Caracterização química e avaliação das propriedades bioativas das sumidades floridas de Calluna vulgaris (L.) Hull

Nos últimos tempos tem-se observado um aumento do interesse da população no sentido de melhorar o estilo e a qualidade de vida, dando-se cada vez mais ênfase à importância dos benefícios decorrentes do consumo dos produtos de origem natural. Dado este interesse, e também como resultado da imensa variedade química e biológica associada a estes produtos, a ciência tem aumentado o foco neste tipo de matrizes naturais, sendo cada vez mais importante e de reconhecimento global, a exploração destes recursos. A Calluna vulgaris (L.) Hull, vulgarmente conhecida por urze, é um arbusto que se encontra intensamente presente nas paisagens de várias regiões e é tradicionalmente utilizado no tratamento de diferentes patologias. Os estudos descritos na literatura demonstram o seu elevado conteúdo em compostos fenólicos, bem como o seu promissor potencial biológico. Na presente dissertação as sumidades floridas de C. vulgaris foram caracterizadas relativamente ao seu valor nutricional, perfil químico e propriedades bioativas. Assim, numa primeira fase, procedeu-se à sua caracterização nutricional através da determinação do teor em cinzas, gorduras, proteínas, hidratos de carbono e do seu valor energético utilizando métodos oficiais de análise de alimentos (AOAC); foi também determinado o teor em ácidos gordos (GC-FID), ácidos orgânicos (UFLC-PDA), açúcares livres (HPLC-RI) e tocoferóis (HPLC-fluorescência). Através da extração sucessiva por maceração à temperatura ambiente e sob vigorosa agitação, foram obtidos cinco extratos distintos, usando solventes orgânicos de polaridade crescente (n-hexano, diclorometano, acetato de etilo, acetona e metanol); foram também obtidos dois extratos aquosos por infusão e decocção, formas tradicionais de utilização da urze. O perfil fenólico de cada um destes extratos foi avaliado por HPLC-DAD-ESI/MS. Os cinco extratos orgânicos e dois aquosos foram analisados relativamente à sua bioatividade (atividade antioxidante, citotóxica, anti-inflamatória e antibacteriana). O potencial antioxidante foi avaliado através de dois métodos distintos: inibição da peroxidação lipídica em tecidos cerebrais de porco (TBARS) e inibição da hemólise oxidativa (OxHLIA). As propriedades citotóxicas dos sete extratos foram estudadas em cinco linhas celulares tumorais humanas: AGS (adenocarcinoma gástrico), HeLa (adenocarcinoma cervical), HepG2 (carcinoma hepatocelular), MCF-7 (adenocarcinoma de mama) e NCI-H460 (carcinoma de pulmão); por sua vez, a citotoxicidade em células não tumorais foi avaliada numa cultura de células não tumorais: PLP2 (cultura primária de células de fígado de porco). A atividade anti-inflamatória foi estudada utilizando uma linha celular de macrófagos de rato RAW 264.7. A atividade antibacteriana foi avaliada através da técnica de microdiluição, juntamente com o método colorimétrico de deteção rápida com cloreto de p-iodonitrotetrazólio (INT). Por último, e como resultado do potencial antibacteriano demonstrado, os extratos de metanol, acetona, infusão e decocção foram analisados no sentido de verificar o seu efeito em bactérias que fazem parte da microbiota vaginal. Na análise da composição nutricional verificou-se que os hidratos de carbono são os macronutrientes maioritários, seguidos das proteínas, lípidos e cinzas. Os açúcares identificados foram a glucose e a frutose. Verificou-se também a presença de quatro ácidos orgânicos (ácido oxálico, quínico, ascórbico e cítrico), das quatro isoformas de tocoferóis e, finalmente, foram identificados 16 ácidos gordos. Através da análise do perfil fenólico dos extratos das sumidades floridas da urze foram identificados 12 compostos fenólicos (2 ácidos fenólicos e 10 flavonoides). A miricetina 3-O-glucosido e a miricetina-O-ramnósido foram as moléculas maioritárias em todos os extratos, com exceção do extrato de diclorometano que não apresentou nenhum dos compostos