Non-contact, single-sided access ultrasonic guided waves for the assessment of materials mechanical properties

Abstract: Research about material characterization without contact has been carried out by many authors using immersion or laser-based ultrasonic techniques. Immersion techniques however imply that the material is not water sensitive and that the sample fits within the immersion tank. Therefore, it is important to develop a characterization process that is suitable for all types of materials, and ideally not requiring access to both sides of the tested specimen, as this is often not possible in industrial context.Résumé de la communication présentée lors du congrès international tenu conjointement par Canadian Society for Mechanical Engineering (CSME) et Computational Fluid Dynamics Society of Canada (CFD Canada), à l’Université de Sherbrooke (Québec), du 28 au 31 mai 2023

Phosphorylation and O-GlcNAcylation of the PHF-1 Epitope of Tau Protein Induce Local Conformational Changes of the C-Terminus and Modulate Tau Self-Assembly Into Fibrillar Aggregates

Phosphorylation of the neuronal microtubule-associated Tau protein plays a critical role in the aggregation process leading to the formation of insoluble intraneuronal fibrils within Alzheimer’s disease (AD) brains. In recent years, other posttranslational modifications (PTMs) have been highlighted in the regulation of Tau (dys)functions. Among these PTMs, the O-β-linked N-acetylglucosaminylation (O-GlcNAcylation) modulates Tau phosphorylation and aggregation. We here focus on the role of the PHF-1 phospho-epitope of Tau C-terminal domain that is hyperphosphorylated in AD (at pS396/pS404) and encompasses S400 as the major O-GlcNAc site of Tau while two additional O-GlcNAc sites were found in the extreme C-terminus at S412 and S413. Using high resolution NMR spectroscopy, we showed that the O-GlcNAc glycosylation reduces phosphorylation of PHF-1 epitope by GSK3β alone or after priming by CDK2/cyclin A. Furthermore, investigations of the impact of PTMs on local conformation performed in small peptides highlight the role of S404 phosphorylation in inducing helical propensity in the region downstream pS404 that is exacerbated by other phosphorylations of PHF-1 epitope at S396 and S400, or O-GlcNAcylation of S400. Finally, the role of phosphorylation and O-GlcNAcylation of PHF-1 epitope was probed in in-vitro fibrillization assays in which O-GlcNAcylation slows down the rate of fibrillar assembly while GSK3β phosphorylation stimulates aggregation counteracting the effect of glycosylation.Peer Reviewe

Role of post-translational modifications in the aggregation process of the Tau protein

L’analyse histologique de cerveaux des patients atteints par la maladie Alzheimer met en évidence deux lésions moléculaires majeures : les plaques amyloïdes extraneuronales et les structures fibrillaires intra-neuronales. Alors que les premières sont constituées par l’agrégation du peptide Aβ, les fibres intra-neuronales, aussi appelées PHFs, sont essentiellement formées par l’agrégation de la protéine Tau hyperphosphorylée. De nombreuses études ont démontré une corrélation positive entre l’hyperphosphorylation de la protéine Tau, la progression de la dégénérescence neurofibrillaire de manière spatio-temporelle dans le cerveau et le stade de la maladie mais, à ce jour, le mécanisme moléculaire liant la phosphorylation et l’agrégation pathologique reste inconnu. De plus, par l’absence de structure tridimensionnelle, la protéine Tau est une cible privilégiée d’un grand nombre d’enzymes telles que les kinases, acétyl- et glycosyl-transférases... impliquées dans la modification post-traductionnelle (MPT) de la protéine. Le rôle de ces autres modifications dans le processus agrégatif n’est pas encore clairement établi et mérite d’être étudié. De plus, l’identification de ces différentes modifications nous a amené à nous interroger sur le dialogue potentiel avec la phosphorylation et leur impact sur l’agrégation de la protéine Tau. Dans ce contexte, par différentes approches in vitro, nous avons étudié le rôle de trois modifications clés de la protéine Tau : la phosphorylation (Ser/Thr), l’acétylation (Lys) et l’O-GlcNAcylation (Ser).The brains of patients with Alzheimer’s disease (AD) are characterized at the molecular level by two different lesions: extraneuronal amyloid plaques and intraneuronal tangles. Both contain fibrillary structures: the former is composed of insoluble Aβ peptides and the tangles contain paired helical filaments (PHFs) made of Tau proteins as the main contituent.It has been established that Tau is is found in an hyperphosphorylated state in PHFs. Numerous studies have demonstrated a positive correlation between hyperphosphorylation of Tau, progression of neurofibrillary degeneration through the brain and stages of the disease but the molecular mechanisms underlying Tau hyperphosphorylation and aggregation in AD are currently unknown. Moreover, due to its intrinsically disordered nature, Tau displays an increased accessibility for lots of modifying enzymes such as kinases, acetyl- and glycosyl-transferases…which are responsible for Tau post-translational modifications (PTMs). The role of these modifications in the pathological aggregation process of Tau protein remains unclear and has to be elucidated. Moreover, identification of these PTMs raises the question of the potential crosstalk with phosphorylation on the pathological aggregation of Tau. In this context, by using different in vitro approaches, we have studied the role of three major modifications of Tau: phosphorylation (Ser/Thr), acetylation (Lys) and O-GlcNAcylation (Ser)

