RESUMO
The operating environment of the service personnel during the Persian Gulf War involved psychological, biological, and chemical elements including exposure to pesticides such as the insect repellent DEET (N,N-diethyl-m-toluamide) and the insecticide chlorpyrifos (O,O-diethyl O-3,5,6-trichloropyridinyl phosphorothioate) and to pyridostigmine bromide (PB,3-dimethylaminocarbonyloxy-N-methylpyridinium bromide) that was administered as a prophylactic agent against possible nerve gas attack. The present study was designed to determine the toxicity produced by individual or coexposure of hens 5 days/week for 2 months to 5 mg PB/kg/day in water, by gavage; 500 mg DEET/kg/day, neat, sc; and 10 mg chlorpyrifos kg/day in corn oil, sc. Coexposure to various binary treatments produced greater neurotoxicity than that caused by individual exposures and was characterized by severe neurologic deficit and neuropathological alterations. Also, neurotoxicity was further enhanced following concurrent administration of the three chemicals. Severe inhibition of plasma butyrylcholinesterase (BuChE) activity was produced in hens treated with PB (activity 17% of control) compared to those treated with chlorpyrifos (activity 51% of control) or DEET (activity 83% of control). BuChE inhibition was further increased in binary and tertiary treatment groups compared to individual treatment groups. In contrast, a significant inhibition of brain acetylcholinesterase (AChE) was produced in hens administered chlorpyrifos alone (activity 67% of control), while those given chlorpyrifos in combination with other compounds exhibited a significant inhibition of brain AChE activity ranging from 43 to 76%. Brain neurotoxicity target esterase (NTE) was not inhibited in any of the individual treatment groups or PB/DEET, but was significantly inhibited and had activity expressed as a percentage of control in groups administered combined chlorpyrifos with PB of 73% or DEET of 74% and in the tertiary treatment group of 71%. We hypothesize that test compounds may compete for xenobiotic metabolizing enzymes in the liver and blood and may also compromise the integrity of the blood-brain barrier, leading to an increase in their "effective concentrations" in the nervous system to levels equivalent to the toxic doses of individual compounds. This is consistent with the present observation of increases in (1) the inhibition of brain AChE and NTE, (2) the extent of neurologic dysfunction, and (3) the severity and frequency of neuropathologic lesions in the combined treatment groups compared to those administered individual compounds.
Assuntos
Galinhas/fisiologia , Clorpirifos/toxicidade , Inibidores da Colinesterase/toxicidade , DEET/toxicidade , Repelentes de Insetos/toxicidade , Doenças do Sistema Nervoso/induzido quimicamente , Brometo de Piridostigmina/toxicidade , Animais , Encéfalo/enzimologia , Encéfalo/patologia , Colinesterases/sangue , Colinesterases/metabolismo , Interações Medicamentosas , Discinesia Induzida por Medicamentos/patologia , Doenças do Sistema Nervoso/enzimologia , Doenças do Sistema Nervoso/patologia , Medula Espinal/enzimologia , Medula Espinal/patologiaRESUMO
Of the three-quarters of a million service personnel involved in the Persian Gulf War, approximately 30,000 have complained of neurological symptoms of unknown etiology. One contributing factor to the emergence of such symptoms may be the simultaneous exposure to multiple agents used to protect the health of service personnel, in particular, the anti-nerve agent pyridostigmine bromide (PB; 3-dimethylaminocarbonyloxy-N-methylpyridinium bromide), the insect repellent DEET (N,N-diethyl-m-toluamide), and the insecticide permethrin (3-(2,2-dichloro-ethenyl)-2,2-dimethylcyclopropanecarboxylic acid (3-phenoxyphenyl)methyl ester). This study investigated neurotoxicity produced in hens by individual or simultaneous exposure to these agents (5 d/wk for 2 months to 5 mg/kg/d PB in water, po; 500 mg/kg/d DEET, neat, sc; and 500 mg/kg/d permethrin in corn oil, sc). At these dosages, exposure to single compounds resulted in minimal toxicity. Combinations of two agents produced greater neurotoxicity than that caused by individual agents. Neurotoxicity was further enhanced following concurrent administration of all three agents. We hypothesize that competition for liver and plasma esterases by these compounds leads to their decreased breakdown and increased transport of the parent compound to nervous tissues. Thus, carbamylation of peripheral esterases by PB reduces the hydrolysis of DEET and permethrin and increases their availability to the nervous system. In effect, PB "pumps" more DEET and permethrin into the central nervous system. Consistent with this hypothesis, hens exposed to the combination of the three agents exhibited neuropathological lesions with several characteristics similar to those previously reported in studies of near-lethal doses of DEET and permethrin. If this hypothesis is correct, then blood and liver esterases play an important "buffering" role in protecting against neurotoxicity in the population at large. It also suggests that individuals with low plasma esterase activity may be predisposed to neurologic deficits produced by exposure to certain chemical mixtures.
