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Biomedical subjects

M Wagner

Publications and source records attributed to M Wagner.

At least 1,099 records · Page 61Linked to original sources

Bacterial decontamination and antileukemic therapy of AKR mice.

Four nonabsorbable antibiotics (streptomycin, neomycin, bacitracin, and amphotericin B) and a germicidal dip solution (Zephiran chloride/water) were used to eliminate all the detectable bacteria from conventional AKR mice. Control mice were not decontaminated and were used as such. When antibiotic-decontaminated and control mice developed clinical manifestations of spontaneous lymphatic leukemia, each was treated for the disease with an antitumor drug (cyclophosphamide [CP]) at weekly intervals. With the decontamination procedure, mice of each of the two groups became bacteria-free after 16 weeks of continuous oral administration of the antibiotics and two separate germicidal dippings. All decontaminated mice remained free of bacteria throughout the experiment. The bacterial flora of the control mice remained unaltered. With CP therapy, the mean survival time of the female decontaminated mice was 65 days, whereas that of male mice was 218 days. The average survival time of the CP-treated control leukemic mice was 51 days. Untreated decontaminated or control mice usually died of leukemia within 7 days after the onset of symptoms of leukemia. Although CP therapy was not curative, it did prolong the life expectancy of the decontaminated mice significantly.

Amphotericin B↗

The size pH, and redox potential of the cecum in mice associated with various microbial floras.

Cecal size and in situ redox potential and pH of cecal contents were determined in conventionally reared mice and mice reared under a variety of gnotobiotic conditions: germfree, monoassociated with a cecal Clostridium sp., hexaflora-associated and thermoduric polyflora-associated. The mean Eh was approximately +200 mV in germfree and -200 mV in conventional mice. The Eh was close to zero in the monoassociated mice, thus occupying a position intermediate between the germfree and conventional mice. The potentials observed in the hexaflora and the thermoduric flora groups were indistinguishable from those of conventional animals. The degree of normalization was more advanced with respect to the redox potential than to the cecal size in the various gnotobiotic groups. In the thermoduric polyflora-associated group, normalization was observed in both cecal size and redox potential. This demonstrates that normalization can be accomplished with a relatively simplified microflora, at least with regard to the parameters studied.

Animals↗

Effect of Corynebacterium parvum on the response to irradiation of a C3H fibrosarcoma.

A methylcholanthrene-induced fibrosarcoma of C3H mice has been used as 5th- to 6th-generation syngeneic transplant in an investigation of the role of Corynebacterium parvum as an adjuvant to the therapeutic effect of local irradiation of established tumors. The most effective route for administration of C. parvum in this tumor system was i.v., and the greatest effect of the C. parvum-mediated tumor graft rejection was observed for tumor growing intracutaneously or s.c. An intermediate level of effectiveness was obtained for tumor growing i.m. The combined C. parvum and local irradiation studies were performed using tumors growing in the leg muscle and measuring 8 mm in diameter at the time of local irradiation. Several routes of administration of C. parvum, dose levels of C. parvum, and time relationships between administration of C. parvum and irradiation were investigated. The outstanding finding was that very low doses of radiation were quite effective when administered to tumors growing in animals pretreated with C. parvum. This was true for radiation administered as a single or fractionated dose (10 equal doses spread over 18 days). For single-dose irradiation the effect was relatively dose independent over the range of 200 to 3000 rads. Some indication was obtained that local irradiation may impair the tumor graft rejection reaction. No evidence was obtained of an enhanced growth of tumor. Regression of tumor following irradiation was not modified by pretreatment with C. parvum. The mice that were unsuccessfully treated by radiation and C. parvum had a lower incidence of metastatic disease in the lung than did the mice treated unsuccessfully with radiation alone.

Adjuvants, Immunologic↗

[Electron microscopic detection of blood group antigen A on human erythrocytes by means of ferritin- and gold-labelled protectin of helix pomatia].

By means of ferritin- and gold-labelled protectin from the albumen gland of the edible snail Helix pomatia the blood group antigen A was located on human erythrocytes of groups A1, A2 and A1B. With erythrocytes of groups O and B the reaction is negative. The antigen is focally distributed on the outer surface of the cell membrane. Cells from groups A1 and A1B have an antigen A concentration about 4 times greater than A2 cells. The numbers of particles after tagging with ferritin or gold are comparable. The gold particles show an extremely high contrast and are therefore very suitable for the immunoelectron microscopic localization of antigens.

