Poly(amido amine) (PAMAM) dendrimers have previously been shown, as cationic condensing agents of DNA, to have high potential for nonviral gene delivery. This study addresses two key issues for gene delivery: the interaction of the biomembrane with (i) the condensing agent (the cationic PAMAM dendrimer) and (ii) the corresponding dendrimer/DNA aggregate. Using in situ null ellipsometry and neutron reflection, parallel experiments were carried out involving dendrimers or generations 2 (G2), 4 (G4), and 6 (G6). The study demonstrates that free dendrimers of all three generations were able to traverse supported palmitoyloleoylphosphatidylcholine (POPC) bilayers deposited on silica surfaces. The model biomembranes were elevated front the solid surfaces upon dendrimer penetration, which offers a promising new way to generate more realistic model biomembranes where the contact with the supporting surface is reduced and where aqueous cavities are present beneath the bilayer. The largest dendrimer (GO) induced partial bilayer destruction directly upon penetration, whereas the smaller dendrimers (G2 and G4) leave the bilayer intact, so we propose that lower generation dendrimers have greater potential as transfection mediators. In addition to the experimental observations, coarse-grained simulations on the interaction between generation 3 (03) dendrimers and POPC bilayers were performed in the absence and presence of a bilayer-supporting negatively charged surface that emulates the support. The simulations demonstrate that G3 is transported across free-standing POPC bilayers by direct penetration and not by endocytosis. The penetrability was, however, reduced in the presence of a surface, indicating that the membrane transport observed experimentally was not driven solely by the surface. The experimental reflection techniques were also applied to dendrimer/DNA aggregates of charge ratio = 0.5, and while G2/DNA and G4/DNA aggregates interact with POPC bilayers. G6/DNA displays no such interaction. These results indicate that, in contrast to free dendrimer molecules, dendrimer/DNA aggregates of low charge ratios are not able to traverse a membrane by direct penetration.
The present paper investigates the structure and composition of grafted sodium hyaluronanat a solid-liquid interface using neutron reflection. The solvated polymer at the surface could be described with a density profile that decays exponentially towards the bulk solution. The density profileof the polymer varied depending on the deposition protocol. A single-stage deposition resulted in denser polymer layers, while layers created with a two-stage deposition process were more diffuse and had an overall lower density. Despite the diffuse density profile, two-stage deposition leads to a highersurface excess. Addition of calcium ions causes a strong collapse of the sodium hyaluronan chains, increasing the polymer density near the surface. This effect is more pronounced on the sample prepared by two-stage deposition due to the initial less dense profile. This study provides an understanding at a molecular level of how surface functionalization alters the structure and howsurface layers respond to changes in calcium ions in the solvent.
Polysaccharides are known to modify binding of proteins at interfaces and this paper describes studies of these interactions and how they are modified by pH. Specifically, the adsorption of human serum albumin on to polystyrene latex and to silica is described, focusing on how this is affected by hyaluronan. Experiments were designed to test how such binding might be modified under relevant physiological conditions. Changes in adsorption of albumin alone and the co-adsorption of albumin and hyaluronan are driven by electrostatic interactions. Multilayer binding is found to be regulated by the pH of the solution and the molecular mass and concentration of hyaluronan. Highest adsorption was observed at pH below 4.8 and for low molecular mass hyaluronan (<= 150 kDa) at concentrations above 2 mg ml(-1). On silica with grafted hyaluronan, albumin absorption is reversed by changes in solvent pH due to their strong electrostatic attraction. Albumin physisorbed on silica surfaces is also rinsed away with dilute hyaluronan solution at pH 4.8. The results demonstrate that the protein adsorption can be controlled both by changes of pH and by interaction with other biological macromolecules.
Knowledge of the structure of a biomaterial is usually vital to control its function. This article provides a structural characterization of a hyaluronan scaffold that has demonstrated good biocompatibility and is used to induce bone regeneration. Hyaluronan hydrogels are appealing materials that can function as a matrix to incorporate both organic and inorganic substances to enhance tissue growth. Because of the intrinsic properties of this swollen matrix, one needs a very sensitive technique that can be applied in situ to determine the organization of the polymers in a gel. Small-angle neutron scattering is used to determine the characteristics of the inhomogeneous structure of the hydrogel both with and without added particles. The results are interpreted using models of structure with two length scales that are beyond the traditional picture of homogeneous gels. The observed structure and the dimensions can explain the previously reported rheological properties of gels containing different amount of polymers. Hydroxyapatite nanoparticles added to the gel are frozen in the gel matrix. We are able to determine the distribution and shape of these particles as they aggregate around the polymer chains. We have also concluded, in this case, that the particle structure is concentration independent. Information about the nanostructure for an applicable biomaterial guides the formulation, preparation, and use that should lead to further understanding of its exploitation.
Hyaluronan based hydrogel coatings can mimic extracellular matrix components and incorporate growth factors that can be released during a progressive degradation while new tissue regenerates. This paper describes a structural characterization of a hydrogel coating made of modified hyaluronan polymers and how these coatings interact with bone morphogenetic protein-2 (BMP-2). Quartz crystal microbalance and neutron reflectivity measurements were used for in-situ, real-time measurements of the adsorption properties of polymers and proteins on smooth titanium oxide surfaces that mimic implant products in orthopedics. The adsorption of BMP-2 on a bare titanium oxide surface is compared to that on titanium oxide coated with different chemically modified hyaluronan, the most important being hyaluronan with bisphosphonate groups (HA-BP). The subsequent release of the BMP-2 from these hydrogel coatings could be triggered by calcium ions. The amount of adsorbed protein on the surfaces as well as the amount of released protein both depend on the type of hyaluronan coating. We conclude that HA-BP coated titanium oxide surfaces provide an excellent material for growth factor delivery in-vivo.
