Innovative treatments for shelf-life extension of whole and fresh-cut fruit

Maintaining the postharvest quality of whole and fresh-cut fruit during storage and distribution is the major challenge facing fruit industry. For this purpose, industry adopt a wide range of technologies to enable extended shelf-life. Many factors can lead to loss of quality in fresh product, hence the common description of these products as ‘perishable’. As a consequence normal factors such as transpiration and respiration lead ultimately to water loss and senescence of the product. Fruits and vegetables are living commodities and their rate of respiration is of key importance to maintenance of quality. It has been commonly observed that the greater the respiration rate of a product, the shorter the shelf-life. The principal problem for fresh-cut fruit industries is the relative shorter shelf-life of minimally processed fruit (MPF) compared to intact product. This fact is strictly connected with the higher ethylene production of fruit tissue stimulated during fresh-cut processing (peeling, cutting, dipping). 1-Methylcyclopropene (1-MCP) is an inhibitor of ethylene action and several researches have shown its effectiveness on the inhibition of ripening and senescence incidence for intact fruit and consequently on their shelf-life extension. More recently 1-MCP treatment has been tested also for shelf-life extension of MPF but discordant results have been obtained. Considering that in some countries 1-MCP is already a commercial product registered for the use on a number of horticultural products, the main aim of this actual study was to enhance our understanding on the effects of 1-MCP treatment on the quality maintenance of whole and fresh-cut climacteric and non-climacteric fruit (apple, kiwifruit and pineapple). Concerning the effects of 1-MCP on whole fruit, was investigated the effects of a semi-commercial postharvest treatment with 1-MCP on the quality of Pink Lady apples as functions of fruit ripening stage, 1-MCP dose, storage time and also in combination with controlled atmospheres storage in order to better understand what is the relationship among these parameters and if is possible to maximize the 1-MCP treatment to meet the market/consumer needs and then in order to put in the market excellent fruit. To achieve this purpose an incomplete three-level three-factor design was adopted. During the storage were monitored several quality parameters: firmness, ripening index, ethylene and carbon dioxide production and were also performed a sensory evaluations after 6 month of storage. In this study the higher retention of firmness (at the end of storage) was achieved by applying the greatest 1-MCP concentration to fruits with the lowest maturity stage. This finding means that in these semi-commercial conditions we may considerate completely blocked the fruit softening. 1-MCP was able to delay also the ethylene and CO2 production and the maturity parameters (soluble solids content and total acidity). Only in some cases 1-MCP generate a synergistic effect with the CA storage. The results of sensory analyses indicated that, the 1-MCP treatment did not affect the sweetness and whole fruit flavour while had a little effect on the decreasing cut fruit flavour. On the contrary the treated apple was more sour, crisp, firm and juicy. The effects of some treatment (dipping and MAP) on the nutrient stability were also investigated showing that in this case study the adopted treatments did not have drastic effects on the antioxidant compounds on the contrary the dipping may enhance the total antioxidant activity by the accumulation of ascorbic acid on the apple cut surface. Results concerning the effects of 1-MCP in combination with MAP on the quality parameters behaviour of the kiwifruit were not always consistent and clear: in terms of colour maintenance, it seemed to have a synergistic effect with N2O MAP; as far as ripening index is concerned, 1-MCP had a preservative effect, but just for sample packed in air.

Il passato nell’epoca della sua (ri)producibilità digitale. Torino 1943 - 45. Metodologia della ricerca con tecnologie informatiche. Sistema storico-territoriale di informazione multimediale

