Fusarium head blight (FHB), caused mainly by Fusarium graminearum, is one of the most destructive global diseases of small cereal grains worldwide. The most devastating effect of this disease is the deposition of mycotoxins in the grain. Deoxynivalenol (DON) and its analogs 3-ADON, 15-ADON and NIV are the major mycotoxins produced by the fungus. The chemotype, 15-ADON is more prevalent in North America, but recently a chemotypic shift has been observed. The rapid emergence of the 3-ADON chemotype over the traditional 15-ADON chemotype is a major concern in North America. The studies in this thesis were focused on understanding the behaviour of the 3-ADON and 15-ADON chemotypes and the possible reasons for the rapid chemotypic shift.

Milk and dairy products are important source of bioactive compounds useful to satisfy the nutritional and physiological needs of any newborns of mammalian species and useful to guarantee adequate growth and development of infants as well as provide a complete nourishment of adults. Physico-chemical, nutritional and organoleptic properties of the main constituents and the “minor” components have a crucial role in the quality of milk and milk products. Although in the past decades dietary milk fat was often regarded as harmful for the human health, recent researches suggest that milk contains specific fatty acids with nutritional and physiological health benefits. For these reasons, a major attention is given to the quantity and quality of total fat intake. In the recent years, as a result of the new concept of multifunctional agriculture and the changing behaviours about diet, consumer demands in favor of high-quality, security and safety dairy products are increased. Moreover, milk proteins and milk-derived bioactive peptides are recognized to have a high nutritive value, several health-promoting functional activities and excellent technological properties. Accordingly, growing interest in the development of functional dairy products and preparation of infant formulae for babies who cannot be breast-fed, has been give in order to meet the specific consumer’s requests. This manuscript presents the main results obtained during my PhD research aimed to evaluate the main bioactive lipids and proteins in milk and dairy products using innovative analytical techniques. The experimental section of this manuscript is divided in two sections where are reported the main results obtained during my research activities on dairy products and human milks in order to characterize their bioactive compounds for functional food applications.

From the mid-fifteenth century onwards, the Scottish aristocratic community made increasing use of formal bonds of lordship, service and friendship. The first section of this thesis examines the relationship between formal bonding and the pursuit of feud and tenurial disputes. Written witnessed, bonds, particularly bonds of mutual friendship or defence, seem to have acquired a specific and, partly, symbolic role in the amicable arbitration of feud outwith formal courts of law. The bond was employed as a pledge for the good behaviour of previously hostile parties towards one another, guaranteeing the material terms of any settlement between them, and bolstering the newly-established state of non-aggression. Bonds used in this context were not primarily, designed to initiate long-term social and political cooperation between the contracting parties. The proliferation of bonds of friendship used in this way during the fifteenth century may perhaps be linked to the demands of royal courts for documentary evidence of amicable settlement. Bonds of maintenance, and bonds of manrent or retinue, were also used extensively in the settlement of feud, and in consolidating strained, or new, tenurial relationships. Bonds of service given in return for grants of lands, were often connected to attempts to keep the tenure of disputed lands highly conditional, and were typically linked to liferent and/or reversionary grants. The linking of tenure with formal bonds of service also occurred in areas, and periods, where the granter of land had cause to seek assurances of political loyalty and support which were more binding than the oaths and ceremonies associated with routine acts of feudal conveyancing. The general pattern suggests that, although all bonds of service appear to offer undefined open-ended service, the tenurial and political context in which these bonds were given did, on many occasions, define and limit the way in which maintenance or service was to be discharged. Bonds of all types were also used to obtain immediate political or military support in specific disputes. The remainder of the thesis deals with the interaction between local feud and 'national' politics. An analysis of the rebellion of Prince James (later James IV) against his father, James III, during 1488, indicates that many individual noblemen and prelates committed themselves to the rebellion in pursuit of local feuds and ambitions. After James III's death at Sauchieburn, the ascendancy of Prince James' supporters within their own localities was confirmed by individual acts of royal patronage and by parliamentary legislation, a process which generated more feuds. The behaviour of the new regime, and its persecution of men who had remained loyal to James III during 1488, resulted in a major rebellion during 1489. The rebellion was eventually ended by negotiation, and by the new regime making several important concession to rebel demands. Apparently incomprehensible changes of allegiance by major noblemen during the period 1487-9 can be shown to have been perfectly consistent in terms of the smaller disputes in which they were directly involved. An examination of the political career of James, Earl of Buchan suggests that violence remained a viable political tool for the fifteenth century nobility at both the local and national level, and, indeed, that the division between local and national politics was, in many cases, non-existent. The final three chapters exhibit the effect of changes in royal policy and patronage in generating violence and feud within the localities, and the part this could play in provoking direct opposition to the crown. The importance of these tensions may have grown during the course of the fifteenth century as the amount of land, and the number of offices, under direct royal control grew through the forfeiture of several major landowning families and the annexation of their estates to the royal patrimony.

