Almond [Prunus dulcis (Mill.) DA Webb] Processing Residual Hull as a New Source of Bioactive Compounds: Phytochemical Composition, Radical Scavenging and Antimicrobial Activities of Extracts from Italian Cultivars ('Tuono', 'Pizzuta', 'Romana').

In this study we developed a new extract, by the use of conventional solid-solvent extraction and a food-grade hydroalcoholic solvent, rich in phenolic and triterpenoid components from almon hull to be employed as functional ingredient in food, pharma and cosmetic sectors. Two autochthonous Sicilian cultivars ('Pizzuta' and 'Romana') and an Apulian modern cultivar ('Tuono') have been tested for the production of the extract. Results showed that the two Sicilian varieties, and in particular the 'Romana' one, present the best characteristics to obtain extracts rich in triterpenoids and hydroxycinnamic acids, useful for the production of nutraceutical supplements. About triterpenoids, the performance of the hydroalcoholic extraction process allowed to never go below 46% of recovery for 'Pizzuta' samples, with significantly higher percentages of recovery for 'Tuono' and 'Romana' extracts (62.61% and 73.13%, respectively) while hydroxycinnamic acids were recovered at higher recovery rate (84%, 89% and 88% for 'Pizzuta', 'Romana' and 'Tuono' extracts, respectively). Invitro antioxidant and antimicrobial activities exerted by the extracts showed promising results with P. aeruginosa being the most affected strain, inhibited up to the 1/8 dilution with 'Romana' extract. All the three tested extracts exerted an antimicrobial action up to 1/4 dilutions but 'Romana' and 'Pizzuta' extracts always showed the greatest efficacy.

Distribution, Antioxidant Capacity, Bioavailability and Biological Properties of Anthocyanin Pigments in Blood Oranges and Other Citrus Species.

Anthocyanins are natural pigments that give a red, purple, and blue color to many plant, flower, fruit, and vegetable species. Their presence within the genus Citrus was first reported in 1916, and it is well-known that the red color of the flesh and rind of blood (red or pigmented) oranges (Citrus sinensis L. Osbeck) is due to the presence of anthocyanins. They are also present in the young shoots, flowers, and peel of lemon (Citrus limon (L.) Burm. f.), citron (Citrus medica L.), and other citrus species. Since then, the scientific community has expressed increasing interest in studying their profile and distribution, with many published studies focused on the quali-quantitative pattern in the different vegetative tissues belonging to the genus Citrus. Moreover, with the discovery of their relevant antioxidant activity, owing to their ability to capture free radicals, much research has been performed in the last two decades on their radical scavenging power, in vitro and in vivo biological properties, and anticarcinogenic capacity, also focusing attention on their bioavailability for humans. The present work is intended as a comprehensive review of the advances in scientific research on anthocyanin pigments naturally occurring within the genus Citrus, including their natural distribution, antioxidant capacity, bioavailability, and biological value and properties. The scientific evidence herein reported can be used to further increase the knowledge of this class of compounds and represents a valuable and comprehensive contribution to promoting anthocyanin-rich citrus fruit consumption as a healthy dietary habit.

Multielemental, Nutritional, and Proteomic Characterization of Different Lupinus spp. Genotypes: A Source of Nutrients for Dietary Use.

Among grain pulses, lupins have recently gained considerable interest for a number of attractive nutritional attributes relating to their high protein and dietary fiber and negligible starch contents. The seeds of Lupinus albus (cv. Multitalia and Luxor, and the Modica ecotype); L. luteus (cv. Dukat, Mister, and Taper); and L. angustifolius (cv. Sonet) analyzed in this study were deposited within the germplasm collection of the Research Centre for Cereal and Industrial Crops of Acireale and were sowed in East Sicily in 2013/14. The collected seeds were analyzed for their multielemental micro- and macronutrient profiles, resulting in a wide variability between genotypes. Lupin seed flour samples were subjected to a defatting process using supercritical CO2, with oil yields dependent on the species and genotype. We determined the fatty acid profile and tocopherol content of the lupin oil samples, finding that the total saturated fatty acid quantities of different samples were very close, and the total tocopherol content was about 1500.00 µg/g FW. The proteomic analysis of the defatted lupin seed flours showed substantial equivalence between the cultivars of the same species of Lupinus albus and L. luteus. Moreover, the L. angustifolius proteome map showed the presence of additional spots in comparison to L. albus, corresponding to α-conglutins. Lupin, in addition to being a good source of mineral elements, also contributes vitamin E and, thanks to the very high content of gamma-tocopherols, demonstrates powerful antioxidant activity.

