Protective Effects of a Red Orange and Lemon Extract (RLE) on the Hepatotoxicity Induced by Ochratoxin A in Rats.

Ochratoxin A (OTA) is a highly potent mycotoxin that contaminates many kinds of food and feed sources. Its significant impact on human health and animal productivity makes it a topic of particular concern. The role of specific bioactive compounds used as dietary antioxidants is believed to be substantial due to their capacity to act as free radical scavengers. Because of the well-known oxidative stress induced by OTA, the primary objective of this work was to evaluate the antioxidant effects of a standardized powder extract recovered from citrus processing waste, red orange and lemon extract (RLE), on liver damage induced by OTA in a rat model. This study aimed to examine the impact of oral administration of RLE (90 mg/kg b.w.) on hepatic function and oxidative balance in Sprague-Dawley rats (n = 6/group) treated with OTA (0.5 mg/kg b.w.) over a period of 14 days. The administration of OTA alone resulted in both biochemical changes and an imbalance in redox status in the liver. However, the use of RLE alleviated the activity of antioxidant enzymes and dramatically decreased the serum levels of ALT (alanine aminotransferase), AST (aspartate aminotransferase), and ALP (alkaline phosphatase), providing evidence of its protective benefits. Based on the findings from liver histology tests, the administration of RLE resulted in mitigation of lymphoplasmacytic inflammation, steatosis, and necrosis in the OTA group. These results indicate that the novel phytoextract RLE holds potential for application in the field of nutraceuticals.

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.

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.

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.

Productive Performance and Meat Characteristics of Kids Fed a Red Orange and Lemon Extract.

This study evaluates the animal performance and meat characteristics of 60 Saanen suckling kids daily fed a red orange and lemon extract (RLE), rich in anthocyanins. In our methodology, after colostrum administration, animals are randomly assigned to two treatments: Treatment group (Group RLE; n = 30) that received RLE (90 mg/kg live body weight) as oral food additive, and a control group (Group CON; n = 30) that received a standard diet. Animals are slaughtered after 40 days. The RLE administration did not influence daily weight gain, carcass measurements, or incidences (expressed as a percentage) of different anatomical regions on the whole carcass weight. On the contrary, RLE supplementation significantly improved the oxidative profile of the meat seven days after slaughtering, as demonstrated by the reduced levels of thiobarbituric acid reactive substances (TBARS; p < 0.01) and hydroperoxides (p < 0.01) in Group RLE compared to Group CON. A significant influence of RLE administration is observed on day 7 for yellowness (p < 0.01). There are also lower saturated and higher monounsaturated and polyunsaturated fatty acids concentration in Group RLE meat (p < 0.01), which also shows lower atherogenic and thrombogenic indexes (p < 0.01) compared to Group CON. The study demonstrates that the supplementation of a diet with RLE rich in anthocyanins is effective to improve the meat quality.

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.

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.

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 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.

Beneficial Effects of Pomegranate Peel Extract and Probiotics on Pre-adipocyte Differentiation.

The beneficial effects of pomegranate are due to the ellagitannins and anthocyanins content, which are protective toward a wide variety of diseases including inflammatory diseases. Many investigators have reported that pomegranate waste (peel and seeds) extracts, made from waste product of industrial processing, show free radical scavenger and a potent antioxidant capacity. Pomegranate extracts (PEs) were also reported to possess noteworty antibacterial, antiviral, hypolipidemic, and anti-inflammatory bioactivities thanks to the polyphenolic compounds content, which includes punicalagins, gallic acid, and ellagic acid derivatives. The focus of the present manuscript was to study the prebiotic potentiality of a PE, soluble in water, and characterized through HPLC-PDA-ESI/MS n for its phenolic content. Moreover, since it has been reported that pomegranate extracts decreased the level of lipids in the blood and that a number of probiotic strains have been shown to affect adipogenesis in cell culture, this study was also performed to test the in vitro effects of PE and probiotic L. rhamnosus GG ATCC 53103 strain (LGG) on 3T3-L1 cell line. PE and probiotics substantially reduced the triglyceride content and intracellular lipid increase, compared to the control group. However, the combination treatment of PE and LGG filtered spent broth (SB) was the most effective in reducing triglyceride content and intracellular lipid accumulation. The mRNA expression levels of the main transcriptional factors implicated in adipocyte differentiation were substantially lower in 3T3-L1 cells treated with PE and LGG filtered SB. These results evidenced that a synergistic effect of probiotics and polyphenols contained in PE may affect in vitro adipogenesis and may contribute in development of new nutraceutical/probiotic-based remedies to prevent and to treat obesity.

