Comparative structural analysis of the calcium free and bound states of the calcium regulatory protein calbindin D9K

The solution structure of apo calbindin D9K, a member of the calmodulin superfamily of calcium-binding regulatory proteins, has been investigated by 1H nuclear magnetic resonance spectroscopy and the results compared with a corresponding study of the calcium-loaded protein. On the basis of complete sequence-specific assignments, characteristic patterns of short proton-proton distances have been identified in two-dimensional nuclear Overhauser effect spectra, allowing the elements of secondary structure to be determined. It is found that four helices and a short section of antiparallel beta-sheet are present regardless of the calcium content of the protein. In addition, a preliminary analysis of the long-range nuclear Overhauser effects shows that the global folding patterns are the same and that the tertiary structures of the apo protein is very similar to that of the calcium-loaded protein. These results are in stark contrast to a number of very substantial changes in 1H chemical shift. Preliminary studies of protein dynamics show some very large differences in flexibility and internal mobility. This suggests that protein dynamics may play a role more important than was initially realized in the function of calbindin D9K and other homologous calcium-binding regulatory proteins.     

Selection of the conodont Idiognathodus simulator (Ellison) as the event marker for the base of the global Gzhelian Stage (Upper Pennsylvanian, Carboniferous)

Studies carried out for more than 10 years by the Task Group to establish GSSPs at the base of the Moscovian–Kasimovian and Kasimovian–Gzhelian boundaries have resulted in the proposal that the level at which the conodont species Idiognathodus simulator (Ellison, 1941) first appears be selected to mark the base of the Gzhelian Stage. This expands this eastern European chronostratigraphic unit to a global scale.I. simulator (sensu Barrick et al., 2008) has been identified so far in Midcontinent and eastern North America, the Moscow and Donets basins and southern Urals of eastern Europe, and in south-central China. Correlation of this level based on this species and other conodont species can be reinforced in some areas by ammonoid and fusulinid data.     

Feeding ecology and specialist diet of critically endangered Ridgway's Hawks

Ridgway's Hawks (Buteo ridgwayi) are critically endangered forest raptors endemic to the Caribbean island of Hispaniola, with ～100 pairs remaining in the world. The species is ecologically little known yet such studies are important for understanding critical habitat needs and population dynamics. We studied the provisioning behavior of adults at 22 nests on the northeast coast of the Dominican Republic from 2005 to 2008. Mean brood size was 1.80 ± 0.45, and the mean number of fledglings per nest was 1.10 ± 0.97. We found that 80% of the prey items delivered to nestlings were reptiles, with lizards accounting for 65% of the prey and those in the genus Celestus accounting for nearly 35% of prey. Other prey items included snakes (14%), rats (9%), and smaller proportions of birds, frogs, and centipedes. The number of prey items and amount of biomass delivered to nestlings did not vary with brood size, but adults delivered more prey to 3‐ to 5‐week‐old nestlings and more biomass to 5‐week‐old nestlings. Food delivery rates did not differ between successful or failed nests, suggesting that food availability did not influence nest outcome. Given that most prey items delivered to nestlings in our study were reptiles, conservation strategies developed for Ridgway's Hawks (e.g., translocations and habitat conservation) should take into consideration their specialist reptile diet.     

Quantitative secretome and glycome of primary human adipocytes during insulin resistance

Adipose tissue is both an energy storage depot and an endocrine organ. The impaired regulation of the secreted proteins of adipose tissue, known as adipocytokines, observed during obesity contributes to the onset of whole-body insulin resistance and the pathobiology of type 2 diabetes mellitus (T2DM). In addition, the global elevation of the intracellular glycosylation of proteins by O-linked β-N-acetylglucosamine (O-GlcNAc) via either genetic or pharmacological methods is sufficient to induce insulin resistance in both cultured cells and animal models. The elevation of global O-GlcNAc levels is associated with the altered expression of many adipocytokines. We have previously characterized the rodent adipocyte secretome during insulin sensitive and insulin resistant conditions. Here, we characterize and quantify the secretome and glycome of primary human adipocytes during insulin responsive and insulin resistant conditions generated by the classical method of hyperglycemia and hyperinsulinemia or by the pharmacological manipulation of O-GlcNAc levels. Using a proteomic approach, we identify 190 secreted proteins and report a total of 20 up-regulated and 6 down-regulated proteins that are detected in both insulin resistant conditions. Moreover, we apply glycomic techniques to examine (1) the sites of N-glycosylation on secreted proteins, (2) the structures of complex N- and O-glycans, and (3) the relative abundance of complex N- and O-glycans structures in insulin responsive and insulin resistant conditions. We identify 91 N-glycosylation sites derived from 51 secreted proteins, as well as 155 and 29 released N- and O-glycans respectively. We go on to quantify many of the N- and O-glycan structures between insulin responsive and insulin resistance conditions demonstrating no significant changes in complex glycosylation in the time frame for the induction of insulin resistance. Thus, our data support that the O-GlcNAc modification is involved in the regulation of adipocytokine secretion upon the induction of insulin resistance in human adipocytes.     