Prebiotic effects of citrus pectic oligosaccharides.

Citrus peel wastes, one of the major by-products of the agri-food industry, are a source of value-added compounds. In this work, a commercial pectin (PEC1) and one obtained by direct extraction from citrus fibre (PEC2), were hydrolysed and ultrafiltered at different cut-off (100, 50 and 30 kDa) and then tested in broth with four different probiotic strains: Lacticaseibacillus paracasei, Limosilactobacillus fermentum, Lacticaseibacillus rhamnosus and Bifidobacterium animalis subsp. paracasei. In addition, the probiotic strains selected for their growth rate with the fractions of the two pectins were tested for tolerance to different pH values and bile salts. This study demonstrated that POS2 (product from the enzymatic hydrolysis by PEC2) showed greater prebiotic activity than POS1 (product from the enzymatic hydrolysis by PEC1). The results confirmed that citrus pectic oligosaccharides could be used as emerging prebiotics with improved properties due to their antimicrobial and modulating-microbiota ability.

Bioactive Compounds from Lemon (Citrus limon) Extract Overcome TNF-α-Induced Insulin Resistance in Cultured Adipocytes.

The consumption of plant-based food is important for health promotion, especially regarding the prevention and management of chronic diseases such as diabetes. We investigated the effects of a lemon extract (LE), containing ≥20.0% total flavanones and ≥1.0% total hydroxycinnamic acids, on insulin signaling in murine 3T3-L1 adipocytes treated with TNF-α, which was used to mimic in vitro the insulin resistance condition that characterizes diabetes mellitus. Our results showed LE increased PPARγ, GLUT4 and DGAT-1 levels, demonstrating the potential of this lemon extract in the management of insulin resistance conditions associated with TNF-α pathway activation. LE treatment further decreased the release of interleukin 6 (IL-6) and restored triglyceride synthesis, which is the main feature of a healthy adipocyte.

A Standardized Extract Prepared from Red Orange and Lemon Wastes Blocks High-Fat Diet-Induced Hyperglycemia and Hyperlipidemia in Mice.

Citrus fruits are a rich source of high-value bioactive compounds and their consumption has been associated with beneficial effects on human health. Red (blood) oranges (Citrus sinensis L. Osbeck) are particularly rich in anthocyanins (95% of which are represented by cyanidin-3-glucoside and cyanidin-3-6″-malonyl-glucoside), flavanones (hesperidin, narirutin, and didymin), and hydroxycinnamic acids (caffeic acid, coumaric acid, sinapic, and ferulic acid). Lemon fruit (Citrus limon) is also rich in flavanones (eriocitrin, hesperidin, and diosmin) and other polyphenols. All of these compounds are believed to play a very important role as dietary antioxidants due to their ability to scavenge free radicals. A standardized powder extract, red orange and lemon extract (RLE), was obtained by properly mixing anthocyanins and other polyphenols recovered from red orange processing waste with eriocitrin and other flavanones recovered from lemon peel by a patented extraction process. RLE was used for in vivo assays aimed at testing a potential beneficial effect on glucose and lipid metabolism. In vivo experiments performed on male CD1 mice fed with a high-fat diet showed that an 8-week treatment with RLE was able to induce a significant reduction in glucose, cholesterol and triglycerides levels in the blood, with positive effects on regulation of hyperglycemia and lipid metabolism, thus suggesting a potential use of this new phytoextract for nutraceutical purposes.

Physico-chemical and multielemental traits of anaerobic digestate from Mediterranean agro-industrial wastes and assessment as fertiliser for citrus nurseries.

