Mass spectrometry-based analytical tools for the molecular protein characterization of human plasma lipoproteins

Lipoproteins are a heterogeneous population of blood plasma particles composed of apolipoproteins and lipids. Lipoproteins transport exogenous and endogenous triglycerides and cholesterol from sites of absorption and formation to sites of storage and usage. Three major classes of lipoproteins are distinguished according to their density: high-density (HDL), low-density (LDL) and very low-density lipoproteins (VLDL). While HDLs contain mainly apolipoproteins of lower molecular weight, the two other classes contain apolipoprotein B and apolipoprotein (a) together with triglycerides and cholesterol. HDL concentrations were found to be inversely related to coronary heart disease and LDL/VLDL concentrations directly related. Although many studies have been published in this area, few have concentrated on the exact protein composition of lipoprotein particles. Lipoproteins were separated by density gradient ultracentrifugation into different subclasses. Native gel electrophoresis revealed different gel migration behaviour of the particles, with less dense particles having higher apparent hydrodynamic radii than denser particles. Apolipoprotein composition profiles were measured by matrix-assisted laser desorption/ionization-mass spectrometry on a macromizer instrument, equipped with the recently introduced cryodetector technology, and revealed differences in apolipoprotein composition between HDL subclasses. By combining these profiles with protein identifications from native and denaturing polyacrylamide gels by liquid chromatography-tandem mass spectrometry, we characterized comprehensively the exact protein composition of different lipoprotein particles. We concluded that the differential display of protein weight information acquired by macromizer mass spectrometry is an excellent tool for revealing structural variations of different lipoprotein particles, and hence the foundation is laid for the screening of cardiovascular disease risk factors associated with lipoproteins.

Low-density lipoprotein particle diameter and mortality: the Ludwigshafen Risk and Cardiovascular Health Study.

AIMS The aim of the study was to examine whether differences in average diameter of low-density lipoprotein (LDL) particles were associated with total and cardiovascular mortality. METHODS AND RESULTS We studied 1643 subjects referred to coronary angiography, who did not receive lipid-lowering drugs. During a median follow-up of 9.9 years, 398 patients died, of these 246 from cardiovascular causes. We calculated average particle diameters of LDL from the composition of LDL obtained by β-quantification. When LDL with intermediate average diameters (16.5-16.8 nm) were used as reference category, the hazard ratios (HRs) adjusted for cardiovascular risk factors for death from any cause were 1.71 (95% CI: 1.31-2.25) and 1.24 (95% CI: 0.95-1.63) in patients with large (>16.8 nm) or small LDL (<16.5 nm), respectively. Adjusted HRs for death from cardiovascular causes were 1.89 (95% CI: 1.32-2.70) and 1.54 (95% CI: 1.06-2.12) in patients with large or small LDL, respectively. Patients with large LDL had higher concentrations of the inflammatory markers interleukin (IL)-6 and C-reactive protein than patients with small or intermediate LDL. Equilibrium density gradient ultracentrifugation revealed characteristic and distinct profiles of LDL particles in persons with large (approximately even distribution of intermediate-density lipoproteins and LDL-1 through LDL-6) intermediate (peak concentration at LDL-4) or small (peak concentration at LDL-6) average LDL particle diameters. CONCLUSIONS Calculated LDL particle diameters identify patients with different profiles of LDL subfractions. Both large and small LDL diameters are independently associated with increased risk mortality of all causes and, more so, due to cardiovascular causes compared with LDL of intermediate size.

Federalism and the Concept of Political Territoriality: Towards an Analytical Framework for Comparative Territorial Politics

The core issues comparative territorial politics addresses are how and why territory is used to delimit, maintain, or create political power; and with what kind of consequences for efficiency (output) and legitimacy (input). The aim of this article is to integrate various research strands into the comparative study of territorial politics, with federal studies at its core. As an example of a conceptual payoff, ‘political territoriality’ refers the observer to three dimensions of the strategic use of areal boundaries for political power. By focusing on territory as a key variable of political systems, the actors, processes and institutions are first analytically separated and continuously measured, enhancing internal validity, and then theoretically integrated, which allows more valid external inferences than classic, legal-institutionalist federal studies. After discussing the boundaries and substance of comparative territorial politics as a federal discipline, political territoriality is developed towards an analytical framework applicable to politics at any governmental level. The claims are modest: political territoriality does not serve so much as an explanatory concept as rather an ‘attention-directing device’ for federal studies.

