Biomedical subjects

Clayton J Radke

Publications and source records attributed to Clayton J Radke.

10 recordsLinked to original sources

PubMed2005

Steady-state diffusion of water through soft-contact-lens materials.

Water transport through soft contact lenses (SCL) is important for acceptable performance on the human eye. Chemical-potential gradient-driven diffusion rates of water through SCL materials are measured with an evaporation-cell technique. Water is evaporated from the bottom surface of a lens membrane by impinging air at controlled flow rate and humidity. The resulting weight loss of a water reservoir covering the top surface of the contact-lens material is recorded as a function of time. New results are reported for a conventional hydrogel material (SofLens One Day, hilafilcon A, water content at saturation w10 = 70 weight %) and a silicone hydrogel material (PureVision, balafilcon A, w10 = 36%), with and without surface oxygen plasma treatment. Also, previously reported data for a conventional 2-hydroxyethyl methacrylate (HEMA)-SCL (w10 = 38%) hydrogel are reexamined and compared with those for SofLens One Day and PureVision hydrogels. Measured steady-state water fluxes are largest for SofLens One Day, followed by PureVision and HEMA. In some cases, the measured steady-state water fluxes increase with rising relative air humidity. This increase, due to an apparent mass-transfer resistance at the surface (trapping skinning), is associated with formation of a glassy skin at the air/membrane interface when the relative humidity is below 55-75%. Steady-state water fluxes are interpreted through an extended Maxwell-Stefan diffusion model for a mixture of species starkly different in size. Thermodynamic nonideality is considered through Flory-Rehner polymer-solution theory. Shrinking/swelling is self-consistently modeled by conservation of the total polymer mass. Fitted Maxwell-Stefan diffusivities increase significantly with water concentration in the contact lens.

Biocompatible Materials↗

PubMed2005

Imaging of reconstituted purple membranes by atomic force microscopy.

The organization of bacteriorhodopsin (bR) within reconstituted purple membranes (RPM) was examined using atomic force microscopy (AFM). Five reconstituted species were examined: RPM 3 (bR/native polar lipids/dimyristoylphosphatidylcholine (DMPC) in a 1:9:14 molar ratio), RPM 4 (bR/native polar lipids in a 1:7 molar ratio), RPM 5 (bR/native polar lipids/1,2-di-O-phytanyl-sn-glycerol in a 1:3.5:6.1 molar ratio), RPM 6 (bR/native polar lipids/1,2-di-O-phytanyl-sn-glycero-3-phosphocholine in a 1:3.5:4.9 molar ratio), and RPM 7 (bR/native polar lipids/1,2-diphytanoyl-sn-glycero-3-[phospho-L-serine] in a 1:3.5:4.6 molar ratio). RPM 3 patches adsorbed onto mica exhibit domains of crystallized bR trimers arranged in a hexagonal packing structure, similar to those found in native purple membrane (NPM). These domains are enclosed by DMPC-rich regions. RPM 4 patches were observed to have larger domains of crystallized bR, with trimer orientation 30 degrees different from that found in NPM. The bR-rich domains are enclosed by a large, protein-free, lipid-rich region. The topography of RPM 5 was difficult to resolve as the surface had no discernable patterns or structure. The topographies of RPM 6 and 7 were similar to that found in RPM 3 in that higher domains were formed within the patch adsorbed onto mica. They may contain protein-rich regions, but clear images of protein arrangement could not be obtained using AFM. This may be a result of imaging limitations or of the lack of organization of bR within these domains.

Adsorption↗

PubMed2004

Shear and dilatational relaxation mechanisms of globular and flexible proteins at the hexadecane/water interface.

