FDA-approved Abl/EGFR/PDGFR kinase inhibitors show potent efficacy against pandemic and seasonal influenza A virus infections of human lung explants.

Influenza viruses (IVs) cause substantial global morbidity and mortality. Given the limited range of licensed antiviral drugs and their reduced efficacy due to resistance mutations, repurposing FDA-approved kinase inhibitors as fast-tracked host-targeted antivirals is an attractive strategy. We identified six FDA-approved non-receptor tyrosine kinase-inhibitors (NRTKIs) as potent inhibitors of viral replication of pandemic and seasonal IVs in vitro. We validated their efficacy in a biologically and clinically relevant ex vivo model of human precision-cut lung slices. We identified steps of the virus infection cycle affected by these inhibitors and assessed their effect(s) on host responses. Their overlapping targets suggest crosstalk between Abl, EGFR, and PDGFR pathways during IAV infection. Our data and established safety profiles of these NRTKIs provide compelling evidence for further clinical investigations and repurposing as host-targeted influenza antivirals. Moreover, these NRTKIs have broad-spectrum antiviral potential given that their kinase/pathway targets are critical for the replication of many viruses.

FDA-Approved Inhibitors of RTK/Raf Signaling Potently Impair Multiple Steps of In Vitro and Ex Vivo Influenza A Virus Infections.

Influenza virus (IV) infections pose a burden on global public health with significant morbidity and mortality. The limited range of currently licensed IV antiviral drugs is susceptible to the rapid rise of resistant viruses. In contrast, FDA-approved kinase inhibitors can be repurposed as fast-tracked host-targeted antivirals with a higher barrier of resistance. Extending our recent studies, we screened 21 FDA-approved small-molecule kinase inhibitors (SMKIs) and identified seven candidates as potent inhibitors of pandemic and seasonal IV infections. These SMKIs were further validated in a biologically and clinically relevant ex vivo model of human precision-cut lung slices. We identified steps of the virus infection cycle affected by these inhibitors (entry, replication, egress) and found that most SMKIs affected both entry and egress. Based on defined and overlapping targets of these inhibitors, the candidate SMKIs target receptor tyrosine kinase (RTK)-mediated activation of Raf/MEK/ERK pathways to limit influenza A virus infection. Our data and the established safety profiles of these SMKIs support further clinical investigations and repurposing of these SMKIs as host-targeted influenza therapeutics.

Recovery during and after a simulated multi-day tennis tournament: Combining active recovery, stretching, cold-water immersion, and massage interventions.

The aim of this study was to investigate the effects of a mixed-method recovery intervention (MMR) consisting of active recovery, stretching, cold-water immersion, and massage on physical, technical, physiological, and perceptual recovery during and after a five-day simulated tennis tournament. Nine competitive male tennis players (age, 24.6±4.2 years) with national ranking positions (German Tennis Federation) and Universal Tennis Ratings between approximately 11-13 participated in two singles tennis tournaments, which were separated by a three-month washout period. During the tournaments, participants played five two-and-a-half-hour competitive singles tennis match on five consecutive days. For the assignment to one of two groups, athletes were matched into homogeneous pairs according to their ranking. Then, within each pair, the players were randomly assigned to one of two groups. The first group performed MMR during the first tournament, whereas the other group used passive recovery (PAS). During the second tournament, recovery conditions were interchanged. Measures of physical and technical performance as well as physiological and perceptual responses (heart rate, blood lactate concentration, perceived exertion) were recorded during match-play sessions. Furthermore, muscle soreness, perceived recovery state, blood markers, countermovement jump height (CMJ), and repeated sprint ability (RSA) were determined before, during, and after the five-day tournament periods. Results showed significant changes over time (P < 0.05) in muscle soreness, perceived recovery state, creatine kinase, c-reactive protein, insulin-like growth factor 1, and countermovement jump height. However, no significant differences or recovery strategy x time interactions were noted either for tennis-specific performance (e.g. number of total points won) or any other of the measured parameters between MMR and PAS (P > 0.05). In conclusion, the repeated use of MMR during and after a five-day tennis tournament did not affect match performance, match load, or recovery from repeated days of tennis match play.

The floral regulator LEAFY evolves by substitutions in the DNA binding domain.

The plant-specific transcription factor LEAFY controls general aspects of the life cycle in a basal plant, the moss Physcomitrella patens. In contrast, LEAFY has more specialized functions in angiosperms, where it specifically induces floral fate during the reproductive phase. This raises the question of a concomitant change in the biochemical function of LEAFY during the evolution of land plants. We report that the DNA binding domain of LEAFY, although largely conserved, has diverged in activity. On the contrary, other, more rapidly evolving portions of the protein have few effects on LEAFY activity.

Regulatory elements of the floral homeotic gene AGAMOUS identified by phylogenetic footprinting and shadowing.

In Arabidopsis thaliana, cis-regulatory sequences of the floral homeotic gene AGAMOUS (AG) are located in the second intron. This 3-kb intron contains binding sites for two direct activators of AG, LEAFY (LFY) and WUSCHEL (WUS), along with other putative regulatory elements. We have used phylogenetic footprinting and the related technique of phylogenetic shadowing to identify putative cis-regulatory elements in this intron. Among 29 Brassicaceae species, several other motifs, but not the LFY and WUS binding sites identified previously, are largely invariant. Using reporter gene analyses, we tested six of these motifs and found that they are all functionally important for the activity of AG regulatory sequences in A. thaliana. Although there is little obvious sequence similarity outside the Brassicaceae, the intron from cucumber AG has at least partial activity in A. thaliana. Our studies underscore the value of the comparative approach as a tool that complements gene-by-gene promoter dissection but also demonstrate that sequence-based studies alone are insufficient for a complete identification of cis-regulatory sites.