Vitamin B12 stalls the 80 S ribosomal complex on the hepatitis C internal ribosome entry site

The effect of cyanocobalamin (CNCbl, vitamin 1312) on hepatitis C virus internal ribosome entry site (HCV IRES)-dependent initiation of translation was studied by ribosomal toeprinting and sucrose gradient centrifugation analysis. These results suggested that CNCbl did not inhibit HCV IRES-dependent translation by a competitive binding mechanism. CNCbl allowed 80 S elongation complex formation on the mRNA, but stalled the initiation at that point, effectively trapping the 80 S ribosomal complexes on the HCV TRES. CNCbl had no effect on cap-dependent mRNA, consistent with the known mRNA specificity of this translational inhibitor. To help elucidate the mechanism, comparative data were collected for the well-characterised translation inhibitors cycloheximide and 5'-guanylyl-imidophosphate, Although CNCbl stalled HCV IRES-dependent translation at approximately the same step in initiation as cycloheximide, the mechanisms of these two inhibitors are distinct. (C) 2002 Elsevier Science Ltd. All rights reserved.

Development of a fully quantitative approach to the interpretation of stress echocardiography using radial and longitudinal myocardial velocities

Background: Tissue Doppler may be used to quantify regional left ventricular function but is limited by segmental variation of longitudinal velocity from base to apex and free to septal walls. We sought to overcome this by developing a composite of longitudinal and radial velocities. Methods and Results. We examined 82 unselected patients undergoing a standard dobutamine echocardiogram. Longitudinal velocity was obtained in the basal and mid segments of each wall using tissue Doppler in the apical views. Radial velocities were derived in the same segments using an automated border detection system and centerline method with regional chords grouped according to segment location and temporally averaged. In 25 patients at low probability of coronary disease, the pattern of regional variation in longitudinal velocity (higher in the septum) was the opposite of radial velocity (higher in the free wall) and the combination was homogenous. In 57 patients undergoing angiography, velocity in abnormal segments was less than normal segments using longitudinal (6.0 +/- 3.6 vs 9.0 +/- 2.2 cm/s, P = .01) and radial velocity (6.0 +/- 4.0 vs 8.0 +/- 3.9 cm/s, P = .02). However, the composite velocity permitted better separation of abnormal and normal segments (13.3 +/- 5.6 vs 17.5 +/- 4.2 cm/s, P = .001). There was no significant difference between the accuracy of this quantitative approach and expert visual wall motion analysis (81% vs 84%, P = .56). Conclusion: Regional variation of uni-dimensional myocardial velocities necessitates site-specific normal ranges, probably because of different fiber directions. Combined analysis of longitudinal and radial velocities allows the derivation of a composite velocity, which is homogenous in all segments and may allow better separation of normal and abnormal myocardium.

Vertical fluxes of sediment in oscillatory sheet flow

Time series of vertical sediment fluxes are derived from concentration time series in sheet flow under waves. While the concentrations C(z,t) vary very little with time for \z\ < 10d(50), the measured vertical sediment fluxes Q(zs)(z,t) vary strongly with time in this vertical band and their time variation follows, to some extent, the variation of the grain roughness Shields parameter 02,5(t). Thus, sediment distribution models based on the pickup function boundary condition are in some qualitative agreement with the measurements. However, the pickup function models are only able to model the upward bursts of sediment during the accelerating phases of the flow. They are, so far, unable to model the following strong downward sediment fluxes, which are observed during the periods of flow deceleration. Classical pickup functions, which essentially depend on the Shields parameter, are also incapable of modelling the secondary entrainment fluxes, which sometimes occur at free stream velocity reversal. The measured vertical fluxes indicate that the effective sediment settling velocity in the high [(0.3 < C(z,t) < 0.4] concentration area is typically only a few percent of the clear water settling velocity, while the measurements of Richardson and Jeronimo [Chem. Eng. Sci. 34 (1979) 1419], from a different physical setting, lead to estimates of the order 20%. The data does not support gradient diffusion as a model for sediment entrainment from the bed. That is, detailed modelling of the observed near-bed fluxes would require diffusivities that go negative during periods of flow deceleration. An observed general trend for concentration variability to increase with elevation close to the bed is also irreconcilable with diffusion models driven by a bottom boundary condition. (C) 2002 Published by Elsevier Science B.V.

