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The utilization of biopolymers gained recent attention worldwide due to their effective role in producing eco-friendly and economical products. Mucilage from Opuntia ficus-indica (OFI, and other succulent plants) has the capacity to absorb huge amounts of water, forming a viscous colloid with interesting rheological properties. It also had the potential ability for use as food additives or food technological products. We, therefore, aimed at extracting and characterizing pure liquid mucilage from the cladodes of OFI (Egyptian variety) using multidimensional approaches including HPLC-RID, FTIR, XRD, NMR, DSC, and TGA assays. The chemical composition, total polyphenols, total flavonoids, total flavonols, and antioxidant capacity by DPPH and ABTS assays were also measured. HPLC-RID analysis showed that the mucilage’s sugars are composed of glucose, xylose, rhamnose, galacturonic acid, arabinose, galactose, and fucose with total sugars of 0.375 mg/mL, which were subsequently confirmed by FTIR and NMR results. The specific signals of these sugars were also observed in 13C and 1H NMR spectra and their chemical fingerprint was obtained by FTIR. XRD patterns showed that mucilage has high calcium content, and the glass transition temperature was observed at 85.9 °C. Meanwhile, total phenolic content, flavonoids, and flavonols were about 7.96 mg GAE/g FW, 3.61 mg QE/g FW, and 1.47 mg QE/g FW, respectively. The antioxidant capacity of mucilage was around 26.15 and 22.5 µmol TE/g FW for DPPH and ABTS methods, respectively. It can be concluded that OFI cladode mucilage showed promising properties that would improve and open new opportunities and trends in the food, pharmaceutical, cosmetic, and other industries. The use of pure liquid mucilage could also be economically profitable due to its low cost, availability, and effectiveness for many edible applications.

To explore the effect of pectolytic enzyme application on polysaccharide size and colloidal interactions, rosé wines were prepared from pressed Shiraz grape juice, and red wines were made from the same juice by the addition of skins and seeds from fresh pressings. A pectolytic enzyme preparation containing primarily polygalacturonase and side-activities of arabinase and pectin lyase was used. Enzyme treatment enhanced the contribution of high molecular weight (≈ 200 kDa) polysaccharides rich in mannose and glucose (mannoproteins) and removed arabinose-rich polysaccharides of intermediate (≈ 40 kDa) size. Enzyme application reduced the molecular weight average of a class of smaller polysaccharides of approximately 6 kDa (rhamnogalacturonan II). All wines had similar particle sizes, but enzyme treatment markedly reduced particle concentration in wines, particularly in rosé wines. Wine polysaccharides were purified and when reconstituted in model wine, showed reduced particle concentration in response to enzyme treatment. Aggregation of polysaccharide in the presence of seed tannin was also markedly reduced by enzyme treatment. The results indicated that changes in structure introduced by the enzyme affected wine colloidal properties, potentially increasing polysaccharide solubility.

The carrot is one of the most cultivated vegetables in the world. Black or purple carrots contain acylated anthocyanins which are of special interest to the food industry for their stability and nutraceutical characteristics. Anthocyanin-rich fruits and vegetables have gained popularity in the last ten years, due to the health benefits they provide. In this paper, the characterizations of the bioactive compounds and antioxidant capacities of different anthocyanin-containing carrots (a black carrot—BC, and a local purple carrot, the “Polignano” carrot—PC), compared to the commercial orange carrot (OC) (lacking of anthocyanins), are reported. The anthocyanin profiles of the polyphenolic extracts of BC and PC were similar, but differences were observed at quantitative levels. The total anthocyanin content in BC was more than twice that in PC (13.84 ± 0.61 vs. 5.64 ± 0.48 mg K Eq. g−1 DW). Phenolic acids (mostly chlorogenic acid) were also present at high level in anthocyanin-rich carrots compared to OC. High polyphenol content accounted also for a high reducing capacity (evaluated by Folin–Ciocalteu reagent, FCR), and antioxidant capacity (evaluated by TEAC and ORAC assays) which were the highest for BC (FCR value: 16.6 ± 1.1 mg GAE. g−1 DW, TEAC: 76.6 ± 10.6 µmol TE. g−1 DW, ORAC: 159.9 ± 3.3 µmol TE. g−1 DW). All carrot genotypes (mostly OC) were rich in carotenoids (BC 0.14 ± 0.024, PC 0.33 ± 0.038, OC 1.29 ± 0.09 mg. g−1 DW), with predominance of α and β-carotene, in OC, and lutein in BC. PC showed the highest malic acid and sugar (glucose plus fructose) content. In conclusion, while BC is remarkable for nutraceutical features, the local genotype (“Polignano” carrot) is worth considering in genetic biodiversity conservation programme.

