The Bioactivity of a Hydroxytyrosol-Enriched Extract Originated after Direct Hydrolysis of Olive Leaves from Greek Cultivars.

Nowadays, olive leaf polyphenols have been at the center of scientific interest due to their beneficial effects on human health. The most abundant polyphenol in olive leaves is oleuropein. The biological properties of oleuropein are mainly due to the hydroxytyrosol moiety, a drastic catechol group, whose biological activity has been mentioned many times in the literature. Hence, in recent years, many nutritional supplements, food products, and cosmetics enriched in hydroxytyrosol have been developed and marketed, with unexpectedly positive results. However, the concentration levels of hydroxytyrosol in olive leaves are low, as it depends on several agricultural factors. In this study, a rapid and easy methodology for the production of hydroxytyrosol-enriched extracts from olive leaves was described. The proposed method is based on the direct acidic hydrolysis of olive leaves, where the extraction procedure and the hydrolysis of oleuropein are carried out in one step. Furthermore, we tested the in vitro bioactivity of this extract using cell-free and cell-based methods, evaluating its antioxidant and DNA-protective properties. Our results showed that the hydroxytyrosol-enriched extract produced after direct hydrolysis of olive leaves exerted significant in vitro antioxidant and geno-protective activity, and potentially these extracts could have various applications in the pharmaceutical, food, and cosmetic industries.

Development of a Holistic In Vitro Cell-Free Approach to Determine the Redox Bioactivity of Agricultural Products.

In recent years, there has been a strong consumer demand for food products that provide nutritional benefits to human health. Therefore, the assessment of the biological activity is considered as an important parameter for the promotion of high-quality food products. Herein, we introduce a novel methodology comprising a complete set of in vitro cell-free screening techniques for the evaluation of the bioactivity of various food products on the basis of their antioxidant capacity. These assays examine the free radical scavenging activities, the reducing properties, and the protective ability against oxidative damage to biomolecules. The adoption of the proposed battery of antioxidant assays is anticipated to contribute to the holistic characterization of the bioactivity of the food product under examination. Consumer motivations and expectations with respect to nutritious food products with bio-functional properties drive the global food market toward food certification. Therefore, the development and application of scientific methodologies that examine the quality characteristics of food products could increase consumers' trust and promote their beneficial properties for human health.

Redox Biomarkers Assessment after Oral Administration of Wine Extract and Grape Stem Extract in Rats and Mice.

Wine and by-products of the winemaking process, such as grape stems, are rich in bioactive polyphenolic compounds that might be beneficial for animal and human health. In recent years, the administration of dietary polyphenols with strong antioxidant and cytoprotective properties has constituted an emerging line of research interest toward disease prevention. However, in scientific literature, only a limited number of studies have investigated the safety and the toxicological risks of polyphenolic compounds in vivo. Based on the above, the purpose of the present study was two-fold: first, to examine the effects of oral administration of a grape stem extract, derived from the Greek red wine Mavrodaphne, on mice redox biomarkers; and second, to investigate the biological effects of oral administration of a wine extract, derived from the emblematic Greek red wine Xinomavro, on rats. Toward this purpose, body weight, growth rate, hematological, biochemical, and histopathological parameters, as well as a panel of redox biomarkers, were examined. According to our results, the administration of Mavrodaphne grape stem extract in mice induced alterations in redox homeostasis, preventing mice from the adverse effects of lipid peroxidation. Contrariwise, the administration of Xinomavro wine extract induced both beneficial and harmful outcomes on rat redox status determined by the examined tissue. Collectively, our study reports that the Mavrodaphne grape stem extract, a serious pollutant when disposed in environmental matrices, is an important source of bioactive polyphenolic compounds that could protect from oxidative damage and improve animal and human health. Finally, the Xinomavro wine extract exerts tissue-specific changes in redox balance, which are indicative of the complexity that characterizes the biological systems.

Assessment of Antioxidant and Antimutagenic Properties of Red and White Wine Extracts In Vitro.

Wine is an alcoholic beverage of complex composition obtained through the fermentation of grape must. The consumption of wine has already been associated with a multitude of beneficial effects due to its high polyphenolic content. In this study, four Greek emblematic wines from two red (i.e., Xinomavro and Agiorgitiko) and two white (i.e., Assyrtiko and Malagouzia) varieties were analyzed for the estimation of their antioxidant profiles. To address this question, we assessed their ability to scavenge both synthetic and endogenous free radicals, such as DPPH•, ABTS+•, OH•, O2-, their potential reducing power, and their antimutagenic and antigenotoxic properties. All varieties exhibited potent antioxidant activity, as indicated by the results of methods above, with the red wines appearing more effective than the white ones regarding antioxidant capacity. Our small-scale study is the first to reveal that these wine varieties may have the ability to scavenge the most reactive endogenous radicals. In the future, this finding must be accompanied by larger studies to fill a knowledge gap in the scientific literature concerning a holistic approach of the in vitro antioxidant action of plant polyphenolic compounds. Conclusively, we believe that wines possess high bioactivity that allow them to settle in the industry of food additives and medicinal products.

