TccP is an enterohaemorrhagic Escherichia coli O157:H7 type III effector protein that couples Tir to the actin-cytoskeleton

Subversion of host cell actin microfilaments is the hallmark of enterohaemorrhagic (EHEC) and enteropathogenic (EPEC) Escherichia coli infections. Both pathogens translocate the trans-membrane receptor protein-translocated intimin receptor (Tir), which links the extracellular bacterium to the cell cytoskeleton. While both converge on neural Wiskott-Aldrich syndrome protein (N-WASP), Tir-mediated actin accretion by EPEC and EHEC differ in that Tir(EPEC) requires both tyrosine phosphorylation and the host adaptor protein Nck, whereas Tir(EHEC) is not phosphorylated and utilizes an unidentified linker. Here we report the identification of Tir-cytoskeleton coupling protein (TccP), a novel EHEC effector that displays an Nck-like coupling activity following translocation into host cells. A tccP mutant did not affect Tir translocation and focusing but failed to recruit alpha-actinin, Arp3, N-WASP and actin to the site of bacterial adhesion. When expressed in EPEC, bacterial-derived TccP restored actin polymerization activity following infection of an Nck-deficient cell line. TccP has a similar biological activity on infected human intestinal explants ex vivo. Purified TccP activates N-WASP stimulating, in the presence of Arp2/3, actin polymerization in vitro. These results show that EHEC translocates both its own receptor (Tir) and an Nck-like protein (TccP) to facilitate actin polymerization.     

Testing the usefulness of a simple automatic method for particles abundance and size determination to derive cost-effective biological indicators in large monitoring networks

A simple automatic method has been tested for biological quality assessment in coastal and estuarine waters. Samples collected in routine campaigns along the Basque coast (northern Spain) were analysed by both, a traditional microscopy technique (Utermöhl) and the flow cytometer and microscope (FlowCAM). The FlowCAM was operated in AutoImage mode (i.e. without fluorescence measurements). Two different FlowCAM procedures were carried out in two campaigns separately (2005 and 2008). These differed mainly in the lower size-threshold of the particles sampled: 10 μm in 2005, and 4 μm in 2008. The degree of divergence was evaluated, recognizing that the Utermöhl method and the FlowCAM (as employed here) count differently (i.e. phytoplankton cells and particles, respectively). In general, higher concentrations were observed with the FlowCAM (although there were some exceptions in 2005). Relationships between physico-chemical and FlowCAM variables were studied and patterns of natural variability were identified. However, a clear response to nutrient pressure was not found. It is concluded that the potential of this automatic method for deriving cost-effective indicators is limited, especially in estuaries, due to the high interference with suspended material. Further studies incorporating fluorescence measurements with the FlowCAM may be more promising. The standardization of procedures is highly recommended.     

Characterization of Nontypable Haemophilus influenzae Isolates Recovered from Adult Patients with Underlying Chronic Lung Disease Reveals Genotypic and Phenotypic Traits Associated with Persistent Infection

Nontypable Haemophilus influenzae (NTHi) has emerged as an important opportunistic pathogen causing infection in adults suffering obstructive lung diseases. Existing evidence associates chronic infection by NTHi to the progression of the chronic respiratory disease, but specific features of NTHi associated with persistence have not been comprehensively addressed. To provide clues about adaptive strategies adopted by NTHi during persistent infection, we compared sequential persistent isolates with newly acquired isolates in sputa from six patients with chronic obstructive lung disease. Pulse field gel electrophoresis (PFGE) identified three patients with consecutive persistent strains and three with new strains. Phenotypic characterisation included infection of respiratory epithelial cells, bacterial self-aggregation, biofilm formation and resistance to antimicrobial peptides (AMP). Persistent isolates differed from new strains in showing low epithelial adhesion and inability to form biofilms when grown under continuous-flow culture conditions in microfermenters. Self-aggregation clustered the strains by patient, not by persistence. Increasing resistance to AMPs was observed for each series of persistent isolates; this was not associated with lipooligosaccharide decoration with phosphorylcholine or with lipid A acylation. Variation was further analyzed for the series of three persistent isolates recovered from patient 1. These isolates displayed comparable growth rate, natural transformation frequency and murine pulmonary infection. Genome sequencing of these three isolates revealed sequential acquisition of single-nucleotide variants in the AMP permease sapC, the heme acquisition systems hgpB, hgpC, hup and hxuC, the 3-deoxy-D-manno-octulosonic acid kinase kdkA, the long-chain fatty acid transporter ompP1, and the phosphoribosylamine glycine ligase purD. Collectively, we frame a range of pathogenic traits and a repertoire of genetic variants in the context of persistent infection by NTHi.     

Yeasts from Tierra Del Fuego Province (Argentina): Biodiversity,Characterization and Bioprospection of Hydrolytic Enzymes

Abstract

Antarctic and sub-Antarctic regions are – with Polar Regions, mountains and the deep sea – the most extreme environments on Earth because of its low temperatures, dryness, high incidence of solar radiation and low nutrient availability. Nevertheless, microorganisms have successfully colonized these regions. In this study, culturable yeasts from soil samples collected from two different locations, a human-impacted area (Encerrada Bay) and a largely pristine and naturally vegetated area near Lago Escondido city (54°39′0″S, 67°46′48″W) from Tierra del Fuego province, Argentina were identified and characterized at different levels. They were characterized and classified as psychrotolerant and were considered as moderately halotolerant because of their ability to grow in the presence of 1.5?M of NaCl. Yeasts from phylum Ascomycota were affiliated to five genera: Candida, Yarrowia, Debaryomyces, Nadsonia, and Wickerhamiella, whereas from phylum Basidiomycota yeasts were affiliated to six genera: Naganishia, Rhodotorula, Leucosporidum, Tausonia, Cystofilobasidium, and Apiotrichum. Most of the yeasts demonstrated at least one extracellular enzymatic activity (mainly β-glucosidase, esterase, and protease activities). One isolate identified as Tausonia pullulans showed significant activity across the eight enzyme types tested. In light of these findings, Tierra del Fuego province could be considered as a cold environment with a potential source of cold-adapted yeasts producing industrially relevant cold-active enzymes.     

