Silver nanoparticles synthesized by the heavy metal resistant strain Amycolatopsis tucumanensis and its application in controlling red strip disease in sugarcane.

The production of bioethanol and sugar from sugarcane is an important economic activity in several countries. Sugarcane is susceptible to different phytopathogens. Over the last years, the red stripe disease caused by the bacterium Acidovorax avenae subsp. avenae produced significant losses in sugarcane crops. Bio-nanotechnology emerged as an eco-friendly alternative to the biosynthesis of antimicrobial molecules. The aims of this study were to (a) produce extracellular silver nanoparticles using the heavy metal resistant strain Amycolatopsis tucumanensis, (b) evaluate their antibacterial in vitro effect and (c) determine the potential of silver nanoparticles to protect sugarcane against red stripe disease. Amycolatopsis tucumanensis synthesized spherical silver nanoparticles with an average size of 35 nm. Nanoparticles were able to control the growth of A. avenae subsp. avenae in in vitro assays. In addition, in vivo assays in sugarcane showed a control upon the red stripe disease when silver nanoparticles were applied as preventive treatment. The Disease Severity Index was 28.94% when silver nanoparticles were applied 3 days before inoculation with A. avenae subsp. a venae. To our knowledge, this is the first report of silver nanoparticles extracellularly synthesized by an Amycolatopsis strain that were able to inhibited the growth of A. avenae subsp. avenae and control the red stripe disease in sugarcane.

Effects of mesenchymal stem cell culture on radio sterilized human amnion or radio sterilized pig skin in burn wound healing.

Deep second and third degree burns treatment requires fibroblasts, keratinocytes and other skin cells in order to grow new dermis and epidermis. Cells can proliferate, secrete growth factors and extracellular matrix required to repair the damaged tissue. Radiosterilized human amnion and radiosterilized pig skin have been used as natural origin skin dressings for burned patients. Adipose-derived mesenchymal stem cells can differentiate into fibroblasts and keratinocytes and improve wound-healing progress. These cells can stimulate vascular tissue formation, release growth factors, synthetize new extracellular matrix and immunoregulate other cells. In this study, we developed mesenchymal stem cells-cellularized skin substitutes based from radiosterilized human amnion or pig skin. Third-degree burns were induced in mice animal models to evaluate the effect of cellularized skin substitutes on burn wound healing. Mesenchymal phenotype was immunophenotypically confirmed by flow cytometry and cell viability was close to 100%. Skin recovery was evaluated in burned mice after seven and fourteen days post-coverage with cellularized and non-cellularized sustitutes. Histological techniques and immunofluorescence were used to evaluate re-epithelization and type I collagen deposition. We determined that cellularized-human amnion or cellularized-pig skin in combination with mesenchymal stem cells improve extracellular matrix deposition. Both cellularized constructs increase detection of type I collagen in newly formed mouse skin and can be potentially used as skin coverage for further clinical treatment of burned patients.

Interaction of chlorogenic acid with model lipid membranes and its influence on antiradical activity.

Chlorogenic acid (CGA) is a strong phenolic antioxidant with antibacterial properties composed by a caffeoyl ester of quinic acid. Although a number of benefits has been reported and related to interactions with the red blood cell membranes, details on its membrane action and how composition and membrane state may affect it, is not yet well defined. In this work, the interaction of CGA with lipid monolayers and bilayers composed by 1,2-dimiristoyl-sn-glycero-3-phosphocholine (DMPC); 1,2-di-O-tetradecyl-sn-glycero-3-phosphocholine (14:0 diether PC); 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-di-O-hexadecyl-sn-glycero-3-phosphocholine (16:0 diether PC) were studied at different surface pressures (π). The kinetics of interaction was found to be more rapid in DMPC than in the absence of carbonyl groups. Measurements by FTIR-ATR at different water activities confirm specific interactions of CGA with carbonyl and phosphate groups affecting water level along hydrocarbon region. The antioxidant activity of CGA in the presence of DMPC unilamellar vesicles, evidenced by the absorbance reduction of the radical cation ABTS•+, is significantly different with respect to aqueous solution. The influence of CGA on antiradical activity (ARA) with lipid membranes depending on the hydration state of the lipid interface is discussed.

