Nickel binding and immunological properties of the C-terminal domain of the Helicobacter pylori GroES homologue (HspA)

Helicobacter pylori synthesizes a heat-shock protein of the GroES class. The gene encoding this protein (heat-shock protein A, HspA) was recently cloned and it was shown to be unique in structure. H. pylori HspA consists of two domains: the N-terminal domain (domain A) homologous with other GroES proteins, and a C-terminal domain (domain B) corresponding to 27 additional residues resembling a metal-binding domain. Various recombinant proteins consisting of the entire HspA polypeptide, the A domain, or the B domain were produced independently as proteins fused to maltose-binding protein (MBP). Comparison of the divalent cation binding properties of the various MBP and MBP-fused proteins allowed us to conclude that HspA binds nickel ions by means of its C-terminal domain. HspA exhibited a high and specific affinity for nickel ions in comparison with its affinity for other divalent cations (copper, zinc, cobalt). Equilibrium dialysis experiments revealed that MBP–HspA binds nickel ions with an apparent dissociation constant (K_d) of 1.8 μM and a stoichiometry of 1.9 ions per molecule. The analysis of the deduced HspA amino acid sequences encoded by 35 independent clinical isolates demonstrated the existence of two molecular variants of HspA, i.e. a major and a minor variant present in 89% and 11% of strains, respectively. The two variants differed from each other by the simultaneous substitution of seven amino acids within the B domain, whilst the A domain was highly conserved amongst all the HspA proteins (99–100% identity). On the basis of serological studies, the highly conserved A domain of HspA was found to be the immunodominant domain. Functional complementation experiments were performed to test the properties of the two HspA variants. When co-expressed together with the H. pylori urease gene cluster in Escherichia coli cells, the two HspA variant-encoding genes led to a fourfold increase in urease activity, demonstrating that HspA in H. pylori has a specialized function with regard to the nickel metalloenzyme urease.     

Non-Invasive Determination of Left Ventricular Workload in Patients with Aortic Stenosis Using Magnetic Resonance Imaging and Doppler Echocardiography

Early detection and accurate estimation of aortic stenosis (AS) severity are the most important predictors of successful long-term outcomes in patients. Current clinical parameters used for evaluation of the AS severity have several limitations including flow dependency. Estimation of AS severity is specifically challenging in patients with low-flow and low transvalvular pressure gradient conditions. A proper diagnosis in these patients needs a comprehensive evaluation of the left ventricle (LV) hemodynamic loads. This study has two objectives: (1) developing a lumped-parameter model to describe the ventricular-valvular-arterial interaction and to estimate the LV stroke work (SW); (2) introducing and validating a new index, the normalized stroke work (N-SW), to assess the global hemodynamic load imposed on the LV. N-SW represents the global hemodynamic load that the LV faces for each unit volume of blood ejected. The model uses a limited number of parameters which all can be measured non-invasively using current clinical imaging modalities. The model was first validated by comparing its calculated flow waveforms with the ones measured using Cardiovascular Magnetic Resonance (CMR) in 49 patients and 8 controls. A very good correlation and concordance were found throughout the cycle (median root mean square: 12.21 mL/s) and between the peak values (r = 0.98; SEE = 0.001, p<0.001). The model was then used to determine SW using the parameters measured with transthoracic Doppler-echocardiography (TTE) and CMR. N-SW showed very good correlations with a previously-validated index of global hemodynamic load, the valvular arterial impedance (), using data from both imaging modalities (TTE: r = 0.82, SEE = 0.01, p<0.001; CMR: r = 0.74, SEE = 0.01, p<0.001). Furthermore, unlike , N-SW was almost independent from variations in the flow rate. This study suggests that considering N-SW may provide incremental diagnostic and prognostic information, beyond what standard indices of stenosis severity and provide, particularly in patients with low LV outflow.     

