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A. leucotrichus (Coss. & Dur.) Coss. & Dur., known in Algeria as “Kammûnes-sofi”, is a medicinal plant that finds culinary use by indigenous populations. Among others, it is used against stomach pain, indigestion, diarrhea, vomiting, fever, and to combat high blood pressure. In this work, the essential oil of A. leucotrichus, obtained by steam distillation (3h) from fruits collected in March 2015 from Tassili n'Ajjer, a vast plateau in south-east Algeria (25°30'0" N and 9°0'0" E), was chemically and biologically characterized and thereafter microencapsulated using a chitosan/TPP/vanillin system. A. leucotrichus essential oil microparticles were produced using an atomization/coagulation technique with chitosan as the shell material, sodium tripolyphosphaste (TPP) and vanillin as crosslinking agents. Comparatively to the most used chemical systems, this one presents several advantages since all the raw materials are nontoxic and no organic solvents are required. Moreover, the used microencapsulation process allows the microparticles production in a single step, without having the constrains of the traditionally used oil-in-water (o/w) emulsion based techniques. The adopted procedure comprises the following stages: (1) Chitosan solution (CS) preparation (3.0%, w/v) in acidic medium (acetic acid 3%, v/v); (2) Oil-in-water (o/w) emulsion preparation by emulsifying the essential oil (O) with the chitosan solution at O/CS ratio of 0.025 (v/v) with Tween 80 (emulsifier of HLB=15.0, 1.5%, w/v). The emulsion was homogenized at 11000 rpm during 5 min with a CAT Unidrive X homogenizer; (3) Atomization of the o/w emulsion in a Nisco VarJ30 system (flow rate: 0.3 ml/min) under pressurized nitrogen; (4) Coagulation with TPP (10%, w/v at pH 6.0) followed by vanillin crosslinking (1.0% (w/v), 50ºC at 0.5 ml/min during 2 h). Microparticles were recovered by filtration under reduced pressure, washed with distilled water and stored in the hydrated form. info:eu-repo/semantics/publishedVersion

The limitations of current cancer therapies prompt the urgent need for a more effective therapeutic strategy [1]. Graphene-based magnetic nanoparticles (GbMNPs) due to their unique properties, such as high chemical and thermal stability, high charge carrier mobility, large surface area for functionalization and superparamagnetic properties, have the potentiality to be used as efficient multifunctional nanocarrier systems [2]. However, one of the challenges of GbMNPs in biomedical applications is their tendency for agglomeration or precipitation in electrolyte solutions, such as those of body fluids [3]. To overcome this drawback, the developed GbMNPs were grafted with copolymer Pluronic F127 (PF127), yielding the materials denoted as GbMNPs@PF127. PF127 is a water-soluble and biocompatible triblock copolymer (PEO100-PPO65-PEO100) approved by the U.S. Food and Drug Administration (FDA) for use as food additive and in pharmaceutical formulations [4]. This grafting strategy allows the incorporation of a hydrophilic corona that reduces the aggregation of nanoparticles and the adsorption of blood proteins [4, 5]. Also, increases the biocompatibility of the nanosystems and its colloidal stability, prolonging blood circulation [5]. In this study, GbMNPs@PF127 were covalently conjugated with Doxorubicin (DOX), a highly effective chemotherapeutic drug against a broad spectrum of cancers. The developed therapeutic nanosystems were characterized and investigated to be used as multifunctional nanocarriers to combine thermo-chemotherapy, revealing exceptional features, such as: (i) high loading of the chemotherapeutic drug DOX; (ii) high pH stimuli-responsive controlled release; (iii) high heating efficiency profile under AMF with thermo-responsive drug release; as well as (iv) good haem- and biocompatibility even at high concentrations. The presented strategy and findings can represent a new way to design and synthesize highly stable graphene-based materials with novel structures for synergetic thermo-chemotherapy triggered by the abnormal cell microenvironment for a more effective treatment of cancer. This work was financially supported by: Project POCI-01-0145-FEDER-006984 Associate Laboratory LSRE-LCM funded by FEDER through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI) - and by nationalfunds through FCT - Fundação para a Ciência e Tecnologia R.O.R. acknowledgs the PHD scholarship SFRH/BD/97658/2013 granted by FCT. A.M.T.S. acknowledges the FCT Investigator 2013 Programme (IF/01501/2013), with financing from the European Social Fund and the Human Potential Operational Programme. info:eu-repo/semantics/publishedVersion

Discovery of new drugs from natural products is becoming widespread and an example of a promising plant is the bacupari-anão or bacupari mirim that belongs to the species: Garcinia brasiliensi Mart, family Myrtaceae. This tree is native to the forests of the Amazonian and also to the Atlantic Forest. Since ancient time, the leaves of this tree have been reported to have numerous functionalities, correlated with the presence of bioactive compounds [1]. Thus, this study aimed to evaluate the bioactive properties of G. brasiliensi leaves regarding to its possible antioxidant activity and cytotoxic properties in human tumor cells. Three different extracts of the leaves were tested: hexane, dichloromethane and ethyl acetate. Furthermore, the most active extract was characterized in terms of its phenolic compounds content.Chemical profile of the extracts was obtained using an HPLC system coupled to a diode array detector (DAD) and mass spectrometry (MS) with an electrospray ionization interface (ESI). The antioxidant activity was evaluated by four in vitro assays:DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity, reducing power, inhibition of β-carotene bleaching and inhibition of lipid peroxidation by the thiobarbituric acid reactive substances (TBARS) assay.The cytotoxicity was tested in MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung carcinoma), HeLa (cervical carcinoma) and HepG2 (hepatocellular carcinoma) cell lines and in a non-tumor primary culture (porcine liver primary culture PLP2). Among all the tested samples, ethyl acetate extract presented the highest DPPH scavenging activity (EC50 value = 31,2 ± 0,2μg/mL), reducing power (68,8± 0,2 μg/mL), β-carotene bleaching inhibition capacity (15,9 ± 0,3μg/mL) and TBARS (4,6 ± 0,2 μg/mL). It was also able to inhibit all the tested human tumor cells and none of the samples revealed toxicity for the non-tumor cell line PLP2 (GI50

