RESUMO
Several hundred disinfection byproducts (DBPs) in drinking water have been identified, and are known to have potentially adverse health effects. There are toxicological data gaps for most DBPs, and the predictive method may provide an effective way to address this. The development of an in-silico model of toxicology endpoints of DBPs is rarely studied. The main aim of the present study is to develop predictive quantitative structure-activity relationship (QSAR) models for the reactive toxicities of 50 DBPs in the five bioassays of X-Microtox, GSH+, GSH-, DNA+ and DNA-. All-subset regression was used to select the optimal descriptors, and multiple linear-regression models were built. The developed QSAR models for five endpoints satisfied the internal and external validation criteria: coefficient of determination (R²) > 0.7, explained variance in leave-one-out prediction (Q²LOO) and in leave-many-out prediction (Q²LMO) > 0.6, variance explained in external prediction (Q²F1, Q²F2, and Q²F3) > 0.7, and concordance correlation coefficient (CCC) > 0.85. The application domains and the meaning of the selective descriptors for the QSAR models were discussed. The obtained QSAR models can be used in predicting the toxicities of the 50 DBPs.
Assuntos
Desinfecção/métodos , Água Potável/química , Modelos Moleculares , Compostos Orgânicos/toxicidade , Relação Quantitativa Estrutura-Atividade , Poluentes Químicos da Água/toxicidade , Simulação por Computador , Dicloroetilenos/química , Dicloroetilenos/toxicidade , Hidrocarbonetos Halogenados/química , Hidrocarbonetos Halogenados/toxicidade , Modelos Lineares , Cloreto de Metileno/química , Cloreto de Metileno/toxicidade , Estrutura Molecular , Compostos Orgânicos/química , Regressão Psicológica , Poluentes Químicos da Água/químicaRESUMO
Vinylidene chloride (VDC) has been widely used in the production of plastics and flame retardants. Exposure of B6C3F1 mice to VDC in the 2-year National Toxicology Program carcinogenicity bioassay resulted in a dose-dependent increases in renal cell hyperplasia, renal cell adenoma, and renal cell carcinomas (RCCs). Among those differentially expressed genes from controls and RCC of VDC-exposed mice, there was an overrepresentation of genes from pathways associated with chronic xenobiotic and oxidative stress as well as c-Myc overexpression and dysregulation of TP53 cell cycle checkpoint and DNA damage repair pathways in RCC. Trend analysis comparing RCC, VDC-exposed kidney, and chamber control kidney showed a conservation of pathway dysregulation in terms of overrepresentation of xenobiotic and oxidative stress, and DNA damage and cell cycle checkpoint pathways in both VDC-exposed kidney and RCC, suggesting that these mechanisms play a role in the pathogenesis of RCC in VDC-exposed mice.
Assuntos
Carcinoma de Células Renais , Dicloroetilenos/toxicidade , Neoplasias Renais , Estresse Oxidativo/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Proteína Supressora de Tumor p53/genética , Animais , Carcinoma de Células Renais/induzido quimicamente , Carcinoma de Células Renais/metabolismo , Carcinoma de Células Renais/patologia , Carcinoma de Células Renais/fisiopatologia , Relação Dose-Resposta a Droga , Feminino , Perfilação da Expressão Gênica , Rim/efeitos dos fármacos , Rim/patologia , Neoplasias Renais/induzido quimicamente , Neoplasias Renais/metabolismo , Neoplasias Renais/patologia , Neoplasias Renais/fisiopatologia , Masculino , Camundongos , Mutação , Testes de Toxicidade Crônica , Proteína Supressora de Tumor p53/metabolismoRESUMO
A majority (â¼80%) of human malignant mesotheliomas are asbestos-related. However, non-asbestos risk factors (radiation, chemicals, and genetic factors) account for up to 30% of cases. A recent 2-year National Toxicology Program carcinogenicity bioassay showed that male F344/N rats exposed to the industrial toxicant vinylidene chloride (VDC) resulted in a marked increase in malignant mesothelioma. Global gene expression profiles of these tumors were compared to spontaneous mesotheliomas and the F344/N rat mesothelial cell line (Fred-PE) in order to characterize the molecular features and chemical-specific profiles of mesothelioma in VDC-exposed rats. As expected, mesotheliomas from control and VDC-exposed rats shared pathways associated with tumorigenesis, including cellular and tissue development, organismal injury, embryonic development, inflammatory response, cell cycle regulation, and cellular growth and proliferation, while mesotheliomas from VDC-exposed rats alone showed overrepresentation of pathways associated with pro-inflammatory pathways and immune dysfunction such as the nuclear factor kappa-light-chain-enhancer of activated B cells signaling pathway, interleukin (IL)-8 and IL-12 signaling, interleukin responses, Fc receptor signaling, and natural killer and dendritic cells signaling, as well as overrepresentation of DNA damage and repair. These data suggest that a chronic, pro-inflammatory environment associated with VDC exposure may exacerbate disturbances in oncogene, growth factor, and cell cycle regulation, resulting in an increased incidence of mesothelioma.
