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Biomedical subjects

R Ehwald

Publications and source records attributed to R Ehwald.

At least 19 recordsLinked to original sources
PubMed2003

Calibration of the viscometric glucose sensor before its use in physiological liquids--compensation for the colloid-osmotic effect.

The function of the recently described viscometric affinity sensor (VAS), which measures glucose due to its strong effect on the viscosity of a sensitive liquid containing Concanavalin A (ConA) and dextran, was analysed for osmotic and colloid-osmotic effects on the glucose reading. The suction of low- and high-molecular weight osmotica on the membrane of the microdialysis fibre was measured using a membrane osmometer built from the microdialysis probe of the VAS. The reduction of the sensor read-out in blood plasma can be completely explained by a change in small osmotic volume fluxes through the dialysis membrane, which affect the ConA concentration and the viscosity after the flow of the sensitive liquid through the dialysis probe. The measuring error could be prevented by the presence of the polyethylene glycol 6000 at an isotonic concentration in the glucose standard solutions used for sensor calibration.

Biosensing Techniques↗
PubMed2003

Diffusion barriers of tripartite sporopollenin microcapsules prepared from pine pollen.

Tripartite sporopollenin microcapsules prepared from pine pollen (Pinus sylvestris L. and Pinus nigra Arnold) were analysed with respect to the permeability of the different strata of the exine which surround the gametophyte and form the sacci. The sexine at the surface of the sacci is highly permeable for polymer molecules and latex particles with a diameter of up to 200 nm, whereas the nexine covering the gametophyte is impermeable for dextran molecules, with a Stokes' radius > or =4 nm (Dextran T 70), and for the tetravalent anionic dye Evans Blue (Stokes' radius = 1.3 nm). The central capsules obtained by dissolution of the sporoplasts showed strictly membrane-controlled exchange of non-electrolytes, with half-equilibration times in the range of minutes (monosaccharides, oligosaccharides) to hours (dextran molecules with Stokes' radii up to 2.5 nm). The dependence of the permeability coefficients of the nexine for non-electrolytes on Stokes' radius or molecular weight shows that the aqueous pores through the nexine are inhomogeneous with respect to their size, and that most pores are too narrow for free diffusion of sugar molecules. To explain the barrier function of the nexine for Evans Blue, it is assumed that at least the larger pores, which enable slow permeation of dextran molecules, contain negative charges.

Carbohydrate Metabolism↗
PubMed2001

Recording of subcutaneous glucose dynamics by a viscometric affinity sensor.

AIMS/HYPOTHESIS: To provide a nonenzymatic sensor for glucose monitoring in subcutaneous tissue. METHODS: A continuously working affinity sensor based on the glucose-dependent viscosity of a sensitive liquid containing dextran and concanavalin A has been designed by arranging a microdialysis probe, two flow-resisting capillaries and two pressure transducers in a linear flow system. It allows synchronous processing of the viscosity of the sensitive liquid at the standard glucose concentration and the glucose concentration to be measured. In preliminary human trials the sensor was implanted into the subcutaneous tissue of the forearm and its read-out was compared with capillary blood concentrations. RESULTS: In vitro, the viscometric sensor shows a linear and long-term stable dependence on the glucose concentration without detectable drift. At the applied flow rate of the sensitive liquid (about 5 microliters/h) the technical delay is 5 to 10 min. The read-out of the implanted sensor followed the dynamics of the capillary blood glucose concentrations with a time-shift of 10 to 15 min but showed a systematic error when based on precalibration with polymer-free glucose solutions. After appropriate in vivo calibration, the read-out was in good or acceptable coincidence with capillary blood concentrations according to the error grid method and did not show any detectable reduction of sensitivity during the periods of measurement (up to 44 h). CONCLUSION/INTERPRETION: The viscometric-affinity sensor is an efficient tool for current research on glucose monitoring in the subcutaneous tissue and can potentially be further developed for routine clinical use.

Blood Glucose↗
PubMed2000

Additional phosphate stabilises uninterrupted growth of a Dioscorea deltoidea cell culture.

