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Biomedical subjects

M Bier

Publications and source records attributed to M Bier.

At least 37 records · Page 2Linked to original sources
PubMed1993

A new buffering system and its use in electrophoresis and isoelectric focusing.

A novel buffer system with rather unique properties particularly well suited for preparative isoelectric focusing is described. The system was developed in response to the long standing need for pH gradients formulated using inexpensive components of known chemical composition. The system is binary consisting of buffer pairs covering the pH range of 3 to 10, in increments of less than 1 pH. The components are all readily available, nontoxic, and biologically acceptable, and are selected on the basis of their dissociation constants. The requirement for each buffer pair is that there be an overlapping pH zone where both components are largely non-ionized, having a ratio of neutral species to total buffer concentration between about 0.90 and 0.99. This ratio is defined as the 'electrophoretic reserve capacity' (ERC). Within the above ERC range a stable pH gradient is obtained, assuring sharply resolved protein focusing. Buffer parameters, mainly pH, buffering capacity, ERC, transport numbers, and conductivity, can be easily calculated and the buffer composition adjusted to meet requirements. All components are used at high concentration, typically of the order of up to 100 mM, yielding buffering capacities and conductivities analog to those customary in electrophoresis.

Acids↗
PubMed1993

Counterflow in isotachophoresis: computer simulation and experimental studies.

A computer model has been developed to predict the effects of co- and counter-current flows on electrophoretic transport. Model predictions are compared with experimental data obtained with the large-scale recycling isotachophoresis apparatus, recently developed in our laboratory. There were no significant changes in the steady state properties or the transient development of the Kohlrausch-adjusted profiles with and without flow. The application of counterflow in free flow isotachophoresis in early stages, before a steady state is achieved, can result in selective wash-out of slower sample components. Thus, the magnitude of counterflow can be used as a variable in optimizing separations. The characteristic features of the model render it also suitable for the simulation of electroosmotic flow in capillary electrophoresis in open tubes, where plug-type flow prevails.

Anions↗
PubMed1992

Recycling isoelectric focusing: use of simple buffers.

Using the recycling free-flow focusing (RF3) apparatus, we have demonstrated that single ampholytes can be utilized to establish very stable pH regions, separating all proteins into three groups: a sharply resolved zone of proteins isoelectric at the prevailing pH, this "pH window" being bracketed by zones of more acidic and/or basic proteins. The ampholytes used are either amino acids or their dipeptides and other derivatives. Where necessary, because of lack of an ampholyte with the required pH, a binary mixture of ampholytes can be utilized. The closer their isoelectric points (pI), the narrower will be the pH window, i.e., the sharper the resolution of the bracketed proteins. This method overcomes the necessity of using ill-defined commercial carrier ampholytes, such as Ampholine, for preparative isoelectric focusing. It is recommended that the ampholytes be utilized at relatively high concentration, 100mM or higher, this contributing to pH stability and minimizing protein precipitation.

Amino Acids↗
PubMed1990

Electrophoretic transport of solutes in aqueous two-phase systems.

Electrophoretic transport of proteins across the interface between the phases of an aqueous polymer two-phase system can be greatly impeded in comparison with transport within the individual phase. This effect can be controlled by modifying the affinity of the protein for a phase by suitable manipulations of such variables as pH. The effect is not caused by differences in the electrophoretic velocity between the two phases, nor by large changes in pH at the interface. An analogy exists between this phenomenon and the related subject of diffusion of electrolytes across the phase interface.

Buffers↗
PubMed1990

Membrane active plasma factor in multiple sclerosis: characterization and isolation by recycling isoelectric focusing.

Recycling isoelectric focusing is a rapid, high resolution technique that has the capability of fractionating complex mixtures of proteins on a preparative scale on the basis of their isoelectric points (pIs). For this reason, it appeared to be an ideal tool to further characterize and isolate the surface active plasma component(s) which is abnormal in multiple sclerosis (MS). The normal control and the abnormal MS plasma components, or factors, proved to be stable under the conditions used in this technique, including deionization by electrodialysis, dialysis against distilled water, lyophilization and the presence of 3M urea and carrier distilled water, lyophilization and the presence of 3M urea and carrier ampholytes. The presence or absence of plasma factor activity was determined by incubating red blood cells in a test plasma, or plasma fraction, followed by the determination of the red blood cell electrophoretic mobility in the presence and absence of linoleic acid. Both the normal and MS factor had a pI of 4.0 +/- 0.1 under the conditions used. A high degree of purification was achieved and albumin was eliminated as a possible candidate for the factor(s).

Dialysis↗
PubMed1989

Computer simulation and experimental validation of the electrophoretic behavior of proteins.

A mathematical model of the electrophoretic behavior of proteins is presented. The Debye-Hückel-Henry theory is used for the description of protein mobility, which has the important result of making net mobility a function of ionic strength. A net charge vs pH relationship and a diffusion coefficient are required to describe a specific protein. The model is employed for the computer simulation of three distinct electrophoretic modes: isoelectric focusing, isotachophoresis, and zone electrophoresis. The validity of the model is tested by comparing simulation with experimental data. Excellent qualitative agreement was found.

