RESUMEN
Exposure to crude oil and certain petroleum products can be a serious health hazard. Clarified slurry oil (CSO) is a complex mixture of hydrocarbons derived from the processing of crude oil, and is a known systemic and developmental toxicant, mutagen, and carcinogen. In the present study, CSO and two crude oils, Belridge heavy crude oil (BHCO) and Lost Hills light crude oil (LHLCO), were examined for their estrogenic and antiestrogenic properties in a human breast-cancer cell (MCF-7) assay. The MCF-7 focus assay is based on postconfluent cell growth and tissue restructuring, measured as the postconfluent development of multicellular nodules or foci. The mutagenicity indices of BHCO and LHLCO also were determined in a modified Ames Salmonella assay. Oil samples were prepared in dimethyl sulfoxide, resulting in extraction of virtually all of the aromatic compounds including the sulfur- and nitrogen-substituted three- to seven-ring polycyclic aromatic compounds comprising 62.2% of the CSO, 9% of the BHCO, and 2% of the LHLCO by total weight. None of the three samples was estrogenic in the MCF-7 focus assay. In contrast, all of the samples were antiestrogenic; that is, they inhibited the development of foci induced by 1 nM 17beta-estradiol (E2). The potencies of the oil samples for both antiestrogenicity and mutagenicity were correlated with the percent of polycyclic aromatic compounds they contained. Two potential mechanisms for the observed antiestrogenicity were examined. Radiometric analysis of the catabolism of [3H]E2 in MCF-7 cell cultures demonstrated that all three samples increased catabolism of E2. Results from a whole-cell estrogen-receptor (ER) binding assay suggested that metabolites of compounds in the oil samples might have competed with [3H]E2 for ER in the MCF-7 cultures. Thus the antiestrogenicity of the oil samples may occur through at least two mechanisms, increased catabolism of E2 and antagonistic binding to ER.
Asunto(s)
Neoplasias de la Mama/patología , Antagonistas de Estrógenos/farmacología , Petróleo/toxicidad , Unión Competitiva/efectos de los fármacos , Medios de Cultivo , Estradiol/sangre , Femenino , Humanos , Pruebas de Mutagenicidad , Receptores de Estrógenos/metabolismo , Salmonella/efectos de los fármacos , Salmonella/genética , Células Tumorales CultivadasRESUMEN
U.S. Navy submarines reported a yellowing of metal surfaces on their internal surfaces. The yellowing was initially identified on the painted steel bulkheads but further examination indicated that it was not limited to steel surfaces and included bedding, thread tape, Formica, plastisol covered hand-wheels, and aluminum lockers. Crew members also reported to the medical department that their skin turned yellow when they came in contact with these contaminated surfaces and requested information on the effects of exposure. Studies conducted by General Dynamics' Electric Boat Division (EBD) determined that the agent was 2,6-Di-tertbutyl-4-Nitrophenol (DBNP). 2,6-Di-butylphenol (DBP) is an antioxidant additive used in lubricating oils and hydraulic fluids. In the enclosed atmosphere of a submarine, the oil mist could be spread throughout the boat by venting the lube oil to the atmosphere. Submarines use electrostatic precipitators (ESP) to clean the air of particulate materials. During passage through the ESP, oil mist containing DBP is nitrated to DBNP, which is then moved throughout the boat in the ventilation system. Analysis of the EBD data indicated 24-hour exposure concentrations to be in the range of <3.0 to 122 ppb in the laboratory and submarine settings. Submarine crews may be exposed to these concentrations for as many as 24 hours/ day for 90 days during underway periods. Toxicity studies regarding the oral and dermal uptake of DBNP were conducted. From the literature the lethal dose to 50 percent of the population (LD50) of DBNP (rat) was reported by Vesselinovitch et al. in 1961 to be 500 mg/kg. Our studies indicated that the LD50 is in the range of 80 mg/kg in the rat. Our work also includes dermal absorption studies, which indicated that DBNP is not well absorbed through intact skin. Within this study, no no-observable adverse effect level (NOAEL) or lowest observable adverse effect level (LOAEL) was identified. Calculation of a reference dose was completed using standard methods based on the LD50 as a numerator with several uncertainty and modifying factors. EBD's determination of airborne concentrations aboard submarines fall in the range of these anticipated allowable concentrations and could indicate significant chronic exposures. No adverse effects from DBNP exposures have been reported to date.
