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Amino acids bracketing the predicted transmembrane domains of membrane proteins.

The cell membrane is a complex mixture of several classes of biomolecules but amino acids and lipids are the main constituents. For this reason we are establishing a data base of transmembrane proteins with the intent of using the data base to identify interfacial peptide sequences useful for studying protein-lipid interactions at membrane interfaces. Our present intention is to characterize transmembrane peptides and amino acids found near the membrane interface. A data base containing only signal peptides is available (G. von Heijne, Prot. Seq. Data Anal. 1:41-42, 1987).

Amino Acid Sequence↗

Fracture toughness of dentin/resin-composite adhesive interfaces.

The reliability and validity of tensile and shear bond strength determinations of dentin-bonded interfaces have been questioned. The fracture toughness value (KIC) reflects the ability of a material to resist crack initiation and unstable propagation. When applied to an adhesive interface, it should account for both interfacial bond strength and inherent defects at or near the interface, and should therefore be more appropriate for characterization of interface fracture resistance. This study introduced a fracture toughness test for the assessment of dentin/resin-composite bonded interfaces. The miniature short-rod specimen geometry was used for fracture toughness testing. Each specimen contained a tooth slice, sectioned from a bovine incisor, to form the bonded interface. The fracture toughness of an enamel-bonded interface was assessed in addition to the dentin-bonded interfaces. Tensile bond strength specimens were also prepared from the dentin surfaces of the cut bovine incisors. A minimum of ten specimens was fabricated for each group of materials tested. After the specimens were aged for 24 h in distilled water at 37 degrees C, the specimens were loaded to failure in an Instron universal testing machine. There were significant differences (p < 0.05) between the dental adhesives tested. Generally, both the fracture toughness and tensile bond strength measurements were highest for AllBond 2, intermediate for 3M MultiPurpose, and lowest for Scotchbond 2. Scanning electron microscopy of the fractured specimen halves confirmed that crack propagation occurred along the bond interface during the fracture toughness test. It was therefore concluded that the mini-short-rod fracture toughness test provided a valid method for characterization of the fracture resistance of the dentin-resin composite interface.

Analysis of Variance↗

Membrane perturbation induced by interfacially adsorbed peptides.

The structural and energetic characteristics of the interaction between interfacially adsorbed (partially inserted) alpha-helical, amphipathic peptides and the lipid bilayer substrate are studied using a molecular level theory of lipid chain packing in membranes. The peptides are modeled as "amphipathic cylinders" characterized by a well-defined polar angle. Assuming two-dimensional nematic order of the adsorbed peptides, the membrane perturbation free energy is evaluated using a cell-like model; the peptide axes are parallel to the membrane plane. The elastic and interfacial contributions to the perturbation free energy of the "peptide-dressed" membrane are evaluated as a function of: the peptide penetration depth into the bilayer's hydrophobic core, the membrane thickness, the polar angle, and the lipid/peptide ratio. The structural properties calculated include the shape and extent of the distorted (stretched and bent) lipid chains surrounding the adsorbed peptide, and their orientational (C-H) bond order parameter profiles. The changes in bond order parameters attendant upon peptide adsorption are in good agreement with magnetic resonance measurements. Also consistent with experiment, our model predicts that peptide adsorption results in membrane thinning. Our calculations reveal pronounced, membrane-mediated, attractive interactions between the adsorbed peptides, suggesting a possible mechanism for lateral aggregation of membrane-bound peptides. As a special case of interest, we have also investigated completely hydrophobic peptides, for which we find a strong energetic preference for the transmembrane (inserted) orientation over the horizontal (adsorbed) orientation.

Algorithms↗

A study of protein dynamics from anisotropy decays obtained by variable frequency phase-modulation fluorometry: internal motions of N-methylanthraniloyl melittin.

