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Contribution of matrix vesicles and alkaline phosphatase to ectopic bone formation BJMBR
Ciancaglini,P.; Simão,A.M.S.; Camolezi,F.L.; Millán,J.L.; Pizauro,J.M..
Endochondral calcification involves the participation of matrix vesicles (MVs), but it remains unclear whether calcification ectopically induced by implants of demineralized bone matrix also proceeds via MVs. Ectopic bone formation was induced by implanting rat demineralized diaphyseal bone matrix into the dorsal subcutaneous tissue of Wistar rats and was examined histologically and biochemically. Budding of MVs from chondrocytes was observed to serve as nucleation sites for mineralization during induced ectopic osteogenesis, presenting a diameter with Gaussian distribution with a median of 306 ± 103 nm. While the role of tissue-nonspecific alkaline phosphatase (TNAP) during mineralization involves hydrolysis of inorganic pyrophosphate (PPi), it is unclear...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Matrix vesicles; Endochondral ossification; Osseous plate; Alkaline phosphatase; Ectopic mineralization; Calcification.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000500006
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Proteoliposomes as matrix vesicles' biomimetics to study the initiation of skeletal mineralization BJMBR
Simão,A.M.S.; Yadav,M.C.; Ciancaglini,P.; Millán,J.L..
During the process of endochondral bone formation, chondrocytes and osteoblasts mineralize their extracellular matrix by promoting the formation of hydroxyapatite (HA) seed crystals in the sheltered interior of membrane-limited matrix vesicles (MVs). Ion transporters control the availability of phosphate and calcium needed for HA deposition. The lipidic microenvironment in which MV-associated enzymes and transporters function plays a crucial physiological role and must be taken into account when attempting to elucidate their interplay during the initiation of biomineralization. In this short mini-review, we discuss the potential use of proteoliposome systems as chondrocyte- and osteoblast-derived MVs biomimetics, as a means of reconstituting a phospholipid...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Alkaline phosphatase; Biomineralization; Calcification; Lipids; Pyrophosphate; ATP.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010000300003
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Solubilization of Na,K-ATPase from rabbit kidney outer medulla using only C12E8 BJMBR
Santos,H.L.; Lamas,R.P.; Ciancaglini,P..
SDS, C12E8, CHAPS or CHAPSO or a combination of two of these detergents is generally used for the solubilization of Na,K-ATPase and other ATPases. Our method using only C12E8 has the advantage of considerable reduction of the time for enzyme purification, with rapid solubilization and purification in a single chromatographic step. Na,K-ATPase-rich membrane fragments of rabbit kidney outer medulla were obtained without adding SDS. Optimum conditions for solubilization were obtained at 4ºC after rapid mixing of 1 mg of membrane Na,K-ATPase with 1 mg of C12E8/ml, yielding 98% recovery of the activity. The solubilized enzyme was purified by gel filtration on a Sepharose 6B column at 4ºC. Non-denaturing PAGE revealed a single protein band with...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Na; K-ATPase Rabbit kidney medulla Membrane solubilization C12E8; (alphaß)2 Dimer.
Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2002000300002
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Toluene permeabilization differentially affects F- and P-type ATPase activities present in the plasma membrane of Streptococcus mutans BJMBR
Thedei Jr.,G.; Leitão,D.P.S.; Bolean,M.; Paulino,T.P.; Spadaro,A.C.C.; Ciancaglini,P..
Streptococcus mutans membrane-bound P- and F-type ATPases are responsible for H+ extrusion from the cytoplasm thus keeping intracellular pH appropriate for cell metabolism. Toluene-permeabilized bacterial cells have long been used to study total membrane-bound ATPase activity, and to compare the properties of ATPase in situ with those in membrane-rich fractions. The aim of the present research was to determine if toluene permeabilization can significantly modify the activity of membrane-bound ATPase of both F-type and P-type. ATPase activity was assayed discontinuously by measuring phosphate release from ATP as substrate. Treatment of S. mutans membrane fractions with toluene reduced total ATPase activity by approximately 80% and did not allow...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cell permeabilization; F-type ATPase; P-type ATPase; Toluene permeabilization; Streptococcus mutans.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2008001200002
Registros recuperados: 4
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