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the problem of party regulation according to the Palermo school of public law. between past and future. In particular, the analysis is on three authors: Pietro Virga, Guido Corso and Marco Mazzamuto.

The present study investigated the effects of different grape pomace storage techniques on the effectiveness as feed on in vitro ruminant digestion efficiency. Grape pomace from an autochthonous red grape variety (cv Nero di Troia) was used as fresh (GP) or ensiled, both without additives (SIL) and with the addition of a bacterial strain, Lactiplantibacillus plantarum 5BG (SIL+). All the different storage treatments were subject to chemical and microbiological evaluation, as well as in vitro digestibility, and gas production. Microbiological data revealed the good quality of grape pomace and silages due to the lactic acid bacteria populations and low presence, or absence, of undesirable microorganisms. The addition of L. plantarum 5BG influenced the chemical characteristics of the silage (SIL+). Ensiling technique deeply changed the polyphenolic composition, reducing anthocyanins, flavonols, and flavanols (condensed tannins precursors), particularly when L. plantarum 5BG was added. Antioxidant capacity was reduced by ensiling, in correlation with the polyphenolic content decrease. The oxygen radical absorbance capacity (ORAC) value of SIL+ was the lowest (P < 0.01) and its total phenol content was lower than SIL (P < 0.01). No statistical differences were observed between GP, SIL, and SIL+ on the antioxidant capacity by TEAC assay (P > 0.05). Ensiling did not affect the grape pomace nutrient profile, except for the reduction in NFC content. Apparent in vitro digestibility showed how ensiling increased dry matter (DM), organic matter (OM), neutral detergent fiber (NDF), crude protein (CP), ether extract (EE), and non-fiber carbohydrates (NFC) disappearance (P < 0.01), particularly with the L. plantarum 5BG inoculation. Moreover, SIL+ showed the lowest propionic acid (P < 0.05) and the highest methane (P < 0.01), butyric acid (P < 0.01), and nitrogen (P < 0.05) in vitro production. Ensiling GP resulted in a better in vitro digestibility, particularly if L. plantarum 5BG strain is added, probably due to the reduction of flavanols and their lower microbial activity inhibition.

What makes Guarino’s figure so fascinating is the multiplicity of his interests, combined with the long activity of his life as a jurist, which ensure that there is not, just one Guarino, but as many as there are seasons and events that he has found to investigate with the strength of reason, clarity, objectivity, combined with a rigorous method of investigation that dissolves into an inexhaustible constructive fantasy. His thought is to separate time between a before and an after, producing a clear discontinuity with the past and creating new perspectives for the future.This essay wants to investigate two central points in Guarino's reflection: the costituzione materiale and the task of jurists. His speech starts, in a certain sense, when everything is now to be built: that is, in the aftermath of the gigantic tragedy of the Second World War. The "postwar" has confronted scholars of public law with the reality of a structure which, however, is always the expression of certain ideas and certain interests, as they are asserted by certain political forces. Guarino starts from this point, from the awareness that every community gives life to an order that is expressed in a system of rules and at the same time in a system of relationships, but that we must not stop at formal data alone. In Guarino's thought, the way that solidly roots the figure of the jurist in human society and history lies in the ability to stretch out on reality and consider oneself a participant in the construction of the legal system. It is up to subsequent generations of jurists to collect not only a cultural legacy of extraordinary richness, but also, and I would say more than anything else, the example of a jurist informed by the sense of trust in institutions and in man

Valorization of the neglected Prunus mahaleb L., fruits as a source of functional molecules Gerardi C.1, Frassinetti S.3, Leone A.1, Calabriso N.2, Carluccio M.A.2, F. Blando1 and Mita G. 1 1 Institute of Sciences of Food Production, CNR, Lecce, Italy 2 Institute of Clinical Physiology, CNR, Lecce, Italy 3 Institute of Biology and Agricultural Biotechnology, CNR, Pisa, Italy Abstract: Prunus mahaleb L. is a tree producing dark-red small stone fruits, not used for fresh consumption due to their astringent and sour taste. In this communication will be described a "mahaleb fruit concentrated extract" (mfce) and its chemical characterization. The extract showed high anthocyanin, flavonol and coumarin content. Moreover, the results of the in vitro assays for mfce biological activities will be reported. Mfce showed strong antioxidant capacity and anti-proliferative, pro-apoptotic, anti-inflammatory and anti-mutagenic effects. This study demonstrated several healthful effects of mfce that may be of interest as natural food colorant and as source of molecules for the formulation of functional foods.

