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PubMed1998

Catalytic significance of the specificity of divalent cations as KS* and kcat* cofactors for secreted phospholipase A2.

Calcium is required for the substrate binding and for the chemical step of the interfacial catalytic turnover cycle of pancreatic phospholipase A2 (PLA2), but not for the binding of the enzyme to the interface. The role of calcium and other divalent cations (C) is analyzed for the effect on the substrate binding and kcat* for the chemical step. The cofactor role of 3d-cations(II) (C) for the hydrolysis of dimyristoylphosphatidylmethanol (DMPM) vesicles is characterized as an equilibrium dissociation constant for the interfacial binary (E*C) and ternary (E*CL) complexes of PLA2 and substrate mimics (L). Of the cations(II) that promote the binding of a mimic to the enzyme at the interface (E*), only a subgroup supports the chemical step. For example, Cd, Zn, and Cu form ternary E*CL complexes with kcat* of <1 s-1, compared to the rate of >100 s-1 with Ca, Fe, Mn, Co, and Ni. Oxygen exchange from H218O to the products of hydrolysis of DMPM incorporates one 18O in myristate. Incorporation of the first and second 18O occurs during the incubation of both the products of hydrolysis in H218O with PLA2 and Ca, but not with Zn. The cation-dependent changes in the UV difference spectrum, associated with the formation of E*C and E*CL, suggest that the changes are mainly due to catalytic His-48, and possibly Tyr-52 and Tyr-73, and are different with Ca as opposed to Zn. These results and simulations suggest considerable plasticity in the calcium binding and catalytic site environment. It is proposed that the higher ground state stability of the E*CS complex with the inhibitory cations increases the effective activation energy. For the chemical step, calcium coordinated with a nucleophilic water and the ester carbonyl oxygen facilitates the near-attack geometry in the E*CaS, and the His-48.Asp-99 pair acts as a proton acceptor. As a prelude to establishing the catalytic mechanism, factors controlling the energetically demanding transition state are also discussed.

Animals↗
PubMed2004

Structural and dynamical characterization of Hele-Shaw viscous fingering.

Viscous fingering occurs in the interfacial zone between two fluids confined between two plates with a narrow gap (Hele-Shaw geometry) when a highly viscous fluid is displaced by a fluid with relatively low viscosity. Using a mesoscopic approach--the lattice Boltzmann method--we investigate the dynamics of spatially extended Hele-Shaw flow under conditions corresponding to various experimental systems by tuning the 'surface tension' and the reactivity between the two fluids. We discuss the onset of the fingering instability (dispersion relation), analyse the structural properties (characterization of the interface) and the dynamical properties (growth of the mixing zone) of the Hele-Shaw systems, and show the effect of reactive processes on the structure of the interfacial zone.

Journal Article↗
PubMed2006

Structure and dynamics of the aqueous liquid-vapor interface: a comprehensive particle-based simulation study.

This research addresses a comprehensive particle-based simulation study of the structural, dynamic, and electronic properties of the liquid-vapor interface of water utilizing both ab initio (based on density functional theory) and empirical (fixed charge and polarizable) models. Numerous properties such as interfacial width, hydrogen bond populations, dipole moments, and correlation times will be characterized with identical schemes to draw useful conclusions on the strengths and weakness of the proposed models for interfacial water. Our findings indicate that all models considered in this study yield similar results for the radial distribution functions, hydrogen bond populations, and orientational relaxation times. Significant differences in the models appear when examining both the dipole moments and surface relaxation near the aqueous liquid-vapor interface. Here, the ab initio interaction potential predicts a significant decrease in the molecular dipole moment and expansion in the oxygen-oxygen distance as one approaches the interface in accordance with recent experiments. All classical polarizable interaction potentials show a less dramatic drop in the molecular dipole moment, and all empirical interaction potentials studied yield an oxygen-oxygen contraction as the interface is approached.

Computer Simulation↗
PubMed1995

Characterization of the chemical bonding in inner layers of composite materials.

