate matter (PM) toxicity is of growing interest, but the link between chemical composition and toxicity remains unclear. We studied 6 native PM samples from various origins on BEAS-2B cell line. Cells exposure resulted in CYP1A1 and CYP1B1 genes induction, followed by an increase in EROD activity leading to bulky DNA-adducts. We showed that adducts level is only partly explained by PM content in polycyclic aromatic hydrocarbons, and that inorganic compounds may promote adducts formation by supporting CYPs activity.

    12. Circulating microRNAs as biomarkers of retinal toxicity

      Qinghai Peng, Wenhu Huang and Annette John-Baptiste

      Article first published online: 30 SEP 2013 | DOI: 10.1002/jat.2930

      Despite attrition due to retinal toxicity during drug development there are no early reliable predictive biomarkers of retinal toxicity. Currently the technologies for assessing retinopathy are limited to inconvenient visual behavioral tests, invasive electroretinograms or terminal histopathology. To address this gap, we explored several retinal enriched miRNAs in rats ex vivo and in vivo with pan-CDK inhibitors to assess these miRNAs in plasma. Although they need additional confirmatory evaluation, results provide promise for further testing using other retinal toxicants.

    13. Long-term maintenance of HepaRG cells in serum-free conditions and application in a repeated dose study

      Sebastian Klein, Daniel Mueller, Valery Schevchenko and Fozia Noor

      Article first published online: 30 SEP 2013 | DOI: 10.1002/jat.2929

      HepaRG cells were maintained viable with constant CYP activities and liver-specific functions for 30 days in serum-free medium (SFM). The long-term viability and CYP activities were superior to those reported for primary human hepatocytes in vitro. Metabolic flux analysis indicates that the central carbon metabolism in SFM is comparable to standard conditions. This system can be applied for repeated dose toxicity assessments and will pave way for application on high content ‘omics’ platform in systems toxicology.

    14. Identification of metabolomic biomarkers for drug-induced acute kidney injury in rats

      Takeki Uehara, Akira Horinouchi, Yuji Morikawa, Yutaka Tonomura, Keiichi Minami, Atsushi Ono, Jyoji Yamate, Hiroshi Yamada, Yasuo Ohno and Tetsuro Urushidani

      Article first published online: 30 SEP 2013 | DOI: 10.1002/jat.2933

      Metabolomics involves the comprehensive and simultaneous determination of endogenous or exogenous metabolites in biological samples. The present study aimed to identify metabolomic biomarkers that could provide early and sensitive indication of nephrotoxicity in rats. Metabolomic analyses were performed using CE-TOFMS on rat plasma treated with several nephrotoxicants. Among a total of 169 metabolites identified, 3-methylhistidine, 3-indoxyl sulfate, and guanidoacetate were selected as candidate biomarkers, which may prove to be valuable plasma biomarkers for monitoring nephrotoxicity in rats.

    15. RBE4 cells are highly resistant to paraquat-induced cytotoxicity: studies on uptake and efflux mechanisms

      V. Vilas-Boas, R. Silva, P. Guedes-de-Pinho, F. Carvalho, M. L. Bastos and F. Remião

      Article first published online: 18 SEP 2013 | DOI: 10.1002/jat.2926

      The effects of the toxic herbicide paraquat (PQ) to the blood–brain barrier (BBB) have been scarcely studied. It is shown that RBE4 cells (in vitro model of a rat's BBB) were highly resistant to PQ-induced cytotoxicity and that PQ used the choline-uptake system to enter RBE4 cells instead of the transporters of basic or neutral amino acids or the polyamines. Furthermore, PQ was extruded back to the extracellular medium by P-glycoprotein. However, these uptake/efflux studies only partially explain RBE4 resistance to PQ.

    16. Alteration in cellular viability, pro-inflammatory cytokines and nitric oxide production in nephrotoxicity generation by Amphotericin B: involvement of PKA pathway signaling

      F. D. França, A. F. Ferreira, R. C. Lara, J. V. Rossoni Jr, D. C. Costa, K. C. M. Moraes, C. A. Tagliati and M. M. Chaves

      Article first published online: 18 SEP 2013 | DOI: 10.1002/jat.2927

      The use of Amphotericin B is often limited as a result of adverse effects, especially nephrotoxicity. In this study we evaluate that the inhibition of the PKA pathway decreases cell death, changes in the production of pro-inflammatory cytokines and nitric oxide in LLC-PK1 and MDCK cell lines. Therefore, the inhibition of the PKA pathway can help decrease nephrotoxicity caused by Amphotericin B.