Rose-scented geranium (Pelargonium sp.) have been the subject of many studies concerning essential oil, unlike the edible flowers of scent Pelargonium despite the beneficial properties of chemical compounds present in such organ and the fragrances produced. Therefore, we investigated organic compounds, total phenols and the antioxidant activity of the flowers with particular attention to the volatile compounds emitted by flowers of Pelargonium ‘Endsleigh’ at three stages of development. We identified several major phenols and organic compounds that revealed significative difference compared with other Pelargonium flowers, and employing the solid-phase micro extraction (SPME) methodology we determined that the production of volatile compounds decreases during the flower development. In addition, the emission of volatiles is accomplished mainly by the sepals. In conclusion, to use Pelargonium ‘Endsleigh’ flowers, it is important in ensuring that they have not yet opened and that the sepals are in place.

BACKGROUND: Bioactive compounds, mainly polyphenols, present in berries, are thought to be responsible for the health benefits of these fruit. Therefore, it is worthwhile to define the optimal environmental conditions to maximise their polyphenol content. OBJECTIVE: With the aim to define the optimal conditions for berry cultivation in an innovative environment, red raspberry, wild strawberry and blackberry plants were grown in a traditional greenhouse in comparison with two photovoltaic greenhouses with different shading area. METHODS: Hydroalcoholic extracts of ripe berries were evaluated by HPLC analysis, for their anthocyanins, organic acids and sugar contents. Moreover, phenolic content (by the Folin-Ciocalteu assay) and antioxidant activity (by the Trolox equivalent antioxidant capacity-TEAC assay) were assayed on the same berry extracts. RESULTS: Total anthocyanins, phenols content and antioxidant capacity tended to increase in berries grown under shading. The sugars content was, mostly, not negatively influenced by the shading. Conversely, the organic acids content, in some instances, increased along with the shading percentage. CONCLUSIONS: It can be concluded that it is possible to combine the greenhouse production of high-quality berries (with a particular focus on polyphenols, bioactive compounds valuable for human health) with the production of renewable energy, in the context of sustainable agriculture.

This study investigates the effects of tomato puree fortification with several anthocyanin-rich food colorants on bioactive compound content (phenolics, isoprenoids), antioxidant capacity, in vitro biological activities and consumer acceptance. Tomato puree (tp) was added with different anthocyanin extracts from black carrot (Anthocarrot), grape fruit skins (Enocolor), elderberry fruits (Elderberry) or mahaleb cherry fruits (Mahaleb), thus obtaining a 'functional tomato puree' (ftp). The consumer acceptance (colour, flavor, taste, visual appearance) was at high level, except for Mahaleb-added ftp. Compared to the control (tp), the addition of colouring extracts increased significantly the total phenolic content, before pasteurization, in addition to the expected anthocyanin content. However, after pasteurization, mostly Anthocarrot-ftp preserved an increased phenolic (+53%) content, as well as a higher antioxidant capacity (50%), more than the other added-extracts. Consistently, against tp, Anthocarrot-ftp exhibited an increased anti-inflammatory capacity as showed by the reduced expression of vascular cell adhesion molecule (VCAM)-1 in human cultured endothelial cells, under inflammatory conditions.

Artemisia annua L. is a herbaceous plant belonging to the Asteraceae family, known for producing, although at low levels, the sesquiterpene lactone artemisinin (AN), which is highly effective against malaria. In this study, an in vitro regeneration process of A. annua L. using ‘Artemis’ progeny was established and the potential of tissue culture for inducing new variability in terms of AN metabolism of in vitro regenerated plants was investigated. Among the plant growth regulators tested, the cytokinin 6-benzyladenine (BA) at 4.4 μM in combination with the auxin indole-butyric acid (IBA) at 0.35 μM yielded the greatest frequency of shoot induction. The optimal multiplication medium contained BA at 0.9 μM and naphthaleneacetic acid (NAA) at 0.05 μM. Regenerated plants (RPs), after transferring to the greenhouse and subsequently to the field, were analyzed during the growth cycle at different sampling times, showing a peak of AN content 20 days before blossom. Variability among different RPs and sampling times, in terms of AN and its precursors dihydroartemisinic acid (DHAA) and artemisinic acid (AA) was observed. This suggests that adventitious shoot induction could provide a useful strategy to induce variability influencing artemisinin metabolism as a consequence of in vitro manipulation.

