Thermal Density Fluctuations and Polymorphic Phase Transitions of Ethane (C2D6) in the Gas/Liquid and Supercritical States.

The phase behavior of the liquid C2D6 below and above the critical point was investigated using small-angle neutron scattering (SANS) in temperature and pressure ranges from 10 to 45 °C and 20 to 126 bar, respectively. The scattering of thermal fluctuations of the molecular density was determined and thus the gas-liquid and Widom lines. At the same time, we observed additional scattering of droplets of more densely packed C2D6 molecules above the gas-liquid line and in the supercritical fluid regime from just below the critical point for all temperatures at about ΔP = 10 bar above the Widom line. This line is interpreted as the Frenkel line. These results are consistent with our previous studies on CO2 and thus indicate a universal phase behavior for monomolecular liquids below and above the critical point. The interpretation of the Frenkel line as the lower limit of a polymorphic phase transition is in contrast to the usual interpretation as the limit of a dynamic process. The correlation lengths (ξ) of the thermal density fluctuations at the critical point and at the Widom line are determined between 20 and 35 Å and thus in the range of the droplet radius between 60 and 80 Å. These long-range fluctuations appear to suppress the formation of droplets, which can only form at about 10 bar above the critical point and the Widom line when ξ becomes smaller than 10 Å.

Silica Fouling in Reverse Osmosis Systems-Operando Small-Angle Neutron Scattering Studies.

We present operando small-angle neutron scattering (SANS) experiments on silica fouling at two reverse osmose (RO) membranes under almost realistic conditions of practiced RO desalination technique. To its realization, two cells were designed for pressure fields and tangential feed cross-flows up to 50 bar and 36 L/h, one cell equipped with the membrane and the other one as an empty cell to measure the feed solution in parallel far from the membrane. We studied several aqueous silica dispersions combining the parameters of colloidal radius, volume fraction, and ionic strength. A relevant result is the observation of Bragg diffraction as part of the SANS scattering pattern, representing a crystalline cake layer of simple cubic lattice structure. Other relevant parameters are silica colloidal size and volume fraction far from and above the membrane, as well as the lattice parameter of the silica cake layer, its volume fraction, thickness, and porosity in comparison with the corresponding permeate flux. The experiments show that the formation of cake layer depends to a large extent on colloidal size, ionic strength and cross-flow. Cake layer formation proved to be a reversible process, which could be dissolved at larger cross-flow. Only in one case we observed an irreversible cake layer formation showing the characteristics of an unstable phase transition. We likewise observed enhanced silica concentration and/or cake formation above the membrane, giving indication of a first order liquid-solid phase transformation.

Polymorphic phase transition in liquid and supercritical carbon dioxide.

We present experiments on molecular density fluctuations in liquid and supercritical (SC) CO2 using small-angle neutron scattering. Thermal density fluctuations in SC-CO2 determine susceptibility and correlation length identifying the Widom line at their maxima. Droplet formation occurs at the gas-liquid line and between 20 and 60 bar above the Widom line, the corresponding borderline identified as the Frenkel line. The droplets start to form spheres of constant radius of ≈ 45 Å and transform into rods and globules at higher pressure. Droplet formation represents a liquid-liquid (polymorphic) phase transition of the same composition but different density, whose difference defines its order parameter. Polymorphism in CO2 is a new observation stimulating interesting discussions on the topics of gas-like to liquid-like transition in SC fluids and polymorphism since CO2 represents a "simple" van der Waals liquid in contrast to water, which is the most widely studied liquid showing polymorphism in its supercooled state.

Morphology of Thin Film Composite Membranes Explored by Small-Angle Neutron Scattering and Positron-Annihilation Lifetime Spectroscopy.