Films formed from saliva on surfaces are important for the maintenance of oral health and integrity by protection against chemical and/or biological agents. The aim of the present study was to investigate adsorbed amounts, thickness, and structure of films formed from human whole saliva on alumina surfaces by means of in situ ellipsometry, neutron reflectivity, and atomic force microscopy. Alumina (Al2O3, synthetic sapphire) is a relevant and interesting substrate for saliva adsorption studies as it has an isoelectric point close to that of tooth enamel. The results showed that saliva adsorbs rapidly on alumina. The film could be modeled in two layers: an inner and dense thin region that forms a uniform layer and an outer, more diffuse and thicker region that protrudes toward the bulk of the solution. The film morphology described a uniformly covering dense layer and a second outer layer containing polydisperse adsorbed macromolecules or aggregates.
A comprehensive description of the electrochemical processes in the positive electrodeof lithium–sulfur batteries is critical to the enhancement of sulfur utilization. However,the discharge mechanisms are complicated due to the various reactions in multiplephases and the tortuosity of the highly porous carbon matrix. While previous studieshave focused on the precipitation of Li2S, the effect of the limited mass transport insidethe micro-/mesopores of an electrode with optimized surface area have largely beenneglected. In this work, operando small-angle scattering with three different contrasts,and wide-angle scattering, has been performed with simultaneous resistance measure-ment of internal and diffusion. The results indicate that both electrode passivation andcomplete pore blockage are unlikely since the precipitates are surrounded by the elec-trolyte and grow mostly in number, not in size. The difference between the small- andwide-angle scattering reveals the amorphous discharge products at a low C-rate. Furtheranalyses demonstrate the correlation between the diffusion resistance and the contrastin the mesopores at the end of discharge, which suggests that Li-ion deficiency is thelimiting factor of sulfur utilization at a medium C-rate.
A comprehensive description of electrochemical processes in the positive electrode of lithium-sulfur batteries is crucial for the utilization of active material. However, the discharge mechanisms are complicated due to various reactions in multiple phases and the tortuosity of the highly porous carbon matrix. In this work, simultaneous measurements of small-angle and wide-angle scattering and cell resistance are performed on operating lithium-sulfur cells. Results indicate that precipitates grow mostly in number, not in size, and that the structure of the carbon matrix is not affected. The comparison of the small-angle and wide-angle scattering reveals the amorphous discharge products found at a low discharge rate. Further analysis demonstrates the correlation between the diffusion resistance and the compositional change of electrolyte in the mesopores at the end of discharge, which suggests that Li-ion deficiency is the limiting factor for sulfur utilization at a medium discharge rate.
A homologous series of n-alkyl trimethylammonium bromide surfactants, H(CH2)(n)N+(CH3)(3) Br-, from C(10)TAB to C(18)TAB have been studied systematically in the bulk over a wide range of temperatures. Common features in the structures are identified, with packing dominated by the co-ordination of the cationic head groups with bromide anions and interdigitation of the hydrocarbon chains. This arrangement provides an explanation for the thin adsorbed bilayers that have been observed at various hydrophilic surfaces from aqueous solutions in previous studies. The molecular volumes and arrangement are comparable with structures of a number of different self-assembled amphiphiles. For these surfactants with bromide counter-ions, formation of crystal hydrates was not observed. The alkyl chains are highly mobile and at high temperatures a plastic phase is found for all materials with a transition enthalpy that is similar to the melting enthalpy of many long alkyl chains. Other unexpected phase transitions depend more markedly on the hydrocarbon chain length and evidently depend on delicate balances of the various contributions to the free energy.
We have designed, built, and validated a (quasi)-simultaneous measurement platform called NUrF, which consists of neutron small-angle scattering, UV-visible, fluorescence, and densitometry techniques. In this contribution, we illustrate the concept and benefits of the NUrF setup combined with high-performance liquid chromatography pumps to automate the preparation and measurement of a mixture series of Brij35 nonionic surfactants with perfluorononanoic acid in the presence of a reporter fluorophore (pyrene).
The crystallization behavior during low-temperature annealing of samples of the Zr59.3Cu28.8Al10.4Nb1.5 (at%) bulk metallic glass produced by suction casting and the laser powder bed fusion (LPBF) process was studied with small-angle neutron scattering (SANS), X-ray diffraction, and scanning electron microscopy. The in-situ SANS measurements during isothermal annealing reveal that the phase separation in the LPBF processed material proceeds at a smaller characteristic length-scale than the cast material. Quantitative analysis of the SANS data shows that, while the crystallization process in both materials proceeds through rapid nucleation followed by diffusion-limited growth, the LPBF processed material crystallizes with a smaller cluster size and at a higher rate. The smaller cluster size is attributed to the elevated oxygen content in the LPBF processed material which reduces the nucleation barrier and thus the thermal stability.
Hypothesis: Endogenous Amorphous Magnesium-Calcium Phosphates (AMCPs) form in the human body and, besides their biomedical implications, the development of effective stabilization strategies is an open challenge. An interesting approach consists of stabilizing amorphous phosphates with macromolecules that have beneficial effects from a nutritional/medical point of view, for a potential application of the hybrid particles in nutraceutics or drug delivery.
