Application of Green Technology to Extract Clean and Safe Bioactive Compounds from Tetradesmus obliquus Biomass Grown in Poultry Wastewater.

Microalgae are capable of assimilating nutrients from wastewater (WW), producing clean water and biomass rich in bioactive compounds that need to be recovered from inside the microalgal cell. This work investigated subcritical water (SW) extraction to collect high-value compounds from the microalga Tetradesmus obliquus after treating poultry WW. The treatment efficiency was evaluated in terms of total Kjeldahl nitrogen (TKN), phosphate, chemical oxygen demand (COD) and metals. T. obliquus was able to remove 77% TKN, 50% phosphate, 84% COD, and metals (48-89%) within legislation values. SW extraction was performed at 170 °C and 30 bar for 10 min. SW allowed the extraction of total phenols (1.073 mg GAE/mL extract) and total flavonoids (0.111 mg CAT/mL extract) with high antioxidant activity (IC50 value, 7.18 µg/mL). The microalga was shown to be a source of organic compounds of commercial value (e.g., squalene). Finally, the SW conditions allowed the removal of pathogens and metals in the extracts and residues to values in accordance with legislation, assuring their safety for feed or agriculture applications.

Comparative Study of the Essential Oil and Hydrosol Composition of Sweet Wormwood (Artemisia annua L.) from Serbia.

The most abundant volatile compounds of sweet wormwood (Artemisia annua L.) essential oil were artemisia ketone (25.4 %) and trans-caryophyllene (10.2 %), followed by 1,8-cineole, camphor, germacrene D and ß-selinene. The major volatile compounds in the hydrosol were camphor (25.1 %), 1,8-cineole (20.5 %) and artemisia ketone (10.7 %), followed by trans-pinocarveol and yomogi alcohol. Tested essential oil was rich in oxygenated monoterpenes and sesquiterpene hydrocarbons, while the former were identified as the major class of volatile compounds in the hydrosol, due to higher water solubility. Classification of all sweet wormwood chemotypes, according to essential oil composition, in available literature (17 studies and 61 accessions) could be done according to four chemotypes: artemisia ketone+artemisia alcohol (most abundant), artemisia ketone, camphor and nonspecific chemotype. According to this classification, essential oil of sweet wormwood from this study belongs to artemisia ketone (content varied between 22.1 and 55.8 %). Bearing in mind that hydrosols are a by-product of industrial production of essential oils, and the fact that sweet wormwood hydrosol has high contents of camphor, 1,8-cineole and artemisia ketone, there is a great potential for the use of this aromatic plant primary processing waste product as a water replacement in cosmetic industry, beverages flavoring, for food preservation, as well as in post-harvest pre-storage treatments in organic agriculture.

Supercritical CO2 Extract from Microalga Tetradesmus obliquus: The Effect of High-Pressure Pre-Treatment.

High-pressure pre-treatment followed by supercritical carbon dioxide (ScCO2) extraction (300 bar, 40 °C) was applied for the attainment of the lipophilic fraction of microalga Tetradesmus obliquus. The chemical profile of supercritical extracts of T. obliquus was analyzed by ultra-high-performance liquid chromatography-high-resolution mass spectrometry with electrospray ionization (UHPLC-ESI-HRMS). Moreover, the impact of ScCO2 on the microbiological and metal profile of the biomass was monitored. The application of the pre-treatment increased the extraction yield approximately three-fold compared to the control. In the obtained extracts (control and pre-treated extracts), the identified components belonged to triacylglyceroles, fatty acid derivatives, diacylglycerophosphocholines and diacylglycerophosphoserines, pigments, terpenes, and steroids. Triacylglycerols (65%) were the most dominant group of compounds in the control extract. The pre-treatment decreased the percentage of triacylglycerols to 2%, while the abundance of fatty acid derivatives was significantly increased (82%). In addition, the pre-treatment led to an increase in the percentages of carotenoids, terpenoids, and steroids. Furthermore, it was determined that ScCO2 extraction reduced the number of microorganisms in the biomass. Considering its microbiological and metal profiles, the biomass after ScCO2 can potentially be used as a safe and important source of organic compounds.

Evaluation of In Vitro Cytotoxic Potential of Avarol towards Human Cancer Cell Lines and In Vivo Antitumor Activity in Solid Tumor Models.

