Journal of Applied Toxicology

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.

We investigated the effects of single-strand DNA-multi-walled carbon nanotubes (ss-DNA-MWCNT) on the rat spleen, after intraperitoneal (i.p.) administration. Carbon nanotubes were detected in the spleen using Raman spectroscopy, histopathologic examination, confocal microscopy and transmission electron microscopy (TEM), at different time points after administration. The dynamics of oxidative and nitrosative stress parameters, pro-inflammatory cytokines and the number of cells expressing caspase 3 and proliferating cell nuclear antigen were assessed. Our results indicate that MWCNT translocate progressively in the spleen, determine lymphoid hyperplasia and transient alterations of oxidative stress and inflammation.

The current challenge in safety assessments is to find ways to integrate mechanistic insight, in vitro data, in silico and in chemico information to predict in vivo effects. This study follows up on our proof of concept work and presents an updated Integrated Testing Strategy (ITS) to assess skin sensitization potency. The refined ITS predicted correctly 95% and 86% of chemicals in a test set (n = 21) for hazard and local lymph node assay (LLNA) potency classes, respectively. The study includes a detailed validation analysis and illustrations of practical applications.

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.

It is truly important to develop computational models to screen out hepatotoxic compounds in drug development. We have developed rigorously validated hybrid models using chemical descriptors and in vitro cell-imaging information which can be used in virtual screening of lead compounds with potential hepatotoxicity. Our results showed chemical structure and in vitro biological data can be complementary in enhancing the prediction accuracy of human hepatotoxicity and can afford rational mechanistic interpretation.

Studies on the effects of nanomaterial exposure in mammals are limited, and new methods for rapid risk assessment of nanomaterials are urgently required. The utility of C. elegans as an alternative model for toxicity screens was evaluated using 10 nanometer silver (10nmAg). Consistent with studies using mammalian in vitro and in vivo models, 10nmAg at 10 μg ml-1 reduced growth and induced oxidative damage to DNA, and ionic silver was found to be more toxic than 10nmAg to C. elegans.

Six different inbred rat strains were used to determine whether immune activation and progress of fibrosis after lung exposure of the alkylating agent melphalan are genetically regulated. Results demonstrate that timing and magnitude of airway inflammation and late collagen production in lungs are genetically influenced. Two rat strains which developed marked fibrosis also contained increased levels of cytotoxic T cells and natural killer (NK) cells in airways, suggesting that susceptibility to develop lung fibrosis is associated with high cytotoxic cell responses.

In light of the adverse reports of Bisphenol A (BPA) on reproduction, it was of interest to decipher the effect of neonatal exposure to BPA on Steroid receptors (SRs) and their coregulators in male reproduction. Developmental expressions in testis were analyzed by qPCR and Immunohistochemistry. SRs and coactivators perturbations at transcript and the protein level were observed, whereas corepressor expression remained unaltered in the BPA-exposed groups. These impairments can be a putative mechanism of adversities on fertility caused by BPA.

The aim of this study was to highlight the risks of long term exposure to low cadmium concentrations in hares (Lepus europaeus Pallas) as model organisms in a field study. The median cadmium concentrations (wet weight) in the muscle, liver, kidney, and brain of hares from exposed group ranged from 0.033-0.037, 0.763-1.054, 3.090-16.594, and 0.016-0.087 µg/g respectively. Expression of the metallothioneins I+II proteins was observed in tissues. Lipid peroxidation levels, measured as malondialdehyde (MDA) equivalents, increased with the cadmium concentration.

The aim of this study was to investigate the effects and mechanisms of syringin on lipopolysaccharide (LPS) and D-galactosamine (D-GalN)-induced fulminant hepatic failure (FHF) in mice. Pretreatment mice with syringin alleviated LPS/D-GalN-induced mortality and liver injury, inhibited hepatocellular apoptosis, reduced hepatic tissues tumor necrosis factor-a (TNF-α) production, and NF-kB activation. Our results indicated that syringin might alleviate the FHF induced by LPS/D-GalN through inhibiting NF-kB activation to reduce TNF-α production.

To investigate renal sensitivity to uranium exposure during development, we examined uranium distribution and uranium-induced apoptosis in the kidneys of neonate (7-day-old), prepubertal (25-day-old), and adult (70-day-old) male Wistar rats. Our findings suggest that age-at-exposure and exposure level are important parameters for uranium toxicity; uranium tends to persist in developing kidneys after low-level exposures, although renal toxicity is more pronounced in adults.

When added on murine MC3T3-E1 preosteoblasts, single-walled carbon nanotubes (SWCNTs) produced by radio frequency induction thermal plasma with three catalyst mixtures drastically reduced cell viability in a dose-dependent manner. In contrast, the viability of preosteoblasts seeded on SWCNTs was slightly decreased at 24 h without apoptosis and the cells could proliferate within 48 h. Thus except mechanical disturbance, thermal plasma grown SWCNTs induced no severe cytotoxicity on preosteoblasts and therefore could be more deeply studied for bone applications.

Cytotoxic, genotoxic and the hemolytic effects of titanium dioxide (TiO2) nanoparticles (~100 nm) was studied in human erythrocyte and lymphocyte cells, in vitro. TiO2 nanoparticle resulted in membrane damage, reduction of mitochondrial function. The particles induced DNA damage leading to apoptosis. Hemolytic property of TiO2 nanoparticle was characterized by spherocytosis and echinocytosis. Spectral analysis revealed a hemoglobin-TiO2 nanoparticle interaction.

In view of their potential application in oral drug delivery, we investigated the effect of halloysite nanotubes in an in vitro model of the large intestine. Toxicity tests and label-free quantitative mass spectrometric analysis indicate that halloysite exhibits a high degree of biocompatibility characterized by an absence of cytotoxicity, despite elevated proinflammatory cytokine release. Bioinformatic analysis of differentially expressed protein profiles suggest that halloysite stimulates processes related to cell growth and proliferation, subtle responses to cell infection, irritation and injury, enhanced antioxidant capability, and an overall adaptive response to exposure.

Alternative tests for skin sensitization under prevalidation address three endpoints: surface marker expression on dendritic cell lines, peptide reactivity and activation of Nrf2. Here we report a database on 145 chemicals measured in a dendritic cell activation test, the direct peptide reactivity assays and the KeratinoSens^TM assay. This comprehensive database may serve to develop a data-driven integrated testing strategy and serve as reference database when benchmarking new molecules with in vitro based read-across.

We investigated the effects of Asian sand dust (ASD) particles on respiratory and immune system in vitro. ASD enhanced the production of interleukin (IL)-6, IL-8 and ICAM-1 from human airway epithelial cells, as well as the expression of DEC205 on bone marrow-derived dendritic cells (BMDC) and the proliferation of splenocytes. Moreover, these effects differed partly by ASD events or by components adhered to ASD. These results suggest that ASD can contribute to the respiratory and immune health.

Cardiotoxicity is an important issue in drug development. In the present study, we focused on 8 genes (Spp1, Fhl1, Timp1, Serpine1, Bcat1, Lmcd1, Rnd1 and Tgfb2) as candidate genomic biomarkers for cardiotoxicity in rats, and constructed an optimized support vector machine (SVM) model that was composed of Spp1 and Timp1. This multi-gene model exhibited advantages over classic tools commonly used for cardiotoxicity evaluations in rats, and the application of the model could potentially lead to the production of safer drugs.

In the present study, we perform the first systematic reference gene screening for normalization of plasma miRNA quantitative real-time PCR (RT-qPCR) data in a rat model of acetaminophen-induced hepatotoxicity. By employing four independent algorithms, we evaluated the expression stability of six candidate reference genes and results show that U6 and 5S, two of the most commonly used housekeeping genes, were not ideal reference genes and miR-103 was verified to be a most stable normalizer for circulating microRNAs analysis in rats with acetaminophen-induced liver injury.

This study comparatively evaluated the influence of surface coating [citrate(Cit) vs 11-mercaptoundecanoic acid(11-MUA)] on the uptake and toxicity of gold nanoparticles(AuNPs) in HepG2 cells. Both Cit- and MUA-AuNPs were observed either inside intracellular endosomes or in intercellular spaces. Under the tested conditions, they did not induce cytotoxicity. However, an inverse concentration-dependent increase of DNA damage was observed in Cit-AuNPs- but not in MUA-AuNPs-exposed cells. These data demonstrate the impact of surface properties in the biocompatibility and safety of AuNPs.

