A novel reverse transduction adenoviral array for the functional analysis of shRNA libraries

<p>Abstract</p> <p>Background</p> <p>The identification of novel drug targets by assessing gene functions is most conveniently achieved by high-throughput loss-of-function RNA interference screening. There is a growing need to employ primary cells in such screenings, since they reflect the physiological situation more closely than transformed cell lines do. Highly miniaturized and parallelized approaches as exemplified by reverse transfection or transduction arrays meet these requirements, hence we verified the applicability of an adenoviral microarray for the elucidation of gene functions in primary cells.</p> <p>Results</p> <p>Here, we present microarrays of infectious adenoviruses encoding short hairpin RNA (shRNA) as a new tool for gene function analysis. As an example to demonstrate its application, we chose shRNAs directed against seven selected human protein kinases, and we have performed quantitative analysis of phenotypical responses in primary human umbilical vein cells (HUVEC). These microarrays enabled us to infect the target cells in a parallelized and miniaturized procedure without significant cross-contamination: Viruses were reversibly immobilized in spots in such a way that the seeded cells were confined to the area of the viral spots, thus simplifying the subsequent addressing of genetically modified cells for analysis. Computer-assisted image analysis of fluorescence images was applied to analyze the cellular response after shRNA expression. Both the expression level of knock-down target proteins as well as the functional output as measured by caspase 3 activity and DNA fractionation (TUNEL) were quantified.</p> <p>Conclusion</p> <p>We have developed an adenoviral microarray technique suitable for miniaturized and parallelized analysis of gene function. The practicability of this technique was demonstrated by the analysis of several kinases involved in the activation of programmed cell death, both in tumor cells and in primary cells.</p

Anarchy in the UJ: Coordination mechanisms for minimizing the number of late jobs

We consider the distributed scheduling problem on parallel machines with the central objective of maximizing the number of on-time jobs. Jobs are self-interested utility-maximizers that can choose the machines they are processed on and are exclusively interested in reducing their own private objective function. Each machine processes the jobs according to a local policy. We discuss Nash equilibria in the resulting schedules and perform a thorough analysis of the resulting (absolute) prices of anarchy for various parallel machine environments, utilities of the agents, and local policies of the machines. We show that local policies that are based on simple sorting-based procedures like shortest processing time first (SPT) and earliest due date first (EDD) lead to big losses in welfare compared to the global optimum. However, when employing Moore-Hodgson's algorithm as a local policy, we can prove a price of anarchy of (2m−1)/m for identical machines and a price of anarchy of 2 for related and unrelated parallel machines. Moreover, we show how these results can be used to prove approximation ratios for greedy scheduling algorithms. This paper is the first to prove approximation ratios for two greedy scheduling procedures, turning them from simple heuristics into actual approximation ratios with a provable approximation ratio

Deutsche S3-Leitlinie Behandlung von Angststörungen

Die deutsche S3-Leitline zur Behandlung von Angststörungen (Panikstörung/Agoraphobie, generalisierte Angststörung, soziale Phobie, spezifische Phobie) bei Erwachsenen wurde unter Beratung und Moderation durch die Arbeitsgemeinschaft der Wissenschaftlichen Medizinischen Fachgesellschaften (AWMF) von einem Gremium erstellt, das 20 Fachverbände und andere Organisationen aus den Bereichen Psychotherapie, Psychologie, psychosomatische Medizin, Psychiatrie und Allgemeinmedizin sowie Patientenvertreter und Selbsthilfeorganisationen umfasst. Die Empfehlungen dieser Leitlinie basieren auf einer Sichtung der Evidenz der verfügbaren randomisierten klinischen Studien zu Angststörungen nach ICD/DSM und einer Synthese der Empfehlungen anderer Leitlinien