The adsorption of phloretin to lipid monolayers and bilayers cannot be explained by langmuir adsorption isotherms alone

Phloretin and its analogs adsorb to the surfaces of lipid monolayers and bilayers and decrease the dipole potential. This reduces the conductance for anions and increases that for cations on artificial and biological membranes. The relationship between the change in the dipole potential and the aqueous concentration of phloretin has been explained previously by a Langmuir adsorption isotherm and a weak and therefore negligible contribution of the dipole-dipole interactions in the lipid surface. We demonstrate here that the Langmuir adsorption isotherm alone is not able to properly describe the effects of dipole molecule binding to lipid surfaces--we found significant deviations between experimental data and the fit with the Langmuir adsorption isotherm. We present here an alternative theoretical treatment that takes into account the strong interaction between membrane (monolayer) dipole field and the dipole moment of the adsorbed molecule. This treatment provides a much better fit of the experimental results derived from the measurements of surface potentials of lipid monolayers in the presence of phloretin. Similarly, the theory provides a much better fit of the phloretin-induced changes in the dipole potential of lipid bilayers, as assessed by the transport kinetics of the lipophilic ion dipicrylamine.     

Mechanical and biological properties of two types of bioactive bone cements containing MgO-CaO-SiO2-P2O5-CaF2 glass and glass-ceramic powder

The electrostatic steering of charged ligands toward the active site of Torpedo californica acetylcholinesterase is investigated by Brownian dynamics simulations of wild type enzyme and several mutated forms, in which some normally charged residues are neutralized. The simulations reveal that the total ligand influx through a surface of 42 A radius centered in the enzyme monomer and separated from the protein surface by 1-14 A is not significantly influenced by electrostatic interactions. Electrostatic effects are visible for encounters with a surface of 32 A radius, which is partially hidden inside the protein, but mostly within the solvent. A clear accumulation of encounter events for that sphere is observed in the area directly above the entrance to the active site gorge. In this area, the encounter events are increased by 40% compared to the case of a neutral ligand. However, the differences among the encounter rates for the various mutants considered here are not pronounced, all rate constants being within +/- 10% of the average value. The enzyme charge distribution becomes more important as the charged ligand moves toward the bottom of the gorge, where the active site is located. We show that neither the enzyme's total charge, nor its dipole moment, fully account for the electrostatic steering of ligand to the active site. Higher moments of the enzyme's charge distribution are also important. However, for a series of mutations for which the direction of the enzyme dipole moment is constant within a few degrees, one observes a gradual decrease in the diffusional encounter rate constant with the number of neutralized residues. On the other hand, for other mutants that change the direction of the dipole moment from that of the wild type, the calculated encounter rate constants can be very close to that of the wild type. The present work yields two new insights to the kinetics of acetylcholinesterase. First, evolution appears to have built a redundant electrostatic steering capability into this important enzyme through the overall distribution of its thousands of partially charged atoms. And second, roughly half of the rate enhancement due to electrostatics arises from steering of the substrate outside the enzyme; the other half of the rate enhancement arises from improved trapping of the substrate after it has entered the gorge. The computational results reproduce qualitatively, and help to rationalize, many surprising experimental results obtained recently for human acetylcholinesterase.     

