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A continuous fluorescent assay for the determination of plasma and tissue angiotensin I-converting enzyme activity BJMBR
Alves,M.F.; Araujo,M.C.; Juliano,M.A.; Oliveira,E.M.; Krieger,J.E.; Casarini,D.E.; Juliano,L.; Carmona,A.K..
A continuous assay using internally quenched fluorescent peptides with the general sequence Abz-peptidyl-(Dnp)P-OH (Abz = ortho-aminobenzoic acid; Dnp = 2,4-dinitrophenyl) was optimized for the measurement of angiotensin I-converting enzyme (ACE) in human plasma and rat tissues. Abz-FRK(Dnp)P-OH, which was cleaved at the Arg-Lys bond by ACE, was used for the enzyme evaluation in human plasma. Enzymatic activity was monitored by continuous recording of the fluorescence (lambdaex = 320 nm and lambdaem = 420 nm) at 37ºC, in 0.1 M Tris-HCl buffer, pH 7.0, with 50 mM NaCl and 10 µM ZnCl2. The assays can be performed directly in the cuvette of the fluorimeter and the hydrolysis followed for 5 to 10 min. ACE measurements in the plasma of 80 healthy patients with...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Angiotensin-converting enzyme activity; Fluorometric assay; Rat tissue angiotensin- converting enzyme; Human plasma angiotensin-converting enzyme.
Ano: 2005 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2005000600007
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Detection of the basement membrane-degrading proteolytic activity of Paracoccidioides brasiliensis after SDS-PAGE using agarose overlays containing Abz-MKALTLQ-EDDnp BJMBR
Puccia,R.; Juliano,M.A.; Juliano,L.; Travassos,L.R.; Carmona,A.K..
We have characterized, in the Paracoccidioides brasiliensis yeast phase, an exocellular SH-dependent serine proteinase activity against Abz-MKRLTL-EDDnp and analogous fluorescent-quenched peptides, and showed that it is also active against constituents of the basement membrane in vitro. In the present study, we separated the components of P. brasiliensis culture filtrates by electrophoresis and demonstrated that the serine-thiol exocellular proteinase has a diffuse and heterogeneous migration by SDS-PAGE, localizing in a region between 69 and 43 kDa. The hydrolytic activity was demonstrable after SDS-PAGE using buffered agarose overlays of Abz-MKALTLQ-EDDnp, following incubation at 37oC, and detection of fluorescent bands with a UV transilluminator....
Tipo: Info:eu-repo/semantics/article Palavras-chave: P. brasiliensis; Serine-thiol proteinase; SDS-PAGE; Fluorescent-quenched peptides.
Ano: 1999 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1999000500019
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Glycosaminoglycans affect the interaction of human plasma kallikrein with plasminogen, factor XII and inhibitors BJMBR
Gozzo,A.J.; Nunes,V.A.; Nader,H.B.; Dietrich,C.P.; Carmona,A.K.; Sampaio,M.U.; Sampaio,C.A.M.; Araújo,M.S..
Human plasma kallikrein, a serine proteinase, plays a key role in intrinsic blood clotting, in the kallikrein-kinin system, and in fibrinolysis. The proteolytic enzymes involved in these processes are usually controlled by specific inhibitors and may be influenced by several factors including glycosaminoglycans, as recently demonstrated by our group. The aim of the present study was to investigate the effect of glycosaminoglycans (30 to 250 µg/ml) on kallikrein activity on plasminogen and factor XII and on the inhibition of kallikrein by the plasma proteins C1-inhibitor and antithrombin. Almost all available glycosaminoglycans (heparin, heparan sulfate, bovine and tuna dermatan sulfate, chondroitin 4- and 6-sulfates) reduced (1.2 to 3.0 times) the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Glycosaminoglycans; Human plasma kallikrein; Plasminogen; Factor XII; C1-inhibitor; Antithrombin.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003000800011
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The chronic blockade of angiotensin I-converting enzyme eliminates the sex differences of serum cytokine levels of spontaneously hypertensive rats BJMBR
Dalpiaz,P.L.M.; Lamas,A.Z.; Caliman,I.F.; Medeiros,A.R.S.; Abreu,G.R.; Moysés,M.R.; Andrade,T.U.; Alves,M.F.; Carmona,A.K.; Bissoli,N.S..
Sex hormones modulate the action of both cytokines and the renin-angiotensin system. However, the effects of angiotensin I-converting enzyme (ACE) on the proinflammatory and anti-inflammatory cytokine levels in male and female spontaneously hypertensive rats (SHR) are unclear. We determined the relationship between ACE activity, cytokine levels and sex differences in SHR. Female (F) and male (M) SHR were divided into 4 experimental groups each (n = 7): sham + vehicle (SV), sham + enalapril (10 mg/kg body weight by gavage), castrated + vehicle, and castrated + enalapril. Treatment began 21 days after castration and continued for 30 days. Serum cytokine levels (ELISA) and ACE activity (fluorimetry) were measured. Male rats exhibited a higher serum ACE...
Tipo: Info:eu-repo/semantics/article Palavras-chave: SHR Enalapril Sexual dimorphism Angiotensin I-converting enzyme Cytokines IL-10; IL-6 and TNF-α.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2013000200171
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