Resultado da pesquisa (5)
Termo utilizado na pesquisa Rosa J.C.
#1 - Isolation and culture of neurons and neurospheres from chicken brain cortex, 33(Supl.1):45-50
Abstract in English:
ABSTRACT.- Crepaldi C.R., Merighe G.K.F., Laure H.J., Rosa J.C., Meirelles F.V. & Cerqueira César M. 2013. [Isolation and culture of neurons and neurospheres from chicken brain cortex.] Isolamento e cultivo de neurônios e neuroesferas de córtex cerebral aviar. Pesquisa Veterinária Brasileira 33(Supl.1):45-50. Laboratório de Neurociência e Proteômica, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Av. Duque de Caxias Norte 225, Pirassununga, SP 13635-900, Brazil. E-mail: mccesar@usp.br
Cell culture methods are used for studies of protein interactions in neural cells, helping to detect the interactome of proteins linked to generation of central nervous system diseases. For this reason, neural cells and neurospheres isolated from cortical chicken brain are a current model for studies of neurological diseases, such as epilepsy. Chicken brain has key characteristics on its proteome, with a differential expression of proteins linked to energy metabolism, some of them (VDAC 1 and VDAC 2) play an important role in understanding mechanism of refractory epilepsy. Using the methods described, we found neurospheres, in which cells grow in structures with the ideal diameter of 100-200µm within seven days after isolation. Neurospheres differentiation was obtained after adhesion of these cells to surfaces coated with poly-D-Lysine, detected by migration of fibers inside them. Unlike neurospheres, neurons extended neurites after 11 days of isolation. Here we describe a method to isolate and culture neurons and neurospheres from chicken cerebral cortex. Such “in vitro” model can be utilized on studies of neuronal protein differential expression and interaction. Cultures of isolated neurons represent an accessible model on studies of apoptosis and channel blockers of key proteins linked to brain metabolism.
Abstract in Portuguese:
RESUMO.- Crepaldi C.R., Merighe G.K.F., Laure H.J., Rosa J.C., Meirelles F.V. & Cerqueira César M. 2013. [Isolation and culture of neurons and neurospheres from chicken brain cortex.] Isolamento e cultivo de neurônios e neuroesferas de córtex cerebral aviar. Pesquisa Veterinária Brasileira 33(Supl.1):45-50. Laboratório de Neurociência e Proteômica, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Av. Duque de Caxias Norte 225, Pirassununga, SP 13635-900, Brazil. E-mail: mccesar@usp.br
Métodos de cultivo celular são convenientes na realização de análises funcionais de alterações/interações protéicas das células neuronais, auxiliando a decifrar o interactoma de proteínas chaves na neurogênese de doenças do Sistema Nervoso Central. Por esse motivo, culturas de neurônios e neuroesferas isolados do córtex cerebral aviar representam um modelo acessível para o estudo de diversas doenças neurológicas, tal como a epilepsia. A espécie aviar apresenta peculiaridades em seu proteoma neuronal, visto a presença de uma expressão diferenciada de proteínas chaves no metabolismo energético cerebral, algumas destas (VDAC1 e VDAC2) desempenham papel importante na compreensão do mecanismo da epilepsia refratária. A metodologia estabelecida no presente estudo obteve cultivo de neuroeferas, onde as células cresceram tipicamente em aglomerados atingindo, dentro de 7 dias, o diâmetro ideal de 100-200 µm. A diferenciação celular das neuroesferas foi obtida após a aderência destas às placas tratadas com poli-D-lisina, evidenciada pela migração de fibras do interior da neuroesfera. Ao contrário das neuroesferas, os neurônios em cultivo extenderam seus neuritos após 11 dias de isolamento. Tal modelo in vitro pode ser utilizado com sucesso na identificação das variáveis neuroproteômicas, propiciando uma avaliação global das alterações dinâmicas e suas interações protéicas. Tal modelo pode ter aplicações em estudos dos efeitos de indutores da morte celular e bloqueadores de canais de membrana mitocondriais em proteínas chaves do metabolismo energético cerebral.
