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- Sea knotgrass (Polygonum maritimum L.) as a potential source of innovative industrial products for skincare applicationsPublication . Rodrigues, Maria Joao; Matkowski, Adam; Slusarczyk, Sylwester; Magne, Christian; Poleze, Thatyana; Pereira, Catarina Guerreiro; L, CustódioSeveral Polygonum species and related genera display diverse skincare properties and were considered as active cosmetic ingredients. Thus, this work explored the halophyte sea knotgrass (Polygonum maritimum L.) as source of phenolics and flavonoids-enriched extracts with skincare properties, for the cosmetic industry. To improve the content in these compounds, different extraction solvents and methodologies were used, and the resulting extracts were tested for total contents of phenolics (TPC) and flavonoids (TFC). The acetone extract made on an ultrasound bath for 30 min had the highest TPC (275 mg of gallic acid equivalents [GAE]/g of dry weight [DW]), TFC (48.0 mg of rutin equivalents [RE]/g DW), and yield (20.3%). This extract was selected and evaluated for its in vitro antioxidant (total antioxidant, superoxide radical-scavenging and lipid peroxidation), anti-inflammatory (nitric oxide [NO] reduction on lipopolysaccharide [LPS]-stimulated RAW 264.7 macrophages), anti-wrinkles (elastase), anti-acne (lipase), antimicrobial (Escherichia colt Staphylococcus aureus, Pseudomonas aeruginosa, Candida cdbicctris), anti-melanogenic (anti-tyrosinase and reduction of melanin production on B16 4A5 melanoma cells). The extract was also appraised for toxicity, and its chemical profile was determined by ultra-high-resolution mass spectrometry (UHRMS). The acetone extract showed a high O-2(-center dot) scavenging (half maximal inhibitory concentration [IC50] = 40.4 mu g/mL), total antioxidant capacity (TAC
- In vitro enzyme inhibitory and anti-oxidant properties, cytotoxicity and chemical composition of the halophyte Malcolmia littorea (L.) R.Br. (Brassicaceae)Publication . Castañeda-Loaiza, Viana; Placines, Chloé; Rodrigues, Maria Joao; Pereira, Catarina; Zengin, Gokhan; Neng, Nuno R.; Nogueira, José M. F.; Custódio, LuísaThis work reports for the first time the in vitro anti-oxidant (towards DPPH, ABTS, copper and iron), enzymatic inhibitory (on AChE, BuChE, α-glucosidase, α-amylase and tyrosinase), cytotoxicity (towards HepG2 and HEK 293 cells), and metabolomics (by HPLC-MS) of extracts from organs of Malcolmia littorea (L.) R.Br. Extracts were constituted mainly by phenolic acids and flavonoids, and main compounds were salicylic acid and luteolin-7-O-glucoside. Samples showed reduced radical scavenging and metal chelating capacity, and only the methanol extracts reduced iron. The root's ethanol and methanol extracts, and the aerial organ's ethanol extract exhibited the highest AChE inhibition. The root's ethanol extract displayed dual anti-cholinesterase activity. Samples showed a low capacity to inhibit α-amylase, but a high α-glucosidase inhibition was obtained with the root's and flower's ethanol extracts, and flower's methanol extract. Overall, samples displayed a high inhibition against tyrosinase, reduced HepG2 cellular viability and were less toxic towards HEK 293 cells.
