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The enantioselective reduction of ketoimines has been successfully realized, using trichlorosilane as the stoichiometric reducing agent in the presence of catalytic amounts of a Lewis base, specifically a Cinchona derivative. For the first time, a novel class of derivatives was studied, featuring a picolinamide unit bound to the alkaloid scaffold, further functionalized as quaternary ammonium salt at the quinuclidine ring. Excellent yields and from good to high enantioselectivities (up to 92% ee) were obtained in the reduction of ketoimines. The novel catalysts were successfully employed in the synthesis of an enantiomerically pure advanced precursor of the blockbuster drug Rivastigmine.

Adult T cell Leukemia/Lymphoma (ATLL) is a mature T cell malignancy associated with Human T cell Leukemia Virus type 1 (HTLV-1) infection. Among its four main clinical subtypes, the prognosis of acute and lymphoma variants remains poor. The long latency (3–6 decades) and low incidence (3–5%) of ATLL imply the involvement of viral and host factors in full-blown malignancy. Despite multiple preclinical and clinical studies, the contribution of the stromal microenvironment in ATLL development is not yet completely unraveled. The aims of this study were to investigate the role of the host microenvironment, and specifically fibroblasts, in ATLL pathogenesis and to propose a murine model for the lymphoma subtype. Here we present evidence that the oncogenic capacity of HTLV-1-immortalized C91/PL cells is enhanced when they are xenotransplanted together with human foreskin fibroblasts (HFF) in immunocompromised BALB/c Rag2-/-γc-/- mice. Moreover, cell lines derived from a developed lymphoma and their subsequent in vivo passages acquired the stable property to induce aggressive T cell lymphomas. In particular, one of these cell lines, C91/III cells, consistently induced aggressive lymphomas also in NOD/SCID/IL2Rγc KO (NSG) mice. To dissect the mechanisms linked to this enhanced tumorigenic ability, we quantified 45 soluble factors released by these cell lines and found that 21 of them, mainly pro-inflammatory cytokines and chemokines, were significantly increased in C91/III cells compared to the parental C91/PL cells. Moreover, many of the increased factors were also released by human fibroblasts and belonged to the known secretory pattern of ATLL cells. C91/PL cells co-cultured with HFF showed features reminiscent of those observed in C91/III cells, including a similar secretory pattern and a more aggressive behavior in vivo. On the whole, our data provide evidence that fibroblasts, one of the major stromal components, might enhance tumorigenesis of HTLV-1-infected and immortalized T cells, thus throwing light on the role of microenvironment contribution in ATLL pathogenesis. We also propose that the lymphoma induced in NSG mice by injection with C91/III cells represents a new murine preclinical ATLL model that could be adopted to test novel therapeutic interventions for the aggressive lymphoma subtype.

PES (2-phenylethynesulfonamide) was initially identified as an inhibitor of p53 translocation to mitochondria and named Pifithrin-µ. Further studies showed that PES selectively killed tumour cells and was thus a promising anticancer agent. PES-induced cell death was characterised by a non-apoptotic, autophagosome-rich phenotype. We observed this phenotype via electron microscopy in wild type (wt) and double Bax-/- Bak-/- (DKO) mouse embryonic fibroblasts (MEFs) treated with PES. We excluded the involvement of effector caspases, BAX and BAK, in causing PES-triggered cell death. Therefore, apoptosis was ruled out as the lethal mode of action of PES. Surprisingly, MEFs containing BAX were significantly protected from PES treatments. BAX overexpression in Bax-/- MEFs confirmed this pro-survival effect. Moreover, this protective effect required the ability of BAX to localise to mitochondrial membranes. Conversely, mitochondrial fusion induced by treatment with Mdivi-1 conferred increased resistance to MEFs subjected to PES treatment. The involvement of BAX in the regulation of mitochondrial dynamics has been reported. We propose the promotion of mitochondrial fusion by BAX to be the pro-survival function attributed to BAX. This work was supported by the Spanish Government, MINECO (Ministerio de Economía y Competitividad), project SAF2011-29730. Paolo Mattiolo was supported by an FI fellowship from AGAUR (Generalitat de Catalunya).

