During embryonic development, the proepicardial organ (PEO) grows out over the heart surface to form the epicardium. Following epithelial-mesenchymal transformation, epicardium-derived cells (EPDCs) migrate into the heart and contribute to the developing coronary arteries, to the valves, and to the myocardium. The peripheral Purkinje fiber network develops from differentiating cardiomyocytes in the ventricular myocardium. Intrigued by the close spatial relationship between the final destinations of migrating EPDCs and Purkinje fiber differentiation in the avian heart, that is, surrounding the coronary arteries and at subendocardial sites, we investigated whether inhibition of epicardial outgrowth would disturb cardiomyocyte differentiation into Purkinje fibers. To this end, epicardial development was inhibited mechanically with a membrane, or genetically, by suppressing epicardial epithelial-to-mesenchymal transformation with antisense retroviral vectors affecting Ets transcription factor levels (n=4, HH39-41). In both epicardial inhibition models, we evaluated Purkinje fiber development by EAP-300 immunohistochemistry and found that restraints on EPDC development resulted in morphologically aberrant differentiation of Purkinje fibers. Purkinje fiber hypoplasia was observed both periarterially and at subendocardial positions. Furthermore, the cells were morphologically abnormal and not aligned in orderly Purkinje fibers. We conclude that EPDCs are instrumental in Purkinje fiber differentiation, and we hypothesize that they cooperate directly with endothelial and endocardial cells in the development of the peripheral conduction system.

The secret of a successful affinity biosensor partially hides in the chemical interface layer between the transducer system and the biological receptor molecules. Over the past decade, several methodologies for the construction of such interface layers have been developed on the basis of the deposition of self-assembled monolayers (SAMs) of alkanethiols on gold. Moreover, mixed SAMs of polyethylene oxide (PEO) containing thiols have been applied for the immobilization of biological receptors. Despite the intense research in the field of thiol SAMs, relatively little is known about their biosensing properties in correlation with their long-term stability. Especially the impact of the storage conditions on their biosensing characteristics has not been reported before to our knowledge. To address these issues, we prepared mixed PEO SAMs and tested their stability and biosensing performance in several storage conditions, i.e., air, N2, ethanol, phosphate buffer, and H2O. The quality of the SAMs was monitored as a function of time using various characterization techniques such as cyclic voltammetry, contact angle, grazing angle Fourier-transform infrared spectroscopy, and X-ray photoelectron spectroscopy. In addition, the impact of the different storage conditions on the biosensor properties was investigated using surface plasmon resonance. Via the latter technique, the receptor immobilization, the analyte recognition, and the nonspecific binding were extensively studied using the prostate specific antigen as a model system. Our experiments showed that very small structural differences in the SAM can have a great impact in their final biosensing properties. In addition it was shown that the mixed SAMs stored in air or N2 are very stable and retain their biosensor properties for at least 30 days, while ethanol appeared to be the worst storage medium due to partial oxidation of the thiol headgroup. In conclusion, care must be taken to avoid SAM degradation during storage to retain typical SAM characteristics, which is very important for their general use in many proposed applications.

In the present work, sustained release gastroretentive minimatrices of amoxicillin have been designed and optimized using central composite design. Effect of amount of xanthan gum, rate controlling polymers (HPMC K100M CR/PEO coagulant (1:1)), carbopol 974P, and gas generating couple (sodium bicarbonate/citric acid (3:1)) was studied on dependent (response) variables, i.e., buoyancy lag time, drug release at 1 h, time required for 95% drug release, swelling index, and bioadhesive strength. Minimatrices were prepared by non aqueous granulation method using solution of PVP K30 in isopropyl alcohol. All the formulations were found to contain 99.2% to 100.9% of amoxicillin per minimatrix. Optimum formulation (Formulation number AGT09) containing high level of the independent variables was having buoyancy lag time of 7 min and drug release at 1 h was 32.5%. It required 9.39 h for 95% drug release while swelling index and bioadhesive strength were 341 and 17.9 dyn/cm(2), respectively. This formulation was said to be optimum because it has minimum buoyancy lag time, requires maximum time for 95% drug release, and has higher bioadhesive capabilities. In vitro results of an optimized formulation indicate its sustained drug release and gastric retention capability, which may be very useful for effective treatment of H. pylori infection.