A novel combined bioactivity / chemoactivity holistic approach for the evaluation of dietary supplements.

There is increasing evidence that the excessive generation of free radicals in the human body plays a major role in the pathophysiology and development of various diseases, closely associated with oxidative damage. In this frame, the consumption of antioxidant nutrients through food or dietary supplements may prevent from the harmful effects of free radicals on human cells. This work proposes a holistic approach consisting of distinct methodologies, suitable to evaluate the antioxidant and chemoprotective activity of three novel dietary supplements, each one containing active substances with complementary properties. In the first step, this approach includes in vitro studies to evaluate the antioxidant activity of the dietary supplements by measuring the parameters of free radical scavenging capacity, of reducing power activity, as well as, their ability to protect biomolecules from oxidation. Furthermore, the evaluation of their antimutagenic and antigenotoxic effects is also presented. SubsequentlySub, the specific effects of the dietary supplements were examined in three cancer cell lines (HepG2, HeLa, MKN45), by measuring redox biomarkers such as glutathione, reactive oxygen species and thiobarbituric acid reactive substances, using flow cytometry and spectrophotometry. Our results indicate that all the dietary supplements exhibit high antioxidant, antimutagenic, antigenotoxic and lipid protective activity. The most prominent result is their capability to induce oxidative damage on cancer cells via the critical decrease of the levels of their intracellular glutathione, as well as the increase of ROS and lipid peroxidation levels after the administration of non-cytotoxic concentrations. We suggest that the proposed methodology could constitute a valuable tool for the characterization of dietary supplements based on their chemical and functional properties.

Olive oil with high polyphenolic content induces both beneficial and harmful alterations on rat redox status depending on the tissue.

Olive oil (OO) possesses a predominant role in the diet of Mediterranean countries. According to a health claim approved by the European Food Safety Authority, OO protects against oxidative stress­induced lipid peroxidation in human blood, when it contains at least 5 mg of hydroxytyrosol and its derivatives per 20 g. However, studies regarding the effects of a total OO biophenols on redox status in vivo are scarce and either observational and do not provide a holistic picture of their action in tissues. Following a series of in vitro screening tests an OO containing biophenols at 800 mg/kg of OO was administered for 14 days to male Wistar rats at a dose corresponding to 20 g OO/per day to humans. Our results showed that OO reinforced the antioxidant profile of blood, brain, muscle and small intestine, it induced oxidative stress in spleen, pancreas, liver and heart, whereas no distinct effects were observed in lung, colon and kidney. The seemingly negative effects of OO follow the recently formulated idea in toxicology, namely the real life exposure scenario. This study reports that OO, although considered a nutritional source rich in antioxidants, it exerts a tissues specific action when administered in vivo.

Οlive tree blossom polyphenolic extracts exert antioxidant and antimutagenic activities in vitro and in various cell lines.

Olive oil has held a prominent place in the Mediterranean diet since ancient times due to its beneficial effects on human health thus, becoming the subject of great scientific interest. Although numerous studies have examined the biological action of olive and olive oil extracts, the literature lacks studies investigating the putative antioxidant capacity of olive tree flower extracts. Given that olive tree flowers are actually by-products of the olive oil production process with high waste burden for the environment, it becomes evident that their exploitation could increase their added value. Therefore, in this study the potential antioxidant action of four olive flower extracts was investigated. All the extracts exerted potent antioxidant activity as indicated using the DPPH• and ABTS•+ assays, as well as antigenotoxic and antimutagenic properties, identified by the results of the plasmid relaxation assay and the Ames test, respectively. Furthermore, the extracts also improved redox status of four cell lines (i.e., EA.hy926, C2C12, HeLa, and HepG2) enhancing reduced glutathione and reducing reactive oxygen species levels using flow cytometry. Taking into account that during olive tree cultivation a considerable amount of olive flowers is generated, the waste burden is high and the management is difficult. Given the optimistic findings of the present study, we believe that the flower-derived extracts may have high added value since they could be used as antioxidants or as foodstuff, food additives and functional food constituents.