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Biomedical subjects

R Braun

Publications and source records attributed to R Braun.

At least 199 records · Page 11Linked to original sources
PubMed1978

Ribosomal DNA in spores of Physarum polycephalum.

DNA was isolated from plasmodia, spores and newly hatched amoebae of the slime mould Physarum polycephalum. The DNA preparations were fractionated in CsCl gradients and each fraction hybridised to combined 19 S + 26 S rRNA. In all three DNA preparations hybridisation was found to be limited to satellite DNA (rho = 1.714 g/cm3) and at saturation was found to reach a level of 0.16--0.18 % of total DNA. The main band of nuclear DNA (rho = 1.702 g/cm3) did not hybridise appreciably. Further experiments using analytical CsCl gradients revealed that the ratio of satellite to main band DNA was similar in all three preparations. It is concluded that the genes for ribosomal RNA are equally reiterated in spores, hatching amoebae and in plasmodia. They appear to be similarly organised in all stages of the life cycle examined so far.

Alleles↗
PubMed1978

The influence of a new antitumor agent on hemopoietic colony forming cells.

The cytostatic and immunsuppressive agent N'-methyl-N'-beta-chloroethylbenzaldehyde hydrazone (B1) in in-vitro experiments has a stimulating effect on colony-forming culture (CFUc) of bone marrow from C57BL mice. This unusual behaviour, which is in contrast to other cytostatics, could also be observed in vitro with CFUc obtained from mice treated with therapeutic doses of B1 for 2 weeks. This stimulation is not a particular effect of B1 alone but seems to depend on a synergistic effect of the combination of B1 and the colony-stimulating activity (CSA) present in the serum from endotoxin-treated mice (MP) in the testing system. The results suggest that the described effect of B1 is due to an interference at the cell membrane of CFUc or their precursor cells.

Antineoplastic Agents↗
PubMed1978

A new method for the purification of RNA polymerase II (or B) from the lower eukaryote Physarum polycephalum. The presence of subforms.

The DNA-dependent RNA polymerase II or B from the lower eukaryote Physarum polycephalum has been purified to apparent homogeneity by a new method employing poly(ethylene imine) precipitation and elution, and heparin-Sepharose affinity chromatography. The method is readily scaled up or down and affords a purification of over 5000-fold with a yield of 35-45%. The procedure is easy to perform and can be carried out in less than three days even on a large scale. Furthermore, it gives enzyme of higher purity and in at least 10-fold greater yield than previously published procedures for its purification from this organism. These improvements have allowed the detection of a series of subforms of the enzyme. The combination of precipitation using poly(ethylene imine) with chromatography on heparin-Sepharose may prove useful in the preparation of other proteins which interact with nucleic acids.

Chromatography, Affinity↗
PubMed1978

Establishment of a variant cell clone growing at 41 degrees C from embryonic rat and tupaia (tree shrew) fibroblast cells.

Rat and tupaia 41 degrees C temperature variant cell clones were derived from parental embryonic cells, cloned and established in tissue cultures. Both variant cell clones grew permanently at 41 degrees C. The morphology of these cell clones was altered in comparison to the original fibroblast cell clones. The cell biological characterization of the rat and tupaia 41 degrees C temperature variant cell clones showed that both cell clones were stable. After abolishing the selection pressure (incubation at 41 degrees C) for more than 10 further cell passages by incubation at 37 degrees C and then raising the temperature again to 41 degrees C, neither of the cell clones lost their newly acquired property of growing at 41 degrees C. This fact demonstrates that the newly acquired property is certain to be genetically manifest in both cell clones. The modal number of chromosomes of the rat 41 degrees C temperature variant cell clone was increased, and in the case of the tupaia variant cell clone, bimodality was observed. The plating efficiency of both cell clones did not rise significantly in comparison to the parental cells. Neither of the 41 degrees C temperature variant cell clones grew in semi-solid medium.

Animals↗
PubMed1978

Metabolic stability of the extrachromosomal ribosomal RNA genes in the slime mould Physarum polycephalum.

The rRNA genes of the slime mould Physarum polycephalum are located on free, linear DNA molecules of a discrete size, Mr=38X10(6). Using an isotope dilution technique we have examined the metabolic stability of these extrachromosomal genes during active, balanced growth. Microplasmodia, prelabelled with [3H]thymidine, were used to prepare synchronous surface plasmodial cultures which were subsequently grown on unlabelled medium. The gross synthesis of ribosomal DNA was then determined over three consecutive mitotic divisions from the ratio of 3H to 14C in a hybrid formed between the extracted ribosomal [3H]DNA and a [14C]rRNA probe. It was found that ribosomal DNA, like chromosomal DNA, is completely stable during active growth.

Drug Stability↗
PubMed1978

Degradation of the ribosomal genes by DNAse I in Physarum polycephalum.