    17. Prenatal exposure to bisphenol A at the reference dose impairs mitochondria in the heart of neonatal rats

      Ying Jiang, Juan Liu, Yuanyuan Li, Huailong Chang, Gengqi Li, Bing Xu, Xi Chen, Weiyong Li, Wei Xia and Shunqing Xu

      Article first published online: 18 SEP 2013 | DOI: 10.1002/jat.2924

      Bisphenol A exposure is epidemiologically associated with heart disease. We found prenatal exposure to BPA at the reference dose induced mild ultrastructure changes in myocardial mitochondria at birth day. Mitochondrial membrane potential and the activity of respiratory chain complex II were significantly decreased. The expression levels of key regulators involved in mitochondria energy metabolism and ATP-generating pathways were down-regulated. The study demonstrated that prenatal exposure to BPA at the reference dose could impair mitochondria in the hearts of neonatal rats.

    18. Obese mice are resistant to eosinophilic airway inflammation induced by diesel exhaust particles

      Rie Yanagisawa, Eiko Koike, Takamichi Ichinose and Hirohisa Takano

      Article first published online: 18 SEP 2013 | DOI: 10.1002/jat.2925

      We investigated the differences in susceptibility to DEP-induced airway inflammation between obese mice (db/db) and lean mice (db/+m). DEP exposure results in eosinophilic airway inflammation via generation of Th2 cytokines, chemokines, and soluble ICAM-1 in db/+m mice; whereas in db/db mice, there was mild neutrophilic inflammation with attenuation of eosinophilic infiltration in the lungs. Obesity can affect the susceptibility to DEP-related airway inflammation, presumably because of differences in systemic and local inflammatory responses between obese and lean individuals.

    19. Differential responses to JNJ-37822681, a specific and fast dissociating dopamine D2 receptor antagonist, in cynomolgus monkey and Sprague–Dawley rat general toxicology studies: clinical observations, prolactin levels, mammary histopathology findings and toxicokinetics

      Eric J. de Waal, Maria Desmidt, Sven Korte, Marc Niehoff, Kevan Chase, Wayne Arrowsmith and Ann Lampo

      Article first published online: 16 SEP 2013 | DOI: 10.1002/jat.2916

      JNJ-378226 81 is a potent, specific and fast dissociating dopamine D2 receptor antagonist intended for the treatment of schizophrenia. Differences between Cynomolgus monkeys and Sprague-Dawley rats were noted in their response to JNJ-37822681 in terms of extrapyramidal (EPS)-like clinical signs and prolactin-mediated tissue changes in the mammary gland. Overall the Cynomolgus monkey showed better predictivity towards the nature of JNJ-37822681-associated adverse events in schizophrenia patients than the Sprague-Dawley rat.

    20. CKD-501, a novel selective PPARγ agonist, shows no carcinogenic potential in ICR mice following oral administration for 104 weeks

      Kyoung-Sik Moon, Ji-Eun Lee, Hee Su Lee, In-Chang Hwang, Dal-Hyun Kim, Hyun-Kyu Park, Hyun-Ji Choi, Woori Jo, Woo-Chan Son and Hyo-In Yun

      Article first published online: 11 SEP 2013 | DOI: 10.1002/jat.2918

      CKD-501 is a peroxisome proliferator-activated receptor gamma (PPARγ) agonist that is effective for diabetes. Carcinogenicity study was conducted over a period of 104 weeks in ICR mice with three groups, each consisting of 60 mice in both sexes, received oral dosages of 0.2, 1.0 or 6.0 mg/kg/day. No significant treatment-related effects were observed on the survival rates. Expected non-neoplastic findings for this class of compound (adipocyte proliferation, bone marrow hypoplasia cardiomyopathy) were seen. No treatment-related neoplastic changes were observed.

