A multi-phenotypic imaging screen to identify bacterial effectors by exogenous expression in a HeLa cell line

We present a high-content screen (HCS) for the simultaneous analysis of multiple phenotypes in HeLa cells expressing an autophagy reporter (mcherry-LC3) and one of 209 GFP-fused proteins from the Crohn’s Disease (CD)-associated bacterium, Adherent Invasive E. coli (AIEC) strain LF82. Using automated confocal microscopy and image analysis (CellProfiler), we localised GFP fusions within cells, and monitored their effects upon autophagy (an important innate cellular defence mechanism), cellular and nuclear morphology, and the actin cytoskeleton. This data will provide an atlas for the localisation of 209 AIEC proteins within human cells, as well as a dataset to analyse their effects upon many aspects of host cell morphology. We also describe an open-source, automated, image-analysis workflow to identify bacterial effectors and their roles via the perturbations induced in reporter cell lines when candidate effectors are exogenously expressed

Water with Excess Electric Charge

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Water dropping from a biased metal needle acquires net electric charge that can be stored within. a Faraday cup. Positive (negative) water is obtained from a positive (negative) needle, and its charge largely exceeds the Rayleigh limit. Water drops transform into water threads at V < -9.5 kV, even under field strengths much lower than those used in electrospray, electrowetting, or electrospinning experiments. Surface tension of electrified water decreases as the positive or negative charge increases, but its density and viscosity are the same as those of neutral water. Water drops falling in the air through a biased noncontacting metal ring also acquire charge but with opposite sign to the metal potential, in agreement with a recent model for the electrification of insulators or isolated metals, based on charge transfer coupled to water evaporation, adsorption, and desorption.115221122611232Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP