Browsing by Author "Ivanov, Marija"
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- An in vitro study of the origanum minutiflorum O. Schwarz & P. H. Davis and Coriandrum sativum L. essential oils as chronic tonsillitis therapeutics: antibacterial, antibiofilm, antioxidant, and cytotoxic activitiesPublication . Kostić, Marina; Ivanov, Marija; Marković, Tatjana; Sankovic-Babic, Snezana; Barros, Lillian; Calhelha, Ricardo C.; Soković, Marina; Cirić, AnaThe chemical composition and biological activity of essential oils (EOs) from leaves of Origanum minutiflorum O. Schwarz & P.H.Davis, and seeds of Coriandrum sativum L. were investigated. Chemical analyses of EOs were performed and the major components were carvacrol (81.5%) and linalool (69.6%), respectively. The antimicrobial activity was assessed against several bacteria originating from the tonsillar tissue. Activities of EOs against Staphylococcus aureus biofilm were investigated, as well as the effect of the mixture of these EOs and antibiotics against the pathogen. The antioxidant activity of both EOs was determined by TBARS assay, and examined wild oregano EO showed better activity. Also, cytotoxicity of EOs was evaluated in vitro and both EOs showed potential to inhibit further proliferation of tumor cells. This study reported for the first time the effects of EOs on chronic tonsillitis causative pathogens, supporting their role as a natural bioactive therapeutic agent with possible antimicrobial applications.
- Antioxidant extracts of three russula genus species express diverse biological activityPublication . Kostić, Marina; Ivanov, Marija; Fernandes, Ângela; Pinela, José; Calhelha, Ricardo C.; Glamočlija, Jasmina; Barros, Lillian; Ferreira, Isabel C.F.R.; Soković, Marina; Ćirić, AnaThis study explored the biological properties of three wild growing Russula species (R. integra, R. rosea, R. nigricans) from Serbia. Compositional features and antioxidant, antibacterial, antibiofilm, and cytotoxic activities were analyzed. The studied mushroom species were identified as being rich sources of carbohydrates and of low caloric value. Mannitol was the most abundant free sugar and quinic and malic acids the major organic acids detected. The four tocopherol isoforms were found, and polyunsaturated fatty acids were the predominant fat constituents. Regarding phenolic compounds, P-hydroxybenzoic and cinnamic acids were identified in the prepared methanolic and ethanolic extracts, which displayed antioxidant activity through the inhibition of thiobarbituric acid reactive substances (TBARS) formation and oxidative hemolysis; the highest activity was attributed to the R. nigricans ethanolic extract. This is the first report on the antibacterial and antibiofilm potential of the studied species, with the most promising activity observed towards Streptococcus spp. (0.20–0.78 mg/mL as the minimal inhibitory concentration, MIC). The most promising cytotoxic effect was caused by the R. integra methanolic extract on non-small cell lung cancer cells (NCI-H460). Therefore, due to the observed in vitro bioactive properties, the studied mushrooms arise as a source of functional ingredients with potential to be used in novel nutraceutical and drug formulations, which can be used in the treatment of various diseases and health conditions.
