Clinical implications of the p53 tumor-suppressor gene.
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Antibodies, Neoplasm↗
Biomedical subjects
M Müller
Publications and source records attributed to M Müller.
4,5-Dianilinophthalimide: a protein-tyrosine kinase inhibitor with selectivity for the epidermal growth factor receptor signal transduction pathway and potent in vivo antitumor activity.
Deregulated signal transduction via the epidermal growth factor receptor (EGF-R) family of protein-tyrosine kinase growth factor receptors is associated with proliferative diseases. We describe a class of compounds (4,5-dianilinophthalimides) that inhibit the EGF-R protein-tyrosine kinase in vitro with high selectivity. In cells, 4,5-dianilinophthalmide selectively inhibited both ligand-induced EGF-R and p185c-erbB2 autophosphorylation and c-fos mRNA induction. Antitumor activity could be demonstrated in vivo against xenografts of the A431 and SK-OV-3 tumors, which overexpress the EGF-R and p185c-erbB2, respectively. In contrast, a platelet-derived growth factor-driven tumor was not inhibited by 4,5-dianilinophthalimide, which is compatible with its cellular selectivity and hypothesized mechanism of action. No overt cumulative toxicity was observed during treatment even though high efficacy was observed, indicating a good therapeutic window. 4,5-Dianilinophthalimides may offer therapeutic agents for the treatment of hyperproliferative diseases that overexpress EGF-R family protein-tyrosine kinases or their ligands.
3T3 Cells↗
pH-dependent formation of ethylenediaminetetraacetic acid supramolecular aggregates.
Pure ethylenediaminetetraacetic acid tetrasodium salt (EDTA.Na4) in water showed a pH-dependent change of optical density detected in the ultraviolet region. The most important change was found between pH 4 and 8, corresponding to the deprotonation of the third ionizable group having a pK of 6.2. The absorbance change was reversible and depended on the addition/removal of only 1 proton equivalent. The presence of supramolecular aggregates was suggested by size-exclusion chromatography, dialysis and cryo-electronmicroscopy. The molecular weight of EDTA supramolecular aggregates was 10-15,000 and 8-14,000, as determined by gel chromatography and dialysis, respectively. Cryo-electronmicroscopy revealed structures of 2-4 nm in diameter.
Chromatography, Gel↗
Selective degeneration of CA1 pyramidal cells by chronic application of bismuth.
The heavy metal bismuth induces a new type of selective neuronal degeneration that shares some common aspects with that seen following hypoxia and ischemia. Continuous application of 3 microns bismuth to organotypic cultures of rat hippocampus resulted after 2-3 weeks in selective degeneration of CA1 pyramidal cells, while CA3 pyramidal cells, dentate granule cells, and subicular neurons were resistant. With 10 microns bismuth, the majority of hippocampal neurons degenerated. The addition of 20 microns MK-801, a noncompetitive NMDA-antagonist, during the entire culturing period failed to prevent neuronal degeneration induced by 3 microns bismuth. GABA-immunoreactive interneurons were also affected by bismuth, but were generally less sensitive than CA1 pyramidal cells. Acute application of up to 100 microns bismuth did not change the electrophysiological properties of CA1 pyramidal cells.
Animals↗
Rice tungro bacilliform virus: transcription and translation in protoplasts.
Protoplasts from cell suspension cultures of Oryza sativa (monocot) and Orychophragmus violaceus (dicot) support transcription from the rice tungro bacilliform virus (RTBV) promoter and translation of the resulting mRNA despite the presence of a long leader sequence with strong secondary structure and 12 short open reading frames. Transcriptional elements located both upstream and downstream of the transcription initiation site are defined by deletion analysis and the functional TATA motif is determined. Expression of an open reading frame downstream of the entire leader is more efficient than that downstream of truncated derivatives. For optimal expression sequences in the 5' and 3' parts of the leader are required.
Base Sequence↗
Membrane assembly in bacteria.
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Bacteria↗
Immunohistological detection of the beta subunit of prolyl 4-hydroxylase in rat and mini pig lungs with radiation-induced pulmonary fibrosis.