Asunto(s)
Dermatitis Irritante/etiología , Dinitrofenoles/toxicidad , Desacopladores/toxicidad , Animales , Dermatitis Irritante/diagnóstico , Dinitrofenoles/análisis , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Exposición a Riesgos Ambientales/análisis , Femenino , Dosificación Letal Mediana , Masculino , Nivel sin Efectos Adversos Observados , Conejos , Ratas , Ratas Endogámicas F344 , Ratas Sprague-Dawley , Valores de Referencia , Factores de Riesgo , Medicina Submarina , Desacopladores/análisisRESUMEN
Gasoline (CAS 86290-81-5) is one of the world's largest volume commercial products. Although numerous toxicology studies have been conducted, the potential for reproductive toxicity has not been directly assessed. Accordingly, a two-generation reproductive toxicity study in rats was conducted to provide base data for hazard assessment and risk characterization. The test material, vapor recovery unit gasoline (68514-15-8), is the volatile fraction of formulated gasoline and the material with which humans are most likely to come in contact. The study was of standard design. Exposures were by inhalation at target concentrations of 5000, 10 000, and 20 000 mg/m(3). The highest exposure concentration was approximately 50% of the lower explosive limit and several orders of magnitude above anticipated exposure during refueling. There were no treatment-related clinical or systemic effects in the parental animals, and no microscopic changes other than hyaline droplet nephropathy in the kidneys of the male rats. None of the reproductive parameters were affected, and there were no deleterious effects on offspring survival and growth. The potential for endocrine modulation was also assessed by analysis of sperm count and quality as well as time to onset of developmental landmarks. No toxicologically important differences were found. Therefore, the NOAEL for reproductive toxicity in this study was > or =20 000 mg/m(3). The only systemic effects, in the kidneys of the male rats, were consistent with an alpha-2 u-globulin-mediated process. This is a male rat-specific effect and not relevant to human health risk assessment.
Asunto(s)
Contaminantes Ocupacionales del Aire/toxicidad , Gasolina/toxicidad , Efectos Tardíos de la Exposición Prenatal , Administración por Inhalación , Animales , Animales Recién Nacidos/crecimiento & desarrollo , Cámaras de Exposición Atmosférica , Peso Corporal/efectos de los fármacos , Estro/efectos de los fármacos , Femenino , Gónadas/efectos de los fármacos , Gónadas/patología , Crecimiento/efectos de los fármacos , Humanos , Masculino , Nivel sin Efectos Adversos Observados , Tamaño de los Órganos/efectos de los fármacos , Embarazo , Ratas , Ratas Sprague-Dawley , Sistema Respiratorio/efectos de los fármacos , Sistema Respiratorio/patología , Espermatozoides/efectos de los fármacos , Espermatozoides/patologíaRESUMEN
Neurotoxicity of tricresyl phosphates (TCPs) and jet engine oil (JEO) containing TCPs were evaluated in studies conducted in both rat and hen. Results for currently produced samples ("conventional" and "low-toxicity") were compared with published findings on older samples to identify compositional changes and relate those changes to neurotoxic potential. Finally, a human risk assessment for exposure by oral ingestion of currently produced TCPs in JEO at 3% (JEO + 3%) was conducted. TCPs and certain other triaryl phosphates administered as single doses inhibited brain neuropathy target esterase (B-NTE; neurotoxic esterase) in the rat and the hen (hen 3.25 times as sensitive), and both species were deemed acceptable for initial screening purposes. Neither rat nor hen was sensitive enough to detect statistically significant inhibition of B-NTE after single doses of IEO + 3% "conventional" TCP. Subacute administration of 2 g/kg/d of JEO + 3% "conventional" TCP to the hen produced B-NTE inhibition (32%), which did not result in organophosphorus-induced delayed neurotoxicity (OPIDN). Subchronic administration of JEO + 3% TCP but not JEO + 1% TCP at 2 g/kg/d produced OPIDN. Thus, the threshold for OPIDN was between 20 and 60 mg "conventional" TCP/kg/d in JEO for 10 wk. The current "conventional" TCPs used in JEO and new "low-toxicity" TCPs now used in some JEO are synthesized from phenolic mixtures having reduced levels of ortho-cresol and ortho-xylenols resulting in TCPs of very high content of meta- and para-substituted phenyl moieties; this change in composition results in lower toxicity. The "conventional" TCPs still retain enough inhibitory activity to produce OPIDN, largely because of the presence of ortho-xylyl moieties; the "low-toxicity" TCPs are largely devoid of ortho substituents and have extremely low potential to cause OPIDN. The TCPs produced in the 1940s and 1950s were more than 400 times as toxic as the "low-toxicity" TCPs produced today. Analysis of the doses required to produce OPIDN in a subchronic hen study suggests that the minimum toxic dose of "conventional" TCP for producing OPIDN in a 70-kg person would be 280 mg/d, and for JEO containing 3% TCP, 9.4 g/d. Food products could be inadvertently contaminated with neat "conventional" TCP but it is unlikely that food such as cooking oil would be contaminated with enough JEO + 3% TCP to cause toxicity. Further, at the dosage required for neurotoxicity, it would be virtually impossible for a person to receive enough JEO + 3% TCP in the normal workplace (or in an aircraft) to cause such toxicity. There is no record of a JEO formulated with the modern "conventional" TCP causing human neurotoxicity.