The global cosmetic industry is constantly in search for bio based ingredients with less toxic effects but providing multifunctional properties, maintaining the structure and function of the skin. This will, thereby, reduce the presence of synthetic ingredients in personal care products (Taofiq, 2017). Cosmeceuticals are the newest trend in the design of personal care products, presenting medical drug-like benefits through the supply of appropriate ingredients for the maintenance of a healthy skin. They include bioactive compounds such as phytonutrients, microbial metabolites, minerals, vitamins and animal proteins (Taofiq, 2016a). Mushrooms are widely consumed due to their rich nutritional composition with recognized health promoting benefits. They contain several bioactive compounds such as polysaccharides, phenolics, terpenoids, nucleotides, steroids, fatty acids, proteins and glycol-peptides that are responsible for their antioxidant, antitumor, anti-inflammatory, antimicrobial anti-tyrosinase and other biological activities. Because of all the above-mentioned properties, there is an increasing interest in incorporating mushroom extracts and/or their isolated compounds in cosmetic formulations, expecting positive benefits on the skin. These extracts/compounds can act either as antioxidants, preservatives or as hyperpigmentation correcting agents (Taofiq, 2016a). In a general way, the use of natural-derived extracts, including mushroom extracts, as cosmeceutical ingredients can present restrictions related with the lack of stability and ease degradation of some of their metabolites. To overcome these challenges, microencapsulation techniques have been utilised taking advantage of the protection conferred by polymeric materials, particularly natural-derived and biodegradable ones. This ensures protection against oxidation and degradation, controlled or target oriented release, guaranteeing that the bioactives effectively cross the biological membranes. The specific objective of the present work was to obtain mushroom extracts-based ingredients for cosmeceutical applications. Soxhlet ethanolic extracts were prepared from Agaricus bisporus (J.E.Lange) Imbach (A) and Pleurotus ostreatus (Jacq. ex Fr.) P.Kumm (P), thereafter chemically characterised in terms of phenolic acids and ergosterol contents, and screened for their anti-inflammatory, antimicrobial, antioxidant, and anti-tyrosinase activities. The extracts were then microencapsulated in calcium alginate particles by an atomization/coagulation technique. The produced microparticles were characterised (morphology, particle size, encapsulation efficiency (EE%)) for subsequent incorporation in a base cosmetic cream. The developed formulations were evaluated for their bioactive properties, pH and colour evolution over a 6 months’ period to ascertain the stability of the bioactives and monitor their in vitro release behaviour. The composition of the bioactive compounds in the creams was also monitored by HPLC. info:eu-repo/semantics/publishedVersion

Atualmente, a indústria alimentar interessa-se pela substituição de aditivos artificiais por ingredientes naturais. Alguns extratos vegetais têm surgido como possíveis alternativas aos conservantes artificiais, nomeadamente antioxidantes [1]. De facto, têm sido desenvolvidos produtos lácteos, cárneos e de panificação, incorporando extratos de plantas aromáticas, especiarias ou frutos, que apresentam propriedades antioxidantes [2-4]. Neste trabalho, pretendeu-se comprovar a eficácia de um extrato rico em ácido rosmarínico como conservante natural de cupcakes, comparativamente a um aditivo artificial (sorbato de potássio, E202). O extrato foi obtido a partir de Melissa officinalis L. (cidreira) pela técnica de ultrassons utilizando uma mistura de etanol:água como solvente de extração. Após comprovar as suas propriedades antioxidantes (efeito captador de radicais livres, EC50 = 79 ± 2 μg/mL; poder redutor EC50 = 49 ± 1 μg/mL), antimicrobianas (contra 8 bactérias e 8 fungos contaminantes alimentares) e ausência de toxicidade (em linhas celulares), procedeu-se à sua incorporação nos cupcakes. O seu efeito foi comparado com o E202, imediatamente após incorporação e ao longo de 3 e 5 dias de armazenamento no escuro à temperatura ambiente. Todas as amostras foram analisados cromatograficamente em termos de ácidos gordos (GC-FID) e açúcares livres (HPLC-RI). Num total de 21 ácidos gordos identificados, os saturados predominaram sobre os insaturados em todas as amostras de cupcakes, sendo o ácido palmítico e o ácido oleico os maioritários. Relativamente aos açúcares, a sacarose e a glucose foram as moléculas identificadas nas amostras, sendo a quantidade de sacarose muito mais elevada. Os resultados obtidos demonstram que o extrato rico em ácido rosmarínico tem potencial para ser utilizado como aditivo natural em produtos de pastelaria, indo de encontro à atual tendência de procura dos consumidores. FCT e FEDER pelo suporte 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, J.C.M. Barreira e R.C. Calhelha. Fundos Europeus Estruturais e de Investimento (FEEI) através do Programa Operacional Regional Norte 2020, no âmbito do Projeto NORTE-01-0145-FEDER-023289: DeCodE. Empresa Mais Ervas, Lda (Alfandega da Fé, Portugal) por fornecerem as amostras de M. officinalis. info:eu-repo/semantics/publishedVersion