Se il lavoro dello storico è capire il passato come è stato compreso dalla gente che lo ha vissuto, allora forse non è azzardato pensare che sia anche necessario comunicare i risultati delle ricerche con strumenti propri che appartengono a un'epoca e che influenzano la mentalità di chi in quell'epoca vive. Emergenti tecnologie, specialmente nell’area della multimedialità come la realtà virtuale, permettono agli storici di comunicare l’esperienza del passato in più sensi. In che modo la storia collabora con le tecnologie informatiche soffermandosi sulla possibilità di fare ricostruzioni storiche virtuali, con relativi esempi e recensioni? Quello che maggiormente preoccupa gli storici è se una ricostruzione di un fatto passato vissuto attraverso la sua ricreazione in pixels sia un metodo di conoscenza della storia che possa essere considerato valido. Ovvero l'emozione che la navigazione in una realtà 3D può suscitare, è un mezzo in grado di trasmettere conoscenza? O forse l'idea che abbiamo del passato e del suo studio viene sottilmente cambiato nel momento in cui lo si divulga attraverso la grafica 3D? Da tempo però la disciplina ha cominciato a fare i conti con questa situazione, costretta soprattutto dall'invasività di questo tipo di media, dalla spettacolarizzazione del passato e da una divulgazione del passato parziale e antiscientifica. In un mondo post letterario bisogna cominciare a pensare che la cultura visuale nella quale siamo immersi sta cambiando il nostro rapporto con il passato: non per questo le conoscenze maturate fino ad oggi sono false, ma è necessario riconoscere che esiste più di una verità storica, a volte scritta a volte visuale. Il computer è diventato una piattaforma onnipresente per la rappresentazione e diffusione dell’informazione. I metodi di interazione e rappresentazione stanno evolvendo di continuo. Ed è su questi due binari che è si muove l’offerta delle tecnologie informatiche al servizio della storia. Lo scopo di questa tesi è proprio quello di esplorare, attraverso l’utilizzo e la sperimentazione di diversi strumenti e tecnologie informatiche, come si può raccontare efficacemente il passato attraverso oggetti tridimensionali e gli ambienti virtuali, e come, nel loro essere elementi caratterizzanti di comunicazione, in che modo possono collaborare, in questo caso particolare, con la disciplina storica. La presente ricerca ricostruisce alcune linee di storia delle principali fabbriche attive a Torino durante la seconda guerra mondiale, ricordando stretta relazione che esiste tra strutture ed individui e in questa città in particolare tra fabbrica e movimento operaio, è inevitabile addentrarsi nelle vicende del movimento operaio torinese che nel periodo della lotta di Liberazione in città fu un soggetto politico e sociale di primo rilievo. Nella città, intesa come entità biologica coinvolta nella guerra, la fabbrica (o le fabbriche) diventa il nucleo concettuale attraverso il quale leggere la città: sono le fabbriche gli obiettivi principali dei bombardamenti ed è nelle fabbriche che si combatte una guerra di liberazione tra classe operaia e autorità, di fabbrica e cittadine. La fabbrica diventa il luogo di "usurpazione del potere" di cui parla Weber, il palcoscenico in cui si tengono i diversi episodi della guerra: scioperi, deportazioni, occupazioni .... Il modello della città qui rappresentata non è una semplice visualizzazione ma un sistema informativo dove la realtà modellata è rappresentata da oggetti, che fanno da teatro allo svolgimento di avvenimenti con una precisa collocazione cronologica, al cui interno è possibile effettuare operazioni di selezione di render statici (immagini), di filmati precalcolati (animazioni) e di scenari navigabili interattivamente oltre ad attività di ricerca di fonti bibliografiche e commenti di studiosi segnatamente legati all'evento in oggetto. Obiettivo di questo lavoro è far interagire, attraverso diversi progetti, le discipline storiche e l’informatica, nelle diverse opportunità tecnologiche che questa presenta. Le possibilità di ricostruzione offerte dal 3D vengono così messe a servizio della ricerca, offrendo una visione integrale in grado di avvicinarci alla realtà dell’epoca presa in considerazione e convogliando in un’unica piattaforma espositiva tutti i risultati. Divulgazione Progetto Mappa Informativa Multimediale Torino 1945 Sul piano pratico il progetto prevede una interfaccia navigabile (tecnologia Flash) che rappresenti la pianta della città dell’epoca, attraverso la quale sia possibile avere una visione dei luoghi e dei tempi in cui la Liberazione prese forma, sia a livello concettuale, sia a livello pratico. Questo intreccio di coordinate nello spazio e nel tempo non solo migliora la comprensione dei fenomeni, ma crea un maggiore interesse sull’argomento attraverso l’utilizzo di strumenti divulgativi di grande efficacia (e appeal) senza perdere di vista la necessità di valicare le tesi storiche proponendosi come piattaforma didattica. Un tale contesto richiede uno studio approfondito degli eventi storici al fine di ricostruire con chiarezza una mappa della città che sia precisa sia topograficamente sia a livello di navigazione multimediale. La preparazione della cartina deve seguire gli standard del momento, perciò le soluzioni informatiche utilizzate sono quelle fornite da Adobe Illustrator per la realizzazione della topografia, e da Macromedia Flash per la creazione di un’interfaccia di navigazione. La base dei dati descrittivi è ovviamente consultabile essendo contenuta nel supporto media e totalmente annotata nella bibliografia. È il continuo evolvere delle tecnologie d'informazione e la massiccia diffusione dell’uso dei computer che ci porta a un cambiamento sostanziale nello studio e nell’apprendimento storico; le strutture accademiche e gli operatori economici hanno fatto propria la richiesta che giunge dall'utenza (insegnanti, studenti, operatori dei Beni Culturali) di una maggiore diffusione della conoscenza storica attraverso la sua rappresentazione informatizzata. Sul fronte didattico la ricostruzione di una realtà storica attraverso strumenti informatici consente anche ai non-storici di toccare con mano quelle che sono le problematiche della ricerca quali fonti mancanti, buchi della cronologia e valutazione della veridicità dei fatti attraverso prove. Le tecnologie informatiche permettono una visione completa, unitaria ed esauriente del passato, convogliando tutte le informazioni su un'unica piattaforma, permettendo anche a chi non è specializzato di comprendere immediatamente di cosa si parla. Il miglior libro di storia, per sua natura, non può farlo in quanto divide e organizza le notizie in modo diverso. In questo modo agli studenti viene data l'opportunità di apprendere tramite una rappresentazione diversa rispetto a quelle a cui sono abituati. La premessa centrale del progetto è che i risultati nell'apprendimento degli studenti possono essere migliorati se un concetto o un contenuto viene comunicato attraverso più canali di espressione, nel nostro caso attraverso un testo, immagini e un oggetto multimediale. Didattica La Conceria Fiorio è uno dei luoghi-simbolo della Resistenza torinese. Il progetto è una ricostruzione in realtà virtuale della Conceria Fiorio di Torino. La ricostruzione