Suspension cells of Dioscorea deltoidea Wall (strain D-1) were maintained in a semicontinuous culture (SCC) in shake flasks at a high growth rate. It was shown that continuous propagation growth of this culture is unstable on Murashige's and Skoog's (MS) medium due to P starvation. On a P-enriched MS-medium the SCC was stable even at mean specific growth rates >0.3 day(-1). Highest volumetric concentrations of furostanol glycosides were obtained, when a P-enriched SCC was not further subcultivated but fed with sucrose. The investigated culture is able to control phosphate uptake and to prevent toxicity on media with excess P. High concentrations of cellular P(i) did not effect the ratio of furostanol to starch.

Journal Article↗
PubMed1999

The Pore Size of Non-Graminaceous Plant Cell Walls Is Rapidly Decreased by Borate Ester Cross-Linking of the Pectic Polysaccharide Rhamnogalacturonan II.

The walls of suspension-cultured Chenopodium album L. cells grown continually for more than 1 year on B-deficient medium contained monomeric rhamnogalacturonan II (mRG-II) but not the borate ester cross-linked RG II dimer (dRG-II-B). The walls of these cells had an increased size limit for dextran permeation, which is a measure of wall pore size. Adding boric acid to growing B-deficient cells resulted in B binding to the wall, the formation of dRG-II-B from mRG-II, and a reduction in wall pore size within 10 min. The wall pore size of denatured B-grown cells was increased by treatment at pH </= 2.0 or by treatment with Ca(2+)-chelating agents. The acid-mediated increase in wall pore size was prevented by boric acid alone at pH 2.0 and by boric acid together with Ca(2+), but not by Na(+) or Mg(2+) ions at pH 1.5. The Ca(2+)-chelator-mediated increase in pore size was partially reduced by boric acid. Our results suggest that B-mediated cross-linking of RG-II in the walls of living plant cells generates a pectin network with a decreased size exclusion limit for polymers. The formation, stability, and possible functions of a borate ester cross-linked pectic network in the primary walls of nongraminaceous plant cells are discussed.

Journal Article↗
PubMed1998

The boron requirement and cell wall properties of growing and stationary suspension-cultured chenopodium album L. cells

Suspension-cultured Chenopodium album L. cells are capable of continuous, long-term growth on a boron-deficient medium. Compared with cultures grown with boron, these cultures contained more enlarged and detached cells, had increased turbidity due to the rupture of a small number of cells, and contained cells with an increased cell wall pore size. These characteristics were reversed by the addition of boric acid (>/=7 &mgr;M) to the boron-deficient cells. C. album cells grown in the presence of 100 &mgr;M boric acid entered the stationary phase when they were not subcultured, and remained viable for at least 3 weeks. The transition from the growth phase to the stationary phase was accompanied by a decrease in the wall pore size. Cells grown without boric acid or with 7 &mgr;M boric acid were not able to reduce their wall pore size at the transition to the stationary phase. These cells could not be kept viable in the stationary phase, because they continued to expand and died as a result of wall rupture. The addition of 100 &mgr;M boric acid prevented wall rupture and the wall pore size was reduced to normal values. We conclude that boron is required to maintain the normal pore structure of the wall matrix and to mechanically stabilize the wall at growth termination.

Journal Article↗
PubMed1996

Viscosimetric affinity assay.

Affinity ligands and/or affinity receptors may be quantified by a viscosimetric assay which can be carried out with a simple technique and has the potential of broad applications. The viscosimetric affinity assay is based on the high contribution of affinity bonds to the viscosity of an aqueous dispersion of a hydrocolloid that is bearing affinity ligands. In dispersions of such sensitive hydrocolloids at a concentration above the overlapping point, agglutination is not possible and the modulation of viscosity by the formation or dissociation of intercolloidal affinity bonds may be several orders of magnitude larger than the basic viscosity measurable in the absence of intercolloidal affinity bonds. If dispersions (30 g liter-1) of branched dextran with high molecular weight were used as reagent for concanavalin A (Con A), the Con A concentration necessary for a significant rise in viscosity was decreased with increasing colloid size. The viscosity of dispersions containing both a ligand-bearing high-molecular-weight dextran and an appropriate polyvalent receptor protein (lectin or antibody) showed a dependence on the concentration of free ligands (sugars or insulin) according to the law of mass action. In this competitive mode the viscosimetric affinity assay seems to be well adaptable to many analytical problems.