Experimental: We investigated the effect of proteins extracted from Moringa oleifera seeds (MO) on the features of synthetic analogs of AMCPs and on their crystallization pathway. The stability of the amorphous phase was studied using infrared spectroscopy and X-ray diffraction. To unravel the effect of the protein on the nano-scale structure of the inorganic particles, we also studied how MO affects the features of the amorphous phase using thermal analysis, small angle X-ray scattering and confocal Raman microscopy.
Findings: We observed that MO markedly delays the transition from amorphous to crystalline phosphate in a concentration-dependent fashion. Interestingly, MO not only enhances the lifetime of the amorphous phase, but also influences the type and amount of crystalline material formed. The results are relevant from both a fundamental and an applied perspective, paving the way for the use of these hybrids in the field of nutraceutics and drug delivery.
Plasticisers are widely used to provide desirable mechanical properties of many polymeric materials. These small molecule additives are also known to leach from the finished products, and this not only may modify the physical properties but the distribution of these materials in the environment and in the human body can cause long-term health concerns and environmental challenges. Many of these plasticisers are esters of polyvalent acids and phthalic acid has previously been predominant but various alternatives are now being more widely explored. The eventual distribution of these compounds depends not just on solubility in aqueous media and on vapour pressure but also on their interaction with other materials, particularly lipids and amphiphiles. This review provides an overview of both the basic physical data (solubility, partition coefficients, surface tension, vapour pressure) that is available in the literature and summarises what has been learnt about the molecular interactions of various plasticisers with surfactants and lipids.
The adsorption of sodium bis 2-ethylhexyl sulfosuccinate, NaAOT, to a sapphire surface from aqueous solution has been studied by neutron reflection at concentrations above the critical micelle concentration (cmc). Complementary measurements of the bulk structure were made with small-angle neutron scattering and grazing incidence small-angle neutron scattering. At a concentration of about 1% wt (10 X cmc), lamellar phase NaAOT was observed both at the surface and in the bulk. The structure seen at the interface for a solution of 2% wt NaAOT is a 35 +/- 2 angstrom thick bilayer adsorbed to the sapphire surface at maximum packing density, followed by an aligned stack of fluctuating bilayers of thickness 51 +/- 2 angstrom and with an area per molecule of 40 +/- 2 angstrom(2). Each bilayer is separated by a water: at 25 degrees C, this layer is 148 +/- 2 angstrom. A simple model for the reflectivity from fluctuating layers is presented, and for 2.0% wt NaAOT the fluctuations were found to have an amplitude of 25 +/- 5 angstrom. The temperature sensitivity of the structure at the surface was investigated in the range 15-30 degrees C. The effect of temperature was pronounced, with the solvent layer becoming thinner and the volume occupied by the NaAOT molecules in a bilayer increasing with temperature. The amplitude of the fluctuations, however, is approximately temperature independent in this range. The adsorption of NaAOT at the sapphire surface resembles that previously found at hydrophilic and hydrophobic silica surfaces. The coexisting bulk lamellar phase has a spacing of layers similar to that observed at the surface. These observations are an indication that the major driving force for adsorption is self-assembly, independent of the chemical nature of the interface.
We report on the formation of large two-dimensional domains (about 20 cm2) of oriented and ordered structures of polystyrene particles dispersed in water at a solid/liquid interface. Gentle flow of the dispersed sample into the holder at a shear strain rate of about 0.1 s−1 caused particles at the air/latex meniscus to self-assemble in a regular structure on both solid silica or alumina surfaces. Scattering experiments show that the particle separation at the surface was the same as in the bulk and determined by repulsion arising from the charges on the particles. Close-packed planes formed parallel to the interface.
Proteins extracted from the seeds of Moringa trees are effective flocculents for particles dispersed in water and are attractive as a natural and sustainable product for use in water purification. Studies with a model system consisting of polystyrene latex particles have shown that the protein adsorbs to the surface and causes flocculation as unusually dense aggregates. Small-angle neutron scattering that exploits contrast matching of deuterated latex particles dispersed in D2O to highlight bound protein has shown that the adsorbed amount reaches about 3 mg m(-2). The particles form very compact flocs that are characterized by fractal dimensions that approach the theoretical maximum of 3. Ultra small-angle neutron scattering allows these flocs to be characterized for a range of particle and protein concentrations. Proteins from two species of Moringa trees were investigated. The protein from Moringa stenopetala seeds gave rise to slightly lower fractal dimensions compared to Moringa oleifera, but still much larger than values observed for conventional ionic or polymeric flocculents that are in the range 1.75-2.3. Compact flocs are desirable for efficient separation of impurities and dewatering of sludge as well as other applications. A trend of increasing fractal dimension with particle concentration was observed when M. stenopetala seed protein was used and this resembles the behaviour predicted in Brownian dynamics simulation of flocculation.