Effect of cholesterol on the hydration properties of ester and ether lipid membrane interphases.

Fluorescence spectroscopy and Molecular Dynamics results show that cholesterol reduces water along the chains in ether lipids by changing the water distribution pattern between tightly and loosely bound water molecules. Water distribution was followed by emission spectra and generalized polarization of 6-dodecanoyl-2-dimethyl aminonaphthalene (Laurdan) inserted in 1,2-dimiristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2-di-O-tetradecyl-sn-glycero-3-phosphocholine (14: 0 Diether PC) membranes. Molecular Dynamics simulations indicate that the action of cholesterol could be different in ether PC in comparison to ester PC. In addition, Cholesterol seems to act "per se" as an additional hydration center in ether lipids. Regardless of the phase state, cholesterol both in DMPC and 14:0 Diether PC vesicles, changed the distribution of water molecules decreasing the dipole relaxation of the lipid interphase generating an increase in the non-relaxable population. Above 10% Cholesterol/14:0 Diether PC ratio vesicles' interphase present an environment around Laurdan molecules similar to that corresponding to ester PC.

Breakdown of classical paradigms in relation to membrane structure and functions.

Updates of the mosaic fluid membrane model implicitly sustain the paradigms that bilayers are closed systems conserving a state of fluidity and behaving as a dielectric slab. All of them are a consequence of disregarding water as part of the membrane structure and its essential role in the thermodynamics and kinetics of membrane response to bioeffectors. A correlation of the thermodynamic properties with the structural features of water makes possible to introduce the lipid membrane as a responsive structure due to the relaxation of water rearrangements in the kinetics of bioeffectors' interactions. This analysis concludes that the lipid membranes are open systems and, according to thermodynamic of irreversible formalism, bilayers and monolayers can be reasonable compared under controlled conditions. The inclusion of water in the complex structure makes feasible to reconsider the concept of dielectric slab and fluidity.

Water Behavior at the Phase Transition of Phospholipid Matrixes Assessed by FTIR Spectroscopy.

Lipid membranes are one of the most important biological matrixes in which biochemical processes take place. This particular lipid arrangement is driven by different water disposition interacting with it, which is related to different water states with different energy levels at the interphase. In our work, we report changes in water content and distinctive water states by Fourier transform infrared (FTIR) spectroscopy of this self-assembled matrix at different water contents and temperatures. To determine whether water properties at lipid interphases depend on the group of the lipid molecule at which it is bound the phase-transition temperature of 1,2-dimiristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-di-O-tetradecyl-sn-glycero-3-phosphocholine (14:0 diether PC) was followed by the changes in frequency of the different groups of the lipids by attenuated total reflection (ATR)-FTIR spectroscopy at different humidities. A comparison of these two lipids enables us to put into relevance the contribution of the CO groups as a hydration site. These changes were compared with those occurring at the water band, and a value of the enthalpic change was evaluated from them. The -OH stretching in the liquid water IR spectrum is the principal region used to understand its molecular organization (4000-3000 cm-1). The strength of hydrogen bonding depends on the cooperative/anticooperative nature of the surrounding hydrogen bonds, with the strongest hydrogen bonds giving the lowest vibrational frequencies. Thus, we can use water as a mirror of the membrane state in this kind of biological systems. Different phospholipids associate water at particular modes according to their structures. This may produce modulation of packing and hydration suitable for the incorporation of amino acids, peptides, and enzymes.

Modulation of Interfacial Hydration by Carbonyl Groups in Lipid Membranes.

The lack of carbonyl groups and the presence of ether bonds give the lipid interphase a different water organization around the phosphate groups that affects the compressibility and electrical properties of lipid membranes. Generalized polarization of 1,2-di-O-tetradecyl-sn-glycero-3-phosphocholine (14:0 diether PC) in correlation with Fourier transform infrared (FTIR) analysis indicates a higher level of polarizability of water molecules in the membrane phase around the phosphate groups both below and above Tm. This reorganization of water promotes a different response in compressibility and dipole moment of the interphase, which is related to different H bonding of water molecules with phosphates (PO) and carbonyl (CO) groups.