The efficiency of near edge structure investigations in electron energy loss spectroscopy (EELS) is discussed for characterizing the chemical bonding of elements present in the interfacial zone in fibre/matrix composites at nanometre resolution. Two different examples of corresponding analyses are given for a SiC-fibre reinforced borosilicate glass. In particular, the chemical bonding between silicon and carbon or oxygen (e.g. SiC, SiO(2) and SiO(x)C(y)), respectively, is characterized. The results have been attained in a fingerprint manner by comparing the fine structure measured from a material of unknown stoichiometry to that of a standard specimen. In addition, a possibility is demonstrated to image the chemical bonding by energy-filtered microscopy using energy loss near edge structures (ELNES).

Journal Article↗
PubMedDate not supplied

Effect of primer solvent and curing mode on dentin shear bond strength and interface morphology.

UNLABELLED: There is a need to study the main and interactive bonding effects of differences in solvent and curing mode used for adhesive monomers in dentin bonding systems. OBJECTIVE: Two solvents (acetone and ethanol) and curing methods (light cure, dual cure) were evaluated on their effects on bond strength and interfacial morphology. METHOD AND MATERIALS: The adhesives studied were based on two monomers, pyromellitate of glyceryl dimethacrylate (PMGDM) and 2-hydroxy ethyl methacrylate (HEMA). Four groups of eight teeth each were cut to expose planar dentin sections and treated with (a) light-cure system with acetone as solvent (LCA group); (b) light-cure system with ethanol as solvent (LCE group); (c) dual-cure system with acetone as solvent (DCA group); and (d) dual-cure system with ethanol as solvent (DCE group). The treated sections were tested for shear bond strength to composite discs and interfacial morphology. RESULTS: The mean (standard deviation) of shear bond strength values (MPa) for the different groups were: LCA: 11.8 (2.3); LCE: 12.7 (2.7); DCA: 24.9 (9.3); and DCE: 21.6 (9.6). All bonded sections were characterized by a similar hybrid layer, resin tags, and overall interfacial morphology. CONCLUSION: There was a significant difference in shear bond strength as a function of cure mode, but not of solvent. The mean bond strength was higher for dual-cure systems studied. Oxygen inhibition effects may account for the difference between light-cure and dual-cure types.

Acetone↗
PubMed2002

Molecular rulers: new families of molecules for measuring interfacial widths.

Homologous series of solvatochromic neutral alcohols and ionic sulfates are synthesized and characterized. Each surfactant series consists of hydrophobic, p-nitroanisole-based chromophores attached to polar or ionic headgroups by n-alkyl spacers. UV absorption measurements show that the optical properties of surfactant chromophores closely track those of the parent chromophore. Interfacial tension measurements are used to calculate surface excess concentrations of ionic surfactants adsorbed to an aqueous-cyclohexane interface. With a hydrophobic chromophore, a hydrophilic headgroup, and a variable-length, alkyl spacer, these surfactants have the potential to function as molecular rulers: probes of molecular-scale variation in solvation forces across condensed-phase interfaces. Changing the separation between the hydrophobic, solvatochromic probe and the hydrophilic headgroup should enable different members of a homologous series to span different interfacial widths, thus exposing the chromophore to different chemical environments. This idea is explored by using surface-specific, nonlinear optical spectroscopy. Resonant second harmonic spectra of p-nitroanisole and the surfactant product 4a adsorbed to an aqueous-cyclohexane interface show the surfactant spectrum blue-shifted 9 nm relative to the spectrum of adsorbed p-nitroanisole. On the basis of chromophore solvatochromism, these results are consistent with a less polar environment surrounding the surfactant chromophore. Significant differences in interfacial solvation resulting from a approximately 5 A separation between the surfactant headgroup and chromophore support recently proposed models of molecularly sharp, microscopically flat aqueous-alkane interfaces.

Journal Article↗
PubMed2006

A new technique for assessing hybrid layer interfacial micromorphology and integrity: two-photon laser microscopy.