The existence of Multiple Myeloma Stem cells (MMSCs)is supposed to be one of the major causes of MM drug-resistance. However, very little is known about the molecular characteristics of MMSCs, even if some studies suggested that these cells resembles the memory B cells. In order to molecularly characterize MMSCs, we isolated the 138+138- population. For each cell fraction we performed a VDJ rearrangement analysis. The complete set of aberrations were performed by SNP Array 6.0 and HG-U133 Plus 2.0 microarray analyses (Affymetrix). The VDJ rearrangement analyses confirmed the clonal relationship between the 138+ clone and the immature clone. Both BM and PBL 138+ clones showed exactly the same genomic macroalterations. In the BM and PBL 138-19+27+ cell fractions several micro-alterations (range: 1-350 Kb) unique of the memory B cells clone were highlighted. Any micro-alterations detected were located out of any genomic variants region and are presumably associated to the MM pathogenesis, as confirmed by the presence of KRAS, WWOX and XIAP genes among the amplified regions. To get insight into the biology of the clonotypic B cell population, we compared the gene expression profile of 8 MM B cells samples 5 donor B cells vs, thus showing a differential expression of 11480 probes (p-value: <0,05). Among the self-renewal mechanisms, we observed the down-regulation of Hedgehog pathway and the iperactivation of Notch and Wnt signaling. Moreover, these immature cells showed a particular phenotype correlated to resistance to proteasome inhibitors (IRE1α-XBP1: -18.0; -19.96. P<0,05). Data suggested that the MM 138+ clone might resume the end of the complex process of myelomagenesis, whereas the memory B cells have some intriguing micro-alterations and a specific transcriptional program, supporting the idea that these post germinal center cells might be involved in the transforming event that originate and sustain the neoplastic clone.

The human p53 tumor suppressor, known as the “guardian of the genome”, is one of the most important molecules in human cancers. One mechanism for suppressing p53 uses its negative regulator, MDM2, which modulates p53 by binding directly to and decreasing p53 stability. In testing novel therapeutic approaches activating p53, we investigated the preclinical activity of the MDM2 antagonist, Nutlin-3a, in Philadelphia positive (Ph+) and negative (Ph-) leukemic cell line models, and primary B-Acute lymphoblastic leukemia (ALL) patient samples. In this study we demonstrated that treatment with Nutlin-3a induced grow arrest and apoptosis mediated by p53 pathway in ALL cells with wild-type p53, in time and dose-dependent manner. Consequently, MDM2 inhibitor caused an increase of pro-apoptotic proteins and key regulators of cell cycle arrest. The dose-dependent reduction in cell viability was confirmed in primary blast cells from Ph+ ALL patients with the T315I Bcr-Abl kinase domain mutation. In order to better elucidate the implications of p53 activation and to identify biomarkers of clinical activity, gene expression profiling analysis in sensitive cell lines was performed. A total of 621 genes were differentially expressed (p < 0.05). We found a strong down-regulation of GAS41 (growth-arrest specific 1 gene) and BMI1 (a polycomb ring-finger oncogene) (fold-change -1.35 and -1.11, respectively; p-value 0.02 and 0.03, respectively) after in vitro treatment as compared to control cells. Both genes are repressors of INK4/ARF and p21. Given the importance of BMI in the control of apoptosis, we investigated its pattern in treated and untreated cells, confirming a marked decrease after exposure to MDM2 inhibitor in ALL cells. Noteworthy, the BMI-1 levels remained constant in resistant cells. Therefore, BMI-1 may be used as a biomarker of response. Our findings provide a strong rational for further clinical investigation of Nutlin-3a in Ph+ and Ph-ALL.