The goal of this study was to determine the activity in vitro and in vivo of avarol, a sesquiterpene hydroquinone originating from the Dysidea avara sponge from the south Adriatic Sea, against different cancer cell lines and two types of mouse carcinoma. To investigate the in vitro cytotoxicity, a human cervix adenocarcinoma cell line (HeLa), human colon adenocarcinoma (LS174), human non-small-cell lung carcinoma (A549), and a normal human fetal lung fibroblast cell line (MRC-5) were used. The in vivo antitumor activity was investigated against two transplantable mouse tumors, the Ehrlich carcinoma (EC) and cervical cancer (CC-5). The effect of avarol on cancer cell survival, which was determined by the microculture tetrazolium test, confirmed a significant in vitro potency of avarol against the investigated cell lines, without selectivity towards MRC-5. The highest cytotoxicity was exhibited against HeLa cancer cells (10.22 ± 0.28 µg/mL). Moreover, potent antitumor activity against two tumor models was determined, as the intraperitoneal administration of avarol at a dose of 50 mg/kg resulted in a significant inhibition of tumor growth in mice. After three administrations of avarol, a 29% inhibition of the EC growth was achieved, while in the case of CC-5, a 36% inhibition of the tumor growth was achieved after the second administration of avarol. Therefore, the results indicate that this marine sesquiterpenoid hydroquinone could be a promising bioactive compound in the development of new anticancer medicine.

Supercritical Carbon Dioxide Extraction of Allium ursinum: Impact of Temperature and Pressure on the Extracts Chemical Profile.

The aim of this study was to extract Allium ursinum L. for the first time by supercritical carbon dioxide (SC-CO2 ) as green sustainable method. The impact of temperature in the range from 40 to 60 °C and pressure between 150 and 400 bar on the quality of the obtained extracts and efficiency of the extraction was investigated. The highest extraction yield (3.43 %) was achieved by applying the extraction conditions of 400 bar and 60 °C. The analysis of the extracts was performed by gas chromatography and mass spectrometry (GC/MS). The most dominant sulfur-containing constituent of the extracts was allyl methyl trisulfide with the highest abundance at 350 bar and 50 °C. In addition, the presence of other pharmacologically potent sulfur compounds was recorded including S-methyl methanethiosulfinate, diallyl trisulfide, S-methyl methylthiosulfonate, and dimethyl trisulfide. Multivariate data analysis tool was utilized to investigate distributions of the identified compounds among the extracts obtained under various extraction conditions and yields. It was determined that the SC-CO2 extraction can by efficiently used for A. ursinum.

Optimization of MAE for the Separation of Nicotine and Phenolics from Tobacco Waste by Using the Response Surface Methodology Approach.

This study intends to valorize by-products of the industrial processing of tobacco to obtain nicotine and phenolics as value-added compounds. Three influential parameters of the microwave-assisted extraction-MAE (temperature, treatment time, and solvent/solid ratio) were studied for the optimization of the extraction protocol for tobacco leaves and three types of waste-scrap, dust, and midrib, respectively. Nicotine was the dominant bioactive compound in all extracts, ranging from 1.512 to 5.480% in leaves, 1.886 to 3.709% in scrap, 2.628 to 4.840% dust, and 0.867 to 1.783% in midrib extracts. Five phenolic compounds were identified and quantified, predominated by chlorogenic acid and rutin. Additionally, total phenol content and antioxidant activity were determined using spectrophotometric assays. Optimization was performed in two aspects: to obtain a maximum extraction yield with minimum nicotine content and to obtain a maximum extraction yield with maximum nicotine content. These findings demonstrate that tobacco waste is a valuable source of bioactive compounds and MAE can be a promising alternative technique to obtain extracts rich in targeted bioactive compounds, especially nicotine.

Valorization of Yarrow (Achillea millefolium L.) By-Product through Application of Subcritical Water Extraction.

In the present study, valorization of yarrow (Achillea millefolium) by-product from the filter tea industry was investigated through the application of subcritical water for the extraction of bioactive compounds. The influence of different process parameters (temperature 120-200 °C, extraction time 10-30 min, and HCl concentration in extraction solvent 0-1.5%) on extract quality in terms of content of bioactive compounds and antioxidant activity was investigated. Optimal conditions of the extraction process (temperature 198 °C, extraction time 16.5 min, and without acidifer) were determined and, when applied, the most efficient exploitation of by-products is achieved, that is, attainment of extracts rich in total phenols and flavonoids and high antioxidant activity. In addition, by applying the high performance liquid chromatographic analysis, the content of chlorogenic acid was determined as well as the hydroxymethylfurfural content in obtained extracts. The results demonstrated that subcritical water can be successfully used for utilization of yarrow by-products for obtaining extracts rich in antioxidants.