Quartz crystal microbalance with dissipation (QCM-D) monitoring is used to investigate the adsorption processes at liquid-solid interfaces and applied increasingly to characterize viscoelastic properties of complex liquids. Here, we contribute new insights into the latter field by using QCM-D to investigate the structure near the interface and the high-frequency viscoelastic properties of charge-stabilized polystyrene particles (radius 37 nm) dispersed in water. The study reveals changes with increasing ionic strength and particle concentration. Replacing water with a dispersion is usually expected to give rise to a decrease in frequency, f. Increases in both f and dissipation, D, were observed on exchanging pure water for particle dispersions at a low ionic strength. The QCM-D data are well-represented by a viscoelastic model, with viscosity increasing from 1.0 to 1.3 mPa s as the particle volume fraction changes from 0.005 to 0.07. This increase, higher than that predicted for noninteracting dispersions, can be explained by the charge repulsion between the particles giving rise to a higher effective volume fraction. It is concluded that the polystyrene particles did not adhere to the solid surface but rather were separated by a layer of pure dispersion medium. The QCM-D response was successfully represented using a viscoelastic Kelvin-Voigt model, from which it was concluded that the thickness of the dispersion medium layer was of the order of the particle-particle bulk separation, in the range of 50-250 nm, and observed to decrease with both particle concentration and addition of salt. Similar anomalous frequency and dissipation responses have been seen previously for systems containing weakly adherent colloidal particles and bacteria and understood in terms of coupled resonators. We demonstrate that surface attachment is not required for such phenomena to occur, but that a viscoelastic liquid separated from the oscillating surface by a thin Newtonian layer gives rise to similar responses.
The three-dimensional crystal structure of charge stabilised polystyrene latex in deionised water was investigated by small-angle neutron diffraction. Crystallisation with a grain size of approximately 1 x 1 cm(2) was observed when the sample was flowed gently in to a 2 mm path cell. Bragg scattering peaks arising from the structure were observed under rotation about three perpendicular axes of the sample. The diffraction patterns indicate clearly that there is a cubic close packed structure with a 110 axis perpendicular to the cell wall. Rotations in small steps show large changes and indicate that the crystal is well oriented and has three-dimensional order. The crystal orientation was controlled by the meniscus and direction of flow when filling the cell.
Aerosol-OT (sodium bis 2-ethylhexyl sulfosuccinate or NaAOT) adsorbs to hydrophilic sapphire solid surfaces The structure of the formed bilayer has been determined over the concentration range 0 2-7 4mM NaAOT It was found that the hydrocarbon tails pack at maximum packing limit at very low concentrations, and that the thickness of the bilayer was concentration-independent The adsorption was found to increase with concentration, with the surfactant molecules packing closer laterally The area per molecule was found to change from 138 +/- 25 to 51 +/- 4 angstrom(2) over the concentration range studied, with the thickness of the layer being constant at 33 2 A Addition of small amounts of salt was found to increase the surface excess, with the bilayer being thinner with a slightly larger area per molecule Addition of different salts of the same valency was found to have a very similiar effect, as had the addition of NaOH and HCl Hence, the effects of adding acid or base should be considered an effect of ionic strength rather than an effect of pH Adsorption of NaAOT to the sapphire surface that carries an opposite charge to the anionic surfactant is similar in many respects to the adsorption reported previously for hydrophilic and hydrophobic silica surfaces This suggests that the adsorption of NaAOT to a sui face is driven primarily by NaAOT self-assembly rather than effects of electrostatic at to the interface
The benefits of simultaneous studies of adsorbed layers and bulk structures are shown for solutions of the surfactant Aerosol-OT. Above the critical micelle concentration, Aerosol-OT forms an aligned lamellar phase at the sapphire/solution interface which is in equilibrium with a bulk phase that consists of coexisting micellar solution and dispersed lamellar phase. Measurements of the aligned surface layers and the bulk scattering from a 2% wt solution by grazing incidence and small-angle neutron scattering show that the bulk consist of lamellar structures with the same d-spacing as seen at the surface but without the surface induced alignment. The surface lamellar structure corresponds to a 10% volume fraction for a 2% wt bulk which implies that there must be coexistence of regions of different concentration. Scattering patterns measured in grazing incidence geometry clearly show the relative contributions from small-angle scattering and specular reflectivity.
Lung surfactant protein B (SP-B) is an essential protein found in the surfactant fluid at the air water interface of the lung. Exposure to the air pollutant ozone could potentially damage SP-B and lead to respiratory distress. We have studied two peptides, one consisting of the N-terminus of SP-B [SP-B(1-25)] and the other a construct of the N- and C-termini of SP-B [SP-B-(1-25,B-63-78)], called SMB. Exposure to dilute levels of ozone (similar to 2 ppm) of monolayers of each peptide at the air water interface leads to a rapid reaction, which is evident from an increase in the surface tension. Fluorescence experiments revealed that this increase in surface tension is accompanied by a loss of fluorescence from the tryptophan residue at the interface. Neutron and X-ray reflectivity experiments show that, in contrast to suggestions in the literature, the peptides are not solubilized upon oxidation but rather remain at the interface with little change in their hydration. Analysis of the product material reveals that no cleavage of the peptides occurs, but a more hydrophobic product is slowly formed together with an increased level of oligomerization. We attributed this to partial unfolding of the peptides. Experiments conducted in the presence of phospholipids reveal that the presence of the lipids does not prevent oxidation of the peptides. Our results strongly suggest that exposure to low levels of ozone gas will damage SP-B, leading to a change in its structure. The implication is that the oxidized protein will be impaired in its ability to interact at the air water interface with negatively charged phosphoglycerol lipids, thus compromising what is thought to be its main biological function.
Exposure to the secondary pollutant ozone in ambient air is associated with adverse health effects when inhaled. In this work we use surface pressure measurements, combined with X-ray and neutron reflection, to observe changes in a layer of lung surfactant at the air water interface when exposed to gas phase ozone. The results demonstrate that the layer reacts with ozone changing its physical characteristics. A slight loss of material, a significant thinning of the layer and increased hydration of the surfactant material is observed. The results support the hypothesis that unsaturated lipids present in lung surfactant are still susceptible to rapid reaction with ozone and the reaction changes the properties of the interfacial layer.