2-Phenylethynesulfonamide (PES) or pifithrin-μ is a promising anticancer agent with preferential toxicity for cancer cells. The type of cell death and the molecular cascades activated by this compound are controversial. Here, we demonstrate PES elicits a caspase- and BAX/BAK-independent non-necroptotic necrotic cell death, since it is not inhibited by necrostatin-1. This process is characterized by an early generation of reactive oxygen species (ROS) resulting in p53 up-regulation. Accordingly, thiolic antioxidants protect cells from PES-induced death. Furthermore, inhibiting the natural sources of glutathione with l-buthionine-sulfoximine (BSO) strongly cooperates with PES in triggering cytotoxicity. Genetically modified p53-null or p53 knocked-down cells show resistance to PES-driven necrosis. The predominant localization of p53 in chromatin-enriched fractions added to the up-regulation of the p53-responsive gene p21, strongly suggest the involvement of a transcription-dependent p53 program. On the other hand, we report an augmented production of ROS in p53-positive cells that, added to the increased p53 content in response to PES-elicited ROS, suggests that p53 and ROS are mutually regulated in response to PES. In sum, p53 up-regulation by ROS triggers a positive feedback loop responsible of further increasing ROS production and reinforcing PES-driven non-necroptotic necrosis.

Many potentially modifiable risk factors for MS are investigated. It is not known, however, if these factors also apply to MS-related cognitive impairment (CI), a frequent consequence of MS.The aim of our study was to assess risk factors for CI in MS patients, focusing on environmental exposures, lifestyle and comorbidities.We included MS patients referring to MS Centers in Florence and Barletta between 2014 and 2017. Neuropsychological performance was assessed through the Rao's battery and Stroop test, cognitive reserve (premorbid intelligence quotient-IQ) was evaluated using the National Adult Reading Test (NART). Potential risk factors were investigated through a semi-structured questionnaire.150 patients were included. CI was detected in 45 (30%) subjects and was associated with older age (p0.005), older age at MS onset (p = 0.016), higher EDSS score (p0.005), progressive disease course (p = 0.048) and lower premorbid IQ score (p0.005). As for risk factors, CI was related with lower physical activity in childhood-adolescence (p0.005). In women, hormonal therapy resulted to be protective against CI (p = 0.041). However, in the multivariable analysis, the only significant predictors of CI were older age (p0.05; OR 1.06, 95% CI 1.02-1.10) and lower premorbid IQ (p0.05; OR 0.93, 95% CI: 0.88-0.98). Removing IQ from the model, CI was associated with higher EDSS (p = 0.030; OR 1.25, 95% CI 1.02-1.53) and, marginally, previous physical activity (p = 0.066; OR 0.49, 95% CI: 0.23-1.05).Our findings suggest that physical activity in childhood-adolescence could be a contributor to cognitive reserve building, thus representing a potential protective factors for MS-related CI susceptible to preventive strategies.

In generale, il 2015 è stato un anno decisamente diversificato per quanto riguarda le produzioni, ma certamente più favorevole per la coltura dell’orzo primaverile al Centro-Nord, che dopo due anni negativi si riallinea al 2012 con una media produttiva di 5,3 t/ha.

La rete delle prove nazionali di orzo da zootecnia per l’annata 2014-2015 mostra risultati molto diversi a seconda della località. La media produttiva al Nord risulta di 6,5 t/ha come lo scorso anno, al Centro in deciso aumento con 6,7 t/ha, mentre al Sud la produttività evidenzia una perdita di 1 t/ha.

This work aimed at developing a HS–SPME/GC–MS method for the extraction and analysis of volatile compounds from the kernels of durum wheat. Fiber coating, temperature and time of extraction were evaluated. Moreover, the method was used as a tool for varietal characterization. The qualitative and semi-quantitative characterization of the volatile fraction of durum wheat kernels highlighted 11 different chemical classes, alcohols and aldehydes prevailing over acids, alkanes, aromatic and heterocyclic hydrocarbons. Concerning the comparison among the studied durum wheat kernel cultivars, Sculptur was the one with the highest amount and variability of volatile compounds. This cultivar stood out for the highest level of alkanes and ketones, while Anco Marzio contained higher levels of aldehydes and alkenes. The optimized method allowed for the qualitative and semi-quantitative characterization of the volatile fraction of durum wheat kernels, and it was effective at differentiating durum wheat cultivars based on their different volatile profiles, representing a potential tool for varietal selection and for the exploitation of specific durum wheat cultivars.