Evénements hydrologiques extrêmes

Ce matériel didactique comprend des contenus hydrologiques importants pour l'enseignement de la géographie au niveau secondaire II. Les fiches thématiques, utilisables de manière flexible, ainsi que l'e-book complémentaire, permettent d'aborder les problématiques hydrologiques actuelles de manière approfondie et analytique.

"Montrer au linguiste ce qu'il fait". Une analyse épistémologique du structuralisme européen (Hjelmslev, Jakobson, Martinet, Benveniste) dans sa filiation saussurienne

Cette thèse consiste en une analyse épistémologique comparée et très détaillée de l’ensemble du corpus saussurien publié ainsi que d’une portion très significative des oeuvres de Hjelmslev, Jakobson, Martinet et Benveniste. Il s’agit de montrer qu’en dépit d’une filiation revendiquée le structuralisme européen n’est pas saussurien, et par là de faire apparaître, par contrecoup, la spécificité de la problématique saussurienne, ainsi que ses enjeux pour la linguistique et plus largement pour les sciences de l’humain. La problématique saussurienne avait permis, pour la première fois dans l’histoire de la linguistique, une appréhension théorique de la langue. La problématique structuraliste est en revanche entièrement empirique, de sorte que sa scientificité relève en réalité d’une idéologie scientifique, au sens de Georges Canguilhem. Le point nodal de cette radicale différence de problématique est l’absence de théorisation structuraliste du rapport son/sens, et corrélativement la mécompréhension du concept saussurien de système. Celui-ci devient alors structure, c’est-à-dire, comme nous tentons de le faire apparaître, appréhension structurale d’un objet dont la définition commune et évidente (celle de la langue comme instrument de communication) n’est pas remise en cause. A la problématique étiologique saussurienne, constitutive du concept de langue, répond ainsi une problématique analytique qui conduit quant à elle à la construction d’un objet (forme ou structure) en lieu et place d’un concept. Plus précisément, la problématique structuraliste est idiomologique. Elle manque ainsi la distinction entre langue et idiome dont nous tentons dès lors de démontrer la nécessité et le caractère constitutif de la théorisation de la langue et, au-delà, du langage, notamment dans le cadre d’une articulation entre linguistique et psychanalyse.

Transitionen im Jugend- und jungen Erwachsenenalter: Ergebnisse der Schweizer Längsschnittstudie TREE

Nach dem ersten, 2011 erschienenen Band versammelt der vorliegende zweite wiederum eine Auswahl von Beiträgen, die auf Analysen der Daten der Schweizer Längsschnittstudie TREE (Transitionen von der Erstausbildung ins Erwerbsleben) basieren. Die kritischen Übergänge im Jugend- und jungen Erwachsenenalter werden aus soziologischen, ökonomischen, psychologischen und erziehungswissenschaftlichen Blickwinkeln beleuchtet. Die Beiträge widerspiegeln damit eindrücklich das analytische und interdisziplinäre Potenzial der TREE-Daten. Thematisch steht der langfristige Einfluss der sozialen Herkunft auf Bildungs- und Erwerbsverläufe, insbesondere auf den Zugang zu höherer Bildung, im Zentrum.

The Gas6-Axl Interaction Mediates Endothelial Uptake of Platelet Microparticles.

Upon activation, platelets release plasma-membrane derived microparticles (PMPs) exposing phosphatidylserine (PS) on their surface. The function and clearance mechanism of these MPs are incompletely understood. As they are pro-coagulant and potentially pro-inflammatory, rapid clearance from the circulation is essential for prevention of thrombotic diseases. The tyrosine kinase receptors Tyro3, Axl and Mer (TAMs) and their ligands protein S and Gas6 are involved in the uptake of PS-exposing apoptotic cells in macrophages and dendritic cells. Both TAMs and their ligands are expressed in the vasculature, the functional significance of which is poorly understood. In this study we investigated how vascular TAMs and their ligands may mediate endothelial uptake of PMPs. PMPs, generated from purified human platelets, were isolated by ultracentrifugation and labeled with biotin or PKH67. The uptake of labeled MPs in the presence of protein S and Gas6 in human aortic endothelial cells (HAEC) and human umbilical vein endothelial cells (HUVEC) was monitored by flow cytometry, western blotting and confocal/electron microscopy. We found that both endothelial cell types can phagocytose PMPs, and using TAM-blocking antibodies or siRNA knock-down of individual TAMs we show that the uptake is mediated by endothelial Axl and Gas6. As circulating PMPs-levels were not altered in Gas6-/- mice compared to Gas6+/+ mice, we hypothesize that the Gas6-mediated uptake is not a means to clear the bulk of circulating PMPs but may serve to phagocytose PMPs locally generated at sites of platelet activation and as a way to affect endothelial responses.