The involvement of dityrosine crosslinking in α-synuclein assembly and deposition in Lewy Bodies in Parkinson’s disease

Parkinson’s disease (PD) is characterized by intracellular, insoluble Lewy bodies composed of highly stable α-synuclein (α-syn) amyloid fibrils. α-synuclein is an intrinsically disordered protein that has the capacity to assemble to form β-sheet rich fibrils. Oxidiative stress and metal rich environments have been implicated in triggering assembly. Here, we have explored the composition of Lewy bodies in post-mortem tissue using electron microscopy and immunogold labeling and revealed dityrosine crosslinks in Lewy bodies in brain tissue from PD patients. In vitro, we show that dityrosine cross-links in α-syn are formed by covalent ortho-ortho coupling of two tyrosine residues under conditions of oxidative stress by fluorescence and confirmed using mass-spectrometry. A covalently cross-linked dimer isolated by SDS-PAGE and mass analysis showed that dityrosine dimer was formed via the coupling of Y39-Y39 to give a homo dimer peptide that may play a key role in formation of oligomeric and seeds for fibril formation. Atomic force microscopy analysis reveals that the covalent dityrosine contributes to the stabilization of α-syn assemblies. Thus, the presence of oxidative stress induced dityrosine could play an important role in assembly and toxicity of α-syn in PD

Blood flow and coherent vortices in the normal and aneurysmatic aortas: a fluid dynamical approach to intra-luminal thrombus formation

Abdominal aortic aneurysms (AAAs) are frequently characterized by the development of an intra-luminal thrombus (ILT), which is known to have multiple biochemical and biomechanical implications. Development of the ILT is not well understood, and shear–stress-triggered activation of platelets could be the first step in its evolution. Vortical structures (VSs) in the flow affect platelet dynamics, which motivated the present study of a possible correlation between VS and ILT formation in AAAs. VSs educed by the λ2-method using computational fluid dynamics simulations of the backward-facing step problem, normal aorta, fusiform AAA and saccular AAA were investigated. Patient-specific luminal geometries were reconstructed from computed tomography scans, and Newtonian and Carreau–Yasuda models were used to capture salient rheological features of blood flow. Particularly in complex flow domains, results depended on the constitutive model. VSs developed all along the normal aorta, showing that a clear correlation between VSs and high wall shear stress (WSS) existed, and that VSs started to break up during late systole. In contrast, in the fusiform AAA, large VSs developed at sites of tortuous geometry and high WSS, occupying the entire lumen, and lasting over the entire cardiac cycle. Downward motion of VSs in the AAA was in the range of a few centimetres per cardiac cycle, and with a VS burst at that location, the release (from VSs) of shear-stress-activated platelets and their deposition to the wall was within the lower part of the diseased artery, i.e. where the thickest ILT layer is typically observed. In the saccular AAA, only one VS was found near the healthy portion of the aorta, while in the aneurysmatic bulge, no VSs occurred. We present a fluid-dynamics-motivated mechanism for platelet activation, convection and deposition in AAAs that has the potential of improving our current understanding of the pathophysiology of fluid-driven ILT growth

Tau (297‐391) forms filaments that structurally mimic the core of paired helical filaments in Alzheimer’s disease brain

The constituent paired helical filaments (PHFs) in neurofibrillary tangles are insoluble intracellular deposits central to the development of Alzheimer’s disease (AD) and other tauopathies. Full‐length tau requires the addition of anionic cofactors such as heparin to enhance assembly. We have shown that a fragment from the proteolytically stable core of the PHF, tau 297‐391 known as ‘dGAE’, spontaneously forms cross‐β‐containing PHFs and straight filaments under physiological conditions. Here, we have analysed and compared the structures of the filaments formed by dGAE in vitro with those deposited in the brains of individuals diagnosed with AD. We show that dGAE forms PHFs that share a macromolecular structure similar to those found in brain tissue. Thus, dGAEs may serve as a model system for studying core domain assembly and for screening for inhibitors of tau aggregation