  12. Short Communications

    1. LTMap: a web server for assessing the potential liver toxicity by genome-wide transcriptional expression data

      Li Xing, Leihong Wu, Yufeng Liu, Ni Ai, Xiaoyan Lu and Xiaohui Fan

      Article first published online: 11 SEP 2013 | DOI: 10.1002/jat.2923

      LTMap is a web server for assessing the potential liver toxicity of compounds by comparing gene expression profile.

  13. Research Articles

    1. Effects of butyltins on mitogen-activated-protein kinase kinase kinase and Ras activity in human natural killer cells

      Lindsay J. Celada and Margaret M. Whalen

      Article first published online: 5 SEP 2013 | DOI: 10.1002/jat.2921

      Butyltins (BTs) contaminate the environment and are found in human blood. Natural killer (NK) cells defend against tumors and virally-infected cells. Mitogen-activated protein kinases (MAPKs) regulate NK cells. BTs activate MAPKs in human NK cells. The current study examined if BT-induced activation of MAPK kinase kinases (MAP3Ks) caused MAPK activation. Tributyltin (TBT) activated the MAP3K, c-Raf, within 10 minutes and the MAP3K, ASK1, after one hour. Results suggest that both TBT and dibutyltin (DBT) are impacting the regulation of c-Raf.

    2. Comparison of inhibitory effects between acetaminophen–glutathione conjugate and reduced glutathione in human glutathione reductase

      Erika Nýdlová, Martina Vrbová, Petr Česla, Barbora Jankovičová, Karel Ventura and Tomáš Roušar

      Article first published online: 5 SEP 2013 | DOI: 10.1002/jat.2914

      Acetaminophen overdose is the most frequent cause of acute liver injury. The oxidation product reacts with glutathione generating acetaminophen-glutathione conjugate (APAP-SG). The aim of our study was to estimate the toxicity of APAP-SG by characterizing the inhibitory effect in glutathione reductase (GR) and comparing that to the inhibitory effect of reduced glutathione. Our results show that GR was significantly inhibited in the presence of both substances. The type of enzyme inhibition was observed to be competitive in the both cases.

    3. Proteomic profiling in incubation medium of mouse, rat and human precision-cut liver slices for biomarker detection regarding acute drug-induced liver injury

      Rachel P. L. van Swelm, Mackenzie Hadi, Coby M. M. Laarakkers, Rosalinde Masereeuw, Geny M. M. Groothuis and Frans G. M. Russel

      Article first published online: 30 AUG 2013 | DOI: 10.1002/jat.2917

      Drug-induced liver injury often leads to drug withdrawal. Proteins released from human, mouse and rat precision-cut liver slices (PCLS) exposed to several hepatotoxicants were profiled using MALDI-TOF MS. Following acetaminophen treatment, similar changes in protein profiles (including the key proteins SOD1, CA3 and calmodulin) were demonstrated as previously found in vivo in mouse urine. Incubation with diclofenac resulted in the same toxicity parameters as observed for acetaminophen. In conclusion, proteomics can be applied to identify potential translational biomarkers in PCLS.

    4. In vitro protection by pyruvate against cadmium-induced cytotoxicity in hippocampal HT-22 cells

      Ethan Poteet, Ali Winters, Luokun Xie, Myoung-Gwi Ryou, Ran Liu and Shao-Hua Yang

      Article first published online: 30 AUG 2013 | DOI: 10.1002/jat.2913

      Cadmium is a toxic metal that inhibits glycolysis indirectly through its actions on the mitochondria. Pyruvate restores glycolysis and mitigates the inhibitory action of cadmium in mitochondria and glycolysis. Our study provided the first in vitro evidence that pyruvate protects against the cytotoxic action of cadmium and that pyruvate might be a potential therapy for cadmium poisoning.

    5. The alkylphenols 4-nonylphenol, 4-tert-octylphenol and 4-tert-butylphenol aggravate atopic dermatitis-like skin lesions in NC/Nga mice

      Kaori Sadakane, Takamichi Ichinose, Hirohisa Takano, Rie Yanagisawa, Eiko Koike and Ken-ichiro Inoue

      Article first published online: 23 AUG 2013 | DOI: 10.1002/jat.2911

      Alkylphenols (4-nonylphenol, 4-tert-octylphenol, and 4-tert-butylphenol) exacerbated mite allergen-induced atopic dermatitis in a mouse model. Alkylphenols tended to increase immunoglobulin levels in serum and increased the expression of inflammatory cytokines in lesions. In contrast, the levels of T-helper 1 cytokines decreased. The uptake of very low levels of alkylphenols may contribute to the current increase in atopic dermatitis.

