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Euterpe edulis Martius, popularly known as Juçara, is a native tree of the Atlantic Rainforest found predominantly in the states of the southern and southeastern regions of Brazil. Juçara fruit is a globose berry that, when ripe, acquires a dark purple shade and sweet pleasant taste. The industrial production of Juçara fruit pulp generates solid residues (peel) which are usually discarded [1]. The aim of the present work was to perform an unprecedented in-depth study on the bioactive components profile of E. edulis fruit peel. The nutritional composition of this material was estimated and its hydroethanolic extract was characterized in terms of phenolic compounds, besides antioxidant and antibacterial potential. Finally, the hepatotoxicity of the extract was assessed. A total of nineteen phenolic compounds was identified in Juçara peel. Seventeen were non-anthocyanin molecules: two phenolic acids, namely caffeic acid and a ferulic acid derivative; four flavanonols, corresponding to three dihydroquercetin (taxifolin) and one dihydrokaempferol (aromadendrin) glycoside derivatives; six flavones, assigned as apigenin C-glycoside derivatives; and five flavonols, among which quercetin-3-O-rutinoside, kaempferol-3-O-rutinoside, and isorhamnetin-3-O-rutinoside. The major components of the peel extract were the anthocyanins cyanidin-3-O-rutinoside and cyanidin-3-O-glucoside (5.32 and 6.23 mg/g of extract, respectively), which together accounted for more than 87% of the extract’s total phenolic content, which corroborates literature data [2, 3]. Anthocyanins are amidst the main compounds related to the great free radical-scavenging capacity of Juçara fruit, whereas a significant positive correlation with its general antioxidant capacity was observed (Shultz et al., 2016). The herein studied fruit peel extract presented expressive values of antioxidant capacity, assessed by five distinct methods, namely (1) oxidative haemolysis inhibition assay (OxHLIA), (2) inhibition of the production of thiobarbituric acid reactive substances (TBARS); (3) reduction of the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH); (4) reduction of the 2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonate) cation (ABTS), and (5) reduction power of the ferric ion (FRAP). Furthermore, the results obtained for antioxidant activity were more expressive than the ones verified by other authors for Juçara residues [1]. The evaluated E. edulis extract showed no toxicity against a non-tumour liver primary culture PLP2, at the highest concentration assessed (400 μg/mL). The inhibitory activity displayed by the extract against both Gram-positive (Enterococcus faecalis, Listeria monocytogenes, and Methicillin-resistant Staphylococcus aureus - MRSA) and Gram-negative (Escherichia coli, Klebsiella pneumonia, Morganella morganii, and Pseudomonas aeruginosa) bacteria indicates the presence of an extensive spectrum of phytochemical constituents with antibiotic potential. Indeed, the extract was more effective than the antibiotic ampicillin against M. morganii and P. aeruginosa. Therefore, Juçara fruit residues could be used to produce high added-value food ingredients, both colorants and preservatives, following the circular bioeconomy concept and stimulating the Juçara production chain. R.C.G. Corrêa (Process number 167378/2017-1), R.M. Peralta (Project number 307944/2015-8) and A. Bracht (Project number 304090/2016-6) thank Conselho Nacional de Desenvolvimento Científico e Tecnologia (CNPq), Brazil, for the financial support. The authors are grateful to FCT - Portugal and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019), L. Barros, R. C. Calhelha and C. Pereira contracts; to FEDER-Interreg España-Portugal for financial support through the project 0377_Iberphenol_6_E; FEDER through the Regional Operational Program North 2020: Project NORTE-01-0145-FEDER-023289 (DeCodE) and project Mobilizador Norte-01-0247-FEDER-024479 (ValorNatural®). info:eu-repo/semantics/publishedVersion

Plant-derived compounds have been extensively used for cosmeceutical applications, especially because humans have once again turned to Nature to mitigate the relative void of combinatorial chemistry, to find new molecules and the toxicological effects associated with the synthetic ones [1]. Cytinus hypocistis (L.) L. is a wild edible parasitic plant on various members of the Cistaceae family. Although its biological properties were potentially attributed to its hydrolysable tannins content, to the author's best knowledge, its chemical composition is largely unknown, and active biomolecules are not yet identified [2]. According to a semi-quantitative study, where 100 extracts obtained from plants collected in India, Africa, and the Mediterranean area, C. hypocistis figures on the top 10 group of plants that potently inhibited both elastase and tyrosinase, two main enzymes involved in skin aging [3]. Thus, studying the bioative properties and chemical composition of C. hypocistis plant will give comprehensive clues on its potential cosmeceutical applications. Plant specimens of C. hypocistis were collected in June 2018 in Castro Daire, Portugal. After lyophilisation, the phenolic compounds were analysed in the hydroethanolic extracts of four different parts of C. hypocistis (whole plant, nectar chamber of the flower, petals, and stalks) using a HPLC-DAD-ESI/MSn system. The antioxidant activity of the four extracts were evaluated using OxHLIA and TBARS methodology. Anti-tyrosinase enzyme inhibitory assay was performed using L-DOPA as substract and kojic acid as standard. C. hypocistis extracts were also tested for their antibacterial activity based on minimum inhibitory and bactericidal concentrations and the anti-inflammatory activity was evaluated through NO inhibition, in LPS-activated murine macrophage (RAW 264.7). A total of 17 phenolic compounds were identified, being galloyl-bis-HHDP-glucose, digalloyl-bis-HHDP-glucopyranose, and pedunculagin the most abundant. UV radiation generates oxidative stress, being mainly responsible for cell membrane oxidation and, although through different mechanisms, OxHLIA and TBARS are equally a consequence of lipid peroxidation. All the tested extracts showed high antioxidant capacity, with the petals exhibiting the most promising results for both OxHLIA (IC50 = 279 ± 5 ng/mL) and TBARS (IC50 = 342 ± 2 ng/mL) assays. Considering the anti-tyrosinase inhibitory assay, the main enzyme involved in skin pigmentation, the stalks presented the lowest IC50 values, 0.09 ± 0.02 mg/mL. All tested extracts displayed a broad-spectrum microbial inhibition against both Gram-positive and Gram- negative bacteria. Moreover, being chronic inflammation one of the molecular mechanisms behind skin aging, the petals result for NO inhibition (IC50: 127 ± 8 μg/mL) is an important evidence on the versatile profile of this plant. Although for the four studied samples the 17 identified phenolic compounds were the same, its concentration was higher in the petals extract, followed by the stalks, being these two plant parts of C. hypocistis unveiling the strongest bioactive potential. These results point a potential correlation between the phenolic profile of C. hypocistis and its properties. For its bioactivity validation and mechanism investigation, further studies on fractionation, isolation and characterization of compounds of the extracts of C. hypocistis are currently ongoing. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019). L. Barros, J. Pinela, M.I. Dias and R.C. Calhelha thank the national funding by FCT, P.I., through the institutional scientific employment program-contract for their contracts. Funding: The authors are also grateful to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion

Plants are a considerable source of natural products and are often used by man for their aromatic, medicinal and flavouring abilities. Aromatic herbs, in particular, have been used for centuries in gastronomy but also in traditional medicine. Considering that a growing number of consumer’s associate products containing natural ingredients to a higher quality, the food industry is also giving a greater importance to the possible use of plant extracts and/or essential oils as alternatives to synthetic additives in foods. Lovage (Levisticum officinale W.D.J. Koch) is an aromatic plant used as a condiment in several regions of Europe that is also used as an ingredient of commercial food seasonings. So far, studies on lovage mainly focused on its essential oil composition, but there is still a scarcity of information regarding its nutritional and chemical composition as well as its bioactive properties. To help start bridging this gap, and to explore alternative uses of this plant species, the present work reports a comprehensive study on lovage composition and bioactivity. Fresh plant specimens were commercially acquired in October 2018 at Porto, Portugal. After lyophilization, the aerial parts of the plant were further analyzed. The proximate composition (moisture, fat, proteins, ash, carbohydrates and energy) was evaluated by AOAC official procedures, free sugars and tocopherols were determined using liquid chromatography coupled to a refraction index (HPLC-RI) and fluorescence (HPLC-FL) detectors, respectively. Fatty acids were determined by gas chromatography coupled to a flame ionization detector (GC-FID), organic acids by ultra-fast liquid chromatography coupled to a diode detector (UPLC-DAD) and phenolic compounds by HPLC coupled to a DAD and mass spectrometry (MS) using the electrospray ionization interface (ESI). Multi-resistant clinical bacterial strains were used to screen the antimicrobial activity of aqueous (decoction) and hydroethanolic extracts. The antioxidant activity was evaluated through five different in vitro assays: DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity, reducing power, inhibition of β-carotene bleaching, inhibition of lipid peroxidation in brain homogenates by thiobarbituric acid reactive substances (TBARS) and oxidative hemolysis inhibition assay (OxHLIA). The cytotoxic activity was assayed using MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung carcinoma), HeLa (cervical carcinoma) and HepG2 (hepatocellular carcinoma) human tumor cell lines, and also a non-tumor cell line (porcine liver primary cells, PLP2). L. officinale presented a high percentage of moisture with proteins being the predominant macronutrient. In what concerns the chemical composition of the plant, lovage showed a predominance of polyunsaturated fatty acids with alpha-linolenic acid being the major compound; alpha tocopherol stands out among the other isoforms of vitamin E; oxalic acid has a prominence among the identified organic acids, and glucose was the main free sugar present in the plant. Regarding phenolic compounds’ composition, a total of 7 compounds, including phenolic acids and flavonoids, were identified and quantified, with 5-O-caffeoylquinic acid being the predominant one. Both extracts presented interesting antioxidant properties, and in general, were more active against Gram-negative bacteria, while only decoction extract showed cytotoxicity against a HepG2 cell line. In brief, an extensive and detailed study was performed revealing that lovage has an interesting composition from the nutritional point of view being also a source of several bioactive compounds, therefore its inclusion as a seasoning/flavoring agent in different dishes should be promoted. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019), L. Barros and A. Fernandes contracts; and to FEDER-Interreg España-Portugal program info:eu-repo/semantics/publishedVersion

