Silibinin Overcomes EMT-Driven Lung Cancer Resistance to New-Generation ALK Inhibitors

Epithelial-to-mesenchymal transition (EMT) may drive the escape of ALK-rearranged non-small-cell lung cancer (NSCLC) tumors from ALK-tyrosine kinase inhibitors (TKIs). We investigated whether first-generation ALK–TKI therapy-induced EMT promotes cross-resistance to new-generation ALK–TKIs and whether this could be circumvented by the flavonolignan silibinin, an EMT inhibitor. ALK-rearranged NSCLC cells acquiring a bona fide EMT phenotype upon chronic exposure to the first-generation ALK–TKI crizotinib exhibited increased resistance to secondgeneration brigatinib and were fully refractory to third-generation lorlatinib. Such cross-resistance to new-generation ALK–TKIs, which was partially recapitulated upon chronic TGF stimulation, was less pronounced in ALK-rearranged NSCLC cells solely acquiring a partial/hybrid E/M transition state. Silibinin overcame EMT-induced resistance to brigatinib and lorlatinib and restored their efficacy involving the transforming growth factor-beta (TGF )/SMAD signaling pathway. Silibinin deactivated TGF -regulated SMAD2/3 phosphorylation and suppressed the transcriptional activation of genes under the control of SMAD binding elements. Computational modeling studies and kinase binding assays predicted a targeted inhibitory binding of silibinin to the ATP-binding pocket of TGF type-1 receptor 1 (TGFBR1) and TGFBR2 but solely at the two-digit micromolar range. A secretome profiling confirmed the ability of silibinin to normalize the augmented release of TGF into the extracellular fluid of ALK–TKIs-resistant NSCLC cells and reduce constitutive and inducible SMAD2/3 phosphorylation occurring in the presence of ALK–TKIs. In summary, the ab initio plasticity along the EMT spectrum may explain the propensity of ALK-rearranged NSCLC cells to acquire resistance to new-generation ALK–TKIs, a phenomenon that could be abrogated by the silibinin-driven attenuation of the TGF /SMAD signaling axis in mesenchymal ALK-rearranged NSCLC cells.Ministry of Science and Innovation, Spain (MICINN) Spanish GovernmentPlan Nacional de l+D+I PID2019-10455GB-I00 CP20/00003 Spanish GovernmentFundacio Oncolliga Girona (Lliga catalana d'ajuda al malalt de cancer, Girona)Spanish GovernmentCenter for Forestry Research & Experimentation (CIEF)European Commission PI22/00297Grupo Espanol de Cancer de Pulmon (GECP) RTI2019-096724-B-C21La Marato de TV3 foundationHealth Research and Innovation Strategic Plan PROMETEO/2021/059Pla strategic de recerca i innovacio en salut 201906Generalitat de CatalunyaInstituto de Salud Carlos III SLT006/17/11

Nigella Plants – Traditional Uses, Bioactive Phytoconstituents, Preclinical and Clinical Studies

M. d. M. Contreras thanks the FEDER UJA project 1260905 funded by "Programa Operativo FEDER 2014-2020" and "Consejeria de Economia y Conocimiento de la Junta de Andalucia". This work was also supported by CONICYT PIA/APOYO CCTE AFB170007.Nigella is a small genus of the family Ranunculaceae, which includes some popular species due to their culinary and medicinal properties, especially in Eastern Europe, Middle East, Western, and Central Asia. Therefore, this review covers the traditional uses and phytochemical composition of Nigella and, in particular, Nigella sativa. The pharmacological studies reported in vitro, in vivo, and in humans have also been reviewed. One of the main strength of the use of Nigella is that the seeds are rich in the omega-6 fatty acid linoleic acid and provide an extra-source of dietary phytochemicals, including the bioactive thymoquinone, and characteristics saponins, alkaloids, and flavonoids. Among Nigella species, N. sativa L. is the most studied plant from the genus. Due to the phytochemical composition and pharmacological properties, the seed and seed oil from this plant can be considered as good candidates to formulate functional ingredients on the basis of folklore and scientific knowledge. Nonetheless, the main limations are that more studies, especially, clinical trials are required to standardize the results, e.g. to establish active molecules, dosage, chemical profile, long-term effects and impact of cooking/incorporation into foods.Programa Operativo FEDER 2014-2020 1260905Junta de Andalucia 1260905CONICYT PIA/APOYO CCTE AFB17000