    6. Effects of water soluble PM2.5 extracts exposure on human lung epithelial cells (A549): A proteomic study

      Qingyu Huang, Jie Zhang, Siyuan Peng, Meiping Tian, Jinsheng Chen and Heqing Shen

      Article first published online: 14 AUG 2013 | DOI: 10.1002/jat.2910

      Airborne particulate matter (PM)2.5 exposure has been associated with a number of adverse health effects. However, the mechanisms underlying PM2.5 toxicity remain unclear. In the current proteomic study, 22 differentially expressed proteins responsive to water-soluble PM2.5 extract treatment in A549 cells were identified by 2-D difference gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry. Furthermore, induction of apoptosis was proposed as a vital mechanism for water-soluble PM2.5 extract cytotoxicity. These results may provide useful insights into the mechanisms of PM2.5 toxicity and assist in the development of biomarkers of PM2.5 exposure.

    7. DNA fragmentation induced by all-trans retinoic acid and its steroidal analogue EA-4 in C2C12 mouse and HL-60 human leukemic cells in vitro

      Raghda S. Alakhras, Georgia Stephanou, Nikos A. Demopoulos, Konstantinos Grintzalis, Christos D. Georgiou and Sotirios S. Nikolaropoulos

      Article first published online: 2 AUG 2013 | DOI: 10.1002/jat.2908

      We (a) investigated the ability of all-trans retinoic acid and its steroidal analogue EA-4 to induce DNA fragmentation by using single cell gel electrophoresis, and (b) quantified the ability of the retinoids under study to induce necrotic/apoptotic small size (0–1 kb) fragmented DNA. We found that they (a) provoke DNA migration due to DNA fragmentation, and (b) induce significantly small fragments of genomic DNA in C2C12 mouse and HL-60 human leukemic cells, as indicated by the quantification of necrotic/apoptotic small DNA fragments.

    8. Characterization of the components of urban particulate matter mediating impairment of nitric oxide-dependent relaxation in intrapulmonary arteries

      Arnaud Courtois, Caroline Prouillac, Isabelle Baudrimont, Céline Ohayon-Courtes, Véronique Freund-Michel, Mathilde Dubois, Mariette Lisbonne-Autissier, Roger Marthan, Jean-Pierre Savineau and Bernard Muller

      Article first published online: 23 JUL 2013 | DOI: 10.1002/jat.2909

      We have previously shown that exposure to urban particulate matter (UPM) impairs endothelial nitric oxide (NO) bioactivity in intrapulmonary arteries. Since UPM is composed of heterogeneous constituents, the present study was designed to clarify the class of pollutants being responsible for such effect. We showed that UPM-induced impairment of endothelial NO bioactivity unlikely involved a soluble factor released by vascular cells, but organic extractible and acidic-sensitive constituents of UPM. Among those constituents, endotoxin may be responsible for such impairment.

    9. Induction of apoptosis by antimycin A in differentiated PC12 cell line

      Lanju Xu, Jing Xu, Shichang Liu and Zhuo Yang

      Article first published online: 19 JUL 2013 | DOI: 10.1002/jat.2890

      The aim of the present study was to investigate the effect of AMA on PC12 cells in vitro. Results of the nuclear morphology and the flow cytometer indicated that AMA efficiently induced PC12 cell apoptosis. Moreover, the levels of ROS and Ca2+ increased by AMA treatment. It was found that the Ca2+ chelators and ROS scavengers could delay AMA-induced PC12 cell apoptosis. In conclusion, the present study found that AMA induced PC12 cell apoptosis through ROS and Ca2+.

    10. Phthalates efficiently bind to human peroxisome proliferator activated receptor and retinoid X receptor α, β, γ subtypes: an in silico approach

      M. K. Sarath Josh, S. Pradeep, K. S. Vijayalekshmi Amma, S. Balachandran, U. C. Abdul Jaleel, Mukesh Doble, Friedrich Spener and Sailas Benjamin

      Article first published online: 11 JUL 2013 | DOI: 10.1002/jat.2902

      This exhaustive in silico study looks into the molecular interactions of 15 hazardous diphthalates, their metabolites vis-á-vis certain natural and synthetic ligands with nuclear hPPAR and hRXR receptor subtypes via molecular docking using Glide. DIDP showed highest G score with hPPARγ, while DEHP generally showed a greater preference to hRXRs than hPPARs and showed highest G score with hRXRα. These in silico evidences would lead to further in vitro mechanistic as well as in vivo preclinical and clinical studies.

    11. Role of methotrexate exposure in apoptosis and proliferation during early neurulation

      Xiuwei Wang, Jianhua Wang, Tao Guan, Qian Xiang, Mingsheng Wang, Zhen Guan, Guannan Li, Zhiqiang Zhu, Qiu Xie, Ting Zhang and Bo Niu

      Article first published online: 9 JUL 2013 | DOI: 10.1002/jat.2901

      In this study, we established an animal model of neural tube defects (NTDs) by use of MTX to investigate the role of apoptosis and proliferation in NTDs. We concluded that MTX caused a folate and folate-associated dysmetabolism, and further induced abnormal apoptosis and proliferation, which may play a critical role in the occurrence of NTDs caused by folate deficiency.

    12. Cytotoxic effects of the quinolone levofloxacin on rabbit meniscus cells

      Linlong Wang, Yunpeng Wu, Yang Tan, Xi Fei, Yu Deng, Hong Cao, Biao Chen, Hui Wang, Jacques Magdalou and Liaobin Chen

      Article first published online: 1 JUL 2013 | DOI: 10.1002/jat.2903

      Rabbit meniscus cells were administrated with different concentrations of levofloxacin. After treatment with levofloxacin for 48 h, cells viability was decreased and apoptotic cells increased. Meanwhile, active caspase-3 protein expression level was also increased and mRNA expression level of Bcl-2 decreased. Both mRNA and protein expression levels of MMP-3 and MMP-13 increased. Besides, the mRNA expression levels of TIMP-3 and col1a1 were decreased, while that of iNOS increased. Thus, levofloxacin induced apoptosis and unbalanced MMPs/TIMPs expression in meniscus cells.

    13. Combination effects of amphetamines under hyperthermia - the role played by oxidative stress

      Diana Dias da Silva, Elisabete Silva and Helena Carmo

      Article first published online: 13 JUN 2013 | DOI: 10.1002/jat.2889

      The impact of hyperthermia on the cytotoxicity of combinations of MDMA and three other amphetamines often co-ingested, d-amphetamine, methamphetamine and 4-methylthioamphetamine, was evaluated. In hyperthermia (40.5 °C), amphetamine mixtures produced deleterious effects, even when individual drugs were combined at negligible concentrations. Mixture effects were accurately predicted by concentration addition model. RNS/ROS production, GSH and ATP depletion and mitochondrial impairment were strongly exacerbated under hyperthermia, emphasizing the potential increased risks of ecstasy intake, especially when hyperthermia occurs concurrently with polydrug abuse.

    14. N-hexane inhalation during pregnancy alters DNA promoter methylation in the ovarian granulosa cells of rat offspring

      Hong Li, Jin Liu, Yan Sun, Wenxiang Wang, Shaozheng Weng, Shihua Xiao, Huiling Huang and Wenchang Zhang

      Article first published online: 6 JUN 2013 | DOI: 10.1002/jat.2893

      This paper carried out the MeDIP-Chip method to investigate the global genomic promoter methylation status of F1 Wistar rats' Ovarian granulosa cells after maternal n-hexane exposure. The Gene Ontology Consortium, the DAVID Functional Annotation Clustering Tool, hierarchical clustering, and KEGG pathway were used to delineate the altered methylation of promoters of gene and find out the gene-enriched clusters.

