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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.
- Melanin: production from cheese bacteria, chemical characterization, and biological activitiesPublication . Ferraz, Ana Rita; Pacheco, Rita; Vaz, Pedro D.; Pintado, C. M. B. S.; Ascensão, Lia; Serralheiro, Maria LuisaPigments are compounds of importance to several industries, for instance, the food industry, where they can be used as additives, color intensifiers, and antioxidants. As the current trend around the world is shifting to the use of eco-friendly commodities, demand for natural dyes is increasing. Melanins are pigments that are produced by several microorganisms. Pseudomonas putida ESACB 191, isolated from goat cheese rind, was described as a brown pigment producer. This strain produces a brown pigment via the synthetic Mueller-Hinton Broth. This brown compound was extracted, purified, analyzed by FTIR and mass spectrometry, and identified as eumelanin. The maximum productivity was 1.57 mg/L/h. The bioactivity of eumelanin was evaluated as the capacity for scavenging free radicals (antioxidant activity), EC50 74.0 & PLUSMN; 0.2 mu g/mL, and as an acetylcholinesterase inhibitor, with IC50 575 & PLUSMN; 4 mu g/mL. This bacterial eumelanin did not show cytotoxicity towards A375, HeLa Kyoto, HepG2, or Caco2 cell lines. The effect of melanin on cholesterol absorption and drug interaction was evaluated in order to understand the interaction of melanin present in the cheese rind when ingested by consumers. However, it had no effect either on cholesterol absorption through an intestinal simulated barrier formed by the Caco2 cell line or with the drug ezetimibe.
- Phenolic profile and biological activities of decoctions from Santolina impressa, a Portuguese endemic speciesPublication . Rodrigues, Ana M.; Fale, Pedro; Madeira, Paulo; Pacheco, Rita; Florêncio, Maria Helena; Ascensão, Lia; Serralheiro, Maria LuisaSantolina impressa Hoffmanns & Link is an aromatic Asteraceae species endemic to the southwest of Portugal. It is used in folk medicine as an herbal tea for gastrointestinal ailments and mouthwash antiseptic. The present study aimed to relate the chemical composition of decoctions from the aerial parts S. impressa with their antiacetylcholinesterase, antioxidant, antimicrobial and cytotoxic activities, in an attempt to explain the traditional use of this species. LC–MS/MS analysis identified chlorogenic acid and cynarin as the main components. Antimicrobial activity of the extracts was investigated against two cariogenic bacteria:- Streptococcus sobrinus and Streptococcus mutans, and these decoctions were not cytotoxic towards Caco-2 and HepG2 cell lines. Decoctions from capitula and stems/leaves showed high AChE inhibition, with IC50 values of 328 μg/mL and 579 μg/mL, respectively. The antioxidant activity measured using the DPPH radical scavenging assay showed an EC50 of 14.7 μg/mL and 12.9 μg/mL for capitula and stems/leaves extracts, respectively. The biological activities were kept constant after in vitro digestive process and the HPLC analysis did not indicate changes in the extract’s chemical composition. The results may explain the traditional use of S. impressa decoctions for digestive problems and as a mouthwash antiseptic.
- Nanoformulation of seaweed eisenia bicyclis in albumin nanoparticles targeting cardiovascular diseases: In vitro and in vivo evaluationPublication . Pinto, Sofia; Gaspar, Maria Manuela; Ascensão, Lia; Faísca, Pedro; Reis, Catarina; Pacheco, RitaNatural products, especially those derived from seaweeds, are starting to be seen as effective against various diseases, such as cardiovascular diseases (CVDs). This study aimed to design a novel oral formulation of bovine albumin serum nanoparticles (BSA NPs) loaded with an extract of Eisenia bicyclis and to validate its beneficial health effects, particularly targeting hypercholesterolemia and CVD prevention. Small and well-defined BSA NPs loaded with Eisenia bicyclis extract were successfully prepared exhibiting high encapsulation efficiency. Antioxidant activity and cholesterol biosynthesis enzyme 3-hydroxy-3 methylutaryl coenzyme A reductase (HMGR) inhibition, as well as reduction of cholesterol permeation in intestinal lining model cells, were assessed for the extract both in free and nanoformulated forms. The nanoformulation was more efficient than the free extract, particularly in terms of HMGR inhibition and cholesterol permeation reduction. In vitro cytotoxicity and in vivo assays in Wistar rats were performed to evaluate its safety and overall effects on metabolism. The results demonstrated that the Eisenia bicyclis extract and BSA NPs were not cytotoxic against human intestinal Caco-2 and liver HepG2 cells and were also safe after oral administration in the rat model. In addition, an innovative approach was adopted to compare the metabolomic profile of the serum from the animals involved in the in vivo assay, which showed the extract and nanoformulation's impact on CVD-associated key metabolites. Altogether, these preliminary results revealed that the seaweed extract and the nanoformulation may constitute an alternative natural dosage form which is safe and simple to produce, capable of reducing cholesterol levels, and consequently helpful in preventing hypercholesterolemia, the main risk factor of CVDs.