The estimation of galactose, mannose and fucose in glycoproteins by radioisotope dilution

1. The principle of radioisotope dilution, as used previously for the estimation of mannose in egg albumin, was applied on a semi-micro scale to the estimation of fucose, mannose and galactose in some glycoproteins. The sugars were separated by partition chromatography on columns of Celite 545. 2. The release of mannose from egg albumin in 2n-hydrochloric acid at 100 degrees after various times was determined by the radioisotope-dilution method and found to have a half-time of 7min. 3. The destruction of mannose in 2n-hydrochloric acid after 3hr. at 100 degrees was found to be small if air was excluded. The destruction was slightly increased by the presence of lysozyme containing tryptophan in an amount equimolar with the mannose. The same amount of free tryptophan caused considerable loss of mannose. 4. Analytical values are reported for the non-amino sugar contents of egg albumin, rabbit gamma-globulin and some samples of blood-group-specific substances. The values found were similar to the most reliable estimates published previously.     

Streptococcus faecium var. casselifavus, nov. var

Streptococcus faecium var. casseliflavus is a gram-positive, spherical cell. The cells occur chiefly as pairs within chains and elongate to ogive-shaped cells during growth. Growth is good on 5% bile salts-agar and in broth at 10 C, and in broth adjusted to pH 9.6 or containing 6.5% NaCl, but many strains fail to grow at 45 C. Litmus is reduced rapidly prior to formation of an acid curd. Few strains release ammonia from arginine or serine. The organism is not proteolytic and does not produce H(2)S or acetylmethylcarbinol, reduce nitrate, decarboxylate tyrosine, or produce slime on sucrose-agar. Most strains survive heating to 60 C for 30 min. It produces gray colonies on potassium tellurite agar, reduces 2,3,5-triphenyltetrazolium-HCl to a pink color, and ferments cellobiose, dextrin, maltose, mannose, and sorbitol, thus resembling S. faecalis. Like S. faecium, it produces peroxidase but not catalase on heated blood media, dissimilates malate, and ferments arabinose, melibiose, and salicin, but not melezitose. Like both species, it ferments dextrose, galactose, lactose, mannitol, sucrose, trehalose, and citrate. Properties peculiar to the variant include the high pH limiting initiation and termination of growth; the fermentation of alpha-methyl-d-glucoside, raffinose, and xylose; motility; and growth without blue button formation in ethyl violet broth. The water-soluble, pale lemon-yellow pigment is released into the aqueous phase only after the cell envelope is altered by fat solvents. The bacterium thrives as an epiphyte on plants.     

Selective Inhibition of Reovirus Ribonucleic Acid Synthesis by Cycloheximide

Cycloheximide, at a concentration of 10 mug/ml, rapidly blocked protein synthesis in L cells infected with reovirus. When the drug was added before 5 hr postinfection, synthesis of both single- and double-stranded varieties of virus-specific ribonucleic acid (RNA), which normally commences between 6 and 7 hr after infection, was blocked. When the cycloheximide was added at 9 hr after infection, uptake of uridine-H(3) into RNA, for the succeeding 6 hr at least, was similar to that of an infected culture without the drug. This latter uptake of uridine-H(3) in the presence of cycloheximide was largely into single-stranded RNA, since double-stranded RNA synthesis was rapidly and markedly inhibited by the cycloheximide. Single-stranded RNA formed in the presence of cycloheximide was found not to be a precursor of viral progeny, double-stranded RNA. Synthesis of double-stranded RNA in the infected cell probably requires prior synthesis of a new protein, which has a rapid rate of turnover. The possibility that formation of single-stranded RNA is preceded by synthesis of a second new protein is discussed.