Browsing by Author "Fernandes, Eliana"
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- Antioxidant activity and in vitro inhibition of tumor cell growth by leaf extracts from the carob tree (Ceratonia siliqua)Publication . Custódio, Luísa; Fernandes, Eliana; Escapa, Ana Luísa; López-Avilés, Sandra; Fajardo, Alba; Aligué, Rosa; Alberício, Fernando; Romano, AnabelaThe methanol leaf extracts of female cultivars of the carob tree [Ceratonia siliqua L. (Fabaceae)] and of hermaphrodite and male trees were investigated for their contents of phenolic compounds, their in vitro antioxidant activity, measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging and linoleic acid system assays, and their in vitro tumor growth inhibition on HeLa cells. The different cultivars and trees showed high levels of phenols, and considerable variations in the amount of these compounds. The extracts showed significant radical scavenging activity (RSA), which was not significantly affected by the gender of the tree. From the female cultivars tested, Galhosa exhibited the highest RSA. Gender significantly affected the antioxidant activity of the extracts measured by the linoleic acid system assay, and males and hermaphrodites showed the highest activities. The extracts displayed a remarkable ability to inhibit tumor cell proliferation, and their bioactivity varied with different cultivars or trees tested. Extracts from male and hermaphrodite trees exhibited higher capacity to inhibit the proliferation of HeLa cells than the female cultivars.
- Antioxidant activity, enzyme inhibition, photoprotection, cytotoxicity, and phytochemical profiling of sea lavender (Limonium algarvense Erben) seed extracts for dermo-cosmetic usePublication . Lescano, Leonardo; Cziáky, Zoltán; Kurt-Celep, İnci; Zengin, Gökhan; Fernandes, Eliana; Trentin, Riccardo; Guerreiro Pereira, Catarina Alexandra; Custódio, Luísa; Rodrigues, Maria JoãoDespite sea lavender being a medicinal species, research on its seeds' biological properties and chemical composition is unexplored. Thus, this study evaluated the effect of different extraction solvents on the biological activities and chemical profile of greenhouse-cultivated sea lavender seeds, aiming at their potential use as a dermo-cosmetic ingredient. Therefore, ethyl acetate, acetone, ethanol, and water extracts were examined for their antioxidant activity, enzyme inhibition, photoprotection, and cytotoxicity, followed by phytochemical analysis through spectrophotometric methods, further detailed by Ultrahigh-Performance Liquid Chromatography Coupled with Electrospray Ionization Mass/Mass Spectrometry (UHPLC-Esi-MS/MS).ResultsThe water extract demonstrated significant antioxidant activity, evidenced by low half maximal effective concentration (EC50) values in scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2 '-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, reducing iron and chelating copper (296, 478, 230 and 678 mu g/mL, respectively). The ethanol extract was more effective in inhibiting cosmetic-related enzymes, particularly elastase and hyaluronidase (2.18 and 3.21 mu g/mL, respectively). The water and acetone extracts had the highest sun protection factors (23.2 and 18.9, respectively). All the extracts had nil to weak cytotoxicity (70-120% cell viability) towards mammalian cell lines. The water extract had the highest phenolics and condensed tannins (115 and 78.30 mg/g extract, respectively), while the ethanol contained the most flavonoids (62.73 mg/g extract). UHPLC-ESI-MS/MS analysis identified ethyl gallate, myricetin, rutin, and quercetin as major components of the ethanol extract, whereas myricetin-O-rutinoside isomers are predominant in the water extract.ConclusionsThese findings highlight the potential of greenhouse-cultivated sea lavender seeds as potential dermo-cosmetic ingredients, with ethanol and water extracts demonstrating superior biological activities and chemical profiles, significantly contributing to general skin health and protection.
- Antioxidant and cytotoxic activities of carob tree fruit pulps are strongly influenced by Gender and cultivarPublication . Custódio, Luísa; Fernandes, Eliana; Escapa, Ana Luísa; Fajardo, Alba; Aligué, Rosa; Alberício, Fernando; Neng, Nuno Rosa; Nogueira, José Manuel F.; Romano, AnabelaExtracts from fruit pulps of six female cultivars and two hermaphrodite Portuguese carob trees [(Ceratonia siliqua L., Fabaceae)] exhibited strong antioxidant activity and were rich in phenolic compounds. The extracts decreased the viability of different human cancer cell lines on a dose- and time-dependent manner. Gender and cultivar significantly influenced the chemical content and the biological activities of the extracts. Extracts from hermaphrodite trees had a higher content of phenolic compounds, and exhibited higher antioxidant and cytotoxic activities. Among. females, cv. Aida had the highest radical scavenging activity and total content of phenolics, Mulata the highest capacity to inhibit lipid oxidation and Gasparinha the strongest cytotoxic activity on HeLa cells. The decrease in cell viability was associated with apoptosis on HeLa and MDA-MB-231 lines. (+)-Catechin and gallic acid (GA) were the main compounds identified in the extracts, and GA contributed to the antioxidant activity. Our results show that the antioxidant and cytotoxic activities of carob tree fruit pulps are strongly influenced by gender and cultivar, and provide new knowledge about the advantages of hermaphrodite trees over female cultivars, namely, as a source of compounds with biological interest, which may represent an increase of their agronomic interest.
