Differential contribution of PB1-F2 to the virulence of highly pathogenic H5N1 influenza A virus in mammalian and avian species

Highly pathogenic avian influenza A viruses (HPAIV) of the H5N1 subtype occasionally transmit from birds to humans and can cause severe systemic infections in both hosts. PB1-F2 is an alternative translation product of the viral PB1 segment that was initially characterized as a pro-apoptotic mitochondrial viral pathogenicity factor. A full-length PB1-F2 has been present in all human influenza pandemic virus isolates of the 20(th) century, but appears to be lost evolutionarily over time as the new virus establishes itself and circulates in the human host. In contrast, the open reading frame (ORF) for PB1-F2 is exceptionally well-conserved in avian influenza virus isolates. Here we perform a comparative study to show for the first time that PB1-F2 is a pathogenicity determinant for HPAIV (A/Viet Nam/1203/2004, VN1203 (H5N1)) in both mammals and birds. In a mammalian host, the rare N66S polymorphism in PB1-F2 that was previously described to be associated with high lethality of the 1918 influenza A virus showed increased replication and virulence of a recombinant VN1203 H5N1 virus, while deletion of the entire PB1-F2 ORF had negligible effects. Interestingly, the N66S substituted virus efficiently invades the CNS and replicates in the brain of Mx+/+ mice. In ducks deletion of PB1-F2 clearly resulted in delayed onset of clinical symptoms and systemic spreading of virus, while variations at position 66 played only a minor role in pathogenesis. These data implicate PB1-F2 as an important pathogenicity factor in ducks independent of sequence variations at position 66. Our data could explain why PB1-F2 is conserved in avian influenza virus isolates and only impacts pathogenicity in mammals when containing certain amino acid motifs such as the rare N66S polymorphism

A Gravitational Wave Background from Reheating after Hybrid Inflation

The reheating of the universe after hybrid inflation proceeds through the nucleation and subsequent collision of large concentrations of energy density in the form of bubble-like structures moving at relativistic speeds. This generates a significant fraction of energy in the form of a stochastic background of gravitational waves, whose time evolution is determined by the successive stages of reheating: First, tachyonic preheating makes the amplitude of gravity waves grow exponentially fast. Second, bubble collisions add a new burst of gravitational radiation. Third, turbulent motions finally sets the end of gravitational waves production. From then on, these waves propagate unimpeded to us. We find that the fraction of energy density today in these primordial gravitational waves could be significant for GUT-scale models of inflation, although well beyond the frequency range sensitivity of gravitational wave observatories like LIGO, LISA or BBO. However, low-scale models could still produce a detectable signal at frequencies accessible to BBO or DECIGO. For comparison, we have also computed the analogous gravitational wave background from some chaotic inflation models and obtained results similar to those found by other groups. The discovery of such a background would open a new observational window into the very early universe, where the details of the process of reheating, i.e. the Big Bang, could be explored. Moreover, it could also serve in the future as a new experimental tool for testing the Inflationary Paradigm.Comment: 22 pages, 18 figures, uses revtex

Highly photostable solid-state dye lasers based on silicon-modified organic matrices

11 pages, 13 figures, 4 tables.-- PACS: 42.55.Rz; 42.60.FcWe report on the synthesis, characterization, and physical properties of modified polymeric matrices incorporating silicon atoms in their structure and doped with laser dyes. When the silicon-modified organic matrices incorporated pyrromethene 567 (PM567) and pyrromethene 597 (PM597) dyes as actual solid solutions, highly photostable laser operation with reasonable, nonoptimized efficiencies was obtained under transversal pumping at 532 nm. At a pump repetition rate of 10 Hz, the intensity of the laser emission remained at the level or above the initial lasing intensity after 100 000 pump pulses in the same position of the sample, corresponding to an estimated accumulated pump energy absorbed by the system of 518 and 1295 GJ/mol for PM567 and PM597, respectively. When the pump repetition rate was increased to 30 Hz, the laser emission of dye PM567 decreased steadily and the output energy fell to one-half its initial value after an accumulated pump energy of 989 GJ/mol. Dye PM597 demonstrated a remarkable photostability, and under 30 Hz pumping the laser emission from some samples remained stable after 700 000 pump pulses in the same position of the sample, corresponding to an accumulated pump energy of 17 300 GJ/mol. Narrow linewidth operation with tuning ranges of up to 31 nm was obtained with both pyrromethene dyes when some of the samples were incorporated into a grazing-incidence grating oscillator.This work was supported by Project Nos. 7N/0100/02 of the Comunidad Autónoma de Madrid ( CAM) and MAT2004- 04643-C03-01 of the Spanish CICYT. One of the authors (O.G.) thanks the MEC for awarding her a Ramón y Cajal scientific contract. Another author (D.A.) thanks CAM for a predoctoral scholarship.Peer reviewe

