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Biomedical subjects

K Weber

Publications and source records attributed to K Weber.

At least 559 records · Page 31Linked to original sources

Centriole number and process formation in established neuroblastoma cells and primary dorsal root ganglion neurones.

Electron microscopy on serial sections has been used to compare the number and position of centrioles and processes in four neuroblastoma clones and in primary dorsal root ganglion neurones. Studies of Neuro-2a and NB41, as well as further experiments on N18 and N115 confirm that a sizeable percentage of the cells in all four neuroblastoma clones have multiple centrioles (2-12 per cell). No significant differences were seen between the number and arrangement of centrioles in the undifferentiated or differentiated states. In contrast primary dorsal root ganglion neurones, although characterized by multiple processes, exhibit only two centrioles per cell, usually located as a pair on one side of the nucleus. The observation that both dorsal root ganglion neurones, as well as neuroblastoma cells with only two centrioles were capable of multiple process formation suggests that multiple centrioles are not obligatory for multiple process formation. In addition no consistent relationship was seen between the location of the centrioles and the position of the processes casting further doubts on models which assume a direct relation between these characteristics.

Animals↗

[Morphologic studies on the venous blood drainage of the human placenta].

A human in situ placenta of the eleventh week of pregnancy was totally serial sectioned and light microscopically examinated with following results: 1. 208 venous openings were counted 2. Venous drainage takes place over the whole basal plate 3. the venous openings are scattered in different regions a) mainly responsible for the venous drainage are the border parts of the placenta between central parts and the marginal lake b) the openings lie together in cricoid groups. This pleads for the existence of circulation units 4. there is no relationship perceptible between venous openings and placental septa 5. venous valves and arteriovenous anastomoses do not exist.

Chorionic Villi↗

F-actin binding and bundling properties of fimbrin, a major cytoskeletal protein of microvillus core filaments.

Fimbrin, previously recognized as a major structural protein of the microfilament core bundles of intestinal epithelial cell microvilli, has been purified to homogeneity and characterized. It is a nearly globular monomeric protein of apparent molecular weight 68,000 and has a single calcium binding site (Kd = 9 microM), for which magnesium ions compete. Fimbrin binds to F-actin and this interaction is characterized in detail. Under our optimal binding of conditions, fimbrin induces tightly packed F-actin bundles, similar to the bundles induced by villin, another microvillus structural protein. The formation of mixed fimbrin-villin-actin bundles provides a further step toward the full in vitro reconstitution of microvillus core filaments from its purified individual components. The reconstituted fimbrin-villin-actin bundles do not display the side arms characteristic of isolated microvillus cores. These results are discussed in terms of our current understanding of the organization of the microvillus core filaments and indicate that this structure contains two bundling proteins, villin and fimbrin. The results complement previous studies and now provide a minimal biochemical characterization of all four major actin-associated structural proteins so far identified in core filaments. Three of these (villin, fimbrin, and calmodulin) are calcium-binding proteins, emphasizing the concept of calcium control over submembranous microfilament organization.

Actins↗

Demonstration of at least two different actin-binding sites in villin, a calcium-regulated modulator of F-actin organization.

Villin, one of the calcium regulated modulator proteins of F-actin organization, restricts F-actin to short filaments in the presence of calcium and bundles F-actin in the absence of calcium. Limited in vitro proteolysis of villin generates, in addition to a large core fragment (apparent Mr = 90,000) previously described, a small headpiece (Mr = 8,500). The finding that the F-actin nucleation and severing activity of villin, but not its bundling activity, is retained by the core suggested that the headpiece may be directly involved in bundling. Headpiece has now been purified and characterized. It shows strong F-actin binding both in the presence and absence of calcium, leading to a final stoichiometry of 1 headpiece to 1 F-actin monomer. Headpiece also inhibits villin-induced F-actin bundling. Thus villin expresses at least two distinct actin-binding sites localized on separate functional domains. Protein sequence analysis documents that the core comprises the NH2-terminal portion of intact villin, whereas the headpiece covers the COOH-terminal 76 amino acids. We provide the amino acid sequence of the headpiece, which is currently the smallest F-actin binding peptide.

Actins↗

Centrioles, microtubules and microfilaments in activated mononuclear and multinucleate macrophages from rat peritoneum: electron-microscopic and immunofluorescence microscopic studies.