Internal motions of melittin and its lipid complexes were studied by anisotropy decays determined by frequency-domain fluorometry. A covalent anthraniloyl probe was attached, probably to lysine-21. The emission spectra indicate that the anthraniloyl moiety is exposed to solvent in both monomeric and tetrameric forms and is present at the lipid-water interfacial region in the lipid complexes. The fluorescence intensity decay of melittin in solution and its lipid complexes was characterized by three lifetimes. The lifetimes were near 1-2 ns, 6-7 ns and 10 ns. At increased temperatures there was an increase in the amplitude of the intermediate lifetime and a decrease in that of the longer lifetime. For all the melittin systems, at least three correlation times were required to fit the anisotropy data. Of the three correlation times, the shortest correlation time represents the local motions of the probe, while the longest represents global motions of the whole molecule. The intermediate correlation time probably represents the dynamics of domains/helices within the molecule. The melittin monomer is highly flexible, with greater than 90% of its anisotropy being lost by the local motions. Even though it is well organized (greater than 75% helical), the tetramer is still a highly flexible molecule, with 70% of its anisotropy being lost by the local motions. The internal motions of melittin decrease upon binding to lipids and are sensitive to the phase state of the lipid complexes.

Bee Venoms↗

Stepwise self-assembly of ordered supramolecular assemblies based on coordination chemistry.

Ultrathin multilayers based on transition metal complexes have been prepared by successive deposition and self-assembly. Dendrimer layers were deposited onto SiO2 wafers by alternately immersing the substrate into a solution of terpyridyl (tpy)-pendant poly(amido amine) (PAMAM) dendrimers (dend-n-tpy; n = 8, 16) dissolved in CH2Cl2, followed by the interfacial coordination reaction with cobalt (Co2+) from aqueous solution. The films derived from this simple assembly method have been characterized by electrochemical methods, synchrotron-based X-ray reflectivity (XRR), and X-ray fluorescence (XRF) recorded under grazing incidence. XRR analysis revealed a linear thickness dependence with an increase of 9.9 +/- 0.5 angstroms and 10.8 +/- 0.4 angstroms per growth cycle for both dend-8-tpy and dend-16-tpy, respectively, indicative of a layer-by-layer (LbL) growth of single dendrimer/Co2+ layers. XRF and electrochemical results showed that the amount of Co2+ increases linearly as more layers are deposited, and that the Co2+ concentration (mol/L) in dend-8-tpy/Co2+ films decays slowly as the number of growth cycles (l) increases. Moreover, a preliminary kinetics analysis indicated that the growth of a dendrimer layer in a deposition cycle is a self-limiting process.

Dendrimers↗

Effect of vertical temperature variation on the oscillatory wetting instability in a fluid Ga-Pb alloy.

Oscillatory wetting instabilities driven by capillary-gravitation forces have been explored very recently in the binary fluid Ga-Pb alloy [A. Turchanin, R. Tsekov and W. Freyland, J. Chem. Phys., 2004, 120, 11 171]. This system is characterized by a complete wetting transition at liquid-liquid coexistence. Due to its metallic nature the bulk and interfacial instabilities are strongly coupled via variation of the respective emissivities. In our previous work we have investigated these phenomena at different cooling cycles and at constant temperature inside the miscibility gap. In this study we present for the first time the observations of the oscillatory wetting instabilities also in heating cycles. The interfacial properties of a Ga0.95Pb0.05 alloy at conditions inside the miscibility gap have been investigated by following the second harmonic generation (SHG) intensity changes. Corresponding model calculations of the Pb-rich wetting film instabilities have been performed taking into account the effect of a temperature variation vertical to the bulk sample. The influence of this temperature variation on the occurrence of the oscillations is discussed.

Alloys↗

Interfaces between metal and arylamine molecular films as probed with the anode interfacial engineering approach in single-layer organic diodes.

The interfaces between metal and arylamine molecular films were studied by measurement of current-voltage characteristics in single-layer organic diodes with the aid of the anode interfacial engineering method. The diode turn-on voltage is shown to be very sensitive to the arylamine/metal interface and thus can serve as a means for characterizing such organic/metal interfaces. We show that the diode electrical performance depends on the arylamine substituent groups and chemical/electrical properties of the anode. The conduction mechanism of the diodes is shown to be injection limited, which can be well described with the Richardson-Schottky thermionic emission model. This work may provide some insight into the use of single-layer organic diodes and the interfacial engineering method to rapidly probe the organic/metal and even organic/organic interfaces.

Journal Article↗

Nominal shear or fracture mechanics in the assessment of composite-dentin adhesion?

