Real time imaging of calcium-induced localized proteolytic activity after axotomy and its relation to growth cone formation

The emergence of a neuronal growth cone from a transected axon is a necessary step in the sequence of events that leads to successful regeneration. Yet, the molecular mechanisms underlying its formation after axotomy are unknown. In this study, we show by real time imaging of the free intracellular Ca2+ concentration, of proteolytic activity, and of growth cone formation that the activation of localized and transient Ca2+-dependent proteolysis is a necessary step in the cascade of events that leads to growth cone formation. Inhibition of this proteolytic activity by calpeptin, a calpain inhibitor, abolishes growth cone formation. We suggest that calpain plays a central role in the reorganization of the axon's cytoskeleton during its transition from a stable differentiated structure into a dynamically extending growth cone.     

A suite of Gateway cloning vectors for high-throughput genetic analysis in Saccharomyces cerevisiae

In the post-genomic era, academic and biotechnological research is increasingly shifting its attention from single proteins to the analysis of complex protein networks. This change in experimental design requires the use of simple and experimentally tractable organisms, such as the unicellular eukaryote Saccharomyces cerevisiae, and a range of new high-throughput techniques. The Gateway system has emerged as a powerful high-throughput cloning method that allows for the in vitro recombination of DNA with high speed, accuracy and reliability. Two Gateway-based libraries of overexpression plasmids containing the entire complement of yeast open reading frames (ORFs) have recently been completed. In order to make use of these powerful resources, we adapted the widely used pRS series of yeast shuttle vectors for use in Gateway-based cloning. The resulting suite of 288 yeast Gateway vectors is based upon the two commonly used GPD and GAL1 promoter expression systems that enable expression of ORFs, either constitutively or under galactose-inducible conditions. In addition, proteins of interest can be fused to a choice of frequently used N- or C-terminal tags, such as EGFP, ECFP, EYFP, Cerulean, monomeric DsRed, HA or TAP. We have made this yeast Gateway vector kit available to the research community via the non-profit Addgene Plasmid Repository (http://www.addgene.org/yeast_gateway).     

Energy consumption model in multicore architectures with variable frequency

Models extending Amdahl’s law have been developed to study the behavior of parallel programs energy consumption. In addition, it has been shown that energy consumption of those programs also relies on the layout of the resources on the chip, such as power supply. Other extensions over Amdahl’s law have been conducted to study the behavior of parallel programs speedup for frequency variable processors. Previous models have focused on the use of Turbo Boost in the parallel regions of a program, without considering that Turbo Boost also affects the sequential regions. Hence, we present a model to analyze energy consumption of parallel programs executed on Intel multicore processors with Turbo Boost frequencies to cover this gap. The model is an extension to Amdahl’s law, and it is validated with a double-precision matrix multiplication running on Intel multicore processors that enable Turbo Boost technology.