Characterization of viscoelastic moduli and thickness of isotropic, viscoelastic plates using multi-modal Lamb waves

This paper presents an approach exploiting the sensitivity of Lamb waves for characterizing the viscoelastic moduli and thickness of plates. The analytical sensitivity functions are first derived in the case of an isotropic plate and are integrated into an iterative inverse problem to optimize its viscoelastic moduli and thickness based on a zero-finding approach (Gauss–Newton algorithm for a multivariable problem). This method is validated numerically for a viscoelastic plate and shows high accuracy and low computational cost when compared to existing methods. Experimental validation demonstrates the ability of the algorithm to assess simultaneously the viscoelastic moduli and the thickness of isotropic plate-like structures

L'interaction entre médecins et malades productrice d'inégalités sociales de santé ? Le cas de l'obésité.

300 p

NMR meets Tau: insights into its function and pathology

In this review, we focus on what we have learned from Nuclear Magnetic Resonance (NMR) studies on the neuronal microtubule-associated protein Tau. We consider both the mechanistic details of Tau: the tubulin relationship and its aggregation process. Phosphorylation of Tau is intimately linked to both aspects. NMR spectroscopy has depicted accurate phosphorylation patterns by different kinases, and its non-destructive character has allowed functional assays with the same samples. Finally, we will discuss other post-translational modifications of Tau and its interaction with other cellular factors in relationship to its (dys)function

Native Top-Down Mass Spectrometry and Ion Mobility Spectrometry of the Interaction of Tau Protein with a Molecular Tweezer Assembly Modulator.

Native top-down mass spectrometry (MS) and ion mobility spectrometry (IMS) were applied to characterize the interaction of a molecular tweezer assembly modulator, CLR01, with tau, a protein believed to be involved in a number of neurodegenerative disorders, including Alzheimer's disease. The tweezer CLR01 has been shown to inhibit aggregation of amyloidogenic polypeptides without toxic side effects. ESI-MS spectra for different forms of tau protein (full-length, fragments, phosphorylated, etc.) in the presence of CLR01 indicate a primary binding stoichiometry of 1:1. The relatively high charging of the protein measured from non-denaturing solutions is typical of intrinsically disordered proteins, such as tau. Top-down mass spectrometry using electron capture dissociation (ECD) is a tool used to determine not only the sites of post-translational modifications but also the binding site(s) of non-covalent interacting ligands to biomolecules. The intact protein and the protein-modulator complex were subjected to ECD-MS to obtain sequence information, map phosphorylation sites, and pinpoint the sites of inhibitor binding. The ESI-MS study of intact tau proteins indicates that top-down MS is amenable to the study of various tau isoforms and their post-translational modifications (PTMs). The ECD-MS data point to a CLR01 binding site in the microtubule-binding region of tau, spanning residues K294-K331, which includes a six-residue nucleating segment PHF6 (VQIVYK) implicated in aggregation. Furthermore, ion mobility experiments on the tau fragment in the presence of CLR01 and phosphorylated tau reveal a shift towards a more compact structure. The mass spectrometry study suggests a picture for the molecular mechanism of the modulation of protein-protein interactions in tau by CLR01. Graphical Abstract ᅟ

Native Top-Down Mass Spectrometry and Ion Mobility Spectrometry of the Interaction of Tau Protein with a Molecular Tweezer Assembly Modulator

