Chemical characterization and evaluation of biological activity of Cynara cardunculus extractable compounds

The Mediterranean species Cynara cardunculus L. is recognized in the traditional medicine, for their hepatoprotective and choleretic effects. Biomass of C. cardunculus L. var. altilis (DC), or cultivated cardoon, may be explored not only for the production of energy and pulp fibers, but also for the extraction of bioactive compounds. The chemical characterization of extractable components, namely terpenic and phenolic compounds, may valorize the cultivated cardoon plantation, due to their antioxidant, antitumoral and antimicrobial activities. In this study, the chemical composition of lipophilic and phenolic fractions of C. cardunculus L. var. altilis (DC), cultivated in the south of Portugal (Baixo Alentejo region) was characterized in detail, intending the integral valorization of its biomass. The biological activity of cultivated cardoon extracts was evaluated in terms of antioxidant, human tumor cell antiproliferative and antibacterial effects. Gas chromatography-mass spectrometry (GC-MS) was used for the chemical analysis of lipophilic compounds. Sixty-five lipophilic compounds were identified, from which 1 sesquiterpene lactone and 4 pentacyclic triterpenes were described, for the first time, as cultivated cardoon components, such as: deacylcynaropicrin, acetates of β- and α-amyrin, lupenyl acetate and ψ-taraxasteryl acetate. Sesquiterpene lactones were the major family of lipophilic components of leaves (≈94.5 g/kg), mostly represented by cynaropicrin (≈87.4 g/kg). Pentacyclic triterpenes were also detected, in considerably high contents, in the remaining parts of cultivated cardoon, especially in the florets (≈27.5 g/kg). Taraxasteryl acetate was the main pentacyclic triterpene (≈8.9 g/kg in florets). High pressure liquid chromatography-mass spectrometry (HPLC-MS) was utilized for the chemical analysis of phenolic compounds. Among the identified 28 phenolic compounds, eriodictyol hexoside was reported for the first time as C. cardunculus L. component, and 6 as cultivated cardoon components, namely 1,4-di-O-caffeoylquinic acid, naringenin 7-O-glucoside, naringenin rutinoside, naringenin, luteolin acetylhexoside and apigenin acetylhexoside. The highest content of the identified phenolic compounds was observed in the florets (≈12.6 g/kg). Stalks outer part contained the highest hydroxycinnamic acids abundance (≈10.3 g/kg), and florets presented the highest flavonoids content (≈10.3 g/kg). The antioxidant activity of phenolic fraction was examined through 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay. Stalks outer part, and receptacles and bracts extracts demonstrated the highest antioxidant effect on DPPH (IC50 of 34.35 μg/mL and 35.25 μg/mL, respectively). (cont.) abstract (cont.) The DPPH scavenging effect was linearly correlated with the total contents of hydroxycinnamic acids (r = -0.990). The in vitro antiproliferative activity of cultivated cardoon lipophilic and phenolic extracts was evaluated on a human tumor cells line of triple-negative breast cancer (MDA-MB-231), one of the most refractory human cancers to conventional therapeutics. After 48 h of exposition, leaves lipophilic extract showed higher inhibitory effect (IC50 = 10.39 μg/mL) than florets lipophilic extract (IC50 = 315.22 μg/mL), upon MDA-MB-231 cellular viability. Pure compound of cynaropicrin, representative of the main compound identified in leaves lipophilic extract, also prevented the cell proliferation of MDA-MB-231 (IC50 = 17.86 μM). MDA-MB-231 cells were much more resistant to the 48 h- treatment with phenolic extracts of stalks outer part (IC50 = 3341.20 μg/mL) and florets (IC50 > 4500 μg/mL), and also with the pure compound of 1,5-di-O-caffeoylquinic acid (IC50 = 1741.69 μM). MDA-MB-231 cells were exposed, for 48 h, to the respective IC50 concentrations of leaves lipophilic extract and pure compound of cynaropicrin, in order to understand their ability in modelling cellular responses, and consequently important potentially signaling pathways for the cellular viability decrease. Leaves lipophilic extract increased the caspase-3 enzymatic activity, contrarily to pure compound of cynaropicrin. Additionally, leaves lipophilic extract and pure compound of cynaropicrin caused G2 cell cycle arrest, possibly by upregulating the p21Waf1/Cip1 and the accumulation of phospho-Tyr15-CDK1 and cyclin B1. The inhibitory effects of leaves lipophilic extract and cynaropicrin pure compound, against the MDA-MB-231 cell proliferation, may also be related to the downregulation of phospho-Ser473-Akt. The antibacterial activity of cultivated cardoon lipophilic and phenolic extracts was assessed, for the first time, on two multidrug-resistant bacteria, such as the Gram-negative Pseudomonas aeruginosa PAO1 and the Gram-positive methicillin-resistant Staphylococcus aureus (MRSA), two of the main bacteria responsible for health care-associated infections. Accordingly, the minimum inhibitory concentrations (MIC) were determined. Lipophilic and phenolic extracts of florets did not have antibacterial activity on P. aeruginosa PAO1 and MRSA (MIC > 2048 μg/mL). Leaves lipophilic extract did not prevent the P. aeruginosa PAO1 growth, but pure compound of cynaropicrin was slightly active (MIC = 2048 μg/mL). Leaves lipophilic extract and pure compound of cynaropicrin blocked MRSA growth (MIC of 1024 and 256 μg/mL, respectively). The scientific knowledge revealed in this thesis, either by the chemical viewpoint, or by the biological viewpoint, contributes for the valorization of C. cardunculus L. var. altilis (DC) biomass. Cultivated cardoon has potential to be exploited as source of bioactive compounds, in conciliation with other valorization pathways, and Portuguese traditional cheeses manufacturing.

