Evolution of Th2 responses : Characterization of IL-4/13 in sea bass (Dicentrarchus labrax L.) and studies of expression and biological activity

Acknowledgements This research was funded by the European Commission under the 7th Framework Programme for Research and Technological Development (FP7) of the European Union (Grant Agreement 311993 TARGETFISH). T.W. received funding from the MASTS pooling initiative (The Marine Alliance for Science and Technology for Scotland). MASTS is funded by the Scottish Funding Council (grant reference number HR09011) and contributing institutions.Peer reviewedPublisher PD

Controls on natural gas migration in the western Nile Delta fan

The aim of this study is to combine petrophysical and geochemical data in order to reconstruct the migration history and pathways of mixed microbial-thermogenic gases drilled on the Nile Delta fan, offshore Egypt. While general interest lies in understanding migration routes, rates and mechanisms special attention is dedicated to understanding (1) the origin of gas in both reservoir and non-reservoir units using chemical and isotopic fingerprints and (2) whether a free gas phase supports relatively rapid leakage via bulk flow in non-reservoir units, both above and below commercial accumulations. The Pilocene section in this study is a classic slope environment comprising channels, mud-rich turbidites, mass transport complexes and hemipelagites. Data from seismic and drilled wells suggest that the channel and levee reservoirs are rarely full to spill, implying either a lack of charge and leakage rates which precludes complete filling of the structures. The provided data set enables a quantitative assessment of gas distribution and its genetic fingerprint in the context of both stratigraphic position and lithology. Data is reported from 25 wells, each provided with a conventional wireline log suite and some with borehole images and high-quality core images. Gas concentration data, plus compositional and isotope data are available for isotubes and headspace gas for both reservoir and non-reservoir units. Small-to-medium scale linear and non-linear depth shifts between different techniques (core recovery, core logging, wireline logging) in conjunction with scale and resolution issues demanded logical/stochastic depth synchronisation and well as harmonisation of signal resolution (typically up-scaling). Accordingly, great care was taken to depth-match core, log and gas data. In general, there is evidence of leaking thermogenic and partly biodegraded gas from the reservoirs under investigation, while some microbial methane appears to be retained in the cap rock. Careful assessment of the maturity of the thermogenic gas charge suggests that in a given structure, maturities are similar throughout the sampled section of underseal, reservoir and top seal. Furthermore, compositional temperature stratification suggests a balance between influx of fresh gas and microbial metabolism rates, supporting the concept of a dynamic charge-leak scenario that is sustaining hydrocarbon fermenting microbial communities in the deep biosphere. It was found that microbial recycling of hydrocarbons at depth enables the identification of diffusive gas mixing pathways in the combined analysis of methane and ethane compositional and isotopic data. The proposed diffusion pattern supports the idea of a widely present coupling between both methanogenic and biodegrading microbial communities that exhibit strong carbon isotopic dis-balances at gas-water contacts (GWC) where nutrient supply is in favour of the biodegrading metabolism. Although the hypothesis of coupled diffusive/microbial gas overprints complies with (1) various literature reports that microbial attack on free gas phases is hindered by restricted physical access and (2) segregative isotope fractionation as a consequence of differences on methane and ethane diffusivity, it is conditional to the nature of gas mixing patterns along borehole trajectories in the context of lithology and pore fluid saturations. Undoubtedly, the ubiquitous presence of microbial gas has consequences for vertical net leakage. As classic empirical wireline models for hydrocarbon saturation (i.e. free gas phase volumetrics) are not suited for clay-dominated cap rock sections, an alternative approach presented in this study is based on total gas (TG) modelling from nuclear logs and its solubility in the formation of brine. The calibrated saturation model is scale-independent and implies that free gas occurs on the most of the clay-dominated non-reservoir sections. However, model resolution is not sufficient to capture the suspected filamentary network of free gas phase within the mudrock pore space that enables relatively rapid leakage via Darcy flow. In an unique attempt to validate manual and thereby subjective lithofacies allocations to core images a subset of rock sample properties such as grain size fractions and porosity were successfully modelled using quantitative core image properties. However, model validity appears to be restricted to clay-rich lithofacies due to both an absence of calibration data for sands and occurrence of abnormally dark sandstone units. Further, an artificial neural network (ANN) was trained to propagate the calibrated core fancies along the entire wireline logged borehole section to set the lithological context for a detailed fluid flow analysis. Reproducibility of input (core) facies by output (wireline) facies is similar to the reproducibility by fellow geoscientists but could not be significantly improved to 60-80% of reliability by reduction of facies types. The study shows that a combination of geochemical data with lithological and petro-physical information generates detailed insights into rates, mechanisms, and pathways of two phase flow through the deep biosphere of gas-charged basins. Vertical, geologically rapid flow through mud-rich sequences is a viable migration route for gas if the influence of cap rock bypass systems (permeable faults, sandstone intrusions, mud volcanoes etc.) is restricted. It was found that an adequate quantification of both thermogenic gas fraction and diffusive gas mixing fingerprints is crucial to identity stratigraphic intervals that are not dominated by advective leakage through the pore space and are consequently bypassed.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