Effect of cholesterol on the surface polarity and hydration of lipid interphases as measured by Laurdan fluorescence: New insights.

Comparison of the behavior of Laurdan in gel and in the liquid crystalline DPPC bilayers with that observed in chloroform and OctOH allow concluding that changes in the membrane lipid order cannot be ascribed to changes in viscosity of the local environment. Cholesterol acts as a spacer below the transition temperature of DPPC, promoting a disorder state in the acyl chain region. No evidence of water entrance has been detected with Laurdan up to 30% Cholesterol in DPPC in this condition. In contrast, Chol displaces to longer values the wavelength of Laurdan in membranes in the liquid crystalline state. This decrease in polarity occurs above 5% Chol and is directly related to the water extrusion produced by Chol. This effect is similar to that occurring in liquid crystalline membranes subjected to hypertonic stress. The behavior is comparable to that of Laurdan in OctOH at different water ratios below 5% Chol/DPPC. At higher ratios, other changes are evident.

Silver-pig skin nanocomposites and mesenchymal stem cells: suitable antibiofilm cellular dressings for wound healing.

BACKGROUND: Treatment of severe or chronic skin wounds is an important challenge facing medicine and a significant health care burden. Proper wound healing is often affected by bacterial infection; where biofilm formation is one of the main risks and particularly problematic because it confers protection to microorganisms against antibiotics. One avenue to prevent bacterial colonization of wounds is the use of silver nanoparticles (AgNPs); which have proved to be effective against non-multidrug-resistant and multidrug-resistant bacteria. In addition, the use of mesenchymal stem cells (MSC) is an excellent option to improve wound healing due to their capability for differentiation and release of relevant growth factors. Finally, radiosterilized pig skin (RPS) is a biomatrix successfully used as wound dressing to avoid massive water loss, which represents an excellent carrier to deliver MSC into wound beds. Together, AgNPs, RPS and MSC represent a potential dressing to control massive water loss, prevent bacterial infection and enhance skin regeneration; three essential processes for appropriate wound healing with minimum scaring. RESULTS: We synthesized stable 10 nm-diameter spherical AgNPs that showed 21- and 16-fold increase in bacteria growth inhibition (in comparison to antibiotics) against clinical strains Staphylococcus aureus and Stenotrophomonas maltophilia, respectively. RPS samples were impregnated with different AgNPs suspensions to develop RPS-AgNPs nanocomposites with different AgNPs concentrations. Nanocomposites showed inhibition zones, in Kirby-Bauer assay, against both clinical bacteria tested. Nanocomposites also displayed antibiofilm properties against S. aureus and S. maltophilia from RPS samples impregnated with 250 and 1000 ppm AgNPs suspensions, respectively. MSC were isolated from adipose tissue and seeded on nanocomposites; cells survived on nanocomposites impregnated with up to 250 ppm AgNPs suspensions, showing 35% reduction in cell viability, in comparison to cells on RPS. Cells on nanocomposites proliferated with culture days, although the number of MSC on nanocomposites at 24 h of culture was lower than that on RPS. CONCLUSIONS: AgNPs with better bactericide activity than antibiotics were synthesized. RPS-AgNPs nanocomposites impregnated with 125 and 250 ppm AgNPs suspensions decreased bacterial growth, decreased biofilm formation and were permissive for survival and proliferation of MSC; constituting promising multi-functional dressings for successful treatment of skin wounds.

Water populations in restricted environments of lipid membrane interphases.

