RESUMEN
A rule-based expert system (ES) was developed to predict chemical binding to the estrogen receptor (ER) patterned on the research approaches championed by Gilman Veith to whom this article and journal issue are dedicated. The ERES was built to be mechanistically transparent and meet the needs of a specific application, i.e. predict for all chemicals within two well-defined inventories (industrial chemicals used as pesticide inerts and antimicrobial pesticides). These chemicals all lack structural features associated with high affinity binders and thus any binding should be low affinity. Similar to the high-quality fathead minnow database upon which Veith QSARs were built, the ERES was derived from what has been termed gold standard data, systematically collected in assays optimized to detect even low affinity binding and maximizing confidence in the negatives determinations. The resultant logic-based decision tree ERES, determined to be a robust model, contains seven major nodes with multiple effects-based chemicals categories within each. Predicted results are presented in the context of empirical data within local chemical structural groups facilitating informed decision-making. Even using optimized detection assays, the ERES applied to two inventories of >600 chemicals resulted in only ~5% of the chemicals predicted to bind ER.
Asunto(s)
Sistemas Especialistas , Sustancias Peligrosas/toxicidad , Relación Estructura-Actividad Cuantitativa , Antiinfecciosos/clasificación , Antiinfecciosos/toxicidad , Sustancias Peligrosas/clasificación , Plaguicidas/clasificación , Plaguicidas/toxicidad , Receptores de Estrógenos/metabolismo , Pruebas de Toxicidad/métodosRESUMEN
Regulatory agencies are charged with addressing the endocrine disrupting potential of large numbers of chemicals for which there is often little or no data on which to make decisions. Prioritizing the chemicals of greatest concern for further screening for potential hazard to humans and wildlife is an initial step in the process. This paper presents the collection of in vitro data using assays optimized to detect low affinity estrogen receptor (ER) binding chemicals and the use of that data to build effects-based chemical categories following QSAR approaches and principles pioneered by Gilman Veith and colleagues for application to environmental regulatory challenges. Effects-based chemical categories were built using these QSAR principles focused on the types of chemicals in the specific regulatory domain of concern, i.e. non-steroidal industrial chemicals, and based upon a mechanistic hypothesis of how these non-steroidal chemicals of seemingly dissimilar structure to 17ß-estradiol (E2) could interact with the ER via two distinct binding types. Chemicals were also tested to solubility thereby minimizing false negatives and providing confidence in determination of chemicals as inactive. The high-quality data collected in this manner were used to build an ER expert system for chemical prioritization described in a companion article in this journal.
Asunto(s)
Estrógenos/clasificación , Animales , Disruptores Endocrinos/química , Disruptores Endocrinos/clasificación , Disruptores Endocrinos/toxicidad , Estrógenos/toxicidad , Parabenos/química , Parabenos/clasificación , Parabenos/toxicidad , Fenoles/química , Fenoles/clasificación , Fenoles/toxicidad , Relación Estructura-Actividad Cuantitativa , Receptores de Estrógenos/metabolismo , Salicilatos/química , Salicilatos/clasificación , Salicilatos/toxicidad , TruchaRESUMEN
Hallmark signs of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) toxicity in rainbow trout sac fry, are yolk sac edema, hemorrhage, craniofacial malformation, and growth retardation culminating in mortality. Our objective was to determine the role of cardiovascular dysfunction in the development of this toxicity. An embryotoxic TCDD dose (385 pg/g egg) caused a progressive reduction in blood flow in rainbow trout sac fry manifested first and most dramatically in the 1st and 2nd branchial arches and vessels perfusing the lower jaw. Blood flow was reduced later in the infraorbital artery and occipital vein of the head as well as segmental vessels and caudal vein of the trunk. Reduced perfusion occurred last in gill branchial arteries involved with oxygen uptake and the subintestinal vein and vitelline vein involved with nutrient uptake. Although heart rate throughout sac fry development was not affected, heart size at 50 days post-fertilization (dpf) was reduced far more than body weight or length, suggesting that the progressive circulatory failure caused by TCDD is associated with reduced cardiac output. Craniofacial development was arrested near hatch, giving rise to craniofacial malformations in which the jaws and anterior nasal structures were underdeveloped. Unlike the medaka embryo, in which TCDD causes apoptosis in the medial yolk vein, endothelial cell death was not observed in rainbow trout sac fry. These findings suggest a primary role for arrested heart development and reduced perfusion of tissues with blood in the early-life stage toxicity of TCDD in trout.
Asunto(s)
Sistema Cardiovascular/efectos de los fármacos , Anomalías Craneofaciales/inducido químicamente , Oncorhynchus mykiss/fisiología , Dibenzodioxinas Policloradas/toxicidad , Teratógenos/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Vasos Sanguíneos/efectos de los fármacos , Volumen Cardíaco/efectos de los fármacos , Edema/patología , Embrión no Mamífero , Corazón/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Etiquetado Corte-Fin in Situ , Oncorhynchus mykiss/anomalías , Oncorhynchus mykiss/embriología , Flujo Sanguíneo Regional/efectos de los fármacos , Factores de Tiempo , Membrana Vitelina/irrigación sanguínea , Membrana Vitelina/efectos de los fármacos , CigotoRESUMEN
Mono-ortho PCBs are global contaminants of wildlife with the potential to produce toxicity by an aryl hydrocarbon receptor (AhR)-mediated mechanism. To determine the potency of 2,3,3',4, 4'-pentachlorobiphenyl (PCB 105) for producing reproductive and developmental toxicity, adult ring-necked pheasant hens (Phasianus colchicus) were orally dosed with 0, 0.06, 0.6, or 6 mg PCB 105/kg hen/week for 10 weeks to achieve cumulative doses of 0, 0.6, 6, or 60 mg PCB 105/kg hen after which hens were bred with untreated roosters once per week for 8 weeks. Except at week 6 of the egg-laying period when cumulative egg production in the 6 mg PCB 105/kg hen group was greater than controls, fertilized egg production was not significantly different between treatment groups. Embryo mortality and chick mortality were not significantly different between treatment groups. Total body and heart weights of all chicks 1 day posthatch (dph) were not different between groups, however, liver weights of chicks from the 60 mg/kg treatment group were greater than controls at 1 dph. The first chick to hatch from each hen was reared to 21 dph and among these birds, the total body, liver, and heart weights were not different between groups. There were no dose-related malformations of the beak or limbs, and no signs of subcutaneous edema, ascites, or pericardial edema in chicks at 1 or 21 dph. Hepatic microsomal monooxygenase activities [ethoxyresorufin-O-dealkylase (EROD), benzyloxyresorufin-O-dealkylase (BROD), and methyloxyresorufin-O-dealkylase (MROD)] were significantly elevated in chicks at 1 dph from hens given a cumulative PCB 105 dose of 6 mg/kg and in chicks at 21 dph from hens given a cumulative PCB dose of 60 mg/kg. These results indicate that a cumulative PCB 105 dose up to 60 mg/kg hen does not decrease the production of fertilized eggs or increase embryo or chick mortality in ring-necked pheasants, but does increase chick hepatic monooxygenase activity.
Asunto(s)
Aves/fisiología , Bifenilos Policlorados/toxicidad , Reproducción/efectos de los fármacos , Animales , Sistema Enzimático del Citocromo P-450/efectos de los fármacos , Sistema Enzimático del Citocromo P-450/metabolismo , Relación Dosis-Respuesta a Droga , Embrión no Mamífero/efectos de los fármacos , Desarrollo Embrionario , Óvulo/efectos de los fármacos , Óvulo/crecimiento & desarrolloRESUMEN
Toxicity and histopathology of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in zebrafish (Danio rerio) early life stages was characterized from 12 to 240 hr postfertilization (hpf) following water-borne exposure of newly fertilized eggs. TCDD did not increase egg mortality (0-48 hpf), nor did it affect time to hatching (48-96 hpf). Egg doses of 1.5 ng [3H]TCDD/g or greater elicited toxic responses in zebrafish larvae. Pericardial edema and craniofacial malformations were first observed at 72 hpf, followed by the onset of yolk sac edema (96 hpf) and mortality (132 hpf). At 240 hpf the ED50s for pericardial edema, yolk sac edema, and craniofacial malformations were 2.2, 2.1, and 1.9 ng [3H]TCDD/g egg, respectively. The LD50, determined at 240 hpf, was 2.5 ng [3H]TCDD/g egg. Severe hemodynamic changes, observed as slowed blood flow in vascular beds of the trunk, head, and gills and slowed heart rate, occurred in TCDD-treated zebrafish prior to or coincident with the onset of gross signs of toxicity. Histological examination of TCDD-treated zebrafish revealed a variety of epithelial tissue lesions including arrested gill development and ballooning degeneration and/or necrosis of the renal tubules, hepatocytes, pancreas, and all major brain regions. Mesenchymal tissue lesions included subcutaneous edema in the head, trunk, and yolk sac, edema of the pericardium and skeletal muscle, and underdevelopment of the swim bladder. This demonstration of zebrafish responsiveness to TCDD early life stage toxicity coupled with the considerable information on developmental biology and genetics of zebrafish provides a foundation for future investigations into the mechanism of TCDD developmental toxicity.
