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Biomedical subjects

W T Daems

Publications and source records attributed to W T Daems.

At least 55 records · Page 3Linked to original sources
PubMed1981

The effect of chloroquine on lysosomal function and cell-coat glycoprotein transport in the absorptive cells of cultured human small-intestinal tissue.

The effect of chloroquine, an inhibitor of intralysosomal catabolism, on the synthesis, transport, and degradation of cell-coat glycoproteins in absorptive cells of cultured human small-intestine tissue was investigated by morphometrical, autoradiographical, and biochemical methods. Neither synthesis nor transport of cell-coat material was affected by the drug, but culturing of the absorptive cells in the presence of chloroquine led to a dose- and time-dependent enlargement of the dense bodies; other cell structures showed no alterations. 3H-fucose-labelled material accumulated in the dense bodies of the absorptive cells of these cultures. Since no increase of beta-glucuronidase and acid phosphatase activity (both lysosomal enzymes of glycoprotein nature) was found, this accumulation of radiolabeled material can be explained as a chloroquine-mediated inhibition of the degradation of cell-coat glycoproteins. These macromolecules probably enter the lysosome-like bodies by a crino-phagic mechanism, i.e., fusion of these organelles with the apical vesicles and tubules involved in intracellular transport. These findings suggest that the lysosome-like bodies have a function in the regulating of cell-coat glycoprotein transport in human intestinal absorptive cell, i.e., the degradation of excess cell-coat material.

Biological Transport↗
PubMed1981

Endocytosis in absorptive cells of cultured human small-intestinal tissue: horseradish peroxidase, lactoperoxidase, and ferritin as markers.

The occurrence of endocytotic mechanisms in human small-intestinal absorptive cells was investigated by culturing biopsy specimens in the presence of horseradish peroxidase (HRP), lactoperoxidase (LPO), and ferritin. The results indicate that both HRP and LPO entered the cells by apical endocytosis, after which they were transported via apical vesicles and tubules to the lysosome-like bodies. Ferritin, which showed a distinct affinity for the cell-coat glycoproteins, was not interiorized by the absorptive cells. These findings suggest that although human absorptive cells have an endocytotic mechanism, possibly fluid-phase endocytosis, cell-coat glycoproteins are not taken up by the cells, as indicated by the absence of ferritin in the apical vesicles and tubules, as well as the lysosome-like bodies. These findings provide indirect support for our hypothesis that the lysosome-like bodies have a function in the regulation of cell-coat glycoprotein transport via a crinophagic mechanism (fusion of apical vesicles and tubules with lysosome-like bodies) rather than via an exocytotic-endocytotic mechanism.

Cells, Cultured↗
PubMed1981

The effect of colchicine on the intracellular transport of 3H-fucose-labelled glycoproteins in the absorptive cells of cultured human small-intestinal tissue. An autoradiographical and biochemical study.

The effect of colchicine on the intracellular transport of 3H-fucose-labelled glycoproteins in the absorptive cells of cultured biopsy specimens of the human intestine was investigated by light- and electron-microscopical autoradiography and by biochemical methods. The results showed a decrease in the radioactivity of the cell coat on the microvilli and an increase in the Golgi apparatus and in the apical vesicles and tubules. This divergence is attributed to a colchicine-induced impairment of the normal transport of cell-coat glycoproteins from the Golgi apparatus, via the apical vesicles and tubules, to the apex of the cell. The radioactivity of the lysosome-like bodies in the absorptive cells cultured with colchicine also increased. This finding supports a crinophagic function of these organelles in the degradation of excess cell-coat material.

Biological Transport↗
PubMed1981

The effect of chloroquine on the intralysosomal degradation of cell-coat glycoproteins in the absorptive cells of cultured human small-intestinal tissue as shown by silver proteinate staining.

