Biomedical subjects

L C B B Coelho

Publications and source records attributed to L C B B Coelho.

10 recordsLinked to original sources

PubMed2006

Purification and primary structure determination of two Bowman-Birk type trypsin isoinhibitors from Cratylia mollis seeds.

Two Bowman-Birk type trypsin inhibitors (CmTI(1) and CmTI(2)) were purified from Cratylia mollis seeds by acetone precipitation, ion exchange, gel filtration and reverse-phase chromatography. CmTI(1) and CmTI(2), with 77 and 78 amino acid residues, respectively, were sequenced in their entirety and show a high structural similarity to Bowman-Birk inhibitors from other Leguminosae. The putative reactive sites of CmTI(1) are a lysine residue at position 22 and a tyrosine residue at position 49. Different reactive sites, as identified by their alignment with related inhibitors, were found for CmTI(2): lysine at position 22 and leucine at position 49. The dissociation constant K(i) of the complex with trypsin is 1.4 nM. The apparent molecular mass is 17 kDa without DDT and 11 kDa with reducing agent and heating.

Amino Acid Sequence↗

PubMed2005

Yet another application of the Monte Carlo method for modeling in the field of biomedicine.

By means of Monte Carlo simulations performed in the C programming language, an example of scientific programming for the generation of pseudorandom numbers relevant to both teaching and research in the field of biomedicine is presented. The relatively simple algorithm proposed makes possible the statistical analysis of sequences of random numbers. The following three generators of pseudorandom numbers were used: the rand function contained in the stdlib.h library of the C programming language, Marsaglia's generator, and a chaotic function. The statistical properties of the sequences generated were compared, identical parameter values being adopted for this purpose. The properties of two estimators in finite samples of the pseudorandom numbers were also evaluated and, under suitable conditions, both the maximum-likelihood and method of moments proved to be good estimators. The findings demonstrated that the proposed algorithm appears to be suitable for the analysis of data from random experiments, indicating that it has a large variety of possible applications in the clinical practice.

Algorithms↗

PubMed2005

Detection of water soluble lectin and antioxidant component from Moringa oleifera seeds.

Seed flour from Moringa oleifera is widely used as a natural coagulant for water treatment in developing countries. Extracts obtained by water soaking of M. oleifera intact seeds were investigated for the presence of lectin, trypsin inhibitor, tannin as well as antioxidant activity. A water soluble M. oleifera lectin (WSMoL) detected was mainly active with rabbit cells at pH 4.5; heat treatment, pH 7.0, fructose and porcine thyroglobulin abolished HA of WSMoL. Trypsin inhibitor or tannins were not detected; the antioxidant component (WSMoAC) reduced 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) was slower than catechin and was thermostable. The extracts showed a primary glycopolypeptide band of Mw 20,000; the main native acidic protein showed hemagglutinating activity. WSMoL may be involved in seed coagulant properties.

Animals↗

PubMed2004

Antioxidant activity of leaf extracts from Bauhinia monandra.

Bauhinia monandra Kurz. is used in Brazil for the treatment of diabetes. Since this activity may be correlated with the presence of antioxidant compounds, leaf extracts of B. monandra were evaluated for their radical scavenging capacity (RSC). An ethanolic extract was taken up in aqueous methanol and partitioned with hexane, chloroform, ethyl acetate to yield three organic extracts together with remaining aqueous extract. The RSC was determined spectrophotometrically using 1,1-diphenylpicrylhydrazyl free radical (DPPH). The chloroform and ethyl acetate extracts were the most appropriate as sources of antioxidant compounds as shown by their inhibition concentration (IC50) and inhibition percentage (IP) values. The antioxidant activity of such extracts was attributed to the presence of three compounds of different polarities (flavonoids and steroids). The chloroform and ethyl acetate extracts exhibited an IC50 of approximately 2 mg/g DPPH and IP values in the range of 60-65%. The results indicate that the extracts of B. monandra have a very potent antioxidant activity, compared with the pure catechins used as positive controls and with other plant extracts.

Acetates↗

PubMed2004

Ultrastructural analysis and immunocytochemical localization of isolectins in Cratylia mollis seeds.

Cratylia mollis is a native forage from the semi-arid region of Northeast, State of Pernambuco, Brazil, whose seeds have been considered an important lectin source. Multiple molecular forms of lectins, carbohydrate-binding proteins, have been purified from C. mollis seeds (Cra Iso) allowing several applications of these purified proteins. In this work seeds were processed for ultrastructural analysis and immunocytochemical localization of the two most abundant isolectins, Cra Iso 1 and Cra Iso 3, with glucose/mannose and galactose specificities, respectively. The ultrastructural analysis revealed a typical plant cell: organelles, nucleus and cellular wall were visualized. The localization of isolectins occurred mainly in the amorphous matrix of protein bodies, and in the cellular walls of the embryonic axis. The results showed that the isolectins, which differ in relation to carbohydrate specificity and glycosylation are located in the same cellular compartment suggesting different functions inside the same subcellular organelle. Cra Iso 1 and Cra Iso 3 distribution in the C. mollis seeds was consistent with the subcellular localization of several legume lectins.

Animals↗

PubMed2003

Novel core(polyester)-shell(polysaccharide) nanoparticles: protein loading and surface modification with lectins.

