phosphorus-radioisotopes and Carcinoma--Ehrlich-Tumor

In Part 1 of this article physical and chemical effects of beta-decay in labelled molecules were reviewed and their potential importance for breaking predetermined and specific bonds were pointed out. After incorporation of labelled biomolecules in living systems, such as viruses, phages or cells, the radioactive decay of the label alters the biological behaviour of the system, in the extreme case causing loss of the ability to reproduce, the extent of these consequences depending strongly on the type of radioisotope. Now Part 2 includes a brief discussion of biological effects associated with beta-decay emphasizing the relative importance of local transmutation and internal radiation effects from the decay of 3H, 14C, 32P, 33P, 35S and 125I. Attempt is also made, whenever possible at the present stage of understanding, to correlate biological effects with chemical processes on a molecular level.

Topics: Animals; Carbon Radioisotopes; Carcinoma, Ehrlich Tumor; Cell Nucleus; DNA; Idoxuridine; Iodine Radioisotopes; Phosphorus Radioisotopes; Radiation, Ionizing; Sulfur Radioisotopes; Tritium

Topics: Animals; Biological Transport, Active; Calcium Radioisotopes; Carbohydrates; Carcinoma, Ehrlich Tumor; Cell Membrane; Cell Membrane Permeability; Cells; Erythrocytes; Hydrogen Peroxide; Iodine Radioisotopes; Lysosomes; Mathematics; Membrane Potentials; Membranes, Artificial; Models, Biological; Phospholipids; Phosphorus Radioisotopes; Proteins; Radiation Effects; Radioisotopes; Sodium Isotopes; Water

Topics: Amino Acid Sequence; Animals; Autoradiography; Carcinoma, Ehrlich Tumor; Chromatography, Affinity; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Electrophoresis, Polyacrylamide Gel; Indicators and Reagents; Isoenzymes; Kinetics; Mice; Molecular Sequence Data; Molecular Weight; Oligopeptides; Phosphorus Radioisotopes; Phosphorylation; Protein Kinases; RNA Polymerase II; Substrate Specificity

A 32P-post-labeling method has been employed to detect DNA adducts formed by derivatives of nitro-9-aminoacridine in both cellular and non-cellular systems. The treatment of HeLa S3 cells in culture or Ehrlich ascites tumor cells in vivo with nitracrine and two other antitumor 1-nitro-9-aminoacridines, denoted C-857 and C-1006, resulted in covalent binding of these compounds to cellular DNA. Each derivative studied gave rise to a distinct pattern of adduct spots and the similarity of the respective adduct profiles was noted for the both cellular models. Calf thymus DNA samples modified in vitro with nitracrine and C-857 in the presence of either rat hepatic microsomal fraction or dithiothreitol yielded chromatographic profiles resembling those obtained in the cellular systems, suggesting similarity in the DNA adduct structures. There were also neither qualitative nor quantitative differences in calf thymus DNA modification by these two 1-nitro derivatives between aerobic and anaerobic conditions, thus the reduction of a nitro group seems not to be the only determinant of covalent binding to DNA in vitro. No DNA adduct formation was detected in the cellular systems used with 2-nitro and 4-nitro isomers of nitracrine that are devoid of cytotoxic activity, which provides further evidence that both covalent binding and DNA crosslinking, but not intercalation, are responsible for cytotoxic and antitumor properties of 1-nitro-9-aminoacridines.

Topics: Aminoacridines; Animals; Antineoplastic Agents; Carcinoma, Ehrlich Tumor; Cattle; DNA; DNA, Neoplasm; HeLa Cells; Humans; In Vitro Techniques; Isomerism; Male; Mice; Mice, Inbred BALB C; Microsomes, Liver; Nitracrine; Phosphorus Radioisotopes; Rats; Rats, Inbred Strains

Topics: Adenine Nucleotides; Animals; Buffers; Carcinoma, Ehrlich Tumor; Chromatography, Gel; Chromatography, Ion Exchange; Endoribonucleases; Interferons; Kinetics; Mice; Oligoribonucleotides; Phosphorus Radioisotopes; Protein Binding

