Firefly luciferase as a reporter of regulated gene expression in higher plants

The firefly luciferase, assayedin vivo with a low-light video camera, acts as a non-invasive, real-time reporter of the temporal and spatial regulation of gene expression in single plants. Furthermore, the sensitivity of the luciferase assay in extracts of transformed plant tissue makes it a particularly useful marker in transient or stable transformation experiments.     

Unusual Microbial Xylanases from Insect Guts

Recombinant DNA technologies enable the direct isolation and expression of novel genes from biotopes containing complex consortia of uncultured microorganisms. In this study, genomic libraries were constructed from microbial DNA isolated from insect intestinal tracts from the orders Isoptera (termites) and Lepidoptera (moths). Using a targeted functional assay, these environmental DNA libraries were screened for genes that encode proteins with xylanase activity. Several novel xylanase enzymes with unusual primary sequences and novel domains of unknown function were discovered. Phylogenetic analysis demonstrated remarkable distance between the sequences of these enzymes and other known xylanases. Biochemical analysis confirmed that these enzymes are true xylanases, which catalyze the hydrolysis of a variety of substituted β-1,4-linked xylose oligomeric and polymeric substrates and produce unique hydrolysis products. From detailed polyacrylamide carbohydrate electrophoresis analysis of substrate cleavage patterns, the xylan polymer binding sites of these enzymes are proposed.     

Development rates,fecundity and survival of developmental stages of the ticksRhipicephalus appendiculatus,Boophilus decoloratus andB. microplus under field conditions in Zimbabwe

To determine development rates, fecundity and survival ofRhipicephalus appendiculatus, Boophilus decoloratus andB. microplus, a study was carried out in long and short grass in the highveld of Zimbabwe. Engorged adult females of the three species and engorged larvae and nymphs ofR. appendiculatus were buried beneath the soil in small cages in the rainy, cool and hot seasons in 1980 and 1981. Half the number of cages were examined regularly to determine development rates and half were left undisturbed to determine survival rates and the fecundity of engorged females. Development was most rapid during warm conditions and slowest during cool conditions, but high temperatures appeared to prolong the preoviposition periods of all species. The relationship between fluctuating temperatures and rate of development in the field was defined using a least-squares procedure. Survival of engorged females was usually high, but was reduced by predation when they were not protected. Fecundity was reduced in long grass during the cool season and in short grass during the hot season. A higher percentage of eggs hatched in the rainy season than in the cool or dry seasons. The survival of engorged larvae and nymphs was usually high in all seasons. Engorged nymphs were the hardiest stage and eggs the most suceptible stage to adverse microclimatic conditions.     

Computed-tomography-based finite-element models of long bones can accurately capture strain response to bending and torsion

Finite element (FE) models of long bones constructed from computed-tomography (CT) data are emerging as an invaluable tool in the field of bone biomechanics. However, the performance of such FE models is highly dependent on the accurate capture of geometry and appropriate assignment of material properties. In this study, a combined numerical-experimental study is performed comparing FE-predicted surface strains with strain-gauge measurements. Thirty-six major, cadaveric, long bones (humerus, radius, femur and tibia), which cover a wide range of bone sizes, were tested under three-point bending and torsion. The FE models were constructed from trans-axial volumetric CT scans, and the segmented bone images were corrected for partial-volume effects. The material properties (Young's modulus for cortex, density-modulus relationship for trabecular bone and Poisson's ratio) were calibrated by minimizing the error between experiments and simulations among all bones. The R(2) values of the measured strains versus load under three-point bending and torsion were 0.96-0.99 and 0.61-0.99, respectively, for all bones in our dataset. The errors of the calculated FE strains in comparison to those measured using strain gauges in the mechanical tests ranged from -6% to 7% under bending and from -37% to 19% under torsion. The observation of comparatively low errors and high correlations between the FE-predicted strains and the experimental strains, across the various types of bones and loading conditions (bending and torsion), validates our approach to bone segmentation and our choice of material properties.