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AbstractSeveral different species of freshwater Bryozoa, belonging to the genera Plumatella, Rumarcanellaand Fredericella, were detected within the Northern Mallee Pipeline (NMP) system in Victoria, Australia, that required definitive identification. These organisms produce asexual buds called statoblasts, with valves composed of sclerotised chitin that bear minute micro-ornamentations of considerable taxonomical significance. Imaging and analysis of these distinctive micro-ornamentations using scanning electron microscopy (SEM) is often employed for species identification. Meticulous preparation of statoblast samples is therefore required that necessitates the removal of adhering debris, dehydration and drying—whilst mitigating specimen damage and distortion. This technical note describes an approach whereby each of these three steps have been individually designed to be as benign as possible, using mild detergent/sonication to remove debris, a gradual and gentle dehydration procedure using ethanol, and critical point drying. For the overall process, these methods are chosen to optimise control and to minimise the use of harsh and hazardous chemicals.

AbstractFive species of freshwater bryozoans were identified in the Northern Mallee Pipeline (NMP) system in Victoria, Australia, from statoblast morphology using scanning electron microscopy. The five species were Plumatella emarginata, Fredericella australiensis, Plumatella reticulata, Rumarcanella minutaand Fredericella sultana. The latter three of these have never previously been reported as occurring on the Australian continent and samples have been preserved for safekeeping at Victoria University. The statoblasts were isolated from samples collected as intact colonies from filters and membranes at a number of different locations.

AbstractThe last 12 years has seen extreme drought in western Victoria. This has impacted on the area in many ways, but none more so than in the provision of basic water supplies to people. To meet the challenge of drought, headworks storages have had to be operated at record low levels, severe water restrictions imposed, water carting programs established, alternative sources of water, and new technologies developed and used. Significant changes have also been made to the water supply infrastructure in the region, most notably the Northern-Mallee and Wimmera-Mallee Pipelines. This paper relates the story of how water resources were managed and bulk water was delivered to around 70,000 customers over a geographic spread of 62,000 km2, or about 30% of Victoria. Discussion on the social, environmental and economic impacts on the region are also provided.

Tissue repair begins with the formation of a blood clot, and moves through the stages of chemotaxis, migration of neutrophils, macrophages and fibroblasts, angio genesis, formation of a provisional matrix, synthesis of collagen, assembly of collagen matrix, migration and proliferation of keratinocytes and, finally, wound closure. Each process requires groups of cells to perform various functions in consort and is controlled by growth factors and cytokines through cell signalling.

This study investigated relationships between birds and characteristics of woodland remnants in the central wheat/sheep belt of New South Wales, in order to provide information for conservation management of woodland birds and their habitats in this region. Birds were surveyed in standard area plots in 36 woodland remnants, which ranged from 1 to 1 376 hectares in size, in the central Lachlan catchment in the wheat/sheep belt of New South Wales. Habitat (e.g., shrub cover) and landscape (measures of isolation) characteristics of the remnants were recorded also. Presence/absence occurrences of 20 or more bird species, mostly woodland birds, were positively related to remnant area, shrub cover, number of shrub species, pine cover, fallen logs and branches, and number of remnants within two and five kilometres of the survey remnant. Species of birds that are habitat generalists showed negative or no relationships with these habitat and landscape variables. Richness of all bird species was positively related to remnant area and shrub cover. Number of woodland bird species was positively related to remnant area, shrub cover and tree hollows. Lower species diversities in smaller remnants were most likely caused by lower chances of survival or of successful breeding in small remnants, effects of isolation on the ability of birds to recolonize smaller remnants, small remnants being of insufficient area for some species to form territories, and generally poor habitat quality in small remnants. Retention, management and enhancement of woodland remnants 10 to 20 hectares or larger in size with good understorey cover are necessary for the future viability of woodland birds in the wheat/sheep belt of New South Wales.

