45 research outputs found

    Phagocytosis and digestion of pH-sensitive fluorescent dye (Eos-FP) transfected E. coli in whole blood assays from patients with severe sepsis and septic shock

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    The function of phagocytic and antigen presenting cells is of crucial importance to sustain immune competence against infectious agents as well as malignancies. We here describe a reproducible procedure for the quantification of phagocytosis by leukocytes in whole blood. For this, a pH-sensitive green-fluorescent protein- (GFP) like dye (Eos-FP) is transfected into infectious microroganisms. After UV-irradiation, the transfected bacteria emit green (≈5160 nm) and red (≈581 nm) fluorescent light at 490 nm excitation. Since the red fluorescent light is sensitive to acidic pH, the phagocytosed bacteria stop emitting red fluorescent light as soon as the phagosomes fuse with lysosomes. The green fluorescence is maintained in the phagolysosome until pathogen degradation is completed. Fluorescence emission can be followed by flow cytometry with filter settings documenting fluorescence 1 (FL 1, FITC) and fluorescence 2 (FL 2, phycoerythrin, PE). Eos-FP transfected bacteria can also be traced within phagocytes using microscopical techniques. A standardized assay has been developed which is suitable for clinical studies by providing clinicians with syringes pre-filled with fixed and appropriately UV-irradiated Eos-FP E. coli (TruCulture™). After adding blood or body fluids to these containers and starting the incubation at 37°C, phagocytosis by granulocytes proceeds over time. Cultures can be terminated at a given time by lysing red blood cells followed by flow cytometry. A pilot study demonstrated that Eos-FP E. coli phagocytosis and digestion was up-regulated in the majority of patients with either severe sepsis or septic shock as compared to healthy donors (p < 0.0001 after o/n incubation). Following treatment with recombinant human granulocyte colony-stimulating factor (rhG-CSF) in selected patients with sepsis, phagolysosome fusion appeared to be accelerated

    Measurement of Angular Coefficients of BˉDνˉ\bar{B} \to D^* \ell \bar{\nu}_\ell: Implications for Vcb|V_{cb}| and Tests of Lepton Flavor Universality

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    We measure the complete set of angular coefficients JiJ_i for exclusive BˉDνˉ\bar{B} \to D^* \ell \bar{\nu}_\ell decays (=e,μ\ell = e, \mu). Our analysis uses the full 711fb1711\,\mathrm{fb}^{-1} Belle data set with hadronic tag-side reconstruction. The results allow us to extract the form factors describing the BDB \to D^* transition and the Cabibbo-Kobayashi-Maskawa matrix element Vcb|V_{\rm cb}|. Using recent lattice QCD calculations for the hadronic form factors, we find Vcb=(41.0±0.7)×103|V_{\rm cb}| = (41.0 \pm 0.7) \times 10^3 using the BGL parameterization, compatible with determinations from inclusive semileptonic decays. We search for lepton flavor universality violation as a function of the hadronic recoil parameter ww, and investigate the differences of the electron and muon angular distributions. We find no deviation from Standard Model expectations

    Positive Myonen in _#alpha#-Eisen bei tiefen Temperaturen

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    SIGLETIB Hannover: DR 4988 / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekDEGerman

    Themes and Research Directions in Privacy-Sensitive Robotics

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    Privacy is crucial for healthy relationships, but robots will impact our privacy in new ways - this warrants a new area of research. This paper presents work from the first workshop on privacy-sensitive robotics. We identify the seven research themes that should comprise privacy-sensitive robotics research in the near future: data privacy; manipulation and deception; trust; blame and transparency; legal issues; domains with special privacy concerns; and privacy theory. We intend for the research directions proposed for each of these themes to serve as a roadmap for privacy-sensitive robotics research. © 2018 IEEE
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