Differential regulation of twitching motility and elastase production by Vfr in Pseudomonas aeruginosa

Vfr, a homolog of Escherichia coli cyclic AMP (cAMP) receptor protein, has been shown to regulate quorum sensing, exotoxin A production, and regA transcription in Pseudomonas aeruginosa. We identified a twitching motility-defective mutant that carries a transposon insertion in vfr and confirmed that vfr is required for twitching motility by construction of an independent allelic deletion-replacement mutant of vfr that exhibited the same phenotype, as well as by the restoration of normal twitching motility by complementation of these mutants with wild-type vfr. Vfr-null mutants exhibited severely reduced twitching motility with barely detectable levels of type IV pili, as well as loss of elastase production and altered pyocyanin production. We also identified reduced-twitching variants of quorum-sensing mutants (PAK lasl::Tc) with a spontaneous deletion in vfr (S. A. Beatson, C. B. Whitchurch, A. B. T. Semmler, and J. S. Mattick, J. Bacteriol., 184:3598-3604,2002), the net result of which was the loss of five residues (EQERS) from the putative cAMP-binding pocket or Vfr. This allele (VfrDeltaEQERS) was capable of restoring elastase and pyocyanin production to wild-type levels in vfr-null mutants but not their defects in twitching motility. Furthermore, structural analysis of Vfr and VfrDeltaEQERS in relation to E. coli CRP suggests that Vfr is capable of binding both cAMP and cyclic GMP whereas VfrDeltaEQERS is only capable of responding to cAMP. We suggest that Vfr controls twitching motility and quorum sensing via independent pathways in response to these different signals, bound by the same cyclic nucleotide monophosphate-binding pocket.

Genotypic variation in proanthocyanidin status in the Leucaena genus

The proanthocyanidin (PA) status of 116 accessions from the Leucaena genus representing 21 species, 6 subspecies, 3 varieties and 4 interspecific hybrids was evaluated under uniform environmental and experimental conditions at Redland Bay, Queensland, Australia in October 1997. The PA content of lyophilized youngest fully expanded leaves was measured spectrophotometrically by the butanol/HCl assay referenced to L. leucocephala ssp. glabrata standard PA and expressed as L. leucocephala ssp. glabrata PA equivalents (LLPAE). Considerable interspecific variation in PA concentration existed within the genus, ranging from 0-339 g LLPAE/kg dry matter (DM). Taxa including L. confertiflora, L. cuspidata, L. esculenta and L. greggii contained very high (> 180 g LLPAE/kg DM) PA concentrations. Similarly, many agronomically superior accessions from L. diversifolia, L. pallida and L. trichandra contained extremely high (up to 250 g LLPAE/kg DM) PA concentrations, although these taxa exhibited wide intraspecific variation in PA content offering the potential to select accessions with lower (120-160 g LLPAE/kg DM) PA content. Commercial cultivars of L. leucocephala ssp. glabrata, known to produce forage of superior quality, contained low amounts of PA (33-39 g LLPAE/kg DM). Artificial interspecific hybrids had PA contents intermediate to those of both parents, Lesser-known taxa. including L. collinsii, L. lanceolata, L. lempirana, L. macrophylla, L. magnifica, L. multicapitula, L. salvadorensis and L. trichodes, contained undetectable to low (0-36 g LLPAE/kg DM) quantities of PA and have potential as parents to breed interspecific hybrids of low PA status and superior forage quality. Extractable PA was the dominant PA component, accounting for 91% of total PA within the genus. Regression analysis of accession ranks from different experiments compared to these results indicated that genetic regulation of Leucaena spp. PA content was consistent (P < 0.01) under different edapho-climatic environments. The distribution of PA within the Leucaena genus did not concur with the predictions of various evolutionary and phylogenetic plant defence theories.

Associations between human liver and kidney cadmium content and immunochemically detected CYP4A11 apoprotein

This present study was undertaken to assess potential effects of cadmium on CYP4A11 apoprotein in human liver and kidney as detected by Western blotting using a highly specific anti-peptide antibody. Liver and kidney cortex samples were autopsy specimens of 37 individuals (26 mates and I I females) whose ages ranged from 3 to 89 years. All were Caucasians who had not been exposed to cadmium in the workplace. Reduced CYP4A11 apoprotein levels were found in chronic hepatitis samples and in liver samples showing fatty changes. In contrast, increased CYP4A11 apoprotein levels were found in liver samples having higher cadmium content compared to the lower cadmium content samples. Increased CYP4A11 levels were also found in liver samples from female donors, compared to male donors; the difference being attributable to higher female liver cadmium burden. In distinction to liver, lowered CYP4A11 levels were seen in the kidney cortex samples which have high cadmium content, It is proposed here that the difference between the absolute cadmium burden of the liver and kidney samples may be responsible for the different patterns of expression of CYP4A11 in these two tissues. Further, since cadmium exposure may be associated with derangement in blood pressure control, it is interesting to note the possible relationship between altered CYP4A11-dependent production of arachidonic acid hydroxy and epoxy metabolites in kidney cortex and altered control of blood pressure. Our findings provide a possible link between these observations. (C) 2002 Elsevier Science Inc. All rights reserved.

