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Biomedical subjects

W Buchheim

Publications and source records attributed to W Buchheim.

10 recordsLinked to original sources

Histochemical and biochemical observations on milk-fat-globule membranes from several mammalian species.

A specific secretory product of the lactating mammary gland are triglyceride fat globules which are enveloped by a very complex membrane, the milk fat globule membrane (MFGM). In different mammalian species (man, rhesus monkey, horse, goat, sheep, cow, grey seal, camel, alpaka) the glycoproteins of this membrane have been analyzed by gel electrophoresis, Western blotting and lectin histochemistry. A remarkable intra- and interspecific variability of these glycoproteins has been detected pointing to so far unknown physiological adaptions, which may play a role in the intestine of the new born. High molecular weight glycoproteins, with a very high degree of glycosilation have been found only in primates, horse and camel; the MFGMs of the true ruminants (cow, sheep, goat) are characterized by specific glycoproteins of a lower molecular weight range.

Animals

Structural, histochemical and biochemical observations on horse milk-fat-globule membranes and casein micelles.

Horse milk fat globules (MFGs) and casein micelles were studied using freeze fracturing, freeze etching and thin-section electron microscopy, as well as lectin histochemistry, gel electrophoresis, and Western blotting. Horse MFGs were found to be relatively small, their average volume-surface diameter being about 2.75 microns. The MFG membrane is composed of three layers: an inner proteinaceous coat occasionally having a paracrystalline substructure, a unit membrane, and a prominent filamentous glycocalyx. The last is rich in glycoconjugates, as revealed by its binding of various lectins. In addition, the glycocalyx binds cationized ferritin, which indicates the presence of negative electric charges. Gel electrophoresis revealed the presence of high-molecular-weight glycoproteins in the MFG membrane of horse milk. Such glycoproteins are also present in human MFG membranes but are absent in the bovine MFGs. The casein micelles in horse milk are relatively large, their average volume-surface diameter being about 200 nm.

Animals

Glycoprotein filament removal from human milk fat globules by heat treatment.

Freeze-etch electron microscopy was applied to milk fat globules to observe surface details. A remarkable array of filaments, approximately 0.5 micron in length, was seen on human, but not bovine, globules. Heating human globules removed the filaments that were identified as high molecular weight glycoproteins by freeze-etch and gel electrophoretic analysis of the heating medium. Extraction of these globule glycoproteins was slight at 60 degrees C for one minute but substantial and tending to plateau at 80 degrees C for one minute. Such heat-induced alterations of the milk fat globule surface provide an alternative or additional explanation to milk lipase inactivation as the cause of reduced milk fat absorption from heated milk by the preterm infant.

Animals

Freeze-fracture studies of cytoplasmic inclusions occurring in experimental lipidosis as induced by amphiphilic cationic drugs.

The ultrastructure of cytoplasmic inclusions, which characterize experimental lipidosis as induced by several amphiphilic cationic drugs, was studied by means of freeze-fracturing and thin-sectioning. Retinal and adrenal tissues of rats chronically treated with high oral doses of chlorphentermine were used. In thin sections the cytoplasmic inclusions, which were previously shown to represent lysosomes overloaded with polar lipids, exhibit lamellated or lattice-like internal patterns. The present freeze-fracture observations are interpreted as to indicate that the lamellated inclusions contain polar lipids in the lamellar phase, whereas those with lattice-like patterns contain polar lipids in a hexagonal phase.

Adrenal Glands

On the size of monomers and polymers of beta-casein.

The size and number average molecular weight have been determined for beta-casein monomers and polymers from electron micrographs using the freeze-etching procedure with spray-frozen specimens. For the spherical beta-casein monomers we found a mol. wt of 22600 and a diam. about 10 nm, which compared quite well with data obtained from ultracentrifugation, light scattering and viscosity measurements. Polymer sizes were in agreement with molecular weight determinations from ultracentrifugation and light scattering, assuming that the volume and weight of the particles are proportional.

Caseins

The functional and structural border between the CSF- and blood-milieu in the circumventricular organs (organum vasculosum laminae terminalis, subfornical organ, area postrema) of the rat.

The present study continues a previous investigation on the median eminence (EM) (Krisch et al., 1978). In rats with high levels of neurohormones (LHRH, vasopressin) a limited immunohistochemical labeling of perivascular tanycyte processes can be observed surrounding capillaries in the marginal region of the organum vasculosum laminae terminalis (OVLT) and in the inner part of the subfornical organ (SFO). This labeling extends from the perivascular space a short distance along the tanycyte processes. By conventional electron microscopy and by freeze-etching, tight junctions are demonstrated at a distance from the capillary lumen which corresponds to the borderline of the immunohistochemical labeling of perivascular tanycyte processes in light microscopic preparations. The tight junctions are arranged in several parallel and helical rows and correspond to those found in the median eminence. Consequently, the immunohistochemical labeling the OVLT and in the SFO marks the intercellular cleft. In the circumventricular organs the immunostaining labels the extension of the perivascular space characterized by the hemal milieu. The perivascular space is separated off by tight junctions from the CSF-milieu of the adjacent neuropil. Furthermore, the present study demonstrates tight junctions in the marginal region of the area postrema (AP) between the perivascular processes of the tanycytes.

Animals

Freeze fracture studies on the annelid septate junction.

Freeze-fractured preparations of septate junctions between epidermal cells of annelids (Lumbricus terrestris and Tubifex spec.) have been investigated. In Lumbricus the protoplasmic face (PF) of the plasma membrane is characterized by variously arranged rows of particles. Apically the rows take an undulating course and often are separated by wide distances. In the basal part of the junction the rows run closely together and more or less in parallel. The diameter of the particles measures 80--120 A, the distance between two particles (centre to centre) is 150--250 A. Additionally striking rows of large particles (long diameter 150--200 A). Are to be observed mainly near the basal part of the junction. In Tubifex both faces of the plasma membrane could be studied in detail. The protoplasmic face (PF) contains rows of distinct individual particles (mean diameter 100--150 A, centre to centre distance approx. 250 A) whereas the particles of the extracellular face (EF, mean diameter 200-250 A) usually form continuous strands in which the individual particles seem to fuse. The density of arrangement of the strands varies considerably. Additionally ladder-shaped membrane structures have been observed in plasma membranes of this species.

Animals

Freeze-etching of unglycerinated tissue dispersions by application of the oil emulsion technique.

The oil emulsion technique has been applied to rat brain and liver tissue for freeze-etching preparation without antifreeze pretreatment. It has been demonstrated that the dispersed tissue exhibits a high degree of vitrification throughout the mostly globular fragments up to at least 30 micrometer in diameter. Signs of insignificant ice crystal formation of 50 nm and less could only rarely be detected. Although the dispersion process seriously affects the tissue organization as a whole, most cellular organelles and the plasma membrane show satisfactory preservation of structure. In larger tissue drops, a local overall structural preservation of the original cellular organization is recognizable. The results obtained demonstrate the applicability of this method to various biological objects for realizing sufficient cryofixation in the absence of cryoprotectants.

Animals