Assuntos
Dicloroetilenos/toxicidade , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Doenças do Sistema Imunitário/induzido quimicamente , Inflamação/induzido quimicamente , Neoplasias Pulmonares/induzido quimicamente , Neoplasias Pulmonares/genética , Mesotelioma/induzido quimicamente , Mesotelioma/genética , Animais , Linhagem Celular Tumoral , Dano ao DNA , Feminino , Genes cdc/efeitos dos fármacos , Doenças do Sistema Imunitário/imunologia , Inflamação/fisiopatologia , Masculino , Mesotelioma Maligno , Análise em Microsséries , Neoplasias Peritoneais/induzido quimicamente , Neoplasias Peritoneais/patologia , RNA Neoplásico/biossíntese , Ratos , Ratos Endogâmicos F344 , Transdução de Sinais/efeitos dos fármacos , Neoplasias Testiculares/induzido quimicamente , Neoplasias Testiculares/patologiaRESUMO
Microbial dechlorination of trichloroethene (TCE) is inhibited at elevated TCE concentrations. A batch experiment and modeling analysis were performed to examine whether this self-inhibition is related to an enhanced cell decay or a reduced dechlorination activity at increasing TCE concentrations. The batch experiment combined four different initial TCE concentrations (1.4-3.0 mM) and three different inoculation densities (4.0 × 10(5) to 4.0 × 10(7)Geobacter cells·mL(-1)). Chlorinated ethene concentrations and Geobacter 16S rRNA gene copy numbers were measured. The time required for complete conversion of TCE to cis-DCE increased with increasing initial TCE concentration and decreasing inoculation density. Both an enhanced decay and a reduced activity model fitted the experimental results well, although the reduced activity model better described the lag phase and microbial decay in some treatments. In addition, the reduced activity model succeeded in predicting the reactivation of the dechlorination reaction in treatments in which the inhibiting TCE concentration was lowered after 80 days. In contrast, the enhanced decay model predicted a Geobacter cell density that was too low to allow recovery for these treatments. Conclusively, our results suggest that TCE self-inhibition is related to a reduced dechlorination activity rather than to an enhanced cell decay at elevated TCE concentrations.
Assuntos
Geobacter/citologia , Geobacter/efeitos dos fármacos , Halogenação/efeitos dos fármacos , Tricloroetileno/toxicidade , Técnicas de Cultura Celular por Lotes , Biodegradação Ambiental/efeitos dos fármacos , Dicloroetilenos/toxicidade , Dosagem de Genes , Geobacter/metabolismo , Modelos Biológicos , RNA Ribossômico 16S/genéticaRESUMO
A model that was used to describe toxicity from high concentrations of chlorinated aliphatic hydrocarbons (CAHs) on reductively dechlorinating cultures in batch reactors (Sabalowsky and Semprini (in press)) was extended here to simulate observations in continuous flow suspended and attached growth reactors. The reductively dechlorinating anaerobic Evanite subculture (EV-cDCE) was fed trichloroethene (TCE) and excess electron donor to accumulate cis-1,2-dichloroethene (cDCE) in a continuous flow stirred tank reactor (CFSTR); and an attached growth recirculating packed column (RPC). A concentration-dependent toxicity model used to simulate the results of batch reactors in part I (Sabalowsky and Semprini (in press) Biotechnol Bioeng) also simulated well the observations for the CFSTR and RPC growth modes. The toxicity model incorporates cDCE and TCE toxicity coefficients that directly increase the cell decay coefficient in proportion with cDCE and TCE concentrations. Simulated estimates of the cDCE and TCE toxicity coefficients indicate reductively dechlorinating cells are most sensitive to high concentrations of cDCE and TCE in batch-fed growth, followed by CFSTR, with attached growth being least sensitive. The greater toxicity of TCE than cDCE, and ratio of the modeled toxicity coefficients, agrees with previously proposed models relating toxicity to partitioning in the cell wall (K(M/B)), proportional to octanol-water partitioning (K(OW)) coefficients.
Assuntos
Reatores Biológicos/microbiologia , Chloroflexi/efeitos dos fármacos , Chloroflexi/metabolismo , Dicloroetilenos/metabolismo , Dicloroetilenos/toxicidade , Tricloroetileno/metabolismo , Tricloroetileno/toxicidade , Anaerobiose , Biotransformação , Chloroflexi/crescimento & desenvolvimento , OxirreduçãoRESUMO
A model was developed to describe toxicity from high concentrations of chlorinated aliphatic hydrocarbons (CAHs) on reductively dechlorinating cultures under batch-growth conditions. A reductively dechlorinating anaerobic Evanite subculture (EV-cDCE) was fed trichloroethene (TCE) and excess electron donor to accumulate cis-1,2-dichloroethene (cDCE) in batch-fed reactors. A second Point Mugu (PM) culture was also studied in the cDCE accumulating batch-fed experiment, as well as in a time- and concentration-dependent cDCE exposure experiment. Both cultures accumulated cDCE to concentrations ranging from 9,000 to 12,000 microM before cDCE production from TCE ceased. Exposure to approximately 3,000 and 6,000 microM cDCE concentrations for 5 days during continuous TCE dechlorination exhibited greater loss in activity proportional to both time and concentration of exposure than simple endogenous decay. Various inhibition models were analyzed for the two cultures, including the previously proposed Haldane inhibition model and a maximum threshold inhibition model, but neither adequately fit all experimental observations. A concentration-dependent toxicity model is proposed, which simulated all the experimental observations well. The toxicity model incorporates CAH toxicity terms that directly increase the cell decay coefficient in proportion with CAH concentrations. We also consider previously proposed models relating toxicity to partitioning in the cell wall (K(M/B)), proportional to octanol-water partitioning (K(OW)) coefficients. A reanalysis of previously reported modeling of batch tests using the Haldane model of Yu and Semprini, could be fit equally well using the toxicity model presented here, combined with toxicity proportioned to cell wall partitioning. A companion paper extends the experimental analysis and our modeling approach to a completely mixed reactor and a fixed film reactor.