Computer Simulation↗
PubMed1988

Purification of lentil lectins using preparative electrophoresis.

Electrophoresis presents an interesting alternative to chromatography for the purification of biological compounds. To document the performance of three preparative electrophoresis apparatus currently available, they were applied to the purification of lectins from lentil seeds which contain two isolectins usually purified by chromatography. Purification by electrophoresis consists of first isolating a mixture of the two isolectins and then separating them. For the first step, either of two free-flow electrophoresis apparatus were employed: the Elphor VaP 22, using field step electrophoresis and the Biostream using zone electrophoresis. To optimize the process, the Biostream was modified to a recycling mode. This required repositioning one dialysis membrane which separates an electrode from the separation chamber. This allowed the lentil extract to be desalted by electrodialysis directly in the apparatus prior to fractionation. A high concentration of lectins was collected at the cathode and acidic proteins were collected at the anode. The bulk of the extract was recycled until the whole volume was processed. In a second step the isolectins were separated by recycling isoelectric focusing in the recycling isoelectric focusing apparatus. The present work clearly demonstrates that electrophoretic methods provide lectins with higher purity than chromatographically purified commercial products.

Electrophoresis↗
PubMed1987

Low-level fetal lead exposure effect on neurobehavioral development in early infancy.

A prospective method was used in this study to assess the effects of fetal lead exposure on neurodevelopmental status in 3- and 6-month old infants. At their first prenatal medical appointments, 305 lower socioeconomic status women residing in predesignated lead-hazardous areas of Cincinnati were recruited. Lead was measured in whole blood in both the mother and fetal-placental unit (prenatal and cord) and the neonate (ten days and 3 months). All blood lead levels were less than 30 micrograms/dL. Infant development was assessed with the Bayley scales at 3 and 6 months of age. Multiple regression analyses which treated perinatal health factors such as birth weight and gestation as confounders indicated an independent, inverse relationship between both prenatal and neonatal blood lead levels and performance on the Bayley Mental Developmental Index at both ages. Male infants and infants from the poorest families appeared to be especially sensitive to these psychoteratogenic influences. Further study using a structural equations approach indicated that neurobehavioral deficits were partly mediated by lead-related reductions in birth weight and gestation.

Age Factors↗
PubMed1985

Characterization of synthetic carrier ampholytes by repetitive scanning of the electric field during focusing.

This paper reports the utilization of a potential gradient array detector for monitoring the dynamics of the electric field during isoelectric focusing. Transient and steady state electric field profiles are presented for synthetic carrier ampholyte mixtures with a wide (approximately 3-10) pH range. Two available commercial products (Ampholine and Pharmalyte) and a laboratory synthesized mixture (PEHA ampholytes) are compared. The formation of conductivity gaps and their migration toward the cathode in extended experiments (cathodic drift) can be visualized with this system.

Ampholyte Mixtures↗
PubMed1983

Electrophoresis: mathematical modeling and computer simulation.

A mathematical model of electrophoretic separation processes has been developed and adapted for computer simulations. The model is used to predict the characteristic behavior of a variety of electrophoretic techniques from a knowledge of chemical equilibria and physical transport phenomena. The model provides a unifying basis for a rational classification of all electrophoretic processes.

Computers↗
PubMed1983

Properties of narrow-range ampholytes isolated from wide-range ampholyte preparations.

A simple procedure for obtaining useful narrow-pH-range ampholytes from inexpensive laboratory-synthesized ampholytes by preparative isoelectric focusing in Pevikon is described. The narrow-range ampholytes prepared in this way are comparable to commercial ampholyte preparations as judged by conductivity, buffer capacity, pH gradient formation, and resolving power. These inexpensive narrow-range ampholytes are particularly well suited to preparative isoelectric focusing applications requiring large quantities of ampholytes.

Ampholyte Mixtures↗
PubMed1976

Serum protein fractionation by isotachophoresis using amino acid spacers.

Analytical and preparative isotachophoresis has been carried out using amino acids and peptides as discrete spacers in contrast to the usually employed continuous mobility spectrum Ampholine. Analytical isotachoresis in free solution, using the LKB Tachophor, demonstrated the separation of human serum into distinct mobility subgroups, n spacers giving rise to n + 1 protein subgroups. Preparative fractionation on polyacrylamide gel was carried out on the LKB Uniphor using threonine and glycine as spacers. Immunoelectrophoretic analysis showed that eight out of ten proteins assayed were clearly resolved in the three subgroups obtained, thus demonstrating the sharpness of isotachophoretic resolution.

Amino Acids↗
PubMed1975

Hypochlorite-induced alterations to canine serum complement.

Changes in the concentration of the components of complement produced by NaOC1 both in vitro and in vivo are recorded. C1, C4 and C7 are particularly sensitive to this oxidizing agent, although all components decrease at high concentrations of NaOC1. Following oxidation, complement componenets return rapidly to normal. Data are presented to indicate that part of this repair mechanism is due to the action of reducing agents such as ascorbic acid and part is due to the synthesis of the individual components. The unique sensitivity of complement components to oxidation make this treatment of potential value in suppressing the inflammatory response.

Animals↗