A CACNA1C variant associated with cardiac arrhythmias provides mechanistic insights in the calmodulation of L-type Ca2+ channels

We recently reported the identification of a de novo single nucleotide variant in exon 9 of CACNA1C associated with prolonged repolarization interval. Recombinant expression of the glycine to arginine variant at position 419 produced a gain in the function of the L-type CaV1.2 channel with increased peak current density and activation gating but without signif- icant decrease in the inactivation kinetics. We herein reveal that these properties are replicated by overexpressing calmodulin (CaM) with CaV1.2 WT and are reversed by expo- sure to the CaM antagonist W-13. Phosphomimetic (T79D or S81D), but not phosphoresistant (T79A or S81A), CaM surro- gates reproduced the impact of CaM WT on the function of CaV1.2 WT. The increased channel activity of CaV1.2 WT following overexpression of CaM was found to arise in part from enhanced cell surface expression. In contrast, the prop- erties of the variant remained unaffected by any of these treatments. CaV1.2 substituted with the α-helix breaking pro- line residue were more reluctant to open than CaV1.2 WT but were upregulated by phosphomimetic CaM surrogates. Our results indicate that (1) CaM and its phosphomimetic analogs promote a gain in the function of CaV1.2 and (2) the structural properties of the first intracellular linker of CaV1.2 contribute to its CaM-induced modulation. We conclude that the CAC- NA1C clinical variant mimics the increased activity associated with the upregulation of CaV1.2 by Ca2+–CaM, thus main- taining a majority of channels in a constitutively active mode that could ultimately promote ventricular arrhythmias

Identification of glycosylation sites essential for surface expression of the Caᵥα2δ1 subunit and modulation of the Cardiac Caᵥ1.2 channel activity

Alteration in the L-type current density is one aspect of the electrical remodeling observed in patients suffering from cardiac arrhythmias. Changes in channel function could result from variations in the protein biogenesis, stability, post-translational modification, and/or trafficking in any of the regulatory subunits forming cardiac L-type Ca2+ channel complexes. CaVα2δ1 is potentially the most heavily N-glycosylated subunit in the cardiac L-type CaV1.2 channel complex. Here, we show that enzymatic removal of N-glycans produced a 50-kDa shift in the mobility of cardiac and recombinant CaVα2δ1 proteins. This change was also observed upon simultaneous mutation of the 16 Asn sites. Nonetheless, the mutation of only 6/16 sites was sufficient to significantly 1) reduce the steady-state cell surface fluorescence of CaVα2δ1 as characterized by two-color flow cytometry assays and confocal imaging; 2) decrease protein stability estimated from cycloheximide chase assays; and 3) prevent the CaVα2δ1-mediated increase in the peak current density and voltage-dependent gating of CaV1.2. Reversing the N348Q and N812Q mutations in the non-operational sextuplet Asn mutant protein partially restored CaVα2δ1 function. Single mutation N663Q and double mutations N348Q/N468Q, N348Q/N812Q, and N468Q/N812Q decreased protein stability/synthesis and nearly abolished steady-state cell surface density of CaVα2δ1 as well as the CaVα2δ1-induced up-regulation of L-type currents. These results demonstrate that Asn-663 and to a lesser extent Asn-348, Asn-468, and Asn-812 contribute to protein stability/synthesis of CaVα2δ1, and furthermore that N-glycosylation of CaVα2δ1 is essential to produce functional L-type Ca2+ channels

PODIUM:A Pulsar Navigation Unit for Science Missions

PODIUM is a compact spacecraft navigation unit, currently being designed to provide interplanetary missions with autonomous position and velocity estimations. The unit will make use of Pulsar X-ray observations to measure the distance and distance rate from the host spacecraft to the Solar System Barycenter. Such measurements will then be used by the onboard orbit determination function to estimate the complete orbital elements of the spacecraft. The design aims at 6 kg of mass and 20 W of power, in a volume of 150 mm by 240 mm by 600 mm. PODIUM is designed to minimize the impact on the mission operational and accommodation constraints. The architecture is based on a grazing incidence X-ray telescope with focal distance limited to 50 cm. The effective area shall be in the range 25 to 50 cm2 for photon energies in the range 0.2-10 keV, requiring nesting of several mirrors in the Wolter-1 geometry. Grazing incidence angles will be very small, below 2 deg. The current target FOV is 0.25 deg. The pulsars photon arrivals are detected with a single pixel Silicon Drift Detector (SDD) sensor with timing accuracy below 1usec. The unit has no gimbaling to meet the applicable power, size and mass requirements. Instead, the host spacecraft shall slew and point to allow pulsar observation. The avionics architecture is based on a radiation hardened LEON4 processor, to allow a synchronous propagation task and measurement generation and orbit determination step in an asynchronous task. PODIUM will enable higher autonomy and lower cost for interplanetary missions. L2 space observatories and planetary flybys are the current reference use cases. Onboard autonomous state estimation can reduce the ground support effort required for navigation and orbit correction/maintenance computation, and reduce the turnaround time, thus enabling more accurate maneuvers, reducing the orbit maintenance mass budget