GSE4, a small dyskerin- and GSE24.2-related peptide, induces telomerase activity, cell proliferation and reduces DNA damage, oxidative stress and cell senescence in dyskerin mutant cells

Dyskeratosis congenita is an inherited disease caused by mutations in genes coding for telomeric components. It was previously reported that expression of a dyskerin-derived peptide, GSE24.2, increases telomerase activity, regulates gene expression and decreases DNA damage and oxidative stress in dyskeratosis congenita patient cells. The biological activity of short peptides derived from GSE24.2 was tested and one of them, GSE4, that probed to be active, was further characterized in this article. Expression of this eleven amino acids long peptide increased telomerase activity and reduced DNA damage, oxidative stress and cell senescence in dyskerin-mutated cells. GSE4 expression also activated c-myc and TERT promoters and increase of c-myc, TERT and TERC expression. The level of biological activity of GSE4 was similar to that obtained by GSE24.2 expression. Incorporation of a dyskerin nuclear localization signal to GSE24.2 did not change its activity on promoter regulation and DNA damage protection. However, incorporation of a signal that increases the rate of nucleolar localization impaired GSE24.2 activity. Incorporation of the dyskerin nuclear localization signal to GSE4 did not alter its biological activity. Mutation of the Aspartic Acid residue that is conserved in the pseudouridine synthase domain present in GSE4 did not impair its activity, except for the repression of c-myc promoter activity and the decrease of c-myc, TERT and TERC gene expression in dyskerin-mutated cells. These results indicated that GSE4 could be of great therapeutic interest for treatment of dyskeratosis congenita patients.This work was supported by grants PI1401495 (supported by FEDER funds) and ER15PR07ACC114/757 (Fondo de Investigaciones Sanitarias, Instituto de Salud Carlos III. Spain), 201320E075 (Consejo Superior de Investigaciones Científicas) and IPT-2012-0674- 090000 (Ministerio de Economía y Competitividad. Spain). CM-G is supported by the CIBER de Enfermedades Raras.Peer Reviewe

Host-Specific NS5 Ubiquitination Determines Yellow Fever Virus Tropism

The recent yellow fever virus (YFV) epidemic in Brazil in 2017 and Zika virus (ZIKV) epidemic in 2015 serve to remind us of the importance of flaviviruses as emerging human pathogens. With the current global flavivirus threat, there is an urgent need for antivirals and vaccines to curb the spread of these viruses. However, the lack of suitable animal models limits the research questions that can be answered. A common trait of all flaviviruses studied thus far is their ability to antagonize interferon (IFN) signaling so as to enhance viral replication and dissemination. Previously, we reported that YFV NS5 requires the presence of type I IFN (IFN-α/β) for its engagement with human signal transducer and activator of transcription 2 (hSTAT2). In this manuscript, we report that like the NS5 proteins of ZIKV and dengue virus (DENV), YFV NS5 protein is able to bind hSTAT2 but not murine STAT2 (mSTAT2). Contrary to what has been demonstrated with ZIKV NS5 and DENV NS5, replacing mSTAT2 with hSTAT2 cannot rescue the YFV NS5-STAT2 interaction, as YFV NS5 is also unable to interact with hSTAT2 in murine cells. We show that the IFN-α/β-dependent ubiquitination of YFV NS5 that is required for STAT2 binding in human cells is absent in murine cells. In addition, we demonstrate that mSTAT2 restricts YFV replication in vivo These data serve as further impetus for the development of an immunocompetent mouse model that can serve as a disease model for multiple flaviviruses.IMPORTANCE Flaviviruses such as yellow fever virus (YFV), Zika virus (ZIKV), and dengue virus (DENV) are important human pathogens. A common flavivirus trait is the antagonism of interferon (IFN) signaling to enhance viral replication and spread. We report that like ZIKV NS5 and DENV NS5, YFV NS5 binds human STAT2 (hSTAT2) but not mouse STAT2 (mSTAT2), a type I IFN (IFN-α/β) pathway component. Additionally, we show that contrary to what has been demonstrated with ZIKV NS5 and DENV NS5, YFV NS5 is unable to interact with hSTAT2 in murine cells. We demonstrate that mSTAT2 restricts YFV replication in mice and that this correlates with a lack of IFN-α/β-induced YFV NS5 ubiquitination in murine cells. The lack of suitable animal models limits flavivirus pathogenesis, vaccine, and drug research. These data serve as further impetus for the development of an immunocompetent mouse model that can serve as a disease model for multiple flaviviruses