Antibodies↗
PubMed1993

Entrapment of dextran in plant cell capsules by reversible change of cell wall permeability.

Vesicular packing material (VP) made of clusters of extracted higher plant cells with the intact framework of their cell wall was used so far for permeation chromatography (vesicle chromatography). The objective of this study was to devise a method to entrap dextran in the vesicles. This can provide a means to entrap biocatalysts and secondly, to create aqueous two-phase systems with a stationary dextran phase for liquid-liquid partition chromatography. Dextran of molecular sizes above the separation limit of the plant cell wall cannot permeate into the intracellular space in aqueous medium. However, in hydrophilic organic solvent/water mixtures, dextran molecules can diffuse into the capsules. The removal of the organic solvent leaves the dextran trapped inside. There was an inverse correlation between the percentage of dextran permeating through the cell wall (Pperm) and the concentration of solvent required for dextran precipitation. The increase of permeability is therefore considered to be caused, to a great extent, by the decrease of the effective size of dextran molecules due to decreased solvation. Pperm was inversely correlated to the dielectric constants and the polarities of the solvents and, in the case of protic solvents, the hydrogen-bond acidities. No correlation was found to the hydrogen-bond basicities.

Cell Membrane Permeability↗
PubMed1992

Inclusion and fractionated release of nucleic acids using microcapsules made from plant cells.

The encapsulation and fractionated release of nucleic acids on vesicular packing (VP) materials have been investigated. The earlier described dependence of the permeation of nucleic acid molecules through the vesicle membranes on the salt concentration is a necessary precondition for both encapsulation and fractionation. Encapsulation is achieved by applying a suitable sample onto a VP column that has been equilibrated with a high-salt buffer. In that buffer the sample molecules are permeable. Immediately after sample application, elution is started with a low-salt buffer, from which the sample molecules are excluded. At the front between the two buffers the permeability changes, and some of the sample molecules distributed inside the vesicles cannot pass through the membranes. These encapsulated molecules can be released by increasing the salt concentration in the eluent. If the encapsulated nucleic acid sample is polydisperse, a stepwise or linear increase in the salt concentration leads to a fractionated release. The fractions obtained differ in their molecular size composition.

Base Sequence↗
PubMed1992

[Permselect--a new substance for leukocyte elimination].

Permselect particles consisting of purified plant cell wall ghosts obtained from the biomass of a suspension culture of Chenopodium album L. show a very high efficiency for leukocyte binding. A package containing 1 g dry Permselect eliminates 99-100% of leukocytes from 80 ml of fresh blood, preserved whole blood, or red blood cell concentrates. At comparable conditions Permselect shows a higher efficiency of leukocyte binding than commercial filter material (cotton, cellulose acetate). The vitality parameter of filtered erythrocytes and their stability during storage were not altered. The data are discussed with respect to causality of leukocyte binding to the pectin-containing surface of the cell wall particles.

Adsorption↗
PubMed1990

[Exclusion chromatography for the separation of cryoprotective agents from freeze-preserved blood cells].

A method for separating low molecular cryoprotectiva from freeze-conserved erythrocyte- and thrombocyte-concentrates by exclusion chromatography has been described. A new vesicular packing material has been used. Only 25 to 30 minutes are necessary in order to separate glycerol respectively dimethylsulphoxide (DMSO) completely from the cells. 86.5% of the erythrocytes and 75.4% of the thrombocytes were recovered after the separation process.

Blood Platelets↗
PubMed1990

[Hemapheresis using vesicular plant separation materials].