Myrciaria cauliflora (Mart.) O. Berg, known as jabuticaba, is a native species to Brazil, more specifically to Atlantic Rainforest biome. Its fruits are small berries with a diameter of 2.0 – 3.5 cm that contain between one and four seeds, with a sweet gelatinous pulp and thick dark purple epicarp when mature, which is not commonly consumed. Due to the properties of its pulp, the consumption and production of jellies and liqueurs from jabuticaba has increased. However, the epicarp that corresponds to about 50% of the fruit is not used, being converted into a bioresidue [1]. The intense colour of this part of the fruit is due to the pigments, namely anthocyanins, present in its composition. In addition to attractive staining, these molecules have been associated with bioactive properties, such as antioxidant activity, which makes their recovery interesting for application in various industrial segments, for instance, food and pharmaceutical [2,3]. With the objective of valorising this bioresidue, the present work aimed the determination of the main anthocyanins present in jabuticaba epicarp (by HPLC-DAD-ESI/MS) and the evaluation of the bioactivity of its ethanolic extract (through in vitro assessment of cytotoxic, antimicrobial, and antioxidant activities). Thus, cytotoxicity was evaluated in four tumour cell lines (NCI-H460 - lung carcinoma, MCF-7 - breast carcinoma, HepG2 - hepatocellular carcinoma, and HeLa - cervical carcinoma) and in a primary culture of non-tumour liver cells (PLP2), by the Sulforodamine B (SRB) assay. The antimicrobial activity was evaluated on five Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Morganella morgani, Proteus mirabilis, and Pseudomonas aeruginosa) and three Gram-positive bacteria (Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis). Lastly, the antioxidant activity was tested through the oxidative haemolysis inhibition assay (OxHLIA). The bioresidue of jabuticaba fruits presented two anthocyanins, identified as cyanidin 3-O-glucoside and delphinidin 3-O-glucoside, being the first one the most abundant. Regarding bioactivity, the ethanolic extract revealed antiproliferative activity in all tumour cell lines evaluated (GI50 < 300 μg/mL), except for NCI-H460, and did not show toxicity for PLP2 at the maximal tested concentration (400 μg/mL). It also exhibited bacteriostatic properties in all the analysed bacterial strains (ranging from 20 to 10 mg/mL). Regarding haemolysis inhibition, the extract was able to protect 50% of the erythrocyte population for 120 minutes in a lower concentration than the positive control (Trolox). The results obtained in this study allow to conclude that jabuticaba epicarp is a rich source of anthocyanins and also exhibits strong bioactivity, which makes it suitable for use as colorant. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019), L. Barros, R.C. Calhelha, and C. Pereira contracts and B. Albuquerque research grant (SFRH/BD/136370/2018). This work was also funded by the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project NORTE-01-0145-FEDER-023289: DeCodE and project Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®.The authors are grateful to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion

A jabuticabeira é uma árvore frutífera nativa do Brasil, sendo encontrada principalmente na Mata Atlântica. Os seus frutos, denominados por jabuticabas, são pequenas bagas de aproximadamente 2 – 3,5 cm que possuem uma coloração preta a roxa, quando maduros. São muito apreciados para consumo in natura devido ao agradável sabor da sua polpa; no entanto, o epicarpo, que é rígido e possui um sabor adstringente, não é comumente consumido, sendo responsável por grande parte do resíduo gerado no processamento de produtos derivados de jabuticaba [1]. A intensa coloração desta parte dos frutos é devida à concentração de pigmentos presentes na sua composição, nomeadamente antocianinas, que são compostos de grande interesse devido à sua coloração atrativa e às suas propriedades benéficas para a saúde [2]. Além disso, a presença de outros compostos fenólicos também tem sido relatada no epicarpo de jabuticaba [1,3]. Com o objetivo de valorizar este biorresíduo, o presente estudo teve como foco a determinação do seu perfil fenólico e a avaliação das suas propriedades bioativas, nomeadamente a atividade antioxidante, antimicrobiana e antiproliferativa. Os compostos fenólicos presentes no extrato do epicarpo de jabuticaba (EEJ) foram detetados por HPLC-DAD- ESI/MS. Os ensaios de bioatividade foram realizados por métodos in vitro: a atividade antioxidante foi determinada através da inibição da formação de substâncias reativas do ácido tiobarbitúrico (TBARS) e da inibição da hemólise oxidativa (OxLHIA); a atividade antimicrobiana foi avaliada pelo método da microdiluição contra oito bactérias patogénicas; e a citotoxicidade foi analisada pelo ensaio da sulforrodamina B com quatro linhas celulares tumorais e uma cultura primária de células não-tumorais. O epicarpo de jabuticaba apresentou um perfil fenólico constituído por catorze compostos fenólicos não antociânicos e dois antociânicos, sendo a cianidina-3-O-glucósido o composto mais abundante. Em relação às propriedades bioativas, o EEJ apresentou uma elevada atividade antioxidante, atividade antiproliferativa contra três linhas celulares tumorais e atividade antimicrobiana contra todas as bactérias avaliadas. Não foi observada hepatotoxicidade em células não-tumorais na maior concentração de extrato avaliada. Conclui-se, assim, que o EEJ apresenta uma composição rica em compostos fenólicos, principalmente em antocianinas, e que possui consideráveis propriedades bioativas, o que potencia o interesse da exploração deste biorresíduo para a obtenção de compostos de interesse para a indústria alimentar e farmacêutica. Os autores agradecem à Fundação para a Ciência e a Tecnologia (FCT, Portugal) e ao FEDER no âmbito do programa PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2019); financiamento nacional da FCT, PI, através do contrato de programa institucional de emprego científico para o contrato de L. Barros e R.C. Calhelha; o contrato de C. Pereira através da celebração contrato de programa previso no no 4, 5 e 6 do artigo 23o do Decreto-Lei no 57/2016, de 29 de agosto, alterado pela Lei no 57/2017, de 19 de julho; e bolsas de doutoramento de B. Albuquerque (SFRH/BD/136370/2018) e T.C.S.P. Pires (SFRH/BD/129551/2017); FEDER através do Programa Operacional Regional Norte 2020, no âmbito do Projeto NORTE-01-0145-FEDER-023289 (DeCodE) e projeto Mobilizador Norte-01- 0247-FEDER-024479: ValorNatural®; FEDER-Interreg España-Portugal pelo apoio financeiro através do projeto 0377_Iberphenol_6_E e TRANSCoLAB 0612_TRANS_CO_LAB_2_P. info:eu-repo/semantics/publishedVersion

Punica granatum L. (pomegranate), is a fruit that has been widely studied due to its composition in specific phenolic compounds, having also been described as displaying excellent bioactive properties (antioxidant, antimicrobial, and anti-inflammatory activity), with preventive action against some diseases [1,2]. Given that the inedible part of this fruit can represent up to about 50% of the total weight, this bio-waste has high potential as a source of bioactive compounds of interest [3]. Thus, this study compared different types of solvents in the extraction of phenolic compounds from the exocarp, mesocarp and pericarp of two pomegranate varieties: Mollar de Elche (yellow pomegranate) and Purple Queen (red pomegranate). Chemical characterization was performed by HPLC-DAD-ESI/MS and revealed the presence of fourteen phenolic compounds in the Mollar de Elche, highlighting punicalagin and galloyl-HHDP-glucose as the major compounds. The extracts of pomegranate Purple Queen showed the same 14 phenolic compounds, despite being present in lower amounts, but this cultivar revealed also three anthocyanins, which may justify the characteristic reddish color of its bark. Additionally, the antioxidant activity (inhibition of thiobarbituric acid reactive species formation in brain homogenates and inhibition of oxidative hemolysis, OxHLIA), antibacterial (against a panel of 5 Gram-positive bacteria and 3 Gram-negative bacteria, using the microdilution method), anti-inflammatory activity (rat macrophage cells), and cytotoxic potential (in human tumor cell lines by the sulforhodamine B method). Mollar de Elche stood out for its higher antioxidant and antibacterial activity, and cytotoxicity. In general, the different extracts obtained from the by-products of both pomegranate varieties represent a source of phenolic compounds that allow it to exhibit excellent bioactive properties with potential application in the food industry as a natural ingredient, which will allow valuing these bio-residues. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019); national funding by FCT, P.I., through the institutional scientific employment program-contract for L. Barros and R. Calhelha’s contract, though the celebration of program-contract foreseen in No. 4, 5 and 6 of article 23o of Decree-Law No. 57/2016, of 29th August, amended by Law No. 57/2017, of 19th July; research contract of C. Caleja (Project AllNatt, POCI- 01-0145-FEDER-030463); to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. This work is funded by the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project NORTE-01-0145-FEDER-023289: DeCodE and project Mobilizador Norte- 01-0247-FEDER-024479: ValorNatural®. info:eu-repo/semantics/publishedVersion