A composite gastro-retentive matrix for zero-order delivery of highly soluble drug alfuzosin hydrochloride (10mg) has been designed and characterized. Two systems containing polyethylene oxide (PEO), hydroxypropylmethylcellulose (HPMC), sodium bicarbonate, citric acid and polyvinyl pyrrolidone were dry blended and compressed into triple layer and bi-layer composite matrices. Dissolution studies using the USP 27 paddle method at 100 and 50rpm in pH 2.0 and 6.8 were performed using UV spectroscopy at 244nm, with automatic sampling over a 24h period using a marketed product as a reference to calculate the "f(2)" factor. Textural characteristics of each layer, the composite matrix as a whole, and floatation potential were determined under conditions similar to dissolution. Percent matrix swelling and erosion along with digital images were also obtained. Both systems proved to be effective in providing prolonged floatation, zero-order release, and complete disentanglement and erosion based on the analysis of data with "f(2)" of 68 and 71 for PEO and HPMC based systems, respectively. The kinetics of drug release, swelling and erosion, and dynamics of textural changes during dissolution for the designed composite systems offer a novel approach for developing gastro-retentive drug delivery system that has potential to enhance bioavailability and site-specific delivery to the proximal small intestine.

Cell functions are regulated by signal transduction processes triggered by binding interactions with extracellular ligands, including those coupled to insoluble matrices as well as those diffusing in free solution. Whereas receptor interactions with freely diffusible soluble ligands are quantitatively governed by the mean ligand concentration, interactions with spatially constrained substratum-bound ligands may be affected not only by the mean ligand concentration, but also by the ligand spatial microdistribution. To probe this hypothesis we have generated surfaces presenting galactose ligands at different concentrations and different capabilities for micromobility--and thus spatial microdistribution--by means of polyethylene oxide tethering and assayed for hepatocyte spreading mediated by asialoglycoprotein receptor/cytoskeleton linkages. We demonstrate that spreading is not uniquely determined by the mean galactose concentration presented by the culture substrate. Rather, the ability of primary hepatocytes to spread is additionally specified by a combination of ligand concentration and tether length. Our results suggest spreading results when monovalent ligands possess sufficient mobility to form effectively multivalent bonds with the receptor; that is, when the substratum-bound ligands to cluster in spatial microdomains.

We have performed atomistic molecular dynamics simulations of an anionic sodium dodecyl sulfate (SDS) micelle and a nonionic poly(ethylene oxide) (PEO) polymer in aqueous solution. The micelle consisted of 60 surfactant molecules, and the polymer chain lengths varied from 20 to 40 monomers. The force field parameters for PEO were adjusted by using 1,2-dimethoxymethane (DME) as a model compound and matching its hydration enthalpy and conformational behavior to experiment. Excellent agreement with previous experimental and simulation work was obtained through these modifications. The simulated scaling behavior of the PEO radius of gyration was also in close agreement with experimental results. The SDS-PEO simulations show that the polymer resides on the micelle surface and at the hydrocarbon-water interface, leading to a selective reduction in the hydrophobic contribution to the solvent-accessible surface area of the micelle. The association is mainly driven by hydrophobic interactions between the polymer and surfactant tails, while the interaction between the polymer and sulfate headgroups on the micelle surface is weak. The 40-monomer chain is mostly wrapped around the micelle, and nearly 90% of the monomers are adsorbed at low PEO concentration. Simulations were also performed with multiple 20-monomer chains, and gradual addition of polymer indicates that about 120 monomers are required to saturate the micelle surface. The stoichiometry of the resulting complex is in close agreement with experimental results, and the commonly accepted "beaded necklace" structure of the SDS-PEO complex is recovered by our simulations.

