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PubMed1993

Optimal test parameters for computerized quantitative layer-by-layer perimetry.

Quantitative layer-by-layer perimetry is a psychophysical technique believed to assess the functional characteristics of individual retinal layers. In the initial design of the technique, the test procedure was too time consuming for the testing of large field point patterns, and statistical treatment of the measured threshold values was difficult. Therefore, to minimize these limitations, a new design of quantitative layer-by-layer perimetry was developed by incorporating the technique into the existing hardware environment of the computerized perimeter Humphrey Field Analyzer. The present paper describes the theoretical considerations and experimental findings underlying a selection of optimal stimulus parameters to provide a basis for statistical treatment of measured threshold values, and to minimize test duration, while obtaining an acceptable estimate of the threshold level with minimum variance.

Adult↗

PubMed2005

Controlled release of dexamethasone from microcapsules produced by polyelectrolyte layer-by-layer nanoassembly.

PURPOSE: In an effort to expand the application of core-shell structures fabricated by electrostatic layer-by-layer (LbL) self-assembling for drug delivery, this study reports the controlled release of dexamethasone from microcrystals encapsulated with a polyelectrolyte shell. METHODS: The LbL self-assembly process was used to produce dexamethasone particles encapsulated with up to five double layers formed by alternating the adsorption of positively charged poly(dimethyldiallyl ammonium chloride), negatively charged sodium poly(styrenesulfonate) and depending on the pH positively or negatively charged gelatin A or B onto the surface of the negatively charged dexamethasone particles. The nano-thin shells were characterized by quartz crystal microbalance measurements, microelectrophoresis, microcalorimetry, confocal microscopy, and scanning electron microscopy. In vitro release of dexamethasone from the microcapsules suspended in water or carboxymethylcellulose gels were measured using vertical Franz-type diffusion cells. RESULTS: Sonication of a suspension of negatively charged dexamethasone microcrystals in a solution of PDDA not only reduced aggregation but also reduced the size of the sub-micrometer particles. Assembly of multiple polyelectrolyte layers around these monodispersed cores produced a polyelectrolyte multilayer shell around the drug microcrystals that allowed for controlled release depending on the composition and the number of layers. CONCLUSIONS: Direct surface modification of dexamethasone microcrystals via the LbL process produced monodispersed suspensions with diffusion-controlled sustained drug release via the polyelectrolyte multilayer shell.

Capsules↗

PubMed1975

Isolation, ultrastructure and chemical composition of the outermost layer ("exo-layer") of the Epidermophyton floccosum cell wall.

The outer-most layer ("exo-layer") of the wall was isolated from cell walls of Epidermophyton floccosum. The pure cell walls, obtained by disruption in a Ribi cell fractionator, sonication and centrifugation, were digested with snail enzyme for 12 h. Thereafter, the exo-layer preparation was obtained as the fraction resistant to the snail enzyme. Electron microscopy showed that the exo-layer is a thin, stranded network structure 10-20 nm thick. Chemical analysis of the exo-layer showed that the main components are protein (63 percent), mannose (10 percent) and glucosamine (17 percent). Sodium dodecyl sulfate polyacrylamide gel electrophoresis has revealed that the main band is a glycoprotein containing mannose.

Amino Acids↗

PubMed1986

Three-dimensional structure of the regular surface layer (HPI layer) of Deinococcus radiodurans.

The low-resolution structure of the regular surface layer of Deinococcus radiodurans has been determined from negatively stained specimens by three-dimensional electron microscopy. The layer has P6 symmetry, a lattice constant of 18 nm and a thickness of 6.5 nm. Three-dimensional reconstruction was performed by a hybrid real space/Fourier space approach that incorporates partial compensation of lattice distortions: The model obtained is discussed in the light of independent information about the surface structure of this layer, derived from metal shadowing and surface relief reconstruction. While agreement is quite satisfactory for the apparently more rigid inner surface, the outer surface shows severe flattening effects. The structure of the HPI layer is compared with other bacterial surface layers using a classification scheme that is outlined in the Appendix.

Bacteria↗

PubMed1993

Induction of T-cell immunity to oligosaccharide antigens immobilized on crystalline bacterial surface layers (S-layers).

Immunization of Balb/c mice with conjugates of oligosaccharide haptens and crystalline bacterial surface-layer proteins (S-layers) primed the mice for a strong, hapten-specific, delayed-type hypersensitivity (DTH) response. Conjugates of haptens with bovine serum albumin produced only weak DTH responses but, when mixed with aluminium hydroxide, elicited DTH responses comparable to those against S-layer conjugates. Surface-layer conjugates also elicited strong anti-hapten DTH responses when administered by an oral/nasal route. Apparently, the natural assembly of S-layer proteins into large, two-dimensional arrays endows them with intrinsic adjuvant properties.

Adjuvants, Immunologic↗

PubMed1996

Biotechnology and biomimetic with crystalline bacterial cell surface layers (S-layers).

Crystalline bacterial cell surface layers (S-layers) are the outermost cell envelope component of many eubacteria and archaeobacteria. S-layers are composed of a single protein or glycoprotein species and exhibit oblique, square or hexagonal lattice symmetry. Pores passing through these monomolecular arrays show identical size and morphology, and functional groups are aligned in well-defined positions and orientations. Due to these unique features, S-layers have broad application potential in biotechnology including functioning as biomimetic membranes. Presently, S-layers are used (i) for the production of isoporous ultrafiltration membranes with very well defined molecular sieving and adsorption properties, (ii) as matrices for the controlled immobilization of biologically active macromolecules (e.g., enzymes, antibodies, ligands) as required for biosensors, affinity membranes and affinity microparticles as well as for solid phase assays, (iii) as stabilizing structures for Langmuir-Blodgett films and liposomes and (iv) as carriers and adjuvants for weakly immunogenic antigens and haptens.

Adjuvants, Immunologic↗

PubMed2004

Engineered microcrystals for direct surface modification with layer-by-layer technique for optimized dissolution.

This investigation relates to a two-step formulation development technique-synthesis of sterically stabilized drug microcrystals followed by direct surface modification by sequential electrostatic adsorption. Stable microcrystals of naproxen were produced by pH-induced reprecipitation in presence of a stabilizer. Sequential layer growth was achieved by the layer-by-layer assembly of biocompatible polyelectrolytes (PEs) and was registered using microelectrophoresis. The coated colloids were characterized using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). The in vitro controlled release pattern of the drug through the PE diffusion barrier was studied using a diffusion cell assembly at physiological pH of 7.4, both before and after freeze-drying. Thermodynamically stable naproxen microcrystals were obtained by association and had a mean length of 15 microm and a zeta potential of -37.5 mV and were surface modified efficiently using biocompatible polysaccharide/protein-based PEs. Sufficient charge reversal with each layer was evident indicating layer growth with successive deposition cycles. The coating was complete and homogeneous as visualized under CLSM and SEM. The in vitro release study revealed that the stoichiometry of PEs in the complex coating and its molecular architecture played important roles in forming the diffusion barrier, which offered efficient control of the dissolution rate of drug core (up to 50% lower than bare crystal). The release profile fitted zero order release kinetics. This novel formulation technique enables administration of high concentrations of water-insoluble drugs in a stable, tissue compatible form, simultaneously affording sustained release.

Chemical Engineering↗

PubMed2006

Layer-by-layer assembly of intact polydiacetylene vesicles with retained chromic properties.

Photopolymerized vesicles of 10,12-pentacosadiynoic acid (PPCDA) were successfully assembled into polyelectrolyte multilayer (PEM) thin films using either chitosan or poly(ethylenimine) (PEI) as a polycation. The PEM films assembled from chitosan or PEI polycation retained the blue color of PPCDA vesicles. For the blue films, an increase in absorbance at 635 nm as a function of the number of deposited layers of PPCDA vesicles was observed, confirming the uniform layer-by-layer deposition process. The spherical structures of PPCDA vesicles, as well as their important colorimetric responses to solvent (ethanol), pH, and temperature, are retained in the PEM film. Compared to the vesicles dispersed in water, the PEM films are much more stable to aging. Layer-by-layer assembly thus provides a convenient means to prepare colorimetric sensing devices, with extended shelf life, from polydiacetylene vesicles.

Journal Article↗

PubMed2003

Layer-by-layer self-assembly of Prussian blue colloids.

The adsorption of Prussian blue (PB) colloids within layers of polyelectrolytes has been achieved by a reiterative immersion-rinse approach. Multilayer assemblies consisting of alternate layers of these components have been prepared by the layer-by-layer (LbL) self-assembly technique. Both processes have been carefully monitored by cyclic voltammetry and infrared and UV-visible spectroscopy. Linear increase in the IR and UV-visible light absorbance with the number of deposited layers indicates that well-organized lamellar systems have been elaborated. Size and distribution of Prussian blue nanoparticles in these systems have been investigated by AFM. The effect of the molar concentration of the PB dipping solution on the adsorption process and the distribution of the PB colloids has also been described. Finally, magnetic properties of these assemblies have been studied by low-temperature ESR measurements. Indeed, this new approach of hybrid LbL films opens the way to a new class of nanostructured lamellar compounds.

Adsorption↗

PubMed1997

Polymerization contraction stress in thin resin composite layers as a function of layer thickness.

OBJECTIVES: In the present study, the effect of layer thickness on the curing stress in thin resin composite layers was investigated. Since the value of the contraction stress is dependent on the compliance of the measuring equipment (especially for thin films), a method to determine the compliance of the test apparatus was tested. METHODS: A chemically initiated resin composite (Clearfil F2, Kuraray) was inserted between two sandblasted and silane-coated stainless steel discs in a tensilometer. The curing contraction of the cylindrical samples was continuously counteracted by feedback displacement of the tensilometer crosshead, and the curing stress development was registered. After 20 min, the samples were loaded in tension until fracture. The curing stress was determined for layer thicknesses of 50, 100, 200, 300, 400, 500, 600, 700 microns, 1.4 mm and 2.7 mm. The compliance of the apparatus was calculated with the aid of a non-linear regression analysis, using an equation derived from Hooke's Law as the model. RESULTS: None of the samples fractured due to contraction stress prior to tensile loading. The contraction stress after 20 min decreased from 23.3 +/- 5.3 MPa for the 50 microns layer to 5.5 +/- 0.6 MPa for the 2.7 mm layer. The compliance on the apparatus was 0.029 mm/MPa. SIGNIFICANCE: A measuring method was developed which was found to be suitable for the determination of axial polymerization contraction stress in this films of chemically initiated resin composites. The method makes it possible to estimate the stress levels that occur in resin composite films in the clinical situation.

Analysis of Variance↗

PubMed2002

Parabrachio-cortical connections with the lateral hemisphere in the madagascan hedgehog tenrec: prominent projections to layer 1, weak projections from layer 6.

The present study was undertaken to further characterize and subdivide the rhinal cortex (insular and perirhinal areas) in the hedgehog tenrec (Echinops telfairi), a placental mammal with a rather low encephalisation index. Injections of wheat germ agglutinin-horseradish peroxidase into the dorsolateral pontine tegmentum revealed a prominent layer 1 projection to several rhinal target areas, while the rhinal cortex only stained weakly for the calcitonin gene-related peptide. Among the regions retrogradely labeled following tracer injections into the rhinal cortex, the parabrachial nucleus was considered the main origin of the tegmento-cortical projection. This conclusion was based on the circumscribed pattern of termination, as well as the differences noted between the pattern of anterograde labeling and the pattern obtained by thyrosine hydroxylase immunohistochemistry. The tracer injections into the dorsolateral tegmentum also revealed numerous retrogradely labeled cells in the layer 5 of the dorsomedial frontal cortex. In contrast, the rhinal cortex only showed few labeled cells and most of these cells were located in the layer 6/7. A comparison with other species indicates that the tenrec's parabrachial nucleus gives rise to the most extensive cortical projections but receives the least prominent input from the lateral cerebral hemisphere. The layer 6/7 projection may be a common mammalian feature but it is overshadowed by the layer 5 projection in higher mammals.

Animals↗

PubMed2002

Influence of layer position on in vitro and in vivo release of levodopa methyl ester and carbidopa from three-layer matrix tablets.

A versatile oral controlled release system for the simultaneous delivery of levodopa methyl ester and carbidopa, consisting of a three-layer matrix tablet, has been studied and developed. Each individual layer of the matrix exhibited a different release mechanism, i.e. the first layer was swellable (S), the second one was erodible (E) and the third one was disintegrating (D). The three layers have been assembled in the monolithic matrix in different relative positions. It was found that in the monolith the three layers could interact, producing in vitro release profiles depending on their relative position. The monoliths having the configurations DSE and SDE were administered to human volunteers in order to determine the plasma profiles. The pharmacokinetic data showed a significant difference between the early time plasma curves: the monolith DSE, having the fast release profile, gave rise to a rapid appearance of a high levodopa plasma level, whereas the slower releasing monolith SDE produced a smoothed plasma concentration profile.

Administration, Oral↗

PubMed2005

A reactive poly(ethylene glycol) layer to achieve specific surface plasmon resonance sensing with a high S/N ratio: the substantial role of a short underbrushed PEG layer in minimizing nonspecific adsorption.

A reactive poly(ethylene glycol) (PEG)-brushed layer was constructed on a surface plasmon resonance (SPR) sensor chip using a heterobifunctional PEG possessing an acetal group at one end and a mercapto group at the other end (alpha-acetal-omega-mercapto-PEG). The density of the PEG brushed layer substantially increased with repetitive adsorption/rinse cycles of the PEG on the sensor chip, allowing dramatic reduction of nonspecific protein adsorption. Notably, formation of a short, filler layer of PEG (2 kDa) in the preconstructed longer PEG brushed layer (5 kDa) achieved almost complete prevention of nonspecific protein adsorption. The acetal group located at the distal end of the tethered PEG was converted to an aldehyde group by the acid treatment, followed by the installation of biocytin hydrazide through Schiff base formation. SPR sensing of streptavidin was done with a very high S/N ratio even in a proteinous medium using the biotinylated PEG (5 kDa) tethered chip with an inert filler layer of short PEG (2 kDa). Furthermore, the specific affinity of streptavidin for the biotinylated PEG was highly influenced by the length of the filler PEG and was significantly reduced when the length of the filler PEG was longer than that of the biotinylated PEG. This result clearly revealed the substantial importance of the steric factor on biospecific interaction at the distal end of tethered PEG on the sensor surface.

Adsorption↗

PubMedDate not supplied

Alternating bioactivity of polymeric layer-by-layer assemblies: anticoagulation vs procoagulation of human blood.

The layer-by-layer assembly between cationic chitosan and anionic dextran sulfate was analyzed quantitatively by a quartz crystal microbalance technique in the absence and presence of 0.2, 0.5, and 1 M NaCl in the polymer solution. The apparent film thickness increased upon increasing the NaCl concentration. The anti- versus procoagulant activity of these films against whole human blood was studied by the immersion of a substrate into blood for 30 min incubation time at 37 degrees C. The substrate was coated with films of varying NaCl concentrations and assembly step numbers. There was a critical concentration for the alternating activity; above a concentration of 0.5 M NaCl, both anti- and procoagulation could be observed on the dextran sulfate and chitosan surfaces, respectively. The underlying layer of the assembly was necessary for this alternating activity; after a five-step assembly, the activity was realized. The adsorption of a cationic dye (methylene blue) onto the films revealed that the anionic-charge density derived from dextran sulfate on the film surface was linearly increased with increased NaCl concentration. There was a critical charge density of the dextran sulfate for the anticoagulant activity. An assembly was also constructed from a combination of chitosan and heparin, but the activity was different from that of the former system; strong anticoagulant activity was observed even on the chitosan surface. We suggest that the polymer species and/or the assembly conditions are key factors for realizing the alternating bioactivities of films prepared by the layer-by-layer assembly.

Blood Coagulation↗

PubMed2006

Electrochemically induced disintegration of layer-by-layer-assembled thin films composed of 2-iminobiotin-labeled poly(ethyleneimine) and avidin.

A layer-by-layer assembly composed of avidin and 2-iminobiotin-labeled poly(ethyleneimine) (ib-PEI) was prepared on the surface of a platinum (Pt) film-coated quartz resonator, and an electrochemically induced disintegration of the avidin-ib-PEI assembly was studied using a quartz crystal microbalance. The resonance frequency of a five-bilayer (avidin-ib-PEI)5 film-coated quartz resonator was increased upon application of an electric potential to the Pt layer of the quartz resonator, suggesting that the mass on the quartz resonator was decreased as a result of disintegration of the (avidin-ib-PEI)5 film, due to a pH change in the vicinity of the surface of the Pt-coated quartz resonator. It may be that the (avidin-ib-PEI)5 film assembly was decomposed by acidification of the local pH on the surface of the Pt layer, which in turn was induced through electrolysis of water on Pt, because ib-PEI forms complexes with avidin only in basic media. In pH 9 solution, the (avidin-ib-PEI)5 film was decomposed under the influence of an applied potential of 0.6-1.0 V versus Ag/AgCl. The (avidin-ib-PEI)5 film was decomposed almost completely within a minute in a low concentration buffer (1 mM, pH 9), while the decomposition was slower in 10 and 100 mM buffer solutions at the same pH. The decomposition of the assembly was rapid when the electrode potential was applied in pH 9 solutions, while the response was relatively slow in pH 10 and 11 solutions. All the results are rationalized on the basis of an electrochemically induced acidification of the local environment around the (avidin-ib-PEI)5 film on the Pt layer.

Avidin↗

PubMed2006

Construction of 3D layer-pillared homoligand coordination polymers from a 2D layered precursor.

Herein, we present a new method for preparing homoligand 3D coordination polymers. First, a layered metal-organic framework Zn3(BDC)3(H2O)2 x 4DMF 1 (BDC is terephthalate, DMF is N,N-dimethylformamide) was fabricated from a H2BDC by liquid-liquid diffusion. Second, the layered product, 1, was used as a precursor to solvothermally react with further H2BDC at 140-180 degrees C, resulting in two products of BDC insertion into the layered structure. These are [Zn3(p-BDC)4] x 2HPIP, 2 (HPIP is partly protonated piperazine), and [Zn3(p-BDC)3(H2BDC)] x (C6H15NO) x H2O x 3DMF, 3 (C6H15NO is triethylamine N-oxide). Single-crystal X-ray diffraction shows that 2 possesses a layer-pillared structure of mu4-BDC, with 1D channels, while 3 has a layer-pillared structure of mu2-BDC, with 2D channels. N2-sorption experiments show 3 has a relatively high BET surface area of 750 m(2)/g. It is proposed that 2 follows the crystal growth mechanism of Ostwald ripening, whereas the crystal structure of 3 might be formed by an insertion mechanism.

Journal Article↗

PubMed2004

Layer-by-layer nanotube template synthesis.

Electroless deposition of gold on the pore walls of polycarbonate templates is currently the best known method for controlling inside diameters of template-synthesized nanotubes. It would be very useful to have alternative template-based synthetic chemistries that yield nanotubes composed of other materials, but which still allow for precise control over the nanotube wall thickness and i.d. A film-formation process that is based on layer-by-layer deposition of the film-forming material along the pore walls of the template membrane provides this desired alternative synthetic chemistry. We describe here the use of Mallouk's alpha,omega-diorganophosphonate/Zr layer-by-layer film-forming method for preparing nanotubes within the pores of alumina template membranes. We have found that this method allows accurate, quantitative, and predictable control over the wall thickness, and thus i.d., of the layered nanotubes obtained.

Journal Article↗

PubMed2006

Controlling structure from the bottom-up: structural and optical properties of layer-by-layer assembled palladium coordination-based multilayers.

Layer-by-layer assembly of two palladium coordination-based multilayers on silicon and glass substrates is presented. The new assemblies consist of rigid-rod chromophores connected by terminal pyridine moieties to palladium centers. Both colloidal palladium and PdCl2(PhCN)2 were used in order to determine the effect of the metal complex precursor on multilayer structure and optical properties. The multilayers were formed by an iterative wet-chemical deposition process at room temperature in air on a siloxane-based template layer. Twelve consecutive deposition steps have been demonstrated resulting in structurally regular assemblies with an equal amount of chromophore and palladium added in each molecular bilayer. The optical intensity characteristics of the metal-organic films are clearly a function of the palladium precursor employed. The colloid-based system has a UV-vis absorption maximum an order of magnitude stronger than that of the PdCl2-based multilayer. The absorption maximum of the PdCl2-based film exhibits a significant red shift of 23 nm with the addition of 12 layers. Remarkably, the structure and physiochemical properties of the submicron scale PdCl2-based structures are determined by the configuration of the approximately 15 angstroms thick template layer. The refractive index of the PdCl2-based film was determined by spectroscopic ellipsometry. Well-defined three-dimensional structures, with a dimension of 5 microm, were obtained using photopatterned template monolayers. The properties and microstructure of the films were studied by UV-vis spectroscopy, spectroscopic ellipsometry, atomic force microscopy (AFM), X-ray reflectivity (XRR), scanning electron microscopy (SEM), and aqueous contact angle measurements (CA).

Journal Article↗