[Context]: The inclusion of Pinus taeda hydrolysed lignin (PTHL) in beef diets could improve quality and stability of meat, but effects could vary through the aging period (AP)., [Aim]: The aim was to evaluate the effects of the PTHL inclusion in the diet of finishing beef cattle on meat quality, fatty acid composition and oxidative stability at Days 1, 8, 11 and 15 of aging., [Methods]: Forty Limousin bulls (340 ± 42 kg) were fed ad libitum on a total mixed ration (TMR). The control group received exclusively TMR for 120 days, while the experimental group received the same TMR as the control group but supplemented with PTHL (Oxifenol, I-Green, Padua, Italy; 35 g/day per head at 1–90 days and 70 g/day per head at 91–120 days)., [Key results]: Diet did not influence the chemical composition, pH, cooking loss, Warner–Bratzler shear force and hydroperoxide content. The Warner–Bratzler shear force (P < 0.001) decreased, while lightness (P < 0.01) and hydroperoxides (P < 0.001) increased through the aging period. Thiobarbituric acid reactive substances were similar for both diets at 1 and 15 days. However, thiobarbituric acid reactive substances at 8 and 11 days were higher for control than for the PTHL diet (P = 0.023 for interaction). Protein carbonyls were higher for control than for the PTHL diet at 8 days (P = 0.003 for interaction), but similar for both diets for the other dates. Saturated, monounsaturated and polyunsaturated fatty acids varied through the AP with PTHL diet, while no changes were observed with control diet (P < 0.01 for interactions). At 11 days, the n-6 : n-3 ratio passed from being the minimum value with the PTHL diet to be the maximum with control diet (P < 0.01 for interaction)., [Conclusions]: The effects of PTHL inclusion in bull finishing diets depends on the AP but, generally, may result in beef with meat with beneficial effects on human health., [Implications]: Including PTHL in the diet of finishing bulls can be useful to improve meat quality, favouring the use of natural waste substances deriving from vegetal production.

Morchella pulchella Clowez & Franç. Petit. and Helvella leucopus Pers. are among the most utilized naturally grown wild-edible mushroom species in Eastern Anatolia, Turkey. This study focused on revealing their phenolic profiles using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), antioxidant (hydrogen atom and single electron-transfer mechanisms), and enzyme inhibition (acetylcholinesterase, amylase, glucosidase, and lipase) activities of lyophilized sequential (n-hexane, ethanol, methanol, and water, respectively) extracts and traditional preparations (infusion and decoction) obtained from the fruiting bodies of these mycomaterials. Our chromatographic studies revealed that M. pulchella extracts are dominated by caffeic acid, while H. leucopus extracts contained significant levels of protocatechuic acid as a major biologically active compound (excluding n-hexane fractions). The most active antioxidant and enzyme inhibitory extracts were from ethanol (or methanol) in M. pulchella and water (or infusion) in H. leucopus. Data obtained from our study showed that hydrophilic extracts of M. pulchella and H. leucopus were significant sources of antioxidants and enzyme inhibition activities and that they might be used as raw materials in the food, nutraceutical, biotherapeutic and pharmaceutical industries. Graphic abstract: [Figure not available: see fulltext.].

Tomato (Solanum lycopersicum L.) is one of the most cultivated vegetable in the world and it represents a large source of bioactive compounds, including carotenoids and polyphenols (phenolic acids and flavonoids). However, the concentration of flavonoids in tomato is considered sub-optimal, particularly because anthocyanins are not generally present. Therefore, this crop has been the object of an intense metabolic engineering in order to obtain anthocyanin-enriched tomatoes by using either breeding or transgenic strategies. Some wild tomato species, such as S. chilense and S. cheesmaniae, biosynthesize anthocyanins in the fruit sub-epidermal tissue, and some alleles from those genotypes have been introgressed into a new developed purple tomato line, called “Sun Black” (SB). It is a tomato line with a purple skin color, both in green and in red fruit stages, due to the biosynthesis of anthocyanins in the peel, and a normal red color pulp, with a taste just like a traditional tomato. SB is the result of a breeding programme and it is not a genetically modified (GM) product. We report the chemical characterization and structure elucidation of the attractive anthocyanins found in the peel of SB tomato, as well as other bioactive compounds (carotenoids, polyphenols, vitamin C) of the whole fruit. Using one- and two-dimensional NMR experiments, the two main anthocyanins were identified to be petunidin 3-O-[6″-O-(4‴-O-E-p-coumaroyl-α-rhamnopyranosyl) -β-glucopyranoside]-5-O-β-glucopyranoside (petanin) and malvidin 3-O-[6″-O-(4‴-O-E-p-coumaroyl-α-rhamnopyranosyl)-β-glucopyranoside]-5-O-β-glucopyranoside (negretein). The total anthocyanins in the whole ripe fruit was 1.2 mg/g dry weight (DW); 7.1 mg/100 g fresh weight (FW). Chlorogenic acid (the most abundant phenolic acid) was 0.6 mg/g DW; 3.7 mg/100 g FW. The main flavonol, rutin was 0.8 mg/g DW; 5 mg/100 g FW. The total carotenoid content was 211.3 μg/g DW; 1,268 μg/100 g FW. The total phenolic content was 8.6 mg/g DW; 52.2 mg/100 g FW. The vitamin C content was 37.3 mg/100 g FW. The antioxidant activities as measured by the TEAC and ORAC assays were 31.6 and 140.3 μmol TE/g DW, respectively (193 and 855.8 μmol TE/100 g FW, respectively). The results show the unique features of this new tomato genotype with nutraceutical properties. publishedVersion