The influence of various excipients on the conversion of carbamazepine polymorphs to the dihydrate in aqueous suspension has been investigated. Ten excipients having functional groups which were potentially able to form hydrogen bonds with carbamazepine (group 1: methylcellulose, hypromellose (hydroxypropyl methylcellulose), hydroxypropylcellulose (HPC), 2-hydroxyethylcellulose (HEC), carmellose sodium (sodium carboxymethylcellulose), cellobiose; group 2: povidone (polyvinylpyrrolidone), povidone-vinyl acetate copolymer (povidone/VA) and N-methyl-2-pyrrolidone; group 3: macrogol (polyethylene glycol) and polyethylene oxide-polypropylene oxide copolymer (PEO/PPO)) were selected. Carbamazepine polymorphic forms III and I were dispersed separately into each aqueous excipient solution (0.1%, w/v) for 30 min at room temperature. The inhibition effect of each excipient was quantified using Raman spectroscopy combined with multivariate analyses. The solubility parameter of each excipient was calculated and used for categorizing excipients. Excipients in groups 1 and 2, which had both low solubility parameters (< 27.0 MPa(1/2)) and strong hydrogen bonding groups, inhibited the conversion completely. With increasing solubility parameter, the inhibition effect decreased for group 1 excipients, especially for carbamazepine form I, which had a higher specific surface area. Also, the excipients of group 3, lacking strong hydrogen bonding groups, showed poor inhibition although they had low solubility parameters (< 21.0 MPa(1/2)). This study indicated the importance of both hydrogen bonding interaction and a suitable hydrophobicity (expressed by the solubility parameter) in the inhibition of the conversion of carbamazepine to the dihydrate.

A stable drug carrier has been prepared by covalently coating magnetic nanoparticles (MNPs) with PEO-PPO-PEO block copolymer Pluronic P85. The particles were characterized by TEM, XRD, DLS, VSM, FTIR, and TGA. A typical product has a 15 nm magnetite core and a 100 nm hydrodynamic diameter with a narrow size distribution and is superparamagnetic with large saturation magnetization (57.102 emu/g) at room temperature. The covalently-coated Pluronic-MNPs (MagPluronics) were proven to be stable in different conditions, such as aqueous solution, 0.2 M PBS solution, and pH 13.5 solution, which would be significant for biological applications. Furthermore, MagPluronics also possess temperature-responsive property acquired from the Pluronic copolymer layer on their surface, which can cause conformational change of Pluronics and improve load and delivery efficiency of the particles. The temperature-controlled loading and releasing of hydrophobic model drug curcumin were tested with these particles. A loading efficiency of 81.3% and a sustained release of more than 4 days were achieved in simulated human body condition. It indicates that the covalently-coated MagPluronics are stable carriers with good drug-loading capacity and controlled-release property.

OBJECTIVE

To investigate the relative aggregation state of amphotericin B (AmB) during loading and reconstitution of polymeric micelles.

METHODS

Hexanoate and stearate derivatives of PEO-b-p (L-Asp) were prepared. The polymers and AmB were dissolved in methanol (MeOH). Milli-Q water was then added slowly, and the MeOH was removed via rotary evaporation. The solutions were freeze-dried in the presence of trehalose. During micelle preparation, the aggregation state of AmB was assessed using absorption spectroscopy. Upon reconstitution, the samples were analyzed using vapor-pressure osmometry, size-exclusion chromatography (SEC), and absorption spectroscopy. The absorption spectrum of AmB in the presence of the block copolymers was compared to that of AmB alone under the same conditions.

RESULTS

AmB was loaded into micelles prepared from acyl derivatives of PEO-b-p (L-Asp). Absorption spectroscopy indicated that the aggregation state was preserved during the loading process. AmB exists in a self-aggregated state in polymeric micelles containing hexanoate ester cores and in a relatively monomeric state in polymeric micelles containing stearate ester cores. Vapor-pressure osmometry confirmed the isotonicity of the formulations, while SEC indicated that the micelles were approximately 10(6) g/mol.

CONCLUSIONS

Depending on the polymer structure and assembly conditions, it is possible to encapsulate AmB in a relatively nonaggregated or aggregated state in micelles prepared from acyl derivatives of PEO-b-p (L-Asp). In polymeric micelles containing stearate side chains, AmB was loaded in a nearly monomeric state, possibly due to interaction with the stearate side chains. The final aggregation state of the drug is preserved during lyophilization and reconstitution of polymeric micelles prepared by a novel solvent evaporation procedure.