Electrostatics and electrodynamics of bacteriorhodopsin

The stationary electric dichroism of bacteriorhodopsin is in qualitative, but not quantitative, agreement with the orientation function for disks having a permanent dipole directed perpendicular to the plane and an induced dipole in the plane. Fits of the orientation function to data measured at low field strengths demonstrate: an increase of the permanent dipole moment mu with the square of the disk radius r2, whereas the polarizability alpha increases with r4; the ionic strength dependence is small for mu and clearly stronger for alpha; the permanent dipole moment is 4x10(6) D at r = 0.5 micron. According to the risetime constants, the induced dipole does not saturate and increases to 4x10(8) D at 40 kV/cm and r = 0.5 micron. The data indicate that the permanent dipole is not of some interfacial character but is due to a real assymetry of the charge distribution. The experimental dipole moment per protein monomer is approximately 55 D, whereas calculations based on the structure of Grigorieff et al. (Grigorieff, N., T.A. Ceska, K.H. Downing, J.M. Baldwin, and R. Henderson. 1996. Electron-crystallographic refinement of the structure of bacteriorhodopsin. J. Mol. Biol. 259:393-421) provide a dipole moment of approximately 570 D. The difference is probably due to a nonsymmetric distribution of charged lipid residues. It is concluded that experimental dipole moments reflect the mu-potential at the plane of shear for rotational diffusion, in analogy to the sigma-potential used for translational diffusion. It is suggested that the permanent dipole of bacteriorhodopsin supports proton transport by attraction of protons inside and repulsion of protons outside of the cell. Dichroism rise curves at field strengths between E = 150 and 800 V/cm reveal an exponential component with time constants tau 3r in the range between 1 and 40 ms, which is not found in Brownian dynamics simulations on a disk structure using hydrodynamic and electric parameters characteristic of bacteriorhodopsin disks. The experimental data suggest that this process reflects a cooperative change of the bacteriorhodopsin structure, which is induced already at a remarkably low field strength of approximately 150 V/cm.     

Transfer RNA mimicry in a new group of positive-strand RNA plant viruses, the furoviruses: differential aminoacylation between the RNA components of one genome

The effects of the organic calcium channel blocker verapamil and the beta-receptor blocker propranolol on dipole (phi(d)) and surface (phi(s)) potentials of bilayer lipid membranes were studied. The boundary potentials (phi(b)= phi(d) + phi(s)) of black lipid membranes, monitored by conductance measurements in the presence of nonactin and by capacitive current measurements were compared with phi(s) calculated from the electrophoretic mobility of lipid vesicles. It was shown that the increase of boundary potential, induced by the adsorption of the positively charged propranolol, was caused solely by an increase in surface potential. Although phi(s) also increases due to the adsorption of verapamil, phi(b) diminishes. A sharp decrease of the dipole potential was shown to be responsible for this effect. From Langmuir adsorption isotherm the dissociation constant Kd of verapamil was estimated. The uncharged form of verapamil (Kd=(0.061+/-0.01) mM at pH 10.5) has a tenfold higher affinity to a neutral bilayer membrane than the positively charged form. The alteration of membrane dipole potential due to verapamil adsorption may have important implications for both membrane translocation and partitioning of small or hydrophobic ions and charged groups of membrane proteins.     

Interaction of rabbit C-reactive protein with phospholipid monolayers studied by microfluorescence film balance with an externally applied electric field

C-reactive protein (CRP) is one of the most characteristic acute-phase proteins in humans and many other animals. It binds to phosphorylcholine in a calcium-dependent manner. In addition, CRP activates the complement systems via the classical pathway. The interaction between rabbit CRP (rCRP) and model biological membrane is studied using dimyristoylphosphatidylethanolamine and dipalmitoylphosphatidylcholine monolayers. Observations with fluorescence microscopy indicate that rCRP is more likely to be incorporated in the liquid phase of monolayers. Such incorporation does not depend on the presence of calcium and is not inhibited by phosphocholine. The area occupied by the protein when incorporated into the monolayer was estimated. The dipole moment density of the protein crossing the air/water interface was measured by applying an external electric field. Our results indicate that calcium binding leads to a conformational change in CPR, which might modify the orientation of CRP in the monolayer. In addition, a negative charge or negative difference in dipole moment density facilitates the incorporation of CPR into the monolayer.     

Imagining projective transformations: aligned orientations in spatial organization

Four experiments were conducted to investigate whether variations in orientation that profoundly affect the ability to imagine rotations also affect the ability to imagine projective transformations. For a basic rectilinear object and the three simpler Platonic Solids, imagining projective transformations (e.g., the casting of a shadow) was quite successful when the objects were aligned with the direction of projection. For the solids, this alignment occurred when the objects were generalized cylinders about axes aligned with the projection. As the objects were made more oblique to the projection, performance deteriorated markedly. When the objects were moderately aligned with the projection, performance depended on the orientation of the object and the orientation of the projection to the environment. We suggest that the imagination of projection and of rotation is a type of problem solving in which spatial structures are organized in relation to initially given properties of the objects and transformations. When there is alignment among the various structural components, this process of imagination works efficiently. Without such alignment, nonexperts often fail. We suggest that aligned (i.e., parallel and perpendicular) orientations are effective in spatial imagination because they are categorically distinct and singular, and they provide a critical form of redundancy.     

Specific Adsorption of Chromium Species on Kaolinite Surface

Specific adsorption of hydrolyzed chromium species on kaolinite surface was studied. Electrokinetic behavior of kaolinite particles in aqueous solutions in the presence of CrCl₃ showed that the hydrolyzed chromium species can adsorb on kaolinite surfaces and reverse its charge. This adsorption reached the maximum near the precipitation pH (ppt) of chromium hydroxide. The adsorption followed Langmuir isotherm and was irreversible. It might be resulted from hydrogen bonding between the hydrolyzed metal species and kaolinite surfaces. A novel chemical adsorption model was proposed between kaolinite surfaces and the hydrolyzed multivalent metal species.     

Parallel processing of objects in a naming task.

The authors investigated whether speakers who named several objects processed them sequentially or in parallel. Speakers named object triplets, arranged in a triangle, in the order left, right, and bottom object. The left object was easy or difficult to identify and name. During the saccade from the left to the right object, the right object shown at trial onset (the interloper) was replaced by a new object (the target), which the speakers named. Interloper and target were identical or unrelated objects, or they were conceptually unrelated objects with the same name (e.g., bat [animal] and [baseball] bat). The mean duration of the gazes to the target was shorter when interloper and target were identical or had the same name than when they were unrelated. The facilitatory effects of identical and homophonous interlopers were significantly larger when the left object was easy to process than when it was difficult to process. This interaction demonstrates that the speakers processed the left and right objects in parallel. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The short extracellular domain of the T cell receptor zeta chain is involved in assembly and signal transduction

The zeta chain is required in the TCR complex to guarantee its surface expression and function. However, an understanding of the interaction(s) between the zeta chain and the other proteins in the TCR/CD3 has not yet been achieved. In this report, we attempt to assign a functional role to the short extracellular (EC) domain of the zeta chain by studying its unique positive charge, a lysine at position 9, because of its interesting location to the interchain disulphide bond of the zeta chain homodimer. We show that amino acid exchanges of lysine 9 to glycine, serine, cysteine or asparagine generate TCR complexes which are clearly defective in antigenic signalling. Interestingly, the non-conservative point mutations were segregating TCR complex signalling pathways. However, lysine 9 is not critical for TCR complex surface expression unless the positively charged lysine is exchanged for the negatively charged amino acid aspartic acid. The zeta chain mutant bearing a lysine to cysteine exchange is the sole mutant to be inefficiently co-precipitated with the TCR/CD3 complex suggesting a loose interaction of the zeta chain within the TCR complex.     

The "Swedish" mutation of the amyloid precursor protein (APPswe) dissociates components of object-location memory in aged Tg2576 mice.

Aged Tg2576 mice show abnormalities in hippocampal morphology and physiology and display behavioral deficits in spatial navigation tasks consonant with a deficit in the functional properties of the hippocampus. However, the nature of the spatial representations disrupted by the "Swedish" mutation of the amyloid precursor protein (APPswe) is unclear. In an effort to characterize the memory deficits in Tg2576 mice, the spontaneous object exploration paradigm was used to interrogate spatial and object memory in mice. With object arrays of comparable size, 16-month-old Tg2576 mice showed a normal object familiarity/novelty effect but impaired memory for the location of objects when 2 objects exchanged locations (topological transformation; Experiment 1). In contrast, Tg2576 mice showed preferential exploration of familiar objects when they were moved to previously unoccupied locations (Experiment 2), irrespective of whether the transformation altered the metric properties of the object array (Experiments 3). These results suggest that Tg2576 mice are able to form representations of the identity of objects and a memory of the spatial organization of objects in an arena. In contrast, conjunctive memory for specific object-location associations is severely impaired in aged Tg2576 mice. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Vasculopathy and insulin resistance in the JCR:LA-cp rat

The fluorescent dye FM1-43 labels nerve terminals in an activity-dependent fashion and has been found increasingly useful in exploring the exo- and endocytosis of synaptic vesicles and other cells by fluorescence methods. The dye distributes between the aqueous phase and the lipid membrane but the physical-chemical parameters characterizing the adsorption/partition equilibrium have not yet been determined. Fluorescence spectroscopy alone is not sufficient for a detailed elucidation of the adsorption mechanism since the method can be applied only in a rather narrow low-concentration window. In addition to fluorescence spectroscopy, we have therefore employed high sensitivity isothermal titration calorimetry (ITC) and deuterium magnetic resonance (2H-NMR). ITC allows the measurement of the adsorption isotherm up to 100 microM dye concentration whereas 2H-NMR provides information on the location of the dye with respect to the plane of the membrane. Dye adsorption/partition isotherms were measured for neutral and negatively-charged phospholipid vesicles. A non-linear dependence between the extent of adsorption and the free dye concentration was observed. Though the adsorption was mainly driven by the insertion of the non-polar part of the dye into the hydrophobic membrane interior, the adsorption equilibrium was further modulated by an electrostatic attraction/repulsion interaction of the cationic dye (z=+2) with the membrane surface. The Gouy-Chapman theory was employed to separate electrostatic and hydrophobic effects. After correcting for electrostatic effects, the dye-membrane interaction could be described by a simple partition equilibrium (Xb=Kcdye) with a partition constant of 103-104 M-1, a partition enthalpy of DeltaH=-2.0 kcal/mol and a free energy of binding of DeltaG=-7.8 kcal/mol. The insertion of FM1-43 into lipid membranes at room temperature is thus an entropy-driven reaction following the classical hydrophobic effect. Deuterium nuclear magnetic resonance provided insight into the structural changes of the lipid bilayer induced by the insertion of FM1-43. The dye disturbed the packing of the fatty acyl chains and decreased the fatty acyl chain order. FM1-43 also induced a conformational change in the phosphocholine headgroup. The -P-N+ dipole was parallel to the membrane surface in the absence of dye and was rotated with its positive end towards the water phase upon dye insertion. The extent of rotation was, however, much smaller than that induced by other cationic molecules of similar charge, suggesting an alignment of FM1-43 such that the POPC phosphate group is sandwiched by the two quaternary FM1-43 ammonium groups. In such an arrangement the two cationic charges counteract each other in a rotation of the -P-N+ dipole.     

Effect of rare-earth doping on adsorption of carbon atom on ferrum surface and in ferrum subsurface: A first-principles study

The adsorption of carbon atom on Fe surface and in Fe subsurface with and without rare earth (La and Ce) substitution in the surface layer and subsurface layer was studied by first-principles calculations. The carbon atom is predicted to adsorb at hollow and long bridge site on Fe(100) and Fe(110), respectively. However, the carbon atom shifts to occupy preferentially hollow site on both Fe(100) and Fe(110) with rare earth atom doping at surface layer. The lower adsorption energies involved with stronger adsorption abilities were obtained for carbon atoms on Fe surface with rare earth doping at surface layer, which was determined by the electronic structure of the surface atoms. The La atom was pulled out the surface after carbon adsorption due to strong interaction of La–C, which is consistent with the more charge transfer. In the subsurface region, the carbon atom prefers to occupy at octahedral site with rare earth doping at surface layer in Fe slab. These strong adsorption energies of the carbon atoms on Fe surface and in Fe subsurface with rare earth pose relevant insights into the interaction between carbon and rare earth, which helps to understanding the influence mechanism of rare earth in carburizing.     