The effect of chloroquine on the intralysosomal degradation of cell-coat glycoproteins in cultured intestinal absorptive cell was investigated by silver proteinate staining. The results of this staining method, which is specific for carbohydrate containing macromolecules such as glycoproteins and mucopolysaccharides, showed that in the presence of the drug considerable amounts of silver proteinate-positive material accumulated in one type of lysosome-like body: the dense bodies. The staining pattern of other cell organelles was not affected by chloroquine. The presence of the drug in the culture medium also resulted in the occurrence of numerous small vesicular structures in the matrix of the dense bodies. These showed a similar size and structure to those present in the other type of lysosome-like body: the multivesicular bodies. This observation, together with earlier autoradiographical data, suggests that cell-coat material is transferred from multivesicular to dense bodies by fusion between these organelles. This study thus provides further evidence for a regulatory mechanism of cell-coat glycoprotein transport by the lysosome-like bodies in human intestinal absorptive cells.

Chloroquine↗
PubMed1981

Heterogeneity in wheat germ agglutinin binding by mouse peritoneal macrophages.

The binding of the lectin wheat germ agglutinin (WGA) to the cell surface of monocytes and macrophages obtained from the stimulated peritoneal cavity of mice was investigated electron microscopically, using ovomucoid-gold as an indirect marker. Resident (tissue) macrophages, identified by the presence of PO activity in the rough endoplasmic reticulum as well as in the nuclear envelope, showed low WGA binding, whereas monocytes and monocyte-derived macrophages with PO activity in the granules showed high WGA binding. Since cells devoid of PO activity showed variable WGA binding, the value of this gold-WGA-binding technique for discrimination on a quantitative basis between resident macrophages and monocytes or monocyte-derived macrophages, is discussed.

Animals↗
PubMed1980

Binding of cationized ferritin to the cell-coat glycoproteins of human and rat small-intestinal absorptive cells.

The binding of cationized ferritin (CF) to the cell-coat (glycocalyx) glycoproteins of human and rat intestinal absorptive cells was investigated in relation to the amount of sialic acid in these macromolecules. The cell coat of human absorptive cells exhibited poor binding of CF and contained a small amount of sialic acid. The cell coat of rat absorptive cells had about ten times more sialic acid than that of human cells and showed a strong affinity for the marker. The removal of sialic acid from the cell-coat glycoproteins of rat intestinal cells by neuraminidase treatment abolished CF binding. These results suggest that sialic acid is necessary for CF binding and that human and rat intestinal absorptive cells show a species-specific difference in the sugar composition of the cell coat.

Animals↗
PubMed1979

Resolution of a gold latensification-elon ascorbic acid developer for Ilford L4 emulsion.

The electron-microscopical autoradiographical resolution of a gold latensification-elon ascorbic acid (GEA) developer for Ilford L4 emulsion was determined experimentally, using radioactive line sources of tritiated albumin (Heijnen and Geuze, 1977). For sections with a thickness of 62 nm (SD +/- 11), which were covered with a carbon layer about 5 nm thick and a slightly overlapping monolayer of L4 silver bromide crystals, the measured half-distance (HD) of resolution was 115 nm. This improvement in resolution, the high efficiency of the GEA developer for L4 emulsion (Wisse and Tates, 1968), and the excellent visibility of the cellular structures under the small silver grains, mean that the L4-GEA combination deserves preverence as a method for quantitative electron-microscopical autoradiography.

Ascorbic Acid↗
PubMed1979

Transport of radiolabelled glycoprotein to cell surface and lysosome-like bodies of absorptive cells in clutured small-intestinal tissue from normal subjects and patients with a lysosomal storage disease.

The transport of 3H-fucose- and 3H-glucosamine-labelled glycoproteins in the absorptive cells of cultured human small-intestinal tissue was investigated with light- and electron-microscopical autoradiography. The findings showed that these glycoproteins were completed in the Golgi apparatus and transported in small vesicular structures to the apical cytoplasm of these cells. Since this material arrived in the cell coat on the microvilli and in the lysosome-like bodies simultaneously, a crinophagic function of these organelles in the regulation of the transport or secretion of cell-coat material was supported. In the absorptive cells of patients with fucosidosis or Hunter's type of lysosomal storage disease, a smiliar transport of cell-coat material to the lysosome-like bodies and a congenital defect of a lysosomal hydrolase normally involved in the degradation of cell-coat material, can explain the accumulation of this material in the dense bodies.

Adolescent↗
PubMed1979

The differentiation of monocytes into macrophages, epithelioid cells, and multinucleated giant cells in subcutaneous granulomas. I. Fine structure.