The morphology of thin film composite (TFC) membranes used in reverse osmosis (RO) and nanofiltration (NF) water treatment was explored with small-angle neutron scattering (SANS) and positron-annihilation lifetime spectroscopy (PALS). The combination of both methods allowed the characterization of the bulk porous structure from a few Å to µm in radius. PALS shows pores of 4.5 Å average radius in a surface layer of about 4 m thickness, which become 40% smaller at the free surface of the membranes. This observation may correlate with the glass state of the involved polymer. Pores of similar size appear in SANS as closely packed pores of 6 Å radius distributed with an average distance of 30 Å. The main effort of SANS was the characterization of the morphology of the porous polysulfone support layer as well as the fibers of the nonwoven fabric layer. Contrast variation using the media H2O/D2O and supercritical CO2 and CD4 identified the polymers of the support layers as well as internal heterogeneities.

Surface-Induced Silica Scaling during Brackish Water Desalination: The Role of Surface Charge and Specific Chemical Groups.

Silica scaling of membranes used in reverse osmosis desalination processes is a severe problem, especially during the desalination of brackish groundwater due to high silica concentrations. This problem limits the water supply in inland arid and semiarid regions. Here, we investigated the influence of surface-exposed organic functional groups on silica precipitation and scaling. A test solution simulating the mineral content of brackish groundwater desalination brine at 75% recovery was used. The mass and chemical composition of the precipitated silica was monitored using a quartz crystal microbalance, X-ray photoelectron spectroscopy, and infrared spectroscopy, showing that surfaces with positively charged groups induced rapid silica precipitation, and the rate of silica precipitation followed the order -NH2 ∼ -N+(CH3)3 > -NH2/-COOH > -H2PO3 ∼ -OH > -COOH > -CH3. Force vs distance AFM measurements showed that the adhesion energy between a silica colloid glued to AFM cantilever and the studied surfaces increased as the surface charge changed from negative to positive. Thus, for the first time direct measurements of molecular forces and specific chemical groups that govern silica scaling during brackish water desalination is reported here. The influence of the different functional groups and the effect of the surface charge on silica precipitation that were found here can be used to design membranes that resist silica scaling in membrane-based desalination processes.

Necklace-like microstructure in shallow-quenched aqueous solutions of poly(n-isopropylacrylamide), detected by advanced small-angle neutron scattering methods.

The microstructure of aqueous poly(N-isopropyl acrylamide) (PNIPA) gel and solution was investigated by small-angle neutron scattering (SANS) in the vicinity of the gel volume phase transition at TV (= 34 °C). The SANS technique was reinforced by refractive neutron lenses and perfect single crystals in order to get access to µm length scales. At 31 °C SANS shows Ornstein-Zernike (OZ) type scattering in the swollen gel which at 32 °C starts to deviate from the OZ-formalism, exhibiting excess scattering and at the wave number qc≅ 5 × 10-3Å-1 a crossover to Porod's asymptotic q-4 power law. For shallow quenches of ΔT < 1.0 K above TV the excess scattering intensity is further increasing whereas qc is shifting toward lower values. Based on this observation and analysis of the SANS q-profiles, we propose a necklace-like microstructure consisting of PNIPA-rich globules of R≅ 100 Å size which are connected by swollen PNIPA chains and stabilized for more than a day by pinning of chain connectivity. The formation of PNIPA globules near TV is discussed in terms of partially cooperative dehydration which is crucial to explain the "miscibility square phase behavior" of aqueous PNIPA solutions. Globule-like structure was also found in aqueous PNIPA solution of size slightly larger than in gels. At deeper quenches of gels above TV (ΔT > 1.0 K) the globules are aggregating to larger objects of R≅ 0.24 µm size as determined from a strong intensity upturn in the small q-region of USANS.

Ionic Dependence of Gelatin Hydrogel Architecture Explored Using Small and Very Small Angle Neutron Scattering Technique.

The hierarchical structure of gelatin hydrogels mimics a natural extracellular matrix and provides an optimized microenvironment for the growth of 3D structured tissue analogs. In the presence of metal ions, gelatin hydrogels exhibit various mechanical properties that are correlated with the molecular interactions and the hierarchical structure. The structure and structural response of gelatin hydrogels to variation of gelatin concentration, pH, or addition of metal ions are explored by small and very small angle neutron scattering over broad length scales. The measurements of the hydrogels reveal the existence of a two-level structure of colloid-like large clusters and a 3D cage-like gel network. In the presence of Fe3+ ions the hydrogels show a highly dense and stiff network, while Ca2+ ions have an opposite effect. The results provide important structural insight for improvement of the design of gelatin based hydrogels and are therefore suitable for various applications.