PURPOSE: This study describes a two-photon laser fluorescence microscopy technique developed to evaluate the interfacial micromorphology of the hybrid layer in bonded restorations. MATERIALS AND METHODS: Micropermeability of the hybrid layer was characterized by means of simultaneously contrasting a dye-containing adhesive with a differently colored dye placed into the pulp chamber and allowed to diffuse toward the different-colored hybrid layer. A fluorescent red dye (rhodamine B) was incorporated into a commercial dentin bonding agent. Class I preparations (margins in enamel) were made on extracted human third molars. The teeth were restored using conventional methods: bonding agent, composite, finishing, and polishing. An aqueous solution of a yellow/green dye (fluorescein) was then placed into the pulp chamber for 3 h, allowing time to diffuse toward the different-colored bonded interface. The teeth were then embedded, sectioned, and microscopically analyzed using two-photon laser microscopy at 40X magnification. RESULTS: Subsurface fluorescent imaging using this technique enabled interfacial micromorphology to be characterized at submicrometer resolution and provided high-contrast images. The quality of surrounding structures and potential presence of gaps were also precisely assessed. CONCLUSION: Two-photon laser microscopy provided high quality, high-resolution images of the bonded interface and surrounding areas, allowing accurate qualitative and quantitative analysis of the structure and integrity of the hybrid layer.

Bisphenol A-Glycidyl Methacrylate↗
PubMed1997

Titration calorimetry of anesthetic-protein interaction: negative enthalpy of binding and anesthetic potency.

Anesthetic potency increases at lower temperatures. In contrast, the transfer enthalpy of volatile anesthetics from water to macromolecules is usually positive. The transfer decreases at lower temperature. It was proposed that a few selective proteins bind volatile anesthetics with negative delta H, and these proteins are involved in signal transduction. There has been no report on direct estimation of binding delta H of anesthetics to proteins. This study used isothermal titration calorimetry to analyze chloroform binding to bovine serum albumin. The calorimetrically measured delta H cal was -10.37 kJ.mol-1. Thus the negative delta H of anesthetic binding is not limited to signal transduction proteins. The binding was saturable following Fermi-Dirac statistics and is characterized by the Langmuir adsorption isotherms, which is interfacial. The high-affinity association constant, K, was 2150 +/- 132 M-1 (KD = 0.47 mM) with the maximum binding number, Bmax = 3.7 +/- 0.2. The low-affinity K was 189 +/- 3.8 M-1 (KD = 5.29 mM), with a Bmax of 13.2 +/- 0.3. Anesthetic potency is a function of the activity of anesthetic molecules, not the concentration. Because the sign of delta H determines the temperature dependence of distribution of anesthetic molecules, it is irrelevant to the temperature dependence of anesthetic potency.

Anesthetics↗
PubMed2002

Sustained ex vivo skin antiseptic activity of chlorhexidine in poly(epsilon-caprolactone) nanocapsule encapsulated form and as a digluconate.

In this work, the sustained bactericidal activity of chlorhexidine base loaded poly(epsilon-caprolactone), PCL, nanocapsules against Staphylococcus epidermidis inoculated onto porcine ear skin was investigated. Drug loaded nanocapsules were prepared by the interfacial polymer deposition following solvent displacement method, then characterized by photon correlation spectroscopy, electrophoretic measurements, transmission and scanning electron microscopy. Antimicrobial activity of these colloidal carriers was evaluated (i) in vitro against eight strains of bacteria, and (ii) ex vivo against Staphylococcus epidermidis inoculated for 12 h onto porcine ear skin surface treated for 3 min either with 0.6% chlorhexidine base loaded or unloaded nanocapsules suspended in hydrogel, or 1% chlorhexidine digluconate aqueous solution. Chlorhexidine absorption into the stratum corneum (SC) was evaluated by the tape-stripping method. The results showed that chlorhexidine nanocapsules in aqueous suspension having a 200-300 nm size and a positive charge exhibited similar minimum inhibitory concentrations against several bacteria with chlorhexidine digluconate aqueous solution. Ex vivo, there was a significant reduction in the number of colony forming units (CFUs) from 3-min treated skin with chlorhexidine nanocapsule suspension (5 to <1 log(10)) compared to chlorhexidine digluconate solution (5 to 2.02 log(10)) after a 8-h artificial contamination. After a 12-h artificial contamination, both formulations failed to achieve a 5 log(10) reduction. Furthermore, from a 3-min treatment with an identical applied dose and a subsequent 12-h artificial contamination, a residual chlorhexidine concentration in the SC was found to be three-fold higher with chlorhexidine nanocapsule suspension than with chlorhexidine digluconate solution. Interestingly, nanocapsules were shown in porcine skin follicles. Consequently, a topical application of chlorhexidine base-loaded positively charged nanocapsules in an aqueous gel achieved a sustained release of bactericide against Staphylococcus epidermidis for at least 8 h. Enhancement of drug delivery by mediating a more direct and prolonged contact between the carrier and (i) bacteria, (ii) skin surface, and (iii) skin follicles was assumed.