Introduzione Prunus mahaleb L. è un arbusto deciduo che cresce in boschi radi anche su terreni marginali e in climi aridi. L'albero è nativo dei paesi del mediterraneo e dell'Asia centrale e produce dei piccoli frutti simili a ciliegie di un colore quasi nero, non commestibili a causa del loro sapore amaro. Studi precedenti hanno dimostrato un elevato contenuto in antocianine in questi frutti, che suggerisce il loro utilizzo come fonte di coloranti alimentari dotati di proprietà salutistiche. Obiettivi Caratterizzazione chimica di un estratto concentrato di frutti di P. mahaleb e studio in vitro dei suoi effetti anticancerogeni, antiinfiammatori e antimutagenici. Metodologia I frutti di mahaleb sono stati estratti con solventi consentiti per uso alimentare (etanolo + 1% acido citrico 1M) e l'estratto, concentrato sino a 60 °Brix, analizzato utilizzando metodiche HPLC. Sono stati effettuati saggi in vitro per studiare la capacità dell'estratto di inibire la proliferazione di linee cellulari tumorali umane (MCF-7), le sue proprietà antinfiammatorie su cellule endoteliali vascolari umane e infine i suoi effetti antimutagenici su linee di lievito Saccharomyces cerevisiae D7. Risultati e Discussione L'estratto concentrato ha mostrato un elevato contenuto in antocianine, flavonoli e cumarina ed una elevata attività antiossidante simile o superiore a quella di altri noti estratti concentrati di frutti rossi. Lo studio delle attività biologiche dell'estratto ha mostrato: i) un significativo effetto antiproliferativo e capacità di modulare le GJIC in cellule MCF-7; ii) una inibizione dell'espressione di markers infiammatori tipici dell'attivazione endoteliale (ICAM-1, VCAM-1, E-selectine) in cellule endoteliali ottenute da vena ombelicale umana in coltura; iii) una riduzione di circa tre volte dell'effetto mutageno di H2O2 su cellule di S. cerevisiae. Questi risultati suggeriscono l'utilizzo dei frutti di P. mahaleb processati, come fonte di coloranti alimentari naturali e di composti bioattivi con proprietà salutistiche.

Numerose evidenze scientifiche incoraggiano il consumo di frutta e verdura per prevenire malattie cardiovascolari e tumori. Tuttavia il loro consumo rimane sotto il limite consigliato dall'Organizzazione Mondiale della Sanità. Per questo motivo i consumatori e le aziende alimentari sono alla ricerca di bevande salutistiche "ready-to-drink". I frutti di P. mahaleb, ricchi di composti salutistici (antocianine, flavonoli e cumarine) non sono commestibili a causa del loro sapore amaro, ma l'impiego biotecnologico della fermentazione controllata, utilizzata per migliorare le qualità organolettiche, salutistiche e di serbevolezza di frutta e verdura, potrebbe consentirne l'utilizzo come nuova fonte di probiotici e phytochemicals.

relazione finale

BACKGROUND: The identification of novel plant-based functional foods or nutraceutical ingredients that possess bioactive properties with antioxidant function has recently become important to the food, nutraceutical and cosmetic industries. This study evaluates the polyphenolic composition, identifies bioactive compounds and assays the total antioxidant capacity of Prunus mahaleb L. fruits collected from different populations and sampling years in the countryside around Bari (Apulia Region, Italy). RESULTS: We identified nine polyphenolic compounds including major anthocyanins, coumaric acid derivatives and flavonols from P. mahaleb fruits. The anthocyanin content (in some populations > 500 mg/100 g fresh weight; FW) in the fruit was comparable to that reported for so-called superfruits such as bilberries, chokeberries and blackcurrants. Coumaric acid derivatives comprised a large portion of the total polyphenolic content in the P. mahaleb fruits. Antioxidant activities, assessed using oxygen radical absorbance capacity (ORAC) and Trolox equivalent antioxidant capacity (TEAC) assays, measured up to 15 and 4.5 mmol/100 g FW, respectively. CONCLUSION: Our findings suggest that mahaleb fruit (currently not consumed fresh or used in other ways) could serve as a source of bioactive compounds and therefore find interest from the functional food and nutraceutical industries, as a natural food colorant and antioxidant ingredient in the formulation of functional foods.

L'Artemisia annua L. è una pianta erbacea annuale nota per la produzione del sesquiterpene antimalarico artemisinina. La produzione di artemisinina da parte della pianta è molto bassa e quasi esclusivamente prodotta nelle foglie e nelle infiorescenze, ed accumulata in specifiche strutture (tricomi ghiandolari) (Duke and Paul, 1993; Duke et al., 2009). Pertanto lo sviluppo di biotecnologie per la produzione di artemisinina in laboratorio è di notevole interesse. L'utilizzo di colture in vitro rappresenta una fonte alternativa alla pianta coltivata in campo. Da qui la necessità di sviluppare un valido protocollo per la messa a punto di sistemi in vitro che svincolino la produzione di artemisinina, nella pianta circoscritta ad alcuni organi e limitata a determinati periodi dell'anno, garantendo in questo modo una produzione costante ed efficace nel tempo. Nel nostro laboratorio sono stati allestiti diversi tipi di colture in vitro di A. annua (calli, sospensioni cellulari e piantine micropropagate), ottimizzando le condizioni di crescita mediante l'utilizzo di terreni di coltura con la stessa composizione chimica in macro- e microelementi, ma con ormoni di diverso tipo e concentrazione.

