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Biomedical subjects

G Ruhenstroth-Bauer

Publications and source records attributed to G Ruhenstroth-Bauer.

At least 37 records · Page 2Linked to original sources

Biological and clinical relevance of human macrophage migration inhibitory factor (MIF).

The first isolation and characterization of a lymphokine, the human MIF (MSF), and the availability of a specific MIF (MSF)-antibody provide an opportunity to establish a new parameter for detection of cellular immune mechanisms in various clinical situations. The precise estimation of quantitative amounts of MIF by sensitive immunochemical techniques could gain new insights into the pathogenesis of a number of clinical disorders. In addition, the exact detection of quantitative amounts of MIF in body fluids could have prognostic value.

Antibodies

Simultaneous flow cytometric DNA and volume measurements of bone marrow cells as sensitive indicator of abnormal proliferation patterns in rat leukemias.

Simultaneous flow cytometric DNA and volume analysis of normal rat bone marrow cells shows three populations of nucleated cells with different mean volume. Each of these populations proliferates in a distinct cell cycle (alpha, beta, gamma). Normally the alpha-cell cycle has the highest amplitude, the beta-cell cycle is intermediate, and the gamma-cell cycle is low. The alpha-cell cycle was very significantly depressed and the beta + gamma-cell cycle was increased in three different rat leukemias (L5222, Shay, BNML), growing on three different rat strains (BDIX, Holtzmann, Brown Norway). The two parameter analysis further revealed that cells of the beta + gamma-cell cycle were slightly hyperdiploid and hypertetraploid in leukemic animals. The decrease of the alpha-cell cycle and the hyperploidies were more sensitive indicators for the abnormal proliferation pattern than the analysis of one parameter DNA distributions which remained within normal limits in all three leukemias.

Animals

Flow cytometry as a new method for the measurement of electrophoretic mobility of erythrocytes using membrane charge staining by fluoresceinated polycations.

The binding of FITC-labeled poly-L-ornithine and poly-L-lysine to fresh or neuraminidase treated human, rat or rabbit erythrocytes was investigated by simultaneous cell volume and cell membrane fluorescence measurements in a flow cytometer. The cell volume was converted into cell surface and the distribution curve of the fluorescence/micrometer2 cell surface was calculated from all histogram classes by a computer program. The mean fluorescence/micrometer2 cell surface as a measure of the density of the negative charges on the cell surface was directly proportional to the elctrophoretic mobility of the erythrocytes, showing that polycation binding can effectively be used for the measurement of the electrophoretic mobility of erythrocytes. The computer fitting of the experimental two parameter histograms by two dimensional Gaussian normal distributions was found to be a very efficient way of data reduction, and a good separation of overlapping cell clusters was possible even in the case of low total numbers of cells in the histogram.

Animals

[The specific effect of plasma proteins in erythrocyte sedimentation. An analysis of the correlation coefficients between ESR and the concentration of twenty individual plasma proteins in healthy and sick persons, with special reference to myocardial infarction (author's transl)].

1. Spearman's rank correlation coefficient between erythrocyte sedimentation rate and the concentration of twenty individual plasma proteins was determined in 72 persons including 37 patients with recent myocardial infarction. The highest correlation coefficients with ESR could be demonstrated in the following proteins: fibrinogen, alpha-1-acid-glycoprotein, alpha-2-macroglobulin, alpha-1-antitrypsin, coeruloplasmin, Ig M. The closest correlation with ESR was found, when the molar concentrations of fibrinogen, alpha-2-macroglobulin and Ig M were summed up. 2. Subdivision of the examined group of patients according to their diagnosis showed that the degree of correlation with ESR in many plasma proteins essentially depends on the composition of the studied group of patients. The patients with myocardial infarction, with pneumonia and those with neoplasma all showed distinctly different patterns of correlation. These obviously reflect underlying changes in the concentrations of the studied proteins, that seem to be specific to the respective disease. 3. In 4 patients with myocardial infarction the changes in plasma protein concentrations together with ESR were followed over a period of 26 days after the infarction.

Blood Proteins

Human migration inhibitory factor: purification and immunochemical characterization.

Using gel filtration and preparative isotachophoresis, the migration inhibitory factor (MIF) was highly purified from human lymphocytes activated with concanavalin A. MIF is an acidic protein with a mol wt of approximately equal to 25,000 daltons as determined by gel filtration and analytical polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The protein inhibits migration of macrophages in the capillary test and in addition, has a slowing effect on the electrophoretic mobility of guinea pig peritoneal macrophages. Rabbit antibodies specific for this protein, as determined by immunochemical techniques, neutralized the biological effect of MIF on migration and on the electrophoretic mobility of macrophages.

Antigen-Antibody Reactions

[Concentration and characterization of a liver cell proliferation factor from partially hepatectomized rat livers (author's transl)].

An extract of rat liver, made 12 h after partial hepatectomy, was injected into untreated rats. The synthesis of DNA in the liver cells of the injected animals showed a 3.3 fold increase in comparison with controls. A similar increase was observed in the mitosis index. Injection of liver extracts from normal rats showed no effect when compared with the injection of NaCl. The effect of the active extract was not altered by neuraminidase treatment. Trypsin-chymotrypsin treatment, howerver, removes the activity. The active factor must therefore be either a protein or a protein-like substance. The molecular weight of the factor was shown to be between about 30 000 and 50 000 by molecular filtration. The specific activity after molecular filtration was raised by a factor of 100. The factor is inert with respect to the proliferation of kidney and spleen. It is apparently not species-specific, however, since it also causes increased proliferation of liver cells in normal mice.