Native top-down mass spectrometry (MS) and ion mobility spectrometry (IMS) were applied to characterize the interaction of a molecular tweezer assembly modulator, CLR01, with tau, a protein believed to be involved in a number of neurodegenerative disorders, including Alzheimer's disease. The tweezer CLR01 has been shown to inhibit aggregation of amyloidogenic polypeptides without toxic side effects. ESI-MS spectra for different forms of tau protein (full-length, fragments, phosphorylated, etc.) in the presence of CLR01 indicate a primary binding stoichiometry of 1:1. The relatively high charging of the protein measured from non-denaturing solutions is typical of intrinsically disordered proteins, such as tau. Top-down mass spectrometry using electron capture dissociation (ECD) is a tool used to determine not only the sites of post-translational modifications but also the binding site(s) of non-covalent interacting ligands to biomolecules. The intact protein and the protein-modulator complex were subjected to ECD-MS to obtain sequence information, map phosphorylation sites, and pinpoint the sites of inhibitor binding. The ESI-MS study of intact tau proteins indicates that top-down MS is amenable to the study of various tau isoforms and their post-translational modifications (PTMs). The ECD-MS data point to a CLR01 binding site in the microtubule-binding region of tau, spanning residues K294-K331, which includes a six-residue nucleating segment PHF6 (VQIVYK) implicated in aggregation. Furthermore, ion mobility experiments on the tau fragment in the presence of CLR01 and phosphorylated tau reveal a shift towards a more compact structure. The mass spectrometry study suggests a picture for the molecular mechanism of the modulation of protein-protein interactions in tau by CLR01. Graphical Abstract ᅟ

Human glioblastoma-initiating cells invade specifically the subventricular zones and olfactory bulbs of mice after striatal injection.

This study reports the subsequent isolation of human glioblatoma cells able to initiate experimental brain tumors, specifically and repeatedly found in the subventricular zones and olfactory bulbs following xenograft in the caudate putamen of immunodeficient mice.In patients with glioblastoma multiforme, recurrence is the rule despite continuous advances in surgery, radiotherapy and chemotherapy. Within these malignant gliomas, glioblastoma stem cells or initiating cells have been recently described and they were shown to be specifically involved in experimental tumorigenesis. In this study, we show that some human glioblastoma cells injected into the striatum of immunodeficient nude mice exhibit a tropism for the subventricular zones. There and similarily to neurogenic stem cells, these subventricular glioblastoma cells were then able to migrate towards the olfactory bulbs. Finally, the glioblastoma cells isolated from the adult mouse subventricular zones and olfactory bulbs display high tumorigenicity when secondary injected in a new mouse brain. Together, these data suggest that neurogenic zones could be a reservoir for particular cancer-initiating cells

Major Differences between the Self-Assembly and Seeding Behavior of Heparin-Induced and in Vitro Phosphorylated Tau and Their Modulation by Potential Inhibitors.

Self-assembly of the microtubule-associated protein tau into neurotoxic oligomers, fibrils, and paired helical filaments, and cell-to-cell spreading of these pathological tau species are critical processes underlying the pathogenesis of Alzheimers disease and other tauopathies. Modulating the self-assembly process and inhibiting formation and spreading of such toxic species are promising strategies for therapy development. A challenge in investigating tau self-assembly in vitro is that, unlike most amyloidogenic proteins, tau does not aggregate in the absence of posttranslational modifications (PTM), aggregation inducers, or preformed seeds. The most common induction method is addition of polyanions, such as heparin; yet, this artificial system may not represent adequately tau self-assembly in vivo, which is driven by aberrant phosphorylation and other PTMs, potentially leading to in vitro data that do not reflect the behavior of tau and its interaction with modulators in vivo. To tackle these challenges, methods for in vitro phosphorylation of tau to produce aggregation-competent forms recently have been introduced ( Despres et al. ( 2017 ) Proc. Natl. Acad. Sci. U.S.A. , 114 , 9080 - 9085 ). However, the oligomerization, seeding, and interaction with assembly modulators of the different forms of tau have not been studied to date. To address these knowledge gaps, we compared here side-by-side the self-assembly and seeding activity of heparin-induced tau with two forms of in vitro phosphorylated tau and tested how the molecular tweezer CLR01, a negatively charged compound, affected these processes. Tau was phosphorylated by incubation either with activated extracellular signal-regulated kinase 2 or with a whole rat brain extract. Seeding activity was measured using a fluorescence-resonance energy transfer-based biosensor-cell method. We also used solution-state NMR to investigate the binding sites of CLR01 on tau and how they were impacted by phosphorylation. Our systematic structure-activity relationship study demonstrates that heparin-induced tau behaves differently from in vitro phosphorylated tau. The aggregation rates of the different forms are distinct as is the intracellular localization of the induced aggregates, which resemble brain-derived tau strains suggesting that heparin-induced tau and in vitro phosphorylated tau have different conformations, properties, and activities. CLR01 inhibits aggregation and seeding of both heparin-induced and in vitro phosphorylated tau dose-dependently, although heparin induction interferes with the interaction between CLR01 and tau