Fruits are an important part of the human diet and a major source of bioactive compounds (BCs) with human health benefits. Among different BCs present in fruits are polyphenols, carotenoids and some vitamins (C), which are all correlated with antioxidant potential of fruits. In the last thirty years, the genetic improvement of fruit trees has mainly been driven by productivity issues and consumers preference; the quality traits have only recently been recognized as important as the commercial ones. The nutraceutical characterization has become one issue to address when dealing with genetic improvement of fruits. The content of BCs in fruits can vary depending on different factors, primarily due to the genetic base, and the environmental conditions where fruit trees are cultivated. Effects of light, temperature, cultural practices, nutrient management, elicitors and biostimulants are presented here as outcomes on how research can contribute to enhance and improving sustainable crop production.

The olive tree &ldquo, Cellina di Nard&ograve, &rdquo, (CdN) is one of the most widespread cultivars in Southern Italy, mainly grown in the Provinces of Lecce, Taranto, and Brindisi over a total of about 60,000 hectares. Although this cultivar is mainly used for oil production, the drupes are also suitable and potentially marketable as table olives. When used for this purpose, olives are harvested after complete maturation, which gives to them a naturally black color due to anthocyanin accumulation. This survey reports for the first time on the total phenolic content (TPC), anthocyanin characterization, and antioxidant activity of CdN olive fruits during ripening and after fermentation. The antioxidant activity (AA) was determined using three different methods. Data showed that TPC increased during maturation, reaching values two times higher in completely ripened olives. Anthocyanins were found only in mature olives and the concentrations reached up to 5.3 g/kg dry weight. AA was determined for the four ripening stages, and was particularly high in the totally black olive fruit, in accordance with TPC and anthocyanin amounts. Moreover, the CdN olives showed a higher TPC and a greater AA compared to other black table olives produced by cultivars commonly grown for this purpose. These data demonstrate the great potential of black table CdN olives, a product that combines exceptional organoleptic properties with a remarkable antioxidant capacity.

Prunus mahaleb L. is a marginal fruit crop producing cherry-like dark purple drupes with a very bitter taste, rich in polyphenolic compounds. A mahaleb fruit concentrated extract ( mfce ) has been assayed for its biological activities. In this work, we report results on the in vitro effects of mfce including: i) anti-proliferative, gap junction intercellular communication (GJIC) modulation and pro-apoptotic properties in a breast cancer cell line (MCF-7), ii) anti-inflammatory properties in human vascular endothelial cells (HUVEC), iii) anti-mutagenic effect on yeast Saccharomyces cerevisiae strain D7. Mfce exerted a dose-dependent anti-proliferative effect on MCF-7 cells together with a significant time-dependent increase in GJIC and a pro-apoptotic effect. Furthermore, mfce significantly reduced levels of endothelial inflammatory antigens in a concentration-dependent manner. Finally, mfce protected S. cerevisiae cells against H 2 O 2 cytotoxicity by direct radical scavenging activity. This study demonstrated multiple healthful biological effects of mfce , highlighting it as a potential nutraceutical product.