Animals

The trajectories of particles suspended in electrolytes under the influence of crossed electric and magnetic fields. Possible explanation of the sensitivity of organism to magnetic fields.

We observed that particles, suspended in an electrolyte and brought into crossed magnetic and electric fields of low intensities, will deviate in the central part of the electrophoresis chamber of a standard Zeiss Cytopherometer with a component vertical to both fields. The direction and magnitude, however, were sharply at variance with what would be expected by the action of the Lorentz force (EMF) on the surface of the particles. The magnitude of the deviation depends upon the magnetic and electric field strength, the ion concentration of the suspension medium and the geometry of the chamber. The movement of the particles is due to streaming of the electrolyte which is mainly caused by inhomogeneities of the electric field in the electrophoresis chamber. The magnitude of the effect is high enough to occur physiological conditions. Magneto-electrophoretic streaming might eventually act as a transducer mechanism which could explain the ability of some animals to orientate themselves in the geomagnetic field.

Electricity

[The regulation of acute inflammatory processes by systemic changes of plasma protein profiles].

Local inflammatory processes in the human as well as in experimental animals cause a selective increase or decrease of the hepatic synthesis rate of many plasma proteins. The resulting systemic changes in the plasma-protein profile regulate the extent of the local inflammatory response. The physiologic importance of this feedback mechanism is directed at preventing the spread of local inflammatory tissue destruction. By means of mediator substances involved in this regulatory system, the infalmmatory responses can be experimentally manipulated.

Animals

Fluvo-metricell, a combined cell volume and cell fluorescence analyzer.

A new flow through instrument that simultaneously measures cell volume (resistance pulse technique) and cell fluorescence in the same orifice will be described. The fluorescence pulses of the hydrodynamically focussed cells are picked up by the optics via the axial direction (principle of Dittrich and Goehde, Z Naturforsch 24b:360, 1969). There is no coordination problem between the fluorescence and the resistance pulses to be observed because a new type of transducer is used. The electronic system provides gating of one or two parameter histograms. Function tests are performed with the incorporated two-parameter test spectrum generator. Different examples of using the instrument in practice are shown. The volume that may be measured with an orifice of 70 micron diameter ranges between 4 and 1400 micron3 (1:350). Coefficients of variation of the fluorescence below 2% are measured.

Cytological Techniques

A volume-activated cell sorter.

A sorter that is activated by the resistance pulses of a Coulter orifice (volume detector) has been developed. The problem arising from the distortion of the volume signals produced by the electric droplet forming and charging voltages are avoided by special shielding and grounding of the volume detector. Experiments with beads and leukocytes are described. The maximal processing rate at this time is 2000 cells/sec, the purity of sorted fractions is better than 97%, and the viability of sorted cells is better than 80%.

Cell Separation

Erythrocyte sedimentation rate and C-reactive protein.

C-reactive protein (CRP) was isolated from pleura exudate of patients with metastasing cancer and added to plasma of healthy blood donors. Only the addition of excessive amounts of CRP (220 mg/ml) caused a notable increase in erythrocyte sedimentation rate. An immunoelectrophoretic method for the simultaneous quantitative determination of CRP together with C 3 or other acute phase reactants is described.

Blood Sedimentation

Plasma protein profiling: the diagnostic evaluation of disorders in plasma protein composition by a new immunoelectrophoretic method.

A new electroimmunoprecipitation technique is presented by means of which a great variety of antigens e.g. plasma proteins can be simultaneously and quantitatively determined with a single-step electrophoretic separation. The essential features of the new technique are: (a) subdivision of the antibody gel into gel strips containing monospecific antibodies to individual plasma proteins. (b) sample application as a "sample gel" filling a trough over the width of the immunoplate. Quantitation is based on the fact that the distance an antigen can migrate within a gel containing a defined amount of specific antibody directed against the antigen is determined by the concentration of the appropriate antigen within the sample. The area where the antigen is finally completely consumed by immunoprecipitation and antibody present in excess is sharply delineated. The applicability of the method in simultaneous quantitative determination of 15 plasma proteins is demonstrated with plasma from healthy blood donors and patients with various diseases. The advantage of the new technique as compared to commonly used clinical acetate folia electrophoresis is the high degree of specificity for the determination of a great number of individual, diagnostically meaningful plasma proteins. The advantage over common quantitative two-dimensional immunoelectrophoresis is its uncomplicated way of evaluation. The potential clinical application of the new quantitative immunoelectrophoretic technique in diagnostic screening and differential diagnosis is discussed.

Blood Proteins

The significance of plasma lipoproteins on erythrocyte aggregation and sedimentation.

Increased erythrocyte aggregation can be induced by high concentrations of human lipoproteins. The dependence of aggregate formation on lipoprotein concentration was recorded by determination of erythrocyte sedimentation rate (ESR), by electrical measurement of the erythrocyte aggregation index (EAI) and by scanning electron microscopy. The lipoprotein concentrations necessary to induce a significantly increased ESR in an otherwise normal human plasma are much too high to be encountered in physiologic or even severe pathologic states. Therefore hyperlipoproteinaemia by itself cannot explain a raised ESR. In cases where the ESR is raised due to the presence of increased amounts of other erythrocyte aggregating plasma proteins (agglomerins), hyperlipoproteinaemia can contribute to a limited extent to the increase in ESR. The possible pathophysiological significance of the demonstrated erythrocyte aggregating capacity of human lipoproteins in a microvascular environment is noted.

Blood Sedimentation