Spin-labeling study of phosphatidylcholine-cardiolipin binary mixtures

Electron paramagnetic resonance spectra of the spin-label probe 2,2,6,6-tetramethylpiperidinyl-1-oxy have been used to study the phase behavior of binary mixtures of different phosphatidylcholines (dipalmitoyl, distearoyl, and dioleoyl) with cardiolipin, using either calcium-free or calcium-containing cardiolipin (with a calcium:cardiolipin ratio of 1:2) samples. Results show that the nature of the fatty acid chains of the phosphatidylcholines (chain length and unsaturation) may influence the coexistence of different phases as well as does the nature of the cation linked to the cardiolipin.     

Multiple phase equilibria in binary mixtures of phospholipids

Approximate phase diagrams describing lateral phase separations are given for binary mixtures of dimyristoyl phosphatidylcholine with dipalmitoyl phosphatidylcholine, distearoyl phosphatidylcholine, and dipalmitoyl phosphatidylethanolamine. These diagrams are based in part on freeze-fracture electron microscopic data presented here, as well as other earlier spin-label, calorimetric, and X-ray data. These phase diagrams represent an improvement over previous studies in that both solid phases (P_β' and L_β') of the phosphatidylcholines are included. Further consideration is given to the problem of binary mixtures in which there are two P_β' phases that do not form a continuous range of solid solutions.     

Supramolecular interactions of solid human serum albumin with binary mixtures of solvent vapors.

Sorption isotherms of organic compounds on solid human serum albumin (HSA) from binary vapor mixtures were determined by gas chromatographic headspace analysis. The shape of sorption isotherms depends on molecular structure of studied sorbates. The 'active' compounds capable to sorb effectively on dry HSA increase the sorption of 'passive' compounds unable to be sorbed by dry HSA in absence of the third component. The critical hydration of HSA is required for sorption activation of 'passive' sorbates if water is taken as 'active' component. Ethanol and acetonitrile exhibit such activation effect without threshold. 'Passive' sorbates are able to produce cooperative activation effect on the sorption of 'active' component. Hydration history effect is observed for sorption on prehydrated HSA and HSA hydrated in situ. Obtained results were interpreted in terms of clathrate formation by 'passive' sorbate (substrate) and 'active' component inside the HSA (receptor) binding centers.     

Phase Equilibria in binary mixtures of dimyristoylphosphatidylcholine and cardiolipin

Paramagnetic resonance spectra of the spin-label 2,2,6,6-tetramethylpiperidinyl-l-oxy have been used to study phase separations in binary mixtures of dimyristoyl-phosphatidylcholine and cardiolipin. Two different samples of cardiolipin were used: (i) One sample contained calcium ions at a mole ratio of calcium:cardiolipin = 1:2; the experimental data support the view that cardiolipin is present in the bilayer membrane as calcium ion linked dimers, (CL)2 Ca2+. (ii) A calcium-free sodium cardiolipin sample yielded remarkable spin-label partition data that were quite different from those obtained in the presence of Ca2+. In both cases the spin-label data provide evidence for compound formation and for fluid-fluid immiscibility in the bilayer membrane.     

Perception of pungent odorants alone and in binary mixtures

The present investigation explores the stimulus–response(psychophysical) functions for total nasal perceived intensityfor two pungent odorants, formaldehyde and ammonia, presentedeither alone or with different backgrounds of the other irritant.Stimuli were comprised of four formaldehyde concentrations (1.0,3.5, 6.9 and 16.7 p.p.m.); four ammonia concentrations (210,776, 1172 and 1716 p.p.m.); and their 16 binary mixtures. Ammoniafunctions showed a consistent upward concavity. At low, mediumand high concentrations, the total perceived intensity of themixtures showed hypoadditivity, simple additivity and hyperadditivity,respectively. That is, the intensity of the corresponding mixtureswas significantly lower than, equal to, or greater than thesum of its components. The progressive involvement of pungency,aroused by common chemical sense stimulation, may be responsiblefor the increasing additivity observed. Simple additivity, andeven hyperadditivity, may characterize mixtures involving commonchemical stimulation.     

