Systematic Quantification of Electron Transfer in a Bare Phospholipid Membrane Using Nitroxide-Labeled Stearic Acids: Distance Dependence, Kinetics, and Activation Parameters.

In this report, we present a method to characterize the kinetics of electron transfer across the bilayer of a unilamellar liposome composed of 1,2-dimyristoyl-sn-glycero-3-phosphocholine. The method utilizes synthetic phospholipids containing noninvasive nitroxide spin labels having the >N-O• moiety at well-defined distances from the outer surface of the liposome to serve as reporters for their local environment and, at the same time, permit measurement of the kinetics of electron transfer. We used 5-doxyl and 16-doxyl stearic acids. The paramagnetic >N-O• moiety is photo-oxidized to the corresponding diamagnetic oxoammonium cation by a ruthenium electron acceptor formed in the solution. Electron transfer is monitored by three independent spectroscopic methods: by both steady-state and time-resolved electron paramagnetic resonance and by optical spectroscopy. These techniques allowed us to differentiate between the electron transfer rates of nitroxides located in the outer leaflet of the phospholipid bilayer and of those located in the inner leaflet. Measurement of electron transfer rates as a function of temperature revealed a low-activation barrier (ΔG‡ ∼ 40 kJ/mol) that supports a tunneling mechanism.

Amino Acid Induced Modification of Self-Assembled Monoglyceride-Based Nanostructures.

Self-assembled phases based on monoglycerides are promising candidates for drug delivery systems. Alterations of these phases need to be performed by addition of substances which are biocompatible. Inverse bicontinuous cubic phases are altered by the addition of five amino acids, namely, glycine, phenylalanine, alanine, glutamine, and tryptophan. These natural molecules have a diversity of side chains which predicts their polarity and subsequently their interaction with the interfacial region. Whereas polar amino acids cause a slight shrinking of the fully hydrated phase, amino acids with a nonpolar side chain expand it. Tryptophan is also able to provoke a growth of inverse hexagonal, micellar cubic, and micellar structures. Amino acid concentrations in the aqueous phase, even above the amino acid's solubility, further affect all aforementioned structures and cause a significant enlargement of up to 26%. Besides the amino acids' impact on the structural sizes, they also affect the phase transition temperatures.

Dopamine-Assisted Modification of Polypropylene Film to Attain Hydrophilic Mineral-Rich Surfaces.

The presented study focuses on the modification of polypropylene (PP) film with tetraethyl orthosilicate (TEOS) under heterogeneous conditions via polydopamine/polyethylene imine (PDA/PEI) chemistry using a facile dip-coating procedure to attain hydrophilic mineral-rich surfaces. Thus, the resulting PP-based films were further immersed in ion-rich simulated body fluid (SBF) to deposit Ca-based minerals onto the film's surfaces efficiently. In addition, the chemical reaction mechanism on PP film was proposed, and mineralisation potential inspected by determination of functional groups of deposits, zeta potential, hydrophilicity and surface morphology/topography using Fourier transform infrared (FTIR) spectroscopy, streaming potential, water contact angle (WCA), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The obtained results show the improved wettability of samples on account of PDA inclusion (WCA was reduced from 103° for pure PP film to 28° for PDA-modified film), as well as the presence of functional groups, due to the PDA/PEI/TEOS surface functionalisation, increased the ability of minerals to nucleate on the PP film's surface when it was exposed to an SBF medium. Moreover, the higher surface roughness due to the silica coatings influenced the enhanced anchoring and attachment of calcium phosphate (CaP), revealing the potential of such a facile approach to modify the chemically inert PP films, being of particular interest in different fields, including regenerative medicine.

Optimized loading and sustained release of hydrophilic proteins from internally nanostructured particles.

In this study, we demonstrate that emulsified microemulsions and micellar cubosomes are suitable as sustained delivery vehicles for water-soluble proteins. Through structural modifications, the loading efficiency of two model proteins, namely bovine serum albumin (BSA) and cytochrome c could be remarkably increased. A procedure for preparing these particles loaded with optimized amounts of sensitive substances is presented. Loading and dispersion at low temperatures is performed in two successive steps. First, a water-in-oil microemulsion is loaded with the proteins. Subsequently, this phase is dispersed in water resulting in particles with microemulsion and micellar cubic internal structure and a size of approximately 620 nm. This two-step method ensures optimal loading of the particles with the proteins. These nanostructured particles are able to sustain the release of the water-soluble BSA and cytochrome c. Within one day, less than 10% of BSA and 15% of cytochrome c are released. The release rate of cytochrome c is influenced by the nanostructure of the particles.