Rice (Oryza sativa L.) is an important cerealcrop and a staple food for more than half of the world's populations info:eu-repo/semantics/publishedVersion

Terana coerulea (Phanerochaetaceae family) is known as the cobalt crust fungus and it is used for its antibiotic properties at the Irati’s Forest (Navarra, Spain). Previous mycochemical investigations reported the isolation of corticins A-C,[1] p-terphenyl neolignans related to the antitumoural telephoric acid.[2] In this job, from powdered dry fungi, six extracts of increasing polarity were obtained and tested for cytotoxicity against four human tumour cell lines and one non-tumour primary cell culture with the sulforhodamine B assay. From the most cytotoxic one, the EtOAc extract, we isolated and identified three p-terphenyl neolignans. One of them was previously described as corticin A by Briggs et al.,[1] whose earlier structure has been revised in this work using one- and two-dimensional NMR, HRMS, positive and negative MS/MS and its peracetyl derivative in comparison with 4’’-deoxy and 4,5-dimethoxy candidusines A.[3] The other two neolignans are new natural products, named corticins D and E. These neolignans were less cytotoxic than the EtOAc extract itself, maybe due to an aerial oxidation and degradation produced when these neolignans, with catechol moieties, are definitively purified. info:eu-repo/semantics/publishedVersion

Medicinal plants are known as being a rich source of bioactive components, which can be correlated with their biological effects. The present study focused on the valorization of the leaves of Osyris quadripartite Salzm. ex Decne. with recognized folk medicinal uses in some Africa countries, namely Algeria [1, 2]. Therefore, the aim was to perform a phytochemical characterization, regarding phenolic composition, of O. quadripartita aqueous extracts and two different organic fractions (n-butanol and ethyl acetate), by using high-performance liquid chromatography coupled to a diode array detection and electrospray ionisation mass spectrometry (HPLC-DAD–ESI/MS). The separation was achieved with a Waters Spherisorb S3 ODS-2 C18 (4.6 mm × 150 mm, 3 μm), using a gradient elution, with 0.1% formic acid in water and acetonitrile. Twenty-eight individual phenolic compounds were identified: fifteen flavan-3-ols, six flavones, four flavonols, two phenolic acids and one flavanone derivative. The ethyl acetate fraction presented the highest concentration in phenolic compounds, being (+)-catechin and procyanidin dimer B1 (EC-4,8-C) the most abundant compounds (110.5 ± 0.3 and 100.5 ± 0.3 mg/g extract, respectively). The aqueous extract and the n-butanol fraction presented similar contents of phenolic compounds, being quercetin-3-O-rutinoside the main molecule present (33.8 ± 0.1 and 17.70 ± 0.02 mg/g extract, respectively). Moreover, the phenolic profiles of all fractions and extracts presented similarities in their qualitative composition, although some differences were observed, especially in the ethyl acetate fraction, where no flavone derivatives were present. This study highlights the potential of O. quadripartita fractions, rich in phenolic compounds, to be used in food, pharmaceutical and cosmetic fields. The authors are grateful to FCT and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013), L. Barros and R. Calhelha contracts; to Interreg España-Portugal for financial support through the project 0377_Iberphenol_6_E; and to the Universities of Oran 1, Abdelhamid Ibn Badis Mostaganem and the LRZA (Algeria) for financial support of W. Rached. info:eu-repo/semantics/publishedVersion

Medicinal plants are recognized since ancient times as therapeutic agents with several beneficiai properties for human health. Herein, the phenolic profile of three formulations (infusions, pills and syrups) based on artichoke, milk thistle and borututu was analysed as well as their bioactivity, namely the antioxidant, hepatoprotective and antimicrobial activities. In what concems the phenolic composition, the studied plants proved to represent a good source of bioactive compounds, especially phenolic acids and flavonoids. Regarding artichoke and milk thistle, phenolic compounds were higher in the infusion preparations when compared to the other formulations, but in the case of borututu, the syrup revealed the highest contents. Luteolin-7-0- glucuronide was the major flavonoid found in artichoke infusion, protocatechuic acid was the main constituent of borututu infusion, and isorhamnetin-0-deoxyhexoside-O-hexoside was the most abundant compound in milk thistle syrup. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financiai support to CIMO (UIDMGR/00690/2013). info:eu-repo/semantics/publishedVersion

The cobalt crust fungus Terana coerulea (Phanerochaetaceae family) was selected for a bio-guided study after an ethnobotanical survey at the Irati's Forest (Navarra, Spain) for its local use as antibiotic. Six extracts of increasing polarity, from hexane to hot water, were obtained from powdered dry fungi and tested for cytotoxicity against four human tumour cell lines and one non-tumour primary cell culture. From the most cytotoxic, EtOAc extract, we isolated and identified three terphenyl neolignans: two of them new natural products, named corticins D and E, and one previously described as corticin A, whose earlier structure has been revised. Their structural elucidation and biological evaluation as cytotoxic agents are described. This work was financially supported by Spanish MINECO co-funded by FSE (CTQ2015-68175-R) and Portuguese Foundation for Science and Technology (FCT) through CIMO (Pest-OE/AGR/UI0690/2014) and R. C. Calhelha (SFRH/BPD/68344/2010) grant. We thank to Dr Anna M. Lithgow (NMR) and Dr César A. Raposo (MS) at Plataforma Nucleus (University of Salamanca) for running some spectra, and to Koldo Villalba for collecting the sample. info:eu-repo/semantics/publishedVersion

