Schematic description of the experimental design used to measure cAMP nanocompartments on a single molecule resolution.

<p>The PDE molecule is modeled as an absorbing sphere of radius <i>R</i>_PDE and the sensor protein attached adjacent (distance <i>d</i>) is used to measure the cAMP concentration.</p

Fusion of PDE4A1 to Epac1-camps generates cAMP nanodomains in living cells.

<p>(a) and (c) Representative traces of the normalized FRET (YFP/CFP) ratio of the indicated constructs. Increases of cAMP were obtained by activation of endogenous <i>β</i>-receptors by isoproterenol (a) or upon stimulation with the direct activators of adenylyl cyclase forskolin (c) and (d) Amplitude of the cAMP response elicited by isoproterenol (b) or forskolin (d) expressed as a percentage of maximal stimulation induced by rolipram. Data are shown as means ± s.e.m. of at least 6 independent experiments. Differences vs. Epac1-camps were statistically significant by one-way ANOVA followed by Bonferroni post hoc-test **, P < 0, 001 ****, P < 0,00001.</p

Binding curves of the free sensor protein Epac1-camps + PDE (blue) and the fusion protein Epac1-camps-PDE4A1 (red).

<p>The overall PDE activity is equal in both experiments, indicating that the right shift of the binding curve is solely caused by local PDE degradation. The curves were determined by fitting <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0174856#pone.0174856.e028" target="_blank">Eq (18)</a> to the corresponding experimental data. The distance between sensor and absorbing enzyme was set to <i>d</i> = 0nm as for an ideal sensor-enzyme construct. The fit of the free sensor data (blue) yields the dissociation constant of the sensor <i>K</i>_<i>D</i> = 7.3 ± 0.66<i>μ</i>M (goodness of fit: <i>χ</i>² = 3.2, <i>p</i> = 0.66). From the sensor-enzyme construct (red) we obtain the absorptive action of a PDE <i>η</i>₂ = 6.1 ± 1.4, (<i>χ</i>² = 9.6, <i>p</i> = 0.08). The flooding of the nanocompartment as predicted by our model could not be observed in the experiments.</p

Additional file 1: Table S1. of Renal function in patients with non-dialysis chronic kidney disease receiving intravenous ferric carboxymaltose: an analysis of the randomized FIND-CKD trial

Baseline characteristics (ITT population). (DOCX 12 kb

Additional file 2: Table S2. of Renal function in patients with non-dialysis chronic kidney disease receiving intravenous ferric carboxymaltose: an analysis of the randomized FIND-CKD trial

Mean (SD) change in estimated GFR (eGFR) from baseline to month 12 for subpopulations of patients with eGFR values both time points. (DOCX 12 kb

Additional file 1: of Copeptin levels and commonly used laboratory parameters in hospitalised patients with severe hypernatraemia - the “Co-MED study”

Clinical diagnostic algorithm for the differential diagnosis of hypernatremia. (TIFF 143 kb