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BONE CELLS: BIOCHEMICAL AND BIOLOGICAL STUDIES AFTER ENZYMATIC ISOLATION.

Short-term incubation of rat calvaria in buffered crude collagenase permitted the isolation of morphologically intact cells that absorb vital dyes, contain alkaline phosphatase, and multiply in tissue culture. Freshly harvested cells were similar to whole bone segments in aerobic glucose metabolism.

Bone and Bones↗

Enzyme production by species of Cephalosporium.

The culture filtrates of ten species of Cephalosporium, which had been grown under conditions of submerged culture, were tested for enzymatic activity against each of seven substrates. The latter included casein, gelatin, milk, hemoglobin, human plasma clots, starch, and N-acetyl-beta-D-glucosaminide. All organisms tested were active, but to varying degrees. The most pronounced activities were obtained against the proteinaceous substrates. Two unidentified species of Cephalosporium exhibited the highest over-all activities, but no one organism predominated for all enzymatic functions. The ability of a filtrate to degrade a specific substrate was not always correlated with its ability to attack other substrates. The fibrinolytic properties of three of the cephalosporia were of particular interest. alpha-Amylase activity was not significant. The results obtained suggest the possible use of selected species of Cephalosporium as sources of a variety of microbial enzymes.

Acremonium↗

EFFECT OF ENZYMES ON THE INTERACTION OF ENTEROVIRUSES WITH LIVING HELA CELLS.

Zajac, Ihor (Hahnemann Medical College, Philadelphia, Pa.), and Richard L. Crowell. Effect of enzymes on the interaction of enteroviruses with living HeLa cells. J. Bacteriol. 89:574-582. 1965.-Eight crude enzyme preparations and two crystalline enzymes were tested for ability to inactivate coxsackie group B and poliovirus receptors on living HeLa cells. Receptor-destroying enzyme, erepsin, lysozyme, collagenase, proteinase, and cobra venom did not alter attachment of coxsackie B3 or poliovirus T1 to cells, whereas elastase prevented attachment of both viruses tested. Treatment of live cells with pancreatin or chymotrypsin rendered cells unable to attach group B coxsackie viruses, whereas cells treated with trypsin failed to attach poliovirus T1. In addition, chymotrypsin was found to release coxsackie B3 and poliovirus T1 from cell surfaces, whereas trypsin was unable to dissociate virus-cell union. These results indicate that cellular receptors for polioviruses differ from those for group B coxsackie-viruses. The finding that 1% solutions of enzymes will inactivate differentially the enteroviral receptors of HeLa cells, without altering cell viability, provides a useful approach for study of enterovirus receptors of live host cells.

Animals↗