This study addresses the anticipated problem of discriminating among high-performing dentin adhesives. The simplicity of the nominal shear bond test, despite being heavily criticized, has made it a routine procedure for the determination of bonding efficacy. A fracture mechanics approach has been suggested as a better assessment of bonding efficacy (Versluis et al., 1997). However, experimental complexity is a major limitation. It is hypothesized that a new, simplified interfacial fracture toughness test (Lin, 1994) will evaluate bonding agents differently if compared with the traditional shear bond test. Therefore, the objective of this study was to compare the performances of six dentin bonding agents subjected to the interfacial fracture toughness test (critical plane strain energy release rate) or to the nominal shear bond test (shear bond strength). Their performances were also characterized by scanning electron micrography of the fracture surfaces for evidence of dentin cohesive failure. Statistical analyses showed only marginal differences between these determinants of the two tests. However, when the analysis was applied only to the materials that had 100% frequency of dentin cohesive failure in shear testing, which also had high bonding efficacy, the difference in adhesive strengths between the two tests became significant. The reliability of the nominal shear test is questioned when dentin cohesive failure occurs, which usually is associated with high bonding efficacy. Since it is expected that bonding efficacy will increase further, the interfacial fracture toughness test is the preferred methodology to distinguish among high-performing dentin adhesives.

Adhesiveness↗

Selective labeling of pulmonary surfactant protein SP-C in organic solution.

Pulmonary surfactant protein SP-C has been isolated from porcine lungs and treated with dansyl isothiocyanate in chloroform:methanol 2:1 (v/v) solutions,under conditions optimized to introduce a single dansyl group covalently attached to the N-terminalamine group of the protein without loss of its native thioesther-linked palmitic chains. The resulting derivative Dans-SP-C conserves the secondary structure of native SP-C as well as the ability to promote interfacial adsorption of DPPC suspensions and to affect the thermotropic behavior of DPPC bilayers. This derivative can be used to characterize lipid-protein and protein-protein interactions of a native-like SP-C in lipid/protein complexes.

Animals↗

Assays of phospholipases on short-chain phospholipids.

Short-chain phospholipids are extremely useful compounds for analysis of interfacial and substrate requirements of water-soluble phospholipases. A wide variety of PCs are easily prepared and can be characterized in detail to the point where at a given PC concentration, the micelle size distribution is known, and an area per head group can be estimated. Since they can be presented to enzymes as monomers, micelles, or in bilayer structures, short-chain phospholipids are ideal for examining kinetic preferences of lipolytic enzymes. In the future detailed studies of short-chain phospholipids with different head groups should facilitate kinetics with phospholipases which show a preference for anionic substrates.

Animals↗

Surface properties of intraocular lens materials and their influence on in vitro cell adhesion.

An in vitro model to assess lens epithelial cell adhesion to a variety of intraocular lens materials was developed. Rabbit anterior lens capsules were isolated and cultured in serum-containing medium. Test surfaces included poly(methyl methacrylate), two new silicones (SLM-1/UV, SLM-2/UV), two hydrogels (HEMA, Lidofilcon A), and polytetrafluoroethylene (PTFE). Following the application and culturing of cells on the test surfaces, adherent cells were removed by trypsinization and counted at eight and 24 hours. The material surfaces were characterized by electron spectroscopy for chemical analysis and scanning electron microscopy. The captive bubble technique was also used to assess interfacial free energy. More cells adhered to PMMA than to the other materials tested (P less than .01). The two silicones, HEMA, and PTFE did not differ significantly from each other; Lidofilcon A had the lowest cell adhesion of all materials tested. Cell adhesion results were related to the interfacial free energy of each material. Materials of low (less than 5 ergs/cm2) or high (greater than 40 ergs/cm2) interfacial free energies had lower cell adhesion than materials of intermediate free energies (5 to 40 ergs/cm2) which exhibited the highest cell adhesion.

Animals↗

Furfural-cysteine model reaction in food grade nonionic oil/water microemulsions for selective flavor formation.