Animals↗
PubMed2002

In vitro inhibition of bovine herpes virus 1 reproduction with native and microencapsulated proteinase inhibitor aprotinin.

This study evaluated the antiviral effect of various dosage forms of proteinase inhibitor-aprotinin as a potential remedy for prophylactics and therapy of infectious bovine rhinotracheitis. Formulations of the inhibitor were tested for their influence on bovine herpes virus reproduction in cell cultures. Starch/bovine serum albumin microcapsules with aprotinin were prepared using interfacial cross-linking with terephthaloyl chloride and characterized for their morphology, size and release of the inhibitor. Two types of these microcapsules-impregnated and loaded with the inhibitor-were used in virus infectious studies. Native aprotinin possessed palpable dose-dependent antiviral effect inhibiting the virus reproduction up to 4.0 lg (10000-fold) and delaying the cytopathic effect up to 96 h in the concentration 800-3300 TIU/ml. The bioadhesive, biodegradable aprotinin-loaded microcapsules were the most effective antiviral drug as this formulation allowed to decrease virus infectious titer up to 4.0 lg and a delay in the cytopathic effect of up to 144 h in lesser doses of inhibitor compared with the native form. In comparison the antiviral effect of microcapsules impregnated with aprotinin was not so appreciable. It was interesting to note that the results of the experiments on diverse cultures were very similar. This was because the drugs influenced the fundamental processes of virus replication cycle.

Animals↗
PubMedDate not supplied

Structure of hollow polystyrene microspheres: an SEM study.

Hollow polystyrene microspheres have been prepared by a modified interfacial polymer deposition technique. The hollow microspheres were characterized using scanning electron microscopy. The effects of polymer concentration and the quantity of the internal phase material (methanol) used on the microsphere size and their wall thickness were studied. Increase in polymer concentration causes increase in wall thickness while increasing internal phase material increases microsphere size and deforms them from a spherical shape.

Microscopy, Electron, Scanning↗
PubMed2006

Mechanical properties of interfacial films formed by lysozyme self-assembly at the air-water interface.

We present the first characterization of the mechanical properties of lysozyme films formed by self-assembly at the air-water interface using the Cambridge interfacial tensiometer (CIT), an apparatus capable of subjecting protein films to a much higher level of extensional strain than traditional dilatational techniques. CIT analysis, which is insensitive to surface pressure, provides a direct measure of the extensional stress-strain behavior of an interfacial film without the need to assume a mechanical model (e.g., viscoelastic), and without requiring difficult-to-test assumptions regarding low-strain material linearity. This testing method has revealed that the bulk solution pH from which assembly of an interfacial lysozyme film occurs influences the mechanical properties of the film more significantly than is suggested by the observed differences in elastic moduli or surface pressure. We have also identified a previously undescribed pH dependency in the effect of solution ionic strength on the mechanical strength of the lysozyme films formed at the air-water interface. Increasing solution ionic strength was found to increase lysozyme film strength when assembly occurred at pH 7, but it caused a decrease in film strength at pH 11, close to the pI of lysozyme. This result is discussed in terms of the significant contribution made to protein film strength by both electrostatic interactions and the hydrophobic effect. Washout experiments to remove protein from the bulk phase have shown that a small percentage of the interfacially adsorbed lysozyme molecules are reversibly adsorbed. Finally, the washout tests have probed the role played by additional adsorption to the fresh interface formed by the application of a large strain to the lysozyme film and have suggested the movement of reversibly bound lysozyme molecules from a subinterfacial layer to the interface.

Air↗
PubMed2006

The interfacial binding surface of phospholipase A2s.

