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- 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.
- 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.
- Dataset on functional and chemical properties of the medicinal halophyte Polygonum maritimum L. under greenhouse cultivationPublication . Rodrigues, Maria Joao; Monteiro, Ivo; Placines, Chloé; Castañeda-Loaiza, Viana; Ślusarczyk, Sylwester; Matkowski, Adam; Pereira, Catarina; Pousão-Ferreira, Pedro; Custódio, LuísaThis data article includes data and analyses on the effect of different agronomic techniques on the production of Polygonum maritimum L. (sea knotgrass), namely different salinity irrigation treatments (0, 100, 200, 300 and 600 mM of NaCl) and a multi-harvest regime, and their relation with the chemical profile (ultra-high-resolution mass spectrometry - UHRMS), in vitro antioxidant [radical-scavenging activity (RSA) of DPPH and ABTS, copper chelating activity and ferric reducing antioxidant power] and anti-inflammatory (nitric oxide reduction on lipopolysaccharide-stimulated macrophages) activities. For further interpretation of the data presented in this work, please see the related research article "The irrigation salinity and harvesting affect the growth, chemical profile and biological activities of Polygonum maritimum L." (Rodrigues et al., 2019).
- Improvement of carotenoid extraction from a recently isolated, robust microalga, Tetraselmis sp. CTP4 (chlorophyta)Publication . Lisa Schueler, Lisa M. Schueler; Katkam, Dr. Gangadhar N.; Duarte, Paulo A. S.; Placines, Chloé; Molina-Márquez, Ana María; Léon-Bañares, Rosa; Sousa, Vânia S.; Varela, João; Barreira, LuísaIn recent years, there has been increasing consumer interest in carotenoids, particularly of marine sustainable origin with applications in the food, cosmeceutical, nutritional supplement and pharmaceutical industries. For instance, microalgae belonging to the genus Tetraselmis are known for their biotechnologically relevant carotenoid profile. The recently isolated marine microalgal strain Tetraselmis sp. CTP4 is a fast-growing, robust industrial strain, which has successfully been produced in 100-m3 photobioreactors. However, there are no reports on total carotenoid contents from this strain belonging to T. striata/convolutae clade. Although there are several reports on extraction methods targeting chlorophytes, extraction depends on the strength of cell coverings, solvent polarity and the nature of the targeted carotenoids. Therefore, this article evaluates different extraction methods targeting Tetraselmis sp. CTP4, a strain known to contain a mechanically resistant theca. Here, we propose a factorial experimental design to compare extraction of total carotenoids from wet and freeze-dried microalgal biomass using four different solvents (acetone, ethanol, methanol or tetrahydrofuran) in combination with two types of mechanical cell disruption (glass beads or dispersion). The extraction efficiency of the methods was assessed by pigment contents and profiles present in the extracts. Extraction of wet biomass by means of glass bead-assisted cell disruption using tetrahydrofuran yielded the highest amounts of lutein and β-carotene (622 ± 40 and 618 ± 32 µg g-1 DW, respectively). Although acetone was slightly less efficient than tetrahydrofuran, it is preferable due to its lower costs and toxicity.
