Carotenoids, including beta-carotene, lycopene, and derivatives, such as retinoic acid, have been studied for their significant antiproliferative and differentiating activity on cancer cells in experimental models and in clinics. We are presenting here data on the mechanism of action of a carotenoid-enriched extract obtained from the pumpkin Cucurbita moschata, variety "long of Naples," on two malignant human cell lines, Caco-2 and SAOs, derived from a colon adenocarcinoma and an osteosarcoma, respectively. The carotenoid extract has been obtained from pumpkin pulp and seeds by supercritical CO2 extraction and employed to prepare oil-in-water nanoemulsions. The nanoemulsions, applied at a final carotenoid concentration of 200-400 mu g/ml, were not cytotoxic, but induced a delay in cell growth of about 40% in both SAOs and Caco-2 cell lines. This effect was associated with the activation of a "nonprotective" form of autophagy and, in SAOs cells, to the induction of cell differentiation via a mechanism that involved AMPK activation. Our data suggest the presence of a pool of bioactive compounds in the carotenoid-enriched extract, acting additively, or synergistically, to delay cell growth in cancer cells.

Whole wheat foods are significant source of compounds exhibiting health-promoting properties. One of the most abundant class of phytochemicals in the wheat grain is represented by phenolic acids that are typically localized in the bran and germ portions. The objective of this study was to estimate the extent of variation for phenolic acids in durum wheat (T. turgidum L. ssp. durum). In addition, this study aimed at evaluating the anti-inflammatory activity of phenolic acids contained in whole-meal flour extracts. Phenolic acids were recovered from the whole meal flours of 65 durum cultivars and subsequently identified and quantified by HPLC-DAD analysis. Then, the anti-inflammatory activity of phenolic acids extracts was evaluated on LPS-stimulated HT-29 human colon cells by measuring the levels of interleukin 8 (IL-8) and transforming growth factor ?1 (TGF- ?1). A large variation for the content of phenolic acids was observed among genotypes and, on average, it accounted for 830 ?g/g dry weight. Whole meal flour extract significantly inhibited the secretion of the pro-inflammatory IL-8 mediator at 66 µg/mL of phenolic acids. Conversely, the secretion of the anti-inflammatory mediator TGF-?1 was not modified by addition of phenolic acids to HT-29 cells. Results showed that durum cultivars have different contents of phenolic acids, suggesting that a number of elite varieties could be used for breeding purposes. Moreover, results provide further insight into the health-related benefits of durum wheat foods as depending on the anti-inflammatory activity of phenolic acids.

The modern two-phase technology of the Leopard series for olive oil industrial extraction developed by the Pieralisi Group (Pieralisi Maip S.p.A., Jesi, AN, Italy) generates large quantities of a novel by-product (olive paste - OP) constituted of the partially defatted wet drupe pulp without any traces of the kernel. OP is rich in lipophilic and hydrophilic molecules able to exert beneficial effects on human health, including triterpenic acids, phytosterols, tocopherols, carotenoids, polyphenols, minerals and fibers [1]. In the perspective of by-product valorization through a modern biorefinery approach, it is worth noting that dried OP contains more than 10% oil characterized by a well-balanced lipid profile, rich in mono and polyunsaturated fatty acids, and a very good oxidative stability, due to the high concentration of fat-soluble antioxidants. This makes OP oil particularly suitable as functional ingredient for food/feed industry, as well as for the formulation of nutraceutical, cosmeceutical and pharmaceutical products. Supercritical carbon dioxide (SC-CO2) is a green and environmentally friendly technology for the effective extraction of high-value natural molecules. It is gaining a foothold in industrial production of solvent-free vegetable oils and has been also applied to concentrate oil products of lipophilic micronutrients (e.g. vitamin E in soybean and olive oils) [2]. Nevertheless, the industrial application of SC-CO2 technology requires a careful optimization of the operative parameters to make the process efficient from both an economic and a productive point of view. In this work, a response surface methodology approach, based on the Box-Behnken Design, was used to determine the optimal parameters of pressure, temperature and time to simultaneously maximize oil production from the freeze-dried OP of cultivar Leccino and concentrate the most abundant lipophilic bioactives.The experimental data were fitted to a second-order polynomial equation by multiple regression analysis, and examined using appropriate statistical methods. The 3-D response surface plots derived from the mathematical models were applied to determine the optimal extraction parameters, which resulted: temperature 70 °C, pressure 35.5 MPa and time 62 min. Under these conditions, the oil experimental yield was 14.0±0.7%, in close agreement with the predicted value. Compared to the freeze-dried OP, SC-CO2 extracted oil was much concentrated in phytosterols (~13 fold), tocopherols (~6 fold) and squalene (~8 fold); total carotenoid concentration remained almost unchanged, while triterpenic acids, being polar, were substantially reduced (- 79.5%). However, their total content was much higher than in extra virgin olive oil. The biological effects produced by OP oil intake were examined on 3 groups of healthy BALB/c mice (n=5/group) orally administered (for 4 weeks) with 20 or 40 mg/die of OP oil. Untreated animals were used as control. OP oil supplementation had no sig

Plant cell cultures are valuable bioreactors for the production of bioactive compounds. Artemisia annua L. cell suspension cultures established for producing the antimalarial artemisinin (AN) were exploited to investigate the biosynthesis of other healthful phytochemicals. Culture supplementation with the putative elicitors ß-cyclodextrins (ß-CDs) was successful to increase intracellular and extracellular levels of AN. Other bioactive isoprenoid compounds, known for their antioxidant activity, were also investigated in the supplemented A. annua cell cultures by using metabolomic and transcriptomic analyses. In particular, the application of ß-CDs was useful to induce a considerable significant increase of lutein, quinones (Q9 and Q10) intracellular contents compared to untreated cell cultures. Moreover, the release of carotenoids and quinones into the culture medium of ß-CDs-treated cell cultures was significantly enhanced. Real Time PCR analysis revealed an up-regulation of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) gene, suggesting that the ß-CDs induced increase of isoprenoid compounds could be due to the enhancement of the plastidial isoprenoid biosynthetic route. In addition, protoplasts derived from Artemisia suspension cultures were explored to engineer subcellular compartmentalization using a modified SNARE protein as a molecular tool for biotechnological metabolite accumulation.

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 potential of sweet potato as an alternative crop for bioethanol production has been assessed. We evaluated the amount of soluble sugars, starch and cell wall polysaccharides in tubers of three sweet potato cultivars characterized by different pulp and peel colouration: "Yellow yam" with yellow flesh and brown peel, "White yam" with white flesh and white peel and "Orange yam" with orange flesh and brown peel. The results confirm the high concentration of carbohydrates in sweet potato tubers, especially "Yellow yam", mainly in the form of starch (67%) and soluble sugars (26%). "Yellow yam", which is the most widespread cultivar in Salento, appeared the best choice as biomass for bioethanol production. It is characterized by high productivity (20-40tons/ha year). Results also suggest that "Yellow yam" cultivar has great potential as a bioethanol source in southern Italy with an estimated agroindustrial production yield higher than 2032l/ha year.

Artemisinin is a sesquiterpene antimalarialcompound produced, though at low levels (0.1-1% dryweight), in Artemisia annua in which it accumulates in theglandular trichomes of the plant. Due to its antimalarialproperties and short supply, efforts are being made toimprove our understanding of artemisinin biosynthesis andits production. Native ?-cyclodextrins, as well as thechemically modified heptakis(2,6-di-O-methyl)-?-cyclodextrin(DIMEB) and 2-hydroxypropyl-?-cyclodextrins,were added to the culture medium of A. annua suspensioncultures, and their effects on artemisinin production wereanalysed. The effects of a joint cyclodextrin and methyljasmonate treatment were also investigated. Fifty millimolarDIMEB, as well as a combination of 50 mMDIMEB and 100 ?M methyl jasmonate, was highlyeffective in increasing the artemisinin levels in the culturemedium. The observed artemisinin level (27 ?mol g-1 dryweight) was about 300-fold higher than that observed inuntreated suspensions. The influence of ?-cyclodextrinsand methyl jasmonate on the expression

The increased consumption of whole wheat grains and whole wheat products has been associated with reduced risk of developing chronic diseases, such as cardiovascular disease, type 2 diabetes and colon cancer. These beneficial effects have been ascribed to the presence in whole wheat kernels of bioactive compounds which may vary for total content and composition among different wheat species and wheat varieties. In this work we present the profile of hydrophilic and lipophilic bioactive compounds of whole wheat semolina from five durum wheat elite cultivars. Whole semolina samples were analyzed to evaluate the total content and composition of phenolic acids (hydrophilic extract) and the total content and composition of carotenoids, tocopherols and tocotrienols (lipophilic extract). The total phenolic acid content was variable among the cultivars and ranged from 488 ?g/g to 1490 ?g/g whole flour. Among the detected compounds, ferulic acid was the most abundant, followed by sinapic acid and p-coumaric acid. Total carotenoid content varied from 2.64 µg/g whole flour and 4.75 µg/g whole flour and were mostly represented by lutein and zeaxanthin, while ?- e ?-carotene were present in trace amounts. Three different homologues of tocotrienols were detected (?, ? and ?), varying in a range between 18.3 and 28.6 µg/g whole flour, while tocopherols were detected in trace amounts. Duilio and Svevo cultivars, exhibited the highest content of hydrophilic and lipophilic bioactive compounds, respectively, and were selected to test the anti-inflammatory activity of extracts on human intestinal HT-29 cells. Preliminary experiments were carried out in order to assess the highest not cytotoxic concentration of lipophilic and hydrophilic extracts by using MTT test. Both extracts will be used to assess the anti-inflammatory activity on HT-29 cells stimulated by LPS mitogen, by quantification of IL-8. This research shows that whole wheat semolina flours of these five cultivars varied significantly in their contents of bioactive compounds and differences in their anti-inflammatory potential might suggest the possibility that durum wheat varieties could be selected based on potential health benefits.

