Local anesthetics: Interaction with human erythrocyte membranes as studied by H-1 and P-31 nuclear magnetic resonance

LOCAL ANESTHETICS: INTERACTION WITH HUMAN ERYTHROCYTE MEMBRANES AS STUDIED BY H-1 AND P-31 NUCLEAR MAGNETIC RESONANCE. The literature carries many theories about the mechanism of action of local anesthetics (LA). We can highlight those focusing the direct effect of LA on the sodium channel protein and the ones that consider the interaction of anesthetic molecules with the lipid membrane phase. The interaction between local anesthetics and human erythrocyte membranes has been studied by H-1 and P-31 nuclear magnetic resonance spectroscopy. It was found that lidocaine (LDC) and benzocaine (BZC) bind to the membranes, increase the mobility of the protons of the phospholipid's acyl chains, and decrease the mobility and/or change the structure of the polar head groups. The results indicate that lidocaine molecules are inserted across the polar and liquid interface of the membrane, establishing both electrostatic (charged form) and hydrophobic (neutral form) interactions. Benzocaine locates itself a little deeper in the bilayer, between the interfacial glycerol region and the hydrophobic core. These changes in mobility or conformation of membrane lipids could affect the Na+-channel protein insertion in the bilayer, stabilizing it in the inactivated state, thus causing anesthesia.271667

Preferential location of lidocaine and etidocaine in lecithin bilayers as determined by EPR, fluorescence and H-2 NMR

We have examined the effect of the uncharged species of lidocaine (LDC) and etidocaine (EDC) on the acyl chain moiety of egg phosphatidylcholine liposomes. Changes in membrane organization caused by both anesthetics were detected through the use of EPR spin labels (5, 7 and 12 doxyl stearic acid methyl ester) or fluorescence probes (4, 6, 10, 16 pyrene-fatty acids). The disturbance caused by the LA was greater when the probes were inserted in more external positions of the acyl chain and decreased towards the hydrophobic core of the membrane. The results indicate a preferential insertion of LDC at the polar interface of the bilayer and in the first half of the acyl chain, for EDC. Additionally, 2 H NMR spectra of multilamellar liposomes composed by acyl chain-perdeutero DMPC and EPC (1:4 mol%) allowed the determination of the segmental order (S-mol) and dynamics (T-1) of the acyl chain region. In accordance to the fluorescence and EPR results, changes in molecular orientation and dynamics are more prominent if the LA preferential location is more superficial, as for LDC while EDC seems to organize the acyl chain region between carbons 2-8, which is indicative of its positioning. We propose that the preferential location of LDC and EDC inside the bilayers creates a 'transient site', which is related to the anesthetic potency since it could modulate the access of these molecules to their binding site(s) in the voltage-gated sodium channel. (C) 2007 Elsevier B.V. All rights reserved.1321475

Interaction of benzocaine with model membranes

We measured the absorption properties, water solubility and partition coefficients (P) between n-octanol, egg phosphatidylcholine (EPC) liposomes and erythrocyte ghosts/water for benzocaine (BZC), an ester-type always uncharged local anesthetic. The interaction of BZC with EPC liposomes was followed using Electron Paramagnetic Resonance, with spin labels at different positions in the acyl chain (5, 7, 12, 16-doxylstearic acid methyl ester). Changes in lipid organization upon BZC addition allowed the determination of P values, without phase separation. The effect of BZC in decreasing membrane organization (maximum of 11.6% at approx. 0.8:1 BZC:EPC) was compared to those caused by the local anesthetics tetracaine and lidocaine. Hemolytic tests revealed a biphasic (protective/inductive) concentration-dependent hemolytic effect for BZC upon rat erythrocytes, with an effective BZC:lipid molar ratio in the membrane for protection (Re-PROT), onset of hemolysis (Re-SAT) and 100% membrane solubilization (Re-SOL) of 1.0:1, 1.1:1 and 1.3:1, respectively. The results presented here reinforce the importance of considering hydrophobic interactions in the interpretation of the effects of anesthetics on membranes. (C) 2000 Elsevier Science B.V. All rights reserved.874170021322

Ecotoxicology of Nano-TiO2 An Evaluation of its Toxicity to Organisms of Aquatic Ecosystems

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The production and use of synthetic nanoparticles is growing rapidly, and therefore the presence of these materials in the environment seems inevitable. Titanium dioxide (TiO2) presents various possible uses in industry, cosmetics, and even in the treatment of contaminated environments. Studies about the potential ecotoxicological risks of TiO2 nanoparticles (nano-TiO2) have been published but their results are still inconclusive. It should be noted that the properties of the diverse nano-TiO2 must be considered in order to establish experimental models to study their toxicity to environmentally relevant species. Moreover, the lack of descriptions and characterization of nanoparticles, as well as differences in the experimental conditions employed, have been a compromising factor in the comparison of results obtained in various studies. Therefore, the purpose of this paper is to make a simple review of the principal properties of TiO2, especially in nanoparticulate form, which should be considered in aquatic toxicology studies, and a compilation of the works that have been published on the subject.613350Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Rede NanobiotecFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação para o Desenvolvimento da UNESP (FUNDUNESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Use of Biopolymeric Membranes for Adsorption of Paraquat Herbicide from Water