fenólicos identificados. De uma forma geral, o teor de compostos fenólicos aumenta com o aumento do grau de polaridade do solvente de extração. No que respeita às bioatividades, os resultados apresentam diferenças consoante o tipo de extrato: enquanto para a atividade antioxidante o extrato que se destacou foi o de acetona, para a citotoxicidade foi o de acetato de etilo, para a atividade anti-inflamatória o de n-hexano e para a atividade antibacteriana (bactérias patogénicas e pertencentes à microbiota urogenital) destacaram-se os extratos de acetona e metanol. Como resultado do elevado potencial antibacteriano demonstrado pelo extrato de acetona, tanto para as bactérias patogénicas como para os Lactobacillus pertencentes à microbiota urogenital, este foi fracionado por cromatografia em coluna de sílica gel através de um sistema de eluentes de polaridade crescente. As 10 frações obtidas foram caracterizadas relativamente ao seu perfil fenólico e às propriedades antibacterianas. A análise do perfil fenólico das 10 frações resultou na identificação de 47 compostos fenólicos (1 ácido fenólico, 16 flavan-3-óis, 2 flavanonas e 24 flavanóis), dos quais os dímeros do tipo B da (epi)catequina, (-)-epicatequina e (+)-catequina revelaram ser os compostos maioritários. Apesar de as frações não serem eficazes para todas as bactérias patogénicas e não serem todas beneficiadas com o fracionamento, revelaram eficácia contra alguns destes microrganismos sem afetar os Lactobacillus da microbiota vaginal, destacando-se as frações 7 e 8. O presente trabalho permitiu comprovar o potencial biológico associado a esta espécie, em particular e para algumas concentrações o seu potencial antibacteriano contra bactérias patogénicas e a capacidade de preservar a microbiota vaginal. Além disso, também se demonstrou que as formas tradicionais de consumo da mesma são as que revelam maior atividade antibacteriana, servindo de base científica para alertar o consumidor dos potenciais associados à utilização deste arbusto.En los últimos años se ha observado un incremento del interés y atención de la población en mejorar el estilo y la calidad de vida, dando cada vez más énfasis a la importancia de los beneficios derivados del consumo de productos de origen natural. Debido a este interés, y también como resultado de la inmensa variedad química y biológica asociada a estos productos, la ciencia y la industria ha unificado fuerzas y enfocado su interés en la explotación de estos recursos. La Calluna vulgaris (L.) Hull, vulgarmente conocida por brezo, es un arbusto que se encuentra intensamente presente en los paisajes de varias regiones y es tradicionalmente utilizada en el tratamiento de diferentes patologías. Los estudios descritos en la literatura científica demuestran su alto contenido en compuestos fenólicos, así como su prometedor potencial biológico asociado a esta especie. En la presente disertación, las sumidades floridas de C. vulgaris, se caracterizaron por su valor nutricional, su perfil químico y potencial bioactiva. Así, en una primera fase, se procedió a su caracterización nutricional mediante la determinación del contenido en cenizas, grasas, proteínas, hidratos de carbono y su valor energético utilizando métodos oficiales de análisis de comida (AOAC); se determinó el contenido en ácidos grasos (GC-FID), los ácidos orgánicos (UFLC-PDA), los azúcares libres (HPLC-RI) y los tocoferoles (HPLC-fluorescencia). A través de la extracción sucesiva por maceración, a la temperatura ambiente y bajo una vigorosa agitación, se obtuvieron cinco extractos distintos, usando disolventes orgánicos de polaridad creciente (n-hexano, diclorometano, acetato de eilo, acetona y metanol); se obtuvieron dos extractos acuosos por infusión y decocción, formas tradicionales de utilización del brezo. El perfil fenólico de cada uno de estos extractos fue evaluado por HPLC-DAD-ESI/MS. Los cinco extractos orgánicos y dos acuosos fueron evaluados en relación a su bioactividad (actividad