Topical formulations are designed to exert multifunctional benefits to the skin, and the interest on natural bioactive compounds as cosmeceutical ingredients have recently received increased attention [1]. However, the utilization of natural ingredients can present constraints related to their stability (e.g. against pH and temperature), being microencapsulation a useful strategy to overcome some of these limitations [2]. The present work describes p-hydroxybenzoic acids anti-inflammatory, anti-tyrosinase, and antimicrobial activity. The compound was further microencapsulated using the atomization/coagulation method, with sodium alginate coagulated with calcium chloride. The obtained microspheres were characterized in terms of morphology, particle size distribution, and encapsulation efficiency. Free and microencapsulated forms were then incorporated into a semisolid cosmetic base formulation and HPLC-DAD was used to screen the presence of the compound in the final formulation. p-Hydroxybenzoic acid showed anti-inflammatory (EC50 = 195 ± 9 μg/mL), anti-tyrosinase (EC50 = 1.86 ± 0.01 mg/mL), and antimicrobial activity against a panel of Gram positive and Gram negative bacteria, and also Candida albicans. The microparticles showed spherical morphology, with various sizes (D10, D50 and D90 representing 20.5, 74.6, and 206 μm, respectively), little agglomeration, and a unimodal and bimodal particle size distribution (number and volume, respectively). Encapsulation efficiency and yield were 88.3% and 57.8%, respectively. After incorporation, the formulation containing free forms of the compound maintained some of its bioactive properties, while the encapsulated forms preserved the bioactivity showing a slow release profile of the compounds. In conclusion, the studied compound showed interesting bioactive properties and can be used in the development of multifunctional cosmeceutical formulation. The adopted encapsulation strategy also provides a suitable alternative to prolong retention of bioactive compounds for subsequent release (sustained release), thereby preserving its bioactivity over time. FCT and FEDER under Program PT2020 (POCI-COMPETE2020) for CIMO (UID/AGR/00690/2019) and Associate Laboratory LSRE-LCM (UID/EQU/50020/2019) funded by national funds through FCT/MCTES (PIDDAC). European Structural and Investment Funds (FEEI) through NORTE 2020 (Project Mobilizador ValorNatural®). This work is funded by the European Agricultural Fund for Rural Development (EAFRD), through the Rural Development Program (PDR2020), within the scope of Project MicoCoating (PDR2020-101-031472). L. Barros. R.M. Calhelha and I. Fernandes thank the national funding by FCT, P.I., through the institutional scientific employment program-contract for their contracts, while S. Heleno thanks the national funding by FCT, P.I., through the individual scientific employment program-contract. info:eu-repo/semantics/publishedVersion

Ocimum basilicum var. purpurascens (vulgarmente conhecido como manjericão vermelho rubi) é uma planta anual, pertencente à família das Lamiaceae, apresentando uma cor púrpura devido à presença de compostos antociânicos. Estas moléculas apresentam um elevado interesse industrial, devido às suas propriedades corantes e bioativas (atividade antioxidante e antimicrobiana). Com a finalidade de maximizar a extração destes compostos a partir de matrizes naturais, aplicam-se modelos matemáticos, nomeadamente a metodologia de superfície de resposta (RSM), onde se avaliam as respostas do método de extração a diferentes variáveis1. O objetivo deste trabalho consistiu em otimizar o processo de extração de compostos antociânicos, particularmente derivados de cianidina e pelargonidina, recorrendo ao RSM, de forma a promover a utilização destes pigmentos a nível industrial. As folhas do manjericão vermelho rubi foram submetidas a extrações assistidas por calor, a fim de obter um extrato rico em compostos antociânicos. Para se otimizar o processo de extração foi aplicado um desenho composto central circunscrito (DCCC) com 5 níveis para cada uma das variáveis avaliadas. Assim, as variáveis estudadas foram: tempo de extração (t, 20-120 min), temperatura (T, 25-85 ºC) e solvente (S, 0-100 % de etanol, v/v). A determinação do perfil antociânico foi realizada através de um sistema de HPLC-DAD-ESI/MS e a quantificação destes compostos nos extratos foi utilizada como variável de resposta. A avaliação das propriedades bioativas do extrato obtido, foi efetuada através de ensaios de citotoxicidade, utilizando linhas celulares tumorais (HepG2, NCI-H460, MCF-7 e HeLa) e uma cultura de células primárias não tumorais (PLP2), e de avaliação da atividade antimicrobiana (utilizando um painel de bactérias Gram-positivo e Gram-negativo, e fungos). As condições ótimas de extração dos compostos antociânicos foram: t= 65,37 ± 3,62 min, T= 85,00 ± 1,17 ºC e S=62,50 ± 4,24 %, originando um extrato rico nestas moléculas com 114,74 ± 0,58 mg de antocianinas por g de extrato. Relativamente às propriedades bioativas do extrato otimizado, este revelou inibição do crescimento celular nas linhas HeLa (GI50= 213 ± 9 μg/mL) e HepG2 (GI50= 198 ± 9 μg/mL) e ausência de citoxicidade na linha PLP2 (GI50 >400 μg/mL). Na atividade antimicrobiana, os resultados obtidos foram satisfatórios para a maioria das bactérias e fungos testados, apresentando valores de CMI (concentração mínima inibitória) entre 0,75 - 0,075 mg/mL, para Salmonella typhimirium e Listeria monocytogenes/Escherichia coli, respetivamente. Os resultados obtidos neste estudo permitem concluir que as folhas do manjericão vermelho rubi são uma matriz promissora para a extração de compostos corantes e bioativos, utilizando solventes verdes e tempos de extração reduzidos, suscitando elevado interesse ao nível de vários setores industriais, tais como o sector farmacêutico e alimentar. FCT (Portugal) e ao FEDER no âmbito do programa PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2019); fundos nacionais através da FCT, I.P., no âmbito da celebração do contrato-programa de emprego científico institucional pelos contratos de L. Barros e R.C Calhelha; FEDER-Interreg España-Portugal pelo apoio financeiro através do projeto 0377_Iberphenol_6_E; FEDER através do Programa Operacional Regional Norte 2020, no âmbito do Projeto NORTE-01-0145-FEDER-023289 (DeCodE) e do Projeto Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®. info:eu-repo/semantics/publishedVersion

Extrusion-cooking technology is a high-temperature and short-time process, necessary to cause structural, physico-chemical and nutritional changes of raw materials, forcing the material to flow under different conditions (temperature, moisture, screw speed, and feed). Rice and legumes have a great potential in the development of healthier gluten-free products than traditional snacks [1]. Rice flour (Oryza Sativa L.) has been reported as a good raw material to obtain expanded food products [1]. Dry beans (Phaseolus vulgaris L.) are rich in proteins, dietary fibre, complex carbohydrates (leading to low glycaemic index), minerals and numerous phytochemicals endowed with useful biological activities [2]. Carob fruit (Ceratonia siliqua L.) is a tree leguminous native to the Mediterranean region. The fortification with carob fruit would provide adequate fibre content and a good amount of bioactive compounds. In previous studies, carob evidenced therapeutical properties against several diseases, such us, regulatory effect in blood glucose level, reduction in low-density-lipoprotein cholesterol in hypercholesteraemic patients, body weight benefits, and improved digestion, and lipid utilization [3]. In this study, novel gluten-free extruded foods (composed by rice: 50-80%, bean: 20-40%, and carob: 0-10%) were performed using a twin-screw extruded at CARTIF (Valladolid, Spain) and the effects of extrusion were evaluated regarding bioactive properties. Commercial extruded rice was used as external control. Raw materials, non-extruded, and extruded samples were extracted with ethanol/water (80:20, v:v) and purified using C18 SepPak® Vac 3 cc cartridge [4,5], in order to obtain an extract rich in bioactive compounds. The cytotoxicity, anti-inflammatory, and antimicrobial activity were evaluated following previously reported procedures [6, 7]. Carob, commercial extruded rice and most of the extruded samples, showed cytotoxicity in the majority of the tumour cell lines tested (HeLa - cervical carcinoma, HepG2 - hepatocellular carcinoma, MCF-7 - breast adenocarcinoma, and NCI-H460 - non-small cell lung cancer). While, bean, rice, and most of the non-extruded samples presented no toxicity (GI50 value >400 µg/mL) using a non-tumour porcine liver cell culture (PLP2). In general, it was observed that extrusion process improves the cytotoxic potential in the rice-legumes sample mixtures, revealing lower GI50 concentrations (ranging between 115 and 362 µg/mL). The anti-inflammatory activity revealed a high heterogeneity, presenting bean and carob samples, the highest activity in comparison to the extruded samples. Concerning the antimicrobial activity, which was tested using a panel of multi-resistant isolated clinical strains, a low potential was observed, with non-extruded and extruded samples revealing higher values of minimum inhibitory concentration (MIC) and minimal bactericidal concentrations (MBC). In conclusion, rice-legumes flours, rice blends, beans, and carob fruits are a great alternative for the development of new gluten-free snacks products, in a market dominated mainly by cereals, due to the presence of different bioactive compounds, such as phenolic compounds, which can give healthier benefits to the consumers. The authors are grateful to FCT, Portugal and FEDER under Program PT2020 for financial support to CIMO (UID/AGR/00690/2013), L. Barros and R. Calhelha contracts. The authors are also grateful to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E, to the Spanish Ministry of Economy and Competitiveness (Project RTA2012-00042-C02) and INIA for the financial support of C. Arribas. info:eu-repo/semantics/publishedVersion

A preparação de infusões a partir de plantas aromáticas e medicinais é uma técnica utilizada desde a antiguidade [1]. As plantas podem ser utilizadas sozinhas ou misturadas para agregar valor nutricional, sabor ou potenciar alguma bioatividade [2,3]. Com este intuito, foram preparadas misturas de cinco espécies de plantas: Erica australis L. (EA), Genista tridentata L. (GT), Melissa officinalis, L. (MO), Mentha spicata L. (MS) e Prunella vulgaris L. (PV) em diferentes proporções (M1 – 20% PV, 40% GT e 40% MS; M2 – 30% EA, 30% PV e 40% MO; M3 – 30% PV, 35% MO e 35% MS). As misturas foram analisadas em termos de valor nutricional (açucares livres, HPLC-RI), composição fenólica (HPLC/DAD-ESI/MS) e bioatividade (propriedades antioxidantes, antimicrobianas, citotóxicas e anti-inflamatórias). A atividade antioxidante foi avaliada através da inibição da peroxidação lipídica (TBARS) e da hemólise oxidativa (OxHLIA); a atividade antimicrobiana foi testada em bactérias Gram + (Bacillus cereus e Listeria monocytogenes) e Gram - (Escherichia coli e Salmonela typhimurium) e fungos (Aspergillus niger, Aspergillus versicolor, Penicillium funiculosum e Penicillium verrucosum), usando o método da microdiluição em placa; para a avaliação da atividade citotóxica usou-se o método colorimétrico da sulforrodamina B em quatro linhas tumorais humanas (MCF-7, NCI-H460, HeLa e HepG2) e uma cultura primária de células não-tumorais de fígado de porco (PLP2); para a atividade anti-inflamatória, utilizou-se a linha de macrófagos RAW 246.7. A mistura M1 apresentou a maior quantidade de hidratos de carbono e de frutose bem como a maior concentração de compostos fenólicos totais, com prevalência de flavonóides, principalmente derivados de quercetina. Os resultados da atividade antioxidante e antibacteriana não foram significativos, no entanto a amostra M1 apresentou uma boa atividade antifúngica contra Penicillium funiculosum (MFC: 0,5 mg/mL), com um valor igual ao do controlo positivo de cetoconazol (MFC: 0,5 mg/mL). A M2 apresentou a melhor atividade citotóxica, em especial para a linha celular HepG2, com um GI50 de 166±10 μg/mL, e uma atividade anti-inflamatória moderada (GI50: 324±3 μg/mL). Este trabalho demonstrou o potencial das infusões de diversas misturas de plantas e o seu benefício para a saúde humana, bem como a sua possível aplicação na indústria alimentar ou farmacêutica. Os autores agradecem à Fundação para a Ciência e a Tecnologia (FCT, Portugal) e ao FEDER no âmbito do programa PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2019); financiamento nacional da FCT, PI, através do contrato de programa institucional de emprego científico para o contrato de L. Barros e R.C. Calhelha; o contrato de C. Pereira através da celebração contrato de programa previso no nº 4, 5 e 6 do artigo 23º do Decreto-Lei nº 57/2016, de 29 de agosto, alterado pela Lei nº 57/2017, de 19 de julho; FEDER-Interreg España-Portugal pelo apoio financeiro através do projeto 0377_Iberphenol_6_E; FEDER através do Programa Operacional Regional Norte 2020, no âmbito do Projeto NORTE-01-0145-FEDER-023289: DeCodE; e à empresa Ervital (Castro Daire, Portugal) pela cedência das amostras info:eu-repo/semantics/publishedVersion

Herbal teas, particularly infusions with both medicinal and nutritional purposes, have a long history of traditional uses among different cultures and regions [1]. The family Lamiaceae is an important medicinal plant family that is widely used in traditional medicine as a cure for various disorders [2]. In order to characterize and study the bioactivities of individual plant species of that family, three different plant species were selected: Melissa officinalis, L. (leaves), Mentha spicata L. (inflorescences), and Prunella vulgaris L. (leaves). Infusions were prepared from each plant species being further analyzed in terms of nutritional value, phenolic composition, and bioactive properties. The antioxidant activity was evaluated through two different in vitro assays: inhibition of lipid peroxidation in brain homogenates by thiobarbituric acid reactive substances (TBARS) and oxidative hemolysis inhibition assay (OxHLIA). The cytotoxic activity was assayed using MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung carcinoma), HeLa(cervical carcinoma) and HepG2 (hepatocellular carcinoma) human tumor cell lines, and also in a non-tumour cell line (porcine liver primary cells, PLP2). The phenolic profile was accessed through and UPLC-DAD-ESI/MS system. M. officinalis presented a high percentage of carbohydrate with sucrose being the predominant free sugar and no macronutrients were found in P. vulgaris infusion. Regarding phenolic compounds’ composition, the major detected compound in the three infusions was rosmarinic acid. Regarding, the bioactive properties, P. vulgaris and M. officinalis presented the best results for TBARS and OxHLIA assays, respectively. M. spicata showed the best anti-inflammatory activity and cytotoxic properties, with lowest GI50 values for all the tumour cell lines tested. None of the infusions revealed hepatotoxicity for non-tumour cells (PLP2). In general, M. spicata also presented the strongest antimicrobial activity. Thus, the complete characterization of these individual species confirms that they are excellent sources of bioactive molecules, especially phenolic acids. info:eu-repo/semantics/publishedVersion

A polpa dos frutos de Juçara (Euterpes edulis Martius), uma árvore nativa da mata Atlântica, é amplamente consumida graças ao seu sabor e valor nutricional, gerando grande quantidade de resíduos sólidos (casca) que geralmente são descartados. Este trabalho, teve por objetivo avaliar o perfil fenólico e bioatividades da casca de Juçara. Um total de dezenove compostos fenólicos foram identificados, sendo dezessete compostos fenólicos não-antocianinas (dois ácidos fenólicos, quatro flavanonóis, seis flavonas e cinco flavonóis). A casca do fruto de E. edulis apresentou atividades antioxidante e antimicrobiana e não apresentou hepatotoxicidade. Os resíduos da fruta Juçara podem ser utilizados para produzir aditivos alimentares de alto valor agregado, tanto corantes quanto conservantes, seguindo o conceito da bioeconomia circular. info:eu-repo/semantics/publishedVersion

Rosmarinus officinalis L. (Lamiaceae) is a plant native to the Mediterranean, popularly known as rosemary. In addition to its culinary use, it has also been used for therapeutic purposes since antiquity [1]. The European Union has considered rosemary extract as a safe and efficient food preservative, being rosmarinic acid (RA) its main constituent [2]. Extracts rich in RA not only have anti-inflammatory, antioxidant and hepatoprotective effects [1], but also prevent oxidation in foods without compromising their sensory acceptance. However, information on the stability of bioactive compounds in rosemary extract is scarce. The aim of this work was to investigate the potential phytochemical losses occurring throughout the sequential steps of in vitro gastrointestinal digestion and colonic fermentation of a rosemary aqueous extract. For this purpose, the crude (CE), digested (DE) and fermented (FE) extracts were characterized in terms of their phenolic profile and biological activities. Rosmarinic acid was the phytochemical that underwent the most significate transformation during digestion and fermentation, which amounted to 60% compared to the 26% degradation of the total phenolic compounds. Overall, the simulated digestion step decreased the antioxidant activity estimated by DPPH, ABTS, FRAP, ORAC and TBARS assays. Both CE and DE did not present anti-proliferative potential, however, FE exhibited a pronounced cytotoxic activity (GI50 = 116 μg/mL) against HeLa cervical carcinoma cell line. CE and DE showed to be moderate inhibitors of methicillin resistant Staphylococcus aureus (MRSA), methicillin-susceptible S. aureus (MSSA), and Listeria monocytogenes, whilst the FE acted as a moderate inhibitor of MRSA and MSSA. In view of the relevant biological activities confirmed in this study, it is possible to suggest the application of the rosemary aqueous extract as a food additive, either as a preservative and/or as a functional ingredient. However, the use of RA-rich formulations in food industry should consider the stability and bioavailability of this bioactive compound. info:eu-repo/semantics/publishedVersion

Tabebuia impetiginosa (Mart. ex DC.) Standl. has been used in traditional medicine for many centuries, being nowadays marketed as dried plant material (inner bark) for infusions, pills, and syrups. The main objective of the present work was to validate its popular use through the bioactivity evaluation of the inner bark (methanolic extract and infusion) and of two different formulations (pills and syrup) also based on the same plant-material. The antioxidant activity was evaluated by in vitro assays testing free radical scavenging activity, reducing power and inhibition of lipid peroxidation in brain homogenates. The cytotoxicity was determined in four human tumor cell lines (MCF-7, NCI-H460, HeLa and HepG2, and also in non-tumor cells (porcine liver primary cells, PLP2)). Furthermore, the sample was chemically characterized regarding free sugars, organic acids, fatty acids, and tocopherols. Syrup and methanolic extract showed the highest antioxidant activity, related to their highest amount of phenolics and flavonoids. Methanolic extract was the only sample showing cytotoxic effects on the tested human tumor cell lines, but none of the samples showed toxicity in PLP2. Glucose and oxalic acid were, respectively, the most abundant sugar and organic acid in the sample. Unsaturated predominated over the saturated fatty acids, due to oleic, linoleic, and linolenic acids expression. α- and γ-Tocopherols were also identified and quantified. Overall, T. impetiginosa might be used in different phytoformulations, taking advantage of its interesting bioactive properties and chemical composition., The authors are grateful to Foundation for Science and Technology (FCT, Portugal) for financial support to the research centre CIMO (strategic project PEst-OE/AGR/UI0690/2011), L. Barros researcher contract under “Programa Compromisso com Ciência–2008” and R.C. Calhelha grant (SFRH/BPD/BPD/68344/2010)., info:eu-repo/semantics/publishedVersion

Several novel methyl 7-[(hetero)arylamino]thieno[2,3-b]pyrazine-6-carboxylates were synthesized by Pd-catalyzed C–N Buchwald–Hartwig cross-coupling of either methyl 7-aminothieno[3,2-b]pyrazine-6-carboxylate with (hetero)arylhalides or 7-bromothieno[2,3-b]pyrazine-6-carboxylate with (hetero)arylamines in good-to-excellent yields (50% quantitative yield), using different reaction conditions, namely ligands and solvents, due to the different electronic character of the substrates. The antitumoral potential of these compounds was evaluated in four human tumor cell lines: gastric adenocarcinoma (AGS), colorectal adenocarcinoma (CaCo-2), breast carcinoma (MCF7), and non-small-cell lung carcinoma (NCI-H460) using the SRB assay, and it was possible to establish some structure–activity relationships. Furthermore, they did not show relevant toxicity against a non-tumor cell line culture from the African green monkey kidney (Vero). The most promising compounds (GI50 ≤ 11 µM), showed some selectivity either against AGS or CaCo-2 cell lines without toxicity at their GI50 values. The effects of the methoxylated compounds 2b (2-OMeC6H4), 2f and 2g (3,4- or 3,5-diOMeC6H3, respectively) on the cell cycle profile and induction of apoptosis were further studied in the AGS cell line. Nevertheless, even for the most active (GI50 = 7.8 µM) and selective compound (2g) against this cell line, it was observed that a huge number of dead cells gave rise to an atypical distribution on the cell cycle profile and that these cells were not apoptotic, which points to a different mechanism of action for the AGS cell growth inhibition., This research was funded by Fundação para a Ciência e Tecnologia (FCT)—Portugal, which financially supports CQUM (UID/QUI/686/2019), and also financed by the European Regional Develop- ment Fund (ERDF), COMPETE2020 and Portugal2020, and the PTNMR network also supported by Portugal2020. J.M.R. PhD grant (SFRH/BD/115844/2016) was financed by FCT, ESF (European Social Fund—North Portugal Regional Operational Program) and HCOP (Human Capital Operational Program). The authors are grateful to FCT, Portugal, for financial support through national funds FCT/MCTES to the CIMO (UIDB/00690/2020). L.B. and R.C.C. thank the national funding by FCT, Portugal, through the institutional scientific employment program-contract for their contracts.