The structural changes that cause the change in interlayer spacing of a surfactant-templated zirconium oxide film have been studied using neutron diffractometry. We report that the film after drying on a glass substrate swells slightly through the addition of benzene by up to 4 Å on a lattice parameter of about 36 Å. The (0 0 1) and (0 0 2) diffraction peak widths, positions and areas of a swollen film were monitored as a function of benzene desorption. Disorder of the lamellar mesophase is considered as a cause of the observed effects on the diffraction signals.
Neutron reflectometry has been used to study the radical initiated oxidation of a monolayer of the lipid 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) at the air–solution interface by aqueous-phase hydroxyl, sulfate, and nitrate radicals. The oxidation of organic films at the surface of atmospheric aqueous aerosols can influence the optical properties of the aerosol and consequently can impact Earth’s radiative balance and contribute to modern climate change. The amount of material at the air–solution interface was found to decrease on exposure to aqueous-phase radicals which was consistent with a multistep degradation mechanism, i.e., the products of reaction of the DSPC film with aqueous radicals were also surface active. The multistep degradation mechanism suggests that lipid molecules in the thin film degrade to form progressively shorter chain surface active products and several reactive steps are required to remove the film from the air–solution interface. Bimolecular rate constants for oxidation via the aqueous phase OH radical cluster around 1010 dm3 mol–1 s–1. Calculations to determine the film lifetime indicate that it will take ∼4–5 days for the film to degrade to 50% of its initial amount in the atmosphere, and therefore attack by aqueous radicals on organic films could be atmospherically important relative to typical atmospheric aerosol lifetimes.
The heterogeneous oxidation of thin films of organic material extracted from real aerosol and sea-water samples was studied at the air-water interface using X-ray reflectivity. Oxidation of thin films of organic material extracted from real aerosol and sea-water is important in further understanding the impact of coated aerosols on the climate of the Earth. Surface active insoluble organic material extracted from the atmosphere was found to form stable films at the air-water interface (thickness measured as 10-14 nm). On exposure of the films to gas-phase ozone, no reaction (or change in the relative scattering length of the interface) was observed, indicating a potential lack of unsaturated organic material in the samples. Gas chromatography and electrospray ionization mass spectrometry showed the presence of saturated compounds in the samples. It is therefore proposed that the amount of unsaturated compounds as compared to saturated compounds in the atmospheric material is so low that the mass spectrometry analyses, as well as gas-phase oxidation are dominated by saturated material. A reaction was observed on exposure of the same films to aqueous phase hydroxyl and nitrate radicals and a film thinning mechanism is proposed to explain the change in scattering length of the film at the air-water interface. It can be suggested tentatively that oxidation by gas-phase ozone is not important in the atmosphere for organic films on aqueous atmospheric aerosol and that further studies should focus on radical induced oxidation of saturated organic material instead of unsaturated proxies that are typically studied.
The reactions between atmospheric oxidants and organic amphiphiles at the air water interface of an aerosol droplet may affect the size and critical supersaturation required for cloud droplet formation. We demonstrate that no reaction occurs between gaseous nitrogen dioxide (1000 ppm in air) and a monolayer of an insoluble amphiphile, oleic acid (cis-9-octadecenoic acid), at the air water interface which removes material from the air water interface. We present evidence that the NO2 isomerises the cis-9-octadecenoic (oleic) acid to trans-9-octadecenoic (elaidic) acid. The study presented here is important for future and previous studies of (1) the reaction between the nitrate radical, NO3, and thin organic films as NO2 is usually present in high concentrations in these experimental systems and (2) the effect of NO2 air pollution on the unsaturated fatty acids and lipids found at the air liquid surface of human lung lining fluid.
Organic films that form on atmospheric particulate matter change the optical and cloud condensation nucleation properties of the particulate matter and consequently have implications for modern climate and climate models. The organic films are subject to attack from gas-phase oxidants present in ambient air. Here we revisit in greater detail the oxidation of a monolayer of oleic acid by gas-phase ozone at the air-water interface as this provides a model system for the oxidation reactions that occur at the air-water interface of aqueous atmospheric aerosol. Experiments were performed on monolayers of oleic acid at the air-liquid interface at atmospherically relevant ozone concentrations to investigate if the viscosity of the sub-phase influences the rate of the reaction and to determine the effect of the presence of a second component within the monolayer, stearic acid, which is generally considered to be non-reactive towards ozone, on the reaction kinetics as determined by neutron reflectometry measurements. Atmospheric aerosol can be extremely viscous. The kinetics of the reaction were found to be independent of the viscosity of the sub-phase below the monolayer over a range of moderate viscosities, eta/eta water = 1.0-7.2, demonstrating no involvement of aqueous sub-phase oxidants in the rate determining step. The kinetics of oxidation of monolayers of pure oleic acid were found to depend on the surface coverage with different behaviour observed above and below a surface coverage of oleic acid of similar to 1 x 10(18) molecule m(-2). Atmospheric aerosol are typically complex mixtures, and the presence of an additional compound in the monolayer that is inert to direct ozone oxidation, stearic acid, did not significantly change the reaction kinetics. It is demonstrated that oleic acid monolayers at the air-water interface do not leave any detectable material at the air-water interface, contradicting the previous work published in this journal which the authors now believe to be erroneous. The combined results presented here indicate that the kinetics, and thus the atmospheric chemical lifetime for unsaturated surface active materials at the air-water interface to loss by reaction with gas-phase ozone, can be considered to be independent of other materials present at either the air-water interface or in the aqueous sub-phase.
The oxidation of organic films on cloud condensation nuclei has the potential to affect climate and precipitation events. In this work we present a study of the oxidation of a monolayer of deuterated oleic acid (cis-9-octadecenoic acid) at the air-water interface by ozone to determine if oxidation removes the organic film or replaces it with a product film. A range of different aqueous sub-phases were studied. The surface excess of deuterated material was followed by neutron reflection whilst the surface pressure was followed using a Wilhelmy plate. The neutron reflection data reveal that approximately half the organic material remains at the air-water interface following the oxidation of oleic acid by ozone, thus cleavage of the double bond by ozone creates one surface active species and one species that partitions to the bulk (or gas) phase. The most probable products, produced with a yield of similar to(87 +/- 14)%, are nonanoic acid, which remains at the interface, and azelaic acid (nonanedioic acid), which dissolves into the bulk solution. We also report a surface bimolecular rate constant for the reaction between ozone and oleic acid of (7.3 +/- 0.9) x 10(-11) cm(2) molecule s(-1). The rate constant and product yield are not affected by the solution sub-phase. An uptake coefficient of ozone on the oleic acid monolayer of similar to 4 x 10(-6) is estimated from our results. A simple Kohler analysis demonstrates that the oxidation of oleic acid by ozone on an atmospheric aerosol will lower the critical supersaturation needed for cloud droplet formation. We calculate an atmospheric chemical lifetime of oleic acid of 1.3 hours, significantly longer than laboratory studies on pure oleic acid particles suggest, but more consistent with field studies reporting oleic acid present in aged atmospheric aerosol.
An extract from the seeds of the Moringa oleifera tree that is principally a low molecular mass protein is known to be efficient as a coagulating agent for water treatment. The present paper investigates the adsorption of the purified protein to silica interfaces in order to elucidate the mechanism of its function as a flocculent. Neutron reflection permits the determination of the structure and composition of interfacial layers at the solid/solution interface. Dense layers of protein with about 5.5 mg m(-2) were found at concentrations above 0.025% wt. The overall thickness with a dense layer in excess of 60 angstrom at 0.05 wt % suggests strong co-operative binding rather than single isolated molecules. All ionic surfactant, sodium dodecyl sulfate, was also seen to coadsorb. This strong adsorption of protein in combination with the tendency for the protein to associate suggests a mechanism for destabilizing particulate dispersions to provide filterable water. This call occur even for the protein that has previously been identified as being of low mass (about 7 kDaltons) and thus is unlikely to be efficient in bridging or depletion flocculation.
The paper describes the adsorption of purified protein from seeds of Moringa olelfera to a sapphire interface and the effects of addition of the anionic surfactant sodium dodecylsulfate (SOS) and the cationic surfactant hexadecyltrimethylammonium bromide (CTAB). Neutron reflection was used to determine the structure and composition of interfacial layers adsorbed at the solid/solution interface. The maximum surface excess of protein was found to be about 5.3 mg m(-2). The protein does not desorb from the solid/liquid interface when rinsed with water. Addition of SDS increases the reflectivity indicating co-adsorption. It was observed that CTAB is able to remove the protein from the interface. The distinct differences to the behavior observed previously for the protein at the silica/water interface are identified. The adsorption of the protein to alumina in addition to other surfaces has shown why it is an effective flocculating agent for the range of impurities found in water supplies. The ability to tailor different surface layers in combination with various surfactants also offers the potential for adsorbed protein to be used in separation technologies.
Studies with a model system consisting of polystyrene latex particles showed that the protein from seeds of Moringa trees adsorbs to the surface and causes flocculation as unusually dense aggregates. In this study, electrolytes sodium chloride (NaCI), ferric chloride (FeCl3) and aluminium sulfate (Al-2(SO4)(3)) have been used to aggregate model polystyrene particles. The study augments previous work using neutron scattering on the flocculation of polystyrene latex with protein from seeds of Moringa trees that had indicated higher floc dimension, df, values as the concentration of particles increased. The measurements were made using ultra small-angle neutron scattering. Generally the fractal dimension, and thus the floc density, increased with particle concentration and salt concentration. Flocculation was apparent at much lower concentrations of FeCl3 and Al-2(SO4)(3) than of NaCI. The values of df were found not to simply scale with ionic strength for the three electrolytes studied with FeCl3 being the most effective flocculating agent.
Protein extracted from Moringa oleifera (MO) seeds has been advocated as a cheap and environmental friendly alternative to ionic flocculants for water purification. However, the nature and mechanism of its interaction with particles in water, as well as with dissolved surface-active molecules, are not well understood. In this article, we report studies of the protein and its interaction with four surfactants using dynamic light scattering (DLS), zeta-potential and turbidity measurements. Zeta-potential measurements identified points of charge reversal and the turbidity and DLS measurements were used to characterize the microstructure and size of protein-surfactant complexes. From the points of charge reversal, it was estimated that 7 anions are required to neutralize the positive charges of each protein molecule at pH 7. For protein mixtures with sodium dodecyl sulfate and dodecyl di-acid sodium salt, the peak in turbidity corresponds to concentrations with a large change in zeta-potential. No turbidity was observed for protein mixtures with either the nonionic surfactant Triton X-100 or the zwitterionic surfactant N-dodecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate. Changes of pH in the range 410 have little effect on the zeta-potential, turbidity, and the hydrodynamic radius reflecting the high isoelectric point of the protein. Addition of small amounts of salt has little effect on the size of protein in solution. These results are discussed in the context of the use of the MO protein in water treatment.