Ehrlich ascites tumor cells were made permeable to nucleoside triphosphates by treating them with diethylaminoethyl (DEAE)-dextran. Permeable cells incorporated labeled UTP into RNA at a constant rate for at least 30 min at 37 degrees C and 100 min at 25 degrees C, whereas no incorporation was detected without DEAE-dextran treatment. This RNA synthesis is dependent on added nucleoside triphosphates and is affected differentially by different concentrations of alpha-amanitin. The results of both hybridization and S1-nuclease protection mapping experiments with pulse-labeled RNA synthesized in this system indicate that reinitiation of rRNA transcription takes place from the physiological initiation site in cells made permeable by treatment with DEAE-dextran. By using this system, we examined the kinetics of rRNA synthesis in mouse FM3A cells during inhibition of protein synthesis. The results reflected those previously obtained by analyses of nucleolar RNA synthesis in vivo during protein synthesis inhibition. This permeable cell system should provide a rapid and reproducible assay method for rRNA synthesis in mammalian cells.

Topics: Animals; Carcinoma, Ehrlich Tumor; Cell Line; Cell Membrane Permeability; DEAE-Dextran; Dextrans; Kinetics; Mammary Neoplasms, Experimental; Mice; Nucleic Acid Hybridization; Phosphorus Radioisotopes; Protein Biosynthesis; RNA, Ribosomal; Transcription, Genetic; Tritium

A phosphate-incorporating protein has been highly purified from the cytosol of Ehrlich ascites tumor cells (EAT cells). The nitrocellulose membrane method was used to follow the progress of the purification by quantitation of the [32P]phosphorylated form of the protein. The purified protein was identified as an NDP-kinase since it exhibited NDP-kinase activity and had enzyme characteristics in common with other NDP-kinases from various mammalian cells. The purified NDP-kinase was found to have a molecular weight of approximately 76,000 daltons. Moreover, the enzyme appears to consist of two distinct polypeptides (18,000 and 20,000 daltons). This enzyme contained 19 amino acids, with high levels of glycine (9.8%) and lysine (9.0%). The enzyme rapidly formed a [32P]phosphoenzyme when incubated with [gamma-32P]ATP in the presence of Mg2+ (1 mM) at the optimum pH of 7.5 even at low temperature (below 4 degrees C). This phosphoenzyme is an enzyme-bound, high-energy-phosphate intermediate, because ATP was formed from it on incubation with ADP in the presence of Mg2+ (1 mM). This finding suggests that the phosphoenzyme functions as an intermediate in NDP-kinase action.

Topics: Amino Acids; Animals; Carcinoma, Ehrlich Tumor; Cell Line; Cytosol; HeLa Cells; Humans; Hydrogen-Ion Concentration; Kinetics; Leukemia, Myeloid, Acute; Macromolecular Substances; Mice; Molecular Weight; Nucleoside-Diphosphate Kinase; Phosphorus Radioisotopes; Phosphorylation; Phosphotransferases; Ribonucleotides

Topics: Adenosine Triphosphate; Animals; Carcinoma, Ehrlich Tumor; Cell Nucleus; Cytosol; Enzyme Induction; Indicators and Reagents; Interferons; Mice; Phosphorus Radioisotopes; Phosphorylation; Physarum; Protein Kinases; Spermidine; Spermine

It has been shown previously the 32Pi is incorporated into phosphatidylinositol 30 times faster than into the other phospholipids classes in Ehrlich ascites tumor cells, whereas [1-14C] glycerol is incorporated at almost the same rate (Waku, K., Nakazawa, Y. and MOri, W. (1976) J. Biochem. 79, 407-411). It was therefore suggested that there is a recirculating system (phosphatidylinositol leads to diacylglycerol leads to phosphatidic acid leads to CDP diacylglycerol leads to phosphatidylinositol) of phosphatidylinositol in Ehrlich ascites tumor cells. In this work, 32Pi or [1-3H] glycerol was injected into the peritoneal cavity of mice bearing Ehrlich ascites tumor cells from which the lipids were extracted after selected periods. Phosphatidylinositol was prepared and fractionated in the form of dimethylphosphatidic acid into six molecular species by AgNO3-impregnated TLC. The specific radioactivities of the fractionated species were determined. 32Pi was incorporated into diene molecular species and [1-3H] glycerol into monoene species with a higher rate than the other species and both precursors were incorporated into tetraene species rather slowly. 32P/3H values appeared to be at almost the same for each molecular species, although monoene species showed slightly lower values. These results suggest that there could be a recirculating of the phosphorylinositol moiety in each of the molecular species of phosphatidylinositol.