Constraining the Nature of the 18 min Periodic Radio Transient GLEAM-X J162759.5-523504.3 via Multiwavelength Observations and Magneto-thermal Simulations

We observed the periodic radio transient GLEAM-X J162759.5-523504.3 (GLEAM-X J1627) using the Chandra X-ray Observatory for about 30 ks on 2022 January 22–23, simultaneously with radio observations from the Murchison Widefield Array, MeerKAT, and the Australia Telescope Compact Array. Its radio emission and 18 min periodicity led the source to be tentatively interpreted as an extreme magnetar or a peculiar highly magnetic white dwarf. The source was not detected in the 0.3–8 keV energy range with a 3σ upper limit on the count rate of 3 × 10−4 counts s−1. No radio emission was detected during our X-ray observations either. Furthermore, we studied the field around GLEAM-X J1627 using archival European Southern Observatory and DECam Plane Survey data, as well as recent Southern African Large Telescope observations. Many sources are present close to the position of GLEAM-X J1627, but only two within the 2'' radio position uncertainty. Depending on the assumed spectral distribution, the upper limits converted to an X-ray luminosity of LX < 6.5 × 1029 erg s−1 for a blackbody with temperature kT = 0.3 keV, or LX < 9 × 1029 erg s−1 for a power law with photon index Γ = 2 (assuming a 1.3 kpc distance). Furthermore, we performed magneto-thermal simulations for neutron stars considering crust- and core-dominated field configurations. Based on our multiband limits, we conclude that (i) in the magnetar scenario, the X-ray upper limits suggest that GLEAM-X J1627 should be older than ∼1 Myr, unless it has a core-dominated magnetic field or has experienced fast cooling; (ii) in the white dwarf scenario, we can rule out most binary systems, a hot sub-dwarf, and a hot magnetic isolated white dwarf (T ≳ 10.000 K), while a cold isolated white dwarf is still compatible with our limits.N.R., F.C.Z., C.D., M.R., V.G., C.P., A.B., and E.P. are supported by the ERC Consolidator Grant "MAGNESIA" under grant agreement No. 817661, and National Spanish grant No. PGC2018-095512-BI00. F.C.Z., A.B., and V.G. are also supported by Juan de la Cierva Fellowships. C.D., M.R., and C.A.'s work has been carried out within the framework of the doctoral program in Physics of the Universitat Autónoma de Barcelona. N.H.W. is supported by an Australian Research Council Future Fellowship (project number FT190100231) funded by the Australian Government. D.d.M. acknowledges financial support from the Italian Space Agency (ASI) and National Institute for Astrophysics (INAF) under agreements ASI-INAF I/037/12/0 and ASI-INAF n.2017-14-H.0 and from INAF "Sostegno alla ricerca scientifica main streams dell'INAF," Presidential Decree 43/2018 and from INAF "SKA/CTA projects," Presidential Decree 70/2016. D.B. acknowledges support from the South African National Research Foundation. D.V. is supported by the ERC Starting Grant "IMAGINE" under grant agreement No. 948582. This work was also partially supported by the program Unidad de Excelencia Maria de Maetzu de Maeztu CEX2020-001058-M and by the PHAROS COST Action (grant No. CA16214)

Deep X-ray and radio observations of the first outburst of the young magnetar swift J1818.0−1607