serve a arricchire la cultura storica sia a chi la produce, attraverso una ricerca accurata delle fonti, sia a chi può poi usufruirne, soprattutto i giovani, che, attratti dall’aspetto ludico della ricostruzione, apprendono con più facilità. La costruzione di un manufatto in 3D fornisce agli studenti le basi per riconoscere ed esprimere la giusta relazione fra il modello e l’oggetto storico. Le fasi di lavoro attraverso cui si è giunti alla ricostruzione in 3D della Conceria: . una ricerca storica approfondita, basata sulle fonti, che possono essere documenti degli archivi o scavi archeologici, fonti iconografiche, cartografiche, ecc.; . La modellazione degli edifici sulla base delle ricerche storiche, per fornire la struttura geometrica poligonale che permetta la navigazione tridimensionale; . La realizzazione, attraverso gli strumenti della computer graphic della navigazione in 3D. Unreal Technology è il nome dato al motore grafico utilizzato in numerosi videogiochi commerciali. Una delle caratteristiche fondamentali di tale prodotto è quella di avere uno strumento chiamato Unreal editor con cui è possibile costruire mondi virtuali, e che è quello utilizzato per questo progetto. UnrealEd (Ued) è il software per creare livelli per Unreal e i giochi basati sul motore di Unreal. E’ stata utilizzata la versione gratuita dell’editor. Il risultato finale del progetto è un ambiente virtuale navigabile raffigurante una ricostruzione accurata della Conceria Fiorio ai tempi della Resistenza. L’utente può visitare l’edificio e visualizzare informazioni specifiche su alcuni punti di interesse. La navigazione viene effettuata in prima persona, un processo di “spettacolarizzazione” degli ambienti visitati attraverso un arredamento consono permette all'utente una maggiore immersività rendendo l’ambiente più credibile e immediatamente codificabile. L’architettura Unreal Technology ha permesso di ottenere un buon risultato in un tempo brevissimo, senza che fossero necessari interventi di programmazione. Questo motore è, quindi, particolarmente adatto alla realizzazione rapida di prototipi di una discreta qualità, La presenza di un certo numero di bug lo rende, però, in parte inaffidabile. Utilizzare un editor da videogame per questa ricostruzione auspica la possibilità di un suo impiego nella didattica, quello che le simulazioni in 3D permettono nel caso specifico è di permettere agli studenti di sperimentare il lavoro della ricostruzione storica, con tutti i problemi che lo storico deve affrontare nel ricreare il passato. Questo lavoro vuole essere per gli storici una esperienza nella direzione della creazione di un repertorio espressivo più ampio, che includa gli ambienti tridimensionali. Il rischio di impiegare del tempo per imparare come funziona questa tecnologia per generare spazi virtuali rende scettici quanti si impegnano nell'insegnamento, ma le esperienze di progetti sviluppati, soprattutto all’estero, servono a capire che sono un buon investimento. Il fatto che una software house, che crea un videogame di grande successo di pubblico, includa nel suo prodotto, una serie di strumenti che consentano all'utente la creazione di mondi propri in cui giocare, è sintomatico che l'alfabetizzazione informatica degli utenti medi sta crescendo sempre più rapidamente e che l'utilizzo di un editor come Unreal Engine sarà in futuro una attività alla portata di un pubblico sempre più vasto. Questo ci mette nelle condizioni di progettare moduli di insegnamento più immersivi, in cui l'esperienza della ricerca e della ricostruzione del passato si intreccino con lo studio più tradizionale degli avvenimenti di una certa epoca. I mondi virtuali interattivi vengono spesso definiti come la forma culturale chiave del XXI secolo, come il cinema lo è stato per il XX. Lo scopo di questo lavoro è stato quello di suggerire che vi sono grosse opportunità per gli storici impiegando gli oggetti e le ambientazioni in 3D, e che essi devono coglierle. Si consideri il fatto che l’estetica abbia un effetto sull’epistemologia. O almeno sulla forma che i risultati delle ricerche storiche assumono nel momento in cui devono essere diffuse. Un’analisi storica fatta in maniera superficiale o con presupposti errati può comunque essere diffusa e avere credito in numerosi ambienti se diffusa con mezzi accattivanti e moderni. Ecco perchè non conviene seppellire un buon lavoro in qualche biblioteca, in attesa che qualcuno lo scopra. Ecco perchè gli storici non devono ignorare il 3D. La nostra capacità, come studiosi e studenti, di percepire idee ed orientamenti importanti dipende spesso dai metodi che impieghiamo per rappresentare i dati e l’evidenza. Perché gli storici possano ottenere il beneficio che il 3D porta con sè, tuttavia, devono sviluppare un’agenda di ricerca volta ad accertarsi che il 3D sostenga i loro obiettivi di ricercatori e insegnanti. Una ricostruzione storica può essere molto utile dal punto di vista educativo non sono da chi la visita ma, anche da chi la realizza. La fase di ricerca necessaria per la ricostruzione non può fare altro che aumentare il background culturale dello sviluppatore. Conclusioni La cosa più importante è stata la possibilità di fare esperienze nell’uso di mezzi di comunicazione di questo genere per raccontare e far conoscere il passato. Rovesciando il paradigma conoscitivo che avevo appreso negli studi umanistici, ho cercato di desumere quelle che potremo chiamare “leggi universali” dai dati oggettivi emersi da questi esperimenti. Da punto di vista epistemologico l’informatica, con la sua capacità di gestire masse impressionanti di dati, dà agli studiosi la possibilità di formulare delle ipotesi e poi accertarle o smentirle tramite ricostruzioni e simulazioni. Il mio lavoro è andato in questa direzione, cercando conoscere e usare strumenti attuali che nel futuro avranno sempre maggiore presenza nella comunicazione (anche scientifica) e che sono i mezzi di comunicazione d’eccellenza per determinate fasce d’età (adolescenti). Volendo spingere all’estremo i termini possiamo dire che la sfida che oggi la cultura visuale pone ai metodi tradizionali del fare storia è la stessa che Erodoto e Tucidide contrapposero ai narratori di miti e leggende. Prima di Erodoto esisteva il mito, che era un mezzo perfettamente adeguato per raccontare e dare significato al passato di una tribù o di una città. In un mondo post letterario la nostra conoscenza del passato sta sottilmente mutando nel momento in cui lo vediamo rappresentato da pixel o quando le informazioni scaturiscono non da sole, ma grazie all’interattività con il mezzo. La nostra capacità come studiosi e studenti di percepire idee ed orientamenti importanti dipende spesso dai metodi che impieghiamo per rappresentare i dati e l’evidenza. Perché gli storici possano ottenere il beneficio sottinteso al 3D, tuttavia, devono sviluppare un’agenda di ricerca volta ad accertarsi che il 3D sostenga i loro obiettivi di ricercatori e insegnanti. Le esperienze raccolte nelle pagine precedenti ci portano a pensare che in un futuro non troppo lontano uno strumento come il computer sarà l’unico mezzo attraverso cui trasmettere conoscenze, e dal punto di vista didattico la sua interattività consente coinvolgimento negli studenti come nessun altro mezzo di comunicazione moderno.