Topics: Animals; Carcinoma, Ehrlich Tumor; Kinetics; Male; Mice; Phosphatidic Acids; Phosphatidylinositols; Phosphorus Radioisotopes; Tritium

Topics: Amnion; Animals; Carcinoma, Ehrlich Tumor; Cell Line; Female; Humans; Interferons; L Cells; Mice; Phosphorus Radioisotopes; Phosphorylation; Pregnancy; Protein Kinases; Radioisotope Dilution Technique; Ribosomal Proteins; Ribosomes

Topics: Alkyl and Aryl Transferases; Animals; Carbon Radioisotopes; Carcinoma, Ehrlich Tumor; Fatty Acids, Nonesterified; Kinetics; Magnesium; Mice; Microsomes; Phosphorus Radioisotopes; Radioisotope Dilution Technique; Substrate Specificity; Transferases

T4 RNA ligase (EC 6.5.1.3) has been used to link cytidine 3',5'-[5'-32P]bisphosphate or unlabelled cytidine 3',5'-bisphosphate (pCp) covalently to the 3'-OH of individual components of 5'-triphospho-oligo[(2'-5')adenylyl]adenosine [ppp(A2'p)nA with n = 2 or 3] and adenylyl(2'-5')adenylyl(2'-5')adenosine [(A2'p)2A] to yield 5'-triphospho-oligo[(2'-5')adenylyl]adenylyl(3'-5')cytidine 3'-phosphate [ppp(A2'p)nApCp with n = 2 or 3] and adenylyl(2'-5')adenylyl(2'-5')adenylyl(3'-5')cytidine 3'-phosphate (A2'p)2ApCp], respectively. The radioactive products isolated by high-performance liquid chromatography had specific activities greater than 10(6) Ci/mol. These products were found to be effective probes for use in radiobinding and radioimmune assays for ppp(A2'p)nA and (A2'p)nA [M. Knight et al. (1980) Nature 288, 189-192]. ppp(A2'p)nA is unstable in cell-free systems and in intact cells. ppp(A2'p)nApCp, however, was found to be much more stable than ppp(A2'p)nA in extracts of rabbit reticulocytes or Ehrlich ascites tumour cells. (A2'p)2Ap, (A2'p)2ApC (reticulocyte only) or (A2'p)2ApCp were also more stable than unmodified (A2'p)2A in these systems. The results are consistent with a specific degradation pathway for ppp(A2'p)nA which proceeds from the 3' terminus. ppp(A2'p)3ApCp activated the ppp(A2'p)nA-dependent RNase and inhibited protein synthesis in a reticulocyte cell-free system at least as well as unmodified tetramer ppp(A2'p)3A, suggesting that an unmodified 3' terminus is not required for full activity in this system. In extracts from mouse Ehrlich ascites tumour or L-cells, however, ppp(A2'p)nApCp was greater than or equal to 30 fold less active (if directly active at all).

Topics: Adenine Nucleotides; Animals; Carcinoma, Ehrlich Tumor; Cell-Free System; Cytosine Nucleotides; Drug Stability; In Vitro Techniques; Isotope Labeling; Oligonucleotides; Oligoribonucleotides; Phosphorus Radioisotopes; Protein Biosynthesis; Rabbits; Reticulocytes

Topics: Animals; Carcinoma, Ehrlich Tumor; Chromatography, Agarose; Chromatography, Gel; Chromatography, Ion Exchange; Liver; Mice; Phosphorus Radioisotopes; RNA

In view of results of extracorporal measurements of the 32 P-incorporation into irradiated or nonirradiated tumors biochemical investigations of the 32P-incorporation have been performed, aiming at comprehension of the meaning and at clarification of the interpretability of these measurements. The results show that irradiation mostly decreases the content of 32P in nucleic acids. In principle, however, the latter is so low as compared to the total content of 32P, especially three hours after 32P-injection, that variations are hardly to be recorded by means of extracorporal measurements, particularly because they are superposed by changes in the 32P-content of other fractions.