Effects of AFB₁ on splenic lymphocyte phenotypes and cytokine expression were investigated in male F344 rats. Single-dose exposure to AFB₁ decreased expression of CD8⁺ T cells and CD3^-CD8a⁺ NK cells. Expressions of IL-4 and IFN-γ by CD4⁺ T cells, IL-4 and IFN-γ by CD8a⁺ cells, and TNF-α by NK cells were also inhibited; Repeated AFB₁ exposure significantly decreased expression of IL-4 by CD4⁺ T cells and significantly increased expressions of pro-IFN-γ by CD4⁺ T cells and TNF-α by NK cells.

This study investigated the role of oxidative stress in enantiomer-specific, profenofos (PFF)-induced cytotoxicity and genotoxicity in PC12 cells. A concentration- and time-dependent significant reduction of cell viability and induction of DNA damage, reactive oxygen species, malondialdehyde and gene expression was observed in (−)PFF, while (+)PFF and rac-PFF exhibited these effects to lesser degrees. Pre-treatment with vitamin E (600 μM) caused a significant attenuation in the toxic effect, further strengthening the involvement of oxidative stress in PFF mediated toxicity.

A vast majority of toxic agents target internal organs for their toxicity. The difficulty in obtaining human internal target organs to determine toxicity is a major obstacle in preventing diseases potentially resulting from exposure to toxic agents. Peripheral blood gene expression profiling has been hypothesized as a practical, highly sensitive and mechanistically relevant surrogate approach to determine target organ toxicity. Recent developments in this emerging toxicology research area, with a special emphasis on hepatotoxicity and pulmonary toxicity, are discussed in this review article.

Zinc oxide (ZnO) nanoparticles (NPs) have been used for diverse applications including paints, cosmetics, biomedicine, and food. ZnO NPs can cause pancreatitis and anemia and this might be an important data for toxicity and risk assessment of ZnO NP in subchronic 13-week toxicity study vial oral route in rats. The bio-persistence of ZnO NPs after ingestion is key to their toxicity; the no-observed-adverse effect level (NOAEL) of ZnO NPs was found to be 268.4mgkg^–1 per day for both sexes.

This study was designed to evaluate the genotoxicity of citalopram at the recommended human doses in somatic cells of mice. Mice exposed to citalopram at varying oral doses of 12 or 24 mg/kg for 7 days, exhibited significant increase in the level of DNA-strand breaking and micronuclei formation. Furthermore, by FISH analysis with the centromeric DNA-probe for erythrocyte micronuclei it could be shown that citalopram is aneugen as well as clastogen in somatic cells in vivo.

Amorimiaseptentrionalis contains sodium monofluoroactetate and can cause acute heart failure in ruminants when ingested in toxic doses. In this study we demonstrate that resistance to poisoning by A. septentrionalis can be improved in goats by the repeated administration of non-toxic doses of A. septentrionalis. We also show that increased resistance to poisoning by A. septentrionalis can also be achieved by the transfaunation of ruminal content from goats previously conditioned to beresistant to naïve goats.

EtHg toxicity differs from that of meHg, leading to different toxicity risks. In vitro studies comparing etHg with meHg demonstrate equivalent measured outcomes for cardiovascular, neural, and immune cells. However, distinct toxicokinetic profile between meHg and etHg, results in different compartmental distribution and shorter blood half-life for etHg which can be explained by a faster in vivo dealkylation of etHg than meHg. Immunotoxicity is more pronounced and more common for thimerosal etHg.

Advances in pediatric and obstetric surgery have resulted in an increase in the complexity, duration, and number of anesthetic procedures. It has been reported that prolonged exposure of the developing brain to clinically relevant concentration of anesthetics that have NMDA antagonist or GABA-mimetic properties, and/or their combinations, resulted in an extensive abnormal pattern of neuroapoptosis, and subsequent cognitive deficits in animals. Molecular imaging using positron emission tomography (PET) is a leading modality for obtaining non- or minimally invasive in vivo measurements of multiple biological processes in various organs. The main focus of this review is to describe molecular imaging approaches that have been used in the study of pediatric anesthetic-induced neuronal toxicity.

Metals can cause an increase in reactive oxygen species (ROS) accumulation resulting in programmed cell death. The exact role that zinc (Zn) plays in the regulation of apoptosis remains ambiguous. Intracellular free Zn modulates p53 activity and stability, excess Zn alters p53 protein structure and down-regulates binding to DNA. Copper (Cu) accumulation causes apoptosis that is mediated by DNA damage and subsequent p53 activation. This review discusses a central role of Zn and of p53 in cellular stress responses induced by redox active biometal Cu.

In the present study, we examined the role that chronic ethanol (EtOH) exposure plays in gene expression changes during early stage of embryonic differentiation. By employing a transcriptomic approach, we have identified genes, including GSC, which are vulnerable to EtOH exposure during the early stages of embryonic development. We reported that EtOH deregulates GSC expression by altering nodal signaling pathway, which may provide a new avenue for analyzing the molecular mechanisms behind EtOH teratogenicity in FASD individuals.

A new reactive oxygen species (ROS) assay using serum albumin (aROS assay) was developed for photosafety assessment in biomimetic solution, and validation study demonstrated the high robustness of the aROS assay and improved applicability to poorly soluble chemicals. Assessments on 31 test samples were also indicative of good predictive capacity of the aROS assay. The aROS assay may allow photosafety assessment of new drug entities with a wide range of applicability partly under experimental biomimetic conditions.

Drinking water contaminated with 100 ppm CdCl₂ promoted non-rapid-eye-movement (REM) sleep and reduced locomotor activities during the active phase of rats. The duration of CdCl₂ exposure required to modify these behaviors (28 h) was far shorter than that to produce apparent hepatic/renal damage by cadmium ingestion. Because CdCl₂ administration increased the cerebral concentration of oxidized glutathione, we suggest cadmium-induced sleep as a response for oxidative stress clearance. We propose that facilitated non-REM sleep could be a behavioral marker for CdCl₂ poisoning.

In this study we assessed cardiovascular toxicity of seven known human cardiotoxic drugs (aspirin, clomipramine hydrochloride, cyclophosphamide, nimodipine, quinidine, terfenadine and verapamil hydrochloride) and two non-cardiovascular toxicity drugs (gentamicin sulphate and tetracycline hydrochloride) in zebrafish larvae using six specific phenotypic endpoints: heart rate, heart rhythm, pericardial edema, circulation, hemorrhage and thrombosis. All the tested drugs were delivered into zebrafish by direct soaking and yolk sac microinjection, respectively, and cardiovascular toxicity was quantitatively or qualitatively assessed at 4 and 24 h post drug treatment. The results showed that aspirin accelerated the zebrafish heart rate (tachycardia), whereas clomipramine hydrochloride, cyclophosphamide, nimodipine, quinidine, terfenadine and verapamil hydrochloride induced bradycardia. Quinidine and terfenadine also caused atrioventricular (AV) block. Nimodipine treatment resulted in atrial arrest with much slower but regular ventricular heart beating. All the tested human cardiotoxic drugs also induced pericardial edema and circulatory disturbance in zebrafish. Therewas no sign of cardiovascular toxicity in zebrafish treated with non-cardiotoxic drugs gentamicin sulphate and tetracycline hydrochloride. The overall prediction success rate for cardiotoxic drugs and non-cardiotoxic drugs in zebrafish were 100% (9/9) as compared with human results, suggesting that zebrafish is an excellent animal model for rapid in vivo cardiovascular toxicity screening.

ZnO nanoparticles are mass-produced nanomaterials. In this study, we evaluated the acute toxic effects of 30 nm-diameter ZnO nanoparticles after intranasal instillation by behavioral tests, magnetic resonance imaging and histology. It was shown ZnO nanoparticles instilled into the nasal cavity caused significant structural damages to rat olfactory epithelium (OE) and olfactory impairment. ZnO NP-induced OE damages are characterized by edema, degeneration of epithelial cells and increased blood vessel permeability. The possible toxicological mechanism might involve cellular energy metabolic dysfunction.

