RESUMEN
Although discovered more than a century ago, piscine rodlet cells (RCs) remain somewhat of a mystery to scientists in terms of their origin and function. Initially described as parasites, and later as potential secretory cells, the prevailing theory is that RCs are leucocyte-like cells that possess pathogen defence capabilities. The current case report involves a novel type of neoplasm discovered in the livers of two adult female white suckers (Catostomus commersonii) that were collected as part of a survey of fish from the St. Mary's River Area of Concern, in which sediment contaminated by polyaromatic hydrocarbons has been associated historically with a high prevalence of liver neoplasms in white suckers. The two tumours in this study were investigated by light microscopy, histochemical staining, immunohistochemical labelling for S100 protein and transmission electron microscopy. The evidence from these investigations suggests that these neoplasms may be derived from de-differentiated RCs or RC precursors. The unanticipated existence of these solid mesenchymal-like tumours may prompt a reassessment of the current dogma regarding the physiological function of RCs.
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Cipriniformes , Enfermedades de los Peces/patología , Neoplasias Hepáticas/veterinaria , Animales , FemeninoRESUMEN
A 13-week-old male intact Poodle mix dog developed an acute onset of vestibular ataxia, tetraparesis, and vomiting. The patient presented ambulatory, tetraparetic, and ataxic with a head tilt to the left and a disconjugate nystagmus (rotary nystagmus with fast phase to the right in right eye and vertical nystagmus in left eye). There were absent postural reactions in the left pelvic and left thoracic limbs and decreased right-sided postural reactions. Magnetic resonance imaging demonstrated an intra-axial mass within the left midcaudal medulla oblongata. On gross dissection, there was a left-sided neoplasm in the medulla oblongata with surrounding hemorrhage. The histologic findings indicated that the mass was a pleomorphic xanthoastrocytoma. This tumor, an uncommon variant of an astrocytoma most often seen in children and young adult humans, has yet to be described in dogs.
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Astrocitoma/veterinaria , Neoplasias Encefálicas/veterinaria , Enfermedades Vestibulares/veterinaria , Animales , Astrocitoma/patología , Neoplasias Encefálicas/patología , Perros , Imagen por Resonancia Magnética/veterinaria , Masculino , Bulbo Raquídeo/patología , Enfermedades Vestibulares/patologíaRESUMEN
BACKGROUND: In humans, central neurocytomas are rare and typically benign intracranial tumors found within the lateral ventricles, although extraventricular variants have been reported. Intracranial central neurocytomas have not been previously recognized in domestic animals. OBJECTIVES: To describe the clinicopathologic features of canine intracranial central neurocytomas. ANIMALS: Two dogs with spontaneous intracranial and intraventricular neoplasms. RESULTS: Both dogs experienced seizures, rapid neurological deterioration, and death from tumor-associated complications within 5 days of the onset of clinical signs, and had neoplastic masses within the lateral ventricles. A brain MRI was performed in 1 dog, which revealed a T1-isointense, heterogeneously T2 and FLAIR hyperintense, and markedly and heterogeneously contrast-enhancing mass lesions within both lateral ventricles. Histologically, the neoplasms resembled oligodendrogliomas. The diagnosis of central neurocytoma was supported by documenting expression of multiple neuronal markers, including neuron-specific enolase, synaptophysin, neural-cell adhesion molecule, and neuronal nuclear antigen within the tumors, and ultrastructural evidence of neuronal differentiation of neoplastic cells. CONCLUSIONS AND CLINICAL IMPORTANCE: Central neurocytoma should be a differential diagnosis for dogs with intraventricular brain masses. Morphologic differentiation of central neurocytoma from other intraventricular neoplasms, such as ependymoma or oligdendroglioma, can be difficult, and definitive diagnosis often requires immunohistochemical or ultrastructural confirmation of the neural origin of the neoplasm.
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Neoplasias Encefálicas/veterinaria , Enfermedades de los Perros/patología , Neurocitoma/veterinaria , Animales , Western Blotting/veterinaria , Neoplasias Encefálicas/patología , Neoplasias Encefálicas/ultraestructura , Perros , Resultado Fatal , Inmunohistoquímica/veterinaria , Masculino , Microscopía Electrónica de Transmisión/veterinaria , Neurocitoma/patología , Neurocitoma/ultraestructuraRESUMEN
Cytoplasmic vacuolation of sensory neurons has been reported to occur within the dorsal root ganglia in studies investigating various neuropathic conditions including the effects of neurotoxic chemicals. In this study, we investigated this lesion in adult (98-119 days old) male Long-Evans rats, after multiple exposures to two organophosphates (tri-ortho-tolyl phosphate [TOTP] and chlorpyrifos) and the modifying effects of concurrent corticosterone. Tri-ortho-tolyl phosphate was administered by gavage (75, 150, or 300 mg/kg) every other day between days 14 and 28 and between days 49 and 63, chlorpyrifos (60 mg/kg) was administered subcutaneously on days 7 and 42, and corticosterone was provided in the drinking water throughout the study at a concentration of 400 microg/mL. Although relatively uncommon, there was an increase in frequency of cytoplasmic vacuoles seen in treatment groups having multiple exposures to TOTP. They were characterized as peripherally located, single-limiting membrane-bound structures in the neuronal perikarya. There was no associated cell death, even when vacuoles were large. This is the initial report of an association of this change following exposure to neurotoxic organophosphates.
