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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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Association of urinary 90 kDa angiotensin- converting enzyme with family history of hypertension and endothelial function in normotensive individuals BJMBR
Teixeira,A.M.S.; Plavnik,F.L.; Fernandes,F.B.; Marson,O.; Christofalo,D.M.J.; Ajzen,S.A.; Sesso,R.; Franco,M.C.; Casarini,D.E..
We described angiotensin-I-converting enzyme (ACE) isoforms with molecular masses of 190, 90, and 65 kDa in the urine of normotensive offspring of hypertensive subjects. Since they did not appear in equal amounts, we suggested that 90 kDa ACE might be a marker for hypertension. We evaluated the endothelial response in normotensive offspring with or without family history of hypertension and its association with the 90 kDa ACE in urine. Thirty-five normotensive subjects with a known family history of hypertension and 20 subjects without a family history of hypertension, matched for age, sex, body weight, and blood pressure, were included in the study. Endothelial function was assessed by ultrasound and a sample of urine was collected for determination of...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Endothelial dysfunction; Angiotensin-I-converting enzyme; Hypertension; Normotension; N-domain angiotensin- converting enzyme.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2008000500002
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Characterization of renin mRNA expression and enzyme activity in rat and mouse mesangial cells BJMBR
Andrade,A.Q.; Casarini,D.E.; Schor,N.; Boim,M.A..
Renin is an enzyme involved in the stepwise generation of angiotensin II. Juxtaglomerular cells are the main source of plasma renin, but renin activity has been detected in other cell types. In the present study we evaluated the presence of renin mRNA in adult male Wistar rat and mouse (C-57 Black/6) mesangial cells (MC) and their ability to process, store and release both the active and inactive forms of the enzyme. Active renin and total renin content obtained after trypsin treatment were estimated by angiotensinogen consumption analyzed by SDS-PAGE electrophoresis and quantified by angiotensin I generation by HPLC. Renin mRNA, detected by RT-PCR, was present in both rat and mouse MC under basal conditions. Active renin was significantly higher...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Renin; Prorenin; Mesangial cell; Kidney.
Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2002000100003
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High expression of human carboxypeptidase M in Pichia pastoris: Purification and partial characterization BJMBR
Craveiro,R.B.; Ramalho,J.D.; Chagas,J.R.; Wang,P.H.M.; Casarini,D.E.; Pesquero,J.L.; Araújo,R.C.; Pesquero,J.B..
Carboxypeptidase M (CPM) is an extracellular glycosylphosphatidyl-inositol-anchored membrane glycoprotein, which removes the C-terminal basic residues, lysine and arginine, from peptides and proteins at neutral pH. CPM plays an important role in the control of peptide hormones and growth factor activity on the cell surface. The present study was carried out to clone and express human CPM in the yeast Pichia pastoris in order to evaluate the importance of this enzyme in physiological and pathological processes. The cDNA for the enzyme was amplified from total placental RNA by RT-PCR and cloned in the vector pPIC9, which uses the methanol oxidase promoter and drives the expression of high levels of heterologous proteins in P. pastoris. The cpm gene, after...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Kinins; Human carboxypeptidase M; Recombinant protein; Pichia pastoris.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000200007
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Relationship of cyclosporin and sirolimus blood concentrations regarding the incidence and severity of hyperlipidemia after kidney transplantation BJMBR
Spinelli,G.A.; Felipe,C.R.; Machado,P.G.; Garcia,R.; Casarini,D.E.; Moreira,S.R.; Park,S.I.; Tedesco-Silva Jr.,H.; Medina-Pestana,J.O..
The influence of drug concentrations on the development of persistent posttransplant hyperlipidemia was investigated in 82 patients who received cyclosporin A (CsA) and prednisone plus sirolimus (SRL) (52) or azathioprine (AZA) (30) during the first year after transplantation. Blood levels of CsA and SRL, daily doses of AZA and prednisone, and cholesterol, triglyceride, and glucose concentrations were determined during each visit (pretransplant and 30, 60, 90, 120, 180, and 360 days posttransplant). Persistent hyperlipidemia was defined as one-year average steady-state cholesterol (CavCHOL) or triglyceride (CavTG) concentrations above 240 and 200 mg/dL, respectively. Mean cholesterol and triglyceride concentrations increased after transplantation (P <...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Hyperlipidemia; Cholesterol; Triglyceride; Kidney transplantation; Cyclosporin A; Sirolimus.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000100003
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Swimming training increases cardiac vagal activity and induces cardiac hypertrophy in rats BJMBR
Medeiros,A.; Oliveira,E.M.; Gianolla,R.; Casarini,D.E.; Negrão,C.E.; Brum,P.C..
The effect of swimming training (ST) on vagal and sympathetic cardiac effects was investigated in sedentary (S, N = 12) and trained (T, N = 12) male Wistar rats (200-220 g). ST consisted of 60-min swimming sessions 5 days/week for 8 weeks, with a 5% body weight load attached to the tail. The effect of the autonomic nervous system in generating training-induced resting bradycardia (RB) was examined indirectly after cardiac muscarinic and adrenergic receptor blockade. Cardiac hypertrophy was evaluated by cardiac weight and myocyte morphometry. Plasma catecholamine concentrations and citrate synthase activity in soleus muscle were also determined in both groups. Resting heart rate was significantly reduced in T rats (355 ± 16 vs 330 ± 20 bpm). RB was...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Swimming-training exercise; Cardiac autonomic balance; Hemodynamics; Hypertrophic effect; Parasympathetic system; Resting bradycardia.
Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2004001200018
Registros recuperados: 6
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