Apparent absence of blood parasites in Winter Wrens in British Columbia

ABSTRACT.   To investigate the possible effect of elevation on the prevalence of hematozoa infection, we collected blood smears from a population of Winter Wrens ( Troglogytes troglodytes ) in the Coast Mountains of southwestern British Columbia, Canada. We sampled 119 Winter Wrens, including 88 at low elevation (0–500 m) and 31 at high elevation (900–1100 m) sites. In addition, five other species, including Swainson's Thrush ( Catharus ustulatus ; N = 12), Dark-eyed Junco ( Junco hyemalis ; N = 11), Hermit Thrush ( Catharus guttatus ; N = 8), Varied Thrush ( Ixoreus naevius ; N = 4), and American Robin ( Turdus migratorius ; N = 4), were sampled. No Winter Wrens were infected by blood parasites. Among the other species, Haemoproteus infection was detected in one Varied Thrush (25%) and five Swainson's Thrushes (42%). Thus, despite the occurrence of infection in sympatrically breeding species, blood parasites are apparently absent or occur at extremely low prevalence in Winter Wrens in our study area. The presence of hematozoa in European populations of Winter Wrens, combined with an abundance of vector species in our study area, suggest population-level resistance to infection. Further study is needed to determine the specific mechanisms involved in the apparent lack of infection in our study population.     

The period gene Thr-Gly polymorphism in Australian and African Drosophila melanogaster populations: implications for selection

The period gene is a key regulator of biological rhythmicity in Drosophila melanogaster. The central part of the gene encodes a dipeptide Thr-Gly repeat that has been implicated in the evolution of both circadian and ultradian rhythms. We have previously observed that length variation in the repeat follows a latitudinal cline in Europe and North Africa, so we have sought to extend this observation to the southern hemisphere. We observe a parallel cline in Australia for one of the two major length variants and find higher levels of some Thr-Gly length variants, particularly at the tropical latitudes, that are extremely rare in Europe. In addition we examined >40 haplotypes from sub-Saharan Africa and find a very different and far more variable profile of Thr-Gly sequences. Statistical analysis of the periodicity and codon content of the repeat from all three continents reveals a possible mechanism that may explain how the repeat initially arose in the ancestors of the D. melanogaster subgroup of species. Our results further reinforce the view that thermal selection may have contributed to shaping the continental patterns of Thr-Gly variability.     

Crystal structure of the human TRPV2 channel ankyrin repeat domain

TRPV channels are important polymodal integrators of noxious stimuli mediating thermosensation and nociception. An ankyrin repeat domain (ARD), which is a common protein-protein recognition domain, is conserved in the N-terminal intracellular domain of all TRPV channels and predicted to contain three to four ankyrin repeats. Here we report the first structure from the TRPV channel subfamily, a 1.7 A resolution crystal structure of the human TRPV2 ARD. Our crystal structure reveals a six ankyrin repeat stack with multiple insertions in each repeat generating several unique features compared with a canonical ARD. The surface typically used for ligand recognition, the ankyrin groove, contains extended loops with an exposed hydrophobic patch and a prominent kink resulting from a large rotational shift of the last two repeats. The TRPV2 ARD provides the first structural insight into a domain that coordinates nociceptive sensory transduction and is likely to be a prototype for other TRPV channel ARDs.     

Helminth assemblages of the turtle Emydura macquarii (Pleurodira: Chelidae) Queensland, Australia

The helminth fauna of 76 Emydura macquarii from 3 river systems in central and northern Queensland was examined. Eleven species were found, including 2 nematodes, 6 trematodes, 1 aspidogastrean, 1 cestode, and 1 monogenean. Analysis of helminth diversity showed that the Fitzroy and Ross River turtles had communities of comparable diversity, but the helminth communities in Proserpine River turtles were much less diverse. The helminth communities in all localities were dominated by trematodes. Polystomoides australiensis was the most prevalent, being found in 60% of the Ross River turtles, 57% of the Fitzroy River turtles, and 46% of the Proserpine River turtles. Notopronocephalus peekayi was the most abundant species, with mean abundances of 5.9 in the Ross River turtles and 9.8 in the Fitzroy River turtles. Species richness, Simpson's Reciprocal Index, was highest, 4.68, for the Ross River helminth community, Sorensen's Qualitative Index showed 95% similarity between the Ross River and Fitzroy River communities, although Sorensen's Quantitative Index indicated only 35% similarity between the 2 sites. Host feeding patterns are likely the most important factor affecting species richness of the helminth infracommunities, as the majority of helminth species are transmitted by food-web interactions involving intermediate hosts.     

