Nachrichten aus der Brüder-Gemeine, 1828, no. 01

The Moravian Church's monthly journal of its worldwide activities, including in Labrador, published from 1819-94

Bondholder Coercion: The Problem of Constrained Choice in Debt Tender Offers and Recapitalizations

<p>The effect MATE1, MATE2 and PMAT inhibitors on the [³H]pentamidine accumulation in hCMEC/D3 (A) and bEnd.3 (B) cells. Cells were incubated with MATE1 inhibitor famotidine (1 μM), MATE2 inhibitor nifekalant (3 μM) or PMAT inhibitor lopinavir (2 μM) and no significant differences were observed compared to control. All data expressed as mean ± SEM, n =.4 passages of cells, with 6 replicates (wells) per timepoint per plate. Data were analysed using two-way ANOVA with SigmaPlot 13.0.</p

Expression of OCT1 was detected by Transmission Electron Microscopy and immunogold labelling.

<p>hCMEC/D3 (p28) and bEnd.3 cells (p23) were grown to confluency on transwell polycarbonate membrane and stained for OCT1 using the method described. Gold nanoparticles on the membranes (arrows) were counted in the 60 sequential images acquired at 11500 x magnification. Significantly increased expression of OCT1 was found on the luminal membrane compared to the abluminal membrane. ***p<0.001 using student’s t-test.</p

Organic cation transporter 1 (OCT1) is involved in pentamidine transport at the human and mouse blood-brain barrier (BBB) - Fig 10

<p>Confocal microscopy image of MRP4 expression in confluent hCMEC/D3 cells (A) and bEnd.3 cells (B). No fluorescent signals were detected when using MRP4 antibody [M4I-10] from Abcam at 1:200 dilution and goat anti-rat Alexa Fluor^® 488 secondary antibody (Abcam, ab181448) at 1:200 dilution.</p

BCRP expression in confluent hCMEC/D3 cells and bEnd.3 cells.

<p>Confocal microscopy image of BCRP expression in confluent hCMEC/D3 cells (A) and bEnd.3 cells (B) was not detected in the plasma membranes when using anti-BCRP antibody (New England Biolabs, 4477S diluted at 1:200)—Western blot band of BCRP was detected at 143 kD which corresponds to the dimer version of BCRP in positive control (C). No band was observed at 70 kD (monomer version of BCRP). Tubulin (55 kD) was used as a loading control. Lane 1 –hCMEC/D3 passage 28, Lane 2- hCMEC/D3 passage 33, Lane 3- bEnd.3 passage 18, Lane 4- bEnd.3 passage 23, Lane 5 –brain endothelial cells isolated from mouse (positive control).</p

The transporter inhibitors used in this study along with [³H]pentamidine.

<p>All inhibitors were used in the presence of 0.05% DMSO and used at the published concentration ranges where they affect transporter activity.</p

Systematic Investigation of the Physicochemical Factors That Contribute to the Toxicity of ZnO Nanoparticles

ZnO nanoparticles (NPs) are prone to dissolution, and uncertainty remains whether biological/cellular responses to ZnO NPs are solely due to the release of Zn²⁺ or whether the NPs themselves have additional toxic effects. We address this by establishing ZnO NP solubility in dispersion media (Dulbecco’s modified Eagle’s medium, DMEM) held under conditions identical to those employed for cell culture (37 °C, 5% CO₂, and pH 7.68) and by systematic comparison of cell–NP interaction for three different ZnO NP preparations. For NPs at concentrations up to 5.5 μg ZnO/mL, dissolution is complete (with the majority of the soluble zinc complexed to dissolved ligands in the medium), taking ca. 1 h for uncoated and ca. 6 h for polymer coated ones. Above 5.5 μg/mL, the results are consistent with the formation of zinc carbonate, keeping the solubilized zinc fixed to 67 μM of which only 0.45 μM is as free Zn²⁺, i.e., not complexed to dissolved ligands. At these relatively high concentrations, NPs with an aliphatic polyether-coating show slower dissolution (i.e., slower free Zn²⁺ release) and reprecipitation kinetics compared to those of uncoated NPs, requiring more than 48 h to reach thermodynamic equilibrium. Cytotoxicity (MTT) and DNA damage (Comet) assay dose–response curves for three epithelial cell lines suggest that dissolution and reprecipitation dominate for uncoated ZnO NPs. Transmission electron microscopy combined with the monitoring of intracellular Zn²⁺ concentrations and ZnO–NP interactions with model lipid membranes indicate that an aliphatic polyether coat on ZnO NPs increases cellular uptake, enhancing toxicity by enabling intracellular dissolution and release of Zn²⁺. Similarly, we demonstrate that needle-like NP morphologies enhance toxicity by apparently frustrating cellular uptake. To limit toxicity, ZnO NPs with nonacicular morphologies and coatings that only weakly interact with cellular membranes are recommended