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Biomedical subjects

B Voss

Publications and source records attributed to B Voss.

At least 91 records · Page 5Linked to original sources

Temperature dependence of lipogenesis in isolated hepatocytes from rainbow trout (Salmo gairdneri).

Temperature dependence of lipogenesis in trout liver cells was investigated in the presence of 5 mM lactate using either [14C]lactate or [3H]water. A ratio of 3H/14C-incorporation greater than one is found, irrespective of temperature. Acclimation of fish to 4, 10 or 16 degrees C affects neither the height of lipid synthesis nor its temperature sensitivity. The distribution of [14C]lactate between the main lipid classes and the capacities for cholesterol- and triacylglycerol-synthesis are correlated to the glycogen stores of the hepatocytes. A comparison of fatty acid synthesis and cholesterogenesis in livers of normal fed rat and of trout suggests a capability for lipogenesis in trout somewhat similar to that in mammals.

Acclimatization↗

Localization of collagen types I, III, IV and V, fibronectin and laminin in human arteries by the indirect immunofluorescence method.

The distribution of types I, III, IV and V collagen and of the glycoproteins fibronectin and laminin in sections of human aortas, arteries and atherosclerotic plaques were studied using monospecific antibodies and indirect fluorescence microscopy. Types IV and V collagen and laminin were present in a narrow zone, representing the basement membrane, apposed to the endothelial layers of all these tissues. Types I and III collagen and fibronectin were located in the interstitial spaces of the intima and the media of blood vessels walls, whereas types IV and V collagen and laminin were found in the basement membranes underlying smooth muscle cells in these areas. Two types of atherosclerotic plaques were observed. Lipid-rich plaques contained less collagen and reduced amounts of the glycoproteins. Fibrous plaques consisted of regions deficient in types I and III collagen and collagen-rich regions with elevated levels of these two collagens as well as more fibronectin. The collagen-rich regions of fibrous plaques contained, however, little type IV and type V collagen and little of the glycoproteins laminin and fibronectin. This may be due to the reduced number of cells involved in the biosynthesis of these basement membrane proteins.

Aorta↗

Basement membrane components (7S collagen, laminin P1) are increased in sera of diabetics and activate platelets in vitro.

Serum concentrations of two basement membrane components (7S collagen and laminin P1) were detected by specific radioimmunoassays in 70 patients suffering from diabetes mellitus type I and II with and without clinical signs of microangiopathy. Serum levels of both antigens were increased compared to controls. 7S collagen concentrations were significantly different between the diabetics with signs of microvascular damage and those without small-vessel disease (p less than 0.05). Laminin P1 concentrations were also elevated, but the difference between the two groups of diabetics was not significant (p less than 0.2). Raised levels of circulating basement membrane proteins may indicate connective tissue activity and development of diabetic microangiopathy. In vitro 7S collagen is a moderate platelet activator inducing platelet spreading, aggregation, and malondialdehyde production. Laminin activates platelet spreading. As a part of the altered hemostatic system the activation of basement membrane components may contribute to the development of microvascular damage.

Adult↗

Immunocytochemical investigation on the distribution of small chondroitin sulfate-dermatan sulfate proteoglycan in the human.

Polyclonal antibodies against the core protein of the small chondroitin sulfate-dermatan sulfate proteoglycan from human skin fibroblast secretions were used, after affinity-purification, as a probe to study localization of crossreactive material in several human tissues by indirect immunocytochemistry. In contrast to skin, kidney, and the adventitial layer of aorta, positive staining of brain, liver, cartilage, and intimal and medial layers of aorta required pre-treatment of tissue sections with chondroitin ABC lyase. In all tissues investigated, antigenic material was present in the interstitial space. Filamentous structures were perpendicularly oriented towards basement membranes. In liver, specific staining was seen along the sinusoidal walls. Reticular fibers with or without focal condensations were seen in cerebral cortex and cerebellum. The results suggest a role of small chondroitin sulfate-dermatan sulfate proteoglycan in cell-matrix interactions.

Animals↗

Synthesis, intracellular processing and secretion of thrombospondin in human endothelial cells.

The biosynthesis of thrombospondin, a glycoprotein first described in platelets, has been studied in human endothelial cells. This glycoprotein has a molecular mass of 450 kDa. It is secreted and incorporated into the extracellular matrix of several cell types in culture. Pulse-chase experiments with [3H]leucine were performed and the synthesis and secretion of the glycoprotein was studied by immunoprecipitation and sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The results of these experiments show that the three subunits of thrombospondin are identical in molecular mass. During synthesis there is a small but significant increase in molecular mass within 20 min after pulse labeling. The early form of thrombospondin is sensitive to endoglucosaminidase H treatment, indicating that a transformation of the oligosaccharide structures from 'high-mannose' to 'complex' structures takes place. Within 60 min after synthesis only the mature form of the glycoprotein is secreted into the medium. In the presence of tunicamycin, an inhibitor of N-glycosylation, there is a reduction in molecular mass of the subunit from 165 kDa to 155 kDa. Pulse-chase experiments in the presence of tunicamycin supported the conclusion that the carbohydrate part is processed during biosynthesis. Inhibition of glycosylation had a pronounced effect on the secretion of thrombospondin. The decreased occurrence of thrombospondin in the culture medium seemed to be due to a high intracellular degradation rate of unglycosylated thrombospondin. Characterization of the glycopeptide structures of thrombospondin metabolically labeled with [3H]mannose by Bio-Gel P-4 and concanavalin-A-Sepharose column chromatography revealed that the oligosaccharide structures of the cellular and secreted forms of thrombospondin differ in their composition.

