Biomedical subjects
Publications and source records attributed to G Geyer.
Cytophotometric evaluation of mouse sperm glucose-6-phosphate dehydrogenase revealed a single peak-curve of enzyme activity. The finding is clearly contrary to the assumption of postmeiotic transcription of the X-chromosomal G6PDH-gene which was considered a possible source of increased enzyme activity in mature sperm cells.
With regard to recent findings of basic mechanisms of chromosomal functions the paper is concerned with the classification of the nucleus in the eukaryotic cell hierarchy. In conclusion, the nucleus is considered a cellular compartment a) temporary homing the main genetic substance, and b) providing facilities for appropriate regulation of its activity.
A 50-year-old female patient with a congenital cyst of the tongue is presented. Similar cases in literature are discussed as to their embryology, development and classification.
The present study is concerned with artifacts likely to occur in a horseradish peroxidase exclusion test. Incubation of murine peritoneal macrophages and lymphocytes with the peroxidase showed a close relationship between the number of living cells and the percentage of cells excluding the tracer. The penetration of the cytoplasm by horseradish peroxidase is attributed to an increase in the permeability of the cell membrane during the incubation (ranging from 10 to 120 min). It was not increased by the presence of tracer throughout the incubation period. However, concomitant fixation of the cell in the presence of horseradish peroxidase caused an increase in the influx of the tracer. The horseradish peroxidase exclusing test applied to the guinea-pig organ of Corti has proved to be valid provided that: (a) mechanical lesions prior to the tracer incubation are avoided; (b) incubation is terminated by removal of the extracellular tracer; (c) fixation is carried out as soon as possible; (d) a low concentration of horseradish peroxidase is used; and (e) specimens are incubated in diaminobenzidine-H2O2 medium for the shortest possible period. Although fixation-induced cytoplasmic infiltration by horseradish peroxidase was not detected in cochlear specimens, the findings call attention to possible sources of error and define the level of significance of the test. Horseradish peroxidase does not appear to be a cytotoxic agent under the conditions used.
The present study is concerned with the interaction of rat endo- and mesothelium with homologous erythrocytes under various conditions of pretreatment and incubation. Its findings show rat endo- and mesothelial cells a) nonadhesive to native or pretreated erythrocytes irrespective of the presence of gamma-globulin in the medium, b) devoid of primary or cryptic receptors sensitive to the Fc segment of the IgG molecule, and c) provided with binding sites at the oxidized glycocalyx which together with receptor groups of modified erythrocytes share the same class of IgG.
The study reports on plasmalemmal perturbation of erythrocytes under osmotic stress. In comparison with isotonic human erythrocytes a 10 minutes incubation in 150 mosm sodium cacodylate pH 7.4 at 20 degrees C results in an increase of the anti AH P binding rate by a factor of two. 0.1 M N-acetyl galactosamine inhibits the binding of anti-AH P of both isotonic and hypotonic red cells. Contrary to isotonic mouse erythrocytes the agglutination with peritoneal murine leukocytes of hypotonic red cells is greatly enhanced provided the erythrocytes have been incubated in autologous serum. The findings are considered evidence of the unmasking of binding sites due to nonspecific rearrangement of membrane constituents.
Plasmalemmal differentiation of the enucleating normoblast of rabbit and rat was studied by means of cytochemical methods and freeze-etching. Staining with colloidal iron revealed about identical amounts of iron particles bound to various areas of the normoblast membrane. Cationized ferritin and ruthenium red, likewise, failed in the demonstration of significant changes of the enucleating normoblast glycocalyx. Despite these findings the topo-optical staining with toluidine blue showed the plasmalemmal envelope of the protruding normoblast nucleus moderately birefringent, clearly discriminated from the intense anisotropic staining of the future reticulocyte membrane. The ferritin-labeled snail lectin anti AHP localized a great number of binding sites at the plasmalemmal envelope of the nucleus under extrusion. That is in sharp contrast with rather low lectin binding to the future reticulocyte membrane which amounts to about 30 to 50% of the nuclear envelope label. The findings provide evidence of unmasking of bindings sites of the normoblast membrane. Apparently, the effect is due to conformational changes of the cell membrane, rather than it could be attributed to degradation of glycoproteins. Moreover, enucleation kinetics may also be related to supramolecular changes of membrane structure albeit missing evidence for the rearrangement of membrane particles.
A cetylpyridinium chloride-phosphotungstic acid procedure suggested by Kelényi and Kiss (1976) for the staining of mast cell granules in ultrathin sections was slightly modified with regard to pretreatment, pH of incubation, and rinsing of sections. The method also proved suitable for the demonstration of various mucosubstances and of cartilage proteoglycan aggrebates.