Densification of Supercritical Carbon Dioxide Accompanied by Droplet Formation When Passing the Widom Line.

Thermal density fluctuations of supercritical CO_{2} were explored using small-angle neutron scattering (SANS), whose amplitude (susceptibility) and correlation length show the expected maximum at the Widom line. At low pressure, the susceptibility is in excellent agreement with the evaluated values on the basis of mass density measurements. At about 20 bar beyond the Widom line, SANS shows the formation of droplets accompanied by an enhanced number density of the supercritical fluid. The corresponding borderline is interpreted as a Frenkel line separating gas- and liquidlike regimes.

Calcium phosphate scaling during wastewater desalination on oligoamide surfaces mimicking reverse osmosis and nanofiltration membranes.

Desalinated domestic wastewater is an indispensable water resource in arid regions; however, its recovery can be limited by calcium phosphate scaling and fouling of the membrane. Here we investigated calcium phosphate mineralization on oligoamide surfaces that mimics reverse osmosis (RO) and nanofiltration (NF) membrane surfaces. We used a solution that simulates desalination of secondary treated domestic wastewater effluents for calcium phosphate mineralization experiments with oligoamide-coated gold surfaces. Attenuated total reflection-Fourier transform infrared spectroscopy and energy dispersive spectrometry showed that calcium phosphate and carbonate precipitated on RO mimetic surfaces. The rate of precipitation on oligoamide sensors was monitored by a quartz crystal microbalance, showing that scaling was more intense on the RO than the NF mimetic surface and that excessive carboxyl functional groups on both surfaces promoted scaling. Filtration experiments of similar solutions with commercial membranes showed that scaling was more intense on the RO membranes than on the NF membranes, which supported the results obtained with the oligoamide model surfaces. The results of this study can be implemented in developing RO and NF membranes to prevent calcium phosphate scaling and consequently lower water-treatment costs of domestic wastewater treatment.

Biopolymer-induced calcium phosphate scaling in membrane-based water treatment systems: Langmuir model films studies.

Biofouling and scaling on reverse osmosis (RO) or nanofiltration (NF) membranes during desalination of secondary and tertiary effluents pose an obstacle that limits the reuse of wastewater. In this study we explored the mineral scaling induced by biopolymers originated from bacterial biofilms: bovine serum albumin (BSA), fibrinogen, lysozyme and alginic acid, as well as an extracts of extracellular polymeric substances (EPS) from bio-fouled RO membranes from wastewater treatment facility. Mineralization studies were performed on Langmuir films of the biopolymers deposited at the interface of a solution simulating RO desalination of secondary-treated wastewater effluents. All studied biopolymers and EPS induced heterogeneous mineralization of mainly calcium phosphate. Using IR spectroscopy coupled with systematic quantitative analysis of the surface pressure versus molecular-area isotherms, we determined the mineralization tendencies of the biopolymers to be in the order of: fibrinogen>lysozyme>BSA>alginic acid. The biopolymers and EPS studied here were found to be accelerators of calcium-phosphate mineralization. This study demonstrates the utilization of Langmuir surface-pressure area isotherms and a model solution in quantitatively assessing the mineralization tendencies of various molecular components of EPS in context of membrane-based water treatment systems.

Multifunctional layered magnetic composites.