Polyclonal and monoclonal antibodies to the beta subunit of prolyl 4-hydroxylase, the protein disulphide isomerase, were used to compare the pulmonary cells in 13 normal and in 20 fibrotic rat and mini-pig lungs made fibrotic by X-ray irradiation, using the ABC immunoperoxidase technique. In normal lungs, prominent staining of Clara cells and type II pneumocytes and weaker reactivity with alveolar macrophages, fibroblasts, endothelial and smooth muscle cells were detectable. In pulmonary disease, in which interstitial fibrosis was the characteristic feature, the immunoreactivity was increased in both the epithelial and interstitial cells. Type I pneumocytes remained negative. In the early stages of disease (3 to 4 weeks after irradiation) when little morphological alteration was seen, capillary endothelial cells had already become immunoreactive. These results underline the complex involvement and interaction of different lung cell populations in the process of pulmonary fibrogenesis.
Animals↗
Comparative long-term experience with immunoadsorption and dextran sulfate cellulose adsorption for extracorporeal elimination of low-density lipoproteins.
Two low-density lipoprotein (LDL) apheresis methods allowing a specific extracorporeal removal of atherogenic lipoproteins from plasma were compared concerning their efficacy and safety in the long-term therapy of severe familial hypercholesterolemia. Five patients were treated with immunoadsorption (IMA) at weekly intervals over 3 years each, and three patients received weekly therapy with dextran sulfate cellulose adsorption (DSA) for up to 2 years. The mean plasma volume processed per session to decrease total cholesterol to a target level of 100-150 mg/dl at the end of LDL apheresis was significantly lower in DSA than in IMA: 143% vs. 180% of the individual plasma volume. Both LDL apheresis procedures achieved a mean acute reduction of plasma LDL cholesterol by more than 70%. The average interval concentrations of plasma LDL cholesterol obtained without concomitant lipid-lowering medication were 151 +/- 26 mg/dl compared to 351 +/- 65 mg/dl at baseline in the IMA-treated patients and 139 +/- 18 mg/dl compared to 359 +/- 48 mg/dl at baseline in the DSA-treated patients. Two patients from the DSA group died after 2 years of study participation due to a stroke and a sudden cardiac death several days after the last plasma therapy. Treatment-related side effects were infrequent. Long-term therapy with IMA and DSA was associated with symptomatic improvement of coronary artery disease and mobilization of tissue cholesterol deposits. Analysis of coronary angiograms after 3 years of weekly LDL apheresis with IMA revealed in five patients nearly identical atherosclerotic lesions without definite regression or progression.
Adult↗
Localization of surfactant protein A (SP-A) in alveolar macrophage subpopulations of normal and fibrotic rat lung.
The colocalization of surfactant protein A (SP-A) and the alveolar macrophage markers ED1 and RM-1, as well as various lectins of the N-acetyl-galactosamine group [Maclura pomifera lectin (MPA), Dolichos biflorus lectin (DBA), soybean agglutinin (SBA)] and of the mannose group [Canavalia ensiformis lectin (ConA), Galanthus nivalis lectin (GNA)] was studied in normal and fibrotic rat lung tissues. In normal tissue, SP-A was located preferentially in the alveolar macrophage subpopulation lacking specific binding sites for lectins of the N-acetylgalactosamine group (DBA and SBA), although 50% of MPA-binding macrophages contained SP-A. The ED1-positive cells were SP-A-negative, whereas SP-A uptake could be detected among the RM-1 immunoreactive as well as the ConA and GNA binding macrophages. In fibrotic lung tissue, however, a small number of DBA and SBA binding macrophages contained SP-A and the percentage of GNA and ConA binding alveolar macrophages exhibiting SP-A immunoreactivity was reduced. Additionally, the number of ED1+/SP-A+ macrophages was found to be increased. Immunoelectron microscopy revealed accumulation of SP-A in the extracellular space. The differing SP-A content in different alveolar macrophage subpopulations suggests a more complex mechanism of uptake and degradation of surfactant proteins in normal and pathological conditions, which cannot simply be explained by the glycoconjugate pattern on the surface of alveolar macrophages.
