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Leishmania donovani lipophosphoglycan inhibits phagosomal maturation via action on membrane raft

PNPLA3 variant M148 causes resistance to starvation-mediated lipid droplet autophagy in human hepatocytes

The mechanism of how patatin-like phospholipase domain-containing protein 3 (PNPLA3) variant M148 is associated with increased risk of development of hepatic steatosis is still debated. Here, we propose a novel role of PNPLA3 as a key player during autophagosome formation in the process of lipophagy. A human hepatocyte cell line, HepG2 cells, expressing recombinant I148 or 148M, was used to study lipophagy under energy deprived conditions, and lipid droplet morphology was investigated using florescence microscopy, image analysis and biochemical assays. Autophagic flux was studied using the golden-standard of LC3-II turnover in combination with the well characterized GFP-RFP-LC3 vector. To discriminate between, perturbed autophagic initiation and lysosome functionality, lysosomes were characterized by Lysotracker staining and LAMP1 protein levels as well as activity and activation of cathepsin B. For validation, human liver biopsies genotyped for I148 and 148M were analyzed for the presence of LC3-II and PNPLA3 on lipid droplets. We show that the M148-PNPLA3 variant is associated with lipid droplets that are resistant to starvation-mediated degradation. M148 expressing hepatocytes reveal decreased autophagic flux and reduced lipophagy. Both I148-PNPLA3 and M148-PNPLA3 colocalize and interact with LC3-II, but the M148-PNPLA3 variant has lower ability to bind LC3-II. Together, our data indicate that PNPLA3 might play an essential role in lipophagy in hepatocytes and furthermore that the M148-PNPLA3 variant appears to display a loss in this activity, leading to decreased lipophagy

The European Union and the autonomy of sub-national authorities: Towards an analysis of constraints and opportunities in sub-national decision-making

This paper explores what is known about the actual impact of the EU on sub-national (local and regional) government. Existing research on the impact of the EU on sub-national authorities appears to have a strong emphasis on the positive effects or the opportunities that emanate from the EU. By contrast, studies on European constraints that limit sub-national autonomy are rather scarce. Moreover, many studies fail to take sub-national government itself as the object of analysis, as a result of which most conclusions are rather hypothetical in nature. Trying to fill the lacunas in the existing literature, the authors present a conceptual framework that includes three dimensions along which the EU might influence sub-national decision-making: by enforcement or invitation, by hampering or improvement, and by obstruction or enabling. A pilot study found that nearly each of these types of influence was present, indicating the conceptual sensitivity and empirical relevance of the three dimensions. © 2006 Taylor & Francis