Interaction of cysteine and its derivatives with monolayers of dipalmitoylphosphatidylcholine.

Molecular interactions between l-cysteine (Cys) and its ester derivatives (Cysx); l-cysteine ethyl ester (CE), l-cysteine methyl ester (CM) and N-acetyl l-cysteine (NAC) with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) monolayers were investigated using Langmuir film balance technique. The effect of charge on monolayers made of cysteine and three ester derivatives with DPPC was investigated by working with un-buffered and buffered subphases. Also, the effects of cysteine derivatives interaction with DPPC monolayers were studied measuring the change in the surface tension upon aminoacid injection in the subphase whilst keeping lipid molecular density and lateral packing controlled. Cysteine and its ester derivatives showed interfacial activity reducing the air/water surface tension (πi) by 4 mN m-1. However, ester derivatives were able to insert into preformed DPPC monolayers at much higher surface pressures (Δπ), indicating a preferential interaction of Cysx with DPPC. The results indicate that, although the different derivatives of cysteine presented low surface activity, they were able to favourably interact with DPPC monolayers. Also, compression isotherms experiments in binary mixtures indicate that the more surface active compounds stabilized the gel phase of DPPC. The charge on cysteine and its derivatives did not increase the observed effects.

Interactions of valproic acid with lipid membranes of 1,2-dimyristoyl-sn-glycero-3-phosphocholine.

Lipid bilayers of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) were prepared in two forms, as a suspension of multilamellar spherical vesicles and as planar membranes deposited on a conductive solid support. We used Fourier Transformed Infrared (FTIR) and Raman spectroscopic techniques to study the lipid vesicles while the solid supported bilayers were characterized by using electrochemical experiments (cyclic voltammetry and impedance). Valproic acid (Valp) was either present in the solution or incorporated into the lipid structure. As the Valp:DMPC ratio increases the phase transition temperature decreases while the phase transition becomes less marked. Moreover, for the Valp:DMPC complex species a slight decrease in the number of gauche isomers was observed relative to the number of trans isomers what corresponds to an increase in the packing density of the acylic chains. Based on derived electrical properties of the supported membranes it can be concluded that Valp induces the formation of pores and other defects in the lipid films. Valp incorporated into the membrane is seriously detrimental to the bilayer stability.

Characterization of interactions of eggPC lipid structures with different biomolecules.

In this paper we study the interactions of two biomolecules (ascorbic acid and Annonacin) with a bilayer lipid membrane. Egg yolk phosphatidylcholine (eggPC) liposomes (in crystalline liquid state) were prepared in solutions of ascorbic acid (AA) at different concentration levels. On the other hand, liposomes were doped with Annonacin (Ann), a mono-tetrahydrofuran acetogenin (ACG), which is an effective citotoxic substance. While AA pharmacologic effect and action mechanisms are widely known, those of Ann's are only very recently being studied. Both Fourier Transformed Infrared (FTIR) and Raman spectroscopic techniques were used to study the participation of the main functional groups of the lipid bilayer involved in the membrane-solution interaction. The obtained spectra were comparatively analyzed, studying the spectral bands corresponding to both the hydrophobic and the hydrophilic regions in the lipid bilayer. Electrochemical experiments namely; impedance spectroscopy (EIS) and cyclic voltamperometry (CV) were used as the main characterization techniques to analyse stability and structural changes of a model system of supported EggPC bilayer in connection with its interactions with AA and Ann. At high molar ratios of AA, there is dehydration in both populations of the carbonyl group of the polar head of the lipid. On the other hand, Ann promotes the formation of hydrogen bonds with the carbonyl groups. No interaction between AA and phosphate groups is observed at low and intermediate molar ratios. Ann is expected to be able to induce the dehydration of the phosphate groups without the subsequent formation of H bonds with them. According to the electrochemical analysis, the interaction of AA with the supported lipid membrane does not alter its dielectric properties. This fact can be related to the conservation of structured water of the phosphate groups in the polar heads of the lipid. On the other hand, the incorporation of Ann into the lipid membrane generates an increase in the number of defects while changes the dielectric constant. This, in turn, can be associated with the induced dehydration of the phosphate groups.

Evaluation of the physical properties and conservation of the antioxidants content, employing inulin and maltodextrin in the spray drying of blueberry juice.

In this work, two carbohydrate polymers, inulin (I) and maltodextrin (MX), were compared as carrying agents in the spray drying of blueberry juice (BJ). The physicochemical properties and the conservation of the antioxidants content were characterized. Both systems, showed non-agglomerated particles and light-purple color appearance. Powders were subjected to the adsorption of water, and the glass transition temperature (Tg) decreased with the water activity. The evolution of the microstructure in the MX-BJ remained unchanged, while the I-BJ presented an abrupt change from amorphous to crystalline. This was corroborated by scanning electron microscopy (SEM), observing in the I-BJ system, the change from spherical into irregular shape particles. In the conservation of the antioxidants content, the MX-BJ showed a better performance. Anyhow, the performance of both carbohydrate polymers as carrying agents in the spray drying of BJ was effective.