Background The production and consumption of cherries has increased recently due to consumer awareness of their health benefits, since they are rich in polyphenolics (namely anthocyanins and hydroxycinnamic acids). Global sweet cherry production increased over the last 16 years from 1.9 to 2.32 million tons, with Turkey, USA, Iran, as the main producers. Sour cherry production has been static during the same period at around 1.1–1.3 million tons concentrated in Europe, with the Russian Federation as the predominant producer. Scope and approach This review presents a comprehensive survey of the current literature on sweet and sour cherries, from the origin and distribution of the fruit crop to their antioxidant and health properties. Moreover, possible further investigations are highlighted after summarizing the existing body of knowledge. Key findings and conclusions Sweet cherries are mostly consumed fresh, and are also highly perishable, whereas sour cherries are most frequently processed. Sweet cherries are appreciated for their organoleptic aspects, such as taste, texture and color, whereas processed sour cherries are recognized for their higher bioactive content. Both sweet and sour cherries exert beneficial health effects to counteract oxidative stress, reduce inflammation, modulate blood glucose, and enhance cognitive function. Sour cherries supplementation in particular, has been reported to benefit in recovery from exercise-induced muscle damage. The health benefits derived from consuming cherries should increase this fruit tree crop cultivation and improve the agricultural practices and food processing technology to provide consistent and reliable supply of these fruits.

In the last years, the interest in Italian monovarietal oils has increased due to their specific organoleptic qualities. Extra virgin olive oils (EVOOs) are rich in phenolic compounds, secondary metabolites well known and studied for their nutraceutical properties. However, among EVOOs, there is great variability in phenolic composition due to the origin, the production technique, and mainly, the genotype. The aim of this work was to evaluate the different phenolic profiles and the antioxidant activities of monovarietal oils. The results confirm this variability. In fact, the overall content of oleuropein varies up to four times between the different genotypes (from 33.80 to 152.32 mg/kg oil), while the oleocanthal content is significant only in two oils. The antioxidant activity, determined with 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and oxygen radical absorbance capacity (ORAC) assays, is correlated with the content of total phenolic substances, with half maximal inhibitory concentration (IC50) values for the DPPH test ranging from 160 to 91 mg of oil, while the ORAC test shows values between 5.45 and 8.03 &mu, mol Trolox equivalent (TE)/g oil.

The aim of this study was to evaluate the effect of a commercially available herbal preparation(Oxyphenol®, I-Green, Padua, Italy) ofPinus taeda hydrolyzed lignin (PTHL) on beef fattening performance, in vitro rumen apparent digestibility, volatile fatty acid (VFA) and gas production. Forty Limousine steers were randomly divided into two groups of twenty individuals each and reared in two separated pens. The control (CON) group did not receive PTHL whereas the experimental group was supplemented with PTHL (35 g/day for each for the first 90 days of the trial and 70 g/day for each for other 30 days). The data set of in vitro rumen apparent digestibility, VFA and gas production was subjected to analysis of variance (ANOVA) using the GLMprocedure in SAS software (SAS 9.4), considering the dietary treatment and the time as fixed effects. The inclusion of PTHL did not affect slaughter performance, with no differences in live weight at slaughter, daily weight gain and carcass dressing percentage between groups. In addition, the apparent digestibility of nutrients tested (dry matter, organic matter, crude fiber, NDF, crude protein and ether extract) was unaffected both by the dietary treatment and by sampling time. Rumen fluid pH was unaffected both by either dietary treatment or the sampling time,showing mean values ranging from 6.53 to 6.61. The acetate concentration was significantly greater for rumen fluid from PTHL group than in CON one, from 30 to 120 days (P< 0.01).Propionate production showed higher levels in rumen fluid from the PTHL group at 60, 90 and120 days of supplement administration (P< 0.001). Conversely, butyrate concentrations showed an opposite trend, with lower content in rumen fluid from the PTHL group, compared with CON group at 30 days (P= 0.0188). Total VFA concentration did not show significant differences because of sampling time (P> 0.05), but the dietary effect induced higher values inPTHL group from 30 to 120 days (P< 0.05). Finally, rumen fluid from PTHL during fermentation produced less CH4relative to CON, with lower values from 30 to 120 days compared to day 0 (P< 0.001). Dietary inclusion of PTHL reduced CH4production but did not affect apparent digestibility and live and slaughter performance.