Acetylglucosaminidase↗

A double antibody sandwich microELISA for measuring human platelet factor 4.

With the aim of investigating patients with a higher risk of thrombosis we developed a method for determining platelet factor 4 (PF4) in human plasma. Using the double antibody sandwich ELISA technique we set up a test system that allows the determination of PF4 concentrations in plasma samples from 1 to 100 ng/ml. This method is more sensitive and as specific as commercially available RIA kits, but has the advantage of being cheaper and less time consuming. Furthermore the ELISA does not require radioactive materials. The complete reaction is carried out in microtiter wells, and an ELISA-reader connected to an Apple computer does all the calculations needed for quantitative measurements.

Blood Specimen Collection↗

Connective tissue proteins on the injured endothelium of the rat aorta.

Type V collagen (TVC), fibronectin (FN), and laminin (LAM) were detected on the endothelial surface of mechanically injured rat aortas with the help of monospecific antisera and protein A - gold conjugates, carbon film surface replicas, and conventional embedding techniques. Deendothelialized tracks were produced in the thoracic aorta, and the presence of the connective tissue proteins on the luminal surface of the endothelium was studied. The changes in the distribution of the proteins during repair of the endothelial surface was followed for up to 6 days after injury. From 1 to 3 days after injury small numbers of gold particles, indicating the presence of TVC, were found between the adherent platelets on the freshly deendothelialized subendothelial matrix and in higher amounts on cell debris and collagen fibers. On the sixth day after injury, however, the amount of TVC between the sparsely distributed platelets on the deendothelialized areas was significantly higher than it was previously. FN and LAM were readily detectable on the subendothelial matrix and on the damaged marginal endothelial cells. These proteins were especially obvious on both margins of the tracks even from the first day after treatment. FN was found also in connection with fibrin precipitations as well as on the surface of some platelets and monocytes. The amount of FN and LAM present on the damaged area decreased slightly up to the sixth day. Monocytes and leukocytes adhered mostly at the margin of the wound area in the vicinity of the lesions on the endothelium. FN and LAM were often detectable under and around these adherent cells. Little of the connective tissue proteins was found on the uninjured and on the regenerated endothelial cells. The results showed subtle transitory changes in the surface pattern of the subendothelial connective tissue matrix of the injured intima. The adhesion of blood-borne cells may have been induced by FN and LAM on the endothelial surface near the lesions, and later partly prevented by increasing amounts of TVC on the surface.

Animals↗

7 S collagen: a method for the measurement of serum concentrations in man.

A radioimmunoassay was developed for the detection of 7 S collagen, a basement membrane component derived from type IV collagen, in sera of patients. Two forms of 7 S collagen were isolated by limited collagenase digestion of type IV collagen from bovine placenta. Antisera against both forms were raised in rabbits. The antigens were labeled by conjugation with the Bolton Hunter reagent or by the Chloramine T method. Free and bound antigen were separated using a second antibody directed against rabbit-IgG. Serum concentrations of 7 S collagen ranged from 5-14 micrograms/l. Serum levels of 7 S collagen were increased in diabetic patients as compared to normal subjects.

Adolescent↗

Presence of connective tissue proteins on the endothelium of the rat aorta.

The microdistribution of type V collagen, fibronectin, and laminin on the luminal surface of perfusion-fixed normal rat aortic endothelium has been studied by an immunoelectron microscopic method using monospecific antibodies and a protein A-gold complex. Gold particles indicating the presence of these biologically active connective tissue proteins were localized in groups on and in the vicinity of the interendothelial border. They were also found on the small flaps of cell junctions as well as on certain cell projections and scattered on the cell surface. Correlative transmission electron-microscopic examinations proved the specificity of these localizations. The endothelial cells of the aorta differed markedly in the amount of scattered connective tissue proteins on their surface, suggesting that there are several types of aortic endothelial cells with distinct functional differences. The findings provide evidence that connective tissue proteins may contribute to the surface pattern of the normal endothelium, especially on cell borders. It is likely that these proteins influence functions such as the permeability and chemotactic activity of the endothelium pertinent to the development of vascular disease.

Animals↗

New cytomorphologic methods in the diagnosis of bone tumors: possibilities and limitations.

