Made available in DSpace on 2014-06-11T19:31:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-05-16Bitstream added on 2014-06-13T20:22:04Z : No. of bitstreams: 1 zuanaze_rcc_dr_botfmvz.pdf: 2365698 bytes, checksum: 5ed70ccdd25955ad8f5b2e015a21dec7 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação para o Desenvolvimento da UNESP (FUNDUNESP) / O monofluoroacetato de sodio (MFAS) ou composto 1080 e um rodenticida que foi amplamente utilizado para o controle de roedores e predadores domesticos, apos sua descoberta em 1945. Este potente rodenticida age bloqueando o ciclo de Krebs por acao do seu metabolito toxico, o fluorocitrato, e como consequencia, inibe a resposta celular e a producao de ATP. O MFAS foi proibido por lei no Brasil e em diversos paises, mas seu uso indiscriminado continua causando diversos casos de intoxicacoes potencialmente fatais no homem e animais domesticos, principalmente em caes e gatos, representadas por alteracoes neurologicas e cardiacas. Pretendeu-se com este estudo caracterizar o perfil hematologico e bioquimico, as alteracoes histopatologicas e toxicologicas de gatos intoxicados experimentalmente com MFAS, com o objetivo de determinar metodos diagnosticos eficazes. Para tanto, foram utilizadas amostras de sangue de 16 gatos intoxicados experimentalmente com 0,45mg/kg de MFAS, por via oral. Estas amostras foram colhidas por puncao jugular e analisadas quanto ao perfil hematologico, bioquimica serica e analises toxicologicas em cromatografia liquida de alta eficiencia (CLAE), para deteccao e quantificacao do MFAS no soro dos animais. Avaliaram-se tambem as lesoes macro e microscopicas dos animais intoxicados que vieram a obito. Observaram-se leucopenia e trombocitopenia transitorias; hiperglicemia, aumento das enzimas musculares creatinoquinase (CK) e creatinoquinase fracao cardiaca (CK-MB); observaram-se tambem hipocalemia, hipofosfatemia e hipomagnesemia. Os achados macroscopicos e histopatologicos demonstraram lesoes caracteristicas de processos isquemicos e as analises toxicologicas demonstraram um metodo diagnostico simples e eficiente... / Sodium monofluoroacetate (SMFAC) or 1080 compound is a potent rodenticide, largely used since 1945 for rodent and domestic pest control. The toxic effects of SMFAC are caused by fluorocitrato, a toxic metabolite, which has a competitive action with aconitase enzyme, leading to citrate accumulation and interferes in energy production by Krebs cycle blockade. In Brazil, although prohibited by law, there is illegal use, keep causing intoxication in children and domestic animals, specially dogs and cats. The most common intoxication clinical signs are from the cardiac and neurological alterations. In the present study, 16 domestic cats were intoxicated with oral doses of monofluoroacetate (0.45mg/kg). The hematologic and biochemical profiles, and histophatological and blood serum toxicological analysis by high performance liquid chromatography (HPLC) were made to looking for a efficient diagnosis methods. The hematologic profile showed transitory leucopenia and trombocitopenia; the biochemical profile presented hiperglycemia, increase of creatinoquinase enzyme (CK) and creatinoquinase cardiac fraction (CK-MB), hypokalemia and hypophosfatemia were observed. The macrocospic and histopathological findings showed lesions characteristic of degenerative and ischemic processes. The toxicological analysis was shown to be a simple and efficient diagnostic method. SMFAC was detected in 75% of the serum samples analysed, and it was verified an average concentration of 0.32 ìg/mL in them; 4.81% of the serum samples did not show metabolized SMFAC six hours after the induced intoxication of the animals used in this study.

Orientador: Michiko Sakate / Banca: Noeme Sousa Rocha / Banca: Manoel Lima de Menezes / Banca: Mara Regina Stipp Balarin / Banca: Silvana Lima Gorniak / Resumo: O monofluoroacetato de sodio (MFAS) ou composto 1080 e um rodenticida que foi amplamente utilizado para o controle de roedores e predadores domesticos, apos sua descoberta em 1945. Este potente rodenticida age bloqueando o ciclo de Krebs por acao do seu metabolito toxico, o fluorocitrato, e como consequencia, inibe a resposta celular e a producao de ATP. O MFAS foi proibido por lei no Brasil e em diversos paises, mas seu uso indiscriminado continua causando diversos casos de intoxicacoes potencialmente fatais no homem e animais domesticos, principalmente em caes e gatos, representadas por alteracoes neurologicas e cardiacas. Pretendeu-se com este estudo caracterizar o perfil hematologico e bioquimico, as alteracoes histopatologicas e toxicologicas de gatos intoxicados experimentalmente com MFAS, com o objetivo de determinar metodos diagnosticos eficazes. Para tanto, foram utilizadas amostras de sangue de 16 gatos intoxicados experimentalmente com 0,45mg/kg de MFAS, por via oral. Estas amostras foram colhidas por puncao jugular e analisadas quanto ao perfil hematologico, bioquimica serica e analises toxicologicas em cromatografia liquida de alta eficiencia (CLAE), para deteccao e quantificacao do MFAS no soro dos animais. Avaliaram-se tambem as lesoes macro e microscopicas dos animais intoxicados que vieram a obito. Observaram-se leucopenia e trombocitopenia transitorias; hiperglicemia, aumento das enzimas musculares creatinoquinase (CK) e creatinoquinase fracao cardiaca (CK-MB); observaram-se tambem hipocalemia, hipofosfatemia e hipomagnesemia. Os achados macroscopicos e histopatologicos demonstraram lesoes caracteristicas de processos isquemicos e as analises toxicologicas demonstraram um metodo diagnostico simples e eficiente...(Resumo completo, clicar acesso eletrõnico abaixo) / Abstract: Sodium monofluoroacetate (SMFAC) or 1080 compound is a potent rodenticide, largely used since 1945 for rodent and domestic pest control. The toxic effects of SMFAC are caused by fluorocitrato, a toxic metabolite, which has a competitive action with aconitase enzyme, leading to citrate accumulation and interferes in energy production by Krebs cycle blockade. In Brazil, although prohibited by law, there is illegal use, keep causing intoxication in children and domestic animals, specially dogs and cats. The most common intoxication clinical signs are from the cardiac and neurological alterations. In the present study, 16 domestic cats were intoxicated with oral doses of monofluoroacetate (0.45mg/kg). The hematologic and biochemical profiles, and histophatological and blood serum toxicological analysis by high performance liquid chromatography (HPLC) were made to looking for a efficient diagnosis methods. The hematologic profile showed transitory leucopenia and trombocitopenia; the biochemical profile presented hiperglycemia, increase of creatinoquinase enzyme (CK) and creatinoquinase cardiac fraction (CK-MB), hypokalemia and hypophosfatemia were observed. The macrocospic and histopathological findings showed lesions characteristic of degenerative and ischemic processes. The toxicological analysis was shown to be a simple and efficient diagnostic method. SMFAC was detected in 75% of the serum samples analysed, and it was verified an average concentration of 0.32 ìg/mL in them; 4.81% of the serum samples did not show metabolized SMFAC six hours after the induced intoxication of the animals used in this study. / Doutor

Microtransplantation of mammalian neurolemma into Xenopus laevis oocytes has been used to study ion channels in terms of their structure and function in the central nervous system. Use of microtransplanted neurolemma is advantageous in that tissue can be obtained from various sources, ion channels and receptors are present in their native configuration and they can be used to evaluate numerous channelpathies caused by environmental toxicants. Here we show that Xenopus oocytes injected with fragments of rat brain neurolemma successfully express functional native ion channels that are assembled in their own plasma membrane. Using a high throughput two electrode voltage clamp (TEVC) electrophysiological system, currents that were sensitive to tetrodotoxin (TTX), omega-conotoxin MVIIC, and tetraethylammonium (TEA) were detected, indicating the presence of multiple voltage-sensitive ion channels (voltage-sensitive sodium, calcium and potassium channels, respectively). In this current research, a “proof-of-principle” experiment was conducted where TTX-sensitive voltage-sensitive sodium channel (VSSC) currents were measured. VSSCs are a well-established site of action for 1,1,1-trichloro-2,2-di(4-chlorophenyl)ethane (DDT) but not for its non-toxic metabolite 1,1-bis-(4-chlorophenyl)-2,2-dichloroethene (DDE). A differential sensitivity of DDT versus DDE on TTX-sensitive sodium current in neurolemma-injected oocytes was determined. DDT elicited an increase in depolarization-dependent, TTX-sensitive sodium current while DDE had no significant effect. Additionally, DDT resulted in a slowing of sodium channel inactivation kinetics whereas DDE has no similar effect. These results are consistent with the findings obtained using heterologous expression of single isoforms of rat brain VSSCs by injecting cRNA into Xenopus oocytes. By demonstrating the classic structural activity relationship of DDT and DDE on mammalian voltage-gated sodium channels isolated in rat brain neurolemma, this study supports the use of automated high-throughput electrophysiology to study the effects of various environmental toxicants on multiple mammalian cellular targets. More importantly, using rat brain neurolemma ensures that the proteins of interest have been transcribed and have undergone all the necessary post-translational modifications before they were injected and expressed in the Xenopus oocytes which is not the case for traditional heterologous expression.