Tomato (Solanum lycopersicum L.) is one of the most cultivated vegetable in the worldand it is a large source of bioactive compounds, including lycopene and othercarotenoids, flavanols (quercetin) and flavanones (naringenin). However, theconcentration of flavonoids in tomato is considered sub-optimal, particularly becauseanthocyanins are not generally present. Therefore this species has been the object of anintense metabolic engineering in order to obtain anthocyanin-enriched tomatoes byusing either breeding or transgenic strategies.Some tomato-related wild species, such as S. chilense, S. cheesmaniae, S.lycopersicoides and S. habrochaites, biosynthesize anthocyanins in the sub-epidermaltissue, and some allneles from those genotypes have been introgressed into cultivatedgenetics backgrounds, resultig in a 'purple' skin color. The breeding activity pursued atTuscia University (Viterbo, Italy), aimed at combining different alleles leading toanthocyanin production, have been lead to "Sun BlackTM , a trade mark-protectedtomato line with deep purple pigmentation, due to the biosynthesis of anthocyanins inthe peel. "Sun Black" is therefore a breeding product, not a GMO product. This aspect isconsidered an added-value for the market. In fact "Sun Black" tomato has been availablein July 2015 on the Italian market, thanks to an exclusive agreement between the ownerof the trademark and Unicoop Firenze.We report here the chemical characterization and structure elucidation of the uniqueanthocyanins found in the epiderm of "Sun Black" tomato, as well as other bioactivecompounds (carotenoids, polyphenols, vitamin C) of the whole fruit.The results show the unique features of this tomato genotipe as a 'novel food' withfunctional properties.Keyword: tomato, anthocyanins, functional food

Pasta represents one of the most well-known products all over the world. It is a traditional food of Mediterrean diet and it is generally manufactured with durum wheat, that confers the excellent rheologicalproperties of dough, the superior color, the appearance and the cooking quality compared to other flours. The World Health Organization (WHO) and Food and Drug Administration (FDA) consider spaghetti pastaa good vehicle for the addition of nutrients and consumers are increasingly interested in foods containing health promoting ingredients. In this contest, the objective of the present study was to characterize thebioactive components, such as carotenoids, triterpenic acids and polyphenols, present in durum wheat semolina spaghetti enriched with an olive oil patè coming from olive oil processing chain, and dried inorder to obtain a flour (OPF) suitable for the pasta process. In particular, OPF was added to the pasta dough at 10% and 15% (w/w). Finally, spaghetti with 10% OPF, considered acceptable to the sensory panel test, were characterized for their bioactive components. Further, their bioaccessibility, after simulated gastro-intestinal digestion, was also assessed. Results showed that 10% OPF addition enriched the spaghetti in ?-tocopherol, ?- and ?-carotene, maslinic and eonolic acids. The ratio of polyunsaturated (PUFA) to saturated fatty acids (SFA) resulted higher (1.16) than spaghetti used as control (0.69). The total polyphenols content (free, conjugated and bound) in dry spaghetti samples increased of about 3 times respect to the control. In particular, the amount of total free phenolics in the enriched sample, was almost 50 times higher respect to the control spaghetti, with tyrosol and oleuropein as the most abundant. In addition, it is interesting to underline that the enriched spaghetti showed an high amount of flavonoids, such as apigenin, luteolin and quercetin already present in OPF. The phenolic acids were mainly present in the conjugated and bound fractions with the highest amount in the spaghetti with 10% OPF. Finally, the bioactive components showed a good level of biaccessibility, although during the cooking process, the 50% was naturally lost.

The olive oil is an unfavorable substrate for microbial survival and growth. Only few microorganisms use olive oil fattyacids as carbon and energy sources, and survive in the presence of olive oil anti-microbial components. In this study,we have evaluated the occurrence of microorganisms in 1-year-stored extra-virgin olive oil samples. We detected thepresence of bacterial and yeast species with a recurrence of the bacterium Stenotrophomonas rhizophila and yeastSporobolomyces roseus. We then assayed the ability of all isolates to grow in a mineral medium supplemented with acommercial extra-virgin olive oil as a sole carbon and energy source, and analyzed the utilization of olive oil fatty acidsduring their growth. We finally focused on two bacterial isolates belonging to the species Pantoea septica. Both theseisolates produce carotenoids, and one of them synthesizes bioemulsifiers enabling the bacteria to better survive/growth in this unfavorable substrate. Analyses point to a mixture of glycolipids with glucose, galactose and xylose ascarbohydrate moieties whereas the lipid domain was constituted by C6-C10 ?-hydroxy carboxylic acids.

The molecular mechanisms altered by the traditional mutation and screening approach during the improvement of antibiotic-producing microorganisms are still poorly understood although this information is essential to design rational strategies for industrial strain improvement. In this study, we applied comparative genomics to identify all genetic changes occurring during the development of an erythromycin overproducer obtained using the traditional mutate-and- screen method.Results: Compared with the parental Saccharopolyspora erythraea NRRL 2338, the genome of the overproducing strain presents 117 deletion, 78 insertion and 12 transposition sites, with 71 insertion/deletion sites mapping within coding sequences (CDSs) and generating frame-shift mutations. Single nucleotide variations are present in 144 CDSs. Overall, the genomic variations affect 227 proteins of the overproducing strain and a considerable number of mutations alter genes of key enzymes in the central carbon and nitrogen metabolism and in the biosynthesis of secondary metabolites, resulting in the redirection of common precursors toward erythromycin biosynthesis. Interestingly, several mutations inactivate genes coding for proteins that play fundamental roles in basic transcription and translation machineries including the transcription anti-termination factor NusB and the transcription elongation factor Efp. These mutations, along with those affecting genes coding for pleiotropic or pathway-specific regulators, affect global expression profile as demonstrated by a comparative analysis of the parental and overproducer expression profiles. Genomic data, finally, suggest that the mutate-and-screen process might have been accelerated by mutations in DNA repair genes. Conclusions: This study helps to clarify the mechanisms underlying antibiotic overproduction providing valuable information about new possible molecular targets for rationale strain improvement.

Here we describe the encapsulation in ?-cyclodextrins (?-CDs) of wheat bran, pumpkin and tomato oleoresins, extracted by supercritical carbon dioxide, to obtain freeze-dried powders useful as ready-to-mix ingredients for novel functional food formulation. The stability of tocochromanols, carotenoids and fatty acids in the oleoresin/?-CD complexes, compared to the corresponding free oleoresins, was also monitored over time in different combinations of storage conditions. Regardless of light, storage at 25 °C of free oleoresins determined a rapid decrease in carotenoids, tocochromanols and PUFAs. ?-CD encapsulation improved the stability of most bioactive compounds. Storage at 4 °C synergized with encapsulation in preventing degradation of bioactives. Unlike all other antioxidants, lycopene in tomato oleoresin/?-CD complex resulted to be more susceptible to oxidation than in free oleoresin, likely due to its selective sequestration from the interaction with other lipophilic molecules of the oleoresin

This review reports the use of wheat milling by-products for the extraction of high quality oil and vitamin E including our results on the exploitation of durum wheat bran as a valuable source of important healthful compounds. Wheat oil can be used as an ingredient in food, pharmaceutical or cosmetic preparations because it contains important bioactive compounds such as vitamin E, carotenoids and unsaturated fatty acids. Different methods are used for oil recovery from plant materials, such as solvent extraction, mechanical pressing or the ecofriendly supercritical carbon dioxide (SC-CO2) extraction technology. By using SC-CO2, we obtained an oil from durum wheat (Triticum durum Desf.) bran and optimized the extraction conditions to increase oil and vitamin E yields. Wheat bran, which is composed of pericarp, aleurone layer and germ, is discarded during the early stages of durum wheat milling processes to obtain a final product (semolina) that is stable over time. Maximum oil and vitamin E yields were obtained when a durum wheat bran matrix with particle size of *30 mesh and a moisture content of 2.6 % was used. The optimal conditions for oil extraction were: 300-350 bar, 60-70 °C, and 4 l min-1 gaseous CO2 flow rate for 1 h. The chemical composition (vitamin E forms, carotenoids, quinones, lipids and fatty acids) of the SC-CO2 extracted oil was analyzed and compared to that of the oil extracted by Soxhlet using hexane as solvent. The findings here reported highlight the importance of durum wheat bran as a rich source of valuable natural nutrients.

In this work a process for obtaining high vitamin E and carotenoid yields by supercritical carbon dioxide(SC-CO2) extraction from pumpkin (Cucurbita moschata Duch.) is described. The results show that the useof a vacuum oven-dried [residual moisture (8%)] and milled (70 mesh sieve) pumpkin flesh matrixincreased SC-CO2 extraction yields of total vitamin E and carotenoids of 12.0- and 8.5-fold,respectively, with respect to the use of a freeze-dried and milled flesh matrix. The addition of milled(35 mesh) pumpkin seeds as co-matrix (1:1, w/w) allowed a further 1.6-fold increase in carotenoidyield, besides to a valuable enrichment of the extracted oil in vitamin E (274 mg/100 g oil) andpolyunsaturated fatty acids. These findings encourage further studies in order to scale up the processfor possible industrial production of high quality bioactive ingredients from pumpkin useful in functionalfood or cosmeceutical formulation.

Artemisinin is an effective antimalarial compound obtained by thè aerial parts of thè herbalplant Artemisia annua L. Unfortunately, thè production of artemisinin in plants is very low (0.1-1% on a dry weight basis) and its chemical synthesis is very difficult. In recent yearsbiotechnological attempts to improve artemisinin production either in plant or geneticallyengineered microrganisms were made. A. annua celi and tissue cultures were also exploredfor thè production of artemisinin, although thè yieids obtained have been so far quite low. Wehave recently established A. annua celi cultures able to biosynthesize artemisinin and torespond to thè elicitation by methyl jasmonate (MeJA). Interestingly, amounts of artemisininproduced by these cultures were also found in thè spent culture medium. Cyclodextrins (CDs)are known to increase thè water solubility of various compounds, by forming host-guestinclusion complexes. Chemically modified CDs, such as 2,6 dimethyl-p-cyclodextrins (DIMEB)have been successfully used to improve thè production of health-promoting plant metabolitessuch as resveratrol in grape celi cultures. The aim of this work was to evaluate thè ability ofDIMEB to enhance artemisinin production in A. annua suspension celi cultures. HPLCanalysis of artemisinin and Real Time PCR gene expression analysis were carried out inDIMEB treated and untreated cultures. DIMEB induced a 300-fold increase of artemisininlevels in thè spent culture medium after a three-day-treatment. The addition of MeJA induceda further increase of thè artemisinin amounts. The expression levels of artemisininbiosynthetic genes (cytochrome P450 monoxygenase, cytochrome P450 reductase andartemisinic aldehyde A11 reductase) were not altered by thè DIMEB treatment. These resultssuggest that DIMEB improved artemisinin yieids of A. annua celi cultures, probably notaffecting thè biosynthetic pathway but enhancing thè solubility and stability of artemisininmolecules.

The aim of this study was to investigate the influence of encapsulation on the storage stability of oil extracted by supercritical carbon dioxide from a micronized durum wheat bran fraction. Wheat bran oil was encapsulated in 2% (w/v) sodium alginate beads. Encapsulated and unencapsulated oil samples were stored at 4°C or 25°C, daylight or darkness, over 90 days and, at defined time points, subjected to stability evaluation based on fatty acid hydroperoxide production, and tocopherol (?, ? and ? forms), tocotrienol (?, ? and ? forms) and carotenoid (lutein, zeaxanthin and ?-carotene) degradation. The encapsulation of the oil into alginate beads significantly increased stability, optimally when stored at 4°C, maintaining high levels of isoprenoids and low content of fatty acid hydroperoxides over 30 days of storage.

