Molecular dynamics simulations of bilayers containing mixtures of sphingomyelin with cholesterol and phosphatidylcholine with cholesterol

We performed six molecular dynamics simulations: three on hydrated bilayers containing pure phospholipids and three on hydrated bilayers containing mixtures of these phospholipids with cholesterol. The phospholipids in our simulations were SSM (sphingomyelin containing a saturated 18:0 acyl chain), OSM (sphingomyelin with an unsaturated 18:1 acyl chain), and POPC (palmitoyloleoylphosphatidylcholine containing one saturated and one unsaturated chain). Data from our simulations were used to study systematically the effect of cholesterol on phospholipids that differed in their headgroup and tail composition. In addition to the structural analysis, we performed an energetic analysis and observed that energies of interaction between cholesterol and neighboring SM molecules are similar to the energies of interaction between cholesterol and POPC. We also observed that the interaction energy between cholesterol and neighboring lipids cannot be used for the determination of which lipids are involved in the creation of a complex.     

Raftlike mixtures of sphingomyelin and cholesterol investigated by solid-state 2H NMR spectroscopy

Sphingomyelin is a lipid that is abundant in the nervous systems of mammals, where it is associated with putative microdomains in cellular membranes and undergoes alterations due to aging or neurodegeneration. We investigated the effect of varying the concentration of cholesterol in binary and ternary mixtures with N-palmitoylsphingomyelin (PSM) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) using deuterium nuclear magnetic resonance ((2)H NMR) spectroscopy in both macroscopically aligned and unoriented multilamellar dispersions. In our experiments, we used PSM and POPC perdeuterated on the N-acyl and sn-1 acyl chains, respectively. By measuring solid-state (2)H NMR spectra of the two lipids separately in mixtures with the same compositions as a function of cholesterol mole fraction and temperature, we obtained clear evidence for the coexistence of two liquid-crystalline domains in distinct regions of the phase diagram. According to our analysis of the first moments M1 and the observed (2)H NMR spectra, one of the domains appears to be a liquid-ordered phase. We applied a mean-torque potential model as an additional tool to calculate the average hydrocarbon thickness, the area per lipid, and structural parameters such as chain extension and thermal expansion coefficient in order to further define the two coexisting phases. Our data imply that phase separation takes place in raftlike ternary PSM/POPC/cholesterol mixtures over a broad temperature range but vanishes at cholesterol concentrations equal to or greater than a mole fraction of 0.33. Cholesterol interacts preferentially with sphingomyelin only at smaller mole fractions, above which a homogeneous liquid-ordered phase is present. The reasons for these phase separation phenomena seem to be differences in the effects of cholesterol on the configurational order of the palmitoyl chains in PSM-d31 and POPC-d31 and a difference in the affinity of cholesterol for sphingomyelin observed at low temperatures. Hydrophobic matching explains the occurrence of raftlike domains in cellular membranes at intermediate cholesterol concentrations but not saturating amounts of cholesterol.     

Binary,ternary and quaternary complexes invloved in the systems pyridoxamine-glycine-imidazole with some bivalent metal ions

Equilibrium-based computer models using MINIQUAD-75 program were utilized to determine the stoichiometry and formation constants involved in the systems pyridoxamine(Pm)-glycine (Gly)-imidazole (lmd) with CO(II), Ni(II), Cu(II), Zn(II) and Cd(II) metal ions. The data were obtained from potentiometric pH titration of the various binary and ternary quaternary systems under physiological-like conditions (0.15 M NaNO₃-37°C). Various composition ratios of metal and ligands were used. The ligand concentrations did not exceed 4 times the concentration of metal ion in the binary systems and 4 times of the metal ions in ternary systems. In case of the quaternary systems only imidazole concentrations were two or four times the concentrations of metal ions keeping those of other ligands equal to that of metal ions. The stability constants of the quaternary species are discussed in terms of binary and ternary constants as are the effect of ring size on the stability of mixed ligand species. In addition, electrostatic as well as statistical effects also are mentioned and the biological implications of these model equilibria are described.     

