Using amyloid autofluorescence as a biomarker for lysozyme aggregation inhibition.

The assembly of proteins into amyloidogenic aggregates underlies the onset and symptoms of several pathologies, including Alzheimer's disease, Parkinson's disease and type II diabetes. Among the efforts for fighting these diseases, there is a great demand for developing novel, fast and reliable methods for in vitro screening of new drugs that may suppress or reverse amyloidogenesis. Recent studies unravelled a progressive increase in a blue autofluorescence upon amyloid formation originated from many different proteins, including the peptide amyloid-ß, lysozyme or insulin. Herein, we propose a drug screening method using this property, avoiding the use of external probe dyes. We demonstrate that the inhibition of lysozyme amyloid formation by means of two known inhibitors, tartrazine and amaranth, can be monitored based on the autofluorescence of lysozyme amyloid aggregates. Our results show that amyloid luminescence is an intrinsic property that can be potentially applied in a screening assay, allowing the ranking of drug efficiency. The assays demonstrated here are fast to perform and suitable for scaling using microplate assays, configuring a new sensitive and economically feasible method.

Necrosis Depth and Photodynamic Threshold Dose with Redaporfin-PDT.

Predicting the extent of necrosis in photodynamic therapy (PDT) is critical to ensure that the whole tumor is treated but vital structures, such as major blood vessels in the vicinity of the tumor, are spared. The models developed for clinical planning rely on empirical parameters that change with the nature of the photosensitizer and the target tissue. This work presents an in vivo study of the necrosis in the livers of rats due to PDT with a bacteriochlorin photosensitizer named redaporfin using both frontal illumination and interstitial illumination. Various doses of light at 750 nm were delivered 15 min postintravenous administration of redaporfin. Sharp boundaries between necrotic and healthy tissues were found. Frontal illumination allowed for the determination of the photodynamic threshold dose-1.5 × 1019  photons cm-3 -which means that the regions of the tissues exposed to more than 11 mm of ROS evolved to necrosis. Interstitial illumination produced a necrotic radius of 0.7 cm for a light dose of 100 J cm-1 and a redaporfin dose of 0.75 mg kg-1 . The experimental data obtained can be used to inform and improve clinical planning with frontal and interstitial illumination protocols.

Pro-oxidant and Antioxidant Effects in Photodynamic Therapy: Cells Recognise that Not All Exogenous ROS Are Alike.

Photodynamic therapy (PDT) uses light, photosensitizer molecules and oxygen to generate reactive oxygen species (ROS) that kill cancer cells. Redaporfin, a new photosensitizer in clinical trials, generates both singlet oxygen and superoxide ions. We report the potentiation of redaporfin-PDT in combination with ascorbate and with the inhibition of antioxidant enzymes in A549 (human lung adenocarcinoma) and CT26 (mouse colon adenocarcinoma) cells. The addition of ascorbate and the inhibition of superoxide dismutase (SOD) strongly increased the phototoxicity of redaporfin towards A549 cells but not towards CT26 cells. The inhibition of catalase and the depletion of the glutathione pool also potentiate redaporfin-PDT towards A549 cells. The lower SOD activity of A549 cells might explain this difference. SOD activity levels may be explored to increase the selectivity and efficacy of PDT with photosensitizers that generate radical species.

Diffusion coefficients of sodium fluoride in aqueous solutions at 298.15 k and 310.15 k.

Mutual diffusion coefficients (interdiffusion coefficients) have been measured for sodium fluoride in water at 298.15 K and 310.15 K at concentrations between 0.003 mol dm-3 and 0.05 mol dm-3. The diffusion coefficients were measured using a conductimetric cell. The experimental mutual diffusion coefficients are discussed on the basis of the Onsager-Fuoss model. The limiting molar conductivity of the fluoride ion in these solutions at 310.15 K has been estimated using these results.