A fabrication method of a multifunctional hybrid material is achieved by using the insoluble organic nacre matrix of the Haliotis laevigata shell infiltrated with gelatin as a confined reaction environment. Inside this organic scaffold magnetite nanoparticles (MNPs) are synthesized. The amount of MNPs can be controlled through the synthesis protocol therefore mineral loadings starting from 15 wt % up to 65 wt % can be realized. The demineralized organic nacre matrix is characterized by small-angle and very-small-angle neutron scattering (SANS and VSANS) showing an unchanged organic matrix structure after demineralization compared to the original mineralized nacre reference. Light microscopy and confocal laser scanning microscopy studies of stained samples show the presence of insoluble proteins at the chitin surface but not between the chitin layers. Successful and homogeneous gelatin infiltration in between the chitin layers can be shown. The hybrid material is characterized by TEM and shows a layered structure filled with MNPs with a size of around 10 nm. Magnetic analysis of the material demonstrates superparamagnetic behavior as characteristic for the particle size. Simulation studies show the potential of collagen and chitin to act as nucleators, where there is a slight preference of chitin over collagen as a nucleator for magnetite. Colloidal-probe AFM measurements demonstrate that introduction of a ferrogel into the chitin matrix leads to a certain increase in the stiffness of the composite material.

Synthesis and Characterization of Gelatin-Based Magnetic Hydrogels.

A simple preparation of thermoreversible gelatin-based ferrogels in water provides a constant structure defined by the crosslinking degree for gelatin contents between 6 and 18 wt%. The possibility of varying magnetite nanoparticle concentration between 20 and 70 wt% is also reported. Simulation studies hint at the suitability of collagen to bind iron and hydroxide ions, suggesting that collagen acts as a nucleation seed to iron hydroxide aggregation, and thus the intergrowth of collagen and magnetite nanoparticles already at the precursor stage. The detailed structure of the individual ferrogel components is characterized by small-angle neutron scattering (SANS) using contrast matching. The magnetite structure characterization is supplemented by small-angle X-ray scattering and microscopy only visualizing magnetite. SANS shows an unchanged gelatin structure of average mesh size larger than the nanoparticles with respect to gel concentration while the magnetite nanoparticles size of around 10 nm seems to be limited by the gel mesh size. Swelling measurements underline that magnetite acts as additional crosslinker and therefore varying the magnetic and mechanical properties of the ferrogels. Overall, the simple and variable synthesis protocol, the cheap and easy accessibility of the components as well as the biocompatibility of the gelatin-based materials suggest them for a number of applications including actuators.

Effects of biological molecules on calcium mineral formation associated with wastewater desalination as assessed using small-angle neutron scattering.

Calcium phosphate scale formation on reverse osmosis (RO) membranes is one of the main limitations on cost-effective desalination of domestic wastewater worldwide. It has been shown that organic agents affect mineralization. In this study, we explored mineralization in the presence of two biofilm-relevant organic compounds, the proteins bovine serum albumin (BSA) and lysozyme, in a simulated secondary effluent (SSE) solution using small-angle neutron scattering (SANS), and applied the results to analyses of mineral precipitation in RO desalination of secondary effluents of wastewater. The two proteins are prominent members of bacterial extracellular polymeric substances (EPSs), forming biofilms that are frequently associated with RO-membrane fouling during wastewater desalination. Laboratory experiments showed that both proteins in SSE solution are involved in complex mineralization processes. Only small portions of both protein fractions are involved in mineralization processes, whereas most of the protein fractions remain as monomers in solution. Contrast variation showed that composite particles of mineral and protein are formed instantaneously to a radius of gyration of about 300 Å, coexisting with particles of about µm size. After about one day, these large particles start to grow again at the expense of the 300 Å particles. The volume fraction of the 300 Å particles is of the order of 2 × 10(-4), which is too large to represent calcium phosphate such as hydroxyapatite as the only mineral present. Considering the data of mineral volume fraction obtained here as well as the solubility product of possible mineral polymorphs in the SSE solution, we suggest the formation of protein-mineral particles of hydroxyapatite and calcium carbonate during scale formation.

Fetuin-A is a mineral carrier protein: small angle neutron scattering provides new insight on Fetuin-A controlled calcification inhibition.

