Ultraviolet Inactivation and Photoproducts of Transforming DNA Irradiated at Low Temperatures

Solutions of Haemophilus influenzae transforming DNA were irradiated at temperatures ranging from 25°C to - 196°C. Temperature dependence of the formation of thymine-containing dimers was closely correlated with inactivation of transforming activity; in general, both dimerization and inactivation decreased with decreasing temperature. The fraction of nonphotoreactivable damage increased with increasing dose at low temperatures. The nonphotoreactivable spore-type photoproduct was formed at low temperatures with a maximum at - 100°C, a temperature at which the nonphotoreactivable biological inactivation was also a maximum. Intrastrand cross-linking, like dimer formation, decreased with decreasing irradiation temperature.     

The conversion of injected glucose into renal glycogen and mucopolysaccharides

Summary Tritiated glucose has been injected into rabbits in various states of hydration. The renal papilla of all animals showed an uptake of the label, converted into glycogen, and into mucopolysaccharides, in a manner dependent on the water balance of the animal.In papillae of control animals, the glycogen of the collecting duct epithelial cells and the mucopolysaccharides of the interstitium were labelled.In papillae of animals in an aqueous diuresis, the collecting duct glycogen was lightly labelled and there was no label over the interstitium.Antidiuretic hormone caused a diversion of label from the collecting ducts into interstitial mucopolysaccharides.The significance of these findings, with respect to renal concentrating ability, is discussed.The author wishes to thank Dr. M. K. S. Hathorn for help with the statistical analysis. Mr. K. Gamblin and Mr. P. L. Hyam gave valuable technical assistance. This research formed a part of the work approved for the degree of Ph. D. (London).     

Purification and characterization of beta-glucosidase of Alcaligenes faecalis

A cellobiose-utilizing bacterium isolated from sugar cane bagasse and identified as a strain of Alcaligenes faecalis (ATCC 21400) produced an inducible beta-glucoside-splitting enzyme. The enzyme was purified by a series of streptomycin and ammonium sulfate fractionations and by Sephadex and diethylaminoethyl column chromatography. The final preparation was purified 130-fold, with a recovery of about 10% of the initial enzyme activity. The enzyme had a wide pH range, with optimal activity at pH 6.0 to 7.0. The enzyme was stable in solution at pH 6.5 to 7.8 when kept at 30 C for 2 hr, but it was destroyed by temperatures above 55 C. At 58 and 60 C, the time required to inactivate 90% of the initial activity was 16 and 6.5 min, respectively. An activation energy of 9,500 cal/mole and a K(m) of 1.25 x 10(-4)m were obtained by using p-nitrophenyl beta-glucoside as a substrate. The K(i) value and hydrolysis of cellobiose by the enzyme indicated a high affinity of the enzyme for the cellobiose. The enzyme had its specificity on beta-glucosidic linkage and the rate of hydrolisis of glucosides depended upon the nature of the aglycon moiety. The inactivation studies showed the presence of sulfhydryl groups in the enzyme. The activity of the enzyme was easily destroyed by the Cu(++) and Hg(++) ions. The Michaelis-Menton relationship and the rate of heat inactivation indicated the presence of one type of noninteracting active site in the bacterial beta-glucosidase. Molecular weight of the enzyme was estimated by gel filtration (Sephadex G-200) and sucrose density gradient, and a value of 120,000 to 160,000 was obtained.     

Isolation and Characterization of a Cellulose-utilizing Bacterium

A cellulose-decomposing aerobic and mesophilic bacterium has been isolated from soils of sugar cane fields. The terminal dilution method was adapted to isolate a single clone of cellulolytic organism from closely related contaminants. The cultural and physiological characteristics of the isolate were studied, and the organism was identified as a member of the genus Cellulomonas. The isolate excreted cellulase into the menstruum, and it hydrolyzed various cellulosic materials producing cellobiose as the final breakdown product in the menstruum. When sugar cane bagasse was properly treated with alkali and heat, the organism could decompose up to 90% of the initial substrate within 5 days. Amino acid analysis of the cell crop revealed a high content of lysine, and the essential amino acid pattern compared favorably with that of Food and Agricultural Organization reference protein.     

Distribution of hereditary blood groups among Indians in South America. IV. In Chile with inferences concerning genetic connections between Polynesia and America

This is the fourth paper in a series on the distribution of blood groups among Indians of South America. It reports the findings on the Indians of Chile and the Polynesians of Chile's Easter Island. Blood specimens were procured from the following putatively pure Indians and unmixed Polynesians: 44 Alacaluf of Puerto Eden, Isla Wellington, 141 Mapuche (Araucanian) of Lonquimay, Malleco Province, 80 Atacameños of Antofagasta Province, and 45 Polynesians of Easter Island. These 310 samples were tested for blood factors in the A-B-O, M-N-S-s, P, Rh-Hr, Lutheran, K-k, Lewis, Duffy, Kidd and Diego systems, and for the Wright (Wr^a) agglutinogen. Serum samples were tested for haptoglobins and transferrins. Hemolysates prepared from the blood clots were tested for hemoglobin types. The results are presented as phenotype incidences and calculated gene frequencies in appropriate tables. Locations of the populations from which blood samples were procured are shown on two maps. The high frequencies for the O gene usually reported for South American Indians obtain in putatively pure Chilean Indians but A¹ is high in Easter Island Polynesians. In both Indians and Polynesians M, s, R¹ (CDe), R² (cDE), Lu^b, k, Le^H, and Fy^a gene frequencies are high and B, N, S, Mi^a, Vw, Rº (cDe), r (cde), Lu^a, K, Le¹, Fy^b, and Wr^a (Ca) are low or absent. The Diego (Di) gene is present in the Mapuche and Atacameños but absent in the Alacaluf and Polynesians. Hp¹ gene frequencies were determined only in the Alacaluf and Atacameños, in which they are 0.48 and 0.67 respectively. Transferrins were determined for the Alacaluf and Atacameños Indians and all were classified as Tf C. All Chilean Indian and Polynesian specimens were tested electrophoretically for hemoglobin types and all contained only hemoglobin (A) as a major component.