Asunto(s)
Encéfalo/efectos de los fármacos , Hidrolasas de Éster Carboxílico/antagonistas & inhibidores , Colinesterasas/efectos de los fármacos , Aceites Combustibles/toxicidad , Neurotoxinas/toxicidad , Tritolilfosfatos/toxicidad , Administración Oral , Animales , Ataxia/inducido químicamente , Encéfalo/enzimología , Hidrolasas de Éster Carboxílico/efectos de los fármacos , Pollos , Colinesterasas/sangre , Femenino , Aceites Combustibles/análisis , Cromatografía de Gases y Espectrometría de Masas , Humanos , Masculino , Neurotoxinas/administración & dosificación , Neurotoxinas/química , Exposición Profesional/efectos adversos , Ratas , Ratas Long-Evans , Tritolilfosfatos/administración & dosificación , Tritolilfosfatos/químicaRESUMEN
Coal, which contains significant amounts of water, can be ground and dried to produce an efficient fuel for electric power plants; however, spontaneous combustion can occur in the dried coal. Liquid petroleum hydrocarbons inhibit this combustion, but not all petroleum streams are effective. No. 6 fuel oil, a readily available and inexpensive stream, provides an effective coating, but the carcinogenic potential of coal particles treated with No. 6 fuel oil, which contains polynuclear aromatic hydrocarbons (PNAs), was undefined. As part of the assessment process, a series of studies was conducted to compare this treated coal with similar particles (petroleum coke) that had been tested by chronic inhalation in monkeys and rats. The amounts of PNAs in petroleum coke and treated coal were compared in extraction studies; the treated coal had only two-thirds of the organics extractable with benzene compared with coke and only 7% as much of the 3-7 ring PNAs, the likely tumorigenic compounds. In addition, the analytical profile of 3-7 ring PNAs was of lower molecular weights in the coal treated with fuel oil. The mutagenicity of extracts from treated coal was much less than with petroleum coke and markedly less than that of No. 6 fuel oil itself. The percutaneous absorption of 3H-benzo(a)pyrene from both particles and from their benzene extracts, as measured in vitro, was approximately eight times greater with petroleum coke than with treated coal. Based on these preliminary results, there is no evidence suggesting that the treated coal would pose any greater carcinogenic risk than petroleum coke.
Asunto(s)
Carcinógenos/análisis , Carbón Mineral/toxicidad , Aceites Combustibles/toxicidad , Hidrocarburos Policíclicos Aromáticos/análisis , Absorción Cutánea , Aerosoles , Animales , Disponibilidad Biológica , Pruebas de Carcinogenicidad/métodos , Carbón Mineral/análisis , Coque/análisis , Coque/toxicidad , Femenino , Aceites Combustibles/análisis , Técnicas In Vitro , Tamaño de la Partícula , Ratas , Ratas Sprague-DawleyRESUMEN
A structure-activity relationship (SAR) of the in vitro percutaneous absorption of polycyclic aromatic hydrocarbons (PAH) is described. The data set consisted of 60 three to seven ring PAH. Over 50 numeric descriptors were generated from the modeled molecular structures. Computer aided methods were used to evaluate descriptors and develop linear expressions relating the percent of dermally applied PAH dose absorbed through skin (PADA) to PAH structure. Three regression models with one and two variables were developed. The log octanol/water partition coefficient (log P) was the most important variable in determining percutaneous absorption. An inverse relationship between log P and the skin penetration properties of the PAH was observed. Nearly 40 of 60 PAH tested had PADA-values within a factor of two of benzo[a]pyrene (BaP); well over 50 of 60 had PADA-values within a factor of three. The results lend support to the use of isotopically labeled BaP as a surrogate for measuring the dermal flux (in vivo and in vitro) and estimating the dermal bioavailability of PAH from complex mineral oil and coal-tar derived mixtures.
Asunto(s)
Exposición a Riesgos Ambientales , Hidrocarburos Policíclicos Aromáticos/farmacocinética , Absorción , Administración Cutánea , Disponibilidad Biológica , Humanos , Modelos Logísticos , Relación Estructura-ActividadRESUMEN
Two crude oils, differing in viscosity (V) and nitrogen (N) and sulfur (S) content, were evaluated for pre- and postnatal developmental toxicity. In Crude I (low V, low N, low S) studies, the material was applied neat to the clipped backs of pregnant rats at dose levels of 0, 125, 500, 1000 (postnatal only), and 2000 (prenatal only) mg/kg. In Crude II (high V, high N, moderate S) studies, the oil was applied similarly but at dose levels of 0, 30, 125, and 500 mg/kg. Rats were exposed to the crude oils on gestation days (GD) 0-19; application sites were not covered. "Prenatal" rats were killed on GD 20. "Postnatal" rats were allowed to deliver naturally; surviving dams and litters were killed 3-4 wk postpartum. Both crude oils produced maternal and developmental toxicity. Adverse fetal effects included increased in utero death, decreased body weight, and reduced ossification of skeletal elements. Parturition was delayed in Crude II dams at 500 mg/kg. The 4-d viability index was decreased in all Crude II-exposed groups. Pup body weights were decreased by each oil, but at the high dose only. Prenatal effects are probably related to polynuclear aromatic compounds (PAC) found in petroleum. The cause(s) of delayed parturition and postnatal toxicity have not been determined.