Vibrational and structural behavior of (L)-cysteine ethyl ester hydrochloride in the solid state and in aqueous solution.

The aim of this work is to evaluate the vibrational and structural properties of l-cysteine ethyl ester hydrochloride (CE), and its electronic behavior mainly in relation to the action of the thiol and amine groups at different degrees of solvation. The crystal structure of CE was determined at room temperature by X-ray diffraction methods. Infrared and Raman spectra were collected to compare the behavior of different functional groups in the molecule, both in the solid phase and in aqueous solution. Its UV and circular dichroism spectra were also measured in aqueous solution. The influence of an aqueous environment on the CE spectra was simulated by means of implicit (polarizable continuum model) and explicit (molecular dynamics, solute-solvent clusters) methods. Calculations in explicit and continuous solvent are of interest to explain the behavior of bioavailable sites in this medium. The study was completed by natural bond orbital analysis to determine the presence of hyperconjugative interactions.

Lysophosphatidylcholine-arbutin complexes form bilayer-like structures.

Arbutin is known to suppress melanin production in murine B16 melanoma cells and inhibit phospholipase action. This encourages the possibility to stabilize it in lipid aggregates for its administration in medical applications. Thus, it was of interest to demonstrate that monomyristoylphosphatidylcholine (14:0 lysoPC) and arbutin may form association complexes. This was studied by Electron Microscopy (EM), 31P Nuclear Magnetic Resonance (31P NMR), Electronic Paramagnetic Resonance (EPR) and Fourier Transform Infrared Spectroscopy (FTIR). EM images show the formation of particles of c.a. 6 nm in diameter. For a 1:1 lysoPC-arbutin molar ratio 31P NMR shows a spectrum with a shoulder that resembles the axially symmetric spectrum characteristic of vesicles. The addition of La3+ ions to the arbutin-lysoPC complex allows one to distinguish two phosphorous populations. These results suggest that arbutin-lysoPC forms vesicles with bilayers stabilized in an interdigitated array. FTIR spectroscopy shows that arbutin interacts with the hydrated population of the carbonyl groups and with the phosphates through the formation of hydrogen bonds. It is interpreted that hydrophobic interactions among the phenol group of arbutin and the acyl chain of lysoPC are responsible for the decrease in acyl chain mobility observed at the 5th C level by EPR. A model proposing the formation of interdigitated bilayers of arbutin-lysoPC could explain the experimental results.

Arbutin blocks defects in the ripple phase of DMPC bilayers by changing carbonyl organization.

The effect of arbutin, a 4-hydroxyphenyl-beta-glucopyranoside, on dimyristoylphosphatidylcholine (DMPC) bilayers was studied by turbidimetry, EPR and FTIR spectroscopies. The disruption of DMPC multilamellar vesicles (MLV's) with monomyristoylphosphatidylcholine (lysoPC), a product of hydrolysis of phospholipase A(2) (PLA(2)), is more efficient at 18 degrees C, where DMPC MLV's are known to be in the ripple P(beta') phase, than at 10 degrees C (L(beta') flat gel phase). Disruption at 18 degrees C was inhibited by increasing concentrations of arbutin in the solution. This inhibition was correlated with the disappearance of the ripple phase in MLV's when arbutin is present. Shifts in FTIR carbonyl bands caused by arbutin or by temperature changes allow us to propose a model. It is interpreted that the changes in the water-hydrocarbon interface caused by arbutin, forcing a reaccommodation of the carbonyl groups, eliminate the topological defects in the lattice due to mismatches among regions with different area per lipid where lysoPC can insert.

Experimental and theoretical study of the hydration of phosphate groups in esters of biological interest.

We have studied the influence of different groups esterified to phosphates on the strength of the interaction of the PO bond with one water molecule. Experimental vibrational spectra of PO(4)3-, HPO4(2-), H2PO4-, phosphoenolpiruvate (PEP) and ortho-phosphocholamine (o-PC) were obtained by means of FTIR spectroscopy. Geometry calculations were performed using standard gradient techniques and the default convergence criteria as implemented in GAUSSIAN 98 Program. In order to assess the behaviour of such DFT theoretical calculations using B3LYP with 6-31G* and 6-311++G** basis sets, we carried out a comparative work for those compounds. The results were then used to predict the principal bands of the vibrational spectra and molecular parameters (geometrical parameters, stabilisation energies, electronic density). In this work, the relative stability and the nature of the PO bond in those compounds were systematically and quantitatively investigated by means of Natural Bond Order (NBO) analysis. The topological properties of electronic charge density are analysed employing Bader's Atoms in Molecules theory (AIM). The hydrogen bonding of phosphate groups with water is highly stable and the PO bond wavenumbers are shifted to lower experimental and calculated values (with the DFT/6-311++G** basis set). Accordingly, the predicted order of the relative stability of the hydrogen bonding of the water molecule to the PO bond of the investigated compounds is: PO(4)3->HPO4(2-)>H2PO4->phosphoenolpiruvate>phosphocholamine for the two basis sets used.

Hydration of inorganic phosphates in crystal lattices and in aqueous solution. An experimental and theoretical study.

