Critical Role of FLRT1 Phosphorylation in the Interdependent Regulation of FLRT1 Function and FGF Receptor Signalling

Background Fibronectin leucine rich transmembrane (FLRT) proteins have dual properties as regulators of cell adhesion and potentiators of fibroblast growth factor (FGF) mediated signalling. The mechanism by which the latter is achieved is still unknown and is the subject of this investigation. Principal Findings Here we show that FLRT1 is a target for tyrosine phosphorylation mediated by FGFR1 and implicate a non-receptor Src family kinase (SFK). We identify the target tyrosine residues in the cytoplasmic domain of FLRT1 and show that these are not direct substrates for Src kinase suggesting that the SFK may exert effects via potentiation of FGFR1 kinase activity. We show that whilst FLRT1 expression results in a ligand-dependent elevation of MAP kinase activity, a mutant version of FLRT1, defective as an FGFR1 kinase substrate (Y3F-FLRT1), has the property of eliciting ligand-independent chronic activation of the MAP kinase pathway which is suppressed by pharmacological inhibition of either FGFR1 or Src kinase. Functional investigation of FGFR1 and FLRT1 signalling in SH-SY5Y neuroblastoma cells reveals that FLRT1 alone acts to induce a multi-polar phenotype whereas the combination of FLRT1 and FGFR activation, or expression of Y3F-FLRT1, acts to induce neurite outgrowth via MAPK activation. Similar results were obtained in a dendrite outgrowth assay in primary hippocampal neurons. We also show that FGFR1, FLRT1 and activated Src are co-localized and this complex is trafficked toward the soma of the cell. The presence of Y3F-FLRT1 rather than FLRT1 resulted in prolonged localization of this complex within the neuritic arbour. Conclusions This study shows that the phosphorylation state of FLRT1, which is itself FGFR1 dependent, may play a critical role in the potentiation of FGFR1 signalling and may also depend on a SFK-dependent phosphorylation mechanism acting via the FGFR. This is consistent with an ‘in vivo’ role for FLRT1 regulation of FGF signalling via SFKs. Furthermore, the phosphorylation-dependent futile cycle mechanism controlling FGFR1 signalling is concurrently crucial for regulation of FLRT1-mediated neurite outgrowth

Aeroelastic testing of LCA wing models - Model fabrication - Ground testing - Wind tunnel testing and Data analysis

Aeroelastic Testing Programme of Scaled Aeroelastic model of LCA half wing with rigid fuselage

Crystal structure and molecular imaging of the Nav channel β3 subunit indicates a trimeric assembly.

The vertebrate sodium (Nav) channel is composed of an ion-conducting α subunit and associated β subunits. Here, we report the crystal structure of the human β3 subunit immunoglobulin (Ig) domain, a functionally important component of Nav channels in neurons and cardiomyocytes. Surprisingly, we found that the β3 subunit Ig domain assembles as a trimer in the crystal asymmetric unit. Analytical ultracentrifugation confirmed the presence of Ig domain monomers, dimers, and trimers in free solution, and atomic force microscopy imaging also detected full-length β3 subunit monomers, dimers, and trimers. Mutation of a cysteine residue critical for maintaining the trimer interface destabilized both dimers and trimers. Using fluorescence photoactivated localization microscopy, we detected full-length β3 subunit trimers on the plasma membrane of transfected HEK293 cells. We further show that β3 subunits can bind to more than one site on the Nav 1.5 α subunit and induce the formation of α subunit oligomers, including trimers. Our results suggest a new and unexpected role for the β3 subunits in Nav channel cross-linking and provide new structural insights into some pathological Nav channel mutations

Basic science232. Certolizumab pegol prevents pro-inflammatory alterations in endothelial cell function

