Alzheimer's disease (AD) is an age-related neurodegenerative pathology in which defects in proteolytic clearance of amyloid β peptide (Aβ) likely contribute to the progressive nature of the disorder. Lysosomal proteases of the cathepsin family exhibit up-regulation in response to accumulating proteins including Aβ1–42. Here, the lysosomal modulator Z-Phe-Ala-diazomethylketone (PADK) was used to test whether proteolytic activity can be enhanced to reduce the accumulation events in AD mouse models expressing different levels of Aβ pathology. Systemic PADK injections in APPSwInd and APPswe/PS1ΔE9 mice caused 3- to 8-fold increases in cathepsin B protein levels and 3- to 10-fold increases in the enzyme's activity in lysosomal fractions, while neprilysin and insulin-degrading enzyme remained unchanged. Biochemical analyses indicated the modulation predominantly targeted the active mature forms of cathepsin B and markedly changed Rab proteins but not LAMP1, suggesting the involvement of enhanced trafficking. The modulated lysosomal system led to reductions in both Aβ immunostaining as well as Aβx-42 sandwich ELISA measures in APPSwInd mice of 10–11 months. More extensive Aβ deposition in 20-22-month APPswe/PS1ΔE9 mice was also reduced by PADK. Selective ELISAs found that a corresponding production of the less pathogenic Aβ1–38 occurs as Aβ1–42 levels decrease in the mouse models, indicating that PADK treatment leads to Aβ truncation. Associated with Aβ clearance was the elimination of behavioral and synaptic protein deficits evident in the two transgenic models. These findings indicate that pharmacologically-controlled lysosomal modulation reduces Aβ1–42 accumulation, possibly through intracellular truncation that also influences extracellular deposition, and in turn offsets the defects in synaptic composition and cognitive functions. The selective modulation promotes clearance at different levels of Aβ pathology and provides proof-of-principle for small molecule therapeutic development for AD and possibly other protein accumulation disorders.

RATIONALE The determination of the center-of-mass energy at which 50% of a precursor ion decomposes (Ecom50) during collision-induced dissociation (CID) is dependent on the chemical structure of the ion as well as the physical and electrical characteristics of the collision cell. The current study was designed to identify variables influencing Ecom50 values measured on four different mass spectrometers. METHODS Fifteen test compounds were protonated using +ve electrospray ionization and the resulting ions were fragmented across a range of collision energies by CID. Survival yield versus collision energy curves were then used to calculate Ecom50 values for each of these [M+H]+ ions on four different mass spectrometers. In addition, the relative recovery of the [M+H]+ ions of eight compounds ranging in molecular weight from 46 to 854 Da were determined at collision cell RF voltages ranging from 0 to 600 V. RESULTS Ecom50 values determined on the four instruments were highly correlated (r2 values ranged from 0.953 to 0.992). Although these overall correlations were high, we found different maximum ion recoveries depending on collision cell RF voltage. High mass ions had greater recovery at higher collision cell RF voltages, whereas low mass ions had greater recovery at lower collision cell RF voltages as well as a broader range of ion recoveries. CONCLUSIONS Ecom50 values measured on four different instruments correlated surprisingly well given the differences in electrical and physical characteristics of the collision cells. However, our results suggest caution when comparing Ecom50 values or CID spectra between instruments without correcting for the effects of RF voltage on ion transfer efficiency.

Artificial antenna complexes built via self-assembly are reported here, which indicated excellent energy transfer efficiency, macroscopic organization, unprecedented thermal stability, and ease of formation.Our system consists of four fluorescent donor-acceptor dyes, double-helical DNA and cationized bovine serum albumin, all self-assembled on cover glass slips to form functional materials. These captured radiation in the range of 330-590 nm, and excitation of any of the donor dyes resulted in efficient emission from the terminal acceptor. Excitation spectra provided unequivocal proof of energy transfer via jumper dyes, and transfer was interrupted when one of the jumper dyes was omitted, another direct evidence for cascade energy transfer. The entire assembly indicated unusually high thermal stability and continued to function efficiently even after exposure to 80 C for >169 days, an important consideration for field applications. These unusually stable, high efficiency, multichromophoric, artificial antennas are the first of their kind to demonstrate self-assembled 4-dye energy cascade, converting blue photons to red photons. † Electronic supplementary information (ESI) available: Including experimental methods, characterization of modied BSA, uorescence, circular dichroism, etc. See Scheme 1 Artificial antenna complexes constructed from donors, acceptors, cationized BSA (cBSA), and DNA.This journal is