The human health benefits of fruits and vegetables are ascribed to their phytochemical content, such as carotenoids and polyphenolics. In the search for antioxidative chemicals from native fruits of the Puglia region of Italy, Prunus cerasus L., an acidic cherry widely used for culinary purposes, and Prunus mahaleb L., a tree species commonly used as rootstock in cherry crop, were studied. The P. mahaleb fruits have a high content of organic acids, fructose and vitamin C, but are not consumed fresh because of a bitter and sour taste. In this work we obtained the 1H NMR spectra of the two species and from the comparison of these spectra, we found that P. mahaleb fruits have an higher concentration of phenolic compounds, such as flavonoids, and organic acids, in comparison to P. cerasus fruits. The same results were obtained when we focused on anthocyanins. In this study we identified the signals of anthocyanin protons in 1H NMR spectra of a mixture of compounds in aqueous extracts of both P. cerasus and P. mahaleb fruits but the latter species showed a higher concentration and a larger number of these compounds. This metabolomic analysis gave us the data to scientifically revalue traditionally-used plants like P. mahaleb and to identify the potential as source of biofunctional compounds to be used in food and/or pharmaceutical industry. Moreover, in this study, NMR spectroscopy coupled with multivariate data analysis was applied to Prunus metabolomics in order to investigate the botanical origins of Prunus cerasus and to identify the compounds responsible for differentiation of these two species of Prunus (cerasus and mahaleb) and of two cultivars of Prunus cerasus ('Montmorency' and 'Marasca di Zara').

Artemisia annua L., is a herbal plant traditionally used for the treatment of fever. The glandular trichomes of this plant accumulate, although at low levels, artemisinin which is highly effective against malaria. Due to the great importance of this compound many efforts have been made to improve the knowledge on artemisinin production both in plants and in cell cultures. In this study A. annua suspension cultures were established in order to investigate the effects of methyl jasmonate (MeJA) and miconazole treatments on artemisinin biosynthesis. Twenty-two mM MeJA induced a three-fold increase of artemisinin production soon after 30 min. Two hundred mM miconazole induced a 2.5 fold increase of artemisinin production after 24 h treatment but had severe effects on cell viability. The influence of these treatments on the expression of biosynthetic genes was also investigated. MeJA induced an up-regulation of CYP71AV1 while miconazole induced an up-regulation of CPR and DBR2.

Eugenia myrtifolia Sims fruits have been characterized for their anthocyanin content and quality. Following the common procedures and validated methods for separation, identification and quantification of anthocyanins, we found the presence of malvidin 3,5-O-diglucoside as a unique anthocyanin (32,59 mg/100 g fresh weight of the fruit). An in vitro shoot culture of E. myrtifolia was established in order to explore the possibility to produce pigments in vitro. The presence of the only malvidin 3,5-O-diglucoside could be a useful feature in order to manipulate biotechnologically the anthocyanin biosynthetic pathway in the in vitro material (callus and suspension cultures).

Artemisia annua L. è una pianta aromatica erbacea annuale appartenente alla famiglia delle Asteracee, endemica della Cina, ma ormai naturalizzata in Europa e in America. E' oggi coltivata in diversi paesi asiatici per l'estrazione di un composto chiamato artemisisina che risulta molto efficace nel combattere la malaria. L'artemisisina chimicamente è un sesquiterpene lattone caratterizzato da un ponte endoperossidico, al quale viene riconosciuta l'attività antimalarica. Lo scopo delle nostre ricerche è quello di mettere a punto colture in vitro di Artemisia annua L. per la produzione di artemisisina. Sono stati indotti calli da porzioni di foglie incubate al buio in vari mezzi di coltura, tutti con base minerale MS. I risultati preliminari finora ottenuti indicano che il sistema in vitro deve essere migliorato, e per questo pensiamo di utilizzare alcuni precursori della biosintesi del metabolita o di manipolare i parametri fisici e chimici per aumentarne la biosintesi.

In the last years many studies on anthocyanins, the flavonoids responsible for the orange, red and blue colors in different plant organs, have revealed their antioxidant activity and a possible use as chemotherapeutics. The finding that tart cherries (Prunus cerasus L.) contain a great amount of anthocyanins, constituents that possess strong antioxidant and anti-inflammatory activities, have attracted much attention. Here, we report the preliminary results of the induction of anthocyanin biosynthesis in sour cherry callus cell cultures and the characterization of those callus cultures for cell growth and anthocyanin production. Callus cultures have been obtained from the leaf of in vitro grown shoots of two sour cherry cultivars, Amarena Mattarello (A.M.) and Stevnsbaer (S.). Several in vitro culture conditions have been tested to induce the anthocyanin biosynthesis using different media with reduced nitrogen and elevated sucrose contents in order to stimulate anthocyanin accumulation. No stimulatory affect from elevated sucrose concentration was observed, while reduced nitrogen and light seems to be the triggering factors. The pigment production is absent during the first days of growth at the light; afterwards, the pigment accumulation is stimulated by elicitors as light and nutritional factors, and reach a maximum nearly the 20th day of culture, during the exponential phase of growth. The in vitro pigment production system described in cherry callus cultures has given these preliminary but encouraging results, making the plant material very suitable and plastic to the in vitro manipulations.