RESUMO
Anticorpos monoclonais (AcM) para rotavírus bovino foram caracterizados para sua aplicação como ferramenta de diagnóstico, utilizando-se as técnicas de isotipificação, dot-blot, western-blot, imunofluorescência indireta (IFI) e ELISA de captura. A caracterização imunoquímica demonstrou que os cinco AcM 1G5, 4F7, 1E12, 4F3 e 3C12 foram do isótipo IgG2a. Pela técnica de dot-blot, os AcM 1G5, 4F7, 1E12, 4F3 detectaram antígenos do rotavírus, em diferentes concentrações, e dois AcM (1E12 e 4F3) reconheceram proteínas virais pela técnica de western-blot. Todos os AcM reagiram positivamente na técnica de IFI em cultivo celular e foram capazes de detectar antígeno viral em amostras fecais bovinas e humanas, pela técnica de ELISA de captura. Identificaram-se dois grupos de AcM, um deles formado pelos AcM 4F7, 1E12 e 1G5, para seu possível uso na detecção de antígeno viral em fezes por meio do ELISA de captura ou dot-blot e outro pelos 4F3 e 3C12, que podem ser usados para detectar antígeno viral em culturas de células por meio de IFI.(AU)
This work was carried out to characterize and evaluate five bovine rotavirus, monoclonal antibodies (MAbs), as a diagnosis tool, by isotyping, dot-blot, western-blot, indirect immunofluorescence (IFI) and ELISA techniques. The immunochemistry characterization showed that all five MAbs (4F7, 4F3, 1G5, 1E12 and 3C12) were IgG2a isotype. The dot-blot immunoassay showed that 1G5, 4F7, 1E12 and 4F3 detected viral antigen in different concentrations and two MAbs (1E12 and 4F3) recognized viral proteins by western-blot. All MAbs detected viral antigen in bovine and human fecal samples by capture ELISA technique and viral antigen in infected MA-104 cell culture by IFI. In conclusion, two groups of Mabs were indetified: one with Mabs 4F7, 1E12 and 1G5 showed the best results to detect rotavirus antigen in fecal samples by capture ELISA or dot-blot techniques assay and other with 4F3 and 3C12 which may be used to detect rotavirus antigens in cell culture by IFI. The results showed the potential use of these MAbs as diagnosis tools in diarrheas by rotavirus in bovines.(AU)
Assuntos
Animais , Bovinos , Anticorpos Monoclonais , Rotavirus/imunologia , Rotavirus/isolamento & purificação , Antígenos Virais/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/métodos , Immunoblotting/veterinária , Western Blotting/métodos , Bovinos/imunologiaRESUMO
Anticorpos monoclonais (AcM) para rotavírus bovino foram caracterizados para sua aplicação como ferramenta de diagnóstico, utilizando-se as técnicas de isotipificação, dot-blot, western-blot, imunofluorescência indireta (IFI) e ELISA de captura. A caracterização imunoquímica demonstrou que os cinco AcM 1G5, 4F7, 1E12, 4F3 e 3C12 foram do isótipo IgG2a. Pela técnica de dot-blot, os AcM 1G5, 4F7, 1E12, 4F3 detectaram antígenos do rotavírus, em diferentes concentrações, e dois AcM (1E12 e 4F3) reconheceram proteínas virais pela técnica de western-blot. Todos os AcM reagiram positivamente na técnica de IFI em cultivo celular e foram capazes de detectar antígeno viral em amostras fecais bovinas e humanas, pela técnica de ELISA de captura. Identificaram-se dois grupos de AcM, um deles formado pelos AcM 4F7, 1E12 e 1G5, para seu possível uso na detecção de antígeno viral em fezes por meio do ELISA de captura ou dot-blot e outro pelos 4F3 e 3C12, que podem ser usados para detectar antígeno viral em culturas de células por meio de IFI.
This work was carried out to characterize and evaluate five bovine rotavirus, monoclonal antibodies (MAbs), as a diagnosis tool, by isotyping, dot-blot, western-blot, indirect immunofluorescence (IFI) and ELISA techniques. The immunochemistry characterization showed that all five MAbs (4F7, 4F3, 1G5, 1E12 and 3C12) were IgG2a isotype. The dot-blot immunoassay showed that 1G5, 4F7, 1E12 and 4F3 detected viral antigen in different concentrations and two MAbs (1E12 and 4F3) recognized viral proteins by western-blot. All MAbs detected viral antigen in bovine and human fecal samples by capture ELISA technique and viral antigen in infected MA-104 cell culture by IFI. In conclusion, two groups of Mabs were indetified: one with Mabs 4F7, 1E12 and 1G5 showed the best results to detect rotavirus antigen in fecal samples by capture ELISA or dot-blot techniques assay and other with 4F3 and 3C12 which may be used to detect rotavirus antigens in cell culture by IFI. The results showed the potential use of these MAbs as diagnosis tools in diarrheas by rotavirus in bovines.
Assuntos
Animais , Bovinos , Anticorpos Monoclonais , Antígenos Virais/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Immunoblotting/veterinária , Rotavirus/imunologia , Rotavirus/isolamento & purificação , Western Blotting/métodos , Bovinos/imunologiaRESUMO
BACKGROUND: The risk of skin cancer is correlated with skin colour. Pigmentation protects against the effects of UV radiation. AIM: To study skin, eyes and hair colour in Chilean teenagers. MATERIAL AND METHODS: The constitutive skin, hair and eyes colour and the presence of freckles was studied in 716 teenagers (416 females) of a low socio economical level and in 307 teenagers (155 females) of a high socio economical level. RESULTS: The proportion of foreign surnames was higher in the high stratum and we only found aboriginal surnames in the low stratum. The females of the lower stratum presented lighter skin than males. This difference was not observed in the higher stratum. We did not find significant differences in the eye colour between sexes, however, adolescents from the high stratum presented lighter eye colour. Females had lighter hair colour than males in both strata, also, we found lighter hair colour in the high stratum. We did not find significant differences in the presence of freckles between strata, but, the proportion of females with freckles was higher than that of males in both strata. CONCLUSIONS: Our results may identify different groups of people, within the Chilean population, with different susceptibility to the effects of ultraviolet radiation.
Assuntos
Humanos , Masculino , Feminino , Adolescente , Caracteres Sexuais , Cor de Cabelo , Melanose/epidemiologia , Pigmentação da Pele , Chile/epidemiologia , Classe Social , Fatores de Risco , Neoplasias Cutâneas/etiologia , Raios Ultravioleta/efeitos adversosRESUMO
The correction of groin hernias has increasingly been performed by transperitoneal videolaparoscopic methods, using a polypropylene mesh. This could lead to an increased incidence of adhesion formation. The incidence of adhesions induced by mesh placement and the influence of reperitonization was observed on 40 male adult Wistar rats. Adhesions were significantly more common on the groups in which the prosthesis was placed (59% vs 95%; p = 0.01), as well as on the groups in which reperitonization was performed (58% vs 100%; p = 0.03). The results suggest that polypropylene mesh placement and reperitonization are each independent factors in the induction of adhesions formation.