Inverse ISAsomes in Bio-Compatible Oils-Exploring Formulations in Squalane, Triolein and Olive Oil.

In contrast to their more common counterparts in aqueous solutions, inverse ISAsomes (internally self-assembled somes/particles) are formulated as kinetically stabilised dispersions of hydrophilic, lyotropic liquid-crystalline (LC) phases in non-polar oils. This contribution reports on their formation in bio-compatible oils. We found that it is possible to create inverse hexosomes, inverse micellar cubosomes (Fd3m) and an inverse emulsified microemulsion (EME) in excess squalane with a polyethylene glycol alkyl ether as the primary surfactant forming the LC phase and to stabilise them with hydrophobised silica nanoparticles. Furthermore, an emulsified L1-phase and inverse hexosomes were formed in excess triolein with the triblock-copolymer Pluronic® P94 as the primary surfactant. Stabilisation was achieved with a molecular stabiliser of type polyethylene glycol (PEG)-dipolyhydroxystearate. For the inverse hexosomes in triolein, the possibility of a formulation without any additional stabiliser was explored. It was found that a sufficiently strong stabilisation effect was created by the primary surfactant alone. Finally, triolein was replaced with olive oil which also led to the successful formation of inverse hexosomes. As far as we know, there exists no previous contribution about inverse ISAsomes in complex oils such as triolein or plant oils, and the existence of stabiliser-free (i.e., self-stabilising) inverse hexosomes has also not been reported until now.

Influence of steam jet-cooking on the rheological properties of dry and wet cationized starch solutions.

Steam jet-cooking allows for efficient dissolution of cationic starch in paper production as it operates above the boiling point of water at elevated pressures. However, the processes involved during jet-cooking and its consequences on dissolution and finally paper properties have not been fully resolved so far. As cationic starch is the most important paper additive in the wet end, any energy or material savings during dissolution will enhance the ecologic and economic performance of a paper mill. Here, we address the topic of solubilization of four different industrially relevant cationic starches processed via steam jet-cooking. We showcase that rheology is a useful tool to assess the solubility state of starches. Some starches featured liquid-like rheological behavior (loss moduli, G", greater than storage moduli, G') in linear viscoelastic tests and anti-thixotropic behavior in hysteresis loop tests. In contrast, cationic corn starches exhibited gel-like behavior (G' > G″) and negligible hysteresis directly after cooking. HYPOTHESES: To evaluate the decisive factors for complete dissolution of industrial cationic starches using jet-cooking and to correlate them to rheological properties.

The metastasis-associated extracellular matrix protein osteopontin forms transient structure in ligand interaction sites.

Osteopontin (OPN) is an acidic hydrophilic glycophosphoprotein that was first identified as a major sialoprotein in bones. It functions as a cell attachment protein displaying a RGD cell adhesion sequence and as a cytokine that signals through integrin and CD44 cell adhesion molecules. OPN is also implicated in human tumor progression and cell invasion. OPN has intrinsic transforming activity, and elevated OPN levels promote metastasis. OPN gene expression is also strongly activated in avian fibroblasts simultaneously transformed by the v-myc and v-mil(raf) oncogenes. Here we have investigated the solution structure of a 220-amino acid recombinant OPN protein by an integrated structural biology approach employing bioinformatic sequence analysis, multidimensional nuclear magnetic resonance spectroscopy, synchrotron radiation circular dichroism spectroscopy, and small-angle X-ray scattering. These studies suggest that OPN is an intrinsically unstructured protein in solution. Although OPN does not fold into a single defined structure, its conformational flexibility significantly deviates from random coil-like behavior. OPN comprises distinct local secondary structure elements with reduced conformational flexibility and substantially populates a compact subspace displaying distinct tertiary contacts. These compacted regions of OPN encompass the binding sites for α(V)ß(III) integrin and heparin. The conformational flexibility combined with the modular architecture of OPN may represent an important structural prerequisite for its functional diversity.

Fetal HDL/apoE: a novel regulator of gene expression in human placental endothelial cells.