Supercritical carbon dioxide (SC-CO2) represents an effective, non-toxic technology for extracting food-grade oleoresins from a range of edible plant materials [1]. The obtained oleoresins are a rich source of bioactive molecules, mainly carotenoids, tocochromanols and polyunsaturated fatty acids (PUFAs). Most bioactives are very reactive to light, oxygen and free radical species, this greatly limits their stability and reduce their shelf-life and health benefits [2,3]. Encapsulation into micro or nano particles is the most promising technique to increase their stability over time. Cyclodextrins (CDs) are biocompatible, non-toxic cyclic oligosaccharides obtained by enzymatic conversion of starch. They can form inclusion complexes with specific guest molecules, contributing to their stabilization against the adverse effects of physicochemical agents. CDs are approved in Europe as soluble dietary fiber and novel food ingredient. In this work is reported the encapsulation in CDs of pumpkin and tomato oleoresins, extracted by SC-CO2. The stability of tocochromanols, carotenoids and fatty acids in the oleoresin/CD complexes, compared to the corresponding free oleoresins, was monitored over time in different storage conditions (25°C, dark; 25°C light; 4°C, dark). Differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) analyses, demonstrated that oleoresins/CDs have different physical characteristics from free oleoresins. Regardless of light, storage at 25°C of free oleoresins determined a rapid decrease in carotenoids, tocochromanols and PUFAs. CD encapsulation improved the stability of most bioactive compounds. Upon storage, the retention of tocochromanols, carotenoids and fatty acids was higher in oleoresin/CD complexes stored at 4°C than in the corresponding free oleoresins, indicating that encapsulation and low temperature have a synergistic effect in preventing the degradation against oxidation of bioactives. Unlike all other analyzed antioxidants, lycopene in tomato oleoresin/CD complex resulted to be more susceptible to oxidation than in free oleoresin, likely due to its selective sequestration from the interaction with other lipophilic molecules of the oleoresin. The results suggest that oleoresin/complexes, being generally more stable than free oleoresins, can be used as ready-to-mix high-quality ingredients in the preparation of innovative functional foods simultaneously enriched with carotenoids tocochromanols, PUFAs and fibers.

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-?-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.

Tableoliveshavebeena component of theMediterranean diet for centuries,with the trendfor their consumptioncurrently increasing worldwide. They are rich in bioactive molecules with nutritional, antioxidant, anti-inflammatory orhormone-like properties. In the present study, the concentrations of phenolics, triterpenic acids, carotenoids and vitamins, aswell as fatty acid profiles and antioxidant activity,were analyzed in the edible portion of black table olives (Olea europea L.) fromItalian (Cellina di Nardò and Leccino) and Greek (Kalamàta and Conservolea) cultivars fermented with selected autochthonousstarters and in the corresponding monovarietal olive oils.RESULTS: On a fresh weight basis, Cellina di Nardò and Leccino table olives showed the highest total phenolic content. Nosignificant differences were found with respect to the levels of total triterpenic (maslinic and oleanolic) acids and vitaminE among cultivars. All table olives were characterized by high amounts of oleic, linoleic and palmitic acids. Oils were richerin lipophilic antioxidants (carotenoids and tocochromanols) than table olives, which, instead, showed a higher content ofpolyphenols and triterpenic acids than oils.CONCLUSION: The present study demonstrates that fermented table olives are an excellent natural source of unsaturated fattyacids, as well as being nutritionally important health-promoting bioactive compounds.

Table olives have been a component of the Mediterranean diet for centuries and their consumption is increasing worldwide. They contain several nutritional components such as unsaturated fatty acids, vitamin E, triterpenic acids, sterols, polyphenols and carotenoids [1]. The content of these bioactive molecules depends on the olive variety, the ripening stage of the olive fruit, the cultivation conditions and the processing method [2]. There are numerous commercial preparations of table olives and in a recent work we have developed and validated a new procedure for table olive production based on the use of selected autochthonous microbial starters and a productive process optimized to mimic the microbial evolution observed during spontaneous fermentations. [3]. In this work, we characterized some bioactive compounds and coloring pigments of fermented black table olives, belonging to two Italian (Cellina di Nardò and Leccino) and two Greek (Kalamàta and Conservolea) cultivars. Greek table olives resulted richer in carotenoids, vitamin E and triterpenic acids (maslinic and oleanoic acids) than Italian cv, contents ranged from 2.28 to 6.91 µg/g fw (edible portion), 53.64 to 59.15 µg/g fw and 2.19 to 3.28 mg/g respectively. Polyphenol compounds are known to be responsible for the bitter taste of the olive fruit and most of them are involved in table olives colour changes. In all tested black table olive cultivars, polyphenols content ranged from 0.87 mg/g fw (in Conservolea) to 23.49 mg/g fw (Cellina di Nardò). Moreover, we analysed the content of tyrosol, hydroxytyrosol, oleuropein aglycone, verbascoside, isoverbascoside, rutin, pyrocathecol, luteolin glucoside, quercetin, cyanidine rutinoside and cyniadine glucoside. In particular, Cellina di Nardò is characterized by the highest levels of anthocyanin pigments such as cyanidin 3-O-glucoside (14.77 mg/g fw) and cyanidin-3-O-rutinoside (3.17 mg/g fw) responsible for the characteristic black color. The results indicated that fermented black table olives are an excellent natural source of bioactive compounds and can represent a potential important functional food.

On a global scale, jellyfish populations in coastal marine ecosystems exhibit increasing trends of abundance. High-density outbreaks may directly or indirectly affect human economical and recreational activities, as well as public health. As the interest in biology of marine jellyfish grows, a number of jellyfish metabolites with healthy potential, such as anticancer or antioxidant activities, is increasingly reported. In this study, the Mediterranean "fried egg jellyfish" Cotylorhiza tuberculata (Macri, 1778) has been targeted in the search forputative valuable bioactive compounds. A medusa extract was obtained, fractionated, characterized by HPLC, GC-MS and SDS-PAGE and assayed for its biological activity on breast cancer cells (MCF-7) and human epidermal keratinocytes (HEKa). The composition of the jellyfish extract included photosynthetic pigments, valuable ?-3 and ?-6 fatty acids, and polypeptides derived either from jellyfish tissues and their algal symbionts. Extract fractions showed antioxidant activity and the ability to affect cell viability and intercellular communication mediated by gap junctions (GJIC) differentially in MCF-7and HEKa cells. A significantly higher cytotoxicity and GJIC enhancement in MCF-7 compared to HEKa cells was recorded. A putative action mechanism for the anticancer bioactivity through the modulation of GJIC has been hypothesized and its nutraceutical and pharmaceutical potential was discussed.

The recent finding of nutrigenomics and the growing trend towards personalized nutrition encourage research directed towards innovative functional food development [1]. Dry pasta, coming widely and frequently in the Mediterranean diet, is considered an optimal, economic and easy-to-use vehicle for phytochemicals acting as nutrition enhancers or providing specific physiological functions. Pasta, made from refined wheat, lacks many of the nutrients found in whole grains. As such, it has been the object of many supplementation strategies to improve pasta nutritional value and functional attributes.Pumpkin (Cucurbita spp.) fruits and seeds have received considerable attention in recent years because of the nutritional and health benefits of the presence on many lipophylic biologically active compounds, mainly carotenoids, tocochromanols, phytosterols and PUFA [2].Supercritical carbon dioxide (SC-CO2) pumpkin extracted oils contain the broad lipophilic phytocomplex of the starting matrix in a highly bioavailable form [3]. Phytocomplex constituents, alone or synergistically, act as antioxidants, vitamins, anti-inflammatory and anticancer agents, as well as in cell signaling and gene expression regulation, playing a pivotal role in building and maintaining our health [4]. The limited stability of the oils over time and/or during food processing and storage is the main drawback for their usage in the formulation of functional products. Most bioactives are, in fact, very reactive to light, oxygen and free radicals and/or susceptible to thermal degradation. Encapsulation into micro or nano particles is the most promising technique to increase oil stability. The feasibility of chlatrating the oil in ?-Cyclodextrin (?-CD) complexes to improve its chemical stability and prepare powders useful as ready-to-mix ingredients for novel functional food formulations was also demonstrated [5].The aim of this study was to explore the feasibility of producing an innovative supplemented durum wheat pasta enriched in the lipophilic phytocomplex extracted by SC-CO2 from ripe pumpkin peponides. The oil was mixed to the dough as free or as ready-to-mix oil/?-CDs powder. Four types of pasta were prepared: (i) control spaghetti (S-CTRL); (ii) spaghetti supplemented with ?-CDs (S-?-CD); (iii) spaghetti supplemented with pumpkin oil (S-Oil) and (iv) spaghetti supplemented with the pumpkin oil/?-CD powder (S-Oil/?-CD). The biochemical, antioxidant, textural and sensory attributes of pasta types were evaluated and compared. The results showed that S-Oil and S-Oil/?-CD spaghetti were significantly enriched in phytosterols, squalene, carotenoids, tocochromanols and unsaturated fatty acids, providing a substantial proportion of the recommended daily allowance (RDA) for A and E vitamins, as well as increased lipophilic antioxidant capacity. Furthermore, ?-CD pumpkin oil complexation increased the stability of some of the bioactive components. With regard to the sensory evaluation o

Wheat species contain a variety of polyphenols including phenolic acids, flavonoids, proanthocyanidins, condensed tannins, catechins and lignans. Phenolic acids represent the most common phenolic compounds found in whole wheat grains and can be found as soluble free, soluble conjugated bound to low molecular mass components (e.g. saccharides or organic acids); and insoluble bound forms, linked to polymers of the plant cell wall. Despite their high value for human health, only few studies have been carried out on the genetics and genomics of these bioactive compounds in durum wheat. The objectives of the present work were i) the estimation of genetic variability for phenolic acids composition and concentration in a set of 111 tetraploid wheat genotypes (Triticum turgidum L.), comprising durum cultivars, landraces and wild accessions; and ii) the detection of regions attributable to individual phenolic acids and total phenolic acids concentration by the means of a genome wide association study (GWAS). Phenolic acids were extracted from whole-meal flour and identified by DAD-HPLC analysis. A total of six major phenolic acids were quantified across the durum wheat genotypes, namely: ferulic, sinapic, p-coumaric, vanillic, syringic and p-hydroxibenzoic acids. The amount of total bound phenolic acids, ranged from 341 to 1700 ?g g-1 of whole-meal flour, with a mean value of 800 ?g g-1. The soluble free fraction (measured spectrophotometrically after Folin Ciocalteu reaction) ranged from 1280 to 3150 ?g g-1 as ferulic acid equivalents. The analysis of variance (ANOVA) revealed the effects of genotype, year and year x genotype that were significant for all phenolic acids. The ratio of genotypic variance to total variance was moderately high suggesting that phenolic acids concentration can be further improved by breeding approaches in durum wheat.The GWAS was based on the detection of correlations between genotype and phenotype in the durum germplasm collection by using a high-density wheat SNP iSelect array including approximately 90,000 gene associated SNPs. Results showed several significant marker-trait associations (MTA), identifying eight quantitative trait loci (QTL) associated with phenolic acids concentration. The highest number of MTAs was identified on chromosome 7A, where one QTL region was associated with phenolic acids content. The lowest number of MTAs was detected on chromosomes 3A and 5B, where only one MTA identified a single locus. Conservation of synteny between SNPs and the annotated genes and proteins in Brachypodium distachyon, Oryza sativa and Sorghum bicolor allowed the identification of two QTLs coincident with two different candidate genes.