The value of new morphologic methods in the diagnosis of bone tumors is demonstrated in a number of cases. In round cell malignancies (Ewing's sarcoma, malignant lymphoma, neuroblastoma, and anaplastic plasmacytoma) diagnostic accuracy can be improved by electron microscopic and immunohistochemical techniques. New methods are also of value in differentiating the metastatic carcinoma from malignant bone primaries. Electron microscopy may show epithelial cell features (ie, gland structures, desmosomes, and tonofilaments), while immunohistologic investigation of the cytoskeleton may facilitate differentiation of epithelial cells (positive for prekeratin) from mesenchymal cells (positive for vimentin). In the differential diagnosis of typical bone tumors, however, such as osteosarcoma, chondrosarcoma, and malignant fibrous histiocytoma, the value of enzyme histochemical, electron microscopic, and immunohistochemical methods appears somewhat restricted: alkaline phosphatase activity may be increased in both chondrosarcoma and osteosarcoma; collagen type II, the cartilage-specific collagen, is found not only in chondrosarcoma but in osteosarcoma as well. Moreover, osteosarcomas may contain a considerable number of macrophages and histiocytes, and so this feature is worthless in distinguishing osteosarcoma from malignant fibrous histiocytoma. A new approach for appraising the malignancy of bone tumors may be through flow cytometric investigation of nuclear DNA content. Osteosarcomas reveal DNA aneuploidies in more than 80% of cases, with a large proportion of cells in the S phase. These features may prove valuable for discerning osteosarcoma from myositis ossificans. In contrast to typical giant cell tumor of bone, a rare case of malignant giant cell tumor showed aneuploid cell lines indicating the malignant nature of the tumor.

Bone Neoplasms↗

Drug intoxication: influence of hemoperfusion on human plasma fibronectin.

Fibronectin is a glycoprotein belonging to the opsonic system and able to mediate phagocytosis by the reticuloendothelial system (RES). Severe intoxications are often followed by a diminished capacity of RES-clearance. Patients with severe drug intoxications have lower concentrations of fibronectin in plasma than healthy persons. Charcoal hemoperfusion lowers plasma fibronectin by an average of 4.7 mg/dl. Initial values below 15 mg/dl did not undergo a significant decrease in the case of hemoperfusion. Regarding these findings, determination of plasma fibronectin seems a useful parameter for monitoring RES function during treatment of drug-intoxicated patients by hemoperfusion.

Adolescent↗

Biologic characterization of human bone tumors. II. Distribution of different collagen types in osteosarcoma--a combined histologic, immunofluorescence and electron microscopic study.

Sixteen cases of typical highly malignant osteosarcoma were investigated by light, electron, and immunofluorescence microscopy to demonstrate the presence of collagen types I-III. It was shown that, in light-microscopically anaplastic areas of the tumor, collagen type III predominates, while only very few membranes of collagen type I are observed. Ultrastructurally, the cells are characterized by numerous free ribosomes in their cytoplasm and only a few membranes of granular endoplasmic reticulum (ER). In osteoblastic areas, collagen type I is increased, while type-III collagen is decreased. The cytoplasm of cells contains markedly more granular ER. An increasing mineralization of matrix is observed. In fibroblastic areas of the tumors, collagen types I and III are codistributed. Tumor cells have a fibroblast appearance with elongated nuclei and well developed granular ER. The chondroblastic areas, characterized by immature neoplastic cartilage, contain varying amounts of collagen type II. Chondroblast-like tumor cells have typical ring-shaped membranes of granular ER in their cytoplasm. The evidence of different collagen types in osteosarcomas lends additional support to the concept that a pluripotent mesenchymal cell is the stem cell of osteosarcomas.

Adolescent↗

Connective tissue components of the normal and fibrotic human liver. II. Clinical aspects.

In this second part, clinical aspects of connective tissue metabolism in the liver will be described and two main aspects considered. The first is the possibility to monitor the activity of fibrosis by the use of metabolites and enzymes of connective tissue metabolism. In recent studies the qualification for this purpose of enzymes such as procollagen prolyl hydroxylase and lysosomal N-Acetyl-beta-glucosaminidase and the N-terminal peptide of procollagen type III has been tested. The serum activities or concentrations of these substances in patients with chronic active liver diseases increase in due relation to the histologically estimated activity of liver fibrosis. The second aspect deals with therapeutic approaches to fibrosis in the liver by using connective tissue specific agents. So far none of the antifibrotic substances such as proline analogues, colchicine, lathyrogens and penicillamine has been used in longer-term antifibrotic treatment.

Acetylglucosaminidase↗

Biologic characterization of human bone tumors. I. Ewing's sarcoma. A comparative electron and immunofluorescence microscopic study.

Six cases of Ewing's sarcoma were investigated by electron and immunofluorescence microscopy. A layer of basement membrane-like deposits was found between typical principal and secondary tumor cells. To clarify the nature of these ultrastructural deposits, antibodies against collagen type IV were applied to frozen sections of corresponding tumor tissue. This reaction revealed type IV collagen as a regular component of basement membranes in nonneoplastic tumor capillaries, but it was equally able to localize collagen type IV between single tumor cells in capillary-free areas. With the same method, factor-VIII-associated protein, predominantly found in endothelial cells, could be demonstrated in some tumor cells. These results demonstrate that, in addition to anaplastic cells, some tumor cells are found in Ewing's sarcoma that share certain differentiating features with the endothelial cell.

Adolescent↗