Proteins adsorbed at fluid/fluid interfaces influence many phenomena: food emulsion and foam stability (Murray et al. Langmuir 2002, 18, 9476 and Borbas et al. Colloids Surf., A 2003, 213, 93), two-phase enzyme catalysis (Cascao-Pereira et al. Biotechnol. Bioeng. 2003, 83, 498; 2002, 78, 595), human lung function (Lunkenheimer et al. Colloids Surf., A 1996, 114, 199; Wustneck et al.; and Banerjee et al. 2000, 15, 14), and cell membrane mechanical properties (Mohandas et al. 1994, 23, 787). Time scales important to these phenomena are broad, necessitating an understanding of the dynamics of biological macromolecules at interfaces. We utilize interfacial shear and dilatational deformations to study the rheology of a globular protein, lysozyme, and a disordered protein, beta-casein, at the hexadecane/water interface. Linear viscoelastic properties are measured using small amplitude oscillatory flow, stress relaxation after a sudden dilatational displacement, and shear creep response to probe the rheological response over broad experimental time scales. Our studies of lysozyme and beta-casein reveal that the interfacial dissipation mechanisms are strongly coupled to changes in the protein structure upon and after adsorption. For beta-casein, the interfacial response is fluidlike in shear deformation and is dominated by interfacial viscous dissipation, particularly at low frequencies. Conversely, the dilatational response of beta-casein is dominated by diffusion dissipation at low frequencies and viscous dissipation at higher frequencies (i.e., when the experimental time scale is faster than the characteristic time for diffusion). For lysozyme in shear deformation, the adsorbed protein layer is primarily elastic with only a weak frequency dependence. Similarly, the interfacial dilatational moduli change very little with frequency. In comparison to beta-casein, the frequency response of lysozyme does not change substantially after washing the protein from the bulk solution. Apparently, it is the irreversibly adsorbed fraction that dominates the dynamic rheological response for lysozyme. Using stress relaxation after a sudden dilatational displacement and shear creep response, the characteristic time of relaxation was found to be 1000 s in both modes of deformation. The very long relaxation time for lysozyme likely results from the formation of a glassy interfacial network. This network develops at high interfacial concentrations where the molecules are highly constrained because of conformation changes that prevent desorption.

Adsorption↗

PubMed2004

Reduced protein adsorption at solid interfaces by sugar excipients.

Sugar excipients are shown to reduce the adsorption of ribonuclease A, bovine serum albumin, and hen egg white lysozyme at the liquid-solid interface. The amount of protein adsorbed decreased as the concentration of the sugar increased. At the same sugar concentration, the ability of sugars to reduce protein adsorption followed the trend: trisaccharides > disaccharides > 6-carbon polyols > monosaccharides. This trend in adsorbed protein amounts among sugars was explained by stabilization of the protein native state in solution by the sugar excipients. The heat of solution of the amorphous saccharide was found to correlate with the amount of protein adsorbed.

Adsorption↗

PubMed2004

Wettability of silicone-hydrogel contact lenses in the presence of tear-film components.

INTRODUCTION: Modern application of soft contact lenses demands safe and comfortable wear over extended time periods up to one month. Lenses that exhibit and sustain complete water wetting allow thick tear-film deposition, minimize film rupture, and permit smooth tear recovery upon lid closure. Water contact angles determined using an air bubble captive on a lens best gauge the in-vivo wetting state. To achieve highly water wetting lenses demands that contact-angle hysteresis be eliminated and that the advancing and receding angles both approach zero. Since lens wear exposes the anterior surface to tear proteins, lens wettability should be measured in the presence of tear-film components. METHODS: A captive-bubble technique is applied to measure the advancing and receding contact angles of two commercial silicone-hydrogel lenses: PureVision (PV) and Focus Night & Day (CF) and a standard HEMA (hydroxethyl-methacrylate) hydrogel lens: Acuvue (AV). In the captive-bubble method, an air bubble immersed in aqueous solution is brought into contact with the contact lens. The contact angle through water during bubble expansion yields the receding angle. Bubble contraction gives the water advancing angle. Contact-angle hysteresis is the difference between the advancing and receding angles. RESULTS: In isotonic solution, all three lenses display considerable contact-angle hysteresis with advancing angles of almost 90 degrees. When lysozyme and/or mucin were added to the aqueous solution, hysteresis was eliminated, and equivalent and high water wetting was achieved for the three lenses. Only the advancing angle in isotonic solution provided discriminating evidence for differences in surface chemistry. Covalent attachment of polyethyleneglygol (PEG) to the PV lens surface achieved complete water wetting independent of the presence of tear protein in the solution. CONCLUSIONS: The captive-bubble technique provides contact angles that are relevant to on-eye lens wear. Both advancing and receding contact angles are important to lens wettability performance. When lysozyme and/or mucin are present in the solution, PV, CF, and AV lenses display low advancing and receding contact angles indicative of equivalent wettability performance. This result is due to molecular adsorption of the proteins onto the lens external surface. Covalently attached PEG on the PV lens not only provides complete water wetting but also minimizes or even eliminates protein adsorption.

Absorption↗

PubMed2004

Solvent-amino acid interaction energies in three-dimensional-lattice Monte Carlo simulations of a model 27-mer protein: Folding thermodynamics and kinetics.

Amino acid residue-solvent interactions are required for lattice Monte Carlo simulations of model proteins in water. In this study, we propose an interaction-energy scale that is based on the interaction scale by Miyazawa and Jernigan. It permits systematic variation of the amino acid-solvent interactions by introducing a contrast parameter for the hydrophobicity, C(s), and a mean attraction parameter for the amino acids, omega. Changes in the interaction energies strongly affect many protein properties. We present an optimized energy parameter set for best representing realistic behavior typical for many proteins (fast folding and high cooperativity for single chains). Our optimal parameters feature a much weaker hydrophobicity contrast and mean attraction than does the original interaction scale. The proposed interaction scale is designed for calculating the behavior of proteins in bulk and at interfaces as a function of solvent characteristics, as well as protein size and sequence.

Algorithms↗

PubMed2003

Interfacial versus homogeneous enzymatic cleavage of mandelonitrile by hydroxynitrile lyase in a biphasic system.

The question of an interfacial versus a homogeneous reaction is carefully addressed for the enzymatic biphasic cleavage of mandelonitrile to benzaldehyde by Prunus amygdalus hydroxynitrile lyase (pa-Hnl) (Hickel et al. [1999] Biotechnol Bioeng 36:425-436). Experimental evidence, including 1) the reaction ceases when the interface is populated by previously adsorbed denatured pa-Hnl, 2) the reaction continues even after washout of the bulk enzyme from the aqueous phase, 3) highly nonpolar organic solvents initially promote fast reaction kinetics that relatively quickly decay to zero product production, and 4) the reaction rate is nonlinear in the bulk enzyme concentration, provide robust grounds for an interfacial reaction. We also model enzymatic mandelonitrile cleavage assuming a homogeneous aqueous-phase reaction. The homogeneous reaction scheme does not simultaneously account for the experimental observations of a linear dependence of the reaction rate on organic/water interfacial area, no dependence on the aqueous-phase volume, and a nonlinear dependence on pa-Hnl aqueous concentration. Further, simple calculations demonstrate that the homogeneous reaction rate is at least three orders of magnitude slower than those observed by Hickel et al. (1999). We again conclude that enzyme adsorbed at the organic solvent/water interface primarily catalyzes the biphasic mandelonitrile cleavage reaction.

Acetonitriles↗

PubMed2003

Fenestrations enhance tear mixing under silicone-hydrogel contact lenses.

PURPOSE: Extended wear of conventional hydrogel soft contact lenses (SCLs) is associated with adverse ocular responses. Some of these ocular events are thought to result from inadequate tear mixing in the postlens tear film (PoLTF). Therefore, strategies to increase tear mixing may improve the safety of extended-wear SCLs. Recently, it has been suggested that placing fenestrations in soft lenses increases tear mixing. In the present study, hydrodynamic modeling and tear-mixing measurements were used to explore the effects of fenestrations on tear exchange under an SCL. METHODS: Tear mixing, expressed as the time to deplete 95% of fluorescent dye from the PoTLF (T(95)), was measured in 20 subjects fitted with two pairs of silicone hydrogel SCLs. The lenses were identical except that one pair was fenestrated (F) with 40 holes each 100 microm in diameter. The effect of fenestrations on improving tear mixing is explained and enhancement of tear mixing is predicted, with the use of hydrodynamic modeling. RESULTS: T(95) estimates were significantly lower (P < 0.001) in the F (mean T(95) = 18.3 minutes) lenses compared with the standard unfenestrated (S-uF) (mean T(95) = 22.6 minutes) lenses (DeltaT(95) = 4.3 minutes, 95% CI = 2.5-6.2) and were in general agreement with the proposed mixing model. Optimization of tear mixing can be achieved by selectively placing fenestrations in the pooling regions before the thinnest regions. The model predictions were sensitive to the distribution of tear-film thickness under the lens. CONCLUSIONS: Fenestrations improve tear-mixing efficiency. Accurate prediction of the effects of lens parameters on tear mixing, however, demands quantitative measurement of the postlens distribution of tear-film thickness.

Adolescent↗

PubMed2002

A kinetic model for enzyme interfacial activity and stability: pa-hydroxynitrile lyase at the diisopropyl ether/water interface.

A kinetic framework is developed to describe enzyme activity and stability in two-phase liquid-liquid systems. In particular, the model is applied to the enzymatic production of benzaldehyde from mandelonitrile by Prunus amygdalus hydroxynitrile lyase (pa-Hnl) adsorbed at the diisopropyl ether (DIPE)/aqueous buffer interface (pH = 5.5). We quantitatively describe our previously obtained experimental kinetic results (Hickel et al., 1999; 2001), and we successfully account for the aqueous-phase enzyme concentration dependence of product formation rates and the observed reaction rates at early times. Multilayer growth explains the early time reversibility of enzyme adsorption at the DIPE/buffer interface observed by both enzyme-activity and dynamic-interfacial-tension washout experiments that replace the aqueous enzyme solution with a buffer solution. The postulated explanation for the unusual stability of pa-Hnl adsorbed at the DIPE/buffer interface is attributed to a two-layer adsorption mechanism. In the first layer, slow conformational change from the native state leads to irreversible attachment and partial loss of catalytic activity. In the second layer, pa-Hnl is reversibly adsorbed without loss in catalytic activity. The measured catalytic activity is the combined effect of the deactivation kinetics of the first layer and of the adsorption kinetics of each layer. For the specific case of pa-Hnl adsorbed at the DIPE/buffer interface, this combined effect is nearly constant for several hours resulting in no apparent loss of catalytic activity. Our proposed kinetic model can be extended to other interfacially active enzymes and other organic solvents. Finally, we indicate how interfacial-tension lag times provide a powerful tool for rational solvent selection and enzyme engineering.

Adsorption↗

PubMed2002

Black-line formation and the "perched" human tear film.

Introduction. McDonald and Brubaker(1) observed that after instillation of fluorescein, dark lines appear near the upper and lower lid margins of the human eye immediately following a blink. Methods. This paper numerically models the dynamic black-line formation under the lubrication approximation at low Reynolds number, including gravity and surface tension forces. Results. The hydrodynamic model predicts that menisci at the lid margins draw liquid from the tear film, resulting in local thinning close to the lids (i.e., black lines) due to capillary suction. Once the film thins locally, resistance to further thinning increases. No vertical gravity-induced drainage occurs between blinks, and the tear film is trapped in its initially deposited state or "perched" between the two black lines. Conclusions. We demonstrate quantitatively that due to rapid formation of black lines, the tear film is trapped for extended periods. Lid action is therefore required to disturb the perched tear film.

Adult↗