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At least 487 records · Page 27Linked to original sources

Polystyrene film-coated glassware: a new means of reducing metal losses in trace metal speciation.

A recently developed process for coating a glass surface with polystyrene (PS) film, by use of a simple chemical process has been used to reduce trace metal adsorption by cell components. The glass coating is a two-step procedure consisting of covalent attachment of vinyl-terminated PS to Si atoms on the glass surface then adsorption of PS from solution to create a stable PS film. To assess the quality of the coating we used anodic stripping voltammetry to study the adsorption of lead and cadmium ions in coated and untreated glass cells. In both short and long-term (24 h) experiments we observed that the amount of metal adsorbed was considerably less for the PS film-coated glass cell than for the uncoated cell. Further experiments showed that metal desorption is faster and metal contamination after cleaning is significantly lower for the coated cells. The PS film was, moreover, stable over a period of 6 months within the pH range 3.5-9.

Adsorption↗

A facile method for coding and labeling assays on polystyrene beads with differently colored luminescent nanocrystals.

The present paper describes a new, facile and fast technique that can be used to incorporate CdSe/ZnS core/shell nanocrystals into polystyrene beads. The nanocrystals were immobilized within the polymer beads by demixing two nonmiscible solvents. The resulting beads were bright, homogeneously coded, and did not show any surface texture or photobleaching under application-relevant conditions. The beads were subsequently used on a model streptavidin-biotin binding system in order to demonstrate the applicability of the technique to enzyme-linked assays and multiplexing applications.

Biotin↗

A polystyrene based membrane electrode for cadmium(II) ions.

A polystyrene based membrane of 3,4:12,13-dibenzo-2,5,11,14-tetraoxo- 1,6,10,15-tetraazacyclooctade-cane shows a Nernstian response to Cd(II) ions over a wide concentration range (3.16 x 10(-6) - 1.00 x 10(-1) mol L(-1) with a Nernstian slope of 29.8 mV/decade of concentration, between pH 2.0 and 6.0. This electrode has been found to be chemically inert and of adequate stability with a response time of 20s. The electrode gives reproducible results with a lifetime of 130 days. The membrane works satisfactorily in a partially non-aqueous medium up to a maximum 35% (v/v) content of methanol and ethanol. The practical utility of the proposed chemical sensor has been observed by using it as end-point indicator in the titration of Cd(II) ions with EDTA. The potentiometric selectivity coefficient values indicate that the membrane sensor is highly selective for Cd(II) ions over a number of cations. Small amounts of surfactants do not disturb the functioning of the sensor. This electrode has also been used to estimate cadmium ions in real samples.

Cadmium↗

An immunoassay for terbutryn using direct hapten linkage to a glutaraldehyde network on the polystyrene surface of standard microtiter plates.

2-Aminobutylamino-4-ethylamino-6-isopropylamino-1,3,5-triazine (ABA-atrazine) has been synthesized and used as a coating hapten in an immunoassay with a monoclonal antibody against terbutryn. Coating was achieved by covalently linking ABA-atrazine to a glutaraldehyde polymer network directly bound to the polystyrene surface of a standard 96-well microtiter plate. The assay was carefully optimized. In particular, the coating hapten concentration had a strong effect on the ELISA sensitivity. By including a pre-incubation step a low test midpoint (IC50-value) of 0.130 microg L(-1) was achieved. As far as we are aware this is the most sensitive ELISA for terbutryn yet reported. The coating-hapten-format presented is proposed as generally applicable, because the glutaraldehyde-modified microtiter plate surface enables stable immobilization of a broad variety of coating haptens.

Antibodies, Monoclonal↗

Development and characterization of bioadhesive vaginal films of sodium polystyrene sulfonate (PSS), a novel contraceptive antimicrobial agent.

