39 research outputs found
Water Extract of Cryphaea heteromalla (Hedw.) D. Mohr Bryophyte as a Natural Powerful Source of Biologically Active Compounds
Bryophytes comprise of the mosses, liverworts, and hornworts. Cryphaea heteromalla,
(Hedw.) D. Mohr, is a non-vascular lower plant belonging to mosses group. To the date, the most
chemically characterized species belong to the liverworts, while only 3.2% and 8.8% of the species
belonging to the mosses and hornworts, respectively, have been investigated. In this work, we present
Folin–Ciocalteu and oxygen radical absorbance capacity (ORAC) data related to crude extracts of C.
heteromalla obtained by three di erent extraction solvents: pure water (WT), methanol:water (80:20
v/v) (MET), and ethanol:water (80:20 v/v) (ETH). The water extract proved to be the best solvent
showing the highest content of biophenols and the highest ORAC value. The C. heteromalla-WT
extract was investigated by HPLC-TOF/MS (High Performance Liquid Chromatography-Time of
Flight/Mass Spectrometry) allowing for the detection of 14 compounds, five of which were phenolic
compounds, derivatives of benzoic, caffeic, and coumaric acids. Moreover, the C. heteromalla WT
extract showed a protective effect against reactive oxygen species (ROS) generation induced by
tert-butyl hydroperoxide (TBH) on the murine NIH-3T3 fibroblast cell line.This work was supported by the European Social Funding (FSE Sicilia 2020 Project ID: 2014.IT.05.SFOP.014/
3/10.4/9.2.10/0014)
Revalorization of Broccoli By-Products for Cosmetic Uses Using Supercritical Fluid Extraction
The agri-food industry is currently one of themain engines of economic developmentworldwide.
The region ofMurcia is a reference area in Europe for the cultivation of fruits and vegetables and produces
the bulk of Spanish exports of broccoli (Brassica oleracea var. italica). The processing of fresh produce
generates a huge number of by-products that represent an important economic and environmental problem
when discarded. In this work, an advanced extraction technique using environmentally friendly solvents
was applied to assess the revalorization of broccoli by-products, by performing a comparative analysis
with conventional extraction. To achieve this goal, supercritical fluid extraction based on response surface
methodology was performed using CO2 and ethanol as solvents. The results obtained showed that the
supercritical fluid extracts were rich in -carotene, phenolic compounds, chlorophylls and phytosterols.
Moreover, in bioactivity assays, the supercritical fluid extracts exhibited a high antioxidant activity and a
cytoprotective effect in a non-tumorigenic keratinocyte cell line exposed to ultraviolet B light. The results
indicate that supercritical fluid extracts from broccoli by-products could potentially serve as an ingredient
for cosmetic purposes
HPLC-ESI-TOF-MS chemical characterization of comfrey root extract obtained by subcritical water extraction
In this work, a study on phytochemical profiles of comfrey (Symphytum officinale L.) root extract obtained by subcritical water extraction (SWE) has been carried out. Chemical composition was assessed by high-performance liquid chromatography coupled with electrospray time-of-flight mass spectrometry (HPLC-ESI-TOF-MS) identifying 22 compounds including organic acids, phenolic acids, flavonoids and fatty acids. Great number of phenolic acids and flavonoids were found in the extract obtained by SWE, with citric acid, caffeic acid and derivative, salvianolic acid B, hydroxybenzoic acid, syringetin-3-O-glucoside and quercetin 3-O-malonylglucoside as the most abundant compounds. Moreover, quercitin-3- malonylglucoside isomers, hydroxybenzoic acid glucoside, cirismaritin isomers, p-coumaric acid, hydroxycoumarin and methylcoumarin, among others, were identified for the first time in S. officinale root. Overall, the results indicate the potential of SWE for the production of highquality plant extracts from S. officinale root
Identification of bioactive compounds in comfrey root extracts
In the present study bioactive compounds present in comfrey root extracts obtained by supercritical fluid (SFE) and pressurized liquid extraction (PLE) were identified. Chemical characterization of the extracts was carried out by high-performance liquid chromatography coupled to DAD and electrospray-ionization time-of-flight mass spectrometry (HPLC–ESITOF-MS) yielding in total of 23 identified compounds. PLE as a fast, green and innovative approach, seems to be the best choice for extracting wide variety of compounds with different polarities within the shortest extraction time being the fatty acids and their derivatives the most abundant. The present study also highlights the potential application of comfrey root extracts as constituents of new added-value formulations