Assuntos
Chloroflexi/efeitos dos fármacos , Chloroflexi/metabolismo , Dicloroetilenos/metabolismo , Dicloroetilenos/toxicidade , Tricloroetileno/metabolismo , Tricloroetileno/toxicidade , Anaerobiose , Biotransformação , Chloroflexi/crescimento & desenvolvimento , Modelos Biológicos , Modelos Teóricos , Oxirredução , Fatores de TempoRESUMO
The objective of this investigation was to elucidate the effects of route of exposure and oral dosage regimen on the toxicokinetics (TK) of 1,1-dichloroethylene (DCE). Fasted male Sprague-Dawley rats that inhaled 100 or 300 ppm for 2 h absorbed total systemic doses of (10 or 30 mg/kg DCE, respectively. Other groups of rats received 10 or 30 mg/kg DCE by intravenous injection, bolus gavage (by mouth), or gastric infusion (g.i.) over a 2-h period. Serial microblood samples were taken from the cannulated, unanesthetized animals and analyzed for DCE content by gas chromatography to obtain concentration versus time profiles. Inhalation resulted in substantially higher peak blood concentrations and area under blood-concentration time curves (AUC(0)(2)) than did gastric infusion of the same dose over the same time frame at each dosage level, although inhalation (AUC(0)(infinity)) values were only modestly higher. Urinary N-acetyl-beta-D-glucosaminidase (NAG) and gamma-glutamyltranspeptidase (GGT) activities were monitored as indices of kidney injury in the high-dose groups. NAG and GGT excretion were much more pronounced after inhalation than gastric infusion. Administration of DCE by gavage also produced much higher Cmax and AUC(0)(2) values than did 2-h g.i., although AUC(0)(infinity) values were not very different. The 30 mg/kg bolus dose produced marked elevation in serum sorbitol dehydrogenase, an index of hepatocellular injury. Administration of this dose by inhalation and gastric infusion was only marginally hepatotoxic. These findings demonstrate the TK and target organ toxicity of DCE vary substantially between different exposure routes, as well as dosage regimens, making direct extrapolations untenable in health risk assessments.
Assuntos
Dicloroetilenos/toxicidade , Acetilglucosaminidase/metabolismo , Administração por Inalação , Administração Oral , Animais , Dicloroetilenos/administração & dosagem , Dicloroetilenos/farmacocinética , Dicloroetilenos/farmacologia , Relação Dose-Resposta a Droga , Injeções Intravenosas , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Masculino , Ratos , Ratos Sprague-Dawley , Fenômenos Fisiológicos Respiratórios/efeitos dos fármacos , Transglutaminases/metabolismoRESUMO
OBJECTIVE: To explore the effects of p,p'-DDE and beta-BHC on the apoptosis of Sertoli cells in vitro via activation of Caspase. METHODS: Sertoli cells were treated in vitro for 24 hours with a serial concentrations of p,p'-DDE (10, 30 and 50 micromol/L), beta-BHC (10, 30 and 50 micromol/L) and p,p'-DDE + beta-BHC (10, 30 and 50 micromol/L). The inhibitory group was first treated with 100 micromol/L Caspase-3 inhibitor Ac-DEVD-CHO treating for 2 hours before 50 micromol/L p, p'-DDE + 50 micromol/L beta-BHC 24 hours-treatment. The vitality of Sertoli cells was determined by MTT and the apoptosis rate was measured by AO/EB double fluorescence staining. The expressions of Caspase-3, Caspase-8 and Caspase-9 were determined by RT-PCR. RESULTS: Average optical density (A) values were 0.498 +/- 0.039, 0.481 +/- 0.065, 0.397 +/- 0.032 and 0.286 +/- 0.049 in p,p'-DDE groups (10, 30, 50 and 70 micromol/L), and 0.518 +/- 0.103, 0.490 +/- 0.060, 0.454 +/- 0.054 and 0.302 +/- 0.030 in beta-BHC groups (10, 30, 50 and 70 micromol/L). In the mixture-treated groups (10, 30 and 50 micromol/L), the average A values were 0.483 +/- 0.048, 0.473 +/- 0.058 and 0.337 +/- 0.052. Compared with the solvent control group (0.527 +/- 0.022) , 50 micromol/L group of p, p'-DDE, beta-BHC or their mixture caused a significant decrease of Sertoli cell viability (t values were 4.599, 2.716, 6.537 respectively, P < 0.05). AO/EB double fluorescence staining analysis showed that apoptosis rates of Sertoli cells were significantly increased with all treated groups. The expressions of Caspase-3, Caspase-8 and Caspase-9 were upregulated as the concentrations of p,p'-DDE, beta-BHC and their mixture were increased. CONCLUSION: p,p'-DDE, beta-BHC and their mixture could induce the apoptosis of Sertoli cells in vitro which was associated with activation of Caspase-3 mediated by cleavage of Caspase-8 and Caspase-9.
