Characterization of a bovine collagen-hydroxyapatite composite scaffold for bone tissue engineering.

Different biomaterials have been used as scaffolds for bone tissue engineering. Here we characterize a biomaterial composed of sintered (1100 degrees C) and powdered hydroxyapatite (HA) and type I collagen (Coll), both of bovine origin, designed for osteoconductive and osteoinductive scaffolds. Coll/HA proportions were 1/2.6 and 1/1 (wet weight), and particles sizes varied from 200 to 400 microm. Vv (volume density) and Sv (surface to volume density) for the HA particles in the composite ranged from 0.48 +/- 0.06 to 0.55 +/- 0.02 and 5.090 +/- 0.545 to 6.366 +/- 0.289 microm(-1), respectively. Due to the relatively small changes in Vv and Sv, a macroporosity could be characterized for the biocomposite. X-ray diffraction and infrared spectroscopy showed that the sintered bone was composed essentially of HA with minimum additional groups such as surface calcium hydroxide, surface and crystal water, free carbon dioxide and possibly brushite. Mass spectrometry detected carbonates at A and B sites of HA, and weakly bound to the structure. Human osteoblasts adhered and spread on both the HA particle surface and the collagen fibers, which seemed to guide cells between adjacent particles. The biocomposite studied has several characteristics considered as ideal for its use as a scaffold for osteoconduction and osteoinduction.

Diagnosis of dengue infection by detecting specific immunoglobulin M antibodies in saliva samples.

To investigate whether saliva could be used for diagnosis of recent dengue, serum and saliva samples were collected simultaneously from patients with suspected dengue infection. Sera (1:10 dilution) and saliva (undiluted) were tested by using an IgM capture enzyme linked immunosorbent assay (MAC-ELISA) with minor modifications (serum and saliva absorption for 3 h at 37 degrees C). The quality of saliva was evaluated by determining the IgG total concentration (enzyme immunoassay) which ranged from 2.7 to > 50 mg/l. Recent dengue infection was confirmed in 38 cases. Forty-six serum and saliva specimens were collected from these patients 1-30 days after the onset of symptoms. IgM was detected in 65.8% saliva samples. High rate of positivity ( > 80%) was observed for the saliva samples collected > or = 5 days after the onset of the disease. Fifty serum and saliva samples from other 32 patients with rash diseases were also tested and all the specimens were unreactive by MAC-ELISA. These results indicate that saliva may be a convenient non-invasive alternative to serum for diagnosis of recent dengue fever infection, especially for epidemiological studies during outbreaks of the disease.

[Multicenter study of intracoronary stent implantation in Rio de Janeiro]. / Estudo multicêntrico do implante de stent intracoronário no Rio de Janeiro.

PURPOSE: To evaluate retrospectively stent implantation (SI) in patients with coronary artery disease (CAD) performed in 7 hospitals in Rio de Janeiro. METHODS: From June/94 to December/96 2,220 procedures were performed among which we analyzed 783 SI in 660 (29.7%) patients using coronary angiography without digital subtraction. Several types of stents were used: Palmaz-Schatz (40.9%), Gianturco-Roubin (29.1%) e NIR (22.0%). Indications for SI: 1--de novo lesion (67.9%); 2) restenotic lesion (16.0%); 3) sub-optimal results after PTCA (8.2%); 4) abrupt or threatened closure after PTCA (4.9%); 5) chronic occlusion (3.0%). All stents were implanted using high pressure balloon inflation without intracoronary ultrasound guidance. Sub-acute stent thrombosis prevention, in the majority of patients (87.8%) was done with ticlopidine and aspirin. RESULTS: Early outcome: a) the success rate of SI in 770 lesions was 98.0% in 646 (97.9%) patients; b) the clinical success rate in 634 patients was 96.0%; c) the major complications were acute myocardial infarction (1.1%); coronary artery bypass graft (1.4%) and death (0.8%); d) vascular complications with surgical correction and/or bleeding occurred in 3.0%. Late outcome: a) the clinical follow-up of 399 (60.4%) and the coronary angiographies of 121 patients (30.3%) showed in-stent lesion in 79 (19.8%); b) other event rates: myocardial infarction (1.5%); coronary bypass (2.3%); death: 1.0% and other PTCA or similar procedure (12.5%). CONCLUSION: This multicentric study showed that SI for CAD can be performed with safety, high early success rate, few complications and low rate of cardiac events during the late follow-up.

[Stents for bifurcational coronary lesion]. / Implante de stent em bifurcação coronária.

We describe two cases of coronary angioplasty with stents implantation in bifurcational lesion of the left anterior descending artery and the diagonal branch using new techniques successfully performed. Angiographic reestenosis was not present after seven months in one case and the patient was asymptomatic after six months of clinical follow-up in the other.

Kinetic characterization of rat tissue kallikrein using N alpha-substituted arginine 4-nitroanilides and N alpha-benzoyl-L-arginine ethyl ester as substrates.