In the present work, sage (Salvia officinalis L.) and basil (Ocimum basilicum L.) were exploited for their preservative purposes, as viable alternatives to artificial ones. The ultrasound-assisted extraction (UAE) of bioactive compounds was pre-optimized using factorial screening analysis, prior to applying response surface methodology (RSM). The obtained extracts were characterized in terms of phenolic compounds by high-performance liquid chromatography coupled to photodiode array detector and mass spectrometer HPLC-DAD-ESI/MS and bioactivities, namely the antioxidant, antimicrobial and cytotoxic potential. In addition, the most promising extracts were incorporated into yogurts, that were further screened for nutritional and physico-chemical properties and microbial load, over a shelf life of 14 days. According to the obtained results, the solvent percentage is the most relevant factor for obtaining rosmarinic acid-rich extract, followed by the extraction time and ultrasonic power. For the antioxidant and antimicrobial activity, sage showed the best result for both analysis and none of the two plant extracts were hepatotoxic. Finally, both extracts did not show changes in the physicochemical and nutritional characteristics of the yogurts and did not interfere with the growth of lactic acid bacteria, an important microorganism during yogurt fermentation. These results highlight the high potential of sage and basil as natural preservatives. Please add: This research was funded by the Portuguese Foundation for Science and Technology (FCT) and ERDF, through the Incentive System to Research and Technological development, within the Portugal2020 Competitiveness and Internationalization Operational Program for the financial support to BIOMA: POCI_01_0247_FEDER_046112; ERDF through the Regional Operational Program North 2020, within the scope of Project GreenHealth: Norte-01-0145-FEDER-000042. FCT: Portugal for financial support through national funds FCT/MCTES to the CIMO (UIDB/00690/2020) and to F.A. Fernandes PhD grant (SFRH/BD/145467/2019). L. Barros, M.I. Dias and R.C. Calhelha also thank the national funding by FCT, P.I., through the institutional scientific employment program-contract for their contracts and M. Carocho (CEECIND/00831/2018), J. Pinela (CEECIND/01011/2018) and S.A. Heleno (CEECIND/03040/2017) to the national funding by FCT, P.I., through the individual scientific employment program-contracts; ERDF, through the Incentive System to Research and Technological development, within the Portugal2020 Competitiveness and Internationalization Operational Program for the financial support to BIOMA: POCI_01_0247_FEDER_046112; ERDF through the Regional Operational Program North 2020, within the scope of Project GreenHealth: Norte-01-0145-FEDER-000042. Serbian Ministry of Education, Science and Technological Development [Contract No. 451-03-68/2020-14/200007] for MI and MS. info:eu-repo/semantics/publishedVersion

A produção de vinho representa hoje em dia uma das maiores atividades agroindustriais a nível mundial. Como consequência gera uma enorme quantidade de subprodutos (bagaço e borras) que representam por vezes até 30% das uvas vinificadas, sendo a maioria completamente descartados sem tratamento adequado e/ou outras finalidades [1-4]. O bagaço de uva representa, por isso, um abundante e acessível subproduto industrial que contém uma grande variedade de compostos bioativos, nomeadamente fenólicos, que têm sido relacionados com benefícios para a saúde do consumidor [l]. O objetivo deste estudo foi mimetizar a digestão gastrointestinal e a fermentação colónica de bagaço de uva de Merlot, de forma a avaliar uma possível redução do conteúdo em fitoquímicos, correlacionando o perfil de compostos fenólicos com as suas atividades biológicas. Assim, foram caracterizados três extratos de bagaço (inicial, digerido e fermentado) relativamente ao seu conteúdo em compostos fenólicos antociânicos e não antociânicos por HPLC-DAD-ESI/MS. Além disso, foi também a valiado o seu potencial antioxidante, antibacteriano e citotóxico para células tumorais e não-tumorais. Os compostos fenólicos mais abundantes identificados nas três amostras estudadas foram um dímero de (epi)catequina tipo B, (+)-catequina e (-)-epicatequina. Foram identificados vinte compostos fenólicos não antociânicos na amostra inicial (66 mg/g de extrato), tendo sido significativamente reduzidos para 11 compostos após digestão invitro. Foram identificadas cinco antocianinas, no entanto, as quantidades diminuíram imenso após digestão e fermentação. Pelos resultados obtidos podemos concluir que o processo de digestão in vitro promoveu drásticas reduções qualitativas e quantitativas no perfil de compostos fenólicos no extrato micial de bagaço de uvas Merlot. Tais alterações podem estar relacionadas com a diminuição de algumas bioatividades do extrato, como é o caso das propriedades antioxidantes e antibacterianas, embora não de forma diretamente proporcional. No entanto, a fermentação colónica teve um efeito positivo sobre o potencial citoóxico do extrato em linhas celulares tumorais humanas. Há ainda um conhecimento muito escasso sobre a estabilidade dos compostos fenólicos de Bagaço de uva e propriedades bioativas durante a digestão gastrointestinal e fermentação colónica, no entanto, os resultados obtidos poderão ser de grande utilidade para o desenvolvimento de novos suplementos nutracêuticos e produtos alimentares funcionalizados. FCT e FEDER sob o programa PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2013) e contratos de L. Barros e R. Calhelha. FEDER-Interreg Espana-Portugal (0377 Iberphenol_6_E). V.G. Corrêa ao CAPES. R.M. Peralta (307944/2015-8) e A. Bracht(304090/20Ï6-6). info:eu-repo/semantics/publishedVersion

O cozimento por extrusão é um tipo de processamento alimentar de curta duração que se caracteriza pelo uso de alta temperatura. Prima também pelo controle automatizado, alta capacidade de produção, operação contínua, alta produtividade, versatilidade e alta adaptabilidade às condições de processamento, eficiência energética e baixo custo [ 1,2]. Esta tecnologia é útil para obter produtos tais como, salgadinhos, cereais matinais prontos para consumo, biscoitos, amidos modificados, proteínas vegetais texturizadas, massas, substitutos de carne e alimentos para animais de estimação [3]. Neste estudo, foram desenvolvidas seis misturas com diferentes proporções de arroz, feijão e alfarroba, avaliando-se as alterações induzidas pelo cozimento por extrusão na sua composição fenólica. Paraa valiaçãod op erfilf enólico,a sa mostrasf orams ubmetidasa extraçõesh idroetanólicas (80:20, vlv) que foram, posteriormente, analisadas por HPLC-DAD-ESI/MS. O perfil fenólico revelou a ausência de compostos nas amostras de arroz e arroz comercial, contudo, evidenciou a presença de 5 e 28 compostos nas amostras de feijão e alfarroba, respetivamente. Na amostra de feijão destacou-se o derivado do ácido p-cumánico com um valor de 121 g/100 g massa seca (ms), ao contrário da amostra de alfarroba onde o composto maioritário foi um tetragaloil-glucósido, com 763 g/100 g ms. Relativamente às misturas, verificou-se que a percentagem de leguminosas nas formulações foi uma variável que afetou significativamente a concentração de compostos fenólicos, uma vez que foi encontrada maior quantidade desses compostos nas misturas com maior concentração de feijão e alfarroba. Tendo em conta os efeitos da tecnologia em estudo, a análise estatística mostrou uma diferença significativa entre todas as amostras, extrusadas e não-extrusadas, com exceção das formulações com 40% de feijão e 60% de arroz, onde o processo de extrusão não causou mudanças relevantes no conteúdo de compostos fenólicos. Deste modo, foi evidente a presença de compostos bioativos nas farinhas de arrozleguminosas (misturas de arroz, feijão e alfarroba), traduzindo-se numa ótima alternativa para o desenvolvimento de novos lanches sem glúten, fazendo frente a um mercado dominado principalmente por cereais. Os autores agradecem à Fundação para a Ciência e a Tecnologia (FCT) e ao FEDER no âmbito do programa PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2013), pelo contrato de L. Barras. Os autores também agradecem ao programa FEDER-Interreg Espana-Portugal pelo apoio financeiro através do projeto 0377_Iberphenol_6_E e ao Ministério Espanhol da Economia e Competitividade (Projeto RTA2012-00042-C02) e INIA pelo financiamento de C. Arribas. info:eu-repo/semantics/publishedVersion

