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Succinyl concanavalin A stimulates and antimicrotubular drugs inhibit the synthesis of a brain-specific protein in rat glial cells.

The relative synthesis of the brain-specific s100 protein increased as clonal rat glial cells, C6, progressed from logarithmic to stationary growth in monolayer culture. Drugs that disrupt microtubules, such as colchicine, vinblastine, Colcemid, and podophyllotoxin, inhibited the relative synthesis of S100 protein in stationary cultures. Colchicine (0.1 muM) caused a 50% inhibition of the relative synthesis of S100 protein whereas lumicolchicine,an isomer of colchicine that does not disrupt the microtubular system, had no effect. Succinylated concanavalin A (500 mug/ml) increased relative synthesis in logarithmic but not stationary cultures. These results suggest that signals inducing an increase in the relative synthesis of S100 protein in stationary cultures are transmitted intracellularly from the cell membrane by the microtubular network.

Cell Line

Description of a human papillary thyroid carcinoma cell line. Morphologic study and expression of tumoral markers.

BACKGROUND: The establishment of cell lines from thyroid carcinomas can provide an in vitro model of oncogenesis. B-CPAP is a new cell line that has been obtained from a differentiated papillary thyroid carcinoma. The data presented give a broader characterization and expression of tumoral markers of this cell line and identify the differentiated functions that are preserved. METHODS: An ultrastructural study was performed to confirm the thyroid nature of the new cell line. The cellular markers (thyroglobulin, S100, neuron-specific enolase [NSE]) and the oncogenes (mutated p53, H-ras, c-myc, PTC, trk) were studied by immunohistochemistry, Southern blot, or in situ hybridization. RESULTS: The cells were of a differentiated ultrastructural thyroid type. All of the cells proved immunoreactive with antibodies specific to thyroglobulin, S100 proteins, NSE, and mutant p53 protein. Mutations of H-ras, PTC, and trk were not observed. The c-myc gene was not amplified. CONCLUSIONS: The cell line described in these data provides a suitable model for the study of thyroid carcinogenesis, given that the cells present thyroid characteristics, and metabolic disorders not previously found in such cell lines. In addition, the coexpression of S100 proteins and mutant p53 proteins in the cells should permit the study of the interaction between these two proteins.

Aged

Fate of mRNA following disaggregation of brain polysomes after administration of (+)-lysergic acid diethylamide in vivo.

Intravenous injection of (+)-lysergic acid diethylamide into young rabbits induced a transient brain-specific disaggregation of polysomes to monosomes. Investigation of the fate of mRNA revealed that brain poly(A+)mRNA was conserved. In particular, mRNA coding for brain-specific S100 protein was not degraded, nor was it released into free ribonucleoprotein particles. Following the (+)-lysergic acid diethylamide-induced disaggregation of polysomes, mRNA shifted from polysomes and accumulated on monosomes. Formation of a blocked monosome complex, which contained intact mRNA and 40-S plus 60-S ribosomal subunits but lacked nascent peptide chains, suggested that (+)-lysergic acid diethylamide inhibited brain protein synthesis at a specific stage of late initiation or early elongation.

Animals

In vitro segregation of different cell lines with neuronal and glial properties from a stem cell line of rat neurotumor RT4.

A clonal stem cell line, RT4-AC, of the rat peripheral neurotumor RT4 differentiates in culture into morphologically distinct cell types RT4-B, RT4-D, and RT4-E (cell type conversion). The multipotential stem cell type RT4-AC and cell type RT4-D produce a glial marker, S100 protein, but RT4-B and RT4-E do not. The stem cells also show a small but significant response to veratridine on voltage-dependent Na+ influx. Cell types RT4-B and RT4-E show a clear response of voltage-dependent Na+ influx to veratridine, typical of neuronal cells, whereas cell type RT4-D is completely negative. These results indicate that (i) the stem cell type RT4-AC shows both neuronal and glial properties, (ii) cell types RT4-B and RT4-E have a neuronal property, and (iii) cell type RT4-D has a glial property. Therefore, cell type conversion of stem cell RT4-AC to RT4-B and RT4-E cells seems to result in differentiation towards neuronal cell types, and cell type conversion of RT4-AC to RT4-D results in differentiation towards a glial type in culture.

Action Potentials

Glioblastoma multiforme: morphology and biology.

