Paolo Rapisarda

Fruit quality and bioactive compounds relevant to human health of sweet cherry (Prunus avium L.) cultivars grown in Italy

The fruit quality characteristics, phenolic compounds and antioxidant capacities of 24 sweet cherry (Prunus avium L.) cultivars grown on the mountainsides of the Etna volcano (Sicily, Italy) were evaluated. High-performance liquid chromatographic methods were used to identify and quantify sugars, organic acids and phenolics. A total of seven phenolic compounds were characterised as hydroxycinnamic acid derivatives (neochlorogenic acid, p-coumaroylquinic acid and chlorogenic acid) and anthocyanins (cyanidin 3-glucoside, cyanidin 3-rutinoside, pelargonidin 3-rutinoside and peonidin 3-rutinoside). The total anthocyanin content ranged from 6.21 to 94.20mg cyanidin 3-glucoside equivalents/100g fresh weight (FW), while the total phenol content ranged from 84.96 to 162.21mg gallic acid equivalents/100g FW. The oxygen radical absorbance capacity (ORAC) assay indicated that fruit of all genotypes possessed considerable antioxidant activity. The high level of phenolic compounds and antioxidant capacity of some sweet cherry fruits implied that they might be sources of bioactive compounds that are relevant to human health.

Antiinflammatory effects of a red orange extract in human keratinocytes treated with interferon-gamma and histamine.

Red oranges are an important component of the so‐called Mediterranean diet and they have been used by traditional medicine for their health protective properties, particularly to heal sore throat and cough, suggesting an interesting antiinflammatory activity. The purpose of this study was to evaluate the antiinflammatory activity of a red orange (Citrus sinensis varieties: Moro, Tarocco, Sanguinello) complex (ROC), characterized by high levels of anthocyanins, flavanones, hydroxycinnamic acids and ascorbic acid, on the human keratinocyte line NCTC 2544 exposed to interferon‐gamma (IFN‐γ) and histamine. The expression of immunomodulatory membrane molecules such as inter‐cellular adhesion molecule‐1 (ICAM‐1) by Western blot analysis, and the release of chemokines such as monocyte chemoattractant protein‐1 (MCP‐1) and interleukin‐8 (IL‐8) through ELISA kits, were determined. ICAM‐1 modulates the permanence and activation of T lymphocytes in the epidermis. MCP‐1 is a specific chemoattractant for monocytes and dendritic cells. IL‐8 is important for the recruitment of both neutrophils and T lymphocytes. Addition of ROC at different concentrations together with IFN‐γ and histamine induced a dose‐dependent inhibition of ICAM‐1 expression and MCP‐1 and IL‐8 release. ROC shows interesting antiinflammatory properties in human keratinocyte cells NCTC 2544. This natural complex could have a topical employment and mitigate the consequences of some skin pathologies. Copyright

Chemical Characterization of Different Sumac and Pomegranate Extracts Effective against Botrytis cinerea Rots.

Pomegranate (Punica granatum L.) peel and sumac (Rhus coriaria L.) fruit and leaf extracts were chemically characterized and their ability to inhibit table grape (cv. Italia) rots caused by Botrytis cinerea was evaluated on artificially inoculated berries. Different extraction methods were applied and extracts were characterized through Ultra Fast High Performance Liquid Chromatography coupled to Photodiode array detector and Electrospray ionization Mass spectrometer (UPLC-PDA-ESI/MSn) for their phenol and anthocyanin contents. The concentrated pomegranate peel extract (PGE-C) was the richest in phenols (66.97 g gallic acid equivalents/kg) while the concentrated sumac extract from fruits (SUF-C) showed the highest anthocyanin amount (171.96 mg cyanidin 3-glucoside equivalents/kg). Both phenolic and anthocyanin profile of pomegranate and sumac extracts were quite different: pomegranate extract was rich in cyanidin 3-glucoside, pelargonidin 3-glucoside and ellagic acid derivatives, while sumac extract was characterized by 7-methyl-cyanidin 3-galactoside and gallic acid derivatives. The concentrated extracts from both pomegranate peel and sumac leaves significantly reduced the development of Botrytis rots. In particular, the extract from pomegranate peel completely inhibited the pathogen at different intervals of time (0, 12, and 24 h) between treatment and pathogen inoculation on fruits maintained at 22–24 °C and high relative humidity (RH). This extract may represent a valuable alternative to control postharvest fungal rots in view of its high efficacy because of the low cost of pomegranate peel, which is a waste product of processing factories.

Protective effects of a standardised red orange extract on air pollution-induced oxidative damage in traffic police officers.

