Heparin modulates the endopeptidase activity of Leishmania mexicana cysteine protease cathepsin L-Like rCPB2.8

Cysteine protease B is considered crucial for the survival and infectivity of the Leishmania in its human host. Several microorganism pathogens bind to the heparin-like glycosaminoglycans chains of proteoglycans at host-cell surface to promote their attachment and internalization. Here, we have investigated the influence of heparin upon Leishmania mexicana cysteine protease rCPB2.8 activity. The data analysis revealed that the presence of heparin affects all steps of the enzyme reaction: (i) it decreases 3.5-fold the k1 and 4.0-fold the k−1, (ii) it affects the acyl-enzyme accumulation with pronounced decrease in k2 (2.7-fold), and also decrease in k3 (3.5-fold). The large values of ΔG = 12 kJ/mol for the association and dissociation steps indicate substantial structural strains linked to the formation/dissociation of the ES complex in the presence of heparin, which underscore a conformational change that prevents the diffusion of substrate in the rCPB2.8 active site. Binding to heparin also significantly decreases the α-helix content of the rCPB2.8 and perturbs the intrinsic fluorescence emission of the enzyme. The data strongly suggest that heparin is altering the ionization of catalytic (Cys25)-S−/(His163)-Im+ H ion pair of the rCPB2.8. Moreover, the interaction of heparin with the N-terminal pro-region of rCPB2.8 significantly decreased its inhibitory activity against the mature enzyme. Taken together, depending on their concentration, heparin-like glycosaminoglycans can either stimulate or antagonize the activity of cysteine protease B enzymes during parasite infection, suggesting that this glycoconjugate can anchor parasite cysteine protease at host cell surface

Reproductive cycle and embryonic development of the gastropod Melampus coffeus (Linnaeus, 1758) (Ellobiidae) in the Brazilian Northeast

Melampus coffeus belongs to a primitive group of pulmonate mollusks found mainly in the upper levels of the marine intertidal zone. They are common in the neotropical mangroves. Little is known about the biology of this species, particularly about its reproduction. The aim of this study was to 1) characterize the morphology and histology of M. coffeus' gonad; 2) describe the main gametogenesis events and link them to a range of maturity stages; 3) chronologically evaluate the frequency of the different maturity stages and their relation to environmental factors such as water, air and sediment temperatures, relative humidity, salinity and rainfall; and 4) characterize M. coffeus' spawning, eggs and newly hatched veliger larvae. Samples were collected monthly between February, 2007 and January, 2009 from the mangroves of Praia de Arpoeiras, Acaraú County, State of Ceará, northeastern Brazil. The characterization of the gonad development stages was carried out using routine histological techniques. The results of this study show that Melampus coffeus is a simultaneous hermaphrodite. The follicles have masculine and feminine elements, interleaved within the gonad. M. coffeus presents a well-defined synchronous reproductive cycle, showing successive maturation, release and resting periods. The average diameter of the oocytes was negatively correlated with salinity and positively correlated with rainfall. The results show that no reproductive activity occurs during periods of drought. After the dry season, the increasing rainfall levels and reduced salinity lead to the appearance of very dense populations, predominantly composed of small individuals

Kuluttajabarometri maakunnittain 2000, 2. neljännes

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