Maternal lipoproteins have been studied extensively in human pregnancies, but little is known about the role of fetal lipoproteins. The vascularized human placenta interfaces between the mother and fetus to transfer nutrients for sustaining pregnancy. Unlike that of adults, fetal high-density lipoprotein (HDL), which is in contact with placental vessels, is characterized by a high proportion of apolipoprotein E (apoE). We hypothesize this unique composition of fetal HDL affects key functions of the growing fetal tissues. The aim was to identify genes regulated by apoE-HDL by incubating human placental endothelial cells (HPEC) with either fetal HDL or apoE-rich reconstituted HDL particles (apoE-rHDL). HPEC were exposed to 15 µg/ml fetal HDL, 15 µg/ml apoE-rHDL, or medium for 16 h, respectively. Microarray analysis determined genes regulated by fetal HDL and apoE. Characterization of HDL particles revealed a different hydrodynamic radius for apoE-rHDL (13.70 nm) compared with fetal HDL (18.11 nm). Stepwise gene clustering after microarray experiments identified 79 differentially expressed genes (P < 0.05) when cells were exposed to HDL compared with controls. Among them 16 genes were downregulated, whereas five genes were upregulated by twofold, respectively. When HPEC were incubated with apoE-rHDL 18-fold more genes (1,417, 12% of transcripts) were regulated (P < 0.05) in contrast to HDL. Thereof, 172 genes were downregulated and 376 genes upregulated (twofold). In the common subset of 38 genes regulated by both HDL particles, genes involved in cholesterol biosynthesis and cell protection prevailed. Strikingly, results suggest that HDL has the capability of regulating metallothioneins, which may have an effect on oxidative stress in HPEC.

High-Yield Production of Selected 2D Materials by Understanding Their Sonication-Assisted Liquid-Phase Exfoliation.

Liquid-phase exfoliation (LPE) is a widely used and promising method for the production of 2D nanomaterials because it can be scaled up relatively easily. Nevertheless, the yields achieved by this process are still low, ranging between 2% and 5%, which makes the large-scale production of these materials difficult. In this report, we investigate the cause of these low yields by examining the sonication-assisted LPE of graphene, boron nitride nanosheets (BNNSs), and molybdenum disulfide nanosheets (MoS2 NS). Our results show that the low yields are caused by an equilibrium that is formed between the exfoliated nanosheets and the flocculated ones during the sonication process. This study provides an understanding of this behaviour, which prevents further exfoliation of nanosheets. By avoiding this equilibrium, we were able to increase the total yields of graphene, BNNSs, and MoS2 NS up to 14%, 44%, and 29%, respectively. Here, we demonstrate a modified LPE process that leads to the high-yield production of 2D nanomaterials.

Influence of the stabilizer concentration on the internal liquid crystalline order and the size of oil-loaded monolinolein-based dispersions.

The internal phase of monolinolein-based dispersions loaded with tetradecane or (R)-(+)-limonene was investigated as a function of the stabilizer content by small-angle X-ray scattering. Phase transitions at the colloidal scale were found in some of nanostructured aqueous dispersions by increasing the stabilizer content. For particles containing a bicontinuous cubic phase, a large increase of the stabilizer concentration promoted a liquid crystalline phase transition from the Pn3m to the Im3m cubic symmetry. The coexistence of both phases is observed in an intermediate stabilizer concentration range. For particles with an internal micellar cubic Fd3m symmetry, the internal structure changes in the isotropic fluid L(2) phase. In case of particles with an internal hexagonal phase (H(2) symmetry), the increasing amount of stabilizer did not alter the lattice parameter but decreased the size of the nanostructured domain. Moreover, we showed for hexagonal and emulsified micellar phase particles that the increase of the stabilizer content induced a strong decrease of the mean hydrodynamic size of the particles, allowing producing nanostructured lipid-based liquid crystalline particles down to a radius of 70 nm at the same energy input.

Separation and purification of nanoparticles in a single step.