Histone methylation is known to be associated with both transcriptionally active and repressive chromatin states. Recent studies have identified SET domain-containing proteins such as SUV39H1 and Clr4 as mediators of H3 lysine 9 (Lys9) methylation and heterochromatin formation. Interestingly, H3 Lys9 methylation is not observed from bulk histones isolated from asynchronous populations of Saccharomyces cerevisiae or Tetrahymena thermophila. In contrast, H3 lysine 4 (Lys4) methylation is a predominant modification in these smaller eukaryotes. To identify the responsible methyltransferase(s) and to gain insight into the function of H3 Lys4 methylation, we have developed a histone H3 Lys4 methyl-specific antiserum. With this antiserum, we show that deletion of SET1, but not of other putative SET domain-containing genes, in S. cerevisiae, results in the complete abolishment of H3 Lys4 methylation in vivo. Furthermore, loss of H3 Lys4 methylation in a set1 Delta strain can be rescued by SET1. Analysis of histone H3 mutations at Lys4 revealed a slow-growth defect similar to a set1 Delta strain. Chromatin immunoprecipitation assays show that H3 Lys4 methylation is present at the rDNA locus and that Set1-mediated H3 Lys4 methylation is required for repression of RNA polymerase II transcription within rDNA. Taken together, these data suggest that Set1-mediated H3 Lys4 methylation is required for normal cell growth and transcriptional silencing.

Human immunodeficiency virus Nef is a small myristylated protein that plays a critical role in AIDS progression. Nef binds with high affinity to the SH3 domain of the myeloid-restricted tyrosine kinase Hck in vitro, identifying this Src-related kinase as a possible cellular target for Nef in macrophages. Here we show that Nef activates endogenous Hck in the granulocyte-macrophage colony-stimulating factor-dependent myeloid cell line, TF-1. Unexpectedly, Nef induced cytokine-independent TF-1 cell outgrowth and constitutive activation of the Stat3 transcription factor. Induction of survival required the Nef SH3 binding and membrane-targeting motifs and was blocked by dominant-negative Stat3 mutants. Nef also stimulated Stat3 activation in primary human macrophages, providing evidence for Stat3 as a Nef effector in a target cell for human immunodeficiency virus.

IT is the year 2020. Farmers have broken the record for the number of threatened species recorded on private property in a single year. Five species of woodland bird are removed from threatened species lists. The area of private land managed for biodiversity now exceeds the area of public land managed for biodiversity. Farm income from biodiversity is greater than wheat. The Biodiversity Growers Association calls for the environment levy on Australian taxpayers to be increased. Drought relief payments are at an all time low. Retiring head of government conservation agency described as "visionary" by farmers' representatives.

The N-terminal fragment 1-34 of parathyroid hormone (PTH), administered intermittently, results in increased bone formation in patients with osteoporosis. PTH and a related molecule, parathyroid hormone-related peptide (PTHrP), act on cells via a common PTH/PTHrP receptor. To define more precisely the ligand-receptor interactions, we have crystallized human PTH (hPTH)-(1-34) and determined the structure to 0.9-A resolution. hPTH-(1-34) crystallizes as a slightly bent, long helical dimer. Analysis reveals that the extended helical conformation of hPTH-(1-34) is the likely bioactive conformation. We have developed molecular models for the interaction of hPTH-(1-34) and hPTHrP-(1-34) with the PTH/PTHrP receptor. A receptor binding pocket for the N terminus of hPTH-(1-34) and a hydrophobic interface with the receptor for the C terminus of hPTH-(1-34) are proposed.

To enhance the potency of 1,2-dibenzamidobenzene-derived inhibitors of factor Xa (fXa), an amidine substituent was incorporated on one of the benzoyl side chains to interact with Asp189 in the S1 specificity pocket. Lead molecule 1 was docked into the active site of fXa to facilitate inhibitor design. Subsequently, iterative SAR studies and molecular modeling led to a 1000-fold increase in fXa affinity and a refined model of the new inhibitors in the fXa active site. Strong support for the computational model was achieved through the acquisition of an X-ray crystal structure using thrombin as a surrogate protein. The amidines in this series show high levels of selectivity for the inhibition of fXa relative to other trypsin-like serine proteases. Furthermore, the fXa affinity of compounds in this series (Kass = 50−500 × 10⁶ L/mol) translates effectively into both anticoagulant activity in vitro and antithrombotic activity in vivo.

Biochemical and structural studies of Src and related kinases demonstrate that two intramolecular interactions suppress kinase activity. These interactions involve binding of the SH2 domain to a phosphotyrosine residue in the C-terminal tail and association of the SH3 domain with a polyproline type II helix formed by amino acids linking the SH2 and kinase domains. Recent studies have shown that high affinity interaction of the SH3 domain of Hck with the human immunodeficiency virus type I Nef protein activates Hck tyrosine kinase and biological activities, suggesting a mechanism that involves disruption of the SH3-linker interaction. To test the role of this interaction in the regulation of Hck kinase activity in living cells, we substituted alanines for prolines 225 and 228 in the linker region and observed that the resulting mutant (Hck-2PA) demonstrated strong transforming activity in a Rat-2 fibroblast focus-forming assay. Hck-2PA also exhibited elevated tyrosine kinase activity in terms of autophosphorylation, endogenous substrate phosphorylation, and in an in vitro kinase assay. The transforming and kinase activities of Hck-2PA were remarkably similar to those observed with a Hck mutant activated by Phe substitution of the conserved tail Tyr residue and with wild-type Hck following co-expression with human immunodeficiency virus Nef. Introduction of the 2PA and tail mutations into a single Hck expression construct did not increase kinase or transforming activity relative to the individual mutations. These data provide new evidence that SH3-linker interaction may represent the dominant mechanism controlling Hck tyrosine kinase activity in vivo.

Tyrosine kinases of the Src family are regulated via their Src homology 2 (SH2) and SH3 domains. The Nef protein of human immunodeficiency virus-1 (HIV-1) has previously been shown to bind with high affinity and specificity in vitro to the SH3 domain of Hck, a Src family member expressed primarily in myeloid cells. However, the effect of Nef on Hck activity in living cells is unknown. Here we show that Rat-2 fibroblasts co-expressing Hck and Nef rapidly developed transformed foci, whereas control cells expressing either protein alone did not. Nef formed a stable complex with Hck and stimulated its tyrosine kinase activity in vivo. Mutagenesis of the Nef proline-rich motif essential for SH3 binding completely blocked complex formation, kinase activation, and transformation, indicating that the Nef SH3-binding function is required for its effects on Hck. These results provide direct evidence that SH3 engagement is sufficient to activate a Src family kinase in vivo and suggest that Hck may be activated by Nef in HIV-infected macrophages.

1. In hepatocytes isolated from 24 h-starved rats, urea production from ammonia was stimulated by addition of lactate, in both the presence and the absence of ornithine. The relationship of lactate concentration to the rate of urea synthesis was hyperbolic. 2. Other glucose precursors also stimulated urea production to varying degrees, but none more than lactate. Added oleate and butyrate did not stimulate urea synthesis. 3. Citrulline accumulation was largely dependent on ornithine concentration. As ornithine was increased from 0 to 40 mM, the rate of citrulline accumulation increased hyperbolically, and was half-maximal when ornithine was 8-12 mM. 4. The rate of citrulline accumulation was independent of the presence of lactate, but with pyruvate the rate increased. 5. The rate of urea production continued to increase as ornithine was varied from 0 to 40 mM. 6. It was concluded that intermediates provided by both ornithine and lactate are limiting for urea production from ammonia in isolated liver cells. It was suggested that the stimulatory effect of lactate lies in increased availability of cytosolic aspartate for condensation with citrulline.

We have devised, written and tested an implementation of the Gaussian Maximum Likelihood classification method for a commercial image processor. This has resulted in significant savings in execution time for the classification of multispectural remotely-sensed imagery, at very little cost to the accuracy, when compared to a software version of the same algorithm.

High-resolution data from the HRV (High Resolution Visible) sensors onboard the SPOT-1 satellite have been utilized for mapping semi-natural and agricultural land cover using automated digital image classification algorithms. Two methods for improving classification performance are discussed. The first technique involves the use of digital terrain information to reduce the effects of topography on spectral information while the second technique involves the classification of land-cover types using training data derived from spectral feature space. Test areas in Snowdonia and the Somerset Levels were used to evaluate the methodology and promising results were achieved. However, the low classification accuracies obtained suggest that spectral classification alone is not a suitable tool to use in the mapping of semi-natural cover types.