Previous researches have demonstrated the bioenergetic potential of agri-food Mediterranean wastes showing that anaerobic co-digestion is a valuable solution for Mediterranean areas. This implies a great interest for anaerobic digestates use in agriculture to replace fertilizers. The present study aimed at: i) producing knowledge on continuous anaerobic co-digestion of feedstock mixture composed by different Mediterranean agri-food wastes in terms of multielemental characterization and ii) assessing the agronomic value of industrial anaerobic digestate (AD) based on the potential as fertiliser in nursery condition for the citrus seedlings. Results have demonstrated that agro-industrial biomasses have great potentiality to be converted by anaerobic digestion in biofertilizer to be used in citrus nurseries as sustainable alternative to mineral fertilisers. Multielemental traits of the tested AD were valuable in terms of nutritional supply for the growth and development of the plant. AD was useful to replace the mineral fertilizers in terms of total N content (10.81 ± 0.32 %TS) and organic matter (43.32 ± 0.80 %TS). The seedlings nutritive status showed that no need for supplemental of nutrients was requested. Volkamer lemon highly benefited from the administration of liquid digestate, increasing the total chlorophyll level (2.97 ± 0.31 mg g-1 FW) presumably due to the higher ammonium content of the AD (59 ± 0.08 %TKN). Besides providing useful tools for citrus nurseries for conceiving new sustainable fertilization strategies, this study is a starting point for further in-depth works on physiological status and traits of citrus plants fertilized by using agro-industrial anaerobic digestate.

Microbial Application to Improve Olive Mill Wastewater Phenolic Extracts.

Olive mill wastewater (OMW) contains valuable and interesting bioactive compounds, among which is hydroxytyrosol, which is characterized by a remarkable antioxidant activity. Due to the health claims related to olive polyphenols, the aim of this study was to obtain an extract from OMW with an increased level of hydroxytyrosol by means of microbial enzymatic activity. For this purpose, four commercial adsorbent resins were selected and tested. The beta-glucosidase and esterase activity of strains of Wickerhamomyces anomalus, Lactiplantibacillus plantarum, and Saccharomyces cerevisiae were also investigated and compared to those of a commercial enzyme and an Aspergillus niger strain. The W. anomalus strain showed the best enzymatic performances. The SP207 resin showed the best efficiency in selective recovery of hydroxytyrosol, tyrosol, oleuropein, and total phenols. The bioconversion test of the OMW extract was assessed by using both culture broths and pellets of the tested strains. The results demonstrated that the pellets of W. anomalus and L. plantarum were the most effective in hydroxytyrosol increasing in phenolic extract. The interesting results suggest the possibility to study new formulations of OMW phenolic extracts with multifunctional microorganisms.

Evaluation of lipid and cholesterol-lowering effect of bioflavonoids from bergamot extract.

Several natural products have been reported to be involved in the suppression of adipogenesis. In this study, we reveal that a bergamot extract (BE) decreased the accumulation of intracellular lipids in murine pre-adipocytes 3T3-L1 cells during adipogenic differentiation. Both the inhibition of HMG-CoA reductase activity and the differentiation and proliferation of adipocytes could be used as a strategy for the treatment and prevention of obesity. The results of this study show a reduction of HMG-CoA activity and of lipid droplet accumulation in the presence of the BE, suggesting the potential of BE as an anti-adipogenic agent to lower the content of cholesterol and body fat and prevent a gain in body weight. Moreover, BE as the result of high percentages of flavonoid compounds such as neoriocitrin, naringin and neohesperidin, the main flavonoids contained in BE, led to significant inhibition of DPPH free radical, demonstrating a strong radical scavenging activity.

A new standardized phytoextract from red orange and lemon wastes (red orange and lemon extract) reduces basophil degranulation and activation.

Citrus fruits are rich sources of bioactive compounds and their consumption is associated to health-promoting effects. Citrus processing wastes contain bioflavonoids and other high added value compounds. The aim of this study was to evaluate the antiallergic properties of a new phytoextract obtained by citrus wastes and peels. Blood orange and lemon processing wastes were used to produce a Red orange and Lemon Extract (RLE). Blood samples from 30 allergic donors were collected and used to evaluate the basophil activation (CD203c) and degranulation (CD63) by stimulation trough allergen with and without the RLE. Reduced basophil expression of CD203c and CD63 were observed in RLE + Allergen treated samples, with -20.21% of CD203c expression (p < 0.0001) and -54.11% of CD63 expression (p < 0.0001), compared to Allergen treated samples. The RLE evidenced a good antiallergic activity, mainly acting on basophils degranulation, and therefore reducing the key event of pro-inflammatory mediators release after allergic stimuli.

In vitro effects of bioflavonoids rich lemon extract on pre-adipocyte differentiation.

Lemon fruit is a source of bioactive compounds, which has many beneficial effects on health. Obesity is characterized by over-accumulation of adipose tissue as a result of increased adipocyte size and number. Adipogenesis is mediated and assisted by various transcription factors that induce lipid-metabolizing enzymes followed by an increase of perilipin expression and lipid droplets generation. Here, we evaluate the effect of lemon extract (LE) as radical scavenger and the consequent regulation of adipocyte differentiation and lipid accumulation. 3T3-L1 murine pre-adipocytes were differentiated and treated with different LE concentrations. The high percentages of flavonoid contained in LE led to a significant inhibition of DPPH radical and reactive oxygen species, demonstrating a strong radical scavenger activity. Treatment of 3T3-cells with LE showed a significant decrease of perilipin expression, subsequently confirmed by the reduction of lipid droplet accumulation, resulting from Oil Red O Staining and by the downregulation of PPARγ and DGAT-1mRNA.

Development of Durum Wheat Breads Low in Sodium Using a Natural Low-Sodium Sea Salt.

Durum wheat is widespread in the Mediterranean area, mainly in southern Italy, where traditional durum wheat breadmaking is consolidated. Bread is often prepared by adding a lot of salt to the dough. However, evidence suggests that excessive salt in a diet is a disease risk factor. The aim of this work is to study the effect of a natural low-sodium sea salt (Saltwell®) on bread-quality parameters and shelf-life. Bread samples were prepared using different levels of traditional sea salt and Saltwell®. The loaves were packaged in modified atmosphere conditions (MAPs) and monitored over 90 days of storage. No significant differences (p ≤ 0.05) were found in specific volumes and bread yield between the breads and over storage times, regardless of the type and quantity of salt used. Textural data, however, showed some significant differences (p ≤ 0.01) between the breads and storage times. 5-hydroxymethylfurfural (HMF) is considered, nowadays, as an emerging ubiquitous processing contaminant; bread with the lowest level of Saltwell® had the lowest HMF content, and during storage, a decrease content was highlighted. Sensory data showed that the loaves had a similar rating (p ≤ 0.05) and differed only in salt content before storage. This study has found that durum wheat bread can make a nutritional claim of being "low in sodium" and "very low in sodium".

Effects of a Red Orange and Lemon Extract in Obese Diabetic Zucker Rats: Role of Nicotinamide Adenine Dinucleotide Phosphate Oxidase.

Diabetic nephropathy (DN) is the primary cause of end-stage renal disease, worldwide, and oxidative stress has been recognized as a key factor in the pathogenesis and progression of DN. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase has the most important contribution to reactive oxygen species generation during the development of DN. Bioactive compound use has emerged as a potential approach to reduce chronic renal failure. Therefore, a red orange and lemon extract (RLE) rich in anthocyanins was chosen in our study, to reduce the toxic renal effects during the development of DN in Zucker diabetic fatty rat (ZDF). RLE effects were examined daily for 24 weeks, through gavage, in ZDF rats treated with RLE (90 mg/kg). At the end of the experiment, ZDF rats treated with RLE showed a reduction of the diabetes-associated up-regulation of both NOX4 and the p47-phox and p22-phox subunits, and restored the BAX/BCL-2 ratio respect to ZDF rats. Furthermore, RLE was able to reduce the oxidative DNA damage measured in urine samples in ZDF rats. This study showed that RLE could prevent the renal damage induced by DN through its capacity to inhibit NOX4 and apoptosis mechanisms.

Change in taste-altering non-volatile components of blood and common orange fruit during cold storage.

Cold storage may cause changes in the volatile and non-volatile components of orange fruit, in association with the decrement of the characteristic fruit flavour and sensory acceptability. The aim of this work was to evaluate the changes of some non-volatile taste-altering components (total and individual sugars, acids, anthocyanins, putrescine and limonin) that may affect the organoleptic perception of cold-stored orange fruit. Three blood orange varieties ('Tarocco TDV', 'Tarocco Gallo', and 'Moro') and a common variety ('Washington navel') were stored at 6 ± 1 °C and 90-95% Relative Humidity (RH) for 60 d. Chemical and sensory assessments were performed during fruit storage at 15 d intervals. During storage, no dramatic change of the physicochemical parameters was recorded and the ascorbic acid content remained almost unchanged in all varieties. As expected, total anthocyanins significantly increased during storage. Limonin significantly decreased in all varieties. A consistent and significant increase in putrescine occurred during storage in the fruit of the pigmented varieties, not recorded in the common orange variety. Putrescine behaviour showed direct correlation with the accumulation of off-flavour in cold-stored 'Moro' and 'T. TDV' fruit, showing a clear influence of its relative concentration on the sensory perception of fruit. Finally, principal component analysis showed that the complete quality profile of the four investigated varieties represented clear differentiation without overlapping clusters. Our results suggest that the arise of a negative sensory perception in cold stored blood orange fruit might be linked to their accumulation of putrescine.

