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The purpose of this study is to analyze and characterize a Georgian red wine from Saperavi grape, obtained in Qvevri (Georgian traditional winemaking method), by using innovative techniques for the determination of the polyphenolic content, aroma, and its correlation to the sensory characteristics. This peculiar red wine, after high‐performance liquid chromatography with diode‐array detection and mass spectrometry (HPLC‐DAD‐MS), headspace solid‐phase microextraction–gas chromatography–mass spectrometry (HS‐SPME‐GC‐MS), and HS‐SPME‐GCxGC‐MS/TOF (two‐dimensional gas chromatography) chemical characterization showed a high polyphenol content (19.6 × 102 mg/L, 38.4% anthocyanins) and a wide range of volatile compounds, among which terpenes were associated with the aroma of flowers, lemongrass, and wood. Analyses were also conducted to determine the total polyphenol content correlated to antioxidant activity with the Folin–Ciocalteu spectrophotometric in vitro method (4.650 g GAE/L). In conclusion, for the first time on Saperavi wine, innovative techniques such as HPLC‐DAD‐MS, GC‐MS, and GCxGC‐MS/TOF were simultaneously applied in association with the traditional analytic techniques to perform a complete chemical characterization. These activities are part of a project about circular viticulture in the Georgian territory that will lead the production of traced quality wines and the valorization of the Georgian wine sector., BLURB FOR ETOC:Analysys and characterization of a Georgian red wine from Saperavi grape, obtained in Qvevri (Georgian traditional winemaking method), by using innovative techniques for the determination of the polyphenolic and key volatile compounds to produce traced quality wines and the valorisation of the Georgian wine sector.

BACKGROUND AND AIMS Saffron is the dried stigmas of Crocus sativus L. Crocus sativus L belongs to the family of Iridaceae and is mainly cultivated in several countries with mild and dry climates. It is one of the most expensive spices in the world and the main reason for its great cost is that saffron is still cultivated and harvested as it has been for millennia by hand. Saffron's name is derived from the Arab word for yellow, a name reflecting the high concentration of carotenoid pigments present in the saffron flowers’ stigmas which contribute most to the colour profile of this spice. Saffron contains numerous bioactive molecules known for their antioxidant activities, but also some such as picrocrocin, crocetin and Safranal (Sfr), Crocin (Crc) which are known to possess the ability to inhibit tumoral growth by inducing apoptosis. The aim of this study is to understand whether these four saffron-derived bioactive molecules display tumoral growth inhibition activity on kidney and bladder cancer cell lines. METHOD Saffron samples used for this purpose possess both different origins (Tuscany and Lombardy) and harvesting years (T_2017, T_2018, L_2017, L_2018) and, in addition, Sfr and Crc standard samples were also tested. Identification of Crocins, Safranal, Picrocrocin and Flavonoids was carried out by HPLC/DAD and HPLC/MS analysis. The identity of polyphenols was ascertained using data from HPLC/DAD analyses by comparison and combination of their retention times and UV/Vis spectra with those of authentic standards and previously reported data (Vignolini et al., 2008). The extract characterization is summarized in the table below. The extracts’ effects on cancer cell viability were evaluated through MTT assays and the tumoral cell lines chosen were Caki-1 and 786-O for the kidney and RT112 and RT4 for the bladder. MDCK cells were used as a healthy kidney model. RESULTS In the presence of both pure Sfr and Crc, post-hoc tests have shown a time and concentration-dependent decrease in viability for all the cancerous models in analysis, with more visible results observable for Crc, while having no effect on MDCK viability. Being Crc is the most represented molecule in all the extracts, before performing MTT assays, a normalization of this compound's concentration was carried out. L_2017, which contained the highest Sfr concentration among all the extracts tested, showed the highest inhibiting activity. Hypothesizing that Sfr was the molecule responsible for the decrease in cancer cell viability, all extracts and Sfr and Crc standards were tested on RT112 cells after normalizing the contents of these two molecules for each condition. We observed that Crc and Sfr alone and the Sfr-Crc mix had a small impact on cancer cell viability when compared with the effects of whole extracts. CONCLUSION Whole extracts demonstrated a higher impact on cancer cell viability compared with the standards tested both singularly and combined. This study, therefore, recognizes that numerous molecules in combination with Sfr and Crc have a role in reducing tumoral cell viability, demonstrating that the phyto-complex may possess higher therapeutic effects.