A universal influenza virus vaccine candidate confers protection against pandemic H1N1 infection in ferrets

Influenza viruses can cause severe disease and mortality in humans. Due to constant change in their immuno-dominant antigenic sites they can evade adaptive immune responses. Current seasonal influenza virus vaccines therefore require annual re-formulation and re-administration to confer protection from circulating viruses. Additionally, these vaccines cannot protect against novel pandemic influenza virus strains. Novel vaccination approaches attempt to refocus antibody responses towards more conserved domains like the hemagglutinin stalk. Antibodies against the stalk domain are broadly-reactive and can neutralize multiple influenza virus subtypes. However, the stalk domain is immuno-subdominant and not preferentially targeted by the immune system. In this study, we tested if a vaccination strategy based on influenza viruses expressing chimeric hemagglutinins (cH) that contain exotic, divergent head domains, but a conserved H1 stalk domain could induce cross-protective antibody responses in ferrets. We compared a heterologous live-attenuated virus (cH8/1N1) prime followed by an inactivated split virus (cH5/1N1) boost combination approach to two doses of split-virus vaccines (cH8/1N1/cH5/1N1) and the impact of adjuvant on the immune response. Additionally, a ‘standard of care’ control group received 2 rounds of a human trivalent influenza virus vaccine. We found that all universal influenza virus vaccination approaches were successful at inducing stalk-reactive antibody responses in serum. Virus replication was limited to the nasopharynx in the live attenuated/split vaccine groups and nasal wash titers were lower than in the \u27standard of care\u27 control group. No virus replication was detected in the lungs of attenuated/split vaccinated ferrets, while the \u27standard of care\u27 group had similarly high titers as an unvaccinated control group. Our findings demonstrate that - using a chimeric hemagglutinin based heterologous attenuated/split combination strategy - our candidate universal influenza virus vaccine can successfully protect ferrets from pandemic H1N1 infection. The data support further development of this vaccination approach and advancement into clinical trials

Impact of COVID-19 lockdown in telecommunications engineering competency-based alumni ranking

[EN] Higher education, as other social and economic sectors in Spain, was disrupted on 15th March 2020 following health emergency Laws enacted related to COVID-19. Non-presential lecturing was stablished country-wide until the end of the academic year 2019-20, as society was set in lockdown. In this context, it is necessary to evaluate the impact of the mandatory changes implemented in the education paradigm in order to assess the degree of acquisition of general and specific competencies altogether the acquisition of transversal competencies, as an important factor for the alumni career. This paper reports a comprehensive study on the competencies degree of acquisition considering the lockdown scenario in Spain. The results from the last four academic years have been comparatively evaluated in the core subject `Teoría de la Comunicación¿, lectured in the fourth semester of the Telecommunications Engineering Integrated Program (Bachelor and Master) in the Universitat Politècnica de València, Spain, comprising data from 745 alumni. The results indicate that the degree of acquisition of technical competencies in this scenario has been adequate, being marginally better compared to previous academic year, probably due to new lecturing materials prepared. Nevertheless, the acquisition of the transversal competency `analysis and problem solving¿ exhibits degraded results, indicating inhomogeneous acquisition probably due to limitations in the group-based problem-solving practice. The results suggest that specific materials and remote lecturing strategies should be developed and implemented to guarantee adequate acquisition levels.The support by the 2020 Science Parks program from the Consellería de Innovación, Universidades, Ciencia y Sociedad Digital, Generalitat Valenciana, Spain, is acknowledged.Llorente, R.; Rodríguez-Hernández, MA.; Hernandez Franco, CA.; Sastre, J.; Carrión García, A.; Madrigal-Madrigal, J. (2020). Impact of COVID-19 lockdown in telecommunications engineering competency-based alumni ranking. IATED Academy. 9599-9607. https://doi.org/10.21125/iceri.2020.2139S9599960