The thermal reaction between cysteine and furfural was investigated at 65 degrees C in five-component food grade oil/water (O/W) microemulsions of R-(+)-limonene/ethanol, EtOH/water/propylene glycol, PG/Tween 60 as apart of a systematic study on the generation of aroma compounds by utilizing structured W/O and O/W fluids. The furfural-cysteine reaction led to the formation of unique aroma compounds such as 2-furfurylthiol (FFT), 2-(2-furanyl)thiazolidine (main reaction product), 2-(2-furanyl)thiazoline, and N-(2-mercaptovinyl)-2-(2-furanyl)thiazolidine. These products were determined and characterized by GC-MS. Enhancement in flavor formation is termed "microemulsion catalysis". The chemical reaction occurs preferably at the interfacial film, and therefore a pseudophase model was assumed to explain the enhanced flavor formation. The product internal composition is dictated by process conditions such as temperature, time, pH, and mainly the nature of the interface. Increasing water/PG ratio leads to a dramatic increase in the initial reaction rate (V(0)). V(0) increased linearly as a function of the aqueous phase content, which could be due to the increase in the interfacial concentration of furfural. Microemulsions offer a new reaction medium to produce selective aroma compounds and to optimize their formation.

Cyclohexenes↗

Interfacial water on Cl- and H-terminated Si(111) surfaces from first-principles calculations.

The properties of interfacial water on Cl- and H-terminated Si(111) surfaces are investigated using a first-principles approach and characterized by means of energetic analysis combined with hydrogen-bond counting. The interaction of water with both substrates is found to be significantly weak, although bonding with the Cl-terminated Si(111) surface is relatively stronger because of the electrostatic contribution. According to a molecular picture for attributing the hydrophilic/hydrophobic character, both surfaces should be considered hydrophobic, at variance with the interpretation of recent ultrafast electron crystallography experiments, which seems instead to support a hydrophilic nature of the Cl-terminated Si(111) substrate.

Journal Article↗

Interactions of mouse Paneth cell alpha-defensins and alpha-defensin precursors with membranes. Prosegment inhibition of peptide association with biomimetic membranes.

The bactericidal activity of mouse alpha-defensins (cryptdins) requires proteolytic activation of inactive precursors by matrix metalloproteinase-7 (matrilysin, EC, MMP-7(a)). To investigate mechanisms of cryptdin-4 (Crp4) peptide interactions with membrane bilayers and to determine whether MMP-7-mediated proteolysis activates the membrane disruptive activity of Crp4, associations of Crp4 and melittin with biomimetic lipid/polydiacetylene chromatic vesicles were characterized. The peptides differ in their sensitivity to vesicle lipid composition and their depth of bilayer penetration. Crp4 undergoes strong interfacial binding onto lipid bilayers with disruption of the bilayer head group region, unlike melittin, which inserts more deeply into the hydrophobic core of the bilayer. Colorimetric and tryptophan fluorescence studies showed that Crp4 insertion is favored by negatively charged phospholipids and that zwitterionic and Escherichia coli phospholipids promote stronger interfacial binding; melittin-membrane interactions were independent of either variable. In contrast to the membrane disruptive activity of Crp4, pro-Crp4 did not perturb vesicular membranes, consistent with the lack of bactericidal activity of the precursor, and incubation of Crp4 with prosegment in trans blocked Crp4 and G1W-Crp4 membrane interactions at concentrations that inhibit Crp4 bactericidal activity. CD measurements showed that Crp4 has an expected beta-sheet structure that is not evident in the pro-Crp4 CD trace or when Crp4 is incubated with prosegment, indicating that the beta-sheet signal is attenuated by proregion interactions or possibly disrupted by the prosegment. Collectively, the results suggest that the prosegment inhibits Crp4 bactericidal activity by blocking peptide-mediated perturbation of target cell membranes, a constraint that is relieved when MMP-7 cleaves the prosegment.

Amino Acid Sequence↗

Polyalkyl cyanoacrylate nanocapsules.

In-situ polymerized methyl cyanoacrylate (MCA), ethyl cyanoacrylate (ECA), and butyl cyanoacrylate (BCA) were used to prepare nanocapsules of fluorescein or doxorubicin as markers by a w/o emulsion interfacial polymerization technique. Different concentrations of MCA were also used to show the effect of monomer concentration. The nanocapsules were characterized by electron microscopy, particle size analysis, holding capacity and in-vitro release of the marker substances. After selection of the polymerization solvent system, nearly spherical nanocapsules were obtained using each of the monomers. Most of the nanocapsules prepared were in the particle size range 500-1500 nm diameter. They were able to hold 55-74% of the marker initially present in aqueous solution. In-vitro dissolution studies showed that release of marker was retarded variably in an increasing order from nanocapsules containing MCA, ECA then BCA. Increasing the concentration of the monomer in the nanocapsules led to retardation of marker release.