Inter‐oceanic variation in patterns of host‐associated divergence in a seabird ectoparasite

Aim Parasites with global distributions and wide host spectra provide excellent models for exploring the factors that drive parasite diversification. Here, we tested the relative force of host and geography in shaping population structure of a widely distributed and common ectoparasite of colonial seabirds, the tick Ixodes uriae. Location Two natural geographic replicates of the system: numerous seabird colonies of the North Pacific and North Atlantic Ocean basins. Methods Using eight microsatellite markers and tick samples from a suite of multi‐specific seabird colonies, we examined tick population structure in the North Pacific and compare patterns of diversity and structure to those in the Atlantic basin. Analyses included population genetic estimations of diversity and population differentiation, exploratory multivariate analyses, and Bayesian clustering approaches. These different analyses explicitly took into account both the geographic distance among colonies and host use by the tick. Results Overall, little geographic structure was observed among Pacific tick populations. However, host‐related genetic differentiation was evident, but was variable among host types and lower than in the North Atlantic. Main conclusions Tick population structure is concordant with the genetic structure observed in seabird host species within each ocean basin, where seabird populations tend to be less structured in the North Pacific than in the North Atlantic. Reduced tick genetic structure in the North Pacific suggests that host movement among colonies, and thus tick dispersal, is higher in this region. In addition to information on parasite diversity and gene flow, our findings raise interesting questions about the subtle ways that host behaviour, distribution and phylogeographic history shape the genetics of associated parasites across geographic landscapes.     

Settlement pattern of tortoises translocated into the wild: a key to evaluate population reinforcement success

A lack of long-term monitoring often impedes the evaluation of translocation used to reinforce populations. Crucial questions regarding the exact timing and place of possible settlement remain unanswered. To examine these issues we radio-tracked during three years 24 tortoises (Testudo hermanni hermanni) released to reinforce a resident population impacted by fire. Individuals from the resident population (N = 20) and from a distant control population (N = 11) were also radio-tracked. More than 11,000 fixes were collected, enabling us to precisely describe movement patterns. Most translocated tortoises first dispersed (> 500 m to > 3000 m away) in a random direction and sometimes crossed unfavorable areas. Later, a marked shift in movement pattern, from a relatively unidirectional course to multidirectional displacements indicated settlement. Movement patterns of translocated and resident individuals became undistinguishable after settlement. Most individuals settled during the first year after release but several settled in the second year. Mean annual survival rate (> 85%) remained within the range of the species but was lower compared to the resident (93%) and control tortoises (100%). Overall, most translocated individuals (~ 63%) settled and adapted well to their novel environment. This result is essential regarding current controversies that are unfounded and that limit conservation translocations. Yet, translocation sites should be large enough and/or surrounded by secondary favorable areas to limit the mortality associated with dispersal in hazardous environments. Large numbers of individuals rescued during urbanization works may easily supply conservation translocations to reinforce fragile populations.     

Oral microbiomes from hunter‐gatherers and traditional farmers reveal shifts in commensal balance and pathogen load linked to diet

Maladaptation to modern diets has been implicated in several chronic disorders. Given the higher prevalence of disease such as dental caries and chronic gum diseases in industrialized societies, we sought to investigate the impact of different subsistence strategies on oral health and physiology, as documented by the oral microbiome. To control for confounding variables such as environment and host genetics, we sampled saliva from three pairs of populations of hunter‐gatherers and traditional farmers living in close proximity in the Philippines. Deep shotgun sequencing of salivary DNA generated high‐coverage microbiomes along with human genomes. Comparing these microbiomes with publicly available data from individuals living on a Western diet revealed that abundance ratios of core species were significantly correlated with subsistence strategy, with hunter‐gatherers and Westerners occupying either end of a gradient of Neisseria against Haemophilus, and traditional farmers falling in between. Species found preferentially in hunter‐gatherers included microbes often considered as oral pathogens, despite their hosts' apparent good oral health. Discriminant analysis of gene functions revealed vitamin B5 autotrophy and urease‐mediated pH regulation as candidate adaptations of the microbiome to the hunter‐gatherer and Western diets, respectively. These results suggest that major transitions in diet selected for different communities of commensals and likely played a role in the emergence of modern oral pathogens.     