Assuntos
Apoptose/efeitos dos fármacos , Dicloroetilenos/toxicidade , Hexaclorocicloexano/toxicidade , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/metabolismo , Animais , Caspase 3/metabolismo , Caspase 8/metabolismo , Caspase 9/metabolismo , Células Cultivadas , Masculino , RNA Mensageiro/genética , Ratos , Ratos Sprague-DawleyRESUMO
Occupational allergy and asthma is a challenging issue in the developing countries. Chemicals inhaled in the workplaces may act not only as allergens but also as immune response modifiers, contributing to asthma exacerbation. In this study, we tested the adjuvant effect of 20 ppm chloroform, 10 ppm 1,1-dichloroethylene, and 100 ppm styrene in mice. Female BALB/c mice were sensitised to ovalbumin (OVA) without using alum. During the OVA-sensitisation period, these mice were exposed by inhalation to the chemicals studied for 6h/day for four consecutive days. After two OVA-intratracheal challenges, a mild Th2 immune response was observed in the OVA-exposed groups. This response was characterised by a mild increase in serum specific IgE level, in local Th2 cytokine production, and in lung inflammatory reaction. Exposure to styrene or chloroform alone slightly increased Th2 cytokine production by lung-draining lymph node cells cultured with concanavaline A, except for the IL-4 level in the chloroform exposure group, which decreased. On the other hand, exposure to 1,1-dichloroethylene alone markedly increased the Th2 cytokine levels compared to those observed in the groups exposed to OVA alone. In the combined OVA+chemical-treated groups, styrene potentiated IL-4, -5 and -13 production efficiently (approximately two, four and three times higher, respectively), resulting in an increase in the total IgE levels and inflammatory reaction. On the other hand, the enhanced IgE levels and the exacerbation of the inflammatory response by 1,1-dichloroethylene or chloroform were associated with only minor changes in local cytokine levels. These findings suggest that exposure to chemicals through inhalation may aggravate the allergic lung inflammation. And this, depending on the chemical exposure conditions, may result from the synergistic effect of chemicals and allergen on local Th2 cytokine production.
Assuntos
Inflamação/patologia , Exposição por Inalação/efeitos adversos , Ovalbumina/imunologia , Hipersensibilidade Respiratória/patologia , Compostos de Alúmen/toxicidade , Animais , Asma/patologia , Clorofórmio/administração & dosagem , Clorofórmio/toxicidade , Citocinas/metabolismo , Dicloroetilenos/administração & dosagem , Dicloroetilenos/toxicidade , Ensaio de Imunoadsorção Enzimática , Eosinófilos/imunologia , Eosinófilos/metabolismo , Feminino , Imunoglobulina E/metabolismo , Interleucinas/metabolismo , Pulmão/patologia , Linfonodos/citologia , Camundongos , Camundongos Endogâmicos BALB C , Solventes/administração & dosagem , Solventes/toxicidade , Estireno/administração & dosagem , Estireno/toxicidade , Células Th2/efeitos dos fármacos , Células Th2/imunologiaRESUMO
Trichloroethylene (TCE) and dichloroethylene (DCE) are high-volume industrial chemicals frequently found as contaminants in public drinking water supplies. The developmental toxicity of both chemicals has been evaluated in laboratory and epidemiologic studies. It has been suggested that TCE and DCE are specific cardiac teratogens and that drinking water contaminated with them increases the risk of congenital heart defects in exposed human populations. In contrast, other laboratory and epidemiologic studies do not find an increase in developmental effects, either in general or specifically affecting the heart. This laboratory and epidemiologic base was reviewed to evaluate the strengths and weaknesses of the conflicting published reports. We conclude that the weight of experimental and epidemiologic evidence does not support the hypothesis that TCE or DCE is a selective developmental toxicant in general or a cardiac teratogen specifically.
Assuntos
Anormalidades Induzidas por Medicamentos/etiologia , Dicloroetilenos/toxicidade , Exposição Ambiental/efeitos adversos , Solventes/toxicidade , Teratogênicos/toxicidade , Tricloroetileno/toxicidade , Poluentes Químicos da Água/toxicidade , Adulto , Animais , Animais de Laboratório , Dicloroetilenos/classificação , Feminino , Humanos , Gravidez , Teratogênicos/classificação , Tricloroetileno/classificaçãoRESUMO
1,1-Dichloroethylene (DCE) causes pulmonary injury that is characterized by necrosis of bronchiolar Clara cells. Mitochondria have been identified as an early target in the toxic response. Because mitochondria have been implicated in both necrotic and apoptotic cell death, we have undertaken studies to test the hypothesis that DCE induces apoptosis, in addition to necrosis, in murine lung. A primary objective is to identify the biochemical events associated with pulmonary apoptosis. Groups of female CD-1 mice were treated with DCE (75 mg/kg i.p.) or corn oil. Using an antibody directed against DCE-cysteine conjugates, adducts were detected primarily in association with mitochondria in the apices of bronchiolar Clara cells. Furthermore, morphological studies demonstrated early mitochondrial alterations in Clara cells that included severe swelling and disruption of cristae. Western blotting of lung cytosolic proteins showed greater immunoreactivity for cytochrome c in fractions from mice treated with DCE for 4 h than in controls. Immunohistochemical studies with an antibody to activated caspase-3 and terminal deoxynucleotidyl transferase dUTP nick-end labeling were used to detect apoptotic cells. In both experiments, positive reactivities were observed in the bronchiolar epithelium at 12 and 24 h after DCE treatment, whereas reactivities were absent in tissues from control animals. Finally, bronchiolar epithelial cells showing morphological criteria of apoptosis (chromatin condensation and margination) were observed at 24 h after 75 and 125 mg/kg DCE. Apoptotic-like cells were more abundant in larger bronchioles. These data suggested that DCE produces pulmonary bronchiolar apoptosis by inducing mitochondrial perturbations, causing release of cytochrome c into the cytosol and caspase activation.