We employ molecular dynamics simulations to study the hydration properties of Dipalmitoylphosphatidylcholine (DPPC) bilayers, both in the gel and the liquid crystalline states. We show that while the tight hydration centers (PO and CO moieties) are significantly hydrated in both phases, the gel-fluid transition involves significant changes at the second hydration shell, particularly at the buried region between the hydrocarbon tails. Thus, while almost no buried water population exists in the gel state below the carbonyls, this hydrophobic region becomes partially water accesible in the liquid crystalline state. We shall also show that such water molecules present a lower H-bond coordination as compared to the molecules at the primary hydration shell. This means that, while the latter are arranged in relatively compact nanoclusters (as already proposed), the buried water molecules tend to organize themselves in less compact structures, typically strings or branched strings, with a scarce population of isolated molecules. This behavior is similar to that observed in other hydration contexts, like water penetrating carbon nanotubes or model hydrophobic channels or pores, and reflects the reluctance of water to sacrifice HB coordination.

Interaction of Phenylalanine with DPPC Model Membranes: More Than a Hydrophobic Interaction.

The negative free energy previously reported is explained by the stabilization of a PC-Phe (phosphocholine-phenylalanine) complex in the presence of water shown by the decrease in the symmetric stretching frequency of the phosphate group of the lipid (PO2(-)). An entropic contribution due to the disruption of the water network around the phenyl and in the membrane defect may be invoked. The dipole potential decrease is explained by the orientation of the carboxylate opposing to the CO of the lipids with oxygen moiety toward the low hydrated hydrocarbon core. The symmetric bending frequency of NH3(+) group of Phe, decreases in 5.2 cm(-1) in relation to water congruent with zeta potential shift to positive values. The Phe to DPPC dissociation constant is Kd = 2.23 ± 0.09 mM, from which the free energy change is about -4.54 kcal/mol at 25 °C. This may be due to hydrophobic contributions and two hydrogen bonds.

Phenylalanine interaction with lipid monolayers at different pHs.

The influence of Phe on the surface pressure of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) monolayers at the air-water interface was studied at different initial surface pressures (26 and 40 mN/m) and two pHs (5.0 and 7.3) at constant temperature (20 °C). Changes produced by the aminoacid added to the subphase on the surface pressure and on the dipole potential of lipid monolayers were measured at a fixed area. Compressibility properties of the monolayers at different pHs were studied by (π-A) isotherms. The results suggest that Phe intercalates into a DPPC film at the air-water interface at pH 5 and forms a different arrangement at pH 7.3. The possible relevance of these results of the effect of Phe in physiological conditions is discussed.

Surface and hysteresis properties of lipid interphases composed by head group substituted phosphatidylethanolamines.

This work analyzes the surface properties of PE-containing membranes modified at the head group region by the addition of methyl and ethyl residues at or near the amine group. These residues alter the lipid-lipid and lipid-water interactions by changes in the hydrogen bonding capability and the charge density of the amine group thus affecting the electrostatic interaction. The results obtained by measuring the dipole potential, the zeta potential, the area per lipid and the compressibility properties allow to conclude that the H-bonding capability prevails in the lipid-lipid interaction. The non polar groups attached to the C2-carbon of the ethanolamine chain introduces a steric hindrance against compression and increases the dipole potential. The analysis of areas suggests that lipids with methylated head groups have a much larger compressibility at expense of the elimination of hydration water, which is congruent with the broader extent of the hysteresis loop.

Connected and isolated CH2 populations in acyl chains and its relation to pockets of confined water in lipid membranes as observed by FTIR spectrometry.

Analysis of the band corresponding to the frequency of vibrational symmetric stretching mode of methylene groups in the lipid acyl chains and the bands of water below and above the phase transition of different lipids by Fourier transform infrared spectroscopy gives strong support to the formation of confined water pockets in between the lipid acyl chains. Our measures and analysis consolidate the mechanism early proposed by Traüble, in the sense that water is present in kinks formed by trans-gauche isomers along the hydrocarbon tails. The formation of these regions depends on the acyl lipid composition, which determines the presence of different populations of water species, characterized by its degree of H bond coordination in fluid saturated or unsaturated lipids. The free energy excess due to the reinforcement of the water structure along few water molecules in the adjacencies of exposed membrane residues near the phase transition is a reasonable base to explain the insertion and translocation of polar peptides and amino acid residues through the biomembrane on thermodynamic and structural grounds.

