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UnlabelledAlthough HAART suppresses HIV replication, it is often unable to restore immune homeostasis. Consequently, non-AIDS-defining diseases are increasingly seen in treated individuals. This is attributed to persistent virus expression in reservoirs and to cell activation. Of note, in CD4(+) T cells and monocyte-macrophages of virologically-suppressed individuals, there is continued expression of multi-spliced transcripts encoding HIV regulatory proteins. Among them, Tat is essential for virus gene expression and replication, either in primary infection or for virus reactivation during HAART, when Tat is expressed, released extracellularly and exerts, on both the virus and the immune system, effects that contribute to disease maintenance. Here we report results of an ad hoc exploratory interim analysis (up to 48 weeks) on 87 virologically-suppressed HAART-treated individuals enrolled in a phase II randomized open-label multicentric clinical trial of therapeutic immunization with Tat (ISS T-002). Eighty-eight virologically-suppressed HAART-treated individuals, enrolled in a parallel prospective observational study at the same sites (ISS OBS T-002), served for intergroup comparison. Immunization with Tat was safe, induced durable immune responses, and modified the pattern of CD4(+) and CD8(+) cellular activation (CD38 and HLA-DR) together with reduction of biochemical activation markers and persistent increases of regulatory T cells. This was accompanied by a progressive increment of CD4(+) T cells and B cells with reduction of CD8(+) T cells and NK cells, which were independent from the type of antiretroviral regimen. Increase in central and effector memory and reduction in terminally-differentiated effector memory CD4(+) and CD8(+) T cells were accompanied by increases of CD4(+) and CD8(+) T cell responses against Env and recall antigens. Of note, more immune-compromised individuals experienced greater therapeutic effects. In contrast, these changes were opposite, absent or partial in the OBS population. These findings support the use of Tat immunization to intensify HAART efficacy and to restore immune homeostasis.Trial registrationClinicalTrials.gov NCT00751595.

Use of Env in HIV vaccine development has been disappointing. Here we show that, in the presence of a biologically active Tat subunit vaccine, a trimeric Env protein prevents in monkeys virus spread from the portal of entry to regional lymph nodes. This appears to be due to specific interactions between Tat and Env spikes that form a novel virus entry complex favoring R5 or X4 virus entry and productive infection of dendritic cells (DCs) via an integrin-mediated pathway. These Tat effects do not require Tat-transactivation activity and are blocked by anti-integrin antibodies (Abs). Productive DC infection promoted by Tat is associated with a highly efficient virus transmission to T cells. In the Tat/Env complex the cysteine-rich region of Tat engages the Env V3 loop, whereas the Tat RGD sequence remains free and directs the virus to integrins present on DCs. V2 loop deletion, which unshields the CCR5 binding region of Env, increases Tat/Env complex stability. Of note, binding of Tat to Env abolishes neutralization of Env entry or infection of DCs by anti-HIV sera lacking anti-Tat Abs, which are seldom present in natural infection. This is reversed, and neutralization further enhanced, by HIV sera containing anti-Tat Abs such as those from asymptomatic or Tat-vaccinated patients, or by sera from the Tat/Env vaccinated monkeys. Thus, both anti-Tat and anti-Env Abs are required for efficient HIV neutralization. These data suggest that the Tat/Env interaction increases HIV acquisition and spreading, as a mechanism evolved by the virus to escape anti-Env neutralizing Abs. This may explain the low effectiveness of Env-based vaccines, which are also unlikely to elicit Abs against new Env epitopes exposed by the Tat/Env interaction. As Tat also binds Envs from different clades, new vaccine strategies should exploit the Tat/Env interaction for both preventative and therapeutic interventions.

Interferon alpha (IFNalpha) induces both apoptosis and a counteracting epidermal growth factor Erk-dependent survival response in cancer cells. In this report, IFNalpha increased eukaryotic elongation factor 1A (eEF-1A) protein expression by inhibition of eEF-1A degradation via a proteasome-dependent pathway. The reduction of the expression level of eEF-1A by RNA interference enhanced the apoptosis induced by IFNalpha on the same cells. Moreover, IFNalpha induced the phosphorylation of both serine and threonine in eEF-1A. These effects were paralleled by an increased co-immunoprecipitation and colocalization of eEF-1A with C-Raf. The suppression of C-Raf kinase activity with the inhibitor BAY 43-9006 completely antagonized the increase of both eEF-1A phosphorylation and expression and of C-Raf/eEF-1A colocalization induced by IFNalpha and enhanced apoptosis and eEF-1A ubiquitination. Cell transfection with the mutated K48R ubiquitin increased EF-1A expression and desensitized tumor cells to the modulating effects of IFNalpha. The dynamic simulation of 3Dstructure of eEF-1A identified putative serine and threonine phosphorylation sites. In conclusion, the interaction between eEF-1A and C-Raf increases eEF-1A stability and induces a survival activity.