Whole wheat foods are significant source of compounds exhibiting health-promoting properties. One of the most abundant class of phytochemicals in the wheat grain is represented by phenolic acids that are typically localized in the bran and germ portions. The objective of this study was to estimate the extent of genetic variation for phenolic acids in durum wheat (T. turgidum L. ssp. durum). In addition, this study aimed at evaluating the anti-inflammatory activity of phenolic acids contained in whole-meal flour extracts. Phenolic acids were recovered from the whole meal flours of 65 durum cultivars and subsequently identified and quantified by HPLC-DAD analysis. Then, the anti-inflammatory activity of phenolic acids extracts was evaluated on LPS-stimulated HT-29 human colon cells by measuring the levels of interleukin 8 (IL-8) and transforming growth factor ?1 (TGF- ?1). A large variation for the content of phenolic acids was observed among genotypes and, on average, it accounted for 830 ?g/g dry weight. Whole meal flour extract significantly inhibited the secretion of the pro-inflammatory IL-8 mediator at 66 µg/mL of phenolic acids. Conversely, the secretion of the anti-inflammatory mediator TGF-?1 was not modified by addition of phenolic acids to HT-29 cells. Results showed that durum cultivars have different contents of phenolic acids, suggesting that a number of elite varieties could be used for breeding purposes. Moreover, results provide further insight into the health-related benefits of durum wheat foods as depending on the anti-inflammatory activity of phenolic acids.

Phenolic acid intake through the consumption of whole-wheat foods provides important health benefits associated with reduced risks of cardiovascular diseases and colon cancer. The genetic variation for phenolic acids was extensively studied in common wheat, but a comprehensive survey in tetraploid wheat is lacking. In this study we evaluated the genetic variability for individual and total phenolic acids concentration existing in a large collection of tetraploid wheat (Triticum turgidum L.). A two-year evaluation was undertaken on the whole-meal flour of 111 genotypes belonging to seven T. turgidum subspecies including cultivars, landraces and wild accessions. Durum cultivars [T. turgidum subsp. durum (Desf.) MacKey], had the highest average concentration of total phenolic acids (828.7 ug g-1 dm in 2012; 834.5 ug g-1 dm in 2013) with values varying from 550.9 ug g-1 dm to 1,701.2 ug g-1 dm, indicating a variation of >3 fold. The lowest concentration of phenolic acids was found in T. turgidum subsp. dicoccum (Schrank ex Schübler) Thell. Rivet wheat (T. turgidum L. subsp. turgidum) had phenolic acid concentrations similar to those in durum, but less variation was noted among the accessions. On the other hand, the accessions of the four remaining subspecies showed lower phenolic acid concentrations and variation among the accessions as compared to durum. A total of six phenolic acids were identified across the wheat genotypes. The effects of genotype, year and year x genotype were estimated by ANOVA and resulted significant for all phenolic acids. The ratio of genotypic variance to total variance suggested the possibility of improving phenolic acid content in elite wheat germplasm through appropriate breeding programs. Moreover, significant correlations between phenolic acids and other quality characteristics of the grain were detected.

Phenolic acids are major components of plant cell walls in wheat and have important implications on human health as antioxidants with anti-tumor activity. They occur in wheat grains as: i) soluble free phenolic acids; ii) soluble conjugated phenolics bound to low molecular mass components such as saccharides or organic acids; iii) insoluble bound forms of phenolics, linked to polymers of the plant cell wall (Li et al., 2008). Despite their high value for human health, few studies have been carried out on the genetics of phenolic acids in durum wheat. The genetic variability of phenolic acids composition and concentration was investigated, over two years, in a set of 111 tetraploid wheat genotypes, belonging to seven Triticum turgidum L. subspecies, including cultivars, landraces and wild accessions. Regions attributable to individual phenolic acids and total phenolic acids concentration were identified through a genome wide association study (GWAS). A total of six phenolic acids were identified by DAD-HPLC analysis across the 111 wheat genotypes, namely: ferulic, sinapic, p-coumaric, vanillic, syringic and p-hydroxibenzoic acids. The amount of total bound phenolic acids, ranged from 341 to 1700 ?g g-1 of whole-meal flour, with a mean value of 800 ?g g-1 (Laddomada et al, 2016). The soluble free fraction (measured spectrophotometrically after Folin Ciocalteu reaction) ranged from 1280 to 3150 ?g g-1 as ferulic acid equivalents. The effects of genotype, year and year x genotype were estimated by ANOVA and resulted significant for all phenolic acids. The ratio of genotypic variance to total variance was moderately high suggesting that a breeding approach could be considered to increase phenolic acids concentration in durum wheat.The GWAS revealed a total of 29 significant marker-trait associations (MTA), identifying eight quantitative trait loci (QTL) associated with phenolic acids content. The highest number of MTAs was identified on chromosome 7A, where one QTL region was associated with phenolic acids content, while the lowest number of MTAs was detected on chromosomes 3A and 5B, where only one MTA identified a single locus. Conservation of synteny between SNP marker sequences and the annotated genes and proteins in Brachypodium distachyon, Oryza sativa and Sorghum bicolor allowed the identification of two QTLs coincident with two different candidate genes.

Lycopene is a carotenoid pigment found in a wide variety of vegetables and fruits. More than six hundred carotenoids have been identified in nature, but only twenty are present in human blood and tissues, including lycopene (the most abundant in plasma), ?-carotene, lutein and zeaxanthin [1]. Several exogenous pigmented antioxidants act on cancer progression and have immune-modulating effects by free radical scavenging activity and/or still unknown mechanisms. However, the controversy recently arisen for some of them requires a careful investigation in relation to their source and synergistic effects [2]. The purpose of this work was: i) to optimize the extraction of lycopene from ripe fruits of tomato, watermelon and GAC by supercritical CO2 in order to obtain natural, solvent-free lycopene containing oleoresins suitable as ingredients for the formulation of innovative functional foods and/or nutraceuticals; ii) to characterize each oleoresin with respect to carotenoids, tocochromanols, lipids, lipophilic and hydrophilic antioxidant activities, micro and macro elements; iii) the preparation of inclusion complexes (ICs) of the oleoresins in food grade ?-cyclodextrins (?-CDs) as hydrophilic nanocarriers [3]; iv) to assess, in-vitro, the effects of administration of each oleoresin/?-CD IC at different concentrations (0.5 to 10 µM) on cell apoptosis and on Interleukin-8 (IL-8) release by A549 human lung adenocarcinoma cells, for 24, 48 and 72 hours.The results showed that all oleoresins represent a healthy and high added-value source of lycopene characterized by: i) the absence of toxic impurities such as solvents, pesticides and heavy metals; ii) a high lycopene bioavailability, increased by the presence of co-extracted natural lipophilic components; iii) a high lipophilic antioxidant activity, improved by synergy with other biomolecules. When administered at a concentration of 10 µM for 72 hours, the three lycopene extracts showed conflicting behaviors: watermelon oleoresin increased cell apoptosis and decreased the release of IL-8, a pro-inflammatory cytokine involved in cancer progression; GAC and tomato oleoresins, instead, decreased cell apoptosis and increased IL-8 release. This discrepancy is likely related to a different lycopene/vitamin E ratio, with watermelon oleoresin showing the highest value among the extracts. Shorter incubation time and lower lycopene concentrations did not affect cell viability and IL-8 release. Further studies are required to assess the possible use of the lycopene containing oleoresin/?-CD ICs in health promoting functional foods and nutraceuticals.

The yeast Brettanomyces bruxellensis, generally considered the main oenological spoilage microbe, is able to survive during the winemaking process and it confers off-odors to wine, in reason of its ability to produce considerable amounts of volatile phenols. Forty-eight isolates of B. bruxellensis, obtained from several wines collected in Apulia (Southern Italy), were genetically characterized using an integrated approach, including a strain biodiversity analysis by Sau-PCR. Furthermore, the production of volatile phenols was assessed in wine and in synthetic medium, confirming the oenological spoilage potential of the analysed strains. Our findings indicate a remarkable genetic variability of the B. bruxellensis identified strains and corroborate the evidence of a high level of genotypic and phenotypic polymorphism within B. bruxellensis species. Moreover, the observation reported suggest that strains from wines produced in the same geographical areas often clustered differently, indicating a complex intraspecific biodiversity in the regional wine environments. Diversity in volatile phenol production reflects intraspecific biodiversity highlighted by Sau-PCR. Strains diversity linked to differences in 'spoilage potential' increase the industrial relevance of this study, allowing the design of new strategies for B. bruxellensis control in wines.

The polyphenol oxidase (PPO) enzyme, which can catalyze the oxidation of phenolics to quinones, has been reported to be involved in undesirable browning in many plant foods. This phenomenon is particularly severe in artichoke headswounded during the manufacturing process. A full-length cDNA encoding for a putative polyphenol oxidase (designated as CsPPO) along with a 1432 bp sequence upstream of the starting ATG codon was characterized for the first time from [Cynara cardunculus var. scolymus (L.) Fiori]. The 1764 bp CsPPO sequence encodes a putative protein of 587 amino acids with a calculated molecular mass of 65,327 Da and an isoelectric point of 5.50. Analysis of the promoter region revealed the presence of cis-acting elements, some of which are putatively involved in the response to light and wounds. Expression analysis of the gene in wounded capitula indicated that CsPPO was significantly induced after 48 h, even though the browning process had started earlier. This suggests that the early browning event observed in artichoke heads was not directly related to de novo mRNA synthesis. Finally, we provide the complete gene sequence encoding for polyphenol oxidase and the upstream regulative region in artichoke

Artemisia annua L., is a herbal plant traditionally used for the treatment of fever. 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 the 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 treatments on artemisinin biosynthesis. Twenty-two mM MeJA induced a three-fold increase of artemisinin production soon after 30 min. Two hundred mM miconazole induced a 2.5 fold increase of artemisinin production after 24 h treatment but had severe effects on cell viability. The influence of these treatments on the expression of biosynthetic genes was also investigated. MeJA induced an up-regulation of CYP71AV1 while miconazole induced an up-regulation of CPR and DBR2.