Characterization of associates in ternary mixtures of amines, diprotic acid dyes and quaternary ammonium compounds in dichloromethane and in dichloromethane/water systems

Ternary mixtures of Bromocresol Green (BCGH), Benzethonium Chloride (BZ(+)Cl(-)), and Quinine (Q) in dichloromethane (CH(2)Cl(2) for ratios 1:>/=1:>/=1 (BCGH(2):BZ(+)Cl(-):Q) generate species BCGH(-)BZ(+), BZ(+)BCG(-)-H-Q) and BCG(2-) (BZ(+))(2) in chemical equilibrium; whose thermodynamic parameters are determined. A new method to study ternary mixtures in a non-polar solvent has been given and other amines (A) and quaternary ammonium compounds (QAC) instead of Q and BZ(+)Cl(-) have also been researched. Species BCGH(-)BZ(+), and BCG(2-)(BZ(+))(2) are ion associates of 1:1 and 1:2 (dye:BZ(+)Cl(-)) stoichiometry and species BZ(+)BCG(-)-H-Q presents a hydrogen bond, being of 1:1:1 (dye:BZ(+)Cl(-):Q) stoichiometry. The Vis-VU, IR and (1)H-NMR spectra of the associates suggest that they are in nature resonance hybrids. A new and fundamental equation which governs extraction of any 1:1:1 associate is deduced and checked experimentally, showing that its extraction depends on the high capacity of the amine to accept hydrogen bonds and the high extractability of the ammonium ion. Extraction of the 1:1:1 associate using different amines and ammonium ions is studied both, experimentally and by the new equation, checking that the 1:1:1 associate containing Q and BZ(+) is selectively extracted due to the fact that Q has a high hydrophobicity and high capacity to form hydrogen bonds and species BZ(+)Cl(-) has a high ion-associability. Selective extraction of this 1:1:1 associate is useful for quantitative determination in complex mixtures of ammonium ions of high ion associability as BZ(+)Cl(-).     

The effect of cholesterol on the adsorption of phenyltin compounds onto phosphatidylcholine and sphingomyelin liposome membranes

Organometallic compounds are widely spread in the human environment sometimes, causing a substantial health risk. Their amphiphilic character enables them to intercalate and penetrate cell membranes, potentially affecting various vital cell functions. Compound adsorption onto the membrane depends on the compound properties, as well as on the membrane composition and state. When adsorbing onto the lipidic surface, phenyltins localize at areas where lipid bilayer organization is compatible with compound spatial requirements. The lipid bilayer is a dynamic and laterally nonuniform structure with complex local and global architecture correlated with a variety of cell functions. The selective binding of a toxic compound to selected membrane areas may, therefore, interfere with some types of cellular process. We present experimental results concerning phenyltin adsorption onto the lipid bilayer surface measured with the fluorescent probe fluorescein‐PE. Model lipid bilayers were formed from lipid mixtures mimicking various plasma membrane regions. The adsorption of Ph₃SnCl and P₂SnCl₂ onto the phosphatidylcholine–cholesterol bilayer was qualitatively different from sphingomyelin–cholesterol. The results presented indicate that phenyltins are likely to accumulate in areas containing phosphatidylcholine, outside of lipid rafts. Copyright © 2004 John Wiley & Sons, Ltd.     

Phase equilibrium measurements of ternary systems formed by linoleic and linolenic acids in carbon dioxide/ethanol mixtures

This work reports phase equilibrium measurements for the ternary systems linoleic (acid + CO₂ + ethanol) and (linolenic acid + CO₂ + ethanol). The fatty acids present in the ternary systems were selected based on composition of banana peel oil extracted by supercritical CO₂ at 20 MPa and 313 K. The motivation of this research relies on the fact that these unsaturated fatty acids are recognized to play an important role in lowering blood pressure and serum cholesterol and because they are present in high concentrations in banana peel extract. Besides that, equilibrium data of these compounds are scarce in literature. The phase equilibrium experiments were performed using a high-pressure variable-volume view cell over the temperature range of (303 to 343) K and pressures up to 19 MPa. For both systems, only vapour–liquid phase transitions were visually recorded for all data measured.     