Commissioning results of a liquid sample cell for X-ray reflectivity studies with an in situ applied electrical field are presented. The cell consists of a Plexiglas container with lateral Kapton windows for air-liquid and liquid-liquid interface studies, and was constructed with grooves to accept plate electrodes on the walls parallel to the direction of the beam. Both copper and ITO plate electrodes have been used, the latter being useful for simultaneous optical studies. Commissioning tests were made at the I07 beamline of the Diamond Light Source.
The adsorption of the anionic surfactants, lithium, sodium and cesium dodecylsulfates, and sodium decylsulfonate, on the positively charged C-plane (0001) of sapphire (alumina) has been measured using neutron reflection. For each of the four surfactants there is a strong maximum in the adsorption at about the critical micelle concentration. The maximum becomes more marked from lithium to cesium. The measurements were reproduced over a range of different physical conditions and could not be accounted for in terms of impurities. The maximum is explained quantitatively by using the combination of a mass action model to calculate the mean activity of the surfactant, and a cooperative model of the adsorption (Frumkin), in which saturation of the layer is not attained until well above the critical micelle concentration.
The adsorption of the non-ionic surfactants tetraoxyethylene glycol monododecyl ether (C12EO4), pentaoxyethylene glycol monododecyl ether (C12EO5), and hexaoxyethylene glycol monododecyl ether (C12EO6) to single crystal sapphire substrates has been studied using specular neutron reflection for solutions at the critical micelle concentration. The effects of temperature and pH of the solutions were studied as well as the differences between two different crystal faces, the C and the R planes. At neutral pH, significant adsorption was only observed when the temperature was raised above the cloud temperature. This adsorption was reversible and surfactant was displaced on cooling. Reducing the pH to 3 results in significantly increased adsorption of C12EO5 at 25 degrees C with a central layer consisting mainly of surfactant (about 90%) on the C-plane substrate. A slightly smaller surface excess was observed for the R-plane. This contrasts with the significantly lower density observed even at high temperatures at neutral pH on both substrates. The results suggest that for neutral solutions surfactant association above the cloud point is the primary driving force for adsorption. At low pH, specific interactions with protonated surfaces are important. The structures of the highly hydrated layers are similar to those found for the surfactants at hydrophilic silica surfaces.
The role of ionic interactions between sodium dodecyl sulfate, SDS, and sapphire surfaces have been studied using specular neutron reflection to determine the structure and composition of adsorbed surfactant layers. Increasing the pH of the solution from 3 to 9 reduces the adsorption by reversing the charge of the alumina. This occurs at lower pH for the R-plane (1 (1) over bar 02) than the C-plane (0001), corresponding to the different points of zero charge. The largest surface excess is about 6.5 mu mol m(-2), the thickness of the adsorbed layer is about 24 angstrom and it contains roughly 20% water. The hydrocarbon tails of the surfactant molecules clearly interpenetrate rather than form an ordered bilayer. The structure is similar in either pure water or in 0.1 M NaCl when the surfactant is at the respective critical micelle concentration. Different structures were seen with lithium and cesium dodecyl sulfate. The CsDS forms dense layers with little or no hydration and a surface excess of about 10.5 mu mol m(-2). The metal cation strongly influences the hydration of the adsorbed surfactant. An overall picture of 'flattened micelles' for the structure of the adsorbed layer is observed.
In-situ time-resolved small-angle neutron scattering (SANS) has been applied for the study of the formation of mesoporous silica SBA-15. The advantage of neutron scattering, compared to X-ray scattering, is the possibility to contrast match i.e. highlight certain parts of the sample. Three different solvents with different scattering contrasts were used for each synthesis. Three different silica sources (tetramethyl orthosilicate, tetraethyl orthosilicate and tetrapropyl orthosilicate) were used and in three cases salts (sodium chloride or sodium bromide) were added prior to addition of the silica source. Hence, the effect of the silica sources and of the salts, on the formation of SBA-15 was investigated. The main focus was on the evolution of the ordered hexagonal structure i.e. investigation of the (10) Bragg peak. In synchrotron SAXS measurements the intensity of the (10) Bragg peak continuously increases during the measurement. However, in the SANS measurements the (10) Bragg peak area decreases with time. The decrease of the (10) peak is highly dependent on the solvent, a larger fraction of D2O in the solvent results in a bigger reduction. The decrease is also more pronounced when salt is present in the synthesis. The reduction of intensity reflects the chemistry in the wall and is explained by the compositional change in the wall during the maturation of the hexagonal order.
We report on the mechanism of growth of mesoporous silica (SBA-15, plane group p6m). In situ studies of the formation using ultrasmall angle X-ray scattering (USAXS) and small-angle X-ray scattering (SAXS) covering length scales from 5 to 10000 A, complemented with UV-vis and transmission electron microscopy (TEM), provide unique data on particle growth coupled with information regarding the progression of the mesostructure formation and the micellar evolution.
