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- Effect of food preparations on in vitro bioactivities and chemical components of fucus vesiculosusPublication . André, Rebeca; Guedes, Laura; Melo, Ricardo; Ascensão, Lia; Pacheco, Rita; Vaz, Pedro D.; Serralheiro, Maria LuisaFucus vesiculosus is a brown macroalgae used in food and generally considered safe to be consumed, according to EU Directive (EC 258/97). The aim of this study is to analyze the e_ect of food preparation on F.vesiculosus of di_erent origins on what concerns its chemical constituents and final bioactivities. The aqueous extract of the seaweeds were obtained at di_erent temperatures, similar to food preparation and then purified by SPE. The compound identification was carried out by Liquid Chromatography High Resolution Mass Spectrometry (LC-HRMS/MS) and algae extracts microstructure were observed by Scanning Electron Microscopy (SEM). The activities were determined by using antioxidant activity, inhibition of acetylcholinesterase (AChE) and 3-hidroxi-3-methyl-glutaril-CoA (HMG-CoA) reductase (HMGR) together with Caco-2 cells line simulating the intestinal barrier. The activity of AChE and the HMGR were inhibited by the extracts giving IC50 values of 15.0 _ 0.1 _g/mL and 4.2 _ 0.1 _g/mL, respectively and 45% of the cholesterol permeation inhibition. The main compounds identified were phlorotannins and peptides derivatives. The mode of preparation significantly influenced the final bioactivities. Moreover, the in vitro results suggest that the preparation of F. vesiculosus as a soup could have hypercholesterolemia lowering effect.
- Action of euptox A from Ageratina adenophora juice on human cell lines: a top-down study using FTIR spectroscopy and protein profilingPublication . André, Rebeca; Catarro, Joana; Freitas, Dalia; Pacheco, Rita; Oliveira, Maria Conceição; Serralheiro, Maria Luisa; Fale, PedroEuptox A, from Ageratina adenophora juice, is a toxin associated with the plant's resistance to infections, invasiveness and traditional use in cancer treatment. We used FTIR spectroscopy and protein profiling of cell lines to study the impact of euptox A on human cells, to clarify its mechanism of action in a top-down approach. Euptox A was extracted from the juice of A. adenophora. Its stability in the gastrointestinal tract was evaluated, as the compound/juice is generally taken orally. Cytotoxicity was determined in HeLa, Caco-2 and MCF7 cells, and the mechanism of action analyzed by protein and metabolite profiles using electrophoresis and FTIR spectroscopy. Euptox A resisted gastrointestinal digestion and was the most cytotoxic component of the extract for all cell lines tests. Euptox A-treated HeLa cells showed changes in protein profile, especially on 40S ribosomal protein S8 (RP), generally associated with cancer cells. FTIR profiles of treated cells diverged in the same metabolites as cells treated with cisplatin, both in metabolite directed analysis and in multivariate analysis (principal component analysis). In conclusion, euptox A in this top-down study showed a cellular impact that suggests a strong potential against cancer, acting on cancer targeted cellular characteristics.
- Brown algae potential as a functional food against hypercholesterolemia: reviewPublication . André, Rebeca; Pacheco, Rita; Bourbon, Mafalda; Serralheiro, Maria LuisaBrown algae have been part of the human diet for hundreds of years, however, in recent years, commercial and scientific interest in brown algae has increased due to the growing demand for healthier diet by the world population. Brown algae and its metabolites, such as carotenoids, polysaccharides, phlorotannins, and proteins, have been associated with multiple beneficial health effects for different diseases, such as cardiovascular diseases, one of the main causes of death in Europe. Since high blood cholesterol levels are one of the major cardiovascular risks, this review intends to provide an overview of current knowledge about the anti-hypercholesterolemic effect of different brown algae species and/or their isolated compounds.
- Bioactivities of iridoids and flavonoids present in decoctions from aerial parts of Verbascum betonicifoliumPublication . Fadel, Sezan R.; Bendif, Hamdi; Guedes, Laura; André, Rebeca; Pacheco, Rita; Guedes, Rita; Merabti, Karim; Miara, Mohamed Djamel; Serralheiro, Maria LuisaIntroduction: Verbascum betonicifolium (V. betonicifolium) is a plant used in traditional medicine for several ailments. The objective of this study was to determine the antioxidant activity and acetylcholinesterase inhibitory activity of the aqueous extract together with the metabolites responsible for these activities. This paper presents the first phytochemical characterization and bioactivities of aqueous extracts from aerial parts. Methods: The compounds present in the aerial part aqueous extract were identified by high performance liquid chromatography, coupled to a diode-array detector (HPLC-DAD) and by high-resolution mass spectrometry (HRMS), using LC-MS/MS analyses. Antioxidant activity was measured as the capacity to scavenge the free radical DPPH and the AChE activity was determined using the Ellman test. The cytotoxicity was determined used HepG2 cell lines. Results: Several types of metabolites were found, primary metabolites (malic, citric and gluconic acids), phenolic compounds (verbascoside, luteolin), terpenoids (iridoid glycoside, unedide), among others. The total phenol content of 28 mu g of gallic acid equivalents/mg of extract was determined. The aqueous extract antioxidant activity had and EC50 of 70 mu g/mL and the AChE inhibitory activity an IC50 of 750 mu g/mL. No cytotoxicity towards HepG2 cells was detected, even using a concentration of 1 mg/mL. Conclusions: The phenolic compounds present in the extract may be the main contributors to the bioactivities of V. betonicifolium. These results show for the first time the richness of phytochemicals and the strong bioactivities of V. betonicifolium and that the aqueous extract could be used as new natural sources of bioactive molecules.