Animals↗
Immunohistochemical detection of tissue factor (TF) on paraffin sections of routinely fixed human tissue.
Tissue factor (TF), a 47 kDa transmembrane glycoprotein, is the essential receptor and cofactor for factor VII/VIIa. Its distribution in normal tissues and in tumours has been recently investigated immunohistochemically with monoclonal and polyclonal anti-TF antibodies in frozen sections. The cardinal problem of this technique is the difficulty of determining exactly the localization of the reaction product at least in certain tissues. Here, we demonstrate a method using monoclonal anti-TF antibodies to detect TF in routinely fixed, microwaved, paraffin-embedded tissues. Generally, there were no fundamental differences in TF distribution in frozen and paraffin-embedded material. However, in most cases, the paraffin sections allow a better cellular localization of TF. For example, the staining pattern for TF in both kinds of sections is identical in kidney, brain and skin. The paraffin-embedded material, however, clearly shows that TF expression is restricted to the parietal and the visceral epithelia of Bowman's capsule of glomeruli in the kidney, and to astrocytes and their processes in the brain. TF reactivity in the skin is revealed to be cell membrane-bound; in cardiomyocytes TF shows an exclusively sarcolemmal localization. The immunohistological detection of TF in paraffin sections is a powerful tool for systematic studies on the possible role of TF in the context of physiological and pathological studies.
Antibodies, Monoclonal↗
Ultrastructure of hypertrophic cartilage: histochemical procedures compared with high pressure freezing and freeze substitution.
The effect of cationic dyes on the ultrastructure of hypertrophic cartilage was compared with results obtained with modern cryotechniques in studies on rat epiphyseal growth plate. Addition of alcian blue, acridine orange, cupromeronic blue, ruthenium hexamine trichloride, ruthenium red, or safranin O to conventional glutaraldehyde/osmium tetroxide fixatives to a large extent resulted in prevention of chondrocyte shrinkage except for alcian blue which showed poor tissue penetration. The fine structure of the matrix in pericellular and territorial compartments appeared very coarse with areas of high contrast in tissue exposed to fixatives containing cationic dyes. This indicates structural collapse and precipitation of electron-dense material, a pattern clearly differing from that observed in specimens prepared by the cryotechniques. The dyes giving a pattern most similar to that seen after high pressure freezing, freeze substitution, and low temperature embedding were acridine orange and safranin O. It is concluded that studies of matrix ultrastructure down to the molecular level necessitate the application of cryotechniques.
Animals↗
Colocalization of cytokeratin 18 and villin in type III alveolar cells (brush cells) of the rat lung.
Alveoli of the rat lung are lined by three different cell types, the flat type I cells and the cuboidal type II and type III cells. Type III cells differ from type II cells by the presence of an apical tuft of microvilli and the absence of lamellar type secretory granules. In the present study we show by double immunolabelling that type III cells of the rat lung can be identified at the light- and electron microscope level by antibodies against both cytokeratin 18 and the actin-crosslinking protein villin. At the ultrastructural level, microvilli and their rootlets in the apical cytoplasm were labelled by the anti-villin antibodies, whereas a monoclonal antibody against cytokeratin 18 (Ks18.04) labelled bundles of intermediate filaments. In conclusion, antibodies against villin and certain monoclonal antibodies specific for cytokeratin 18 can be used as tools for selective visualization of type III cells in the rat lung.
Animals↗
The influence of methylprednisolone on the energy metabolism of Ehrlich ascites tumour cells.