In chronic myeloid leukemia and Philadelphia-positive acute lymphoblastic leukemia patients resistant to tyrosine kinase inhibitors (TKIs), BCR-ABL kinase domain mutation status is an essential component of the therapeutic decision algorithm. The recent development of Ultra-Deep Sequencing approach (UDS) has opened the way to a more accurate characterization of the mutant clones surviving TKIs conjugating assay sensitivity and throughput. We decided to set-up and validated an UDS-based for BCR-ABL KD mutation screening in order to i) resolve qualitatively and quantitatively the complexity and the clonal structure of mutated populations surviving TKIs, ii) study the dynamic of expansion of mutated clones in relation to TKIs therapy, iii) assess whether UDS may allow more sensitive detection of emerging clones, harboring critical 2GTKIs-resistant mutations predicting for an impending relapse, earlier than SS. UDS was performed on a Roche GS Junior instrument, according to an amplicon sequencing design and protocol set up and validated in the framework of the IRON-II (Interlaboratory Robustness of Next-Generation Sequencing) International consortium.Samples from CML and Ph+ ALL patients who had developed resistance to one or multiple TKIs and collected at regular time-points during treatment were selected for this study. Our results indicate the technical feasibility, accuracy and robustness of our UDS-based BCR-ABL KD mutation screening approach. UDS was found to provide a more accurate picture of BCR-ABL KD mutation status, both in terms of presence/absence of mutations and in terms of clonal complexity and showed that BCR-ABL KD mutations detected by SS are only the “tip of iceberg”. In addition UDS may reliably pick 2GTKIs-resistant mutations earlier than SS in a significantly greater proportion of patients.The enhanced sensitivity as well as the possibility to identify low level mutations point the UDS-based approach as an ideal alternative to conventional sequencing for BCR-ABL KD mutation screening in TKIs-resistant Ph+ leukemia patients

Backgrounds:Treatment of patients with relapsed/refractory (R/R) diffuse large B-cell lymphoma (DLBCL) not eligible to high dose therapy represents an unmet medical need. Panobinostat showed encouraging therapeutic activity in studies conducted in lymphoma cell lines and in vivo in patients with advanced hematologic malignancies.Purpose:FIL-PanAL10 (NCT01523834) is a phase II, prospective multicenter trial of the Fondazione Italiana Linfomi (FIL) to evaluate safety and efficacy of single agent Panobinostat as salvage therapy for R/R DLBCL patients and to evaluate a possible relationships between response and any biological features. Patients and Methods:Patients with R/R DLBCL were included. The treatment plan included 6 induction courses with Panobinostat monotherapy followed by other 6 courses of consolidation. The primary objective was to evaluate Panobinostat activity in terms of overall response (OR); secondary objectives were: CR rate, time to response (TTR), progression-free survival (PFS), safety and feasibility of Panobinostat. We included evaluation of the impact of pharmacogenetics, immunohistochemical patterns and patient’s specific gene expression and mutations as potential predictors of response to Panobinostat as explorative objectives. To this aim a pre-enrollment new tissue biopsy was mandatory. ResultsThirty-five patients, 21 males (60%), were enrolled between June 2011 and March 2014. At the end of induction phase, 7 responses (20%) were observed, including 4 CR (11%), while 28 patients (80%) discontinued treatment due to progressive disease (PD) in 21 (60%) or adverse events in 7 (20%). Median TTR in 9 responders was 2.6 months (range 1.8-12). With a median follow up of 6 months (range 1-34), the estimated 12 months PFS and OS were 27% and 30.5%, respectively. Grade 3-4 thrombocytopenia and neutropenia were the most common toxicities (in 29 (83%) and 12 (34%) patients, respectively. Conclusions The results of this study indicate that Panobinostat might be remarkably active in some patients with R/R DLBCL, showing durable CR