Several pathological conditions have all been associated with a higher release of atmospheric pollutants. There is growing evidence that oxidative stress may represent one of the agents involved in the initiation and/or progression of many of these pathologies. The aim of the present study was to evaluate the effects of short-term dietary supplementation with a standardised red orange extract (ROC) on a group of traffic police officers exposed to traffic exhaust pollution and cigarette smoking, by measuring some noninvasive biomarkers of oxidative stress. At the beginning of the study, all the groups showed similar serum lipid hydroperoxide levels, but traffic officers showed lower serum concentrations of thiol (SH) groups; furthermore, the frequency of spontaneous sister chromatide exchanges (SCEs) in peripheral lymphocytes was increased by smoking (but not by pollution exposure alone) at a higher degree in subjects exposed to traffic pollution. After 1 month of ROC administration, serum lipid hydroperoxide levels decreased only in all non-smoking subjects; furthermore, SH group levels measured in traffic officers appeared restored to normal values observed in the respective controls. Finally, the increase in SCE frequency induced by smoking was reduced by treatment with ROC especially in traffic officers. Our study suggests that ROC supplementation could be useful to minimise the detrimental effects caused by exposure to air pollution and smoking.

Essential oil profiles of new Citrus hybrids, a tool for genetic citrus improvement

The essential oil composition of new Citrus hybrids obtained from the Citrus genetic improvement programs at the CRA-Centro di Ricerche per l’Agrumicoltura e le Colture Mediterranee of Acireale, Italy (CRA-ACM), were used to evaluate the fruit quality and to identify new valuable flavor components. The common objective of genetic improvement programs is to generate Citrus cultivars with a higher resistance to disease and to biotic and abiotic stresses, as well as seedless and easy to peel fruits with optimal size and shape and with new organoleptic characteristics. The chemical characterization of the volatile components in the essential oils from the peel of new Citrus hybrids were determined by gas chromatography combined with mass spectrometry (GC-MS). The fruits studied were respectively: seven hybrids (P1–P7) coming from crossing a diploid clementine with a tetraploid grapefruit; a new hybrid with lemon-like character obtained by crossing a diploid clementine with a tetraploid lemon; two pigmented triploid hybrids obtained by crossing a diploid clementine with a tetraploid ‘Tarocco’ orange (‘A146’ and ‘C1867’); a new mandarin hybrid named ‘Cami’; four pigmented hybrids whose parents were clementine and ‘Moro’ orange (‘OMO 15’, ‘OMO 31’, ‘OMO 6’ and ‘OMO 12’); and finally, a red-fleshed hybrid (Amoa-8) obtained from ‘Moro’ orange and ‘Avana’ mandarin.

Oxygen radical scavenging capacity of phenolic and non-phenolic compounds in red and white wines

The aim of the present study was the evaluation of the antioxidant content in phenolic and non-phenolic extracts of ten wine samples, trying to elucidate the potential role of unusual antioxidant compounds. Samples of wines processed from red and white grapes (Vitis vinifera L.), deprived of the volatile fraction at low temperature and buffered at physiological pH, were fractionated by C18 into two fractions: FR1 and FR2. Non-phenolics, such as tartaric, malic, lactic, and succinic acids; glucose; fructose; and glycerin were mainly found in FR1, while polyphenols were present exclusively in FR2. Peroxyl radical quenching was assayed by the ORAC method, while superoxide and hydroxyl radical scavenging activity were assayed by electron paramagnetic resonance. In the ORAC and superoxide assays, most of the activity was found in FR2, while in hydroxyl radical assay, the activity was found in FR1. Model solutions were used to attribute a role to the single compounds in the evaluation of wine’s ROS scavenging capacity: the ORAC and superoxide anion scavenging effects were mainly attributed to the polyphenols, averaging 94.8%, with some contribution from glycerin, particularly in white wines. Unexpectedly, the main chemical responsible for hydroxyl radical scavenging activity was glycerin (56.1%), with the polyphenols scavenging at 18.1%.

Involvement of inducible nitric oxide synthase and cyclooxygenase-2 in the anti-inflammatory effects of a red orange extract in human chondrocytes.

In the present study, a complex of compounds (red orange complex, ROC), obtained from three red orange varieties (Citrus sinensis varieties: Moro, Tarocco and Sanguinello), containing cyanidin glycosides, hydroxycinnamic acids, flavanone glycosides and ascorbic acid, was screened to discover new lead compounds in the suppression of the production of key molecules released during inflammatory events in interleukin-1β (IL-β) stimulated human primary chondrocytes. The expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX)-2 and intercellular adhesion molecule-1 (ICAM-1), and the release of nitric oxide, prostaglandin (PG)E2 and interleukin-8 (IL-8) were determined. Indomethacin was used as an anti-inflammatory drug reference. ROC acts as a potent inhibitor of iNOS and COX-2 gene expression while also suppressing the production of PGE2 and nitrite in human chondrocytes. In addition, ROC induces a significant decrease in ICAM expression and IL-8 release. These findings suggest that ROC exerts anti-inflammatory effects probably through the suppression of COX-2 and iNOS expression.

