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Muscle biopsy technique for electrophoresis analysis of fish from the genus Brycon Genet. Mol. Biol.
Hilsdorf,Alexandre; Caneppele,Danilo; Krieger,José Eduardo.
Protein and mitochondrial DNA have been used as molecular markers to assess variability in stock identification studies of fishes. Protein and mtDNA used for electrophoretic analysis are extracted from tissues, which often leads to death of the individuals. In this study, we present a skeletal muscle biopsy procedure to extract mitochondrial DNA that does not require specimen sacrifice. Eighty pirapitinga-do-sul (Brycon opalinus) were biopsied by the present technique, with no mortalities recorded. Total DNA was extracted from muscle and digested by restriction enzymes ApaI and HpaI. The mtDNA fragment patterns were hybridized with 32P-labeled pirapitinga-do-sul mtDNA probes. The described technique is simple and may be useful in protocols requiring tissue...
Tipo: Info:eu-repo/semantics/article
Ano: 1999 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47571999000400014
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Knockdown of E2f1 by RNA interference impairs proliferation of rat cells in vitro Genet. Mol. Biol.
Vasques,Luciana dos Reis; Pujiz,Regiane Simoni; Strauss,Bryan Eric; Krieger,José Eduardo.
E2F1 plays a key role in cell-cycle regulation in mammals, since its transcription factor activity controls genes required for DNA synthesis and apoptosis. E2F1 deregulation is a common feature among different tumor types and can be a major cause of cell proliferation. Thus, blocking E2F1 expression by RNA interference represents a promising therapeutic approach. In this study, the introduction of specific short hairpin RNAs (shRNAs) reduced E2f1 expression by up to 77%, and impaired rat glioma cell proliferation by approximately 70%, as compared to control cells. Furthermore, we investigated the expression of E2f1 target genes, Cyclin A and Cyclin E. Cyclin A was found to be down-regulated, whereas Cyclin E had similar expression to control cells,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: RNAi; ShRNA; E2F1; Proliferation; Cancer.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572010000100005
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Lamellar keratoplasty in rabbits using an allogeneic free omental graft and omentum-derived mesenchymal cells associated with the canine amniotic membrane Ciência Rural
Barros,Séfora Vieira da Silva Gouvêa de; Aldrovani,Marcela; Lacerda,Luciana Cenço Correa de; Horr,Mônica; Marinho,Fábio Andrade; Lima,Tiago Barbalho; Silveira,Camila Pinho Balthazar da; Brito,Fábio Luiz da Cunha; Bassaneze,Vinícius; Flecher,Mayra Cunha; Beltrame,Renato Travassos; Nakamuta,Juliana Sanajotti; Krieger,José Eduardo; Laus,José Luiz; Oliveira,Krishna Duro de.
ABSTRACT: The objective of this research was to evaluate the clinical and microscopic effects in rabbits of lamellar keratoplasty using allogeneic omentum associated with canine amniotic membrane (AM). Rabbits were divided into two groups: one received the allogeneic free omental graft covered with the AM (OM-graft group), while the other received the AM graft containing omental mesenchymal cells (OM-cell group). Clinical signs were evaluated on different postoperative days. After the clinical assessments, the rabbits were euthanized and their corneas were obtained for histopathology and immunohistochemistry (Ki-67, marker for proliferation). Both groups showed chemosis, blepharospasm, eye discharge, hyperemia, and corneal opacity/edema. Neovascularization...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Amniotic membrane; Cornea; Omentum; Progenitor cells; Wound healing..
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782016001001838
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