- Attalea speciosa mesocarp flour in-depth characterization and its application for the development of new bakery productsPublication . Souza, M.V.S.; Saldanha, Ana Luísa; Pereira, Carla; Ivanov, Marija; Soković, Marina; Steinmacher, Nádia Cristiane; Dias, Maria Inês; Barros, LillianNon-conventional food plants (NFPs) present themselves as a viable and efficient alternative for replacing the food products we consume today. Presenting a huge nutritional, chemical, physical, and biological potential, these plants are usually abundant and not competitors with other vegetable matrices used for human consumption.1 Attalea speciosa (Mart. Ex Spreng - Babassu) is a palm tree from the botanical family Arecaceae found in Brazil, and its mesocarp (BM) represents about 20.4% of the fruit and most of its application it’s in the manufacture of animal feed.2 Thus, the present work aimed to deepen the study the nutritional characterization of BM by AOAC methods; the determination, by chromatographic methods, of free sugars (HPLC-RI), fatty acids (GC-FID), and organic acids content (UPLC-DAD); and the study of the phenolic profile (HPLC-DAD/ESI(MS) and bioactivities (antioxidant, antimicrobial, anti-inflammatory and cytotoxicity activities) of the hydroethanolic extracts. Furthermore, given the possible industrial application, bakery products (bread) were developed with 12, 18, and 24% substitutions of wheat flour by BM flour. BM flour presented low moisture, fat, ash levels, being carbohydrates the main macronutrients, and palmitic (C16:0) and stearic (C18:0) acids were the main fatty acids found in the sample. Regarding phenolic composition, nine phenolic compounds were tentatively identified, six flavan-3-ols (catechin and epicatechin derivatives) and three Oglycosylated flavonoids (quercetin derivatives). Specifically, the flavan-3-ols group represented 99% of total amount of phenolic compounds mainly due to the presence of β-type (Epi)catechin dimer. The hydroethanolic extract of BM showed a high antioxidant capacity to inhibit lipid peroxidation and high anti-hemolytic capacity, showing an IC50 99% and 75% more effective than the positive control used (Trolox) in these assays, respectively. Moreover, it was able to inhibit all the tumor cell lines tested (MCF-7, NCIH460, CaCo, and AGS), however, it showed some toxicity towards healthy cells of PLP2 and VERO lines. The results obtained for the antibacterial activity of the BM extract were lower when compared to the positive controls used (E211 and E224). Comparing the results obtained with 100% wheat flour products, it was noted that the formulations with substitutions by BM presented remarkable results, providing a reasonable increase in the PUFAs content (despite decreasing protein content), maintaining the antioxidant capacity with lower IC50 values than the positive control used, and not presenting hepatotoxic activity (first validation of this flour for the incorporation in food matrices). In general, the formulation B24 was the one that presented greater similarity to the control bread (B0), relatively to the nutritional aspects, however, concerning the physical parameters it was the one that presented bigger discrepancy, especially regarding the specific volume, texture, and the color parameter L* (luminosity), tending for a darker and opaque coloration. All the formulations presented higher percentages of loss of rheological characteristics in the first three days of elaboration, however, formulation B24 was the one that presented the lowest percentage. This study presented innovative results regarding the nutritional, chemical and bioactive characterization of both the babassu mesocarp itself and the bread made from it, showing great potential to be applied in the food industry, however, there is still much to explore, especially in preparations that do not yet use non-conventional ingredients.
- Bioactive compounds and functional properties of herbal preparations of Cystus creticus L. collected from Rhodes IslandPublication . Mocan, Andrei; Fernandes, Ângela; Calhelha, Ricardo C.; Gavrilas, Laura; Ferreira, Isabel C.F.R.; Ivanov, Marija; Barros, Lillian; Babotă, MihaiThe members of Cystus genus are perenial shrubs with a well-established use in traditional medicine. Among these, C. creticus is the most popular, herbal preparations obtained from its aerial parts being recognized as antimicrobial, antitumor and anti-inflammatory agents. The present study aimed to evaluate phytochemical profile and bioactive potential of aqueous and hydroethanolic extracts of C. creticus aerial parts harvested from two different areas of Rhodes island. LC-DAD-ESI/MSn analysis revealed the presence of myricetin and quercetin glycosides as main compounds, especially in aqueous extracts, being probably responsible for their enhanced antioxidant and antimicrobial potential. On the other side, hydroethanolic preparations exerted a strong anti-inflammatory and anti-biofilm activity. Our findings suggest that the use of solvents with intermediate polarity can assure the best recovery of bioactive compounds from C. creticus, increasing the extraction yield for other non-phenolic compounds which can enhance therapeutic potential of the extract through a synergistic action.