Hydrolysis of seven N alpha-substituted L-arginine 4-nitroanilides: benzoyl-arginine p-nitroanilide (Bz-Arg-Nan), tosyl-arginine p-nitroanilide (Tos-Arg-Nan), acetyl-leucyl-arginine p-nitroanilide (Ac-Leu-Arg-Nan), acetyl-phenylalanyl-arginine p-nitroanilide (Ac-Phe-Arg-Nan), benzoyl-phenylalanyl-arginine p-nitroanilide (Bz-Phe-Arg-Nan), tosyl-phenylalanyl-arginine p-nitroanilide (Tos-Phe-Arg-Nan), and D-valyl-leucyl-arginine p-nitroanilide (D-Val-Leu-Arg-Nan), and the N alpha-substituted L-arginine ester: benzoyl-arginine ethyl ester (Bz-Arg-OEt), by rat tissue kallikrein was studied throughout a wide range of substrate concentrations. The enzyme showed a bimodal behavior with all the substrates tested except Tos-Arg-Nan. At low substrate concentrations (10 to 170 microM for p-nitroanilides and 50 to 190 microM for Bz-Arg-OEt) the hydrolysis followed Michaelis-Menten kinetics, but at higher substrate concentrations (150 to 700 microM for p-nitroanilides and 200 to 1800 microM for Bz-Arg-OEt) a deviation from Michaelis-Menten kinetics was observed with a significant decrease in hydrolysis rates. At high concentrations of the p-nitroanilide substrates, partial enzyme inhibition was observed, whereas complete enzyme inhibition was observed with Bz-Arg-OEt at high concentration. The kinetic parameters reported here were calculated from data for substrate concentrations range where the enzyme followed Michaelis-Menten behavior. D-Val-Leu-Arg-Nan (Km = 24 +/- 2 microM; Vmax = 10.42 +/- 0.28 microM/min) was the best substrate tested, followed by Ac-Phe-Arg-Nan (Km = 13 +/- 2 microM; Vmax = 3.21 +/- 0.11 microM/min), while Tos-Arg-Nan (Km = 29 +/- 2 microM; Vmax = 0.10 +/- 0.002 microM/min) was the worst of the tested substrates for rat tissue kallikrein. For the hydrolysis of Bz-Arg-OEt (Km = 125 +/- 15 microM; Vmax = 121.3 +/- 7.6 microM/min), the kinetic parameters using a substrate inhibition model can reasonably account for the observed enzyme behavior, with a Ksi value about 13.6 times larger than the estimated Km value.

Kinetic characterization of rat tissue kallikrein using NÓ-substituted arginine 4-nitroanilides and NÓ-benzoyl-L-arginine ethyl ester as substrates

Hydrolysis of seven N(alpha-substituted L-arginine 4-nitroanilides: henzoyl-arginine p-nitroanilide (Bz-Arg-Nan), tosyl-arginine p-nitroanilide (Tos-Arg-Nan), acetyl-leucyl-arginine p-nitroanilide (Ac-Leu-Arg-Nan), acetyl-phenylalanyl-arginine p-nitroanilide (Ac-Phe-Arg-Nan), benzoyl-phenylalanyl-arginine p-nitroanilide (Bz-Phe-Arg-Nan), tosyl-phenylalanyl-arginine p-nitroanilide (Tos-Phe-Arg-Nan), and D-valyl-leucyl-arginine p-nitroanilide (D-Val-Leu-Arg-Nan), and the N(alpha-substituted L-arginine ester: benzoyl-arginine ethyl ester (Bz-Arg-OEt), by rat tissue kallikrein was studied throughout a wide range of substrate concentrations. The enzyme showed a bimodal behavior with all the substrates tested except Tos-Arg-Nan. At low substrate concentrations (10 to 170 muM for p-nitroanilides and 50 to 190 muM for Bz-Arg-OEt) the hydrolysis followed Michaelis-Menten kinetics, but at higher substrate concentrations (150 to 700 muM for p-nitroanilides and 200 to 1800 muM for Bz-Arg-OEt) a deviation from Michaelis-Menten kinetics was observed with a significant decrease in hydrolysis rates. At high concentrations of the p-nitroanilide substrates, partial enzyme inhibition was observed, whereas complete enzyme inhibition was observed with Bz-Arg-OEt at high concentration. The kinetic parameters reported here were calculated from data for substrate concentrations range where the enzyme followed Michaelis-Menten behavior. D-Val-Leu-Arg-Nan (Km = 24 ñ 2 muM; Vmax 10.42 ñ 0.28 muM/min) was the best substrate tested, followed by Ac-Phe-Arg-Nan (Km = 13 ñ 2 muM; Vmax = 3.21 ñ 0.11 muM/min), while Tos-Arg-Nan (Km = 29 ñ 2 muM; Vmax, = 0. 10 ñ 0.002 muM/min) was the worst of the tested substrates for rat tissue kallikrein. For the hydrolysis of Bz-Arg-OEt (Km = 125 ñ 15 muM; Vmax = 121.3 ñ 7.6 muM/min), the kinetic parameters using a substrate inhibition model can reasonably account for the observed enzyme behavior, with a Ksi value about 13.6 times larger than the estimated Km value.