Comparative Study of Subcritical Water and Microwave-Assisted Extraction Techniques Impact on the Phenolic Compounds and 5-Hydroxymethylfurfural Content in Pomegranate Peel.

Two environmentally friendly innovative extraction techniques - subcritical water (SWE) and microwave-assisted extraction (MAE) were applied for the extraction of phenolics from pomegranate peel. The impact of process conditions (SWE: temperature 100-220 °C, extraction time 5-30 min; MAE: solvent water and 50% ethanol, irradiation power 470 and 800 W) on the quality of extracts in terms of the content of total phenolics, total flavonoids, major phenolic constituents (gallic acid, ellagic acid, punicalin, punicalagin), as well as 5-hydroxymethylfurfural(HMF) amount was investigated. For SWE, temperature of 130 °C and 20 min extraction time were found optimal for obtaining high content of bioactive compounds and minimizing the yield of HMF. During MAE, phenolic compounds were effectively extracted by using lower microwave power and 50% ethanol. Comparing two techniques, MAE is more efficient than SWE for the extraction of phenolics from pomegranate peel while obtaining a HMF-free extracts.

Aronia Berry Processing by Spray Drying: From Byproduct to High Quality Functional Powder.

The main aim of this study is to analyze the solid-liquid extraction followed by spray drying as a technological pathway for utilization of aronia fruit dust, a byproduct of filter tea factory. In the current study, ultrasound-assisted extraction was applied for the production of aronia liquid feed and maltodextrin was used as a carrier and encapsulating agent. In spray drying, the influence of inlet temperature and maltodextrin type and mass fraction on process efficiency and powder properties were observed. The physical and chemical properties of the obtained powders were characterized. It was determined that the powder produced using inlet temperature 140 °C and 40% maltodextrin with dextrose equivalent (DE) 19.7 had the most desirable characteristics. It was observed that the increase in maltodextrin mass fraction decreases the powder moisture content, hygroscopicity and the content of bioactive compounds, but increases water solubility index and particle size. The increase in dextrose equivalent of maltodextrin increases the powder hygroscopicity and water solubility index, while the increase of inlet temperature causes a decrease in moisture content of aronia powders.

Recovery of Antioxidant Compounds from Aronia Filter Tea Factory by -Product: Novel Versus Conventional Extraction Approaches.

Black chokeberry (Aronia melanocarpa L.) by-product from filter tea factory underwent subcritical water extraction (SWE) in order to recover polyphenolics and determine its antioxidant potential. In the current study Box-Behnken design was applied for optimization. Independent variables used in experimental design were temperature (T, 120-200 °C), extraction time (t, 15-35 min) and hydrochloride concentration (c, 0-1.5%). Experimental results were fitted to a second-order polynomial model where multiple regression analysis and analysis of variance were used to determine fitness of the model and optimal conditions. The optimized SWE conditions for maximum responses of total phenols (TP), total flavonoids (TF) and monomeric anthocyanins (MA) contents, and minimum response of IC50 were temperature of 120.4 °C, extraction time of 15.2 min and absence of acidifier. The predicted values of TP, TF, MA and IC50 at these conditions were: 32.8863 mg GAE/g, 23.5164 mg CE/g, 0.5124 mg C3G/g and 0.0055 mg/mL, respectively.

Supercritical CO2 Extraction of Lavandula angustifolia Mill. Flowers: Optimisation of Oxygenated Monoterpenes, Coumarin and Herniarin Content.

INTRODUCTION: Lavandula angustifolia is good source of oxygenated monoterpenes containing coumarins as well, which are all soluble in supercritical CO2 (SC-CO2 ). OBJECTIVE: The study objective is to investigate SC-CO2 extraction parameters on: the total yield; GC-MS profile of the extracts; relative content of oxygenated monoterpenes; the amount of coumarin and herniarin; and to determine optimal SC-CO2 extraction conditions by response surface methodology (RSM). METHODOLOGY: SC-CO2 extraction was performed under different pressure, temperature and CO2 flow rate determined by Box-Behnken design (BBD). The sample mass and the extraction time were kept constant. The chemical profiles and relative content of oxygenated monoterpenes (as coumarin equivalents, CE) were determined by GC-MS. Coumarin and herniarin concentrations were dosed by HPLC. RESULTS: SC-CO2 extracts contained linalool (57.4-217.9 mg CE/100 g), camphor (10.6-154.4 mg CE/100 g), borneol (6.2-99.9 mg CE/100 g), 1,8-cineole (5.0-70.4 mg CE/100 g), linalyl acetate (86.1-267.9 mg CE/100 g), coumarin (0.95-18.16 mg/100 g), and herniarin (0.95-13.63 mg/100 g). The interaction between the pressure and CO2 flow rate as well as between the temperature and CO2 flow rate showed statistically significant influence on the extraction yield. Applying BBD, the optimum extraction conditions for higher monoterpenes and lower coumarin content were at 10 MPa, 41°C and CO2 flow rate 2.3 kg/h, and at 30 MPa, 50°C and CO2 flow rate 3 kg/h for higher monoterpenes and coumarin content. CONCLUSION: SC-CO2 extraction is a viable technique for obtaining lavender extracts with desirable flavour components. The second-order model based on BBD predicts the results for SC-CO2 extraction quite satisfactorily. Copyright © 2017 John Wiley & Sons, Ltd.