Al Nord la media dell' areale è stata di 6,5 t/ha in ripresa rispetto al tracollo della scorsa annata, al Centro è risultata di sole 5,1 t/ha, dato al di sotto delle potenzialità produttive; al Sud la produttività media è stata di 5,7 t/ha, un ottimo risultato che riporta l' areale su livelli elevati

La rete di prove di confronto varietale di grano duro, condotta in Siclia con il coordinamento del CRA-ACM, ha previsto nell’annata 2013-2014 l’allestimento di 4 campi in differenti ambienti: Libertinia (Catania), Catania, Cammarata e S. Stefano Quisquina (Agrigento). Fra le sette varietà al primo anno di valutazione, emergono le performance di Antalis, che ha manifestato ottima capacità produttiva, precocita’ di spigatura, buone caratteristiche merceologiche, contenuto proteico medio e buona resistenza alle malattie.

Per il Centro-sud e Isole la produzione ha superato di 0,3 t/ha quella dell’annata precedente attestandosi a 4,8 t/ha, mentre per il Centro-nord la media è stata di sole 3,1 t/ha. La sofferenza della coltura si è manifestata chiaramente anche in termini di ritardo della spigatura, avvenuta nell’ultima decade di maggio

We present here a case of immune reconstitution inflammatory syndrome associated with Kaposi’s sarcoma (KS-IRIS) developed in an AIDS patient two months after initiation of antiretroviral therapy (ART). Baseline characteristics of this IRIS-KS case, within a cohort of 12 naïve AIDS-KS patients, were analyzed. No statistically significant differences in CD4 cell counts, plasma HIV RNA load, KS clinical staging, human herpesvirus 8 (HHV8) antibody titers and HHV8 load in peripheral blood mononuclear cells and saliva were evidenced. HHV8 load in plasma was found to be significantly higher in the KS-IRIS patient (> 6 log10 genome equivalents/ml, p = 0.01, t–test) compared to the 11 patients with KS regression. This case highlights that measurement of HHV8 load in plasma may be useful to identify patients at risk for KS-IRIS, and that this parameter should be included in the design of larger studies to define KS-IRIS risk predictors.

The 1st part of the thesis demonstrates that autophagy promotes cell death by an intrinsic apoptotic pathway at the early times of trophic-nutritional deprivation. In this paradigm, autophagy also promotes apoptosis induced by anticancer drugs. At longer times, autophagy displays its assumed protective role on starved cells. Time, being the only determinant of the dual role of autophagy on cell fate, is a novelty that impinges on tumor biology and therapy. The 2nd part of the thesis focuses on the compound 2-phenylethynesulfonamide (PES) with a promising lethal selectivity for cancer cells, already published. PES is an inhibitor of Heat Shock Protein 70 and autophagy, thus impairing cell proteostasis. The thesis reveals a positive feed-back between reactive oxygen species and p53 in the PES mode of action. Finally, PES uncovers an unexpected pro-survival function of BAX protein, precisely the promotion of mitochondrial fusion., La primera part de la tesi demostra que l'autofàgia promou la mort cel•lular per una via apoptòtica intrínseca a temps inicials de la privació-tròfica nutricional. En aquest paradigma, l'autofàgia també promou l’apoptosi induïda per fàrmacs anticancerosos. A temps més llargs, l'autofàgia mostra el seu paper protector. El temps com a únic determinant de la funció dual de l'autofàgia en el destí cel•lular és una novetat, que incideix en la biologia i teràpia antitumoral. La segona part de la tesi estudia el compost 2-feniletísulfonamida (PES) amb una prometedora letalitat selectivitat pel càncer, ja publicada. PES és un inhibidor de “Heat Shock Protein 70” i autofàgia, fet que trenca la proteostasi cel•lular. La tesi revela una retroalimentació positiva entre espècies reactives d’oxigen i p53 en el mecanisme d’acció de PES. Finalment, PES posa de manifest una funció inesperada i pro-supervivència de la proteïna BAX, concretament promoure la fusió mitocondrial., La primera parte de la tesis demuestra que la autofagia promueve la muerte celular por vía apoptótica intrínseca a tiempos tempranos de privación trófico-nutricional. En este paradigma, la autofagia también promueve apoptosis inducida por fármacos anticancerosos. A tiempos más largos, la autofagia muestra su papel protector. El tiempo como único determinante de la función dual de la autofagia en el destino celular es una novedad, que incide en la biología y terapia antitumoral. La segunda parte de la tesis estudia el compuesto 2-feniletinosulfonamida (PES) con una prometedora letalidad selectiva por el cáncer, ya publicada. PES es un inhibidor de "Heat Shock Protein 70" y autofagia, lo cual rompe la proteostasis celular. La tesis revela una retroalimentación positiva entre especies reactivas de oxígeno y p53 en el mecanismo de acción de PES. Finalmente, PES pone de manifiesto una función inesperada y pro-supervivencia de la proteína BAX, concretamente promover la fusión mitocondrial.