Renal complications are often detected in patients with inflammatory bowel disease (IBD). Because renal conventional markers are not sensitive and/or specific, a new renal biomarker is needed. We have recently identified urinary vanin-1 as an early biomarker for the detection of nephrotoxicant-induced renal injury. In this study, we compared the usefulness of urinary vanin-1 with other newly developed biomarkers for the detection of renal complications in rats with experimental colitis.

We evaluated the effectiveness of a novel synthetic compound, 3-Amino-3-(4-fluoro-phenyl)-1H-quinoline-2,4-dione (KR-22335) as an otoprotective agent against cisplatin. The effect of KR-22335 was tested in HEI-OC1 cells and in a zebrafish model. KR-22335 inhibited cisplatin-induced apoptosis, mitochondrial injury, and activation of JNK, p-38, caspase-3 and PARP in HEI-OC1 cells. KR-22335 prevented cisplatin-induced destruction of zebrafish neuromasts. The results of this study suggest that KR-22335 may prevent ototoxicity caused by cisplatin through the inhibition of mitochondrial dysfunction and suppression of ROS generation.

Previously, we identified the novel binding motif of aryl hydrocarbon receptor (AhR), “AhR-responsive element III (AHRE-III)” upstream from the tyrosine hydroxylase (TH) gene. We investigated whether or not 2,3,7,8”tetrachlorodibenzo-p-dioxin (TCDD) could regulate AHRE-III transcriptional activitywithin TH-immunoreactive (ir) neurons of midbrain dopaminergic nuclei. Using transgenic mice, it was demonstrated that fetal exposure to TCDD significantly increased TH-ir intensity and the numbers of TH-ir neurons within midbrain at 8 wk. It is suggested that this increase is caused via AhR-AHRE-III-mediated pathway.

The adverse effects of methyllycaconitine (MLA) have been attributed to its competitive antagonism of nicotinic acetylcholine receptors (nAChRs). The objective of this study was to characterize the role of the α₇ subunit of nAChRs in the acute toxicosis of MLA by comparing the lethal dose (LD₅₀) of MLA in wild-type mice to mice lacking the α₇ subunit. The results from this study suggest that the α₇ nAChR does not play an integral role in the acute toxicosis of MLA.

The purpose of this study was to investigate the toxic effect of long-term and low-level exposure to phorate using a metabonomics approach. Male Wistar rats were given phorate daily in drinking water at low doses of 0.05, 0.15 or 0.45 mg kg^–1 body weight (BW) for 24 weeks consecutively. The control group was given an equivalent volume of drinking water. Compared with the control group, 12 metabolites were significantly changed in the phorate-treated groups. Diethylthiophosphate (DETP) was considered as a biomarker of exposure to phorate. Phorate can cause disturbances in the energy-related metabolism, and other tissues and cell damages.

Oxidative stress has been recognized as a potential mediator of cell death. Astrocytes play an active role in brain physiology responding to harmful stimuli by activating astrogliosis, which in turn has been associated either with survival or degenerative events. The characterization of the mechanistic actions exerted by different toxins in astrocytes is essential to understand the brain function and pathology. As age plays a critical role in degenerative processes, the aim of this study was to determine whether the administration.

A simple, accurate and sensitive gas chromatography-mass spectrometry (GC-MS) method has been developed and validated for analyzing the metabolic profile of fatty acids and simultaneous quantification of 18 fatty acids including non-esterified fatty acids (NEFA) and esterified fatty acids (EFA) in acetaminophen (APAP)-induced liver injured and control rats serum.

Along with the chemometrics analyses of principle component analysis (PCA) and partial least squares- discriminant analysis (PLS-DA), APAP-induced and control rats could be differentiated and NEFAs such as oleic acid (C18:1n9), linoleic acid (C18:2n6), docosahexaenoic acid (C22:6n3) and arachidonic acid (C20:4n6) were identified as the potential biomarkers of APAP-induced hepatic injury.

The novel combined study of GC-MS analysis and chemometric analysis based on the metabolic profile of fatty acids was a prospective technique for the assessment of chemical-induced hepatotoxicity.

The protective effect of (E)-2-benzylidene-4-phenyl-1,3-diselenole (BPD) on renal injury induced by cisplatin in rats was investigated. BPD ameliorated renal injury and reduced plasma markers altered by cisplatin. BPD attenuated the reduction of renal AA and GSH content induced by cisplatin. The decrease of GST activity, but not GPx and CAT activities, in rats exposed to cisplatin was reversed by BPD. BPD was effective in attenuating the inhibition of δ-aminolevulinic dehydratase in kidneys of rats exposed to cisplatin.

Ascorbic acid (AA) pre-treatment increased growth inhibition by emodin in AR-positive LNCaP, PC-3, HCT-15, MG-63 and A549 cancer cells, all with varying LRP1 levels, probably owing to ROS generation by emodin and the pro-oxidant activity of AA. Marked increase in LRP1 expression and growth inhibition of these cancer cells by emodin under hypoxia-like conditions may point to a strategy to target hypoxic tumors through the LRP1 protein by increased uptake of emodin, alone or along with other chemotherapeutic agents.

The effect of chronic ingestion of strontium 90 was investigated in a mouse model, with strontium 90 concentrations representative of environmental contamination after a major nuclear accident. Results did not show evidence for modifications in the hematopoietic system. By contrast, a modification of bone metabolism was demonstrated, but was not linked to phosphocalcic homeostasis. This suggests a direct effect of strontium 90 storage in the bones on osteoblasts and/or osteoclasts.

This study investigated the genotoxicity of gold nanoparticles (AuNPs), silver nanoparticles (AgNPs), zinc oxide nanoparticles (ZnO NPs), and titanium dioxide nanoparticles (TiO₂ NPs) by using the SOS chromotest with E. coli PQ37. The four NPs and its ions, in a range of tested concentrations, exerted no effects in the SOS chromotest. They are classified as non-genotoxic on the basis of the SOS chromotest.

Previously, we demonstrated that maternal exposure to phthalates enhances atopic dermatitis in male mouse offspring. However, whether phthalate exposure affects neuroimmune biomarkers in allergic mice has not yet been studied. Di-(2-ethylhexyl) phthalate (DEHP) and di-isononyl phthalate (DINP) are environmental chemicals that are commonly used as plasticizers. This study was designed to investigate the expression levels of neuroimmune biomarkers in the hypothalamus of a murine model of allergic asthma after phthalate exposure throughout juvenility until adulthood.

Thirteen-week toxicity of (AA) was evaluated in Syrian hamsters, at concentrations of 0 (control), 20, 30 and 50 mg kg^–1 body weight in drinking water. Treatment with AA caused peripheral nerve disorders in all AA-treated groups with dose dependence. In addition, testicular toxicity at 50 mg kg^–1 and lowered body weights at 30 and 50 mg kg^–1 were observed. These results indicated the maximum tolerated dose for long-term studies of AA to be 20 mg kg^–1 or less in Syrian hamsters.

The occurrence of oocytes in the testis (testis-ova) is one reproductive disorder and can be used as a valid endpoint when studying disruptive effects of estrogenic chemicals in fish and amphibians. Gonads of the genetically male tropical clawed frog (Silurana tropicalis) exposed developmentally to 17α-ethinylestradiol were analyzed using histopathology, microarray and quantitative PCR. Expression of zpc and 42Sp50 genes was demonstrated to be associated with testis-ova in the S. tropicalis.

Osteoclasts (OCLs) are multinucleated bone-resorbing cells. In this study, we investigated the effects of tert-butylhydroquinone (tBHQ) on differentiation of macrophages into OCLs. tBHQ treatment decreased the expression of nuclear factor of activated T cells cytoplasmic-1 (NFATc1) and impaired phosphorylation of several mitogen-activated protein kinases. And, tBHQ inhibited the release of high mobility group box 1 (HMGB1), a recently identified activator of OCL differentiation. Thus, tBHQ inhibits OCL differentiation through the HO-1/HMGB1/NFATc1 pathways.

