HRP
Melanin, a pigment synthesized by melanocytes, serves as the primary defense against UV-induced skin damage due to its potent antioxidant properties. There is increasing interest in natural substances capable of modulating the melanogenic pathway, particularly in hypopigmentation disorders. This study investigated the effect of methanolic extracts from the diatom Odontella aurita-authorized as a food supplement in the EU-on melanogenesis in the B16-F0 murine melanoma cell line. The research evaluated melanin content, tyrosinase activity, and the expression of melanogenesis-related genes and proteins at defined time points. Metabolomic and biochemical analyses were performed to characterize the extract's composition. Treatment with O. aurita extract significantly increased melanin content in B16-F0 cells by 45% (p < 0.01) compared to control. Tyrosinase activity was elevated by 38% after 24 h (p < 0.01), with gene and protein expression analyses confirming upregulation of Tyrosinase (TYR) after 0.5 h, Tyrosinase Related Protein-1 (TRP1) after 1 h, and Tyrosinase Related Protein-2 (TRP2) after 8 h. The extract also enhanced the cellular antioxidant environment, as evidenced by increased levels of metabolic cofactors and pigment-precursor amino acids. O. aurita methanolic extract accelerates and sustains melanin synthesis and tyrosinase activity, distinguishing its effect from single-compound inducers. These findings support the therapeutic potential of O. aurita for pigmentary disorders and skin health. Further studies should investigate its efficacy and safety in vivo and explore its application in cosmeceutical and nutraceutical formulations.
Glycolate oxidase (HAO1) catalyses the synthesis of glyoxylate, a common metabolic intermediate that causes renal failure if accumulated. HAO1 inhibition is an emerging treatment for primary hyperoxaluria, a rare disorder of glyoxylate metabolism. Here we report the first cell-based measurement of inhibitor uptake and engagement with HAO1, by adapting the cellular thermal shift assay (CETSA) based on Nano luciferase complementation and luminescence readout. By profiling the interaction between HAO1 and four well-characterised inhibitors in intact and lysed HEK293T cells, we showed that our CETSA method differentiates between low-permeability/high-engagement and high-permeability/low-engagement ligands and is able to rank HAO1 inhibitors in line with both recombinant protein methods and previously reported indirect cellular assays. Our methodology addresses the unmet need for a robust, sensitive, and scalable cellular assay to guide HAO1 inhibitor development and, in broader terms, can be rapidly adapted for other targets to simultaneously monitor compound affinity and cellular permeability.