    15. In vitro PFOS exposure on immune endpoints in bottlenose dolphins (Tursiops truncatus) and mice

      Jena R. Wirth, Margie M. Peden-Adams, Natasha D. White, Gregory D. Bossart and Patricia A. Fair

      Article first published online: 30 MAY 2013 | DOI: 10.1002/jat.2891

      Following in vitro exposure to environmentally relevant perfluorooctane sulfonate (PFOS) concentrations (0–5 µg ml−1) in dolphin peripheral blood leukocytes (PBLs) and adult female B6C3F1 mouse splenocytes, lymphocyte proliferation and natural cell killer activity was measured. Risk assessment using a parallelogram approach indicated that dolphin PBLs exhibited similar results to correlative field studies; mice were generally more sensitive than dolphins; dolphins were more similar to humans than mice. The parallelogram approach was predictive two-thirds of the time for dolphin response.

    16. Dermal irritation of petrolatum in rabbits but not in mice, rats or minipigs

      S. A. Chandra, R. A. Peterson, D. Melich, C. M. Merrill, D. Bailey, K. Mellon-Kusibab and R. Adler

      Article first published online: 30 MAY 2013 | DOI: 10.1002/jat.2895

      New Zealand white rabbits treated with white petrolatum for 2 weeks had mild acanthosis, hyperkeratosis, dermal edema with mixed inflammatory cells consistent with dermal irritation. Wistar-Han rats, CD1 mice, C57/BL/6J mice, and Gottingen minipigs did not exhibit clinical or histologic evidence of dermal irritation to petrolatum.

    17. Inhibition of cytochrome P450s enhances (+)-usnic acid cytotoxicity in primary cultured rat hepatocytes

      Qiang Shi, James Greenhaw and William F. Salminen

      Article first published online: 20 MAY 2013 | DOI: 10.1002/jat.2892

      (+)-Usnic acid (UA) may cause hepatotoxicity. We examined if its metabolism plays a role in toxicity in primary rat hepatocytes. When cytochrome P450 (CYP) activity was suppressed by pan-CYP inhibitor SKF-525A, or CYP1A inhibitor alpha-naphthoflavone, or CYP3A inhibitor ketoconazole, UA induced inhibition of cellular respiration, adenosine triphosphate depletion and cytotoxicity was significantly increased. The CYP2B/2C inhibitor ticlopidine showed no effects. We suggest UA is bio-transformed to less toxic metabolites in rat hepatocytes, mainly by CYP1A and 3A, but not 2B/2C.

    18. Nonylphenol-induced apoptotic cell death in mouse TM4 Sertoli cells via the generation of reactive oxygen species and activation of the ERK signaling pathway

      Mi-Sun Choi, Han-Jin Park, Jung-Hwa Oh, Eun-Hee Lee, Se-Myo Park and Seokjoo Yoon

      Article first published online: 16 MAY 2013 | DOI: 10.1002/jat.2886

      Nonylphenol (NP), a representative endocrine disruptor, interferes with reproductive function in aquatic organisms and animals. Although many studies have focused on apoptotic cell death by NP, the fundamental mechanism of NP on apoptosis remains poorly understood. Here, we investigated the molecular mechanism on NP-induced apoptotic cell death in mouse TM4 Sertoli cells. Our results suggest that NP induced apoptotic cell death is mediated via ROS generation and activation of the ERK signaling pathway in TM4 cells.

    19. Mixtures of 3,4-methylenedioxymethamphetamine (ecstasy) and its major human metabolites act additively to induce significant toxicity to liver cells when combined at low, non-cytotoxic concentrations

      Diana Dias da Silva, Elisabete Silva, Félix Carvalho and Helena Carmo

      Article first published online: 14 MAY 2013 | DOI: 10.1002/jat.2885

      Combination effects between methylenedioxymethamphetamine (MDMA; ecstasy) and its metabolites methylenedioxyamphetamine, α-methyldopamine and N-methyl-α-methyldopamine were evaluated in Hep G2 cells at normal (37 ºC) and hyperthermic (40.5  ºC) incubation conditions. Joint cytotoxic effects measured as viability in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay were predicted using the concentration addition and independent action models. The combination of MDMA and its metabolites induced additive toxicity that was accurately predicted by the concentration addition model. Substantial combination effects were noted for mixtures at individually non-cytotoxic concentrations. Hyperthermic incubations dramatically increased individual and joint cytotoxicity.


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