Coated Vesicles from the Protozoan Parasite Trypanosoma brucei: Purification and Characterization

ABSTRACT. A procedure was developed to purify a coated vesicle fraction from the protozoan parasite Trypanosoma brucei. Electron microscopy revealed a difference between T. brucei coated vesicles and clathrin-coated vesicles from other eukaryotes: trypanosome vesicles were larger (100 to ISO nm in diameter) and contained an inner coat of electron-dense material in addition to the external coat. Evidence suggests that the internal coat is the parasite's variant surface glycoprotein (VSG) coat. The SDS-PAGE analysis shows the major protein of T. brucei coated vesicles has a molecular mass of 61 kD, similar to VSG; this protein was recognized in an immunoblot by anti-VSG serum. Trypanosome coated vesicles also contain a protein which comigrates with the major protein (clathrin) of coated vesicles purified from rat brains. However, this protein is a minor component and it is not serologically cross-reactive with mammalian clathrin. Immunoblot analysis demonstrated that the parasite vesicles contained host IgG, IgM, and serum albumin.     

The Effect of Temperature on Photosynthesis and Carbon Fluxes in Sunflower and Rape

Sunflower (Helianthus annuusL.) and oilseed rape (Brassica napusL.) were grown at constant temperatures of 30 ?C (warm) and13 ?C (cold). Maximal rates of photosynthesis between 5 ?C and35 ?C were at higher temperatures in sunflower than rape. Photosyntheticrate over 4 h at the growth temperature declined in warm-andcold-grown rape and cold-grown sunflower, but remained constantin warm-grown sunflower. The stimulation of photosynthesis by2.0 kPa O₂ compared to 21 kPa O₂ declined with decreasing temperature.At 10 ?C in warm-grown rape photosynthesis was insensitive to2.0 kPa O₂. However, sensitivity to low O₂ continued at 10 ?Cin warm-grown sunflower. Carbohydrates accumulated in the cold,particularly fructose, glucose and sucrose in warm-grown sunflowertransferred to 13 ?C. By monitoring changes of ¹⁴C in leaves after the assimilationof ¹⁴CO₂, the rates of carbon export from leaves, pool sizesand carbon fluxes between them were estimated. The transferof warm- and cold-grown rape to 13 ?C and 30 ?C, respectively,had little effect on these parameters over 22 h. However, exportof carbon from sunflower leaves at 13 ?C was markedly less thanat 30 ?C, irrespective of the growth temperature, due to slowerexport from the transport pool. The rapid suppression of carbonexport at 13 ?C in warm-grown sunflower may be due to inhibitedtranslocation rather than reduced sink demand in the cold. It is concluded that assimilate utilisation is more depressedin the cold than is photosynthesis; this imposes a greater restrictionon biomass production in sunflower than in rape. Key words: Sunflower, rape, temperature, photosynthesis, carbon fluxes     

Changes in phosphorus and nitrogen cycling following food web manipulations in a shallow Dutch lake

1. The flow of phosphorus and nitrogen through the food web of the shallow, eutrophic lake Wolderwijd was analysed for 2 different years before and for 1 year after food web manipulation.
2. After fish removal the water became clear and the growth of macrophytes began. Fish removal resulted in a significant reduction of the total nutrient pool in the water, but differences between the nutrient cycles before and after the experiment were mainly caused by a gradual change driven by a reduced phosphorus input.
3. The zooplankton biomass before and after food web manipulation did not change significantly. Unfavourable food conditions and predation by young fish limited zooplankton biomass after the food web manipulation.
4. After fish removal benthic algae, fish, zoobenthos and macrophytes form the largest pools of nutrients apart from the sediment top layer. However, they contribute only little to nutrient cycles in the water column.     

Endemism and ecological islands: the ostracods from Jamaican bromeliads

1. The inhabitants of ecological islands, such as the phytolemata of bromeliad plants, provide an opportunity to examine the genetic patterns resulting from island radiations and draw inferences about modes of speciation.
2. Allozyme electrophoresis, as well as mitochondrial DNA and morphological analyses, were employed to delimit species boundaries and assess the population genetic structure of the ostracods from bromeliads on the island of Jamaica.
3. Nine species from the genus Elpidium were both sexually reproducing and endemic to the island. The other commonly encountered ostracod genus, Candonopsis , was represented by at least two sexually reproducing species.
4. Most Elpidium species showed very restricted distributions, low heterozygosity, and marked gene pool fragmentation, suggesting that population bottlenecks have occurred frequently. Whether founder events played a causal role in the diversification of these ostracods remains uncertain, but bromeliad habitats seem to be a source of high biodiversity, and may represent an ideal setting for extensive allopatric speciation.     

Collembolan water relations and environmental change in the maritime Antarctic

The water status of the collembolan Cryptopygtus antarcticus (Willem) was investigated from April 1984 to December 1987 at Signy Island, maritime Antarctic, by monthly field sampling to determine body water content. Water content, expressed either as the weight of water per unit dry weight or as a proportion of fresh weight, exhibited both a seasonal cycle and an upward trend over the 44-month study, both of which were highly significant. On an annual basis, body water content was at a minimum (1.21 g g^?1) in July and maximal (1.98 g g^?1) in September, whilst over the entire study water contents increased from 1.3 to 2.0 g g^?1 (or 57-66% of fresh weight) calculated from the fitted linear regression line. Field water contents were below those found for this species in culture (2.9-5.9 g g^?1). Individual C. antarcticus survived experimental loss of 20% of their body water with a resultant significant rise in haemolymph osmolarity from 285 to 397 mOsm L^?1 and there was no evidence of osmoregulation under the experimental conditions of 20 °C and 35% relative humidity. The cuticular permeability (mean conductance) of individual Collembola in dry air increased exponentially with temperature over the range D-45 °C (Q₁₀= 2.0) showing no control of water loss. The physiological response of C. antarcticus suggests that it experiences water stress in its maritime Antarctic habitats with significant seasonal variations of body water content, which correlate with annual cycles of water availability. It is concluded that the significant rise in its mean body water content over the 44-month field study was associated with increased glacial ablation due to higher levels of irradiation and windspeed making available more liquid water. Analyses of climate records for Signy Island from 1947 to 1990 showed that mean monthly air temperature rose by 0.93 °C over this period and by 2.29 °C during the 1980s, both statistically significant increases. Mean monthly windspeeds also increased significantly during 1970–90, and it is suggested that this parameter is the primary climatic driving force behind the increase in glacial ablation during the last two decades. The field water status of species such as C. antarcticus may reflect changes in the patterns of atmospheric circulation, associated with the circumpolar vortex, through increased ozone depletion due to increased tropospheric concentrations of halocarbons.     

The lepidopteran mitochondrial control region: structure and evolution

For several species of lepidoptera, most of the approximately 350-bp mitochondrial control-region sequences were determined. Six of these species are in one genus, Jalmenus; are closely related; and are believed to have undergone recent rapid speciation. Recent speciation was supported by the observation of low interspecific sequence divergence. Thus, no useful phylogeny could be constructed for the genus. Despite a surprising conservation of control-region length, there was little conservation of primary sequences either among the three lepidopteran genera or between lepidoptera and Drosophila. Analysis of secondary structure indicated only one possible feature in common--inferred stem loops with higher-than-random folding energies-- although the positions of the structures in different species were unrelated to regions of primary sequence similarity. We suggest that the conserved, short length of control regions is related to the observed lack of heteroplasmy in lepidopteran mitochondrial genomes. In addition, determination of flanking sequences for one Jalmenus species indicated (i) only weak support for the available model of insect 12S rRNA structure and (ii) that tRNA translocation is a frequent event in the evolution of insect mitochondrial genomes.