This study describes new lectin-decorated or protein-loaded nanoparticles with a hydrophobic poly(epsilon-caprolactone) (PCL) core and a hydrophilic dextran (Dex) corona. In this view, a family of block Dex-PCLn copolymers was first synthesized, consisting of a Dex backbone to which n preformed PCL blocks were grafted. The ability of these new copolymers to form nanoparticles was evaluated in comparison with a series of PCL homopolymers of various molecular weights (2000, 10,000 and 40,000 g/mole). Two different nanoparticle preparation methods have been developed and tested for their efficacy to incorporate proteins. For this, three proteins were used: a model protein, bovine serum albumin (BSA), a lectin from leaves of Bauhinia monandra (BmoLL) and Lens culinaris (LC) lectin. All these proteins were successfully incorporated in nanoparticles with a mean diameter around 200 nm. Lectins could also be adsorbed onto the surface of Dex-PCLn nanoparticles. Surface-bound BmoLL conserved its hemagglutinating activity, suggesting the possible application of this type of surface-modified nanoparticles for targeted oral administration. Caco-2 cellular viability was higher than 70% when put in contact with Dex-PCLn nanoparticles, even at concentrations as high as 660 microg/ml.

Animals↗

PubMed2003

Isolation of a trypsin inhibitor from Echinodorus paniculatus seeds by affinity chromatography on immobilized Cratylia mollis isolectins.

A highly purified trypsin inhibitor was obtained from Echinodorus paniculatus when an extract prepared from E. paniculatus seed flour (25 gl(-1), with 0.1 M ammonium acetate buffer, pH 8.3, under agitation for 6 min at 28 degrees C) was chromatographed on Sephadex G-25 (12 mlh(-1)), followed by affinity chromatography on immobilized Cratylia mollis isolectins (Cra Iso 1,2,3-Sepharose). The column chromatography was performed at 24 degrees C; the matrix was washed (30 mlh(-1)) with 0.1 M sodium phosphate buffer, pH 7.4 or with the same buffer containing 0.2 M glucose, followed by application of inhibitor sample and elution with 0.015 M sodium borate buffer, pH 7.4, or 1.0 M NaCl. A purified fraction of inhibitor was obtained by gel filtration chromatography (GF-450/HPLC column). Trypsin inhibitory activity was eliminated when the inhibitor was treated with metaperiodate showing that the carbohydrate moiety was important for trypsin inhibition. Binding of inhibitor was also evaluated on immobilized concanavalin A (Con A-Sepharose) using previously described chromatographic conditions with results similar to Cra Iso 1,2,3-Sepharose chromatography.

Alismataceae↗

PubMed2003

Electrochemical potential of free and immobilized Cratylia mollis seed lectin.

The electrochemical potentials for free or immobilized Cratylia mollis seed lectin (Cra) were obtained through potentiostatic techniques. A saline solution was used as support to control the charge distribution between saturated calomel electrode and platinum electrode (working electrode). The electrochemical potential to free Cra was determined at the following concentrations: 0.6, 0.9 and 1.0 mg/ml in an aerated environment under different temperatures (5, 10 and 20 degrees C). The best electrochemical potential was obtained with 1.0 mg/ml, at 5 and 10 degrees C, 87 and 102 mV, respectively. Electrochemical potential to Cra immobilized on glass beads activated with 3-aminopropyltriethoxysilane described a linear behavior in relation to the increase in glucose concentration. The development of techniques to define interface electrical parameters will be able to give information about charged groups adsorbed to electrode surface revealing interactions particularly in biological systems.

Electrochemistry↗

PubMed2003

Parkia pendula lectin as histochemistry marker for meningothelial tumour.

Lectins have been intensively used in histochemical techniques for cell surface characterization. These proteins are involved in several biological processes and their use as histochemical markers have been evaluated since they can indicate differences in cell surfaces. Parkia pendula lectin (PpeL) was evaluated as histochemical marker for meningothelial meningioma biopsies. Tissue slices were incubated with PpeL conjugated to horseradish peroxidase (PpeL-HRP) and Concanavalin A-HRP (ConA-HPR) and the binding visualized with diaminobenzidine and hydrogen peroxide. The lectin-tissue binding was inhibited with D-glucose. PpeL showed to be a useful tool for the characterization of meningothelial tumour and clinico-pathological diagnosis.

Adult↗

PubMed2002

Production and characterization of a thermostable glucoamylase from Streptosporangium sp. endophyte of maize leaves.

Thermostable amylolytic enzymes are currently investigated to improve industrial processes of starch degradation. Streptosporangium sp. an endophytic actinomycete isolated from leaves of maize (Zea mays L.) showed glucoamylase production, using starch-Czapek medium, and the highest rate was obtained in the initial growth phase, after incubation for 24 h at pH 8.0. Maximum glucoamylase activity (158 U mg(-1) protein) was obtained at pH 4.5 and 70 degrees C. The isolated enzyme exhibited thermostable properties as indicated by retention of 100% of residual activity at 70 degrees C for 30 min with total inhibition at 100 degrees C. Extracellular enzyme from Streptosporangium sp. was purified by fractionated precipitation with ammonium sulphate. After 60% saturation produced 421 U mg(-1) protein, and yield was 74% with purification 2.7 fold. The enzyme produced by Streptosporangium sp. has potential for industrial applications.

Actinomycetales↗