Studies on an ester-modified cationic amphiphile in aqueous systems: behavior of binary solutions and ternary mixtures with conventional surfactants

The aqueous behavior of an ester-modified cationic amphiphile with the molecular structure CH3CH2O(C=O)(CH2)6(C=O)O(CH2)8N+(CH3)3Br-, in the following referred to as A, has been investigated. Systems with A as the only solute, as well as different aqueous mixtures with conventional cationic surfactants, primarily dodecyltrimethylammonium bromide (DTAB), were included in the study. Isotropic solution samples were characterized using 1H NMR, 13C NMR, NMR diffusometry, and conductivity measurements, whereas liquid crystalline samples were investigated by optical polarization microscopy and small-angle X-ray diffraction. The results are compared to the behavior of the binary system of DTAB and water. A does not exhibit a typical surfactant behavior. When it is present as the only solute in a binary aqueous system, it forms neither conventional micelles nor liquid crystalline phases. However, there is clear evidence that it assembles with lower cooperativity into loose clusters at concentrations above 25-30 mM. When A is mixed with DTAB in solution, the two amphiphiles form mixed assemblies, the structure of which varies with the total amphiphile concentration. In concentrated mixtures with alkyltrimethylammonium surfactants, A can participate in hexagonal liquid crystalline phases even when it constitutes a significant fraction of the total amphiphile content.     

Reassessment of the binary, ternary, and quaternary interactions in mixed electrolytes from thermodynamic quantities: The systems with uncommon ions containing hydrophobic character

Accurate estimates of the binary, ternary, and quaternary interactions in aqueous ionic mixtures with uncommon ions with hydrophobic character are presented. For this purpose, the values of the excess Gibbs free energy of mixing, Delta m G(E), obtained from our earlier isopiestic osmotic coefficients (Kumar, A. J. Phys. Chem. B2003, 107, 2808) for the mixtures of NaCl with four guanidinium (Gn+) salts-CH3COOGn, GnNO3, GnClO4, and Gn2SO4-are analyzed with the help of the method developed by Leifer and Wigent. The methodology of Leifer and Wigent is based on the equations of Scatchard-Rush-Johnson and Friedman's cluster integral expansion theory. The Scatchard-Rush-Johnson theory explicitly considers the quaternary and higher-order ionic interactions in the mixtures as compared to the specific ion interaction theory of Pitzer, which accounts for binary and ternary interactions only. The contributions due to binary, ternary, and quaternary interaction terms to total Delta m G(E) are estimated and discussed critically. Also, the interaction between the same two cations, for example, Gn+ - Gn+, is estimated and found significant, which otherwise cannot be obtained by the use of Pitzer's theory. The information obtained from the analysis of Delta(m)G(E) is also supported by the newly measured excess volumes of mixing, Delta m V(E), at 298.15 K. The individual contributions of the binary, ternary, and quaternary interaction terms to total Delta m V(E) are described. The binary, ternary, and quaternary interaction terms for both Delta m G(E) and Delta m V(E) are analyzed in terms of Friedman's cluster integral expansion theory.     

Some selected problems of nonaqueous microemulsions — phase diagrams and variation of the composition in the quaternary and ternary systems

A variety of quaternary and ternary systems of the type of dodecane/aliphatic alcohols/Na-dodecyl sulphate/water or a water-replacing component (formamide, ethylene glycol, propylene glycol, propylene carbonate, dimethylsulfoxide, acetonitrile) were subjected to a comparative analysis of microemulsification. The compositions of the systems of the type of oil/surfactant/cosurfactant/water or nonaqueous liquid were changed with respect to surfactant (Na-dodecyl sulfate and triton X 114) concentration, cosurfactant (homologous aliphatic alcohols) concentration and mixing ratio of water and water-replacing component. The appertaining phase diagrams were plotted and compared with those of aqueous systems.The experimental results suggest that the phase regions, which are designated as nonaqueous microemulsions, probably represent, not only microemulsions in the usual sense, but a separate kind of mixed phase whose microstructure is examined by special studies. Analogously to aqueous microemulsion systems, transparency and spontaneous formation of homogeneous multicomponent systems cannot serve as the sole criteria for waterless microemulsion formation. But they are important guiding properties of microemulsion formation in multicomponent systems.The variety of components involved in the chemical composition and the current insufficient knowledge do not permit to generally decide whether nonaqueous systems can be assigned to microemulsions or to molecular solutions. For clarifying this problem a detailed examination of the miscibility behavior, especially that of binary systems as a function of temperature, and the role that surfactant and cosurfactant play in the formation of homogeneous systems will be necessary.     

The extraction of aromatic amino acids with binary and ternary mixtures of hydrophilic solvents

The extraction of tyrosine and phenylalanine with binary and ternary mixtures of hydrophilic solvents from aqueous salt solutions was studied, and several tendencies were observed. Simplex-lattice planning of experiment was used for the optimization of the composition of solvent mixtures. It was shown that the extraction systems developed could be employed for the almost complete extraction of tyrosine and phenylalanine from aqueous solutions.     

Thermal properties of ternary systems: The calculation of the standard enthalpy of solution for compounds in binary mixtures

An equation free of fitting parameters is proposed for calculating the standard heats of solution for compounds in nonaqueous binary mixtures. The parameters of the equation are the standard heats of solution of a compound in the components of the mixed solvent. Nonlinear ΔH ⁰(x) trends are reconstructed for solutions of water in i-PrOH + MeOH and MeCN + MeOH, t-BuOH in MeCN + MeOH, squalane in CHCl₃ + CCl₄ and C₆H₆ + CHCl₃, and hexadecane in MeOH + i-Pr₂O and in mixtures of butyl acetate, ethyl acetate, and 1,4-dioxane with 1-octanol. The standard heats of solutions are calculated for water in alcohol + alcohol, alcohol + aprotic solvent, and aprotic solvent + aprotic solvent mixtures     

Surfactant effects of chlorpromazine and imipramine on lipid bilayers containing sphingomyelin and cholesterol

The surface-active drugs chlorpromazine (CPZ) and imipramine (IP) have been tested on large unilamellar vesicles composed of phosphatidylcholine (PC), sphingomyelin (SM), and cholesterol (Ch) in different proportions. The well-characterized nonionic detergent Triton X-100 (TX) has also been used in parallel experiments. Leakage of vesicular aqueous contents and bilayer solubilization have been measured for each surfactant molecule and vesicle composition. All three surface-active molecules behave in a qualitatively similar way, irrespective of bilayer composition: they induce leakage at concentrations well below their critical micellar concentrations (cmc) and solubilization near the cmc. In these events, the potency of the three surfactants under study increases with decreasing cmc, in the order IP相似文献   

Ordering effects of cholesterol on sphingomyelin monolayers investigated by high-resolution broadband sum-frequency generation vibrational spectroscopy

After adding cholesterol, the sphingosine backbones (red) of the three nature SMs become more ordered, and the N-linked acyl chain (blue) remains unaltered.     

Interactions of Triton X-100 with sphingomyelin and phosphatidylcholine monolayers: influence of the cholesterol content

The presence of microdomains, called lipid rafts, in biological membranes is usually explained by lateral segregation between specific lipids and proteins. These rafts present similarities with the membrane domains isolated by their non-ionic detergent-resistance at 4 degrees C. They are enriched in sphingomyelin and cholesterol as compared with the outer leaflet of eukaryotic cell membranes. To understand the role played by the lipids enriched in rafts in their resistance to solubilization by detergents, the interactions between these lipids and the non-ionic detergent Triton X-100 were studied by using different lipid monolayers at the air-water interface. The influence of Triton X-100 on the Langmuir isotherms (i.e. surface pressure/area isotherms) of monolayers containing sphingomyelin and cholesterol at different mole ratios was analyzed and the results were compared with the influence of Triton X-100 on monolayers containing a phosphatidylcholine bearing a saturated and an unsaturated fatty acid (i.e. palmitoyloleylphosphatidylcholine) and cholesterol. This phosphatidylcholine was chosen since the phosphatidylcholines present in rafts isolated from bovine kidney could contain about 50% of saturated fatty acids. Triton X-100 induces an increase in the condensing effect observed as compared with ideal mixture of phospholipid/cholesterol. Triton X-100-induced changes in the morphology of the monolayers were visualized by Brewster angle microscopy, which confirmed the differences of behavior observed by analyzing the isotherms.     