Energy-filtered transmission electron microscopy (EFTEM) was used to determine the distribution of lithium ions in solid polymer electrolytes for lithium batteries. The electrolytes of interest are mixtures of bis(trifluoromethane)sulfonimide lithium salt and symmetric poly(styrene-block-ethylene oxide) copolymers (SEO). In contrast to current solid and liquid electrolytes, the conductivity of SEO/salt mixtures increases with increasing molecular weight of the copolymers. EFTEM results show that the salt is increasingly localized in the middle of the poly(ethylene oxide) (PEO) lamellae as the molecular weight of the copolymers is increased. Calculations of the inhomogeneous local stress field in block copolymer microdomains, modeled using self-consistent field theory, provide a quantitative explanation for this observation. These stresses, which increase with increasing molecular weight, interfere with the ability of PEO chains to coordinate with lithium cations near the walls of the PEO channels where ion mobility is expected to be low.

We describe the synthesis and in vitro biological characterization of a new class of carbohydrate-modified hydrogels based on radiation-cross-linked star polyethylene oxide (PEO). Hydrogels were synthesized from either of two types of PEO star molecules in order to vary the terminal hydroxyl content of the gels while keeping other gel properties such as molecular weight between cross-links and water content constant. The resulting gels were covalently modified with monosaccharide ligands and the behaviour of primary rat hepatocytes on the modified gels was evaluated under culture conditions. Hepatocytes exhibited a sugar-specific adhesion to the modified gels, adhering to gels bearing galactose but not glucose. Cell spreading was observed on both types of galactose-modified PEO star gels; moreover, the gels supported long-term (6 d) culture and differentiated function of primary hepatocytes. Further, on comparing the cell spreading behaviour observed on the PEO star gels with that reported previously for galactose-modified polyacrylamide, we find that our gels elicit spreading at ligand concentrations lower by an order of magnitude. A simple mechanistic analysis indicates that this enhanced ability of PEO star gels to support spreading of primary hepatocytes on low concentrations of immobilized galactose derives from freedom of the immobilized ligands to come within sufficiently close proximity to mimic a high-affinity branched oligosaccharide.

The shear flow of a triblock copolymer micellar solution (<em>PEO</em>-PPO-<em>PEO</em> Pluronic P84 in brine) is investigated using simultaneous rheological and velocity profile measurements in the concentric cylinder geometry. We focus on two different temperatures below and above the transition temperature T{c} which was previously associated with the apparition of a stress plateau in the flow curve. (i) At T=37.0 degrees C<T{c}, the bulk flow remains homogeneous and Newtonian-like, although significant wall slip is measured at the rotor that can be linked to an inflexion point in the flow curve. (ii) At T=39.4 degrees C>T{c}, the stress plateau is shown to correspond to stationary shear-banded states characterized by two high shear rate bands close to the walls and a very weakly sheared central band, together with large slip velocities at the rotor. In both cases, the high shear branch of the flow curve is characterized by flow instability. Interpretations of wall slip, three-band structure, and instability are proposed in light of recent theoretical models and experiments.

Linear amphiphilic diblock and ternary triblock copolymers were synthesized by the RAFT method in two successive steps using a poly(ethylene oxide) (PEO) macrochain transfer agent, butyl or 2-ethylhexyl acrylate, and 1H,1H,2H,2H-perfluorodecyl acrylate. The diblock and the triblock copolymers, which consist of a hydrophilic, a lipophilic, and a short fluorophilic block, self-assemble in water into spherical micellar aggregates. Imaging by cryogenic transmission electron microscopy (cryo-TEM) revealed that the micellar cores of the aggregates made from these "triphilic" copolymers can undergo local phase separation to form a unique ultrastructure. In these multicompartment micelles, it appears that extended nonspherical domains, presumably made of nanocrystallites of the fluorocarbon block, are embedded in the hydrocarbon matrix forming the spherical micellar core. This novel internal structure of a micellar core is attributed to the mutual incompatibility of the fluorocarbon and hydrocarbon side chains in combination with the tendency of the used fluorocarbon acrylate monomer to undergo side-chain crystallization.

