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    Real-time image processing for label-free enrichment of Actinobacteria cultivated in picolitre droplets
    (London [u.a.] : Royal Society of Chemistry, 2013) Zang, E.; Brandes, S.; Tovar, M.; Martin, K.; Mech, F.; Horbert, P.; Henkel, T.; Figge, M.T.; Roth, M.
    The majority of today's antimicrobial therapeutics is derived from secondary metabolites produced by Actinobacteria. While it is generally assumed that less than 1% of Actinobacteria species from soil habitats have been cultivated so far, classic screening approaches fail to supply new substances, often due to limited throughput and frequent rediscovery of already known strains. To overcome these restrictions, we implement high-throughput cultivation of soil-derived Actinobacteria in microfluidic pL-droplets by generating more than 600000 pure cultures per hour from a spore suspension that can subsequently be incubated for days to weeks. Moreover, we introduce triggered imaging with real-time image-based droplet classification as a novel universal method for pL-droplet sorting. Growth-dependent droplet sorting at frequencies above 100 Hz is performed for label-free enrichment and extraction of microcultures. The combination of both cultivation of Actinobacteria in pL-droplets and real-time detection of growing Actinobacteria has great potential in screening for yet unknown species as well as their undiscovered natural products.
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    Charlemagne's summit canal: An early medieval hydro-engineering project for passing the Central European Watershed
    (San Francisco, CA : Public Library of Science, 2014) Zielhofer, C.; Leitholdt, E.; Werther, L.; Stele, A.; Bussmann, J.; Linzen, S.; Schneider, M.; Meyer, C.; Berg-Hobohm, S.; Ettel, P.
    The Central European Watershed divides the Rhine-Main catchment and the Danube catchment. In the Early Medieval period, when ships were important means of transportation, Charlemagne decided to link both catchments by the construction of a canal connecting the Schwabian Rezat and the Altmü hl rivers. The artificial waterway would provide a continuous inland navigation route from the North Sea to the Black Sea. The shortcut is known as Fossa Carolina and represents one of the most important Early Medieval engineering achievements in Europe. Despite the important geostrategic relevance of the construction it is not clarified whether the canal was actually used as a navigation waterway. We present new geophysical data and in situ findings from the trench fills that prove for the first time a total length of the constructed Carolingian canal of at least 2300 metres. We have evidence for a conceptual width of the artificial water course between 5 and 6 metres and a water depth of at least 60 to 80 cm. This allows a crossing way passage of Carolingian cargo scows with a payload of several tons. There is strong evidence for clayey to silty layers in the trench fills which reveal suspension load limited stillwater deposition and, therefore, the evidence of former Carolingian and post-Carolingian ponds. These findings are strongly supported by numerous sapropel layers within the trench fills. Our results presented in this study indicate an extraordinarily advanced construction level of the known course of the canal. Here, the excavated levels of Carolingian trench bottoms were generally sufficient for the efficient construction of stepped ponds and prove a final concept for a summit canal. We have evidence for the artificial Carolingian dislocation of the watershed and assume a sophisticated Early Medieval hydrological engineering concept for supplying the summit of the canal with adequate water.
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    Vinculin binding angle in podosomes revealed by high resolution microscopy
    (San Francisco, CA : Public Library of Science, 2014) Walde, M.; Monypenny, J.; Heintzmann, R.; Jones, G.E.; Cox, S.
    Podosomes are highly dynamic actin-rich adhesive structures formed predominantly by cells of the monocytic lineage, which degrade the extracellular matrix. They consist of a core of F-actin and actin-regulating proteins, surrounded by a ring of adhesion-associated proteins such as vinculin. We have characterised the structure of podosomes in macrophages, particularly the structure of the ring, using three super-resolution fluorescence microscopy techniques: stimulated emission depletion microscopy, structured illumination microscopy and localisation microscopy. Rather than being round, as previously assumed, we found the vinculin ring to be created from relatively straight strands of vinculin, resulting in a distinctly polygonal shape. The strands bind preferentially at angles between 116° and 135°. Furthermore, adjacent vinculin strands are observed nucleating at the corners of the podosomes, suggesting a mechanism for podosome growth.