Autophagy is generally regarded as a mechanism to promote cell survival. However, autophagy can occasionally be the mechanism responsible of cell demise. We have found that a concomitant depletion of glucose, nutrients and growth factors provoked cell death in a variety of cell lines. This death process was contingent upon caspase activation and was mediated by BAX/BAK proteins, thus indicating its apoptotic nature and the engagement of an intrinsic pathway. In order to abrogate autophagy, 3-methyladenine (3-MA), BECLIN-1 siRNA and Atg5 knock-out (Tet-Off type) approaches were alternatively employed. Irrespective of the procedure, at short times of starvation, we found that the ongoing autophagy was sensitizing cells to the permeabilization of the mitochondrial outer membrane (MOMP), caspase activation and, therefore, apoptosis. On the contrary, at longer times of starvation, autophagy displayed its characteristic pro-survival effect on cells. As far as we know, we provide the first experimental paradigm where time is the only variable determining the final outcome of autophagy. In other words, we have circumscribed in time the shift transforming autophagy from a cell death to a protection mechanism. Moreover, at short times, starvation-driven autophagy exacerbated the apoptotic cell death caused by several antitumor agents. In agreement with this fact, their apoptotic effects were greatly diminished by autophagy inhibition. The implications of these facts in tumor biology will be discussed. The author’s thanks Dr. P. Codogno for the Atg5 Tet-Off MEFs m5-7 (originated at Prof. Mizushima’s laboratory) and Dr. S.E. Lupold for the PC3 and DU145 cell lines. This work was supported by Ministerio de Economía y Competitividad SAF2011-29730. VJY is under a “Retention of Research Talent” contract of “Programa Banco de Santander”. PM was supported by a fellowship (AH9815758) from AGAUR (Generalitat de Catalunya). University of Lleida Funds (Ajuts Pont 2015) contributed to the completion of this work.

RNA interference (RNAi) has been suggested to represent a promising therapeutic approach in different disease settings. Primary effusion lymphoma (PEL) is a plasmablastic lymphoma consistently expressing B lymphocyte-induced maturation protein 1 (Blimp-1), a pivotal transcriptional regulator during