Topics: Animals; Carcinoma, Ehrlich Tumor; Female; Male; Mice; Nucleic Acids; Phosphates; Phosphorus Radioisotopes; Time Factors

32P-Labeled phospholipids with specific activities up to 400 mCi/mmole as well as [32P]CDP-choline were prepared by cultivation of mouse fibroblasts or mouse Ehrlich ascites cells in the presence of [32P]orthophosphate. The method was also used to prepare [methyl-3H]choline-labeled glycerophospholipids from [3H]choline. The yields and the specific activities of the phospholipids were significantly lower when preparations of ox white blood cells were used.

Topics: Animals; Carcinoma, Ehrlich Tumor; Cattle; Cell Line; Cell Transformation, Neoplastic; Chromatography, Thin Layer; Cytidine Diphosphate Choline; Erythrocytes; Fibroblasts; Humans; Isotope Labeling; Leukocytes; Mice; Phosphatidic Acids; Phosphatidylcholines; Phospholipids; Phosphorus Radioisotopes; Structure-Activity Relationship; Tritium

Topics: Animals; Carcinoma, Ehrlich Tumor; Chickens; Cobalt Radioisotopes; Dose-Response Relationship, Radiation; Electrophoresis; Erythrocytes; Gamma Rays; Humans; Male; Mice; Phosphorus Radioisotopes; Radiation Effects; X-Rays

Lactic dehydrogenase virus (LDV) was purified from infectious ascites fluid of mice bearing Ehrlich tumours using Sepharose gel filtration and rate zonal and isopycnic sedimentation. In glycerol gradients, a sedimentation coefficient of about 200S and a buoyant density of 1-14 g/ml was determined for the virus particle. Spherical particles with diam. between 62 and 80 nm, depending on the method of fixation and staining, have been identified electron microscopically. The virus particle consists of a spherical nucleocapsid wrapped into a double-layered envelope. The nucleocapsids, isolated by treatment with NP40 and purified by centrifuging on sucrose gradients had a sedimentation coefficient of 176S. Electron micrographs show spherical particles with a diam of 35 plus or minus nm. Classification of LDV as a member of the togaviridae family is discussed.

Topics: Animals; Ascitic Fluid; Carcinoma, Ehrlich Tumor; Centrifugation, Density Gradient; Chromatography, Gel; Lactate dehydrogenase-elevating virus; Mice; Microscopy, Electron; Nucleoproteins; Phosphorus Radioisotopes; Phosphotungstic Acid; RNA Viruses; Sindbis Virus; Staining and Labeling; Viral Proteins

N-Bromosuccinimide cleavage of in vivo 32P-labelled lysine-rich histone isolated from rapidly dividing cells has been studied. N-Bromosuccinimide cleaves F1-histone into two fragments, a small N-terminal piece and a larger C-terminal portion. The phosphate-induced microheterogeneity and associated radioactivity which has been linked to cell replication, is found in the carboxyterminal fragment, No phosphorous is found associated with the amino-terminal fragment when histone phosphorylation is associated with cell division. The specific tryptic phosphopeptides obtained from in vivo labelled F1 are clearly different from those obtained from in vitro incubations of free F1-histones and cytoplasmic protein kinase.

Topics: Animals; Carcinoma, Ehrlich Tumor; Cell Division; Female; Histones; Lysine; Mice; Peptide Fragments; Phosphates; Phosphopeptides; Phosphorus Radioisotopes; Protein Kinases; Succinimides; Trypsin

Topics: Animals; Carcinoma, Ehrlich Tumor; Cell Division; Deoxyuridine; DNA, Neoplasm; Phosphates; Phosphorus Radioisotopes; Thymidine; Vinyl Compounds

Topics: Acetone; Alcohol Oxidoreductases; Animals; Binding, Competitive; Borohydrides; Carcinoma, Ehrlich Tumor; Centrifugation, Density Gradient; Chromatography, Gel; Chromatography, Ion Exchange; Deoxycholic Acid; Drug Stability; Electrophoresis, Paper; Guinea Pigs; Kinetics; Membranes; Mice; Microsomes; Mitochondria, Liver; NAD; NADP; Niacinamide; Organophosphorus Compounds; Palmitic Acids; Phosphatidic Acids; Phosphorus Radioisotopes; Structure-Activity Relationship; Time Factors; Tritium