The morphological changes occurring in monocytes during their differentiation into macrophages, epithelioid cells, Langhans-type giant cells, and foreign-body-type giant cells were investigated in foreign-body granulomas induced by subcutaneous implantation of pieces of Melinex plastic. Analysis based on Adams's (1974) criteria for discrimination between the several types of cell of the monocyte line, showed that each type has a characteristic type of granule. Primary and secondary granules, numerous in the Golgi area of monocytes were generally found close to the cell membrane and decreased in number in maturing macrophages. This was accompanied by an increase in the number of microtubules. Mature macrophages show numerous characteristic macrophage granules, which are round (average diameter: 280 nm) and have a halo between the limiting membrane and granular matrix. Mature epithelioid cells have characteristic epithelioid cell granules, and multinucleated giant cells a heterogenous population of granules. Fusing macrophages generally have their Golgi areas facing each other, and also show a reduced thickness of the cell coat. The morphology of the multinucleated giant cell is closely related to the number of nuclei present. In Langhans-type giant cells, which generally have two to ten nuclei, a giant centrosphere with numerous aggregated centrioles is found. In transition forms between Langhans-type and foreign-body-type giant cells, which generally contain 10--30 nuclei, the centrioles show less aggregation. In the foreign-body-type giant cells, which generally have more than 30 nuclei, centrioles are virtually absent and never aggregated. These differences between the Langhans-type giant cells, the foreign-body-type giant cells, and the transition forms, support our previous finding that Langhans-type giant cells are the precursors of foreign-body-type giant cells.

Animals↗
PubMed1979

The differentiation of monocytes into macrophages, epithelioid cells, and multinucleated giant cells in subcutaneous granulomas. II. Peroxidatic activity.

The peroxidatic (PO) activity of monocytes differentiating into macrophages, epithelioid cells, and multinucleated giant cells in subcutaneous granulomas was investigated with three different media for the demonstration of PO activity. Irrespective of the stage of differentiation, these cells did not show PO activity in the rough endoplasmic reticulum (RER) or nuclear envelope. In addition, it was found that the morphologically characteristic types of granule of the various cells of the monocyte line (the primary granules and secondary granules of monocytes, the macrophage granules, and the epithelioid cell granules), all have distinct cytochemical characteristics. Monocytes lose their primary and secondary granules during differentiation into mature macrophages. Simultaneously, the granules of both types become elongated and the secondary granule lose their halo. In contrast to monocytes, mature macrophages may contain a few microperoxisomes. During the differentiation of macrophages into epithelioid cells or multinucleated giant cells there is an increase in the number of microperoxisomes.

Animals↗
PubMed1979

Comparative study on peroxidatic activity in inflammatory cells on cutaneous and peritoneal implants.

Inflammatory reactions were evoked by simultaneous implantation of pieces of Melinex plastic in the subcutaneous tissue of the dorsum and in the peritoneal cavity of rats. The cellular composition of the Melinex-adherent cells and their peroxidatic (PO) activity were investigated in relation to the duration of implantation. Several striking differences were found between the subcutaneous and peritoneal implants. On the 7th and 14th days, multinucleated giant cells were abundantly present on the subcutaneous implants, whereas they were relatively rare on the peritoneal implants. The subcutaneous implants bore no mast cells and only a few eosinophilic granulocytes, but both types of cell were observed frequently on the peritoneal implants. Macrophages and multinucleated giant cells on the subcutaneous implants show PO activity only in the granules are PO negative. On the peritoneal implants three types of macrophages can be dintinguished: exudate macrophages which have PO activity restricted to granules or are PO-negative; macrophages with PO activity in granules and both the rough endoplasmic reticulum (RER) and nuclear envelope; and resident macrophages with PO activity only in the RER and nuclear envelope. In addition, two types of multinucleated giant cells are found, one with and the other without PO activity in the RER and nuclear envelope. Multinucleated giant cells with PO activity in the RER and nuclear envelope as well as exudate macrophages with PO activity in the RER and nuclear envelope were mainly found 32 h and 3 days after implantation of the Melinex in the peritoneal cavity. These findings are discussed in the light of current knowledge of the PO activity in macrophages and multinucleated giant cells. It is concluded that the appearance of PO activity in the RER and nuclear envelope of exudate macrophages and multinucleated giant cells is in all probability a transient phenomenon, and that there is no objective evidence to support the opinion that exudate macrophages with PO activity in the RER and nuclear envelope are transitional cells between exudate and resident macrophages.