L’enzima IDO interviene nella via di degradazione del triptofano, essenziale per la vita cellulare; l’iperespressione di IDO favorisce la creazione di un microambiente immunotollerante. Nelle LAM IDO è funzionalmente attivo nelle cellule blastiche e determina l’acquisizione di un fenotipo regolatorio da parte delle cellule T alloreattive; l’espressione della proteina aumenta in modo consensuale con l’evoluzione clinica della patologia. Scopo della Tesi è indagare l’esistenza di una correlazione tra l’iperespressione di IDO da parte delle cellule leucemiche, le caratteristiche di rischio alla diagnosi e l’outcome dei pazienti. Sono stati esaminati 45 pazienti adulti affetti da LAM afferiti all’Istituto di Ematologia di Bologna. I pazienti sono stati stratificati a seconda di: età di insorgenza della leucemia, secondarietà a Mielodisplasia o radio chemioterapia, iperleucocitosi, citogenetica, biologia molecolare (sono state valutate le alterazioni a carico dei geni FLT3 ed NPM). I pazienti sono stati analizzati per l’espressione del gene IDO mediante RT-PCR, seguita da Western Blot, allo scopo di stabilire la presenza di una proteina attiva; successivamente si è proceduto a verificare l’esistenza di una correlazione tra l’espressione di IDO e le caratteristiche di rischio alla diagnosi per identificare una relazione tra l’espressione del gene ed un subset di pazienti a prognosi favorevole o sfavorevole. Dei 45 pazienti adulti affetti da LAM il 28,9% è risultato negativo per l’espressione di IDO, mentre il rimanente 71,1% è risultato positivo ed è stato suddiviso in tre ulteriori categorie, in base ai livelli di espressione. I dati non sembrano al momento suggerire l’esistenza di una correlazione tra l’espressione di IDO e le caratteristiche di rischio alla diagnosi. Nel gruppo di pazienti ad elevata espressione di IDO si riscontra un rate di resistenza alla chemioterapia di induzione più elevato, con una quota di pazienti resistenti pari al 71,4%, contro il 23,1% nel gruppo di pazienti IDO-negativi. / Tryptophan is an essential amino-acid required for protein synthesis and for cell proliferation. IDO is an eme-containing enzyme that catalyzes the rate-limiting step in tryptophan degradation along the kynurenine pathway and its overexpression is known to be involved in immune tolerance induction. IDO is functionally active in Acute Myeloid Leukemia tumor cells and it acts by inducing the transformation of T-cells into regulatory T-cells. Several observations demonstrated that IDO expression increases along with disease progression. The object of the work is to establish a correlation between IDO expression by leukemic cells, risk factors at diagnosis and patients’ outcome. Adult AML patients from the Hematology Institute “L. and A. Seràgnoli” in Bologna were analyzed for risk characteristics at diagnosis and for IDO expression. Patients were stratified according to: age at diagnosis, de novo or secondary disease (pre-existing myelodysplastic syndrome or radio-chemotherapy), hyperleucocytosis, cytogenetics (on the basis of cytogenetic characteristics patients were divided into low, intermediate or high risk group), molecular biology (the presence of FLT3 and NPM alterations was studied). These patients were analyzed for IDO expression by RT-PCR and the presence of a functionally active IDO protein was established by Western Blotting analysis. Eventually the correlation between IDO expression, risk factors at diagnosis and patients’ outcome was evaluated. Thirteen out of the 45 analyzed patients (28,9%) resulted to be IDO-negative, 71,1% being IDO-positive. Positive patients were divided into three different subgroups according to the IDO level. No significant differences in the recurrence of prognostic characteristics at diagnosis between IDO-negative and IDO-positive patients were experienced. Intriguingly, it has been found that refractory patients were 71,4% among patients who express IDO at high level and 23,1% among IDO-negative patients.