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The use of membranes prepared with alginate and chitosan to adsorb paraquat aqueous solution was evaluated as a potential alternative technique for remediation of contaminated water. Production of bilayer membranes was based on the electrostatic interaction between alginate (a polyanion) and chitosan (a polycation). Herbicide adsorption experiments were performed using three different membranes, consisting of pure alginate, pure chitosan, and a chitosan/alginate bilayer. Adsorption was characterized using the Langmuir and Freundlich isotherm models, as well as by applying pseudo-first order and pseudo-second order kinetic models. The potential use of the membranes in environmental applications was evaluated using water collected from the Sorocabinha River in So Paulo State, Brazil. The results indicated that interactions between the membranes and the herbicide were strongly related to the type of biopolymer and the physical-chemical characteristics of the herbicide.223630933104Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação para o Desenvolvimento da UNESP (FUNDUNESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Spécial Mexique

Quel est le rôle de l'espace dans la structuration d'une culture ? Comment se perçoit l'influence d'une civilisation dans l'organisation par les hommes de leur espace et de leur milieu ? Un domaine fécond et encore peu exploré s'ouvre à l'analyse, lorsqu'on observe les relations entre phénomènes géographiques (espace, milieu, territoire, paysage, régions...) (Résumé d'auteur

Differential effects of uncharged aminoamide local anesthetics on phospholipid bilayers, as monitored by H-1-NMR measurements

We have collected evidences of a "transient site" for the local anesthetics (LA) lidocaine, etidocaine, bupivacaine and mepivacaine in sonicated egg phosphatidylcholine (EPC) vesicles. The effects of the uncharged anesthetic species at a fixed LA/EPC ratio inside the bilayer were measured by chemical shifts (C.S.) and longitudinal relaxation times (T-1) of the lipid hydrogens. Two sort of changes were detected: (1) decrease, indicating specific orientation of the LA aromatic ring (measured as up-field C.S. changes by the shortrange ring-current effect) and less rotational freedom (smaller T, values) for EPC hydrogens such as the two glycerol-CH2 and the choline-CH2 bound to the PO4- group, probably due to the nearby presence of the LA; (11) increase, indicating the aromatic ring is now perpendicular to the orientation observed before (causing down-field changes in C.S.) and larger T, values for all the choline and glycerol hydrogens, as a result of LA insertion behind these well-organized bilayer regions. The less hydrophobic, linear and nonlinear (lidocaine and mepivacaine, respectively) aminoamide analogs provide similar effects-described in I; their hydrophobic counterparts (etidocaine and bupivacaine) also produced comparable effects (depicted in 11). The preferential positioning and orientation of each LA inside the bilayer is then determined by its hydrophobic and steric properties. We propose that this "transient site" in the lipid milieu exists also in biological membranes, where it can modulates the access of the uncharged LA species to its site(s) of action in the voltage-gated sodium channel. © 2004 Elsevier B.V. All rights reserved.1151111

Evaluation of Cyto- and Genotoxicity of Poly(lactide-co-glycolide) Nanoparticles

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)This work reports on an analysis of the cyto- and genotoxicity of poly(lactide-co-glycolide) polymer nanoparticles, in an attempt to evaluate their mutagenic effects. Fibroblast (3T3) and human lymphocyte cell cultures were exposed to solutions containing three different concentrations of nanoparticles (5.4, 54 and 540 mu g/mL, polymer mass/volume of solution). The nanoparticles were characterized in terms of their hydrodynamic diameters, zeta potentials and polydispersity indices. The morphology of the particles was determined by atomic force microscopy. The PLGA nanospheres presented a size of 95 nm, a zeta potential of -20 mV and a spherical morphology. Cellular viability assays using fibroblast cells showed no significant alterations compared with the negative control. A cytogenetic analysis of human lymphocyte cells showed no significant changes in the mitotic index in relation to the control, indicating that in the concentration range tested, the particles used in the experimental models did not present cyto- or genotoxicity. For the tests conducted in this work we can conclude that biodegradable and biocompatible PLGA nanospheres are not toxic in the cell cultures tested (fibroblast and lymphocyte cells) and in the range of concentrations employed. The results provide new information concerning the toxic effects of particles produced using PLGA.191196202Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação para o Desenvolvimento da UNESP (FUNDUNESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq