Effect of benzocaine and propranolol on phospholipid-based bilayers.

Cell membranes play a fundamental role in protecting the cell from its surroundings, in addition to hosting many proteins with fundamental biological tasks. A study of drug/lipid interactions is a necessary and important step in fully clarifying the role and action mechanism of active ingredients, and shedding light on possible complications caused by drug overdosage. In this paper, the influence of benzocaine and propranolol drugs on the structure of l-α-phosphatidylcholine-based membranes has been investigated by means of neutron reflectivity, grazing incidence small angle neutron scattering, and small/ultra-small angle neutron scattering. Investigations allowed discovering a stiffening of the membranes and the formation of stalks, caused by the presence of benzocaine. On the other hand, disordered bilayers (lamellar powders) and highly curved structures were found in the presence of propranolol. The results obtained may be rationalized in terms of the molecular structures of drugs and may serve as a starting point for explaining the toxic behavior in long-term and overdosage scenarios.

Theta and high-beta networks for feedback processing: a simultaneous EEG-fMRI study in healthy male subjects.

The reward system is important in assessing outcomes to guide behavior. To achieve these purposes, its core components interact with several brain areas involved in cognitive and emotional processing. A key mechanism suggested to subserve these interactions is oscillatory activity, with a prominent role of theta and high-beta oscillations. The present study used single-trial coupling of simultaneously recorded electroencephalography and functional magnetic resonance imaging data to investigate networks associated with oscillatory responses to feedback during a two-choice gambling task in healthy male participants (n=19). Differential associations of theta and high-beta oscillations with non-overlapping brain networks were observed: Increase of high-beta power in response to positive feedback was associated with activations in a largely subcortical network encompassing core areas of the reward network. In contrast, theta-band power increase upon loss was associated with activations in a frontoparietal network that included the anterior cingulate cortex. Trait impulsivity correlated significantly with activations in areas of the theta-associated network. Our results suggest that positive and negative feedback is processed by separate brain networks associated with different cognitive functions. Communication within these networks is mediated by oscillations of different frequency, possibly reflecting different modes of dopaminergic signaling.

Multi-stage freezing of HEUR polymer networks with magnetite nanoparticles.

We observe a change in the segmental dynamics of hydrogels based on hydrophobically modified ethoxylated urethanes (HEUR) when hydrophobic magnetite nanoparticles (MNPs) are embedded in the hydrogels. The dynamics of the nanocomposite hydrogels is investigated using dielectric relaxation spectroscopy (DRS) and neutron spin echo (NSE) spectroscopy. The magnetic nanoparticles within the hydrophobic domains of the HEUR polymer network increase the size of these domains and their distance. The size increase leads to a dilution of the polymers close to the hydrophobic domain, allowing higher mobility of the smallest polymer blobs close to the "center". This is reflected in the decrease of the activation energy of the ß-process detected in the DRS data. The increase in distance leads to an increase of the size of the largest hydrophilic polymer blobs. Therefore, the segmental dynamics of the largest blobs is slowed down. At short time scales, i.e. 10(-9) s < τ < 10(-3) s, the suppression of the segmental dynamics is reflected in the α-relaxation processes detected in the DRS data and in the decrease of the relaxation rate Γ of the segmental motion in the NSE data with increasing concentration of magnetic nanoparticles. The stepwise (multi-stage) freezing of the small blobs is only visible for the pure hydrogel at low temperatures. On the other hand, the glass transition temperature (Tg) decreases upon increasing the MNP loading, indicating an acceleration of the segmental dynamics at long time scales (τ∼ 100 s). Therefore, it would be possible to tune the Tg of the hydrogels by varying the MNP concentration. The contribution of the static inhomogeneities to the total scattering function Sst(q) is extracted from the NSE data, revealing a more ordered gel structure than the one giving rise to the total scattering function S(q), with a relaxed correlation length ξNSE = (43 ± 5) Å which is larger than the fluctuating correlation length from a static investigation ξSANS = (17.2 ± 0.3) Å.

E. coli imprinted nano-structured silica micro-granules by spray drying: optimization of calcination temperature.

We have synthesized nano-structured silica-Escherichia coli composite micro-granules by spray drying of mixed suspension of silica and E. coli through evaporation-induced assembly. Synthesized micro-granules were subjected to calcination in order to form shape-matched macro-pores by removing the bacterial cells. The optimization of calcination temperature is crucial because calcination process leads to two contrasting effects, namely, (i) removal of E. coli from the granules and (ii) alteration of mesoscopic structure in the silica network. We have used small-angle neutron scattering and thermo-gravimetric analysis to determine the optimum temperature for calcination of these granules. It was found that calcination in the temperature range of 200°C to 400°C removes the cells without significant alteration of the nano-structured silica network. However, beyond 500°C, calcination results significant coalescence between the silica particles. Calcination at 600°C eventually collapses the meso-pore network of silica interstices.

Polymer enrichment decelerates surfactant membranes near interfaces.

Close to a planar surface, lamellar structures are imposed upon otherwise bulk bicontinuous microemulsions. Thermally induced membrane undulations are modified by the presence of the rigid interface. While it has been shown that a pure membrane's dynamics are accelerated close to the interface, we observed nearly unchanged relaxation rates for membranes spiked with large amphiphilic diblock copolymers. An increase of the polymer concentration by a factor of 2-3 for the first and second surfactant membrane layers was observed. We interpret the reduced relaxation times as the result of an interplay between the bending rigidity and the characteristic distance of the first surfactant membrane to the rigid interface, which causes the hydrodynamic and steric interface effects described in Seifert's theory. The influence of these effects on decorated membranes yields a reduction of the frequencies and an amplification of the amplitudes of long-wavelength undulations, which are in accordance to our experimental findings.

Acceleration of membrane dynamics adjacent to a wall.

The dynamics of an induced lamellar microemulsion adjacent to a planar hydrophilic surface (45 ns) were found to be three times faster compared to the bicontinuous bulk structure (133 ns). For these investigations the grazing incidence technique for neutron spin echo spectroscopy has been developed to resolve the depth dependent near surface dynamics. The observation is rationalized in terms of membrane hydrodynamics, where the flow fields reflected by the surface lead to a crossover from classical to confined fluctuations, and faster dynamics on large length scales (also known as "lubrication") are predicted.

Hydration and interactions in protein solutions containing concentrated electrolytes studied by small-angle scattering.

During protein crystallization and purification, proteins are commonly found in concentrated salt solutions. The exact interplay of the hydration shell, the salt ions, and protein-protein interactions under these conditions is far from being understood on a fundamental level, despite the obvious practical relevance. We have studied a model globular protein (bovine serum albumin, BSA) in concentrated salt solutions by small-angle neutron scattering (SANS). The data are also compared to previous studies using SAXS. The SANS results for dilute protein solutions give an averaged volume of BSA of 91,700 Å(3), which is about 37% smaller than that determined by SAXS. The difference in volume corresponds to the contribution of a hydration shell with a hydration level of 0.30 g g(-1) protein. The forward intensity I(0) determined from Guinier analysis is used to determine the second virial coefficient, A(2), which describes the overall protein interactions in solution. It is found that A(2) follows the reverse order of the Hofmeister series, i.e. (NH(4))(2)SO(4) < Na(2)SO(4) < NaOAc < NaCl < NaNO(3) < NaSCN. The dimensionless second virial coefficient B(2), corrected for the particle volume and molecular weight, has been calculated using different approaches, and shows that B(2) with corrections for hydration and the non-spherical shape of the protein describes the interactions better than those determined from the bare protein. SANS data are further analyzed in the full q-range using liquid theoretical approaches, which gives results consistent with the A(2) analysis and the experimental structure factor.

Origin of buckling phenomenon during drying of micrometer-sized colloidal droplets.

The origin of the buckling of micrometer-sized colloidal droplets during evaporation-induced self-assembly (EISA) has been elucidated using electron microscopy and small-angle neutron scattering. Doughnut-like assembled grains with varying aspect ratios are formed during EISA at different physicochemical conditions. It has been revealed that this phenomenon is better explained by an existing hypothesis based on the formation of a viscoelastic shell of nanoparticles during drying than by other existing hypotheses based on the inertial instability of the initial droplets and hydrodynamic instability due to thermocapillary forces. This conclusion was further supported by the arrest of buckling through modification of the colloidal interaction in the initial dispersion.

Near-surface structure of a bicontinuous microemulsion with a transition region.

The lamellar ordering of bicontinuous microemulsions adjacent to a planar hydrophilic wall is investigated experimentally by grazing-incidence small-angle neutron scattering and theoretically by computer simulations. It is shown that precise depth information in neutron scattering can be obtained by tuning the scattering length density of the overall microemulsion. Neutron reflectometry completes the characterization. The nucleation of a lamellar phase at the wall is observed, and a perforated lamellar transition region is identified at the lamellar-microemulsion interface. The thickness of the lamellar region is about 400 Å, which corresponds to two bilayers.

