HIV capsid is a tractable target for small molecule therapeutic intervention

Despite a high current standard of care in antiretroviral therapy for HIV, multidrug-resistant strains continue to emerge, underscoring the need for additional novel mechanism inhibitors that will offer expanded therapeutic options in the clinic. We report a new class of small molecule antiretroviral compounds that directly target HIV-1 capsid (CA) via a novel mechanism of action. The compounds exhibit potent antiviral activity against HIV-1 laboratory strains, clinical isolates, and HIV-2, and inhibit both early and late events in the viral replication cycle. We present mechanistic studies indicating that these early and late activities result from the compound affecting viral uncoating and assembly, respectively. We show that amino acid substitutions in the N-terminal domain of HIV-1 CA are sufficient to confer resistance to this class of compounds, identifying CA as the target in infected cells. A high-resolution co-crystal structure of the compound bound to HIV-1 CA reveals a novel binding pocket in the N-terminal domain of the protein. Our data demonstrate that broad-spectrum antiviral activity can be achieved by targeting this new binding site and reveal HIV CA as a tractable drug target for HIV therapy.     

Dispersal patterns of an Australian gall-forming thrips and its host tree (Oncothrips tepperi and Acacia oswaldii )

Abstract. 1. Clark and Evans' nearest-neighbour method was used to evaluate the distribution pattern of Acacia oswaldii and Oncothrips tepperi populations.
2. A random distribution pattern was supported statistically for both the host trees and trees that were parasitised by O. tepperi .
3. It is therefore likely that dispersal distances of colonising thrips are large enough to be able to traverse the 1 km² study site.     

Use of seagrass wrack in restoring disturbed Australian saltmarshes

Summary Saltmarshes are threatened in many parts of the world, considered wastelands rather than valued as wetlands. This is particularly true in urbanized parts of Australia. Approximately 85% of saltmarsh has been lost or severely damaged in the Tuggerah Lakes of the Central Coast of New South Wales, Australia. Wrack (rotting vegetation) can promote diversity of saltmarshes, by shading soil and reducing physical stress and/or by providing nutrients to nutrient‐poor soil. In many parts of the world, wrack derives from the saltmarsh plants. On the shores of the Tuggerah Lakes, large amounts of seagrass wrack (with varying amounts of macro‐algae), are stranded along the shoreline and in the remaining saltmarshes. This is perceived to reduce public amenity and have no ecological value. Thus, the local council is under pressure to remove wrack. This experimental study examined the influence of wrack on cover and diversity of saltmarsh and provided preliminary data on changes to organic matter and benthos in the sediment. On average, there was a rapid increase in biomass of the dominant plant, Sarcocornia quinqueflora, in experimental areas where wrack was added, although the response was patchy, but there was no change in diversity over 22 months. There were limited changes to benthos, but no changes to nutrients in the sediment in wracked compared to unwracked treatments. Nevertheless, increased biomass in dominant species may reduce physical stress for other smaller saltmarsh plants. This study is part of a programme examining the interaction between the persistence and growth of saltmarshes and recycling of wrack in the lakes. The ultimate aim is to develop ecologically and economically sound plans of management for an increasingly urbanized area of the NSW coast.     

Effects of nest-box density on the behavior of Tree Swallows during nest building

ABSTRACT.   In territorial species, increased density is often linked to an increase in aggressive interactions, which may result in trade-offs between competitive behavior and nest construction. We examined the impact of nesting in areas of high-density versus low-density nest boxes on conspecific interactions and nest-building effort in a population of Tree Swallows ( Tachycineta bicolor ). We also examined whether expected differences in behavior related to variation in nest quality and reproductive success in high-density and low-density areas. No differences in either nest-building behavior or reproductive success were observed between areas of high-density and low-density boxes, but there was a tendency of more frequent behavioral interactions at high density. Similarly, there was a significant difference between pairs defending single nest boxes and those defending multiple nest boxes in the number of interactions with conspecifics. These results suggest that although there may be more competition with conspecifics for Tree Swallows nesting at high density, this does not appear to affect either nest quality or reproductive success.     

Endocrine regulation of energy metabolism by the skeleton

The regulation of bone remodeling by an adipocyte-derived hormone implies that bone may exert a feedback control of energy homeostasis. To test this hypothesis we looked for genes expressed in osteoblasts, encoding signaling molecules and affecting energy metabolism. We show here that mice lacking the protein tyrosine phosphatase OST-PTP are hypoglycemic and are protected from obesity and glucose intolerance because of an increase in beta-cell proliferation, insulin secretion, and insulin sensitivity. In contrast, mice lacking the osteoblast-secreted molecule osteocalcin display decreased beta-cell proliferation, glucose intolerance, and insulin resistance. Removing one Osteocalcin allele from OST-PTP-deficient mice corrects their metabolic phenotype. Ex vivo, osteocalcin can stimulate CyclinD1 and Insulin expression in beta-cells and Adiponectin, an insulin-sensitizing adipokine, in adipocytes; in vivo osteocalcin can improve glucose tolerance. By revealing that the skeleton exerts an endocrine regulation of sugar homeostasis this study expands the biological importance of this organ and our understanding of energy metabolism.     