Background The identification of novel plant-based functional foods or nutraceutical ingredients that possess bioactive properties with antioxidant function has recently become important to the food, nutraceutical and cosmetic industries. This study evaluates the polyphenolic composition, identifies bioactive compounds and assays the total antioxidant capacity of Prunus mahaleb L. fruits collected from different populations and sampling years in the countryside around Bari (Apulia Region, Italy). Results We identified nine polyphenolic compounds including major anthocyanins, coumaric acid derivatives and flavonols from P. mahaleb fruits. The anthocyanin content (in some populations > 5 g kg(-1) fresh weight; FW) in the fruit was comparable to that reported for so-called superfruits such as bilberries, chokeberries and blackcurrants. Coumaric acid derivatives comprised a large portion of the total polyphenolic content in the P. mahaleb fruits. Antioxidant activities, assessed using ORAC and TEAC assays, measured up to 150 and 45 mmol Trolox equivalents kg(-1) FW, respectively. Therefore antioxidant capacity of P. mahaleb fruits is relatively high and comparable to that of superfruit varieties that are often used in commercial nutraceutical products. Conclusion Our findings suggest that mahaleb fruit (currently not consumed fresh or used in other ways) could serve as a source of bioactive compounds and therefore find interest from the functional food and nutraceutical industries, as a natural food colorant and antioxidant ingredient in the formulation of functional foods. © 2015 Society of Chemical Industry.

There is an increasing consumer and retailer demand for new natural colorants. Anthocyanins are a class of molecules already used as water-soluble natural color- ant and for their health effects. Prunus mahaleb L. is a tree native to the Mediterranean area, producing highly pig- mented small-stone fruits currently not used for fresh con- sumption due to their astringent and sour taste. In the pre- sent study, we set up a protocol to obtain a "mahaleb fruit concentrated extract" (mfce) and characterized it chemi- cally by high-performance liquid chromatography/diode array detection. Anthocyanins, flavonols, flavanols, cou- marin, sugars, ascorbic acid and organic acids were identi- fied and quantified. After characterization, the antioxidant capacity of the mfce was assayed in vitro by three different methods: Trolox equivalent antioxidant capacity, Oxygen Radical Absorbance Capacity and Folin-Ciocalteu assays. The concentrated extract produced and characterized in this study showed a high content of anthocyanins, flavonols and coumarin and a strong antioxidant capacity. Therefore, mfce may be of interest as a natural food colorant and as an ingredient in the formulation of functional foods.

Betacyanin (betanin), total phenolics, vitamin C and antioxidant capacity (by Trolox-equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays) were investigated in two differently colored cactus pear (Opuntia ficus-indica (L.) Mill.) genotypes, one with purple fruit and the other with orange fruit, from the Salento area, in Apulia (South Italy). In order to quantitate betanin in cactus pear fruit extracts (which is difficult by HPLC because of the presence of two isomers, betanin and isobetanin, and the lack of commercial standard with high purity), betanin was purified from Amaranthus retroflexus inflorescence, characterized by the presence of a single isomer. The purple cactus pear variety showed very high betanin content, with higher levels of phenolics, vitamin C, and antioxidant capacity (TEAC) than the orange variety. These findings confirm the potential for exploiting the autochthonous biodiversity of cactus pear fruits. In particular, the purple variety could be an interesting source of colored bioactive compounds which not only have coloring potential, but are also an excellent source of dietary antioxidant components which may have beneficial effects on consumers’ health.

Anthocyanins, the naturally occurring pigments responsible for most red to blue colours of flowers, fruits and vegetables, have also attracted interest because of their potential health effects. With the aim of contributing to major insights into their structure–activity relationship (SAR), we have evaluated the radical scavenging and biological activities of selected purified anthocyanin samples (PASs) from various anthocyanin-rich plant materials: two fruits (mahaleb cherry and blackcurrant) and two vegetables (black carrot and “Sun Black” tomato), differing in anthocyanin content (ranging from 4.9 to 38.5 mg/g DW) and molecular structure of the predominant anthocyanins. PASs from the abovementioned plant materials have been evaluated for their antioxidant capacity using Trolox Equivalent Antioxidant Capacity (TEAC) and Oxygen Radical Absorbance Capacity (ORAC) assays. In human endothelial cells, we analysed the anti-inflammatory activity of different PASs by measuring their effects on the expression of endothelial adhesion molecules VCAM-1 and ICAM-1. We demonstrated that all the different PASs showed biological activity. They exhibited antioxidant capacity of different magnitude, higher for samples containing non-acylated anthocyanins (typical for fruits) compared to samples containing more complex anthocyanins acylated with cinnamic acid derivatives (typical for vegetables), even though this order was slightly reversed when ORAC assay values were expressed on a molar basis. Concordantly, PASs containing non-acylated anthocyanins reduced the expression of endothelial inflammatory antigens more than samples with aromatic acylated anthocyanins, suggesting the potential beneficial effect of structurally diverse anthocyanins in cardiovascular protection.

