Corrigendum: Correlation between antioxidant and anticancer activity and phenolic profile of new Apulian table grape genotypes (V. Vinifera L.).

[This corrects the article DOI: 10.3389/fpls.2022.1064023.].

Beneficial Effects of Table Grape Use on Serum Levels of Omega-3 Index and Liver Function: A Randomized Controlled Clinical Trial.

This clinical trial was aimed to investigate the effects of fresh table grape intake on the serum levels of the Omega-3 index, defined as the sum of eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) levels. Forty consecutive healthy subjects were randomly assigned to the control group, receiving only dietary recommendations, and the grape group receiving a daily dose of 5 g of fresh table grape per kg of body weight, for 21 days. Compared with baseline, the grape treatment produced no significant difference in the serum levels of glucose, liver transaminase, and triglycerides, with the exception of cholesterol value, which was significantly reduced in both control and grape group (180.5 ± 20.32 vs. 196.1 ± 30.0 and 181.4 ± 21.9 vs. 194.3 ± 37.5, respectively). After 4 weeks from the end of grape treatment, the analysis of single fatty acids showed a significant increase in oleic acid content (14.15 ± 1.8 vs. 12.85 ± 1.6, p < 0.05) and a significant induction of the Omega-3 index (8.23 ± 1.9 vs. 6.09 ± 1.2, p < 0.05), associated with increased serum levels of adiponectin (24.09 ± 1.08 vs. 8.8 ± 0.7, p < 0.001). In contrast, the expression of fibroblast growth factor 21 (FGF21), a molecule associated with metabolic syndrome and liver disease, was significantly reduced (37.9 ± 6.8 vs. 107.8 ± 10.1, p < 0.001). The data suggest that the intake of fresh grape improves the Omega-3 index in the serum and exerts beneficial effects on liver function through the overexpression of adiponectin and the reduction in FGF21 levels.

Nutraceuticals: Focus on Anti-Inflammatory, Anti-Cancer, Antioxidant Properties in Gastrointestinal Tract.

In recent years, nutraceuticals have gained great popularity, owing to their physiological and potential health effects, such as anti-inflammatory, anti-cancer, antioxidant, and prebiotic effects, and their regulation of lipid metabolism. Since the Mediterranean diet is a nutritionally recommended dietary pattern including high-level consumption of nutraceuticals, this review aimed to summarize the main results obtained by our in vitro and in vivo studies on the effects of the major constituents of the Mediterranean diet (i.e., extra virgin olive oil compounds, polyunsaturated fatty acids, and fruit components). Based on experimental studies, the therapeutic purpose of nutraceuticals depends on their bioavailability, solubility, toxicity, and delivery system. This review provides more in-depth knowledge on the effects linked to nutraceuticals administration on human health, focusing the gastrointestinal tract and suggesting specific dietary components for personalized adjuvant therapies.

Correlation between antioxidant and anticancer activity and phenolic profile of new Apulian table grape genotypes (V. Vinifera L.).

Grapes represent a significant source of phenolic compounds known for their health-promoting properties, such as antioxidant capacity on normal cells and prooxidant activity on tumor cells. The genotype highly affects the polyphenolic composition in grapes and, consequently, the nutritional quality of berries. This work aimed to characterize the phenolic composition, the antioxidant, and anticancer activity of grape skin extracts (GSEs) of nine new table grape genotypes selected from a breeding program to obtain new cultivars of seedless table grapes, well adapted to the climatic change and with higher nutraceutical properties. The grape polyphenolic profile was characterized by Ultra-High-Performance Liquid Chromatography/Quadrupole-Time of Flight mass spectrometry analysis. GSE antioxidant activity was determined by the ABTS, DPPH, and ORAC assays; GSE cell growth inhibition test was carried out in the Caco2 human cancer cell line. The nine GSEs showed different flavonoid and non-flavonoid profiles, and all possessed antioxidant activity, with the 'Aika N.', 'Turese N.', and 'Egnatia N.' the most active. As anticancer activity against the tested cancer cell line, 'Daunia N.' and 'Apenestae N.' showed the EC50 after 24 h of 35.60 µg/mL and 150.91 µg/mL, respectively. The relationship between polyphenolic profile and the antioxidant and anticancer activity of GSE was also investigated. Interestingly, among the different classes of polyphenolics, flavan-3-ols e proanthocyanidins showed the highest positive correlation with the anticancer activity of extracts. These findings can be helpful for the preparation of new extracts for the pharmaceutical and nutraceutical industry and geneticists working in vine breeding programs.