- Application of In Vitro plant tissue culture techniques to halophyte species: A reviewPublication . L, Custódio; Charles, Gilbert; Magné, Christian; Barba-Espín, Gregorio; Piqueras, Abel; Hernández, José A.; Ben Hamed, Karim; Castañeda-Loaiza, Viana; Fernandes, Eliana; Rodrigues, Maria JoãoHalophytes are plants able to thrive in environments characterized by severe abiotic conditions, including high salinity and high light intensity, drought/flooding, and temperature fluctuations. Several species have ethnomedicinal uses, and some are currently explored as sources of food and cosmetic ingredients. Halophytes are considered important alternative cash crops to be used in sustainable saline production systems, due to their ability to grow in saline conditions where conventional glycophyte crops cannot, such as salt-affected soils and saline irrigation water. In vitro plant tissue culture (PTC) techniques have greatly contributed to industry and agriculture in the last century by exploiting the economic potential of several commercial crop plants. The application of PTC to selected halophyte species can thus contribute for developing innovative production systems and obtaining halophyte-based bioactive products. This work aimed to put together and review for the first time the most relevant information on the application of PTC to halophytes. Several protocols were established for the micropropagation of different species. Various explant types have been used as starting materials (e.g., basal shoots and nodes, cotyledons, epicotyls, inflorescence, internodal segments, leaves, roots, rhizomes, stems, shoot tips, or zygotic embryos), involving different micropropagation techniques (e.g., node culture, direct or indirect shoot neoformation, caulogenesis, somatic embryogenesis, rooting, acclimatization, germplasm conservation and cryopreservation, and callogenesis and cell suspension cultures). In vitro systems were also used to study physiological, biochemical, and molecular processes in halophytes, such as functional and salt-tolerance studies. Thus, the application of PTC to halophytes may be used to improve their controlled multiplication and the selection of desired traits for the in vitro production of plants enriched in nutritional and functional components, as well as for the study of their resistance to salt stress.
- Comparative metabolomic and bioactivity profiling of wild and greenhouse–Transplanted Plantago coronopus LPublication . Trentin, Riccardo; Rodrigues, Maria João; Santos, Alexia dos; Fernandes, Eliana; Custódio, LuísaThis study investigates the influence of growing conditions on the metabolomic composition and associated bioactivities of Plantago coronopus L. Methanol extracts were prepared from freeze-dried biomass of both wild and greenhouse-transplanted plants and evaluated for in vitro antioxidant capacity and inhibitory effects on enzymes relevant to Alzheimer's disease (acetylcholinesterase, butyrylcholinesterase), type 2 diabetes (alpha-glucosidase, alpha-amylase), and obesity-related disorders (lipase). An untargeted metabolomic analysis was performed using mass spectral library matching and in silico annotation tools to characterise the chemical composition of the extracts. Extracts from wild specimens exhibited stronger antioxidant activity and cholinesterase inhibition, coinciding with higher levels of total phenolics, flavonoids, and carbohydrates. These differences were associated with increased levels of bioactive metabolites, including caffeic acid derivatives, terpenoids, and lipid-like compounds, possibly linked to environmental stress responses. Despite lower bioactivity, greenhouse-grown plants still produced valuable compounds such as acteoside, echinacoside, and plantamajoside. These findings highlight the potential of P. coronopus as a source of bioactive metabolites and suggest that manipulating environmental conditions in controlled cultivation could enhance phytochemical yields.
