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NJFVID+SPSMTSYBBold Type 1C Custom yes yes yes 284 0 WarnockPro-Bold Type 1C WinAnsi yes no yes 376 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-10-08 03:54:45 CEST RepresentationInformation: ANC_a2023v11p143.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-10-07 12:21:17 CEST Size: 4114543 Format: PDF Version: 1.4 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 1360 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: false CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseOutlines Language: EN Outlines: Item: Title: Understanding the key features of the spontaneous formation of bona fide prions through a novel methodology that enables their swift and consistent generation Children: Item: Title: Abstract Item: Title: Introduction Item: Title: Material and methods Children: Item: Title: Production and purification of recombinant PrP Item: Title: Preparation of PMSA substrates Item: Title: PMSA reaction for spontaneous misfolding of recombinant PrP and its in vitro propagation Item: Title: Loading of prions in zirconium silicate beads for long term storage and use as a seed Item: Title: Misfolded recombinant PrP detection Item: Title: PMCA for the assessment of the potential infectivity of recombinant misfolded PrP Children: Item: Title: Preparation of PMCA substrate Item: Title: Preparation of recombinant seeds Item: Title: Brain-PMCA Item: Title: Bioassay in TgVole 1 × mouse model Children: Item: Title: Preparation of inocula Item: Title: Animal inoculation Item: Title: Detection of PrPSc in brain and PMCA samples by Western blotting Item: Title: Anatomopathological analysis and immunohistochemistry Item: Title: Cryo-electron microscopy Item: Title: Scanning electron microscopy (SEM) Item: Title: Mass spectrometry Item: Title: Siliconization of glass beads for PMSA Item: Title: PMSA in glass tube versus plastic tube with and without glass beads Item: Title: Immobilization of glass beads in plastic resin for PMSA Item: Title: Evaluation of rec-PrP adsorption to glass beads Item: Title: Recombinant prion propagation in PMSA using prion-coated glass spheres as seed Item: Title: Determination of the dynamics of spontaneous misfolding and prion propagation in PMSA Item: Title: Results Children: Item: Title: A highly efficient technique to propagate bona fide prions reveals spontaneous stochastic protein misfolding events Item: Title: Only a certain type of beads efficiently drives spontaneous recombinant PrP misfolding Item: Title: A highly reproducible and truly spontaneous protein misfolding event Item: Title: Recombinant PrP spontaneously misfolded by PMSA exhibits the key biochemical hallmarks of bona fide prions Item: Title: Distinct electrophoretic migration profiles point towards the spontaneous formation of different conformers Item: Title: Spontaneous misfolding in PMSA consistently generates bona fide prions that are infectious in vivo and exhibit distinct strain features Item: Title: Recombinant prions generated spontaneously in PMSA show high specific infectivity comparable to those of brain-derived prions Item: Title: Electron microscopy and mass spectrometry analysis of Ust02 recombinant prion strain confirms its fibrillary structure and gain insights into the structural arrangement of recombinant prions Item: Title: The extension of the glass surface is critical for the spontaneous misfolding Item: Title: The highly efficient spontaneous misfolding also requires the movement or interaction of the glass beads Item: Title: The interaction between recombinant PrP and the surface of glass beads is essential for inducing spontaneous misfolding Item: Title: Spontaneously formed rec-PrPres is strongly associated to glass beads, which can act as seeds in vitro and are highly infectious in vivo Item: Title: Exploring the dynamics of spontaneous misfolding and prion propagation in PMSA Item: Title: Discussion Item: Title: Anchor 43 Item: Title: Acknowledgements Item: Title: References Info: Title: Understanding the key features of the spontaneous formation of bona fide prions through a novel methodology that enables their swift and consistent generation Author: Hasier Eraña Subject: Acta Neuropathologica Communications, https://doi.org/10.1186/s40478-023-01640-8 Keywords: Prion; Transmissible spongiform encephalopathies; Spontaneous misfolding; Strain Creator: Springer Producer: Acrobat Distiller 10.1.8 (Windows); modified using iText® 5.3.5 ©2000-2012 1T3XT BVBA (SPRINGER SBM; licensed version) CreationDate: Tue Sep 05 20:57:27 CEST 2023 ModDate: Mon Nov 20 06:19:18 CET 2023 ID: 0x14888149c81d69d716eb8d0406ee4e4a, 0x006afb3696d99c4c8bb6c30b6f156a64 Filters: FilterPipeline: FlateDecode 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Díaz-Domínguez Jorge M. Charco Enric Vidal Ezequiel González-Miranda Miguel A. Pérez-Castro Patricia Piñeiro Rafael López-Moreno Cristina Sampedro-Torres-Quevedo Leire Fernández-Veiga Juan Tasis-Galarza Nuria L. Lorenzo Aileen Santini-Santiago Melisa Lázaro Sandra García-Martínez Nuno Gonçalves-Anjo Maitena San-Juan-Ansoleaga Josu Galarza-Ahumada Eva Fernández-Muñoz Samanta Giler Mikel Valle Glenn C. Telling Mariví Geijó Jesús R. 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