Phase equilibria in binary mixtures of phosphatidylcholine and cholesterol

The paramagnetic resonance spectra of two spin-labels, 2,2,6,6-tetramethylpiperadinyl-1-oxy and a head-group spin-labeled phosphatidylethanolamine (L-alpha-dipalmitoylphosphatidyl-N-ethanolamine), have been used to study solid-liquid and liquid-liquid phase separations in binary mixtures of dimyristoylphosphatidylcholine and cholesterol. A quantitative analysis of these resonance spectra supports the view that at temperatures below theta m, the chain-melting temperature of the phospholipid, and at cholesterol mole fractions Xc less than 0.2, these mixtures consist of two phases, a solid phase of essentially pure dimyristoylphosphatidylcholine and a fluid phase having a mole fraction of cholesterol equal to 0.2. The spin-label data also provide evidence for fluid-fluid immiscibility in the bilayer membrane at temperatures above the chain melting transition temperature of dimyristoylphosphatidylcholine.     

Properties of lipid-apolipoprotein association products. Complexes of human apo AI and binary phospholipid mixtures

The products resulting from the association of human apo A-I with binary phospholipid mixtures of dimyristoyl phosphatidylcholine (DMPC) and either dipalmitoyl (DPPC) or distearoyl (DSPC) phosphatidylcholine have been isolated and characterized. Effective lipid . protein complex formation was found to occur at the onset temperature for melting of the gel state, and equal incorporation of both lipid components of the binary mixture was observed. Two sizes of products were obtained, one containing 2 A-I molecules per complex and the other containing 3; the proportions of these two products depended upon the initial phospholipid/protein ratio employed. these two product species were found to be resolvable by density gradient centrifugation as well as gel filtration, reflecting substantial differences in composition as well as size. The ratio of DMPC to DPPC or DSPC was the same in the isolated complexes as in the initial mixture, suggesting that th protein does not associate preferentially with the fluid phase lipid, but with lipid domains in which the components are randomly distributed. Electron microscopy of recombinant particles containing a 2:1 ratio (w/w) of DSPC to DMPC revealed stacks of discs whose thickness was proportionately greater than for discs containing DMPC alone. Also of significance was the finding that recombinant discs containing 3 A-I molecules possessed a diameter approximately 1.5 times larger than recombinant discs containing 2 A-I molecules. These data are consistent with the model for the recombinant particles described by Tall et al. (Tall, A.R., Small, D.M., Deckelbau, R.J., and Shipley, G.G. (1977) J. Biol. Chem. 252; 4701-4711), in which the phospholipid is found as a circular bilayer, the thickness of which is dependent upon the length of the acyl chain, and around which the protein is distributed as an annulus.     

Dilatometric study of binary mixtures of phosphatidylcholines.

Volumes of lipid dispersions as a function of temperature have been measured for two different kinds of binary mixtures of lecithins, (1) DMPC and DSPC and (2) DMPC and DC20PC. Emphasis was placed on DMPC-rich compositions so as to resolve ambiguities regarding solid-phase immiscibility in DMPC-DSPC mixtures. Special attention has been paid to problems of equilibration in the low-temperature phase and to methods of mixing the lipids. We find that there is no solid-solid immiscibility in DMPC-DSPC mixtures, although this system is close to exhibiting such immiscibility, and that DMPC-DC20PC mixtures exhibit pronounced solid immiscibility.     

Odor intensity of binary mixtures of odorous compounds

This paper presents the MBO model for the perceived intensity of odour mixtures. This model is based on an equation previously reported by our team, intended to model the whole stimulus-response intensity curve of pure odorous compounds. The MBO model was applied to a set of published data, and compared to other published models. The results show a high modelling efficiency of the MBO model compared to other proposed equations, especially for binary mixtures exhibiting significant asymmetry of intensity for different ratios of the two components. Furthermore, the MBO model includes parameters specific to the respective effects of each component in the mixture, which may help to clarify the masking and synergy effects that are often sought in odour mixtures.     