Asunto(s)
Desarrollo Embrionario y Fetal/efectos de los fármacos , Feto/efectos de los fármacos , Petróleo/toxicidad , Anomalías Inducidas por Medicamentos , Administración Cutánea , Animales , Animales Recién Nacidos , Peso Corporal/efectos de los fármacos , Desarrollo Óseo/efectos de los fármacos , Dermatitis Irritante/patología , Femenino , Edad Gestacional , Masculino , Embarazo , Ratas , Piel/efectos de los fármacos , Piel/patología , Tasa de SupervivenciaRESUMEN
An in vitro system was utilized to measure DNA adduct-forming ability of petroleum oils and oil coal tar mixtures to define correlations between DNA adduct levels and their mutagenic potencies. The system consisted of reaction of dimethyl sulfoxide extracts of oils with calf thymus DNA in the presence of Aroclor-induced hamster liver microsomes for 30 min. Following DNA extraction, DNA adducts were measured by the nuclease P1-enhanced postlabeling assay coupled with two-dimensional polyethyleneimine (PEI)-cellulose TLC. Thin layer plates showed putative aromatic DNA adducts, with levels ranging from 60 to 1400 adducts per 10(9) DNA nucleotides. TLC mobilities suggested adducts to be aromatic compounds containing 4 or more rings. A good correlation (coefficient of correlation = 0.91) was observed between DNA adduct levels and Salmonella mutagenicity for 19 oils. All 19 samples tested produced DNA adducts. To expedite the TLC procedure, adducts were resolved by one-dimensional TLC and the radioactivity measured using a mechanical scanner. Results were comparable to those obtained by two-dimensional TLC and quantification after scraping. Our data show that the in vitro incubation system coupled with the postlabeling adduct assay is a useful screening method to identify mutagenic and potentially carcinogenic oils.
Asunto(s)
Alquitrán/toxicidad , Aductos de ADN/metabolismo , Mutágenos/toxicidad , Petróleo/toxicidad , Animales , Arocloros/farmacología , Autorradiografía , Carcinógenos/metabolismo , Carcinógenos/toxicidad , Bovinos , Cromatografía en Capa Delgada , Alquitrán/metabolismo , Cricetinae , ADN/metabolismo , Técnicas In Vitro , Microsomas Hepáticos/metabolismo , Pruebas de Mutagenicidad/métodos , Petróleo/metabolismo , Radioisótopos de Fósforo/metabolismo , Salmonella/metabolismo , Endonucleasas Específicas del ADN y ARN con un Solo Filamento/metabolismo , Timo/químicaRESUMEN
Two crude oils, differing in viscosity (V) and nitrogen (N) and sulfur (S) content, were evaluated for systemic toxicity. In the Crude I (low V, low N, low S) study, the material was applied to the clipped backs of rats at dose levels of 0, 30, 125, and 500 mg/kg. In the Crude II (high V, high N, moderate S) study, the oil was applied similarly at the same dose levels. The crude oils were applied for 13 wk, 5 d/wk. Exposure sites were not occluded. Mean body weight gain (wk 1-14) was significantly reduced in male rats exposed to Crude II; body weight gain of all other animals was not adversely affected by treatment. An increase in absolute (A) and relative (R) liver weights and a decrease in A and R thymus weights were observed in male and female rats exposed to Crude II at 500 mg/kg; only liver weights (A and R) were adversely affected in male and female rats exposed to Crude I. In general, there was no consistent pattern of toxicity for serum chemistry endpoints; however, more parameters were adversely affected in Crude II-exposed female rats than in the other exposed groups. A consistent pattern of toxicity for hematology endpoints was observed among male rats exposed to Crude I and male and female rats exposed to Crude II. Parameters affected included: Crudes I and II, red blood cell count, hemoglobin, and hematocrit; Crude II, platelet count. Microscopic evaluation of tissues revealed the following treatment-related findings: Crude I, treated skin, thymus, and thyroid; Crude II, bone marrow, treated skin, thymus, and thyroid. The LOEL (lowest observable effect level) for skin irritation and systemic toxicity (based on marginal effects on the thyroid) for both crude oils was 30 mg/kg; effects were more numerous and more pronounced in animals exposed to Crude II. Systemic effects are probably related to concentrations of polycyclic aromatic compounds (PAC) found in crude oil.
