Cataloging altered gene expression in young and senescent cells using enhanced differential display.

Recently, a novel PCR-based technique, differential display (DD), has facilitated the study of differentially expressed genes at the mRNA level. We report here an improved version of DD, which we call Enhanced Differential Display (EDD). We have modified the technique to enhance reproducibility and to facilitate sequencing and cloning. Using EDD, we have generated and verified a catalog of genes that are differentially expressed between young and senescent human diploid fibroblasts (HDF). From 168 genetags that were identified initially, 84 could be sequenced directly from PCR amplified bands. These sequences represent 27 known genes and 37 novel genes. By Northern blot analysis we have confirmed the differential expression of a total of 23 genes (12 known, 11 novel), while 19 (seven known, 12 novel) did not show differential expression. Several of the known genes were previously observed by others to be differentially expressed between young and senescent fibroblasts, thereby validating the technique.     

Exploring stream communities in a tropical biodiversity hotspot: biodiversity,regional occupancy,niche characteristics and environmental correlates

Exploring and describing biodiversity and the mechanisms structuring it is fundamental to advancing ecology. This is particularly pertinent in understudied biogeographical regions, such as the Afrotropics, that are characterised by strong seasonal climatic shifts. We investigated the characteristics of stream biodiversity in the Niger Delta region of Nigeria, a tropical biodiversity hotspot, by examining patterns in 20 stream invertebrate communities across both the wet and dry seasons. For this, we took a multi-faceted approach accounting for the three levels of biodiversity (α, β and γ), including partitioning the nestedness and turnover components of β diversity, regional occupancy-abundance patterns, niche characteristics, and the environmental drivers of community structure. α diversity was low in these streams, with strong turnover between sites leading to high β diversity contributing to regional biodiversity, but there was little variation in communities between seasons. The proportion of sites occupied by taxa declined with increasing niche position, and decreasing niche breadth. Occupancy was predicted well by a combination of these two factors (niche position and breadth), but not mean local abundance, as the abundance-occupancy link was an upper-limit unimodal relationship. On average, community structure was linked more strongly to environmental variables in the wet season. Our findings demonstrate the clear role of spatial, but not temporal, turnover in assemblages, which likely reflects the environmental heterogeneity of this region. This is further supported by the fact that regional occupancy was mostly related to niche characteristics, particularly niche position. We emphasise the importance of continued basic and applied ecological work in this important biogeographic region to enable better protection of its biodiversity.     

Wildlife detector dogs and camera traps: a comparison of techniques for detecting feral cats

A major challenge in controlling overabundant wildlife is monitoring their populations, particularly as they decline to very low density. Camera traps and wildlife detector dogs are increasingly being used for this purpose. We compared the cost-effectiveness of these two approaches for detecting feral cats (Felis catus) on two pastoral properties in Hawke's Bay, North Island, New Zealand. One property was subject to intensive pest removal, while the other had no recent history of pest control. Camera traps and wildlife detector dogs detected cats at similar rates at both sites. The operating costs of each method were also comparable. We identify a number of advantages and disadvantages of each technique, and suggest priorities for further research.     

Quirks of dye nomenclature. 8. Methylene blue,azure and violet

Methylene blue was synthesized in 1877 and soon found application in medicine, staining for microscopy and as an industrial dye and pigment. An enormous literature has accumulated since its introduction. Early on, it was known that methylene blue could be degraded easily by demethylation; consequently, the purity of commercial samples often was low. Therefore, demethylation products, such as azures and methylene violet, also are considered here. The names and identity of the components, their varying modes of manufacture, analytical methods and their contribution to biological staining are discussed.     

Signatures of polygenic adaptation associated with climate across the range of a threatened fish species with high genetic connectivity

Adaptive differences across species’ ranges can have important implications for population persistence and conservation management decisions. Despite advances in genomic technologies, detecting adaptive variation in natural populations remains challenging. Key challenges in gene–environment association studies involve distinguishing the effects of drift from those of selection and identifying subtle signatures of polygenic adaptation. We used paired‐end restriction site‐associated DNA sequencing data (6,605 biallelic single nucleotide polymorphisms; SNPs) to examine population structure and test for signatures of adaptation across the geographic range of an iconic Australian endemic freshwater fish species, the Murray cod Maccullochella peelii. Two univariate gene–association methods identified 61 genomic regions associated with climate variation. We also tested for subtle signatures of polygenic adaptation using a multivariate method (redundancy analysis; RDA). The RDA analysis suggested that climate (temperature‐ and precipitation‐related variables) and geography had similar magnitudes of effect in shaping the distribution of SNP genotypes across the sampled range of Murray cod. Although there was poor agreement among the candidate SNPs identified by the univariate methods, the top 5% of SNPs contributing to significant RDA axes included 67% of the SNPs identified by univariate methods. We discuss the potential implications of our findings for the management of Murray cod and other species generally, particularly in relation to informing conservation actions such as translocations to improve evolutionary resilience of natural populations. Our results highlight the value of using a combination of different approaches, including polygenic methods, when testing for signatures of adaptation in landscape genomic studies.     

Effects of flooding on recruitment and dispersal of the Southern Pygmy Perch (Nannoperca australis) at a Murray River floodplain wetland

Summary With limited evidence linking Australia's Murray‐Darling Basin fish species and flooding, this study assessed annual variation in abundance and recruitment levels of a small‐bodied, threatened floodplain species, the Southern Pygmy Perch (Nannoperca australis), in floodplain habitats (creeks, lakes and wetlands) in the Barmah‐Millewa Forest, Murray River, Australia. Spring and summer sampling over a 5‐year period encompassed large hydrological variation, including 1 year of extended floodplain inundation which was largely driven by an environmental water release, and 2 years of severe regional drought. Recruitment and dispersal of Southern Pygmy Perch significantly increased during the floodplain inundation event compared with the other examined years. This study provides valuable support for an environmental water allocation benefiting a native species, and explores the link between flooding and its advantages to native fish. This suggests that the reduced flooding frequency and magnitude as a result of river regulation may well be a major contributing factor in the species’ decline in the Murray‐Darling Basin.     

Bacterial networking

A report of the ESF-EMBO Symposium Bacterial Networks (BacNet08), Sant Feliu de Guixols, Spain, 13-18 September 2008.     

Complement activation by phospholipids: the interplay of factor H and C1q

Complement proteins in blood recognize charged particles. The anionic phospholipid (aPL) cardiolipin binds both complement proteins C1q and factor H. C1q is an activator of the complement classical pathway, while factor H is an inhibitor of the alternative pathway. To examine opposing effects of C1q and factor H on complement activation by aPL, we surveyed C1q and factor H binding, and complement activation by aPL, either coated on microtitre plates or in liposomes. Both C1q and factor H bound to all aPL tested, and competed directly with each other for binding. All the aPL activated the complement classical pathway, but negligibly the alternative pathway, consistent with accepted roles of C1q and factor H. However, in this system, factor H, by competing directly with C1q for binding to aPL, acts as a direct regulator of the complement classical pathway. This regulatory mechanism is distinct from its action on the alternative pathway. Regulation of classical pathway activation by factor H was confirmed by measuring C4 activation by aPL in human sera in which the C1q:factor H molar ratio was adjusted over a wide range. Thus factor H, which is regarded as a down-regulator only of the alternative pathway, has a distinct role in downregulating activation of the classical complement pathway by aPL. A factor H homologue, β2-glycoprotein-1, also strongly inhibits C1q binding to cardiolipin. Recombinant globular domains of C1q A, B and C chains bound aPL similarly to native C1q, confirming that C1q binds aPL via its globular heads.