Relationships between native small mammals and native and introduced large herbivores

Australia has a range of native and introduced large herbivores that could affect the abundance of small mammals through direct and indirect effects. Here we study the relationship between occurrence of the introduced rusa deer (Rusa timorensis) and the native swamp wallaby (Wallabia bicolor), and the abundance of four species of native small mammals in coastal heath vegetation with varying fire history. The abundance of two species, the brown antechinus (Antechinus stuartii) and bush rat (Rattus fuscipes), was related to occurrence of large herbivores and was dependent also on fire history. Abundance of swamp rats (R. lutreolus) and New Holland mice (Pseudomys novaehollandiae) was not related to the occurrence of any of the large herbivores, and did not depend on fire history. At sites burned within the last 9 years, captures of brown antechinus were negatively related to both deer and wallaby occurrence, and captures of bush rats were negatively related to deer occurrence. However, at sites that burned more than 15 years ago, captures of brown antechinus and bush rats were not related to large herbivore occurrence. Overall there was either no relationship, or a negative one, between small mammals and the large herbivores. This mensurative study has demonstrated relationships between deer and wallabies and small mammals, with fire as an additional important factor. From the results of the current study we put forward a series of hypotheses that need to be tested by future experiments.     

The TRiC/CCT Chaperone Is Implicated in Alzheimer's Disease Based on Patient GWAS and an RNAi Screen in Aβ-Expressing Caenorhabditis elegans

The human Aβ peptide causes progressive paralysis when expressed in the muscles of the nematode worm, C. elegans. We have exploited this model of Aβ toxicity by carrying out an RNAi screen to identify genes whose reduced expression modifies the severity of this locomotor phenotype. Our initial finding was that none of the human orthologues of these worm genes is identical with the genome-wide significant GWAS genes reported to date (the “white zone”); moreover there was no identity between worm screen hits and the longer list of GWAS genes which included those with borderline levels of significance (the “grey zone”). This indicates that Aβ toxicity should not be considered as equivalent to sporadic AD. To increase the sensitivity of our analysis, we then considered the physical interactors (+1 interactome) of the products of the genes in both the worm and the white+grey zone lists. When we consider these worm and GWAS gene lists we find that 4 of the 60 worm genes have a +1 interactome overlap that is larger than expected by chance. Two of these genes form a chaperonin complex, the third is closely associated with this complex and the fourth gene codes for actin, the major substrate of the same chaperonin.     

Microengineered surface topography facilitates cell grafting from a prototype hydrogel wound dressing with antibacterial capability

Skin wounds derive therapeutic benefit from redeployment of dermal tissues, whether as split-thickness allo- and autografts or as biological dressings comprising cultured cells. However, the clinical outcome is strongly influenced by the techniques used for cell/tissue grafting and also the microbiological status of the wound. Here we report that microtopography incorporated into the surface of a novel polymeric material, derivatized with fibronectin to promote attachment and encourage motility, improved the efficiency of cell transfer onto de-epithelialized human skin ex vivo. The microtopography had two functions, first as a conduit for migrating cells to cross between the vehicle and recipient surface and second to shield adherent cells from destruction by mechanical shearing during handling and application. Quantitative analysis showed that topographic projections (columns) rather than recesses (pits) in the hydrogel surface achieved the highest efficiency of cell transfer. In order to address the crucial relevance of microbiological contamination to the success of wound grafting, the effect of iodine on several common bacterial pathogens was examined using an XTT+C(Q10) kinetic cell viability assay. Increasing concentrations of iodine initially stressed and after 0.5% v/v were subsequently bacteriocidal for Gram-negative Pseudomonas aeruginosa and Escherichia coli and Gram-positive Bacillus subtillis and Staphylococcus aureus. Slightly higher doses of iodine (approx 1-1.5% v/v) were required to kill HaCaT cells outright, but for both pro- and eukaryotes the major determinant of cytotoxicity was absolute dose rather than duration of exposure. Iodine delivered by the hydrogel at low concentration was bacteriostatic but not apparently cytotoxic to epithelial cells as measured by MTT end-point cell viability assay. Zone of inhibition studies confirmed that bacteriocidal quantities of neomycin, phenol red, and silver could also be delivered using the same hydrogel. This research suggests that grafting cell-based biological dressings to wounds using a topographically modified hydrogel dressing capable of simultaneous reducing the microbiological threat to a successful outcome may be a realistic clinical proposition.     

Mutants of bacteriophage T4 that produce infective fibreless particles

In wild type bacteriophage T4 the long tail fibres serve both in the initial attachment of the phage to its host and in the triggering of tail contraction. A two-step model for the control of triggering suggests that particles lacking the product of gene 9, which are also structurally fibreless, might be infective. This is shown to be the case, even though such phage do not plate on restrictive strains of bacteria. However, starting from phage carrying an amber mutation in gene 9 it is easy to isolate additional mutations which, under restrictive conditions, permit fibreless plating (pfp mutations). Three such pfp mutations, having also a temperature-sensitive phenotype, have been isolated and shown to map in genes coding for structural components of the baseplate. The mode of action of these pfp mutations is not clear, though they certainly destabilize the baseplate, thereby making triggering easier. The pfp mutations are effective only when in combination with an amber mutation in gene 9 and not with amber mutations in tail fibre genes, establishing the essentially inhibitory nature of the control of triggering exercised by gene 9 product.     

Effects of mcr restriction of methylated CpG islands of the L1 transposons during packaging and plating stages of mammalian genomic library construction.

The use of optimally methylation-tolerant mcrA- mcrB- strains has been shown to produce an over tenfold increase in the plating efficiencies of mammalian genomic libraries, compared to a superior conventional phage host strain LE392 which is mcrB+. However, there is an even more significant effect of mcr restriction. Amongst the recombinants recovered with an mcrB+ host, we have found that there is an additional 30-fold reduction in the frequencies of clones containing the heavily methylated 5'-CpG island sequences of both the human and rat L1 repetitive elements. The mcrA product was also found to restrict clones of these methylated genomic segments, but not as strongly as mcrB. However, the use of packaging extracts made from mcrA+ lysogens did not result in convincing reductions in the recoveries of these dispersed methylated elements. The magnitude of mcr restriction during plating due to methylated dispersed elements is sufficient to make a significant proportion of mammalian genomes unclonable from genomic libraries constructed previously using conventional mcr+ hosts.     

A sizing artifact of DNA restriction fragments with unusually long single-stranded termini.

The restriction endonucleases (ENases) BstNI (CCATGG) and EcoRII (CCATGG) both cleave DNA at the same time sequences, but only EcoRII produces 5-nucleotide (nt) cohesive ends and is inhibited by 5-methylation of the inner cytosine. The low-Mr fragments in digests of mouse DNA made with these two ENases exhibit different mobilities during agarose-gel electrophoresis. The difference in the mobilities of the BstNI and EcoRII fragments from mouse DNA was not due to closely spaced, differentially methylated sites, or to alternate mechanisms such as circularization of the long cohesive ends of the EcoRII fragments, or to residual bound protein. Rather, it was due to the unusually long 5-nt single-stranded (ss) ends of fragments produced by EcoRII digestion, since the slower mobility of the EcoRII fragments was abolished by treatment with ss-specific nuclease. Similar mobility differences between BstNI and EcoRII fragments which could be removed by ss nuclease were also observed in digests of simian virus 40 DNA.