Studio molecolare dei meccanismi dell'autoincompatibilità gametofitica in pero europeo (Pyrus communis)

Self-incompatibility (SI) systems have evolved in many flowering plants to prevent self-fertilization and thus promote outbreeding. Pear and apple, as many of the species belonging to the Rosaceae, exhibit RNase-mediated gametophytic self-incompatibility, a widespread system carried also by the Solanaceae and Plantaginaceae. Pear orchards must for this reason contain at least two different cultivars that pollenize each other; to guarantee an efficient cross-pollination, they should have overlapping flowering periods and must be genetically compatible. This compatibility is determined by the S-locus, containing at least two genes encoding for a female (pistil) and a male (pollen) determinant. The female determinant in the Rosaceae, Solanaceae and Plantaginaceae system is a stylar glycoprotein with ribonuclease activity (S-RNase), that acts as a specific cytotoxin in incompatible pollen tubes degrading cellular RNAs. Since its identification, the S-RNase gene has been intensively studied and the sequences of a large number of alleles are available in online databases. On the contrary, the male determinant has been only recently identified as a pollen-expressed protein containing a F-box motif, called S-Locus F-box (abbreviated SLF or SFB). Since F-box proteins are best known for their participation to the SCF (Skp1 - Cullin - F-box) E3 ubiquitine ligase enzymatic complex, that is involved in protein degradation through the 26S proteasome pathway, the male determinant is supposed to act mediating the ubiquitination of the S-RNases, targeting them for the degradation in compatible pollen tubes. Attempts to clone SLF/SFB genes in the Pyrinae produced no results until very recently; in apple, the use of genomic libraries allowed the detection of two F-box genes linked to each S haplotype, called SFBB (S-locus F-Box Brothers). In Japanese pear, three SFBB genes linked to each haplotype were cloned from pollen cDNA. The SFBB genes exhibit S haplotype-specific sequence divergence and pollen-specific expression; their multiplicity is a feature whose interpretation is unclear: it has been hypothesized that all of them participate in the S-specific interaction with the RNase, but it is also possible that only one of them is involved in this function. Moreover, even if the S locus male and female determinants are the only responsible for the specificity of the pollen-pistil recognition, many other factors are supposed to play a role in GSI; these are not linked to the S locus and act in a S-haplotype independent manner. They can have a function in regulating the expression of S determinants (group 1 factors), modulating their activity (group 2) or acting downstream, in the accomplishment of the reaction of acceptance or rejection of the pollen tube (group 3). This study was aimed to the elucidation of the molecular mechanism of GSI in European pear (Pyrus communis) as well as in the other Pyrinae; it was divided in two parts, the first focusing on the characterization of male determinants, and the second on factors external to the S locus. The research of S locus F-box genes was primarily aimed to the identification of such genes in European pear, for which sequence data are still not available; moreover, it allowed also to investigate about the S locus structure in the Pyrinae. The analysis was carried out on a pool of varieties of the three species Pyrus communis (European pear), Pyrus pyrifolia (Japanese pear), and Malus × domestica (apple); varieties carrying S haplotypes whose RNases are highly similar were chosen, in order to check whether or not the same level of similarity is maintained also between the male determinants. A total of 82 sequences was obtained, 47 of which represent the first S-locus F-box genes sequenced from European pear. The sequence data strongly support the hypothesis that the S locus structure is conserved among the three species, and presumably among all the Pyrinae; at least five genes have homologs in the analysed S haplotypes, but the number of F-box genes surrounding the S-RNase could be even greater. The high level of sequence divergence and the similarity between alleles linked to highly conserved RNases, suggest a shared ancestral polymorphism also for the F-box genes. The F-box genes identified in European pear were mapped on a segregating population of 91 individuals from the cross 'Abbé Fétel' × 'Max Red Bartlett'. All the genes were placed on the linkage group 17, where the S locus has been placed both in pear and apple maps, and resulted strongly associated to the S-RNase gene. The linkage with the RNase was perfect for some of the F-box genes, while for others very rare single recombination events were identified. The second part of this study was focused on the research of other genes involved in the SI response in pear; it was aimed on one side to the identification of genes differentially expressed in compatible and incompatible crosses, and on the other to the cloning and characterization of the transglutaminase (TGase) gene, whose role may be crucial in pollen rejection. For the identification of differentially expressed genes, controlled pollinations were carried out in four combinations (self pollination, incompatible, half-compatible and fully compatible cross-pollination); expression profiles were compared through cDNA-AFLP. 28 fragments displaying an expression pattern related to compatibility or incompatibility were identified, cloned and sequenced; the sequence analysis allowed to assign a putative annotation to a part of them. The identified genes are involved in very different cellular processes or in defense mechanisms, suggesting a very complex change in gene expression following the pollen/pistil recognition. The pool of genes identified with this technique offers a good basis for further study toward a better understanding of how the SI response is carried out. Among the factors involved in SI response, moreover, an important role may be played by transglutaminase (TGase), an enzyme involved both in post-translational protein modification and in protein cross-linking. The TGase activity detected in pear styles was significantly higher when pollinated in incompatible combinations than in compatible ones, suggesting a role of this enzyme in the abnormal cytoskeletal reorganization observed during pollen rejection reaction. The aim of this part of the work was thus to identify and clone the pear TGase gene; the PCR amplification of fragments of this gene was achieved using primers realized on the alignment between the Arabidopsis TGase gene sequence and several apple EST fragments; the full-length coding sequence of the pear TGase gene was then cloned from cDNA, and provided a precious tool for further study of the in vitro and in vivo action of this enzyme.

