Guanine nucleotide contacts within viral DNA sequences bound by polyomavirus large T antigen.

Essential nucleotide contacts between the polyomavirus large T antigen and its multiple specific binding regions within the regulatory sequences of the polyomavirus genome were determined in vitro by methylation interference. Methylation of any of the guanine residues of the 5'-G(A/G)GGC-3' pentanucleotide repeats in large-T-antigen-binding regions A, B, C, and 3 (A. Cowie and R. Kamen, J. Virol. 52:750-760, 1984) interfered with T antigen binding. Within regions A, B, and C these pentanucleotides are spaced 5 or 6 base pairs apart. Therefore, the clusters of contacted nucleotides within each of these binding regions are localized along one face of the DNA helix. Methylation of guanines within the sequences between the pentanucleotide repeats did not interfere with binding. The ORI binding region contains four additional pentanucleotide sequences within a region of dyad symmetry. Methylation of only particular guanines of these pentanucleotides interfered with T antigen binding. The spatial arrangement of the pentanucleotides in the ORI is such that the clusters of contacted guanines are situated around the DNA helix, thereby forming a very different arrangement from that found in the other binding regions. A model is discussed in which cooperative interactions between T antigen protomers, recognizing individual pentanucleotides, determines the strength and the function of different T antigen-DNA interactions.     

Lack of correlation between differentiation status and response to radiation of three murine squamous cell carcinomas

Summary The gross growth rate, histology, cellular kinetics, andin situ radiobiological response have been measured for three murine, keratinising squamous cell carcinomas that differed in their degree of differentiation. Growth rate was fastest in the least-differentiated tumour, slowest in the best-differentiated. However, the kinetics of the compartment of undifferentiated cells that are likely to be radiotherapeutically important, were the same for the three lines. There was no correlation between degree of differentiation and intrinsic or apparent radiosensitivity as measured by the growth delay assay. The radiobiologically best-oxygenated tumour was that which had the largest stromal component and this was not the best-differentiated tumour.     

A simple and reliable turbidimetric and kinetic assay for alpha-amylase that is readily applied to culture supernatants and cell extracts

Summary A simple, reliable and sensitive assay for alpha-amylase activity is reported, together with its theoretical derivation, that overcomes many of the problems encountered with other assays, especially when attempting to assay alpha-amylase activity in crude cell extracts or culture supernatants. The method relies on the reduction in turbidity that occurs upon digestion of a starch suspension with alpha-amylase. The initial rate of decrease in turbidity is shown to be proportional to a wide range of enzyme concentrations, permitting a rapid spectrophotometric and kinetic determination of alpha-amylase activity.     

Effects of waterlogging on chickpeas II. Possible causes of decreased tolerance of waterlogging at flowering

Waterlogging tolerance of chickpeas was found, in earlier work, to decrease sharply at flowering. Three experiments were performed to attempt to explain the mechanisms involved in this response. In the first, a range of treatments was imposed to modify the plant's source/sink relationships, as carbohydrate supply and partitioning were considered possible determinants of waterlogging tolerance. Plants from which buds were removed showed the most rapid recovery after waterlogging. Defoliation immediately before waterlogging reduced the rate of recovery. Application of benzyladenine plus gibberellic acid prior to waterlogging delayed stomatal closure and leaf senescence, inhibited apical growth and stimulated axillary growth. The second experiment aimed to confirm the influence of bud removal and to determine whether waterlogging tolerance is correlated with carbohydrate supply. Treatments comprised two sowing times, ten days apart, and two bud treatments (retained and removed). Waterlogging was imposed when older plants had been flowering for seven days and younger plants were in bud. Waterlogging caused soluble sugars to accumulate in the lower stem, suggesting that a deficiency of assimilates did not contribute to waterlogging injury. Similarly, waterlogging increased nitrogen concentration in the stem, through mobilisation from senescing leaves. Bud removal enhanced leaf survival and reduced mortality rate after waterlogging; it also increased starch concentration in the lower stem, indicating that storage of assimilates decreased in flowering plants. However, across all treatments, starch concentration was not correlated with waterlogging tolerance. In the third experiment, the effect of the senescence-promoting factor ethylene on preflowering and flowering plants was assessed, using the ethylene-releasing agent ethephon. Ethephon reduced growth to a slightly greater extent when applied prior to flowering than at flowering. There was no evidence that inadequate supply of carbohydrates or nitrogen in the stem, or increased sensitivity to ethylene, contributed to waterlogging intolerance in flowering chickpea plants.     

The Effects of Early Feeding Experience on Long-term Seed Choice by Canaries (Serinus canaria)

Early experience has been found to have long-term effects on behaviour such as social imprinting and song learning. In this paper we investigate whether early experience with particular types of food can lead to long-term feeding preferences in canaries. For 8 wk following the onset of independent foraging, two groups of naive fledgling canaries were reared on different diets. One group had a choice of four seed types, while individuals in the other group received only one type of seed. After this training period, all the birds were given a mixed seed diet for a further 15 wk, and then their preference for the original seeds was tested. All six birds that had been given four seeds during the training period showed the same preference, namely: hemp > niger > millet > linseed. However, all the birds that had been trained on one seed type only, still preferred the seed on which they had been trained. After a further 23 wk on a mixed-seed diet, birds from the restricted early diet group showed no further preference for the seed on which they were trained, and their seed choice at this stage was similar to the birds that had received four seeds. We conclude that early experience can affect food choice in canaries for at least 15 wk, but is subordinate to other influences in the longer term.