Preparation and characterization of D, L-PLA loaded 17-beta-Estradiol valerate by emulsion/evaporation methods

PLA microparticles containing 17-beta-estradiol valerate were prepared by an emulsion/evaporation method in order to sustain drug release. This system was characterized concerning particle size, particle morphology and the influence of formulation and processing parameters on drug encapsulation and in vitro drug release. The biodegradation of the microparticles was observed by tissue histological analysis. Scanning electron microscopy and particle size analysis showed that the microparticles were spherical, presenting non-aggregated homogeneous surface and had diameters in the range of 718-880 nm (inert microparticles) and 3-4 mu m (drug loaded microparticles). The encapsulation efficiency was similar to 80%. Hormone released from microparticles was sustained. An in vivo degradation experiment confirmed that microparticles are biodegradable. The preparation method was shown to be suitable, since the morphological characteristics and efficiency yield were satisfactory. Thus, the method of developed microparticles seems to be a promising system for sustained release of 17-beta-estradiol.

Computer-aided Drug Design of Novel PLA(2) Inhibitor Candidates for Treatment of Snakebite

Phospholipases A(2) (PLA(2)) are enzymes commonly found in snake venoms from Viperidae and Elaphidae families, which are major components thereof. Many plants are used in traditional medicine its active agents against various effects induced by snakebite. This article presents the PLA(2) BthTX-I structure prediction based on homology modeling. In addition, we have performed virtual screening in a large database yielding a set of potential bioactive inhibitors. A flexible docking program was used to investigate the interactions between the receptor and the new ligands. We have performed molecular interaction fields (MIFs) calculations with the phospholipase model. Results confirm the important role of Lys49 for binding ligands and suggest three additional residues as well. We have proposed a theoretically nontoxic, drug-like, and potential novel BthTX-I inhibitor. These calculations have been used to guide the design of novel phospholipase inhibitors as potential lead compounds that may be optimized for future treatment of snakebite victims as well as other human diseases in which PLA(2) enzymes are involved.

Shared structural determinants for the calcium-independent liposome membrane permeabilization and sarcolemma depolarization in Bothropstoxin-I, a LYS49-PLA(2) from the venom of Bothrops jararacussu

The structural determinants of myotoxicity of bothropstoxin-I (BthTX-I), a Lys49 phospholipase A(2) from Bothrops jararacussu venom, were studied by measuring the resting membrane potential in the mouse phrenic nerve-diaphragm preparation. This method proved to be around 100-fold more sensitive than the creatine kinase release assay, and was used to evaluate a total of 31 site-directed BthTX-I alanine scanning mutants. Mutants that reduced the resting membrane potential were located in a surface patch defined by residues in the C-terminal loop (residues 115-129), positions 37-39 in the membrane interfacial recognition surface (Y46 and K54), and residue K93. These results expand the known structural determinants of the biological activity as evaluated by previous creatine kinase release experiments. Furthermore, a strong correlation is observed between the structural determinants of sarcolemma depolarization and calcium-independent disruption of liposome membranes, suggesting that a common mechanism of action underlies the permeabilization of the biological and model membranes. (C) 2009 Elsevier Ltd. All rights reserved.

The interaction of bothropstoxin-I (Lys49-PLA(2)) with liposome membranes

Bothropstoxin-I (BthTx-I) is a Lys49-PLA(2) from the venom of the snake Bothrops jararacussu, which permeabilizes biological and artificial membranes by a mechanism independent of lipid hydrolysis. This mechanism has been investigated by studying the interaction of nine single tryptophan BthTx-I mutants with negatively charged phospholipid membranes. Changes in the solvent exposure of the tryptophan in each mutant were evaluated comparing the rate of chemical modification (k(mod)) by bromosuccinamide with the maximum intrinsic tryptophan fluorescence emission wavelength (lambda(max)) in buffer and in the presence of 10% DMPA/90% DPPC liposomes. No changes in lambda(max). were observed, whereas k(mod) values for tryptophans at positions 7, 10, 31 and 125 were significantly reduced in the presence of lipids, suggesting that bound phospholipid decreases solvent accessibility at these positions. Since the half-lives of the fluorescence and chemical modification effects differ by at least six orders of magnitude, these results suggest that the bound phospholipid may interact with multiple locations on the protein surface over micro- to millisecond timescales. (C) 2009 Elsevier Ltd. All rights reserved.

Reduced pH induces an inactive non-native conformation of the monomeric bothropstoxin-I (Lys49-PLA(2))

Bothropstoxin-I (BthTx-I), a Lys49-PLA(2) from Bothrops jararacussu venom, permeabilizes membranes by a non-hydrolytic Ca(2+)-independent mechanism. The BthTx-I showed activity against liposomes including 10% and 50% negatively charged lipids at pH 7.0, but not at pH 5.0. Nevertheless, ultracentrifugation and FRET demonstrated that at pH 5.0 the BthTx-I is bound to 50% negatively charged membranes. ANS binding identified a non-native monomeric conformation at pH 5.0, suggesting that tertiary structure alterations result in activity loss of the BthTx-I at low pH. (C) 2009 Elsevier Ltd. All rights reserved.