Influence of climate changes on cross allergies: possible involvement of pollen transglutaminase

Allergies are a complex of symptoms derived from altered IgE-mediated reactions of the immune system towards substances known as allergens. Allergic sensibilization can be of food or respiratory origin and, in particular, apple and hazelnut allergens have been identified in pollens or fruits. Allergic cross-reactivity can occur in a patient reacting to similar allergens from different origins, justifying the research in both systems as in Europe a greater number of people suffers from apple fruit allergy, but little evidence exists about pollen. Apple fruit allergies are due to four different classes of allergens (Mal d 1, 2, 3, 4), whose allergenicity is related both to genotype and tissue specificity; therefore I have investigated their presence also in pollen at different time of germination to clarify the apple pollen allergenic potential. I have observed that the same four classes of allergens found in fruit are expressed at different levels also in pollen, and their presence might support that the apple pollen can be considered allergenic as the fruit, deducing that apple allergy could also be indirectly caused by sensitization to pollen. Climate changes resulting from increases in temperature and air pollution influence pollen allergenicity, responsible for the dramatic raise in respiratory allergies (hay fever, bronchial asthma, conjunctivitis). Although the link between climate change and pollen allergenicity is proven, the underlying mechanism is little understood. Transglutaminases (TGases), a class of enzymes able to post-translationally modify proteins, are activated under stress and involved in some inflammatory responses, enhancing the activity of pro-inflammatory phospholipase A2, suggesting a role in allergies. Recently, a calcium-dependent TGase activity has been identified in the pollen cell wall, raising the possibility that pollen TGase may have a role in the modification of pollen allergens reported above, thus stabilizing them against proteases. This enzyme can be involved also in the transamidation of proteins present in the human mucosa interacting with surface pollen or, finally, the enzyme itself can represent an allergen, as suggested by studies on celiac desease. I have hypothesized that this pollen enzyme can be affected by climate changes and be involved in exhacerbating allergy response. The data presented in this thesis represent a scientific basis for future development of studies devoted to verify the hypothesis set out here. First, I have demonstrated the presence of an extracellular TGase on the surface of the grain observed either at the apical or the proximal parts of the pollen-tube by laser confocal microscopy (Iorio et al., 2008), that plays an essential role in apple pollen-tube growth, as suggested by the arrest of tube elongation by TGase inhibitors, such as EGTA or R281. Its involvement in pollen tube growth is mainly confirmed by the data of activity and gene expression, because TGase showed a peak between 15 min and 30 min of germination, when this process is well established, and an optimal pH around 6.5, which is close to that recorded for the germination medium. Moreover, data show that pollen TGase can be a glycoprotein as the glycosylation profile is linked both with the activation of the enzyme and with its localization at the pollen cell wall during germination, because from the data presented seems that the active form of TGase involved in pollen tube growth and pollen-stylar interaction is more exposed and more weakly bound to the cell wall. Interestingly, TGase interacts with fibronectin (FN), a putative SAMs or psECM component, inducing possibly intracellular signal transduction during the interaction between pollen-stylar occuring in the germination process, since a protein immunorecognised by anti-FN antibody is also present in pollen, in particular at the level of pollen grain cell wall in a punctuate pattern, but also along the shank of the pollen tube wall, in a similar pattern that recalls the signal obtained with the antibody anti TGase. FN represents a good substrate for the enzyme activity, better than DMC usually used as standard substrate for animal TGase. Thus, this pollen enzyme, necessary for its germination, is exposed on the pollen surface and consequently can easily interact with mucosal proteins, as it has been found germinated pollen in studies conducted on human mucus (Forlani, personal communication). I have obtained data that TGase activity increases in a very remarkable way when pollen is exposed to stressful conditions, such as climate changes and environmental pollution. I have used two different species of pollen, an aero allergenic (hazelnut, Corylus avellana) pollen, whose allergenicity is well documented, and an enthomophylus (apple, Malus domestica) pollen, which is not yet well characterized, to compare data on their mechanism of action in response to stressors. The two pollens have been exposed to climate changes (different temperatures, relative humidity (rH), acid rain at pH 5.6 and copper pollution (3.10 µg/l)) and showed an increase in pollen surface TGase activity that is not accompanied to an induced expression of TGase immunoreactive protein with AtPNG1p. Probably, climate change induce an alteration or damage to pollen cell wall that carries the pollen grains to release their content in the medium including TGase enzyme, that can be free to carry out its function as confirmed by the immunolocalisation and by the in situ TGase activity assay data; morphological examination indicated pollen damage, viability significantly reduced and in acid rain conditions an early germination of apple pollen, thus possibly enhancing the TGase exposure on pollen surface. Several pollen proteins were post-translationally modified, as well as mammalian sPLA2 especially with Corylus pollen, which results in its activation, potentially altering pollen allergenicity and inflammation. Pollen TGase activity mimicked the behaviour of gpl TGase and AtPNG1p in the stimulation of sPLA2, even if the regulatory mechanism seems different to gpl TGase, because pollen TGase favours an intermolecular cross-linking between various molecules of sPLA2, giving rise to high-molecular protein networks normally more stable. In general, pollens exhibited a significant endogenous phospholipase activity and it has been observed differences according to the allergenic (Corylus) or not-well characterized allergenic (Malus) attitude of the pollen. However, even if with a different intensity level in activation, pollen enzyme share the ability to activate the sPLA2, thus suggesting an important regulatory role for the activation of a key enzyme of the inflammatory response, among which my interest was addressed to pollen allergy. In conclusion, from all the data presented, mainly presence of allergens, presence of an extracellular TGase, increasing in its activity following exposure to environmental pollution and PLA2 activation, I can conclude that also Malus pollen can behave as potentially allergenic. The mechanisms described here that could affect the allergenicity of pollen, maybe could be the same occurring in fruit, paving the way for future studies in the identification of hyper- and hypo- allergenic cultivars, in preventing environmental stressor effects and, possibly, in the production of transgenic plants.