ABO Blood-Group System↗

Immunoglobulins under the influence of nonspecific factors. III. Immunoglobulins, pertussis antibodies and lysozyme in three child populations exposed to different air pollution.

The levels of IgG, IgA, IgM, lysozyme, agglutinins against B. pertussis and B. parapertussis were followed in the blood serum of 306 children 9--10 years old in 3 areas of Central Bohemian region. In children dwelling in areas with more polluted air significantly higher levels of serum lysozyme and of parapertussis antibodies were found by the t-test. The distribution of these values shows significant differences between more polluted areas in comparison with lower-pollution area also in Kolmogorov-Smirnov test. The average levels of Ig approached statistically critical values but did not reach them, but significant differences in the distribution of values of IgG and IgA were shown by the F-test and chi2-test between lightly and heavily polluted areas. The values of immunological reactions in polluted areas were always higher than in the non-polluted group. This provides evidence for a hypothesis about a stimulatory effect of polluted air on immunological mechanisms in child population. The higher values of IgM observed recently by other authors in women were shown in girls of 9--10 years.

Agglutinins↗

Electron microscopic demonstration of cell surface carbohydrates by means of peroxidase and ferritin complexes of the Lens culinaris lection.

The use of Lens culinaris lectin for electron microscopic detection of D-mannose,- D-glucose and N-acetyl-D-glucosamine like sites on tumor cells, erythrocytes, erythrocyte ghosts, cultured rat liver cells and various tissues of mice is demonstrated. In addition to Lens culinaris lectin-peroxidase reaction (LeL-po reaction) the preparation of active Lens culinaris lectin-ferritin conjugate are described and the specificity of cytochemical reactions are demonstrated. Furthermore experiments by immuno freeze-etching are reported for topological analysis of the lectin receptors.

Acetylglucosamine↗

[Studies on the Concanavalin A-receptors of rabbit erythrocytes and of erythyrocyte shadows using immunoelectron microscopy].

The authors report on the immuno electron microscopic demonstration of Concanavalin A-receptors on the erythrocyte membrane. Immuno freeze-etching studies evidenced a focal arrangement of Concanavalin A-receptors on intact erythrocytes as well as on erythrocyte ghosts. Obviously hemolysis by distilled water does not produce changes in the pattern of distribution of Concanavalin A-receptors whereas slight aggregation of plasma membrane intramembraneous particles occured. No unambigous spatial relationship between Concanavalin A-receptors and the plasma membrane intramembraneous particles (intrinsic proteins) of erythrocytes could be observed.

Animals↗

Wheat germ agglutinin and Ticinus communis agglutinin as specific saccharide stains in light and electron microscopy.

Two plant agglutinins, wheat germ agglutinin and Ricinus communis agglutinin, were used for light and electron microscopic detection of certain carbohydrate-containing cell surface components and extracellular polysaccharides. For light microscopic studies on various tissues fluorescein isothiocyanate coupled lectins were prepared. The ferritin coupling of the lectins for electron microscopy was performed by glutarldehyde in the presence of the specific hapten. The specificity of the reactions was demonstrated by blocking with the hapten.

Antibody Formation↗

Possible peptide chain termination mutants in thymide kinase gene of a mammalian virus, herpes simplex virus.

Mutations in the viral gene coding for the thymidine kinase (ATP:thymidine 5'-phosphotransferase, EC 2.7.1.75) induced by herpes simplex virus have been obtained by selection of virus resistant to bromodeoxyuridine when grown in thymidine-kinase-deficient LMTK- mouse cells. Proteins labeled after infection of Vero (monkey) cells with herpes simplex virus were analyzed by gel electrophoresis and one protein of about 40,000 daltons was consistently altered in a number of thymidine-kinase-deficient mutants. Many viral mutants lacked this peptide and one class of these mutants induced the synthesis of new shorter peptides. Revertant virus could be selected which simultaneously regained the ability to induce thymidine kinase activity, regained the intact thymidine kinase peptide, and lost the ability to synthesize the shorter peptide fragment. These mutants comprise a class of animal virus mutants which have the properties expected of peptide chain termination mutants.

Bromodeoxyuridine↗

Decontamination of gnotobiotic mice experimentally monoassociated with Candida albicans.