The aim of this study was to determine the effects of FZD on HepG2 cells by activating and inhibiting p38 MAPK signaling pathway. The cell cycle and proliferation of HepG2 cells treated with FZD were detected by flow cytometry and MTT assay in the presence or absence of p38 MAPK inhibitors (SB203580), respectively. Cyclin D1, cyclin D3, and CDK6 were detected by quantitative real-time PCR and western blot analysis. Our data showed that p38 MAPK became phosphorylated after stimulation with FZD. Activation of p38 MAPK could arise S-phase cell-cycle arrest and suppress cell proliferation. Simultaneously, inhibition of p38 MAPK significantly prevented S-phase cell-cycle arrest, increased the percentage of cell viability, and decreased the expression of cyclin D1, cyclin D3, CDK6. These results demonstrated that FZD arose S-phase cell-cycle arrest via activating p38 MAPK signaling pathway in HepG2 cells. Cyclin D1, cyclin D3, and CDK6 are target genes functioning at the downstream of p38 MAPK in HepG2 cells induced by FZD.

Thyroid hormones are essential for the regulation of a wide range of biological processes associated with normal development and metabolism in vertebrates. For the screening of chemicals having potential thyroid hormone and anti-thyroid hormone activities, we have established transient transactivation assay systems. We demonstrate that this in vitro transactivation system can be used for screening chemicals with potential thyroid hormone agonistic and antagonistic activities.

Formaldehyde is ubiquitous in the environment. It is a genotoxic substance. In this study, we investigated the effects of antioxidant on formaldehyde-induced genotoxicity in A549 Cell Lines. Formaldehyde caused induction of DPCs, increased MDA levels, and decreased SOD and GSH-Px activity. Moreover, the activation of NF-κB and AP-1 were induced. Curcumin is an important antioxidant. Pretreatment with curcumin could counteracted above effects. These results suggest that formaldehyde-induced genotoxicity induce through its ROS and lipid peroxidation and cause DPCs effects.

4-Methylesculetin (4-ME) is a potent synthetic antioxidant compound. This study investigated the in vivo genotoxicity of 4-ME and its potential antigenotoxicity against doxorubicin (DX) DNA damage. Doses of 500, 1000 and 2000 mg kg^–1 were tested only or with a simultaneous injection of DX. A comet assay in different cells of mice and the micronucleus test were the endpoints analyzed. No differences were observed between the control and treated groups, indicating that it lacks genotoxic and cytotoxic effects. Moreover, 4-ME demonstrated protective effects against DX.

The separate effects of Cd and Zn were studied on Enchytraeus doerjesi at 15, 20 and 25 °C. Cd LC50/EC50 generally decreased with increasing temperature. Zn LC50/EC50 consistently increased with increasing temperature. This indicates that Cd toxicity to E. doerjesi increases with increasing temperature while Zn toxicity decreases. Increasing temperature may have differential effects on metal toxicity depending on the biological importance of the metal and the endpoints of interest. Further research on more ecologically relevant metal mixtures is needed.

Mycotoxin sorption efficacy of UPSN, a refined calcium montmorillonite clay, was examined in vitro and a rapid in vivo hydra bioassay was utilized to evaluate aflatoxin B₁ and fumonisin B₁ toxicity and confirm UPSN efficacy. UPSN sorbed both mycotoxins in vitro, although more FB1 bound at pH 2.0 than 6.5. Treatment with UPSN resulted in significant protection to mycotoxin-exposed hydra. Results indicate that this modified hydra bioassay can be used as an initial screen to predict toxin binding agent efficacy.

Because of their small size, robust structure and unique characteristics, carbon nanotubes (CNTs) are increasingly being used in a variety of biomedical applications, materials and products. As their use increases, so does the probability of their unintended release and human exposure. Therefore, it is important to establish their potential biodistribution and biopersistence to better understand the potential effects of their exposure to humans.

Compound VAM2-6, a 7-nitro-4-(5-piperidinopentyl)quinoxalin-2-one, has been shown previously to exhibit an in vitro trichomonacide efficacy of 100% at a concentration of 100 µg ml^–1. The present study was undertaken to evaluate its ability to induce DNA damage in the peripheral blood of mice using the comet and micronucleus assays. Cell viability was assessed by the fluorochrome-mediated viability test. VAM2-6 induced DNA damage in a dose-dependent manner and decreased cell viability 24 h after treatment.

The protective effects of exogenous CO via intraperitoneal injection were investigated in a murine model of acute liver damage induced by GalN/LPS. It showed that CO remarkably improved the survival rate of mice and led to attenuated hepatocellular damage. CO inhibits hepatic MDA contents and restores SOD and GSH, as well as reducing iNOS/NO production. Additionally, CO substantially inhibits hepatocyte apoptosis and reduces pro-inflammatory cytokines levels. Thus, the protective effects of CO are mainly dependent on its anti-oxidative, anti-inflammatory and anti-apoptotic properties.

The present study investigated the in vitro toxic microcystin-LR (MC-LR) effects on immune cells isolated from the blood of carp. Moreover, the effect of the toxin on the phagocytic activity of leukocytes and on actin and tubulin re-organization in phagocytic cells was studied. Incubation of the cells with MC-LR inhibited phagocytosis without affecting apoptosis or glutathione levels. The toxin induced a significant re-organization of the actin cytoskeleton in phagocytes. These results suggest that fish phagocytes and lymphocytes are targets for MC-LR.

We examined quercetin-mediated changes in the expression of genes associated with oxidative stress and antioxidant defense in A549 type II-like alveolar epithelial cells exposed to PQ. Quercetin was shown to attenuate PQ-induced oxidative damage in alveolar A549 cells through the activation of the transcription factor Nrf2, induction of its target HO-1 expression and other antioxidant-related genes, which ultimately improves cell viability by reducing cellular ROS and increasing cellular GSH. These combined results indicate that quercetin may be used to mitigate or minimize oxidative stress via reduction of oxygen radicals.

In this study, the cytotoxicity of TiO₂ nanoparticles was investigated by analyzing the changes in metabolite profiles both in mouse fibroblast (L929) cells and their corresponding culture media using gas chromatograph with a time-of-flight mass spectrometry (GC/TOFMS)-based metabolomic strategy. It was found that the major biochemical metabolism (carbohydrate metabolism) was suppressed in TiO₂ nanoparticle-treated L929 cells and their corresponding culture media. These results might account for the serious damage to energy metabolism in mitochondria and the increased cellular oxidation stress in TiO₂ nanoparticle-induced L929 cells.

Human endogenous retrovirus (HERV) envelope protein-coated, baculovirus vector-based HPV16L1 (AcHERV-HPV16L1) is a non-replicating DNA vaccine. AcHERV-HPV16L1 DNA vaccine did not cause significant changes in body weight. No hematological changes were observed on day 30 post-treatment. In a repeated-dose toxicity study, a transient decrease in the body weights was recovered by post-treatment day 20. Organ weights had normalized by day 30. A AcHERV-based HPV16L1 DNA vaccine demonstrated preclinical safety criteria in acute and range-finding toxicity studies.

Peroxisome proliferator-activated receptor-γ (PPAR-γ) agonists not only improve metabolic abnormalities of diabetes and consequent diabetic nephropathy, but they also protect against non-diabetic kidney disease in experimental models. Here, we investigated the effect of PPAR-γ agonist pioglitazone against acute renal injury on a cisplatin model in mice. Nephrotoxicity was induced by a single intraperitoneal (i.p.) injection of cisplatin (10 mg kg^–1). Pioglitazone was administered for six consecutive days in doses of 15 or 30 mg kg^–1 day^–1, per os (p.o.), starting 3 days before cisplatin injection.