Advances in understanding of osmotic dehydration and vacuum impregnation of fruits

Osmotic Dehydration and Vacuum Impregnation are interesting operations in the food industry with applications in minimal fruit processing and/or freezing, allowing to develop new products with specific innovative characteristics. Osmotic dehydration is widely used for the partial removal of water from cellular tissue by immersion in hypertonic (osmotic) solution. The driving force for the diffusion of water from the tissue is provided by the differences in water chemical potential between the external solution and the internal liquid phase of the cells. Vacuum Impregnation of porous products immersed in a liquid phase consist of reduction of pressure in a solid-liquid system (vacuum step) followed by the restoration of atmospheric pressure (atmospheric step). During the vacuum step the internal gas in the product pores is expanded and partially flows out while during the atmospheric step, there is a compression of residual gas and the external liquid flows into the pores (Fito, 1994). This process is also a very useful unit operation in food engineering as it allows to introduce specific solutes in the tissue which can play different functions (antioxidants, pH regulators, preservatives, cryoprotectants etc.). The present study attempts to enhance our understanding and knowledge of fruit as living organism, interacting dynamically with the environment, and to explore metabolic, structural, physico-chemical changes during fruit processing. The use of innovative approaches and/or technologies such as SAFES (Systematic Approach to Food Engineering System), LF-NMR (Low Frequency Nuclear Magnetic Resonance), GASMAS (Gas in Scattering Media Absorption Spectroscopy) are very promising to deeply study these phenomena. SAFES methodology was applied in order to study irreversibility of the structural changes of kiwifruit during short time of osmotic treatment. The results showed that the deformed tissue can recover its initial state 300 min after osmotic dehydration at 25 °C. The LF-NMR resulted very useful in water status and compartmentalization study, permitting to separate observation of three different water population presented in vacuole, cytoplasm plus extracellular space and cell wall. GASMAS techniques was able to study the pressure equilibration after Vacuum Impregnation showing that after restoration of atmospheric pressure in the solid-liquid system, there was a reminding internal low pressure in the apple tissue that slowly increases until reaching the atmospheric pressure, in a time scale that depends on the vacuum applied during the vacuum step. The physiological response of apple tissue on Vacuum Impregnation process was studied indicating the possibility of vesicular transport within the cells. Finally, the possibility to extend the freezing tolerance of strawberry fruits impregnated with cryoprotectants was proven.

Influence of nitrogen and soil physical characteristics on belowground carbon flux dynamics of woody plants

At ecosystem level soil respiration (Rs) represents the largest carbon (C) flux after gross primary productivity, being mainly generated by root respiration (autotrophic respiration, Ra) and soil microbial respiration (heterotrophic respiration, Rh). In the case of terrestrial ecosystems, soils contain the largest C-pool, storing twice the amount of C contained in plant biomass. Soil organic matter (SOM), representing the main C storage in soil, is decomposed by soil microbial community. This process produces CO2 which is mainly released as Rh. It is thus relevant to understand how microbial activity is influenced by environmental factors like soil temperature, soil moisture and nutrient availability, since part of the CO2 produced by Rh, directly increases atmospheric CO2 concentration and therefore affects the phenomenon of climate change. Among terrestrial ecosystems, agricultural fields have traditionally been considered as sources of atmospheric CO2. In agricultural ecosystems, in particular apple orchards, I identified the role of root density, soil temperature, soil moisture and nitrogen (N) availability on Rs and on its two components, Ra and Rh. To do so I applied different techniques to separate Rs in its two components, the ”regression technique” and the “trenching technique”. I also studied the response of Ra to different levels of N availability, distributed either in a uniform or localized way, in the case of Populus tremuloides trees. The results showed that Rs is mainly driven by soil temperature, to which it is positively correlated, that high levels of soil moisture have inhibiting effects, and that N has a negligible influence on total Rs, as well as on Ra. Further I found a negative response of Rh to high N availability, suggesting that microbial decomposition processes in the soil are inhibited by the presence of N. The contribution of Ra to Rs was of 37% on average.

Development of instrumental and sensory analytical methods of food obtained by traditional and emerging technologies

The consumer demand for natural, minimally processed, fresh like and functional food has lead to an increasing interest in emerging technologies. The aim of this PhD project was to study three innovative food processing technologies currently used in the food sector. Ultrasound-assisted freezing, vacuum impregnation and pulsed electric field have been investigated through laboratory scale systems and semi-industrial pilot plants. Furthermore, analytical and sensory techniques have been developed to evaluate the quality of food and vegetable matrix obtained by traditional and emerging processes. Ultrasound was found to be a valuable technique to improve the freezing process of potatoes, anticipating the beginning of the nucleation process, mainly when applied during the supercooling phase. A study of the effects of pulsed electric fields on phenol and enzymatic profile of melon juice has been realized and the statistical treatment of data was carried out through a response surface method. Next, flavour enrichment of apple sticks has been realized applying different techniques, as atmospheric, vacuum, ultrasound technologies and their combinations. The second section of the thesis deals with the development of analytical methods for the discrimination and quantification of phenol compounds in vegetable matrix, as chestnut bark extracts and olive mill waste water. The management of waste disposal in mill sector has been approached with the aim of reducing the amount of waste, and at the same time recovering valuable by-products, to be used in different industrial sectors. Finally, the sensory analysis of boiled potatoes has been carried out through the development of a quantitative descriptive procedure for the study of Italian and Mexican potato varieties. An update on flavour development in fresh and cooked potatoes has been realized and a sensory glossary, including general and specific definitions related to organic products, used in the European project Ecropolis, has been drafted.

Phenotypic and molecular characterization of ethylene biosynthesis in apples

Ethylene plays an important role in apple fruit development. Its biosynthesis is catalyzed by two enzymes ACS and ACO. The first is considered to catalyzes the rate-limiting step of ethylene production and in apple two different alleles (MdACS1-1 and MdACS1-2) of this gene have been identified. The presence in the promoter region of MdACS1-2 allele of a SINE insertion is considered to be responsible for a low transcription level and a pronounced reduction in ethylene production in apple cultivar homozygous for this allele. However, the specific expression of each MdACS1 allele has never been reported as well as any in vivo analysis of its 5’-flanking region. With the present study we addressed these issues by developing a set of qPCR allele specific primers for MdACS1 and by a functional characterization of the MdACS1 promoters by transient expression analysis. qPCR analysis on different apple tissues and stages of development demonstrated that MdACS1-2 allele is never express and that MdACS1-1 allele is ripening-related and expresses predominantly but not exclusively in apple fruit. To test MdACS1 promoter in fruit the only protocol available in literature for transient transformation of apple fruit was evaluated and optimized. Twenty chimeric promoter::reporter constructs were generated and analyzed by Agrobacterium-transient transformation. The in vivo analysis allowed to identify an enhancer-like region of 261 bp in MdACS1 promoter and a region of 57 bp in MdACS1-2 responsible, also if not alone, in the inactivation of the MdACS1-2 allele. Through the assessment of ethylene production in a segregating progeny derived from the cross between Fuji and Mondial Gala (homozygous for MdACS1-2 allele) we demonstrated that at least two other genes may be involved in apple ethylene production. An hypothesis that could explain the difference between Fuji and Mondial Gala have been proposed.