Assuntos
Apoptose/efeitos dos fármacos , Brônquios/citologia , Dicloroetilenos/toxicidade , Pulmão/citologia , Mitocôndrias/efeitos dos fármacos , Animais , Brônquios/patologia , Brônquios/ultraestrutura , Caspase 3 , Caspases/metabolismo , Citocromos c/metabolismo , Ativação Enzimática/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Células Epiteliais/ultraestrutura , Feminino , Immunoblotting , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Pulmão/patologia , Pulmão/ultraestrutura , Camundongos , Microscopia Eletrônica , Mitocôndrias/patologia , Mitocôndrias/ultraestruturaRESUMO
Experience with dose response and mechanisms of toxicity has shown that multiple mechanisms may exist for a single agent along the continuum of the full dose-response curve. It is highly likely that critical, limiting steps in any given mechanistic pathway may become overwhelmed with increasing exposures, signaling the emergence of new modalities of toxic tissue injury at these higher doses. Therefore, dose-dependent transitions in principal mechanisms of toxicity may occur, and could have significant impact on the interpretation of reference data sets for risk assessment. To illustrate the existence of dose-dependent transitions in mechanisms of toxicity, a group of academic, government, and industry scientists, formed under the leadership of the ILSI Health and Environmental Sciences Institute (HESI), developed a series of case studies. These case studies included acetaminophen, butadiene, ethylene glycol, formaldehyde, manganese, methylene chloride, peroxisome proliferator-activated receptor (PPAR), progesterone/hydroxyflutamide, propylene oxide, vinyl acetate, vinyl chloride, vinylidene chloride, and zinc. The case studies formed the basis for technical discourse at two scientific workshops in 2003.
Assuntos
Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/induzido quimicamente , Flutamida/análogos & derivados , Acetaminofen/administração & dosagem , Acetaminofen/farmacocinética , Acetaminofen/toxicidade , Analgésicos não Narcóticos/administração & dosagem , Analgésicos não Narcóticos/farmacocinética , Analgésicos não Narcóticos/toxicidade , Antagonistas de Androgênios/administração & dosagem , Antagonistas de Androgênios/farmacocinética , Antagonistas de Androgênios/toxicidade , Animais , Butadienos/administração & dosagem , Butadienos/farmacocinética , Butadienos/toxicidade , Dicloroetilenos/administração & dosagem , Dicloroetilenos/farmacocinética , Dicloroetilenos/toxicidade , Relação Dose-Resposta a Droga , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/metabolismo , Compostos de Epóxi/administração & dosagem , Compostos de Epóxi/farmacocinética , Compostos de Epóxi/toxicidade , Etilenoglicol/administração & dosagem , Etilenoglicol/farmacocinética , Etilenoglicol/toxicidade , Flutamida/administração & dosagem , Flutamida/farmacocinética , Flutamida/toxicidade , Formaldeído/administração & dosagem , Formaldeído/farmacocinética , Formaldeído/toxicidade , Humanos , Manganês/administração & dosagem , Manganês/farmacocinética , Intoxicação por Manganês/metabolismo , Cloreto de Metileno/administração & dosagem , Cloreto de Metileno/farmacocinética , Cloreto de Metileno/toxicidade , Oxirredução , Receptores Ativados por Proliferador de Peroxissomo/fisiologia , Progesterona/administração & dosagem , Progesterona/farmacocinética , Progesterona/toxicidade , Compostos de Vinila/administração & dosagem , Compostos de Vinila/farmacocinética , Compostos de Vinila/toxicidade , Zinco/administração & dosagem , Zinco/farmacocinética , Zinco/toxicidadeRESUMO
Tetrachloroethylene (PCE), a dry cleaning and degreasing solvent, can enter groundwater through accidental leaks or spills, and concentrations as high as 75 mg/L have been reported in Canadian aquifers. Amphibians in wetlands receiving contaminated groundwater may be exposed to PCE and its degradation products, but little information is available on the impacts of these compounds on indigenous amphibian species. Acute (96-h static renewal) exposures to PCE and its major degradation products, trichloroethylene (TCE) and cisand trans-dichloroethylene, were conducted on embryos of four North American amphibian species: wood frogs (Rana sylvatica), green frogs (R. clamitans), American toads (Bufo americanus), and spotted salamanders (Ambystoma maculatum). Subsequently, chronic exposures to PCE and TCE were conducted with the larvae of American toads. Both PCE and TCE were teratogenic to amphibian embryos; median effective concentrations (EC50s) for developmental deformities produced by PCE and TCE exposure for wood frogs and green frogs were 12 and 40 mg/L, respectively. Embryonic survivorship, however, was not compromised at these concentrations. American toads were less sensitive; the EC50 for developmental abnormalities was not attained at the highest test concentrations, 45 and 85 mg/L PCE and TCE, respectively. These results are pertinent in assessing the impact of groundwater pollution on an aquifer-fed wetland.