Gnotobiotic AKR mice, experimentally monoassociated with Candida albicans, were successfully decontaminated by oral treatment with amphotericin B incorporated in the drinking water. Germfree mice first were swabbed orally with viable C. albicans and then were allowed to acclimatize for 4 weeks. The log10 of number of C. albicans per gram of organ (with luminal contents) was 7.9 and 7.7 in the stomach and cecum, respectively. Direct fecal smears, as well as impresssion smears of stomach and cecum mucosal surfaces, revealed yeastphase cells, many with germ tubes, but no hyphal forms. No illness or mortality was observed over this period. The mice then were given amphotericin B DISsolved in the drinking water and offered ad libitum. At levels of 0.1 and 0.2 mg/ml, the number of fecal C. albicans was decreased but not eliminated completely. However, 0.3 mg/ml was sufficient to decontaminate the mice completely and return them to the germfree state. Residual amphotericin B was detected in the feces of the mice only while they were receiving the 0.3 mg/ml dose level. These mice remained germfree until the termination of the experiment, 10 weeks after the antibiotic had been discontinued and replaced by plain drinking water.

Administration, Oral↗

Effect of ethanol on phospholipid metabolism by the rat lung.

Prefeeding of alcohol shows the in vivo incorporation of orally administered palmitic acid into phosphatidylcholine of the lung. This impairment is also demonstrable in vitro utilizing lung slices and 14C-labeled palmitate or cytidine 5'-diphosphocholine as precursors. It is concluded that alcohol ingestion affects the utilization of precursors needed for phospholipid formation in the lung.

Animals↗

Chemical relaxation studies on the system liver alcohol dehydrogenase, NADH and imidazole.

Several years ago, Theorell and Czerlinski conducted experiments on the system of horse liver alcohol dehydrogenase, reduced nicotinamide adenine dinucleotide and imidazole, using the first version of the temperature jump apparatus with detection of changes in fluorescence. These early experiments were repeated with improved instrumentation and confirmed the early experiments in general terms. However, the improved detection system allowed to measure a slight concentration dependence of the relaxation time of around 3 ms. Furthermore, the chemical relaxation time was smaller than the one determined earlier (by factor 2). The data were evaluated much more rigorously than before, allowing an appropriate interpretation of the results. The observed relaxation time is largely due to rate constants in an interconversion of ternary complexes, which are faster than three (of the four) dissociation rate constants, determined previously by Theorell and McKinley-McKee.1,2 This fact contributed to earlier difficulties of finding any concentration dependence. However, the binding of imidazole to the binary enzyme-coenzyme complex can be made to couple kinetically into the interconversion rate of the two ternary complexes. The observed signal derives largely from the ternary complex(es). A substantial fluorescence signal change is associated with the observed relaxation process, suggesting a relocation of the imidazole in reference to the nicotinamide moiety of the bound coenzyme. Nine models are considered with two types of coupling of pre-equilibria (none-all). Quantitative evaluations favor the model with two ternary complexes connected by an interconversion outside the four-step (bimolecular) cycle. The ternary complex outside the cycle has much higher fluorescence yield than the one inside. The interconversion equilibrium is near unity for imidazole. If it would be shifted very much to the side of the "dead-end" complex (as in isobutyramide?!), stimulating action could not take place.

Alcohol Oxidoreductases↗

[Immunoelectronmicroscopic localization of cell wall antigens in streptococci. II. Localization of the group-specific polysaccharide of group C streptococci with ferritin- and peroxidase-labelled Helix pomatia-agglutinin (author's transl)].

The carbohydrate of the cell wall of group C streptococci is one of the best known group-specific streptococcal antigens with respect to its chemical structure. In the present paper, the ultrastructural location of the carbohydrate was investigated by means of immunoelectronmicroscopic techniques. Besides group-specific antibodies, the specific binding of an agglutinin (protectin Anti-AHP) of the edible snail (Helix pomatia) to structures with terminal N-acetylgalactosamine was used to demonstrate the antigen. The agglutinin was extracted from the albumen gland of the snail and purified by column chromatography. By means of glutardialdehyde it was coupled with ferritin or horesradish peroxidase. The investigations were done on strains of Streptococcus equisimilis and Streptococcus equi. Str. pyogenes (group A streptococci) was used as a control. On whole cells of group C streptococci the carbohydrate was demonstrated on the surface of the triple-layered cell wall. Near the cross-wall the carbohydrate seems to be more concentrated. In the presence of N-acety-D-galactosamine the labelling of the cell wall was inhibited. N-acetyl-D-glucosamine did not have such an effect. On isolated cell walls, both the outer and the inner surface were tagged. This suggests that the group-specific carbohydrate covers the peptidoglycan (mucopeptide) on both sides. The cytoplasmic membrane shows no reaction.

Agglutinins↗