Treatment of nuclei from Physarum polycephalum with DNAse I leads to DNA fragments with a regular pattern of multiples of 10 nucleotides, when analyzed on gels under denaturing conditions as has been shown for other eukaryotes. Reports from Weintraub and Axel lead to the conclusion, that active genes are preferentially digested by DNAse I. When Physarum chromatin is degraded by DNAse I, the ribosomal genes are no longer available for hybridization with 19-S and 26-S rRNA and are thus preferentially destroyed. Degradation of chromatin from nuclei in mitosis, where no rRNA is synthesized and from nuclei in late G2 phase, where rRNA synthesis is maximal, leads to the same proportion of the ribosomal sequences being lost for hybridization. Therefore the preferential degradation of the ribosomal genes in Physarum by DNAse I probably does not reflect the actual momentary activity of these genes. This suggests that DNAse I treatment may distinguish between active chromatin and very strongly repressed chromatin.

Cell Nucleus↗
PubMed1978

[Effect of N'-methyl-N'-beta-chlorethylbenzaldehyde hydrazone on aerobic glycolysis in Ehrlich ascites cells].

N'-Methyl-N'-beta-chloroethylhydrazones are potent inducers of aerobic glycolysis of Ehrlich ascites carcinoma (E.A.) cells without influencing directly the respiration which is reduced only after glycolysis is at its maximum. This induction of glycolysis is not influenced by simultaneous blocking of respiration by potassium cyanide. A primary attack on the cell membrane leading to an increased glucose influx is discussed because of the in vitro inhibition of these beta-chloroethylhydrazones of glyceraldehyde-3-phosphate oxidoreductase.

Animals↗
PubMed1978

Inhibition of nucleoside uptake into Ehrlich ascites carcinoma cells by new cytostatic methylhydrazones.

For the purpose of studying the structure-activity relationships of N'-methyl-N'-beta-chloroethylbenzaldehyde hydrazones, new hydrazones were synthesized in which the beta-chloroethyl group was replaced by substituents conveying a partial positive charge at the N' moiety by induction and/or mesomerism. In a preliminary antitumor evaluation, some of these hydrazones showed a cytostatic activity in vivo similar to that of the beta-chloroethyl hydrazones. The new hydrazones also strongly inhibit the uptake of nucleosides into tumor cells and intensify the in vivo cytostatic effect of methotrexate.

Animals↗
PubMed1977

The replication of ribosomal DNA in Physarum polycephalum.

The DNA coding for ribosomal RNA in Physarum polycephalum exists as a collection of extra-chromosomal molecules of molecular weight 37 X 10(6). We have investigated the replication of rDNA, with the following results. (a) Replication of rDNA is unscheduled. This means that molecules that are replicated at any particular time in one mitotic cycle have an equal probability of replicating again in each time interval in the subsequent cycle. Similarly, in a single cycle, some molecules replicate more than once, and some not at all. (b) Replication forks appear to move bidirectionally from points 45% or 33% from one end of the DNA. Replicating molecules observed by electron microscopy are all linear.

DNA Replication↗
PubMed1977

Synthesis and transport of myosin in Physarum polycephalum.

Immunological techniques have been used to study the rate of synthesis and intracellular transport of myosin in the slime mould Physarum polycephalum. Quantitative precipitation of myosin in homogenates of Physarum was achieved using an antimyosin antibody produced in rabbit in response to purified Physarum myosin. Dodecylsulphate-gel electrophoresis revealed that about 50% of the precipitated material is myosin. The rates of synthesis of total cellular protein and myosin were measured over the mitotic cycle. Both were found to increase exponentially or linearly between two successive nuclear divisions. Similarly, no difference in the proportion of myosin-synthesising polysomes, assayed by precipitation with antimyosin serum, could be detected between the S phase and G2 phase of the mitotic cycle. Myosin makes up nearly 2% of total plasmodial proteins. Its transport into the nucleus occurs predominantly during the G2 phase.

Animals↗
PubMed1977

The organisation of genes for transfer RNA and ribosomal RNA in amoebae and plasmodia of Physarum polycephalum.

1. Using hybridisation techniques nuclei from both amoebae and plasmodia of Physarum polycephalum were found to contain 275 genes each coding for 5.8-S, 19-S and 26-S rRNA, 685 genes for 5-S rRNA and 1050 genes for tRNA. 2. Hybridisation of these RNA species to both amoebal and plasmodial DNA fractionated on CsCl gradients reveal that the 5.8-S, 19-S and 26-S rRNA genes are located at a satellite position (formula: see text) with respect to the main band of DNA, whereas 4-S RNA genes are located exclusively in the main band of DNA (formula: see text). 3. This result was confirmed by demonstrating that only the 5.8-S, 19-S and 26-S rRNA species hybridise to purified plasmodial ribosomal DNA. 4. The 19-S and 26-S rRNA genes of amoebae are located on extrachromosomal DNA molecules of a discrete size (Mr = 38 X 10(6)) with identical properties to plasmodial ribosomal DNA.

Cell Nucleus↗