TGF-β1 Exerts Opposing Effects on Grass Carp Leukocytes: Implication in Teleost Immunity, Receptor Signaling and Potential Self-Regulatory Mechanisms

In fish immunity, the regulatory role of transforming growth factor-β1 (TGF-β1) has not been fully characterized. Here we examined the immunoregulatory effects of TGF-β1 in grass carp peripheral blood leukocytes (PBL) and head kidney leukocytes (HKL). It is interesting that TGF-β1 consistently stimulated the cell viability and the mRNA levels of pro-inflammatory cytokines (Tnfα and Ifnγ) and T/B cell markers [Cd4-like (Cd4l), Cd8α, Cd8β and Igμ] in PBL, which contrasted with its inhibitory tone in HKL. Further studies showed that grass carp TGF-β1 type I receptor, activin receptor-like kinase 5 (ALK5), was indispensable for the immunoregulatory effects of TGF-β1 in PBL and HKL. Notably, TGF-β1 persistently attenuated ALK5 expression, whereas immunoneutralization of endogenous grass carp TGF-β1 could increase ALK5 mRNA and protein levels. It is consistent with the observation that TGF-β1 decreased the number of ALK5+ leukocytes in PBL and HKL, revealing a negative regulation of TGF-β1 signaling at the receptor level. Moreover, transient treatment with TGF-β1 for 24 h was sufficient to induce similar cellular responses compared with the continuous treatment. This indicated a possible mechanism by which TGF-β1 triggered the down-regulation of ALK5 mRNA and protein, leading to the desensitization of grass carp leukocytes toward TGF-β1. Accordingly, our data revealed a dual role of TGF-β1 in teleost immunity in which it can serve as a positive or negative control device and provided additional mechanistic insights as to how TGF-β1 controls its signaling in vertebrate leukocytes

An Enriched European Eel Transcriptome Sheds Light upon Host-Pathogen Interactions with Vibrio vulnificus

Infectious diseases are one of the principal bottlenecks for the European eel recovery. The aim of this study was to develop a new molecular tool to be used in host-pathogen interaction experiments in the eel. To this end, we first stimulated adult eels with different pathogen-associated molecular patterns (PAMPs), extracted RNA from the immune-related tissues and sequenced the transcriptome. We obtained more than 2 x 10(6) reads that were assembled and annotated into 45,067 new descriptions with a notable representation of novel transcripts related with pathogen recognition, signal transduction and the immune response. Then, we designed a DNA-microarray that was used to analyze the early immune response against Vibrio vulnificus, a septicemic pathogen that uses the gills as the portal of entry into the blood, as well as the role of the main toxin of this species (RtxA13) on this early interaction. The gill transcriptomic profiles obtained after bath infecting eels with the wild type strain or with a mutant deficient in rtxA13 were analyzed and compared. Results demonstrate that eels react rapidly and locally against the pathogen and that this immune-response is rtxA13-dependent as transcripts related with cell destruction were highly up-regulated only in the gills from eels infected with the wild-type strain. Furthermore, significant differences in the immune response against the wild type and the mutant strain also suggest that host survival after V. vulnificus infection could depend on an efficient local phagocytic activity. Finally, we also found evidence of the presence of an interbranchial lymphoid tissue in European eel gills although further experiments will be necessary to identify such tissue

Identification of Novel Proteins in Neospora caninum Using an Organelle Purification and Monoclonal Antibody Approach

Neospora caninum is an important veterinary pathogen that causes abortion in cattle and neuromuscular disease in dogs. Neospora has also generated substantial interest because it is an extremely close relative of the human pathogen Toxoplasma gondii, yet does not appear to infect humans. While for Toxoplasma there are a wide array of molecular tools and reagents available for experimental investigation, relatively few reagents exist for Neospora. To investigate the unique biological features of this parasite and exploit the recent sequencing of its genome, we have used an organelle isolation and monoclonal antibody approach to identify novel organellar proteins and develop a wide array of probes for subcellular localization. We raised a panel of forty-six monoclonal antibodies that detect proteins from the rhoptries, micronemes, dense granules, inner membrane complex, apicoplast, mitochondrion and parasite surface. A subset of the proteins was identified by immunoprecipitation and mass spectrometry and reveal that we have identified and localized many of the key proteins involved in invasion and host interaction in Neospora. In addition, we identified novel secretory proteins not previously studied in any apicomplexan parasite. Thus, this organellar monoclonal antibody approach not only greatly enhances the tools available for Neospora cell biology, but also identifies novel components of the unique biological characteristics of this important veterinary pathogen