Uso de marcadores morfológicos, bioquímicos y moleculares SRAP para diferenciar variedades de Cynara cardunculus L. (Asteraceae)

Dentro del complejo primario de Cynara cardunculus L. se encuentran diversas variedades botánicas: var. scolymus (alcaucil), var. altilis (cardo cultivado) y var. sylvestris (cardo silvestre). A lo largo de la historia del mejoramiento, la caracterización de los materiales ha evolucionado desde el uso de caracteres morfológicos hasta los modernos análisis moleculares, pasando por los marcadores bioquímicos. El objetivo del presente trabajo fue comparar la utilidad de los marcadores morfológicos, bioquímicos y moleculares para la caracterización de materiales pertenecientes a tres variedades botánicas de Cynara cardunculus. Tres cultivares de la var. scolymus, dos de la var. altilis y dos de la var. sylvestris fueron caracterizadas por variables morfovegetativas, proteínas de reserva y por marcadores moleculares a través de la técnica SRAP. Estas metodologías permitieron discriminar dos grupos: uno incluyendo las variedades de cardo cultivado y silvestre, y el otro, las variedades del alcaucil. Los datos moleculares y morfológicos permitieron además diferenciar los cultivares evaluados de la var. scolymus. Se concluye que los marcadores analizados son útiles para la caracterización intravarietal e intervarietal en programas de mejoramiento.

Serpa cheese: technological, biochemical and microbiological characterisation of a PDO ewe's milk cheese coagulated with Cynara cardunculus L

Portugal has a strong tradition of cheesemaking from raw ewe's milk; most of these cheeses are still made on a traditional farmhouse scale. Their production is protected by Protected Designation of Origin (PDO) but the specific biochemical aspects of the majority still need to be characterised. Two different cheesemaking procedures, traditional and semi-industrial, were compared technologically, biochemically and microbiologically. It was observed that, despite the highly significant difference between artisanal and semi-industrial cheeses (P < 0.001), both products were within the limits of national regulations for most parameters except maturation temperature, humidity and the value for the maturation index. Although the present study was not fully representative of the region, the results obtained suggest that the specific regulations for Serpa cheese should be revised and that other parameters, such as moisture and salt-in-moisture content, which are very much dependent on the cheesemaking process, should be included in order to characterise better this traditional cheese.

Influência de genótipos de couve (Brassica oleracea L. var. acephala DC.) na biologia de Plutella xylostella (L., 1758) (Lepidoptera: Plutellidae)

Objetivou-se, com este trabalho, avaliar a influência de alguns genótipos de couve (Brassica oleracea L. var. acephala DC.) no desenvolvimento de Plutella xylostella (L., 1758) (Lepidoptera: Plutellidae). Os genótipos avaliados foram: Manteiga de Ribeirão Pires I-2620, Roxa I-919, Manteiga de São José, Manteiga de Monte Alegre, Pires 2 de Campinas, Couve Comum, Couve de Arthur Nogueira 2, Couve de Arthur Nogueira 1. Lagartas recém-eclodidas foram mantida em discos foliares de 8 cm de diâmetro para cada genótipo. Foram analisados os seguintes parâmetros: duração e viabilidade das fases larval e pupal, longevidade e fecundidade de adultos, utilizando análises paramétricas e de agrupamentos para interpretação dos dados. Observou-se um prolongamento em dias no ciclo de P. xylostella, aumento no peso de pupa e maiores valores de viabilidade e fecundidade, durante a segunda geração. O genótipo Couve de Arthur Nogueira 2 foi menos favorável ao desenvolvimento de P. xylostella nas duas gerações, e Couve Comum demonstrou maior influência negativa ao inseto na segunda geração. Manteiga de Ribeirão Pires I-2620 foi o mais suscetível nas duas gerações, agrupando com este na segunda geração Pires 2 de Campinas e Manteiga de São José.