We compared the distribution of deleted mitochondrial DNA (Delta-mtDNA) in skeletal muscle of a patient with autosomal recessive (AR) and another with autosomal dominant (AD) progressive external ophthalmoplegia (PEO) by in situ hybridization (ISH). The patients studied had similar numbers of fibers deficient in cytochrome c oxidase (COX) activity (13.6% and 12.8%) and fibers with mitochondrial proliferation (5.5% and 5.3%). ISH suggested that each COX-deficient fiber contained a single species of Delta-mtDNA. Most deletions ablated the region between the genes encoding adenosine triphosphate (ATP) synthase subunit 8 and cytochrome b. Fibers that appeared to be depleted of mtDNA were also present. We conclude that muscle from patients with autosomally inherited PEO contains not only Delta-mtDNA but also focal depletion of mtDNA and that the distribution of these mtDNA defects appears to be similar. These changes most likely represent the common consequence of whatever genetic factors are responsible for the generation of Delta-mtDNA.

Magnesium (Mg) alloys have been suggested as biodegradable bone implant materials due to their good intrinsic biocompatibility and great mechanical properties. Although magnesium has attractive properties as an orthopedic implant material, its quick degradation and low bioactivity may lead to the loss of mechanical integrity of the implant during the bone healing process. In this paper, we endeavor to surmount the abovementioned defects using the surface coating technique. We have recently coated AZ91 magnesium implants with merwinite (Ca3MgSi2O8) through the coupling of plasma electrolytic oxidation (PEO) and electrophoretic deposition method. In this work, we are specifically focused on the in vivo examinations of the coated implants in comparison with the uncoated one. For the in vivo experiment, the rod samples, including the uncoated and merwinite/PEO coated implants, were imbedded into the greater trochanter of rabbits. The results of the in vivo animal test indicated an improvement in biodegradability including slower implant weight loss, reduction in Mg ion released from the coated implants in the blood plasma, lesser release of hydrogen bubbles and an improvement in biocompatibility including an increase in the amount of bone formation and ultimately a mild bone inflammation after the surgery according to the histological images. In summary, proper surface treatment of magnesium implants such as silicate bioactive ceramics may improve their biocompatibility under physiological conditions to making them suitable and applicable for future clinical applications.

The adsorption and elution of the antimicrobial peptide nisin at silanized silica surfaces coated to present pendant polyethylene oxide chains was detected in situ by zeta potential measurements. Silica microspheres were treated with trichlorovinylsilane to introduce hydrophobic vinyl groups, followed by self assembly of the polyethylene oxide-polypropylene oxide-polyethylene oxide (PEO-PPO-PEO) triblock surfactant Pluronic F108, or an F108 derivative with nitrilotriacetic acid end groups. Triblock-coated microspheres were gamma-irradiated to covalently stabilize the PPO-surface association. PEO layer stability was evaluated by triblock resistance to elution by SDS, and layer uniformity was evaluated by fibrinogen repulsion. Introduction of nisin to uncoated or triblock-coated microspheres produced a significant positive change in surface charge (zeta potential) as a result of adsorption of the cationic peptide. In sequential adsorption experiments, the introduction of fibrinogen to nisin-loaded triblock layers caused a decrease in zeta potential that was consistent with partial elution of nisin and/or preferential location of fibrinogen at the interface. This change was substantially more pronounced for uncoated than triblock-coated silica, indicating that the PEO layer offers enhanced resistance to nisin elution.