Capsules↗

Characterization of the structures of poly(urea-urethane) microcapsules.

A series of poly(urea-urethane) microcapsules containing phthalate derivatives as a core material were prepared by an interfacial polymerization process in order to investigate the structural formation mechanism. Scanning electron microscopy (SEM) analysis for the cross sections of microcapsules revealed the systematic formations of porous structures followed by the formation of core/shell structures. Critical values of the core oil content for the formation of porous and core/shell structures were obtained from SEM results and the critical values were found to be proportional to the solubility parameters of core materials. Dynamic mechanical measurements indicated an amorphous structure of wall membrane and the glass transition temperature was found to decrease with increasing the core oil content suggesting a plasticizing effect. The surface amount of the core oils absorbed in the wall membrane was estimated using time of flight secondary ion spectroscopy analysis and found to increase with increasing the oil content before reaching constant. This tendency was interpreted in terms of the structural formation of the microcapsules. The results obtained in the present investigation were reasonably understood on the basis of swelling theory of wall membrane and the Flory-Huggins interaction parameters of the systems were discussed.

Algorithms↗

Interfaces and thin films as seen by bound electromagnetic waves.

This contribution summarizes the use of plasmon surface polaritons and guided optical waves for the characterization of interfaces and thin organic films. After a short introduction to the theoretical background of evanescent wave optics, examples are given that show how this interfacial "light" can be employed to monitor thin coatings at a solid/air or solid/liquid interface. Examples are given for a very sensitive thickness determination of samples ranging from self-assembled monolayers, to multilayer assemblies prepared by the Langmuir/Blodgett/Kuhn technique or by the alternate polyelectrolyte deposition. These are complemented by the demonstration of the potential of the technique to also monitor time-dependent processes in a kinetic mode. Here, we put an emphasis on the combination set-up of surface plasmon optics with electrochemical techniques, allowing for the on-line characterization of various surface functionalization strategies, e.g. for (bio-) sensor purposes.

Journal Article↗

Metal-matrix interface in reinforced glass ionomers.

OBJECTIVE: In reinforced materials, interfacial bonding is critical for efficient transfer of stress from the matrix to the reinforcement. The goal of this study was to characterize the physical and chemical nature of this interface in three metal-reinforced glass ionomers. Two of them were commercial, ESPE-Ketac Silver (KS) and GC-Miracle Mix (MM), and the third, EX, was experimental. METHODS: The techniques of scanning electron microscopy (SEM) and infrared spectroscopy (IR) were used to accomplish the stated goal. SEM analysis utilized polished sections of set cylindrical specimens of each material prepared from their respective powder and liquid components. The glass and the liquid in EX were the same as in MM. The reinforcing agent in EX, 50% by weight, was an Ag-base spherical alloy similar in composition to that in MM. For EX, the alloy was oxidized to promote its bonding to the matrix. The specimens for IR study were prepared as follows. The metallic powders of each material were mixed with the corresponding liquid in excess, stored at 37 degrees C for 1 h, washed with warm water (60 degrees C), filtered and dried. The untreated metallic powders served as controls for IR analysis. RESULTS: SEM revealed a distinct halo shaped internal reaction layer surrounding each alloy particle in EX. A similar layer was not seen in MM and KS. The alloy-matrix interface was continuous and gap-free in EX. In contrast, gaps separating matrix from respective reinforcements were conspicuous features in MM and KS. The IR spectrum of the liquid treated EX alloy powder showed absorbency bands characteristic of unreacted carboxyl groups and carboxylate salts. These bands were absent in the IR spectra of all other powders--treated and untreated. SIGNIFICANCE: The absence of interfacial bonding in MM and KS demonstrated in this study provides a reason why these two materials, in spite of metal addition, have not proved to be any stronger or more durable than their metal-free counterparts. A means of creating interfacial bonding presented here could be useful in the design of improved reinforced glass ionomer materials.

Aluminum↗