Phenotype and genotype characterization of Monilinia spp. isolates and preformed antifungal compounds in peach peel fruit at different developmental stages

The brown rot fungi belong to a group of fungal pathogens that causes considerable damage to cultivated fruits trees, particularly stone fruits and apples in the temperate regions of the World and during the postharvest with an important economic impact. In particular in Italy, it is important to monitor the Monilinia population to control economic losses associated to the peach and nectarine market. This motivates the research steps presented in this dissertation on Monilinia Italian isolates. The Monilinia species collected from stone fruits have been identified using molecular analysis based on specific primers. The relevant role of M. fructicola was confirmed and, for the first time, it was found also on apple fruits. To avoid the development of resistant strains and implement valid treatment strategies, the understanding of the fruit natural resistance during different developmental stages and the assessment of the Monilinia sensitivity/resistance to fungicides are required. The relationship between the inhibition spots and the phenolic compounds in peach fruit peel was highlighted in this research. Three methods were used to assess isolate resistance/sensitivity, the amended medium, the Spiral Gradient Endpoint Method (SGD) and the Alamar Blue method. The PCR was used to find possible mutation points in the b-tubulin gene that is responsible for fungicide resistance. Interestingly, no mutation points were observed in resistant M. laxa isolates, suggesting that the resistance could be stimulated by environmental factors. This lead to the study of the effect of the temperature on the resistance and the preliminary results of in vitro tests showed that maximum inhibition was observed at 30°C.

Le opere di urbanizzazione a scomputo degli oneri

Le considerazioni sviluppate in questo scritto si pongono come obiettivo quello di fare chiarezza sul delicato tema delle opere di urbanizzazione a scomputo. La normativa concernente la realizzazione delle opere pubbliche a scomputo totale o parziale degli oneri di urbanizzazione è stata oggetto di svariate modifiche e interpretazioni giurisprudenziali, che si sono susseguite dopo l'importante pronuncia della Corte di Giustizia Europea. E' con questa sentenza che i Giudici del Kirchberg introducono un particolare obbligo procedurale a carico dei privati: nel caso in cui singole opere superino i valori di rilevanza europea, esse devono essere affidate, applicando le procedure di gara previste dalla direttiva 37/93/CEE. Va precisato che sino a quel momento l'affidamento diretto delle opere al privato costituiva nell'ottica del Legislatore lo strumento per realizzare le infrastrutture necessarie per consentire gli insediamenti edilizi che la pubblica amministrazione spesso non era in grado di effettuare. In questo panorama legislativo la sentenza della Corte di Giustizia, appare del tutto dirompente. Infatti, introducendo il principio secondo cui anche la realizzazione diretta delle opere di urbanizzazione da parte del privato deve sottostare alle regole delle procedure europee in materia di appalti, mette inevitabilmente a confronto due normative, quella degli appalti pubblici e quella dell'urbanistica, che sino a quel momento erano riuscite a viaggiare in modo parallelo, senza dar luogo a reciproche sovrapposizioni. Il Legislatore nazionale ha, con molta fatica, recepito il principio comunitario ed è stato negli anni quasi costretto, attraverso una serie di modifiche legislative, ad ampliarne la portata. La presente ricerca, dopo aver analizzato i vari correttivi apportati al Codice degli appalti pubblici vuole, quindi, verificare se l'attuale quadro normativo rappresenti un vero punto di equilibrio tra le contrapposte esigenze di pianificazione del territorio e di rispetto dei principi comunitari di concorrenza nella scelta del contraente.

Control of fruit postharvest diseases by thermotherapy: understanding the mechanisms of action by –omics approaches

In recent years the hot water treatment (HW) represents an effective and safe approach for managing postharvest decay. This study reported the effect of an HW (60°C for 60 s and 45°C for 10 min) on brown rot and blue mould respectively. Peaches was found more thermotolerant compared to apple fruit, otherwise Penicillium expansum was more resistant to heat with respect to Monilinia spp. In semi-commercial and commercial trials, the inhibition of brown rot in naturally infected peaches was higher than 78% after 6 days at 0°C and 3 days at 20°C. Moreover, in laboratory trials a 100% disease incidence reduction was obtained by treating artificially infected peaches at 6-12 h after inoculation revealing a curative effect of HW. The expression levels of some genes were evaluated by qRT-PCR. Specifically, the cell wall genes (β-GAL, PL, PG, PME) showed a general decrease of expression level whereas PAL, CHI, HSP70 and ROS-scavenging genes were induced in treated peaches compared to the control ones. Contrarily, HW applied on artificially infected fruit before the inoculum was found to increase brown rot susceptibility. This aspect might be due to an increase of fruit VOCs emission as revealed by PTR-ToF-MS analysis. In addition a microarray experiment was conducted to analyze molecular mechanisms underneath the apple response to heat. Our results showed a largest amount of induced Heat shock proteins (HSPs), Heat shock cognate proteins (HSCs), Heat shock transcription factors (HSTFs) genes found at 1 and 4 hours from the treatment. Those genes required for the thermotolerance process could be involved in induced resistance response. The hypothesis was confirmed by 30% of blue mold disease reduction in artificially inoculated apple after 1 and 4 hours from the treatment. In order to improve peaches quality and disease management during storage, an innovative tool was also used: Da-meter.