Assuntos
Inibidores da Colinesterase/toxicidade , DEET/toxicidade , Repelentes de Insetos/toxicidade , Inseticidas/toxicidade , Sistema Nervoso/efeitos dos fármacos , Piretrinas/toxicidade , Brometo de Piridostigmina/toxicidade , Administração Oral , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/enzimologia , Guerra Química , Galinhas , Inibidores da Colinesterase/administração & dosagem , Colinesterases/sangue , Colinesterases/metabolismo , DEET/administração & dosagem , Sinergismo Farmacológico , Feminino , Humanos , Injeções Subcutâneas , Repelentes de Insetos/administração & dosagem , Inseticidas/administração & dosagem , Sistema Nervoso/patologia , Sistema Nervoso/fisiopatologia , Exposição Ocupacional , Permetrina , Piretrinas/administração & dosagem , Brometo de Piridostigmina/administração & dosagem , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/patologia , Medula Espinal/efeitos dos fármacos , Medula Espinal/patologia , Distribuição TecidualRESUMO
A single oral dose of [2,3-14C]acrylamide (50 mg/kg) was administered in water to adult white leghorn hens. Seven groups of three hens were euthanized between 2 and 120 hr after administration. Within 12 hr, the hens excreted 70% of the administered dose, and more than 99% within 48 hr. Blood, plasma, liver, and muscle contained the greatest percentage of administered dose at 4 hr after dosing. Less than 0.02% of the administered dose appeared in brain at any time. Radiolabel accumulated in the eggs, with 0.52% of the administered dose accumulated within 5 days. Binding of radiolabel to erythrocytes was minimal. Elimination of radiolabel from all tissues was biphasic. Terminal elimination half-lives for 14C were longer than 10 days, at which time less than 0.2% of the administered dose remains in the tissues. Distribution half-lives for 14C were longest for whole blood and shortest for kidney. Radioactivity in the blood and plasma reached a peak at between 4 and 12 hr. Most of this radioactivity was identified as acrylamide, which disappeared biexponentially with terminal elimination half-lives longer than 10 days. Distribution half-lives for acrylamide were longest in brain and shortest in whole blood. These results show that orally administered acrylamide is poorly absorbed and rapidly eliminated from hens and accumulates in their eggs in a nonextractable form.
Assuntos
Acrilamidas/farmacocinética , Acrilamida , Acrilamidas/administração & dosagem , Acrilamidas/sangue , Acrilamidas/toxicidade , Tecido Adiposo/metabolismo , Animais , Química Encefálica , Galinhas , Ovos/análise , Feminino , Meia-Vida , Rim/metabolismo , Fígado/metabolismo , Músculo Esquelético/metabolismo , Ligação ProteicaRESUMO
This study was carried out to determine the action of glycidamide (2,3-epoxy-1-propanamide), a neurotoxic metabolite of acrylamide, on Ca2+/calmodulin (CaM)-dependent protein kinase phosphorylation of cytoskeletal proteins. Acrylamide has been shown to increase Ca2+/CaM-dependent phosphorylation of neurofilament (NF) triplet proteins and autophosphorylation of Ca2+/CaM-dependent protein kinase II (CaM kinase II; EC 2.7.1.37). A daily intraperitoneal dose of 0.7 mmol/kg b.wt. of glycidamide or deionized water was administered to male Sprague-Dawley rats. Animals were sacrificed when signs of severe neurotoxicity became apparent at 13-16 days of treatment. Axonal floatation was used to isolate neurofilaments (NFs) and endogenous kinases from brains and spinal cords of treated and control animals. Samples isolated from brain and spinal cord of glycidamide-treated animals showed increased in vitro Ca2+/CaM-dependent phosphorylation of endogenous and exogenous NF proteins and increased autophosphorylation of CaM kinase II when compared with controls. CaM binding to the alpha, beta, and beta' subunits of CaM kinase II and antibody binding to the alpha-subunit of CaM kinase II in brain supernatant isolates was increased as a result of glycidamide treatment. These results suggest that increased Ca2+/CaM-dependent phosphorylation of cytoskeletal proteins may be involved in the pathogenesis of glycidamide-induced neurotoxicity.