Análisis de la aplicabilidad del modelo de Bohr acerca de la predicción de las líneas de emisión del átomo de helio en el marco del dictado de Física Moderna en la Facultad de Ingeniería de la UNLP

El dictado de la materia Física IIIA de las carreras de Ingeniería Electrónica (IE1), Electricista (IE2) y Química (IQ) dictadas en la Facultad de Ingeniería (FI) de la Universidad Nacional de La Plata (UNLP) abarca dos módulos bimestrales. En el segundo de ellos se abordan temas de Física Moderna y entre las actividades de laboratorio establecidas, los alumnos deben realizar un experimento destinado a determinar las líneas características de emisión de una lámpara de gas de He a baja presión utilizando un espectrómetro portátil. En el presente trabajo se describe el instrumental utilizado, la metodología experimental adoptada, se presentan las mediciones realizadas de las líneas características emitidas por el átomo He (HeI) y se las compara con las líneas publicadas en la literatura. A partir del uso del modelo de Bohr para el caso del átomo de He+1 simplemente ionizado (HeII), se estiman las longitudes de onda emitidas por el ión HeII y se las compara con las líneas características del HeII publicadas. También en el marco del modelo de Bohr, se calculan las longitudes de onda emitidas por el átomo HeI despreciando la repulsión entre los electrones y se las compara con las líneas observadas en el laboratorio. Finalmente, se analiza la aplicabilidad del modelo de Bohr en la descripción de los sistemas HeI y HeII.The Physic IIIA course of the Electronic, Electric and Chemical Engineering degrees of La Plata University includes two modules of two months each. The second teaching module includes Modern Physic topics. In special, in the laboratory activities, students carry out an experiment to determine the characteristic emission lines of an He lamp using a didactic spectrometer. In the present work, we describe the approach used in the laboratory and the experimental methodology. We present the emission lines measured and we compare these measurements with the He emission lines reported in the literature. From the Bohr model, we estimate the emitted wave length for the case of the He+1 ion (The He atom without an electron) and we compare these wave lengths with the lines published. In this sense, we also calculate the emitted lines of the He atom in the frame of the Bohr model and we compare these calculations with the lines measured in the laboratory. Finally, we analyze the potential application of the Bohr model in the description of the He atom and of the He+1 ion.Trabajo presentado en la 96A Reunión Nacional de Física y II Reunión Conjunta Sociedad Uruguaya de Física - AsociacióN Física Argentin

Conservation research in times of COVID-19 - the rescue of the northern white rhino

COVID-19 has changed the world at unprecedented pace. The measures imposed by governments across the globe for containing the pandemic have severely affected all facets of economy and society, including scientific progress. Сonservation research has not been exempt from these negative effects, which we here summarize for the BioRescue project, aiming at saving the northern white rhinoceros (Ceratotherium simum cottoni), an important Central African keystone species, of which only two female individuals are left. The development of advanced assisted reproduction and stem-cell technologies to achieve this goal involves experts across five continents. Maintaining international collaborations under conditions of national shut-down and travel restrictions poses major challenges. The associated ethical implications and consequences are particularly troublesome when it comes to research directed at protecting biological diversity – all the more in the light of increasing evidence that biodiversity and intact ecological habitats might limit the spread of novel pathogens

Paired helical filament-forming region of tau (297–391) influences endogenous tau protein and accumulates in acidic compartments in human neuronal cells