Investigation of the flow field of a highly heated jet of air

Measurements of mean and fluctuating velocity and temperature and their self- and cross-products to the third-order are presented for a heated axisymmetric air jet. Froude numbers in the range of 3500 13,190, Reynolds numbers in the range of 3470-8500 and non-dimensional streamwise distances. X*, from 0.27 to 1.98 are covered by the data. It was found that turbulence intensity decreases for the heated jet in the region between the inertia dominated and the buoyancy dominated regions which is contrary to findings with helium jets mixing with ambient air to produce density fluctuations. The effects of heating on the turbulent kinetic energy budget and the temperature variance budget show small differences for the inertia dominated region and the intermediate region which help to explain the transition process to the far field plume region. Constants are evaluated for the isotropic eddy diffusivity and generalised gradient hypothesis models as well as the scalar variance model. No significant effect of heating on the dissipation time-scale ratio was found. A novel wire array with an inclined cold wire was used. Measurements obtained with this probe are found to lead to asymmetries in some of the higher-order products. Further investigation suggested that the asymmetries are attributable to an as yet unreported interference effect produced by the leading prong of the inclined temperature wire, The effect may also have implications for inclined velocity wires which contain a temperature component when used in heated flows. (C) 2002 Elsevier Science Inc. All rights reserved.

Analysis of the wind field and heat budget in an alpine lake basin during summertime fair weather conditions

Observational data collected in the Lake Tekapo hydro catchment of the Southern Alps in New Zealand are used to analyse the wind and temperature fields in the alpine lake basin during summertime fair weather conditions. Measurements from surface stations, pilot balloon and tethersonde soundings, Doppler sodar and an instrumented light aircraft provide evidence of multi-scale interacting wind systems, ranging from microscale slope winds to mesoscale coast-to-basin flows. Thermal forcing of the winds occurred due to differential heating as a consequence of orography and heterogeneous surface features, which is quantified by heat budget and pressure field analysis. The daytime vertical temperature structure was characterised by distinct layering. Features of particular interest are the formation of thermal internal boundary layers due to the lake-land discontinuity and the development of elevated mixed layers. The latter were generated by advective heating from the basin and valley sidewalls by slope winds and by a superimposed valley wind blowing from the basin over Lake Tekapo and up the tributary Godley Valley. Daytime heating in the basin and its tributary valleys caused the development of a strong horizontal temperature gradient between the basin atmosphere and that over the surrounding landscape, and hence the development of a mesoscale heat low over the basin. After noon, air from outside the basin started flowing over mountain saddles into the basin causing cooling in the lowest layers, whereas at ridge top height the horizontal air temperature gradient between inside and outside the basin continued to increase. In the early evening, a more massive intrusion of cold air caused rapid cooling and a transition to a rather uniform slightly stable stratification up to about 2000 m agl. The onset time of this rapid cooling varied about 1-2 h between observation sites and was probably triggered by the decay of up-slope winds inside the basin, which previously countered the intrusion of air over the surrounding ridges. The intrusion of air from outside the basin continued until about mid-night, when a northerly mountain wind from the Godley Valley became dominant. The results illustrate the extreme complexity that can be caused by the operation of thermal forcing processes at a wide range of spatial scales.