Asunto(s)
Hígado/efectos de los fármacos , Petróleo/toxicidad , Administración Cutánea , Animales , Peso Corporal/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Hígado/patología , Masculino , Tamaño de los Órganos/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Piel/efectos de los fármacos , Piel/patología , Relación Estructura-ActividadRESUMEN
Syntower bottoms (STB) was evaluated for subchronic and developmental toxicity. In the subchronic study, undiluted STB was applied on the shaved backs of male and female rats at dose levels of 0, 8, 30, 125, and 500 mg/kg for 13 weeks, 5 days per week. Exposure sites were not covered. In the developmental toxicity study, STB was similarly applied, but to pregnant rats at dose levels of 0, 8, 30, and 125 mg/kg on Gestation Days 0-19. In addition, 4 mg/kg was dosed as 8 mg/kg every other day, starting on Gestation Day 0, and 500 mg/kg was dosed on Gestation Days 10-12. Evidence of toxicity observed in the subchronic study included death, decreased body weights, aberrant serum chemistry and hematology values, altered organ weights, and histopathologic changes in a variety of organs. A no observed adverse effect level for systemic toxicity could not be established. Evidence of maternal toxicity was observed at all exposure levels in the development study. Regardless of the length of the exposure period, STB was toxic to the developing conceptus. Evidence of developmental toxicity observed included increased resorptions with a concomitant decrease in litter size and reduced fetal body weights. Cleft palate was observed in fetuses exposed in utero to STB during Gestation Days 10-12 at 500 mg/kg. No evidence of teratogenicity was observed when the exposure period was throughout gestation. Ossification delays were observed in fetuses exposed in utero to STB at doses in excess of 4 mg/kg. A no observed adverse effect level for maternal and developmental toxicity could not be established.
Asunto(s)
Industrias , Petróleo , Compuestos Policíclicos/toxicidad , Anomalías Inducidas por Medicamentos/patología , Administración Tópica , Animales , Recuento de Células Sanguíneas/efectos de los fármacos , Peso Corporal/efectos de los fármacos , Desarrollo Óseo/efectos de los fármacos , Desarrollo Embrionario y Fetal/efectos de los fármacos , Enzimas/sangre , Femenino , Lípidos/sangre , Masculino , Compuestos Policíclicos/administración & dosificación , Compuestos Policíclicos/química , Embarazo , Ratas , Ratas Sprague-Dawley , Reproducción/efectos de los fármacos , Aumento de Peso/efectos de los fármacosRESUMEN
The Zymbal gland, a sebaceous tissue associated with the ear duct of certain rodent species, is a principal target for carcinogenesis by benzene. To investigate the mechanism of induction of tumors in the rat Zymbal gland, we have developed a procedure for primary culture of epithelial cells from Zymbal gland explants so that cytogenetic analysis can be performed on this target tissue following an in vivo exposure to benzene. Cytogenetic analysis performed 45 hr after in vivo oral dosing with benzene revealed chromosome damage that occurred as a result of acute, subchronic, and chronic dosing. This damage, expressed as a dose-related increase in the frequency of micronucleated cells, was observed in Sprague-Dawley female rats over a range of benzene doses from 12.5 to 250 mg/kg/day, and in male Fischer 344 rats at doses ranging from 1 to 200 mg/kg/day. These results are consistent with the known clastogenicity of benzene in mouse bone marrow, which is also a target tissue. This study is the first report of a genotoxic effect of benzene in the rat Zymbal gland and shows that micronucleus formation may be used as a correlate for carcinogenesis induced by benzene in this target tissue.
Asunto(s)
Benceno/toxicidad , Glándulas Sebáceas/efectos de los fármacos , Animales , Benceno/administración & dosificación , Médula Ósea/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Masculino , Ratones , Pruebas de Micronúcleos , Mutágenos/administración & dosificación , Mutágenos/toxicidad , Ratas , Ratas Endogámicas F344 , Ratas Sprague-Dawley , Glándulas Sebáceas/ultraestructuraRESUMEN
Clarified slurry oil (CSO, CAS number 64741-62-4), a refinery stream produced by processing crude oil, is a developmental toxicant when administered dermally throughout gestation to pregnant rats. The manifestations of developmental toxicity observed included embryolethality and growth retardation; evidence of teratogenicity was limited, and not conclusive. The present study was undertaken to further explore the teratogenic potential of CSO. In an attempt to limit embryolethality and thereby promote detection of terata, CSO was administered once daily for a limited period of gestation [gestation days (GD) 9-12], via dermal application, to pregnant Sprague-Dawley rats at doses of 0, 10, 100, and 1000 mg/kg. All animals were sacrificed on GD 20. Detailed examination of the dams was performed. Due to the screening nature of this investigation, fetal evaluations were limited to body weight measurements, external examinations, and evaluation of select visceral endpoints. In the dams exposed to CSO, significant decreases in body weight [absolute and gain (GD 9-13, GD 0-20)] and in the amount of food consumed were observed at 100 and 1000 mg/kg. Additional evidence of maternal toxicity observed at 1000 mg/kg included decreased absolute and relative thymus weights, increased absolute and relative liver weights, and aberrant serum chemistry. Ingestion of the test material was evident at the high dose. Developmental toxicity was observed at 1000 mg/kg and included increased embryolethality, decreased body weight, and anomalous development (cleft palate, brachydactyly, edema). Although a low incidence of abnormal fetal development was observed at 100 mg/kg, it was not conclusive that the alterations were due to CSO exposure. It is likely that three to seven-ring polycyclic aromatic compounds present in CSO were responsible for the toxic effects observed.