Comparison of melittin interaction with liposomes, bilayer disks and micelles showed that melittin binding to lipid aggregates is largely dictated by the amount of highly curved areas in the aggregates. The PEG-stabilised bilayer disks were characterised by a combination of small angle neutron scattering, cryo-transmission electron microscopy and dynamic light scattering. Importantly, the theoretically foreseen partial segregation of the lipid components, important for maintaining the structure of the bilayer disk, was confirmed. Steady state fluorescence spectroscopy indicated that melittin mainly resides at the rim of the bilayer disks. Results of the present study help increase the understanding of the mechanisms behind, and the physico-chemical factors affecting, melittin–lipid interaction. We suggest that bilayer disks, due to their stable structure, constitute interesting vehicles for transport of peptides that have high propensity to associate with lipid surfaces of high curvature.
Seed extracts from Moringa oleifera are of wide interest for use in water purification where they can play an important role in flocculation; they also have potential as anti-microbial agents. Previous work has focused on the crude protein extract. Here we describe the detailed biophysical characterization of individual proteins from these seeds. The results provide new insights relating to the active compounds involved. One fraction, designated Mo-CBP3, has been characterized at a molecular level using a range of biochemical and biophysical techniques including liquid chromatography, X-ray diffraction, mass spectrometry, and neutron reflection. The interfacial behavior is of particular interest in considering water purification applications and interactions with both charged (e.g. silica) and uncharged (alumina) surfaces were studied. The reflection studies show that, in marked contrast to the crude extract, only a single layer of the purified Mo-CBP3 binds to a silica interface and that there is no binding to an alumina interface. These observations are consistent with the crystallographic structure of Mo-CBP3-4, which is one of the main isoforms of the Mo-CBP3 fraction. The results are put in context of previous studies of the properties of the crude extract. This work shows possible routes to development of separation processes that would be based on the specific properties of individual proteins.
Deuteration of biomolecules has a major impact on both quality and scope of neutron scattering experiments. Cholesterol is a major component of mammalian cells, where it plays a critical role in membrane permeability, rigidity and dynamics, and contributes to specific membrane structures such as lipid rafts. Cholesterol is the main cargo in low and high-density lipoprotein complexes (i.e. LDL, HDL) and is directly implicated in several pathogenic conditions such as coronary artery disease which leads to 17 million deaths annually. Neutron scattering studies on membranes or lipid-protein complexes exploiting contrast variation have been limited by the lack of availability of fully deuterated biomolecules and especially perdeuterated cholesterol. The availability of perdeuterated cholesterol provides a unique way of probing the structural and dynamical properties of the lipoprotein complexes that underly many of these disease conditions. Here we describe a procedure for in vivo production of perdeuterated recombinant cholesterol in lipid-engineered Pichia pastoris using flask and fed-batch fermenter cultures in deuterated minimal medium. Perdeuteration of the purified cholesterol was verified by mass spectrometry and its use in a neutron scattering study was demonstrated by neutron reflectometry measurements using the FIGARO instrument at the ILL.
Changes of scattering are observed as the grazing angle of incidence of an incoming beam increases and probes different depths in samples. A model has been developed to describe the observed intensity in grazing incidence small angle neutron scattering (GISANS) experiments. This includes the significant effects of instrument resolution, the sample transmission, which depends on both absorption and scattering, as well as the sample structure. The calculations are tested with self-organised structures of two colloidal samples with different size particles that were measured on two different instruments. The model allows calculations for various instruments with defined resolution and can be used to design future improved experiments. The possibilities and limits of GISANS for different studies are discussed using the model calculations.
Self-assembly is a characteristic property of soft matter. Understanding the factors which assist or perturb this process is of great importance in many biological and industrial processes. Amphiphiles self-assemble and order into a variety of structures including well-ordered lamellar phases. The present work uses neutron reflectometry and neutron scattering to explore the effects of both interface roughness and temperature on the lamellar-phase structure of a non-ionic surfactant at a solid/liquid interface. The structure of concentrated solutions of tetraethyleneglycol dodecyl ether has been compared against a smooth surface and one with a roughness of the order of the lamellar spacing. This has been done in order to investigate the role perturbations have on the overall lamellar order, when these have length scales of the order of the interactions between neighboring lamellae. The results showed that the surfactant forms a well-ordered and aligned structure at a smooth surface, extending to a depth of several micrometers from the interface. Increasing the temperature of the sample and subsequent cooling promotes alignment and increases the number of oriented layers at the surface. The same sample forms a significantly less aligned structure against a rough surface that does not align to the same extent, even after heating. The perturbation of the structure caused by thermal fluctuations was found to be much less than that imposed by a small surface roughness.
Self-assembly is a characteristic property of soft matter and understanding the factors which assist or perturb this process is of a great importance in many biological and industrial processes. Amphiphiles self-assemble and order into a variety of structures including well-ordered lamellar phases. The present work uses neutron reflectometry to explore the effects of both interface roughness and temperature on the lamellar-phase structure of a non-ionic surfactant at a solid/liquid interface. The structure of concentrated solutions of tetraethyleneglycol dodecyl ether has been compared against a smooth surface and one with a roughness of the order of the lamellar spacing. The results showed that the surfactant forms a well-order and aligned structure at smooth surface that extends to a depth of micrometers from the interface. Increasing the temperature of the sample and subsequent cooling helped the alignment and increased the number of oriented layers at the surface. The same sample formed a significantly less aligned structure at a rough surface that did not align to same extent after heating. The perturbation of the structure caused by thermal fluctuations was found to be much less than that imposed by a small surface roughness.
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