- A comparative study of the influence of soil and non-soil factors on seed germination of edible salt-tolerant speciesPublication . Castañeda-Loaiza, Viana; Rodrigues, Maria João; Fernandes, Eliana; Custódio, LuísaCultivating edible salt-tolerant plants (halophytes) for human consumption is increasingly important due to climate change and soil salinization, and offers sustainable agricultural solutions. Optimizing seed germination, the crucial initial stage of crop growth, is essential for enhancing crop production. This study aimed to optimize the germination of edible halophytes under greenhouse conditions, focusing on select soil (salinity and substrate) and non-soil-related factors (chemical and mechanical treatments). The target species were selected for their commercial value and included Mesembryanthemum crystallinum L. (crystalline iceplant), Salicornia ramosissima J. Woods (sea aspara gus), Medicago marina L. (sea medick), Ammophila arenaria (L.) Link (European beachgrass), Portulaca oleracea L. (common purslane), and Atriplex halimus L. (Mediterranean saltbush). Salinity negatively impacted germination rates (GRs) and delayed mean germination time (MGT) across species. P. oleracea had the highest GR (95.6%) in coco peat under freshwater irrigation, and the shortest MGT (5.2 days). A. halimus did not germinate under the tested conditions. Scarification with sulfuric acid improved the GR of M. marina by 42.2%, while scarification with ultrasounds improved the GR of A. arenaria by 35.5%. Our results indicate that the choice of substrate and the application of specific treatments like scarification can significantly improve the germination of certain halophyte species under variable saline conditions.
- Delving into the phytochemical constituents and biological activities of Scorzonera coriacea extracts: new perspectives from in vitro and in silico studiesPublication . Zengin, Gökhan; Ak, Gunes; Yagi, Sakina; Cetiz, Mehmet Veysi; Tutuş, Ramazan; Rodrigues, Maria João; Fernandes, Eliana; Custódio, Luísa; Yildiztugay, Evren; Aly, Shaza H.; Eldahshan, Omayma A.; Singab, Abdel Nasser B .The current study was designed to investigate the chemical composition, antioxidant, enzyme inhibitory, and cytotoxic activities of Scorzonera coriacea A.Duran & Aksoy. Both organs were rich in total phenolic content, with the highest content recorded from the 70% EtOH (48.41 mg GAE/g) and aqueous (47.11 mg GAE/g) extracts of the roots. All aerial parts extracts accumulated higher total flavonoid content than their respective roots extracts, with the highest amount found in their EtOH extract (36.44 mg RE/g). Chemical analysis revealed the presence of 86 compounds belonging to organic acids, phenolic acids, flavonoids, coumarins, anthocyanins, terpenes, saponins, and fatty acids and their derivatives, with the aerial parts accumulating the highest number. The roots displayed the strongest antiradical and ion-reducing capacities. EtOH extract of both organs recorded the highest acetylcholinesterase activity (2.77 and 3.02 mg GALAE/g; p ≥ 0.05), while that of the root showed the best butyrylcholinesterase activity (3.49mg GALAE/g) and that of the aerial parts the best tyrosinase inhibitory (59.07 mg KAE/g). EtOAc of the root exhibited the best cytotoxicity towards theHepG2 cell line (cell viability = 29.30%), butwas also toxic towards HEK293 cells (cell viability = 11.72%). In silico screening supported these findings by identifying multiple strong ligand–protein interactions. Molecular dynamics simulations further confirmed the structural stability of selected complexes. In silico profiling docked 26 phytochemicals against 14 therapeutic targets, generating 364 complexes, of which 62% showed ΔG ≤ −7.0 kcal⋅mol−1. Binding energies ranged from−1.4 to −10.7 kcal⋅mol−1, with PD-1–Eriodictyol-7-O-neohesperidoside the best. For metabolic enzymes, Eriodictyol-7-O-neohesperidoside yielded the top α-amylase score and Diosmetin-7-O-glucoside the top α-glucosidase score, while several flavonoids bound AChE/BChE strongly; in contrast, tyrosinase displayed poor affinity overall. 100-ns MD simulations on five top complexes indicated stable behavior for C1 and C4, whereas C2/C3/C5 showed loosening interactions over time.These findings showed that S. coriacea could be a promising source of bioactive compounds with potential therapeutic applications.