antioxidante, citotóxica, anti-inflamatoria y antibacteriana). El potencial antioxidante fue evaluado a través de dos métodos distintos: inhibición de la peroxidación lipídica en tejidos cerebrales de cerdo (TBARS) y de la inhibición del hemólisis oxidativa (OxHLIA). Las propiedades citotóxicas de los distintos extractos se estudiaron en cinco líneas celulares tumorales humanas: AGS (adenocarcinoma gástrico), HeLa (adenocarcinoma cervical), HepG2 (carcinoma hepatocelular), MCF-7 (adenocarcinoma de mama) y NCI-H460 (carcinoma de pulmón); a su vez, la citotoxicidad en células no tumorales se evaluó en una cultura de células no tumorales: PLP2 (cultivo primario de células de hígado de cerdo). La actividad anti-inflamatoria fue estudiada en una línea celular de macrófagos de rata RAW 264.7. La actividad antibacteriana fue evaluada a través de la técnica de microdilución junto con el método colorimétrico de detección rápida con cloruro de p-yodonitrotetrazolio (INT). Por último, y como resultado del potencial antibacteriano demostrado, los extractos de metanol, acetona, infusión y decocción se evaluaron para comprobar su efecto en las bacterias que forman parte de la microbiota vaginal. Con la evaluación de la composición nutricional, se verificó que los hidratos de carbono son los macronutrientes mayoritarios, seguido de las proteínas, lípidos e cenizas. Los azúcares identificados fueron la glucosa y frutosa. Se verificó también la presencia de cuatro ácidos orgánicos (ácido oxálico, quínico, ascórbico y cítrico), de los cuatro isoformas de tocoferoles y, finalmente, de 16 ácidos grasos. El análisis del perfil fenólico de los extractos, fueron identificados 12 compuestos fenólicos (2 ácidos fenólicos y 10 flavonoides). La miricetina 3-O-glucosido y la miricetina-O-ramnósido fueron los compuestos presentes en mayor cantidad en todos los extractos, excepto en el extracto de diclorometano que no ha presentado ninguno de los compuestos fenólicos identificados. De forma general, el contenido en compuestos fenólicos es mayor cuanto mayor es el grao de polaridad del solvente de extracción. En cuanto a las bioactividades, los resultados presentan diferencias según el tipo de extracto: mientras que para la actividad antioxidante el extracto destacado fue el de acetona, para la citotoxicidad fue el de acetato de etilo, para la actividad anti-inflamatoria el de n-hexano y para la actividad antibacteriana (bacterias patógenas y pertenecientes a la microbiota urogenital) los extractos de acetona y metanol. Como resultado del elevado potencial antibacteriano demostrado por el extracto de acetona, tanto para las bacterias patógenas como para los Lactobacillus pertenecientes a la microbiota urogenital, el extracto fue fraccionado por cromatografía en columna de sílice gel a través de un sistema de eluentes de polaridad creciente. Las 10 fracciones obtenidas se fueron caracterizadas por su perfil fenólico y sus propiedades antibacterianas. El análisis del perfil fenólico de las 10 fracciones resultó en la identificación de 47 compuestos fenólicos (1 ácido fenólico, 16 flavan-3-ois, 2 flavanonas y 24 flavanoles), de los cuales los dímeros del tipo B de la (epi)catequina, (-)-epicatequina y (+)-epicatequina revelaron mayor concentración. Aunque las fracciones utilizadas no han sido eficaces para todas las bacterias patógenas, revelaron eficacia contra algunos de los microorganismos patógenos, sin afectar a los Lactobacillus de la microbiota vaginal, destacándose las fracciones 7 y 8. El presente trabajo permitió comprobar el potencial biológico asociado a esta especie, en particular y para algunas concentraciones, el potencial antibacteriano contra bacterias patógenas y la capacidad de preservar la microbiota vaginal. Además, también se ha demostrado que las formas tradicionales de consumo de la misma son las que revelan una mayor actividad antibacteriana, sirviendo de base científica para alertar al consumidor de los potenciales asociados a la utilización de este arbusto

