Lipid Rafts and Clathrin Cooperate in the Internalization of PrPC in Epithelial FRT Cells

The cellular prion protein (PrP(C)) plays a key role in the pathogenesis of Transmissible Spongiform Encephalopathies in which the protein undergoes post-translational conversion to the infectious form (PrP(Sc)). Although endocytosis appears to be required for this conversion, the mechanism of PrP(C) internalization is still debated, as caveolae/raft- and clathrin-dependent processes have all been reported to be involved. METHODOLOGY/PRINCIPAL FINDINGS: We have investigated the mechanism of PrP(C) endocytosis in Fischer Rat Thyroid (FRT) cells, which lack caveolin-1 (cav-1) and caveolae, and in FRT/cav-1 cells which form functional caveolae. We show that PrP(C) internalization requires activated Cdc-42 and is sensitive to cholesterol depletion but not to cav-1 expression suggesting a role for rafts but not for caveolae in PrP(C) endocytosis. PrP(C) internalization is also affected by knock down of clathrin and by the expression of dominant negative Eps15 and Dynamin 2 mutants, indicating the involvement of a clathrin-dependent pathway. Notably, PrP(C) co-immunoprecipitates with clathrin and remains associated with detergent-insoluble microdomains during internalization thus indicating that PrP(C) can enter the cell via multiple pathways and that rafts and clathrin cooperate in its internalization. CONCLUSIONS/SIGNIFICANCE: These findings are of particular interest if we consider that the internalization route/s undertaken by PrP(C) can be crucial for the ability of different prion strains to infect and to replicate in different cell lines

Progress in low-field benchtop NMR spectroscopy in chemical and biochemical analysis

The employment of spectroscopically-resolved NMR techniques as analytical probes have previously been both prohibitively expensive and logistically challenging in view of the large sizes of high-field facilities. However, with recent advances in the miniaturisation of magnetic resonance technology, low-field, cryogen-free “benchtop” NMR instruments are seeing wider use. Indeed, these miniaturised spectrometers are utilised in areas ranging from food and agricultural analyses, through to human biofluid assays and disease monitoring. Therefore, it is both intrinsically timely and important to highlight current applications of this analytical strategy, and also provide an outlook for the future, where this approach may be applied to a wider range of analytical problems, both qualitatively and quantitatively

Dating the emergence of dairying by the first farmers of Central Europe using 14C analysis of fatty acids preserved in pottery vessels

Direct, accurate, and precise dating of archaeological pottery vessels is now achievable using a recently developed approach based on the radiocarbon dating of purified molecular components of food residues preserved in the walls of pottery vessels. The method targets fatty acids from animal fat residues, making it uniquely suited for directly dating the inception of new food commodities in prehistoric populations. Here, we report a large-scale application of the method by directly dating the introduction of dairying into Central Europe by the Linearbandkeramik (LBK) cultural group based on dairy fat residues. The radiocarbon dates (n=27) from the 54th century BC from the western and eastern expansion of the LBK suggest dairy exploitation arrived with the first settlers in the respective regions and were not gradually adopted later. This is particularly significant, as contemporaneous LBK sites showed an uneven distribution of dairy exploitation. Significantly, our findings demonstrate the power of directly dating the introduction of new food commodities, hence removing taphonomic uncertainties when assessing this indirectly based on associated cultural materials or other remainsPeer reviewe

All-electrical detection of the spin-charge conversion in nanodevices based on SrTiO3 two-dimensional electron gases

The Magnetoelectric Spin-Orbit (MESO) technology aims to bring logic into memory by combining a ferromagnet with a magnetoelectric (ME) element for information writing, and a spin-orbit (SO) element for information read-out through spin-charge conversion. Among candidate SO materials to achieve a large MESO output signal, oxide Rashba two-dimensional electron gases (2DEGs) have shown very large spin-charge conversion efficiencies, albeit mostly in spin-pumping experiments. Here, we report all-electrical spin-injection and spin-charge conversion experiments in nanoscale devices harnessing the inverse Edelstein effect of SrTiO3 2DEGs. We have designed, patterned and fabricated nanodevices in which a spin current injected from a cobalt layer into the 2DEG is converted into a charge current. We optimized the spin-charge conversion signal by applying back-gate voltages, and studied its temperature evolution. We further disentangled the inverse Edelstein contribution from spurious effects such as the planar Hall effect, the anomalous Hall effect or the anisotropic magnetoresistance. The combination of non-volatility and high energy efficiency of these devices could potentially lead to new technology paradigms for beyond-CMOS computing architectures

La longue séquence de Marchésieux: reconstitution de paléoenvironnements marins durant le premier cycle glaciaire de l'hémisphère nord

International audienceA l'échelle des cinq derniers millions d'années, les enregistrements paléoclimatiques restitués par les sédiments océaniques illustrent le contrôle des paramètres orbitaux (précession, obliquité et excentricité) sur le climat global. La tendance au refroidissement qui accompagne la fin du Néogène est ainsi marquée, vers -2,4 Ma, par le développement rapide des calottes de glace de l'hémisphère nord (Shackleton et al., 1984). Alors que l'histoire du climat global restituée par les enregistrements océaniques est sans cesse précisée, les données concernant l'évolution des environnements continentaux et côtiers au cours de ces changements restent essentiellement fragmentaires. Le forage effectué à Marchésieux (Manche; Normandie) a permis de réaliser une étude pluridisciplinaire de la signature de ce premier cycle glaciaire de l'hémisphère nord (Prétiglien). Les premiers résultats stratigraphiques, paléoenvironnementaux (eustatisme, température, paléobathymétrie) sont présentés