- Bioactive potential of Tripleurospermum inodorum with detailed insight into anti-inflammatory activity through in vitro, in vivo evaluations and network pharmacologyPublication . Ivanov, Marija; Aleksandrov, Aleksandra Popov; Božunović, Jelena; Dias, Maria Inês; Calhelha, Ricardo C.; Kulaš, Jelena; Barros, Lillian; Ferreira, Isabel C.F.R.; Stojković, DejanThis study evaluated Tripleurospermum inodorum extract for cytotoxic, anti-inflammatory, antioxidant, antimicrobial, and antibiofilm properties, alongside its phenolic profile, predicted pharmacological interactions and in vivo anti-inflammatory properties. The methanolic extract of T. inodorum was rich in apigenin derivatives, including apigenin-O-pentoside (5.234 mg/g) and apigenin-O-acetyl hexoside (4.929 mg/g), as identified using LC-DAD-ESI/MSⁿ. The extract demonstrated potent anti-inflammatory activity (IC₅₀ = 8.4 μg/mL) by inhibiting nitric oxide production in a RAW 264.7 macrophage model, a key mechanism in controlling inflammatory responses. Its cytotoxicity against NCI-H460 lung carcinoma cells (GI₅₀ = 62.9 μg/mL) suggests potential for targeting inflammation-driven carcinogenesis. Antioxidant activity, ranging from 204.4 (FRAP) to 442.2 (ABTS) mmol of gallic acid equivalents per 100 mg dry weight, highlights its role in mitigating oxidative stress—a critical driver of chronic inflammation. The extract also displayed moderate antimicrobial activity (MIC: 3–12 mg/mL) and strong antibiofilm potential (> 70% inhibition in a crystal violet assay), which are essential for managing infection-associated inflammation. Network pharmacology revealed that dominant phenolic compound act as aldose reductase inhibitors, targeting inflammatory pathways linked to metabolic stress. In vivo assessment using a xylene-induced ear edema model revealed a dose-dependent, biphasic anti-inflammatory effect, with lower doses (125 and 250 mg/kg) exhibiting greater efficacy compared to the highest dose (500 mg/kg), suggesting a hormetic response that emphasizes the importance of optimal dosing. These findings indicate that the methanolic extract of T. inodorum possesses a broad spectrum of bioactivities relevant to inflammation control and supports its further development as a source of novel anti-inflammatory therapeutics.
- Biochemical and molecular profiling of wild edible mushrooms from huila, AngolaPublication . Kissanga, Raquel; Liberal, Ângela; Diniz, Ines; Rodrigues, Ana S. B.; Baptista-Ferreira, Joao L.; Batista, Dora; Ivanov, Marija; Soković, Marina; Ferreira, Isabel C.F.R.; Fernandes, Ângela; Barros, Lillian; Catarino, LuísThe harvesting, processing, and sale of wild edible mushrooms (WEM) is a relevant economic activity in Angola and a good example of the use of non-wood forest products for food. Although there is deep traditional knowledge about the general properties of WEMs, a huge gap remains in detailed scientific knowledge. Thus, this study aimed to investigate the socio-economic importance of the species sold at local markets in Huila, Angola, from their molecular identification to the assessment of their nutritional, chemical, and bioactive profiles. From the eight WEM morphotypes studied, five were identified based on phenotypical and molecular approaches (four Russula spp., and Amanita loosei). The studied mushrooms proved to be a rich source of carbohydrates, proteins, and ashes, also presenting low amounts of fat. Chemical analyses further revealed mannitol as the main free sugar in all samples, and organic acids, namely, oxalic, quinic, malic, citric, and fumaric acids in low amounts. Additionally, the alpha-tocopherol isoform and monounsaturated fatty acids were predominant. Regarding phenolic acids, protocatechuic, p-hydroxybenzoic, p-coumaric, and cinnamic acids were detected in all mushroom hydroethanolic extracts, being responsible for their antioxidant, antibacterial, and antifungal activities. Our investigation contributes to the identification and knowledge of WEMs as important complementary food sources in Angola, some of which were reported for the first time, promoting their utilization as a basis of nutritional and functional ingredients, as being able to be part of a balanced diet and to be used in new bio-based formulations.
- Camphor and eucalyptol—anticandidal spectrum, antivirulence effect, efflux pumps interference and cytotoxicityPublication . Ivanov, Marija; Kannan, Abhilash; Stojković, Dejan; Glamočlija, Jasmina; Calhelha, Ricardo C.; Ferreira, Isabel C.F.R.; Sanglard, Dominique; Soković, MarinaCandida albicans represents one of the most common fungal pathogens. Due to its increasing incidence and the poor efficacy of available antifungals, finding novel antifungal molecules is of great importance. Camphor and eucalyptol are bioactive terpenoid plant constituents and their antifungal properties have been explored previously. In this study, we examined their ability to inhibit the growth of different Candida species in suspension and biofilm, to block hyphal transition along with their impact on genes encoding for efflux pumps (CDR1 and CDR2), ergosterol biosynthesis (ERG11), and cytotoxicity to primary liver cells. Camphor showed excellent antifungal activity with a minimal inhibitory concentration of 0.125–0.35 mg/mL while eucalyptol was active in the range of 2–23 mg/mL. The results showed camphor’s potential to reduce fungal virulence traits, that is, biofilm establishment and hyphae formation. On the other hand, camphor and eucalyptol treatments upregulated CDR1; CDR2 was positively regulated after eucalyptol application while camphor downregulated it. Neither had an impact on ERG11 expression. The beneficial antifungal activities of camphor were achieved with an amount that was non-toxic to porcine liver cells, making it a promising antifungal compound for future development. The antifungal concentration of eucalyptol caused cytotoxic effects and increased expression of efflux pump genes, which suggests that it is an unsuitable antifungal candidate.