Chemical composition of cynara cardunculus L. var. altilis heads: the impact of harvesting time

Cardoon is a multi-purpose crop with several industrial applications, while the heads (capitula) are edible and commonly used in various dishes of the Mediterranean diet. Several reports in the literature study the chemical composition of the various plants parts (leaves, flower stalks, bracts, seeds) aiming to industrial applications of crop bio-waste, whereas for the heads, most of the studies are limited to the chemical composition and bioactive properties at the edible stage. In the present study, cardoon heads were collected at six different maturation stages and their chemical composition was evaluated in order to determine the effect of harvesting stage and examine the potential of alternative uses in the food and nutraceutical industries. Lipidic fraction and the content in fatty acids, tocopherols, organic acids, and free sugars were determined. Lipidic content decreases with the maturation process, while 22 fatty acids were detected in total, with palmitic, oleic, and linoleic acids being those with the highest abundance depending on harvesting time. In particular, immature heads have a higher abundance in saturated fatty acids (SFA), whereas the samples of mature heads were the richest in monounsaturated fatty acids (MUFA). The a-tocopherol was the only isoform detected being present in higher amounts in sample Car B (619 μ g/100 g dw). Oxalic, quinic, malic, citric and fumaric acids were the detected organic acids, and the higher content was observed in sample Car E (15.7 g/100 g dw). The detected sugars were fructose, glucose, sucrose, trehalose and raffinose, while the highest content (7.4 g/100 g dw) was recorded in sample Car C. In conclusion, the maturation stage of cardoon heads influences their chemical composition and harvesting time could be a useful means to increase the quality and the added value of the final product by introducing this material in the food and nutraceutical industries.This work was financially support by national funds FCT/MCTES to CIMO (UIDB/00690/2020). We also thank the FCT for the PhD grant (SFRH/BD/146614/2019) of F. Mandim. The authors are also grateful to the financial support though the TRANSCoLAB (0612_TRANS_CO_LAB_2_P).info:eu-repo/semantics/publishedVersio

Combined effect of biostimulants and mineral fertilizers on crop performance and fruit quality of watermelon plants

The aim of the present study was to evaluate the possible effects of two different biostimulant formulations at different application regimes and combined or not with mineral fertilizers (e.g., W1–W8, including the control treatment (no formulations added)) on the yield parameters and fruit quality of watermelon plants. The highest yield was recorded for the W5 treatment due to the formation of more fruit. The highest content of fat, proteins and ash was recorded for treatment W1, whereas carbohydrates were the most abundant in the control treatment, resulting also in the highest energetic value. The main detected sugars in all the tested samples were sucrose and fructose, which were the highest for the W4 and W5 treatments (sucrose) and W4 treatment (fructose). Malic and citric acid were the most abundant compounds, especially in the W4 treatment. In terms of tocopherols, only -tocopherol was detected, with the highest amounts being recorded for the W4 treatment. Regarding bioactive properties, the lowest IC50 values for OxHLIA were recorded for the W2, W3 and W8 formulations. Moreover, all the extracts exhibited significant anti-inflammatory activity comparable to the positive control, while a variable efficacy of the tested extracts against the studied bacteria and fungi was recorded. In conclusion, our results indicate that simple agronomic practices such as biostimulant application may improve crop performance and improve the proximal composition and the overall quality of watermelon fruit within the context of sustainable crop production.This research was funded by Agrology S.A., grant number 6089. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to the CIMO (UIDB/00690/2020 and UIDP/00690/2020) and SusTEC (LA/P/0007/2020). In addition, national funding was received by F.C.T., P.I., through the institutional scientific employment program contract for Â.F. This work is funded by the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project GreenHealth (Norte-01-0145-FEDER-000042).info:eu-repo/semantics/publishedVersio

How does the maturation state of cardoon bracts influence its phenolic composition and bioactivity?