Assuntos
Anfíbios/embriologia , Anfíbios/crescimento & desenvolvimento , Dicloroetilenos/toxicidade , Exposição Ambiental , Solventes/toxicidade , Tetracloroetileno/toxicidade , Tricloroetileno/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Anormalidades Congênitas/etiologia , Anormalidades Congênitas/veterinária , Larva/crescimento & desenvolvimento , Testes de ToxicidadeRESUMO
1,1-Dichloroethylene (DCE) causes dysfunction of hepatic mitochondria. As mitochondria have been implicated in apoptosis through opening of the permeability transition pore (PTP), we have undertaken studies to test the hypothesis that DCE induces apoptosis, in addition to necrosis, in murine liver. Our primary objective was to identify the biochemical events associated with DCE-induced apoptosis. Female CD-1 mice were treated with a mildly hepatotoxic dose of DCE (125 mg/kg, i.p.). Using the fluorescent dye JC-1 (5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine iodide), decreased hepatic mitochondrial membrane potential was detected at 2 h. Western blotting of liver cytosolic proteins showed greater immunoreactivity for cytochrome c in fractions from mice treated with DCE for 4 h than in controls. Furthermore, caspase-9 activity was significantly increased 6 h after DCE exposure. Immunohistochemical studies with an antibody to activated caspase-3 and terminal deoxynucleotidyl transferase dUTP nick-end labeling staining were used to detect apoptotic cells. In both experiments, positive reactivities were observed in centrilobular hepatocytes 12 and 24 h after DCE. Additionally, centrilobular hepatocytes showing morphological criteria of apoptosis were observed at 24 h. Apoptosis and all apoptotic events were inhibited by pretreatment for 20 min with cyclosporine A (CyA) (50 mg/kg), a specific inhibitor of the mitochondrial PTP. To determine a major role for mitochondrial permeability transition (MPT) in DCE hepatotoxicity, serum alanine aminotransferase (ALT) activity was evaluated. ALT activity was significantly elevated 2 to 24 h after DCE, and CyA failed to inhibit this activity. These data suggested that DCE produces apoptosis by inducing MPT, causing release of cytochrome c into the cytosol and caspase activation.
Assuntos
Morte Celular/efeitos dos fármacos , Dicloroetilenos/toxicidade , Fígado/efeitos dos fármacos , Alanina Transaminase/metabolismo , Animais , Caspase 3 , Caspase 9 , Caspases/metabolismo , Citocromos c/análise , Ativação Enzimática , Feminino , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Fígado/patologia , Potenciais da Membrana/efeitos dos fármacos , Camundongos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/fisiologiaRESUMO
Using immunotoxic functional tests, namely IgM response to sheep red blood cells (SRBCs) and interferon-gamma (IFN-gamma) production, this study simultaneously evaluated the effects of inhaled chloroform (10, 20, and 50 ppm), carbon tetrachloride (100, 200, and 300 ppm), 1,1-dichloroethylene (5, 10, and 15 ppm), and styrene (100, 200, and 300 ppm) on the systemic (spleen) and local (lung-associated lymph nodes) immune response. At least one concentration of all the chemicals studied provoked a statistically significant increase in IgM response in the lymph nodes compared with the controls, as expressed by the number of plaque-forming cells (PFCs), whereas only the highest concentration of 1,1-dichloroethylene provoked an increase in the number of PFCs statistically different from the controls in the case of the spleens. The release of IFN-gamma in the lymph node cell cultures of the exposed mice exceeded that of the controls by more than 600%, whereas the release of IFN-gamma in the spleen cell cultures of the exposed mice was moderately different from the controls. It would appear from these results that the lung-associated lymph nodes are sensitive targets for chemical inhalation and that the results of systemic tests in the spleen may not mirror local immune response dysfunction. For risk assessment of inhaled chemicals, it is therefore important to take the local immunotoxic effects into consideration, in particular immunostimulation which may be involved in the rise in allergic diseases in industrialised countries.