The by-products/wastes from agro-food and in particular the fruit industry represents from one side an issue since they cannot be disposed as such for their impact on the environment but they need to be treated as a waste. However, on the other side, they are a source of bioactive healthy useful compounds which can be recovered and be the starting material for other products in the view of sustainability and a circular economy addressing the global goal of zero waste in the environment. An updated view of the state of art of the research on fruit wastes is here given under this perspective. The topic is defined as follows: (i) literature quantitative analysis of fruit waste/by-products, with particular regards to linkage with health; (ii) an updated view of conventional and innovative extraction procedures; (iii) high-value added compounds obtained from fruit waste and associated biological properties; (iv) fruit wastes presence and relevance in updated databases. Nowadays, the investigation of the main components and related bioactivities of fruit wastes is being continuously explored throughout integrated and multidisciplinary approaches towards the exploitation of emerging fields of application which may allow to create economic, environmental, and social value in the design of an eco-friendly approach of the fruit wastes., info:eu-repo/semantics/publishedVersion

This is the accepted manuscript (post-peer reviewed) of the final paper: "In vitro studies on anti-inflammatory activities of kiwifruit peel extract in human THP-1 monocytes, published in Journal of Ethnopharmacology, Volume 233, 2019, Pages 41-46 https://doi.org/10.1016/j.jep.2018.12.044." -D'Eliseo D, Pannucci E, Bernini R, Campo M, Romani A, Santi L, Velotti F (2019): In vitro studies on anti-inflammatory activities of kiwifruit peel extract in human THP-1 monocytes. J Ethnopharmacol. 233:41-46. doi: 10.1016/j.jep.2018.12.044. -A B S T R A C T Ethnopharmacological relevance: Kiwifruit is native to eastern China and many are the references about the consumption of fruits and fruits extracts of the Actinidia plants in Chinese traditional medicine as therapeutic food supplements to prevent and/or counteract numerous disorders including inflammation-related diseases like cancer. Aim of the study: Aim of the present work was to obtain a kiwifruit peel extract, rich in polyphenols, and to explore the anti-inflammatory potential by analyzing its capability to target multiple pathways involved in monocyte-mediated inflammatory response. Materials and methods: The extract was obtained from the fruit peel of Actinidia deliciosa (A.Chev.) C.F.Liang & A.R.Ferguson, cv Hayward and characterized by HPLC-DAD-ESI-MS. Lipopolysaccharide-stimulated THP-1 monocytes were used as a model of human inflammation in vitro. Results: Analytical data evidenced that procyanidins resulted the main polyphenols present in the extract, representing the 92% w/w of the total. The extract inhibited the production of inflammatory molecules such as IL-6, IL-1β, TNF-α pro-inflammatory cytokines, HMGB1 danger signal and granzyme B serine protease by activated monocytes. In particular, an inhibitory activity of 81%, 68%, 63%, 76% and 60% on the extracellular release of IL-6, IL-1β, TNF-α, HMGB1 and granzyme B, respectively, was observed by western blot analysis. Moreover, the extract prevented STAT3 activation and promoted autophagy. Conclusions: The reported findings demonstrated a strong and broad anti-inflammatory profile of the kiwifruit peel extract, which makes it a promising preventive and therapeutic natural ingredient for nutraceutical, cosmetic and pharmaceutical formulations to counteract multiple inflammatory disorders.

The identification of natural bioactive compounds, able to counteract the abnormal increase of oxidative stress and inflammatory status in chronic degenerative non-communicable diseases is useful for the clinical management of these conditions. We tested an oral food supplement (OFS), chemically characterized and evaluated for in vitro and in vivo activity. Vitamin C, analyzed by High Performance Liquid Chromatography-Diode Array Detector (HPLC-DAD), was 0.19 mg/g in rosehip dry extract and 15.74 mg/capsule in the OFS. The identification of polyphenols was performed by HPLC-DAD, the total antioxidant capacity was assessed by Folin&ndash, Ciocalteu test. Total polyphenols were 14.73 mg/g gallic acid equivalents (GAE) for rosehip extract and 1.93 mg/g GAE for OFS. A total of 21 chronic kidney disease (CKD) patients and 10 healthy volunteers were recruited. The evaluation of routine laboratory and inflammatory parameters, erythrocyte glutathione transferase (e-GST), human oxidized serum albumin (HSAox), and assessment of body composition were performed at two different times, at baseline and after 5 weeks of OFS assumption. In the study, we highlighted a significant decrease of traditional inflammatory biomarkers (such as C-reactive protein, erythrocyte sedimentation rate, platelet to lymphocyte ratio) and other laboratory parameters like e-GST, azotaemia, and albuminuria after OFS treatment in CKD patients. Moreover, we demonstrated a lipid profile improvement in CKD patients after OFS supplementation.