Future contributions of crop modelling - from heuristics and supporting decision making to understanding genetic regulation and aiding crop improvement

Crop modelling has evolved over the last 30 or so years in concert with advances in crop physiology, crop ecology and computing technology. Having reached a respectable degree of acceptance, it is appropriate to review briefly the course of developments in crop modelling and to project what might be major contributions of crop modelling in the future. Two major opportunities are envisioned for increased modelling activity in the future. One opportunity is in a continuing central, heuristic role to support scientific investigation, to facilitate decision making by crop managers, and to aid in education. Heuristic activities will also extend to the broader system-level issues of environmental and ecological aspects of crop production. The second opportunity is projected as a prime contributor in understanding and advancing the genetic regulation of plant performance and plant improvement. Physiological dissection and modelling of traits provides an avenue by which crop modelling could contribute to enhancing integration of molecular genetic technologies in crop improvement. Crown Copyright (C) 2002 Published by Elsevier Science B.V. All rights reserved.

Selenium content of Australian foods: A review of literature values

The aim of this study was to review the published literature values for the selenium content of Australian foods. A secondary aim was to compare the results for Australian foods with food composition data from international sources to investigate the extent of geographical variation. Published food composition data sources for the selenium content in Australian foods were identified and assessed for data quality using established criteria. The selenium content is available for 148 individual food items. The highest values found are for fish (12.0-63.2 mug/100 g), meats (4.75-37.9 mug/100 g) and eggs (9.00-41.4 mug/100 g), followed by cereals (1.00-20.3 mug/100 g). Moderate levels are seen in dairy products (2.00-7.89 mug/100 g) while most fruits and vegetables have low levels (trace-3.27 mug/100 g). High selenium foods show the greatest level of geographical variation, with foods from the United States generally having higher selenium levels than Australian foods and foods from the United Kingdom and New Zealand having lower levels. This is the first attempt to review the available literature for selenium composition of Australian foods. These data serve as an interim measure for the assessment of selenium intake for use in epidemiological studies of diet-disease relationships. (C) 2002 Published by Elsevier Science Ltd.

Preparing students for the virtual organisation: an evaluation of learning with virtual learning technologies

The paper explores the development of learning behaviours in a virtual management course and the factors that impacted on this development. Data suggest that most teams experienced three kinds of learning behaviours – social, operational and content learning. We propose that the need for technical expertise and team participation will vary during these different stages of learning. Addressing the characteristics of these stages, we comment on the development of a ‘completion phase’ of team development. We argue that the extent to which teams demonstrate different learning stages has a significant impact on the development of on-line learning behaviours. Discussing these results, we suggest why different teams develop distinct learning behaviours, with accordant emphasis on teaching as a moderating and co ordinating role, despite current virtual team pedagogical expectations.

Regulation of the mouse Nas1 promoter by vitamin D and thyroid hormone

The renal sodium-sulfate cotransporter, NaSi-1, a protein implicated to control serum sulfate levels, has been shown to be regulated in vivo by 1,25-dihydroxyvitamin D-3 (1,25-(OH)(2)D-3) and tri-iodothyronine (T-3). Recently, we cloned the mouse NaSi-1 gene (Nas1) and in the present study identified a 1,25-(OH)(2)D-3- and T-3-responsive element located within the Nas1 promoter. Mutational analysis of the Nas1 promoter resulted in identification of a direct repeat 6-type vitamin-D-responsive element (DR6 VDRE) at -525 to -508 and an imperfect inverted repeat 0-type T-3-responsive element (IR0 T3RE) at -436 to -425 which conferred 1,25(OH)(2)D-3 and T3 responsiveness, respectively. In summary, we have identified responsive elements that mediate the enhanced transcription of Nas1 by 1,25-(OH)(2)D-3 and T-3, and these mechanisms may provide important clues to the physiological control of sulfate homeostasis.

Transcriptional regulation of the sodium-sulfate cotransporter NaSi-1 gene

Inorganic sulfate is one of the most abundant anions in mammalian plasma and is essential for proper cell growth and development, as well as detoxification and activation of many biological compounds. To date, little is understood how physiological levels of sulfate are maintained in the body. Our studies, and of others, have identified the NAS(i)-1 protein to be a functional sulfate transporter in the kidney and intestine, and due to this localization, constitutes a strong candidate gene for maintaining body sulfate homeostasis. Several factors, including hormones and metabolic conditions, have been shown to alter NAS(i)-1 mRNA and protein levels in vivo. In this study, we describe the transcriptional regulation of NaSi-1, with a focus on the mouse NaSi-1 gene (Nas1) that was recently cloned in our laboratory. Vitamin D (1,25-(OH)(2)D-3) and thyroid hormone (T-3) led to an increase in Nas1 promoter activity in OK cells. Mutational analysis of the Nas1 promoter resulted in identification of a direct repeat 6-type vitamin-D-responsive element (DR6 VDRE) at -525 to -508 and an imperfect inverted repeat 0-type T-3 responsive element (IRO T3RE) at -426 to -425 which conferred 1,25-(OH)(2)D-3 and T-3 responsiveness respectively. These findings suggest for vitamin D and thyroid hormone regulation of NaSi-1, may provide important clues to the physiological control of sulfate homeostasis.

Application of quantitative analytical electron microscopy to the mineral content of insect cuticle

Quantification of calcium in the cuticle of the fly larva Exeretonevra angustifrons was undertaken at the micron scale using wavelength dispersive X-ray microanalysis, analytical standards, and a full matrix correction. Calcium and phosphorus were found to be present in the exoskeleton in a ratio that indicates amorphous calcium phosphate. This was confirmed through electron diffraction of the calcium-containing tissue. Due to the pragmatic difficulties of measuring light elements, it is not uncommon in the field of entomology to neglect the use of matrix corrections when performing microanalysis of bulk insect specimens. To determine, firstly, whether such a strategy affects the outcome and secondly, which matrix correction is preferable, phi-rho (z) and ZAF matrix corrections were contrasted with each other and without matrix correction. The best estimate of the mineral phase was found to be given by using the phi-rho (z) correction. When no correction was made, the ratio of Ca to P fell outside the range for amorphous calcium phosphate, possibly leading to flawed interpretation of the mineral form when used on its own.

Indirect evidence of density-dependent population regulation in Aponomma hydrosauri (Acari : Ixodidae), an ectoparasite of reptiles

The extent to which density-dependent processes regulate natural populations is the subject of an ongoing debate. We contribute evidence to this debate showing that density-dependent processes influence the population dynamics of the ectoparasite Aponomma hydrosauri (Acari: Ixodidae), a tick species that infests reptiles in Australia. The first piece of evidence comes from an unusually long-term dataset on the distribution of ticks among individual hosts. If density-dependent processes are influencing either host mortality or vital rates of the parasite population, and those distributions can be approximated with negative binomial distributions, then general host-parasite models predict that the aggregation coefficient of the parasite distribution will increase with the average intensity of infections. We fit negative binomial distributions to the frequency distributions of ticks on hosts, and find that the estimated aggregation coefficient k increases with increasing average tick density. This pattern indirectly implies that one or more vital rates of the tick population must be changing with increasing tick density, because mortality rates of the tick's main host, the sleepy lizard, Tiliqua rugosa, are unaffected by changes in tick burdens. Our second piece of evidence is a re-analysis of experimental data on the attachment success of individual ticks to lizard hosts using generalized linear modelling. The probability of successful engorgement decreases with increasing numbers of ticks attached to a host. This is direct evidence of a density-dependent process that could lead to an increase in the aggregation coefficient of tick distributions described earlier. The population-scale increase in the aggregation coefficient is indirect evidence of a density-dependent process or processes sufficiently strong to produce a population-wide pattern, and thus also likely to influence population regulation. The direct observation of a density-dependent process is evidence of at least part of the responsible mechanism.

Variables affecting the fine grinding of minerals using stirred mills

Base metal resources are becoming more fine-grained and refractory and minerals separation processes require these ores to be milled to increasingly finer sizes. To cope with very fine grinding to below a P-80 of approximately 15 mum stirred milling technology has been adopted from other industries Neither this technology, nor the basic concepts of fine grinding, are well understood by the minerals processing industry. Laboratory studies were therefore carried out in order to investigate fine milling using different types of stirred mills. The variables analysed were stirrer speed, grinding media type and size, slurry solids content as well as the feed and product size. The results of the testwork have shown that all of these variables affect the grinding efficiency. The ratio of media size to material size was found to be of particular significance. The results were also analysed using the stress intensity approach and the optimum stress intensity ranges for the most efficient grinding were determined. Application of the results for process optimisation in the industrial size units is also discussed in this paper. (C) 2003 Elsevier Science Ltd. All rights reserved.