Current Disease-Targets for Oleocanthal as Promising Natural Therapeutic Agent

The broad number of health benefits which can be obtained from the long-term consumption of olive oil are attributed mainly to its phenolic fraction. Many olive oil phenolics have been studied deeply since their discovery due to their bioactivity properties, such as Hydroxytyrosol. Similarly, in the last decade, the special attention of researchers has been addressed to Oleocanthal (OC). This olive oil phenolic compound has recently emerged as a potential therapeutic agent against a variety of diseases, including cancer, inflammation, and neurodegenerative and cardiovascular diseases. Recently, different underlying mechanisms of OC against these diseases have been explored. This review summarizes the current literature on OC to date, and focuses on its promising bioactivities against different disease-targets.This work was supported by the project of Spanish Ministry of Science and Innovation (AGL2015-67995-C3-2-R)

Sweet Cherry Byproducts Processed by Green Extraction Techniques as a Source of Bioactive Compounds with Antiaging Properties

Authors thank GRAFOR BIOTECH S.L. by its support to the project.In the cosmetic industry, there is a continuous demand for new and innovative ingredients for product development. In the context of continual renovation, both cosmetic companies and customers are particularly interested in compounds derived from natural sources due to their multiple benefits. In this study, novel and green-extractive techniques (pressurized solvent, supercritical CO2, and subcritical water extractions) were used to obtain three new extracts from sweet cherry stems, a byproduct generated by the food industry. The extracts were characterized by high-performance liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry (HPLC-ESI-QTOF-MS), and 57 compounds, mainly flavonoids but also organic and phenolic acids, fatty acids, and terpenes, were identified. After analytical characterization, a multistep screening approach, including antioxidant, enzymatic, and photoprotective cellular studies, was used to select the best extract according to its benefits of interest to the cosmetics industry. The extract obtained with supercritical CO2 presented the best characteristics, including a wide antioxidant capacity, especially against lipid peroxyl and •OH free radicals, as well as relevant photoprotective action and antiaging properties, making it a potential new ingredient for consideration in the development of new cosmetics.This research was funded by projects RTI2018-096724-B-C21 and 2018-096724-B-C22 from the Spanish Ministry of Science, Innovation and Universities; APOSTD/2018/097 fellowship from Generalitat Valenciana to M.H-.L. and CIBER (CB12/03/30038, Fisiopatología de la Obesidad y la Nutrición, CIBERobn)

Assessment of conventional and microwave heating effects on the variation of the bioactive compounds of Chétoui VOO using HPLC-DAD-ESI-TOF-MS

The goal of this work was to monitor the hydrophilic profile of the virgin olive oil (VOO) of the second main Tunisian variety, ‘‘Chétoui”, in order to achieve a better understanding of the behavior of the bioactive phenolic compounds during 0, 2, 5, 10, and 15 min of microwave heating at medium power 800 W, and 0, 2.5, and 5 h of conventional heating at 180 ºC. The extent of the oxidative and hydrolytic degradation of the different phenolic subclasses was evaluated using highperformance liquid chromatography (HPLC) coupled to electrospray time-of-flight mass spectrometry (TOF-MS) method. During heating process, the most represented component in Chétoui VOO was found to be isomer 1 and 2 of deacetoxy oleuropein aglycone, and hydroxy decarboxy oleuropein aglycon. These compounds may be considered as direct markers for the degree of transformation of secoiridoids during heating process. Among the studied phenolic compounds, hydroxytyrosol, tyrosol, luteolin and apigenin displayed the highest heating resistance in the whole time range of microwave and conventional applications. However, the main secoiridoids quantified in the fresh VOO, isomer 2 of oleuropein aglycone, and ligstroside aglycone, decreased in concentration with the thermal treatment and this decrease was drastic under conventional heating.This work was supported by the Ministry of High Education, and Scientific Research in Tunisia, and the Spanish Ministry of Economy and Competitiveness (MINECO) (AGL2015-67995- C3-2-R) and the Andalusian Regional Government Council (P11-CTS-7625)

Potential Hepatoprotective Activity of Super Critical Carbon Dioxide Olive Leaf Extracts against CCl4-Induced Liver Damage

Virgin olive oil has demonstrated its effective activity against oxidative stress. However, data on the bioactive effect of olive leaves or their major constituents on the liver are scarce. The present research work was conducted to evaluate the hepatoprotective effects of supercritical carbon dioxide (SC-CO2) extracts from fresh and dried olive leaves on hepatotoxicity caused by carbon tetrachloride (CCl4) in rat models. For this purpose, healthy albino rats of 180–250 g weight were used. The assessment of biochemical markers was carried out on blood and liver tissue. Then, a histopathological study was carried out on liver tissue. The obtained results showed that fresh and dried olive leaf extracts ameliorate the perturbed biochemical parameters caused by CCl4 treatment. Furthermore, the results registered for the histopathological study are in accordance with the biochemical parameters and the protective capacity of SC-CO2 extracts against DNA damage, indicating that olive leaf extracts helped to improve liver fibrosis caused by CCl4 treatment.Tunisian Ministry of Higher Education and Scientific Research LR15CBBC05Spanish Ministry of Economy and Competitiveness (MINECO) AGL2015-67995-C3-2Junta de Andalucia B-AGR-466-UGR1