#2 - Diagnóstico de raiva no Rio Grande do Sul, Brasil, de 1985 a 2007, p.515-520
Abstract in English:
ABSTRACT.- Teixeira T.F., Holz C.L., Caixeta S.P.M.B., Dezen D., Cibulski S.P., Silva J.R., Rosa J.C.A., Schmidt E., Ferreira J.C., Batista H.B.C.R., Caldas E., Franco A.C. & Roehe P.M. 2008. [Rabies diagnosis in the state of Rio Grande do Sul, Brazil, from 1985 to 2007.] Diagnóstico de raiva no Rio Grande do Sul, Brasil, de 1985 a 2007. Pesquisa Veterinária Brasileira 28(10):515-520. Instituto de Pesquisas Veterinárias Desidério Finamor, Fepagro-Saúde Animal, Cx. Postal 2076, Porto Alegre, RS 90001-970, Brazil. E-mail: proehe@ufrgs.br
The results of 23 years of rabies diagnosis carried out at the Veterinary Research Institute Desidério Finamor, in the state of Rio Grande do Sul, RS, Brazil, are reported. From 1985 to 2007, a total of 23.460 specimens were examined, corresponding to 95% of the total number of samples submitted to rabies laboratory diagnosis notified within the state. Diagnostic methods included standard techniques such as the fluorescent antibody test (FAT) and mouse inoculation test (MIT). No cases of human rabies occurred in the period. Rabies virus (RV) was detected in 739 specimens (3.1%), from which 656 (88.7%) were from cattle. The virus was also identified in specimens from 23 dogs (3.1%), 21 horses (2.9%), 29 bats (4.0%), 4 cats (0.5%), 3 sheep (0.4%), 2 pigs (0.27%) and a wild animal of undetermined species (0.13%). The last case of rabies associated with a canine variant was diagnosed in 1988. Two cases of rabies associated with bat variant viruses were reported, in a domestic cat (2001) and in a dog (2007). In cattle, a marked tendency to a decrease in the number of cases was detected in the examined period. In contrast, an increase in the number of cases in haematophagous as well as in non haematophagous bats is noticed. However, as the number of bat specimens submitted for diagnosis has increased, this finding most likely reflects a higher degree of awareness on the possible role for bats in the rabies transmission cycle, rather than any particular changes on the virus or its hosts.
Abstract in Portuguese:
ABSTRACT.- Teixeira T.F., Holz C.L., Caixeta S.P.M.B., Dezen D., Cibulski S.P., Silva J.R., Rosa J.C.A., Schmidt E., Ferreira J.C., Batista H.B.C.R., Caldas E., Franco A.C. & Roehe P.M. 2008. [Rabies diagnosis in the state of Rio Grande do Sul, Brazil, from 1985 to 2007.] Diagnóstico de raiva no Rio Grande do Sul, Brasil, de 1985 a 2007. Pesquisa Veterinária Brasileira 28(10):515-520. Instituto de Pesquisas Veterinárias Desidério Finamor, Fepagro-Saúde Animal, Cx. Postal 2076, Porto Alegre, RS 90001-970, Brazil. E-mail: proehe@ufrgs.br
The results of 23 years of rabies diagnosis carried out at the Veterinary Research Institute Desidério Finamor, in the state of Rio Grande do Sul, RS, Brazil, are reported. From 1985 to 2007, a total of 23.460 specimens were examined, corresponding to 95% of the total number of samples submitted to rabies laboratory diagnosis notified within the state. Diagnostic methods included standard techniques such as the fluorescent antibody test (FAT) and mouse inoculation test (MIT). No cases of human rabies occurred in the period. Rabies virus (RV) was detected in 739 specimens (3.1%), from which 656 (88.7%) were from cattle. The virus was also identified in specimens from 23 dogs (3.1%), 21 horses (2.9%), 29 bats (4.0%), 4 cats (0.5%), 3 sheep (0.4%), 2 pigs (0.27%) and a wild animal of undetermined species (0.13%). The last case of rabies associated with a canine variant was diagnosed in 1988. Two cases of rabies associated with bat variant viruses were reported, in a domestic cat (2001) and in a dog (2007). In cattle, a marked tendency to a decrease in the number of cases was detected in the examined period. In contrast, an increase in the number of cases in haematophagous as well as in non haematophagous bats is noticed. However, as the number of bat specimens submitted for diagnosis has increased, this finding most likely reflects a higher degree of awareness on the possible role for bats in the rabies transmission cycle, rather than any particular changes on the virus or its hosts.