- Combination of hyaluronic acid and PLGA particles as hybrid systems for viscosupplementation in osteoarthritisPublication . Mota, Ana Henriques; Direito, Rosa; Carrasco, Marta P.; Rijo, Patricia; Ascensao, Lia; Viana, Ana Silveira; Rocha, Joao; Eduardo-Figueira, Maria; Rodrigues, Maria Joao; Custódio, Luísa; Kuplennik, Nataliya; Sosnik, Alejandro; Almeida, Antonio Jose; Gaspar, Maria Manuela; Reis, Catarina PintoHyaluronic acid (HA) is commonly used through intra-articular administration for viscosupplementation in osteoarthritis and other disorders. HA is generally supplied as an injection commonly reported as painful, with strong limitations after treatment. In this study, an alternative delivery system was constructed based on HA hydrogel and poly(lactic-co-glycolic acid) (PLGA) particles with oleic acid. Development studies included the determination of particle toxicity, hemolytic activity, in vitro and in vivo anti-inflammatory activity using macrophages and a murine model, respectively. This study showed that empty PLGA particles presented a mean size of 373 nm, while particles containing HA and oleic acid showed a marked particle size increase. The HA association efficiency was of 73.6% and 86.2% for PLGA particles without and with oleic acid, respectively. The in vitro HA release from PLGA particles revealed a sustained profile. Particles showed a good in vitro cell compatibility and the risk of hemolysis was less < 1%, ensuring their safety. The in vivo anti-inflammatory study showed a higher inhibition for HA-loaded PLGA particles when compared to HA solution (78% versus 60%) and they were not different from the positive control, clearly suggesting that this formulation may be a promising alternative to the current HA commercial dosage form.
- Phytochemical characterization and bioactivities of five Apiaceae species: Natural sources for novel ingredientsPublication . Zengin, Gokhan; Mahomoodally, Mohamad Fawzi; Paksoy, Mehmet Yavuz; Picot-Allain, Carene; Glamocilja, Jasmina; Sokovic, Marina; Diuzheva, Alina; Jeko, Jozsef; Cziaky, Zoltan; Rodrigues, Maria Joao; Sinan, Kouadio Ibrahime; Custódio, LuísaSeveral species of the Apiaceae family have been employed in traditional cultures for their curative virtues. The present study focused on five Apiaceae species, (Falcaria vulgaris (FV), Smyrniopsis aucher( (SA), Smyrniopsis mtutzurdagensis (SM), Smyrnium cordifolium (SC), and Actinolema macrolema (AM)). The antioxidant, enzyme inhibitory (alpha-amylase, alpha-glucosidase, acetyl- and butyrylcholinesterase, lipase, and tyrosinase), antimicrobial, phytochemical, and cytotoxicity profiles of the methanol extracts of the selected Apiaceae species were determined. SC extract (35.68 mg gallic acid equivalent/g extract) possessed the highest phenolic content while the AM extract (56.79 mg rutin equivalent/g extract) had the highest flavonoid content. HPLC-ESI-MS (High performance liquid chromatography-electrospray tandem mass spectrometry) analyses showed presence of ferulic acid in all the five species. SC extract exhibited high radical scavenging (59.28 and 94.31 mg Trolox equivalent [TE]/g extract, DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid), respectively) and reducing activity (161.44 and 113.62 mg TE/g extract, for CUPRAC (cupric reducing antioxidant capacity) and FRAP (ferric reducing antioxidant power), respectively). SM extract exhibited the highest cholinesterase's inhibitory action (3.82 and 4.76 mg galantamine equivalent/g extract, for acetyland butyrylcholinesterase, respectively). The extracts showed higher inhibition against alpha-glucosidase (7.32-11.99 mmol acarbose equivalent [ACAE]/g extract) compared to alpha-amylase (0.51-0.55 mmol ACAE/g extract). SC extract was the most active (137.54 mg kojic acid equivalent/g extract) tyrosinase inhibitor and FV extract (113.75 mg Orlistat equivalent/g) the best lipase inhibitor. SM extract showed potent antibacterial effect against B. cereus (MIC (minimum inhibitory concentration) 0.180 mg/mL), P. mirabilis (MIC 0.180 mg/mL), M. flavus (MIC 0.560 mg/mL), P. aeruginosa (MIC 0.275 mg/mL), and S. typhimurium (MIC 1.500 mg/mL). FV extract (MIC 0.140 mg/mL) suppressed A. fumigates growth. Cytotoxicity was assessed on murine macrophage (RAW 264.7), human embryonic kidney (HEK 293), and human hepatocellular carcinoma (HepG2) cell lines. FV (60.3%) and SM (57.4%) showed the highest reduction on RAW 264.7 cellular viability, whereas SM (74.1%) showed toxicity against HepG2. This study supports that the Apiaceae species could be considered as promising candidates for the development of novel pharmacophores for the management of several human ailments.
