scholarly journals Use of actinomycin D for the specific quenching of fluorescence of deoxyribonucleic acid in cells stained with acridine aminoderivatives.

1976 ◽  
Vol 24 (11) ◽  
pp. 1169-1172 ◽  
Author(s):  
A V Zelenin ◽  
E A Kirianova ◽  
V A Kolesnikov ◽  
N G Stepanova
Keyword(s):  
Acridine Orange ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Actinomycin D ◽  
Quenching Of Fluorescence ◽  
Approximate Estimation ◽  
In Cells

Actinomycin D specifically quenches the fluorescence of acridine orange and quinacrine bound to deoxyribonucleic acid in cytologic preparations, but does not change the fluorescence of these fluorochromes bound to RNA. The following fluorescence-cytochemical applications of techniques based on these findings can be suggested: (a) distinction between deoxyribonucleic acid and ribonucleic acid; (b) detection of double-stranded virus ribonucleic acid; (c) approximate estimation of the lengths of A-T sequences in deoxyribonucleic acid molecules.

scholarly journals Indirect fluorescence of primary and secondary myofibers in developing porcine muscle.

1977 ◽  
Vol 25 (6) ◽  
pp. 439-442 ◽  
Author(s):  
D H Beermann ◽  
R G Cassens
Keyword(s):  
Skeletal Muscle ◽  
Acridine Orange ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Porcine Muscle ◽  
Structural Framework ◽  
Linear Sequence ◽  
Cryostat Sections ◽  
Two Populations ◽  
Fetal Muscle

Cytochemical differentiation of two populations of developing skeletal myofibers has been demonstrated in fetal muscle with metachromatic fluorescence of ribonucleic acid and deoxyribonucleic acid by staining fresh frozed cryostat sections of developing porcine skeletal muscle with acridine orange (CL. 46005). Evidence is presented that supports the hypothesis that first-formed myofibers (primary myofibers) serve as a structural framework upon which myoblasts proliferate, fuse in linear sequence and give rise to a second population (secondary myofibers) of myofibers.

scholarly journals Deoxyribonucleic acid-dependent ribonucleic acid polymerases from normal and polyoma-transformed BHK-21/C13 cells

1975 ◽  
Vol 145 (3) ◽  
pp. 509-516 ◽  
Author(s):  
R J Cooper ◽  
H M Keir
Keyword(s):  
Ionic Strength ◽  
Rna Polymerase ◽  
Dna Content ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Actinomycin D ◽  
Activity Ratio ◽  
Cell Types ◽  
Dna Templates ◽  
Alpha Amanitin

DNA-dependent RNA polymerase (EC 2.7.7.6) ACTIVITIES FROM NORMAL BHK-21/C13 cells and from BHK-21/C13 cells transformed by polyoma virus (PYY cells) were solubilized and fractionated on columns of DEAE-Sephadex. Various properties of the A and B enzymes from the two types of cell were compared. 1. The yields of polymerase relative to the DNA content of the nuclear preparations are similar for both cell types. 2. The ionic-strength optima of polymerases A and B are 12.5 mM and 100mM with respect to (NH4)2SO4 for both cell types. 3. The Mn2+/Mg2+ activity ratio (measured at the respective optimum for each cation) for polymerase A from BHK-21/C13 cells was 1.48 and for the polymerase A from PYY cells was 0.55. The corresponding ratios for polymerase B were 10.11 for BHK-21/C13 cells and 22.75 for PYY cells. 4. Minor differences in the ability of the A polymerases to transcribe native and denatured DNA templates were observed; such differences were not apparent when the B polymerases were compared. 5. All the polymerases were inhibited completely by actinomycin D and by rifampicin AF/013, but not markedly so by rifampicin. Alpha-amanitin inhibited polymerase B but not polymerase A.

scholarly journals Nuclear-Cytoplasmic Relationships in Human Cells in Tissue Culture

1959 ◽  
Vol 6 (1) ◽  
pp. 1-6 ◽  
Author(s):  
Lester Goldstein ◽  
Julie Micou
Keyword(s):  
Tissue Culture ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Human Cells ◽  
Human Amnion ◽  
Progressive Movement ◽  
Excess Amount ◽  
In Cells