Thesis (Ph. D.)--University of Oregon, 2008. / Typescript. Includes vita and abstract. Includes bibliographical references (leaves 69-81). Also available online in ProQuest, free to University of Oregon users.

Bakgrund och problem: Den 1 januari 2018 trädde den nya intäktsstandarden IFRS 15 Intäkter från avtal med kunder i kraft. Alla företag som redovisar enligt IFRS och är verksamma inom EU måste därmed anpassa sin redovisning efter den. Standarden anses komplex då den innebär ett nytt synsätt på intäktsredovisning. Företagen ställs inför förändringar och implementeringsprocesser, som för de flesta utgör svårigheter och problem. De mest berörda branscherna är bygg och entreprenad, IT och telekom samt detaljhandeln. Syfte: Studien syftar till att undersöka hur företagen har implementerat IFRS 15. Genom det önskar vi öka kunskapen om vilka faktorer som har betydande påverkan vid implementering av en standard. Metod: Studien har en kvalitativ forskningsmetod. Företag från de mest berörda branscherna har valts ut. För att säkerställa att företagen kan bidra till studien, har ett urval gjorts via analys av bokslutskommunikeer. Företag som uttryckt att IFRS 15 kommer ha väsentlig påverkan har kontaktats. Enkätfrågor rörande planering, utbildning, målformulering samt resultat har sedan besvarats av tre företag inom branscherna bygg och entreprenad samt IT och Telekom. Slutsatser: Samtliga företag har via en välplanerad handlingsplan, med fokus på tidsplanering, haft ett proaktivt arbetssätt. Korta och koncisa mål, internutbildningar samt val av utbildningsmetod har haft påverkan på implementeringen. Engagemang från chefer och ledning i form av aktivt deltagande har prioriterats i två av företagen, men haft lägre prioritet hos det tredje. Användning av externhjälp i form av konsulter och projektledare har ökat kunskapsnivån hos företagen. Sammanfattningsvis har tre betydande faktorer vid implementeringen av IFRS 15 identifierats: mål och handlingsplan, utbildning och kompetens samt chefers och ledningens engagemang. / Background and problem: January 1st, 2018, the new revenue standard IFRS 15 - Revenue from contracts with customers became effective. All companies that report in accordance with IFRS and are domiciled in the EU must therefore adjust their reporting accordingly. The standard is considered complex as it contributes to a new approach to revenue recognition. Companies face changes and implementation processes that generally contribute to difficulties and problems. The most affected industries are building and construction, IT and telecommunications and retail. Purpose: The study aims at investigating how companies have implemented IFRS 15. By this, we wish to increase knowledge about which factors that have significant impact on implementing a standard. Method: The study has a qualitative research method. Companies from the most affected industries have been selected. In order to ensure that the selected companies can contribute to the study, disclosures of changes in accounting policies, in the interim reports for the fourth quarter of 2017, were analyzed. Companies that expressed that IFRS 15 will have a significant impact has been contacted. Survey questions regarding planning, education, goal formulation and results have been answered by three companies in the building and construction industries as well as IT and Telecom. Conclusion: Through a well-planned action plan, focusing on time management, all three companies have had a proactive approach. Short and concise goals, internal training and choice of education methods have had an impact on the implementation. Commitment from managers and management by active participation has been prioritized in two of the companies, but had a lower priority in the third. The use of external assistance in the form of consultants and project managers has increased the level of knowledge in the companies. In summary, three significant factors in the implementation of IFRS 15 have been identified: goals and action plan, education and competence, and the commitment of managers and management.