#3 - Caracterização antigênica e molecular de oito amostras do virus da doença de Aujeszky isoladas no estado do Rio Grande do Sul em 2003, p.21-24
Abstract in English:
D'Ávila da Silva A., Sortica V.A., Braga A.C., Spilki F.R., Franco A.C., Esteves P.A., Rijsewijk F., Rosa J.C.A., Batista H.B.C.R., Oliveira A.P. & Roehe P.M. 2005. [Antigenic and molecular characterization of eight samples of Aujeszky’s disease virus isolated in the state of Rio Grande do Sul, Brazil, in 2003.] Caracterização antigênica e molecular de oito amostras do virus da doença de Aujeszky isoladas no estado do Rio Grande do Sul em 2003. Pesquisa Veterinária Brasileira 25(1):21-24. Instituto de Pesquisas Veterinárias Desidério Finamor (IPVDF), Fepagro Saúde Animal, Estrada do Conde 6000, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@ufrgs.br
Pseudorabies or Aujeszky’s disease (AD), caused by pseudorabies virus (PRV) is a major concern in swine production. In the state of Rio Grande do Sul, Brazil, AD was only detected in 1954, in cattle. In 2003 two outbreaks of encephalitis occurred on the northern region of the state, close to the border with the state of Santa Catarina. Pseudorabies virus (PRV) was isolated from distinct farms within the region and subjected to antigenic and genomic analyses. These isolates were compared with prototype strains NIA-3 and NP. Antigenic characterization with a panel of monoclonal antibodies (Mabs) directed to viral glycoproteins (gB, gC, gD and gE,) was performed by an imunoperoxidase monolayer assay (IPMA) on infected cell monolayers. Genomic characterization was carried out by restriction enzyme analysis (REA) of the whole DNA viral genome with Bam HI. The antigenic profile of the eight isolates from Rio Grande do Sul as well as strains NIA-3 and NP were similar. REA analysis revealed that all isolates from Rio Grande do Sul displayed a genomic type II arrangement, a genotype often found in other outbreaks of AD previously reported in other Brazilian states. The results obtained suggest that the eight isolates examined here were similar.
Abstract in Portuguese:
D'Ávila da Silva A., Sortica V.A., Braga A.C., Spilki F.R., Franco A.C., Esteves P.A., Rijsewijk F., Rosa J.C.A., Batista H.B.C.R., Oliveira A.P. & Roehe P.M. 2005. [Antigenic and molecular characterization of eight samples of Aujeszky’s disease virus isolated in the state of Rio Grande do Sul, Brazil, in 2003.] Caracterização antigênica e molecular de oito amostras do virus da doença de Aujeszky isoladas no estado do Rio Grande do Sul em 2003. Pesquisa Veterinária Brasileira 25(1):21-24. Instituto de Pesquisas Veterinárias Desidério Finamor (IPVDF), Fepagro Saúde Animal, Estrada do Conde 6000, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@ufrgs.br
Pseudorabies or Aujeszky’s disease (AD), caused by pseudorabies virus (PRV) is a major concern in swine production. In the state of Rio Grande do Sul, Brazil, AD was only detected in 1954, in cattle. In 2003 two outbreaks of encephalitis occurred on the northern region of the state, close to the border with the state of Santa Catarina. Pseudorabies virus (PRV) was isolated from distinct farms within the region and subjected to antigenic and genomic analyses. These isolates were compared with prototype strains NIA-3 and NP. Antigenic characterization with a panel of monoclonal antibodies (Mabs) directed to viral glycoproteins (gB, gC, gD and gE,) was performed by an imunoperoxidase monolayer assay (IPMA) on infected cell monolayers. Genomic characterization was carried out by restriction enzyme analysis (REA) of the whole DNA viral genome with Bam HI. The antigenic profile of the eight isolates from Rio Grande do Sul as well as strains NIA-3 and NP were similar. REA analysis revealed that all isolates from Rio Grande do Sul displayed a genomic type II arrangement, a genotype often found in other outbreaks of AD previously reported in other Brazilian states. The results obtained suggest that the eight isolates examined here were similar.
#4 - Monoclonal antibody differentiation between bovine herpesviruses type 1 and 5, 17(1):41-44
Abstract in English:
ABSTRACT.- Roehe P.M., Silva T.C., Nardi N.B., Oliveira L.G. & Rosa J.C.A. 1997. [Monoclonal antibody differentiation between bovine herpesviruses type 1 and 5.] Diferenciação entre os vírus da rinotraqueíte infecciosa bovina (BHV-1) e herpesvírus da encefalite bovina (BHV-5) com anticorpos monoclonais. Pesquisa Veterinária Brasileira 17(1):41-44. Centro de Pesquisas Veterinárias Desidério Finamor, Caixa Postal 2076, Porto Alegre, RS 90001-970, Brazil.
Bovine Herpesviruses (BHV) type 1 (BHV-1) and type 5 (BHV-5) were analysed by immunoperoxidase staining with a panei of monoclonal antibodies (Mabs) prepared against BHV antigens. One of the Mabs recognized ali BHV isolates tested. The remainder four mabs recognized only BHV-1 samples, including standard laboratory strains. Ali isolates associated with clinical cases of encephalitis (BHV-5) displayed a pattern of reactivity distinct from that of viruses isolated from syndromes associated with BHV-1 infections. The results obtained indicate that such Mabs allowed the differentiation between BHV-1 and BHV-5, with a perfect correlation between the clinical pictures and the patterns of reactivity in vitro.