Permeability Study of Polyphenols Derived from a Phenolic-Enriched Hibiscus sabdariffa Extract by UHPLC-ESI-UHR-Qq-TOF-MS
Previous findings on the capacity of Hibiscus sabdariffa (HS) polyphenols to ameliorate metabolic disturbances justify the necessity of studies oriented to find the potential metabolites responsible for such an effect. The present study examined the intestinal epithelial membrane permeability of polyphenols present in a phenolic-enriched Hibiscus sabdariffa extract (PEHS), free and encapsulated, using the Caco-2 cell line. Additionally, selected polyphenols (quercetin, quercetin-3-glucoside, quercetin-3-glucuronide, and N-feruloyltyramine) were also studied in the same absorption model. The powerful analytical platform used ultra-high-performance liquid chromatography coupled with ultra-high-resolution quadrupole time-of-flight mass spectrometry (UHPLC-ESI-UHR-Qq-TOF-MS), and enabled the characterization of seven new compounds in PEHS. In the permeation study, only a few compounds were able to cross the cell monolayer and the permeability was lower when the extract was in an encapsulated form. Pure compounds showed a moderate absorption in all cases. Nevertheless, these preliminary results may need further research to understand the complete absorption mechanism of Hibiscus polyphenols.This work was supported by projects AGL2011-29857-C03-02 and AGL2011-29857-C03-03 (Spanish Ministry of Science and Innovation), in addition to P10-FQM-6563 and P11-CTS-7625 (Andalusian Regional Government Council of Innovation and Science), PROMETEO/2012/007 and ACOMP/2013/093 (Generalitat Valenciana), and CIBER (CB12/03/30038, Fisiopatología de la Obesidad y la Nutrición, CIBERobn, Instituto de Salud Carlos III). The authors are grateful to the Spanish Ministry of Science and Innovation for the grant FPI (BES-2009-028128), and the Spanish Ministry of Economy and Competitiveness (MINECO) in association with the European Social Fund (FSE) for the contract PTQ-13-06429. MH is a recipient of a VALi + D fellowship from GV (ACIF/2010/162). The authors are especially grateful to Bruker Daltonik GmbH (Bremen, Germany) for their help and support during this research
Cocoa and Grape Seed Byproducts as a Source of Antioxidant and Anti-Inflammatory Proanthocyanidins
Phenolic compounds, which are secondary plant metabolites, are considered an integral part of the human diet. Physiological properties of dietary polyphenols have come to the attention in recent years. Especially, proanthocyanidins (ranging from dimers to decamers) have demonstrated potential interactions with biological systems, such as antiviral, antibacterial, molluscicidal, enzyme-inhibiting, antioxidant, and radical-scavenging properties. Agroindustry produces a considerable amount of phenolic-rich sources, and the ability of polyphenolic structures to interacts with other molecules in living organisms confers their beneficial properties. Cocoa wastes and grape seeds and skin byproducts are a source of several phenolic compounds, particularly mono-, oligo-, and polymeric proanthocyanidins. The aim of this work is to compare the phenolic composition of Theobroma cacao and Vitis vinifera grape seed extracts by high pressure liquid chromatography coupled to a quadrupole time-of-flight mass spectrometer and equipped with an electrospray ionization interface (HPLC-ESI-QTOF-MS) and its phenolic quantitation in order to evaluate the proanthocyanidin profile. The antioxidant capacity was measured by different methods, including electron transfer and hydrogen atom transfer-based mechanisms, and total phenolic and flavan-3-ol contents were carried out by Folin–Ciocalteu and Vanillin assays. In addition, to assess the anti-inflammatory capacity, the expression of MCP-1 in human umbilical vein endothelial cells was measured.This work was supported by the project AGL2011-29857-C03-02 and BFU2014-52433-C3-2-R (Spanish Ministry of Science and Innovation), as well as P10-FQM-6563 and P11-CTS-7625 (Andalusian Regional Government Council of Innovation and Science). The author Isabel Borrás Linares acknowledges financial support from the Spanish Ministry of Economy and Competitiveness (MINECO) and the European Social Fund (FSE) for the contract PTQ-13-06429. Authors are also grateful to the Universitat Rovira I Virgili for the Martí I Franquès Grant 2016PMF-POST-02 awarded to Salvador Fernández-Arroyo
Aprendizaje invertido- Aprendizaje colaborativo, Flipped learning-Jigsaw, en el laboratorio y en el aula universitaria.
