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Biomedical subjects

F R Leach

Publications and source records attributed to F R Leach.

At least 37 records · Page 2Linked to original sources

Growth of animal cells on glass fiber filters and their utilization for biosynthetic analysis.

HeLa and L-M cells can be effectively grown directly on glass fiber filters to yield replicate cultures that allow easy analysis of biosynthetic capabilities through measurement of radioactive precursor uptake and incorporation. The glass fiber filters are superior to glass cover slips, growth in scintillation vials, and growth on Millipore filters in the ease of handling during experimental treatment and in the amount of radioactivity incorporated during the labeling period. These parameters are experimentally established and a typical application of the procedure that demonstrates the hydroxyurea inhibition of DNA synthesis is presented.

Cell Count↗

Effects of addicting and nonaddicting drugs on HeLa cells.

After 70 subcultivations of HeLa cells in the presence of amphetamine, diphenhydramine hydrochloride, caffeine, codeine, or morphine, cells were obtained with increased tolerance to the respective drug. Withdrawal of amphetamine, diphenyhydramine hydrochloride or morphine from cells cultivated 645 days in the presence of the drug yielded cells that had either a decreased growth rate in the absence of drug or superior plating efficiencies with low drug concentration when compared to untreated HeLa cells. These observations would be expected from addicted cells. Cinematographic observations showed cell membrane undulation decreased in the presence of morphine. The membrane undulations of cells tolerant to morphine were less sensitive to that drug.

Cell Count↗

The effect of Waring Blendor treatment on transformation in Bacillus subtilis.

Treatment of competent Bacillus subtilis in a Waring Blendor for 10 s increases transformability of the culture about twofold while reducing the attachment of DNA to competent cells by 80%. The effectiveness of attached DNA in producing transformants is increased 10-fold by this treatment. The uptake of transforming DNA into a DNase-resistant state is progressively reduced by 70% during a 120-s blending treatment. Blending for 30-45 s diminishes transformability to about 10% of the original nonblended value without affecting the viable cell titer. No effect is produced by 30 s of blending on transformability if the irreversible uptake of DNA has been completed. Thus, the inhibition occurs at an early step in the transformation sequence. Treatment of the competent culture for 60 s or longer in the Waring Blendor reduces both the number of transformants obtained and the total number of viable cells.

Bacillus subtilis↗

15-Azasteroid blockage of cell permeability and mitochondrial respiration.

The 15-azasteroid, 1,10,11,11a-tetrahydro-11a-methyl-2H naphth (1,2-g)indol-7-o1, inhibits the growth of the cell culture lines KB and L-M as well as several strains of bacteria. The inhibition of growth is reversed following removal of the steroid from the growth medium. Using in vitro grown L-M cells, the compound inhibited the transport of amino acids and uracil. The action was non-detergent like and at least 100 times more effective in terminating metabolite transport than sodium azide. The azasteroid inhibited the oxidation of glutamate in isolated rat liver mitochondria. The oxidation of succinate was not effected by the azasteroid alone but in the presence of glutamate, the azasteroid uncoupled the oxidation of succinate from the ADP-ATP control. It is suggested that the azasteroid may be acting directly on the electron transport system and/or acting indirectly through membrane perturbations which disrupts the electron transport process.

Alanine↗

Effect of fluorophenylalanine on bacteriophage MS2 replication.

A concentration of 10 mug of fluorophenylalanine per ml added to a chemically defined medium reduced by 100-fold the number of bacteriophage MS2 produced on Escherichia coli C3000 and increased the latent period. Fluorophenylalanine was most effective when added concurrent with infection. Addition of a 10-fold greater concentration of phenylalanine reversed the inhibition caused by fluorophenylalanine. Radioactive fluorophenylalanine was incorporated into the coat protein. The four phenylalanine-containing chymotryptic peptides are not equally accessible to fluorophenylalanine. Only two of the peptides are highly labeled by fluorophenylalanine. Incorporation of fluorophenylalanine decreased the specific infectivity and the rate of adsorption but did not increase the sensitivity of the whole virus to ribonuclease. MS2 ribonucleic acid (RNA) functioned as messenger RNA for the incorporation of both phenylalanine and fluorophenylalanine in a cell-free incorporating system from E. coli.

Adsorption↗