Topics: Adenosine Diphosphate; Adenosine Triphosphatases; Adenosine Triphosphate; Animals; Calcium; Carcinoma, Ehrlich Tumor; Cell Fractionation; Cell Line; Mitochondria; Oxidative Phosphorylation; Oxygen Consumption; Phosphorus Radioisotopes

Topics: Adenosine Triphosphatases; Animals; Ascitic Fluid; Carcinoma, Ehrlich Tumor; Carcinoma, Hepatocellular; Cell Fractionation; Dinitrophenols; Female; Hepatectomy; Leukemia L1210; Liver; Liver Neoplasms; Liver Regeneration; Male; Mice; Mitochondria, Liver; Neoplasms, Experimental; Oxidative Phosphorylation; Oxygen Consumption; Phosphates; Phosphorus Radioisotopes; Rats; Uncoupling Agents

Topics: Adenine Nucleotides; Animals; Benzoates; Carcinoma, Ehrlich Tumor; Cell Nucleus; Cellulose; Centrifugation, Zonal; Chromatography; Cytoplasm; DNA; Formaldehyde; Mengovirus; Nucleic Acid Denaturation; Nucleotides; Phosphorus Radioisotopes; Poly A-U; Poly U; Polynucleotides; RNA, Neoplasm; RNA, Ribosomal; RNA, Viral

Topics: Animals; Carbon Radioisotopes; Carcinoma, Ehrlich Tumor; Cell Fractionation; Centrifugation, Density Gradient; Culture Techniques; Cycloheximide; Cytoplasm; Dactinomycin; DNA-Directed RNA Polymerases; Mice; Nucleoproteins; Parainfluenza Virus 1, Human; Phosphorus Radioisotopes; Ribonucleases; RNA, Viral; Temperature; Transcription, Genetic; Viral Proteins; Virus Replication

Topics: Adenosine Triphosphate; Animals; Carcinoma, Ehrlich Tumor; Cell Membrane; Cells, Cultured; Culture Media; Cyclic AMP; Histones; Mice; Microscopy, Phase-Contrast; Phosphates; Phosphorus Radioisotopes; Protein Kinases; Proteins

Topics: Animals; Carbon Radioisotopes; Carcinoma, Ehrlich Tumor; Centrifugation, Density Gradient; Culture Techniques; Cytoplasm; Electrophoresis, Polyacrylamide Gel; Fibroblasts; Mice; Mice, Inbred BALB C; Microscopy, Electron; Molecular Weight; Nucleoproteins; Parainfluenza Virus 1, Human; Peptides; Phosphorus Radioisotopes; Ribonucleases; RNA, Viral; Tritium; Viral Proteins

Topics: Adenosine Triphosphate; Amino Acids; Animals; Carcinoma, Ehrlich Tumor; Cell Membrane; Cells, Cultured; Chromatography, Ion Exchange; Cyclic AMP; Mice; Organophosphorus Compounds; Phosphoproteins; Phosphorus Radioisotopes; Serine; Threonine

Topics: Animals; Base Sequence; Carcinoma, Ehrlich Tumor; Cell Nucleus; Chromatography, Gel; Chromatography, Ion Exchange; Electrophoresis, Polyacrylamide Gel; Hot Temperature; Hydroxyapatites; Mice; Nucleic Acid Denaturation; Nucleic Acid Hybridization; Nucleic Acid Renaturation; Pancreas; Phosphorus Radioisotopes; Ribonucleases; Ribonucleotides; RNA, Messenger; RNA, Neoplasm; Temperature; Transcription, Genetic

Topics: Animals; Base Sequence; Carcinoma, Ehrlich Tumor; Cell Nucleus; Chromatography, Gel; Chromatography, Ion Exchange; Drug Stability; Hot Temperature; Hydroxyapatites; Mice; Nucleic Acid Denaturation; Nucleic Acid Hybridization; Oligonucleotides; Osmolar Concentration; Phosphorus Radioisotopes; Polyribosomes; Ribonucleases; Ribonucleotides; RNA, Messenger; RNA, Ribosomal

Topics: Animals; Carcinoma, Ehrlich Tumor; Cell Differentiation; Cell Division; Cell Transformation, Neoplastic; Cells; DNA, Neoplasm; Kinetics; Mice; Microwaves; Mitosis; Phosphorus; Phosphorus Radioisotopes; Radiation Effects; Temperature