Asunto(s)
Carbazoles/toxicidad , Desarrollo Embrionario y Fetal/efectos de los fármacos , Petróleo/toxicidad , Compuestos Policíclicos/toxicidad , Preñez/efectos de los fármacos , Administración Cutánea , Animales , Análisis Químico de la Sangre , Peso Corporal/efectos de los fármacos , Carbazoles/administración & dosificación , Relación Dosis-Respuesta a Droga , Ingestión de Alimentos/efectos de los fármacos , Femenino , Feto/anomalías , Feto/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/patología , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/patología , Masculino , Tamaño de los Órganos/efectos de los fármacos , Compuestos Policíclicos/administración & dosificación , Embarazo , Ratas , Ratas Sprague-Dawley , Reproducción/efectos de los fármacos , Bazo/efectos de los fármacos , Bazo/patología , Timo/efectos de los fármacos , Timo/patologíaRESUMEN
Methyl tertiary-butyl ether (MTBE), which is added to gasoline as an octane enhancer and to reduce automotive emissions, has been evaluated in numerous toxicological tests, including those for genotoxicity. MTBE did not show any mutagenic potential in the Ames bacterial assay or any clastogenicity in cytogenetic tests. However, it has been shown to be mutagenic in an in vitro gene mutation assay using mouse lymphoma cells when tested in the presence, but not in the absence, of a rat liver-derived metabolic activation system (S-9). In the present study, MTBE was tested to determine if formaldehyde, in the presence of the S-9, was responsible for the observed mutagenicity. A modification of the mouse lymphoma assay was employed which permits determination of whether a suspect material is mutagenic because it contains or is metabolized to formaldehyde. In the modified assay, the enzyme formaldehyde dehydrogenase (FDH) and its co-factor, NAD+ are added in large excess during the exposure period so that any formaldehyde produced in the system is rapidly converted to formic acid which is not genotoxic. An MTBE dose-responsive increase in the frequency of mutants and in cytotoxicity occurred without FDH present, and this effect was greatly reduced in the presence of FDH NAD+. The findings clearly demonstrate that formaldehyde derived from MTBE is responsible for mutagenicity of MTBE in the activated mouse lymphoma assay. Furthermore, the results suggest that the lack of mutagenicity/clastogenicity seen with MTBE in other in vitro assays might have resulted from inadequacies in the test systems employed for those assays.
Asunto(s)
Formaldehído/metabolismo , Formaldehído/toxicidad , Éteres Metílicos/metabolismo , Éteres Metílicos/toxicidad , Mutágenos/metabolismo , Mutágenos/toxicidad , Solventes/metabolismo , Solventes/toxicidad , Animales , Gasolina , Técnicas In Vitro , Leucemia L5178 , Ratones , Microsomas Hepáticos/metabolismo , Pruebas de Mutagenicidad/métodos , RatasRESUMEN
Clarified slurry oil (CSO), syntower bottoms (STB), and distillate aromatic extract (DAE) are refinery streams produced by processing crude oil. Each of these refinery streams is rich in both hydrocarbons having carbon numbers of C20 or greater and polycyclic aromatic compounds. Available data indicate that some refinery streams are developmentally toxic (manifested primarily as increased embryolethality and growth retardation) by the dermal route of exposure. However, there is no conclusive evidence for their being teratogenic. The present studies were designed to further explore the suspected teratogenic potency of refinery streams while at the same time limiting embryolethality. To profile teratogenic effects as a function of gestation day, pregnant rats received a single oral dose (2000 mg/kg) of CSO, STB, or DAE on one of gestation days (GD) 11-14; DAE and STB were also administered on GD 15. To profile effects as a dose response function, rats received a single oral dose of CSO, DAE, or STB on GD 12 at 125, 500, and 2000 mg/kg. Control animals were similarly treated but were administered tap water. On GD 20, dams were necropsied and the fetuses evaluated for normal development. In general, evidence of maternal toxicity (i.e., decreased body weight gain, decreased thymus weight) was observed at doses greater than or equal to 500 mg/kg. For each refinery stream tested, the incidence of resorption was greatest on GD 11. A common pattern of fetal malformations was observed for all of the refinery streams tested and included cleft palate, diaphragmatic hernia, and paw and tail defects. The incidence and type of malformation observed were influenced by the gestation day of exposure. The incidences of external and skeletal malformations were greatest on GD 11 and 12 for fetuses exposed to CSO; on GD 13 and 14, the incidence of malformation was comparable for CSO- and STB-exposed fetuses. The incidence of visceral anomalies was greatest on GD 11-13 for fetuses exposed to CSO and STB; on Gestation D 14, the incidence was comparable for each of the refinery streams tested. In general, the ability to produce adverse effects on development was greatest for CSO and least for DAE. Effects produced by STB were comparable to or less severe than those observed for CSO.