Water state and carbonyl distribution populations in confined regions of lipid bilayers observed by FTIR spectroscopy.

It has been suggested that water in confined regions presents different properties than bulk water, mainly because of the changes in water population species that may be induced by the adjacent walls of different polarities in terms of hydrogen bond formation. In this context, it would be expected that lipids in the gel and the fluid states should offer different templates for water organization. The presence of water pockets or defects in lipid bilayers has been proposed to explain the insertion of charged and polar peptides and amino acids in membranes. In this work, we provide direct evidence by means of FTIR spectroscopy that water band profiles are changed whether lipids are in the solid state, in the gel state after heating and cooling across the phase transition, or in the fluid state. The different bands found in each case were assigned to different H-bonded water populations in agreement with the exposure of carbonyl groups.

Effect on seroprevalence of anti-poliovirus antibodies and on vaccination coverage of the implementation of a DTwP-IPV-Hib vaccination programme in a South American city.

Vaccination coverage and seroprevalence of poliovirus antibodies were assessed in Argentinean children (aged 8-12 and 19-21 months) living in Cordoba City pre-/post-implementation of a DTwP-IPV-Hib vaccination programme, and compared to those of controls from neighbouring populations receiving a full oral poliovirus vaccine schedule. Vaccination coverage was higher in control areas pre-intervention; this increased post-intervention in Cordoba (>90%) but not in control areas. Poliovirus types 1 and 2 seroprotection rates were >97% in all groups pre-/post-intervention. Type 3 seroprotection rates were generally lower, but increased post-intervention in Cordoba becoming significantly higher than control rates. Anti-type 1 and 3 antibody titres increased twofold and sevenfold, respectively, post-intervention, whereas anti-type 2 antibody titres decreased ~40% in the 8-12 months group. All titres increased in the 19-21 months post-intervention group. The introduction of a three-dose primary DTwP-IPV-Hib schedule maintained protection against poliovirus types 1 and 2, and increased protection against type 3, while vaccine coverage in the study area increased.

Nematoides entomopatogênicos infectam ovos e adultos cigarrinha-das-pastagens? / Can entomopathogenic nematodes infect eggs and adults of spittlebugs?

Nematoides entomopatogênicos (NEPs) são potenciais controladores biológicos das ninfas de cigarrinhas-das-pastagens. Com o objetivo de avaliar a eficiência dos NEPs contra as demais fases de vida (ovos e adultos) de Mahanarva spectabilis, desenvolveu-se experimento sob condições de laboratório. Ovos em diferentes fases de desenvolvimento embrionário foram expostos aos NEPs Steinernema anomali. S. carpocapsae. S. feltiae. S. riobrave. Heterorhabditis amazonensis RSC1, H. amazonensis RSC5, Heterorhabditis sp. JPM3, Heterorhabditis indica LPP1 e Heterorhabditis sp. PI no volume de 2 mL com concentração de 2.000 JIs/mL. A eficiência do patógeno foi avaliada através da dissecação dos ovos após sete dias de exposição. No experimento com adultos as unidades experimentais (UEs) consistiram de gaiolas contendo adultos da cigarrinha submetidas a uma suspensão aquosa com 20 mL contendo 8.000 JIs/mL de S. riobrave (tratamento) das quais se compararam a mortalidade e número de ovos recuperados com os mesmos parâmetros das UEs livres de nematoides (testemunha). Os nematoides não foram capazes de penetrar os ovos do cercopídeo. Não foi observada morte de indivíduos adultos de cigarrinha, tampouco se observou redução no número de ovos entre as UEs tratadas e testemunhas. Não houve influência da presença de nematoides sobre a freqüência de adultos que permaneciam sobre o substrato. Não foi caracterizada vulnerabilidade destas fases de desenvolvimento do inseto às espécies testadas desses patógenos, portanto, não devem ser aplicados visando o controle dessas fases de desenvolvimento do inseto.