Anthocyanins, the naturally occurring pigments responsible for most red to blue colours of flowers, fruits and vegetables, have also attracted interests because of their potential health effects. With the aim of contributing to major insights into their structure-activity relationship (SAR), we have evaluated the radical scavenging and biological activities of selected purified anthocyanin samples (PASs) from various anthocyanin-rich plant materials: two fruits (mahaleb cherry and blackcurrant) and two vegetables (black carrot and ‘Sun Black’ tomato). PASs from the above-mentioned plant material have been evaluated for their antioxidant capacity, using TEAC and ORAC assays. In human endothelial cells, we analysed the biological activity of different PASs by measuring their effects on the expression of endothelial inflammatory markers, including endothelial adhesion molecules VCAM-1 and ICAM-1. We demonstrated that all the different PASs showed biological activity. They exhibited antioxidant capacity of different magnitude, higher for samples containing non-acylated anthocyanins (typical for fruits) compared to samples containing more complex anthocyanins acylated with cinnamic acid derivatives (typical for vegetables), even though this order was slightly reversed when ORAC assay values were expressed on molar basis. Concordantly, PASs containing non-acylated anthocyanins reduced the expression of endothelial inflammatory antigens more than samples with aromatic acylated anthocyanins, suggesting the potential beneficial effect of structurally diverse anthocyanins in cardiovascular protection.

Plant cell cultures as valuable tools for the production of specific metabolites can be greatly improved by the application of elicitors including cyclodextrins (CDs) for enhancing the yields of the desired plant compounds. Here the effects of 2,6-dimethyl-β-cyclodextrins (DIMEB) on the production of carotenoids and quinones from Artemisia annua L. cell suspension cultures were investigated. The addition of 50 mM DIMEB induced an early increase of intracellular carotenoid and quinone contents, which could be observed to a higher extent for lutein (10-fold), Q9 (3-fold) and Q10 (2.5-fold). Real Time PCR analysis revealed that the expression of 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR) gene in DIMEB treated cell cultures after three days was 2.5-fold higher than in untreated samples, thus suggesting that the DIMEB induced increase of carotenoids and quinones could be due to the induction of the plastidial isoprenoid biosynthetic route. In addition, the DIMEB treatment induced an enhanced release of carotenoids and quinones into the culture medium of A. annua cell suspension cultures possibly due to the ability of CDs to form inclusion complexes with hydrophobic molecules.

Eugenia myrtifolia Sims fruits have been characterized for their anthocyanin content and quality. Following the common procedures and validated methods for separation, identification and quantification of anthocyanins, we found the presence of malvidin 3,5- O -diglucoside as a unique anthocyanin (32,59 mg/100 g fresh weight of the fruit). An in vitro shoot culture of E. myrtifolia was established in order to explore the possibility to produce pigments in vitro . The presence of the only malvidin 3,5- O -diglucoside could be a useful feature in order to manipulate biotechnologically the anthocyanin biosynthetic pathway in the in vitro material (callus and suspension cultures). Industrial relevance The research interest on the phenolic composition of non-traditional food plants is a leading trend in the looking for biofunctional compounds to be included in “designer foods”. Eugenia myrtifolia , a Myrtaceae species from the South hemisphere, produces red edible fruits, that have been characterized for their anthocyanin composition, revealing the presence of the only malvidin 3,5- O -diglucoside, at relatively low amounts. Since attempts for in vitro callus production and for anthocyanin induction from the callus of E. myrtifolia have been carried out, the presence of the specific anthocyanin compound could be a useful feature in order to manipulate biotechnologically the anthocyanin biosynthetic pathway in the in vitro material, pushing the biosynthetic pathway towards the unique final product. Since malvidin is a final product (methoxylated anthocyanin) of the anthocyanin pathway, this in vitro system could help to study the regulation of the pathway, altering the genetic (regulator genes) or physiological control of the pathway. This could allow to obtain better product yields even in another in vitro systems, possibly overcoming the key bottleneck of the secondary metabolite production by plant cell and tissue cultures.

Artemisia annua L. is a herb traditionally used for treatment of fevers. The glandular trichomes of this plant accumulate, although at low levels, artemisinin, which is highly effective against malaria. Due to the great importance of this compound, many efforts have been made to improve knowledge on artemisinin production both in plants and in cell cultures. In this study, A. annua suspension cultures were established in order to investigate the effects of methyl jasmonate (MeJA) and miconazole on artemisinin biosynthesis. Twenty-two micro molar MeJA induced a three-fold increase of artemisinin production in around 30 min; while 200 μm miconazole induced a 2.5-fold increase of artemisinin production after 24 h, but had severe effects on cell viability. The influence of these treatments on expression of biosynthetic genes was also investigated. MeJA induced up-regulation of CYP71AV1, while miconazole induced up-regulation of CPR and DBR2.