Comparative analysis of virulence genes, genetic diversity, and phylogeny of commensal and enterotoxigenic Escherichia coli isolates from weaned pigs

If the acquisition of virulence genes (VGs) for pathogenicity were not solely acquired through horizontal gene transfers of pathogenicity islands, transposons, and phages, then clonal clusters of enterotoxigenic Escherichia coli (ETEC) would contain few or even none of the VGs found in strains responsible for extraintestinal infections. To evaluate this possibility, 47 postweaning diarrhea (PWD) ETEC strains from different geographical origins and 158 commensal E. coli isolates from the gastrointestinal tracts of eight group-housed healthy pigs were screened for 36 extraintestinal and 18 enteric VGs using multiplex PCR assays. Of 36 extraintestinal VGs, only 8 were detected (fimH, traT, fyuA, hlyA, kpsMtII, k5, iha, and ompT) in the ETEC collection. Among these, hlyA (alpha-hemolysin) and iha (nonhemagglutinating adhesin) occurred significantly more frequently among the ETEC isolates than in the commensal isolates. Clustering analysis based on the VG profiles separated commensal and ETEC isolates and even differentiated serogroup O141 from O149. On the other hand, pulsed-field gel electrophoresis (PFGE) successfully clustered ETEC isolates according to both serotype and geographical origin. In contrast, the commensal isolates were heterogeneous with respect to both serotype and DNA fingerprint. This study has validated the use of VG profiling to examine pathogenic relationships between porcine ETEC isolates. The clonal relationships of these isolates can be further clarified by PFGE fingerprinting. The presence of extraintestinal VGs in porcine ETEC confirmed the hypothesis that individual virulence gene acquisitions can occur concurrently against a background of horizontal gene transfers of pathogenicity islands. Over time, this could enable specific clonotypes to respond to host selection pressure and to evolve into new strains with increased virulence.     

Sensitive and specific molecular detection of Burkholderia pseudomallei, the causative agent of melioidosis, in the soil of tropical northern Australia

Burkholderia pseudomallei, the cause of the severe disease melioidosis in humans and animals, is a gram-negative saprophyte living in soil and water of areas of endemicity such as tropical northern Australia and Southeast Asia. Infection occurs mainly by contact with wet contaminated soil. The environmental distribution of B. pseudomallei in northern Australia is still unclear. We developed and evaluated a direct soil B. pseudomallei DNA detection method based on the recently published real-time PCR targeting the B. pseudomallei type III secretion system. The method was evaluated by inoculating different soil types with B. pseudomallei dilution series and by comparing B. pseudomallei detection rate with culture-based detection rate for 104 randomly collected soil samples from the Darwin rural area in northern Australia. We found that direct soil B. pseudomallei DNA detection not only was substantially faster than culture but also proved to be more sensitive with no evident false-positive results. This assay provides a new tool to detect B. pseudomallei in soil samples in a fast and highly sensitive and specific manner and is applicable for large-scale B. pseudomallei environmental screening studies or in outbreak situations. Furthermore, analysis of the 104 collected soil samples revealed a significant association between B. pseudomallei-positive sites and the presence of animals at these locations and also with moist, reddish brown-to-reddish gray soils.     

A Biochemical Guide to Yeast Adhesins: Glycoproteins for Social and Antisocial Occasions

Fungi are nonmotile eukaryotes that rely on their adhesins for selective interaction with the environment and with other fungal cells. Glycosylphosphatidylinositol (GPI)-cross-linked adhesins have essential roles in mating, colony morphology, host-pathogen interactions, and biofilm formation. We review the structure and binding properties of cell wall-bound adhesins of ascomycetous yeasts and relate them to their effects on cellular interactions, with particular emphasis on the agglutinins and flocculins of Saccharomyces and the Als proteins of Candida. These glycoproteins share common structural motifs tailored to surface activity and biological function. After being secreted to the outer face of the plasma membrane, they are covalently anchored in the wall through modified GPI anchors, with their binding domains elevated beyond the wall surface on highly glycosylated extended stalks. N-terminal globular domains bind peptide or sugar ligands, with between millimolar and nanomolar affinities. These affinities and the high density of adhesins and ligands at the cell surface determine microscopic and macroscopic characteristics of cell-cell associations. Central domains often include Thr-rich tandemly repeated sequences that are highly glycosylated. These domains potentiate cell-to-cell binding, but the molecular mechanism of such an association is not yet clear. These repeats also mediate recombination between repeats and between genes. The high levels of recombination and epigenetic regulation are sources of variation which enable the population to continually exploit new niches and resources.