For several centuries, wild mushrooms have been part of the normal human diet, and also extensively consumed due to their organoleptic, chemical and nutritional properties. Not just fruiting bodies, but also their mycelia have been exploited for the development of natural drugs. Pleurotus eryngii (DC.) Quél and Suillus bellinii (Inzenga) Watling, in particular, are interesting source of nutritional and bioactive compounds and properties. In this context, the present work deals with the production of mushrooms’ mycelium using different solid and liquid culture media (PDA, PDB and solid and liquid iMMN). Furthermore, phenolic acids and ergosterol were determined both in fruiting bodies and mycelia, by high performance liquid chromatography coupled to a photodiode array detector (HPLC-PAD) or an ultraviolet detector (HPLC-UV), respectively, and the in vitro anti-inflammatory effects were evaluated by measuring the down-regulation of NO production in LPS-stimulated RAW264 cells[1]. P. eryngii mycelia, under specific culture conditions, showed higher anti-inflammatory activity than the corresponding fruiting body. Otherwise, S. bellinii fruiting body had a strong anti-inflammatory activity which might be related to its higher contents in phenolic acids despite the higher amounts of ergosterol observed in its mycelium. Globally, the obtained mycelia maintained the functional components of the parent mushroom, namely phenolic acids and ergosterol, as also the anti-inflammatory properties, and could be used in nutraceutical or pharmaceutical formulations. N/A

The consumers' interest for a healthier diet and the use of natural products in the prevention and/or treatment of health problems has triggered an intensive search for bioactive plant compounds. Plants are a good source of phenolic compounds, terpenoids, alkaloids, quinones, carotenoids, sterols, glucosinolates and other compounds, which provide health benefits due to their wide range of biological properties. This work aimed to evaluate the effects of gamma radiation on the phenolic composition, cytotoxicity, virucidal and antimicrobial properties of Aloysia citrodora L. and Mentha x piperita L. infusions. Gamma radiation treatment was performed in a 60Co chamber, applying the doses of 1 and 10 kGy and the results were compared with the control sample (0 kGy). The phenolic compounds profile was obtained by HPLC-DAD-ESI/MS and the cytotoxicity was evaluated in human tumour cell lines and in a primary cell culture. For virucidal efficacy evaluation, MNV-1 and HAdV-5 viral titers were determined by plaque assay in Raw264.7 and A549 cell lines, respectively. The antibacterial activity was evaluated against Gram-negative and Gram-positive bacteria. Radiation treatment has been shown to cause different effects on phenolic concentrations and virucidal efficacy, depending on the plant species studied, the virus and the absorbed dose. In cytotoxicity assay A. citrodora did not reveal significant statistically differences considering the different doses applied, however M. piperita at 10 kGy stood out with the highest cytotoxic potential in all tested cell lines. Regarding antibacterial assays, S. aureus was the only bacteria that was sensible to the infusions of non-irradiated and 1 kGy irradiated in both species. Thus, this study contribute to a wider knowledge of the effects of gamma irradiation on several relevant bioactive potentials of some aromatic and medicinal plants and showed that gamma radiation is a technique recommended for these matrices. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013) and CTN (RECI/AAG-TEC/0400/2012). The authors are also grateful to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion

The concept of bio-economy has emerged to overcome some sustainability challenges, and this involves conversion of agricultural residues and waste streams into high value-added products that can be utilized as ingredients for several bio-based industrial processes, delivering both economic growth, scientific interest and a better environment. The present work was carried out to re-utilize discarded Agaricus blazei Murill. Its nutritional composition was evaluated and ethanolic extracts were prepared by Soxhlet extraction to be further evaluated for their anti-inflammatory, anti-tyrosinase and cytotoxic properties against different tumor cell lines (SRB assay). MTT and LDH assays were also used to determine cell viability and cell death respectively in Caco-2 and HT29 cells lines. Essential nutrients such as carbohydrates, proteins and fat, were found; twenty fatty acids were detected, sugars (mannitol and trehalose), α-tocopherol and oxalic acid were also present. The extracts, up to 100 μg/mL, were able to maintain viability of Caco-2 and HT29 cells. The extracts also presented anti-tyrosinase activity (EC50 1.33±0.02 mg/mL) and with no toxicity in tumor cells. The results obtained suggested that the extracts obtained from Agaricus blazei Murill residues can be utilized as an inexpensive and sustainable source of nutraceutical and functional food ingredients. FCT for financial support to CIMO (UID/AGR/00690/2013); European Agricultural Fund for Rural Development (EAFRD), through the Rural Development Program (PDR2020), within the scope of Project MicoCoating (PDR2020-101-031472). This work also received financial support from the European Union (FEDER funds through COMPETE), under the Partnership Agreement PT2020, and National Funds (FCT, Foundation for Science and Technology) through project LAQV/UID/QUI/50006/2013 and NORTE-07-0124-FEDER-000069 – Food Science. info:eu-repo/semantics/publishedVersion

Myrtus nivelli ileaves are widely used in the Algerian folk medicine, due toits healthpromoting properties attributed to the phenolic composition. In this context, bioactivities of the aqueous extract and its organic sub-fractions (ethyl acetate and butanol) were evaluated with focus onantioxidant properties (free radicals’ scavenging activity, reducing power, inhibition of β-carotene blanching andlipid peroxidation), anti-inflammatory potential. (inhibition of NO production in lipopolysaccharide-stimulated RAW 264.7 macrophages), cytotoxicity for human tumor cells and normal porcine liver primary cells, and antibacterial activity against ten clinical isolates. The obtained results indicated high activities for all the tested samples. The antioxidant activity was higher than that obtained for Trolox (positive control), showing the ethylacetate fraction the highest activity as also the highest inhibition of NO production (EC50= 104 ± 6 μg/mL) and cytotoxicity for all the tested tumor cell lines (HeLa- 15.39±1 μg/mL, MCF-7- 16.12± 1μg/mL, NCI-H460-, HepG2-17.67±1 μg/mL and 49.01 ± 3 μg/mL). The butanol fraction and the crude aqueous extract presented the highest antibacterial activity. Overall, this study highlights the potential of M. nivellii leaves in the preparation of bioactive ingredients. info:eu-repo/semantics/publishedVersion

O interesse em espécies silvestres tem vindo a aumentar gradualmente devido à sua rica composição nutricional e moléculas de interesse bioativo, fundamentais para o bom funcionamento do organismo. Esta diversidade de compostos, para além de ser benéfica a nível alimentar, estabelece vantagens para futuras aplicações em diversos setores industriais, particularmente, alimentar, cosmético e farmacêutico [1,2]. A espécie Rubus ulmifolius Schott (vulgarmente denominada por silva-brava ou amora-silvestre) pertence à família Rosaceae e é essencialmente conhecida pêlos seus frutos. O seu perfil químico evidencia compostos bioativos, nomeadamente, compostos fenólicos, que lhes confere propriedades antioxidantes, anti-inflamatórias, antimicrobianas, antitumorais, entre outras [3, 4]. Neste trabalho, foi determinado o perfil fenólico dos frutos da espécie R. ulmifolius, assim como, avaliado o seu potencial como agente bioativo, estudando a atividade citotóxica e antimicrobiana. Os compostos fenólicos foram determinados através de um sistema HPLC-DAD-ESI/ MS, enquanto que o potencial citotóxico das amostras foi avaliado em linhas celulares tumorais MCF-7 (adenocarcinoma da mama), NCI-H460 (carcinoma de pulmão), HeLa (carcinoma de cervical) e HepG2 (carcinoma hepatocelular), e numa cultura de células primárias não-tumorais (PLP2), pelo método da sulforrodamina B (SRB). A atividade antimicrobiana foi avaliada através do método de microdiluição usando bactérias Gram-positivo e Gram-negativo, bem como fungos. Tendo em conta os resultados obtidos, R. ulmifolius evidenciou um perfil fenólico com 11 compostos não antociânicos (salientando-se o pentósido do ácido elágico) e 5 compostos antociânicos (destacando-se a cianidina-3-O-glucósido) com potencial corante. No que concerne às suas propriedades bioativas, os extratos hidroetanólicos preparados a partir dos frutos não revelaram qualquer capacidade antiproliferativa em nenhuma das linhas celulares testadas, mas também não manifestaram toxicidade nas células não-tumorais (GI50>400). No entanto, quanto à atividade antimicrobiana obtiveram-se resultados favoráveis, tendo os extratos revelado um efeito bacteriostático, com valores de MIC (concentração mínima inibitória) entre 5 e 20 mg/mL. Assim, este estudo mostrou que os frutos de R. ulmifolius são uma boa fonte de compostos fenólicos e que exibem propriedades antimicrobianas, sendo possível a sua aplicabilidade não só como fonte de nutrientes, mas também como agentes bioativos. Fundação para a Ciência e a Tecnologia (FCT, Portugal) e FEDER no âmbito do programa PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2013) e contratos de L. Barras e R. Calhelha. Programa FEDER-Interreg Espana-PortugaI pelo apoio financeiro através do projeto 0377_Iberphenol_6_E. Este trabalho foi financiado pelo Fundo Europeu de Desenvolvimento Regional (FEDER) através do Programa Operacional Regional Norte 2020, no âmbito do Projeto NORTE-01-0145- FEDER-023289( DeCodE) e projeto Mobilizador Norte-01-0247-FEDER-024479: ValorNatural. info:eu-repo/semantics/publishedVersion