Amylose conformational transitions in binary DMSO/water mixtures

The effects on amylose conformation of percentage water in dimethyl sulfoxide (DMSO)/water mixtures were measured by following changes in specific optical rotation, limiting viscosity number, and ¹³C-NMR chemical shifts. The temperature dependence of specific optical rotation showed differences in amylose conformation at four chosen ratios of dimethyl sulfoxide/water. An amylose conformational change was also deduced from ¹³C-NMR chemical shift data. Changes in limiting viscosity of amylose in different proportions of DMSO/water, and the effect of tetramethylurea on the specific rotation of amylose, indicate that intramolecular hydrogen bonding decreases with increased water content. 66.6% DMSO appears to be a crossover concentration, below which the helical conformation is progressively lost as water is added. When water content is over 60%, transition to a conformation which allows iodine complexation to take place is complete. A transition of amylose conformation from helix to loose helix to random coil with increasing water content was deduced from the experimental results.     

Crowding effects in binary mixtures of rod-like and spherical particles

Crowding effects relevant to the phase stability of binary mixtures of rod-like and spherical particles are investigated by means of Monte Carlo simulations in the isobaric NPT ensemble. The two types of particles are represented, respectively, by freely rotating hard spherocylinders of a moderate aspect ratio (L/sigma = 5) and hard spheres of the same diameter sigma. Molar fractions of spheres ranging xHS = 0.00-0.37 are considered with the aim of characterizing the crowding effects on the liquid crystal phases of the hard spherocylinder fluid induced by the spherical component as depleting agent. We find that the addition of the spherical crowder is beneficial for the stabilization of the layers of the rod-like particles characteristic of the smectic phase. On the contrary, the addition of spheres has a negative impact upon the stability of the nematic phase, where the rod-like particles tend to align collectively parallel to each other. Interestingly, the spheres tend to arrange forming rod-like clusters in the nematic phase and lamellar structures in the smectic phase, which is compensated by the entropy gained by the spherocylinder particles in each phase. The main results are in qualitative agreement with recent experimental and theoretical studies and serve to test the prediction of current equations of state for these types of binary mixtures.     

An alternative model of olfactory quantitative interaction in binary mixtures

The "vectorial model" proposed in 1973 by Berglund et al. certainlyconstitutes an important progress in the field of olfactoryquantitative interaction. An alternative model called "U model",based also upon perceived odorous intensity of a mixture asa function of perceived odorous intensity of the components,is here presented. The "U model" fits the experimental data of Cain and Drexler(1974) and of Cain (1975) slightly better than the "vectorialmodel". ^*Presented at the VIth International Symposium Olfaction andTaste, Gif-sur-Yvette, Paris, France, 15–17th July, 1977.     

Prediction models for the pleasantness of binary mixtures in olfaction

Whereas the rules underlying the perceived intensity of binary mixtures have been investigated, minimal efforts have been directed at elucidating the rules underlying the perceived pleasantness of such mixtures. To address this, 84 subjects ranked the pleasantness and intensity of 5 distinct binary mixtures (15 pairs, inter-stimulus interval = 4 s, inter-trial interval = 30 s, flow = 6 l/min, pulse = 2 s) constructed from different ratios (0:100%, 25:75%, 50:50%, 75:25%, and 100:0%, olfactometer-generated vapor phase). We found that in the majority of cases, the pleasantness of the mixture fell between the pleasantness values of its separated constituents and that it was strongly influenced by the relative intensities of the constituents. Based on these results, we proposed a prediction paradigm for the pleasantness of binary mixtures from the pleasantness of their separated constituents weighted by their respective perceived intensities. The uniqueness of the proposed paradigm is that it neither requires presetting an interaction constant between the mixture components nor require any factorization of the pleasantness weights. It does, nonetheless, require solid psychophysical data on the separated components at their different concentrations, and currently it can only explain the behavior of intermediate pleasantness of mixtures.     