Determinación de la capacidad parasitaria de la microbiota fúngica y de sus extractos acuosos en las semillas de cardo (Cynara cardunculus L.)

El presente estudio evalúa el efecto que 6 diferentes géneros hongos aislados a partir de semillas de 54 diferentes cultivares de cardo y sus extractos acuosos tienen sobre la germinación y nascencia de las semillas. Se han realizado pruebas de patogenicidad con dos aislados de cada uno de los seis géneros de mayor frecuencia del inventario (Aspergillus, Penicillium, Fusarium, Rhizopus, Cladosporium y Alternaria), así como de los extractos producidos tras 3, 7 y 11 días de incubación de los micetos. Los resultados de las inoculaciones con los micetos muestran efectos negativos sobre los porcentajes de germinación, con reducciones en la germinación que fueron máximas tras las inoculaciones con Rhizopus stolonifer (29% de disminución) y Fusarium verticillioides (23%). Los porcentajes de emergencia disminuyen tras duplicar la concentración del inóculo, aumentando además drásticamente el número de plántulas dañadas sobre el total de las emergidas. En el significativo caso de la inoculación con Cladosporium la duplicación del inóculo disminuyó la germinación hasta en un 31% respecto al testigo. Las plántulas emergidas tras las inoculaciones con los extractos obtenidos a partir de cultivos líquidos de los hongos ensayados presentaban los mismos síntomas de atrofias y daños sobre raíz y coleóptilo que los descritos para cada hongo. Los extractos acuosos de los géneros estudiados disminuyen también la germinación. Los resultados nos muestran la diferente capacidad parasitaria de cada una de las especies estudiadas apreciándose además diferencias según los diferentes periodos de agitación de los hongos y permiten asegurar que la producción de toxinas está regulada por el hongo, y que no aumenta linealmente con el crecimiento miceliar.

Inventario fúngico asociado a las semillas de cultivares de cardo (Cynara cardunculus L.)

Se caracterizó la micoflora presente en las semillas de 54 cultivares de cardo cosechados entre las campañas 1992-93 y 1997-98. 10 muestras procedentes del Servicio de Investigación Agraria, finca "La orden" (Badajoz) y 44 muestras de la Unidad Docente de Botánica de la Escuela Técnica Superior de Ingenieros Agrónomos de Madrid. Se analizaron un total de 4400 semillas, identificándose 15 géneros fúngicos diferentes. La presencia de los diferentes géneros varió en función de las localidades y los años de cosecha, entre la microbiota fúngica aislada cabe destacar por su potencial patogenicidad especies de los géneros Fusarium, Aspergillus y Alternaria.

Aprovechamiento del cardo (Cynara cardunculus L.) para la producción de biomasa lignocelulósica, aceite y forraje verde

Aprovechamiento del cardo (Cynara cardunculus L.) es una especie perteneciente a la familia de las Compuestas (Asteraceae) lo mismo que el cártamo o el girasol. es una especie vivaz gracias a sus yemas de recambio que posee en el cuello de la raíz estando muy bien adaptada a las condiciones del clima mediterráneo de veranos secos y calurosos.

Estudo da imobilização das proteases aspárticas da flor de Cynara cardunculus L. Reologia da coalhada

Mestrado em Biotecnologia, Faculdade de Engenharia de Recursos Naturais, Univ. do Algarve, 2005

Não-preferência para oviposição, alimentação e antibiose de Plutella xylostella (L., 1758) (Lepidoptera: Plutellidae) por genótipos de couve (Brassica oleracea L. var. acephala D.C.)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Atividade anti-inflamatória e citotoxicidade dos extratos glicólicos de Cynara cardunculus var. scolymus (L.) Fiori (alcachofra), Myracrodruon urundeuva Allemão (aroeira-do-sertão) e Camellia sinensis (L.) Kuntze (chá verde)

Pós-graduação em Biopatologia Bucal - ICT

Producción de primicia en invernadero de híbridos de espárrago verde (Asparagus officinalis var. Altilis) en su cuarto año productivo

