Phylogenetic and morphological characterisation of the green algae infesting blue mussel Mytilus edulis in the North and South Atlantic oceans

Blue mussels Mytilus edulis with shell deformations and green pustules containing parasitic algae were collected at 3 coastal sites (Bur?y, Norway; Bockholm, Denmark; Goose Green, Falkland Islands). A comparative study, including mussel histopathology, algal morphology, ultrastructure and phylogenetic position was performed. Green pustules were mainly located in the posterior portion of the mantle and gonad tissues and the posterior adductor muscle. Electron microscopy confirmed the presence of algal cells with similar morphology to Coccomyxa parasitica. Algae were oval shaped with a single nucleus and chloroplast, 1 or 2 mitochondria and a dense granular cytoplasm with a lipid inclusion body, Golgi apparatus and small vesicles. Partial small subunit (SSU) rRNA phylogeny confirmed the inclusion of parasitic algae into the Coccomyxa clade. However, the sequence identity between almost full SSU rRNA sequences of parasitic algae and others in this clade yielded an unexpected result. Green algae from mussels were distant from C. parasitica Culture Collection of Algae and Protozoa (CCAP) strain 216/18 (94% identity), but very similar (99% identity) to C. glaronensis (a lichen endosymbiont) and green endophytes from the tree Ginkgo biloba. The CCAP strain 216/18 was a sister sequence to Nannochloris algae, far from the Coccomyxa clade. These results suggest a misidentification or outgrowth of the original CCAP strain 216/18 by a different 'Nannochloris-like' trebouxiophycean organism. In contrast, our sequences directly obtained from infested mussels could represent the true C. parasitica responsible for the green pustules in blue mussels.     

The imprint of Quaternary glaciers on the present-day distribution of the obligate groundwater amphipod Niphargus virei (Niphargidae)

Aim A plausible yet untested biogeographical scenario suggests that Quaternary glaciers shaped the present‐day distribution of the groundwater amphipod Niphargus virei. This study was designed to test two hypotheses pertaining to this scenario: (1) the probability of occurrence of N. virei in ice‐free areas decreases in the vicinity of the Würm glacier; and (2) dispersal is sufficiently low for the historical record of glacial effects to persist over time. Location The study area was located in the southern Jura Mountains, France. Methods A total of 497 sites were sampled to ascertain the distribution of N. virei in the southern Jura. Amplified fragment length polymorphism was analysed from a subset of 24 sites. The relationships between the probability of occurrence of N. virei and distance to the Würm glacier or elevation were investigated using a logistic regression. Spatial autocorrelation analyses were performed on both the residuals of the logistic regression and genetic distance to test the significance of dispersal and barriers to post‐glacial recolonization. The influence of catchment boundaries as barriers to dispersal was examined using different neighbouring relationships between sites. We tested the statistical significance of the reduction in deviance and gain in precision of an autologistic regression that took into consideration the influence of dispersal constraints on the distribution of N. virei. Results Niphargus virei rarely occurred in formerly glaciated areas, and its probability of occurrence in ice‐free areas decreased in the vicinity of the Würm glacier. Combined autocorrelation analyses of spatial distribution and spatial genetic structure showed that: (1) the distance at which spatial autocorrelation was no longer significantly positive did not exceed 16 km; (2) genetic differentiation fitted a model of isolation by distance; and (3) catchment boundaries acted as barriers to dispersal. The autologistic regression with dispersal constraints significantly increased our capacity to predict the distribution of N. virei. Maps of probabilities of occurrence suggested that post‐glacial recolonization was impeded by the extension of glacial outwash. Main conclusions The present distribution of N. virei in southern Jura is probably the result of a historical range reduction driven by glaciation coupled with restricted dispersal and isolation by distance.     

Finite-size transitions in complex membranes

The lipid-raft hypothesis postulates that cell membranes possess some degree of lateral organization. The hypothesis has attracted much attention while remaining controversial, with an underlying mechanism that remains elusive. One idea that supports rafts relies on the membrane lying near a critical point. Although supported by experimental evidence, holding a many-component membrane at criticality requires a delicate tuning of all components—a daunting task. Here, we propose a coherent framework to reconcile critical behavior and lipid regulation. Using a lattice model, we show that lipid regulation of a complex membrane, i.e., allowing composition to fluctuate based on relative chemical potentials, can lead to critical behavior. The results are robust against specific values of the chemical potentials. Instead of a conventional transition point, criticality is observed over a large temperature range. This surprising behavior arises from finite-size effects, causing nonequivalent time and space averages. The instantaneous lipid distribution effectively develops a translational symmetry, which we relate to long-wavelength Goldstone modes. The framework is robust and reproduces important experimental trends; membrane-demixing temperature closely follows cell-growth temperature. It also ensures criticality of fixed-composition extracts, such as giant plasma membrane vesicles. Our clear picture provides a strong argument in favor of the critical-membrane hypothesis, without the need for specific sensing mechanisms.     