A red orange and lemon by-products extract rich in anthocyanins inhibits the progression of diabetic nephropathy.

The major cause of end-stage renal disease is the diabetic nephropathy. Oxidative stress contributes to the development of type II diabetes mellitus (T2DM). In this study we have evaluated the effect of a diet with a new standardized of red orange and lemon extract (RLE) rich in anthocyanins (ANT) in the progression of the kidney disease on Zucker diabetic fatty rats. Oxidative stress and renal function were analyzed. In diabetic rats, the RLE restored the blood glucose levels, body weight, and normalized the reactive oxygen species (ROS) total pathways. The kidney inflammation, in diabetic rats, has not shown significant change, showing that the oxidative stress rather than to inflammatory processes is a triggering factor in the renal complication associated with T2DM. Therefore, the administration of the RLE prevents this complication and this effect could be related to the inhibition of ROS production.

Bioactive compounds and antioxidant activity of four rose hip species from spontaneous Sicilian flora.

The morphological characteristics and health-promoting components (anthocyanins, phenolic compounds, carotenoids, and ascorbic acid) together with the in-vitro antioxidant capacity of hips collected from four rose species (Rosa canina, R. corymbifera, R. micrantha, and R. sempervirens) growing in Sicily (South Italy) were evaluated. The results revealed significant variability among the rose species as the hips collected from R. canina and R. sempervirens exhibited the highest total polyphenol contents (6784.5 and 6241.2 mg GAE/100 g DW, respectively) and highest antioxidant activities, whereas R. canina and R. micrantha hips had the highest total anthocyanin contents (2.94 and 3.86 mg CGE/100 g DW, respectively). The data showed that the analysed Sicilian rose hips have good nutritional quality, making them suitable as functional foods according to their biochemical composition and as promising sources of natural antioxidants. Also, these findings confirm that rose pseudo-fruits contain significant amounts of secondary metabolites that may positively affect human health.

Wholegrain Durum Wheat Bread Fortified With Citrus Fibers: Evaluation of Quality Parameters During Long Storage.

The aim of this work was to evaluate the effect of the addition of citrus fibers, from blood orange and lemon peels to produce a functional durum wheat bread. Breads fortified in fiber were packaged under a modified atmosphere (MAP) and stored at 25°C up to 120 days. No significant differences were observed with respect to the specific volume and weight, internal structure, pH and titratable acidity among the bread samples obtained using different types and percentages of fibers. Storage time, at 30 up to 90 days, affected significantly the bread firmness and caused significant differences in 5-hydroxymethylfurfural (HMF) levels in all bread samples. In fortified breads with citrus fibers the yeast and mold counts showed values of approximately 1 log10 cfu/g for the first 30 days and 3.5 log10 cfu/g at the end of storage. The results of the sensory evaluation highlight that loaves enriched with blood orange and lemon fibers showed a citrus flavor but had a similar overall evaluation respect to control bread produced without addition of citrus fiber. The results of this study showed that the addition up to 2% of blood orange and lemon fibers in wheat whole durum flour is a possible strategy to produce "high fibre" bread.

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 ß-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.

Evaluation of a Pomegranate Peel Extract as an Alternative Means 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.

Bioactive compounds in blood oranges (Citrus sinensis (L.) Osbeck): Level and intake.

Both the composition and the intake of antioxidants (anthocyanins, ascorbic acid and hydroxycinnamic acids) were reported for all blood oranges including the single cultivars (Moro, Tarocco and Sanguinello) and industrially produced juices. The mean values of the studied bioactive compounds in the edible part oranges were: 9.6mg/100g of orange edible part for the anthocyanins; 8.1, 0.7, 1.3, 3.8, 2.5mg/100g for total hydroxycinnamic acids, caffeic, sinapic, ferulic and coumaric acids, respectively and 59.1mg/100g for ascorbic acid. The consumption of blood oranges contributes to a daily intake of: 9.4mg/d (up to 55mg/d) of anthocyanins and 58.5mg/d (up to 340mg/d) of vitamin C, respectively. Data suggest that the 50% of consumers, males and females, receive more than the 70% and 90% of EAR value of vitamin C, respectively. The 25% of males and the 40% of females has an intake higher than the EAR.