Currently, the food industry is interested in replacing artificial additives by natural ingredients. Some plant extracts have emerged as possible alternatives to artificial preservatives, namely antioxidants. In fact, dairy, meat and bakery products have been developed, incorporating extracts of aromatic plants, spices or fruits, which have antioxidant properties. In this work, the preserving effectiveness of an extract rich in rosmarinic acid was tested in cupcakes and compared to an artificial additive (potassium sorbate, E202). The extract was obtained from Melissa officinalis L. (lemon balm) by applying an ultrasound technique using a mixture of ethanol/water as the extraction solvent. After confirming its antioxidant properties (free radical scavenging effect EC50 = 79 ± 2 μg/mL; reducing power EC50 = 49 ± 1 μg/mL), antimicrobial (against 8 bacteria and 8 food contaminating fungi), and absence of toxicity (in cell lines), it was incorporated in cupcakes, and analysed immediately after incorporation and after 3 and 5 days of storage at room temperature and protected from light. All samples were analysed chromatographically in terms of fatty acids (GC-FID) and free sugars (HPLC-RI). Regarding fatty acids, a total of 21 molecules were identified, with predominance of saturated fatty acids in all cupcakes samples. Individually, palmitic acid and oleic acid were detected in the highest percentages. Among free sugars, sucrose (the major form) and glucose were identified in all samples. The results demonstrate that the addition of the extract rich in rosmarinic acid caused no changes in fatty acids and sugars’ profiles, having the potential to be used in pastry products, meeting the current consumers demand. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013), REQUIMTE (UID/QUI/50006/2013 - POCI/01/0145/ FERDER/007265) and C. Caleja (SFRH/BD/93007/2013) grant. This work is funded by the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project NORTE-01-0145-FEDER-023289: DeCodE. info:eu-repo/semantics/publishedVersion

Na indústria alimentar, a substituição de aditivos artificiais por compostos naturais é uma tendência muito atual [1]. Além dos efeitos específicos, os ingredientes obtidos naturalmente, apresentam frequentemente importantes níveis de bioatividade [2]. Neste trabalho, pretendemos demonstrar o potencial de utilização do extrato etanólico de flores de Castanea sativa Mill. como conservante natural. Trigaloil-HHDP-glucósido e quercetina-3-O-glucurónido foram o elagitanino e o flavonóide maioritário, respetivamente, num total de 14 compostos identificados por HPLC-DAD-ESI/MS. O extrato demonstrou atividade antioxidante e antimicrobiana e, após incorporação em pastéis de natas, o seu efeito foi comparado com o sorbato de potássio (E202, conservante artificial). Os resultados foram comparados em dois períodos: no dia de preparação e após dois dias de armazenamento. Os resultados obtidos demonstram que as flores de castanheiro podem ser utilizadas em produtos de pastelaria, substituindo conservantes artificiais e melhorando as propriedades funcionais do produto sem, no entanto, provocar alterações nos perfis nutricionais e físico-químicos conduzindo, no entanto, a alterações na sua aparência. Os autores agradecem à Fundação para a Ciência e a Tecnologia (FCT, Portugal) e ao FEDER no âmbito do programa PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2013), REQUIMTE (UID/QUI/50006/2013 - POCI/01/0145/ FERDER/007265), bolsa de C. Caleja (SFRH/BD/93007/2013) e contratos de L. Barros e J.C.M. Barreira. Este trabalho é financiado pelo Fundo Europeu de Desenvolvimento Regional (FEDER) através do Programa Operacional Regional Norte 2020, no âmbito do Projeto NORTE-01-0145-FEDER-023289: DeCodE .Os autores também agradecem ao programa FEDER-Interreg España-Portugal pelo apoio financeiro através do projeto 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion

Atualmente, uma das principais causas de morte são as doenças de foro oncológico. As terapias convencionais (cirurgia, radioterapia e quimioterapia) têm como inconveniente a falta de seletividade que resulta, por norma, em efeitos secundários diversos. Assim, existe a necessidade de explorar terapêuticas alternativas, surgindo desta forma a terapia fotodinâmica (PDT, do mglês, Photodynamic Jherapy) [l]. Esta tem por base a fotocitotoxicidade de diferentes fotossensibüizadores (FS) que se acumulam, preferencialmente e de forma seletiva nas células tumorais, e que surge apenas após excitação com luz de comprimento de onda adequado [2]. A PDT tem como base processos fotodmâmicos, em que a luz absorvida pelo fotossensibilizador gera um estado eletrónico excitado que, por sua vez, pode transferir a sua energia para o oxigénio no estado fundamental, produzmdo assim oxigénio smguleto citotóxico. Este, dá origem a imia cascata de processos bioquímicos conducentes à morte das células tumorais [3]. Uma dasse de compostos que tem suscitado bastante interesse como FS são os corantes esquarflicos devido às propriedades específicas que apresentam [4]. Neste ü-abalho, foram sintetizados alguns corantes aminoesquarilicos catiónicos derivados do 2-metilbenzotiazole e da 2-metiquinolina, adequadamente substituídos no anel central com uma molécula de colesterol (figurai), como esti-atégia para potenciaknente melhorar a permeabilidadec elular,e feitaa respetivac aracterizaçãoe stmtairaLA (foto)citotoxicidaded os corantes sintetizados foi testada in vitro em várias linhas celulares hunorais humanas (carcmomas da mama, puhnão, cervical e fígado) e comparada com a dos corantes isentos de colesterol. A citotoxiddade dos corantes também foi avaliada numa linha de células primárias não tumorais de fígado de porco, na presença e ausência de luz. Foi iguaknente avaliada a capacidade de produção de oxigénio singuleto dos corantes sintetizados. Assim, neste trabalho pretendeu-se desenvolver novos compostos com absorção no W próximo e boa penneabilidade celular que possams erp otenciaisf otossensibilizadoresp arat erapiaf otodmâmica. Os resultados obtidos mosü-am que os corantes smtetizados não apresentam praticamente citotoxicidade no escuro, nas concenü-ações usadas; contudo, sob radiação os corantes que possuem o elemento de reconhecknento celular não parecem mostrar desempenho significativamente melhor que o dos mesmos corantes sem o colesterol. info:eu-repo/semantics/publishedVersion

The Amazon rainforest is the home to several medicinal plant species popularly used for the treatment of a wide range of diseases. Despite the rich biodiversity of this region, there is still a lack of comprehensive studies regarding the uses, bioactivities, chemical composition and pharmacological properties of many of these medicinal plants [l]. In the present work, Garcinia brasiliensis Mart., a green plant native to the Amazon region, cultivated for its edible fruit, popularly referred to as "bacuparianão" or "guapomo" was studied. Some of its well reported bioactive properties include antioxidant, antidiabetic, anti-inflammatory, antispasmodic and antiprotozoal activities but very little studies have been conducted describing the phytochemical profile and establishing compounds-bioactivity relationships of the extracts obtained from its vegetative organs [2]. In view of these pharmaceutical potentials, leaves of G. brasilienses were collected at São Cristóvão site, Paraná, Brazil. They were then submitted to a cold methanol extraction, and further liquid-liquid partitioned to obtain hexane, dichloromethane and ethyl acetate fractions. The antioxidant, antiinflammatory, antimicrobial and cytotoxic activities of each fraction were tested, and the most bioactive fraction was submitted to phenolic characterization using HPLCDAD- ESI/MSn. The results showed that the ethyl acetate fraction showed the highest antioxidant activity which was comparable to Trolox, the positive control. This fraction also showed a broad spectrum antimicrobial activity against Gram-positive and Gramnegative bactéria, and Cândida albicans. The phenolic profile of the ethyl acetate fraction presented twelve flavonoids, being morelloflavone-7"-0-glucoside (52. 1±0.4 mg/g) and gardinia biflavonoid 2a glucoside (27. 5±0.2mg/g) the major compounds identified. These results indicate that leaves of G. brasiliensis might be a potential source of natural biomolecules for pharmaceutical and medicinal applications. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financiai support to CIMO (UID/AGR/00690/2013) and L. Barros contract; to FEDER-Interreg Espana-Portugal programme for financiai support through the project 0377_Iberphenol_6_E.] info:eu-repo/semantics/publishedVersion

This study was carried out to evaluate the antibacterial, antifungal and cytotoxic effects of chloroform, ethyl acetate and aqueous fractions obtained from a methanol 80 % extract of Cestrum nocturnum L. (family Solanaceae) aerial parts, and to investigate the bioactive constituents present in these fractions. The results showed that the chloroform fraction had the highest antibacterial effect and cytotoxicity against lung, colon and hepatocellular carcinoma cell lines, while the ethyl acetate fraction displayed the highest antifungal activity and cytotoxicity against cervical carcinoma cell lines. Phytochemical analysis showed the presence of triterpenes, coumarins and flavonoids in the chloroform fraction. Flavonoids, triterpenes and carbohydrates in the ethyl acetate fraction while tannins, flavonoids, saponins, carbohydrates and triterpenes were detected in the aqueous fraction. β-sitosterol, stigmasterol, protocatechuic acid and apigenin were isolated from the chloroform fraction; Kaempferol 8-O-methyl ether, kaempferol, kaempferol 3-O-α-rhamnoside and luteolin 7-O-β-glucoside were isolated from the ethyl acetate fraction, while kaempferol di-sugars: kaempferol 3-O-β-glucoside-7-O-α-rhamnoside and kaempferol 3,7-di-O-α-rhamnoside, were isolated from the aqueous fraction. This study proves the bioactivities of C. nocturnum aerial parts that can be explored as a source of medicinal compounds. The authors are grateful to the strategic project PEst-OE/AGR/UI0690/2011 for the financial support to CIMO. R. Calhelha thanks to FCT, POPH-QREN and FSE for his grant (SFRH/BD/78307/2011) and the Serbian Ministry of Education and Science, grant number 173032 for financial support. info:eu-repo/semantics/publishedVersion