Using Ehrlich ascites tumour cells, the short-term effects of the therapeutic glucocorticoid Methylprednisolone (MP) on the cellular energy metabolism were studied. ATP-consuming processes involved in the rapid MP effects were identified indirectly from the effects of MP on cellular oxygen consumption related to the inhibition of respiration by selective inhibitors of Ca(2+)-ATPase and protein synthesis. The effects of MP on plasma membrane permeability for Ca2+ ions and phospholipid turnover were studied directly by using confocal laser scanning microscopy and tracerkinetic measurements, respectively. MP inhibited cellular oxygen consumption, suppressed the inhibitory effect of lanthanum but not that of cycloheximide on oxygen consumption, blocked the [Ca2+]i rise in response to calcium ionophore A 23187, and decreased phospholipid turnover. MP acted instantly in a dose-dependent manner. The observed effects of MP are discussed in relation to the hypothesis that the drug has direct membrane effect affecting plasma membrane permeability and function.
Adenosine Triphosphate↗
Antibodies against linear and conformational epitopes of the human papillomavirus (HPV) type 16 E6 and E7 oncoproteins in sera of cervical cancer patients.
Sera obtained from 137 cervical cancer patients were analysed for the presence of antibodies to the human papillomavirus (HPV) type 16 proteins E6 and E7 by the aid of different assays, i.e. ELISA using as antigen either synthetic peptides or the complete E7 protein and radio-immunoprecipitation (RIPA) which uses the viral protein made by in vitro transcription/translation. In agreement with previous reports, reactivity to the E7 protein was found more frequently than to the E6 protein (31.4% vs. 16.8%) when the sera were assayed by peptide-based ELISA. In contrast, when RIPA was employed, reactivity to either protein was obtained at similar frequency (38.7% vs 46.7%). When the protein was denatured prior to immuno-precipitation the reactivity was lost in all sera tested for E6-specific antibodies but only in a few samples in the E7-RIPA. Therefore it was concluded that the increased sensitivity of the E6-RIPA as compared to the E6 peptide-ELISA is due to the detection of antibodies to conformational epitopes which are presented by the in vitro product but not by the synthetic peptides. Eighty-two sera from healthy donors were tested by HPV 16E6- and E7-RIPA and also by ELISA using the HPV 16E7 protein which was produced in the fission yeast Schizosaccharomyces pombe. One sample reacted each in the E6- and E7-RIPA indicating a high specificity of these assays. The E7 protein-ELISA proved to be less sensitive for the detection of antibodies in cervical cancer patients' sera (22.6% positive) as compared to peptide-based ELISA or RIPA.
Adolescent↗
Morphometric study of centrilobular vessels in the rat liver after continuous hypothermic perfusion with Euro-Collins and University of Wisconsin solution.
Damage to the hepatic microcirculation plays an important part in the preservation-related loss of graft viability. In a study on the isolated, hypothermically preserved rat liver perfused continuously with Euro-Collins and University of Wisconsin solution (gluconate substituted for lactobionate) at different flow rates we determined changes in diameter of central veins and pericentral sinusoids and related them to weight changes. While post-perfusion liver weight increased with Euro-Collins and decreased with University of Wisconsin solution, weight changes did not vary significantly with different flow rates. Changes in sinusoidal diameter depended on the type of solution as well as on the flow rate. Cell swelling under Euro-Collins perfusion leads to compression of pericentral sinusoids, which varies in degree with different flow rates. Perfusion with University of Wisconsin solution under near-normal flow, due to slight shrinking of parenchymal cells, keeps the sinusoidal lumina wide open. We conclude that there is a causal relationship between perfusion-dependent cell swelling and reduction in sectional area of the hepatic microvasculature. Continuous hypothermic perfusion with University of Wisconsin solution at physiologic flow rates excellently preserves the size and trabecular architecture of hepatocytes and thereby the sinusoidal lumina within the hepatic lobules.
Adenosine↗
Cholestasis caused by inhibition of the adenosine triphosphate-dependent bile salt transport in rat liver.