Assembly of tau protein into paired helical filaments and straight filaments is a key feature of Alzheimer's disease. Aggregation of tau has been implicated in neurodegeneration, cellular toxicity and the propagation, which accompanies disease progression. We have reported previously that a region of tau (297–391), referred to as dGAE, assembles spontaneously in physiological conditions to form paired helical filament-like fibres in vitro in the absence of additives such as heparin. This provides a valuable tool with which to explore the effects of tau in cell culture. Here we have studied the cellular uptake of soluble oligomeric and fibrillar forms of dGAE and examined the downstream consequences of tau internalisation into differentiated SH-SY5Y neuroblastoma cells using fluorescence and electron microscopy alongside structural and biochemical analyses. The assembled dGAE shows more acute cytotoxicity than the soluble, non-aggregated form. Conversely, the soluble form is much more readily internalised and, once within the cell, is able to associate with endogenous tau resulting in increased phosphorylation and aggregation of endogenous tau, which accumulates in lysosomal/endosomal compartments. It appears that soluble oligomeric forms are able to propagate tau pathology without being acutely toxic. The model system we have developed now permits the molecular mechanisms of propagation of tau pathology to be studied in vitro in a more physiological manner with a view to development of novel therapeutic approaches

An Integrated Fluid-Chemical Model Toward Modeling the Formation of Intra-Luminal Thrombus in Abdominal Aortic Aneurysms

Abdominal Aortic Aneurysms (AAAs) are frequently characterized by the presence of an Intra-Luminal Thrombus (ILT) known to influence their evolution biochemically and biomechanically. The ILT progression mechanism is still unclear and little is known regarding the impact of the chemical species transported by blood flow on this mechanism. Chemical agonists and antagonists of platelets activation, aggregation, and adhesion and the proteins involved in the coagulation cascade (CC) may play an important role in ILT development. Starting from this assumption, the evolution of chemical species involved in the CC, their relation to coherent vortical structures (VSs) and their possible effect on ILT evolution have been studied. To this end a fluid-chemical model that simulates the CC through a series of convection-diffusion-reaction (CDR) equations has been developed. The model involves plasma-phase and surface-bound enzymes and zymogens, and includes both plasma-phase and membrane-phase reactions. Blood is modeled as a non-Newtonian incompressible fluid. VSs convect thrombin in the domain and lead to the high concentration observed in the distal portion of the AAA. This finding is in line with the clinical observations showing that the thickest ILT is usually seen in the distal AAA region. The proposed model, due to its ability to couple the fluid and chemical domains, provides an integrated mechanochemical picture that potentially could help unveil mechanisms of ILT formation and development

Buried waveguides in Nd:YLF crystals obtained by femtosecond laser writing under double line approach

In this paper, we present buried waveguides fabricated by fs laser writing in Nd3+ doped YLF crystal under double line approach (Miura et al. in Appl. Phys. Lett. 71:3329?3331, 1997). The waveguides were made by focusing two consecutive optical breakdown tracks (OBT) separated by about 20 μm. To make the optimal OBT, we focused the fs-laser pulses 200 μm below surface at intensities above the OB threshold for the material and controlled the writing speed. The guiding structures were fabricated by using a Chirped Pulse Amplification (CPA) femtosecond (fs) laser system. We chose the optimal writing parameters in order to obtain suitable waveguides, using around 3 μJ energy and writing speed from 15 to 50 μm/s. After optically exploring the waveguides by end-fire coupling, the guiding structures showed good optical performance. Guiding index profiles were retrieved from modal analysis by using BeamProp (RSoft) commercial software. This spatial distribution of the index increment, taking into account a lower refractive barrier on the OBT region plus the compressed region between the tracks, was obtained correctly fitting profiles modes. Finally, optical spectroscopy measurements were also performed in the waveguides. The results showed that the luminescence properties of Nd3+ ions are preserved in the waveguides compared with the values obtained for bulk.Fil: Biasetti, Demian Arístide. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Investigaciones Opticas (i); ArgentinaFil: Neyra, Enrique Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Investigaciones Opticas (i); ArgentinaFil: Vázquez de Aldana, J. R.. Universidad de Salamanca; EspañaFil: Roso, L.. Universidad de Salamanca; EspañaFil: Torchia, Gustavo Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Investigaciones Opticas (i); Argentin