For membrane-associated enzymes, which access substrate from either a monolayer or bilayer of the aggregate substrate, the partitioning from the aqueous phase to this phospholipid interface is critical for catalysis. Despite a large and expanding body of knowledge regarding interfacial enzymes, the biophysical steps involved in interfacial recognition and adsorption remain relatively poorly understood. The surface of the enzyme that contacts the phospholipid surface is referred to as its interfacial binding surface, or more simply, its i-face. The interaction of a protein's i-face with the aggregate substrate may simply control access to substrate. However, it can be more complex, and this interaction often serves to allosterically activate the enzyme on this surface. First we briefly review what is currently known about i-face structure and function for a prototypical interfacial enzyme, the secreted Phospholipase A2 (PLA2). Then we develop, characterize, compare, and discuss models of the PLA2 i-face across a subset of five homologous PLA2 family members, groups IA, IB, IIA, V, and X. A homology model of human group-V is included in this comparison, suggesting that a similar approach could be used to explore interfacial function of any of the PLA2 family members. Despite moderate sequence identity, structural homology and sequence similarity are well conserved. We find that the residues predicted to be interfacial, while conserved structurally, are not highly conserved in sequence. Implications for this divergence on interfacial selectivity are discussed.

Humans↗
PubMed2000

Interfacial properties of barium sulfate suspensions. Implications in their stability.

A surface characterization of barium sulfate particles in aqueous suspensions was carried out in this work. With the aim of predicting the stability conditions of these widely used suspensions, the electrical surface properties of the particles were first studied by electrophoretic mobility determinations. It was found that both H(+) and OH(-) ions can be considered as potential-determining ions for the barium sulfate/water interface. The same conclusion was reached concerning the lattice ions, Ba(2+) (mainly) and SO(4)(2-). It was also found that increasing the concentration of sodium chloride in the dispersion medium can even change the sign of the zeta (zeta) potential: it is suggested that this behavior is an indirect effect provoked by changes in the solubility of barium sulfate with the ionic strength. To compute the van der Waals (LW) attraction between the particles as well as the acid-base contribution to the total energy of interaction, a thermodynamic characterization of the interface was also carried out by measuring the rate of penetration of selected liquids through plugs of the particles. It was found that barium sulfate particles are essentially monopolar in nature; that is, they show electron-donor character as demonstrated by the essentially zero value of the electron-acceptor component of their surface free energy. Pretreatment of the particles with 10(-2) M solutions of BaCl2 and CaCl2 significantly reduced the electron-donor component, whereas both NaCl and Na(2)SO(4) provoked the opposite effect. This result is explained in terms of the acid-base character of the ions added. These data were used to calculate the interaction energy between the particles. The effect of electrolyte concentration on the stability of the suspensions was analyzed on the basis of the dependence with distance of the interaction energy between the particles. Our results suggest that more stable suspensions of barium sulfate are predicted if moderate amounts of SO(4)(2-) ions are added to the dispersion medium.

Barium Sulfate↗
PubMed2003

Bonding mechanism and ultrastructural interfacial analysis of a single-step adhesive to dentin.

OBJECTIVES: to characterize the interaction of 4-acryloxyethyltrimellitic acid (4-AET), a key ingredient in a commercial all-in-one adhesive (Reactmer Bond) with dentin apatite, to measure the solubility of 4-AET calcium (4-AETCa), and to examine the ultrastructure of the resin-dentin interface bonded by this adhesive. METHODS: solubility of synthesized 4-AETCa in several solutions was measured. Flattened bovine dentin blocks were treated with 0.1wt% NaOCl for 10 min, or 5.0wt% NaOCl for 10 min, or 30 min. Fourier transform infrared analysis using attenuated total reflection (FT-IR/ATR) was performed on these four substrates before and after treatment with a 4-AET mixture (4-AET/HEMA: 40/60wt%). Undemineralized, unstained sections of human sound dentin that were bonded with the same comonomers were examined by transmission electron microscopy (TEM). RESULTS: the respective solubilities of the 4-AETCa in water, 2-hydroxyethyl methacrylate (HEMA; 100wt%) and HEMA/water (70/30wt%) were 0.023, 0.003 and 0.014 mol/l after 2 min. The IR analysis indicated that Ca-carboxylate (new bands: nuCz.dbnd6;O:1580 and 1413 cm(-1)) was formed on all dentin specimens following application of 4-AET, with or without NaOCl pretreatment. TEM showed a 1 micro m thick hybrid layer with some remnant apatite crystallites that were encapsulated by a layer of less electron-dense material. SIGNIFICANCE: the results suggest that ionized carboxyl groups in 4-AET may interact with Ca(2+) from apatite crystallites within the partially demineralized hybrid layer to form an insoluble calcium salt (4-AETCa) that may aid in bonding this resin system to dentin.