Light chain-dependent myosin structural dynamics in solution investigated by transient electrical birefringence

The technique of transient electrical birefringence was used to compare some of the electric and structural dynamic properties of myosin subfragment 1 (S1(elc, rlc)), which has both the essential and regulatory light chains bound, to S1(elc), which has only an essential light chain. The rates of rotational Brownian motion indicate that S1(elc, rlc) is larger, as expected. The permanent electric dipole moment of S1(elc, rlc) is also larger, indicating that the regulatory light chain portion of S1(elc, rlc) has a dipole moment and that it is aligned head-to-tail with the dipole moment of the S1(elc) portion. The permanent electric dipoles decrease with increasing ionic strength, apparently because of ion binding to surface charges. Both S1(elc, rlc) and S1(elc) have intrinsic segmental flexibility, as detected by the ability to selectively align segments with a brief weak electric field. However, unlike S1(elc), which can be structurally distorted by the action of a brief strong electric field, S1(elc, rlc) is stiffer and cannot be distorted by fields as high as 7800 V/cm applied to its approximately 8000 D permanent electric dipole moment. The S1 . MgADP . Pi analog S1 . MgADP . Vi is smaller than S1 . MgADP, for both S1(elc, rlc) and S1(elc). Interestingly, the smaller, stiffer S1(elc, rlc) . MgADP . Vi complex retains intrinsic segmental flexibility. These results are discussed within a framework of current hypotheses of force-producing mechanisms that involve S1 segmental motion and/or the loss of cross-bridge flexibility during force production.     

Structural requirements for the cytotoxicity of the N-terminal region of HIV type 1 Nef

We have found that the hemolytic and cytotoxic activities of myristoylated Nef N-terminal peptides require a net positive charge in the first seven amino residues of the sequence. The activities are considerably less dependent on the secondary structure of the peptides. Film balance studies showed that both active and inactive peptides interacted with neutral phospholipid monolayers, suggesting that binding to neutral lipids was not a sufficient condition for lytic activity. It was also found that nonmyristoylated N-terminal peptide did not interact to the same extent with the monolayer, indicating that myristoylation was essential for lipid interaction. It is considered that the positively charged residues of the proximate N terminus of Nef interact with acidic lipids of biological membranes, reinforcing the weak membrane-targeting properties of the myristyl chain. Parallels are drawn between this mode of interaction with membranes and that of members of the Src family of proteins, which are also myristoylated and have positively charged residues in their proximate N termini. In particular, these proteins and Nef also have serine residues in their proximal N-terminal regions, which when phosphorylated could neutralize the positive charge and thus provide a mechanism for modulating membrane interaction.     

Position-specific Asp-Lys pairing can affect signal sequence function and membrane protein topology

Positively charged amino acids are major determinants of the topology of bacterial inner membrane proteins, whereas negatively charged residues by themselves have little or no influence on the transmembrane orientation. Further, positively charged amino acids can very efficiently block the function of signal sequences when placed immediately downstream, while negatively charged residues are much less potent also in this regard. Here, we show that a negatively charged aspartic acid situated close to a positively charged lysine can attenuate both of these effects in a position-specific manner, suggesting that intra- or intermolecular charge pairing can modulate the interactions between positively charged residues in the nascent chain and parts of the secretory machinery or membrane phospholipids. These observations further underscore the importance of charged amino acids during protein translocation and membrane protein assembly.     

The preference of tryptophan for membrane interfaces

One of the ubiquitous features of membrane proteins is the preference of tryptophan and tyrosine residues for membrane surfaces that presumably arises from enhanced stability due to distinct interfacial interactions. The physical basis for this preference is widely believed to arise from amphipathic interactions related to imino group hydrogen bonding and/or dipole interactions. We have examined these and other possibilities for tryptophan's interfacial preference by using 1H magic angle spinning (MAS) chemical shift measurements, two-dimensional (2D) nuclear Overhauser effect spectroscopy (2D-NOESY) 1H MAS NMR, and solid state 2H NMR to study the interactions of four tryptophan analogues with phosphatidylcholine membranes. We find that the analogues reside in the vicinity of the glycerol group where they all cause similar modest changes in acyl chain organization and that hydrocarbon penetration was not increased by reduction of hydrogen bonding or electric dipole interaction ability. These observations rule out simple amphipathic or dipolar interactions as the physical basis for the interfacial preference. More likely, the preference is dominated by tryptophan's flat rigid shape that limits access to the hydrocarbon core and its pi electronic structure and associated quadrupolar moment (aromaticity) that favor residing in the electrostatically complex interface environment.     

