Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites

One of the major questions concerning Giardia is the understanding of pathophysiological processes associated with small intestine abnormalities. There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in the host intestinal epithelium. The present investigation was undertaken to examine the protease activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). E/S products from trophozoites of each strain in conditioned medium were tested with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the protein profiles, and the protease activity was analyzed using substrate-impregnated SDS-PAGE (gelatin and collagen) and hemoglobin assay. The proteases characterization was based on inhibition assays including synthetic inhibitors. Electrophoresis analysis of E/S products revealed a banding pattern composed by few bands (4 to 6 bands) in the migration region of 123 to 28 kDa. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted substrate degradation and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibitor assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases, although the presence of serine proteases was also indicated. Degradation of substrates including collagen and hemoglobin could lead us to speculate different functions of Giardia excreted/secreted proteases in vivo, but to confirm this possibility and to elucidate its implication on host-parasite interactions, further experiments applying protocols for the purification of proteases are necessary. Even so, our observations are relevant and hold the perspective for the understanding about protease activity in Giardia trophozoites of axenic strain isolated in an endemic area.

Characterization of the excretory/secretory products of Dermatobia hominis larvae, the human bot fly

Proteolytic activity in excretory/secretory products (ESP) of first- (L1), second- (L2) and third-instar (L3) larvae of Dermatobia hominis was analyzed through gelatin-gel and colorimetric enzyme assays with the chromogenic substrates azocasein and BApNA. The functional characterization of proteases was based on inhibition assays including synthetic inhibitors. ESP were obtained from new-hatched larvae reared in the laboratory and from second- and third-instar larvae removed from naturally infested cattle. Gelatin-gel analysis evidenced few bands of proteolysis, predominantly of high apparent molecular masses, in ESP of L1, whereas in the gel of L2 and U ESP there was a wide range of proteolytic activity most of them not resolved in a single species. Azocasein assays revealed a progressive increase of protease activity from first- to third-instar larvae. Protease inhibitor assays revealed a predominance of metalloproteases in L1 ESP that could be related to a skin penetration process and to a diversion of host immune response. The predominance of serine proteases in L2 and L3 and the great tryptic activity presented by L3 ESP were attributed to an increasing trophic activity by the growing larvae, since the viability of adult flies strictly depends on larval abilities to assimilate nutrients from the host. Taking together, these results suggest that Dematobia larvae secrete/excrete different proteases that may be related to diverse functions during host penetration and infestation, which reinforces the relevance of the study of such proteolytic enzymes. (C) 2009 Elsevier B.V. All rights reserved.

Interaction between Advanced Glycation End Products Formation and Vascular Responses in Femoral and Coronary Arteries from Exercised Diabetic Rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Pectinase production by Penicillium viridicatum RFC3 by solid state fermentation using agricultural wastes and agro-industrial by-products

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Sensory acceptability of raw and extruded bovine rumen protein in processed meat products

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Friction losses in valves and fittings for liquid food products

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

On the calculation of inner products of Schur functions

Two methods for calculating inner products of Schur functions in terms of outer products and plethysms are given and they are easy to implement on a machine. One of these is derived from a recent analysis of the SO(8) proton-neutron pairing model of atomic nuclei. The two methods allow for generation of inner products for the Schur functions of degree up to 20 and even beyond.

Electrochemical Reduction as a Powerful Tool to Highlight the Possible Formation of By-Products More Toxic Than Sudan III Dye

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Development of a sensitive potentiometric sensor for determination of fumaric acid in powdered food products

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Inactivation and disposal of by-products from Mycobacterium smegmatis by photoelectrocatalytic oxidation using Ti/TiO2-Ag nanotube electrodes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Assessment of by-products of chlorination and photoelectrocatalytic chlorination of an azo dye

The present work describes a more efficient methodology for the chlorination of water containing disperse dyes, where the chlorinated byproducts identified by mass spectra are compared. this investigation, we tested the degradation of Cl Disperse Blue 291 dye, 2-[(2-Bromo-4,6-dinitrophenyl)azo]-5-(diethylamino)-4-methoxyacetanilide) a commercial azo dye with mutagenic properties. The present work evaluates the photoelectrocatalytic efficiency of removing the Cl Disperse Blue 291 dye from a wastewater of the textile industry. We employed NaCl as a supporting electrolyte. It should be noted that photoelectrocatalytic techniques are non-conventional method of generating chlorine radicals. The by-products formed in this process were analyzed using spectrophotometry, liquid chromatography, dissolved organic carbon, mass spectral analysis and mutagenicity assays. The process efficiency was compared with the conventional chlorination process adopted during sewage and effluents treatment processes. This conventional chlorination process is less efficient in removing color, total organic carbon than the photoelectrochemistry technique. Furthermore, we shall demonstrate that the mutagenicity of the generated by-products obtained using photoelectrocatalysis is completely different from that obtained by the conventional oxidation of chloride ions in the drinking wafer treatment process. (C) 2012 Published by Elsevier B.V.

Determination of nitrite in meat products and water using dapsone with combined spot test/diffuse reflectance on filter paper

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Analyses of the genotoxic and mutagenic potential of the products formed after the biotransformation of the azo dye Disperse Red 1

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)