Aggregation of biopharmaceuticals in human plasma and human serum

Analytical methods based on light microscopy, 90° light-scattering and surface plasmon resonance (SPR) allowed the characterization of aggregation that can occur when antibodies are mixed with human plasma. Light microscopy showed that aggregates formed when human plasma was mixed with 5% dextrose solutions of Herceptin^® (trastuzumab) or Avastin^® (bevacizumab) but not Remicade^® (infliximab). The aggregates in the plasma-Herceptin^®-5% dextrose solution were globular, size range 0.5–9 μm, with a mean diameter of 4 μm. The aggregates in the plasma-Avastin^®-5% dextrose samples had a mean size of 2 μm. No aggregation was observed when 0.9% NaCl solutions of Herceptin^®, Avastin^® and Remicade^® were mixed with human plasma. 90° light-scattering measurements showed that aggregates were still present 2.5 h after mixing Herceptin^® or Avastin^® with 5% dextrose-plasma solution. A SPR method was utilized to qualitatively describe the extent of interactions of surface-bound antibodies with undiluted human serum. Increased binding was observed in the case of Erbitux^® (cetuximab), whereas no binding was measured for Humira^® (adalimumab). The binding of sera components to 13 monoclonal antibodies was measured and correlated with known serum binding properties of the antibodies. The data presented in this paper provide analytical methods to study the intrinsic and buffer-dependent aggregation tendencies of therapeutic proteins when mixed with human plasma and serum.     

Perceptual interactions in odour mixtures: odour quality in binary mixtures of woody and fruity wine odorants

The qualitative perceptual interactions in three binary mixtures of wine odorants were studied: isoamyl acetate (fruity note)/whisky lactone (woody note), ethyl butyrate (fruity note)/whisky lactone (woody note) and ethyl butyrate (fruity note)/guaiacol (woody note). For each binary mixture, the perceived quality and intensity of 24 stimuli (four supra-threshold concentration levels of each of the two compounds and their 16 possible combinations) were evaluated in five replications by a trained panel of 13 subjects. The application of the Olsson predictive model for odour intensity and quality perception gave quite a good estimation of the evolution of single component identification in the mixture when the intensity proportion of unmixed components varied. However, this model was unable to account for the odour quality dominance in mixtures of iso-intense components. An alternative linear logistic model was proposed to study the qualitative dominance of the woody note in the three mixtures when the perceived intensities of each unmixed compound were equal.     

The similarity between odors and their binary mixtures in Drosophila

How are odor mixtures perceived? We take a behavioral approach toward this question, using associative odor-recognition experiments in Drosophila. We test how strongly flies avoid a binary mixture after punishment training with one of its constituent elements and how much, in turn, flies avoid an odor element if it had been a component of a previously punished binary mixture. A distinguishing feature of our approach is that we first adjust odors for task-relevant behavioral potency, that is, for equal learnability. Doing so, we find that 1) generalization between mixture and elements is symmetrical and partial, 2) elements are equally similar to all mixtures containing it and that 3) mixtures are equally similar to both their constituent elements. As boundary conditions for the applicability of these rules, we note that first, although variations in learnability are small and remain below statistical cut-off, these variations nevertheless correlate with the elements' perceptual "weight" in the mixture; thus, even small differences in learnability between the elements have the potential to feign mixture asymmetries. Second, the more distant the elements of a mixture are to each other in terms of their physicochemical properties, the more distant the flies regard the elements from the mixture. Thus, titrating for task-relevant behavioral potency and taking into account physicochemical relatedness of odors reveals rules of mixture perception that, maybe surprisingly, appear to be fairly simple.     