In male Wistar rats of the BD I strain, mononuclear macrophages and multinucleate giant cells obtained from the peritoneum 1 day to 5 weeks after implantation of coverslips coated with dermoid cyst sebum, were examined by light microscopy and immunofluorescence microscopy, using antibodies specific for actin and tubulin and also by scanning and transmission electron microscopy. In activated mononuclear macrophages, microtubules radiate from the centrioles, situated in the perinuclear area, into the cytoplasm and the major cell processes. Microfilaments form a dense meshwork beneath the plasmalemma. When mononuclear macrophages fuse to form multinucleate giant cells, the initially unordered ("Foreign body") syncytia still reveal the original distribution patterns of centrioles, microtubules and microfilaments similar to those seen in the individual cells. In the ordered (Langhans) multinucleate giant cell all centrioles are accumulated in a main pluricorpuscular central group. Centrioles are the initiating and organising centres in the formation of microtubules. From the centrioles microtubules extend into the entire cytoplasm of the syncytium as a uniformly organised, stellate, radial system. The centrosphere, which is characteristic for ordered multinucleate giant cells, seems free from microfilaments, which form a ring-shaped woven network encircling the nuclei. Depolymerisation and inhibition of microtubules upon exposure to colchicine, indicates that both the organisation of the cytoplasm and the cellular movements depend on the undisturbed coordination of centrioles, microtubules and microfilaments. This applies also to the fusion of mononuclear macrophages to form syncytia, the ordering processes within multinucleate giant cells, and the function of ordered giant cells.

Animals↗

The distribution of keratin type intermediate filaments in human breast cancer. An immunohistological study.

Antibodies to different intermediate filament proteins can be used to distinguish cells of epithelial, mesenchymal, muscle, glial and neuronal origin. Antibodies to prekeratin which characterize cells of epithelial origin, and antibodies to vimentin which recognize cells of mesenchymal origin have been used to study twenty cases of breast carcinoma (sixteen infiltrating ductal carcinomas and four infiltrating intraductal carcinomas), two cases of cystic breast disease, two fibroadenomas and one case of benign cystosarcoma phylloides. The prekeratin and vimentin were detected using specific antibodies to these proteins by immunofluorescence microscopy using alcohol fixed paraffin-embedded tissues. In eighteen out of the twenty carcinomas the tumor cells were strongly and specifically stained by antibodies to prekeratin. DIfferent tumors gave different patterns of prekeratin staining. In contrast, when the same specimens were tested with the vimentin antibody, the tumor cells were unstained, and instead only the usual strong staining to fibroblasts and blood vessels in the stroma was observed. In cystic breast disease, fibroadenomas, and benign cystosarcoma phylloides, cells of epithelial origin were strongly stained by the prekeratin but not by the vimentin antibody.

Adenofibroma↗

On interactions of cytostatic benzylidine hydrazines with SH-groups.

The cytostatic and mutagenic compound N'-methyl-N'-cyano-(p-chloro)-benzaldehyde hydrazone (CyB4) has been found to be a strong SH-blocking agent since it reacts with the thiol groups of glutathione and of the cell membrane of Ehrlich ascites carcinoma cells (EAC). Furthermore, it decreases the intracellular, non-proteinogenic SH(NPSH)-level of tumor cells. CyB4 is not able to alkylate 4-(p-nitrobenzyl)-pyridine (NBP) by a nucleophilic substitution reaction, but it could be shown that the reactivity of CyB4 with thiol groups is due to a Michael-addition-type reaction of SH-groups with the cyano-group of CyB4. On the other hand, cytostatic beta-chloroethyl hydrazones showed a negligible reactivity against glutathione and led even to an increase of thiol groups, detectable by the 5,5'-dithiobis-2-nitrobenzoic acid (DTNB)-method, at the cell membrane of EAC when incubated in the presence of beta-chloroethyl hydrazones N-benzylidene-N'-methyl-N'-(2-chloroethyl) hydrazine (B1) and N-(4-dimethylaminobenzylidene)-N'-methyl-N'-(2-chloroethyl) hydrazine (B2). Therefore, it is concluded that the cytostatic efficiency of CyB4 is due to its SH-blocking while that of the beta-chloroethyl hydrazones is due to a rearrangement of the tumor cell membrane, as indicated by the increased level of reactive SH-groups.

Alkylating Agents↗