Methods used to evaluate the peroxyl (ROO·) radical scavenging capacities of four common antioxidants

Different methods that are used to measure peroxyl radical (ROO·) scavenging capacity were used in model systems containing four common antioxidant compounds: ascorbic acid, cysteine, gallic acid and Trolox, a water-soluble vitamin E analogue. Five tests were performed, with differences in the method of peroxyl radical generation (lipoxygenase, azo-compound thermal degradation and spontaneous oxidation from a natural source of fatty acids) and in the addition of probes such as linoleic acid, crocin, fluorescein and electron paramagnetic resonance (EPR) by spin-trapping with α-phenyl-N-tert-butyl-nitrone. Significant differences were found in antioxidant reactivity, evaluated by dose–effect semi-logarithmic plots and EC50 values, and a possible explanation for these differences was explored. In assays in which ROO· was generated by the azo-compound or spontaneous oxidation, gallic acid was the most powerful antioxidant. When lipoxygenase was used to produce ROO·, ascorbic acid had the highest scavenging index, while cysteine and gallic acid had low levels of activity. Another difference was identified regarding the molecular probe: ascorbic acid and Trolox were the least reactive in the EPR system and azo-bis system with fluorescein, while in assays with crocin, these compounds had intermediate activities. Ascorbic acid and Trolox were the best antioxidants in an enzymatic-linoleic acid system, with a reciprocal ratio of 6:1, while adding crocin to the same system changed the reciprocal ratio to 2:1. A literature examination indicated that out of the antioxidant compounds tested, cysteine has the highest bond dissociation enthalpy, and this compound showed, as expected, low scavenging activity in three of the five tests in this study.

Evaluation of a Pomegranate Peel Extract (PGE) as Alternative Mean to Control Olive Anthracnose

Olive anthracnose is caused by different species of Colletotrichum spp. and may be regarded as the most damaging disease of olive fruit worldwide, greatly affecting quality and quantity of the productions. A pomegranate peel extract (PGE) proved very effective in controlling the disease. The extract had a strong in vitro fungicidal activity against Colletotrichum acutatum sensu stricto, was very effective in both preventive and curative trials with artificially inoculated fruit, and induced resistance in treated olive tissues. In field trials, PGE was significantly more effective than copper, which is traditionally used to control the disease. The highest level of protection was achieved by applying the extract in the early ascending phase of the disease outbreaks because natural rots were completely inhibited with PGE at 12 g/liter and were reduced by 98.6 and by 93.0% on plants treated with PGE at 6 and 3 g/liter, respectively. Two treatments carried out 30 and 15 days before the expected epidemic outbreak reduced the incidence of the disease by 77.6, 57.0, and 51.8%, depending on the PGE concentration. The analysis of epiphytic populations showed a strong antimicrobial activity of PGE, which sharply reduced both fungal and bacterial populations. Because PGE was obtained from a natural matrix using safe chemicals and did not have any apparent phytotoxic effect on treated olive fruit, it may be regarded as a safe and effective natural antifungal preparation to control olive anthracnose and improve olive productions.

An Alginate/Cyclodextrin Spray Drying Matrix to Improve Shelf Life and Antioxidant Efficiency of a Blood Orange By-Product Extract Rich in Polyphenols: MMPs Inhibition and Antiglycation Activity in Dysmetabolic Diseases

Alginate and β-cyclodextrin were used to produce easily dosable and spray-dried microsystems of a dried blood orange extract with antidysmetabolic properties, obtained from a by-product fluid extract. The spray-dried applied conditions were able to obtain a concentrate dried extract without the loss of AOA and with TPC and TMA values of 35–40% higher than that of the starting material. They were also effective in producing microparticles with 80–100% of encapsulation efficiency. The 2% sodium alginate was capable of improving the extract shelf life, while the beta-cyclodextrin (1 : 1 molar ratio with dried extract) prolonged the extract antioxidant efficiency by 6 hours. The good inhibition effect of the dried extract on the AGE formation and the MMP-2 and MMP-9 activity is presumably due to a synergic effect exerted by both anthocyanin and bioflavonoid extract compounds and was improved by the use of alginate and cyclodextrin.