Background and Aims Urinary tract infections (UTIs) are estimated to account for over 7 million office visits per year. UTIs are about 50 times more frequent in women than in men, however, in men the incidence steeply increases after 50 years of age for prostate involvement. UTI diagnosis is based on symptoms and urinalysis. Antibiotic therapy is often required in subjects with relapsing UTIs. Therefore, it would be useful to develop a therapeutic strategy based on natural active compounds, free from side effects (such as on renal function), that could counteract UTIs recurrence. For this purpose, tannins (TNs) could represent a valid therapeutic alternative. TNs are polyphenolic compounds known for their anti-inflammatory, antimicrobial and antioxidant effects. Recent studies highlighted how TNs intake can counteract UTI recurrence, improving the quality of life. The aim of our study was to evaluate the beneficial action of chestnut TNs administered as oral food supplement (OFS) in recurrent UTI patients. Method OFS was characterized by HPLC/DAD/MS analysis for its content in secondary polyphenolic metabolites and by Folin-Ciocalteau in vitro tests for its antioxidative and anti-free radical activity with stable radical DPPH•. For in vivo tests, 16 chronic kidney disease (CKD) patients (8 female and 8 males) who suffered from recurrent UTIs, were recruited. The OFS was administered in 2 cps/ day, for 9 weeks. Patients were evaluated at T0= baseline and T1= after 9 weeks of OFS treatment. We performed laboratory and urinary tests, and the quality of life (SCL-90R questionnaire) and body composition (bioimpedentiometry) assessment. All parameters and questionnaires were evaluated at T0 and T1. At the same time, oxidative stress (OS) and antioxidant defenses were monitored using CR4000 (FORD and FORT tests). Results HPLC/DAD/MS quali-quantitative analysis allowed for an in depth chemical characterization of the individual polyphenols present in the OFS: total tannins amount is 10.542mg/g, whereas total anthocyanins are 1.885mg/g. The Folin Ciocalteu assay results show a total polyphenols content of 68.7mg/g GAE; the antiradical activity by DPPH assay is 94% for a solution 0.519mg/mL of powder. The epidemiological features of the population were reported in Table 1 and it was divided into two subgroups, according to gender. Laboratory parameters were reported in Table 2. During the study, we observed a drop out of three female subjects because of gastrointestinal (GI) disorders (nausea, gastric heartburn). We did not find any significative change in body composition parameters after OFS treatment. We highlighted a significant reduction (Table 3) of ESR only in male patients (16,7± 2,2 mm/h vs 11,3± 1,5 mm/h, p=0,0062). The urinalysis, showed a significant reduction of leukocytes in both subgroups (male: 43,5 (1-450) n/uL vs 15 ± 5,7 n/uL p = 0,0391; female: 28,5 (1-990) n/uL vs 7 (1-91) n/uL p= 0,0625) but urinary bacterial flora, after OFS supplementation significantly decreased only in male subjects (428 ± 143,4 n/uL vs 34 (0- 450) n/uL p= 0,0156). Interestingly we observed a significant reduction of OS parameters after OFS treatment as reported in Table 4. We also observed a reduction of the anxious and depressive components in male subjects in the SCL-90R questionnaire. Conclusion TNs seem to exert an antimicrobial action in a gender dependent manner, useful to counteract the recurrence of UTIs. OFS contains Serenoa repens, a substance with peripheral, antispastic and antiandrogenic action that could induce nausea and vomiting. Female patients dropped out for GI disorders had lower body weight and SR probably were overdosed for their body mass. In male subjects, the decrease in anxiety and depression at the end of the study, could be linked to the improvement of the urinary symptomatology. A randomized clinical trial conducted a larger population is needed to confirm these findings and explain the correlation with gender.

Hydrolysable tannins, water-extracted from sweet chestnut (Castanea sativa Mill.) biomass (CHT) and membrane concentrated, have several remarkable effects as antioxidant, antimicrobial, and metal complexing agents. Raw CHT extract has been recognized as a plant biostimulant for its capacity to enhance plant rooting, and improve early P uptake (starter effect), as a seed-transplant seedling local treatment. It has been investigated, in particular on tobacco, also for its ability to enhance plant resistance to nematodes. More recently, some CHT fractions obtained by process stream fractioning and enriched in selected polyphenolic subclasses with antimicrobial and antioxidant effects were identified, chemically characterized and tested for their potential in promoting selected aspects of plant yield, quality and protection, and maintaining and improving feed and food quality during processing. A method of application of a CHT fraction with a biostimulant effect on plant root system was developed, to protect tobacco plant in Meloidogyne spp. infested fields. Results of this application consistently did not differ from better treatments.