Asunto(s)
Anomalías Inducidas por Medicamentos , Huesos/anomalías , Carbazoles/toxicidad , Desarrollo Embrionario y Fetal/efectos de los fármacos , Petróleo/toxicidad , Compuestos Policíclicos/toxicidad , Administración Oral , Animales , Peso Corporal/efectos de los fármacos , Huesos/efectos de los fármacos , Carbazoles/administración & dosificación , Relación Dosis-Respuesta a Droga , Pérdida del Embrión , Femenino , Masculino , Tamaño de los Órganos/efectos de los fármacos , Compuestos Policíclicos/administración & dosificación , Embarazo , Ratas , Reproducción/efectos de los fármacosRESUMEN
Distillate aromatic extract (DAE) was evaluated for subchronic and developmental toxicity. In the subchronic study, undiluted DAE was applied on the shaved backs of male and female rats at dose levels of 0, 30, 125, 500, and 1250 mg/kg for 13 weeks, 5 days per week. Exposure sites were not covered. In the developmental toxicity study, DAE was similarly applied, but to pregnant rats at dose levels of 0, 8, 30, and 125 mg/kg on Gestation Days 0-19, 500 mg/kg on Gestation Days 0-16, and 1000 mg/kg on Gestation Days 10-12. Evidence of toxicity observed in the subchronic study included death, decreased body weights, aberrant serum chemistry and hematology values, altered organ weights, and histopathologic changes in a variety of organs. Regardless of the length of the exposure period, DAE was toxic to the developing conceptus. Evidence of developmental toxicity observed included increased resorptions and reduced fetal body weights. Cleft palate and ossification delays were observed in fetuses exposed in utero to DAE on Gestation Days 10-12, but not when exposure spanned all (Gestation Days 0-19) or most (Gestation Days 0-16) of gestation.
Asunto(s)
Hígado/efectos de los fármacos , Petróleo/toxicidad , Teratógenos/toxicidad , Animales , Desarrollo Óseo/efectos de los fármacos , Médula Ósea/efectos de los fármacos , Fisura del Paladar/inducido químicamente , Relación Dosis-Respuesta a Droga , Desarrollo Embrionario y Fetal/efectos de los fármacos , Femenino , Pruebas Hematológicas , Masculino , Embarazo , Ratas , Absorción Cutánea , Timo/efectos de los fármacosRESUMEN
The potential of benzene to produce DNA adducts in B6C3F1 mice was investigated by the nuclease P1-enhanced 32P-postlabeling assay. The mice were daily treated i.p. with 500 mg/kg benzene in olive oil for 4 days, and the liver, mammary gland, and bone marrow were collected 24 h after the last treatment. Thin-layer chromatograms obtained with treated-tissue DNA specimens were qualitatively identical to those from corresponding olive oil-treated (control) tissue DNA. Quantitative evaluations revealed that there was no treatment-related increase in radioactivity on the chromatograms at or near the locations where the major in vitro adducts of phenol, hydroquinone and benzoquinone migrated. Benzene treatment, however, resulted in a decrease in the levels of certain endogenous adducts, the biological significance of which is unknown. Our results indicate that benzene treatment does not produce detectable levels of aromatic DNA adducts in mouse tissues.
Asunto(s)
Benceno/farmacología , Aductos de ADN , Animales , Médula Ósea/química , Femenino , Hígado/química , Glándulas Mamarias Animales/química , RatonesRESUMEN
Refinery streams are complex mixtures, but of a relatively few homologous series of hydrocarbons (paraffins, olefins, naphthenics, and aromatics). Studies were performed to determine if systemic and developmental toxicity were related to the presence and levels of certain classes of refinery stream components. We have performed systemic toxicology studies in the rat on 13 refinery streams: Clarified Slurry Oil, Coker Light Gas Oil, Distillate Aromatic Extract, Heavy Atmospheric Gas Oil, Heavy Coker Gas Oil (from three refineries), Heavy Vacuum Gas Oil, Light Catalytically Cracked Naphtha, Light Cycle Oil, Syntower Bottoms, Vacuum Tower Overhead, and Visbreaker Gas Oil. Rats were exposed via repeated dermal administration (daily) at several dose levels. Developmental toxicology studies were performed on these same streams with the following exceptions: only two Heavy Coker Gas Oils were tested and Visbreaker Gas Oil was not tested. End points for systemic toxicity (13-week) studies included skin irritation, body and organ weights, hematology, and serum chemistry; for developmental toxicity studies some of these same end points (excluding hematology) were considered, but they also included resorption and fetal body weight. In general, toxicity was correlated with concentrations of polycyclic aromatic compounds (PAC) composed of 3, 4, 5, 6, and/or 7 rings (decreased thymus weight, increased liver weight, aberrant hematology and serum chemistry, increased incidence of resorption, decreased fetal body weight), PAC containing nonbasic nitrogen heteroatoms (increased mortality, decreased body weight, decreased thymus weight, increased liver weight, decreased hemoglobin content and hematocrit level, decreased fetal body weight), and/or PAC containing sulfur heteroatoms (decreased red blood cell and platelet counts, increased sorbitol dehydrogenase.)(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Carbazoles/toxicidad , Desarrollo Embrionario y Fetal/efectos de los fármacos , Petróleo/clasificación , Petróleo/toxicidad , Compuestos Policíclicos/toxicidad , Animales , Carbazoles/análisis , Femenino , Hígado/efectos de los fármacos , Masculino , Pruebas de Mutagenicidad , Tamaño de los Órganos/efectos de los fármacos , Petróleo/análisis , Compuestos Policíclicos/análisis , Ratas , Ratas Sprague-DawleyRESUMEN