Characterisation and expression analysis of the Atlantic halibut (Hippoglossus hippoglossus L.) cytokines: IL-1β, IL-6, IL-11, IL-12β and IFNγ

Genes encoding the five Atlantic halibut (Hippoglossus hippoglossus L.) cytokines; interleukin (IL)-1β, IL-6, IL-11b, IL-12βc, and interferon (IFN) γ, were cloned and characterised at a molecular level. The genomic organisation of the halibut cytokine genes was similar to that seen in mammals and/or other fish species. Several mRNA instability motifs were found within the 3′-untranslated region (UTR) of all cytokine cDNA sequences. The putative cytokine protein sequences showed a low sequence identity with the corresponding homologues in mammals, avian and other fish species. Nevertheless, important structural features were presumably conserved such as the presence, or absence in the case of IL-1β, of a signal peptide, secondary structure and family signature motifs. The relative expression pattern of the cytokine genes was analyzed in several halibut organs, revealing a constitutive expression in both lymphoid and non-lymphoid organs. Interestingly, the gills showed a relatively high expression of IL-1β, IL-12βc and IFNγ. The real time RT-PCR data also showed that the mRNA level of IL-1β, IL-6, IL-12βc and IFNγ was high in the thymus, while IL-11b was relatively highly expressed in the posterior kidney and posterior gut. Moreover, the halibut brain showed a relatively high level of IL-6 transcripts. Anterior kidney leucocytes in vitro stimulated with imiquimod showed a significant increase in mRNA level of the five halibut cytokine genes. The sequence and characterisation data presented here will be useful for further investigation of both innate and adaptive immune responses in halibut, and be helpful in the design of vaccines for the control of various infectious diseases

Education, implementation, and teams : 2020 International Consensus on Cardiopulmonary Resuscitation and Emergency Cardiovascular Care Science with treatment recommendations

For this 2020 International Consensus on Cardiopulmonary Resuscitation and Emergency Cardiovascular Care Science With Treatment Recommendations, the Education, Implementation, and Teams Task Force applied the population, intervention, comparator, outcome, study design, time frame format and performed 15 systematic reviews, applying the Grading of Recommendations, Assessment, Development, and Evaluation guidance. Furthermore, 4 scoping reviews and 7 evidence updates assessed any new evidence to determine if a change in any existing treatment recommendation was required. The topics covered included training for the treatment of opioid overdose; basic life support, including automated external defibrillator training; measuring implementation and performance in communities, and cardiac arrest centers; advanced life support training, including team and leadership training and rapid response teams; measuring cardiopulmonary resuscitation performance, feedback devices, and debriefing; and the use of social media to improve cardiopulmonary resuscitation application

Structure-function relationships of pheromones of the ciliate Euplotes raikovi with mammalian growth factors: cross-reactivity between Er-1 and interleukin-2 systems.

Diffusible protein signals of the ciliate Euplotes raikovi, denoted as pheromones, have functionally been linked with prototypic growth factors of animal cells by the demonstration that they not only induce a temporary cell union in mating pairs, by acting in a paracrine-like fashion, but can also bind to cells in autocrine fashion and promote their vegetative (mitotic) proliferation. It is now shown that pheromone Er-1 is capable of binding to the alpha and beta chains of the multimeric IL-2 receptor on mammalian cells and that IL-2 can, in turn, bind to the putative cell receptor of this pheromone. Similarities in the IL-2 and Er-1 structures support these findings and raise controversial implications with regard to their evolutionary significance

Fish immunology after the post-genomic era.

The research on fish immunology has risen impressively over the last decades, providing deep insights into the evolution of the immune system of vertebrates and underlying genetic networks, but also discovering some peculiarities, mainly related to ancestral or heavily specialised molecular and cellular characters. Gathered information has built a sound platform to further accomplish the elucidation of evolutionary processes and boost application-based research to monitor fish health and contrast old and emerging diseases. The extensive cloning of genes encoding for immune-regulatory molecules, together with the sequencing of many fish genomes, provided scientists with a comprehensive panel of nucleotide sequences. Therefore, fish have become the main research models for cold-blooded vertebrates. It is expected in the near future that much research will be addressed to revive the more arduous work remaining, namely, that of associating molecules, cell features and anatomy with function. Considerable efforts have been, and will be, directed toward a careful interpretation of differences among species, disclosure of relation to fish habits, and interactions of individual specimens with the changing (and challenging) environment. When proposing expert reviews on some research topics that pursue these intriguing perspectives, this Special Issue of Fish and Shellfish Immunology aimed at updating current knowledge on fish immunology. This Special Issue arises from outstanding contributions presented at the Symposium of the European Organisation of Fish Immunology (EOFFI) held in Viterbo, Italy, on May 2010. As a result of the wide and qualified participation of scientists from all the continents, the Organisation has been renamed as the new International Society of Fish and Shellfish Immunology (ISFSI, www.isfsim.org), aiming at extending the scientific interests and activities to shellfish immunology