- First report of the nutritional profile and antioxidant potential of Holothuria arguinensis, a new resource for aquaculture in EuropePublication . Roggatz, Christina C.; Gonzalez-Wanguemert, Mercedes; Pereira, Hugo; Rodrigues, Maria Joao; F. G. M. Silva, Manuela; Barreira, Luísa; Varela, João; Custódio, LuísaThis work reports for the first time the nutritional profile and antioxidant potential of the edible sea cucumber Holothuria arguinensis from the North-eastern Atlantic. H. arguinensis has high levels of protein, with the amino acids profile dominated by alanine, glycine and proline and low lysine/arginine ratios. Its carbohydrate and energetic contents are also low as well as the total lipid levels, although its lipid profile is rich in polyunsaturated fatty acids (PUFA), especially arachidonic, eicosapentaenoic and docosahexaenoic acids. In addition, H. arguinensis has high levels of calcium. The water and ethanol extracts show ability to scavenge free radicals and to chelate copper and iron ions. Our results indicate that H. arguinensis has a balanced nutritional quality suitable for human consumption. In addition, it contains compounds with antioxidant potential; thus its intake can contribute for a healthy and well-balanced diet.
- Chemical profile, antioxidant, antimicrobial, enzyme inhibitory, and cytotoxicity of seven Apiaceae species from Turkey: a comparative studyPublication . Zengin, Gokhan; Sinan, Kouadio Ibrahime; Ak, Gunes; Mahomoodally, Mohamad Fawzi; Paksoy, Mehmet Yavuz; Picot-Allain, Carene; Glamocilja, Jasmina; Sokovic, Marina; Jeko, Jozsef; Cziaky, Zoltan; Rodrigues, Maria Joao; Pereira, Catarina; Custódio, LuísaSeveral Apiaceae species, used as both food and in complementary and alternative medicine, represents a rich source of potential valuable phytopharmaceuticals which necessitates scientific contemplation. In the present study, the antioxidant, enzyme inhibitory, antimicrobial, and cytotoxic properties of methanol extracts of seven Apiaceae species, (Chaerophyllum macrospermum (Willd. ex Spreng.) Fisch. & C.A.Mey. ex Hohen, Ferula rigidula Fisch. ex DC., Ferula orientalis L., Prangos ferulacea Lindl., Prangos peucedanifolia Fenzl., Ferulago setifolia K. Koch, and Pimpinella anthriscoides Boiss.) were evaluated. Species belonging to the Prangos genus exhibited the highest total phenolic content, namely P. peucedanifolia and P. ferulacea, with values of 47.90 and 44.44 mg gallic acid equivalent/g extract, respectively. P. peucedanifolia also displayed the highest radical scavenging capacity (81.53 and 102.70 mg Trolox equivalent [TE]/g extract for 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), respectively) and reducing power (165.87 and 100.09 mg TE/g extract for cupric reducing antioxidant capacity (CUPRAC) and ferric reducing antioxidant power (FRAP), respectively). C. macrospermum showed the most potent inhibition against Alzheimer's disease related enzymes, namely acetylcholinesterase (4.53 mg galantamine equivalent [GALAE]/g extract) and butyrylcholinesterase (3.22 mg GALAE/g extract). P. ferulacea (131.94 mg kojic acid (KAE) equivalent/g extract) and P. peucedanifolia (4.97 mmol acarbose equivalent (ACAE)/g extract) were potent inhibitors of tyrosinase and a-glucosidase, respectively. In general, studied species were able to reduce cellular viabilities. P. peucedanifolia possessed promising antibacterial potential against Bacillus cereus (Minimum inhibition concentration (MIC): 0.37 mg/mL), L. monocytogenes (MIC: 0.56 mg/mL), P. aeruginosa and Escherichia coli (MIC: 0.27 mg/mL), Salmonella typhimurium and Enterobacter cloacae (MIC: 0.75 mg/mL). F. rigidula showed the highest antifungal effect against Aspergillus ochraceus and Trichoderma viride (MIC: 0.10 mg/mL). The present findings could be the scientific starting point towards the pharmaceutical and/or commercial utilization of these Apiaceae species.