Optimization of microwave-assisted extraction (MAE) of coriander phenolic antioxidants - response surface methodology approach.

BACKGROUND: Microwave-assisted extraction (MAE) of polyphenols from coriander seeds was optimized by simultaneous maximization of total phenolic (TP) and total flavonoid (TF) yields, as well as maximized antioxidant activity determined by 1,1-diphenyl-2-picrylhydrazyl and reducing power assays. Box-Behnken experimental design with response surface methodology (RSM) was used for optimization of MAE. Extraction time (X1 , 15-35 min), ethanol concentration (X2 , 50-90% w/w) and irradiation power (X3 , 400-800 W) were investigated as independent variables. Experimentally obtained values of investigated responses were fitted to a second-order polynomial model, and multiple regression analysis and analysis of variance were used to determine fitness of the model and optimal conditions. RESULTS: The optimal MAE conditions for simultaneous maximization of polyphenol yield and increased antioxidant activity were an extraction time of 19 min, an ethanol concentration of 63% and an irradiation power of 570 W, while predicted values of TP, TF, IC50 and EC50 at optimal MAE conditions were 311.23 mg gallic acid equivalent per 100 g dry weight (DW), 213.66 mg catechin equivalent per 100 g DW, 0.0315 mg mL(-1) and 0.1311 mg mL(-1) respectively. CONCLUSION: RSM was successfully used for multi-response optimization of coriander seed polyphenols. Comparison of optimized MAE with conventional extraction techniques confirmed that MAE provides significantly higher polyphenol yields and extracts with increased antioxidant activity. © 2016 Society of Chemical Industry.

Acetylcholinesterase inhibitory, antioxidant and phytochemical properties of selected medicinal plants of the Lamiaceae family.

The present study aimed to evaluate acetylcholinesterase (AChE) inhibitory and antioxidant activities of Lamiaceae medicinal plants growing wild in Croatia. Using Ellman's colorimetric assay all tested ethanolic extracts and their hydroxycinnamic acid constituents demonstrated in vitro AChE inhibitory properties in a dose dependent manner. The extracts of Mentha x piperita, M. longifolia, Salvia officinalis, Satureja montana, Teucrium arduini, T. chamaedrys, T. montanum, T. polium and Thymus vulgaris at 1 mg/mL showed strong inhibitory activity against AChE. The antioxidant potential of the investigated Lamiaceae species was assessed by DPPH• scavenging activity and total antioxidant capacity assays, in comparison with hydroxycinnamic acids and trolox. The extracts differed greatly in their total hydroxycinnamic derivatives content, determined spectrophotometrically. Rosmarinic acid was found to be the predominant constituent in most of the investigated medicinal plants (by RP-HPLC) and had a substantial influence on their AChE inhibitory and antioxidant properties, with the exception of Teucrium species. These findings indicate that Lamiaceae species are a rich source of various natural AChE inhibitors and antioxidants that could be useful in the prevention and treatment of Alzheimer's and other related diseases.

A new green approach for Lavandula stoechas aroma recovery and stabilization coupling supercritical CO2 and natural deep eutectic solvents.