El aula invertida o “flipped classroom”, y la técnica del puzle o “Jigsaw” son metodologías pedagógicas cuyo principal objetivo consiste en que el alumno/a asuma un rol más activo en su proceso de aprendizaje que el que venía ocupando tradicionalmente. Los objetivos que se pretenden alcanzar son: mejorar los resultados de aprendizaje, fomentar el pensamiento crítico, el trabajo autónomo del estudiante, así como el trabajo colaborativo y la participación activa del estudiante en el proceso de aprendizaje. La metodología didáctica a seguir se explicará previamente a todos los estudiantes implicados. Se dejan algunas sesiones de laboratorio y seminarios teórico-prácticos para desarrollarlos siguiendo la metodología tradicional, considerándolos sesiones de control para poder comparar los resultados finales obtenidos con la metodología propuesta. Hasta la fecha la metodología seguida en las sesiones prácticas de laboratorio se limitaba a leer un guion de prácticas y seguir unas instrucciones, en las sesiones de seminarios numéricos o casos teórico-prácticos, de forma similar, el profesor dedica varias sesiones a poner en contexto a los estudiantes. El problema de esta metodología, es que muchas veces los estudiantes tienen dificultades para asimilar, en una sola sesión de clase, los conceptos teóricos que tienen que poner en práctica, ya que están más preocupados por seguir las instrucciones, que por comprender el porqué de lo que están haciendo. Para solucionar estos inconvenientes se propone esta metodología docente.
The "flipped classroom" and the "Jigsaw" techniques are pedagogical methodologies whose main objective is for students to take on a more active role in the learning process than they have traditionally done. The objectives to be achieved are: to improve learning results, encourage critical thinking, autonomous student work, collaborative work, and the active participation of students in the learning process. The didactic methodology will be explained previously to the students. Some laboratory sessions and theoretical and practical seminars are left to be developed following the traditional methodology, considering them as control sessions to compare the final results obtained between both didactic methodologies. Until now, the methodology followed in the practical laboratory sessions was limited to reading a practical script and following instructions, while in the numerical seminar sessions or theoretical-practical cases, the teacher dedicates several sessions to putting the students in context. The problem with this methodology is that students often find it difficult to assimilate, in a single class session, the theoretical concepts they have to put into practice, as they are more concerned with following instructions than with understanding why they are doing what they are doing. This teaching methodology is proposed to solve these problems.Proyecto de Innovación docente y Buenas Prácticas Docentes. Plan Fido. Proyecto 22-9
Rosmarinus Officinalis Leaves as a Natural Source of Bioactive Compounds
In an extensive search for bioactive compounds from plant sources, the composition of different extracts of rosemary leaves collected from different geographical zones of Serbia was studied. The qualitative and quantitative characterization of 20 rosemary (Rosmarinus officinalis) samples, obtained by microwave-assisted extraction (MAE), was determined by high performance liquid chromatography coupled to electrospray quadrupole-time of flight mass spectrometry (HPLC–ESI-QTOF-MS). The high mass accuracy and true isotopic pattern in both MS and MS/MS spectra provided by the QTOF-MS analyzer enabled the characterization of a wide range of phenolic compounds in the extracts, including flavonoids, phenolic diterpenes and abietan-type triterpenoids, among others. According to the data compiled, rosemary samples from Sokobanja presented the highest levels in flavonoids and other compounds such as carnosol, rosmaridiphenol, rosmadial, rosmarinic acid, and carnosic acid. On the other hand, higher contents in triterpenes were found in the extracts of rosemary from Gložan (Vojvodina).This work was financially supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia through a national project (Project Number TR 31014) and through scientific and technological cooperation between the Republic of Serbia and Spain (Project Number RE451-03-02635/2011-14/4). The authors are also grateful to the Spanish Ministry of Science and Innovation for the project AGL2011-29857-C03-02 and the grant FPI BES-2009-028128, Andalusian Regional Government Council of Innovation and Science for the Excellence Projects P09-CTS-4564, P10-FQM-6563 and P11-CTS-762
Extra-virgin olive oil contains a metabolo-epigenetic inhibitor of cancer stem cells
We are grateful to
Custodio Borrego for giving us free use of the photograph he
took of EVOO and olive trees in Granada (Spain), which have
been included in Figure 7. This work has been awarded with the
IV Premio Internacional Castillo de Canena de Investigación Oleícola
‘LUIS VAÑÓ’(IV Edition of Castillo de Canena LUIS VAÑÓ Award for
Research on Olive Cultivation and Olive Oil; UC Davis Olive Center,
Castillo de Canena, and Universidad de Jaén).The authors would like to thank Dr Kenneth McCreath for editorial
support. We are greatly indebted to Prof Robert A. Weinberg
(Whitehead Institute for Biomedical Research, Cambridge, MA)
for providing the HMLERshCntrol/HMLERshEcad cells used in this
work.Targeting tumor-initiating, drug-resistant populations of cancer stem cells (CSC) with phytochemicals is a novel
paradigm for cancer prevention and treatment. We herein employed a phenotypic drug discovery approach coupled to
mechanism-of-action profiling and target deconvolution to identify phenolic components of extra virgin olive oil (EVOO)
capable of suppressing the functional traits of CSC in breast cancer (BC). In vitro screening revealed that the secoiridoid
decarboxymethyl oleuropein aglycone (DOA) could selectively target subpopulations of epithelial-like, aldehyde
dehydrogenase (ALDH)-positive and mesenchymal-like, CD44+CD24−/low CSC. DOA could potently block the formation of
multicellular tumorspheres generated from single-founder stem-like cells in a panel of genetically diverse BC models.