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Cloropirifos/toxicidad , Citoplasma/efectos de los fármacos , Ganglios Sensoriales/efectos de los fármacos , Células Receptoras Sensoriales/efectos de los fármacos , Tritolilfosfatos/toxicidad , Vacuolas/efectos de los fármacos , Animales , Corticosterona/farmacología , Interacciones Farmacológicas , Ganglios Sensoriales/citología , Histocitoquímica , Masculino , Microscopía Electrónica , Ratas , Ratas Long-Evans , Células Receptoras Sensoriales/citología , Pruebas de Toxicidad Crónica/métodosRESUMEN
Circulating uranium rapidly enters the brain and may cause adverse effects on the nervous system that are potentially modulated by stress. In this study, the neurological effects of a single intramuscular injection of 0, 0.1, 0.3, or 1 mg uranium/kg (as uranyl acetate, UA) in rats were examined in the presence and absence of stress. Treatment with UA produced time and dose-dependent increases in serum and regional brain uranium levels. While serum levels returned to control levels by day 30, brain levels remained elevated. Application of stress did not affect the distribution or retention of uranium. Exposure to 1 mg U/kg significantly decreased ambulatory activity, weight gain, forelimb grip strength and transiently impaired working memory. Effects on grip strength and memory were prevented by application of stress prior to uranium exposure. Striatal dopamine content was reduced by 30% 3 days after treatment with 1mg/kg (59+/-6 nmol/mg tissue versus 41+/-5 nmol/mg tissue), but levels returned to control 7 days after uranium exposure. The effect on dopamine was ameliorated by prior application of stress. Exposure to UA did not alter 3,4 dihydroxyphenylacetic acid (DOPAC) levels or numbers of D2 receptors in the striatum. No effect of uranium or stress was observed on levels of GABA, serotonin, norepinephrine, or glutathione (GSH) in the striatum, hippocampus, cerebellum, or cortex. These results indicate that single intramuscular exposures to uranium produce sustained elevation of brain uranium levels and at doses above 0.3 mg/kg can have adverse neurological effects. Application of stress prior to uranium administration modulates neurological effects, but the mechanism is not due to effects on uranium distribution. Uranium exposure also produced renal toxicity which must be considered to accurately assess the effects of uranium on neurological function.
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Conducta Animal/efectos de los fármacos , Encéfalo/efectos de los fármacos , Síndromes de Neurotoxicidad/etiología , Compuestos Organometálicos/toxicidad , Estrés Psicológico/complicaciones , Animales , Peso Corporal/efectos de los fármacos , Encéfalo/metabolismo , Encéfalo/fisiopatología , Química Encefálica/efectos de los fármacos , Corticosterona/sangre , Dopamina/metabolismo , Relación Dosis-Respuesta a Droga , Glutatión/metabolismo , Inyecciones Intramusculares , Enfermedades Renales/inducido químicamente , Locomoción/efectos de los fármacos , Masculino , Memoria/efectos de los fármacos , Fuerza Muscular/efectos de los fármacos , Síndromes de Neurotoxicidad/complicaciones , Síndromes de Neurotoxicidad/metabolismo , Síndromes de Neurotoxicidad/fisiopatología , Neurotransmisores/metabolismo , Compuestos Organometálicos/administración & dosificación , Compuestos Organometálicos/farmacocinética , Ratas , Ratas Sprague-Dawley , Estrés Psicológico/metabolismo , Estrés Psicológico/fisiopatología , Factores de Tiempo , Distribución TisularRESUMEN
Exposure to OP compounds that inhibit neurotoxic esterase (NTE) induces a delayed neuropathy (OPIDN) characterized by Wallerian-like degeneration of long axons in certain animals, including humans. Pope et al. (Toxicol. Lett. 75:111-117, 1995) found that neurite outgrowth occurred following the addition of spinal cord extracts from chickens with active OPIDN to neuroblastoma cells, suggesting growth factor expression during the neuropathy. We hypothesized that, shortly after exposure to a neuropathic OP compound, the central nervous system (CNS) attempts to recover from the toxic insult through upregulation of the neurotrophins nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) in susceptible regions of the nervous system. We hypothesized that such upregulation is transient and cannot be sustained. To test this hypothesis, we exposed 10-week-old chickens to a neuropathic OP compound (PSP, 2.5 mg/kg), a non-neuropathic OP compound (paraoxon, 0.10 mg/kg), and vehicle (DMSO, 0.5 ml/kg) intramuscularly. By day 8, all PSP-treated birds demonstrated clinical signs of OPIDN. We sacrificed chickens by pentobarbital overdose at 4, 8, 24, and 48 hours, and 5 and 10 days post-exposure and confirmed NTE inhibition in birds treated with PSP 4 and 24 hours earlier. Enzyme-linked immunosorbant assays indicated that NGF, BDNF, and NT-3 are found in chicken lumbar spinal cord after exposure to a neuropathic OP compound. However, exposure to the neuropathic OP compound, PSP, did not preferentially elevate levels of NGF, BDNF, and NTE compared to the non-neuropathic OP compound, paraoxon. This suggests that these neurotrophins alone do not contribute to a sustained regenerative effort in the CNS.
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Factores de Crecimiento Nervioso/metabolismo , Enfermedades del Sistema Nervioso/inducido químicamente , Enfermedades del Sistema Nervioso/metabolismo , Compuestos Organofosforados/toxicidad , Degeneración Walleriana/inducido químicamente , Degeneración Walleriana/metabolismo , Animales , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Pollos , Inhibidores de la Colinesterasa/toxicidad , Factor de Crecimiento Nervioso/metabolismo , Regeneración Nerviosa , Neurotrofina 3/metabolismo , Paraoxon/toxicidadRESUMEN
Recent studies in vivo and in vitro suggested that mitochondrial dysfunction follows exposure to organophosphorus (OP) esters. As mitochondrial ATP production is important for cellular integrity, ATP production in the presence of OP neurotoxicants was examined in a human neuronal cell line (SH-SY5Y neuroblastoma cells) and primary dorsal root ganglia (DRG) cells isolated from chick embryos and subsequently cultured to achieve maturation with axons. These cell culture systems were chosen to evaluate toxic effects on the mitochondrial respiratory chain associated with exposure to OP compounds that do and do not cause OP-induced delayed neuropathy (OPIDN), a disorder preceded by inhibition of neurotoxic esterase (NTE). Concentration- and time-response studies were done in neuroblastoma cells exposed to phenyl saligenin phosphate (PSP) and mipafox, both compounds that readily induce delayed neuropathy in hens, or paraoxon, which does not. Phenylmethylsulfonyl fluoride (PMSF) was included as a non-neuropathic inhibitor of NTE. Purified neuronal cultures from 9 day-old chick embryo DRG were treated for 12 h with 1 microM PSP, mipafox, or paraoxon. In situ evaluation of ATP production measured by bioluminescence assay demonstrated decreased ATP concentrations both in neuroblastoma cells and chick DRG neurons treated with PSP. Mipafox decreased ATP production in DRG but not in SH-SY5Y cells. This low energy state was present at several levels of the mitochondrial respiratory chain, including Complexes I, II, III, and IV, although Complex I was the most severely affected. Paraoxon and PMSF were not effective at all complexes, and, when effective, required higher concentrations than needed for PSP. Results suggest that mitochondria are an important early target for OP compounds, with exposure resulting in depletion of ATP production. The targeting of neuronal, rather than Schwann cell mitochondria in DRG following exposure to PSP and mipafox was verified by loss of the mitochondrial-specific dye, tetramethylrhodamine, in these cells. No such loss was seen in paraoxon exposed neurons isolated from DRG or in Schwann cells treated with any of the test compounds.
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Adenosina Trifosfato/metabolismo , Inhibidores de la Colinesterasa/toxicidad , Isoflurofato/análogos & derivados , Mitocondrias/efectos de los fármacos , Neuronas/efectos de los fármacos , Compuestos Organofosforados/toxicidad , Animales , Células Cultivadas , Embrión de Pollo , Relación Dosis-Respuesta a Droga , Ganglios Espinales/citología , Humanos , Microscopía Confocal/métodos , Mitocondrias/metabolismo , Complejos Multienzimáticos/metabolismo , Neuroblastoma , Neuronas/metabolismo , Paraoxon , Fluoruro de Fenilmetilsulfonilo , Células de Schwann/efectos de los fármacos , Factores de TiempoRESUMEN
An important method of assessing experimental neurotoxic injury is the pathologic examination of the nervous system. Methods for fixation, sampling, and preparation of peripheral nervous system tissues for critical pathological neurotoxicology studies are presented. Fixation of tissue is carried out using either perfusion-fixation of laboratory animals or immersion-fixation of dissected nerve segments. Dissection of the peripheral nervous system (from perfusion-fixed animals) is done to allow for multilevel sampling. Focus is on use of epoxy resin embedding tissue sections for optimal light microscopic resolution. Protocols for processing, sectioning, and staining for light and transmission electron microscopy are provided. A protocol for teasing and microscopic study of individual myelinated fibers is provided.
Asunto(s)
Neurotoxinas/toxicidad , Sistema Nervioso Periférico/ultraestructura , Animales , Microscopía Electrónica de Transmisión , Sistema Nervioso Periférico/efectos de los fármacos , RatasRESUMEN
An evaluation of potential adverse human health effects of disinfection byproducts requires study of both cancer and noncancer endpoints; however, no studies have evaluated the neurotoxic potential of a common haloacetic acid, dibromoacetic acid (DBA). This study characterized the neurotoxicity of DBA during 6-month exposure in the drinking water of rats. Adolescent male and female Fischer 344 rats were administered DBA at 0, 0.2, 0.6, and 1.5 g/l. On a mg/kg/day basis, the consumed dosages decreased greatly over the exposure period, with average intakes of 0, 20, 72, and 161 mg/kg/day. Weight gain was depressed in the high-concentration group, and concentration-related diarrhea and hair loss were observed early in exposure. Testing with a functional observational battery and motor activity took place before dosing and at 1, 2, 4, and 6 months. DBA produced concentration-related neuromuscular toxicity (mid and high concentrations) characterized by limb weakness, mild gait abnormalities, and hypotonia, as well as sensorimotor depression (all concentrations), with decreased responses to a tail-pinch and click. Other signs of toxicity at the highest concentration included decreased activity and chest clasping. Neurotoxicity was evident as early as one month, but did not progress with continued exposure. The major neuropathological finding was degeneration of spinal cord nerve fibers (mid and high concentrations). Cellular vacuolization in spinal cord gray matter (mostly) and in white matter (occasionally) tracts was also observed. No treatment-related changes were seen in brain, eyes, peripheral nerves, or peripheral ganglia. The lowest-observable effect level for neurobehavioral changes was 20 mg/kg/day (produced by 0.2 g/l, lowest concentration tested), whereas this dosage was a no-effect level for neuropathological changes. These studies suggest that neurotoxicity should be considered in the overall hazard evaluation of haloacetic acids.
Asunto(s)
Acetatos/efectos adversos , Administración Oral , Síndromes de Neurotoxicidad/etiología , Abastecimiento de Agua/análisis , Acetatos/administración & dosificación , Acetatos/farmacocinética , Alopecia/inducido químicamente , Animales , Conducta Animal/efectos de los fármacos , Temperatura Corporal/efectos de los fármacos , Desinfectantes/efectos adversos , Desinfectantes/química , Desinfectantes/farmacocinética , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Evaluación Preclínica de Medicamentos/métodos , Femenino , Masculino , Actividad Motora/efectos de los fármacos , Degeneración Nerviosa/diagnóstico , Degeneración Nerviosa/patología , Síndromes de Neurotoxicidad/diagnóstico , Ratas , Ratas Endogámicas F344 , Factores Sexuales , Nervios Espinales/efectos de los fármacos , Nervios Espinales/patología , Nervios Espinales/ultraestructura , Factores de Tiempo , Contaminación Química del Agua/efectos adversos , Contaminación Química del Agua/análisis , Aumento de Peso/efectos de los fármacosRESUMEN
An animal (rat) model of chronic stress (corticosterone in the drinking water) was used to study the interaction of stress and the organophosphorus (OP) neurotoxicants chlorpyrifos (60 mg/kg subcutaneously in a single dose) and tri-ortho-tolyl phosphate (TOTP, at 75, 150, or 300 mg/kg given 7 times orally in a 2-wk period). Adult male Long-Evans rats were provided with corticosterone in drinking water (400 microg/ml, w/v) for a total of 28 d, which led to significantly decreased weight and decreased cellularity of the thymus and spleen. Seven days after initiation of corticosterone treatment, half of the rats were given chlorpyrifos, and an additional 7 d later the 2-wk, 7-dose treatment of TOTP was initiated. During the 28-d test period, behavior of rats was evaluated using a functional observational battery (FOB), motor activity, and passive avoidance. Reductions in body weight, grip strength, and ambulatory movements occurred as a result of corticosterone treatment. Decreased body weight and grip strength were also elicited by TOTP, and the interactions of corticosterone and TOTP enhanced the effects on body weight and grip strength. Blood cholinesterase levels were obtained during the 28-d study period and found useful for monitoring OP exposure. At the end of the 28-d testing period, rats were sacrificed and activities of cholinesterase, neurotoxic esterase (neuropathy target esterase), and/or carboxylesterase were evaluated in blood, liver, and/or brain regions (basal forebrain, caudate putamen, cerebral cortex, hippocampus). All these esterases in brain were inhibited in a dose-related manner by TOTP, with some enhancement in rats drinking corticosterone-containing water. In addition, choline acetyltransferase, glial acidic fibrillary protein (GFAP), glutathione peroxidase, and superoxide dismutase were evaluated in one or more of the brain regions already identified. Choline acetyltransferase, glutathione peroxidase, and superoxide dismutase activities were unaffected by treatments. However, GFAP was elevated above control levels in the cerebral cortex of rats by all treatments (corticosterone, chlorpyrifos, TOTP). Neuropathological examination revealed early stages of dose-related increased distal myelinated fiber axonal degeneration seen in the medullary fasciculus gracilis at only the highest dose of TOTP (300 mg/kg).
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Antiinflamatorios/farmacología , Cloropirifos/farmacología , Corticosterona/farmacología , Insecticidas/farmacología , Tritolilfosfatos/toxicidad , Animales , Axones/efectos de los fármacos , Axones/patología , Peso Corporal/efectos de los fármacos , Encéfalo/efectos de los fármacos , Encéfalo/enzimología , Corteza Cerebral/metabolismo , Colinesterasas/sangre , Relación Dosis-Respuesta a Droga , Ingestión de Líquidos , Esquema de Medicación , Esterasas/metabolismo , Proteína Ácida Fibrilar de la Glía/metabolismo , Masculino , Modelos Animales , Degeneración Nerviosa , Fibras Nerviosas Mielínicas/efectos de los fármacos , Fibras Nerviosas Mielínicas/patología , Ratas , Ratas Long-EvansRESUMEN
To assess the relationship of nerve conduction and adenosine triphosphate (ATP) status in organophosphorus-induced delayed neuropathy (OPIDN), we evaluated both in adult hen peripheral nerves following exposure to a single 2.5 mg/kg dose of phenyl saligenin phosphate (PSP). ATP concentrations were determined at days 2, 4, 7, and 14 post-dosing, from five segments (n = 5 per group) representing the entire length of the sciatic-tibial and medial plantar nerve. Initial effects of PSP dosing were seen in the most distal segment at day 2, when a transient ATP concentration increase (388 +/- 79 pmol/ml/mg versus control value of 215 +/- 23, P < 0.05) was noted. Subsequently, ATP concentration in this distal segment returned to normal. In the most proximal nerve segment, ATP concentrations were decreased on day 7, and further decreased on day 14 post-dosing (P < 0.05). Changes in ATP concentration and nerve conduction velocity begin at post-dosing day 2, and were found prior to development of clinical neuropathy and axonopathic lesions. These results suggest that alterations in sciatic-tibial and medial plantar nerve conduction associated with sciatic-tibial and medial plantar nerve ATP concentration are early events in the development of OPIDN.
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Adenosina Trifosfato/metabolismo , Pollos/metabolismo , Pie/inervación , Insecticidas/toxicidad , Conducción Nerviosa/efectos de los fármacos , Síndromes de Neurotoxicidad/metabolismo , Compuestos Organofosforados/toxicidad , Nervio Ciático/efectos de los fármacos , Nervio Ciático/metabolismo , Nervio Tibial/efectos de los fármacos , Nervio Tibial/metabolismo , Potenciales de Acción/efectos de los fármacos , Animales , Conducta Animal/efectos de los fármacos , Electrofisiología , Femenino , Factores de TiempoRESUMEN
Neuropathologic assessment of chemically induced developmental alterations in the nervous system for regulatory purposes is a multifactorial, complex process. This calls for careful qualitative and quantitative morphologic study of numerous brains at several developmental stages in rats. Quantitative evaluation may include such basic methods as determination of brain weight and dimensions as well as the progressively more complex approaches of linear, areal, or stereologic measurement of brain sections. Histologic evaluation employs routine stains (such as hematoxylin and eosin), which can be complemented by a variety of special and immunohistochemical procedures. These brain studies are augmented by morphologic assessment of selected peripheral nervous system structures. Studies of this nature require a high level of technical skill as well as special training on the part of the pathologist. The pathologist should have knowledge of normal microscopic neuroanatomy/neuronal circuitry and an understanding of basic principles of developmental neurobiology, such as familiarity with the patterns of physiologic or programmed cell de
Asunto(s)
Conducta Animal/efectos de los fármacos , Encéfalo/efectos de los fármacos , Encéfalo/crecimiento & desarrollo , Sistema Nervioso Periférico/efectos de los fármacos , Sistema Nervioso Periférico/crecimiento & desarrollo , Xenobióticos/efectos adversos , Animales , Biometría , Humanos , Ratas , Medición de Riesgo , Manejo de Especímenes , Fijación del TejidoRESUMEN
Organophosphorus (OP) compounds have been shown to be cytotoxic to SH-SY5Y human neuroblastoma cell cultures. The mechanisms involved in OP compound-induced cell death (apoptosis versus necrosis) were assessed morphologically by looking at nuclear fragmentation and budding using the fluorescent stain Hoechst 33342 (10 microgram/ml). Hoechst staining revealed significant paraoxon (1 mM), parathion (1 mM), phenyl saligenin phosphate (PSP, 10 and 100 microM), tri-ortho-tolyl phosphate (TOTP, 100 microM and 1 mM), and triphenyl phosphite (TPPi, 1 mM) induced time-dependent increases in traditional apoptosis (p < 0.05). In many cells, PSP and TOTP (1 mM) also induced nuclear condensation with little fragmentation or budding. Pretreatment with cyclosporin A (500 nM, 30 h) decreased apoptosis following 1 mM parathion and TOTP exposures. Apoptotic nuclear changes were verified by DNA gel electrophoresis. Activation of caspase-3, a cysteine aspartate protease, was also monitored. OP compounds induced significant time-dependent increases in caspase-3 activation following paraoxon (1 mM), parathion (100 microM, 1 mM), PSP (10 microM, 100 microM, 1 mM), TOTP (100 microM, 1 mM), and TPPi (1 mM) exposure (p < 0.05). Pretreatment with cyclosporin A (500 nM, 30 h) significantly decreased caspase-3 activation during extended incubations with paraoxon, parathion, and TPPi (p < 0.05). In addition, pretreatment with the caspase-3 inhibitor Ac-DEVD-CHO and the caspase-8 inhibitor Ac-IETD-CHO (25 microM, 8 h) significantly decreased caspase-3 activation following exposure to 1 mM PSP and parathion (p < 0.05). Pretreatment with the serine protease inhibitor phenylmethyl sulfonyl fluoride (PMSF; 1 mM, 8 h) also significantly decreased caspase activation following 1 mM PSP and TOTP exposures (p < 0.05). Alteration of OP compound-induced nuclear fragmentation or caspase-3 activation by pretreatment with cyclosporin A, Ac-IETD-CHO, or PMSF suggested that OP compound-induced cytotoxicity may be modulated through multiple sites, including mitochondrial permeability pores, receptor-mediated caspase pathways, or serine proteases.
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Apoptosis/efectos de los fármacos , Inhibidores de la Colinesterasa/toxicidad , Neuronas/efectos de los fármacos , Compuestos Organofosforados/toxicidad , Caspasa 3 , Caspasas/metabolismo , Fragmentación del ADN , Electroforesis en Gel de Agar , Activación Enzimática/efectos de los fármacos , Humanos , Insecticidas/toxicidad , Microscopía Electrónica , Microscopía Fluorescente , Necrosis , Neuroblastoma , Neuronas/citología , Neuronas/patología , Células Tumorales CultivadasRESUMEN
OBJECTIVE: To evaluate the clinical, cerebrospinal fluid (CSF), and histopathologic effects of epidural ketorolac. STUDY DESIGN: Blinded, randomized, placebo controlled study. ANIMALS: Twenty-two adult mixed breed dogs with 16 treatment and 6 control dogs, weighing 14.4 to 29.8 kg. METHODS: Dogs were anesthetized and epidural catheters were placed at the lumbosacral space. Catheter placement was evaluated fluoroscopically. Ketorolac (0.4 mg/kg) or placebo (5% ethanol) was administered epidurally over a 52-hour period, with 5 injections given at 12-hour intervals. At 1, 2, 4, or 8 hours after the first and last injection of ketorolac, dogs were anesthetized and CSF was obtained. Control dogs had CSF sampled 1 hour after the first and last ethanol injection. Neurologic function and pain responses were evaluated before and during the study. Selected dogs were then killed and necropsies performed. RESULTS: None of the dogs exhibited any clinical or neurologic abnormalities during the study. No statistical difference was noted in pain response or CSF analysis between treatment and control dogs. Gross necropsy revealed gastrointestinal ulceration of varying degrees in all treatment dogs. Histopathologic analysis of the spinal cord and meninges revealed minimal focal leptomeningeal phlebitis in 2 of 8 treatment dogs and minor subdural inflammation in 1 control dog. No changes to the neural structures were noted in any dogs. CONCLUSIONS: Epidural administration of ketorolac did not cause clinical signs, alteration in CSF values, or pathologic changes to the spinal cord when used for short duration. Gastrointestinal ulceration was common when ketorolac was administered epidurally at 0.4 mg/kg every 12 hours for 5 treatments. CLINICAL RELEVANCE: This study documented the neurologic safety of epidural ketorolac in dogs before an efficacy trial can be performed. Gastrointestinal ulceration may limit use to short duration or a single injection.
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Anestesia Epidural/veterinaria , Antiinflamatorios no Esteroideos/farmacología , Líquido Cefalorraquídeo/efectos de los fármacos , Perros/fisiología , Ketorolaco/farmacología , Meninges/efectos de los fármacos , Médula Espinal/efectos de los fármacos , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Recuento de Células Sanguíneas/efectos de los fármacos , Recuento de Células Sanguíneas/veterinaria , Líquido Cefalorraquídeo/química , Líquido Cefalorraquídeo/citología , Proteínas del Líquido Cefalorraquídeo/análisis , Proteínas del Líquido Cefalorraquídeo/efectos de los fármacos , Sistema Digestivo/efectos de los fármacos , Sistema Digestivo/patología , Perros/líquido cefalorraquídeo , Femenino , Ketorolaco/administración & dosificación , Masculino , Dolor/prevención & control , Dolor/veterinaria , Úlcera Péptica/inducido químicamente , Úlcera Péptica/veterinaria , Valores de Referencia , Método Simple CiegoRESUMEN
Exposure to a variety of agricultural, industrial, and pharmaceutical chemicals produces nerve damage classified as a central-peripheral distal axonopathy. Morphologically, this axonopathy is characterized by distal axon swellings and secondary degeneration. Over the past 25 years substantial research efforts have been devoted toward deciphering the molecular mechanisms of these presumed hallmark neuropathic features. However, recent studies suggest that axon swelling and degeneration are related to subchronic low-dose neurotoxicant exposure rates (i.e., mg toxicant/kg/day) and not to the development of neurophysiological deficits or behavioral toxicity. This suggests these phenomena are nonspecific and of uncertain pathophysiologic relevance. This possibility has significant implications for research investigating mechanisms of neurotoxicity, development of exposure biomarkers, design of risk assessment models, neurotoxicant classification schemes, and clinical diagnosis and treatment of toxic neuropathies. In this commentary we will review the evidence for the dose-related dependency of distal axonopathies and discuss how this concept might influence our current understanding of chemical-induced neurotoxicities.
Asunto(s)
Axones/efectos de los fármacos , Neurotoxinas/administración & dosificación , Polineuropatías/inducido químicamente , Xenobióticos/administración & dosificación , Animales , Axones/patología , Relación Dosis-Respuesta a Droga , Humanos , Neurotoxinas/toxicidad , Polineuropatías/patología , Xenobióticos/toxicidadRESUMEN
Quantitative morphometric analysis was used to characterize expression of myelinated axon swelling and atrophy in rat peripheral nerve during 2,5-hexanedione (HD) intoxication. HD was administered by gavage according to different daily dosing regiments (100, 175, 250, or 400 mg/kg/day) and four proximodistal nerve regions (5th lumbar spinal nerve, proximal and distal sciatic nerve, and tibial nerve) were examined morphometrically. Morphometric determinations were made at four behavioral endpoints (unaffected, slight, moderate, and severe toxicity) and were correlated to electrophysiologic measurements of peripheral nerve function. Results show that, for all HD dose rates, onsets of behavioral neurotoxicity and nerve dysfunction were generally related to development of abundant axon atrophy. The proximodistal manifestation of atrophy was dependent upon the dosing rate; i.e., the atrophy response produced by subacute intoxication with higher daily dosing rates (250 and 400 mg/kg/day) was restricted to distal nerve regions whereas subchronic induction with lower dosing rates (100 and 175 mg/kg/day) produced abundant fiber atrophy in all proximodistal areas. In contrast to atrophy, axonal swellings constituted an inconsistent minor morphologic response, the expression of which was dependent upon subchronic dosing rates (100-250 mg/kg/day). Subacute HD administration (400 mg/kg/day) produced significant changes in neurobehavior and nerve electrophysiologic parameters in the absence of peripheral axon swelling. Thus, conditional expression of swellings suggests they are an epiphenomenon related to low-dose induction rates. Fiber atrophy, however, was numerically dominant, correlated with nerve dysfunction, and occurred at all dosing levels. These characteristics suggest atrophy is a neurotoxicologically significant feature of gamma-diketone peripheral neuropathy.
Asunto(s)
Axones/patología , Hexanonas/toxicidad , Neurotoxinas/toxicidad , Enfermedades del Sistema Nervioso Periférico/inducido químicamente , Animales , Atrofia/patología , Axones/ultraestructura , Peso Corporal/efectos de los fármacos , Tamaño de la Célula/efectos de los fármacos , Electrofisiología , Marcha/efectos de los fármacos , Fuerza de la Mano/fisiología , Procesamiento de Imagen Asistido por Computador , Masculino , Vaina de Mielina/patología , Enfermedades del Sistema Nervioso Periférico/patología , Ratas , Ratas Sprague-Dawley , Nervio Tibial/patologíaRESUMEN
The anatomical distribution and organization of the peripheral nervous system as well as its frequent ability to reflect neurotoxic injury make it useful for the study of nerve fiber and ganglionic lesions. Contemporary neuropathologic techniques provide sections with excellent light-microscopic resolution for use in making such assessments. The histopathologist examining such peripheral nerve samples may see several patterns of neurotoxic injury. Most common are axonopathies, conditions in which axonal alterations are noted; these axonopathies often progress toward the Wallerian-like degeneration of affected fibers. These are usually more severe in distal regions of the neurite, and they affect both peripheral and central fibers. Examples of such distal axonopathies are organophosphorous ester-induced delayed neuropathy, hexacarbon neuropathy, and p-bromophenylacetylurea intoxication. These axonopathies may have varying pathologic features and sometimes have incompletely understood toxic mechanisms. In such neuropathies with fiber degeneration, peripheral nerve axons may regenerate, which can complicate pathologic interpretation of neurotoxicity. On occasion neurotoxins elicit more severe injury in proximal regions of the fiber (not included in this review). Axonal pathology is also a feature of the neuronopathies, toxic states in which the primary injuries are found in neuronal cell bodies. This is exemplified by pyridoxine neurotoxicity, where there is sublethal or lethal damage to larger cytons in the sensory ganglia, with failure of such neurons to maintain their axons. Lastly, one may encounter myelinopathies, conditions in which the toxic effect is on the myelin-forming cell or sheath. An example of this is tellurium intoxication, where demyelination noted in young animals is coincident with toxin-induced interference of cholesterol synthesis by Schwann cells. In this paper, the above-noted examples of toxic neuropathy are discussed, with emphasis on mechanistic and morphologic considerations.
Asunto(s)
Nervios Periféricos/patología , Enfermedades del Sistema Nervioso Periférico/inducido químicamente , Enfermedades del Sistema Nervioso Periférico/patología , Animales , Humanos , Neuronas/efectos de los fármacos , Neuronas/patologíaRESUMEN
The serine/cysteine protease inhibitor phenylmethylsulfonyl fluoride (PMSF) has been used both to promote and to protect against neuropathic events of organophosphorus-induced delayed neuropathy (OPIDN) in hens (Veronesi and Padilla, 1985; Pope and Padilla, 1990; Lotti et al., 1991; Pope et al., 1993; Randall et al., 1997). This study is the first to expand upon this work by using high resolution microscopy provided by epoxy resin embedding and thin sectioning to evaluate neuropathological manifestations of promotion and protection, and to correlate them with associated clinical modifications. To evaluate dose-related effects of OPIDN, single phenyl saligenin phosphate (PSP) dosages of 0.5, 1.0, or 2.5 mg/kg were administered to adult hens. PMSF (90 mg/kg) was given either 4 hours after (for promotion) or 12 hours prior to (for protection) PSP administration. Clinical signs and pathologic changes in the biventer cervicis nerve, which is uniquely sensitive to OPIDN (El-Fawal et al., 1988), were monitored. PSP alone, 2.5 mg/kg, caused severe OPIDN (terminal clinical score 7.5 +/- 1.0 [0-8 scale]; neuropathology score 2.7 +/- 0.3 [0-4 scale, based on myelinated fiber degeneration]). PMSF given 12 hours prior to PSP gave complete protection (clinical and neuropathology scores of 0; p<0.0001 compared to PSP alone). Signs and lesions of OPIDN were absent following 0.5 mg/kg PSP alone, but PMSF given 4 hours after PSP potentiated its neurotoxic effects (all hens had clinical scores of 4.0 and the average neuropathology score was 3.5 +/- 0.3; p<0.0001 compared to PSP alone). Although quantitative differences were noted, qualitative differences among nerves from hens with OPIDN were not evident, either with light or electron microscopy. At the time of sacrifice, there was a statistically linear relationship (r2 = 0.76) between the clinical scores on the last day of observation and the neuropathology scores (p<0.0001). This study demonstrates that the degree of peripheral nerve myelinated fiber degeneration correlates with clinical deficits in PMSF-induced potentiation of and protection against OPIDN.
Asunto(s)
Pollos/fisiología , Enfermedades del Sistema Nervioso/inducido químicamente , Compuestos Organofosforados/toxicidad , Fluoruro de Fenilmetilsulfonilo/toxicidad , Inhibidores de Proteasas/toxicidad , Animales , Encéfalo/patología , Encéfalo/ultraestructura , Femenino , Degeneración Nerviosa/inducido químicamente , Degeneración Nerviosa/patología , Fibras Nerviosas/patología , Fibras Nerviosas/ultraestructura , Enfermedades del Sistema Nervioso/patología , Adhesión en Plástico , Médula Espinal/patología , Médula Espinal/ultraestructura , Fijación del TejidoRESUMEN
This paper reviews the characteristics of organophosphate-induced delayed neurotoxicity, its mechanism, lesions, species sensitivities and structure activity-relationships as they relate to the class of compounds known as triaryl phosphates. The triaryl phosphates have been widely used in commerce for over thirty years as flame retardants in fluids and plastics. Concern has been raised regarding their potential to cause organophosphate-induced delayed neurotoxicity (OPIDN), due to structural similarities to the potent neurotoxicant, tri-ortho cresyl phosphate (TOCP). Based on research on many pure isomers, Johnson (1975a, 1975b) found that certain structural features are required for a triaryl phosphate to react with the enzyme, neuropathy target enzyme (NTE), in a manner which induces OPIDN. Results of acute hen OPIDN studies, the experimental model of choice, support his findings as regards the structure-activity relationships for commercial triaryl phosphates. Thus, standard acute hen OPIDN studies on triphenyl phosphate and butylated triaryl phosphates fail to demonstrate a potential to elicit OPIDN by these products after a single dose. Studies on the mixed isopropyl phenyl phosphates indicate that, while some are neurotoxic, they are much less potent than tricresyl phosphate (TCP) and TOCP in the induction of OPIDN. Most commercial isopropylated triaryl phosphates lacked the potential to induce acute OPIDN using a limit dose of 2000 mg/kg. Although in early studies these compounds appeared to be neurotoxic, they were generally tested at excessively high doses, often exceeding 10,000 mg/kg in acute hen OPIDN studies. In contrast to the isopropylated and butylated triaryl phosphate products, TCP, and especially its ortho substituted isomer, TOCP, were found to be neurotoxic in both acute and subchronic hen OPIDN studies. Recent advances in the synthesis of commercial TCP products have resulted in products with reduced neurotoxic potential (McCormick et al, 1993). As an example, when 3% TCP in aviation oil was dosed acutely at 5000 mg/kg, or for 90 days at 1000 mg/kg/day, no delayed neurotoxicity was noted (Daughtrey et al., 1990, 1996). These data are indicative of the safety of these aviation lubricants at use levels currently employed.