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1.
Ci. Rural ; 46(10): 1830-1837, Oct. 2016. ilus, graf, tab
Artigo em Inglês | VETINDEX | ID: vti-29794

Resumo

Wharton's jelly is a source of mesenchymal stem cells (MSCs) that had not yet been tested for bovine embryo production by nuclear transfer (NT). Thus, the objective of this study was to isolate, characterize and test MSCs derived from Wharton's jelly for embryo and pregnancy production by NT in cattle. The umbilical cord was collected during calving and cells derived from Wharton's jelly (WJCs) were isolated by explant and cultured in Dulbecco's Modified Eagle Medium. Skin Fibroblasts (FB) were isolated after 6 months of life. Morphological analysis was performed by bright field and scanning electron microscopy (SEM) during cell culture. Phenotypic and genotypic characterization by flow cytometry, immunocytochemistry, RT-PCR and differentiation induction in cell lineages were performed for WJC. In the NT procedure, oocytes at the arrested metaphase II stage were enucleated using micromanipulators, fused with WJCs or FB and later activated artificially. SEM micrographs revealed that WJCs have variable shape under culture. Mesenchymal markers of MSCs (CD29+, CD73+, CD90+ and CD105+) were expressed in bovine-derived WJC cultures, as evidenced by flow cytometry, immunocytochemistry and RT-PCR. When induced, these cells differentiated into osteocytes, chondrocytes and adipocytes. After classification, the WJCs were used in NT. Blastocyst formation rate by NT with WJCs at day 7 was 25.80±0.03%, similar to blatocyst rate with NT using skin fibroblasts (19.00±0.07%). Pregnancies were obtained and showed that WJCs constitute a new cell type for use in animal cloning.(AU)


A geleia de Wharton é uma fonte de células tronco mesenquimais (CTMs) que ainda não havia sido testada para a produção de embriões bovinos por transferência nuclear (TN). O objetivo deste estudo foi isolar, caracterizar e testar as CTMs derivadas da geleia de Wharton para produção de embriões e gestações por transferência nuclear em bovinos. O cordão umbilical foi coletado durante o nascimento e as células derivadas da geleia de Wharton (CGWs) foram isoladas por explante e cultivadas em Dulbecco's Modified Eagle Medium. Fibroblastos (FB) da pele foram isolados após 6 meses de vida. As análises morfológicas foram realizadas pelas microscopias de campo claro e eletrônica de varredura durante o cultivo celular. Caracterização fenotípica e genotípica por citometria de fluxo, imunocitoquímica, RT-PCR e indução da diferenciação em linhagens celulares foi realizada com as CGWs. No procedimento de TN, ovócitos no estágio de metáfase II foram enucleados usando micromanipuladores, fusionados com CGWs ou FB e então ativados artificialmente. Micrografias de microscopia de varredura revelaram que CGWs tiveram forma variada sob cultivo. Os marcadores mesenquimais de CTMs (CD29+, CD73+, CD90+ and CD105+) foram expressos em cultura de CGWs bovina, como evidenciado por citometria de fluxo, imunocitoquímica e RT-PCR. Quando induzidas, estas células diferenciaram-se em osteócitos, condrócitos e adipócitos. Após classificação, as CGWs foram utilizadas na TN. A taxa de formação de blastocistos por TN com CGWs no sétimo dia de cultivo foi de 25,80±0,03%, similar a produção de blastócitos por TN com fibroblastos de pele (19,00±0,07). Gestações foram obtidas e mostraram que CGWs constituem um novo tipo celular para ser usado na clonagem animal.(AU)


Assuntos
Animais , Bovinos , Células-Tronco Mesenquimais , Geleia de Wharton , Clonagem de Organismos/veterinária , Cordão Umbilical , Embrião de Mamíferos
2.
J. venom. anim. toxins incl. trop. dis ; 21: 22, 31/03/2015. ilus, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-954738

Resumo

Background:This investigation aimed to evaluate the occurrence of some apoptotic features induced by Leptospira interrogans serovar Icterohaemorrhagiae infection in young BALB/c mice during 2, 4, 7, 10, 14 and 21 days post-infection (dpi).Methods:The animals were euthanized and lung, liver and kidneys were harvested to histopathology analysis and immunohistochemistry to caspase-3 antigen detection was performed.Results:Chromatin condensation in kidney and liver tissues, but not in lung tissue, was observed. Caspase-3 reactive cells, mainly characterized as renal epithelial cells, were detected in the days 14 and 21 at high levels when compared to days 2,4 and 7 (p = 0.025; p <0.05). Lung sections revealed caspase-3 labeled alveolar cells in 10 and 14 days post-infection was higher than observed at 7 days (p = 0.0497; p < 0.05). Liver sections demonstrated reactive cells at a highest level at 14 and 21 days post-infection when comparison to 2,4, 7 and 10 days (p = 0.0069; p<0.05).Conclusions:Our results suggest that infection of L interrogans induce in kidney, liver and lung an activation of apoptosis mediated by caspase-3 dependent pathway in later phases of infectious process.(AU)


