CD73 represses pro-inflammatory responses in human endothelial cells

<p>Abstract</p> <p>Background</p> <p>CD73 is a 5'-ectonucleotidase that produces extracellular adenosine, which then acts on G protein-coupled purigenic receptors to induce cellular responses. CD73 has been reported to regulate expression of pro-inflammatory molecules in mouse endothelium. Our aim is to determine the function of CD73 in human endothelial cells.</p> <p>Methods</p> <p>We used RNAi to deplete CD73 levels in human umbilical cord endothelial cells (HUVECs).</p> <p>Results</p> <p>CD73 depletion resulted in a strong reduction in adenosine production, indicating that CD73 is the major source of extracellular adenosine in HUVECs. We find that CD73 depletion induces a similar response to pro-inflammatory stimuli such as the cytokine TNF-α. In CD73-depleted cells, surface levels of the leukocyte adhesion molecules ICAM-1, VCAM-1 and E-selectin increase. This correlates with increased translocation of the transcription factor NF-kB to the nucleus, which is known to regulate ICAM-1, VCAM-1 and E-selectin expression in response to TNF-α. Adhesion of monocytic cells to endothelial cells is enhanced. In addition, CD73-depleted cells become elongated, have higher levels of stress fibres and increased endothelial permeability, resembling known responses to TNF-α.</p> <p>Conclusions</p> <p>These results indicate that CD73 normally suppresses pro-inflammatory responses in human endothelial cells.</p

Argiotalus, fils de Smertulitanus, cavalier namnète à Worms (Allemagne) sous Tibère

Un moulage de la stèle figurée d’Argiotalus, fils de Smertulitanus, Namnète, cavalier de l’ala Indiana Gallorum, mort à Worms sous Tibère, est entré au musée départemental Dobrée, à Nantes. La relecture du monument funéraire du plus ancien Namnète connu est l’occasion de le situer parmi les stèles de cavaliers auxiliaires de Worms et de Mayence, dans le contexte historique et militaire d’une Gaule en voie de romanisation, mais encore agitée de révoltes comme celle, en 21, des Andes et des Turons, puis du Trévire Iulius Florus et de l’Éduen Iulius Sacrovir.A cast from the figurative stela of Argiotalus, son of Smertulitanus, a Namnet cavalryman of the Ala Indiana Gallorum, who died at Worms under Tiberius, has recently been accessed by the Musée départemental Dobrée in Nantes. A re-reading of this oldest known funerary monument of a Namnet provides the opportunity to place it among the stelae of foreign cavalrymen from Worms and Mainz, in the historical and military context of a Gaul in course of Romanization but still troubled by revolts such as, in 21, that of Andi and Turoni, then of the Trevir Iulius Florus and of the Eduan Iulius Sacrovir

PINK1-Interacting Proteins: Proteomic Analysis of Overexpressed PINK1

Recent publications suggest that the Parkinson's disease- (PD-) related PINK1/Parkin pathway promotes elimination of dysfunctional mitochondria by autophagy. We used tandem affinity purification (TAP), SDS-PAGE, and mass spectrometry as a first step towards identification of possible substrates for PINK1. The cellular abundance of selected identified interactors was investigated by Western blotting. Furthermore, one candidate gene was sequenced in 46 patients with atypical PD. In addition to two known binding partners (HSP90, CDC37), 12 proteins were identified using the TAP assay; four of which are mitochondrially localized (GRP75, HSP60, LRPPRC, and TUFM). Western blot analysis showed no differences in cellular abundance of these proteins comparing PINK1 mutant and control fibroblasts. When sequencing LRPPRC, four exonic synonymous changes and 20 polymorphisms in noncoding regions were detected. Our study provides a list of putative PINK1 binding partners, confirming previously described interactions, but also introducing novel mitochondrial proteins as potential components of the PINK1/Parkin mitophagy pathway

New insights into the complex role of mitochondria in Parkinson's disease

New discoveries providing insights into mitochondrial bioenergetics, their dynamic interactions as well as their role in cellular homeostasis have dramatically advanced our understanding of the neurodegenerative process of Parkinson's disease (PD). Respiratory chain impairment is a key feature in sporadic PD patients and there is growing evidence that links proteins encoded by PD-associated genes to disturbances in mitochondrial function. Against the backdrop of latest advances in the development of PD treatments that target mitochondria, we aim to give an overview of the literature published in the last three decades on the significance of mitochondria in the pathogenesis of PD. We describe the contribution of mitochondrial genome alterations and PD-associated genes to mitochondrial maintenance. We highlight mitophagy as a key mechanism in neurodegeneration. Moreover, we focus on the reciprocal interaction between alpha-synuclein aggregation and mitochondrial dysfunction. We discuss a novel trafficking pathway involving mitochondrial-derived vesicles within the context of PD and provide a synopsis of the most recently emerging topics in PD research with respect to mitochondria. This includes the relationship between mitochondria and cell-mediated immunity, the ER-mitochondria axis, sirtuin-mediated mitochondrial stress response and the role of micro RNAs in the aetiology of PD. In addition, recent studies have challenged the neuro-centric view of PD pathology, moving microglia and astrocytes into the research spotlight. Greater insights into these mechanisms may hold the key for the development of novel targeted therapies, addressing the need for a disease-modifying treatment, which has remained elusive to date

Tribological and mechanical properties of lubricant filled microcapsules in thermoplastic composites

Polymeric materials with long lifetime and low frictional energy loss are frequently required for a broad range of applications. Microencapsulation of lubricating oils by in-situ polymerization (melamine-formaldehyde) and interfacial polymerization (polyurethane/polyurea) was used to obtain free-flowing powders, which can be used as additive for thermoplastic materials resulting in microcapsule-containing self-lubricating composites. The specific functionality of such composites is achieved via portioned and localised release of the lubricant in the areas of the interface, which experiences the highest degrees of stress and wear due to the friction. Friction-triggered on-demand release of the lubricating oil results in materials with higher wear resistance and potentially leading to new products with prolonged lifetime. In this study, different ratios of microcapsules were added in polyoxymethylene (POM) and polybutylterephthalat (PBT) matrices by using laboratory scale twin-screw extruder resulting in self-lubricating composite materials. The effect of such modification on the tribological and mechanical properties of the thermoplastic composites were investigated. Rotational ball on disc tests were used to investigate the wear loss and coefficient of friction for the composites with varied microcapsule concentrations. Tensile tests revealed decreased mechanical stability for the composites with higher microcapsule content regardless of microcapsule wall material composition. Addition of 5&nbsp;wt.-&nbsp;% of encapsulated lubricant oil led to the substantial decrease of the frictional and wear coefficients. Further increase of encapsulated lubricant oil content to 10&nbsp;wt.-% had a major decreasing impact on the mechanical properties, whilst the effect on the tribological performance was rather small

ONG humanitaires et environnementales : l’alliance nécessaire ?

Christophe BuffetSi nous avons fait le choix de ce thème pour ce débat et plus largement pour ce nouveau dossier de la revue Humanitaire, c’est qu’on a le sentiment que depuis une bonne quarantaine d’années les ONG environnementales et humanitaires mènent leurs vies en parallèle. Les points de contact se font assez rares, les ONG de développement – plus promptes à prendre en compte les enjeux environnementaux – jouant d’une certaine manière le rôle d’acteurs pivots, capables de dialoguer avec..

Novel insights into Parkin-mediated mitochondrial dysfunction and neuroinflammation in Parkinson's disease

Mutations in PRKN cause the second most common genetic form of Parkinson's disease (PD)—a debilitating movement disorder that is on the rise due to population aging in the industrial world. PRKN codes for an E3 ubiquitin ligase that has been well established as a key regulator of mitophagy. Together with PTEN-induced kinase 1 (PINK1), Parkin controls the lysosomal degradation of depolarized mitochondria. But Parkin's functions go well beyond mitochondrial clearance: the versatile protein is involved in mitochondria-derived vesicle formation, cellular metabolism, calcium homeostasis, mitochondrial DNA maintenance, mitochondrial biogenesis, and apoptosis induction. Moreover, Parkin can act as a modulator of different inflammatory pathways. In the current review, we summarize the latest literature concerning the diverse roles of Parkin in maintaining a healthy mitochondrial pool. Moreover, we discuss how these recent discoveries may translate into personalized therapeutic approaches not only for PRKN-PD patients but also for a subset of idiopathic cases

Variants in Miro1 cause alterations of ER-mitochondria contact sites in fibroblasts from Parkinson's disease patients