- If you cannot beat them, join them: exploring the fruits of the invasive species Carpobrotus edulis (L.) NE Br as a source of bioactive productsPublication . Castañeda-Loaiza, Viana; Placines, Chloé; Rodrigues, Maria João; Pereira, Catarina; Zengin, Gokhan; Uysal, Ahmet; Jeko, Jozsef; Cziaky, Zoltan; Reis, Catarina Pinto; Gaspar, Maria Manuela; Custódio, LuísaThe halophyte species Carpobrotus edulis (L.) N.E. Br, also known as Hottentot-fig, is one of the 20 most aggressive invasive species of coastal areas worldwide. It is native to South Africa, where it is used in traditional medicine for the treatment of several diseases, including tuberculosis and acquired immunodeficiency syndrome (AIDS) caused by the human immunodeficiency virus (HIV). Aiming at a sustainable use of its biomass as a value-added product, this work reports for the first time the in vitro antioxidant, anti-microbial, enzymatic inhibitory properties and toxicity of peel and flesh extracts of Hottentot-fig mature fruits. The extracts' chemical composition was also determined by spectrophotometric methods (total contents of phenolics: TPC; flavonoids: TFC and tannins: TTC), and by high-performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS/MS). The peels' extracts had generally the highest TPC, TFC and TTC, especially the ethanol ones (TPC: 272.82 mg gallic acid equivalents (GAE)/g dry weight (DW), TFC: 1.58 mg quercetin equivalents (QE)/g DW and TTC: 20.3 mg catechin equivalents (CE)/g DW). The peels' extracts also had the highest diversity of compounds, mostly phenolic acids, flavonoids, and coumarins, as identified by HPLC-ESI-MS/MS. Some molecules were specific to a particular fruit part, for example, coumaric acid and uvaol in the peel, and vanillin and kaempferol-O-(rhamnosyl)hexosylhexoside in the flesh. Some compounds are here described for the first time in Hottentot-fig, such as azelaic acid and emodin. The peel's extracts had the highest anti radical activity, especially the ethanol and acetone towards 2,2-diphenyl-1-picrylhydrazyl (DPPH) (half maximal inhibitory concentration (IC50) values of 0.59 and 0.88 mg/mL, respectively), and the acetone extract against 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) (IC50 = 0.56 mg/mL). Samples had nil capacity to chelate iron, a low copper chelation potential, but a significant capacity to reduce iron, especially the ethanol (IC50 = 0.09 mg/mL) and the acetone extracts of peels (IC50 = 0.10 mg/mL) and flesh (IC50 = 0.11 mg/mL) and also the water peel's extracts (IC50 = 0.18 mg/mL). Samples had nil to low activity towards the enzymes acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), alpha-amylase and alpha-glucosidase, but displayed a strong inhibition of tyrosinase, especially the ethanol peel's extracts (29.55 mg kojic acid equivalents (KAE)/g). Samples had nil to low in vitro toxicity towards human keratinocytes. All together our results suggests possible novel biotechnological applications of Hottentot-fig fruits as sources of innovative bioactive ingredients for the food, cosmetic, agriculture and/or pharmaceutical industries.
- Growth performance, in vitro antioxidant properties and chemical composition of the halophyte Limonium algarvense Erben are strongly influenced by the irrigation salinityPublication . Rodrigues, Maria Joao; Monteiro, Ivo; Castañeda-Loaiza, Viana; Placines, Chloé; Conceição Oliveira, M.; Reis, Catarina; Caperta, Ana D.; Soares, Florbela; Pousão-Ferreira, Pedro; Pereira, Catarina; Custódio, LuísaLimonium algarvense Erben (sea lavender) is a halophyte species with potential to provide natural ingredients with in vitro antioxidant, anti-inflammatory, neuroprotective and antidiabetic properties. This study reports for the first time the 1) cultivation of sea lavender in greenhouse conditions under irrigation with freshwater (approx. 0 mM NaCl) and saline aquaculture wastewater (300 and 600 mM NaCl), and 2) the influence of the irrigation salinity on the plant performance (e.g growth, number of produced leaves and flowers), in vitro antioxidant properties [radical scavenging activity (DPPH and ABTS), ferric reducing antioxidant power (FRAP), metal chelating properties on copper (CCA) and iron (ICA)], toxicity (in vitro on three mammalian cell lines) and chemical composition (determined by LC-ESI-HRMS/MS). The freshwater-irrigated plants had better growth performance than those irrigated with saltwater. Extracts from wild plants, had the highest antioxidant activity, but those from cultivated ones kept high in vitro antioxidant properties and interesting chemical profile. The flowers' extracts of plants irrigated with 300 mM NaCl had the highest antioxidant activities against DPPH, whereas those from freshwater-irrigated plants were more active on ABTS, CCA and FRAP. Most of the extracts showed nil toxicity. The flowers' extracts displayed the highest diversity of compounds, mainly quercetin, apigenin, luteolin, naringenin and their glycoside derivatives. Moreover, their abundance varied with the irrigation salinity. These data indicate that sea lavender plants can be successfully cultivated in greenhouse conditions under fresh- and saltwater irrigation, maintaining interesting biological and chemical properties.