Medicinal plants are a source of a wide range of bioactive compounds, such as alkaloids, terpenes, steroids and phenolic compounds, which are responsible for multiple biological effects [1, 2]. In the present work, the phenolic composition and bioactive potential of the aqueous and hydroethanolic extracts of Equisetum giganteum L. and Tilia platyphyllos Scop. were evaluated. The phenolic compounds were determined using a Hewlett-Packard 1100 chromatograph, with a diode array detector coupled to a MS detector API 3200 Qtrap through an ESI source and a triple quadrupole-ion trap mass analyser, while the bioactive properties were evaluated in terms of antioxidant, anti-inflammatory, and cytotoxic activities. The hydroethanolic extracts revealed higher amounts of phenolic compounds than infusions, being the concentration of flavonoids (81% of the phenolic composition) remarkably higher than the phenolic acids content (19%), in both species and extracts. T. platyphyllos presented a higher phenolic content (50.4 ± 0.4 mg/g of hydroethanolic extract and 11.65 ± 0.05 mg/g of lyophilized infusion), than E. giganteum (21.7 ± 0.4 mg/g and 4.98 ± 0.03 mg/g, respectively). Moreover, kaempferol- O-glucoside-O-rutinoside was the most abundant flavonoid in E. giganteum extract, while protocatechuic acid and (−)-epicatechin were the most abundant phenolic acid and flavonoid, respectively, in T. platyphyllos extract. Regarding the bioactive assays, both extracts obtained from T. platyphyllos showed the highest potential and none of the extracts showed toxicity in non-tumor liver cells. These biological properties were highly correlated with its content and composition in phenolic compounds. Thus, it would be interesting to evaluate the in vivo efficacy of both plant extracts to unveil the involved modes of action and to establish effective therapeutic doses. 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 L. Barros contract; to the Interreg España-Portugal for financial support through the project 0377_Iberphenol_6_E.

A secagem de plantas medicinais (tecnologia física de processamento térmico) é um processo crucial que visa a manutenção da qualidade pós-colheita, bem como a conservação de compostos bioativos com propriedades fitoterapêuticas [1,2]. Croton urucurana Baill, conhecida popularmente no Brasil como sangra d’água, é principalmente utilizada pelas suas propriedades anti-hemorrágicas, anti-inflamatórias, anti-sépticas, cicatrizantes e pela sua ação antifúngica e entomológica [3,4]. O presente trabalho teve como objectivo verificar a influência da temperatura de secagem (40, 50, 60 e 70 °C) no perfil fenólico (obtido por HPLC-DAD-ESI/MS) de folhas, caules e casca da espécie anteriormente referida; bem como nas propriedades antioxidantes e citotóxicas dos seus extratos hidroetanólicos e aquosos (obtidos por decocção). Relativamente aos compostos fenólicos, o perfil foi muito semelhante em todas as amostras, havendo somente diferenças na quantidade de cada um dos compostos; as folhas secas a 40°C revelaram a maior concentração de compostos fenólicos. Foram identificados flavan-3-óis, flavonas, flavonóis e ácidos fenólicos (o ácido gálico foi somente detetado nos caules). A maior atividade antioxidante foi observada nos extratos hidroetanólicos de folhas e caules secos a 50 °C e de casca seca a 40 °C. Todas as amostras revelaram atividade citotóxica, sendo que os GI50 mais baixos foram também obtidos nos extratos hidroetanólicas e a temperaturas de secagem mais baixas. Com estes resultados podemos concluir que as altas temperaturas de secagem influenciam o conteúdo de compostos fenólicos e as propriedades bioativas de Sangra d’água. A adequabilidade das temperaturas de secagem para o processamento de plantas medicinais é de extrema importância para a indústria de forma a preservar as suas propriedades bioativas. Instituto Federal de Educação, Ciência e Tencologia Goiano, pelo apoio e realização deste Projeto. Fundação para a Ciência e Tecnologia (FCT) e FEDER (Programa PT2020) pelo apoio financeiro ao CIMO (UID/AGR/00690/2013) e contratos de R. Calhelha e L. Barros. Ao Projeto Interreg Espanha-Portugal (0377_Iberphenol_6_E). info:eu-repo/semantics/publishedVersion

A grande diversidade na composição química das plantas é responsável pelas suas múltiplas aplicações. Para além das propriedades bioativas, as plantas são também fonte de nutrientes e metabolitos secundários nomeadamente, pigmentos naturais que podem ser utilizados em alternativa aos corantes artificiais [1,2]. Neste trabalho, foi determinada a composição de Hibiscus sabdariffa L. em açúcares, ácidos gordos, ácidos orgânicos e tocoferóis, tendo sido também avaliado o potencial bioativo e a composição em fenólicos dos seus extratos aquoso e hidroetanólico. O perfil individual de açúcares foi determinado por HPLC-RI, os ácidos gordos por GC-FID, os ácidos orgânicos por HPLC-DAD e os tocoferóis por HPLC-fluorescência. A análise de compostos fenólicos foi efetuada por HPLC-DAD-ESI/ MS, enquanto que as propriedades bioativas foram avaliadas através de ensaios de atividade antioxidante e antimicrobiana. A hepatotoxicidade dos extratos foi também testada através de uma cultura primária de células de fígado de porco. A glucose (açúcar), o ácido málico (ácido orgânico), o α- tocoferol (tocoferol) e o ácido linoleico (ácido gordo) foram os principais constituintes nas classes correspondentes. 5-(Hidroximetil)furfural foi o composto não-antociânico mais abundante, enquanto delfinidina-3-O-sambobiósido foi a antocianina presente em maior concentração em ambos os extratos testados. Apesar de ambos os extratos de H. sabdariffa terem demonstrado atividade antioxidante e antimicrobiana, destaca-se o extrato hidroetanólico com uma maior capacidade de inibição da peroxidação lipídica em homogeneizados de células cerebrais de porco e maiores efeitos bactericidas e fungicidas. Foi também evidente que nenhum dos extratos analisados revelou hepatotoxicidade. Deste modo, a espécie H. sabdariffa revelou ser interessante não só como fonte de nutrientes, mas também de compostos bioativas e pigmentos, com enorme interesse para as indústrias alimentar, cosmética e farmacêutica. 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) e ao contrato de L. Barros. Ao programa Interreg España-Portugal pelo apoio financeiro através do projeto 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion

Ganoderma lucidum {Curtis) P. Karst. é um cogumelo medicinal muito conhecido, cuja compos1çao bioativa {compostos fenólicos, triterpenoides e polissacáridos) tem sido relacionada com efeitos farmacológicos e fisiológicos (por exemplo, propriedades imunomoduladoras, citotóxicas, anti-inflamatórias, antimicrobianas e antioxidantes). No presente trabalho, os corpos frutíferos de G. ludicum foram caracterizados quanto à sua composição química (terpenoides e triterpenos, polissacáridos e tocoferóis), sendo posteriormente avaliadas diferentes propriedades bioativas (e.g., antioxidante, anti-inflamatória, antitirosinase, citotóxica e antimicrobiana) no extrato etanólico obtido por Soxhlet. O extrato foi também caracterizado quanto à sua composição em ácidos fenólicos e triterpenoides por HPLC-DAD-ESI/MS. Por fim, o extrato bioativo foi incorporado numa formulação cosmética semi-sólida, cujas propriedades bioativas e físico-químicas foram também avaliadas. A espécie G. lucidum demonstrou ser uma boa fonte de tocoferóis (141 ± 7 (lg/100 g massa seca, ms), sobretudo pela contribuição das isoformas a e õ. Os conteúdos em terpenoides, triterpenoides e polissacáridos foram 27,2 ± 0,7 mg equiv. linalol/g ms, 5,6 ± 0,5 mg equiv. ácido ursólico/g ms e 15,4 ± 0,2 mg equiv. amido/g ms, respetivamente. O extrato etanólico apresentou uma elevada atividade antioxidante (valores de EC50 para a atividade captadora de radicais livres - 0,73 ± 0,01 mg/ml e poder redutor - 0,15 ± 0,01 mg/ml), antitirosinase (EC50 = 2,81 ± 0,01 mg/ml), citotóxica (valores de Gl 50 para as linhas celulares MCF7: 61 ± 4 (lg/ml; NCI -H460: 64 ± 3 (lg/ml; HepG2: 68,44 ± 0,08 (lg/ml; Hela: 73 ± 2 (lg/ml) e antimicrobiana (MICs até 10 mg/ml contra Escherichia coli, Enterococcus faecalis, MRSA- Staphylococcus aureus resistente à meticilina, MSSA- Staphylococcus aureus suscetível à meticilina). O extrato não exibiu toxicidade para células não tumorais {GI 50> 400 ~g/ml). A análise do extrato, realizada por HPLC-DAD-ESI/MS, revelou a presença de três ácidos fenólicos {ácidos protocatéquico, p-hidroxibenzoico e siringíco) e 26 ácidos triterpénicos, sendo os ácidos ganodéricos C2, A e H os compostos mais abundantes. A formulação cosmecêutica preparada com o extrato manteve todas as bioatividades (atividades antioxidantes, antitirosinase e antimicrobiana). Os parâmetros de cor avaliados através do sistema L* a* b* foram de 70,3 ± 0,6; 12,1 ± 0,5 e 32,9 ± 0,9, respetivamente para L (luminosidade), a (verde-vermelho) e b (azul-amarelo). A formulação apresentou um pH de 4,61 ± 0,02, que é considerado desejável para a manutenção da função de barreira da pele. Em suma, o extrato etanólico de G. lucidum incorporado em formulações tópicas revelou ter um grande potencial para garantir as diferentes funções fisiológicas da pele. No entanto, as questões relacionadas com a estabilidade, permeabilidade, eficácia e biodisponibilidade tópica serão abordadas em estudos futuros. POCI-01-0145-FEDER-006984 (LA LSRE-LCM), financiado pelo FEDER, através do POCI-COMPETE2020 e FCT; Projeto NORTE-01-0145- FEDER-000006, financiado pelo NORTE2020 sob o PT2020, através do FEDER; FCT e FEDER sob o PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2013); Projeto mobilizador ValorNatural® financiado pelos Fundos Europeus Estruturais e de Investimento (FEEI) através do Programa Operacional Regional Norte 2020. info:eu-repo/semantics/publishedVersion