Characterization of rash with indinavir in a national patient cohort

FTIR absorption spectra of the fundamental bands of LiF and LiCl were measured with an apodized resolution of 0.01 cm-1. The Herman-Wallis analysis of these spectra led to the determination of the ratio of the dipole derivative to the permanent dipole moment, [μe/(dμ/dr)ere], for each molecule. The vibrational dependence of the dipole moment for each molecule has been already reported and these data were reanalyzed to determine the dipole moment function independently. The results from the Herman-Wallis analysis were compared with these results, and good agreement was found. This demonstrates that the Herman-Wallis analysis can produce reliable relative values of dipole moment functions. Copyright 1999 Academic Press.     

Adsorption of Negatively Charged Azo Dyes onto Surfactant-Modified Sepiolite

The adsorption of reactive dyes on sepiolite has been investigated in a series of batch adsorption experiments. Three reactive dyes (Everzol Black B, Everzol Red 3BS, Everzol Yellow 3RS H/C) were used in bottle adsorption studies. While no significant adsorption capacity was obtained for natural sepiolite, high-adsorptive capacities were observed upon using sepiolite modified with quaternary amines. The electrokinetic measurements along with calculations using the cross-sectional area reveal that the quaternary amines adsorb close to bilayer coverage. A mechanism involving electrostatic attraction of the anionic groups of dye molecules onto oppositely charged amine-modified sepiolite surface is proposed to be responsible for the uptake of dyes. The adsorption data were fitted to the Langmuir isotherm. It is found that the modified sepiolite yields adsorption capacities (qe) of 169, 120, and 108 mg/g for Yellow, Black, and Red, respectively. These results are comparable to the adsorption capacity of the same dyes onto activated carbon.     

Spontaneous representation of number in cotton-top tamarins ( Saguinus oedipus).

The "violation of expectancy" looking-time methodology has proven a powerful tool for exploring prelinguistic mental representations in human infants as well as in nonhuman primates. Four studies applying this methodology to the question of spontaneous number representations in cotton-top tamarins (Saguinus oedipus) are reported here. Monkeys were shown 1?+?1 events in which objects were placed behind a screen, 1 by 1. The screen was removed, revealing consistent (2 objects) and inconsistent (1, 3, or 1 large object twice the mass of original object) outcomes. In all studies, monkeys looked longer at the inconsistent than at the consistent outcome. When the monkeys view a 1?+?1 operation, they expect exactly 2 objects. It is likely that these numerical representations are spontaneously available to a variety of primate species and could provide a foundation on which humans' number sense was constructed over evolution and development. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Characterization of Clay Particle Surfaces for Contaminant Sorption in Soil Barriers Using Flow Microcalorimetry

Clays such as kaolinite and bentonite are widely used in various industries as sorbents. The sorptive characteristics of clays are exploited when they are used in contaminant barrier systems. To use clays effectively, their surface characteristics need to be known; especially, when they are used for contaminant sorption. Available surface area of clay minerals and the characteristics that depend on it are very sensitive to environmental changes such as those that can be induced by changes in the composition of pore fluid. Flow microcalorimetry with a down-stream concentration was used to determine the heats and amounts of adsorption of acids and bases on the clays. Test results presented herein revealed that both kaolinite and bentonite exhibit significantly different adsorption isotherms and heat of wetting under high pH and low pH conditions. Kaolinite has the capacity to adsorb both acidic and basic molecules almost equally. However, it has a tendency to adsorb more base than acid because of its stronger complexation capacity with acids than with bases. On the other hand, bentonite has a tendency to adsorb more acidic than basic molecules per gram. These results also indicate that both kaolinite and bentonite have different heats of wetting characteristics. As the concentrations of the acids and bases increase, the heat of wetting of kaolinite decreases while that of bentonite increases.