Assuntos
Poluentes Ocupacionais do Ar/efeitos adversos , Imunidade/efeitos dos fármacos , Exposição por Inalação/efeitos adversos , Animais , Tetracloreto de Carbono/administração & dosagem , Tetracloreto de Carbono/toxicidade , Células Cultivadas , Clorofórmio/administração & dosagem , Clorofórmio/toxicidade , Dicloroetilenos/administração & dosagem , Dicloroetilenos/toxicidade , Eritrócitos/imunologia , Feminino , Imunoglobulina M/biossíntese , Imunoglobulina M/imunologia , Interferon gama/biossíntese , Pulmão/citologia , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Linfonodos/citologia , Linfonodos/efeitos dos fármacos , Linfonodos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Ovinos/imunologia , Solventes/administração & dosagem , Solventes/toxicidade , Baço/citologia , Baço/efeitos dos fármacos , Baço/imunologia , Estireno/administração & dosagem , Estireno/toxicidadeRESUMO
Hepatotoxicity induced by 1,1-dichloroethylene (DCE) is mediated by cytochrome P450-dependent metabolism to reactive intermediates, including the epoxide. We have tested the hypothesis that mitochondria are a primary target of toxicity by investigating dose- and time-dependent effects of DCE on mitochondrial respiration. Hepatotoxicity, as assessed by serum alanine aminotransferase (ALT) activity, was evaluated. We have also determined the effectiveness of N-acetyl-L-cysteine (NAC) in protecting against respiratory perturbations and hepatotoxicity. Liver mitochondria were isolated 2 h after DCE (50, 75, 100, 125, and 150 mg/kg) treatment. Glutamate (complex I)- and succinate (complex II)-supported mitochondrial respiration was assessed by measurement of state 3 (ADP-stimulated) and state 4 (resting) rates of oxygen consumption. The corresponding respiratory control ratios (RCRs, state 3/state 4) and ADP:O ratios were then calculated. A DCE dose of 125 mg/kg significantly inhibited glutamate- and succinate-supported state 3 respiration, leading to a significant reduction in corresponding RCRs and ADP:O ratios. In time-dependent studies, state 3 respiration rates and RCRs for glutamate-supported respiration were significantly decreased as early as 20 min after DCE (125 mg/kg) treatment, whereas those for succinate-supported respiration were significantly decreased at 90 min. Additionally, ADP:O ratios for glutamate-supported respiration were significantly decreased starting at 60 min, and those for succinate-supported respiration at 90 min. Alterations in mitochondrial function preceded significant increases in ALT activity, which was first manifested at 2 h. Pretreatment with NAC (1200 mg/kg) abrogated DCE-induced GSH depletion and inhibited disturbances in mitochondrial respiration. Moreover, NAC protected against increased ALT activity, suggesting that the protective effect of NAC is due to increased GSH for conjugation reactions and/or its antioxidant property. These results showed that DCE-mediated mitochondrial dysfunction is an early event that preceded the onset of hepatotoxicity.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/patologia , Dicloroetilenos/toxicidade , Mitocôndrias Hepáticas/patologia , Acetilcisteína/farmacologia , Difosfato de Adenosina/metabolismo , Alanina Transaminase/metabolismo , Animais , Relação Dose-Resposta a Droga , Feminino , Glutationa/metabolismo , Técnicas In Vitro , Indicadores e Reagentes , Camundongos , Mitocôndrias Hepáticas/efeitos dos fármacos , Consumo de Oxigênio/efeitos dos fármacos , Polarografia , Fatores de TempoRESUMO
1,2-Dichloroethylene exists in two isomeric states: trans-1,2-dichloroethylene and cis-1,2-dichloroethylene. The trans isomer is used more widely in industry than the cis isomer. trans-1,2-Dichloroethylene is used as a solvent for waxes, resins, and acetylcellulose. It is also used in the extraction of rubber, as a refrigerant, and in the manufacture of pharmaceuticals and artificial pearls. F344/N rats and B6C3F1 mice were administered trans-1,2-dichloroethylene in microcapsules in feed for 14 weeks. Animals were evaluated for clinical pathology, reproductive system effects, and histopathology. Genetic toxicity studies were conducted in vitro in Salmonella typhimurium and Chinese hamster ovary (CHO) cells, and in vivo in mouse bone marrow cells and peripheral blood erythrocytes. In the 14-week feed studies, groups of 10 male and 10 female rats and mice were fed diets containing microcapsules with a chemical load of 45% trans-1,2-dichloroethylene. Dietary concentrations of 3,125, 6,250, 12,500, 25,000, and 50,000 ppm microencapsulated trans-1,2-dichloroethylene resulted in average daily doses of 190, 380, 770, 1,540, and 3,210 mg/kg for male rats; 190, 395, 780, 1,580, and 3,245 mg/kg for female rats; 480, 920, 1,900, 3,850, and 8,065 mg/kg for male mice; and 450, 915, 1,830, 3,760, and 7,925 mg/kg for female mice. Additional groups of 10 male and 10 female rats and mice served as untreated and vehicle controls. There were no exposure-related deaths of rats or mice. Mean body weights of male rats and male and female mice in the 50,000 ppm groups were significantly less than those of the vehicle controls. The mean body weight gains of female mice in the 12,500 and 25,000 ppm groups were also significantly less than that of the vehicle controls. On day 21 and at week 14, there were mild decreases in hematocrit values, hemoglobin concentrations, and erythrocyte counts in groups of male and female rats in the 25,000 and 50,000 ppm groups. At week 14, these effects were seen in male rats exposed to 6,250 and 12,500 ppm. There were no exposure-related alterations in clinical chemistry parameters in rats or mice. The liver weights of female rats exposed to 6,250 ppm or greater were significantly greater than those of the vehicle controls. The absolute kidney weights of male rats exposed to 25,000 or 50,000 ppm were significantly decreased. No gross or microscopic lesions were observed in rats or mice that could be attributed to trans-1,2-dichloroethylene exposure. Neither cis-, trans-, nor cis,trans-1,2-dichloroethylene was mutagenic in S. typhimurium strain TA97 (cis isomer only), TA98, TA100, TA1535, or TA1537, with or without S9 metabolic activation enzymes. In CHO cells in vitro, cis- 1,2-dichloroethylene induced sister chromatid exchanges (SCEs) in the absence of S9; with S9, the single trial that was performed yielded equivocal results. The cis,trans isomer induced significant increases in SCEs in cultured CHO cells with and without S9. In contrast to these positive results, trans-1,2-dichloroethylene gave negative results in the SCE test, with and without S9. Neither cis-, trans-, nor cis,trans-1,2-dichloroethylene induced chromosomal aberrations (Abs) in cultured CHO cells, with or without S9. In vivo, no induction of SCEs or Abs was noted in bone marrow cells of male mice administered cis- or trans-1,2-dichloroethylene by intraperitoneal injection once, with sampling performed 23 hours (for SCE analyses) or 17 hours (for Abs analyses) after injection. In addition, negative results were obtained in a peripheral blood micronucleus test in male and female mice administered trans- 1,2-dichloroethylene in microcapsules in feed for 14 weeks. Very little toxicity was associated with ingestion of microencapsulated trans-1-2-dichloroethylene. Histopathology and clinical chemistry data, combined with body and organ weight data, revealed that the maximum tolerated dose was not reached in these studies.
