Protective effect of Pinot noir pomace extract against the cytotoxicity induced by polycyclic aromatic hydrocarbons on endothelial cells.

Polycyclic Aromatic Hydrocarbons (PAHs) are pollutants found in the air generated mainly by the combustion of coal or biomass burning. Exposure to Polycyclic Aromatic Hydrocarbons is positively correlated with cardiovascular diseases. Phenolic compounds are widely found in the plant kingdom, and their availability from agri-food processing waste has led to an increased interest in their recovery. The production of large amounts of organic waste created by the wine industry has emphasized the valuation of these wastes to generate high-added-value by-products. The objective of this work was to investigate the protective effect of Pinot noir pomace extract on human endothelial cells against PAHs found in the polluted air of Temuco, Chile. The pomace extract was characterized by spectrophotometric analysis and high-performance liquid chromatography (HPLC). Results revealed the presence of 5 glycosylated anthocyanins and 9 low molecular weight polyphenols. Molecular docking indicated that cyanidin-3-glucoside (-9.2 kcal/mol) and quercetin (-9.6 kcal/mol) had the highest affinities for the Nrf2 binding site in the Keap1 protein, suggesting a possible competition with this transcription factor. Endothelial cells from the human umbilical vein were exposed to increasing concentrations of Phenanthrene, Fluoranthene, and Pyrene diluted in DMSO in a ratio of 3:1:1 (10 µM-200 µM). Viability through the MTS assay showed that 150 µM of PAHs was sufficient to reduce viability by 75% (p ˂ 0.0001). When the cells were pre-treated with 400 µg/ml of the extract, 150 µM of PAHs did not exert cell death (80% viability). Our preliminary results show that polyphenolic components found in Pinot noir pomace might have a beneficial effect as a protective agent.

From Synthesis to Characterization of Site-Selective PEGylated Proteins.

Covalent attachment of therapeutic proteins to polyethylene glycol (PEG) is widely used for the improvement of its pharmacokinetic and pharmacological properties, as well as the reduction in reactogenicity and related side effects. This technique named PEGylation has been successfully employed in several approved drugs to treat various diseases, even cancer. Some methods have been developed to obtain PEGylated proteins, both in multiple protein sites or in a selected amino acid residue. This review focuses mainly on traditional and novel examples of chemical and enzymatic methods for site-selective PEGylation, emphasizing in N-terminal PEGylation, that make it possible to obtain products with a high degree of homogeneity and preserve bioactivity. In addition, the main assay methods that can be applied for the characterization of PEGylated molecules in complex biological samples are also summarized in this paper.