Separation of ternary mixtures by pseudo-simulated moving bed chromatography.

Modeling and simulation are presented of the separation of a ternary mixture by pseudo-simulated moving bed chromatography, according to the JO process of Japan Organo Co. The process cycle is divided into two steps. In step 1, feed and eluent streams are introduced into the system, equivalent to a series of preparative chromatographic columns, and the intermediate component is produced. In step 2, similar to a simulated moving bed (SMB), there is no feed and the less adsorbed species is collected in the raffinate while the more retained species is collected in the extract; this step is described by an equivalent TMB model. A parametric study is presented in order to analyze the sensitivity of the model to variations on the duration of step 1 and step 2 and on the intraparticle mass transfer coefficient.     

Drug-excipient compatibility studies in binary and ternary mixtures by physico-chemical techniques

This work is part of a systematic study undertaken to find and optimize a general method of detecting the drug-excipient interactions, with the aim of predicting rapidly and inexpensively the long term stability of a pharmaceutical product and speed up its marketing. Here, in particular, the compatibility of haloperidol with several excipients (PVP, magnesium stearate and α-lactose) in binary and ternary mixtures, both as prepared and ball-milled, has been assessed by thermal methods, electron microscopy, IR spectroscopy and X-ray diffraction. The differences between the experimental behaviour of the systems and that expected as weighted average of similarly treated pure components are interaction indicators. The DSC has proven to be, among the selected analytical techniques, the most sensitive and specific in assessing the compatibility. A strong interaction has been observed between PVP and haloperidol. It is favoured by the mechanical stress and is more evident in the composition 20:80. On the contrary, α-lactose and magnesium stearate were found to be compatible with the drug.     

Modification of the NRTL model for ternary and quaternary liquid-liquid equilibrium calculations

The original NRTL model is modified for the correlation of ternary liquid-liquid equilibria. The ternary expression of the modified NRTL model includes three additional ternary parameters and the ternary terms vanish when a ternary system degenerates to a binary. The ability of the modified NRTL model has been evaluated in the calculations of ternary vapor-liquid-liquid equilibria and quaternary liquid-liquid equilibria.     

Cholesterol and phytosterols effect on sphingomyelin/phosphatidylcholine model membranes--thermodynamic analysis of the interactions in ternary monolayers

In this work thermodynamic analysis of the interactions between lipids in ternary sphingomyelin/DPPC/sterol Langmuir films were performed to compare the effect of cholesterol, beta-sitosterol and stigmasterol on a model membrane. The condensing effect of the respective sterols and the interactions between molecules in ternary mixtures were analyzed on the basis of the excess area per molecule and the excess free energy of mixing values. The stability of the mixed monolayers was verified with the free energy of mixing values. The conclusions on the ordering effect of sterols were drawn from the analysis of the compression modulus values. It was found that the stoichiometry of the mixed films of the highest thermodynamic stability and of the strongest interactions is the same for all the sterols investigated. The results obtained prove that the mammalian sterol induces the strongest contraction of the area and reveals the strongest stabilizing and ordering effect among the investigated sterol. Stigmasterol was found to condense a model membrane in a weaker extent as compared to beta-sitosterol, however, the differences in ordering properties of both phytosterols are less pronounced. The magnitude of the influence of the investigated sterols on a model membrane was thoroughly discussed from the point of view of the structure of their side chain, which determines the geometry of a sterol molecule.