The primary product of the oenological sector is wine. Nonetheless, the grape processing produces large amounts of by-products and wastes, e.g., the grape seeds. In the context of a sustainable production, there is a strong push towards reutilizing these by-products and waste for making useful derivatives since they are rich of bioactive substances with high additional value. As it is true for the wine itself, bringing these by-products derivatives to the market calls for quality measures and analytical tools to assess quality itself. One of the main objectives is to collect analytical data regarding bioactive compounds using potentially green techniques. In the present work, the profile of fatty acids and the main phenolic compounds were investigated by conventional methods. The qualitative analysis of the main functional groups was carried out by Fourier Transform Infrared (FTIR) spectroscopy. Moreover, the successful use of FTIR technique in combination with chemometric data analysis is shown to be a suitable analytical tool for discriminating the grape seeds. Grape seeds of different origin have different content of bioactive substances, making this technique useful when planning to recover a certain substance with specific potential application in health area as food supplement or nutraceutical. For example, Cesanese d’Affile seeds were found to have a rather high fat content with a significant fraction of unsaturated fatty acids. On the other hand, the seeds of Nero d’Avola exhibit the highest amount of phenolic compounds., This research received funding from Italcol SpA, Consulente Enologica Srl and the support of the Project NATUR-BAKERY-INNOV” Innovative production of a bakery line, for well-being and sport, based on functional natural extracts”—POR FESR 2014–2020—CUP 7429.31052017.113000254. Authors thank the support of the project: Nutraceutica come supporto nutrizionale nel paziente oncologico; CUP: B83D18000140007, info:eu-repo/semantics/publishedVersion

In the present study, the antifungal activities of two commercial tannins-rich dry fractions towards different filamentous fungi of agronomical and food interest were evaluated. In particular, a standardized fraction from sweet chestnut (Castanea sativa Mill.) wood by-products and a commercial green tea (Camellia sinensis L.) leaf extract were tested at different concentrations (0.1–5.0% and 0.2% w/v respectively). The Sweet Chestnut Wood fraction was produced in an industrial plant through an environmentally and economically sustainable process, involving hot-water extraction and a sequence of membrane filtration steps with different molecular cut-offs for fractionation and concentration of the active principles. The Sweet Chestnut Wood and Green Tea Leaf extracts were characterised via HPLC/DAD/MS quali-quantitative analysis. The first extract showed a polyphenolic content of 20.5% w/w, 100% hydrolysable tannins; the second one showed a polyphenolic content of 87.5% w/w, of which 96.2% epigallocatechin gallate and 3.8% epicatechin gallate. The antifungal activity of the Sweet Chestnut fraction in aqueous solutions was evaluated towards different filamentous fungi, in particular telluric phytopathogens (Fusarium oxysporum f. sp. radicis-lycopersici; Fusarium solani; Rhizoctonia solani; Sclerotium rolfsii) and post harvest pathogens (Botrytis cinerea, that can also attack field plants; Penicillium digitatum; Penicillium italicum), and compared to the activity of Green Tea Leaf extract solutions. The experimental results evidenced, for almost all tested fungi, inhibition of the mycelial growth rate in presence of tannins. The lowest inhibitions were observed for B. cinerea (7.5%, to 28.9%) and P. italicum (53.8% in 5.0% w/v Sweet Chestnut extract substrate). A proportional inhibitory effect to tannin concentration was observed for F. oxysporum f. sp. radicis-lycopersici and F. solani (from 33.7% to 56.6%), R. solani (from 29.7% to 68.8%) and P. digitatum (64.7% to 87.0%). The highest effect resulted for S. rolfsii, (5.0% to 100%).