- The irrigation salinity and harvesting affect the growth, chemical profile and biological activities of Polygonum maritimum L.Publication . Rodrigues, Maria Joao; Monteiro, Ivo; Placines, Chloé; Castañeda-Loaiza, Viana; Slusarczyk, Sylwester; Matkowski, Adam; Pereira, Catarina; Pousao-Ferreira, Pedro; Custódio, LuísaPreviously, our group identified the halophyte Polygonum maritimum L. (sea knotgrass) as a promising source of ingredients for the cosmetic, food, pharmaceutical and veterinarian industries. To further explore this species, and to guarantee the supply of biomass for commercial purposes, it is necessary to ensure its sustainable production while assuring the preservation of its chemical and biological properties. In this context, this work aimed to cultivate this species in greenhouse conditions and to determine the influence of specific agronomic conditions, namely irrigation salinity and harvest, on the plant's growth performance and biological properties of obtained biomass. For that purpose, plants were grown in a greenhouse and irrigated with water with different salinities (0, 100, 200, 300 and 600 mM of sodium chloride - NaCl). After six weeks of irrigation, plants were cut 7 cm above soil level and submitted to an additional two harvests with a six-week interval. Plant growth performance was evaluated in terms of plant height, leaf number and surface area, moisture, and productivity. Acetone extracts were prepared from aboveground organs and evaluated for chemical composition (by spectrophotometric methods, and by ultra-high-resolution mass spectrometry - UHRMS), and for in vitro antioxidant properties [radical-scavenging activity (RSA) on DPPH and ABTS, ferric reducing antioxidant power (FRAP) and metal chelating activity on iron (ICA) and copper (CCA)]. Extracts were also appraised for in vitro anti-inflammatory activity on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Increased salinity and multiple harvests reduced plant growth and yields; the highest productivity was obtained in plants irrigated with freshwater at 2nd harvest (346 g/m(2) of dried biomass). The extracts maintained the in vitro biological properties and interesting chemical profile, however, these depended on the irrigation salinity and harvest regime. The highest antioxidant activities were obtained in extracts from plants irrigated with freshwater at the 3rd harvest (DPPH: 96.2%; ABTS: 89.1%; CCA: 61.6%; FRAP: 136%). The extracts from biomass obtained in 1st the harvest of freshwater-irrigated plants, and from those treated with 100 and 200 mM of NaCl, had significant anti-inflammatory properties. The main compounds detected were mostly flavonols (myricetin and quercetin glycosides), which varied according to both irrigation salinity and harvest. Our results indicate that sea knotgrass can be cultivated in greenhouse conditions aiming industrial commercial applications, irrigated with freshwater or with irrigation solutions with moderate salinity. Moreover, produced biomass maintain the biological and chemical properties previously detected in plants collected from the wild.
- Exploring Caralluma europaea (Guss.) NEBr. as a potential source of bioactive molecules: In vitro antioxidant and antidiabetic properties, and phenolic profile of crude extracts and fractionsPublication . Dra, Loubna Ait; Rodrigues, Maria Joao; Neng, Nuno da Rosa; Nogueira, Jose M. F.; Elamine, Youssef; Aghraz, Abdellah; Markouk, Mohamed; Larhsini, Mustapha; Custódio, LuísaCaralluma europaea (Guss.) N.E.Br is a succulent medicinal species used in the Moroccan traditional medicine for its anticancer and antidiabetic properties. In this work, this species was explored for the in vitro antidiabetic and antioxidant properties of crude methanol and dichloromethane extracts, and obtained fractions, from its aerial organs. In vitro antidiabetic properties were evaluated towards baker's yeast and rat's intestinal alpha-glucosidase, and alpha-amylase. Four in vitro methods were used to appraise the antioxidant potential, namely radical scavenging activity (RSA) on 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), and iron and copper chelating assays. Finally, the profile of the main phenolic compounds present in the samples was assessed by high-performance liquid chromatography equipped with diodearray detection (HPLC-DAD). The highest inhibition of baker's yeast alpha-glucosidase was obtained with the methanol extract (87%), and its fractions, with a maximum inhibition of 83% at the concentration of 10 mg/ml. The most significant inhibition of mammalian alpha-glucosidase was obtained with the dichloromethane crude extract (85%), and corresponding fractions at 10 mg/ml. Moderate activity was achieved with the two extracts against a-amylase (52-53%) at 10 mg/ml, but no relevant activity was detected for most fractions. Crude extracts had a significant RSA against DPPH and ABTS, with range values of 83-98% (methanol extracts) and 53-80% (dichloromethane extracts). Fractions from the methanol extract were more active against tested radicals. Crude extracts had chelating activity on both metals, and the highest copper chelation was achieved with the methanol extract, while the dichloromethane extract and the majority of fractions from both extracts presented the highest iron chelation at 10 mg/ml. A total of 21 phenolic compounds were identified and quantified by HPLC-DAD in the samples, and the main compounds detected were catechine hydrate and salicylic and caffeic acids. Our results suggest that C. europaea extracts contain compounds with in vitro antioxidant and antidiabetic properties with potential applications in the pharmaceutical and food industries.