Reversed-micelle synthesis has been used to generate CTAB-stabilized gold (Au-NPs) and silver nanoparticles (Ag-NPs). By inducing a phase transition and subsequent separation of the background supporting microemulsion, it has been possible to extract and purify the NPs from the reaction medium. After addition of excess water, the NPs concentrate into an upper octane-rich phase, with impurities and reaction debris (in particular CTAB) partitioning into the water-rich lower phase. UV and (1)H NMR showed that 82% of the original mass of Au-NPs can be purified from the excess CTAB and other salt impurities. The concentrated and purified NPs can be dried down, by solvent removal, and then redispersed in octane. Using the complementary techniques small-angle neutron and X-ray scattering (SANS and SAXS), the structures of microemulsions both with and without nanoparticles prior to separation, and in both upper and lower phases after separation, have been elucidated. The approach has also been applied to the synthesis and recovery of silver nanoparticles, but on a larger scale. This new approach compares favorably with existing methods as it uses no additional organic solvents, has a low-energy demand, and requires no specialist surfactants. The new advance here is that by using a colloidal system to prepare and support the nanoparticles as a structured solvent, a simple soft purification method becomes accessible, which is otherwise impossible with a normal molecular solvent.

Vancomycin Loaded Glycerol Monooleate Liquid Crystalline Phases Modified with Surfactants.

The influence of two tuning agents, polyglycerol ester (PE) and triblock copolymer (TC), on the properties of glycerol monooleate (MO) liquid crystalline phase (LCP) was investigated to achieve the therapeutic concentration of vancomycin hydrochloride (VHCl) into the eye, topically during 60 min (1 h) and intravitreally during 2880 min (48 h). Different techniques were used to elucidate the impact of surfactants on the structure of the LCP: polarized light microscopy (PLM), small-angle X-ray scattering (SAXS), and in vitro release tests I and II (simulating local and intravitreal application in the eye). The structure analysis by SAXS depicts that the inclusion of PE into the MO LCP provided partial transition of a hexagonal phase into a lamellar phase, and TC induced a partial transition of a hexagonal phase into an LCP which identification was difficult. The LCP modulated with PE and TC demonstrated different VHCl's release patterns and were evaluated by comparing our release data with the literature data. The comparison indicated that the LCP modulated with 30% w/w PE could be a promising VHCl delivery system intravitreally during 2880 min.

Cationic starches in paper-based applications-A review on analytical methods.

This review focuses on cationic starches with a low degree of substitution (<0.06) which are mainly used for production of paper-based products. After a brief introduction on starch in general, cationization pathways and importance of cationic starches in paper production, this review emphasizes on the analytical challenges from different perspectives. These include the different length scales of starches when in solution: the macromolecular level, their assembly into nm aggregates and finally hydrocolloids with hundreds of nanometers of diameter. We give an overview on the current state of the art on the analysis of such challenging samples and aim at providing a guideline for obtaining and presenting reliable analytical data.

Investigation of the Adsorption Behavior of Jet-Cooked Cationic Starches on Pulp Fibers.

The optimization of the thermal treatment of cationic starch in the paper industry offers the opportunity to reduce the energy consumption of this process. Four different industrially relevant cationic starches, varying in source, cationization method and degree of substitution were treated by a steam-jet cooking procedure, comparable to industrially employed starch cooking processes. The influence of the starch properties and cooking parameters on the adsorption behavior of the starches on cellulosic pulp was investigated. The adsorbed amount was affected by the cooking temperature and the type of starch. For some starch grades, a cooking temperature of 115 °C can be employed to achieve sufficient starch retention on the pulp fibers. The energy consumption could further be reduced by cooking at higher starch concentrations without loss of adsorption efficiency.

Ordered structures in carboxymethylcellulose-cationic surfactants-copper ions precipitated phases: in situ formation of copper nanoparticles.

Small-angle X-ray scattering has been used to investigate the nanostructures in precipitated phases of carboxymethylcellulose/oppositely charged C(n)TAB surfactants mixtures in interaction with bivalent copper ions. In copper-free precipitates, a transition from a 2D hexagonal phase H(I) to a micellar cubic surfactant structure Pm3n was found on increasing the polymer concentration (n = 14, 16). By addition of small amounts of copper ions, the internal structure also changes from H(I) to Pm3n cubic and finally turns into a less-ordered phase. The concentration at which this transition occurs strongly depends on the surfactant tail length. The presence of copper ions in the precipitated phase leads at the same time to a disorganization of the surfactant micelles' order and to the formation of a lamellar ordering, revealed by the presence of additional diffraction peaks above 10 mM copper. We used the polymer/surfactant networks as template systems, controlled by the copper ion content, for preparing embedded copper nanoparticles via in situ reduction of copper sulfate by sodium borohydride. The zerovalent copper particles formed in the cubic lattice are well-defined in size (less than 10 nm) and have a homogeneous distribution, whereas they are very polydisperse when the reduction is performed in the unorganized phase because the lack of order in the matrix does not protect them from aggregation.