Clinical studies and animal experiments have shown that the serum protein fetuin-A is a highly effective inhibitor of soft tissue calcification. This inhibition mechanism was elucidated on the basis of an in vitro fetuin-A-mineral model system. In a previous study, we found that in a two-stage process ∼100-nm sized calciprotein particles (CPPs) were formed whose final stage was stabilized by a compact outer fetuin-A monolayer against further growth. Quantitative small-angle neutron scattering data analysis revealed that even at a fetuin-A concentration close to the stability limit, only approximately one-half of the mineral ions and only 5% of the fetuin-A were contained in the CPPs. To uncover the interplay of the remaining supersaturated mineral ion fraction and of the 95% non-CPP fetuin-A, we explored the fetuin-A monomer fraction in solution by contrast variation small-angle neutron scattering. Our results suggest that the mineral ions coalesce to subnanometer-sized clusters, reminiscent of Posner clusters, which are stabilized by fetuin-A monomers. Hence, our experiments revealed a second mechanism of long-term mineral ion stabilization by the fetuin-A that is complementary to the formation of CPPs.

Nucleation and growth of CaCO3 mediated by the egg-white protein ovalbumin: a time-resolved in situ study using small-angle neutron scattering.

Mineralization of calcium carbonate in aqueous solutions starting from its initiation was studied by time-resolved small-angle neutron scattering (SANS). SANS revealed that homogeneous crystallization of CaCO 3 involves an initial formation of thin plate-shaped nuclei which subsequently reassemble to 3-dimensional particles, first of fractal and finally of compact structure. The presence of the egg-white protein ovalbumin leads to a different progression of mineralization through several stages; the first step represents amorphous CaCO 3, whereas the other phases are crystalline. The formation and dissolution of the amorphous phase is accompanied by Ca (2+)-mediated unfolding and cross-linking of about 50 protein monomers showing the characteristic scattering of linear chains with a large statistical segment length. The protein complexes act as nucleation centers for the amorphous phase because of their enrichment by Ca (2+) ions. SANS revealed the sequential formation of CaCO 3 starting from the amorphous phase and the subsequent formation of the crystalline polymorphs vaterite and aragonite. This formation from less dense to more dense polymorphs follows the Ostwald-Volmer rule.

Core-shell structure of degradable, thermosensitive polymeric micelles studied by small-angle neutron scattering.

The structure of assemblies of block copolymers composed of thermosensitive, biodegradable poly(N-(2-hydroxypropyl) methacrylamide-dilactate) and poly(ethylene glycol) (pHPMAmDL-b-PEG) has been studied by small-angle neutron scattering (SANS). Three amphiphilic copolymers with a fixed PEG of 5 kDa and a partially deuterated pHPMAmDL(d) block of 6700, 10400, or 21200 Da were used to form micelles in aqueous media by heating the polymeric solution from below to above the cloud point temperature (around 10 degrees C) of the thermosensitive block. Simultaneous and quantitative analysis of the scattering cross sections obtained at three different solvent contrasts is expedited using core-shell model, which assumed a homogeneous core of uniform scattering length density. The mean core radius increased from 13 to 18.5 nm with the molecular weight of the pHPMAmDL(d) block, while the thickness of the stabilizing PEG layer was around 8 nm for the three investigated assemblies. In addition, the volume fraction values of the stabilizing PEG chains in the shell are low and decreased from 31% to 14% with increasing the size of pHPMAmDL(d) block which shows that the shell layer of the assemblies is highly hydrated. The corresponding PEG chain grafting densities decreased from 0.22 to 0.11 nm-2 and the distance between PEG chains on the nanoparticles surface increased from 2.4 to 3.4 nm. The pHPMAmDL-b-PEG micelles showed a controlled instability due to hydrolysis of the lactic acid side groups in the thermosensitive block; that is, an increase of the degradation time leads to an increase of the size of the core which becomes less hydrophobic and consequently more hydrated. Neutron experiments supplied accurate information on how the size of the core and the micelle's aggregation number changed with the incubation time. This feature and the initially small size and dense structure in aqueous solution make the polymeric micelles suitable as carriers for hydrophobic drugs.