Inferring genome trees by using a filter to eliminate phylogenetically discordant sequences and a distance matrix based on mean normalized BLASTP scores

Darwin's paradigm holds that the diversity of present-day organisms has arisen via a process of genetic descent with modification, as on a bifurcating tree. Evidence is accumulating that genes are sometimes transferred not along lineages but rather across lineages. To the extent that this is so, Darwin's paradigm can apply only imperfectly to genomes, potentially complicating or perhaps undermining attempts to reconstruct historical relationships among genomes (i.e., a genome tree). Whether most genes in a genome have arisen via treelike (vertical) descent or by lateral transfer across lineages can be tested if enough complete genome sequences are used. We define a phylogenetically discordant sequence (PDS) as an open reading frame (ORF) that exhibits patterns of similarity relationships statistically distinguishable from those of most other ORFs in the same genome. PDSs represent between 6.0 and 16.8% (mean, 10.8%) of the analyzable ORFs in the genomes of 28 bacteria, eight archaea, and one eukaryote (Saccharomyces cerevisiae). In this study we developed and assessed a distance-based approach, based on mean pairwise sequence similarity, for generating genome trees. Exclusion of PDSs improved bootstrap support for basal nodes but altered few topological features, indicating that there is little systematic bias among PDSs. Many but not all features of the genome tree from which PDSs were excluded are consistent with the 16S rRNA tree.

Multiple ciguatoxins present in Indian Ocean reef fish

Optimised gradient reversed-phase high-performance liquid chromatography electrospray ionisation mass spectrometry (LC/MS) methods, in combination with a [H-3]-brevetoxin binding assay (RLB), revealed multiple ciguatoxins in a partially purified extract of a highly toxic Lutjanus sebae (red emperor) from the Indian Ocean. Two major ciguatoxins of 1140.6 Da (I-CTX-1 and -2) and two minor ciguatoxins of 1156.6 Da (I-CTX-3 and -4) were identified. Accurate mass analysis revealed that I-CTX-1 and -2 and Caribbean C-CTX-1 had indistinguishable masses (1140.6316 Da, at 0.44 ppm resolution). Toxicity estimated from LC/MS/RLB responses indicated that I-CTX-1 and -2 were both similar to 60% the potency of Pacific ciguatoxin-1 (P-CTX-1). In contrast to ciguatoxins of the Pacific where the more oxidised ciguatoxins are more potent, I-CTX-3 and -4 were similar to 20% of P-CTX-1 potency. Interconversion in dilute acid or on storage, typical of spiroketal and hemiketal functionality found in P-CTXs and C-CTXs, respectively, was not observed to occur between I-CTX-1 and -2. The ratio of CTX-1 and -2 varied depending on the fish extract being analysed. These results suggest that I-CTX-1 and -2 may arise from separate dinoflagellate precursors that may be oxidatively biotransformed to I-CTX-3 and -4 in fish. (C) 2002 Published by Elsevier Science Ltd.

The prediction of sharkskin instability observed during film blowing

The characteristics of sharkskin surface instability for linear low density polyethylene are studied as a function of film blowing processing conditions. By means of scanning electron microscopy and surface profilometry, is it found that for the standard industrial die geometry studied, sharkskin only occurs on the inside of the film bubble. Previous work suggests that this instability may be due to critical extensional stress levels at the exit of the die. Isothermal integral viscoelastic simulations of the annular extrusion process are reported, and confirm that the extensional stress at the die exit is large enough to cause local melt rupture. However the extensional stress level at the outer die wall predicts melt rupture of the outside bubble surface also, which contradicts the experimental findings. A significant temperature gradient is expected to exist across the die gap at the exit of the die, due to the external heating of the die and the low conductivity, of the polymer melt. It is shown that a gradient of 20 degreesC is required to cause sharkskin to only appear on the inner bubble surface.

Finite difference time domain (FDTD) method for modeling the effect of switched gradients on the human body in MRI

Numerical modeling of the eddy currents induced in the human body by the pulsed field gradients in MRI presents a difficult computational problem. It requires an efficient and accurate computational method for high spatial resolution analyses with a relatively low input frequency. In this article, a new technique is described which allows the finite difference time domain (FDTD) method to be efficiently applied over a very large frequency range, including low frequencies. This is not the case in conventional FDTD-based methods. A method of implementing streamline gradients in FDTD is presented, as well as comparative analyses which show that the correct source injection in the FDTD simulation plays a crucial rule in obtaining accurate solutions. In particular, making use of the derivative of the input source waveform is shown to provide distinct benefits in accuracy over direct source injection. In the method, no alterations to the properties of either the source or the transmission media are required. The method is essentially frequency independent and the source injection method has been verified against examples with analytical solutions. Results are presented showing the spatial distribution of gradient-induced electric fields and eddy currents in a complete body model.

Modelling activated sludge flocculation using population balances

Activated sludge flocculation was modelled using population balances. The model followed the dynamics of activated sludge flocculation providing a good approximation of the change in mean floe size with time. Increasing the average velocity gradient decreased the final floe size. The breakage rate coefficient and collision efficiency also varied with the average velocity gradient. A power law relationship was found for the increase in breakage rate coefficient with increasing average velocity gradient. Further investigation will be conducted to determine the relationship between the collision efficiency and particle size to provide a better approximation of dynamic changes in the floe size distribution during flocculation. (C) 2002 Published by Elsevier Science B.V.

Patterning of the vertebrate ventral spinal cord

We review investigations that have lead to a model of how the ventral spinal cord of higher vertebrate embryos is patterned during development. Central to this model is the secreted morphogen protein, Sonic hedgehog. There is now considerable evidence that this molecule acts in a concentration-dependent manner to direct the development of the spinal cord. Recent studies have suggested that two classes of homeodomain proteins are induced by threshold concentrations of Sonic hedgehog. Reciprocal inhibition between the two classes acts to convert the continuous gradient of Sonic hedgehog into defined domains of transcription factor expression. However, a number of aspects of ventral spinal cord patterning remain to be elucidated. Some issues currently under investigation involve temporal aspects of Shh-signalling, the role of other signals in ventral patterning and the characterisation of ventral interneurons. In this review, we discuss the current state of knowledge of these issues and present some preliminary studies aimed at furthering understanding of these processes in spinal cord patterning.

Distributional profiles of homologous open reading frames among bacterial phyla: implications for vertical and lateral transmission

If open reading frames (ORFs) have been transmitted primarily by vertical descent, the distributional profile of orthologues of each ORF should be congruent with the organismal tree or a subtree thereof. Distributional patterns not reconciled parsimoniously with tree-like descent and loss are prima facie evidence of lateral gene transfer. Herein, a rigorous criterion for recognizing ORF distributions is described and implemented; it does not require the inference of phylogenetic trees, nor does it assume any specific tree. Because lineage-specific differences in rates of sequence change can also generate unexpected distributional patterns, rate artefacts, were controlled for by requiring pairwise matches between ORFs to exceed a rigorous inclusion threshold, but absence of a match was assessed against a more-permissive exclusion threshold. Applying this dual-threshold criterion to cross-domain and cross-phylum distributional patterns for ORFs in 23 bacterial genomes, a relative abundance of ORFs was observed that find a match in exactly seven other bacterial phyla; 94-99% of these ORFs also find matches among the Archaea and/or Eukarya. In the larger (and some smaller) bacterial genomes, ORFs that find matches in exactly one other bacterial phylum are also relatively abundant, but fewer of these have non-bacterial homologues; most of their matches within the Bacteria are to the Proteobacteria and/or Firmicutes, which cannot be sister lineages to all bacteria. ORFs that are neither distributed universally among the Bacteria, nor necessarily shared with topologically adjacent lineages, are preferentially enriched in large bacterial genomes.

A quantitative state and transition model for the Mitchell grasslands of central western Queensland

Concerns of reduced productivity and land degradation in the Mitchell grasslands of central western Queensland were addressed through a range monitoring program to interpret condition and trend. Botanical and eclaphic parameters were recorded along piosphere and grazing gradients, and across fenceline impact areas, to maximise changes resulting from grazing. The Degradation Gradient Method was used in conjunction with State and Transition Models to develop models of rangeland dynamics and condition. States were found to be ordered along a degradation gradient, indicator species developed according to rainfall trends and transitions determined from field data and available literature. Astrebla spp. abundance declined with declining range condition and increasing grazing pressure, while annual grasses and forbs increased in dominance under poor range condition. Soil erosion increased and litter decreased with decreasing range condition. An approach to quantitatively define states within a variable rainfall environment based upon a time-series ordination analysis is described. The derived model could provide the interpretive framework necessary to integrate on-ground monitoring, remote sensing and geographic information systems to trace states and transitions at the paddock scale. However, further work is needed to determine the full catalogue of states and transitions and to refine the model for application at the paddock scale.