Pretreatment of BC populations with noncytotoxic doses of DOA dramatically reduced subsequent tumor-forming capacity
in vivo. Mice orthotopically injected with CSC-enriched BC-cell populations pretreated with DOA remained tumor-free for
several months. Phenotype microarray-based screening pointed to a synergistic interaction of DOA with the mTOR inhibitor
rapamycin and the DNA methyltransferase (DNMT) inhibitor 5-azacytidine. In silico computational studies indicated that
DOA binds and inhibits the ATP-binding kinase domain site of mTOR and the S-adenosyl-l-methionine (SAM) cofactorbinding
pocket of DNMTs. FRET-based Z-LYTE™ and AlphaScreen-based in vitro assays confirmed the ability of DOA to
function as an ATP-competitive mTOR inhibitor and to block the SAM-dependent methylation activity of DNMTs. Our
systematic in vitro, in vivo and in silico approaches establish the phenol-conjugated oleoside DOA as a dual mTOR/DNMT
inhibitor naturally occurring in EVOO that functionally suppresses CSC-like states responsible for maintaining tumorinitiating
cell properties within BC populations.This work was supported by grants from the Ministerio de
Ciencia e Innovación (Grant SAF2016-80639-P to J.A.M.),
Plan Nacional de I+D+I, Spain, the Agència de Gestió d’Ajuts
Universitaris i de Recerca (AGAUR; Grant 2014 SGR229 to J.A.M.),
Departament d’Economia i Coneixement, Catalonia, Spain,
the Andalusian Regional Government Council of Innovation
and Science (Grant P11-CTS-7625 to A.S.-C.), the Ministerio de
Economía, Industria y Competitividad, Spain (Grants AGL2015-
67995-C2-3-R and AGL2015-67995-C3-1-R to A.S.-C. and V.M.)
and Conselleria d’Educació, Investigació, Cultura I Esport,
Generalitat Valenciana, Spain (Grant PROMETEO/2016/006 to
V.M). E.C. is supported by the Sara Borrell post doctoral contract
(CD15/00033) from the Ministerio de Sanidad y Consumo,
Fondo de Investigación Sanitaria (FIS), Spain
Comparison of different extraction procedures for the comprehensive characterization of bioactive phenolic compounds in Rosmarinus officinalis by reversed-phase high-performance liquid chromatography with diode array detection coupled to electrospray time-of-flight mass spectrometry
El pdf del artículo es la versión pre-print.-- et al.In the present work, a comparative study between two environmentally friendly and selective extraction techniques, such as supercritical fluid extraction (SFE) and pressurized liquid extraction (PLE) have been carried out focusing in the bioactive phenolic compounds present in Rosmarinus officinalis. For the analysis of the SFE and PLE extracts, a new methodology for qualitative characterization has been developed, based on the use of reversed-phase high-performance liquid chromatography (RP-HPLC), equipped with two different detection systems coupled in series: diode array detector (DAD) and time of flight mass spectrometry (TOF-MS) detector connected via an electrospray ionization interface (ESI). The use of a small particle size C18 column (1.8μm) provided a great resolution and made possible the separation of several isomers. Moreover, UV-visible spectrophotometry is a valuable tool for identifying the class of phenolic compounds, whereas MS data enabled to structurally characterize the compounds present in the extracts. The applied methodology was useful for the determination of many well-known phenolic compounds present in R. officinalis, such as carnosol, carnosic acid, rosmadial, rosmanol, genkwanin, homoplantaginin, scutellarein, cirsimaritin and rosmarinic acid, as well as other phenolic compounds present in other species belonging to Lamiaceae family. © 2011 Elsevier B.V.The author IBL acknowledges financial support from the Spanish Ministry of Education and Science (FPI grant, BES-2009-028128). M.H. would like to thank MICINN for a “Ramón y Cajal” contract. The authors are grateful to the Spanish Ministry of Education and Science for the project AGL2008-05108-C03-03/ALI and to Andalusian Regional Government Council of Innovation and Science for the excellence projects P07-AGR-02619, P09-CTS-4564 and P10-FQM-6563 and to CSD2007-00063 FUN-C-FOOD (Programa CONSOLIDER-INGENIO 2010) projects.Peer Reviewe