Glioblastoma multiforme, representing about 50% of all gliomas, encompasses a group of intrinsic tumours of the brain in later years (age peak around 50 years), the morphological hallmarks of which are an ensemble of variations in tumour cell and tissue structure featuring its biological malignancy. Glioblastoma, while sometimes appearing as a distinct "primary" tumour type, is usually accepted as an extreme manifestation of anaplasia and dedifferentiation of glia, mostly astrocytic. The astrocytic nature of most glioblastomas has been confirmed by ultrastructural studies and progressive differentiation of tumours maintained in organotypic tissue culture. Reproducible experimental models are particularly induced by oncogenic RNA (oncorna) viruses. The cell kinetic parameters are similar to those of other solid malignant tumours except for a comparatively low growth fraction of glioblastoma. The frequent occurrence of giant cells as well as of regressive changes with necrosis and vascular responses are indirect (secondary) indicators of malignancy which coincide with histochemical (enzymatic anisochronia) and biochemical data (lower level of glia specific S100 protein than in differentiated gliomas). Vascular proliferation, a characteristic feature of glioblastoma, may occasionally progress to sarcomatous transformation with development of gliosarcomas (mixed glial-mesenchymal tumours). While dissemination of glioblastoma through the cerebrospinal pathways is not uncommon, extraneural distant metastatic spread is rare, and usually observed after craniotomy. The results of modern neuro-oncology support the pathogenetic view that glioblastoma results from neoplastic transformation of glial elements with continuing dedifferentiation. This transformation can be experimentally induced by various factors including oncogenic DNA (oncorna) viruses by using a reverse transcriptase, while there is indirect evidence for an oncorna-virus information in human glioblastoma. The significance of immunological factors in the pathogenesis of brain tumours and in the course of neoplastic transformation of glia is not yet understood, but both morphological and immunological data are in favour of a cell mediated immunological reaction against tumour-specific antibodies. Since immunological factors and changes in cytokinetics are apparently active after the transformed tumour cells proliferate, all available therapeutic methods, including radiation, chemotherapy, and immunotherapy of glioblastoma only influence the final stages of neoplastic development with clinical manifestation of the tumour. In spite of modern combination and multimodality therapy schemes the prognosis of glioblastoma is still poor.

Adult

The nervous system-specific S-100 antigen in cerebrospinal fluid of multiple sclerosis patients.

The nervous system-specific S-100 antigen has been found in cerebrospinal fluid (CSF) of 13 out of 18 patients with multiple sclerosis (MS), whereas it was undetectable in either of the 11 control patients with minor psychic disturbances or with neurological disorders not usually associated with apparent parenchymal lesion. The levels of the antigen appeared to be higher in CSF of patients in the acute phase of the disease. Though the small number of cases hampers final statements, the S-100 in CSF might serve as a possible index of active cell injury in the central nervous system underlying the pathogenesis of MS.

Cerebrospinal Fluid Proteins

Antigens in human glioblastomas and meningiomas: Search for tumour and onco-foetal antigens. Estimation of S-100 and GFA protein.

Extracts of glioblastomas and meningiomas were analysed by quantitative immunoelectrophoresis for the presence of foetal brain antigens and tumour-associated antigens, and levels of 2 normal brain-specific proteins were also determined. The following antibodies were used: monospecific anti-S-100 (glia specific); monospecific anti-GFA (glial fibrillary acidic protein), (astroglia specific); polyspecific anti-foetal brain (12-16th week of gestation); a polyspecific anti-glioblastoma antiserum, absorbed with insolubilized serum, haemolysate and normal brain extract; polyspecific anti-alpha-foetoprotein; and monospecific anti-ferritin. Using the antibodies raised against the tumours, several antigens not present in foetal or adult normal brain were found in the glioblastomas and the meningiomas. These antigens cross-reacted with antigens present in normal liver and were therefore not tumour-associated. S-100 was found in glioblastomas in approximately one tenth the amount in whole brain homogenate, whereas GFA was found 2-4 times enriched. The 2 proteins were absent in meningiomas. The possible use of the GFA protein as a marker for astroglial neoplasia is discussed. Five foetal antigens were found in foetal brain, but none in the tumours. alpha-Foetoprotein could only be demonstrated in foetal tissue extracts, including foetal brain, but not in tumours. Ferritin was detected in all tumour extracts, although the amounts determined were unrelated to histological tumour type.

Antigens, Neoplasm

[Study of bromthymol blue-prealbumin in the blood serum of rats under hyperbaric oxygenation].

Three-fold increase of BTB-prealbumin (Rm 1.0) in rat serum following fierse convulsions under hyperbaric oxygenation (6 ati, 30-35 min) has been proved by disc electrophoresis. Glial S100-protein and 7-fold increase in the all-organ component of brain BTB-prealbumin were found by immunochemistry to appear in the serum of experimental rats. The consequences of disorders in the blood-brain barrier for non-specific, all-organ proteins and potentialities of protein output from the brain into the blood similarly to neurophysins under hyperbaric oxygenation are discussed.

Animals

[Several properties of one of the acid neurospecific antigens].

The identity of one of the acid neurospecific antigens (antigen A) and S-100 protein has been established through agar gel immunoelectrophoresis and Sephadex G-100 gelchromatography. On agar gel electrophoresis antigen A moves as 2 fractions which occupy positions of blood serum prealbumins and alpha2 globulins. Both fractions of the antigen have a molecular weight of 25000. The heterogenic nature of antigen A (protein S-100) during electrophoresis on agar gel is discussed.

Agar

[Heterogeneity of neurospecific protein S-100].

Protein S-100 specific for the nervous system was isolated from the bull cerebral hemispheres by the Steward method and antiserum monospecific to it was obtained. In immunoelectrophoretical study of the initial brain extract with antiserum detected the paired line of precipitation consisting of two arcs in the zone of prealbumins and alpha2-globulins of the the serum, and that with purified protein fraction showed only one arc corresponding to the prealbumin component. The purified protein S-100 fraction in agar gel is divided into a series of prealbumin electrophoretic zones, with a pre-protein locating chiefly in the most mobile zone. It is established that heterogeneity of protein S-100 with electrophoresis in agar gel and immunophoresis is due to the presence of calcium ions in the systems. In the presence of 1 mM EDTA solution the one or two closely located prealbumin zones of purified protein S-100 migrates. The obtained monospecific antiserum made it possible to establish that two-fractional antigen A detected previously by the heterogenic antisera is protein S-100.

Animals