Assuntos
Animais , Camundongos , Imuno-Histoquímica , Apoptose , Leptospira interrogans serovar icterohaemorrhagiae , Caspase 3
3.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 21: 1-6, Aug. 11, 2015. ilus, graf
Artigo em Inglês | VETINDEX | ID: vti-759988

Resumo

Background: This investigation aimed to evaluate the occurrence of some apoptotic features induced by Leptospira interrogans serovar Icterohaemorrhagiae infection in young BALB/c mice during 2, 4, 7, 10, 14 and 21 days post-infection (dpi). Methods: The animals were euthanized and lung, liver and kidneys were harvested to histopathology analysis and immunohistochemistry to caspase-3 antigen detection was performed. Results: Chromatin condensation in kidney and liver tissues, but not in lung tissue, was observed. Caspase-3 reactive cells, mainly characterized as renal epithelial cells, were detected in the days 14 and 21 at high levels when compared to days 2,4 and 7 (p = 0.025; p <0.05). Lung sections revealed caspase-3 labeled alveolar cells in 10 and 14 days post-infection was higher than observed at 7 days (p = 0.0497; p < 0.05). Liver sections demonstrated reactive cells at a highest level at 14 and 21 days post-infection when comparison to 2,4, 7 and 10 days (p = 0.0069; p<0.05). Conclusions: Our results suggest that infection of L interrogans induce in kidney, liver and lung an activation of apoptosis mediated by caspase-3 dependent pathway in later phases of infectious process.(AU)


Assuntos
Animais , Camundongos , Apoptose , Leptospira interrogans serovar icterohaemorrhagiae , Doença de Weil/veterinária
4.
J. venom. anim. toxins incl. trop. dis ; 21: 1-6, 31/03/2015. ilus, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1484626

Resumo

Background: This investigation aimed to evaluate the occurrence of some apoptotic features induced by Leptospira interrogans serovar Icterohaemorrhagiae infection in young BALB/c mice during 2, 4, 7, 10, 14 and 21 days post-infection (dpi). Methods: The animals were euthanized and lung, liver and kidneys were harvested to histopathology analysis and immunohistochemistry to caspase-3 antigen detection was performed. Results: Chromatin condensation in kidney and liver tissues, but not in lung tissue, was observed. Caspase-3 reactive cells, mainly characterized as renal epithelial cells, were detected in the days 14 and 21 at high levels when compared to days 2,4 and 7 (p = 0.025; p <0.05). Lung sections revealed caspase-3 labeled alveolar cells in 10 and 14 days post-infection was higher than observed at 7 days (p = 0.0497; p < 0.05). Liver sections demonstrated reactive cells at a highest level at 14 and 21 days post-infection when comparison to 2,4, 7 and 10 days (p = 0.0069; p<0.05). Conclusions: Our results suggest that infection of L interrogans induce in kidney, liver and lung an activation of apoptosis mediated by caspase-3 dependent pathway in later phases of infectious process.


Assuntos
Animais , Camundongos , Apoptose , Doença de Weil/veterinária , Leptospira interrogans serovar icterohaemorrhagiae
5.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 16(1): 178-185, 2010. ilus
Artigo em Inglês | VETINDEX | ID: vti-4225

Resumo

Infection of susceptible ruminants, including domestic cattle (Bos taurus) and American bison (Bison bison), with ovine herpesvirus-2 (OvHV-2) may provoke the fatal vasculitis and lymphoproliferative syndrome, known as malignant catarrhal fever (MCF), reported worldwide. To the best of our knowledge, this is the first report of a clinical case of MCF-like lesions associated with ovine herpesvirus-2 (OvHV-2) infection in young calves (Bos indicus) including central nervous symptoms that occurred in Três Lagoas city, Mato Grosso do Sul state, a border town near São Paulo state, Brazil. The diagnosis was based on typical histological lesions characterized by systemic lymphohistiocytic and fibrinoid vasculitis, confirmed by polymerase chain reaction and subsequent phylogenetic analysis of detected OvHV-2 sequences. This finding indicates that MCF disease is spread among herds concentrated in border areas between Mato Grosso do Sul and São Paulo states.(AU)


Assuntos
Animais , Bovinos , Infecções por Herpesviridae , Febre Catarral Maligna , Ovinos , Bovinos/lesões
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