Natural Product Research, DOI: 10.1080/14786419.2018.1508149 Antioxidant and anti-inflammatory effects of pomegranate peel extracts on bovine mammary epithelial cells BME-UV1 Fabio Mastrogiovanni a, Roberta Bernini a, Loredana Basiricò a,Umberto Bernabucci a , Margherita Campo b, Annalisa Romani b, Luca Santi a and Nicola Lacetera a aDepartment of Agricultural and Forestry Sciences (DAFNE), University of Tuscia, Viterbo, Italy; bDepartment of Statistics, Computing, Applications “Giuseppe Parenti” DiSIA, Phytolab, University of Florence, Florence, Italy Pomegranate peel extracts (PPE) were tested for the first time on BME-UV1, a valid cellular model to study the bovine mammary epithelial metabolism, to evaluate the effects on the oxidative stress and inflammatory status. Based on the statistical analysis of MTT data, PPE at 0.1, 1.0 and 10 lg/mL resulted not cytotoxic after 24 h, 48 h and 7 days of treatment. At the same concentrations, PPE induced a reduction of ROS production elicited by the addition of hydrogen peroxide or lipopolysaccharide evidencing an antioxidant effect confirmed also by a decrease of malondialdehyde. At 10 lg/mL, PPE reduced pro-inflammatory cytokines expressions showing an anti-inflammatory effect on BME-UV1 treated with lipopolysaccharide. Although in vivo experiments are necessary, the results of this study are promising for future applications of PPE as feed supplement for dairy cattle, in particular around calving, when the animals are more subject to oxidative stress and inflammatory diseases.

Food and agricultural waste represents a growing problem with negative effects on the economy, environment, and human health. Winemaking produces byproducts with high added value, which can be used for new productions in several application fields. From the perspective of biorefinery and circular economy, grape seeds could be exploited by extracting bioactive compounds with high added value before using biomass for energy purposes. The markets concerned are, in addition to the food, cosmetics, and pharmaceuticals sectors, which use bioactive compounds, the sector of biopolymeric materials and of energy for the production of biohydrogen and biomethane. Generally, bioactive components should be investigated through an integrated and multidisciplinary study approach based on emerging analytical techniques; in this context, attention is addressed towards green and sustainable procedures; an update of extraction techniques, innovative technologies, and chemometrics are described. Nowadays, processes so far tested on a pilot scale for grape waste are developed to enhance the extraction yields. Here, a picture of the Italian experience applied to the byproducts of the wine industry is given.

Sweet Chestnut ( Castanea sativa Mill.) wood extracts, rich in Hydrolyzable Tannins (HTs), are traditionally used in the tanning and textile industries, but recent studies suggest additional uses. The aim of this work is the HPLC-DAD-ESI-MS characterization of Sweet Chestnut aqueous extracts and fractions obtained through a membrane separation technology system without using other solvents, and the evaluation of their antioxidant and antiradical activities. Total tannins range between 2.7 and 138.4 mM; gallic acid ranges between 6% and 100%; castalagin and vescalagin range between 0% and 40%. Gallic Acid Equivalents, measured with the Folin-Ciocalteu test, range between 0.067 and 56.99g/100g extract weight; ORAC test results for the marketed fractions are 450.4 and 3050 μmol/g Trolox Equivalents/extract weight. EC50 values, measured with the DPPH test, range between 0.444 and 2.399 μM. These results suggest a new ecofriendly and economically sustainable method for obtaining chestnut fractions with differentiated, stable and reproducible chemical compositions. Such fractions can be marketed for innovative uses in several sectors.

Protection of plants against bacterial diseases still mainly relies on the use of chemical pesticides, which in Europe correspond essentially to copper-based compounds. However, recently plant diseases control is oriented towards a rational use of molecules and extracts, generally with natural origin, with lower intrinsic toxicity and a reduced negative environmental impact. In this work, polyphenolic extracts from vegetable no food/feed residues of typical Mediterranean crops, as Olea europaea, Cynara scolymus, and Vitis vinifera were obtained and their inhibitory activity on the Type Three Secretion System (TTSS) and the Quorum Sensing (QS) of the Gram-negative phytopathogenic bacterium Pseudomonas savastanoi pv. nerii strain Psn23 was assessed. Extract from green tea (Camellia sinensis) was used as a positive control. Collectively, the data obtained through gfp-promoter fusion system and real-time PCR show that all the polyphenolic extracts here studied have a high inhibitory activity on both the TTSS and QS of Psn23, without any depressing effect on bacterial viability. Extracts from green tea and grape seeds were shown to be the most active. Such activity was confirmed in planta by a strong reduction in the ability of Psn23 to develop hyperplastic galls on explants from adult oleander plants, as well as to elicit hypersensitive response on tobacco. By using a newly developed Congo red assay and an ELISA test, we demonstrated that the TTSS-targeted activity of these polyphenolic extracts also affects the TTSS pilus assembly. In consideration of the potential application of polyphenolic extracts in plant protection, the absence of any toxicity of these polyphenolic compounds was also assessed. A widely and evolutionary conserved molecular target such as Ca2+-ATPase, essential for the survival of any living organism, was used for the toxicity assessment.