Cynara cardunculus L. (cardoon) is an herbaceous plant native to the Mediterranean basin. It is widely used due to its nutritional, pharmacological, and industrial applications [1]. Besides its consumption in various recipes, due to its rich nutritional composition and health-promoting effects, it is also widely used in several industry sectors (e.g., cheese manufacturing, and in the production of pharmaceuticals, bioenergy, and biomass). Its multiple applications are extremely important for the enhancement of the species‘ added value [2]. However, factors such as genetic information, plant tissue, and maturity state can influence the potential of the species [1,2]. The study of the influence of all these variables is extremely important for its proper use, as well for its economic valorisation. Cardoon bracts were collected in Greece at eight harvesting dates (Principal Growth Stage between 5 and 8/9). The phenolic composition of their hydroethanolic extracts was analysed by HPLC-DAD-ESI/MS. The antioxidant activity was evaluated with two cell-based assays: thiobarbituric acid reactive substances (TBARS) formation inhibition and oxidative haemolysis assay (OxHLIA). The anti-inflammatory activity was evaluated by determining the extracts‘ ability to inhibit nitric oxide production in a murine macrophage cell line. Finally, cytotoxicity was assessed against four human tumour cell lines and a porcine liver primary cell culture using the sulforhodamine B assay. Twelve phenolic compounds were tentatively identified, with 3,5-O-dicaffeoylquinic acid and apigenin-7-O-glucuronide being the major compounds detected. Immature bracts presented the highest phenolic compounds content, as also the highest anti-inflammatory and cytotoxic activities. Regarding the antioxidant activity, immature bracts revealed the highest ability to inhibit the formation of TBARS. For OxHLIA, cardoon bracts harvested at more advanced maturation stages were the ones that revealed the highest activity. In conclusion, it was verified that the maturation stage influences the phenolic composition and the biological potential of cardoon bracts. However, more studies are needed to understand and correlate the compounds responsible with the observed bioactivities.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020); to the national funding by FCT, P.I., through the institutional scientific employment program-contract for M.I. Dias and L. Barros, and individual scientific employment program-contract to J. Pinela (CEECIND/01011/2018); for F. Mandim PhD grant (SFRH/BD/146614/2019); to FEDER through the Regional Operational Program North 2020, under the Project GreenHealth (Norte-01-0145-FEDER-000042).info:eu-repo/semantics/publishedVersio

Exploring the impacts of sorghum (Sorghum bicolor L. Moench) germination on the flour’s nutritional, chemical, bioactive, and technological properties

Germination is a natural, simple, and economical process used to improve the quality of nutritional and technological grains. In this study, native and sprouted sorghum flours were characterized regarding their technological properties (particle size distribution, water, and oil absorption capacity, swelling power and solubility, microscopy of starch granules, and pasting and thermal properties). Nutritional and phytochemical characterization profiles, including free sugars, fatty acids, organic acids, tocopherols, and phenolic compounds, were explored through chromatographic methods. The antioxidant, anti-inflammatory, and cytotoxic activities of the respective hydroethanolic extracts were also evaluated. The results showed that the germination process caused significant changes in the flour composition and properties, causing reduced gelatinization temperature and retarded starch retrogradation; an increased content of free sugars and total organic acids; and a decreased content of tocopherols and phenolic compounds. In terms of bioactivity, the sprouted sorghum flour extract showed better lipid-peroxidation-inhibition capacity and none of the extracts revealed hepatotoxicity or nephrotoxicity, which are important results for the validation of the use of the flours for food purposes. Germination is an efficient and alternative method for grain modification that gives improved technological properties without chemical modification or genetic engineering.This research was funded by the Foundation for Science and Technology (FCT), P.I. (FCT, Portugal) through national funds FCT/MCTES to CIMO (UIDB/00690/2020 and UIDP/00690/2020), SusTEC (LA/P/0007/2020), and UIDB/50006/2020. National funding was provided by FCT, P.I., through the institutional scientific employment program contract for M.I.D. (10.54499/CEECINST/00016/2018/CP1505/CT0004) and C.P. (10.54499/CEECINST/00016/2018/CP1505/CT0010), and funding for B.H.P and F.M. came from grants (2023.02731.BD and SFRH/BD/146614/2019, respectively).info:eu-repo/semantics/publishedVersio

Synthesis, photochemical and in vitro cytotoxic evaluation of new iodinated aminosquaraines as potential sensitizers for photodynamic therapy