An in vitro eye irritation test battery (IVEye) composed of the EYTEX and Modified Agarose Diffusion Method (MADM) assays was evaluated for use as a predictive, economical screen and/or adjunct for the Draize eye test. EYTEX mimics corneal opacification using a synthetic matrix of proteins that is intended to produce measurable opacity on exposure to chemical irritants in proportion to their ocular irritation potential. MADM is a cytotoxicity-based assay consisting of NCTC clone 929 mouse fibroblasts overlayed with 1% agarose in culture medium. Potential eye irritation is measured macroscopically as the area of decolorization (neutral red release) around the area of chemical application and microscopically as the percentage of cell lysis resulting from chemical application. Of the 70 materials tested in the IVEye for which Draize eye test data also exist, the battery correctly identified 38 materials as non-irritants and 30 as irritants, with two false positives and no false negatives. Non-parametric analysis of the data show the battery to have a sensitivity of 100%, a specificity of 95% and a predictive value of 94%. The irritation class correlation (equivalence; irritation ranking) between EYTEX alone and the Draize data was 85%. These data support the use of IVEye as an accurate, reproducible and cost-effective in vitro method for identifying the eye irritation potential of petroleum products.
RESUMEN
89 petroleum products were tested in the EYTEX in vitro assay and the results were compared with data from Draize tests on the same materials. All 26 chemicals assessed as irritant in the Draize test were classified as irritant by the EYTEX assay; 61 of the 63 non-irritants were correctly classified by the in vitro assay. The correlation between the results from the EYTEX system and the Draize data was 89%, and the predictive value of the EYTEX system (percentage of EYTEX irritants that are true irritants) was 93%. When the same chemicals were tested in another laboratory, the coefficient of determination for the comparison of the EYTEX scores from the two laboratories was R(2) = 0.86. The results demonstrate the reliability and interpretability of the EYTEX assay carried out in our laboratory and therefore the viability, if not the validity, of this in vitro method as a screening test for the Draize assay.
RESUMEN
The carcinogenicity of benzene has been considered to be in part mediated by its chemically reactive metabolic product benzoquinone (BQ), which is formed from the intermediary metabolites phenol and hydroquinone (HQ). We have evaluated the DNA-binding capability of these chemicals in vitro and in vivo by postlabeling. Treatment of rat Zymbal glands in culture with phenol and HQ or direct reaction of BQ with DNA produced DNA adducts, which were detectable by the nuclease P1-enhanced 32P-postlabeling assay as 5'-32P-labeled 3',5'-bisphosphate products. The enhancement of sensitivity in this assay is based on the previous finding that nuclease P1 hydrolyzes the phosphate attached to the 3' side of normal nucleotides but not the corresponding phosphate of most aromatic/bulky adducted nucleotides. Also based on this hydrolytic property of nuclease P1, we developed an additional sensitive procedure that permitted the detection of DNA lesions as 5'-32P-labeled products of dinucleotides, pXpN, or of nucleoside monophosphates, pX, where X and N indicate an adducted nucleoside and a normal nucleoside respectively. In the latter assay, adducted DNA was first digested with nuclease P1 and acid phosphatase to yield XpN and N. The latter were then 32P-labeled to yield [5'-32P] pXpN or 32P-labeled and treated with venom phosphodiesterase to obtain [5'-32P]pX. After optimization of enzymatic conditions, the modified nuclease P1 assay yielded adduct recoveries similar to those obtained by the bisphosphate assay for in vitro phenol-, HQ- and BQ-DNA adducts. Neither of the nuclease P1-enhanced postlabeling procedures showed exposure-specific adducts in vivo in the bone marrow, Zymbal gland, liver and spleen of female Sprague-Dawley rats at 24 h after the last of four single, daily p.o. doses of 75 mg/kg phenol or 150 mg/kg phenol/HQ (1:1). Our results show that phenol, HQ and BQ produce adducts in vitro, but corresponding adducts are not detected in vivo with phenol and phenol/HQ, even when measured by the standard and modified nuclease P1 postlabeling methods capable of detecting 1 adduct in 10(9-10) DNA bases.