Resumen: La producción de espárragos en Argentina está caracterizada por una elevada estacionalidad concentrada en el periodo octubre-diciembre, por lo que el empleo de invernaderos brinda la posibilidad de ampliar el calendario de oferta, anticipando la entrada en producción. Con el objetivo de evaluar el rendimiento de primicia de diferentes híbridos de espárrago verde, se realizó un ensayo en invernadero con ocho híbridos, en UCA Buenos Aires, iniciado el 15/11/2006, mediante plantines de 100 días a 1m*0,3m. Se evaluaron los siguientes genotipos: Italo, Zeno, Eros, Ercole, H-668, Marte, y Giove, de origen italiano, y UC-157 de origen americano, como testigo por ser el tradicionalmente cultivado en Argentina. Se evaluaron 22 cosechas, con una frecuencia de día por medio, del 17/08/2011-25/10/2011. Se estudiaron diferencias en kg totales y comerciales (PFT y PFC), Nº turiones totales y comerciales/ha (NTT y NTC) y distribución de calibres (DC): Jumbo (J), Extra-Large (XL), Large (L), Medium (M), Small (S) y Asparagina (A). Se efectuó un análisis multifactor ANOVA LSD test (P>0.05). En promedio se obtuvieron: PFT: 17053; PFC: 7904 kg.ha-1; NTT: 670566 y NTC: 520938 turiones.ha-1. Se destacaron: en PFT: Italo: 29458a, Zeno: 23056b, Giove: 23034b y H-668: 18568bc; en PFC: Italo: 14850a, Giove: 9856b, Zeno: 9130bc y H-668: 8228bcd; en NTT: Italo: 842512a, H-668: 754512a, Giove: 715000b y Eros: 707498b; en NTC: Italo: 844998a, H-668: 667502b y Eros: 542498bc y en DC: en J: Italoa, Gioveb, Zenobc y UC-157bcd; en XL: Italoa, Zenoab y Gioveb; en L: Giovea, Italoa, Zenoab, H-668abc y UC-157abc; en M: Italoa, H-668ab y Erosab; en S: H-668a, Erosa e Italoab y en A: H-668a y Marteab. Por lo expuesto resulta alentadora la productividad de Italo, Giove, Zeno y H-668 para producción de espárrago verde de primicia en invernadero.

Efeito do tamanho de tubetes na qualidade de mudas de Jatobá (Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang.), Ipê-amarelo (Tabebuia chrysotricha (Mart. ex DC.) Sandl.) e Guarucaia (Parapiptadenia rigida (Benth.) Brenan)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

A study of the morphological variability in Cynara humilis L. and C. Hystrix BalI (Aseraceae-Cardueae)

El estudio de los especimenes de herbario, recolectados en la Península Ibérica y Marruecos, y que han sido atribuidos a Cvttara humilis L y C hystrtc Raíl ha puesto de manifiesto la variabilidad de ciertos caracteres, mucho más notable en eí material marroquí. La representación mediante símbolos, en un mapa, de la distribución de seis dc los caracteres morfológicos seleccionados, permite comprobar la existencia de formas intermedias, que se situan en la zona de contacto entre las áreas marroquíes dc ambos túxones. Sc discute la posibilidad de explicar este hecho, como resultado de un proceso de hibridación alopátrica introgresiva.

Urana subterranean clover (Trifolium subterraneum L. var. subterraneum)

Urana is a hardseeded, moderately early flowering F-5-derived crossbred subterranean clover of var. subterraneum [( Katz. et Morley) Zohary and Heller] developed by the collaborating organisations of the National Annual Pasture Legume Improvement Program. It has been selected for release as a new cultivar on the basis of its high winter and spring herbage production and overall field performance relative to other subterranean clovers of similar maturity. Urana is recommended for sowing in Western Australia, New South Wales, Victoria, South Australia and Queensland. It is best suited to well-drained, moderately acidic soils in areas with a growing season of 5 - 7 months, which extends into mid-October. Urana is suited to phase farming and crop rotations. It has been granted Plant Breeders Rights in Australia.

Coolamon subterranean clover (Trifolium subterraneum L. var. subterraneum)

Coolamon is a mid-season to late-season flowering F4-derived crossbred subterranean clover of var. subterraneum, developed by the collaborating organisations of the National Annual Pasture Legume Improvement Program. It is a replacement for Junee and has been selected for release on the basis of its greater herbage production and persistence, and its resistance to both known races of clover scorch. Coolamon is recommended for sowing in Western Australia, New South Wales, Victoria, South Australia and Queensland. It is best suited to well-drained, moderately acidic soils in areas with a growing season of 6.5-8 months that extends into November. Coolamon is best suited to phase farming and permanent pasture systems. It can also be used in cropping rotations, but at least 2 years of pasture are required between crops. Coolamon has been granted Plant Breeders Rights in Australia.