- Caracterização fenólica e nutricional das infusões de cinco plantas aromáticas e suas propriedades bioativasPublication . Caleja, Cristina; Finimundy, Tiane C.; Pereira, Carla; Barros, Lillian; Calhelha, Ricardo C.; Soković, Marina; Ivanov, Marija; Carvalho, Ana Maria; Rosa, Eduardo; Ferreira, Isabel C.F.R.As infusões de plantas são aplicadas há séculos na medicina popular para fins medicinais e nutricionais [1]. Devido à sua importância económica e elevado consumo, tem-se verificado um crescente interesse não só pela sua composição química como também pelas suas diversas propriedades bioativas [2]. Neste estudo, foram selecionadas cinco espécies de plantas aromáticas: Erica australis L., Genista tridentata L., Melissa officinalis, L., Mentha spicata L. e Prunella vulgaris L. Foram preparadas infusões de cada planta e analisadas quanto à sua composição fenólica (HPLC/DAD-ESI/MS) e propriedades bioativas. Para a atividade antioxidante foram utilizados métodos que avaliam a inibição da pero xidação lipídica (TBARS) e da hemólise oxidativa (OxHLIA). A atividade antimicrobiana foi avaliada pelo método de microdiluição em placa utilizando bactérias Gram + (Bacillus cereus e Listeria monocytogenes) e Gram - (Escherichia coli e Salmonela typhimurium) e fungos (Aspergillus niger, Aspergillus versicolor, Penicillium funiculosum e Penicillium verrucosum). A atividade citotóxica foi testada usando o método colorimétrico da sulforrodamina B em diferentes linhas celulares tumorais MCF-7 (adenocarcinoma da mama), NCI-H460 (carcinoma de pulmão), HeLa (carcinoma cervical), HepG2 (carcinoma hepatocelular) e numa cultura primária de células não tumorais (PLP2); para a atividade anti-inflamatória utilizou-se a linha de macrófagos RAW 246.7. Foram identificados 52 compostos fenólicos, sendo que o composto maioritário nas infusões de M. officinalis, M. spicata e P. vulgaris foi o ácido rosmarínico, enquanto na de E. australis os mais abundantes foram os derivados de flavonoides identificados como quercetina e genisteína. As infusões de P. vulgaris e M. officinalis apresentaram os melhores resultados nos ensaios TBARS (IC50 de 4,2±0,04 μg/mL) e OxHLIA (IC50 de 24,8±0,3 μg/mL), respetivamente. Nenhuma das infusões mostrou hepatotoxicidade em células não-tumorais (GI50>400 μg/mL). A infusão de M. spicata apresentou os melhores resultados de citotoxicidade contra a linha celular MCF-7 (GI50 de 283±10 μg/mL) e de atividade antimicrobiana, em especial contra a Salmonela typhimurium (MIC de 0,25 mg/mL) e os fungos Aspergillus versicolor (MIC de 0,25 mg/mL) e Penicillium verrucosum (MIC de 0,25 mg/mL), sendo a única planta a apresentar atividade anti-inflamatória (GI50 de 324±5 μg/mL). Este trabalho demonstrou o potencial das infusões destas plantas, ricas em compostos fenólicos com propriedades bioativas, para futura aplicação em diferentes sectores industriais.