Impact of Fresh Table Grape Intake on Circulating microRNAs Levels in Healthy Subjects: A Significant Modulation of Gastrointestinal Cancer-Related Pathways.

SCOPE: The study aims to investigate the effects of fresh table grape consumption in healthy subjects on circulating levels of the most common human microRNAs (miRNAs). The regulatory network governed by these modulated miRNAs is also investigated. METHODS AND RESULTS: Autumn Royal table grape, used in this study, is chosen for its high polyphenolic content and antioxidant properties. The study is a randomized controlled trial, in which 40 consecutive subjects are recruited on a voluntary basis and randomly assigned to two groups of the study, the control group, receiving only dietary recommendations and a grape group receiving a daily dose of 5 g of fresh table grape per kg of body weight for 21 days. All analyses are performed at baseline and after 21 days of dietary treatment. Circulating miRNAs levels are detected by Real-Time quantitative PCR (RT-qPCR) followed by bioinformatic functional analysis. The study identifies 20 circulating miRNAs differentially expressed in healthy subjects after grape intake, and in particular, 18 of 20 are down-regulated and 2 are up-regulated. CONCLUSION: The dietary intake of table grape affects circulating miRNAs levels in healthy subjects, particularly the miRNAs related to pathways involved in counteracting cancer development, including gastrointestinal cancers.

Classification of wine grape biotypes according to their variety and sanitary condition by fingerprinting untargeted analysis.

Grapes secondary metabolites content mainly depends on variety, but also on climate and cultural conditions, including sanitary status. This study aimed to use a metabolomic fingerprinting approach for grouping 72 wine grape biotypes, Negro amaro n. (N), Malvasia nera di Brindisi/Lecce n. (M), and Uva di Troia n. (U), on the basis of their cultivar and virological conditions. The skins were extracted and analysed by flow injection mass spectrometry; a one-way ANOVA/Principal Component Analysis (PCA) allowed to efficiently cluster the samples, recognizing M from N and U biotypes. Conversely, the clusterisation of the biotypes affected by different virus complexes was really more tough and a clear distinction among infected plants was not always observed. However, very interestingly, by applying ANOVA/PCA to the biotypes of each varieties, singularly, healthy biotypes were sharply separated in all the varieties and a relationship between anthocyanin compounds and Grapevine leafroll associated virus (GLRaV3) slightly appeared.

Microarray data and pathway analyses of peripheral blood mononuclear cells from healthy subjects after a three weeks grape-rich diet.

Using Human Gene Expression Microarrays (Agilent) technologies, we investigated changes of the level of gene expression in peripheral blood mononuclear cells of healthy subjects after 21 days of fresh table grape-rich diet and after an additional 28-day washout. Several hundreds of genes were differentially expressed after grape intake or after washout. The functional analysis of these genes detected significant changes in key processes such as inflammation and immunity, thrombosis, DNA and protein repair, autophagy and mitochondrial biogenesis. Moreover, fresh grape intake was found to influence the expression of many long non-coding RNA genes. The data can be valuable for researchers interested in nutrigenetics and nutrigenomics studies and are related to the research article "Gene expression signature induced by grape intake in healthy subjects reveals wide-spread beneficial effects on PBMCs" [1].

Optimized ultrasonic-assisted extraction of phenolic antioxidants from grape (Vitis vinifera L.) skin using response surface methodology.

Currently, the analysis of bioactive compounds is easily carried out by modern chromatographic and spectrometric techniques, but the success of the procedure still depends on the efficiency of the extraction methods. This work aimed at finding the optimal conditions for ultrasonic-assisted extraction (UAE) of phenolic antioxidants from grape skin using single-factor experimentation combined with response-surface methodology. Our extraction parameters were ethanol concentration, time and temperature. Total phenolic content (TPC and TPC-280), 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity and oxygen radical absorbance capacity (ORAC) were dependent variables. A second-order polynomial model satisfactorily described the experimental data. Optimized extraction conditions for UAE from grape skin were: 59.5% ethanol, 113.6 min and 66.8 °C for TPC extraction; 61.1% ethanol, 99.4 min and 66.1 °C for TPC-280 extraction; 57.1% ethanol, 99.5 min and 66.8 °C for DPPH radical-scavenging activity; 60.2% ethanol, 82.4 min and 66.8 °C for ORAC value. Our work provides detailed and useful information to optimize the extraction of grape skin phenols, which are widely used as dietary supplements and nutraceuticals.