Assuntos
Dicloroetilenos/toxicidade , Animais , Cápsulas , Testes de Carcinogenicidade , Dicloroetilenos/administração & dosagem , Dieta , Relação Dose-Resposta a Droga , Feminino , Rim/efeitos dos fármacos , Rim/patologia , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Dose Máxima Tolerável , Camundongos , Testes de Mutagenicidade , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Endogâmicos F344RESUMO
Inhalation studies were conducted to determine the potential subchronic toxicity of a mixture of trans-1,2-dichloroethylene (70%), cis-1,2-dichloroethylene (5%), and perfluorobutylethylene (25%). Groups of rats were exposed to 0, 400, 2000, or 8000 ppm concentrations of the mixture vapor 6 h/day, 5 days/wk, for a total of 20 exposures. Subgroups of rats were further observed during a 1-mo recovery period. Functional observational battery (FOB) and motor activity (MA) behavioral tests were conducted prior to initiation of the exposures, during exposure wk 4, and after a 1-mo postexposure recovery period. Clinical pathology evaluations were conducted at the end of the exposure period and after a 1-mo recovery period. At the end of the 4-wk exposure period, tissues from rats were collected, histologically processed, and evaluated by light microscopy. Test substance-related, biologically significant decreased body weights and body weight gains occurred in male and female rats exposed to 8000 ppm. In addition, test substance-related, statistically significant decreases in food consumption and/or food efficiency were observed in male rats exposed to 8000 ppm. During exposures to 8000 ppm, some rats exhibited tremors and ataxia. Usually tremors and ataxia were observed within 1 h after initiation of the daily exposure period and were observed during each exposure day. Tremors were also observed during 1 exposure day in the 2000 ppm animals. In addition to the tremors and ataxia, rats exposed to 2000 ppm or 8000 ppm had a diminished and/or no alerting response to a sharp, sound stimulus during each of the daily exposure periods. These effects were transient since no clinical observations of compromised neurological function were detected when the rats were evaluated upon return to the animal room following exposure. Daily reoccurrence of this apparently acute effect in the 8000 ppm group did not produce enduring neurological changes since there were no test substance-related effects on FOB parameters or on MA conducted the day following the last exposure or during the recovery period. In addition, there were no toxicologically significant changes in hematology, clinical chemistry, or urinalysis parameters in either males or females for any exposure concentration; and there were no test substance-related gross or microscopic morphological changes in males or females administered any exposure concentration. Under the conditions of the study, the no-observed-effect level (NOEL) was 400 ppm in males and females based on clinical signs of toxicity during exposure to 2000 or 8000 ppm.
Assuntos
Ataxia/induzido quimicamente , Dicloroetilenos/toxicidade , Hidrocarbonetos Fluorados/toxicidade , Exposição por Inalação , Animais , Dicloroetilenos/administração & dosagem , Relação Dose-Resposta a Droga , Ingestão de Alimentos/efeitos dos fármacos , Feminino , Fluorocarbonos , Hidrocarbonetos Fluorados/administração & dosagem , Masculino , Doenças do Sistema Nervoso/induzido quimicamente , Nível de Efeito Adverso não Observado , Ratos , Fatores Sexuais , Aumento de Peso , Redução de PesoRESUMO
The U.S. Environmental Protection Agency has classified 1,1-dichloroethylene (vinylidene chloride; VDC) as a "C" carcinogen and has developed an inhalation unit risk value and an oral cancer slope factor for this chemical. The development and use of these cancer potency estimates for risk assessment purposes are questionable. The inhalation unit risk value is based on increased kidney adenocarcinomas in Swiss mice from one study. This type of cancer was not increased in female mice or in rats or hamsters in the same study nor in male mice of a similar strain in another study with higher VDC exposures. The VDC oral cancer slope factor is based on a non-statistically significant increase in adrenal pheochromocytomas in male rats following oral exposure in a standard National Toxicology Program chronic bioassay. Both human and animal literature relevant to VDC carcinogenicity was reviewed according to the USEPA draft Guidelines for Carcinogen Risk Assessment with the objective of determining the weight-of-evidence for VDC carcinogenicity. We conclude that information currently available for VDC is most appropriately characterized in a weight-of-evidence narrative by the descriptor "inadequate for an assessment of human carcinogenic potential." For chemicals with this descriptor, dose-response assessment is not indicated. Under this guidance, quantitative estimates of cancer risks associated with VDC exposure are inappropriate until additional, more definitive evidence for human carcinogenicity becomes available.