The A-B diblock copolymer as a nonordering external field in a three-component A/B/A-B polymer blend.

Thermal copolymer fluctuations were explored in a three-component blend consisting of a critical (A/B) homopolymer blend and a symmetric A-B diblock copolymer using the technique of neutron small angle scattering. The copolymer has the function of an external nonordering field and thereby determines phase behavior as well as the regimes of 3d-Ising, isotropic Lifshitz, and Brasovskiî critical universality. It was found that the random phase approximation (RPA) does not correctly describe the copolymer structure function because of strong thermal fluctuations. On the other hand a weak coupling of copolymer and homopolymer was confirmed, consistent with predictions from RPA. Self-assembly of the copolymers was observed prior to the ordering of the "total" blend, e.g. inclusive of the homopolymers, into bicontinuous and lamellar ordered phases.

Small-angle neutron scattering characterization of polyhydroxyalkanoates and their BioPEGylated hybrids in solution.

Small-angle neutron scattering was used to probe the molecular conformation of various polyhydroxyalkanoates (PHAs) and their bioPEGylated counterparts (PHA- b-PEG). Analysis of neutron scattering profiles of these polymers dissolved in deuterated chloroform at various concentrations from dilute (approximately 0.1% w/v) to semidilute (approximately 7% w/v) showed the two distinct regimes and established overlap concentrations around 4-9 mg mL(-1). Scattering profiles were similar for all polymers investigated; power laws of approximately Q(-1.66) at high Q demonstrated that chloroform behaves as a good solvent for PHAs and suggests that under conditions synonymous with processing the solvated chains were swollen rather than in Gaussian conformation as previously reported. A gradual change to Guinier knees was followed by slopes of Q(-3) suggesting the presence of supramolecular structures at larger length scales. These observations in both the dilute and semidilute concentrations have not been previously reported. Zimm analysis of the data provided gyration radii and absolute molecular weights consistent with trends established using light scattering but showed some variation in their second virial coefficients. While natural-synthetic hybrids of PHA- b-PEG can self-assemble into microporous films, they showed no noticeable differences in chain conformation when in solution, the fabricating medium. This suggests that some form of entropic inducement is required.

Dynamic flexibility of double-stranded RNA activated PKR in solution.

PKR, an interferon-induced double-stranded RNA activated serine-threonine kinase, is a component of signal transduction pathways mediating cell growth control and responses to stress and viral infection. Analysis of separate PKR functional domains by NMR and X-ray crystallography has revealed details of PKR RNA binding domains and kinase domain, respectively. Here, we report the structural characteristics, calculated from biochemical and neutron scattering data, of a native PKR fraction with a high level of autophosphorylation and constitutive kinase activity. The experiments reveal association of the protein monomer into dimers and tetramers, in the absence of double-stranded RNA or other activators. Low-resolution structures of the association states were obtained from the large angle neutron scattering data and reveal the relative orientation of all protein domains in the activated kinase dimer. Low-resolution structures were also obtained for a PKR tetramer-monoclonal antibody complex. Taken together, this information leads to a new model for the structure of the functioning unit of the enzyme, highlights the flexibility of PKR and sheds light on the mechanism of PKR activation. The results of this study emphasize the usefulness of low-resolution structural studies in solution on large flexible multiple domain proteins.

Ginzburg number of a homopolymer-diblock copolymer mixture covering the 3D-Ising, isotropic Lifshitz, and Brasovskii classes of critical universality.

The Ginzburg number Gi of deuterated poly(butadiene) (dPB) and poly(styrene) (PS) homopolymer blend of critical composition mixed with a dPB-PS symmetric diblock copolymer was determined from small angle neutron scattering. A 3 orders of magnitude change of Gi was determined between binary polymer blend and diblock copolymer melt. The strongest change of Gi is observed within the isotropic Lifshitz regime of critical universality occurring over a 3% range of diblock concentration and interpolates the corresponding Gi of the 3D-Ising and Brasovskii regimes. A Lifshitz critical point was not observed consistent with the proposed lower critical dimension d(LCP)=4.