A curcumina tem reconhecidas propriedades bioativas e corantes. Porém, a sua aplicação em produtos alimentares é dificultada pela sua baixa solubilidade em meios hidrofílicos. De forma a ultrapassar esta limitação, foram preparadas nanopartículas de poli(vinil-pirrolidona) (PVP, 40.000g/mol)/curcumina com base na técnica de dispersão sólida, utilizando Tween 80 como estabilizante e etanol como solvente. As nanopartículas foram caracterizadas quanto à morfologia (microscopia eletrónica de transmissão) indicando forma esférica e tamanho (dispersão dinâmica de luz) com índice de polidispersão = 0,050±0,009 e diâmetro médio em intensidade = 377±15 nm. As propriedades térmicas (análise termogravimétrica, TG), interações químicas (Espectroscopia de Infravermelho com Transformada de Fourier, FTIR). As análises por TG revelaram que as nanopartículas não apresentaram perda de humidade (não se observou nenhuma transição antes dos 100 ºC), justificada pelo facto de os pontos de ligação da PVP à água terem dado origem a interações com a curcumina na sequência da encapsulação [1]. A análise por FTIR comprovou este facto dado a ausência da banda característica a 3510 cm-1 na zona do estiramento dos OHs. As propriedades bioativas (atividade antioxidante, atividade anti-inflamatória e citotoxicidade) das nanopartículas de PVP/curcumina (NC) foram comparadas às da curcumina pura utilizada no procedimento de nanoencapsulação (PC) (Sigma Aldrich) e às de uma amostra comercial dispersável (DC) (CHR Hansen, Vegex Tumeric WS5). Os valores de EC50 (μg/mL) para a atividade antioxidante (DPPH, poder redutor, β-caroteno e TBARS) evidenciaram o seguinte perfil de atividade: PC>NC>DC, similarmente à resposta anti-inflamatória (concentração responsável por 50% da inibição da produção de óxido nítrico em comparação com o controlo negativo). Quanto à citotoxicidade, a PC revelou uma atividade superior em todas as linhas celulares, tendo os valores de GI50 (μg/mL) indicado atividade NC>DC apenas nos casos das linhas HeLa, HepG2 e PLP2 e indistinta nas MCF-7 e NCI-H460. Na etapa seguinte, as formulações PC, NC e DC foram aplicadas em iogurtes, para os quais se avaliou o perfil nutricional (humidade, cinzas, gordura, proteínas, hidratos de carbono e energia) e os parâmetros de cor (L*, a* e b*) ao longo do tempo de armazenamento (TA = 0, 7 e 15 dias, 4 ºC). Independentemente de TA, os iogurtes com DC apresentaram os teores de cinza, gordura, energia e L* superiores, enquanto os valores a* e b* foram superiores para os com PC. O teor máximo de proteína foi registado para os iogurtes com NC e o teor máximo de hidratos de carbono para as amostras controlo. Quanto a TA, as amostras armazenadas durante 7 dias caracterizaram-se por teores superiores de proteína, cinza (tal como as amostras armazenadas durante 15 dias), hidratos de carbono e energia, enquanto as amostras analisadas no dia de preparação registaram os valores mais elevados para gordura e a*. O efeito do tempo de armazenamento não foi significativo nos parâmetros L* e b*. Em síntese, e apesar de se terem observado algumas diferenças significativas nos indicadores nutricionais e parâmetros de cor, pode concluir-se que as nanopartículas de PVP/curcumina são uma boa estratégia para coloração do iogurte, não se tendo registado sem alterações relevantes do seu valor nutricional e aparência. Adicionalmente, e dado a bioatividade evidenciada, a sua utilização resultará em benefícios adicionais não conseguidos com a PC (baixa solubilidade em meio hidrofílico). FEEI através do NORTE 2020 (Projetos DeCodE (NORTE-01-0145-FEDER-023289), AIProcMat@N2020 (NORTE- 01-0145-FEDER-000006) e Mobilizador ValorNatural®); FEDER através do POCI-COMPETE2020 e FCT (POCI-01- 0145-FEDER-006984 (LSRE-LCM) e UID/AGR/00690/2013 (CIMO)). info:eu-repo/semantics/publishedVersion

O ergosterol é o esteroide mais abundante nos fungos e apresenta diversas propriedades bioativas, nomeadamente atividade hipocolesterolémica, que o tornam um agente funcionalizante interessante para a área alimentar. Contudo, este é insolúvel em água e apresenta solubilidade limitada em meios apolares, sendo a via da modificação química utilizada frequentemente para ultrapassar este problema. A cromatografia revela-se essencial no acompanhamento das reações de modificação química e na purificação dos produtos obtidos. Este trabalho teve como objetivo proceder a reações de acetilação [1], esterificação [2] e metilação [3] da molécula de ergosterol. A conversão do ergosterol no decorrer das reações foi avaliada por Cromatografia em Camada Fina (TLC, placas DC-Fertigfolien Alugram® Xtra SIL G/UV254; éter de petróleo/acetato de etilo (9:1, v/v) como eluente). A mistura obtida no final de cada reação foi diluída em éter de petróleo/acetato de etilo e aplicada numa coluna cromatográfica contendo sílica em gel (14x230 mm, Geduran® Si 60). O eluente foi recolhido e a pureza do produto monitorizada por TLC por comparação com os fatores de retenção dos reagentes iniciais. As frações contendo os produtos de interesse foram separadas e avaliadas em relação às propriedades citotóxicas em linhas tumorais (MCF-7, NCI-H460, HeLa, HepG2) e não-tumorais (PLP2). Após isolamento, os compostos foram caracterizados por ressonância magnética nuclear de protão (1H RMN, CDCl3). Estes apresentaram menor atividade citotóxica do que a molécula de ergosterol parental, em todas as linhas celulares tumorais. No entanto, ao contrário desta que apresentou toxicidade em células não-tumorais (GI50 = 89 𝜇g/mL), nenhum dos compostos sintetizados apresentou toxicidade para células normais à concentração máxima testada (400 𝜇g/mL). Assim, as moléculas obtidas por modificação química poderão ser utilizadas em matrizes alimentares lipofílicas. FEEI através do NORTE 2020 (Projetos AIProcMat@N2020 (NORTE-01-0145-FEDER-000006) e Mobilizador ValorNatural®); FEDER através do POCI-COMPETE2020 e FCT (POCI-01-0145-FEDER-006984 (LSRE-LCM), UID/AGR/00690/2013 (CIMO), bolsa SFRH/BPD/101413/2014 de S. Heleno e contrato de R. Calhelha). CAPES (Brasil) pelo apoio através do proj. 99999.000488/2016-00 (Programas Estratégicos-DRI). N/A

