Functional evidence for the inflammatory reflex in teleosts: A novel alpha7 nicotinic acetylcholine receptor modulates the macrophage response to dsRNA

The inflammatory reflex modulates the innate immune system, keeping in check the detrimental consequences of overstimulation. A key player controlling the inflammatory reflex is the alpha 7 acetylcholine receptor (α7nAChR). This receptor is one of the signalling molecules regulating cytokine expression in macrophages. In this study, we characterize a novel teleost α7nAChR. Protein sequence analysis shows a high degree of conservation with mammalian orthologs and trout α7nAChR has all the features and essential amino acids to form a fully functional receptor. We demonstrate that trout macrophages can bind α-bungarotoxin (α-BTX), a competitive antagonist for α7nAChRs. Moreover, nicotine stimulation produces a decrease in pro-inflammatory cytokine expression after stimulation with poly(I:C). These results suggest the presence of a functional α7nAChR in the macrophage plasma membrane. Further, in vivo injection of poly(I:C) induced an increase in serum ACh levels in rainbow trout. Our results manifest for the first time the functional conservation of the inflammatory reflex in teleosts

Comparative analysis of the acute response of the trout, O. mykiss, head kidney to in vivo challenge with virulent and attenuated infectious hematopoietic necrosis virus and LPS-induced inflammation

Background: The response of the trout, O. mykiss, head kidney to bacterial lipopolysaccharide (LPS) or active and attenuated infectious hematopoietic necrosis virus (IHNV and attINHV respectively) intraperitoneal challenge, 24 and 72 hours post-injection, was investigated using a salmonid-specific cDNA microarray. Results: The head kidney response to i.p. LPS-induced inflammation in the first instance displays an initial stress reaction involving suppression of major cellular processes, including immune function, followed by a proliferative hematopoietic-type/biogenesis response 3 days after administration. The viral response at the early stage of infection highlights a suppression of hematopoietic and protein biosynthetic function and a stimulation of immune response. In fish infected with IHNV a loss of cellular function including signal transduction, cell cycle and transcriptional activity 72 hours after infection reflects the tissue-specific pathology of IHNV infection. attIHNV treatment on the other hand shows a similar pattern to native IHNV infection at 24 hours however at 72 hours a divergence from the viral response is seen and replace with a recovery response more similar to that observed for LPS is observed. Conclusion: In conclusion we have been able to identify and characterise by transcriptomic analysis two different types of responses to two distinct immune agents, a virus, IHNV and a bacterial cell wall component, LPS and a 'mixed' response to an attenuated IHNV. This type of analysis will lead to a greater understanding of the physiological response and the development of effective immune responses in salmonid fish to different pathogenic and pro-inflammatory agents

A reference architecture for cloud-edge meta-operating systems enabling cross-domain, data-intensive, ML-assisted applications: architectural overview and key concepts

Future data-intensive intelligent applications are required to traverse across the cloudto-edge-to-IoT continuum, where cloud and edge resources elegantly coordinate, alongside sensor networks and data. However, current technical solutions can only partially handle the data outburst associated with the IoT proliferation experienced in recent years, mainly due to their hierarchical architectures. In this context, this paper presents a reference architecture of a meta-operating system (RAMOS), targeted to enable a dynamic, distributed and trusted continuum which will be capable of facilitating the next-generation smart applications at the edge. RAMOS is domain-agnostic, capable of supporting heterogeneous devices in various network environments. Furthermore, the proposed architecture possesses the ability to place the data at the origin in a secure and trusted manner. Based on a layered structure, the building blocks of RAMOS are thoroughly described, and the interconnection and coordination between them is fully presented. Furthermore, illustration of how the proposed reference architecture and its characteristics could fit in potential key industrial and societal applications, which in the future will require more power at the edge, is provided in five practical scenarios, focusing on the distributed intelligence and privacy preservation principles promoted by RAMOS, as well as the concept of environmental footprint minimization. Finally, the business potential of an open edge ecosystem and the societal impacts of climate net neutrality are also illustrated.For UPC authors: this research was funded by the Spanish Ministry of Science, Innovation and Universities and FEDER, grant number PID2021-124463OB-100.Peer ReviewedPostprint (published version

RNA-Seq Reveals an Integrated Immune Response in Nucleated Erythrocytes

Background: Throughout the primary literature and within textbooks, the erythrocyte has been tacitly accepted to have maintained a unique physiological role; namely gas transport and exchange. In non-mammalian vertebrates, nucleated erythrocytes are present in circulation throughout the life cycle and a fragmented series of observations in mammals support a potential role in non-respiratory biological processes. We hypothesised that nucleated erythrocytes could actively participate via ligand-induced transcriptional re-programming in the immune response. Methodology/Principal Findings: Nucleated erythrocytes from both fish and birds express and regulate specific pattern recognition receptor (PRR) mRNAs and, thus, are capable of specific pathogen associated molecular pattern (PAMP) detection that is central to the innate immune response. In vitro challenge with diverse PAMPs led to de novo specific mRNA synthesis of both receptors and response factors including interferon-alpha (IFNα) that exhibit a stimulus-specific polysomal shift supporting active translation. RNA-Seq analysis of the PAMP (Poly (I:C), polyinosinic:polycytidylic acid)-erythrocyte response uncovered diverse cohorts of differentially expressed mRNA transcripts related to multiple physiological systems including the endocrine, reproductive and immune. Moreover, erythrocyte-derived conditioned mediums induced a type-1 interferon response in macrophages thus supporting an integrative role for the erythrocytes in the immune response. Conclusions/Significance: We demonstrate that nucleated erythrocytes in non-mammalian vertebrates spanning significant phylogenetic distance participate in the immune response. RNA-Seq studies highlight a mRNA repertoire that suggests a previously unrecognized integrative role for the erythrocytes in other physiological systems

Characterization and expression of NADPH oxidase in LPS-, poly(I:C)- and zymosan-stimulated trout (Oncorhynchus mykiss W.) macrophages

In vertebrates, the generation of superoxide reactive oxygen species (ROS) via activation of the Nox/Duox family of NADPH oxidases is a prototypical feature of the pathogen-induced defensive responses of activated professional phagocytes. To understand the role of the rainbow trout (Oncorhynchus mykiss) Phox oxidase from a phylogenetic and functional perspective we describe the cloning, sequencing and expression analysis of multiple NADPH components in cultured macrophages. Phylogenetic analyses support the notion of the emergence of Phox-related components before the diversification of basal euteleosts and add to the limited collection of teleost NADPH oxidases. Expression studies using lipopolysaccharide, polyinosine-polycytidylic acid and zymosan to mimic the onset of inflammatory responses in trout macrophages suggest differences in regulation of the NADPH complex throughout the maturation/differentiation period of culture and between different treatments

Molecular cloning and characterization of European seabass (Dicentrarchus labrax) and Gilthead seabream (Sparus aurata) complement component C3

13 páginas, 6 figuras, 1 tabla, 1 apéndiceWe present the complete C3 cDNA sequence of Gilthead seabream (Sparus aurata) and European seabass (Dicentrarchus labrax) and its molecular characterization with a descriptive analysis of their structural elements. We obtained one sequence for Gilthead seabream (gsbC3) which encodes a predicted protein of 1656 amino acids, and two sequences for European seabass (esbC3_1 and esbC3_2) which encode two predicted proteins of 1654 and 1587 amino acids respectively. All sequences present the characteristic structural features of C3 but interestingly esbC3_2 lacks the anaphylotoxin domain and the cysteine residue responsible for thiolester bond formation. Moreover, we have detected and quantified (by realtime PCR-based absolute quantification) specific isoform expression in European seabass depending on pathogen and density conditions in vivo. In addition, we have analyzed the tissue distribution pattern of European seabass and Gilthead seabream C3 genes under crowding stress and under pathological challenges in vivo, and we have observed that crowding and infection status provoke changes in expression levels, tissue expression pattern and C3 isoform expression balance.This work has been supported by the Imaquanim project (Immunity of Aquatic Animals. UE 6th framework program) and BFU2009-07354 (Ministry of Science and Innovations) and SGR2009- 554 (Generalitat de Catalunya).Peer reviewe

Low-Frequency Atmospheric Variability Patterns and Synoptic Types Linked to Large Floods in the Lower Ebro River Basin

This study analyzes the atmospheric variability that caused the largest floods affecting the town of Tortosa, Spain, in the mouth of the Ebro River (northeast Iberian Peninsula). The Tortosa flood database and flood marks in the nearby town of Xerta are used to define the more relevant flooding episodes (discharges.2900 m s) of the 1600-2005 period. We explore the atmospheric variability based on low-frequency patterns and synoptic types applying a multivariable analysis to grids at sea level pressure and geopotential at 500 hPa provided by the twentieth-century V3 Reanalysis Project for the instrumental period (since 1836). Output from the Last Millennium Ensemble Project was used to analyze the sea level pressure over the pre-instrumental period (before 1836). Our analysis includes 33 flood episodes. Four synoptic types are related to floods in Tortosa since 1836, characterized by low pressure systems that interact with the Mediterranean warm air mass and promote atmospheric destabilization. Flooding in Tortosa is related to relative high values of solar activity, positive Northern Hemisphere temperature anomalies, and NAO in positive phase. This result indicates that the major floods are related to zonal atmospheric circulations (west-to-east cyclone transfer). During winter, the main impact of the floods is located at the western part of the basin, and the Pyrenean subbasins are affected during autumn. The major finding is that similar flood behavior is detected since 1600, improving our understanding of past climates, enhancing the knowledge base for some aspects and impacts of climate change, and reducing uncertainty about future outcomes. SIGNIFICANCE STATEMENT: A total of 33 large floods (.2900 m s) were registered since 1600 in Tortosa, Spain, located at the mouth of the Ebro River (northeast Iberian Peninsula). They occur associated with low pressure systems that interact with the Mediterranean warm air mass promoting atmospheric destabilization. The floods in Tortosa are also associated with other important processes occurring at significantly longer time scales: high values of solar activity, positive Northern Hemisphere temperature anomalies, and NAO in positive phase, indicating that the major floods are related to zonal atmospheric circulations. The major finding is that we detect similar flood behaviors since 1600, improving our understanding of past climates, enhancing the knowledge base for some aspects and impacts of climate change, and reducing uncertainty about future outcomes

Comparative analysis of the acute response of the trout, , head kidney to challenge with virulent and attenuated infectious hematopoietic necrosis virus and LPS-induced inflammation-4

Ata is shown as mean fold change ± std.error.<p><b>Copyright information:</b></p><p>Taken from "Comparative analysis of the acute response of the trout, , head kidney to challenge with virulent and attenuated infectious hematopoietic necrosis virus and LPS-induced inflammation"</p><p>http://www.biomedcentral.com/1471-2164/9/141</p><p>BMC Genomics 2008;9():141-141.</p><p>Published online 26 Mar 2008</p><p>PMCID:PMC2291046.</p><p></p