B3LYP/6-31G* and 6-311++G** calculations have been carried out in order to study the hydration of phosphates in aqueous media. Optimized geometries and relative stabilities for PO4(-3), HPO(4)-2, H2PO4(-1) have been calculated considering the interaction with one, two, three, four and five discrete water molecules and taking into account the solvent effect by using the self-consistent reaction field theory (Onsager and PCM methods). The role of specific and bulk contributions of solvent effect on the observable properties of phosphate compounds is analysed. Good agreement between theoretical and available experimental results of harmonic vibration frequencies is found. Significant effects on the geometrical and vibrational frequencies are found for those studied phosphate anions. The results presented here provide a first step toward the understanding of the phosphate group as a hydration sensor in lipid bilayers.

FTIR analysis of the interaction of arbutin with dimyristoyl phosphatidylcholine in anhydrous and hydrated states.

In this paper, the interaction of arbutin with dimyristoylphosphatidylcholine bilayers was studied by FTIR spectrometry. The results show that arbutin interacts in different extents with the phosphate and carbonyl groups of membranes in the gel state, the liquid crystalline state or subjected to osmotic stress. The effect, in the presence of water, on the antisymmetric stretching of the phosphate groups is qualitatively similar to that found with other molecules composed by a glucose moiety such as trehalose and sucrose. However, significant differences were found between these compounds and arbutin in the carbonyl region. Arbutin displaces the PO2- antisymmetric stretching to lower frequencies in lipids dispersed in water. This indicates strong hydrogen bonding. In contrast, in the solid state, this frequency increases. The effect on the carbonyl groups varies depending on the hydration state of the bilayer, which is achieved by changing the phase state of the bilayer or by osmotic stress. The hydrocarbon region is not affected by arbutin in the excess of water. However, symmetric and antisymmetric stretching of CH2 and CH3 are strongly affected in the dry state.

Dehydration of carbonyls and phosphates of phosphatidylcholines determines the lytic action of lysoderivatives.

The purpose of this study was to correlate the effectiveness of the lysoPC to disrupt bilayers with the effects of trehalose and sucrose on the hydration sites of a lipid bilayer. The vibration frequencies of carbonyls and phosphates was measured at 18 degrees C for different ratios of monomyristoylphosphatidylcholine and dimyristoylphosphatidylcholine vesicles prepared in water, sucrose and trehalose. The disruption point of the bilayer, evaluated by following the changes in the turbidity of the suspension of unilamellar vesicles, was decreased when the vesicles were prepared in 100 mM sucrose. The increase of the lytic action is directly related to the extent of hydration of the carbonyl populations. It is interpreted that the insertion of the sucrose molecule in the interface causes local changes in interfacial structure, such as the dehydration of the second population of the carbonyls that may be identified as defects of packing. In contrast, the insertion of trehalose by replacing water simultaneously at the carbonyls and the phosphates does not cause defects of packing. For this reason, the lytic action is produced at a concentration very similar to that found in water.

Ozone depletion and UVB radiation: impact on plant DNA damage in southern South America.

The primary motivation behind the considerable effort in studying stratospheric ozone depletion is the potential for biological consequences of increased solar UVB (280-315 nm) radiation. Yet, direct links between ozone depletion and biological impacts have been established only for organisms of Antarctic waters under the influence of the ozone "hole;" no direct evidence exists that ozone-related variations in UVB affect ecosystems of temperate latitudes. Indeed, calculations based on laboratory studies with plants suggest that the biological impact of ozone depletion (measured by the formation of cyclobutane pyrimidine dimers in DNA) is likely to be less marked than previously thought, because UVA quanta (315-400 nm) may also cause significant damage, and UVA is unaffected by ozone depletion. Herein, we show that the temperate ecosystems of southern South America have been subjected to increasingly high levels of ozone depletion during the last decade. We found that in the spring of 1997, despite frequent cloud cover, the passages of the ozone hole over Tierra del Fuego (55 degrees S) caused concomitant increases in solar UV and that the enhanced ground-level UV led to significant increases in DNA damage in the native plant Gunnera magellanica. The fluctuations in solar UV explained a large proportion of the variation in DNA damage (up to 68%), particularly when the solar UV was weighted for biological effectiveness according to action spectra that assume a sharp decline in quantum efficiency with increasing wavelength from the UVB into the UVA regions of the spectrum.

EM waves standards effectiveness.

There are many studies about the effect of electromagnetic radiations on living organisms. Most of these have been carried out with mice, rats, guinea pigs, rabbits, dogs, monkeys, etc; only a limited amount of data is based on effects observed directly on human beings, which were mostly exposed to EM waves because of professional reasons (Epidemiology). In this paper some of these studies have been gathered according to the affected system. At higher frequencies power density is the parameter to be taken into account, at lower frequencies electric and magnetic field are, but in the graph they were expressed in terms of equivalent power density as ExH when both field strength were given, or as E2/Z or H2 Z (where Z is the free space impedance), when only one of them was given. Standards from different countries to human exposure are presented, and finally an analysis of the effects compared with standards from the U.S.A. and U.S.S.R. is presented.