Electron microscopic findings of this study revealed 2 types of extrusion of brush border cells from the summits of intestinal villi: a) By expansion of initial vacuolation the degenerating cell enlarges and elevates itself above the level of the extrusion zone. Cells about to lift become sphered and are shed into the lumen for degradation to continue. b) Another group of cells vacuolating to a similar extend already may start apical decay in situ. In this case mechanical elevation, apparently, is inefficient or retarded such as to offer a cell the opportunity to decompose at its proper place in the mucosal lining. Contrary to type A extrusion, i.e. the desquamation of the entire degenerating cell, type B produces cellular debris to be shed into the lumen. Either type of exfoliation may be associated with the formation of gaps the closure of which proceeds slowly. The light microscopic test of cell viability by means of horseradish peroxidase demonstrated many brush border cells leaky such as to become loaded with the tracer irrespective of its vascular or enteric approach to the extrusion zone. Damage of the cell membrane due to a brief pretreatment with digitonin resulted in a dramatic rise of horseradish peroxidase positive cells. With respect to these findings, one cannot escape the conclusion that both types of extrusion confer intermittent leakage on the summits of intestinal villi which at least could account for part of the diffusion of macromolecules in either direction.
The present study applies semithin sections for electron microscopic autoradiography. Due to (a) the decrease of exposition time, and (b) lower intensity of isotope labelling, low power electron microscopic autoradiography on semithin sections is favoured in comparison with conventional (ultrathin section high power) electron microscopic autoradiography, and due to (c) more precise localization of silver granules, and (d) ease of quantitative evaluation of autoradiography, it is superior to ordinary (thick section) light microscopic autoradiography.
The effect on the net negative surface charge of single or multiple coating of erythrocytes was studied on human group A red cells previously washed and fixed. Single coating with either group A antiserum or protectin decreased the electrophoretic mobility of erythrocytes by 16% and 13%, respectively. Following an incubation in group A glycophorin or bovine submaxillary mucin of agglutinin coated cells, the electrophoretic mobility was restored or slightly increased compared with uncoated erythrocytes. Upon repeating this scheme, the second agglutinin binding again reduced these cells' cytophoretic mobility, and they were accelerated once more by a second coating with acid glycoprotein. Intensity and homogeneity of colloidal iron staining of uncoated and variously coated erythrocytes reflected their electrophoretic retardation and acceleration, respectively.
Fractions of light and heavy erythrocytes were separated by centrifugation from blood samples banked in ACD-AG medium at 4 degrees C for periods up to 6 weeks. Both light and heavy erythrocytes have lost about 4,9% of their content of sialic acids during banking for 6 weeks. This reduction is in accord with a 6%-decrease of their agglutination by means of alcian blue. It is, however, a variance with the inhibition of agglutination by anti-IgG sera. The present findings provide evidence for the role alterations of the red cell membrane may play in the process of recognition and phagocytosis of banked erythrocytes. With regard to these alterations we suggest two types of rapid elimination of transfused banked erythrocytes: a) Primary elimination refers to cells primarily loaded with immunoglobulins such as to get recognized and phagocytized by macrophages. b) Secondary elimination accounts for rigid erythrocytes suffering from additional degradation while retained in the spleen prior to their loading with immunoglobulins and ensuing phagocytosis. Secondary elimination is considered a process more relevant to reutilisation of banked blood.
We suggest a method for the section staining of polyanions by means of ruthenium red and osmium tetroxide. A 5 min incubation in diluted ruthenium red-OsO4 solution pH = 7.4 of ultrathin Durcupan-sections previously treated with sodium methoxide results in the opaque stining of mast cell granules, cartilage proteoglycans, and various epithelial mucosubstances.
Rapid loss of cochlear microphonics in guinea-pigs previously exposed to high-energy impulse noise was shown to be related to the breakdown of the endolymphatic boundary. The cochlear duct was rendered leaky by deterioration of the reticular membrane, and damage of sensory and supporting cells.
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Selective binding of toluidine blue to basophilic proteoglycans is the first stage of a staining method which proceeds to the formation of a heavy metal salt of the dyestuff. By means of this procedure matrix complexes of hyalin cartilage were stained such as to display granular subunits about 25 to 35 A in diameter. The findings are interpreted as the demonstration of proteoglycan constituents of chondromucin aggregates.
Fixation with Ca2+ -glutaraldehyde of ghosts results in opaque membrane associated deposits similar to Ca2+ binding sites of native human erythrocytes. Following brief incubation in an ATP medium the number and size of major Ca2+ affinity sites is considerably enhanced. In addition to major Ca2+ affinity sites multiple minor sites are lining either aspect of the ghost membrane. Ghosts fixed with EDTA-glutaraldehyde are devoid of major Ca2+ affinity sites and they exhibit extreme low overall opacity. Ghosts previously partially despectrinated by incubation in 0.5 mM EDTA have lost major Ca2+ affinity sites, although minor binding sites appear unimpaired. The findings provide evidence of the demonstration of phosphorylated spectrins in major Ca2+ affinity sites.