Induction of lateral phase separations in binary lipid mixtures by alcohol

It has previously been shown that alcohol has different effects on the gel to liquid-crystal phase transition of phosphatidylcholines (PC's) and phosphatidylethanolamines (PE's) [Rowe, E. S. (1985) Biochim. Biophys. Acta 813, 321-330]. In this investigation, the thermotropic properties of binary PE-PC mixtures were studied in the presence of ethanol in order to determine whether the differential interactions of alcohol with PC and PE would lead to lateral phase separations. Phase diagrams of the dilaurylphosphatidylethanolamine-dipalmitoylphosphatidylcholine [PE(12:0)-PC(16:0)] system were constructed in the presence and absence of ethanol. It was shown that lateral phase separations occur in the gel phase over a certain composition range in the presence of 100 mg/mL ethanol. In the absence of alcohol these two lipids are miscible in both the gel and liquid-crystal states. The data suggest that in the presence of ethanol these lateral phase separations involve the coexistence of regular bilayer gel and the fully interdigitated gel phase, which has previously been shown to occur in pure PC(16:0) under these conditions [Simon, S. A., & McIntosh, T. J. (1984) Biochim. Biophys. Acta 773, 169-172]. The biological implications of these findings are discussed.     

Miscibility of phosphatidylcholine binary mixtures in unilamellar vesicles: Phase equilibria

Summary Miscibility among phospholipids with different lipid chain-lengths or with different head groups has attracted a number of research efforts because of its significance in biological membrane structure and function. The general consensus about the miscibility of phosphatidylcholines with varying lipid chainlengths appears to be that binary mixtures of phospholipids with a difference of two carbon atoms in the lipid chain mix well at the main phase transition. Miscibility between phosphatidylcholines with differences of four carbon atoms appears to be inconclusive. Previous reports on the phase transition of binary phospholipid mixtures are concerned mainly with multilamellar vesicles and are mostly limited to the main transition. In the present study, unilamellar vesicles were used and miscibility in binary systems between dimyristoyl-, dipalmitoyl- and distearoyl-phosphatidylcholines at pretransition, as well as main transition temperatures was evaluated by constructing phase diagrams. Two methods were used to monitor the phase transitions: differential scanning microcalorimetry and optical absorbance methods. The optical method has the advantage that unilamellar vesicles of dilute phospholipid concentrations can be used. The liquidus and solidus phase boundaries were determined by the onset temperature of heating and cooling scans, respectively, because the completion temperature of a phase transition has no meaning in binary solutions. Dimyristoyl- and distearoyl-phosphatidylcholines. where the difference in the, lipid chain-length is four carbon atoms, mixed well even at pretransition temperature.     

Kinetics of biodegradation of binary and ternary mixtures of PAHs

The kinetics of biodegradation of mixtures of polycyclic aromatic hydrocarbons (PAHs) by Sphingomonas paucimobilis strain EPA505 were investigated. The investigation focused on three- and four-ring PAHs, specifically 2-methylphenanthrene, fluoranthene, and pyrene. Uptake rates in aerobic batch suspended cultivations were measured for the individual PAHs and their binary and ternary mixtures. It was observed that kinetics were influenced by the mixture composition and the kinetic properties of the components. A material balance equation containing the Monod model was numerically fitted to uptake data to determine extant kinetic parameters for the individual PAHs. Similarly, equations containing kinetic interaction models derived from enzyme kinetics were fitted to the uptake data obtained from experiments with binary and ternary mixtures. The investigation considered the following interaction types: no-interaction (Monod), pure competitive interaction, noncompetitive or mixed-type interaction, uncompetitive inhibition, and nonspecific interaction based on pure competition (SKIP). Model fit was evaluated based on probabilistic and statistical criteria and inferences were reached about underlying interaction mechanisms based on model fit. Mixture kinetics were most adequately simulated by the pure competitive interaction model with mutual substrate exclusivity. This model is fully predictive, relying only on parameters determined in the sole-PAH experiments. It was shown that for low percent inhibition values and with limited data, pure competitive interaction kinetics may not be evident, resembling no-interaction kinetics. This study is a reasonable starting point for understanding and modeling biodegradation of complex PAH mixtures in engineered and natural systems.