The identification of four protein kinase C isoforms in human glioblastoma cell lines: PKC alpha, gamma, epsilon, and zeta.

Levels of protein kinase C (PKC) isoforms in eight human glioblastoma cell lines and two normal human glial cell cultures were determined. Earlier studies identified PKC-alpha and PKC-gamma in these cell lines but PKC-beta was not present. In this study, PKC-epsilon and PKC-zeta are demonstrated immunologically in these cell lines and also in two normal human glial cell cultures.

A search for gli expression in tumors of the central nervous system.

The gli gene was originally isolated from DNA amplified in double minutes in a glioblastoma (D259MG). Using a sensitive RNA-RNA hybridization, we tested a series of central nervous system tumors for expression of the gli gene. These included 8 glioblastoma cell lines, plus cell cultures of 5 glioblastomas, 4 anaplastic astrocytomas, 3 different ependymal tumors, a malignant meningioma, and a medulloblastoma.

Antigenic and differentiative heterogeneity among human glioblastomas.

Increasing evidence suggests that in mammals, astrocytes are a heterogenous family of cells all of which share certain properties, but differ in lineage, biochemical and functional aspects. It seems likely that glioblastomas, arising from glial precursors, may also represent a family of related but distinct cell types. We have examined the antigenic characteristics and differentiative potential of 7 different human glioblastoma cell lines in vitro.

Transcriptional patterns of growth factors and proto-oncogenes in human glioblastomas and normal glial cells.

To document over-expression of proto-oncogenes in tumors, it is necessary to determine the level of expression in the progenitor normal tissue. These studies compare the levels of nuclear transcription of a series of growth-factor related genes and proto-oncogenes in human glioblastoma cell lines with those in three normal glial cell populations. The unusual finding was that levels in the three normal glial cell populations varied considerably for several genes and thus overexpression of a specific gene in a tumor cell when compared to just one normal glial cell population would not necessarily represent overexpression.

Protein kinase C isoforms in human glioblastoma cells.

Protein kinase C (PKC), an enzyme involved in signal transduction, responds to diacyl glycerol and also to phorbol ester, a ligand analogous to diacyl glycerol. We have studied the expression of the major isoforms (alpha, beta I, beta II, and gamma) in eight human glioblastoma cell lines. In all eight lines, PKC-alpha mRNA and protein were expressed.

An epoxygenase metabolite of arachidonic acid mediates angiotensin II-induced rises in cytosolic calcium in rabbit proximal tubule epithelial cells.

Previous studies from this and other laboratories have shown that angiotensin II (AII) induces [Ca2+]i transients in proximal tubular epithelium independent of phospholipase C. AII also stimulates formation of 5,6-epoxyeicosatrienoic acid (5,6-EET) from arachidonic acid by a cytochrome P450 epoxygenase and decreases Na+ transport in the same concentration range. Because 5,6-EET mimics AII with regard to Na+ transport, it effects on calcium mobilization were evaluated.

Expression of platelet-derived growth factors, transforming growth factors, and the ros gene in a variety of primary human brain tumors.

Ribonucleic acid was isolated from a wide spectrum of central nervous system tumors to examine the expression of platelet-derived growth factors (PDGF) A and B, tumor growth factors (TGF-beta) 1 and 2, and ros messenger ribonucleic acid. Eight glioblastoma cell lines were examined as well as cell cultures from 22 tumor explants. The explants included 6 glioblastomas, 4 anaplastic astrocytomas, 5 astrocytomas, 3 ependymal tumors, 2 meningiomas, 1 medulloblastoma.

The cell cycle dependence of c-sis gene expression: artifactual conclusions in cells prepared by chemical but not physical techniques.

The c-sis oncogene encoding the B-chain of platelet-derived growth factor (PDGF) may be involved in an autocrine growth stimulation of tumours expressing the PDGF receptor, such as glioblastomas and sarcomas. To investigate whether expression of c-sis RNA is regulated in a cell cycle dependent manner, human A172 glioblastoma cells were synchronized by either centrifugal elutriation or chemical blockage with the DNA synthesis inhibitors hydroxyurea or aphidicolin. In non-perturbed elutriated cells, c-sis RNA levels were lower in the S phase of the cell cycle than in the G1 phase.

Induction of transcription from the long terminal repeat of Moloney murine sarcoma provirus by UV-irradiation, x-irradiation, and phorbol ester.

The long terminal repeat (LTR) of Moloney murine sarcoma virus (Mo-MuSV) was used as a model system to study the stress response of mammalian cells to physical carcinogens. The chloramphenicol acetyltransferase (CAT) gene was inserted between two Mo-MuSV LTRs, and the LTR-CAT-LTR construct was used for virus production and was integrated into the genome of NIH 3T3 cells in the proviral form. This construct was used to assure that the integrated CAT gene was driven by the promoter of the LTR.

Cardiovascular effects of high-frequency oscillatory and jet ventilation.

To determine the effects of mean airway pressure on hemodynamics during high-frequency ventilation, we ventilated five cats (wt 2.8 +/- 0.6 kg) using both HFOV (frequencies 3 to 20 Hz) and HFJV (frequencies 4 to 8 Hz) at Paw values ranging from 2 to 12 cm H2O.