- A first approach for the micropropagation of the edible and medicinal Halophyte Inula crithmoides L.Publication . Rodrigues, Maria João; Castañeda-Loaiza, Viana; Fernandes, Eliana; Custódio, LuísaInula crithmoides L. (golden samphire) is an edible aromatic halophyte species with confirmed nutritional and medicinal properties attributed to the presence of important metabolites, including proteins, carotenoids, vitamins, and minerals. Therefore, this study aimed at establishing a micropropagation protocol for golden samphire that can serve as a nursery approach to its standardized commercial cultivation. For that purpose, a complete regeneration protocol was developed by improving shoot multiplication from nodal explants, rooting, and acclimatization methodologies. The treatment with BAP alone induced the maximum shoot formation (7–7.8 shoots/explant), while IAA treatment increased the shoot height (9.26–9.5 cm). Furthermore, the treatment that coupled best shoot multiplication (7.8 shoots/explant) and highest shoot height (7.58 cm) was MS medium supplemented with 0.25 mg/L BAP. Moreover, all shoots produced roots (100% rooting), and multiplication treatments did not exert significant effect on root length (7.8–9.7 cm/plantlet). Moreover, by the end of the rooting phase, plantlets cultivated with 0.25 mg/L BAP had the highest shoot number (4.2 shoots/plantlet), and plantlets from 0.6 mg/L IAA + 1 mg/L BAP presented the highest shoot height (14.2 cm) similar to control plantlets (14.0 cm). The survival up to the ex-vitro acclimatization stage was increased from 9.8% (control) to 83.3%, when plants were treated with a paraffin solution. Nevertheless, the in vitro multiplication of golden samphire is a promising way for its rapid propagation and can be used as a nursery method, contributing to the development of this species as an alternative food and medicinal crop.
- A first approach for the micropropagation of the medicinal halophyte Polygonum maritimum L. and phenolic profile of acclimatized plantsPublication . Custódio, Luísa; Slusarczyk, Sylwester; Matkowski, Adam; Castañeda-Loaiza, Viana; Fernandes, Eliana; Pereira, Catarina; Rodrigues, Maria JoãoPolygonum maritimum L. (sea knotgrass) belongs to a genus commonly used in folk medicine to treat inflammation-related disorders. In vitro pharmacological studies have confirmed these properties that were ascribed to bioactive flavonoids, such as myricetin and quercetin glycosides. Therefore, this study aimed at establishing a micropropagation procedure for sea knotgrass for obtaining standardized materials for its potential commercial cultivation. For that, a complete plant regeneration protocol was developed by improving shoot multiplication from nodal explants, rooting and acclimatization procedures, followed by the assessment of the phenolic profile of the in vitro-produced plants. The combination of 3 mg/L 6-benzylaminopurine (BA) + 0.1 mg/L indole-3-acetic acid (IAA) induced the maximum shoot formation (10.3), which was significantly increased from the first to the second cycle (18.3). The best rooting capacity was observed on shoots derived from the control medium (100%), followed by 2 mg/L kinetin (KIN) (97%) and 3 mg/L BA + 0.1 mg/L IAA (90%); however, the shoot number at the end of the rooting phase was higher on shoots derived from 3 mg/L BA + 0.1 mg/L IAA (6.16). The plant growth regulators used in the multiplication phase influenced survival in the acclimatization process, and plants derived from the control medium had the highest survival percentage (63.1%). Acetone extracts made from aerial organs of micropropagated sea knotgrass showed a predominance of the flavonoid myricetin-3-O-rhamnoside (8.135 mg/g). Overall, the halophyte sea knotgrass was successfully micropropagated showing its potential as a medicinal crop for the extraction of bioactive molecules.
- In vitro cytotoxic effects and apoptosis induction by a methanol leaf extract of carob tree (Ceratonia siliqua L.)Publication . Custódio, Luísa; Escapa, Ana Luísa; Fernandes, Eliana; Fajardo, Alba; Aligué, Rosa; Alberício, Fernando; Neng, Nuno Rosa; Nogueira, José Manuel F.; Romano, AnabelaThis research evaluated the in vitro apoptotic inducing properties of a methanol leaf extract of carob tree (Ceratonia siliqua L.) on a human cervical adenocarcinoma cell line (HeLa). The cell viability effect on a prostate (DU-145), breast (MDA-MB-231) and colon cell line (HCT-166) was also assessed. The effect of the extract on reactive oxygen species (ROS) production by HeLa cells was studied, and a phytochemical evaluation was made by high performance liquid chromatography with diode array detection (HPLC). Total mean yield of identified phenols was 261.1 mg/g DW, and (+)-catechin was the major compound (76.8 mg/g DW). The extract decreased cell viability in a dose-and time-dependent manner, and a more pronounced effect on HeLa line was observed. In vitro cytotoxic activity was associated with apoptosis, but not to the increase of ROS production. Among the tested compounds identified in the extract, the highest activity was detected with gallic acid (GA), (+)-catechin and quercetin, with reductions in HeLa cells viability down to 8.0, 11.9 and 27.1%, respectively. This is the first report on the apoptotic activity of a leaf extract of carob tree on a human cancer cell line, suggesting that it may be a potential source of chemopreventive compounds.