Swift J1818.0−1607 is a radio-loud magnetar with a spin period of 1.36 s and a dipolar magnetic field strength of B ∼ 3 × 1014 G, which is very young compared to the Galactic pulsar population. We report here on the long-term X-ray monitoring campaign of this young magnetar using XMM-Newton, NuSTAR, and Swift from the activation of its first outburst in 2020 March until 2021 October, as well as INTEGRAL upper limits on its hard X-ray emission. The 1–10 keV magnetar spectrum is well modeled by an absorbed blackbody with a temperature of kTBB ∼ 1.1 keV and apparent reduction in the radius of the emitting region from ∼0.6 to ∼0.2 km. We also confirm the bright diffuse X-ray emission around the source extending between ∼50'' and ∼110''. A timing analysis revealed large torque variability, with an average spin-down rate $\dot{\nu }\,\sim$ −2.3 × 10−11 Hz2 that appears to decrease in magnitude over time. We also observed Swift J1818.0−1607 with the Karl G. Jansky Very Large Array on 2021 March 22. We detected the radio counterpart to Swift J1818 measuring a flux density of Sv = 4.38 ± 0.05 mJy at 3 GHz and a half-ringlike structure of bright diffuse radio emission located at ∼90'' to the west of the magnetar. We tentatively suggest that the diffuse X-ray emission is due to a dust-scattering halo and that the radio structure may be associated with the supernova remnant of this young pulsar, based on its morphology.A.Y.I.'s work has been carried out within the framework of the doctoral program in Physics of the Universitat Autònoma de Barcelona. A.Y.I, A.B., N.R., F.C.Z., E.P., R.S., S.A., V.G., C.D., and M.R. are supported by the H2020 ERC Consolidator Grant "MAGNESIA" under grant agreement No. 817661 (PI: Rea) and National Spanish grant PGC2018-095512-BI00. F.C.Z and V.G. are supported by Juan de la Cierva fellowships. A.B. acknowledge support from the Consejería de Economía, Conocimiento y Empleo del Gobierno de Canarias and the European Regional Development Fund (ERDF) under grant with reference ProID2021010132 ACCISI/FEDER, UE. T.D.R. acknowledges financial contribution from the agreement ASI-INAF n.2017-14-H.0. M.R. acknowledges financial support from the Italian Ministry for Education, University and Research through grant 2017LJ39LM "UnIAM" and the INAF Main-streams' funding grant (DP n.43/18). S.L. acknowledges financial support from the Italian Ministry of University and Research—Project Proposal CIR01_00010. This work was also partially supported by the program Unidad de Excelencia María de Maeztu CEX2020-001058-M, and by the PHAROS COST Action (No. CA16214).Peer reviewe

Assessing Trustworthy AI in times of COVID-19. Deep Learning for predicting a multi-regional score conveying the degree of lung compromise in COVID-19 patients

Abstract—The paper's main contributions are twofold: to demonstrate how to apply the general European Union’s High-Level Expert Group’s (EU HLEG) guidelines for trustworthy AI in practice for the domain of healthcare; and to investigate the research question of what does “trustworthy AI” mean at the time of the COVID-19 pandemic. To this end, we present the results of a post-hoc self-assessment to evaluate the trustworthiness of an AI system for predicting a multi-regional score conveying the degree of lung compromise in COVID-19 patients, developed and verified by an interdisciplinary team with members from academia, public hospitals, and industry in time of pandemic. The AI system aims to help radiologists to estimate and communicate the severity of damage in a patient’s lung from Chest X-rays. It has been experimentally deployed in the radiology department of the ASST Spedali Civili clinic in Brescia (Italy) since December 2020 during pandemic time. The methodology we have applied for our post-hoc assessment, called Z-Inspection®, uses socio-technical scenarios to identify ethical, technical and domain-specific issues in the use of the AI system in the context of the pandemic.</p

Circulatory Adipokines and Incretins in Adolescent Idiopathic Scoliosis: A Pilot Study

Adolescent idiopathic scoliosis (AIS) is a three-dimensional malformation of the spine of unknown cause that develops between 10 and 18 years old and affects 2&ndash;3% of adolescents, mostly girls. It has been reported that girls with AIS have a taller stature, lower body mass index (BMI), and bone mineral density (BMD) than their peers, but the causes remain unexplained. Energy metabolism discrepancies, including alterations in adipokine and incretin circulatory levels, could influence these parameters and contribute to disease pathophysiology. This pilot study aims to compare the anthropometry, BMD, and metabolic profile of 19 AIS girls to 19 age-matched healthy controls. Collected data include participants&rsquo; fasting metabolic profile, anthropometry (measurements and DXA scan), nutritional intake, and physical activity level. AIS girls (14.8 &plusmn; 1.7 years, Cobb angle 27 &plusmn; 10&deg;), compared to controls (14.8 &plusmn; 2.1 years), were leaner (BMI-for-age z-score &plusmn; SD: &minus;0.59 &plusmn; 0.81 vs. 0.09 &plusmn; 1.11, p = 0.016; fat percentage: 24.4 &plusmn; 5.9 vs. 29.2 &plusmn; 7.2%, p = 0.036), had lower BMD (total body without head z-score &plusmn; SD: &minus;0.6 &plusmn; 0.83 vs. 0.23 &plusmn; 0.98, p = 0.038; femoral neck z-score: &minus;0.54 &plusmn; 1.20 vs. 0.59 &plusmn; 1.59, p = 0.043), but their height was similar. AIS girls had higher adiponectin levels [56 (9&ndash;287) vs. 32 (7&ndash;74) &mu;g/mL, p = 0.005] and lower leptin/adiponectin ratio [0.042 (0.005&ndash;0.320) vs. 0.258 (0.024&ndash;1.053), p = 0.005]. AIS participants with a Cobb angle superior to 25&deg; had higher resistin levels compared to controls [98.2 (12.8&ndash;287.2) vs. 32.1 (6.6&ndash;73.8), p = 0.0013]. This pilot study suggests that adipokines are implicated in AIS development and/or progression, but more work is needed to confirm their role in the disease