Assuntos
Química Encefálica , Encéfalo/metabolismo , Proteínas Quinases Dependentes de Cálcio-Calmodulina/farmacologia , Compostos de Epóxi/farmacologia , Proteínas de Neurofilamentos/metabolismo , Medula Espinal/química , Acrilamida , Acrilamidas/farmacologia , Animais , Encéfalo/enzimologia , Imunoensaio , Técnicas In Vitro , Masculino , Proteínas de Neurofilamentos/efeitos dos fármacos , Neurotoxinas/farmacologia , Fosforilação , Ratos , Ratos Sprague-Dawley , Medula Espinal/enzimologiaRESUMO
2,5-Hexanedione (2,5-HD) induces central-peripheral axonpathy characterized by the accumulation of 10-nm neurofilaments proximal to the nodes of Ranvier and a Wallerian-type degeneration. It has been postulated that neurofilament crosslinking may be involved in the production of this axonopathy. A potential initiating event in this neurotoxic process may be the direct binding of 2,5-HD to neurofilament and microtubule proteins. In this study, the in vitro binding of [14C]2,5-HD to neurofilament and microtubule proteins was examined. Neurofilament proteins isolated from rat spinal cord or microtubule proteins isolated from rat brain were incubated in the presence of 2,5-HD at concentrations ranging from 25 to 500 mM. Quantitative analysis of sodium dodecyl sulfate (SDS) polyacrylamide gels revealed a dose- and time-dependent binding of 2,5-HD to both neurofilament proteins and microtubule proteins. Expressed as pmol 2,5-HD bound per microgram protein, the observed relative binding was MAP2 > NF160 > NF200 > > NF68 > tubulin. These data demonstrate the direct binding of 2,5-HD to cytoskeletal proteins including both neurofilaments and microtubules.
Assuntos
Proteínas do Citoesqueleto/metabolismo , Hexanonas/metabolismo , Proteínas dos Microtúbulos/metabolismo , Proteínas de Neurofilamentos/metabolismo , Neurônios/metabolismo , Animais , Radioisótopos de Carbono , Immunoblotting , Masculino , Oxirredução , Pirróis/metabolismo , Ensaio Radioligante , Ratos , Ratos Sprague-DawleyRESUMO
Male Sprague-Dawley rats were administered a daily i.p. dose of 0.70 mmol/kg body weight of acrylamide, propionamide (a non-neurotoxic structural analog of acrylamide) or deionized water. Animals were sacrificed when signs of severe neurotoxicity were apparent. Neurofilaments (NFs) and endogenous kinase were isolated from the brain and spinal cord by axonal floatation. Increased in vitro Ca2+/calmodulin-dependent phosphorylation of endogenous and exogenous NF proteins and autophosphorylation of Ca2+/calmodulin protein kinase II (CaM kinase II, EC 2-7-1-37) were observed in samples from both brain and spinal cord of acrylamide-treated animals compared with controls. There was no significant difference between samples isolated from propionamide-treated animals and controls. Increased calmodulin binding to brain supernatant CaM kinase II was also observed as a result of acrylamide treatment. There was no significant difference observed in the amount of antibody binding to the alpha-subunit of brain supernatant CaM kinase II between treated or control animals. These results suggest that increased CaM kinase II-dependent phosphorylation of cytoskeletal proteins may be involved in the mechanisms of acrylamide-induced neurotoxicity.