Entomopathogenic nematodes (EPNs) are potential biological controllers of spittlebug nymphs. With the aim to evaluate the efficiency of EPNs against the other life stages (eggs and adults) of Mahanarva spectabilis, an assay was developed in laboratory conditions. Eggs at different embrionary development phases were exposed to the EPNs Steinernema anomali. S. carpocapsae. S. feltiae. S. riobrave. Heterorhabditis amazonensis RSC1, H. amazonensis RSC5, Heterorhabditis sp. JPM3, Heterorhabditis indica LPP1 and Heterorhabditis sp. PI in a 2 mL aqueous suspension of 2,000 IJ/mL concentration. The efficiency of the pathogens was evaluated by way of egg desiccation after 7 days. In the assay with adults the experimental units consisted of cages containing spittlebug adults exposed to an aqueous suspension (20 mL) with 8,000 IJs/mL of Steinernema riobrave (treatment), and mortality and recovered eggs number were compared with the same parameters of cages free of nematodes (control group). No individual mortality was observed, nor was there a reduction in recovered egg number between the treated and control cages. The presence of nematodes presented no influence on the adults staying on the substrate. Thus, there was no vulnerability in these stages of development of the insect to these pathogens, and therefore it should not be applied for the control of these insect development phases.

NEMATOIDES ENTOMOPATOGÊNICOS INFECTAM OVOS E ADULTOS CIGARRINHA-DAS-PASTAGENS?

ABSTRACT Entomopathogenic nematodes (EPNs) are potential biological controllers of spittlebug nymphs. With the aim to evaluate the efficiency of EPNs against the other life stages (eggs and adults) of Mahanarva spectabilis, an assay was developed in laboratory conditions. Eggs at different embrionary development phases were exposed to the EPNs Steinernema anomali. S. carpocapsae. S. feltiae. S. riobrave. Heterorhabditis amazonensis RSC1, H. amazonensis RSC5, Heterorhabditis sp. JPM3, Heterorhabditis indica LPP1 and Heterorhabditis sp. PI in a 2 mL aqueous suspension of 2,000 IJ/mL concentration. The efficiency of the pathogens was evaluated by way of egg desiccation after 7 days. In the assay with adults the experimental units consisted of cages containing spittlebug adults exposed to an aqueous suspension (20 mL) with 8,000 IJs/mL of Steinernema riobrave (treatment), and mortality and recovered eggs number were compared with the same parameters of cages free of nematodes (control group). No individual mortality was observed, nor was there a reduction in recovered egg number between the treated and control cages. The presence of nematodes presented no influence on the adults staying on the substrate. Thus, there was no vulnerability in these stages of development of the insect to these pathogens, and therefore it should not be applied for the control of these insect development phases.

RESUMO Nematoides entomopatogênicos (NEPs) são potenciais controladores biológicos das ninfas de cigarrinhas-das-pastagens. Com o objetivo de avaliar a eficiência dos NEPs contra as demais fases de vida (ovos e adultos) de Mahanarva spectabilis, desenvolveu-se experimento sob condições de laboratório. Ovos em diferentes fases de desenvolvimento embrionário foram expostos aos NEPs Steinernema anomali. S. carpocapsae. S. feltiae. S. riobrave. Heterorhabditis amazonensis RSC1, H. amazonensis RSC5, Heterorhabditis sp. JPM3, Heterorhabditis indica LPP1 e Heterorhabditis sp. PI no volume de 2 mL com concentração de 2.000 JIs/mL. A eficiência do patógeno foi avaliada através da dissecação dos ovos após sete dias de exposição. No experimento com adultos as unidades experimentais (UEs) consistiram de gaiolas contendo adultos da cigarrinha submetidas a uma suspensão aquosa com 20 mL contendo 8.000 JIs/mL de S. riobrave (tratamento) das quais se compararam a mortalidade e número de ovos recuperados com os mesmos parâmetros das UEs livres de nematoides (testemunha). Os nematoides não foram capazes de penetrar os ovos do cercopídeo. Não foi observada morte de indivíduos adultos de cigarrinha, tampouco se observou redução no número de ovos entre as UEs tratadas e testemunhas. Não houve influência da presença de nematoides sobre a freqüência de adultos que permaneciam sobre o substrato. Não foi caracterizada vulnerabilidade destas fases de desenvolvimento do inseto às espécies testadas desses patógenos, portanto, não devem ser aplicados visando o controle dessas fases de desenvolvimento do inseto.