Using high‐resolution multi‐spectral imagery to estimate habitat complexity in open‐canopy forests: can we predict ant community patterns?

The structure and composition of arthropod assemblages are strongly associated with habitat complexity. Accurate, time efficient estimates of habitat complexity may provide insights for biodiversity management in natural systems. We obtained high-resolution (0.7 m pixel) multi-spectral aerial imagery of National Parks 20 km north and 20 km south of Sydney, Australia. We explored both the Normalised Difference Vegetation Index (NDVI) and the standard deviation of reflectance in the near-infrared spectrum (stdevR_NIR) as indicators of low and high habitat complexity in sandstone forests north of Sydney. We then tested described predictions of ant community patterns (based on a previous study) using sites selected from high-resolution multi-spectral imagery in sandstone forests south of Sydney. Ground-scored habitat complexity was positively correlated with NDVIs and, to a lesser extent, stdevR_NIR values in sandstone forests north of Sydney. As predicted, ant species richness was negatively correlated with NDVIs in forests to the south of Sydney. Also, ant species composition was different in areas with contrasting NDVIs. The ant species driving composition differences responded to habitat complexity in a similar way in forests to the north, and south, of Sydney. The strong association we detected between NDVIs and habitat complexity, most likely reflects the relatively exposed nature of the vegetative layers in the forests we sampled. Remote sensing, integrated with quantitative research testing predictive faunal responses to vegetation structure and biomass at landscape scales, may provide efficient means of estimating biodiversity for management in particular habitats.     

The F1-ATPase from Streptococcus cremoris: isolation, purification and partial characterization

1. The F1-ATPase from the plasma membrane of Streptococcus cremoris HA was released by low ionic shock wash and purified by gel filtration and ion exchange chromatography. 2. The specific activity of the purified F1-ATPase was 25.8 mumol Pi/mg protein/min. 3. Km for ATP was 0.80 mM, and Ki for ADP as a competetive inhibitor 0.40 mM. 4. The purified F1-ATPase consisted of five subunits, alpha, beta, gamma, delta and epsilon, with molecular masses of 47.0, 45.0, 29.5, 22.0 and 13.0 kDa, respectively. 5. The isoelectric point of the enzyme complex was found to be 4.4.     

Disease-driven reduction in human mobility influences human-mosquito contacts and dengue transmission dynamics

Heterogeneous exposure to mosquitoes determines an individual’s contribution to vector-borne pathogen transmission. Particularly for dengue virus (DENV), there is a major difficulty in quantifying human-vector contacts due to the unknown coupled effect of key heterogeneities. To test the hypothesis that the reduction of human out-of-home mobility due to dengue illness will significantly influence population-level dynamics and the structure of DENV transmission chains, we extended an existing modeling framework to include social structure, disease-driven mobility reductions, and heterogeneous transmissibility from different infectious groups. Compared to a baseline model, naïve to human pre-symptomatic infectiousness and disease-driven mobility changes, a model including both parameters predicted an increase of 37% in the probability of a DENV outbreak occurring; a model including mobility change alone predicted a 15.5% increase compared to the baseline model. At the individual level, models including mobility change led to a reduction of the importance of out-of-home onward transmission (R, the fraction of secondary cases predicted to be generated by an individual) by symptomatic individuals (up to -62%) at the expense of an increase in the relevance of their home (up to +40%). An individual’s positive contribution to R could be predicted by a GAM including a non-linear interaction between an individual’s biting suitability and the number of mosquitoes in their home (>10 mosquitoes and 0.6 individual attractiveness significantly increased R). We conclude that the complex fabric of social relationships and differential behavioral response to dengue illness cause the fraction of symptomatic DENV infections to concentrate transmission in specific locations, whereas asymptomatic carriers (including individuals in their pre-symptomatic period) move the virus throughout the landscape. Our findings point to the difficulty of focusing vector control interventions reactively on the home of symptomatic individuals, as this approach will fail to contain virus propagation by visitors to their house and asymptomatic carriers.     

Drosophila do not skew offspring sex ratio as predicted by trivers and willard (Diptera: Drosophilidae)

Based on both previously published literature and results reported here, it appears thatDrosophila melanogaster meet the explicit assumptions of the Trivers and Willard offspring sex allocation model. However, contrary to the model's predictions, offspring sex ratio was not significantly affected when we manipulated factors that influence offspring quality. We suggest that contrary to implicit predictions of offspring sex ratio models,Drosophila may lack the genetic plasticity to readily alter sex ratio.