Animals↗
PubMed1979

Fusion of erythrolysosomes in epithelial cells of the sheep placenta.

In trophoblastic epithelial cells of the sheep placenta the breakdown of erythrocytes within complex erythrolysosomes was studied at the ultrastructural level. It was found that the formation of complex erythrolysosomes containing from two to several erythrocytes as a result of fusion of erythrolysosomes within the epithelial cells was a common occurrence when the epithelial cells engulfed a large number of erythrocytes. The erythrocytes enclosed in complex erythrolysosomes appear to be either in the same or in different stages of hemolysis. In the process of breakdown of erythrocytes within complex erythrolysosomes five successive stages of hemolysis could be distinguished. Acid phosphatase activity was demonstrated in the complex erythrolysosomes and appeared to be located in the angular interspaces between the erythrocytes and the lysosomal membrane. The fragmentation of complex erythrolysosomes with formation of small hemoglobin-containing lysosomes also occurred. The fusion of erythrolysosomes with formation of complex erythrolysosomes can be considered as an additional mechanism in the process of erythrocyte breakdown in the epithelial cells of the sheep placenta.

Acid Phosphatase↗
PubMed1979

A new dynamic model system for the study of capture reactions for diffusable compounds in cytochemistry. II. Effect of the composition of the incubation medium on the trapping of phosphate ions in acid phosphatase cytochemistry.

A model system developed for the study of the dynamics of capture reactions for diffusable compounds in cytochemistry served as a basis for the experiments reported in the present paper. The model was used to study the effect of the composition of the cytochemical medium on the trapping of phosphate ions by lead (II) ions in acid phosphatase cytochemistry. In this system a phosphate-containing solution and a lead-containing solution (cytochemical medium) are pumped along opposite sides of a polyacrylamide film. The phosphate concentration at which measurable precipitation starts in the film (critical phosphate concentration) was taken as a measure of the trapping efficiency of the cytochemical medium. The addition of beta-glycerophosphate and cytidine-5'-monophosphate to a buffered lead-containing solution resulted in a higher critical phosphate. The addition of chloride ions and acetone, as well as decreasing the molarity of the acetate buffer of the cytochemical medium, were found to lower the critical phosphate concentration, whereas the addition of fluoride ions, glucose, and sucrose had no effect. From the effect of variations in the composition of the cytochemical medium on the trapping efficiency and the turnover number of acid phosphatase in the medium, it was possible to predict which cytochemical medium would be the most suitable for the demonstration of acid phosphatase activity in guinea-pig peritoneal exudate cells. The results were in accordance with the localization of acid phosphatase activity: the higher the trapping efficiency and the turnover number, the higher the amount of precipitate and the number of positive enzymatic sites. In this way an improved cytochemical medium for acid phosphatase was developed.

Acetone↗
PubMed1978

The ultrastructure of guinea pig heterophil granulocytes and the heterogeneity of the granules.

The development of the heterophil granulocytes in the bone marrow of the guinea pig is described. During the maturation of these cells, three types of granule are formed, not only the azurophil and specific granules already described in other mammals but also a third type of granule referred to here as the nucleated granule. During the process of maturation of the cells, these three types of granule are formed successively. On this basis, two steps can be distinguished in the promyelocyte phase in which primary (nucleated and azurophil) granules are formed, i.e. an early and late stage, nucleated granules being formed in early and azurophil granules in late promyelocytes. Secondary (specific) granules occur first in myelocytes. In mature heterophils of the guinea pig the granule population is composed of about 85% secondary granules, about 10% azurophil granules, and about 5% nucleated granules. The changes in the granule population during the maturation process were quantified. The observations and calculations point to the occurrence of three mitoses: one in the early and one in the late promyelocyte and the third in the myelocyte.

Animals↗