Table olives are one of the most important traditional fermented vegetables in Europe and their world consumption is increasing. In the Greek system, table olives are produced by natural fermentation process, that is not predictable and strongly influenced by the physical-chemical conditions and by the presence of microorganisms contaminating the olives, In this study , we have developed and validated a novel procedure for table olive production based on the use of a mixed yeast/bacteria starter.

Table olives represent one important fermented product in Europe and, in the world, their demand is constantly increasing. At the present time, no systems are available to control black table olives spontaneous fermentation by the Greek method. During this study, a new protocol for the production of black table olives belonging to two Italian (Cellina di Nardò and Leccino) and two Greek (Kalamàta and Conservolea) cultivars has been developed: for each table olive cultivar, starter-driven fermentations were performed inoculating, firstly, one selected autochthonous yeast starter and, subsequently, one selected autochthonous LAB starter. All starters formulation were able to dominate fermentation process. The olive fermentation was monitored using specific chemical descriptors able to identify a first stage (30 days) mainly characterized by aldehydes; a second period (60 days) mainly characterized by higher alcohols, styrene and terpenes; a third fermentation stage represented by acetate esters, esters and acids. A significant decrease of fermentation time (from 8 to 12 months to a maximum of 3 months) and an significant improvement in organoleptic characteristics of the final product were obtained. This study, for the first time, describes the employment of selected autochthonous microbial resources optimized to mimic the microbial evolution already recorded during spontaneous fermentations.

Two approaches were developed in order to select microorganisms suitable to be used in olive millwastewaters bioremediation. By the first approach, three hundred yeasts were isolated from fiveindustrial mills and identified by molecular analysis. The different strains were selected accordingto their capacity to grow in OMW (olive mill wastewaters) as the sole carbon source and to reducephenolics, chemical oxygen demand (COD) and antimicrobial compounds. One Geotrichum candidumisolate was used to set up a whole-cell immobilization system in calcium alginate gel andthe COD and phenolic reduction obtained using the immobilized cells showed respectively a 2.2-and 2-fold increase compared to the removal obtained using free cells. By the second approach, anew protocol was developed to isolate and select aerobic microorganisms from different industrialsamples and environmental niches (soils, OMWs) and able to detoxify olive mill wastewaters.

Two approaches were developed in order to select microorganisms suitable to be used in olive mill wastewaters bioremediation. By the first approach, three hundred yeasts were isolated from five industrial mills and identified by molecular analysis. The different strains were selected according to their capacity to grow in OMW (olive mill wastewaters) as the sole carbon source and to reduce phenolics, chemical oxygen demand (COD) and antimicrobial compounds. One Geotrichum candidum isolate was used to set up a whole-cell immobilization system in calcium alginate gel and the COD and phenolic reduction obtained using the immobilized cells showed respectively a 2.2- and 2-fold increase compared to the removal obtained using free cells. By the second approach, a new protocol was developed to isolate and select aerobic microorganisms from different industrial samples and environmental niches (soils, OMWs) and able to detoxify olive mill wastewaters.

Given the significant increase in consumer interest in organic food products, there is a need to determine to what extent there is a scientific basis for claims made for organic products. Apart from the sensory quality and food safety, there is a need to assess the nutritional value of organic vs. conventional (or integrated) food. Measurement of antioxidant activity using biologically relevant assays isimportant in the screening of fruits for potential health benefits. ORAC assay has been used in the assessment of antioxidant activity of strawberry extracts, from organically and Integrated Pest Management (IPM) strawberry plants, grown in the highly suited area of Basilicata (south Italy). Results on two genotypes ('Candonga' and 'Ventana') indicated that strawberries organically cultivated had statistically significant higher antioxidant activity than the IPM strawberries. Particularly, in the methanolic extract of organic 'Candonga', at red stage of ripening, the ORAC value was 31.5% higher than the IPM one. Further investigations on bioactive compound composition (namely anthocyanins and other flavonoids) are needed to identify the specific contribution to antioxidant capacity of organic strawberries.

Lycopene is used for several industrial applications. Supercritical CO2 (SC-CO2) extraction from red-ripe tomatofruits is an excellent technique to replace the use of harmful solvents. In this study, starting from red-ripe tomatoes of ordinaryand high-lycopene cultivars, the effect of different agronomical and technical aspects on lycopene content, stability and yieldwas evaluated throughout the production process from fresh tomatoes to the final SC-CO2-extracted oleoresin containinglycopene.RESULTS: Red-ripe tomato cultivars differed in their lycopene content. Irrigation excess or deficit caused an increase in theamount of lycopene in the fruits. Fresh tomatoeswere processed into a lyophilised matrix suitable for SC-CO2 extraction,whichcould be stored for more than 6months at -20 oC without lycopene loss. Under the optimal extraction conditions, efficienciesof up to 80% were achieved, but the recovery of lycopene in the extracted oleoresin was very low (~24%). Co-extraction of thetomato matrix mixedwith a lipid co-matrix allowed the recovery of~90% of lycopene in the oleoresin. Using the high-lycopenecultivars, the yield of total extracted lycopene increased by ~60% with respect to the ordinary cultivars. Lipids and otherbiologically active molecules were present in the oleoresin.CONCLUSION: A method for extracting, from a tomato matrix, a natural and solvent-free oleoresin containing lycopenedissolved in a highly unsaturated vegetable oil has been described. The oleoresin represents an excellent product for testingon cancer and cardiovascular disease prevention.

The high content of bioactive molecules in bran and germ portions of the kernel makes whole wheat grain a functional food to prevent several degenerative and chronic diseases. In particular, whole wheat caryopsides contain high amounts of phenolic acids having favourable effects on human health due to their antioxidant and anti-inflammatory activity. Genetic variation for phenolic acid content and composition has been documented among different cereal grains with particular regard to soft, medium and hard wheat (Triticum aestivum L.). Less attention has been devoted to tetraploid wheats (Triticum turgidum L.). The objective of this study was to evaluate the variability for phenolic acid content and composition in a core collection of 112 genotypes of tetraploid wheat, including cultivars, landraces and wild accessions. In particular, the core collection comprised 65 durum cultivars (T. turgidum L. ssp. durum), 12 accessions of ssp. turgidum, 8 of ssp. turanicum, 8 of ssp. polonicum, 3 of ssp. carthlicum, 9 of ssp. dicoccum and 7 of ssp. dicoccoides.Phenolic acids extracted from the whole semolina flour were identified and quantified by HPLC-DAD analysis. The results showed a significant genotype influence on the content of phenolic acids. The ssp. durum cultivars showed a higher content of phenolic acids compared to landraces and wild accessions. On average, total phenolic acid content ranged from about 830 ?g/g dry weight (ssp. durum) to 570 ?g/g dry weight (ssp. dicoccum and ssp. turgidum). Large and significant variation for total phenolic acid content was observed among the durum wheat cultivars, with cv. Ambral, Pedroso, Primadur, Mida and Tito largely exceeding 1000 ?g/g dry weight of total phenolic acids. Ferulic acid was the most abundant phenolic acid in all genotypes, particularly in the ssp. durum (on average 654 ?g/g dry weight), whereas the ssp. dicoccum, turgidum and diccoides showed the least content (less than 450 ?g/g dry weight). Sinapic acid was the second phenolic acid for abundance and it varied from ~54 ?g/g dry weight (ssp. carthlicum) to ~140 ?g/g dry weight (ssp. durum). Other phenolic acids identified in this work were p-coumaric acid (19 ÷ 26 ?g/g dry weight), vanillic acid (5 ÷ 9 ?g/g dry weight), syringic acid (2 ÷ 7 ?g/g dry weight) and 2,4- dihydroxybenzoic acid (1 ÷ 3 ?g/g dry weight). In conclusion, the results showed that the content of phenolic acids in the studied tetraploid wheat collection varied significantly, suggesting that a number of durum wheat elite varieties could be used for breeding purposes. Also, the use of cultivars with high phenolic acid content could allow the production of whole durum wheat food products with enhanced health-related benefits.

Table olives are one of the most important traditional fermented vegetables in Europe and their world consumption is constantly increasing. In the Greek style, table olives are obtained by spontaneous fermentations, without any chemical debittering treatment. Evolution of sugars, organic acids, alcohols, mono and polyphenol compounds and volatile compounds associated with the fermentative metabolism of yeasts and bacteria throughout the natural fermentation process of the two Italian olive cultivars Cellina di Nardò and Leccino were determined. A new protocol was developed and applied aimed at the technological characterization of LAB and yeast strains as possible candidate autochthonous starters for table olive fermentation from Cellina di Nardò and Leccino cultivars. The study of the main physical, chemical and aromatic parameters during fermentation helped to determine chemical descriptors that may be suitable for monitoring olive fermentation. In both the analyzed table olive cultivars, aldehydes proved to be closely related to the first stage of fermentation (30 days), while higher alcohols (2-methyl-1-propanol; 3-methyl-1-butanol), styrene, and o-cymene were associated with the middle stage of fermentation (90 days) and acetate esters and acetic acid with the final step of olive fermentation (180 days).

Table olives are one of the most important traditional fermented vegetables in Europe and their worldconsumption is constantly increasing. Conservolea and Kalam
ata are the most important table olivesGreek varieties. In the Greek system, the final product is obtained by spontaneous fermentations,without any chemical debittering treatment. This natural fermentation process is not predictable andstrongly influenced by the physical-chemical conditions and by the presence of microorganismscontaminating the olives. Natural fermentations of Conservolea and Kalam
ata cultivars black olives were studied in order to determine microbiological, biochemical and chemical evolution during the process. Following the process conditions generally used by producers, in both cultivars, yeasts were detected throughout the fermentation, whereas lactic acid bacteria (LAB) appeared in the last staged of the process. A new optimized specific protocol was developed to select autochthonous yeast and LAB isolates that can be good candidates as starters. These microorganisms were pre-selected for their ability to adapt to model brines, to have beta-glucosidase activity, not to produce biogenic amines. Chemical compounds deriving by microbiological activities and associated to the three different phases (30, 90 and 180 days) of the fermentation process were identified and were proposed as chemical descriptors to follow the fermentation progress.