The absorption of dyes within hydrogen-bonded and electrostatically assembled multilayers and subsequent release of the dyes from the films were studied in situ using FTIR-ATR. Multilayers were composed of poly(methacrylic acid), PMAA, and poly(ethylene oxide), PEO (hydrogen-bonded multilayers), or of PMAA and 22% quarternized copolymer of N-ethyl-4-vinylpyridium bromide and 4-vinylpyridine, Q22 (electrostatically stabilized multilayers). After multilayer deposition, the solution pH was changed to produce excess charge within the films. Dyes with charge opposite to the excess charge of the film (Rhodamine 6G for hydrogen-bonded films or Bromophenol Blue for electrostatically assembled multilayers) were then allowed to absorb within multilayers. In both systems, the dyes were uniformly included within the films. The top layers largely affected the loading capacity of the multilayers, suggesting weaker binding of the dyes with the top layers. Dye release into a 0.01 M phosphate buffer was significantly smaller as compared to release in the presence of 0.05-0.5 mg/mL solutions of adsorbing polymers whose charge was the same as the excess charge within the films. We found that with the PMAA/PEO films, dye release did not depend on the concentration of polymer in solution, but was largely controlled by the amount of charge accumulated within the adsorbing polymer layer on the top of the film. For electrostatically stabilized PMAA/Q22 systems, dye release increased with increasing concentration of Q22 in solution, suggesting a significant contribution of the competition of solution species in the release mechanism. Our findings contribute to the understanding of interactions of small molecules with polymer multilayers and might have ramifications for novel applications of multilayer films as new materials for the controlled delivery of chemicals.

Poly(ethylene glycol):poly(acrylate) PEG-g-PA co-polymers were made that inhibited nonspecific protein and cellular adhesion. PEG-g-PA co-polymers were then covalently modified with either cell adhesion peptides or fragments of antibodies to monocyte/macrophage integrin receptors (anti-VLA4, anti-beta(1), anti-beta(2), and anti-CD64) known to enhance macrophage adhesion and, perhaps, modulate their activation. Peptides were either directly conjugated to the base material or linked by way of PEO-star tethers. Fragments of the antibody region containing the antigen-binding site (Fab' fragments) were coupled to other PEG-g-PA samples using the sulhydryl end groups on Fab' fragments to amine-bearing PEO stars. Macrophage adhesion rates, phagocytic response (oxidative burst), and cytokine expression were determined for each PEG-g-PA material. Luminol-enhanced chemiluminescence was used as a semiquantitative indication of monocyte-macrophage phagocytic activation (oxidative burst). Macrophage cytokine expression in response to control, base, and modified materials was determined by ELISAs for TNF-alpha, IL-1 beta, IL-6, and IL-8. Tissue culture poly(styrene) (TCPS)-mediated the greatest number of adherent monocyte/macrophage cells relative to PEG-g-PA materials. Both YRGDS and YEILDV peptides, whether directly or indirectly (via StarPEO) conjugated to PEG-g-PA, increased adhesion versus controls. Fab' fragments of all four antibodies also promoted enhanced adhesion versus controls. Fab'StarPEO materials presented two orders of magnitude fewer ligands per surface unit area than peptide star materials (10(8) vs. 10(10)), but were able to adhere similar numbers of cells. For surfaces presenting Fab'(VLA-4) or YEILDV, both of which may both bind to a cell's VLA-4 receptor, the Star:VLA4 surface showed a greater number of adherent monocyte/macrophages. This result suggests that the Fab' had a higher affinity to the cell receptor than a corresponding minimal peptide binding sequence. All materials exhibited low oxidative burst (luminescence counts per minute, LCPM) per cell DNA without the addition of exogenous stimuli (LCPM/DNA < 100). Directly conjugated peptide materials, poly(propylene) (PP), and TCPS showed the lowest levels of LCPM/DNA without the addition of exogenous stimulus (LCPM/DNA < 20). There was no correlation between LCPM/DNA ratios, with and without added LPS stimulus, versus the individual substrates. Monocyte/macrophages adherent to TCPS substrata showed the overall highest stimulatory potential in cytokine expression response to exogenous LPS, followed by PP>> PEG-g-PA>> StarPEO. Cells adherent to peptide-modified materials and Fab'-modified materials were overall less stimulated. The method of presenting the peptides (i.e., directly or via Star PEO) influenced the level of cytokine secreted by the adherent macrophage.