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)

Extraction and Analysis of Phenolic Compounds in Rice: A Review

Rice represents the main source of calorie intake in many world countries and about 60% of the world population include rice in their staple diet. Whole grain rice, also called brown rice, represent the unpolished version of the more common white rice including bran, germ, and endosperm. Many health-promoting properties have been associated to the consumption of whole grain rice and, for this reason, great attention has been paid by the scientific community towards the identification and the quantification of bioactive compounds in this food item. In this contribution, the last five years progresses in the quali-quantitative determination of phenolic compounds in rice have been highlighted. Special attention has been devoted to the most recent strategies for the extraction of the target compounds from rice along with the analytical approaches adopted for the separation, identification and quantification of phenolic acids, flavonoids, anthocyanins, and proanthocyanidins. More specifically, the main features of the “traditional” extraction methods (i.e., maceration, ultrasound-assisted extraction) have been described, as well as the more innovative protocols involving advanced extraction techniques, such as MAE (microwave-assisted extraction). The predominant role of HPLC in the definition of the phenolic profile has been examined also presenting the most recent results obtained by using mass spectrometry-based detection systems. In addition, the most common procedures aimed to the quantification of the total amount of the cited classes of phenolic compounds have been described together with the spectrophotometric protocols aimed to the evaluation of the antioxidant properties of rice phenolic extracts (i.e., FRAP, DPPH, ABTS and ORAC)

Optimizing vacuum drying process of polyphenols, flavanols and DPPH radical scavenging assay in pod husk and bean shell cocoa

The objective of this study was to optimize different vacuum drying conditions for cocoa pod husk and cocoa bean shell in order to enhance these by-products for commercial applications. To carry out the optimization, the response surface methodology was applied using a Box–Behnken experimental design with 15 experiments for which different conditions of temperature (X1), drying time (X2) and vacuum pressure (X3) were established. The response variables were the content of total polyphenols, the content of flavanols and the radical scavenging activity evaluated in the extracts of the different experiments. Temperature (50–70 °C), drying time (3–12 h) and vacuum pressure (50–150 mbar) were considered as independent variables. The main factors affecting the response variables were temperature, followed by vacuum pressure. For the content of polyphenols, the optimal response values predicted for the cocoa pod husk was 11.17 mg GAE/g with a confidence limit (95%) of 9.05 to 13.28 mg GAE/g (optimal conditions: 65 °C, 8 h and 75 mbar), while for the cocoa bean shell cocoa was 29.61 mg GAE/g with a confidence limit (95%) of 26.95 to 32.26 mg GAE/g (optimal conditions: 50 °C, 5 h and 100 mbar). Therefore, results of this study suggest a high content of phenolic compounds obtained from these by-products that show relevance as functional ingredients for application in the food, nutraceutical, and cosmeceutical industriesFondo Nacional de Desarrollo Científico, Tecnológico y de Innovación Tecnológica of Perú, project number 184-2020-FONDECY

Diet Supplementation with Polyphenol-Rich Salicornia ramosissima Extracts Protects against Tissue Damage in Experimental Models of Cerebral Ischemia

Strokes are the secondmost common cause of death worldwide and a leading cause of disability. Regular consumption of polyphenols has been shown to reduce the risk of suffering a cardiovascular event. For this reason, we have investigated the protective effect of Salicornia ramosissima, a seasonal halophyte that synthetizes high amounts of bioactive compounds, including polyphenols, in response to environmental stress. Aqueous, hydroalcoholic, and ethanolic extracts were prepared to investigate if dietary supplementation prior to ischemic challenge can prevent subsequent damage using two animal models. First, we screened the protective effect against hypoxia–reoxygenation in Drosophila melanogaster and observed that both ethanolic and hydroalcoholic extracts protected flies from the deleterious effects of hypoxia. Second, we confirmed the protective effect of S. ramosissima ethanolic extract against brain ischemia using the transient middle cerebral artery occlusion mice model. Four weeks of oral supplementation with the ethanolic extract before artery occlusion reduced infarct volume and lowered the plasma levels of the DNA peroxidant product 8-hydroxydeoxyguanosine. Phytochemical profiling of S. ramosissima ethanolic extract revealed 50 compounds. Thus, it represents a valuable source of bioactive compounds that show promising disease-modifying activities and could be further developed as an effective food supplement for the prevention or treatment of neurovascular disorders.PE-0527-2019 Programa Operativo FEDERConsejería de Transformación Económica, Industria, Conocimiento y Universidades[PY20_01351]ISCIII, grant number [CD21/00148