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The US-tubes then were debundled by Birch reduction using sodium metal in tetrahydrofuran [1]. Briefly, 100?mg of US-tubes were added to 100?ml of aqueous 5.0?mm GdCl3 or DyCl3 solution prepared with HPLC-grade water (1?mg of US-tube nanocapsules R428 ml?1 solution, pH 2.5) and bath sonicated for 1?h. The lanthanide salts (99.99% pure) were anhydrous powders (Aldrich, Saint Louis, MO, USA). The solution was allowed to stand overnight. The GNTs and DNTs flocculated to the bottom of the reaction vessel and were washed with HPLC-grade water (pH 7.0) on a coarse fritted-glass filter until the filtrate showed no detectible levels (E-64 www.selleckchem.com/products/jq1.html signaling from, for example, a neuronal injury. In vitro, this activation is induced by lipopolysacchride (LPS) endotoxin, a substance formed from the cell wall of Gram-negative bacteria and well known for the activation of microglia cells by mimicking a septic shock [39-41]. While these cells are prime candidates as early biomarkers of neurodegeneration, they also serve well as a model cellular system for the assessment of new contrast agents. For this study, Bv2 cells were grown in 75?cm2 tissue culture flasks with Dulbecco's modified Eagle's medium (Invitrogen Corp., Carlsbad, CA, USA). The medium was supplemented with 10% nonessential amino acids, 10% heat inactivated fetal bovine serum (Atlanta Biological, Alanta, GA, USA), 1% antibiotics/antimyotics (Gibco, Invitrogen Corp., Carlsbad, CA, USA) and 0.1% gentamycin (Invitrogen Corp., Carlsbad, CA, USA). Cells were kept at a temperature of 37?��C and under 5% CO2. Six-well plates (10?cm2) were seeded with approximately 40 000 cells per well. Cells were given 24?h to attach and grow before exposure to treatment. Based on the Ln3+ concentration in the stock solution, as seen in Table?1, cell media and US-tube solution were adjusted so that cells were exposed to 17?mm Ln3+ concentrations. This concentration was chosen based on studies by Tang et al. [9] for which viability of the evaluated macrophage cell line (J774A.