La conservazione della biodiversità di specie diverse ed in particolare del frumento duro è tra gli obiettivi prioritari della ricerca agronomica degli ultimi anni. Si tenta, infatti, di utilizzare le varietà tradizionalmente coltivate per assicurare il contenimento dell erosione genetica causata dall introduzione di nuove varietà ad alto livello produttivo, ma anche per reperire geni utilizzabili nel lavoro di miglioramento genetico per la valorizzazione delle materie prime per prodotti di nicchia. La caratterizzazione oggetto di questa ricerca, condotta durante il corso di dottorato in Produttività delle piante coltivate in ambiente Mediterraneo finanziata dal MIUR progetto PON ISCOCEM, è stata effettuata a partire dall esame elettroforetico dei genotipi al momento del reperimento (2003-04). Allo scopo sono stati analizzati caratteri agronomici, morfologici e qualitativi di popolazioni di frumento duro tradizionalmente coltivate in Sicilia. Sono state rilevate le principali fenofasi del ciclo biologico mentre i rilievi morfologici hanno interessato caratteri del culmo, della spiga e del seme. In particolare, per ciò che concerne quest ultimo aspetto, l esame dei caratteri morfologici rilevati ha messo in luce differenze a volte marcate e a volta di scarsa entità rispetto alla caratterizzazione rilevata in letteratura da attribuire alle condizioni di maturazioni in un ambiente particolarmente stressante come la Piana di Catania (es. colore delle reste) o ai maggiori input agronomici (es. forma della spiga). Tra i caratteri qualitativi sono stati analizzati il contenuto proteico, il contenuto in fibra solubile (beta-glucano) nota per le azioni benefiche sulla salute dell uomo. La differenziazione dei genotipi è stata condotta anche attraverso lo studio dell amido effettuando un rilievo fotografico dei granuli per mezzo di microscopio a scansione elettronica (SEM) e la valutazione delle rispettive componenti quali amilosio e amilopectina. Le immagini al SEM hanno messo in rilievo la prevalenza di granuli di tipo A nelle varietà con maggiore precocità e la presenza di granuli più piccoli nelle varietà più tardive. Lo studio delle sostanze volatili, previa messa a punto del metodo, ha messo in luce differenze tra le popolazioni esaminate ma anche variazioni nei composti rilevati per effetto delle condizioni ambientali. Il regime idrico ha influenzato l espressione del genotipo mettendo sottolineando la presenza di alcuni composti come 3-metile-butanale solo in condizioni di stress idrico. Anche il regime termico sfavorevole ha determinato la sintesi produzione di sostanze volatili in quantità e qualità diverse a seconda dell ambiente di coltivazione come dimostrato per la cultivar Timilia. La ricerca può considerarsi il primo passo verso l individuazione di marker varietali e di qualità.

Aerobic metabolism of mammalian cells leads to the generation of reactive oxygen species (ROS). To cope with this toxicity, evolution provided cells with effective antioxidant systems like glutathione. Current anticancer therapies focus on the cancer dependence on oncogenes and non-oncogenes. Tumors trigger mechanisms to circumvent the oncogenic stress and to escape cell death. In this context we have studied 2-phenylethinesulfoxamine (PES), which disables the cell protective mechanisms to confront the proteotoxicity of damaged and unfolded proteins. Proteotoxic stress is increased in tumor cells, thus providing an explanation for the anticancer selectivity of PES. In addition, we have found that PES induces a severe oxidative stress and the activation of p53. The reduction of the cell content in glutathione by means of L-buthionine-sulfoximine (BSO) synergizes with PES. In conclusion, we have found that ROS constitutes a central element in a series of positive feed-back loops in the cell. ROS, p53, proteotoxicity, autophagy and mitochondrial dynamics are interconnected with the mechanisms leading to cell death, either apoptotic or necrotic. This network of interactions provides multiple targets for drug discovery and development in cancer.

The enantioselective redn. of ketoimines has been successfully realized, using trichlorosilane as the stoichiometric reducing agent in the presence of catalytic amts. of a Lewis base, specifically a Cinchona deriv. For the first time, a novel class of derivs. was studied, featuring a picolinamide unit bound to the alkaloid scaffold, further functionalized as quaternary ammonium salt at the quinuclidine ring. Excellent yields and from good to high enantioselectivities (up to 92% ee) were obtained in the redn. of ketoimines. The novel catalysts were successfully employed in the synthesis of an enantiomerically pure advanced precursor of the blockbuster drug rivastigmine.

Irradiation is a preservation technology used to improve the safety and hygienic quality of food. Aim of this study was to assess the applicability and validity of the microbiological screening method direct epifluorescence filter technique (DEFT)/aerobic plate count (APC) (EN 13783:2001) for the identification of irradiated herbs and spices. Tests on non-irradiated and irradiated samples of dried herbs and spices were performed. The method was based on the comparison of APC and count obtained using DEFT. In accordance with the standard reference, this method is not applicable to samples with APC3 colony forming units (CFU)/g and this is its main limit. The results obtained in our laboratories showed that in 50% of cases of non-irradiated samples and in 96% of the samples treated with ionising radiation, the method was not applicable due to a value of CFU/g3.