The present paper deals with the separation of cells from soluble compounds of blood by means of exclusion chromatography using a recently described vesicular packing material made from the cell wall framework of the small duckweed Wolffia arrhiza. The cells of the periphere blood are hardly retarded in passing through a packing of the vesicular material and eluted as sharp peak at an elution volume which is near to 30% of the column volume. The behavior of cells is similar to that of the excluded high molecular weight plasma proteins (e.g. serumalbumin). Low molecular weight solutes (e.g. salts, glucose, urea, kreatinin), but also substances of considerable molecular weight (e.g. myoglobin and Vitamin B12) which are usually difficult to separate by dialysis from serum, are eluted at nearly 100% of the packing volume and may be separated completely from cells and high molecular weight proteins. In vitro-Tests did not show a reduced vitality of eluted blood cells.

Blood Component Removal↗
PubMed1981

Active transport and mediated diffusion of glucose and other monosaccharides in Endomyces magnusii.

After growth on sucrose or glucose, Endomyces magnusii possess a monosaccharide uptake which resembles that of Saccharomyces cerevisiae (a high KT of uptake, preference for alpha-anomers of D-xylose and D-glucose, enhanced uptake during anaerobiosis, attainment of a diffusion equilibrium). The uptake is inhibited by other monosaccharides and especially strongly by D-galactose. In the absence of high concentrations of metabolizable sugars. E. magnusii develops a capacity to accumulate 3-O-methyl-D-glucose and D-xylose against a concentration gradient the new system displaying a high affinity for glucose (KT less than 0.1 mM), repression by glucose, mannose or galactose. Cycloheximide (0.2%) blocks the formation of the active system.

Ascomycota↗
PubMed1981

Anomeric specificity of the monosaccharide carrier in yeasts and yeast-like organisms.

The anomeric specificity of monosaccharide uptake was investigated in 42 species of yeasts and related mycelium-forming fungi. Differences in the uptake of anomers were determined by the following methods. (1) Shift of anomeric equilibrium in the outer medium caused by preferential uptake of one of the anomeric forms was monitored polarimetrically as induced mutarotation. (2) The uptake of 14C-D-glucose by cells was examined after addition of freshly prepared solutions of alpha- or beta-D-glucose. Most of the organisms examined display the Saccharomyces-type preference for the alpha-anomers of glucose and xylose which is caused by the higher affinity of the monosaccharide carrier for the alpha-pyranose configuration. The following genera show this type of preference (the number of species is given in parenthesis): Saccharomyces (5), Schizosaccharomyces (1), Endomycopsis (2), Eremascus (1), Endomyces (1), Pichia (1), Hansenula (1), Debaryomyces (2), Lipomyces (1), Willia (1), Nematospora (1), Kluyveromyces (2), Candida (5), Torulopsis (5), Cryptococcus (1). No anomeric specificity was shown by the following genera: Nadsonia (1), Dipodascus (2), Rhodotorula (5), Sporobolomyces (2), Bullera (1), Rhodosporidium (1). A parallel investigation of the concentration dependence of glucose uptake indicates that most yeasts possess a constitutive monosaccharide carrier characterized by the following features: a high maximum rate of uptake, a relatively low affinity, and preference for alpha-anomers. Besides this carrier the majority of these microorganisms possess a glucose-transporting carrier with a higher affinity and a lower capacity.

Biological Transport, Active↗
PubMedDate not supplied

Compensation of temperature and concanavalin A concentratration effects for glucose determination by the viscometric affinity assay.

A viscometer suitable for rapid measurements in small volumes of highly viscous liquids is described. Using this device the viscometric affinity assay for glucose was studied under variable conditions in order to obtain basic information for the design of a viscometric glucose sensor. The viscosity of the dextran/Concanavalin A (ConA) solution is sensitive to glucose in a broad range of the shear stress. However, for measuring the glucose concentration with this sensitive liquid the strong dependence of the absolute viscosity on temperature and ConA concentration has to be taken into account. For the purpose of calibration a parameter more suitable than the absolute viscosity is the relative fluidity (F(r)) that is defined by the actual measured viscosity at a given glucose concentration, the reference viscosity at a standard glucose concentration, and a constant linearization coefficient. F(r) shows a linear dependence on the glucose concentration in the therapeutically interesting range up to 30 mM and is not significantly changed by moderate variations of the ConA concentration or temperature.

Concanavalin A↗