Perfluorooctane sulfonate (PFOS) is a widespread environmental contaminant that is detected in the lung of mammalian. The mechanisms underlying PFOS-induced lung cytotoxicity remain unclear. We treated human lung cancer A549 cells with PFOS and the cellular apoptosis, mitochondrial membrane potential, intracellular ROS were determined. PFOS induced a dose-dependent increase in A549 cells toxicity via apoptosis pathway with obvious oxidative stress. N-acetylcysteine (NAC) as an antioxidant can block PFOS-induced ROS generation, mitochondrial membrane potential loss and cell apoptosis.

p53-proficient and -deficient gpt delta mice were given pentachlorophenol (PCP), phenobarbital (PhB), or piperonylbutoxide (PBO) for 13 weeks. PCP or PBO significantly increased 8-OHdG levels in liver DNA. PhB or PBO significantly elevated CYP 2B10 mRNA levels while NQO1 levels were significantly increased in PCP- or PBO-treated mice. They failed to increase gpt and red/gam gene mutations in the liver independently of p53. The action of oxidative stress by these chemicals is likely limited to tumor promotion.

Brominated flame retardants (BFRs) are widely used in consumer products. To evaluate the effects of BFRs on immune responses, we investigated whether BFRs affect phenotypes and the function of splenocytes and bone marrow-derived dendritic cells in vitro. Polybrominated diphenyl ethers, hexabromocyclododecane, and tetrabromobisphenol A were examined. The results demonstrate that some BFRs may stimulate immune cells. BFRs can induce or enhance immune/allergic responses by increasing antigen presentation-related molecule expression and interleukin-4 production.

Morphine neurotoxicity has been studied in NG108-15 cells after 24 h incubation with 0.1 and 10 μM concentrations of the opioid. Images compatible with apoptosis and necrosis could be observed directly and after staining with methylene blue, crystal violet and propidium iodide/4′,6-diamidino-2-phenylindole. Quantification of caspase 3/7 activity and DNA fragmentation confirmed a modest but significant proapoptotic effect of morphine. Co-incubation with 10 μM yohimbine prevented all these changes, thus extending previous findings of a yohimbine-sensitive, neurotoxic effect of morphine on NG108-15 cells.

We investigate whether diethyl phthalate, one of the potential endocrine disrupters, affect sirtuins (SIRT1 and SIRT2) and methyltranferases (DNMT1 and DNMT3a) on the apoptosis of PC12 cells, because diethyl phthalate was reported previously to enhance apoptosis induced by serum deprivation. We found that DNMT3a was significantly decreased by serum deprivation. However, DNMT3a, DNMT3b and SIRT1were significantly increased under the enhancement of apoptosis induced by serum deprivation. These results suggest that SIRT1, DNMT3a and DNMT3b play multiple and complex roles in different apoptotic stages.

Ivermectin (IVM)- and Ivomec® (IVO)-induced DNA damage kinetics was analyzed by SCGE in CHO-K1 treated with 50 µg ml^–1 for 80 min. Viability remained unchanged up to 3 h. Afterwards, survival decreased up to 79% at 6 h, remaining unchanged within 12-24 h. An increase of induced damage was observed within 0-3 h whereas a decrease was revealed during the final 18 h. Both compounds induced apoptosis. The decrease in DNA lesions is mostly related to cytotoxicity than repair.

Multi-walled carbon nanotubes (MWCNTs) were intratracheally instilled as a single dose at 0.2 or 1.0 mg kg^–1 or a repeated dose at 0.04 or 0.2 mg kg^–1, once a week for 5 weeks, to male rats. Inflammatory changes were found in the lungs of rats after a single instillation and repeated instillation at both doses. MWCNTs had no potential for DNA damage in comet assays using the lung cells of rats given MWCNTs at doses causing inflammatory responses.

Sulfolane is a widely used industrial solvent that is often used for gas treatment, in the manufacture of polymers and electronics, and may be found in pharmaceuticals as a residual solvent. The lowest, most conservative, RfD of 0.01 mg kg⁻¹ per day was based on reduced white blood cell counts in female rats. This RfD was used to develop a tap water screening level that is protective of ingestion, viz. 365 µg l⁻¹.

Cirrhosis was induced by chronic TAA administration and the effects of co-administration of conventional coffee or decaffeinated coffee respectively for 8 weeks were evaluated. TAA administration elevated serum alkaline phosphatase, γ-glutamyl transpeptidase and alanine aminotransferase, liver lipid peroxidation and collagen content. Additionally increased levels of a number of proteins were detected including transforming growth factor-β, connective tissue growth factor and alpha-smooth muscle actin, and matrix metalloproteinase-2, 9 and 13. Coffee suppressed most of the changes produced by TAA.

Male mice were exposed to raw water from Meiliang Bay, Lake Taihu, China for 90 days. Hepatic transcriptomic profiles were determined. Compared with control group, raw water altered expression of 170 hepatic genes. Nine biological processes and nine biological pathways were significantly perturbed, mainly linked to the regulation of cell processes, DNA repair, chromatin modification, oxidative reduction and carbohydrate metabolism. This study provided excellent insights into early toxicological effects related to raw Lake Taihu water.

We developed a rodent newborn behavioral test method for evaluating the risk of neurotoxicity of chemicals, by means of determining the newborn's activity using the tare function of an analytical balance, and confirmed the usefulness of our developed testing method by determining the activity of mouse newborns with microcephaly induced by prenatal exposure to a neurotoxicant, methylnitrosourea.

16-20 month old C57BL/6 mice were exposed orally to 5.0 mg methylmercury per kg body weight. Methylmercury-treated aged mice performed significantly worse in open field, foot print analysis and a vertical pole test. Isolated cerebellar granule cells from methylmercury-treated aged mice exhibited higher reactive oxygen species levels and reduced mitochondrial membrane potentials. Aged mice exposed to methylmercury exhibited no greater impairment compared to young adult mice exposed to the same dose, as reported in earlier studies from this lab.

In this paper we combine network reconstruction with classification of unseen compounds to their appropriate toxicity class. We select gene pairs that are linked by a common expression pattern in one class and differ in this expression pattern in another class. The resulting gene dysregulation network provides novel and well-distinguishing pairs of markers. Moreover, paired expressions by their specific pattern generate new hypotheses on regulatory origin and/or common functionality of genes in the pairs, and how these differ between classes.

There is an increasing belief that organophosphorus compounds (OPCs) impair glucose homeostasis and cause hyperglycemia and diabetes mellitus. The present study was undertaken to investigate the putative diabetogenic effect of sub-lethal and sub-chronic exposure of paraoxon, an extremely hazardous OPC used in pesticides. The effect of paraoxon on streptozotocin-induced diabetic rats was also examined. The results suggest that paraoxon neither induces nor exacerbates diabetes in experimental rats.

Caffeine has various biological effects. Effect of (100 μM) caffeine on heartbeat, survival and expression of cell damage-related genes were studied. Hatching decreased significantly in caffeine treatment at 60 and 72 hpf. Heartbeat increased significantly at 72 and 96 hpf. Significant up-regulation was found in all tested genes, Bax/Bcl2 ratio, and significant down-regulation in Bcl2. Caffeine exposure, increase heartbeats, stimulates oxidative stress and may trigger apoptosis via a mitochondrial-dependent pathway. Use of caffeine harm the human fetus by affecting the cell-damage related genes.

A randomized, double-blind, placebo-controlled, crossover study in 20 healthy, human volunteers, aged 19–28 years, was carried out to refine the Acute Reference Doses (ARfD) of an atropine/scopolamine mixture. Bradycardia was identified as a critical effect. We estimate that the No Observed Adverse Effect Levels (NOAELs) for the atropine/scopolamine mixture are between the dose levels 0.12/0.10 and 0.37/0.29 µg kg^–1 body mass (BM). By applying the uncertainty factor of 10, we propose a new provisional ARfD of the mixture, i.e. 0.01 µg kg^–1 BM for each alkaloid.

Acute and chronic (or sub-chronic) toxicity of five selected antimicrobial agents, triclosan, triclocarban, resorcinol, phenoxyethanol and p-thymol, was investigated using green algae, daphnia and fish. Having compared the predicted no effect concentration (PNEC) determined from the toxicity data with measured environmental concentrations (MEC), a preliminary ecological risk assessment of these antimicrobials was conducted. The MEC/PNEC ratios were >1 for triclosan and triclocarban for some sites but were <0.1 for the other compounds in most cases.