Ampelometric Leaf Trait and SSR Loci Selection for a Multivariate Statistical Approach in Vitis vinifera L. Biodiversity Management.

High estimated heritability values were recently revealed for mature leaf traits in grape (Vitis vinifera L.), thus redeeming ampelography in the era of molecular markers. The "Organisation Internationale de la Vigne et du Vin (OIV)" set a list of hundreds of descriptors for grapevine in order to standardize ampelographic and ampelometric scores. Therefore, the selection and reduction of the number of OIV codes can represent a major goal for leaner biodiversity assessment studies. The identification of ampelometric traits associated with grape diversity allows to construct Classification Trees with chi squared automatic interaction detection (CHAID) algorithm, a stepwise model-fitting method that produces a tree diagram in which at each step the sample pool is splitted based on the independent variables statistically different for the dependent variable. A collection of 100 table and wine grapevines (Vitis vinifera L.) was characterized and evaluated by means of six microsatellites and twenty-two ampelometric traits on mature leaves. Nine ampelometric traits were selected by principal component analysis and employed to build the classification trees based on CHAID algorithm. The strategy can represent an effective tool for grape biodiversity management, right allocations, and identification of new grape genotypes, implemented by a further microsatellite investigation only when unsolved cases occur, allowing faster and cheaper results.

Morphological variability in leaves and molecular characterization of novel table grape candidate cultivars (Vitis vinifera L.).

The present work report the characterization of twenty-one table grapes candidate cultivars plus five registered ones included as reference, by means of 47 ampelographic traits, 23 ampelometric measurements and six microsatellite loci. The final goal of the research was to analyse the possibility of reducing the number of morphological and molecular tools required for a precise and effective description of a grape genotype or cultivar. This would be of great help for future biodiversity description on a larger sample of more than 300 table grapes accessions today grown at the 'Consiglio per la Ricerca e la sperimentazione in Agricoltura (C.R.A.)-Unità di ricerca per l'uva da tavola e la vitivinicoltura in ambiente mediterraneo (Bari-Italy)'. OIV ampelographic traits showed a clear distinction among all twenty-six genotypes analysed, suggesting the relevant morphological variability investigated. Principal component analysis based on ampelometric traits revealed main veins ON(3), ON(4) and O(3)N(4); ratios between main veins; angles between main veins and of petiolar sinus, to be the most effective records in differentiating cultivars, for a total variation of 69.9 % described by the first three components. Molecular analysis based on six microsatellite loci was performed on all genotypes, providing a detailed molecular profile and a dendrogram of genetic similarity, in which all genotypes were clearly distinguishable. Finally, with the goal of using the minimum possible number of markers to differentiate genotypes, microsatellites VVMD5 and VVMD27 were selected to be sufficient to distinguish among all the candidate cultivars included in the analysis, representing a possible 'step by step' approach when a molecular characterization has to be undertaken on a large number of genotypes, by first testing few markers and increasing their number only if necessary.

Sangiovese and its offspring in southern Italy.

This paper demonstrates the importance of different approaches such as ampelography, historical researches, and molecular analysis to reveal direct parent-child relationship. The aim of this paper was to highlight the degree of relationship to five varieties spread in southern Italy, through ampelographic and molecular characterization: Sangiovese, Mantonico di Bianco, Gaglioppo di Cirò, Mantonicone, and Nerello Mascalese. Molecular characterization was carried out through 52 SSR molecular markers, showing that Sangiovese and Mantonico di Bianco are the parents of Gaglioppo di Cirò, Mantonicone, and Nerello Mascalese. Ampelographic description was performed using the method developed by the Organisation Internationale de la Vigne et du Vin. This analysis identifies three distinct groups: the first brings together Sangiovese and the two offspring Nerello Mascalese and Gaglioppo di Cirò, while Mantonico di Bianco and Mantonicone are positioned at a distance from the first and between them. Using molecular characterization, supported by the ampelographic one, we showed that Gaglioppo di Cirò, Mantonicone, and Nerello Mascalese, three varieties recovered in the southern regions of Italy, such as Calabria and Sicily, originated by the cross between a nationally spread grape variety as Sangiovese and a Calabria autochthonous vine as Mantonico di Bianco.

Evidences for an alternative genealogy of 'Sangiovese'.