Cognitive training increases platelet PLA(2) activity in healthy elderly subjects

Phospholipases A(2) (PLA(2)) are ubiquitous enzymes involved in membrane fatty acid metabolism and intracellular signalling. Recent studies have shown that PLA(2) subtypes are implicated in the modulation of pathways related to memory acquisition and retrieval. We investigated the effects of cognitive training on platelet PLA(2) activity in healthy elderly individuals. Twenty-three cognitively unimpaired older adults were randomly assigned to receive memory training or standard outpatient care only. Both groups were cognitively assessed by the same protocol, and the experimental group (EG) underwent a four-session memory training intervention. Pre- and post-test measures included prose and list recall, WAIS-III digit symbol, strategy use measures and platelet PLA(2) group activity. After cognitive training, patients in the EG group had significant increase in cytosolic, calcium-dependent PLA(2) (cPLA(2)), extracellular (or secreted), calcium-dependent PLA(2) (sPLA(2)), total platelet PLA(2) activity, and significant decrease in platelet calcium-independent PLA(2) (iPLA(2)) activity. Our results suggest that memory training may have a modulating effect in PLA(2)-mediated biological systems associated with cognitive functions and neurodegenerative diseases. (c) 2008 Elsevier Ltd. All rights reserved.

Neuromuscular activity of BaTX, a presynaptic basic PLA(2) isolated from Bothrops alternatus snake venom

We have previously isolated a Lys49 phospholipase A(2) homolog (BaTX) from Bothrops alternatus snake venom using a combination of molecular exclusion chromatography and reverse phase HPLC and shown its ability to cause neuromuscular blockade. In this work, we describe a one-step procedure for the purification of this toxin and provide further details of its neuromuscular activity. The toxin was purified by reverse phase HPLC and its purity and molecular mass were confirmed by SIDS-PAGE, MALDI-TOF mass spectrometry, amino acid analysis and N-terminal sequencing. BaTX (0.007-1.4 mu M) produced time-dependent, irreversible neuromuscular blockade in isolated mouse phrenic nerve-diaphragm and chick biventer cervicis preparations (time to 50% blockade with 0.35 mu M toxin: 58 +/- 4 and 24 +/- 1 min, respectively; n = 3-8; mean +/- S.E.) without significantly affecting the response to direct muscle stimulation. In chick preparations, contractures to exogenous acetylcholine (55 and 110 mu M) or KCl (13.4 mM) were unaltered after complete blockade by all toxin concentrations. These results, which strongly suggested a presynaptic mechanism of action for this toxin, were reinforced by (1) the inability of BaTX to interfere with the carbachol-induced depolarization of the resting membrane, (2) a significant decrease in the frequency and amplitude of miniature end-plate potentials, and (3) a significant reduction (59 +/- 4%, n=12) in the quantal content of the end-plate potentials after a 60 min incubation with the toxin (1.4 mu M). In addition, a decrease in the organ bath temperature from 37 degrees C to 24 degrees C and/or the replacement of calcium with strontium prevented the neuromuscular blockade, indicating a temperature-dependent effect possibly mediated by enzymatic activity. (C) 2009 Elsevier Inc. All rights reserved.

Insights on calcium-independent phospholipid membrane damage by Lys49-PLA(2) using tryptophan scanning mutagenesis of bothropstoxin-I from Bothrops jararacussu

Bothropstoxin-I (BthTx-I) is a homodimerie Lys49-PLA(2) from the venom of the snake Bothrops jararacussu, which lacks hydrolytic activity against phospholipid substrates, yet permeabilizes membranes by a Ca2+- independent mechanism. The interaction of the BthTx-I with model membranes has been studied by intrinsic tryptophan fluorescence emission (ITFE) spectroscopy. Nine separate mutants have been created each with a unique tryptophan residue located at a different position in the interfacial recognition site (IRS) of the protein. The rapid and efficient Ca2+-independent membrane damage against unilamellar liposomes composed of DPPC/DMPA in a 9:1 molar ratio was unaffected by these substitutions. Binding studies revealed low protein affinity for these liposomes and no changes were observed in the ITFE properties. In contrast, the binding of all mutants to DPPC/DMPA liposomes in a 1:1 molar ratio was stronger, and was correlated with altered ITFE properties. The blue-shifted emission spectra and increased emission intensity of mutants at positions 31, 67 and 115-117 in the interface recognition surface of the protein suggest these regions are partially inserted into the membrane. These results are consistent with a model for the Ca2+-independent membrane damaging mechanism that involves a transient interaction of the protein with the outer phospholipid leaflet of the target membrane. (C) 2007 Elsevier Masson SAS. All rights reserved.