- Caracterización detallada de la harina del mesocarpio de Attalea speciosa y su aplicación para el desarrollo de nuevos productos de panaderíaPublication . Souza, M.V.S.; Saldanha, Ana Luísa; Pereira, Carla; Ivanov, Marija; Soković, Marina; Steinmacher, Nádia Cristiane; Dias, Maria Inês; Barros, Lillian; Morais, CarlosLas plantas alimenticias no convencionales se presentan como una alternativa viable y eficiente para reemplazar los productos alimenticios que consumimos hoy. Presentando un enorme potencial nutricional, químico, físico y biológico, estas plantas suelen ser abundantes y no competidoras de otras matrices vegetales [1]. Attalea speciosa (Mart. Ex Spreng - Babassu) es una palmera que se encuentra en Brasil, y su mesocarpio (MB) representa cerca del 20,4% del fruto, presentando aplicabilidad en la fabricación de alimentos para animales [2]. Así, el presente trabajo tuvo como objetivo profundizar el estudio de la caracterización nutricional de MB por métodos de la AOAC; la determinación del contenido de azúcares libres (HPLC-RI), ácidos grasos (GC-FID), ácidos orgánicos (UPLC-DAD) y compuestos fenólicos (HPLCDAD/ ESI(MS); y propiedades bioactivas (antioxidante, antimicrobiana, antiinflamatoria, citotóxica y nefrotoxicidad) de los extractos hidroetanólicos. Además, se desarrollaron productos de panadería (pan) con 12, 18 y 24% de sustitución de harina de trigo por harina MB. La harina presentó bajos niveles de humedad, grasa y cenizas, siendo los carbohidratos los principales macronutrientes, y los ácidos palmítico y esteárico los principales ácidos grasos. Se identificó tentativamente nueve compuestos fenólicos, seis flavan-3-oles (99% de la cantidad total, sobre todo por la presencia del dímero de (epi)catequina tipo β) y tres flavonoides O-glicosilados. El extracto hidroetanólico mostró una alta capacidad antioxidante y antihemolítica, mostrando una IC50 de 99% y 75% más efectiva que el control positivo (Trolox), respectivamente. Además, fue capaz de inhibir todas las líneas celulares tumorales, per mostró cierta toxicidad hacia las células sanas. Para la actividad antibacteriana fueron obtenidas concentraciones mínimas inhibitorias inferiores a la de los controles positivos (E211 y E224). Finalmente, se observó que las formulaciones de pan con sustituciones presentaron un aumento razonable en el contenido de PUFAs, manteniendo la capacidad antioxidante con valores de IC50 más bajos que los del control positivo, y no presentando actividad hepatotóxica (primera validación de esta harina para la incorporación en matrices alimentarias). En general, la formulación B24 fue la que presentó mayor similitud con el pan control, en relación a los aspectos nutricionales, sin embargo, en cuanto a los parámetros físicos fue la que presentó mayor discrepancia (en volumen específico, textura, y luminosidad). Todas las formulaciones presentaron mayores porcentajes de pérdida de características reológicas en los primeros tres días de elaboración. Este estudio presentó resultados innovadores en cuanto a la caracterización nutricional, química y bioactiva tanto del propio mesocarpio de babasú como del pan elaborado a partir de él, mostrando un gran potencial para ser aplicado en la industria alimentaria, sin embargo, aún queda mucho por explorar, especialmente en preparaciones que todavía no use ingredientes no convencionales
- Challenges of traditional herbal teas: Plant infusions and their mixtures with bioactive propertiesPublication . Caleja, Cristina; Finimundy, Tiane C.; Pereira, Carla; Barros, Lillian; Calhelha, Ricardo C.; Soković, Marina; Ivanov, Marija; Carvalho, Ana Maria; Rosa, Eduardo; Ferreira, Isabel C.F.R.In order to characterize and study the bioactivities of individual plant species and to determine how these characteristics are modified when preparing blends, five different plant species were selected: Erica australis L., Genista tridentata L., Melissa officinalis L., Mentha spicata L., and Prunella vulgaris L. Infusions prepared from each plant species and from three selected mixtures were analyzed in terms of nutritional values, phenolic compositions, and bioactive properties (antioxidant, antimicrobial, cytotoxic, and anti-inflammatory activities). The major compound detected in M. officinalis, M. spicata, and P. vulgaris infusions was rosmarinic acid, whilst in E. australis and G. tridentata flavonoid derivatives such as quercetin and genistein were identified. P. vulgaris and M. officinalis presented the best results in TBARS and OxHLIA assays, respectively. M. spicata and all mixtures presented anti-inflammatory activity. M. spicata showed the best cytotoxic properties and antimicrobial activity, and none of the infusions showed hepatotoxicity for non-tumour cells.