Large scale Agriotes spp. click beetle (Coleoptera: Elateridae) invasion of crop land from field margin reservoirs

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Human plasma protein <Emphasis Type="Italic">N</Emphasis>-glycosylation

Glycosylation is the most abundant and complex protein modification, and can have a profound structural and functional effect on the conjugate. The oligosaccharide fraction is recognized to be involved in multiple biological processes, and to affect proteins physical properties, and has consequentially been labeled a critical quality attribute of biopharmaceuticals. Additionally, due to recent advances in analytical methods and analysis software, glycosylation is targeted in the search for disease biomarkers for early diagnosis and patient stratification. Biofluids such as saliva, serum or plasma are of great use in this regard, as they are easily accessible and can provide relevant glycosylation information. Thus, as the assessment of protein glycosylation is becoming a major element in clinical and biopharmaceutical research, this review aims to convey the current state of knowledge on the N-glycosylation of the major plasma glycoproteins alpha-1-acid glycoprotein, alpha-1-antitrypsin, alpha-1B-glycoprotein, alpha-2-HS-glycoprotein, alpha-2-macroglobulin, antithrombin-III, apolipoprotein B-100, apolipoprotein D, apolipoprotein F, beta-2-glycoprotein 1, ceruloplasmin, fibrinogen, immunoglobulin (Ig) A, IgG, IgM, haptoglobin, hemopexin, histidine-rich glycoprotein, kininogen-1, serotransferrin, vitronectin, and zinc-alpha-2-glycoprotein. In addition, the less abundant immunoglobulins D and E are included because of their major relevance in immunology and biopharmaceutical research. Where available, the glycosylation is described in a site-specific manner. In the discussion, we put the glycosylation of individual proteins into perspective and speculate how the individual proteins may contribute to a total plasma N-glycosylation profile determined at the released glycan level.     

ISOLATION AND CHARACTERIZATION OF SIX cDNAs INVOLVED IN CARBON METABOLISM IN LAMINARIA DIGITATA (PHAEOPHYCEAE)

An expressed sequence tag (EST) approach was used to retrieve cDNA clones involved in carbon metabolism in Laminaria digitata Lamouroux. Six partial open reading frames were identified, respectively encoding an α-type carbonic anhydrase (CA), glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH), phosphoglycerate kinase (PGK), glycolate oxidase (GLO), and GDP-4-keto-6-d-mannose-3,5-epimerase-4-reductase, also known as fucose synthase (FS). These enzymes were further characterized through Southern blot analyses, amino acid sequence comparisons, and patterns of expression. In contrast to the other genes, which are expressed in both the gametophytic and sporophytic generations of L. digitata, the α-type carbonic anhydrase messenger RNA was shown by RT-PCR and northern blot analyses to be present in the gametophytes only. The evolutionary relationships of these genes and their interest as molecular tools for investigating carbon fluxes in brown algae are discussed.     

How threatened are large branchiopods (Crustacea,Branchiopoda) in the Iberian Peninsula?

Aquatic macroinvertebrates in drainage ditches may alter rates of nutrient cycling and decomposition of organic matter but have not been accounted for in studies of ditch biogeochemistry. We collected sediment cores from four pairs of field (intermittent) and collection (perennial) ditches on Maryland’s Eastern Shore monthly from March 2011 to February 2012 to determine how taxonomic and functional group composition varies among different ditch types. We identified 138 taxa and assigned them to functional groups according to trophic position and modes of burrowing. There was no difference in mean abundance of invertebrates (5821 ind./m²) between seasons or types of ditches, and species richness peaked in winter (20 taxa/site) compared to other seasons (15 taxa/site), but did not vary between ditch types. Assemblage composition differed between field and collection ditches, but functional group composition did not. Field ditches flow intermittently which may limit the assemblage to early colonists and taxa adapted to survive desiccation. The benthic macroinvertebrate assemblage was dominated by the collector–gatherer functional feeding group (83.6%) and burrowing taxa (97.1%). Bioturbation by burrowing macroinvertebrates is likely an important process contributing to ecosystem-scale functions of drainage ditches, including regulation of biogeochemical processes occurring at the sediment–water interface.