This work investigated a green approach to obtain and stabilize Lavandula stoechas L. volatile organic compounds with sensory aroma characteristics by using alternative solvents, namely supercritical carbon dioxide (scCO2) and deep eutectic solvents (DES). The CO2 extracts were dispersed in different DES mixtures (betaine:ethylene glycol (1:3), betaine:glycerol (1:2), and glycerol:glucose (4:1)) and their stability was monitored during 6 months of storage at room temperature by monitoring the headspace (HS) profile. The CO2 extract was used as the control. It was initially determined that there was a dominant presence of oxygenated monoterpenes (67.33-77.50%) in the extracts. During storage, significant changes occurred in the samples' HS, such as the decrease in terpene hydrocarbons which also affected the presence of oxygenated terpenes, which increased in certain cases. Moreover, the highest formation of new components was recorded in the control which could be an indicator of decreased stability. The DESs-CO2 were more stable than the CO2 control and among them, betaine:ethylene glycol stood out as the most adequate systems for maintaining the stability of L. stoechas HS components. For the visual estimation of similarities and dissimilarities among the samples, chemometric pattern recognition approaches were applied including the hierarchical cluster analysis, principal component analysis, and sum of ranking differences.

Innovative Strategy for Aroma Stabilization Using Green Solvents: Supercritical CO2 Extracts of Satureja montana Dispersed in Deep Eutectic Solvents.

The aim of this work was to establish the potential of natural deep eutectic solvents (NADES) for the stabilization of aroma volatile organic compounds from a natural source. Satureja montana was used as a source of volatile components, as it is rich in terpenes of great commercial and biological importance, such as carvacrol, thymol, and thymoquinone, among others. Supercritical CO2 was used to extract the lipophilic fraction of S. montana, which was further directly dispersed in NADES. The stabilizing capacity of seven different NADES based on betaine and glycerol was analyzed. The stability of the components in NADES was monitored by analyzing the headspace profile during 6 months of storage at room temperature. The changes in the headspace profile over time were analyzed by using different statistical and chemometric tools and the Wilcoxon matched pair test. It was determined that alterations over time occurred such as degradation and oxidation, and they were the most prominent in the control. In addition, the indicator of decreased stability of the control was the formation of the new compounds that could compromise the quality of the product. In the stabilized NADES samples, the changes were significantly less prominent, indicating that the NADES had a stabilizing effect on the volatile compounds. According to Wilcoxon matched pair test, the most efficient stability was achieved by using betaine/ethylene glycol, glycerol/glucose, and betaine/sorbitol/water. Therefore, by applying two green solvents, a sustainable approach for obtaining pure and high-quality S. montana extracts with extended stability at room temperature was established.

Towards a Greener Approach for Biomass Valorization: Integration of Supercritical Fluid and Deep Eutectic Solvents.

A green and sustainable procedure for obtaining Lavandula stoechas extracts with antioxidant and antimicrobial properties was investigated. Green solvents, supercritical CO2, and natural deep eutectic solvents (NADES) together with ultrasound-assisted extraction were used for the sequential extraction of terpene and polyphenols fractions. After the CO2 extraction of the terpene fraction, the residue material was used in an extraction with different NADES (betaine-ethylene glycol (Bet:EG), betaine-glycerol (Bet:Gly), and glycerol-glucose (Gly:Glu)), intensified with an ultrasound-assisted method (at 30 and 60 °C). In the CO2 extract, the major group of components belonged to oxygenated monoterpenes, while the highest polyphenol content with the dominant rutin (438.93 ± 4.60 µg/mL) was determined in Bet:EG extracts (60 °C). Bet:EG extracts also exhibited the most potent antioxidant activity according to DPPH, ABTS, and FRAP assays. Moreover, Bet:EG extracts showed significant inhibitory activity against Gram-positive and Gram-negative bacteria, with minimum inhibitory activity of 0.781-3.125 and 1.563-6.250 mg·mL-1, respectively. By comparing the polyphenolic content and antioxidant and antimicrobial activities of Bet:EG extracts with extracts obtained with conventional solvents (water and ethanol), the superiority of NADES was determined. The established environmentally friendly procedure unifies the requirements of green and sustainable development and modern pharmacognosy because it combines the use of safe alternative solvents, the absence of solvent waste generation, more rational use of resources, and the attainment of safe and quality extracts.

Recovery of Polyphenols from Rosehip Seed Waste Using Natural Deep Eutectic Solvents and Ultrasonic Waves Simultaneously.