- A comparative evaluation of biological activities and bioactive compounds of the seagrasses Zostera marina and Zostera noltei from southern PortugalPublication . Custódio, Luísa; Laukaityte, Simona; Engelen, Aschwin; Rodrigues, Maria Joao; Pereira, Hugo; Vizetto-Duarte, C; Barreira, Luísa; Rodriguez, Hortensia; Albericio, Fernando; Varela, JoãoThis work assessed the antioxidant potential, acetylcholinesterase (AChE) inhibition and the in vitro cytotoxic activity of extracts of the seagrasses Zostera marina and Zostera noltei collected from southern Portugal. The total phenolic contents (TPCs), the rosmarinic acid (RA) concentration (HPLC/DAD) and the fatty acid (FA) profile (GC/MS) are also described. Z. marina had the highest TPC, radical scavenging activity against DPPH radicals and copper chelating activity. Z. noltei had metal chelation capacity to copper and iron ions. None of the species was able to inhibit AChE. Both seagrasses had high levels of polyunsaturated FAs. Z. marina significantly and selectively reduced the viability of tumorous neuronal cells. Z. noltei was highly toxic for the three cell lines tested and was selective against hepatocarcinoma cells at the concentration of 100g/mL. RA was the main compound identified in Z. marina, but not in Z. noltei.
- Seasonal variations of the nutritive value and phytotherapeutic potential of Cladium mariscus L. (Pohl.) targeting ruminant’s productionPublication . Oliveira, Marta; Rodrigues, Maria Joao; Neng, Nuno R.; Nogueira, José Manuel Florêncio; Bessa, Rui J. B.; Custódio, LuísaIn our endeavor to identify salt-tolerant plants with potential veterinary uses in ruminants' production strategies, we focused on Cladium mariscus L. Pohl (sawgrass), due to its high total phenolic and tannin content, anti-radical properties, and ethnomedicinal uses. Aerial parts were collected along the year in Southern Portugal and evaluated for the nutritional profile and in vitro organic matter digestibility (IVOMD), aiming for its use as feed. Acetone extracts were appraised for total contents in phenolics (TPC), flavonoids (TFC), and tannins (CTC), as well as the chemical composition by HPLC-DAD and in vitro antioxidant and anti-inflammatory properties, targeting its exploitation as phytotherapeutic products. Sawgrass biomass has a limited nutritive value, due to its high neutral detergent fiber (NDF; 596-690 g kg-1 dry matter (DM)) and acid detergent fiber (ADF; 330-418 g kg-1 DM) contents, low crude protein (51.8-87.3 g kg-1 DM) and IVOMD (172-317 g kg-1 organic matter (OM)). Despite differences among seasons, the mineral profile was adequate. The extracts were rich in TPC (88-112 mg g-1), CTC (115-169 mg g-1), and TFC (18.5-20.2 mg g-1), and displayed significant antioxidant capacity, particularly in summer and autumn, whilst no seasonal influence was detected for anti-inflammatory properties (30% reduction of nitric oxide production). Eleven phenolics were quantified: chlorogenic, ferulic, and syringic acids were the most abundant, especially in the autumn sample. Overall, despite the low nutritional interest, sawgrass extracts hold the potential as a source of antioxidant and anti-inflammatory phenolic compounds.