Abstract in Portuguese:
SINOPSE.- Roehe P.M., Silva T.C., Nardi N.B., Oliveira L.G. & Rosa J.C.A. 1997. [Monoclonal antibody differentiation between bovine herpesviruses type 1 and 5.] Diferenciação entre os vírus da rinotraqueíte infecciosa bovina (BHV-1) e herpesvírus da encefalite bovina (BHV-5) com anticorpos monoclonais. Pesquisa Veterinária Brasileira 17(1):41-44. Centro de Pesquisas Veterinárias Desidério Finamor, Caixa Postal 2076, Porto Alegre, RS 90001-970, Brazil.
Amostras de herpesvírus bovinos (BHV) tipo 1 (Virus da Rinotraqueíte Infecciosa BovinaNulvovaginite Pustular Infecciosa; BHV-1) e tipo 5 (Herpesvírus da Encefalite Bovina; BHV-5) tiveram seu perfil de reatividade analisado em testes de imunoperoxidase frente a um painel composto por cinco anticorpos monoclonais (AcM) produzidos contra antígenos de BHV-1. Um dos AcM reconheceu todas as amostras de BHV examinadas. Os quatro AcM restantes reconheceram somente amostras de BHV-1. Todas as amostras isoladas de casos de encefalites (BHV-5) apresentaram um padrão de reação distinto daquelas isoladas de outros síndromes associados à infecção pelo BHV-1. Os resultados obtidos indicam que os AcM avaliados permitem a diferenciação entre amostras de BHV-1 e BHV-5, havendo perfeita correlação entre os quadros clínicos observados com os perfis de reatividade obtidos in vitro.
#5 - Rotavirus in swine in the southem region of Brazil
Abstract in English:
The occurrence of rotavirus infection was studied in faeces of diarrhoeic swine in 32 farms of 22 municipalities from the states of Rio Grande do Sul and Santa Catarina. From a total number of 231 faecal samples collected from piglets aging 1 to 60 days, 43 (18.6%) were positive by polyacrylamide gel electrophoresis (PAGE), and 42 (17.7%) were positive by an enzyme immunoassay (ELISA). With the exception of 1 sample, the electropherotypes observed were consistent with the description of classical group A rotavirus. in 5 samples rotavirus was detected by ELISA and not by PAGE, whereas the inverse occurred in 7 samples. Studies with faecal dilutions of positive faeces demonstrated that the discrepancies probably were not due to differences in virus concentration. The presence of IgG was confirmed in one sample PAGE positive - ELISA negative, suggesting the presence of immune complexes that may have interfered in the results obtained. Bacteriological and parasitological examinations in 75 samples showed the presence of enteropathogenic Escherichia coli in 8 (10.7%) samples and Treponema hyodysenteriae in 6 (8%). in no sample were parasites detected. In only one sample E. coli and rotavirus were detected together, suggesting low incidence of the association bacteria-viruses. However, the severity of the outbreak where association was detected was greater than in the others. The age distribution of rotavirus infection in the piglets showed that 90% occurred in animals between 15 and 30 days old.
Abstract in Portuguese:
A ocorrência de infecção por rotavirus foi estudada em granjas de suínos acometidos de diarréia em 32 propriedades de 22 municípios do Rio Grande do Sul e Santa Catarina. De um total de 231 amostras fecais coletadas de leitões na faixa etária de 1 a 60 dias, 43 (18,6%) foram positivas pela técnica de eletroforese em gel de poliacrilamida (PAGE) e 42 (17,7%) pelo ensaio imúnoenzimático (ELISA). Com a exceção de um caso, os eletroferotipos observados foram consistentes com a descrição dos rotavirus "clássicos" do grupo A. Em 5 amostras o rotavirus foi detectado por ELISA e não por PAGE, enquanto que a situação inversa ocorreu em 7 casos. Estudos com diluições de fezes positivas demonstraram que as discrepâncias observadas provavelmente não foram devidas a diferenças nas concentrações de vírus. A presença de IgG foi verificada em uma amostra PAGE positiva ELISA negativa, sugerindo a presença de complexos imunes que poderiam ser a causa de discrepâncias entre os métodos de diagnóstico. Exames bacteriológicos e parasitológicos em 75 amostras revelaram a presença de Escherichia coli enteropatogênica em 8 (10, 7%) e Treponema hyodysenteriae em 6 (8%). Em nenhum caso foram detectados parasitos. Em somente um caso E. coli foi encontrada em associação com rotavirus, que clinicamente mostrou maior gravidade do que nos demais. A distribuição da infecção por faixas etárias revelou que 79% dos casos foram verificados em animais de 15 a 30 dias.