delta N-15 values of tropical savanna and monsoon forest species reflect root specialisations and soil nitrogen status

A large number of herbaceous and woody plants from tropical woodland, savanna, and monsoon forest were analysed to determine the impact of environmental factors (nutrient and water availability, fire) and biological factors (microbial associations, systematics) on plant delta(15)N values. Foliar delta(15)N values of herbaceous and woody species were not related to growth form or phenology, but a strong relationship existed between mycorrhizal status and plant delta(15)N. In woodland and savanna, woody species with ectomycorrhizal (ECM) associations and putative N-2-fixing species with ECM/arbuscular (AM) associations had lowest foliar delta(15)N values (1.0-0.6parts per thousand), AM species had mostly intermediate delta(15)N values (average +0.6parts per thousand), while non-mycorrhizal Proteaceae had highest delta(15)N values (+2.9 to +4.1parts per thousand). Similar differences in foliar delta(15)N were observed between AM (average 0.1 and 0.2parts per thousand) and non-mycorrhizal (average +0.8 and +0.3parts per thousand) herbaceous species in woodland and savanna. Leguminous savanna species had significantly higher leaf N contents (1.8-2.5% N) than non-fixing species (0.9-1.2% N) indicating substantial N acquisition via N-2 fixation. Monsoon forest species had similar leaf N contents (average 2.4% N) and positive delta(15)N values (+0.9 to +2.4parts per thousand). Soil nitrification and plant NO3- use was substantially higher in monsoon forest than in woodland or savanna. In the studied communities, higher soil N content and nitrification rates were associated with more positive soil delta(15)N and plant delta(15)N. In support of this notion, Ficus, a high NO3- using taxa associated with NO3- rich sites in the savanna, had the highest delta(15)N values of all AM species in the savanna. delta(15)N of xylem sap was examined as a tool for studying plant delta(15)N relations. delta(15)N of xylem sap varied seasonally and between differently aged Acacia and other savanna species. Plants from annually burnt savanna had significantly higher delta(15)N values compared to plants from less frequently burnt savanna, suggesting that foliar N-15 natural abundance could be used as marker for assessing historic fire regimes. Australian woodland and savanna species had low leaf delta(15)N and N content compared to species from equivalent African communities indicating that Australian biota are the more N depauperate. The largest differences in leaf delta(15)N occurred between the dominant ECM Australian and African savanna (miombo) species, which were depleted and enriched in N-15, respectively. While the depleted delta(15)N of Australian ECM species are similar to those of previous reports on ECM species in natural plant communities, the N-15-enriched delta(15)N of African ECM species represent an anomaly.

Depletion of benthic invertebrates by bar-tailed godwits Limosa lapponica in a subtropical estuary

This study (1) investigated functional (capture rate, foraging success) and numerical (density) responses of bar-tailed godwits Limosa lapponica to an experimental decrease in densities of their prey, and (2) estimated seasonal depletion of the stock of their main prey, the mictyrid crab Mictyris longicarpus, in a subtropical estuary. It was predicted that if intake rates of the godwits are in the vicinity of the gradient section of a functional response curve, i.e. are directly determined by prey density, they will respond rapidly to experimental reduction in the density of their prey. Bar-tailed godwits did respond rapidly, both functionally and numerically, to a decrease in the density of M longicarpus, indicating that their intake rate was limited by food availability. The estimated seasonal depletion of the stocks of Mictyris by the godwits was 88 % of the initial standing stock. Despite the virtual disappearance of Mictyris from sediment samples through the course of a non-breeding season, local densities of godwits did not change between October and March, implying that adequate rates of intake could be maintained throughout their residence period.