Effect of selenium on the growth of three human colon cancer cell lines

The effects of selenium were investigated on three human colon cancer cell lines: Caco 2, HRT 18, and HT 29. At low concentrations (10-100 nM), selenium stimulated cell growth in serum-free medium. Thus, selenium is an essential trace element for cell proliferation. At higher concentrations, selenium inhibited cell growth. The rate of 75Se uptake was the same in all of the cell lines studied, but the quantity incorporated differed. GSH-Px activity was dependent on the selenium content of the medium. DNA and protein synthesis paralleled the growth curve. Comparison with the curve of viability revealed that selenium inhibited cell growth in two ways: by inhibiting DNA synthesis, without affecting cell viability, and, at higher doses, by cytotoxicity.     

Direct selection for curing and deletion of Rhizobium plasmids using transposons carrying the Bacillus subtilis sacB gene

We have constructed derivatives of the transposon Tn5 carrying the mob site (oriT) of plasmid RP4, and an nptI-sacB-sacR cassette [Ried and Collmer, Gene 57 (1987) 239-246]. The mob site, in conjunction with the antibiotic-resistance markers carried on the transposons, allows identification of transposon inserts in cryptic plasmids by mobilisation to other strains. The sacB-sacR genes allow direct selection for the loss or curing of plasmids, because only strains which no longer contain an active sacB gene are able to grow on media containing sucrose. We have tested these transposons in four strains of Rhizobium leguminosarum and two strains of Rhizobium meliloti, and have been able to demonstrate curing of several large cryptic plasmids, and generation of large deletions in many other plasmids. This method has enabled us to show that the R. leguminosarum plasmids pRL12JI and pR1eVF39f carry auxotrophic markers, and that the plasmid pR1eVF39c carries genes which affect colony morphology.     

Protection of the monoamine oxidase in brain synaptosomes by fat- and water-soluble antioxidants during lipid peroxidation

The protective effects of alpha-tocopherol, carnosine and their mixtures on monoamine oxidase activity, accumulation of lipid peroxidation products, lipid fatty acid composition, hydrophobicity and microviscosity of synaptic membranes during lipid peroxidation were studied. It was shown that the protective efficiency is more higher when the mixture of water and liposoluble antioxidants was used.     

Effects of adenosine deaminase on the sensitivity of glucose transport, glycolysis and glycogen synthesis to insulin in muscles of the rat

1. Soleus, extensor digitorum longus (EDL) or hemi-diaphragm muscles of the rat were incubated in the presence of insulin and rates of the processes of glycolysis and glycogen synthesis were measured. 2. The concentrations of insulin required to cause half-maximal stimulation of glycolysis in both soleus and EDL preparations were significantly decreased by the presence of adenosine deaminase in the medium. 3. Adenosine deaminase increased the sensitivity of the process of hexose transport to insulin (in an identical manner to the change in sensitivity of glycolysis) in the EDL preparation. 4. None of the adenosine mediated effects on insulin-stimulated rates of glycolysis were observed in the hemi-diaphragm preparation or on the rates of glycogen synthesis in any of the three muscle preparations. 5. Therefore, changes in the adenosine system in skeletal muscle influence insulin sensitivity regardless of fibre type composition of the muscle.     

Linkage analysis of the murine mos proto-oncogene on chromosome 4

A linkage analysis of the murine Mos gene, which codes for the c-mos proto-oncogene, was performed in 88 backcross progeny of an interspecies cross of laboratory mice and Mus spretus. Linkage was tested for four different genes on mouse chromosome 4: Aco-1, Mup-1, b, and Ifb. The gene order (from centromere) with intervening percentage recombination is Mos-15.9 (+/- 3.9)-Aco-1-5.6 (+/- 2.4)-Mup-1-3.4 (+/- 1.9)-b-5.6 (+/- 2.4)-Ifb. These results confirm the previous assignment of Mos to chromosome 4 on the basis of segregation in somatic cell hybrids (D. Swan et al., 1982, J. Virol. 44: 752-754) and show furthermore that Mos and the Ifa/Ifb clusters are not tightly linked as a group of intronless genes, but are separated by a map distance of 30.6 +/- 4.9 recombination units. The linkage data obtained in the present study place Mos in a region compatible with the physical map (D. W. Threadgill and J. E. Womack, 1988, Genomics 3: 82-86).     

Action of heparin on protein fractions of isolated nuclei and on their DNA content.

Isolated nuclei of rat hepatocytes were incubated with 0.05% sodium heparinate for 2 to 10 min. Alterations in the nuclei were controlled biochemically, determining the amounts of DNA and histones, and by cytophotometric methods determining the amounts of total and nonhistone proteins and DNA. Under the selected experimental conditions 95% of histones are bound already after 5-min incubation with heparin; nonhistone proteins of cell nuclei remain unchanged. The blockade of histones is followed by DNA diffusion into the incubation medium. Experiments with nuclear staining with alcian blue proved the specificity of heparin binding with histones and showed that heparin-histone complex remains in the nuclei, and its histones lose their extractability with 0.25 n HCl.