Asunto(s)
Anomalías Inducidas por Medicamentos/etiología , Dibenzodioxinas Policloradas/toxicidad , Pez Cebra/embriología , Anomalías Inducidas por Medicamentos/patología , Animales , Sistema Cardiovascular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Edema/inducido químicamente , Embrión no Mamífero/irrigación sanguínea , Embrión no Mamífero/efectos de los fármacos , Embrión no Mamífero/patología , Frecuencia Cardíaca/efectos de los fármacos , Flujo Sanguíneo Regional/efectos de los fármacos , Cráneo/anomalías , Cráneo/efectos de los fármacos , Cráneo/embriología , Especificidad de la Especie , Saco Vitelino/efectos de los fármacos , Saco Vitelino/patología , Pez Cebra/crecimiento & desarrollo , Cigoto/efectos de los fármacosRESUMEN
Polybrominated and polychlorinated biphenyls (PBBs/PCBs), dibenzo-p-dioxins (PBDDs/PCDDs), dibenzofurans (PBDFs/PCDFs), and diphenyl ethers (PBDEs/PCDEs) are persistent, lipophilic environmental contaminants that may pose a risk to fish early life stage survival. To determine this potential risk, a rainbow trout early life stage mortality bioassay was used in which the potency of individual polybrominated chemicals was compared to the potency of the most potent polychlorinated chemical in these classes, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Following injection of newly fertilized rainbow trout eggs, fish-specific toxic equivalency factors (TEFs) were calculated as the molar ratio of TCDD LD50 to brominated compound LD50. Signs of toxicity were identical to those produced by polychlorinated TCDD-like chemicals and included yolk sac edema, pericardial edema, multifocal hemorrhages, reduced growth, and craniofacial malformations. Polybrominated dibenzo-p-dioxins, dibenzofurans, and biphenyls exhibited decreased potency with increased bromine substitution. Only 2,3,7,8-TBDD was more potent than 2,3,7,8-TCDD, whereas other polybrominated dibenzo-p-dioxins were equipotent or less potent than identically substituted polychlorinated dibenzo-p-dioxins in this assay. Although two PBDF congeners were equipotent to identically substituted PCDFs, 2,3,7,8-TBDF was 9-fold more potent than 2,3,7,8-TCDF. Both 3,3',4,4'-TBB and 3,3',4,4',5,5'-HxBB were 10-fold more potent than identically substituted polychlorinated biphenyls. The halogenated diphenyl ethers and di-ortho polybrominated biphenyls were inactive in this assay. Thus, in this in vivo assay the polybrominated and polychlorinated TCDD-like chemicals were not always equally potent. To assess the risk posed by mixtures of these chemicals to feral fish populations, fish-specific TEFs for both polybrominated and polychlorinated chemicals should be used.