Rice husk, a by.product of the rice milling industry, is one of the agricultural waste materialsthat has received high attention. info:eu-repo/semantics/publishedVersion

A. leucotrichus (Coss. & Dur.) Coss. & Dur., known in Algeria as “Kammûnes-sofi”, is a medicinal plant that finds culinary use by indigenous populations. Among others, it is used against stomach pain, indigestion, diarrhea, vomiting, fever, and to combat high blood pressure. In this work, the essential oil of A. leucotrichus, obtained by steam distillation (3h) from fruits collected in March 2015 from Tassili n'Ajjer, a vast plateau in south-east Algeria (25°30'0" N and 9°0'0" E), was chemically and biologically characterized and thereafter microencapsulated using a chitosan/TPP/vanillin system. Chemical characterization allowed the identification of ten constituents representing 98.6% of the whole essential oil composition. Oxygen-containing monoterpenes (87.2 %) were found to be the main group of components, followed by monoterpene hydrocarbons (11.1 %) and oxygen-containing sesquiterpenes (0.35 %). Perilla aldehyde was identified as the main component present in the essential oil accounting for 85.6 % of the total composition. Additionally, the oil presented antioxidant (EC50 28±2 mg/ml, concentration able to scavenge 50% of DPPH radicals), anti-inflammatory (EC50 11.7±0.7 μg/ml, concentration able to inhibit 50% of NO formation) and antimicrobial (against Escherichia coli: minimum inhibitory concentration (MIC) 10 mg/ml and minimum bactericidal concentration (MBC) 10 mg/ml; against Staphylococcus aureus: MIC 20 mg/ml and MBC 20 mg/ml) activities.A. leucotrichus essential oil microparticles were produced using an atomization/coagulation technique with chitosan as the shell material, sodium tripolyphosphaste (TPP) and vanillin as crosslinking agents. Comparatively to the most used chemical systems, this one presents several advantages since all the raw materials are nontoxic and no organic solvents are required. Moreover, the used microencapsulation process allows the microparticles production in a single step, without having the constrains of the traditionally used oil-in-water (o/w) emulsion based techniques. The adopted procedure comprises the following stages: (1) Chitosan solution (CS) preparation (3.0%, w/v) in acidic medium (acetic acid 3%, v/v); (2) Oilin- water (o/w) emulsion preparation by emulsifying the essential oil (O) with the chitosan solution at O/CS ratio of 0.025 (v/v) with Tween 80 (emulsifier of HLB=15.0, 1.5%, w/v). The emulsion was homogenized at 11000 rpm during 5 min with a CAT Unidrive X homogenizer; (3) Atomization of the o/w emulsion in a Nisco VarJ30 system (flow rate: 0.3 ml/min) under pressurized nitrogen; (4) Coagulation with TPP (10%, w/v at pH 6.0) followed by vanillin crosslinking (1.0% (w/v), 50ºC at 0.5 ml/min during 2 h). Microparticles were recovered by filtration under reduced pressure, washed with distilled water and stored in the hydrated form.The produced microparticles were preliminary analyzed by optical microscopy (OM) using a Nikon eclipse 50i microscope to access size and morphology. This analysis showed the presence of spherical and individualized structures with an estimated particle size between 15 and 75 μm. Moreover, microparticles chemical structure was analyzed by FTIR, the thermal degradation was evaluated by TG and microparticle size distributions were measured by laser diffraction. The results shown the production of viable microparticles, indicating that the chitosan/ TPP/vanillin chemical system is a feasible alternative for a green A. leucotrichus essential oil encapsulation, when the atomization/coagulation technique is used. Moreover, taking into account the antimicrobial activity of A. leucotrichus essential oil, the produced microparticles can be a good alternative for cosmetic application as preservative. CIMO UID/AGR/00690/2013 and POCI-01-0145-FEDER-006984 (LA LSRE-LCM), funded by FEDER, through POCI-COMPETE2020 and FCT; Project NORTE-01-0145-FEDER-000006, funded by NORTE2020 under PT2020, through FEDER, S. A. Heleno (SFRH/BPD/101413/2014) and R.C. Calhelha contract info:eu-repo/semantics/publishedVersion

