Assuntos
Colagenases/efeitos adversos , Síndrome de Creutzfeldt-Jakob/etiologia , Diabetes Mellitus Tipo 1/cirurgia , Doença Iatrogênica , Transplante das Ilhotas Pancreáticas/efeitos adversos , Termolisina/efeitos adversos , Coleta de Tecidos e Órgãos/efeitos adversos , Animais , Encéfalo/metabolismo , Síndrome de Creutzfeldt-Jakob/diagnóstico , Síndrome de Creutzfeldt-Jakob/metabolismo , Síndrome de Creutzfeldt-Jakob/transmissão , Humanos , Camundongos , Camundongos Transgênicos , Príons/metabolismo , Medição de Risco , Fatores de RiscoRESUMO
Thermolysin is a zinc metalloprotease that has potential uses in the food industry. The safety of thermolysin has not been demonstrated before, and therefore a series of standard toxicological tests to assess its potential toxicity was undertaken. The thermolysin used in this study was derived from the thermophilic bacterium Geobacillus stearothermophilus, which had undergone chemical mutagenesis to generate strains with increased thermolysin production. Acute toxicity studies in rats and mice showed that thermolysin powder is not acutely toxic with an oral LD50 of more than 18,000 mg/kg (2520 mg/kg thermolysin protein) in rats and more than 24,000 mg/kg (3360 mg/kg protein) in mice. Subchronic feeding studies in rats for 91 days at doses up to 1000 mg/kg (390 mg/kg protein) revealed no significant differences between treated and non-treated groups and a No Observed Effect Level (NOEL) of 1000 mg/kg (390 mg/kg protein) per day was established. Results from genotoxicity tests such as in vitro chromosomal aberration assay and in vivo mouse micronucleus were negative. Allergenicity sequence analysis revealed no evidence suggesting that thermolysin is an allergen. The data presented in this study support the conclusion that thermolysin is safe for use in food production.
Assuntos
Proteínas de Bactérias/efeitos adversos , Aditivos Alimentares/efeitos adversos , Geobacillus stearothermophilus/enzimologia , Termolisina/efeitos adversos , Alérgenos/administração & dosagem , Alérgenos/efeitos adversos , Animais , Antígenos de Bactérias/administração & dosagem , Antígenos de Bactérias/efeitos adversos , Proteínas de Bactérias/administração & dosagem , Linhagem Celular , Cricetinae , Cricetulus , Feminino , Aditivos Alimentares/administração & dosagem , Geobacillus stearothermophilus/imunologia , Masculino , Camundongos , Camundongos Endogâmicos , Testes para Micronúcleos , Testes de Mutagenicidade , Nível de Efeito Adverso não Observado , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Termolisina/administração & dosagem , Testes de Toxicidade Aguda , Testes de Toxicidade SubcrônicaRESUMO
A pharmacokinetic model is described for testing of polymorphonuclear leukocyte (PMN) elastase inhibitors administered by intratracheal or aerosol dosing of hamsters. Acute lung injury, measured as hemorrhage occurring within hours after intratracheal instillation of human PMN elastase, correlated directly with the amount of active enzyme instilled. Hemorrhage began within minutes of elastase instillation, was maximal within 1 h, and remained constant for up to 5 h subsequently. Therefore, inhibition of hemorrhage was used as an assay of the effectiveness of various PMN elastase inhibitors given by the intratracheal route. Lung hemorrhage could also be induced by intratracheal instillation of other elastolytic enzymes, such as thermolysin, and inhibition of hemorrhage was seen only with inhibitors active against the type of elastase used. Methoxysuccinyl-alanyl-alanyl-prolyl-valine-chloromethylketone (MeOSuc-AAPV-CMK), as well as alpha 1-proteinase inhibitor (alpha 1PI) but not tosyl-lysine-chloromethylketone (tosyl-lysine-CMK), inhibited the hemorrhage caused by human PMN elastase, but the specific inhibitors of this enzyme had no effect on thermolysin-induced lung hemorrhage. The duration of activity of these compounds as elastase inhibitors in this model correlated directly with the extent of their persistence in lung lavage fluid as determined by HPLC analysis of compound recovered by bronchoalveolar lavage. (ABSTRACT TRUNCATED AT 250 WORDS)