Structural and dynamical surface properties of phosphatidylethanolamine containing membranes.

The hydration of solid dimyristoylphosphatidylethanolamine (DMPE) produces a negligible shift in the asymmetric stretching frequency of the phosphate groups in contrast to dimyristoylphosphatidylcholine (DMPC). This suggests that the hydration of DMPE is not a consequence of the disruption of the solid lattice of the phosphate groups as occurs in DMPC. The strong lateral interactions between NH(3) and PO(2)(-) groups present in the solid PEs remain when the lipids are fully hydrated and seem to be a limiting factor for the hydration of the phosphate group hindering the reorientation of the polar heads. The lower mobility is reflected in a higher energy to translocate the phosphoethanolamine (P-N) dipoles in an electrical field. This energy is decreased in the presence of increasing ratios of PCs of saturated chains in phosphoethanolamine monolayer. The association of PC and PE in the membrane affecting the reorientation of the P-N groups is dependent of the chain-chain interaction. The dipole potentials of PCs and PEs mixtures show different behaviors according to the saturation of the acyl chain. This was correlated with the area in monolayers and the hydration of the P-N groups. In spite of the low hydration, DMPE is still able to adsorb fully hydrated proteins, although in a lower rate than DMPC at the same surface pressure. This indicates that PE interfaces possess an excess of surface free energy to drive protein interaction. The relation of this free energy with the low water content is discussed.

Structural and functional properties of hydration and confined water in membrane interfaces.

The scope of the present review focuses on the interfacial properties of cell membranes that may establish a link between the membrane and the cytosolic components. We present evidences that the current view of the membrane as a barrier of permeability that contains an aqueous solution of macromolecules may be replaced by one in which the membrane plays a structural and functional role. Although this idea has been previously suggested, the present is the first systematic work that puts into relevance the relation water-membrane in terms of thermodynamic and structural properties of the interphases that cannot be ignored in the understanding of cell function. To pursue this aim, we introduce a new definition of interphase, in which the water is organized in different levels on the surface with different binding energies. Altogether determines the surface free energy necessary for the structural response to changes in the surrounding media. The physical chemical properties of this region are interpreted in terms of hydration water and confined water, which explain the interaction with proteins and could affect the modulation of enzyme activity. Information provided by several methodologies indicates that the organization of the hydration states is not restricted to the membrane plane albeit to a region extending into the cytoplasm, in which polar head groups play a relevant role. In addition, dynamic properties studied by cyclic voltammetry allow one to deduce the energetics of the conformational changes of the lipid head group in relation to the head-head interactions due to the presence of carbonyls and phosphates at the interphase. These groups are, apparently, surrounded by more than one layer of water molecules: a tightly bound shell, that mostly contributes to the dipole potential, and a second one that may be displaced by proteins and osmotic stress. Hydration water around carbonyl and phosphate groups may change by the presence of polyhydroxylated compounds or by changing the chemical groups esterified to the phosphates, mainly choline, ethanolamine or glycerol. Thus, surface membrane properties, such as the dipole potential and the surface pressure, are modulated by the water at the interphase region by changing the structure of the membrane components. An understanding of the properties of the structural water located at the hydration sites and the functional water confined around the polar head groups modulated by the hydrocarbon chains is helpful to interpret and analyze the consequences of water loss at the membranes of dehydrated cells. In this regard, a correlation between the effects of water activity on cell growth and the lipid composition is discussed in terms of the recovery of the cell volume and their viability. Critical analyses of the properties of water at the interface of lipid membranes merging from these results and others from the literature suggest that the interface links the membrane with the aqueous soluble proteins in a functional unit in which the cell may be considered as a complex structure stabilized by water rather than a water solution of macromolecules surrounded by a semi permeable barrier.