Proprietary and reference molecules were used to compare the GADD45α-GFP ‘GreenScreen HC’ assay (GS) with in vitro screening tests. The GS renders the SOS-ChromoTest obsolete, confirms all results of the mini-Ames test and shows concordance of 82% with the micronucleus assay. The GS detected accurately catecholamines, carbendazim, tetracycline antibiotics and carbaryl. The GS produced negative results for metronidazole, dexamethasone, for rosiglitazone and for clindamycin and neomycin. The GS appears reproducible, robust, specific and sensitive test for genotoxicity screening.

Toxicogenomics was applied to develop a good model for the carcinogenicity prediction.

Impaired folate metabolism is considered a risk factor for neural tube defects (NTDs), but the relationship between folate dysmetabolism and NTDs remains unclear. The objective of this study was to develop a NTD model, and study the mechanism of NTDs. Methotrexate (MTX), a folate antagonist, induced NTDs in mice, and dihydrofolate reductase (DHFR) activity and folates derivatives concentrations of embryonic tissues decreased significantly. Furthermore, three high-confidence copy number variants (CNVs) were found on chromosome X, which in neural tube tissues may contribute to the development of NTDs.

Lead (Pb) is a testicular toxicant. In the present study, we investigated the effects of maternal Pb exposure during lactation on testicular development and steroidogenesis in male offspring. Maternal mice were exposed to different concentration of lead acetate (200 or 2000 ppm) through drinking water from postnatal day (PND) 0 to PND21. Results showed that Pb could be transported from dams to pups through milk. Maternal Pb exposure during lactation persistently disrupted testicular development and steroidogenesis in male offspring.

A metabolomics approach using proton nuclear magnetic resonance (NMR) was applied to investigate metabolic alterations following adriamycin (ADR) treatment for gastric adenocarcinoma. After BALB/c-nu/nu mice were implanted with human gastric adenocarcinoma, ADR (1 or 3 mg kg⁻¹ per day) was intraperitoneally administered for 5 days. Urinary endogenous metabolites related to the TCA cycle might be considered as therapeutic targets to potentiate the efficacy of ADR.

Single-walled carbon nanotubes (SWCNTs) had no mutagenicity in S. typhimurium TA98, TA100, TA1535, or TA1537, or in E. coli WP2uvrA, in the absence or presence of metabolic activation. SWCNTs did not increase the number of structural or numerical chromosomal aberrations after short-term or continuous exposure. In the micronucleus test using CD-1 mice, SWCNTs did not affect the proportion of immature erythrocytes, the total proportion of erythrocytes, or the number of micronuclei in immature erythrocytes.

The chemopreventive effect of geraniol was investigated against 7,12-dimethylbenz[a]anthracene (DMBA)/12-O-tetradecanoylphorbol 13-acetate (TPA)-mediated skin tumorigenesis in female Swiss albino mice. Effects of geraniol on TPA-induced skin edema, hyperplasia, COX-2, oxidative stress, ornithine decarboxylase and [3H] thymidine incorporation were observed. We studied geraniol treatment on incidence and number of tumor and on the expressions of Ras, Raf, ERK1/2, Bcl2 and Bax proteins in skin tumors. Our study concludes that Geraniol alters Ras proliferation pathway and pro-apoptotic state to inhibit DMBA/TPA-mediated skin tumorigenesis.

The cadmium (Cd) effect on ovary δ-aminolevulinate dehydratase (δ-ALA-D) activity was investigated in vitro and ex vivo. Cadmium inhibited cow ovary δ-ALA-D activity and seleno-furanoside did not reverse the Cd toxicity in vitro. Acute Cd exposure caused a significant inhibition on ovary δ-ALA-D activity in mice and seleno-furanoside was able in restoring enzyme activity. Ovarian δ-ALA-D activity is inhibited by Cd both in vitro and ex vivo and the seleno-furanoside therapy was effective after Cd acute exposure in mice.

Developmental effects of synthetic estrogen, diethylstilbestrol (DES), on mouse placentas were investigated by oral administration during early to mid-stage of gestation. Dose-dependent embryo mortality was observed. Exposure to DES impaired the development of the labyrinth and induced poor rough-surfaced endoplasmic reticulum in trophoblast giant cells. The results suggest that developmental changes induced by DES may be related to interference with nutrition and oxygen exchange between mother and fetus or a decrease in protein synthesis, resulting in fetal mortality.

BALB/c mice were injected every 3 days with various doses of CdSe/CdS-MPA QDs. Body weight of the mice treated with 25 mg/kg QDs was significantly decreased from day 7. The QDs were observed in the liver, spleen, lung, and kidneys, but not in brain, or heart. Significantly higher levels of LDH and NADPH oxidase were found in plasma, liver, and spleen. Histopathological examination did not show any tissue toxicity but IL-6 levels were increased in the plasma, liver, and spleen.

To understand the relationship between arsenic exposure and epigenetic modifications, in the present study we investigated the effects of long-term exposure to arsenic by focusing on the tumor-related gene expression and gene-specific epigenetic modulations in C57Bl/6 mice. The results of this study indicate that long-term arsenic exposure down-regulates p16^INK4a without altering DNA methylation in the normal liver, and suggest that its down-regulation is as a result of histone H3K9me2 by G9a recruitment.

The mycotoxin Zearalenone (ZEN) has been associated with a variety of adverse health effects. In order to better understood the mechanism of ZEN toxicity, a proteomic approach, based on the isotope approach iTRAQ, was applied to characterize cellular responses of HepG2 cells to ZEN exposure. This study identified about 982 proteins among which identification and quantification by MALDI-TOF (MS). IPA software was then used to determine the biological functions and canonical pathways associated with the ZEN-responsive proteins.

The hepatotoxicity effect of Coleus forskohlii root extract (CFE) standardized to 10% forskolin was tested in male ICR mice. Body and organ weights, food intake, plasma marker enzymes for liver damage and liver tissues histology were recorded. Both dietary CFE dose and time dependently affect the observed parameters whereas forskolin, the principle constituent in CFE, is not responsible for such effects.

Occupational and accidental exposure to polychlorinated biphenyls (PCBs) has been associated with an increased risk for lung cancer. Our results shown that, the serum levels of three PCB congeners with potential estrogenic activity were higher in lung cancer patients, which could have dual biological effects on lung tissue, not only inducing cell proliferation in non-neoplastic and neoplastic lung cells via estrogen receptor beta, but also increasing endogenous catechol levels.

The presence of pharmaceuticals and personal-care products (PPCPs) in aquatic environments is of concern. The aim of the present study was to use the RTG-2 cell line to analyse toxic effects of PPCPs present in wastewater-treatment-plant (WWTPs) effluents and their mixtures. The present study clearly shows that the stress response through which cells mount a homeostatic response to toxicants can be potentially used for an initial, rapid and cost-effective assessment of the complex mixtures of PPCP present in WWTP effluents.

The enantioselective effect of bifenthrin on antioxidant enzyme gene expression and stress protein response in PC12 cells were investigated. The results showed that exposure to 1S-cis-BF resulted in increased transcription of HSP90, HSP70, HSP60, Cu-Zn-SOD, Mn-SOD, CAT and GST, while exposure to 1R-cis-BF and rac-cis-BF exhibited these effects to lesser degrees. Induction of antioxidant enzyme gene expression produced by 1S-cis-BF might occur through activation of p38 MAPK and ERK, while increase of stress protein response might occur through p38 MAPK.

In three participating laboratories, photochemical reactivities of 42 coded chemicals and two standard controls were assessed by reactive oxygen species (ROS) assay for photosafety evaluation. Outcomes from the validation study were indicative of satisfactory transferability, intra- and inter-laboratory variability, and predictive capacity of the ROS assay. Thus, results from the present validation study provided sufficient support for the ROS assay as an alternative method for photosafety assessment.

Oleanolic acid (OA), a pentacyclic triterpenoid, exhibits potential anti-tumor activity against many tumor cell lines. This study aims to examine the anti-tumor activity of OA on pancreatic cancer cells and its potential molecular mechanism. The results showed that the proliferation of Panc-28 cells was inhibited by OA in a concentration-dependent manner, with an IC₅₀ (The half maximal inhibitory concentration) value of 46.35 µg ml⁻¹, as determined by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.