Encapsulating Au nanoparticles within a shell of photo-cross-linked block copolymer surfactant dramatically improves the physical and chemical stability of the nanoparticles, particularly when they are applied as bioconjugates. Photo-cross-linkable block copolymer amphiphiles [polystyrene-co-poly(4-vinyl benzophenone)]-block-poly(acrylic acid) [(PS-co-PVBP)-b-PAA] and [poly(styrene)-co-poly(4-vinyl benzophenone)]-block-poly(ethylene oxide) [(PS-co-PVBP)-b-PEO] were assembled around Au nanoparticles ranging from 12 to 108 nm in diameter. UV irradiation cross-linked the PVBP groups on the polymer to yield particles that withstood extremes of temperature, ionic strength, and chemical etching. Streptavidin was attached to [PS-co-PVBP]-b-PAA-coated particles using the same noncovalent and covalent conjugation protocols used to bind biomolecules to divinylbenzene-cross-linked PS microspheres. We expect that these particles will be useful as plasmonic, highly light-scattering and light-absorbing analogs to fluorescently labeled PS nanospheres.

Utilizing a first-principles-based coarse-grained implicit solvent model, we have investigated the self-association of C(60) fullerenes that have been symmetrically modified with six grafted poly(ethylene oxide) (PEO) chains in aqueous solution. Despite the highly symmetric nature of the pair interactions between PEO-grafted fullerenes, their supramolecular assemblies are highly anisotropic and resemble the linear clusters formed in Stockmayer fluids. The dipole-like interaction between these symmetrically modified fullerenes results from the shielding of the C(60) fullerenes by PEO, favoring the addition of more PEO-grafted fullerenes to the linear clusters at the relatively unprotected ends. At low nanoparticle concentrations, self-association is dominated by the formation of stable dimers and trimers resulting from fullerene-fullerene contact and favorable PEO-fullerene interactions. With increasing nanoparticle concentration, larger clusters become increasingly probable. The molecular weight of the PEO tethers can be treated as a temperature-like analogue, with a reduction in average cluster size with increasing chain length due to increased steric repulsion, which is qualitatively similar to effects observed in Stockmayer fluids with increasing temperature. The role of PEO in supramolecular self-organization in PEO-modified C(60) fullerene/water solutions is complex, contributing not only to steric stabilization but also to favorable energetic interactions, nanoparticle shielding, and depletion-driven aggregation.

Novel water-soluble amphiphilic triblock copolymers poly(glycerol monomethacrylate)-b-poly(propylene oxide)-b-poly(glycerol monomethacrylate) (PGMA-b-PPO-b-PGMA) were synthesized because of their expected enhanced ability to interact with biological membranes compared to the well-known poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) (PEO-b-PPO-b-PEO) block copolymers. Their bulkier hydrophilic PGMA blocks might induce a disturbance in the packing of liquid-crystalline lipid bilayers in addition to the effect caused by the hydrophobic PPO block alone. To gain a better insight into the polymer-membrane interactions at the molecular level, the adsorption kinetics and concomitant interactions of (PGMA14)(2-)PPO(34) with model membranes of dipalmitoylphosphatidylcholine (DPPC) and dimyristoylphosphatidylcholine (DMPC) were monitored using infrared reflection absorption spectroscopy (IRRAS) coupled with Brewster angle microscopy (BAM) and surface pressure (pi) measurements. The maximum penetration surface pressure of ca. 39 mN/m suggests that (PGMA14)(2-)PPO(34) is able to insert into lipid monolayers even above the so-called monolayer-bilayer equivalent pressure of 30-35 mN/m. Copolymer adsorption to a liquid-expanded DPPC-d62 monolayer proceeds in a two-step mechanism: (i) initially only the more hydrophobic PPO middle block penetrates the lipid monolayer; (ii) following the liquid-expanded-liquid-condensed (LE-LC) phase transition, the bulky PGMA hydrophilic blocks are dragged into the headgroup region as the PPO block inserts further into the fatty acid region. The adsorption kinetics is considerably faster for DMPC-d54 monolayers due to their higher fluidity. Copolymer adsorption to an LC-DPPC-d62 monolayer leads to a change in the monolayer packing by forcing the lipid alkyl chains into a more vertical orientation, their tilt angle with respect to the surface normal being reduced from initially 30 degrees +/- 3 degrees to 18 degrees +/- 3 degrees. BAM images rule out macroscopic phase separation and show that coalescence of DPPC-d62 LC domains takes place at relatively low surface pressures of pi>> or = 23 mN/m, suggesting that (PGMA14)(2-)PPO (34) partitions into both LE as well as LC domains.