Herpes simplex virus encephalitis in a patient with complete TLR3 deficiency: TLR3 is otherwise redundant in protective immunity

A new autosomal recessive form of complete TLR3 deficiency reveals that human TLR3 is nonredundant in immunity against herpes simplex virus 1 in the central nervous system (CNS) but redundant in host defense against viruses outside the CNS

A biobank of pediatric patient-derived-xenograft models in cancer precision medicine trial MAPPYACTS for relapsed and refractory tumors

Pediatric patients with recurrent and refractory cancers are in most need for new treatments. This study developed patient-derived-xenograft (PDX) models within the European MAPPYACTS cancer precision medicine trial (NCT02613962). To date, 131 PDX models were established following heterotopical and/or orthotopical implantation in immunocompromised mice: 76 sarcomas, 25 other solid tumors, 12 central nervous system tumors, 15 acute leukemias, and 3 lymphomas. PDX establishment rate was 43%. Histology, whole exome and RNA sequencing revealed a high concordance with the primary patient's tumor profile, human leukocyte-antigen characteristics and specific metabolic pathway signatures. A detailed patient molecular characterization, including specific mutations prioritized in the clinical molecular tumor boards are provided. Ninety models were shared with the IMI2 ITCC Pediatric Preclinical Proof-of-concept Platform (IMI2 ITCC-P4) for further exploitation. This PDX biobank of unique recurrent childhood cancers provides an essential support for basic and translational research and treatments development in advanced pediatric malignancies

More rapid blood interferon α2 decline in fatal versus surviving COVID-19 patients

BackgroundThe clinical outcome of COVID-19 pneumonia is highly variable. Few biological predictive factors have been identified. Genetic and immunological studies suggest that type 1 interferons (IFN) are essential to control SARS-CoV-2 infection.ObjectiveTo study the link between change in blood IFN-α2 level and plasma SARS-Cov2 viral load over time and subsequent death in patients with severe and critical COVID-19.MethodsOne hundred and forty patients from the CORIMUNO-19 cohort hospitalized with severe or critical COVID-19 pneumonia, all requiring oxygen or ventilation, were prospectively studied. Blood IFN-α2 was evaluated using the Single Molecule Array technology. Anti-IFN-α2 auto-Abs were determined with a reporter luciferase activity. Plasma SARS-Cov2 viral load was measured using droplet digital PCR targeting the Nucleocapsid gene of the SARS-CoV-2 positive-strand RNA genome.ResultsAlthough the percentage of plasmacytoid dendritic cells was low, the blood IFN-α2 level was higher in patients than in healthy controls and was correlated to SARS-CoV-2 plasma viral load at entry. Neutralizing anti-IFN-α2 auto-antibodies were detected in 5% of patients, associated with a lower baseline level of blood IFN-α2. A longitudinal analysis found that a more rapid decline of blood IFN-α2 was observed in fatal versus surviving patients: mortality HR=3.15 (95% CI 1.14–8.66) in rapid versus slow decliners. Likewise, a high level of plasma SARS-CoV-2 RNA was associated with death risk in patients with severe COVID-19.ConclusionThese findings could suggest an interest in evaluating type 1 IFN treatment in patients with severe COVID-19 and type 1 IFN decline, eventually combined with anti-inflammatory drugs.Clinical trial registrationhttps://clinicaltrials.gov, identifiers NCT04324073, NCT04331808, NCT04341584

Accurate compound-specific 14C dating of archaeological pottery vessels

Pottery is one of the most commonly recovered artefacts from archaeological sites. Despite more than a century of relative dating based on typology and seriation, accurate dating of pottery by the radiocarbon method has proven extremely challenging due to the limited survival of organic temper and unreliability of visible residues. We report here a new method of dating directly archaeological pottery based on accelerator mass spectrometry (AMS) analysis of 14C in absorbed food residues: palmitic (C16:0) and stearic (C18:0) fatty acids purified by preparative gas chromatography (pcGC). We present the first accurate compound-specific radiocarbon determinations of lipids extracted from pottery vessels, which were rigorously evaluated by comparison with dendrochronological dates and inclusion in site and regional chronologies containing suites of radiocarbon dates on other materials . Critically, the compound-specific dates from each of the C16:0 and C18:0 fatty acids in pottery vessels provide an internal quality control of the results and, are entirely compatible with dates for other commonly dated materials. Accurate radiocarbon dating of pottery vessels can reveal: (i) the period of use of pottery; (ii) the antiquity of organic residues including when specific foodstuffs were exploited; (iii) sites chronologies in the absence of traditionally datable materials and (iv) direct verification of pottery typochronologies. As exemplars, the method was applied to the dating of dairy and carcass product exploitation in Neolithic vessels, from Britain, Anatolia, central and western Europe, and Saharan Africa.Additional co-authors: Christian Jeunesse, Marta Krueger, Arkadiusz Marciniak, Steve Minnitt, Rocco Rotunno, Pieter van de Velde, Ivo van Wijk, Jonathan Cotton, Andy Daykin, Richard P Evershe