In this study, the quali-quantitative composition of hydrophilic (phenolic acids)and lipophilic (isoprenoids) extracts from whole-meal flour of five elite Italian durum wheatcultivars was determined. Significant differences in the content of bioactive compoundswere observed among the wheat extracts, in particular concerning the content of boundphenolic acids, lutein and ?-tocotrienols. The cultivars Duilio and Svevo showed the highestamount of phenolic acids and isoprenoids, respectively. Extracts were evaluated for theiranti-inflammatory activity on HT-29 human colon cells by measuring the levels ofinterleukin 8 (IL-8) and transforming growth factor ?1 (TGF-?1). Durum wheat extractssignificantly inhibited the secretion of the pro-inflammatory IL-8 mediator at 66 ?g/mLof phenolic acids and at 0.2 ?g/mL of isoprenoids. Conversely, the secretion of theanti-inflammatory mediator TGF-?1 was not modified by neither hydrophilic nor lipophilicextracts. These results provide further insight into the potential of durum wheat on humanhealth suggesting the significance of varieties with elevated contents of bioactive components.

This study provides information about the carbohydrate present in tomato pomace (skins, seeds, and vascular tissues) as well as in the byproducts of the lycopene supercritical carbon dioxide extraction (SC-CO2) such as tomato serum and exhausted matrix and reports their conversion into bioethanol. The pomace, constituting approximately 4% of the tomato fruit fresh weight, and the SC-CO2-exhausted matrix were enzyme saccharified with 0.1% Driselase leading to sugar yields of similar to 383 and similar to 301 mg/g dw, respectively. Aliquots of the hydrolysates and of the serum (80% tomato sauce fw) were fermented by Saccharomyces cerevisiae. The bioethanol produced from each waste was usually >50% of the calculated theoretical amount, with the exception of the exhausted matrix hydolysate, where a sugar concentration >52.8 g/L inhibited the fermentation process. Furthermore, no differences in the chemical solubility of cell wall polysaccharides were evidenced between the SC-CO2-lycopene extracted and unextracted matrices. The deduced glycosyl linkage composition and the calculated amount of cell wall polysaccharides remained similar in both matrices, indicating that the SC-CO2 extraction technology does not affect their structure. Therefore, tomato wastes may well be considered as potential alternatives and low-cost feedstock for bioethanol production.

Regina tomato, a locally cultivated Italian landrace, is listed as an item in the 'List ofTraditional Agri-Food Products' of the Italian Department for Agriculture and itemised as 'SlowFood presidium' by the Slow Food Foundation. It is classified as a long-storage tomato since it can bepreserved for several months after harvest thanks to its thick and coriaceous skin. Three ecotypeswere investigated for main physical and chemical traits both at harvest and after three months ofstorage. Experimental results indicate that this tomato landrace has a qualitative profile characterizedby high concentrations of tocopherols, lycopene and ascorbic acid (maximum 28.6 and 53.7 mg/kgfresh weight, FW, and 0.28 mg/g FW, respectively) even after a long storage time, together withlower average Total Soluble Solids. The initial and post-storage contents of the bioactive compoundschanged at a different rate in each ecotype (i.e., in Monopoli Regina tomato the highest content of-Tocopherol, thereafter reduced to the same level of the other two ecotypes). These results indicateunique and unmistakable features of this long-storage tomato, closely linked to the geographicorigin area that include both natural (available technical inputs) and human (specific culturalpractices) factors.

Tribulus terrestris L. (Zygophyllaceae) is an annual plant commonly known as Puncture vine. It is dramatically gaining interest as a rich source of saponins. T. terrestris is a promising ingredient for many industries and recent patents on dermatological applications support the use of this plant for cosmetics and hygiene. Nonetheless problems arise in the selection of the material to be used. The extracts of different origins may differ substantially. Natural speciation processes normally influence 'variations' in wild-crafted medicinal plants. The genus Tribulus is emblematic. Taxonomic status of T. terrestris is complicated by the wide geographical distribution leading to high levels of genetic polymorphism. Being aware of such variability we selected 3 commercial Tribulus extracts and compared their biological effect on Candida albicans with the effect produced by an extract from local plants (South of Apulia, Italy). One of the commercial extracts with the best anti-candida performance was used to substitute triclosan in a detergent formulation and it proved to improve the product performance in the control of potentially pathogenic skin flora such as C. albicans. © Di Sansebastiano et al.

Asparagus represents a nutritious and refined food being very popular to consumers. The effects on the quality of spears cooked by conventional (boiling, steaming and microwaving) and different sous vide (using hot water or microwave) methods were analyzed. Physical, chemical and sensory traits were compared. Microwaving caused the largest weight change, the highest dry weight increase and the highest Total Color Difference in cooked samples compared to raw asparagus spears. Sous vide-microwaving showed an increase in greenness (the highest value of h°), better than the raw spears, and the lowest reduction in chlorophyll contents. After sous vide microwawing violaxanthin increased by about 42%, while after steaming neoaxanthin decreased by about 57%. Following sensory analysis, all cooked samples generally resulted acceptable (scores > 5 in a 1-9 hedonic scale), nevertheless sous vide-microwaved asparagus satisfied consumer acceptability more than the other cooked samples, especially when compared to steamed samples. Results indicate the sous vide-microwaving technique as optimal to preserve several traits, including the organoleptic ones, essential for the quality of cook-chilled asparagus spears. They also provide product-specific information usually required for cooking process strategies in the industrial area of ready-to-eat vegetables.

Pomegranate, tomato and grape seeds are quantitatively relevant agri-food by-products rich in molecules beneficialto human health. To valorize this resource, the composition and antioxidant activity of seeds and derivingsupercritical CO2 (SC-CO2) extracted oleoresins were evaluated. Grape seeds showed the highest content of totalphenolic compounds (33.9 mg GAE/g), flavonoids (15.6 mg CE/g) and condensed tannins (14.0 mg CE/g), whiletomato seeds presented the highest content of tocochromanols (159.6 ?g/g). Grape seeds showed the highesttotal antioxidant activity (178.2 ?mol TE/g), as evaluated by TEAC assay, followed by pomegranate (19.8 ?molTE/g) and tomato (9.8 ?mol TE/g). Oleoresin yields obtained by SC-CO2 extraction from the seeds ranged between3.1 (pomegranate) to 7.8 (tomato) g oleoresin/100 g. Total tocochromanols were abundant in pomegranate(2008 ?g/g) and tomato (1769 ?g/g) oleoresins; a relatively low amount was instead detected in theoleoresin extracted from grape seeds (636 ?g/g). Carotenoids were not detected in all oleoresins. Pomegranateoleoresin had a higher antioxidant activity than the others. Mono- and polyunsaturated fatty acids were moreabundant than saturated in all oleoresins, with the highest percentage of unsaturated fatty acids detected inpomegranate seed oleoresin (~90%), mainly due to punicic acid (~70%).

Lycopene is a red carotenoid thought effective in preventingsome types of cancers and modulating immune and inflammatory responses.Though some fruits share the common feature of synthesizing and storinglycopene as main carotenoid within chromoplasts, they significantlydiffer in the lipophilic phytocomplex. The purpose of this work was: i)to extract solvent-free lycopene-rich oleoresins from gac, tomato andwatermelon ripe-fruits by supercritical CO2 suitable for functional foodand nutraceutical formulations; ii) characterize the oleoresins withrespect to carotenoids, tocochromanols, lipids, metals and antioxidantactivity; iii) prepare stable aqueous suspensions through oleoresinclathration into ?-cyclodextrins (?-CDs); iv) investigate the effect ofeach lycopene-containing suspension on viability of human lungadenocarcinoma cells.The results showed that all oleoresins represent a safe added-valuesource of lycopene characterized by high lipophilic antioxidant activity,improved by the synergy with other biomolecules. Interestingly, whenadministered at identical concentrations (0.5-10 ?M) of lycopene, theextracts showed conflicting behaviors: gac and tomato suspensionssignificantly increased cell viability, whereas watermelon oleoresin/?-CDsuspension significantly decreased cell viability. Although furtherstudies are required, this research suggests a fundamental role ofphytocomplex in affecting the responses of cancer cell survival to the invitro administration of lycopene.

Strain SPC-1T was isolated from the phyllosphere of Cynara cardunculus L. var. sylvestris (Lamk) Fiori (wild cardoon), a Mediterranean native plant considered the wild ancestor of the globe artichoke and cultivated cardoon. This Gram-negative, catalase-positive, oxidase-negative, non-spore-forming, rod-shaped and non-motile strain secreted copious amounts of an exopolysaccharide and formed slimy, viscous, orange-pigmented colonies and grew optimally at around pH 6.0-6.5 and 26-30°C in the presence of 0-0.5% NaCl. Phylogenetic analysis based on comparisons of 16S rRNA gene sequences demonstrated that SPC-1T clustered together with species of the genus Sphingomonas sensu strictu. The G+C content of the DNA (66.1 mol%), the presence of Q-10 as the predominant ubiquinone, sym-homospermidine as the predominant polyamine, and 2-hydroxymiristic acid (14:0 2-OH) as the major hydroxylated fatty acid, the absence of 3-hydroxy fatty acids and the presence of sphingoglycolipid supported this taxonomical position. 16S rRNA gene sequence analysis showed that SPC-1 was most closely related to Sphingomonas hankookensis ODN7T, Sphingomonas insulae DS-28T and Sphingomonas panni C52T (98.19%, 97.91% and 97.11% similarity, respectively). However, DNA-DNA hybridization analysis did not reveal any relatedness at the species level. Further differences were apparent in biochemical traits, and fatty acid, quinone and polyamine profiles leading us to conclude that strain SPC-1T (JCM 17498; ITEM 13494) represents a new species of Sphingomonas, for which the name Sphingomonas cynarae sp. nov. is proposed. A component analysis of the exopolysaccharide (named SPC-1T EPS) suggested that it represents a novel type of sphingan containing glucose, rhamnose, mannose and galactose, while glucuronic acid, which is commonly found in sphingans, was not detected.

Table olives are one of the most important traditional fermented vegetables in Southern European countries and their consumption is constantly increasing throughout the world. Today, the industrial production of black table olives is carried out by spontaneous fermentation processes which are not predictable and are strongly influenced by the autochthonous microflora, the physical-chemical conditions, the availability of fermentable substrates and salt content. Evolution of sugars, organic acids, alcohols, mono and polyphenol compounds and volatile compounds associated with the fermentative metabolism of yeasts and bacteria throughout the natural fermentation process of the two Italian olive cultivars Cellina di Nardò and Leccino were determined. A new protocol was developed and applied aimed at the selection of LAB and yeast strains as candidate autochthonous starters for table olive fermentation from Cellina di Nardò and Leccino cultivars.