Asunto(s)
Benzofuranos/toxicidad , Compuestos de Bifenilo/toxicidad , Embrión no Mamífero/efectos de los fármacos , Hidrocarburos Bromados/toxicidad , Oncorhynchus mykiss/crecimiento & desarrollo , Óvulo/efectos de los fármacos , Éteres Fenílicos/toxicidad , Dibenzodioxinas Policloradas/toxicidad , Animales , Éteres Difenilos Halogenados , Dosificación Letal Mediana , Bifenilos Polibrominados/toxicidadRESUMEN
Use of fish-specific toxic equivalency factors (TEFs) to estimate the risk that exposure to polybrominated dibenzo-p-dioxins (PBDDs), dibenzofurans (PBDFs), and biphenyls (PBBs) pose to fish early life stage survival depends on validation of the hypothesis that these chemicals act additively to produce mortality. A rainbow trout early life stage bioassay was used to determine how pairs of PBDD, PBDF, and PBB congeners interact to produce 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-like toxicity associated with sac fry mortality. The congener pairs tested were 2,3,7,8-tetrabromodibenzo-p-dioxin (2,3,7,8-TBDD)/1,2,3,7,8-pentabromodibenzo-p-dioxin [correction of pentabromodibenzop-dioxin] (1,2,3,7,8-PBDD); 2,3,7,8-TBDD/1,2,3,7,8-pentabromodibenzofuran (1,2,3,7,8-PBDF); 1,2,3,7,8-PBDD/2,3,4,7,8-pentabromodibenzofuran (2,3,4,7,8-PBDF); and 2,3,4,7,8-PBDF/ 3,3',4,4'-tetrabromobiphenyl (3,3',4,4'-TBB). Graded doses of each congener alone, or graded doses of fixed ratios of paired congeners were injected into newly fertilized rainbow trout eggs. In all cases, interactions between congener pairs were additive as tested by a probit model. Isobolographic analysis also supported the hypothesis that the PBDD, PBDF, and PBB congeners act additively. Thus, the use of fish-specific TEFs to convert fish tissue concentrations of individual PBDD, PBDF, and PBB congeners to TCDD equivalents (TEs) and then adding the TEs contributed by the various congeners to give the total TCDD equivalents concentration (TEC) in the tissue is supported by these results. By comparing the TEC in feral fish eggs to the fish egg TCDD no-observed-effect level (NOEL) and lowest-observed-effect level (LOEL) for early life stage mortality, the risk that complex mixtures of these polybrominated chemicals in eggs pose to sac fry survival can be estimated.
Asunto(s)
Benzofuranos/toxicidad , Compuestos de Bifenilo/toxicidad , Embrión no Mamífero/efectos de los fármacos , Bifenilos Polibrominados/toxicidad , Dibenzodioxinas Policloradas/toxicidad , Animales , Sinergismo Farmacológico , Desarrollo Embrionario , Oncorhynchus mykiss , Análisis de SupervivenciaRESUMEN
Fish-specific toxic equivalency factors (TEFs), which relate the toxic potency of polyhalogenated aromatic hydrocarbons (PHAHs) to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) based on the endpoint of early life stage mortality, have been used in assessing the risk to fish early life stage survival of complex mixtures of PHAHs in feral fish eggs. Use of TEFs assumes that PHAH congeners act additively. However, this has not been unequivocally determined. Isobolograms and a probit model were used to assess the validity of the additivity assumption by determining the significance of interactions between pairs of polychlorinated dibenzo-p-dioxin (PCDD), dibenzofuran (PCDF), and biphenyl (PCB) congeners when injected into newly fertilized rainbow trout eggs in ratios bracketing those found in feral lake trout eggs from the Great Lakes. The majority of congener pairs tested acted additively in causing rainbow trout early life stage mortality: [1,2,3,7,8- pentachlorodibenzo-p-dioxin (1,2,3,7,8-PCDD)/TCDD]; [2,3,4,7,8-pentachlorodibenzofuran (2,3,4,7,8-PCDF)/1,2,3,7,8-PCDD]; (2,3,4,7,8-PCDF/TCDD), (2,3,7,8-tetrachlorodibenzofuran/2,3,4,7,8-PCDF); [3,3',4,4'-tetrachlorobiphenyl (PCB 77)/3,3',4,4',5-pentachlorobiphenyl (PCB 126)]; [2,3,3',4,4'-pentachlorobiphenyl (PCB 105)/TCDD]; (2,2',4,4',5,5'-hexachlorobiphenyl/TCDD); (PCB 105/PCB 126); and (2,3',4,4',5-pentachlorobiphenyl/PCB 126). The only pairs showing evidence of a statistically significant interaction that deviated from additivity were (TCDD/PCB 77) and (TCDD/PCB 126). Taken together, these results suggest that the use of fish-specific TEFs to determine TCDD equivalents contributed by individual congeners in a fish egg sample and then adding these TCDD equivalents to determine the total amount contributed by all congeners may not exactly predict the mortality risk posed to fish early life stages by the mixture of TCDD-like congeners in the eggs. However, the relatively small deviations from additivity in the rainbow trout sac fry mortality test (1- to 4-fold) are less than traditional uncertainty factors used in noncancer risk assessments (10-fold/factor) and are not sufficient to warrant a change away from the additivity assumption in assessing the risk to fish early life stage mortality posed by TCDD and related compounds in eggs.