Thermoreversible lysozyme hydrogels: properties and an insight into the gelation pathway.

The gelation behaviour of aqueous solutions of hen egg white lysozyme (HEWL) in the presence of 20 mM DTT in the concentration range 0.7 to 4.0 mM has been investigated using microDSC, FTIR, cryoTEM, SANS and oscillatory rheology. The macroscopic critical gelation concentration, Cgel, was found to be ∼ 3.0 mM. The disruption of the disulfide bonds by the DTT and the destabilisation of the protein were found to be a prerequisite for the formation of ß-sheet rich fibrils under the mild conditions used in this work. Using our methodology the hydrogels obtained have a pH of 7, hence are suitable for cell culture, and are also thermoreversible. The hydrogel melting temperature was found to increase with increasing concentration and a similar structure was observed across the concentration range investigated. Our results suggest these systems are composed of a well defined regular network where single ß-sheet rich fibrils (∼ 3 nm diameter) form initially, then two of these fibrils associate two-by-two to form junctions (∼ 6 nm diameter) and then on cooling further aggregate to form larger bundles of fibres. The network mesh size was found to decrease with increasing concentration. Our results suggest that below Cgel small unconnected gel-like aggregates exist that have a similar structure to the hydrogels obtained above Cgel. Using our data we propose a model for the denaturation and gelation behaviour of our system. During the first heating an α-helix to ß-sheet molecular transition for the protein conformation occurs resulting in ß-sheet rich fibrils forming through the self-assembly of ß-sheet rich denaturated proteins. At high temperature the solution contains ß-sheet rich fibrils with dissolved protein. On cooling an increase in the amount of ß-sheet was observed via FTIR suggesting that as the temperature is decreased more and more protein forms ß-sheet rich fibrils. At the gelation temperature these fibrils associate two-by-two to form the network junctions resulting in the macroscopic gelation of the sample. Our results suggest the network junctions are formed via specific hydrophobic interactions. The hydrogels elastic modulus was found to scale as C2.45 for C > Cgel.

SANS study of polymer-linked droplets.

We report experimental results obtained from SANS on microemulsion droplets connected by a telechelic polymer. Thanks to its ability to anchor droplets through its short stickers, the addition of this polymer leads to the formation of transient aggregates. Measurements were performed on samples at low surfactant content in such a way that the droplets appear to be isolated with a separation distance longer than the end-to-end distance of the polymer. The locally spherical structure of the micelles is unchanged in size upon polymer addition whereas the large rise in scattered intensity at low Q is due to the induced effective attractive interaction between droplets. The fitting model that we propose allows a quantitative description of the bridging effect.

Hydrodynamic effects in bicontinuous microemulsions measured by inelastic neutron scattering.

The dynamical properties of bicontinuous microemulsions have been studied with neutron spin echo spectroscopy around length scales corresponding to the correlation peak q(0). Comparison of samples with different contrasts for neutrons shed light on the two modes dominated either by variation of the oil/water difference or surfactant concentration in the hydrodynamic regime. The results have been compared to theoretical predictions of the relaxation rates of bicontinuous microemulsions by Nonomura and Ohta [M. Nonomura, T. Ohta, J. Chem. Phys. 110, 7516 (1999)]. The influence of modification of the surfactant layer bending constants in the microemulsion by addition of homopolymers (polyethylenepropylene: PEP(X) and polyethyleneoxide: PEO(X), X=5 kg/mol), dissolved in the oil phase and water, has been investigated.

Adsorption of supercritical CO2 in aerogels as studied by small-angle neutron scattering and neutron transmission techniques.

Small-angle neutron scattering (SANS) has been used to study the adsorption behavior of supercritical carbon dioxide (CO2) in porous Vycor glass and silica aerogels. Measurements were performed along two isotherms (T=35 and 80 degrees C) as a function of pressure (P) ranging from atmospheric up to 25 MPa, which corresponds to the bulk fluid densities ranging from rho(CO2) approximately 0 to 0.9 gcm3. The intensity of scattering from CO2-saturated Vycor porous glass can be described by a two-phase model which suggests that CO2 does not adsorb on the pore walls and fills the pore space uniformly. In CO2-saturated aerogels an adsorbed phase is formed with a density substantially higher that of the bulk fluid, and neutron transmission data were used to monitor the excess adsorption at different pressures. The results indicate that adsorption of CO2 is significantly stronger in aerogels than in activated carbons, zeolites, and xerogels due to the extremely high porosity and optimum pore size of these materials. SANS data revealed the existence of a compressed adsorbed phase with the average density approximately 1.07 gcm3, close to the density corresponding to closely packed van der Waals volume of CO2. A three-phase model [W. L. Wu, Polymer 23, 1907 (1982)] was used to estimate the volume fraction phi3 of the adsorbed phase as a function of the fluid density, and gave phi3 approximately 0.78 in the maximum adsorption regime around rho(CO2) approximately 0.374 gcm3. The results presented in this work demonstrate the utility of SANS combined with the transmission measurements to study the adsorption of supercritical fluids in porous materials.