Animals↗
PubMed2005

Adsorption of bituminous components at oil/water interfaces investigated by quartz crystal microbalance: implications to the stability of water-in-oil emulsions.

Silica-gel-coated QCM crystals oscillating in a thickness shear mode are used to measure adsorption of bituminous components in water-saturated heptol (1/1 vol ratio of a heptane/toluene mixture) at the oil/water interface. In addition to the viscoelasticity of the adsorbed film, the effects of the bulk liquid density and viscosity as well as the liquid trapped in interfacial cavities are taken into account for the calculation of adsorbed mass. Asphaltenes in heptol adsorb continuously at the oil/water interface, while resins (the surface-active species in maltenes) show adsorption saturation in the same solvent. For Athabasca bitumen in heptol, two adsorption regimes are observed depending on concentration. At low concentrations, a slow, non-steady-state, and irreversible adsorption takes place. At high concentrations, a steady-state adsorption with limited reversibility results in a quick adsorption saturation. The threshold concentration between these adsorption regimes is 1.5 wt % and 8 wt % for oil/water and oil/gold interfaces, respectively. The threshold concentration, the total adsorbed amount, and the flux of non-steady-state adsorption depend on the resin-to-asphaltene ratio. The threshold concentration is related to the earlier reported critical bitumen concentration characterizing the rigid-to-flexible transition of the interfacial film. We propose a new mechanism based on the change of the effective resin-to-asphaltene ratio with dilution to explain both the adsorption behavior and emulsion stability.

Journal Article↗
PubMed2006

Colloidal interactions between asphaltene surfaces in aqueous solutions.

Asphaltene at oil/water interfaces plays a dominant role in the recovery of crude oil. In this study, asphaltene monolayer films were deposited on hydrophobic silicon wafers and silica spheres from oil-water interfaces using a Langmuir interfacial trough. The morphology of the deposited asphaltene films was characterized with an atomic force microscope (AFM). The colloidal forces between the prepared asphaltene films in aqueous solutions were measured with AFM to shed light on the stabilization of water or oil droplets coated with asphaltene films. Factors such as solution pH, KCl concentration, calcium addition, and temperature all showed a strong impact on colloidal forces between the prepared asphaltene films. The findings provided a better understanding of asphaltene interfacial films at an oil/water interface in stabilizing bitumen-in-water and water-in-bitumen emulsions.

Journal Article↗
PubMed2000

Enhancement of mass transfer using colloidal liquid aphrons: measurement of mass transfer coefficients in liquid-liquid extraction.

Interphase mass transfer of a sparingly soluble solute is often the rate-limiting step in multiphase biocatalytic processes. Colloidal liquid aphrons (CLA) provide very large interfacial areas, and thus could enhance mass transfer in such processes. The aim of this study was to characterize mass transfer properties of CLA dispersions during transfer of heptanoic acid from water to limonene. The interfacial area per unit volume (a), film mass transfer coefficient (K(L)), and volumetric mass transfer coefficient (K(L)a) values were determined in a stirred-tank reactor. These results were used, along with a literature correlation, to estimate the mass transfer coefficient of the surfactant-stabilized shell surrounding the CLA. The very large increase in a provided by the CLA was only partially offset by a slight increase in the mass transfer resistance of the shell. As a result, the range of K(L)a values obtained using CLA was about an order of magnitude greater than that obtained using a conventional dispersion. The concentration of the aqueous-phase surfactant used to form the CLA strongly affected the Sauter mean diameter of the CLA; however, the concentration of the nonpolar-phase surfactant had little effect. These results suggest that CLA have considerable potential for multiphase biocatalytic applications.

Biotechnology↗