In this work, several benzothiazole-based aminosquaraine dyes, displaying strong absorption within the so-called phototherapeutic window (650–800 nm), were synthesized. The ability, of all the new dyes, to generate singlet oxygen was assessed by determining the correspondent phosphorescence emission and through the comparison with a standard. The quantum yields of singlet oxygen generation were determined and exhibited to be strongly dependent on the nature of the amino substituents introduced in the squaric ring. The photodynamic activity of the synthesized dyes was tested against four human tumor cell lines: breast (MCF-7), lung (NCI-H460), cervical (HeLa) and hepatocellular (HepG2) carcinomas; and a non-tumor porcine liver primary cell culture (PLP2). All the compounds synthesized were found to be able to inhibit tumor cells growth upon irradiation more than in the dark, in most of the cases, very significantly. Considering the photodynamic activity exhibited and the low toxicity displayed for the non-tumor cells, some of the synthetized dyes can be regarded as potential candidates as photosensitizers for PDT.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER for CIMO (UID/AGR/00690/2013) and CQ-VR (UID/QUI/00616/2013) financial support, as also for R. Calhelha contract.info:eu-repo/semantics/publishedVersio

Phenolic profile and effects of acetone fractions obtained from the inflorescences of: Calluna vulgaris (L.) Hull on vaginal pathogenic and non-pathogenic bacteria

The phenolic profile and the antibacterial activity against pathogenic commensal vaginal bacteria exhibited by different fractions of the acetone extract of heather was assessed. The acetone extract of Calluna vulgaris (L.) Hull was fractionated by silica gel column chromatography through an eluent system of increasing polarity, obtaining 10 different fractions (Fr 1 to Fr 10). The phenolic profile was analyzed by HPLCDAD- ESI/MS. Type B (epi)catechin dimers, (-)-epicatechin and (+)-catechin were the main phenolic compounds present in the fractions. The antibacterial activity was also analyzed against pathogenic bacteria and the effect in the beneficial microflora was also accessed. Some of the obtained fractions revealed the capacity to inhibit pathogenic microorganisms without affecting the beneficial microflora, especially Fr 7 and Fr 8. For instance, Neisseria gonorrhoeae was inhibited by both of the fractions, while Fr 7 was more active against Klebsiella pneumoniae and Morganella morganii, and Fr 8 against methicillin resistance Staphylococcus aureus (MRSA) and methicillin susceptible Staphylococcus aureus (MSSA), without affecting Lactobacillus strains. This study corroborates the therapeutic use of this matrix in traditional medicine.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programmer PT2020 for financial support to CIMO (UID/AGR/00690/2019), CQ-VR (UID/QUI/00616/2013), and to FEDER-Interreg España-Portugal program for financial support through the project 0377_Iberphenol_6_E. The authors are also grateful to the Foundation for Science and Technology (FCT, Portugal) for Sandrina A. Heleno (SFRH/BPD/101413/2014) grant and L. Barros contract.info:eu-repo/semantics/publishedVersio

Phenolic composition and biological properties of cynara cardunculus l. Var. altilis petioles: Influence of the maturity stage

Hydroethanolic extracts of cardoon petioles collected at sixteen growth stages (P1–P16) were characterized in terms of their phenolic composition and bioactive potential (antioxidant, cytotoxic, anti-inflammatory, and antimicrobial activities). Fifteen phenolic compounds were tentatively identified (i.e., ten phenolic acids and five flavonoid glycosides); the main compounds were 5-O-caffeoylquinic and 1,5-di-O-caffeoylquinic acids. Samples collected at early maturity (P1–P4) presented a weak positive correlation between the higher content in polyphenols (P3: 101-mg/g extract) and better inhibition capacity against thiobarbituric acid reactive substance formation (TBARS; P3: IC50 = 5.0 µg/mL). Samples at intermediate maturation stages (P9) presented higher cytotoxic and anti-inflammatory potential. Moreover, immature petioles showed greater antihemolytic (OxHLIA; P4: IC50 = 65 and 180 µg/mL for Δt of 60 and 120 min, respectively) and antibacterial activity. The antifungal activity varied depending on the maturation stage and the fungi strain. In conclusion, the maturation stage may greatly affect the polyphenols composition and content and the bioactive potential of cardoon petioles.The authors are grateful to the Foundation for Science and Technology (FCT; Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020), for the F. Mandim PhD grant (SFRH/BD/146614/2019), and the J. Pinela (CEECIND/01001/2018) and M.I. Dias and L. Barros contracts through individual and institutional scientific employment program contracts. The authors are also grateful to the project TRANSCoLAB (0612_TRANS_CO_LAB_2_P), to the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project GreenHealth, Norte-01-0145-FEDER-000042, and to the Ministry of Education, Science and Technological Development of the Republic of Serbia (451-03-68/2020-14/200007). GIPUSAL was funded by Junta de Castilla y León (Spain) through Project SA093P20 and the Strategic Research Program for Units of Excellence (ref. CLU-2018-04).info:eu-repo/semantics/publishedVersio