Microbial adhesion to surfaces often occurs despite high wall shear rates acting on the adhering microorganisms. In this paper, we compare the wall shear rates needed to prevent microbial adhesion to bare glass and poly(ethylene oxide) (PEO)-brush coated glass in a parallel plate flow chamber. Initial microbial deposition rates were determined for different wall shear rates between 4 and 1600 s(-1) on the top and bottom plates of the flow chamber. Deposition efficiencies alpha(SL), based on the Smoluchowski-Levich approach, for Pseudomonas aeruginosa D1, Escherichia coli O2K2 and Candida tropicalis GB 9/9 decreased with increasing wall shear rates and were lower for PEO-brush coated glass than for bare glass. Characteristic shear rates preventing adhesion to the bottom plate were around 10 and 1.0 s(-1) for the bacteria on glass and the PEO-brush and 36 and 3.4s(-1) for the yeast strain on glass and the PEO-brush, respectively. This demonstrates that the adhesive forces between microorganisms and a PEO-brush are comparatively weak, although some strains may have the ability to adhere to a PEO-brush under low shear conditions. Microbial deposition efficiencies alpha(SL) were much larger, however, than unity for bottom plate deposition, but could be reduced to realistic values by averaging the deposition rates found for the top (negative contribution of sedimentation) and bottom (positive contribution of sedimentation) plates.

Patterning technologically important semiconductor interfaces with nanoscale metal films is important for applications such as metallic interconnects and sensing applications. Self-assembling block copolymer templates are utilized to pattern an aqueous metal reduction reaction, galvanic displacement, on silicon surfaces. Utilization of a triblock copolymer monolayer film, polystyrene-block-poly(2-vinylpyridine)-block-poly(ethylene oxide) (PS-b-P2VP-b-PEO), with two blocks capable of selective transport of different metal complexes to the surface (PEO and P2VP), allows for chemical discrimination and nanoscale patterning. Different regions of the self-assembled structure discriminate between metal complexes at the silicon surface, at which time they undergo the spontaneous reaction at the interface. Gold deposition from gold(III) compounds such as HAuCl4(aq) in the presence of hydrofluoric acid mirrors the parent block copolymer core structure, whereas silver deposition from Ag(I) salts such as AgNO3(aq) does the opposite, localizing exclusively under the corona. By carrying out gold deposition first and silver second, sub-100-nm gold features surrounded by silver films can be produced. The chemical selectivity was extended to other metals, including copper, palladium, and platinum. The interfaces were characterized by a variety of methods, including scanning electron microscopy, scanning Auger microscopy, X-ray photoelectron spectroscopy, and atomic force microscopy.

The function of a stimuli-responsive polymer was studied for the utilization of protein unfolding and refolding in protein separation using aqueous two-phase systems (ATPS). Poly(ethylene glycol) (PEG) bound to a thermo-reactive hydrophobic head (poly(propylene oxide)-phenyl group (PPO-Ph group)) was used as the functional ligand to modify the PEG phase of the aqueous two-phase systems. Firstly, refolding of carbonic anhydrase from bovine (CAB) was examined in the presence of PPO-Ph-PEG at various temperatures. The refolding yield of CAB was strongly enhanced and aggregate formation was suppressed by addition of PPO-Ph-PEG at a specific temperature (50-55 degrees C). The change in the local hydrophobicity of CAB and PPO-Ph-PEG was characterized using the aqueous two-phase partitioning method and a hydrophobic fluorescent probe. The local hydrophobicity of CAB was maximized at 60 degrees C. The local hydrophobicity of PPO-Ph-PEO was also found to be increased above 45 degrees C. A simple model for CAB refolding, which includes (i) PPO-Ph-PEG complex formation and CAB in the intermediate state and (ii) refolding and release of native CAB from the PPO-Ph-PEG surface, is suggested based on the evaluated surface hydrophobicity.