Vancomycin ocular delivery systems based on glycerol monooleate reversed hexagonal and reversed cubic liquid crystalline phases.

Lyotropic bulk reversed hexagonal and reversed cubic liquid crystalline phases (hexagonal and cubic phases) composed of glycerol monooleate (GM) were used to design the vancomycin hydrochloride's (VHCl) delivery systems aiming to maintain VHCl's therapeutic concentration during 24 h in the eye, locally (as an insert) and/or intravitreally (as a bulk phase injection). Bulk VHCl's hexagonal and cubic phases were successfully prepared by melted homogenization and solvent evaporation method, and then an insert was prepared. The structural characteristics of liquid crystalline phases were studied using cross polarized light microscopy and small angle X-ray scattering technique. The presence of VHCl (1-9.5% w/w VHCl solution) did not exhibit any change in the liquid crystalline phase's structure to another liquid crystalline phase, and showed little effect on the lattice parameter of the existing liquid crystalline phase structure. In order to relate the liquid crystalline phase structure to VHCl's release rate locally into the eye, in-vitro release test of an implant has been done using a simulated tear fluid. VHCl's release in the simulated tear fluid from the cubic phase obeyed Higuchi kinetics, with linear VHCl's release versus the square root of time. The hexagonal phase released VHCl in simulated tear fluid significantly slower than the cubic phase. In order to relate the liquid phase structure to VHCl's diffusion intravitreally, in vitro release test by the Sirius' Subcutaneous Injection Site Simulator (Scissor) has been performed. Comparing the release properties by a Scissor, the VHCl's cubic phase demonstrated slower permeation through extra-cellular matrix than the VHCl solution. To evaluate the efficacy of the system investigated, the release properties of VHCl's cubic phase were compared with literature data indicating that the cubic phase could be a potential matrix system in the delivery of VHCl intravitreally during 24 h after intravitreal injection. The release data in the simulated tear fluid indicated that the cubic phase should be further optimized to achieve a therapeutic VHCl concentration locally in the eye during 24 h.

Interaction of industrially relevant cationic starches with cellulose.

Industrially relevant, commercially available cationic starches have been investigated towards their interaction capacity with cellulose thin films derived from trimethylsilyl cellulose (TMSC). The starches used in this study stem from different sources (potato, pea, corn) and featured rather low degrees of substitution ranging from 0.030 to 0.062. The interaction of those starches with cellulose thin films was studied by surface plasmon resonance spectroscopy under flow conditions using concentrations of 1.0mgml-1 and a flow rate of 25µlmin-1. All the investigated starches employed in this study were capable to efficiently interact with the slightly negatively charged cellulose surface leading to irreversible deposition on the surface. As complementary techniques atomic force microscopy and x-ray photoelectron spectroscopy were used to confirm the presence of the starches on the cellulose film surface. Further, dynamic light scattering and size exclusion chromatography measurements were performed to correlate adsorbed amount, particle size and molecular weight of the starches to their interaction behavior.

Design of simultaneous antimicrobial and anticoagulant surfaces based on nanoparticles and polysaccharides.

The rational design of silver nanoparticles encapsulated in an anticoagulant, hemocompatible polysaccharide, 6-O-chitosan sulfate, is presented. Three different approaches are described for the immobilization of these core shell particles on cellulosic surfaces. The mass of the immobilized particles is quantified using a quartz crystal microbalance with dissipation (QCM-D). The antimicrobial activity of the surfaces towards E. coli MG 1655 [R1-16] is investigated by live/dead assays using fluorescence staining. All surfaces treated with the designed nanoparticles exhibit excellent antimicrobial activity towards E. coli MG 1655 [R1-16]. Anticoagulant properties of blood plasma on the nanoparticle treated surfaces have been determined using QCM-D. In comparison with the unmodified substrates, the total coagulation time as well as the thrombin formation time and fibrin clotting time of surfaces modified with nanoparticles are significantly increased.