Plants are ideal bioreactors for the production of macromolecules but transport mechanisms are not fully understood and cannot be easily manipulated. Several attempts to overproduce recombinant proteins or secondary metabolites failed. Because of an independent regulation of the storage compartment, the product may be rapidly degraded or cause self-intoxication. The case of the anti-malarial compound artemisinin produced by Artemisia annua plants is emblematic. The accumulation of artemisinin naturally occurs in the apoplast of glandular trichomes probably involving autophagy and unconventional secretion thus its production by undifferentiated tissues such as cell suspension cultures can be challenging.Here we characterize the subcellular compartmentalization of several known fluorescent markers in protoplasts derived from Artemisia suspension cultures and explore the possibility to modify compartmentalization using a modified SNARE protein as molecular tool to be used in future biotechnological applications. We focused on the observation of the vacuolar organization in vivo and the truncated form of AtSYP51, 51H3, was used to induce a compartment generated by the contribution of membrane from endocytosis and from Endoplasmic Reticulum to Vacuole trafficking.The artificial compartment crossing exocytosis and endocytosis may trap artemisinin stabilizing it until extraction; indeed, it is able to increase total enzymatic activity of a vacuolar marker (RGUSChi), probably increasing its stability. Exploring the 51H3-induced compartment we gained new insights on the function of the SNARE SYP51, recently shown to be an interfering-SNARE, and new hints to engineer eukaryote endomembranes for future biotechnological applications.

Carotenoids are well known for their nutritional properties and health promoting effects representing attractive ingredients to develop innovative functional foods, nutraceutical and pharmaceutical preparations. Pumpkin (Cucurbita spp.) flesh has an intense yellow/orange color owing to the high level of carotenoids, mainly ?-carotene, ?-carotene, ?-cryptoxanthin, lutein and zeaxanthin. There is considerable interest in extracting carotenoids and other bioactives from pumpkin flesh. Extraction procedures able to preserve nutritional and pharmacological properties of carotenoids are essential. Conventional extraction methods, such as organic solvent extraction (CSE), have been used to extract carotenoids from plant material for a long time. In recent years, supercritical carbon dioxide (SC-CO2) extraction has received a great deal of attention because it is a green technology suitable for the extraction of lipophylic molecules and is able to give extracts of high quality and totally free from potentially toxic chemical solvents. Here, we review the results obtained so far on SC-CO2 extraction efficiency and quali-quantitative composition of carotenoids from pumpkin flesh. In particular, we consider the effects of (1) dehydration pre-treatments; (2) extraction parameters (temperature and pressure); the use of water, ethanol and olive oil singularly or in combination as entrainers or pumpkin seeds as co-matrix.

This study investigates the effects of tomato puree fortification (ftp) 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 Daucus carota L. var. atrorubens (Anthocarrot), Vitis vinifera L. fruit skins (Enocolor), Sambucus nigra L. (Elderberry) or Prunus mahaleb L. (Mahaleb) fruits. The consumer acceptance (colour, flavor, taste, visual appearance) was at high level, except for Mahaleb-added extracts. 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, only Anthocarrot-ftp preserved increased phenolic (+53%) content, as well as higher antioxidant capacity (50%). 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.

Watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai) is a well-known refreshing and thirst-quenching typical summer fruit. It has attracted considerable interest for the presence of lycopene and citrulline, biomolecules with undisputed health-promoting effects. Numerous epidemiological studies have revealed the involvement of lycopene in prevention of many degenerative diseases, including arteriosclerosis, diabetes and cancer, likely acting as antioxidant and/or through still poorly understood different molecular mechanisms [1]. Likewise, citrulline, a water-soluble, non-essential amino acid, was recently claimed as having great potential in prevention and treatment of Alzheimer's disease, dementia, erectile dysfunction, high blood pressure, diabetes and, as food supplement, in improving athletic performance [2]. Given the nutritional importance of these bioactive molecules, the preparation of natural concentrated extracts, useful as high-value ingredients for the production of nutraceuticals and/or pharmaceuticals, could represent a viable alternative to the usual production and distribution chain of watermelon and a sustainable process for non-marketable fruits and processing byproducts valorization, opening new opportunities for agri-food industry development.In the last decades the interest towards green, environmentally friendly and safe extraction technologies has strongly increased. Supercritical CO2 extraction has been successfully used to obtain lycopene containing oleoresins from freeze-dried tomato matrices [3,4]. The homogenized flesh of red watermelon (cv. Dumara) peponides was centrifuged to prepare a lycopene rich freeze-dried matrix suitable for supercritical CO2 extraction from the pellet and a citrulline containing powder from the supernatant. The optimal physico-chemical parameters to maximize lycopene extraction yields from the watermelon matrix have been identified. The presence of lipids from watermelon seeds in the matrix has a positive effect on oleoresin extraction. The addition of an oleaginous co-matrix consisting of roughly grinded hazelnut seeds did not further increase the extraction yield. The use of ethanol as co-solvent resulted in a significant increase in lycopene extraction but not of citrulline. The biochemical analyses showed that the extract is a healthy and high added-value source of lycopene and other valuable lipophilic antioxidants useful as bioactive color additive in functional food, nutraceutical, cosmeceutical and pharmaceutical formulations.

Jellyfish are recorded with increasing frequency and magnitude in many coastal areas and several species display biological features comparable to the most popular Asiatic edible jellyfish. The biochemical and antioxidant properties of wild gelatinous biomasses, in terms of nutritional and nutraceutical values, are still largely unexplored. In this paper, three of the most abundant and commonly recorded jellyfish species (Aurelia sp.1, Cotylorhiza tuberculata and Rhizostoma pulmo) in the Mediterranean Sea were subject to investigation. A sequential enzymatic hydrolysis of jellyfish proteins was set up by pepsin and collagenase treatments of jellyfish samples after aqueous or hydroalcoholic protein extraction. The content and composition of proteins, amino acids, phenolics, and fatty acids of the three species were recorded and compared. Protein content (mainly represented by collagen) up to 40% of jellyfish dry weight were found in two of the three jellyfish species (C. tuberculata and R. pulmo), whereas the presence of omega-3 and omega-6 polyunsaturated fatty acids (PUFAs) was significantly higher in the zooxanthellate jellyfish C. tuberculata only. Remarkable antioxidant ability was also recorded from both proteinaceous and non proteinaceous extracts and the hydrolyzed protein fractions in all the three species. The abundance of collagen, peptides and other bioactive molecules make these Mediterranean gelatinous biomasses a largely untapped source of natural compounds of nutraceutical, cosmeceutical and pharmacological interest.

n contrast to the widely accepted consensus of the existence of a single RNA polymerase in bacteria, several actinomycetes have been recently shown to possess two forms of RNA polymerases due theto co-existence of two rpoB paralogs in their genome. However, the biological signi cance of the rpoB duplication is obscure. In this study we have determined the genome sequence of the lipoglycopeptide antibiotic A40926 producer Nonomuraea gerenzanensis ATCC 39727, an actinomycete with a large genome and two rpoB genes, i.e. rpoB(S) (the wild-type gene) and rpoB(R) (the mutant-type gene). We next analyzed the transcriptional and metabolite pro les in the wild-type gene and in two derivative strains over-expressing either rpoB(R) or a mutated form of this gene to explore the physiological role and biotechnological potential of the "mutant-type" RNA polymerase. We show that rpoB(R) controls antibiotic production and a wide range of metabolic adaptive behaviors in response to environmental pH. This may give interesting perspectives also with regard to biotechnological applications.

Lycopene is a carotenoid pigment found in a wide variety of vegetables and fruits. More than six hundred carotenoids have been identified in nature, but only twenty are present in human blood and tissues, including lycopene (the most abundant in plasma), ?-carotene, lutein and zeaxanthin [1]. Several exogenous pigmented antioxidants act on cancer progression and have immune-modulating effects by free radical scavenging activity and/or still unknown mechanisms. However, the controversy recently arisen for some of them requires a careful investigation in relation to their source and synergistic effects [2]. The purpose of this work was: i) to optimize the extraction of lycopene from ripe fruits of tomato, watermelon and GAC by supercritical CO2 in order to obtain natural, solvent-free lycopene containing oleoresins suitable as ingredients for the formulation of innovative functional foods and/or nutraceuticals; ii) to characterize each oleoresin with respect to carotenoids, tocochromanols, lipids, lipophilic and hydrophilic antioxidant activities, micro and macro elements; iii) the preparation of inclusion complexes (ICs) of the oleoresins in food grade ?-cyclodextrins (?-CDs) as hydrophilic nanocarriers [3]; iv) to assess, in-vitro, the effects of administration of each oleoresin/?-CD IC at different concentrations (0.5 to 10 µM) on cell apoptosis and on Interleukin-8 (IL-8) release by A549 human lung adenocarcinoma cells, for 24, 48 and 72 hours.The results showed that all oleoresins represent a healthy and high added-value source of lycopene characterized by: i) the absence of toxic impurities such as solvents, pesticides and heavy metals; ii) a high lycopene bioavailability, increased by the presence of co-extracted natural lipophilic components; iii) a high lipophilic antioxidant activity, improved by synergy with other biomolecules. When administered at a concentration of 10 µM for 72 hours, the three lycopene extracts showed conflicting behaviors: watermelon oleoresin increased cell apoptosis and decreased the release of IL-8, a pro-inflammatory cytokine involved in cancer progression; GAC and tomato oleoresins, instead, decreased cell apoptosis and increased IL-8 release. This discrepancy is likely related to a different lycopene/vitamin E ratio, with watermelon oleoresin showing the highest value among the extracts. Shorter incubation time and lower lycopene concentrations did not affect cell viability and IL-8 release. Further studies are required to assess the possible use of the lycopene containing oleoresin/?-CD ICs in health promoting functional foods and nutraceuticals.

Table olives are one of the most important traditional fermented vegetables in Southern European (Italy, Greece and Spain) countries. In the Greek-style production system, the fruits are placed directly into the brine, thus allowing the natural fermentation to take place. The spontaneous fermentations, that can last 8-12 months, are driven by mixed populations of microorganisms, mainly the epiphytic microbial population of yeasts and lactic acid bacteria (LAB) (Romero et al., 2004). At present, the industrial table olive process is not predictable and depends on the empirical experience of the producers. In order to avoid the unpredictability of the olive spontaneous fermentation, to improve the productive process and to constantly produce high-quality final products, the use of strains of LAB as starter cultures for olive production has been proposed (Sabatini and Marsilioet al., 2008; Panagou et al., 2008; Blana et al., 2014). However, in the last years, the importance and the potential applications of yeasts as starters for table olive processing has been recognized (Arroyo-López et al., 2008, 2012; Bevilacqua et al., 2012). Objectives: In the present work, we have studied the main physical, chemical and aromatic parameters of natural fermentations of Cellina di Nardò, Leccino, Kalamata and Conservolea table olives in order to determine chemical descriptors correlated to microbiological activities and the dynamics of microorganisms in order to select LAB and yeast strains as candidate autochthonous starter cultures. Conclusions: The identified chemical descriptors can be suitable to follow the trend and to control the outcome of the fermentation and a new protocol aimed to the selection of LAB and yeast strains as candidates autochthonous starters has been developed and applied (Bleve et al. 2014 a, b). Selected microbial starters have been successfully used to ferment olives in pilot and industrial-scale and a new method for table olive production has been set up (Bleve et al. 2103). The use of selected autochthonous starter cultures produced fermented table olives with improved organoleptic, sensorial and nutritional characteristics.

Lycopene is an antioxidant carotenoid pigment found in a wide variety of vegetables and fruits, together with other lipophilic phytochemicals. Antioxidants have different effects on cancer progression. Non-small cell lung cancer (NSCLC) accounts for >80% of all lung cancers. Inflammation is a key event in cancer development. This study aims to evaluate in vitro the effects of administration of three different oleoresins containing lycopene on human NSCLC cell line, A549. The oleoresins, obtained by the supercritical CO2 green extraction technology from watermelon (Lyc W), GAC (Lyc G) and tomato (Lyc T), were chlatrated in ?-cyclodextrins (?-CD) in order to improve their stability and promote the dispersion of the lipophilic compounds in the cell-culture aqueous medium. They were tested at different concentrations (from 0.5 to 10 µM lycopene) on A549 by the Cell Proliferation Assay (MTS) for 24 hrs, Annexin V-FITC/PI apoptosis test and ELISA for Interleukin-8 (IL-8) release, for 48 and 72 hrs. The oleoresins were characterized by a different composition in carotenoids and tocochromanols, with Lyc W showing the highest lycopene/tocochromanol ratio. At 10 µM, the three extracts showed conflycting behaviors: Lyc W increased cell apoptosis (p=0.01) and decreased cell proliferation (p=0.009) and IL-8 release (p=0.03), whereas Lyc G and Lyc T decreased cell apoptosis(p=0.01 and p=0.04, respectively) and increased cell proliferation (p=0.01 and p=0.04, respectively) and IL-8 release (both p=0.01). Furthermore, at 3 µM, Lyc G and Lyc T decreased cell apoptosis (p=0.009 and p=0.01, respectively). In Lyc W the highest amount of lycopene is able to counteract and revert the survival effect of tocochromanols, supporting the importance to evaluate the real effect of antioxidant supplementation on lung cancer which not only may have no anticancer benefits but even increase cancer aggressivity.

Agri-food by-products produced during handling and processing of fruits and vegetables, represent a major waste disposal problem for industry, but they are also promising sources of bioactive compounds that can be recovered and used as valuable substances to develop new enriched food, such as pasta and bakery products. Tarallini are a typical Italian bakery products, that has currently become very popular worldwide as a savory snack or bread substitute. These products can be considered an optimal, cheap and easy-to-use carrier of phytochemicals.Olive fruit is an excellent natural source of unsaturated fatty acids, as well as other nutritionally important health- promoting bioactive compounds [1]. The modern two-phase technology of the Leopard series for olive oil industrial extraction developed by the Pieralisi Group (Pieralisi Maip S.p.A., Jesi, AN, Italy) generates large quantities of a novel by-product (olive paste - OP) made up of the partially defatted wet drupe pulp without any traces of the kernel. OP can be incorporated into wheat based products [2].In this work the biotechnological aptitude of selected microorganisms, yeast and Lactic Acid Bacteria from different sources, were tested to transform OP in a new fermented product. Furthermore, some bioactive compounds, such as polyphenols, triterpenic acids, phytosterols, squalene, tocopherols and carotenoids, of fermented OP obtained from black olives belonging to Leccino and Cellina di Nardò cultivars, were characterized. Finally, new types of tarallini were produced by adding 10% of fermented and non-fermented OP to dough. The levels of bioactive compounds were compared with conventional control tarallini produced with the same semolina but without supplementations.The polyphenols content of olive paste, found in the olive paste Leccino (OPL) was 7.2 mg/g fresh (FW) while for olive paste Cellina di Nardò (OPC) the concentration was 18.1 mg/g FW. The fermentation of OP produces a decrease of phenolic concentration 20% for OPL and 16% for OPC. Among the oleuropein derivatives, 3,4- DHPEA-EDA, showed the higher reductions corresponding to the increase in their hydrolysis products (hydroxytyrosol), due to the enzymatic activity of the strains used for the fermentation of the OP [3].Total triterpenic acid content was about 1.5 and 1.7 mg/g fresh weight in OPL and OPC respectively. These values did not differ significantly compared with the respective fermented OP. The total phytosterols and squalene content ranged between 1.7 mg/g FW (OPC) and 1.9 mg/g FW (OPL) and between 29.10 ?g/g FW (OPL) and 38.95 ?g/g FW (OPC), respectively. After fermentation, total phytosterols content was reduced from 53% to 49% whereas squalene level was reduced from 47% to 23% compared with not fermented OP. The content of tocochromanol and carotenoid did not differ significantly after fermentation process, their content ranged from 22.64 ?g/g FW (fermented OPL) to 24.94 ?g/g FW (not fermented OPL) for tocochrom

During recent years food industries generally produce a large volume of wastes both solid and liquid, representing a disposal and potential environmental pollution problem.Objective: The goal of the study was to optimize, from both sensory and nutritional points of view, the formulation of durum wheat spaghetti enriched with an olive oil industrial by-product,indicated as olive paste. Methods: Three consecutive steps were carried out. In the first one, the olive paste was air-dried at low temperature, milled to record olive paste flour and properly analyzedfor its biochemical composition. In the second step, the olive paste flour was added to the pasta dough at 10% and 15% (w/w). In the last step, different concentrations of transglutaminase wereadded to enriched pasta (10% olive paste) to further improve the quality. Sensory properties and nutritional content of enriched and control pasta were properly measured. Results: Spaghetti with10% olive paste flour and 0.6% transglutaminase were considered acceptable to the sensory panel test. Nutritional analyses showed that addition of 10% olive paste flour to pasta considerably increased content of flavonoids and total polyphenols. Conclusions: The proper addition of olive paste flour and transglutaminase for pasta enrichment could represent a starting point to valorize olive oil industrial by-products and produce new healthy food products.

Phenolic compounds are a widespread group of substances in the plant kingdom, but their dietary intake greatly varies depending on the type and quantity of vegetable foods consumed. Interest in plant polyphenols has focused on their favourable effects on human health, arising from their antioxidant activity and capacity to protect critical macromolecules (such as chromosomal DNA, structural proteins and enzymes, low-density lipoproteins, and membrane lipids) from damages induced by active species of oxygen. Phenolic substances are present also in wheat, with ferulic acid present in the highest amount. Various studies analysed the polyphenol content of soft, medium and hard wheat (Triticum aestivum L.). Marginal attention has been devoted to durum wheat (Triticum turgidum L. spp. turgidum var. durum), the essential raw material to produce high quality pasta, one of the basic foods in the Italian diet. The aim of this work has been to evaluate the variability of total phenolic content (TPC) in Italian durum wheat, in view of selecting the best materials to produce phenolic extracts for functional pasta enrichment. A set of 20 cultivars was considered, chosen among the most cultivated ones. The determination of TPC of the free phenolic fraction, spectrophotometrically assessed after Folin-Ciocalteau reaction, showed levels ranging from 1.28 to 1.94 mg ferulic acid equivalent (FAE)/g wholemeal. The TPC was significantly correlated to the polyphenol oxidase activity of the examined cultivars.A subset of 5 cultivars was subjected to detailed survey of phenolic acids in whole flour. Three separate extractions were carried out to determine the composition and amount of soluble free, soluble conjugated, and bound phenolic acids. Two different mesh size (60 mesh or 0.251 mm; and 10 mesh or 2 mm) were used for milling whole grain samples to evaluate the extraction efficiency of phenolic acids depending on the particle size of whole flour samples. The evaluation of free, conjugated and bound extracts by the means of HPLC-DAD analysis lead to the characterisation of each fraction in terms of composition and amount of major phenolic acids. Sinapic acid resulted the most abundant compound present in the bound fraction, ranging from 335 and 438 ug/g dm, while o-coumaric acid resulted the major component of the conjugated fraction, ranging from 72 and 80 ug/g dm. Other phenolic acids were detected both in the bound and conjugated fractions, like ferulic, 2,4-dihydroxybenzoic, syringic and vanillic acids. The free fraction showed the smallest contribution to the total phenolic acid content (<0.5%), sinapic acid being the most abundant, followed by syring, ferulic and o-coumaric acid respectively.

Volatile metabolites from mold contamination have been proposed for the early identification of toxigenic fungi to prevent toxicological risks, but there are no such data available for Fusarium poae. F. poae is one of the species complexes involved in Fusarium head blight, a cereal disease that results in significant yield losses and quality reductions. The identification of volatile organic compounds associated with F. poae metabolism could provide good markers to indicate early fungal contamination. To this aim, we evaluated the volatile profile of healthy and F. poae-infected durum wheat kernels by SPME-GC/MS analysis. The production of volatile metabolites was monitored for seven days, and the time course analysis of key volatiles was determined. A total of 29 volatile markers were selected among the detected compounds, and multivariate analysis was applied to establish the relationship between potential volatile markers and fungal contamination. A range of volatile compounds, including alcohols, ketones, esters, furans and aromatics, were identified, both in contaminated and in healthy kernels. However, the overall volatile profile of infected samples and controls differed, indicating that the whole volatile profile, rather than individual volatile compounds, could be used to identify F. poae contamination of durum wheat grains.

The invention relates to a method for table olive fermentation comprising the steps of: a. soaking the olives in brine;b. inoculating the product obtained in step a. with a yeast culture;c. incubating the product obtained in step b. in order to perform the alcoholic fermentation; d. inoculating the fermented product obtained in step c. using a Lactic Acid Bacterium (LAB) culture;e. incubating the product obtained in step d. to in order to perform the lactic fermentation.Procedures for the selection of starter cultures and their use in fermentation of two cultivars of table olives are described. Some claims are directed to specific starter cultures and the uses thereof for preparing fermented table olives.