DspA/E, a type III effector essential for Erwinia amylovora pathogenicity and growth in planta, induces cell death in host apple and nonhost tobacco plants

Erwinia amylovora is responsible for fire blight, a necrotic disease of apples and pears. E. amylovora relies on a type III secretion system (TTSS) to induce disease on hosts and hypersensitive response (HR) on nonhost plants. The DspA/E protein is essential for E. amylovora pathogenicity and is secreted via the TTSS in vitro. DspA/E belongs to a type III effector family that is conserved in several phytopathogenic bacteria. In E. amylovora, DspA/E has been implicated in the generation of an oxidative stress during disease and the suppression of callose deposition. We investigated the fate of DspA/E in planta. DspA/E delivered artificially to apple or tobacco cells by agroinfection induced necrotic symptoms, indicating that DspA/E was probably injected via the TTSS. We confirmed that DspA/E acts as a major cell-death inducer during disease and HR, because the dspA/E mutant is severely impaired in its ability to induce electrolyte leakage in apple and tobacco leaves. Expression of the defense marker gene PR1 was delayed when dspA/E was transiently expressed in tobacco, suggesting that DspA/E-mediated necrosis may be associated with an alteration of defense responses.     

Microstructured liposome array

Conversion of a DNA chip to a nanocapsule array was performed by grafting on a liposome an oligonucleotide complementary to an oligonucleotide bound to the array. Each liposome may be loaded by a soluble molecule or may present a hydrophobic or amphiphilic molecule inserted in its wall. To detect liposomes on the chip, we used fluorescent dyes encapsulated in the liposome internal volume or fluorescent lipids. We observed that an oligonucleotide-grafted liposome containing a defined dye specifically accumulated on the area where its complementary oligonucleotide had been spotted on the array. The virtually unlimited amount of addresses allows the specific binding of large amounts of liposomes in one single batch.     

Structural characterization of CA1462, the <Emphasis Type="Italic">Candida albicans</Emphasis> thiamine pyrophosphokinase

Background  

In search of new antifungal targets of potential interest for pharmaceutical companies, we initiated a comparative genomics study to identify the most promising protein-coding genes in fungal genomes. One criterion was the protein sequence conservation between reference pathogenic genomes. A second criterion was that the corresponding gene in Saccharomyces cerevisiae should be essential. Since thiamine pyrophosphate is an essential product involved in a variety of metabolic pathways, proteins responsible for its production satisfied these two criteria.     

Synthesis and characterization of radiopaque iodine-containing degradable PVA hydrogels

Poly (vinyl alcohol) (PVA) hydrogels are highly attractive for biomedical applications, especially for controlled release of drugs and proteins. Recently, degradable PVA hydrogels have been described, having the advantage that the material disappears over time from the implantation site. Herein, we report the synthesis of radiopaque degradable PVA, which gives a further advantage that the position of the hydrogel can precisely be determined by X-ray fluoroscopy. Radiopacity has been introduced by replacing 0.5% of the pendent alcohol groups on the PVA with 4-iodobenzoylchloride. This level of substitution rendered the polymer adequately radiopaque. The subsequent modification of 0.8% of the pendent hydroxyl groups with an ester acrylate functional group allowed for cross-linking of the macromers. The radiopaque hydrogels degraded over a time span of 140 days. Rheology data suggested that the macromer solutions were appropriate for injection.     

In vivo activation of midbrain dopamine neurons via sensitized, high-affinity alpha 6 nicotinic acetylcholine receptors

Alpha6-containing (alpha6*) nicotinic ACh receptors (nAChRs) are selectively expressed in dopamine (DA) neurons and participate in cholinergic transmission. We generated and studied mice with gain-of-function alpha6* nAChRs, which isolate and amplify cholinergic control of DA transmission. In contrast to gene knockouts or pharmacological blockers, which show necessity, we show that activating alpha6* nAChRs and DA neurons is sufficient to cause locomotor hyperactivity. alpha6(L9'S) mice are hyperactive in their home cage and fail to habituate to a novel environment. Selective activation of alpha6* nAChRs with low doses of nicotine, by stimulating DA but not GABA neurons, exaggerates these phenotypes and produces a hyperdopaminergic state in vivo. Experiments with additional nicotinic drugs show that altering agonist efficacy at alpha6* provides fine tuning of DA release and locomotor responses. alpha6*-specific agonists or antagonists may, by targeting endogenous cholinergic mechanisms in midbrain or striatum, provide a method for manipulating DA transmission in neural disorders.