Rosehips are processed and consumed in numerous forms, such as juice, wine, herbal tea, yogurt, preserved fruit, and canned products. The seeds share in fruit is 30-35% and they have recently been recognized as an important source of oil rich in unsaturated fatty acids. However, after defatting, seed waste may still contain some polar polyphenolic compounds, which have been scarcely investigated. The aim of this study was to examine the potential of the defatted seed waste as a source of polyphenols. For the defatting process, supercritical carbon dioxide extraction at 300 bar and 40 °C was applied. The capacity of eight different natural deep eutectic solvents (NADES) for the recovery of phenolics from defatted rosehip seed powder (dRSP) was examined. In the extracts obtained with ultrasound-assisted NADES extraction, twenty-one phenolic compounds were identified with LC-MS/MS, among which the most abundant were quinic acid (22.43 × 103 µg/g dRSP) and catechin (571.93 µg/g dRSP). Ternary NADES formulations based on lactic acid proved to be superior. Potential correlations between identified chemical compounds, solvent polarity and viscosity, as well as the compound distributions across studied solvent combinations in PCA hyperspace, were also investigated. PCA demonstrated that more polar NADES mixtures showed improved extraction potential. The established environmentally friendly process represents an approach of transforming rosehip seed waste into value-added products with the potential to be applied in the food industry and to contribute to sustainable production.

Effects of different extraction methods and conditions on the phenolic composition of mate tea extracts.

A simple and rapid HPLC method for determination of chlorogenic acid (5-O-caffeoylquinic acid) in mate tea extracts was developed and validated. The chromatography used isocratic elution with a mobile phase of aqueous 1.5% acetic acid-methanol (85:15, v/v). The flow rate was 0.8 mL/min and detection by UV at 325 nm. The method showed good selectivity, accuracy, repeatability and robustness, with detection limit of 0.26 mg/L and recovery of 97.76%. The developed method was applied for the determination of chlorogenic acid in mate tea extracts obtained by ethanol extraction and liquid carbon dioxide extraction with ethanol as co-solvent. Different ethanol concentrations were used (40, 50 and 60%, v/v) and liquid CO2 extraction was performed at different pressures (50 and 100 bar) and constant temperature (27 ± 1 °C). Significant influence of extraction methods, conditions and solvent polarity on chlorogenic acid content, antioxidant activity and total phenolic and flavonoid content of mate tea extracts was established. The most efficient extraction solvent was liquid CO2 with aqueous ethanol (40%) as co-solvent using an extraction pressure of 100 bar.

The Application of Ultrasonic Waves and Microwaves to Improve Antihyperglycaemic and Antimicrobial Activities of Marrubium vulgare Extracts.

Having scarce information about ultrasound assisted extraction (UAE) and microwave assisted extraction (MAE) of white horehound (Marrubium vulgare L.), the idea has emerged to determine the optimal process parameters for the maximization of polyphenols and to compare the efficiency of these green extraction technologies. The optimal UAE parameters are temperature of 73.6 °C, extraction time of 40 min and ultrasound power of 30.3 W/L, while the optimal MAE parameters are 63.8% ethanol, extraction time of 15 min and microwave power of 422 W. Extract obtained at optimal UAE parameters shows the highest antihyperglycemic activity (α-amylase inhibition: 50.63% and α-glucosidase inhibition: 48.67%), which can potentially be explained by the presence of chlorogenic acid and quercetin, which were not identified in the macerates. The most sensitive bacterial strain to optimal ultrasonic extract is Bacillus cereus, whereas the most sensitive fungal strain is Saccharomyces cerevisiae.

The Effects of Resveratrol-Rich Extracts of Vitis vinifera Pruning Waste on HeLa, MCF-7 and MRC-5 Cells: Apoptosis, Autophagia and Necrosis Interplay.

Resveratrol is a well-studied plant-derived molecule in cancer biology, with a plethora of documented in vitro effects. However, its low bioavailability and toxicity risk hamper its wider use. In this study, vine shoots after pruning were used as a source of resveratrol (RSV). The activity of subcritical water extract (SWE) and dry extract (DE) is examined on three cell lines: HeLa, MCF-7 and MRC-5. The cytotoxic effect is assessed by the MTT test and EB/AO staining, levels of apoptosis are determined by Annexin V assay, autophagia by ULK-1 expression using Western blot and NF-kB activation by p65 ELISA. Our results show that both resveratrol-rich extracts (DE, SWE) have a preferential cytotoxic effect on malignant cell lines (HeLa, MCF-7), and low cytotoxicity on non-malignant cells in culture (MRC-5). Further experiments indicate that the investigated malignant cells undergo different cell death pathways. MCF-7 cells died preferentially by apoptosis, while the HeLa cells died most likely by necrosis (possibly ferroptosis). Protective autophagia is diminished upon treatment with DE in both HeLa and MCF-7 cells, while SWE does not influence the level of autophagia. The extracts are effective even at low concentrations (below IC50) in the activation of NF-kB (p65 translocation).