Background: Although most cases of Parkinson´s disease (PD) are idiopathic with unknown cause, an increasing number of genes and genetic risk factors have been discovered that play a role in PD pathogenesis. Many of the PD‐associated proteins are involved in mitochondrial quality control, e.g., PINK1, Parkin, and LRRK2, which were recently identified as regulators of mitochondrial‐endoplasmic reticulum (ER) contact sites (MERCs) linking mitochondrial homeostasis to intracellular calcium handling. In this context, Miro1 is increasingly recognized to play a role in PD pathology. Recently, we identified the first PD patients carrying mutations in RHOT1, the gene coding for Miro1. Here, we describe two novel RHOT1 mutations identified in two PD patients and the characterization of the cellular phenotypes. Methods: Using whole exome sequencing we identified two PD patients carrying heterozygous mutations leading to the amino acid exchanges T351A and T610A in Miro1. We analyzed calcium homeostasis and MERCs in detail by live cell imaging and immunocytochemistry in patient‐derived fibroblasts. Results: We show that fibroblasts expressing mutant T351A or T610A Miro1 display impaired calcium homeostasis and a reduced amount of MERCs. All fibroblast lines from patients with pathogenic variants in Miro1, revealed alterations of the structure of MERCs. Conclusion: Our data suggest that Miro1 is important for the regulation of the structure and function of MERCs. Moreover, our study supports the role of MERCs in the pathogenesis of PD and further establishes variants in RHOT1 as rare genetic risk factors for neurodegeneration

Faithful SGCE imprinting in iPSC-derived cortical neurons: an endogenous cellular model of myoclonus-dystonia

In neuropathology research, induced pluripotent stem cell (iPSC)-derived neurons are considered a tool closely resembling the patient brain. Albeit in respect to epigenetics, this concept has been challenged. We generated iPSC-derived cortical neurons from myoclonus-dystonia patients with mutations (W100G and R102X) in the maternally imprinted ε-sarcoglycan (SGCE) gene and analysed properties such as imprinting, mRNA and protein expression. Comparison of the promoter during reprogramming and differentiation showed tissue-independent differential methylation. DNA sequencing with methylation-specific primers and cDNA analysis in patient neurons indicated selective expression of the mutated paternal SGCE allele. While fibroblasts only expressed the ubiquitous mRNA isoform, brain-specific SGCE mRNA and ε-sarcoglycan protein were detected in iPSC-derived control neurons. However, neuronal protein levels were reduced in both mutants. Our phenotypic characterization highlights the suitability of iPSC-derived cortical neurons with SGCE mutations for myoclonus-dystonia research and, in more general terms, prompts the use of iPSC-derived cellular models to study epigenetic mechanisms impacting on health and disease

Neurodegeneration and Neuroinflammation in Parkinson’s Disease: a Self-Sustained Loop

Purpose of Review: Neuroinflammation plays a significant role in Parkinson’s disease (PD) etiology along with mitochondrial dysfunction and impaired proteostasis. In this context, mechanisms related to immune response can act as modifiers at different steps of the neurodegenerative process and justify the growing interest in anti-inflammatory agents as potential disease-modifying treatments in PD. The discovery of inherited gene mutations in PD has allowed researchers to develop cellular and animal models to study the mechanisms of the underlying biology, but the original cause of neuroinflammation in PD is still debated to date. Recent Findings: Cell autonomous alterations in neuronal cells, including mitochondrial damage and protein aggregation, could play a role, but recent findings also highlighted the importance of intercellular communication at both local and systemic level. This has given rise to debate about the role of non-neuronal cells in PD and reignited intense research into the gut-brain axis and other non-neuronal interactions in the development of the disease. Whatever the original trigger of neuroinflammation in PD, what appears quite clear is that the aberrant activation of glial cells and other components of the immune system creates a vicious circle in which neurodegeneration and neuroinflammation nourish each other. Summary: In this review, we will provide an up-to-date summary of the main cellular alterations underlying neuroinflammation in PD, including those induced by environmental factors (e.g. the gut microbiome) and those related to the genetic background of affected patients. Starting from the lesson provided by familial forms of PD, we will discuss pathophysiological mechanisms linked to inflammation that could also play a role in idiopathic forms. Finally, we will comment on the potential clinical translatability of immunobiomarkers identified in PD patient cohorts and provide an update on current therapeutic strategies aimed at overcoming or preventing inflammation in PD. © 2022, The Author(s)