Secondary osteon and Haversian canal dimensions as behavioral indicators

Variation in the size of structures within mature cortical bone is relevant to our understanding of apparent differences between human samples, and it is relevant to the development of histologically based age-estimation methods. It was proposed that variation may reflect effects of physical activity, through biomechanical and/or metabolic mechanisms. If these factors are local, femoral osteon area (On.Ar) should be more histologically variable than On.Ar in ribs. Ribs should show a higher variation in Haversian canal area (H.Ar) if they are sites of more remodeling activity and hence of arrested refilling of secondary osteons at time of death. This study compares On.Ar and H.Ar of secondary osteons from femora (15) and ribs (29) from 44 Holocene (Later Stone Age) foragers from South Africa (M = 19, F = 25) to values from paired femora and ribs from historic samples (Spitalfields and St. Thomas, 20 pairs from each). Fixed-effects analysis of variance demonstrates rib On.Ar to be significantly smaller than femur, but with no sex or age effects. The femur-to-rib On.Ar ratio is lower for the Holocene foragers than for the two modern samples because of relatively large rib On.Ar. Femora and ribs from the same skeleton normally show femoral On.Ar larger than rib On.Ar (37/44 pairs). Mean femoral values of On.Ar are more diverse than rib On.Ar values, but within-sample coefficients of variation are similar. Values for H.Ar are highly variable and do not reflect anatomical site, age, sex, or population effects. The patterning of osteon size does not appear to be linked to physical activity or to different rates of metabolic activity within the skeleton, at least not in a straightforward way.     

Engineering of an anti-epidermal growth factor receptor antibody to single chain format and labeling by Sortase A-mediated protein ligation

Sortase-mediated protein ligation is a biological covalent conjugation system developed from the enzymatic cell wall display mechanism found in Staphylococcus aureus. This three-component system requires: (i) purified Sortase A (SrtA) enzyme; (ii) a substrate containing the LPXTG peptide recognition sequence; and (iii) an oligo-glycine acceptor molecule. We describe cloning of the single-chain antibody sc528, which binds to the extracellular domain of the epidermal growth factor receptor (EGFR), from the parental monoclonal antibody and incorporation of a LPETGG tag sequence. Utilizing recombinant SrtA, we demonstrate successful incorporation of biotin from GGG-biotin onto sc528. EGFR is an important cancer target and is over-expressed in human tumor tissues and cancer lines, such as the A431 epithelial carcinoma cells. SrtA-biotinylated sc528 specifically bound EGFR expressed on A431 cells, but not negative control lines. Similarly, when sc528 was labeled with fluorescein we observed antigen-specific labeling. The ability to introduce functionality into recombinant antibodies in a controlled, site-specific manner has applications in experimental, diagnostic, and potentially clinical settings. For example, we demonstrate addition of all three reaction components in situ within a biosensor flow cell, resulting in oriented covalent capture and presentation of sc528, and determination of precise affinities for the antibody-receptor interaction.     

Bioleaching in brackish waters—effect of chloride ions on the acidophile population and proteomes of model species

High concentrations of chloride ions inhibit the growth of acidophilic microorganisms used in biomining, a problem particularly relevant to Western Australian and Chilean biomining operations. Despite this, little is known about the mechanisms acidophiles adopt in order to tolerate high chloride ion concentrations. This study aimed to investigate the impact of increasing concentrations of chloride ions on the population dynamics of a mixed culture during pyrite bioleaching and apply proteomics to elucidate how two species from this mixed culture alter their proteomes under chloride stress. A mixture consisting of well-known biomining microorganisms and an enrichment culture obtained from an acidic saline drain were tested for their ability to bioleach pyrite in the presence of 0, 3.5, 7, and 20 g L⁻¹ NaCl. Microorganisms from the enrichment culture were found to out-compete the known biomining microorganisms, independent of the chloride ion concentration. The proteomes of the Gram-positive acidophile Acidimicrobium ferrooxidans and the Gram-negative acidophile Acidithiobacillus caldus grown in the presence or absence of chloride ions were investigated. Analysis of differential expression showed that acidophilic microorganisms adopted several changes in their proteomes in the presence of chloride ions, suggesting the following strategies to combat the NaCl stress: adaptation of the cell membrane, the accumulation of amino acids possibly as a form of osmoprotectant, and the expression of a YceI family protein involved in acid and osmotic-related stress.