Identification and genetic diversity in phytoplasmas associated with diseases of cassava and other agronomic relevant crops in south-east Asia and Latin America

Identification and genetic diversity of phytoplasmas infecting tropical plant species, selected among those most agronomically relevant in South-east Asia and Latin America were studied. Correlation between evolutionary divergence of relevant phytoplasma strains and their geographic distribution by comparison on homologous genes of phytoplasma strains detected in the same or related plant species in other geographical areas worldwide was achieved. Molecular diversity was studied on genes coding ribosomal proteins, groEL, tuf and amp besides phytoplasma 16S rRNA. Selected samples infected by phytoplasmas belonging to diverse ribosomal groups were also studied by in silico RFLP followed by phylogenetic analyses. Moreover a partial genome annotation of a ‘Ca. P. brasiliense’ strain was done towards future application for epidemiological studies. Phytoplasma presence in cassava showing frog skin (CFSD) and witches’ broom (CWB) diseases in Costa Rica - Paraguay and in Vietnam – Thailand, respectively, was evaluated. In both cases, the diseases were associated with phytoplasmas related to aster yellows, apple proliferation and “stolbur” groups, while only phytoplasma related to X-disease group in CFSD, and to hibiscus witches’ broom, elm yellows and clover proliferation groups in CWB. Variability was found among strains belonging to the same ribosomal group but having different geographic origin and associated with different disease. Additionally, a dodder transmission assay to elucidate the role of phytoplasmas in CWB disease was carried out, and resulted in typical phytoplasma symptoms in periwinkle plants associated with the presence of aster yellows-related strains. Lethal wilt disease, a severe disease of oil palm in Colombia that is spreading throughout South America was also studied. Phytoplasmas were detected in symptomatic oil palm and identified as ‘Ca. P. asteris’, ribosomal subgroup 16SrI-B, and were distinguished from other aster yellows phytoplasmas used as reference strains; in particular, from an aster yellows strain infecting corn in the same country.

Sensory and instrumental profiling of apples: a new tool for quality assessment

Food suppliers currently measure apple quality considering basic pomological descriptors. Sensory analysis is expensive, does not permit to analyse many samples, and cannot be implemented for measuring quality properties in real time. However, sensory analysis is the best way to precisely describe food eating quality, since it is able to define, measure, and explain what is really perceivable by human senses and using a language that closely reflects the consumers’ perception. On the basis of such observations, we developed a detailed protocol for apple sensory profiling by descriptive sensory analysis and instrumental measurements. The collected sensory data were validated by applying rigorous scientific criteria for sensory analysis. The method was then applied for studying sensory properties of apples and their changes in relation to different pre- and post-harvest factors affecting fruit quality, and demonstrated to be able to discriminate fruit varieties and to highlight differences in terms of sensory properties. The instrumental measurements confirmed such results. Moreover, the correlation between sensory and instrumental data was studied, and a new effective approach was defined for the reliable prediction of sensory properties by instrumental characterisation. It is therefore possible to propose the application of this sensory-instrumental tool to all the stakeholders involved in apple production and marketing, to have a reliable description of apple fruit quality.

Essays in Industrial Organization and Information Technology

Chapter 1 studies how consumers’ switching costs affect the pricing and profits of firms competing in two-sided markets such as Apple and Google in the smartphone market. When two-sided markets are dynamic – rather than merely static – I show that switching costs lower the first-period price if network externalities are strong, which is in contrast to what has been found in one-sided markets. By contrast, switching costs soften price competition in the initial period if network externalities are weak and consumers are more patient than the platforms. Moreover, an increase in switching costs on one side decreases the first-period price on the other side. Chapter 2 examines firms’ incentives to invest in local and flexible resources when demand is uncertain and correlated. I find that market power of the monopolist providing flexible resources distorts investment incentives, while competition mitigates them. The extent of improvement depends critically on demand correlation and the cost of capacity: under social optimum and monopoly, if the flexible resource is cheap, the relationship between investment and correlation is positive, and if it is costly, the relationship becomes negative; under duopoly, the relationship is positive. The analysis also sheds light on some policy discussions in markets such as cloud computing. Chapter 3 develops a theory of sequential investments in cybersecurity. The regulator can use safety standards and liability rules to increase security. I show that the joint use of an optimal standard and a full liability rule leads to underinvestment ex ante and overinvestment ex post. Instead, switching to a partial liability rule can correct the inefficiencies. This suggests that to improve security, the regulator should encourage not only firms, but also consumers to invest in security.

Many-Core Architectures: Hardware-Software Optimization and Modeling Techniques

During the last few decades an unprecedented technological growth has been at the center of the embedded systems design paramount, with Moore’s Law being the leading factor of this trend. Today in fact an ever increasing number of cores can be integrated on the same die, marking the transition from state-of-the-art multi-core chips to the new many-core design paradigm. Despite the extraordinarily high computing power, the complexity of many-core chips opens the door to several challenges. As a result of the increased silicon density of modern Systems-on-a-Chip (SoC), the design space exploration needed to find the best design has exploded and hardware designers are in fact facing the problem of a huge design space. Virtual Platforms have always been used to enable hardware-software co-design, but today they are facing with the huge complexity of both hardware and software systems. In this thesis two different research works on Virtual Platforms are presented: the first one is intended for the hardware developer, to easily allow complex cycle accurate simulations of many-core SoCs. The second work exploits the parallel computing power of off-the-shelf General Purpose Graphics Processing Units (GPGPUs), with the goal of an increased simulation speed. The term Virtualization can be used in the context of many-core systems not only to refer to the aforementioned hardware emulation tools (Virtual Platforms), but also for two other main purposes: 1) to help the programmer to achieve the maximum possible performance of an application, by hiding the complexity of the underlying hardware. 2) to efficiently exploit the high parallel hardware of many-core chips in environments with multiple active Virtual Machines. This thesis is focused on virtualization techniques with the goal to mitigate, and overtake when possible, some of the challenges introduced by the many-core design paradigm.