Seasonal variation in bioactive properties and phenolic composition of cardoon (Cynara cardunculus var. altilis) bracts

Cardoon (Cynara cardunculus L.) bracts were collected at different maturation stages to investigate seasonal changes in the phenolic compounds profile and in vitro bioactivities. Among the 12 phenolic compounds tentatively identified, 3,5-O-dicaffeoylquinic acid (21.83 mg/g extract) and apigenin-7-O-glucuronide (10.6 mg/g extract) were the most abundant. Immature bracts (C1: principal growth stage (PGS) 5) had the highest phenolic compounds content, and anti-inflammatory (IC50 = 72 µg/mL) and cytotoxic (GI50 of 30–79 µg/mL) activities. Moreover, extract C1 inhibited efficiently the formation of thiobarbituric acid reactive substances (TBARS; IC50 = 26.8 µg/mL), while extract C8 (PGS 8/9) was more effective against oxidative haemolysis (IC50 38 and 75 µg/mL). The highest antibacterial and antifungal activities were attributed to samples C1 and C6 (PGS 7/8) and samples C2 (PGS 5/6) and C4 (PGS 6/7), respectively. Overall, the obtained results suggest the seasonal changes of polyphenolic composition and bioactivity of cardoon bracts of variable maturity.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020). National funding by FCT, through the institutional scientific employment program-contract with M.I. Dias, J. Pinela, and L. Barros. To FCT for the PhD grant (SFRH/BD/146614/2019) of F. Mandim. The authors are also grateful to FEDERInterreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E; to the project TRANSCoLAB (0612_TRANS_CO_LAB_2_P), and to the Ministry of Education, Science and Technological Development of the Republic of Serbia (451-03-68/2020-14/200007).info:eu-repo/semantics/publishedVersio

Valorization of Punica granatum L. leaves extracts as a source of bioactive molecules

Due to a lack of innovative valorization strategies, pomegranate processing generates a significant amount of residues with a negative environmental footprint. These by-products are a rich source of bioactive compounds with functional and medicinal benefits. This study reports the valorization of pomegranate leaves as a source of bioactive ingredients using maceration, ultrasound, and microwave-assisted extraction techniques. The phenolic composition of the leaf extracts was analyzed using an HPLC-DAD-ESI/MSn system. The extracts’ antioxidant, antimicrobial, cytotoxic, anti-inflammatory, and skin-beneficial properties were determined using validated in vitro methodologies. The results showed that gallic acid, (-)-epicatechin, and granatin B were the most abundant compounds in the three hydroethanolic extracts (between 0.95 and 1.45, 0.7 and 2.4, and 0.133 and 3.0 mg/g, respectively). The leaf extracts revealed broad-spectrum antimicrobial effects against clinical and food pathogens. They also presented antioxidant potential and cytotoxic effects against all tested cancer cell lines. In addition, tyrosinase activity was also verified. The tested concentrations (50–400 µg/mL) ensured a cellular viability higher than 70% in both keratinocyte and fibroblast skin cell lines. The obtained results indicate that the pomegranate leaves could be used as a low-cost source of value-added functional ingredients for potential nutraceutical and cosmeceutical applications.Ministerio de Ciencia e Innovación | Ref. RYC-2017-22891Ministerio de Ciencia e Innovación | Ref. FJC2019-042549-IMinisterio de Ciencia e Innovación | Ref. FJC20120-045405-IFundação para a Ciência e a Tecnologia | Ref. UIDB/00690/2020Fundação para a Ciência e a Tecnologia | Ref. UIDP/00690/2020Fundação para a Ciência e a Tecnologia | Ref. LA/P/0007/2021Fundação para a Ciência e a Tecnologia | Ref. SFRH/BD/146614/201