Monitoring the Formation of Autophagosomal Precursor Structures in Yeast Saccharomyces cerevisiae

The budding yeast Saccharomyces cerevisiae is a powerful and versatile model organism for studying multiple aspects of the biology of eukaryotic cells, including the molecular principles underlying autophagy. One of the unique advantages of this unicellular system is its amenability to genetic and biochemical approaches, which had a pivotal role in the discovery and characterization of most of the autophagy-related (Atg) proteins, the central players of autophagy. The relevance of investigating autophagy in this cell model lies in the high conservation of this pathway among eukaryotes, i.e., most of the yeast Atg proteins possess one or more mammalian orthologs. In addition to the experimental advantages, a very large collection of reagents keeps S. cerevisiae in a leading position for the study of the molecular mechanism and regulation of autophagy. In this chapter, we describe fluorescence microscopy and biochemical methods that allow to monitor in vivo the assembly the of Atg machinery, a key step of autophagy. These approaches can be very useful to those researchers that would like to assess the progression of the autophagosomal precursor structure formation under various conditions, in the presence of specific Atg protein mutants or in the absence of other factors

Personalized Content Preparation and Delivery for Universal Multimedia Access

This paper introduces a personalized content preparation and delivery framework for universal multimedia access. This framework has a wealth of potential applications in broadcasting, the World Wide Web and mobile telecommunication. Content preparation deals with the generation of summaries by automatic video analysis techniques, and with the annotation of content using MPEG7 for search and retrieval and SMIL for interactive video. The use of open standards favors interoperability with third-party applications and upcoming hardware. Content delivery handles individual user preferences, content selection, real-time content adaptation and streaming in a seamless way. The proposed framework has been implemented in the ECfunded R&D project PERSEO and tested by selected groups of end-users. Personalization, video summaries and the overall usability have been positively evaluated