The present study was undertaken to evaluate the genotoxic potential of Copaifera langsdorffii leaf hydroalcoholic extract (CLE) and its influence on the genotoxicity induced by the chemotherapeutic agent doxorubicin (DXR) using the Swiss mouse peripheral blood micronucleus test. The results demonstrated that CLE itself was not genotoxic. In animals treated with CLE and DXR, the number of micronucleus was significantly decreased compared to animals receiving DXR alone.

The reversible AChE-inhibitors pyridostigmine, metoclopramide, tiapride, ranitidine, physostigmine, tacrine, amiloride, methylene blue, 7-methoxytacrine and K-27 were tested as prophylactic agents before exposure to the irreversible AChE-inhibitor paraoxon. Their efficacy in reducing organophosphorus compound-induced mortality was assessed by Cox survival analysis. Best in vivo protection from paraoxon-induced mortality was achieved by physostigmine or the oxime K-27, the most efficacious pretreatment agent not passing the blood brain barrier.

We previously reported that V⁵⁺ was able to decrease the TCDD-mediated induction of Cyp1a1 and Nqo1 gene expression levels in Hepa 1c1c7 and HepG2 cells; however, little is known regarding the in vivo effect. Thus, the objective of the current study was to investigate whether or not similar effects would occur at the in vivo level. Therefore, we examined the effect of co-exposure to V⁵⁺ and TCDD on the AhR-regulated genes. Our results demonstrated that V⁵⁺ modulates TCDD-induced AhR-regulated genes.

We previously demonstrated that exposure to very low concentrations of the organophosphorus insecticide chlorpyrifos (CPF) decreased electrical resistance across the blood–brain barrier (BBB) in vitro, indicating a loss of BBB integrity. This study examined the transient effects of CPF on expression of genes contributing to tight junctions of the BBB. The results suggest that altering gene expression for claudin5, transient receptor potential (canonical) channels (TRPC4), and zona occludens (ZO1) by CPF may directly contribute to BBB disruption, and that the alteration is reversible upon removal of CPF.

Microcystins (MCs) are highly liver-specific and evidenced as a liver tumor promoter. Oxidative stress is one of the most important toxicity mechanisms of MCs, which is tightly related to oxygen concentration. The effects of MCs on animals and cell lines in normoxia and the mechanisms have been well studied, but such effects in different oxygen conditions were still unclear.

The present study was performed to assess the toxicity of ¹⁸⁸Reliposome in Sprague-Dawley rats. None of the rats died and no clinical sign was observed, and only male rats administered 185 MBq of ¹⁸⁸Re-liposome displayed a weight loss. Although the WBC counts of both high-dose and medium-dose groups reduced 7-days postinjection, they recovered to a normal range before termination. There was no difference in urinary analyzes, biochemical parameters and histopathological assessments between the ¹⁸⁸Re-liposome-treated and control groups.

In mice lacking Mrp1, renal phase II and III enzyme expression was significantly changed owing to the loss of androgens, while in the lungs, phase II and phase III enzymes were downregulated in Mrp1^−/− mice, irrespective of their hormonal status. Expression in the small intestine was only different between intact and castrated Mrp1^−/− mice, and not different in wildtype FVB mice. Enzyme expression patterns correlated to Nrf2 expression. This information will aid in the understanding of how drug uptake, disposition and elimination can be influenced by both hormone status and the presence and magnitude of transporter expression.

In vivo geno- and antigenotoxicity of watercress juice were evaluated using comet assay (blood cells) and micronucleus test (bone marrow). Biopsies of bladder, epididymis and testicles of mice were performed to extend the experimental design. Watercress did not induce genetic damage per se and exhibited protective activity. Analysis of histological changes suggests a probable protective effect. Further studies are needed to establish the protective role of watercress juice against DNA damage.

Owing to the development of new materials and technology, the pollutants in the environment are becoming more varied and complex over time. In our previous study using ICR mice, we suggested that a single intratracheal instillation of single-walled carbon nanotubes (SWCNTs) induced early lung fibrosis and subchronic tissue damage. In the present study, to investigate the role of CCR5 in inflammatory responses to the uptake of SWCNTs, we compared BAL (Bronchoalveolar lavage) cell composition, cell cycles, cytokines, cell phenotypes, inflammatory response-related proteins, cell surface receptors and histopathology using CCR5 knockout (KO) and wild-type mice.

Nocodazole, paclitaxel and the antifungal griseofulvin, with a promising role in cancer treatment, affect microtubule dynamics by different ways. To compare their aneugenic properties we studied: (i) MN induction using CREST analysis, (ii) disturbance of mitotic spindle organization by immunofluorescence, and (iii) alterations in the expression of chromosome segregation regulating protein. We found that the generation of different types of abnormal metaphases dowing to centrosome deffects, as well as the alteration in the expression of proteins regulating chromosome segregation, was dependent on the drug and the cell line treated, reflecting their different effects on microtubule dynamics.

Pyrazinamide (PZA) is an important sterilizing prodrug that shortens the duration of tuberculosis therapy. However, hepatotoxicity has been reported during clinical trials investigating PZA. To determine the hepatotoxic effects of PZA in vivo and to further investigate the underlying cellular mechanism, we profiled the gene expression patterns of PZA-treated rat livers by microarray analysis. Wistar rats of both sexes were orally administered PZA at doses of 0.5, 1.0 and 2.0 g kg⁻¹ for 28 days.

Tissue factor (TF) is a membranous glycoprotein that functions as a receptor for coagulation factor VII/VIIa and activates the coagulation system when blood vessels or tissues are damaged. TF was upregulated in our monocrotaline (MCT)/lipopolysaccharide (LPS) hepatotoxicity model. We tested the hypothesis that TF-dependent fibrin deposition and lipid peroxidation in the form of oxidized low-density-lipoprotein (ox-LDL) accumulation contribute to liver inflammation induced by MCT/LPS in mice. In the present study, we blocked TF using antisense oligodeoxynucleotides against mouse TF (TF-ASO).

Microcystins (MCs) can cause evident hepatic apoptosis. In vitro studies indicated that uptake of MC by isolated hepatocytes was dramatically reduced as ambient temperature dropped, and some studies presented a hypothesis that differences in core body temperatures in animals result in diverse uptake of MC, as well as different toxic effects. Thus far, however, few in vivo studies have been conducted to investigate the effects of temperature on MC-induced hepatocyte apoptosis in fish, a typical poikilotherm.

A simple, rapid and sensitive GC/MS method has been developed and validated for the sensitive determination of cocaine and its main metabolites cocaethylene and benzoylecgonine in hair. The method is reproducible, robust and precise and fully satisfactory for its application to routine laboratory testing. The procedure was further applied to 40 hair samples from self-reported cocaine users. A new criterion was raised that contributes to minimize false-negative results and allows for a better interpretation of hair testing results.

The impact of ethanol on the body turnover of cadmium in a rat model reflecting excessive alcohol consumption in humans chronically exposed to moderate and relatively high levels of this heavy metal was studied. The results provide strong evidence that ethanol consumption under long-term moderate and relatively high exposure to cadmium modifies its metabolism in the organism, resulting in lower body burden of this toxic metal. Based on the findings, it can be concluded that cadmium concentration in the blood and tissues of alcohol abusers chronically exposed to moderate and relatively high levels of this metal may be lower, whereas its urinary excretion is higher than in their non-drinking counterparts. However, since ethanol is toxic itself, the decreased body burden of cadmium owing to alcohol consumption does not allow for the conclusion that the risk of health damage may be lower with co-exposure to these xenobiotics. In further study, it will be investigated how the ethanol-induced changes in the body status of cadmium influence the effects of its toxic action.

Methyl-tertiary butyl ether (MTBE) was shown to be anti-angiogenic in zebrafish, mice implanted with an ECGS fortified Matrigel plug and interfered with rat brain endothelial cells microcapillary tube formation. MTBE (500–1500 mg/kg) did not cause any growth or histological malformations in Fisher 344 dams or pups. MTBE is antiagiogenic, minimally toxic and possibly could be used to treat solid tumors.

In the present study, zebrafish embryos and larvae were used to investigate the sublethal and lethal effects of three different organochlorine pesticides, namely methoxychlor, endosulfan and heptachlor, as well as the flame retardant tetrabromobisphenol A, and its precursor compound bisphenol A. The use of vtg1 mRNA induction in zebrafish embryos and larvae was found to be a sensitive biomarker of exposure to these organic compounds, and was helpful in elucidating their adverse effects and setting water quality guidelines.

Although n-hexane is toxic to the ovary, the related mechanism remains unknown. Apoptosis of rat ovarian granulosa cells is thought to be one of the important reasons for ovarian toxicity. Rat ovarian granulosa cells were exposed in vitro to different concentrations of 2,5-hexanedione (HD, a major metabolite of n-hexane; 0, 20, 40 and 60 mmol l⁻¹) to observe the apoptosis, and its relevant gene expression was investigated. It was found by MTT assay that different exposure doses and different exposure time could inhibit the viability of ovarian granulosa cells.

A method is presented for the determination of drugs of abuse in vitreous humor using high-performance liquid chromatography and solid-phase extraction. A linear response from the detector was obtained within the concentration range of 0.1–4 µg mL⁻¹, the limits of detection were lower than 30 ng mL⁻¹, the coefficients of variation fluctuated between 0.1% and 12.4%, and the average recoveries were >78% for all the drugs except for EDDP, with a value of 66.4%.

The study of combined effects of pesticides represents a challenge for toxicology. In the case of the new growing generation of genetically modified (GM) plants with stacked traits, glyphosate-based herbicides (like Roundup) residues are present in the Roundup-tolerant edible plants (especially corns) and mixed with modified Bt insecticidal toxins that are produced by the GM plants themselves. The potential side effects of these combined pesticides on human cells are investigated in this work.

The IKr (hERG) current inhibition is considered to be the main target during the drug development process, although other ionic currents modification can either potentiate or mask hERG channel blockade. Database describing the results of in vitro studies investigating the chemical-IKs current interactions has been developed. Based on the collected data the predictive extended-QSAR models for the IC50 estimation were developed with the use of various algorithms. The best performing model was further built into the ToxComp platform (ToxIVIVE).

In this study, Table 1 shows the information of primer, anneal temperature and cycle of RT-PCR. PCR of all genes was performed under the conditions in this table. β-Actin was chosen as house-keeping gene. Table 2 is the results of apoptosis assay by flow cytometry. In the presence of dioscin, the rate of early apoptosis, advanced stage apoptosis and necrosis of cancer cells was increased in a dose-dependent manner.

Lead-induced liver damage and malfunction are partly due to a disturbance of the cellular antioxidant balance. Paraoxonase 1 (PON1) and PON2 are highly expressed in the liver and have been proposed as antioxidative enzymes. The effects of lead on PON1 and PON2 activities were investigated in human hepatoma HepG2 cells. Lead reduced PON2, but not PON1, activity and that this reduction was reversed by calcium. Lead-induced oxidative stress and decreased PON2 activity lead to the upregulation of PON2 transcript.

Cisplatin inhibited antioxidant enzymes (GST, GPx, GR, SOD and CAT) to a level that is significantly lower than the control group. Meanwhile treatment of cells with cisplatin in the presence of emodin significantly ameliorated the inhibited antioxidant enzymes. Controls contained all the reaction reagents except emodin or cisplatin.

We investigated the morphological and functional changes of testes in mice treated with a low dose of cadmium chloride (3mg CdCl₂/kg body weight) and also examined its toxicity as to susceptibility to EAO testicular autoimmunity. The results showed that exposure to 3 mg CdCl₂/kg body weight did not affect the spermatogenic state. However, the blood-testis barrier BTB at the tubuli recti and the rete testis, but not the seminiferous tubules, was slightly weakened, and intra-testicular mRNA expression of interleukin (IL)-6, tumor necrosis factor-α and IL-1β was significantly increased by the CdCl₂ treatment. Furthermore, immunization with testicular antigens after the CdCl₂ exposure significantly augmented the EAO severity. Therefore, exposure to a low dose of CdCl₂ induces no significant disturbance of spermatogenesis, however, it does the CdCl₂ treatment change the immunological microcircumstances in the testis, resulting in increased susceptibility to testicular autoimmunity.

Sensitisation of the respiratory tract by chemicals associated with occupational asthma is an important occupational health problem. Here the relationships between skin sensitising chemicals and those that preferentially or exclusively cause respiratory sensitisation is explored. It is demonstrated that chemical respiratory allergens test positive in standard methods for the identification of skin sensitising chemicals, and that this might provide important component of the hazard identification process for these chemicals.

Cytochrome P450-mediated desulfuration of organophosphorus compounds results in mechanism-based enzyme inhibition. Data suggest that reactive sulfur is released and binds to the apoprotein, although the identities of neither the adduct(s) nor the affected amino acid(s) have been clearly determined. In this work, 96 amu adducts to two cysteines of rat CYP3A1 were determined after incubation with methyl parathion. These adducts correspond to the mass of three sulfur atoms, as well as combinations of sulfur and oxygen.

Many different biomarkers can be used to evaluate ethanol intake. Ethyl glucuronide (EtG) is a minor metabolite of ethanol. Its investigation is of interest in both clinical and forensic contexts because of the wide window of detection. The analysis was performed by LC-ESI-MS/MS and SPE was used as a pretreatment. The method has been developed and fully validated according to the guidelines of forensic toxicology for the analysis of EtG in hair.

Jatropha oil is an emerging feedstock for the production of biodiesels. The increasing use of this non-edible, toxic oil will result in higher potential for accidental exposures. A repeated dose 28-day oral toxicity study was conducted to provide data for risk assessment. It is concluded that Jatropha oil produces adverse effects on female rats starting at 50 mg/kg/day with decreased white blood cell and lymphocyte counts and at 500 mg/kg/day in both genders in term of depressed growth rates.

A human in vivo toxicokinetic model was built to allow a better understanding of the toxicokinetics of folpet fungicide and its key biomarkers of exposure, and to simulate the transformation of folpet into phthalimide and other ring-metabolites: phthalamic and phthalic acids. The model closely reproduced the time courses of phthalimide in blood and urine as well as total ring-metabolites in urine of five volunteers administered orally 1 mg kg⁻¹ and dermally 10 mg kg⁻¹ of folpet.

Exposure of Wistar rats to MTBE in the drinking water for two years resulted in minimal exposure-related effects, including increased kidney weights. Chronic progressive nephropathy was evident in male and female rats and was exacerbated by exposure to MTBE. Brain astrocytomas were present but were not statistically significantly different from concurrent controls and fell within historical control ranges. Furthermore, the brain has not been identified as a target organ in previous chronic studies with MTBE or related compounds.

The adult male rats were treated with different doses of cypermethrin by gavage for consecutive 15 days. The weights of prostates and daily sperm production decreased. Seminiferous tubule changes included atrophy and distortion, reduction and deformation of spermatogonia and spermatocyte. Ultrastructural changes included disrupted cellular junctions, abnormal nucleus, necrosis of spermatogonia, spermatocytes and Sertoli cells. AR expression and serum testosterone reduced. Cypermethrin may impair the seminiferous tubules and spermatogenesis, which can be attributed to the reduced AR expression.

Hepatic effects of low-dose cadmium were examined in male Wistar rats given i.p. injection of Cd at 20 nmol kg⁻¹ body weight every other day for 4 weeks. At weeks 20, 44 and 52 after Cd treatment, the livers from Cd-treated and control rats were sampled. Mild histopathological changes, persistent oxidative damage and cell proliferation remained at weeks 44–52. These persistent changes may be associated with the persistent down-regulation of cellular antioxidant systems.

It is known that melphalan and chlorambucil exert aneugenic potential mediated through centrosome amplification. To further investigate their aneugenicity we: a) studied whether apoptosis is a mechanism for the elimination of damaged cells generated and b) investigated if chromosome segregation regulating proteins are involved in the modulation of their aneugenicity. We found that: a) apoptosis is not the mechanism of choice for selectively eliminating cells with supernumerary centrosomes and b) the proteins aurora-A, aurora-B and survivin are involved in the modulation of melphalan and chlorambucil aneugenicity.