BACKGROUND/AIMS: Inhibition of bile salt transport across the hepatocyte during cholestasis induced by cyclosporin A has been shown. However, the contribution of the different bile salt transport systems in liver to cholestasis has remained controversial. METHODS: The sensitivity of different bile salt transport systems in liver to cyclosporin-induced inhibition was determined by transport assays in plasma membrane vesicles and by in vivo studies in the rat. RESULTS: Cyclosporin A--induced inhibition of sodium-dependent uptake of bile salts across the sinusoidal membrane, of potential-dependent, and of adenosine triphosphate (ATP)-dependent bile salt transport across the canalicular membrane exhibited inhibition constants (Ki) of 5, 70, and 0.2 mumol/L, respectively. The nonimmunosuppressive cyclosporin analogue PSC 833 also preferentially inhibited the ATP-dependent bile salt transport with an inhibition constant of 0.6 mumol/L. Cyclosporin A and its analogue PSC 833 [(3'-oxo-4-butenyl-4-methyl-Thr1)-(Val2)-cyclosporin] (25 mg/kg each) served as tools to interfere with [14C]taurocholate secretion into bile in vivo, causing an accumulation of [14C]-taurocholate in liver and reducing bile flow to 50%. In mutant rats deficient in the transport of leukotriene C4 and related conjugates across the canalicular membrane, bile flow was reduced to 14%. CONCLUSIONS: The cyclosporins preferentially inhibit the ATP-dependent bile salt export carrier in the canalicular membrane. This inhibition reduces bile salt-dependent bile flow and causes intrahepatic cholestasis.
Adenosine Triphosphate↗
Primary structure of the hydrogenosomal adenylate kinase of Trichomonas vaginalis and its phylogenetic relationships.
Hydrogenosomal adenylate kinase of the amitochondriate protist, Trichomonas vaginalis, has been purified and the sequence of its 39 amino-terminal residues established. Based on this sequence and a conserved internal region of the enzyme, a probe was obtained by DNA polymerase chain reaction and used to isolate a genomic DNA clone containing the gene of this enzyme. This gene exists probably as a single copy in T. vaginalis and is not interrupted by introns. The open reading frame obtained codes for a large type adenylate kinase with a mature molecular mass of 24.5 kDa. The T. vaginalis enzyme is homologous with adenylate kinases of other eukaryotes and eubacteria. Strongly conserved parts and residues of the molecule are conserved also in this enzyme. Phylogenetic trees obtained with various methods placed the T. vaginalis adenylate kinase close to the point where the different subfamilies of this enzyme branch from each other, indicating that the T. vaginalis enzyme has no close relationship to any of these subfamilies and that it separated early from other adenylate kinases. The conceptual translation predicts the existence of an amino-terminal nonapeptide absent from the protein purified from hydrogenosomes, similar to the processed amino-terminal extensions of other hydrogenosomal proteins. These extensions have been considered as putative targeting and import signals.
Adenylate Kinase↗
A comparative assessment of cerebral haemodynamics in the basilar artery and carotid territory by transcranial Doppler sonography in normal subjects.
In 69 healthy volunteers (34 males, 35 females, age range 17-80 years) we compared the following haemodynamic parameters between the basilar artery and carotid artery system, assessed by transcranial Doppler sonography: mean blood velocity, pulsatility index and the hemispheric indices as ratios of the middle, anterior and posterior cerebral arteries with the internal carotid artery (MCA/ICA, ACA/ICA, PCA/ICA) and of the PCA with the basilar artery (PCA/BA), as well as the ACA/MCA and the MCA/BA ratio. In all arteries (ICA, MCA, ACA, PCA and BA) mean blood velocity decreased significantly with advancing age (p < 0.01) and was significantly higher in females as compared to males (p < 0.05). The pulsatility index increased significantly with age in the ICA, MCA, ACA and BA (p < 0.01) and showed no sex differences in any of the intracranial arteries except for the ACA. The MCA/ICA and ACA/ICA index declined significantly with advancing age (p < 0.005, p < 0.05, respectively) and with increasing heart rate (p < 0.01 for both arteries), while the ACA/MCA, the PCA/BA and the MCA/BA ratio remained unchanged by age and heart rate. We conclude that there is no striking difference in the cerebral haemodynamics of the basilar artery and the carotid artery supplied territories.
Adolescent↗