The pause on avian H5N1 influenza virus transmission research should be ended

A voluntary 60-day pause on avian H5N1 influenza virus transmission research was announced in January 2012 by the international community of influenza scientists engaged in this work to provide time to explain the benefits of such work and the risk mitigation measures in place. Subsequently, the pause was extended to allow for time for review of the biosafety and bios-ecurity conditions. After almost 8 months, these conditions have been met in some countries and are close to being met in others. Because H5N1 virus transmission studies are essential for pandemic preparedness, researchers who have approval from their governments and institutions to conduct this research safely under appropriate biosecurity conditions should resume this important work

Selective Pressure by Rifampicin Modulates Mutation Rates and Evolutionary Trajectories of Mycobacterial Genomes

Resistance to the frontline antibiotic rifampicin constitutes a challenge to the treatment and control of tuberculosis. Here, we analyzed the mutational landscape of Mycobacterium smegmatis during long-term evolution with increasing concentrations of rifampicin, using a mutation accumulation assay combined with whole-genome sequencing. Antibiotic treatment enhanced the acquisition of mutations, doubling the genome-wide mutation rate of the wild-type cells. While antibiotic exposure led to extinction of almost all wild-type lines, the hypermutable phenotype of the ΔnucS mutant strain (noncanonical mismatch repair deficient) provided an efficient response to the antibiotic, leading to high rates of survival. This adaptative advantage resulted in the emergence of higher levels of rifampicin resistance, an accelerated acquisition of drug resistance mutations in rpoB (β RNA polymerase), and a wider diversity of evolutionary pathways that led to drug resistance. Finally, this approach revealed a subset of adaptive genes under positive selection with rifampicin that could be associated with the development of antibiotic resistance. IMPORTANCE Rifampicin is the most important first-line antibiotic against mycobacterial infections, including tuberculosis, one of the top causes of death worldwide. Acquisition of rifampicin resistance constitutes a major global public health problem that makes the control of the disease challenging. Here, we performed an experimental evolution assay under antibiotic selection to analyze the response and adaptation of mycobacteria, leading to the acquisition of rifampicin resistance. This approach explored the total number of mutations that arose in the mycobacterial genomes under long-term rifampicin exposure, using whole-genome sequencing. Our results revealed the effect of rifampicin at a genomic level, identifying different mechanisms and multiple pathways leading to rifampicin resistance in mycobacteria. Moreover, this study detected that an increase in the rate of mutations led to enhanced levels of drug resistance and survival. In summary, all of these results could be useful to understand and prevent the emergence of drug-resistant isolates in mycobacterial infections.This research was funded by MCIN/AEI/10.13039/501100011033, grant PID2020-112865RB-I00, and Instituto de Salud Carlos III, grant FIS PI17/00159 (ISCIII/FEDER, UE). E.C.-S. is the recipient of a PFIS predoctoral research fellowship (FI18/00036) cofinanced by the Instituto de Salud Carlos III and the European Social Fund. A.C.-G. acknowledges financial support from the Spanish State Research Agency, AEI/10.13039/501100011033, through the “Severo Ochoa” Program for Centers of Excellence in R&D (SEV-2013-0347, SEV-2017-0712). Editorial assistance was provided by Stuart L. Rulten. Statistical consultancy was provided by Applied Statistical Department-SGAI-CSIC.S