Assuntos
Acrilamidas/farmacologia , Encéfalo/metabolismo , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Proteínas de Neurofilamentos/metabolismo , Medula Espinal/metabolismo , Acrilamida , Acrilamidas/toxicidade , Amidas/farmacologia , Animais , Encéfalo/efeitos dos fármacos , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina , Calmodulina/metabolismo , Masculino , Doenças do Sistema Nervoso/induzido quimicamente , Fosforilação , Ratos , Ratos Sprague-Dawley , Medula Espinal/efeitos dos fármacosRESUMO
The Ca2+/calmodulin-dependent phosphorylation of neuronal cytoskeletal proteins was studied in brain supernatants prepared from rats exposed via inhalation to 600 to 800 ppm carbon disulfide (CS2) for 14 days. Exposure to CS2 resulted in increased phosphorylation of endogenous MAP-2 and exogenously added neurofilament triplet proteins. There also was an observed increase in the autophosphorylation of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II). Slight increases in the binding of a monoclonal antibody to the alpha subunit of CaM kinase II were seen, while large increases in the binding of [125I]calmodulin to the alpha subunit of CaM kinase II also were observed. The finding of large increases in the autophosphorylation and calmodulin-binding to CaM kinase II with only slight increases in the amount of antibody-binding suggests that CS2 exposure results in increased Ca2+/calmodulin-dependent phosphorylation of proteins by inducing an increase in kinase activity.
Assuntos
Encéfalo/efeitos dos fármacos , Proteínas Quinases Dependentes de Cálcio-Calmodulina/efeitos dos fármacos , Dissulfeto de Carbono/farmacologia , Proteínas do Citoesqueleto/metabolismo , Administração por Inalação , Animais , Encéfalo/enzimologia , Encéfalo/metabolismo , Imunoensaio , Masculino , Proteínas de Neurofilamentos/metabolismo , Fosforilação , Ensaio Radioligante , Ratos , Ratos Sprague-DawleyRESUMO
Inhibition of lysyl oxidase (protein-lysine 6-oxidase, EC 1.4.3.13) decreases the rate of collagen and elastin cross-link formation and produces osteolathyrism in animals. Organic nitriles, including beta-aminopropionitrile (BAPN), have been shown to irreversibly inhibit lysyl oxidase in vitro. Both BAPN and 3,3'-iminodipropionitrile (IDPN) have been shown to produce osteolathyric changes when administered to animals. To date compounds that have been reported to inhibit this enzyme possess a primary amine functional group. In this study a series of primary and substituted aminopropionitriles was studied for their ability to inhibit lysyl oxidase activity both in vitro and in vivo. Our results show that of the compounds tested, BAPN was the most potent inhibitor of the enzyme. Reversible inhibition of lysyl oxidase in vitro was found with two secondary aminonitriles, IDPN and monomethylaminopropionitrile (MMAPN). There was no inhibition of enzyme activity associated with the tertiary compound 3,3'-dimethylaminopropionitrile (DMAPN) or propionitrile, a compound lacking an amine functional group. IDPN was found to produce a slight irreversible inhibition of the enzyme both in vitro and in vivo. Pretreatment of rats with pargyline, an inhibitor of monoamine oxidase, was found to increase the inhibitory potential of BAPN (p < or = .1). Pargyline pretreatment did not alter the inhibitory potential for any of the other aminonitriles tested. These results suggest that the presence of a primary amino functional group is not a strict requirement for inhibition of lysyl oxidase. In addition, reversible and irreversible mechanisms of inhibition may be involved in the production of osteolathyric changes associated with IDPN exposure.
Assuntos
Aminopropionitrilo/farmacologia , Nitrilas/farmacologia , Proteína-Lisina 6-Oxidase/antagonistas & inibidores , Animais , Aorta/efeitos dos fármacos , Aorta/enzimologia , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/enzimologia , Embrião de Galinha , Técnicas In Vitro , Masculino , Ratos , Ratos Sprague-Dawley , TrítioRESUMO
Oxidation of aminopropionitriles was measured in vitro with both rat liver mitochondria and bovine plasma monoamine oxidase (MAO). The nonneurotoxic aminonitrile beta-aminopropionitrile (BAPN) was oxidized at a significantly higher rate (p less than .05) than either of the neurotoxic aminonitriles tested; 3,3'-iminodipropionitrile (IDPN) and 3,3'-dimethylaminopropionitrile (DMAPN). DMAPN was a poor substrate for both mitochondrial and plasma MAO. None of the aminonitriles tested were found to inhibit MAO activity in rat brain or liver in vivo. Inhibition of MAO activity with pargyline in vivo did not affect the pattern of IDPN- or DMAPN-induced toxicity. These results suggest that monoamine oxidase is not involved in aminonitrile-induced neurotoxicity.