Phytoplankton communities and biomass size structure (fractionated chlorophyll “a”), along trophic gradients of the Basque coast (northern Spain)

Eutrophication is a major threat to coastal ecosystems. Within Europe, the Water Framework Directive (WFD) has established the need of developing methods of assessment. Bulk chlorophyll “a” is used world-wide as an indicator of eutrophication. However, the size structure of the phytoplankton communities has not been investigated, in detail, in relation to eutrophication pressures. This study investigates the suitability of fractionated chlorophyll “a” (<10 and >10 μm), as an indicator of eutrophication. Along the Basque coast (northern Spain) several water bodies, ranging from offshore waters to the heads of the estuaries, were surveyed during 2008. Physico-chemical conditions and chlorophyll “a” showed a distinct spatial gradient. Trophic richness increased towards the middle and inner parts of the estuaries, where nutrient inputs, from natural or anthropogenic sources, together with the residence time of the water are usually higher. In summer, phytoplankton biomass and abundance decreased, generally, in offshore and coastal waters; in estuaries, they tended to increase. The chlorophyll distribution within the two size fractions was coherent with the phytoplankton taxonomic composition. In summer, the relative abundance of small-sized cells (diatoms and non-siliceous taxa) increased. The relationships between size-fractionated chlorophyll and physico-chemical variables differed, when comparing the offshore and coastal waters, with the estuaries. In the offshore and coastal waters, a strong seasonality was observed; in estuaries, an important spatial component was found. Fractionated chlorophyll provided complementary useful information on anthropogenic pressures. However, more research is necessary to utilise this variable as a tool for ecological status assessment, within the WFD.     

Elevated CO2 may impair the beneficial effect of arbuscular mycorrhizal fungi on the mineral and phytochemical quality of lettuce

Arbuscular mycorrhizal fungi (AMF) can improve growth and nutritional quality of greenhouse‐grown lettuces cultivated at ambient CO₂. Moreover, mycorrhizal symbiosis is predicted to be important in defining plant responses to elevated atmospheric CO₂ concentrations. Our main objective was to assess the effects of elevated CO₂ on growth and nutritional quality of greenhouse‐grown lettuces inoculated or not with AMF. Results showed that the accumulation of mineral nutrients (e.g. P, Cu, Fe) and antioxidant compounds (carotenoids, phenolics, anthocyanins, ascorbate) induced by AMF in leaves of lettuces cultivated at ambient CO₂ may diminish or disappear under elevated CO₂. It is hypothesized that a relevant quantity of photoassimilates could be used for improving shoot growth and spreading mycorrhizal colonization in detriment to the secondary metabolism. However, important differences can be found among different cultivars of lettuces.     

S-Ribosylhomocysteine analogues with the carbon-5 and sulfur atoms replaced by a vinyl or (fluoro)vinyl unit

Treatment of the protected ribose or xylose 5-aldehyde with sulfonyl-stabilized fluorophosphonate gave (fluoro)vinyl sulfones. Stannyldesulfonylation followed by iododestannylation afforded 5,6-dideoxy-6-fluoro-6-iodo-d-ribo or xylo-hex-5-enofuranoses. Coupling of the hexenofuranoses with alkylzinc bromides gave 10-carbon ribosyl- and xylosylhomocysteine analogues incorporating a fluoroalkene. The fluoroalkenyl and alkenyl analogues were evaluated for inhibition of Bacillus subtilis S-ribosylhomocysteinase (LuxS). One of the compounds, 3,5,6-trideoxy-6-fluoro-d-erythro-hex-5-enofuranose, acted as a competitive inhibitor of moderate potency (K_I = 96 μM).     

Testing mate choice and overdominance at MH in natural families of Atlantic salmon Salmo salar

This study aimed to test mate choice and selection during early life stages on major histocompatibility (MH) genotype in natural families of Atlantic salmon Salmo salar spawners and juveniles, using nine microsatellites to reconstruct families, one microsatellite linked to an MH class I gene and one minisatellite linked to an MH class II gene. MH‐based mate choice was only detected for the class I locus on the first year, with lower expected heterozygosity in the offspring of actually mated pairs than predicted under random mating. The genotype frequencies of MH‐linked loci observed in the juveniles were compared with frequencies expected from Mendelian inheritance of parental alleles to detect selection during early life stages. No selection was detected on the locus linked to class I gene. For the locus linked to class II gene, observed heterozygosity was higher than expected in the first year and lower in the second year, suggesting overdominance and underdominance, respectively. Within family, juveniles' body size was linked to heterozygosity at the same locus, with longer heterozygotes in the first year and longer homozygotes in the second year. Selection therefore seems to differ from one locus to the other and from year to year.     

Biometeorological comfort index

The paper aims to represent and scale biometeorological sensations through a thermal index which comprises the effects of temperature, humidity and wind. The cooling power of a measured combination of wind speed and air temperature is equated with that due to the combination of a reference wind speed (related to the air movement experienced by a person walking in still air), and a derived aerodynamic temperature. A biometeorological temperature which is the average of such an aerodynamic temperature and a conventionally measured wet bulb temperature is shown to afford a better measure of comfort in Spain than some other, alternative indices.