Transcriptomic Analysis of Orange Fruit Treated with Pomegranate Peel Extract (PGE).

A Pomegranate Peel Extract (PGE) has been proposed as a natural antifungal substance with a wide range of activity against plant diseases. Previous studies showed that the extract has a direct antimicrobial activity and can elicit resistance responses in plant host tissues. In the present study, the transcriptomic response of orange fruit toward PGE treatments was evaluated. RNA-seq analyses, conducted on wounded fruits 0, 6, and 24 h after PGE applications, showed a significantly different transcriptome in treated oranges as compared to control samples. The majority (273) of the deferentially expressed genes (DEGs) were highly up-regulated compared to only 8 genes that were down-regulated. Gene Ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis showed the involvement of 1233 gene ontology (GO) terms and 35 KEGG metabolic pathways. Among these, important defense pathways were induced and antibiotic biosynthesis was the most enriched one. These findings may explain the underlying preventive and curative activity of PGE against plant diseases.

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.

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.

Anthocyanins in different Citrus species: an UHPLC-PDA-ESI/MSn -assisted qualitative and quantitative investigation.

BACKGROUND: Anthocyanins are water-soluble pigments belonging to the flavonoid family. They are typically present in the flesh and peel in the blood orange cultivars. Although blood orange young shoots and flowers are not anthocyanin-colored, lemon, citron, rangpur lime, and Meyer lemon young shoots and flowers exhibit marked pigmentation as a result of anthocyanins, demonstrating that anthocyanin biosynthesis in the Citrus genus is both tissue- and genotype-dependent. The present study aimed to examine the qualitative and quantitative anthocyanin profile of fruit and other tissues from different Citrus species. RESULTS: The presence of anthocyanin-pigmented stigmas in the young flowers of a blood orange tree (cv. 'Moro') was characterized and reported for the first time. The dominant pigments in blood orange fruits were cyanidin 3-glucoside and cyanidin 3-(6''-malonyl-glucoside), whereas different patterns were observed in the young shoots, flowers and peel tissues of different Citrus species. CONCLUSION: The present study is the first to report differentially expressed anthocyanin pigmentation patterns in different organs from several species of the genus Citrus. The results obtained could also represent a starting point for further investigations that aim to better understand which regulatory genes are involved in anthocyanin biosynthesis in the fruits, shoots and floral tissues of different Citrus species. © 2016 Society of Chemical Industry.

Screening of the anthocyanin profile and in vitro pancreatic lipase inhibition by anthocyanin-containing extracts of fruits, vegetables, legumes and cereals.

BACKGROUND: The phytotherapic treatment of overweight and/or moderate obesity is growing widely, thus there is a great interest towards the phenolic compounds of fruits and vegetables which may inhibit pancreatic lipase enzyme. In this study, we report the chemical composition and in vitro pancreatic lipase inhibitory activity of 13 freeze-dried anthocyanin-containing extracts of different Mediterranean plants: fruits (blood orange, pomegranate, blackberry, mulberry and sumac), citrus by-products (blood orange peel), citrus vegetative tissues (young lemon shoots); vegetables (red cabbage and violet cauliflower), legume seeds (black bean), cereals (black rice), and cereal processing by-products (black rice hull). Total phenols and anthocyanins were determined. Individual anthocyanins were identified by UHPLC-PDA-ESI/MSn . RESULTS: Results revealed a wide variation in the distribution of anthocyanin compounds. Blood orange and pomegranate juice extracts had the highest total anthocyanin content and exhibited the strongest inhibition of pancreatic lipase in vitro. CONCLUSION: Inhibitory activity was positively correlated with anthocyanin content. In appropriate formulations, anthocyanin-containing extracts could find a use as anti-obesity agents. © 2016 Society of Chemical Industry.

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.