Analysis by electron microscopy of the variable segment in the maxi-circle of kinetoplast DNA from Trypanosoma brucei

The 20.5-kbp maxi-circle from the kinetoplast DNA of Trypanosoma brucei contains a 5-kbp segment which is not cut by most restriction endonucleases and which varies in size in closely-related trypanosome strains (Borst, P., Fase-Fowler, F., Hoeijmakers, J.H.J. and Frasch, A.C.C. (1980) Biochim. Biophys. Acta 610, 197–210). We have now analysed partial denaturation maps of the linearized maxi-circles by electron microscopy and find that the variable segment is not more AT-rich than the remainder of the maxi-circle. Early denaturation begins at two separate regions of the maxi-circle outside the variable region and one of these corresponds with the position of the gene for the large (12 S) ribosomal RNA. Denaturation-renaturation of maxi-circles leads to the formation of partially mismatched duplexes that look like underwound loops in electron micrographs. These loops are only found in the variable region and they vary in size and appearance. Under our renaturation conditions single-stranded maxi-circle DNA is devoid of secondary structure and this suggests that the underwound loops arise by misalignment of straight tandem repeats in the DNA. We have also analysed heteroduplexes between maxi-circles from two closely related T. brucei strains that differ by 1 kbp in the size of their variable segment. Most molecules had no underwound loops and contained mismatched regions in the variable segment only. The appearance of these regions is diverse, varying from fully duplex with two single-stranded loops to molecules with a heterogeneous array of smaller loops. The total size of single-stranded DNA in the heteroduplexes may be as high as 1.2 μm, i.e., a factor 4 higher than the size difference between the heteroduplex partners. We conclude that the variable region consists of imperfect tandem repeats of a sequence that evolves rapidly. This region might contain the origin of maxi-circle replication.     

Key parameters controlling stiffness variability within trees: a multiscale experimental–numerical approach

Microstructural properties of wood vary considerably within a tree. Knowledge of these properties and a better understanding of their relationship to the macroscopic mechanical performance of wood are crucial to optimize the yield and economic value of forest stocks. This holds particularly for the end-use requirements in engineering applications. In this study the microstructure–stiffness relationships of Scots pine are examined with a focus on the effects of the microstructural variability on the elastic properties of wood at different length scales. For this purpose, we have augmented microstructural data acquired using SilviScan-3? (namely wood density, cell dimensions, earlywood and latewood proportion, microfibril angle) with local measurements of these quantities and of the chemical composition derived from wide-angle X-ray scattering, light microscopy, and thermogravimetric analysis, respectively. The stiffness properties were determined by means of ultrasonic tests at the clear wood scale and by means of nanoindentation at the cell wall scale. In addition, micro-mechanical modeling was applied to assess the causal relations between structural and mechanical properties and to complement the experimental investigations. Typical variability profiles of microstructural and mechanical properties are shown from pith to bark, across a single growth ring and from earlywood to latewood. The clear increase of the longitudinal stiffness as well as the rather constant transverse stiffness from pith to bark could be explained by the variation in microfibril angle and wood density over the entire radial distance. The dependence of local cell wall stiffness on the local microfibril angle was also demonstrated. However, the local properties did not necessarily follow the trends observed at the macroscopic scale and exhibited only a weak relationship with the macroscopic mechanical properties. While the relationship between silvicultural practice and wood microstructure remains to be modeled using statistical techniques, the influence of microstructural properties on the macroscopic mechanical behavior of wood can now be described by a physical model. The knowledge gained by these investigations and the availability of a new micromechanical model, which allows transferring these findings to non-tested material, will be valuable for wood quality assessment and optimization in timber engineering.     

Preserving Neural Function under Extreme Scaling

Important brain functions need to be conserved throughout organisms of extremely varying sizes. Here we study the scaling properties of an essential component of computation in the brain: the single neuron. We compare morphology and signal propagation of a uniquely identifiable interneuron, the HS cell, in the blowfly (Calliphora) with its exact counterpart in the fruit fly (Drosophila) which is about four times smaller in each dimension. Anatomical features of the HS cell scale isometrically and minimise wiring costs but, by themselves, do not scale to preserve the electrotonic behaviour. However, the membrane properties are set to conserve dendritic as well as axonal delays and attenuation as well as dendritic integration of visual information. In conclusion, the electrotonic structure of a neuron, the HS cell in this case, is surprisingly stable over a wide range of morphological scales.     

A study of the release of phosphate and arsenate from yeast

Under certain conditions yeast cells release previously accumulated phosphate or arsenate. This efflux occurs only when metabolism is not inhibited. There is a rough correlation between the content of orthophosphate and the rate of release.