The lecture will be on subject-specific topics and will try to offer excellent possibilities for exchange of experiences and source for discussion. In my presentation the importance of the anthocyanin content and other polyphenolic compounds for premium (nutraceutical) value cherries will be focused, in order to give growers the awarness of the fruit quality in the chain production.

Plant cell cultures are valuable tools for the production of bioactive compounds. Recently, many efforts have been made in order to develop and optimize strategies for increasing the yields of the desired plant metabolites by eliciting their biosynthesis or improving the efficiency of product recovery. We established Artemisia annua L. (Asteraceae) cell suspension cultures to investigate the biosynthesis of the antimalarial artemisinin (AN) [1] and other healthful phytochemicals such as carotenoids and quinones. The use of culture supplementation with cyclic oligomers ?-cyclodextrins was successful to significantly increase intracellular and extracellular levels of AN, lutein, Q9 and Q10 [2, 3]. In order to investigate the mechanism underlying these effects, other oligosaccharides having a linear structure were explored as putative elicitors in A. annua cell suspension cultures. For this purpose, oligogalacturonides (OGA) were obtained by hydrolysis of polygalacturonic acid with pectolyase from Aspergillus japonicus. Various OGA fractions were used for culture medium supplementation at different concentrations. The results showed that the 4-5 OGA fraction induced significant increases of AN and the intermediate dihydroartemisinic acid. These results suggest that the ability of ?-CDs to elicit the production of bioactive compounds in A. annua cell cultures could be due to their chemical similarity to pectic oligosaccharides often occurring in plants after fungal infection.

The identification of novel plant-based functional foods or nutraceutical ingredients that possess bioactive properties with antioxidant function has recently become important to the food, nutraceutical and cosmetic industries. This study evaluates the polyphenolic composition, identifies bioactive compounds and assays the total antioxidant capacity of Prunus mahaleb L. fruits collected from different populations and sampling years in the countryside around Bari (Apulia Region, Italy).We identified nine polyphenolic compounds including major anthocyanins, coumaric acid derivatives and flavonols from P. mahaleb fruits. The anthocyanin content (in some populations5 g kg(-1) fresh weight; FW) in the fruit was comparable to that reported for so-called superfruits such as bilberries, chokeberries and blackcurrants. Coumaric acid derivatives comprised a large portion of the total polyphenolic content in the P. mahaleb fruits. Antioxidant activities, assessed using ORAC and TEAC assays, measured up to 150 and 45 mmol Trolox equivalents kg(-1) FW, respectively. Therefore antioxidant capacity of P. mahaleb fruits is relatively high and comparable to that of superfruit varieties that are often used in commercial nutraceutical products.Our findings suggest that mahaleb fruit (currently not consumed fresh or used in other ways) could serve as a source of bioactive compounds and therefore find interest from the functional food and nutraceutical industries, as a natural food colorant and antioxidant ingredient in the formulation of functional foods. © 2015 Society of Chemical Industry.

Glutathione S-transferases (GST) are multifunctional proteins encoded by a large gene family, divided on the basis of sequence identity into phi, tau, theta, zeta and lambda classes. The phi and tau classes are present only in plants. GSTs appear to be ubiquitous in plants and are involved in herbicide detoxification and stress response, but little is known about the precise role of GSTs in normal plant physiology and during biotic and abiotic stress response. Two cDNAs representing the two plant classes tau and phi, AtGSTF9 and AtGSTU26, were expressed in vitro and the corresponding proteins were analysed. Both GSTs were able to catalyse a glutathione conjugation to 1-chloro-2,4-dinitrobenzene (CDNB), but they were inactive as transferases towards p-nitrobenzylchloride (pNBC). AtGSTF9 showed activity towards benzyl isothiocyanate (BITC) and an activity as glutathione peroxidase with cumene hydroperoxide (CumHPO). AtGSTU26 was not active as glutathione peroxidase and towards BITC. RT-PCR analysis was used to evaluate the expression of the two genes in response to treatment with herbicides and safeners, chemicals, low and high temperature. Our results reveal that AtGSTU26 is induced by the chloroacetanilide herbicides alachlor and metolachlor and the safener benoxacor, and after exposure to low temperatures. In contrast, AtGSTF9 seems not to be influenced by the treatments employed.