0341: AMPK exerts an insulin-sensitizing effect on cardiac glucose uptake by multiple molecular mechanisms including cytoskeleton reorganization

BackgroundInsulin-resistant cardiomyocytes are characterized by a decreased ability of insulin to stimulate glucose uptake. We have previously shown that the activation of AMPK by metformin or phenformin restores insulin-sensitivity in insulin-resistant cardiomyocytes. The aim of our present work is to understand by which molecular mechanisms AMPK exerts its insulin sensitizing effect. In this study we focused on the mTOR/p70S6K pathway and on cytoskeleton reorganization. mTOR/p70S6K, which is known to be inhibited by AMPK, is able to reduce insulin signaling via a negative feedback loop involving serine phosphorylation of IRS-1. On the other hand, cytoskeleton reorganization, which is a known target of AMPK, is responsible for the translocation of the glucose transporter GLUT4 to the plasma membrane.MethodsAdult rat cardiomyocytes were primary cultured and treated with different agents including insulin, AMPK activator (phenformin), mTOR inhibitor rapamycin and/or actin cytoskeleton inhibitor latrunculin B. Glucose uptake was assessed by detritiation of 2-3H-glucose.ResultsFirst, we tested if rapamycin was able to mimic AMPK activators. Similarly to phenformin, rapamycin increased the insulin-dependent phosphorylation of Akt involved in the regulation of glucose uptake. Despite the ability of rapamycin to induce this Akt over-phosphorylation, rapamycin was not able to restore the insulin-dependent stimulation of glucose uptake like phenformin did. On the other hand, latrunculin B abolished the insulin-sensitizing action of phenformin on glucose uptake, in insulin-sensitive as well as in insulinresistant cells.Conclusionsactin cytoskeleton reorganization but not mTOR/p70S6K, is involved in the insulin-sensitizing effect of AMPK on cardiac glucose uptake. The role played by Small G proteins, known to be involved in the regulation of actin cytoskeleton is under investigation

Non-Planar Pad-Printed Think-Film Focused High-Frequency Ultrasonic Transducers for Imaging and Therapeutic Applications

Pad-printed thick-film transducers have been shown to be an interesting alternative to lapped bulk piezoceramics, because the film is deposited with the required thickness, size, and geometry, thus avoiding any subsequent machining to achieve geometrical focusing. Their electromechanical properties are close to those of bulk ceramics with similar composition despite having a higher porosity. In this paper, padprinted high-frequency transducers based on a low-loss piezoceramic composition are designed and fabricated. High-porosity ceramic cylinders with a spherical top surface are used as the backing substrate. The transducers are characterized in view of imaging applications and their imaging capabilities are evaluated with phantoms containing spherical inclusions and in different biological tissues. In addition, the transducers are evaluated for their capability to produce high-acoustic intensities at frequencies around 20 MHz. High-intensity measurements, obtained with a calibrated hydrophone, show that transducer performance is promising for applications that would require the same device to be used for imaging and for therapy. Nevertheless, the transducer design can be improved, and simulation studies are performed to find a better compromise between low-power and high-power performance. The size, geometry, and constitutive materials of optimized configurations are proposed and their feasibility is discussed

The Io, Europa and Ganymede auroral footprints at Jupiter in the ultraviolet: positions and equatorial lead angles

Jupiter's satellite auroral footprints are a consequence of the interaction between the Jovian magnetic field with co-rotating iogenic plasma and the Galilean moons. The disturbances created near the moons propagate as Alfv\'en waves along the magnetic field lines. The position of the moons is therefore "Alfv\'enically" connected to their respective auroral footprint. The angular separation from the instantaneous magnetic footprint can be estimated by the so-called lead angle. That lead angle varies periodically as a function of orbital longitude, since the time for the Alfv\'en waves to reach the Jovian ionosphere varies accordingly. Using spectral images of the Main Alfv\'en Wing auroral spots collected by Juno-UVS during the first forty-three orbits, this work provides the first empirical model of the Io, Europa and Ganymede equatorial lead angles for the northern and southern hemispheres. Alfv\'en travel times between the three innermost Galilean moons to Jupiter's northern and southern hemispheres are estimated from the lead angle measurements. We also demonstrate the accuracy of the mapping from the Juno magnetic field reference model (JRM33) at the completion of the prime mission for M-shells extending to at least 15RJ . Finally, we shows how the added knowledge of the lead angle can improve the interpretation of the moon-induced decametric emissions.Comment: 20 pages, 8 figures, Accepted for publication in Journal of Geophysical Research: Space Physics on 20 April 202

Cytosolic PLA2 is required for CTL-mediated immunopathology of celiac disease via NKG2D and IL-15

IL-15 and NKG2D promote autoimmunity and celiac disease by arming cytotoxic T lymphocytes (CTLs) to cause tissue destruction. However, the downstream signaling events underlying these functional properties remain unclear. Here, we identify cytosolic phospholipase A2 (cPLA2) as a central molecule in NKG2D-mediated cytolysis in CTLs. Furthermore, we report that NKG2D induces, upon recognition of MIC+ target cells, the release of arachidonic acid (AA) by CTLs to promote tissue inflammation in association with target killing. Interestingly, IL-15, which licenses NKG2D-mediated lymphokine killer activity in CTLs, cooperates with NKG2D to induce cPLA2 activation and AA release. Finally, cPLA2 activation in intraepithelial CTLs of celiac patients provides an in vivo pathophysiological dimension to cPLA2 activation in CTLs. These results reveal an unrecognized link between NKG2D and tissue inflammation, which may underlie the emerging role of NKG2D in various immunopathological conditions and define new therapeutic targets

IgG and Fcγ Receptors in Intestinal Immunity and Inflammation.

Fcγ receptors (FcγR) are cell surface glycoproteins that mediate cellular effector functions of immunoglobulin G (IgG) antibodies. Genetic variation in FcγR genes can influence susceptibility to a variety of antibody-mediated autoimmune and inflammatory disorders, including systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). More recently, however, genetic studies have implicated altered FcγR signaling in the pathogenesis of inflammatory bowel disease (IBD), a condition classically associated with dysregulated innate and T cell immunity. Specifically, a variant of the activating receptor, FcγRIIA, with low affinity for IgG, confers protection against the development of ulcerative colitis, a subset of IBD, leading to a re-evaluation of the role of IgG and FcγRs in gastrointestinal tract immunity, an organ system traditionally associated with IgA. In this review, we summarize our current understanding of IgG and FcγR function at this unique host-environment interface, from the pathogenesis of colitis and defense against enteropathogens, its contribution to maternal-fetal cross-talk and susceptibility to cancer. Finally, we discuss the therapeutic implications of this information, both in terms of how FcγR signaling pathways may be targeted for the treatment of IBD and how FcγR engagement may influence the efficacy of therapeutic monoclonal antibodies in IBD