A method for infection of cultured myogenic cells with rous sarcoma virus using polybrene

Summary A method for efficiently infecting primary myogenic cultures with a temperature-sensitive variant of the Prague strain of Rous sarcoma virus (RSV, tsLA24) to obtain a high yield of transformed myogenic cells is reported. It incorporates the use of an amorphous polymer of polycations, Polybrene, to enhance the absorption of the virus by the muscle cells. In addition, other steps which were shown to be important were a) to allow cell attachment before infection, b) to infect at 35° C in low protein medium, c) to use a density of 1 to 1.5×10⁶ cells/60-mm dish, d) gentle agitation during infection, and e) to minimize the number of passages after infection. The use of the temperature-sensitive virus provided a means of confirming the presence of myogenic cells in transformed cultures. When infected cells were maintained at 35° C (the permissive temperature for virus activity) they exhibited the characteristics of transformed cells. These characteristics included altered cell morphology, the absence of contact-inhibited growth, growth in semisolid medium, and expression of thesrc oncogene. In contrast, not expresssrc and showed normal myogenic development and ultimately formed myotubes. This work was supported by grants from the Australian Research Grants Committee and the Cancer Foundation of Western Australia.     

Appearance of the liver form of pyruvate kinase in differentiating cultured foetal hepatocytes

From about the 16th day of gestation three forms of pyruvate kinase are present in foetal rat liver (L, R, and M2). Hepatocytes isolated from 15-day-old foetuses do not possess the liver form of pyruvate kinase, but after three days in culture this enzyme can be detected. No effect on the appearance of the enzyme could be seen by administration of insulin and fructose. Hepatocytes isolated from 19-day-old foetuses exhibit three forms of the enzyme (L, R, and M2) on day 1 of culture but thereafter only two forms are detectable (L and M2). A decrease in activity of the L form is observed. This could be retarded by administration of insulin and fructose.     

Extracellular enzymes of some black aspergilli in Singapore

Summary Ten strains ofAspergillus niger and one strain ofA. awamori showed proteolytic activity and amylolytic and cellulolytic activities were observed in all the strains except inA. niger isolated from rice grains.Aspergillus awamori was shown to be the best producer of amylolytic enzymes but poorest for cellulases. All the strains were shown to be good producers of proteolytic enzymes while the strain from rubber wood seemed to be the best cellulase-producing strain. Both single conidium and hyphal tip inoculation techniques gave similar results. Differences in the levels of extracellular enzymes produced by similar strains were also observed using the two different techniques.

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Enzimas extracelulares de algunos aspergillus negros de Singapur

Resumen Se han aislado diez cepas deAspergillus niger y una deA. awamori que presentan buena actividad proteolítica. Todas las cepas, excepto una deA. niger, aislado de granos de arroz, mostraron, además actividades amilolíticas y celulolíticas. Los mejores productores de enzimas celulolíticas y amilolíticos parecen ser, respectivamente, la cepa deA. niger aislada de madera de caucho yA. awamori. Aspergillus awamori es, sin embargo, el peor productor de celulasas. Se han usado dos técnicas de inoculación, a partir de una solo conidio o bien a partir de un trozo de hifa, obteniéndose resultados comparables. Utilizando estas mismas técnicas se han observado variaciones en las cantidades de enzimas excretados por cepas semejantes.

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Enzymes exo-cellulaires de certains aspergilli noirs isolés à Singapour

Résumé Dix souches d'Aspergillusniger et une d'A.awamori ont été étudiées. Toutes ces souches ont une activité protéolytique et toutes, sauf une souche d'A.niger isolée à partir de grains de riz, ont des activités amylolytiques et cellulolytiques. Il a été constaté qu'Aawamori est le meilleur producteur d'enzymes amylolytiques, mais le moins bon producteur de cellulases. Toutes les sourches sont bonnes productrices d'enzymes protéolytiques, et celle provenant d'un arbre à caoutchouc est la meilleure productrice de cellulase. Les techniques d'inoculation avec une conidie unique ou avec une extrémité d'hyphe donnent des résultats similaires. Les différences de niveau des enzymes exo-cellulaires produits par des souches similaires ont également été observées avec ces deux techniques différentes.

     

Protein content and amino acid composition of cassava leaf

On the basis of leaf dry wt, the protein content of six varieties of cassava varied from 29.3 to 38.6% and the estimated leaf protein production ranged from 242 to 953 kg per ha. On the basis of fr. wt of leaf, the total amino acids ranged from 8.42 to 9.4% while the essential amino acids averaged 4.21% and the sulphur-containing amino acids only 0.25%. The amino acid composition profiles for the six varieties was similar.