Sustainable food options are increasing among the most concerned consumers, in which they seek to combine new ingredients with potential health benefits [1]. Edible flowers provide new colors, textures and vibrancy to any dish, and apart from the "glam" factor, they can provide bioactive compounds [2]. In the presentwork, the edible petals of Rosa canina L, gently supplied by the RBR Foods Company (Portugal), were tested for their cytotoxic activity against tumor cell lines. The assays were conducted in the hydromethanolic extract and in the lyophilized infusion, being both of them re-dissolved in water in order to obtain stock solutions at 100 mg/mL for successive dilutions until determination of Glso values (concentration that inhibits 50% of the net cell growth). Four human tumor cell lines were tested: MCF-7 (breast adenocarcinoma), NCIH460 (non-small cell lung câncer), HeLa (cervical carcinoma) and HepG2 (hepatocellular carcinoma). A non-tumour primary cells' culture (assigned as PLP2) was also prepared from a freshly harvested porcine liver, and used in the bioassays. Ellipticine was used as the positive contrai. While the hydromethanolic extract inhibited the growth of HeLa (308 pg/mL), the lyophilized infusion inhibited MCF-7 (377 pg/mL), respectively. Both extracts presented cytotoxicity for HepG2 (297 |jg/mL and 315 pg/mL for hydromethanolic extract and infusion, respectively). Nane of the extracts showed toxicity towards PLP2. After chemical characterization of the extracts, flavonoids were the only phenolic compounds present, being quercetin-3-O-glucoside and kaempferol-3-O-glucoside the major compounds present in both extracts. These results showed that R. canina edible flowers are sources of bioactive compounds related with cytotoxic properties in human tumor cell lines, highlighting their applicability potential in nutraceutical and pharmaceutical fields. The authors are grateful to lhe Foundation for Science and Technology (FCT, Portugal) and FEDER under Pmgramme PT2020 for financial support to CIMO (UID/AGR/00690/2013). L. Barras (SFR/BPD/107855/2015) and M. l. Dias (SFRH/BD/84485/2012) grants. info:eu-repo/semantics/publishedVersion

Mushrooms have been used as sources of natural compounds for pharmaceutical applications. Not just fruiting bodies, but also their mycelium have been recognized as important sources of bioactive molecules. Pleurotus eryngii (DC.) Quél, in particular, is widely consumed due to its nutritional value and bioactive properties. On the other hand, Suillus bellinii (Inzenga) Watling is known as an ectomycorrhizal symbiont whose bioactivity has been scarcely studied. In the present work, each one of this species (fruiting body, mycelium and culture media) was characterized in terms of wellrecognized bioactive compounds namely, phenolic acids and ergosterol, by high performance liquid chromatography coupled to a photodiode array or an ultraviolet detector, respectively. The antioxidant activity of the samples was evaluated by four different assays: free radical scavenging activity, reducing power, b-carotene bleaching inhibition and thiobarbituric acid reactive substances (TBARS) assay. The cytotoxic activity, was evaluated either in human tumor cell lines (MCF-7- breast adenocarcinoma, NCI-H460- non-small cell lung cancer, HeLa- cervical carcinoma and HepG2- hepatocellular carcinoma), as also in a non-tumor porcine liver cells established in-house (PLP2). S. bellinii fruiting body and its mycelium showed higher contents of ergosterol and phenolic acids, and higher antioxidant activity than P. eryngii counterparts, Nevertheless, P. eryngii mycelium showed a cytotoxicity similar (sometimes superior) to its fruiting body, in comparison with S. bellinii mycelium that presented a lower anti-proliferative activity than its fruiting body. Overall, P. eryngii and S. bellinii, as well as their mycelia and culture media can be used as sources of bioactive compounds or as ingredients to be included in applications with antioxidant and cytotoxic properties. info:eu-repo/semantics/publishedVersion

A microencapsulação tem vindo a ser estudada com o objetivo de proteger substâncias sensíveis e possibilitar a adição de compostos com baixa solubilidade em matrizes alimentares. Neste trabalho, um extrato etanólico de Agaricus bísporus (Lange) Imbach foi microencapsulado por spray-drying [l] seguido de tratamento térmico, gerando micropartículas de maltodextrina reticulada com ácido cítrico. As micropartículas foram caracterizadas por Espectroscopia de Infravermelho (FTIR), Análise Termogravimétrica (TG), tamanho de partículas, eficiência de encapsulaçâo, rendimento e carga em ergosterol. Foi avaliada a atividade citotóxica numa cultura primária de células hepáticas [2] e, em seguida, as micropartículas contendo o extrato foram adicionadas ao iogurte. Os valores de eficiência de encapsulação, rendimento e carga em ergosterol foram de 40,2% (em termos de ergosterol), 35% e 16,4 mg/g, respetivamente. Os espectros de FTIR evidenciaram a reaçâo de reticulaçao entre a maltodextrina e o ácido cítrico, já que houve o aparecimento da banda relativa ao estiramento do grupo éster em 1732 cm-1. Tal banda foi intensificada após o tratamento térmico, demonstrando que a reação de reticutação continuou durante esta etapa. As análises térmicas mostraram o ganho em estabilidade consequente da reticulação da matriz polimérica. Os iogurtes contendo extrato encaDSulado em que as micropartículas passaram por tratamento térmico revelaram maior atividade antioxidante após sete dias de armazenamento. Tal pode ser atribuído à proteção resultante da reticulação da maltodextrina com o ácido cítrico durante a atomização e também durante o tratamento térmico, conforme evidenciado pelas análises de FTIR e TG. As amostras de iogurte contendo o extrato microencapsulado não apresentaram hepatotoxicidade para células não tumorais. Os resultados mostraram que a encapsulação do extrato de A bisporus é uma estratégia promissora para a proteção dos seus compostos e também para a funcionalização de alimentos. FEEI através do NORTE 2020 (Projetos AIProcMat@N2020 (NORTE-01-0145-FEDER-000006) e Mobilizador ValorNatural8 ); FEDER através do POCI-COMPETE2020 e FCT (POCI-01-0145-FEDER-006984 (LSRE-LCM), UID/AGR/00690/2013 (CIMO), contrato de R. Calhelha e SFRH/BPD/101413/2014 de S. Heleno). CAPES (Brasil) pelo apoio através do projeto 99999.000488/2016-00 (Programas Estratégicos - DRI). info:eu-repo/semantics/publishedVersion