PURPOSE: Polystyrene sulfonate (PSS) is a novel noncytotoxic antimicrobial contraceptive agent. A gel formulation of PSS was found safe for vaginal administration in phase I clinical trials. The purpose of the current study was to develop and evaluate novel bioadhesive vaginal film formulations of PSS. METHODS: PSS films were prepared by solvent evaporation and optimized for various physical, mechanical, and aesthetic properties. Further, films were evaluated for various biological activities and safety. RESULTS: Vaginal films containing 300 mg PSS per unit have been developed, using generally regarded as safe (GRAS) listed excipients. The films are colorless, transparent, thin, soft, and tough, dissolve rapidly in physiologic fluid to form a smooth, viscous and bioadhesive solution that could be retained in the vagina for prolonged intervals. Sperm function inhibition (hyaluronidase and cervical mucus penetration) and antimicrobial activities against human immunodeficiency virus (HIV) and herpes simplex virus (HSV) by PSS films were found comparable to PSS. Also, films did not inhibit normal vaginal microflora (Lactobacillus) and were noncytotoxic as indicated by negligible sperm immobilization and cytotoxicity to host cell assays. CONCLUSIONS: Rapidly dissolving bioadhesive vaginal film formulation of PSS with desired physical, mechanical, aesthetic, and biological properties is a suitable candidate vaginal microbicide for prevention of sexually transmitted disease (STDs) and is ready for toxicological and clinical evaluation.

Animals↗

Interaction of antibody with antigen immobilized on polystyrene latex beads: characterization by density gradient centrifugation.

Isopycnic banding by density gradient centrifugation was used to measure density changes in complexes formed by the interaction between antigen and antibody immobilized on polystyrene latex beads (diameter, 0.109 +/- 0.0025 micron). Measurements of density changes allowed calculation of the interacting masses under the given experimental conditions. Interaction equilibrium constants and free energy change for two sets of reactions, bovine IgG and anti-bovine IgG (rabbit) IgG and rabbit IgG and anti-rabbit IgG (goat) IgG systems, were calculated from isopycnic banding density gradient centrifugation runs. The procedure demonstrates a new method of obtaining quantitative information on antigen-antibody interactions.

Animals↗

Protein quantitation of as low as 10-ng/ml concentrations by competitive binding to polystyrene latexes.

A protein quantitation method which offers protein detection as low as 10 ng protein/ml and accurate quantitation as low as 30-100 ng protein/ml, depending on the protein, has been designed. The assay, which is relatively quick and simple to perform, utilizes the strong, nonspecific adsorption of proteins onto polystyrene latexes. A competition is created between a marker enzyme and the analyte protein for a limited amount of latex surface area. Due to inactivation of the enzyme upon binding to a hydrophobic latex surface, measurement of enzyme activity allows determination of the bound/free enzyme ratio and thus the competing protein concentration. Considerations of sensitivity and simplicity are suggested to make this assay superior to others presently available.

Alkaline Phosphatase↗

Determination of the affinity of vitamin D metabolites to serum vitamin D binding protein using assay employing lipid-coated polystyrene beads.

We have developed an assay to measure the affinity of serum vitamin D binding protein for 25-hydroxyvitamin D3, 1,25-dihydroxyvitamin D3, and vitamin D3, using uniform diameter (6.4 microns) polystyrene beads coated with phosphatidylcholine and vitamin D metabolites as the vitamin D donor. The lipid metabolite coated beads have a solid core, and thus all of the vitamin D metabolites are on the bead surface from which transfer to protein occurs. After incubating these beads in neutral buffer for 3 h, essentially no 3H-labeled vitamin D metabolites desorb from this surface. Phosphatidylcholine/vitamin D metabolite-coated beads (1 microM vitamin D metabolite) were incubated with varying concentrations of serum vitamin D binding protein under conditions in which the bead surfaces were saturated with protein, but most of the protein was free in solution. After incubation, beads were rapidly centrifuged without disturbing the equilibrium of binding and vitamin D metabolite bound to sDBP in solution was assayed in the supernatant. All three vitamin D metabolites became bound to serum vitamin D binding protein, and after 10 min of incubation the transfer of the metabolites to serum vitamin D binding protein was time independent. The transfer followed a Langmuir isotherm, and the Kd for each metabolite binding to serum vitamin D binding protein was derived by nonlinear least-squares fit analysis. From this analysis the following values for the Kd were obtained: 5.59 x 10(-6) M, 25-hydroxyvitamin D; 9.45 x 10(-6) M, 1,25-dihydroxyvitamin D; and 9.17 x 10(-5) M, vitamin D. In conclusion, we have developed a method which avoids problems encountered in previous assays and allows the precise and convenient determination of binding affinities of vitamin D metabolites and serum vitamin D binding protein.

Animals↗

A quantitative microassay of carbohydrate-mediated cell adhesion to glycoconjugates immobilized on polystyrene plates.

A microadhesion assay that allows the quantitative determination of carbohydrate-mediated cell adhesion to glycoconjugates immobilized on 96-well polystyrene plates has been developed. After dislodging nonadherent cells by centrifugation, specifically bound cells are quantified by colorimetric analysis of a blue formazan product generated from the dye 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide by enzymatic reduction. Carbohydrate specificity of the cell adhesion was demonstrated by inhibition analyses and the general applicability of the assay was proved with indicator cells of three different origins: mouse fibrosarcoma cells, Chang liver cells, and human breast carcinoma cells (MDA-MB 231).

Animals↗

Catalytic properties and electrostatic potential of charged immobilized enzyme derivatives. Pyruvate decarboxylase attached to cationic polystyrene beads of different charge densities.

Pyruvate decarboxylase has been covalently attached to positively charged macroporous polystyrene resins. The changes in the functional behaviour of the enzyme resulting from the electrostatic interaction of the cationic carrier polymer and the anionic substrate have been studied as function of the fixed charge concentration of the carrier. The findings confirm the connection of two widely used theoretical expressions relating electrostatic parameters with Michaelis constants.

Carboxy-Lyases↗

Receptor-mediated formation of multilayer aggregates of primary cultured adult rat hepatocytes on lactose-substituted polystyrene.

We used a lactose-substituted polystyrene, poly-N-p-vinylbenzyl-D-lactonamide (PVLA), as a substratum for adult rat hepatocytes in primary culture. Spherical-shaped hepatocytes attached on PVLA substratum formed stable multilayer aggregates anchored on substratum through the stimulation of epidermal growth factor (EGF). The cells required calcium ion essentially to form the aggregates. The formation of multilayer aggregates was inhibited by colchicine, but not by cytochalasin B. The inhibition was also observed by added PVLA molecules in the culture medium and by treating surfaces of PVLA-coated dishes with allo A lectin. It was suggested that adult rat hepatocytes attached on PVLA substratum required the specific interaction between asialoglycoprotein receptors on the cell surface and PVLA substratum to form anchored multilayer aggregates.

Animals↗

Visualization of migration of transplanted astrocytes using polystyrene microspheres.

Fluorescent polystyrene microspheres have been used to mark rat astrocytes for transplantation. Highly fluorescent cells were generated by incorporation of microspheres over periods of the order of 10 h. Microspheres ranging in average diameter from 50 to 200 nm and derivatized with either rhodamine or fluorescein have been used to label cells. There were no apparent cytotoxic effects of microsphere incorporation. Loaded astrocytes retained their complement of microspheres over periods of at least several weeks. The microspheres could be visualized in the transmission electron microscope. They were most likely phagocytosed by a coated-pit mechanism and accumulated in various types of lysosomal vesicles. Labelled astrocytes from neonatal PVG rats were implanted stereotaxically (as a plasma clot) into hippocampi of recipient syngeneic adult rats. Donor cells migrated from transplant to host over about 7 days. Transplant cells tended to be close to blood vessels over this period. Their location in paravascular spaces was determined from observation of microsphere-bearing cells in electron microscopy. Microsphere-loading of astrocytes is a simple method for marking cells for extended periods of time for transplantation which allows direct identification of marked cells with both fluorescence and electron microscopes.

Animals↗

High-performance affinity chromatography of human thrombin on modified polystyrene resins.

Cross-linked polystyrenes modified with L-arginyl methyl ester mimic the binding site of antithrombin III and thrombin substrates. They can be used as stationary phases in high-performance affinity chromatography of thrombin. Under isocratic conditions, thrombin is strongly adsorbed on the resins when the sodium chloride concentration is lower than 0.5 M. The bound enzyme can be selectively desorbed when the salt concentration is raised to about 1.2 M. With a linear salt gradient, the specific elution of thrombin can be effected with a high recovery of its enzymatic activity. The decomposition products of thrombin, when treated with sodium dodecyl sulphate, are not retained by the stationary phase. The effects of the flow-rate and salt gradient slope on the adsorption and desorption of alpha-thrombin demonstrate the importance of kinetic parameters.

Adsorption↗

Adsorption behavior of milk proteins on polystyrene latex. A study based on sedimentation field-flow fractionation and dynamic light scattering.

Sedimentation field-flow fractionation (SdFFF) has been used to characterize the adsorption of the proteins beta-casein (BCN) or beta-lactoglobulin (BLG) on colloidal polystyrene latices; this system was used to model hydrophobic interactions between the proteins and the surfaces of fat droplets in protein-stabilized emulsions. It was found that the SdFFF technique could determine directly the surface concentrations of BCN and BLG irreversibly adsorbed to the latex surface, provided care was taken to maintain the ionic strength of the carrier at a level which suppressed particle-wall repulsion in the separation channel. The measured surface concentrations were similar for the two proteins (about 1 mg/m2), and this was verified by quantitative amino acid analysis. These concentrations were smaller than those found in depletion studies (3 and 4 mg/m2 respectively for BCN and BLG), in which loosely associated protein may have been included in the determinations. The thickness of the adsorbed layers was determined in situ by dynamic light scattering and was found to differ significantly for the two proteins (up to 15 nm for BCN vs. 2-3 nm for BLG). The implication of these findings in terms of different surface arrangements of the two proteins is discussed.

Calorimetry↗

A micromethod for evaluating the phagocytic activity of human macrophages by ingestion of radio-labelled polystyrene particles.

Studies in vitro of human macrophage function in health and disease have been impeded by the difficulty of obtaining such cells in sufficient number. Unlike animal species, the only readily available source of human macrophages are circulating monocytes. Herein, a method is described whereby the phagocytic rate of small numbers of glass-adherent mononuclear cells can be accurately measured. The method utilizes the ingestion by macrophages of technetium labelled polystyrene particles; both the radiolabel and ingestible substrate are readily available and the labelling process simple and efficient. The phagocytic rate can be expressed as radioactive counts per microgram of cell protein; data is also presented showing that the number of particles ingested per cell can be accurately derived.

Blood Cell Count↗

Cross-linking of Schistosoma mansoni antigens and their covalent binding on the surface of polystyrene microtitration trays for use in the ELISA.

The usefulness of the enzyme-linked immunosorbent assay (ELISA) for serodiagnosis is limited by the incomplete and unequal binding of individual antigens present in a complex antigen preparation. We therefore investigated 2 modifications of the commonly used antigen coating procedure in the ELISA. Schistosoma mansoni antigens cross-linked by N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide (EDC), and then adsorbed on polystyrene microtitration trays, tested with sera from patients, and from immunized rabbits, showed an increase in the ELISA results, with respect to both absorbance values and titers. Measurement of specific antibodies against low molecular weight schistosomal antigens, which is not possible in the standard ELISA, was made possible by adsorption of a cross-linked antigen preparation. Covalent binding of cross-linked antigens to modified microtitration trays, gave a further increase in ELISA readings when the antibody response of S. mansoni infected mice to lower molecular weight antigens was assayed. In assays of patients' sera, a further increase in sensitivity of the ELISA was occasionally obtained by binding cross-linked antigens covalently. With some sera the use of physically adsorbed cross-linked antigens gave optimal results.

Animals↗

A new microtechnique for quantitating cell movement in vitro using polystyrene bead monolayers.

In this paper we describe a new and simple technique for studying cell movement in vitro. This method is based upon the migration of endothelial cells on polystyrene bead monolayers. It permits the rapid preparation and screening of multiple 96-well plates for effects on migration, requires fewer samples per test and has the potential for precise quantitation by digital image analysis. The locomotive response of endothelial cells in response to different protein matrices (fibrin, collagen, albumin, gelatin), as used by this technique, is also described. Experiments in progress have shown that this technique can be used to study the effect of different factors (monoclonal antibodies, lymphokines, etc.) on cell migration, and that it may be used with other cell types which exhibit cell migration on substrates in vitro.

Animals↗

A rapid ELISA for measurement of antibodies to nucleic acid antigens using UV-treated polystyrene microplates.

Pretreatment of polystyrene microplate wells with certain doses of UV light enhances their capacity for binding to single-stranded DNA, double stranded DNA and various synthetic polynucleotides. The use of UV-irradiated plates to immobilize nucleic acid antigens provides a simple, rapid, and specific ELISA for measuring anti-nucleic acid antibodies. The assay is at least as sensitive as the more complex method of precoating plates with poly(L-lysine). It is useful for detection of anti-DNA antibodies in sera of systemic lupus erythematous patients, as well as in culture fluids of murine and human anti-DNA-secreting hybridomas.

Animals↗