The movement of ribonucleic acid (RNA) from nucleus to cytoplasm has been studied, by autoradiographic techniques, in cells of the human amnion grown in tissue culture. Cells were exposed to cytidine-H3 for 1 hour after which time only the RNA of the nuclei was labelled. After this 1 hour exposure the cells were placed in a medium containing an excess amount of unlabelled cytidine. Periodically, cells from this medium were fixed. Autoradiographs showed that there was a progressive movement of the label from nucleus to cytoplasm, such that after 24 hours essentially all the label was in the RNA of the cytoplasm. A study of the incorporation of the cytidine-H3 in deoxyribonucleic acid (DNA), in the same population of cells at the same times, indicated that the presence of excess amounts of unlabelled cytidine almost instantaneously inhibited further utilization of cytidine-H3. It is concluded that RNA moves from nucleus to cytoplasm as a complex polynucleotide structure.

Permeability increases in cells of Salmonella enteritidis induced by ethylenediaminetetraacetate and temperature treatments

1972 ◽  
Vol 18 (12) ◽  
pp. 1803-1807 ◽  
Author(s):  
J. R. Chipley ◽  
H. M. Edwards Jr
Keyword(s):  
Ribonucleic Acid ◽  
Cell Walls ◽  
Untreated Control ◽  
Salmonella Enteritidis ◽  
Deoxyribonucleic Acid ◽  
Actinomycin D ◽  
Temperature Dependent ◽  
Lethal Effects ◽  
Treated Cell ◽  
Uptake And Release

Treatment of cells of Salmonella enteritidis with ethylenediaminetetraacetate (EDTA) resulted in losses of less than 4% of the total cellular deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and protein. No lethal effects could be observed when treated cells were plated and counted. The uptake of 14C-actinomycin-D, 22Na+, and 65Zn2+ into EDTA-treated cells and the release of 65Zn2+ from EDTA-treated cell walls were strictly temperature-dependent. The uptake of 22Na+ and 65Zn2+ in control and EDTA-treated cells appeared to be enzymatically controlled and could be inhibited by p-chloromercuribenzoate. The uptake and release of these radioisotopes in treated cells was two to five times that of untreated, control cells. The release of lipopolysaccharide could be correlated with a change in permeability of cells when they were treated with EDTA.

Effect of Some Metabolic Inhibitors on Vinblastine-Induced Ribosomal-Granular Material Complexes

1971 ◽  
Vol 29 ◽  
pp. 548-549
Author(s):  
Awtar Krishan ◽  
Dora Hsu
Keyword(s):  
Granular Material ◽  
Rna Synthesis ◽  
Culture Medium ◽  
Actinomycin D ◽  
Metabolic Inhibitors ◽  
Protein And Rna Synthesis ◽  
Apparent Reduction ◽  
Plant Alkaloids ◽  
In Cells

Cells exposed to antitumor plant alkaloids, vinblastine and vincristine sulfate have large proteinacious crystals and complexes of ribosomes, helical polyribosomes and electron-dense granular material (ribosomal complexes) in their cytoplasm, Binding of H3-colchicine by the in vivo crystals shows that they contain microtubular proteins. Association of ribosomal complexes with the crystals suggests that these structures may be interrelated.In the present study cultured human leukemic lymphoblasts (CCRF-CEM), were incubated with protein and RNA-synthesis inhibitors, p. fluorophenylalanine, puromycin, cycloheximide or actinomycin-D before the addition of crystal-inducing doses of vinblastine to the culture medium. None of these compounds could completely prevent the formation of the ribosomal complexes or the crystals. However, in cells pre-incubated with puromycin, cycloheximide, or actinomycin-D, a reduction in the number and size of the ribosomal complexes was seen. Large helical polyribosomes were absent in the ribosomal complexes of cells treated with puromycin, while in cells exposed to cycloheximide, there was an apparent reduction in the number of ribosomes associated with the ribosomal complexes (Fig. 2).

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