Microemulsion efficiency boosting and the complementary effect. 1. Structural properties.

Amphiphilic diblock copolymers added to microemulsions proved to enhance the efficiency of surfactants dramatically. The complementary effect of homopolymers is considered in the current work. A possible application of the homopolymer addition could be the viscosity tuning of the microemulsion without changing the considered bicontinuous phase. Furthermore, (homo)polymers are added for many other reasons in technical applications. A theory by Eisenriegler predicts a decreased efficiency when homopolymers are added. In further experiments, the simultaneous addition of homopolymers and diblock copolymers probes whether the two opposite effects superpose and allow for a compensation. Then, efficiency and viscosity are adjustable independently. Experimentally, phase diagrams are investigated and the microscopic structure is measured by small-angle neutron scattering. Within the presented models, both experimental methods are compared and discussed on the basis of the surfactant membrane bending moduli. The homopolymer effect is about 7 times larger than that theoretically predicted, and the superposition of the two polymer effects allows for a compensation with an optionally tunable viscosity.

Density fluctuations near the liquid-gas critical point of a confined fluid.

We report the results of an experimental study of the effect of a dilute silica network on liquid-gas critical phenomena in carbon dioxide (CO2). Using small-angle neutron scattering, we measured the correlation length of the density fluctuations in bulk (xi(bulk)) and confined CO2 (xi(conf)) as a function of temperature and average fluid density. We find that quenched disorder induced by an aerogel suppresses density fluctuations: xi(conf) loses the Ising model divergence characteristic of xi(bulk) and does not exceed the size of pores in the homogeneous region.

Distribution and unspecific protein binding of the xenoestrogens bisphenol A and daidzein.

Physiological toxicokinetic (PT) models are used to simulate tissue burdens by chemicals in animals and humans. A prerequisite for a PT model is the knowledge of the chemical's distribution among tissues. This depends on the blood flow and also on the free fraction of the substance and its tissue:blood partition coefficients. In the present study we determined partition coefficients in human tissues at 37 degrees C for the two selected xenoestrogens bisphenol A (BA) and daidzein (DA), and their unspecific binding to human serum proteins. Partition coefficients were obtained by incubating blood containing BA or DA with each of the following tissues: brain, liver, kidney, muscle, fat, placenta, mammary gland, and adrenal gland. Blood samples were analysed by HPLC. For BA and DA, all partition coefficients in non-adipose tissues were similar (average values: BA 1.4, DA 1.2). However, the lipophilic properties of both compounds diverge distinctly. Fat:blood partition coefficients were 3.3 (BA) and 0.3 (DA). These values indicate that with the exception of fat both compounds are distributed almost equally among tissues. In dialysis experiments, the unspecific binding of BA and DA with human serum proteins was measured by HPLC. For BA, the total concentration of binding sites and the apparent dissociation constant were calculated as 2000 and 100 nmol/ml, respectively. Because of the limited solubility of DA, only the ratio of the bound to the free DA concentration could be determined and was found to be 7.2. These values indicate that at low concentrations only small percentages of about 5% (BA) and 12% (DA) are as unbound free fractions in plasma. Since only the unbound fraction can bind to the estrogen receptor, binding to serum proteins represents a mechanism that limits the biological response in target tissues.

A low-molecular-weight cytosolic inhibitor of the specific testosterone binding in bovine seminal vesicles.

Bovine seminal vesicle cytosol contains a low-molecular-weight and thermostable substance which specifically inhibits the binding of testosterone to its cognate receptor. The mass and the ether phospholipid structure of the inhibitor were elucidated by mass spectrometry. Saturation and binding experiments indicate that the inhibitor acts in a dose-dependent and competitive manner, altering the apparent dissociation constant (K(D)) while maintaining the number of androgen-binding sites (B(max)). Its possible role in the regulation of androgen binding activity is discussed.