Geranium molle L. (Fig. l) has been used in the Portuguese folk medicine for the treatment of various ailments including cancer [l]. However, contrary to many Geranium species, the bioactivity and the phytochemical composition of G. molle are virtually unexplored. The first phase of this study consisted in the chemical characterization (nutritional value, free sugars, organic acids, fatty acids and tocopherols) of a G. molle sample from Trás-os-Montes, north-eastern Portugal. The bioactive properties of infusion and decoction (common forms of consumption) and of different organic extracts obtained by sequential extraction were also evaluated. G. molle was shown to be rich in carbohydrates and proteins, providing tocopherols and essential fatty acids. Amongst the various extracts, the acetone extract was found to have the highest content of phenolic compounds as well as the highest antioxidant and cytotoxic activities [2]. Following this preliminary evaluation, the most active extracts (acetone and methanol) were fractionated by column chromatography and the resulting fractions were evaluated for their antioxidant and cytotoxic activities. The bio-guided fractionation of the extracts resulted in several fractions with improved bioactivity in comparison with the corresponding crude extracts. The fractions obtained from the acetone extract consistently displayed the lowest EC50 and GI50 values and presented the highest content of total phenolic compounds. The phytochemical composition of the most bioactive fractions of the acetone and methanol extracts was included about thirty identified compounds, mainly flavonoids and phenolic acids, which could be identified for the first time in G. molle [3]. Work supported under the projects POCI-01-0145-FEDER-006958, UID/AGR/00690/2013 (CIMO) and UID/QUI/00616/ 2013 (CQ-VR). V.C. Graça thanks FCT for her grant (SFRH/BD/52542/2014) and L. Barras contract. The authors are also grateful to Interreg Espana-Portugal for financiai support through the project 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion

Rosmarinus officinalis L., alecrim, pertence à família das Lamiaceae e encontra-se amplamente disseminado em vários países da região do Mediterrâneo [1]. É usado sobretudo como condimento, mas os seus extratos aquosos ricos em compostos fenólicos, maioritariamente ácido rosmarínico, têm um grande potencial para ser usados como conservantes e/ou ingredientes funcionais [2]. É, no entanto, necessário perceber a estabilidade e biodisponibilidade destes extratos aquosos e dos seus compostos bioativos após a digestão gastrointestinal e processos de fermentação colónica, de modo a não perder a funcionalidade dos mesmos. No presente trabalho, o extrato inicial e os extratos digeridos e fermentados de alecrim foram caracterizados em termos de compostos fenólicos por HPLC-DAD/ESI-MS e, posteriormente, estudados os seus potenciais antioxidante, antibacteriano e antiproliferativo. Foram identificados 16 compostos fenólicos, entre eles 10 ácidos fenólicos e 6 flavonoides, sendo o ácido rosmarínico o composto maioritário, como era expectável. Foi também este composto que mais sofreu os efeitos da digestão e fermentação representando um total de 60%, em comparação com os 26% dos compostos fenólicos totais. No geral, a digestão gastrointestinal in vitro diminuiu a atividade antioxidante obtida pelos métodos de DPPH, ABTS, FRAP, ORAC e TBARS. Tanto o extrato inicial como o digerido não apresentaram qualquer atividade antiproliferativa, no entanto, o extrato fermentado exibiu um ótima atividade antiproliferativa na linha celular HeLa (carcinoma cervical, GI50 = 116 μg/mL). Relativamente à atividade antibacteriana, o extrato inicial e o extrato digerido revelaram-se moderadamente eficazes contra o crescimento de Staphylococcus aureus, S. aureus resistente à meticilina (MRSA) e sensível à meticilina (MSSA) e também contra a Listeria monocytogenes. O extrato fermentado revelou-se também moderadamente eficaz contra o crescimento de MRSA e MSSA. Os resultados aqui obtidos revelam a importância dos extratos ricos em ácido rosmarínico como potenciais conservantes e ingredientes funcionais, no entanto, o uso de técnicas de estabilização/microencapsulação seria importante para assegurar uma adequada ação destes compostos após digestão e fermentação. FCT e FEDER sob o programa PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2013) e contratos de L. Barros e R. Calhelha. FEDER-Interreg España-Portugal (0377_Iberphenol_6_E). V.G. Correa ao CAPES. R.M. Peralta (307944/2015-8) e A. Bracht (304090/2016-6). info:eu-repo/semantics/publishedVersion

As plantas possuem na sua composição vários compostos de interesse, particularmente compostos fenólicos, que se classificam como moléculas bioativas, exercendo funções benéficas para a saúde humana [ l ]. Ocimum basilicum var. purpurascens (popularmente conhecido por manjericão vermelho rubi) é uma planta anual, pertencente à família das Lamiaceae, resultando a sua cor púrpura da elevada concentração em compostos antociânicos. Esta variedade de O. basilicum é usada como planta ornamental, embora o seu óleo essencial seja utilizado na medicina popular [2]. O objetivo deste trabalho consistiu em avaliar o conteúdo em compostos fenólicos, bem como as propriedades bioativas (atividade antioxidante, citotoxiddade e atividade antimicrobiana) de um extrato hidroetanólico (80:20, vi v) de manjericão vermelho rubi. A determinação do perfil fenólico foi realizada através de um sistema de HPLC-DAD-ESI/ MS, a atividade antioxidante foi avaliada através de três ensaios in vitro (atividade captadora de radicais DPPH, poder redutor e mibição da descoloração do (3-caroteno), a citotoxicidade foi determinada com recurso a linhas celulares tumorais (HepG2, NCI-H460, MCF-7 e HeLa), e a uma cultura de células primárias não tumorais (PLP2), aplicando o ensaio da sulforrodamina B e, por fim, a atividade antimicrobiana foi analisada através do método de microdiluição em bactérias Gram-positivo e Gram-negativo, e em fungos. No extrato de manjericão vermelho rubi foi evidente a presença de 26 compostos fenólicos, dos quais 13 foram identificados como compostos fenólicos não antociânicos, destacando-se o ácido rosmarínico como composto maioritário, e 13 compostos antociânicos, no qual a cianidina-3-(6, 6'-di-p-cumaroil)-soforosil-5-glucósido foi o composto mais abundante. Relativamente ao potencial bioativo desta espécie, o extrato revelou atividade antioxidante, evidenciando-se o ensaio da atividade captadora de radicais DPPH com valores de EC^ = 60 |^g/mL. Foi também notório, no ensaio da atividade citotóxica, a ausência de capacidade antiproliferativa em todas as linhas celulares testadas mas, também, ausência de toxicidade do extrato para células não tumorais (GI >400 |^g/mL). Relativamente à atividade antimicrobiana, os resultados obtidos foram satisfatórios para a maioria das bactérias testadas, mas também nos fungos; apresentando estes últimos valores de MIC (concentração mínima inibitória) inferiores, comparativamente às bactérias estudadas. Os resultados obtidos neste estudo permitem concluir que o manjericão vermelho rubi é uma planta rica em moléculas bioativas, nomeadamente compostos antociânicos e não-antociânicos, que suscitam elevado interesse ao nível de vários setores industriais, tal como o farmacêutico e o alimentar. Os autores agradecem à Fundação para a Ciência e a Tecnologia (FCT, Portugal) e ao FEDER no âmbito do programa PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2013), contratos de L .Barros e R. Calhelha.Os autores também agradecem ao programa FEDER-Interreg Espana- Portugal pelo apoio financeiro através do projeto 0377 Iberphenol_6_E. Este trabalho foi financiado pelo Fundo Europeu de Desenvolvimento Regional (FEDER) através do "Programa Operacional Regional 2020, no âmbito do Projeto NORTE-01-0145-FEDER-02328 (DeCode) e projeto Mobilizador NORTE-01-0247-FEDER-024479: ValorNatural. info:eu-repo/semantics/publishedVersion

The extensive use of synthetic additives by the food industry, some of them recognized as carcinogenic substances, has alerted consumers towards the need of adopting healthier habits. Thus, their appetence for functional foods, i.e. foods promoting positive effects on health in addition to their basic nutritional purposes, is progressively increasing. Moreover the food industry has been under high pressure due to legislation restrictions concerning the use of some of these artificial additives. Mushroom extracts contain bioactive compounds potentially useful to functionalize foodstuffs. In fact, functional foods are integrating the daily diet of consumers and are gaining prominence worldwide. Agaricus bisporus is the species with the highest consumption worldwide, showing a great potential to be used to enrich food matrices, mainly due to its high amount of ergosterol. In the present work, alcoholic extracts of A. bisporus were studied for their bioactivity and viability as functional ingredients in a food product with high water content (yogurt). Extracts were microencapsulated by spray-drying (to improve their stability and hydrophilicity), using maltodextrin crosslinked with citric acid as the encapsulating material. The effect of thermal treatment (after atomization) on crosslinking and bioactivity of the produced microspheres was tested. The incorporation of free and thermally untreated forms resulted in yogurts with higher initial antioxidant activity (EC50 values: 214 and 272 mg/mL) that decreased after 7 days (EC50 values: 248 and 314 mg/mL). Contrarily, thermally treated microencapsulated extracts showed higher antioxidant activity after the same period (EC50 values, 0 days: 106 mg/mL; 7 days: 48.7 mg/mL), in result of an effective protection provided by microencapsulation with crosslinked maltodextrin and citric acid. Functionalized yogurts showed an overall maintenance of nutritional properties. info:eu-repo/semantics/publishedVersion

Irradiation is a food processing technique supported by several globally recognized organizations (FAO, WHO, IAEA), being suitable for several food matrices, prolonging not only the shelf life of the food, but also providing microbiological decontamination. This work aimed to evaluate the effects of gamma radiation on the phenolic composition of Aloysia citrodora L. and Mentha x piperita L. infusions. Gamma radiation treatment was performed in a 60Co chamber, applying doses between 1 and 10 kGy and the results were compared with the control sample (non-irradiated, 0 kGy). The phenolic profile was determined and identified by HPLC-DAD-ESI/MS. Radiation treatment (at a dose of 10 kGy) caused statistically significant effects on the phenolic profile in both infusions, evidencing an increase in most of detected compounds, namely the majority ones. The same irradiation dose also induced an increase on the content of total phenolic acids and total phenolic compounds [1]. The results of this study can contribute to a wider knowledge of the effects of gamma irradiation on relevant bioactive compounds of several aromatic and medicinal plants, being recommended its application in these matrices. The authors are grateful FCT, Portugal and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013) and C2TN (UID/Multi/04349/2013); and to FEDER Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion