Zebrafish fin immune responses during high mortality infections with viral haemorrhagic septicemia rhabdovirus. A proteomic and transcriptomic approach

<p>Abstract</p> <p>Background</p> <p>Despite rhabdoviral infections being one of the best known fish diseases, the gene expression changes induced at the surface tissues after the natural route of infection (infection-by-immersion) have not been described yet. This work describes the differential infected versus non-infected expression of proteins and immune-related transcripts in fins and organs of zebrafish <it>Danio rerio </it>shortly after infection-by-immersion with viral haemorrhagic septicemia virus (VHSV).</p> <p>Results</p> <p>Two-dimensional differential gel electrophoresis detected variations on the protein levels of the enzymes of the glycolytic pathway and cytoskeleton components but it detected very few immune-related proteins. Differential expression of immune-related gene transcripts estimated by quantitative polymerase chain reaction arrays and hybridization to oligo microarrays showed that while more transcripts increased in fins than in organs (spleen, head kidney and liver), more transcripts decreased in organs than in fins. Increased differential transcript levels in fins detected by both arrays corresponded to previously described infection-related genes such as complement components (<it>c3b, c8 </it>and <it>c9</it>) or class I histocompatibility antigens (<it>mhc1</it>) and to newly described genes such as secreted immunoglobulin domain (<it>sid4</it>), macrophage stimulating factor (<it>mst1</it>) and a cluster differentiation antigen (<it>cd36</it>).</p> <p>Conclusions</p> <p>The genes described would contribute to the knowledge of the earliest molecular events occurring in the fish surfaces at the beginning of natural rhabdoviral infections and/or might be new candidates to be tested as adjuvants for fish vaccines.</p

Rainbow trout surviving infections of viral haemorrhagic septicemia virus (vhsv) show lasting antibodies to recombinant g protein fragments

P. 929-935Rainbow trout antibodies (Abs) binding to recombinant fragments (frgs) derived from the protein G of the viral haemorrhagic septicemia virus (VHSV)-07.71 strain, could be detected by ELISA (frg-ELISA) in sera from trout surviving laboratory-controlled infections. Abs were detected not only by using sera from trout infected with the homologous VHSV isolate but also with the VHSV-DK-201433 heterologous isolate, which had 13 amino acid changes. Sera from healthy trout and/or from trout surviving infectious haematopoietic necrosis virus (IHNV) infection, were used to calculate cut-off absorbances to differentiate negative from positive sera. Specific anti-VHSV Abs could then be detected by using any of the following frgs: frg11 (56-110), frg15 (65-250), frg16 (252-450) or G21-465. While high correlations were found among the ELISA values obtained with the different frgs, no correlations between any frg- ELISA and complement-dependent 50 % plaque neutralization test (PNT) titres could be demonstrated. Between 4 to 10 weeks after VHSV-infection, more trout sera were detected as positives by using heterologous frg-ELISA rather than homologous PNT. Furthermore, the percentage of positive sera detected by frg11-ELISA increased with time after infection to reach 100 %, while those detected by complement-dependent PNT decreased to 29.4 %, thus confirming that the lack of neutralising Abs does not mean the lack of any anti-VHSV Abs in survivor trout sera. Preliminary results with sera from field samples suggest that further refinements of the frg-ELISA could allow detection of anti-VHSV trout Abs in natural outbreaks caused by different heterologous VHSV isolates.The homologous frg-ELISA method could be useful to follow G immunization attempts during vaccine development and/or to best understand the fish Ab response during VHSV infections. The viral frgs approach might also be used with other fish species and/or viruses.S

Effect of the filler on the nanomechanical properties of polypropylene in contact with paraffinic phase change material

As the changes on the mechanical properties with depth of indentation provide valuable information that may be suitable for design purposes, nanoindentation is an adequate technique for investigating the nanomechanical changes in the surface and in the inner polymers. This research focuses on the study of the nanomechanical properties of two grades of polypropylene (PP) after the long term exposure to an organic fluid, such as paraffin wax used as a phase change materials (PCM). PCM are used for thermal energy storage (TES) in buildings applications for passive systems or heating and cooling usages. In any of these uses PCM must be encapsulated and PP is a possibility within the container materials used. Four different samples of the polypropylene were evaluated: PP, filled polypropylene with 60% Mg(OH)2 (PP-60Mg), PP-60Mg with PCM (RT-25), and PP-60Mg with PCM (RT-42). It was studied the thermal stability by Thermogravimetrical analysis of these samples, and also it was evaluated the Hardness (H), Elastic modulus (E) and Loss modulus (Eloss) for the unfilled PP and PP-60Mg in contact with two different PCM at different temperatures (30 °C, 45 °C and 60 °C for RT-25 and 45 °C for RT-42) for 32 days. Results show that the mechanical properties H, E, and Eloss are higher for PP-60Mg than for PP. Nevertheless, these properties decrease significantly when the PP and the PP-60Mg are in contact with PCM, because it acts as a plasticizer, softening the polymer. Besides, the higher PCM's melting point, the lowest mechanical properties were observed

Depth-sensing indentation applied to polymers: a comparison between standard methods of analysis in relation to the nature of the materials

Mechanical data (hardness and elastic modulus) from instrumented indentation testing are often extracted assuming linear elasticity in the initial portion of the unloading. The method is nowadays widely accepted as a convenient tool to interpret depth-sensing data, however it is a matter of controversy when applied to polymer materials due to their time-dependent behaviour. More recently, Loubet and co-workers applied continuous stiffness measurements (CSM), consisting of superimposing a small oscillation to the quasi-static component of loading, to the study of the mechanical properties of polymers and proposed a new model to account for the apparent increase in the contact area detected at the first stages of contact. The present work offers a comparative study between the Loubet‟s model using CSM and the procedure yielding a single reading from the onset of unloading. A wide range of thermoplastic polymer materials including glassy and semicrystalline polymers have been investigated. The most important equations employed for each method are summarized and the advantages and disadvantages of employing one procedure or the other are discussed. The differences found between the results obtained from both approaches are discussed in relation to the nature of the polymer material. A comparison between mechanical data extracted from indentation measurements and from classical dynamic mechanical analysis is offered

Antibody recognition of the glycoprotein g of viral haemorrhagic septicemia virus (VHSV) purified in large amounts from insect larvae

<p>Abstract</p> <p>Background</p> <p>There are currently no purification methods capable of producing the large amounts of fish rhabdoviral glycoprotein G (gpG) required for diagnosis and immunisation purposes or for studying structure and molecular mechanisms of action of this molecule (ie. pH-dependent membrane fusion). As a result of the unavailability of large amounts of the gpG from viral haemorrhagic septicaemia rhabdovirus (VHSV), one of the most dangerous viruses affecting cultured salmonid species, research interests in this field are severely hampered. Previous purification methods to obtain recombinant gpG from VHSV in <it>E. coli</it>, yeast and baculovirus grown in insect cells have not produced soluble conformations or acceptable yields. The development of large-scale purification methods for gpGs will also further research into other fish rhabdoviruses, such as infectious haematopoietic necrosis virus (IHNV), spring carp viremia virus (SVCV), hirame rhabdovirus (HIRRV) and snakehead rhabdovirus (SHRV).</p> <p>Findings</p> <p>Here we designed a method to produce milligram amounts of soluble VHSV gpG. Only the transmembrane and carboxy terminal-deleted (amino acid 21 to 465) gpG was efficiently expressed in insect larvae. Recognition of G21-465 by ß-mercaptoethanol-dependent neutralizing monoclonal antibodies (N-MAbs) and pH-dependent recognition by sera from VHSV-hyperimmunized or VHSV-infected rainbow trout (<it>Oncorhynchus mykiss</it>) was demonstrated.</p> <p>Conclusions</p> <p>Given that the purified G21-465 conserved some of its most important properties, this method might be suitable for the large-scale production of fish rhabdoviral gpGs for use in diagnosis, fusion and antigenicity studies.</p

Nuevas terapias dirigidas para el tratamiento del cáncer

El cáncer es el término que se utiliza para englobar un conjunto de enfermedades que se caracterizan por el crecimiento descontrolado de células alteradas molecularmente por mutaciones o modificaciones epigenéticas.En la presente revisión describimos algunas terapias dirigidas que se están utilizando actualmente en clínic

Study of the future trend of road freight transport

The road freight transport is facing one of the biggest changes in history. This turning point is motivated by a cultural revolution, a new vision for the environment, a higher concern for road safety and a need to preserve the earth and its resources. The road freight market as we know it has been dominated by diesel trucks for over a century. During all this time, the amount of CO2 produced and emitted by light and heavy-duty vehicles is practically incalculable. The road transportation industry is the principal pollutant and emitter of CO2 of all sectors, which is why there must be an end to the fossil fuel era. But how will this be possible to happen in the near future? The main manufacturers around the world have been investigating and developing plenty of solutions during the last decade. The aim of this project is to study some of the most developed future technologies that might be implemented in road freight vehicles, including light-duty vehicles, medium-freight trucks and heavy-freight trucks. Even though most of the technologies and developments will be seen in most of the world’s markets, this project focuses mainly on the study of the European market. The first part of the report is destined to study the actual market, its necessities and the present and future challenges such as environmental effects, CO2 emissions, road safety or resource depletion. The second part is focused on studying the main four technologies that will most likely be seen in a nearby future in the road freight transport sector. Those technologies are biofuels, to replace fossil fuels; hybrid and electric trucks; hydrogen and fuel cells powered trucks; and autonomous trucks. The research includes a wide technological, economical, and sustainable study for each proposal

Ambient temperature modulates body weight changes in patients with advanced oncological diseases and anorexia cachexia syndrome

A prospective naturalistic multicenter study of patients undergoing oncological treatment at four hospitals during a three-year period (2017–2020) in the Autonomous Community of Extremadura in southwestern Spain with a continentalized Mediterranean climate of mild and relatively rainy winters, and particularly hot and sunny summers. Bodyweight changes were obtained from the medical records of 84 oncological patients (59 men and 25 women, age range 37–91 yrs). Mean monthly AT was used to examine the association of weight changes across cold and warm bimesters -BIMs (December and January, vs. July and August), Trimesters -TRIMs (July to September vs. December to February), and Semesters -SEMs (May to October vs. November to April). Weight changes between two consecutive weight measures were categorized as weight gain, weight loss, or no weight change. Differences across cold and warm seasons were analysed using parametric (ANOVA), and nonparametric statistics (Chi-square and binomial z tests). An alpha-rate of 0.05 was used for all analysesS

Differential Immune Transcriptome and Modulated Signalling Pathways in Rainbow Trout Infected with Viral Haemorrhagic Septicaemia Virus (VHSV) and Its Derivative Non-Virion (NV) Gene Deleted

17 Pág.Viral haemorrhagic septicaemia virus (VHSV) is one of the worst viral threats to fish farming. Non-virion (NV) gene-deleted VHSV (dNV-VHSV) has been postulated as an attenuated virus, because the absence of the NV gene leads to lower induced pathogenicity. However, little is known about the immune responses driven by dNV-VHSV and the wild-type (wt)-VHSV in the context of infection. Here, we obtained the immune transcriptome profiling in trout infected with dNV-VHSV and wt-VHSV and the pathways involved in immune responses. As general results, dNV-VHSV upregulated more trout immune genes than wt-VHSV (65.6% vs 45.7%, respectively), whereas wt-VHSV maintained more non-regulated genes than dNV-VHSV (45.7% vs 14.6%, respectively). The modulated pathways analysis (Gene-Set Enrichment Analysis, GSEA) showed that, when compared to wt-VHSV infected trout, the dNV-VHSV infected trout upregulated signalling pathways (n = 19) such as RIG-I (retinoic acid-inducible gene-I) like receptor signalling, Toll-like receptor signalling, type II interferon signalling, and nuclear factor kappa B (NF-kappa B) signalling, among others. The results from individual genes and GSEA demonstrated that wt-VHSV impaired the activation at short stages of infection of pro-inflammatory, antiviral, proliferation, and apoptosis pathways, delaying innate humoral response and cellular crosstalk, whereas dNV-VHSV promoted the opposite effects. Therefore, these results might support future studies on using dNV-VHSV as a potential live vaccine.This research was partially funded by the Ministry of Science, Innovation and Universities of Spain grants AGL2014-51773-C3-3 and AGL2017-85494-C2-2-R, and the APC was funded by Ministry of Science, Innovation and Universities of Spain grant AGL2017-85494-C2-2-R.Peer reviewe

An ELISA for detection of trout antibodies to viral haemorrhagic septicemia virus using recombinant fragments of their viral G protein

An enzyme linked immunosorbent assay (ELISA) method to study serum antibodies to viral haemorrhagic septicemia virus (VHSV) was designed by using recombinant fragments of their G protein. By using this fragment-ELISA, we describe the binding of antibodies against recombinant G fragments of 45-445 amino acids present in VHSV-hyperimmunized trout sera. Fragments were designed by taking into account their tridimensional pH-dependent structure and functional domains. Sera were obtained from hyperimmunized trout following 4-5 intraperitoneal injections of VHSV antigens by using Freund's or saponin adjuvants. Sera from different hyperimmunized trout differed quantitatively rather than qualitatively in their recognition of solid-phase frg11 (56-110), frg12 (65-109), frg13 (97-167), frg14 (141-214), frg15 (65-250), frg16 (252-450) and G (G21-465) by Western blot and ELISA. However, titres were higher when using frg11, frg15 or frg16, rather than G21-465, suggesting higher accessibility to G epitopes. Further knowledge of the antigenicity of the G protein of rhabdoviruses by using fragments might be used to improve current vaccines. On the other hand, they might be used to dissect the trout antibody response to VHSV infections, to complement in vitro neutralizing assays, and/or to quantitate anti-VHSV antibodies in VHSV-infected/vaccinated trout, other fish and/or other body fluids such as mucus. © 2011 Elsevier B.V