Background: Cardiovascular disease is a major comorbidity of rheumatoid arthritis (RA) and a leading cause of death. Chronic systemic inflammation involving tumour necrosis factor alpha (TNF) could contribute to endothelial activation and atherogenesis. A number of anti-TNF therapies are in current use for the treatment of RA, including certolizumab pegol (CZP), (Cimzia ®; UCB, Belgium). Anti-TNF therapy has been associated with reduced clinical cardiovascular disease risk and ameliorated vascular function in RA patients. However, the specific effects of TNF inhibitors on endothelial cell function are largely unknown. Our aim was to investigate the mechanisms underpinning CZP effects on TNF-activated human endothelial cells. Methods: Human aortic endothelial cells (HAoECs) were cultured in vitro and exposed to a) TNF alone, b) TNF plus CZP, or c) neither agent. Microarray analysis was used to examine the transcriptional profile of cells treated for 6 hrs and quantitative polymerase chain reaction (qPCR) analysed gene expression at 1, 3, 6 and 24 hrs. NF-κB localization and IκB degradation were investigated using immunocytochemistry, high content analysis and western blotting. Flow cytometry was conducted to detect microparticle release from HAoECs. Results: Transcriptional profiling revealed that while TNF alone had strong effects on endothelial gene expression, TNF and CZP in combination produced a global gene expression pattern similar to untreated control. The two most highly up-regulated genes in response to TNF treatment were adhesion molecules E-selectin and VCAM-1 (q 0.2 compared to control; p > 0.05 compared to TNF alone). The NF-κB pathway was confirmed as a downstream target of TNF-induced HAoEC activation, via nuclear translocation of NF-κB and degradation of IκB, effects which were abolished by treatment with CZP. In addition, flow cytometry detected an increased production of endothelial microparticles in TNF-activated HAoECs, which was prevented by treatment with CZP. Conclusions: We have found at a cellular level that a clinically available TNF inhibitor, CZP reduces the expression of adhesion molecule expression, and prevents TNF-induced activation of the NF-κB pathway. Furthermore, CZP prevents the production of microparticles by activated endothelial cells. This could be central to the prevention of inflammatory environments underlying these conditions and measurement of microparticles has potential as a novel prognostic marker for future cardiovascular events in this patient group. Disclosure statement: Y.A. received a research grant from UCB. I.B. received a research grant from UCB. S.H. received a research grant from UCB. All other authors have declared no conflicts of interes

The Geomagnetic cavity and the Van Allen radiation belts-II

Using the spherical harmonic expansion given by Mead for the magnetic field created by the currents on the surface of the geomagnetic cavity, the nature of the trapping regions has been discussed. It is shown that to a first order of approximation in the expansion coefficients, the allowed regions are still determined by a cubic equation. Numerical determinations have been made for the structure of the allowed regions for particles of different energies both on the sunlit side as well as the dark side of the Earth.13

Regulation of AtKUP2 Expression by bHLH and WRKY Transcription Factors Helps to Confer Increased Salt Tolerance to Arabidopsis thaliana Plants

10.3389/fpls.2020.01311Frontiers in Plant Science11131

Design and implementation of a novel architecture for physical human-UAV interaction

Interaction between humans and unmanned aerial vehicles is a promising field for future applications. However, current interfacing paradigms either imply the presence of intermediary hardware as monitors, joysticks and haptic devices, or are limited to visual/auditory channels with hand gestures, voice recognition, or interpretation of face poses and body postures. Another paradigm, physical human–robot interaction, which is based on mutual exchange of forces, is popular when dealing with robotic arms and humanoids, while unmanned aerial vehicles are usually considered too dangerous and lack proper interaction surfaces to exchange forces. In this paper, we address the problem of physical human–unmanned aerial vehicle interaction and we propose a straightforward approach to allow a human to intuitively command an unmanned aerial vehicle through exchanges of forces. Using a residual based estimator, we estimate the external forces and torques acting on the unmanned aerial vehicle. Through the employment of a sensor ring, we are able to separate the human interaction forces from additional disturbances as wind and parameter uncertainties. This knowledge is used inside a control framework where the human is allowed to change the desired trajectory by simply applying forces on the unmanned aerial vehicle. The system is validated with multiple hardware-in-the-loop simulations and experiments in which we try different interaction modalities

A Control Architecture for Physical Human-UAV Interaction with a Fully Actuated Hexarotor

Physical human-UAV interaction (PHUI) with Unmanned Aerial Vehicles (UAV) has many possible applications, and a few recent works have shown that it is possible to perform PHUI with a standard quadrotor. However, the intrinsic underactuation of quadrotors may hinder the interaction task and also cause safety issues. In this paper, we present an admittance control-based scheme to perform PHUI with a fully actuated UAV. The system also benefits from the robustness provided by our fully-actuated-UAV implementation of the adaptive super twisting controller (ASTC). While validating our system in simulation, we also show the superior performance of the fully actuated UAV with respect to a standard quadcopter