Topics: Animals; Carcinoma, Ehrlich Tumor; Cell Nucleus; Chromatography, Gel; Chromatography, Ion Exchange; Hydrolysis; Hydroxyapatites; Isotope Labeling; Mice; Oligonucleotides; Phosphorus Radioisotopes; Ribonucleotides; RNA, Messenger; RNA, Neoplasm; Spectrophotometry, Ultraviolet; Time Factors

Topics: Animals; Carcinoma, Ehrlich Tumor; Cell Line; Cell Nucleus; Cricetinae; Cytarabine; Dactinomycin; Deoxyadenosines; Electrophoresis, Polyacrylamide Gel; Half-Life; Kidney; Magnesium; Methionine; Mice; Molecular Weight; Phosphates; Phosphorus Radioisotopes; RNA; RNA, Ribosomal; Spectrophotometry, Ultraviolet; Temperature; Time Factors; Tritium; Uridine

Topics: Animals; Arginine; Aspartic Acid; Carbon Radioisotopes; Carcinoma, Ehrlich Tumor; Cell Nucleus; DNA; Electrophoresis, Starch Gel; Glutamates; Histones; Hydroxyurea; Lysine; Mice; Mice, Inbred BALB C; Microscopy, Phase-Contrast; Phosphorus Radioisotopes; Time Factors

Topics: Alkaline Phosphatase; Animals; Carbon Radioisotopes; Carcinoma, Ehrlich Tumor; Cattle; Centrifugation, Density Gradient; Chromatography, Gel; Chromatography, Ion Exchange; Chromatography, Thin Layer; DNA, Neoplasm; Hot Temperature; Intestines; Isotope Labeling; Kinetics; Mice; Nucleic Acid Denaturation; Nucleic Acid Hybridization; Pancreas; Phosphorus Radioisotopes; Ribonucleases; RNA, Neoplasm; Thymidine; Time Factors; Tritium; Uridine

Topics: Animals; Carcinoma, Ehrlich Tumor; Cell Nucleus; Cells, Cultured; Cytarabine; Cytosine Nucleotides; Depression, Chemical; DNA Nucleotidyltransferases; DNA, Neoplasm; Kinetics; Mice; Phosphorus Radioisotopes; Thymine Nucleotides; Time Factors

Topics: Animals; Calcium; Calcium Radioisotopes; Carcinoma, Ehrlich Tumor; Cell Fractionation; Cells; Lipoproteins; Mice; Phosphorus; Phosphorus Radioisotopes; Protein Binding

Topics: Adenosine Diphosphate; Adenosine Monophosphate; Adenosine Triphosphate; Adenylyl Cyclase Inhibitors; Adenylyl Cyclases; Animals; Binding Sites; Carcinoma, Ehrlich Tumor; Chromatography, Thin Layer; Epinephrine; Female; Fluorides; Kinetics; Organometallic Compounds; Phosphorus Radioisotopes; Pregnancy; Protein Binding; Structure-Activity Relationship; Sulfur; Tritium

Topics: Adenosine Triphosphatases; Animals; Binding Sites; Carcinoma, Ehrlich Tumor; Cell Fractionation; Cell Membrane; Centrifugation, Density Gradient; Chelating Agents; Kinetics; Magnesium; Mice; Microscopy, Electron; Ouabain; Phosphorus Radioisotopes; Potassium; Protein Binding; Receptors, Drug; Sodium; Spectrophotometry; Zinc

Topics: Animals; Carbon Radioisotopes; Carcinoma, Ehrlich Tumor; Cell Nucleolus; Cells, Cultured; Centrifugation, Density Gradient; Chick Embryo; Cycloheximide; Cytoplasm; Dactinomycin; Lung; Mice; Microscopy, Electron; Nucleoproteins; Parainfluenza Virus 1, Human; Phosphorus Radioisotopes; RNA, Viral; Time Factors; Tritium; Uridine; Viral Proteins

Topics: Animals; Carcinoma, Ehrlich Tumor; Cells, Cultured; Mice; Neoplasm Proteins; Oxidative Phosphorylation; Phosphorus Radioisotopes; Ribosomes; Time Factors

Topics: Animals; Carcinoma, Ehrlich Tumor; Female; Methods; Mice; Phosphorus Radioisotopes; Radiotherapy Dosage