Portal web per a l'IES Pla d'en Boet de Mataró

Aquesta memòria tracta sobre el procediment de creació software que s’ha dut a terme per a implementar el portal web de l’IES Pla d’En Boet de Mataró, un institut públic subvencionat per la Generalitat de Catalunya. Aquest portal s’ha desenvolupat des de zero i s’ha hagut d’enllaçar amb altres aplicacions que han sigut requerides. El document conté l’anàlisi, disseny i l’implementació del portal web, i de tota la funcionalitat que l’envolta, que s’ha realitzat per satisfer els requeriments inicials. Conté, a més, les diferents anàlisis que s’han necessitat per tal d’integrar-lo amb una aplicació pròpia del centre i amb un sistema d’intercanvi de missatges, i com s’ha decidit fer-ho. S’intenta explicar alguna de les problemàtiques més importants que han aparegut al llarg del procés i que han afectat al seu desenvolupament. Les decisions preses per a resoldre-les també apareixen per avalar l’estudi realitzat. Finalment hi ha una valoració personal i una altra dels objectius aconseguits per veure que s’ha arribat a la solució final amb èxit.

Pla de gestió i conservació dels Estanyets de Pals i les Closes Fondes (Pals, Baix Empordà)

Es destaca la presència de tres espècies bioinvasores, tals com la gambúsia(Gambusiaaffinis), la canya (Arundodonax) i el cranc vermell americà (Procambarusclarkii). S’ha detectat que la qualitat de l’aigua ha disminuït en alguns paràmetres, especialment en l’Estanyet del Safareig. En els tres estanyets els sòlids dissolts totalsa TSD) estan al voltant del límit màxim recomanat per la EPA (Agència de Protecció Ambiental d’Estats Units). S’ha observat que la Cladophora, indicadora de concentracions elevades de nitrogen a l’aigua, és un cloròfit molt abundant. S’han identificat dos hàbitats d’interès comunitari no prioritari, segons la Directiva Hàbitats: les closes i les freixenedes termòfiles de Fraxinusangustifolia. En funció dels resultats obtinguts s’han elaborat les propostes de gestió i conservació per aquest espai.

Ullals de Baltasar -Delta de l'Ebre: proposta de pla de gestió i ús social

Les zones humides han sofert durant anys les velles concepcions de gestió de l'aigua, promovent la seva dessecació fins els anys 60. Com a resposta han sorgit un seguit de directives europees i lleis estatals i autonòmiques per intentar recuperar i restaurar aquests hàbitats amenaçats.

Agroecologia periurbana a Sant Cugat: document base per a la redacció del Pla de Gestió i Desenvolupament del parc rural de la Torre Negra

El parc rural de la Torre Negra ha estat protegit recentment després de 15 anys de lluita ciutadana, gràcies a l’aprovació del Pla Especial de Protecció i Millora el 29 de juny del present any. A partir d’ara, s’obre un ampli ventall de possibilitats per a la seva gestió i desenvolupament. En aquest context és on es situa el present estudi, amb la finalitat de presentar unes línies estratègiques bàsiques per a iniciar l’activitat al parc. Una activitat que té en el punt de mira el desenvolupament rural de l’espai i la transformació social de la ciutadania.

La interpretació portuguesa de Josep Pla

L’any 1975 Josep Pla publicava el número 28 de l’Obra completa: Direcció Lisboa, en què el narrador planià mostra el seu pas per tres itineraris en direcció a Portugal. El viatge per terres lusitanes esdevé una reflexió sobre la política d’Oliveira Salazar, la indústria surera i els vincles politicoculturals de Catalunya i Portugal. La interpretació portuguesa de Josep Pla aplega enfocaments i materials històrics i literaris, i vincula la poètica planiana amb la contemporaneïtat. Aquesta interpretació no es planteja com a superadora d’altres sinó com un complement, com una nova visió de Josep Pla, el narrador i l’escriptor.

Diagnosi i pla de gestió ambiental del casal de joves de Haër (Senegal)

Per tal de fer front al deteriorament i la destrucció de l’única infraestructura comuna per a joves existent al barri de Haër (M’lomp), l’associació de joves “Les Criquets de Haër” ha dut a terme un projecte de construcció d’un Casal de Joves, el qual pretén fer front no només a la manca d’infraestructures sinó també a la falta de tallers en condicions, de llocs de treball i d’espai agrícola. Amb l’objectiu de construir un centre integrat en el medi i amb capacitat per atendre les necessitats tant dels joves de Haër com d’altres poblacions properes, s’ha optat per realitzar una diagnosi ambiental del terreny on es construirà el complex. La informació obtinguda en aquesta diagnosi ha permès determinar els possibles impactes que la construcció del casal pot suposar per al medi i, a partir d’aquí, elaborar un pla de gestió ambiental dissenyant les mesures correctores més adequades per a la mitigació dels impactes detectats. Les actuacions plantejades en el Pla de gestió ambiental s’han dissenyat tenint en compte el context social i econòmic en el qual es desenvolupen les obres, així com també la predisposició de la població per a dur-les a terme i garantir-ne la continuïtat.