Major structural alterations of the c-sis gene are not observed in a series of tumors of the human central nervous system.

Expression of the c-sis oncogene, the gene encoding the B chain of platelet-derived growth factor (PDGF), may be related to initiation and/or progression of glial cell tumorigenesis by PDGF-mediated autocrine growth stimulation. As the mechanism for activation of expression of the c-sis gene in gliomas is not known, we searched for possible structural alterations of c-sis DNA in these tumors. Genomic Southern blots of DNA from 7 different cultured human glioblastoma cell lines and 15 different solid human brain tumors revealed no significant change in either the gross structure or the copy number of the c-sis gene in tumor cells vs.

Control of the expression of c-sis mRNA in human glioblastoma cells by phorbol ester and transforming growth factor beta 1.

The regulation of c-sis oncogene expression in human glioblastoma cell line A172 has been investigated using a sensitive RNA-RNA solution hybridization method. Enhanced expression of c-sis mRNA was induced by phorbol ester (PMA) and diacylglycerol, each of which activates protein kinase C. c-sis mRNA was also induced by transforming growth factor beta (TGF-beta).

Expression and stability of c-sis mRNA in human glioblastoma cells.

The production of platelet-derived growth factor like (PDGF-like) material by glioblastomas may be involved in the conversion of normal cells to tumor cells. In an investigation of this problem, we have examined some of the properties of the platelet-derived growth factor B-chain mRNA (c-sis mRNA) by a sensitive and quantitative RNA-RNA solution hybridization method. In 5 out of 8 human glioblastoma cell lines, c-sis mRNA was present, and in the line with the highest level, there were approximately 4-10 molecules per cell.

Identification of Alu transposition in human lung carcinoma cells.

We have demonstrated genetic transposition in human cells. An experimental system was established in which the Ecogpt (gpt) gene was employed as a target for inactivation. The human lung carcinoma cell line A549 containing this target was fused to UV-irradiated A549 cells that did not contain the target.

Impairment of hemodynamics with increasing mean airway pressure during high-frequency oscillatory ventilation.

We investigated the effects of changes in mean airway pressure (Paw), oscillatory frequency and lung compliance on cardiac output (CO) and pulmonary vascular resistance in seven adult cats (3.0 +/- 0.6 kg) during high-frequency oscillatory ventilation (HFOV).

Effect of sleep state on chest wall movements and gas exchange in infants with resolving bronchopulmonary dysplasia.

During active sleep, neonates exhibit asynchronous chest wall movements, which have been associated with a small but significant decrease in oxygenation. To determine the effects of maturation and residual chronic lung disease on both these phenomena, we studied 11 preterm infants with resolving bronchopulmonary dysplasia (BPD) and compared these infants to ten healthy preterm infants all at time of discharge. Synchrony of chest wall (upper rib cage and abdominal) movements, sleep state, O2 saturation, and transcutaneous CO2 (TcPCO2) were recorded during both active (AS) and quiet sleep (QS).

Carbon dioxide elimination during high-frequency jet ventilation.

The effects of frequency, tidal volume, and inadvertent positive end-expiratory pressure (PEEP) on CO2 elimination were studied in rabbits during high-frequency jet ventilation by measuring CO2 concentration of expired gas using a mass spectrometer. Increasing tidal volume augmented CO2 elimination (P less than 0.01 to 0.

The effect of preincubation of HeLa cell nuclei with ATP on the degradation of mononucleosomal DNA by micrococcal nuclease.

HeLa cell nuclei with DNA labeled with [3H] thymidine have been preincubated under varying conditions and then incubated with micrococcal nuclease. Aliquots, removed at increasing times, were analyzed for mononucleosomal size DNA and for acid-soluble DNA, the ratios were plotted and a slope was determined. Preincubation with ATP and a regenerating system increased the slope 2 fold.

Release of 3-methyladenine from linker and core DNA of chromatin by a purified DNA glycosylase.

Oligonucleosomes were isolated from [14C]thymidine-labeled HeLa cells by digestion of the nuclei with micrococcal nuclease and were then alkylated with [3H]methylnitrosourea. Nucleosome core particles were also prepared by further digestion of the oligonucleosomes. The distribution of 3H-labeled methyl groups in the linker versus the core DNA was established by a determination of 3H:14C ratios in oligonucleosome and core DNA.

The release of 3-methyladenine from nucleosomal DNA by a 3-methyladenine DNA glycosylase.

Nucleosomes from chicken erythrocytes, with DNA containing an average of 144 base pairs, were alkylated with [3H]methylnitrosourea. The level of alkylation of the nucleosomal DNA was 48% of that of free DNA. The histones had approximately one tenth the radioactivity of the DNA.

Two DNA glycosylases in Escherichia coli which release primarily 3-methyladenine.

Two enzymes have been partially purified from Escherichia coli and designated 3-methyladenine DNA glycosylases I and II. The glycosylase I is that described by Riazuddin & Lindahl [Riazuddin, S., & Lindahl, T.