Two different hypothesis for the parentage of 'Sangiovese', the most important and widespread Italian winegrape, have been proposed by some previous studies. We screened our grapevine collection, mostly comprising south Italian cultivars collected to preserve biodiversity, to asses kinships. Surprisingly we found two previously unreported candidate parents for 'Sangiovese'. The first putative parent is 'Ciliegiolo' a well know variety already addressed as relative of 'Sangiovese'; the second putative parent is 'Negrodolce', an old local variety we recovered and was considered lost during the last century. In order to obtain a stronger statistical support for this new kinship, we tested seventy different microsatellite markers but only 57 were found reliable. The new proposed parentage stood well even with such a in depth molecular analysis whereas only one discrepancy was found in one of the 57 microsatellite marker analyzed. This discrepancy is certainly due to a null-allele and therefore it should not impair our hypothesis but it points out limits of the microsatellites profiling as a pedigree research method considering that this is the third different kinship proposed so far for 'Sangiovese'. Thus in this article, by means of detailed molecular fingerprinting, we provide a completely new strong evidence for a south Italian origin of 'Sangiovese' and we discuss our findings comparing our data with those previously reported by other authors.

HPLC-DAD-ESI-MS analysis of flavonoid compounds in 5 seedless table grapes grown in Apulian Region.

UNLABELLED: Flavonoids present in skin extracts of red seedless table grape varieties Summer Royal, Autumn Royal, and Crimson, and white seedless varieties Carati and Thompson were analyzed by HPLC-DAD-MS, in 3 y of study (2006 to 2008). The anthocyanins, delphinidin-3-O-glucoside, cyanidin-3-O-glucoside, petunidin-3-O-glucoside (with their corresponding p-coumaroyl derivatives), peonidin-3-O-glucoside, and malvidin-3-O-glucoside (with their corresponding acetyl, caffeoyl, and p-coumaroyl derivatives) were found. In addition the flavonols quercetin-3-O-glucuronide, quercetin-3-O-rutinoside, quercetin-3-O-glucoside, kaempferol-3-O-glucoside, kaempferol-3-O-galactoside, and the flavan-3-ols procyanidin B1, procyanidin B2, and catechin were also detected. Anthocyanins were the main flavonoids in red grapes ranging from 24 (Crimson) to 500 (Summer Royal) mg/kg fresh weight of grapes; consistent levels of flavonols and flavan-3-ols were also quantified in all varieties. To determine the effective climatic influence on flavonoids content in field conditions, viticultural practices have been developed, that could exclude the effects of direct solar radiation from confounding the assessment of those related to thermal conditions alone. A strong positive correlation was determined between flavonoids and temperature data that seem to be responsible for the difference of these metabolites along the years; furthermore, it has been possible to define a linear relationship (R(2) = 0.6871, P = 0.0057) between thermal amplitude and total flavonoids values in the red grapes. PRACTICAL APPLICATION: Grapes are economically the most important fruit species in the world and approximately 30% of its production is used as fresh fruit. Because of the very important role of flavonoids in food quality as well as their health-promoting properties, and considering that our experiments were performed along 3 consecutive years, gathered results in this research are quite promising to give a useful information on the flavonoid contents and their evolution in 5 seedless table grapes that are widespread in Mediterranean regions but also in California and South America, and are grown in a viticultural climate (Apulia, South Italy) very close to some regions of Spain, Turkey, Tunisia, and Israel.

Analysis of high-identity segmental duplications in the grapevine genome.

BACKGROUND: Segmental duplications (SDs) are blocks of genomic sequence of 1-200 kb that map to different loci in a genome and share a sequence identity > 90%. SDs show at the sequence level the same characteristics as other regions of the human genome: they contain both high-copy repeats and gene sequences. SDs play an important role in genome plasticity by creating new genes and modeling genome structure. Although data is plentiful for mammals, not much was known about the representation of SDs in plant genomes. In this regard, we performed a genome-wide analysis of high-identity SDs on the sequenced grapevine (Vitis vinifera) genome (PN40024). RESULTS: We demonstrate that recent SDs (> 94% identity and >= 10 kb in size) are a relevant component of the grapevine genome (85 Mb, 17% of the genome sequence). We detected mitochondrial and plastid DNA and genes (10% of gene annotation) in segmentally duplicated regions of the nuclear genome. In particular, the nine highest copy number genes have a copy in either or both organelle genomes. Further we showed that several duplicated genes take part in the biosynthesis of compounds involved in plant response to environmental stress. CONCLUSIONS: These data show the great influence of SDs and organelle DNA transfers in modeling the Vitis vinifera nuclear DNA structure as well as the impact of SDs in contributing to the adaptive capacity of grapevine and the nutritional content of grape products through genome variation. This study represents a step forward in the full characterization of duplicated genes important for grapevine cultural needs and human health.