Il waterfront è un luogo particolare sia per la città che per il progetto di architettura. Esistono due modi per intenderlo: uno perpendicolare ad esso ed uno orizzontale. Il primo è quello più tradizionale e scontato, il secondo invece meno diffuso ma di forte interesse.

Il tema del rapporto tra architettura e natura e’ stato occasione secolare di esplorazione e ricerca. Nelle sue declinazioni piu recenti questo rapporto ha osservato una sempre maggiore integrazione tra concetti che possono apparire antinomici. Il bisogno di Natura in Architettura ottiene una consacrazione con il pensiero di Le Corbusier: il tetto-giardino prima, esplicitazione della integrazione della natura in architettura, che mette sull’edificio moderno la dimensione naturale normalmente destinata al livello del giardino. Successivamente gli studi sulla citta’ moderna e la sua integrazione con i sistemi naturali e il verde fanno di Le Corbusier un paesaggista innovatore; con la Ville Contemporaine per tre milioni di abitanti (1922), Il Plano Voisin per Parigi (1925), e la Ville Radieuse per un milione e mezzo di abitanti (1929), Le Corbusier trasforma in chiave contemporanea e dirompente le teorizzazioni di Howard sulla Città-Giardino, delineando in forma ipermoderna il nuovo rapporto tra natura e sistemi urbani, la cui qualità genera una totale integrazione tra artificio e natura, disciplinando altresí una nuova gerarchia negli spazi pubblici.Il saggio esplora esperienze di trasformazioni del paesaggio urbano.

Summary Background We ran a compassionate use nationwide programme (ABACUS) to provide access to ombitasvir, paritaprevir, and ritonavir, with dasabuvir, plus ribavirin for hepatitis C virus (HCV) genotype 1 infection and ombitasvir, paritaprevir, and ritonavir, plus ribavirin for HCV genotype 4 infection in patients with cirrhosis at high risk of decompensation while approval of these regimens was pending in Italy. Methods In this prospective observational study, we collected data from a compassionate use nationwide programme from March 17, 2014, to May 28, 2015. Patients with HCV genotype 1 infection and cirrhosis at high risk of decompensation were given coformulated ombitasvir (25 mg), paritaprevir (150 mg), and ritonavir (100 mg) once daily and dasabuvir (250 mg) twice daily for 12 weeks (patients with HCV genotype 1b infection) or 24 weeks (patients with HCV genotype 1a infection). Patients with HCV genotype 4 infection were given coformulated ombitasvir (25 mg), paritaprevir (150 mg), and ritonavir (100 mg) once per day for 24 weeks. All patients were given weight-based ribavirin. The primary efficacy endpoint was sustained virological response at week 12 after the end of treatment (SVR12), analysed by intention-to-treat. Univariate and multivariate logistic regression analyses were used to identify baseline characteristics associated with SVR12. Adverse events were recorded throughout the study. Findings 728 (96%) of 762 patients with cirrhosis who were given ombitasvir, paritaprevir, and ritonavir, with or without dasabuvir, plus ribavirin therapy for 12 or 24 weeks achieved SVR12. Logistic regression analyses identified that bilirubin concentrations of less than 2 mg/dL were associated with SVR12 (odds ratio [OR] 4·76 [95% CI 1·83–12·3]; p=0·001). 166 (23%) of 734 patients included in safety analyses had an adverse event. 25 (3%) patients discontinued treatment because of adverse events. Asthenia was the most commonly reported adverse event, occurring in 36 (5%) patients. Interpretation Our findings suggest that the safety and effectiveness of ombitasvir, paritaprevir, and ritonavir, with or without dasabuvir, plus ribavirin in patients with HCV genotype 1 or 4 infection and cirrhosis at high risk of decompensation in a real-life setting are similar to those reported in clinical trials. The concordance with clinical trials provides reassurance that the reported efficacy of this treatment in clinical trials will translate to its use in routine clinical practice. Funding Dipartimento Biomedico di Medicina Interna e Specialistica dell'Universita di Palermo.

Background The phase II multicenter, randomized, open label, therapeutic trial (ISS T-002, Clinicaltrials.gov NCT00751595) was aimed at evaluating the immunogenicity and the safety of the biologically active HIV-1 Tat protein administered at 7.5 or 30 μg, given 3 or 5 times monthly, and at exploring immunological and virological disease biomarkers. The study duration was 48 weeks, however, vaccinees were followed until the last enrolled subject reached the 48 weeks. Reported are final data up to 144 weeks of follow-up. The ISS T-002 trial was conducted in 11 clinical centers in Italy on 168 HIV positive subjects under Highly Active Antiretroviral Therapy (HAART), anti-Tat Antibody (Ab) negative at baseline, with plasma viremia

Background: Tat is a key HIV-1 virulence factor, which plays pivotal roles in virus gene expression, replication, transmission and disease progression. After release, extracellular Tat accumulates in tissues and exerts effects on both the virus and the immune system, promoting immune activation and virus spreading while disabling the host immune defense. In particular, Tat binds Env spikes on virus particles forming a virus entry complex, which favors infection of dendritic cells and efficient transmission to T cells via RGD-binding integrins. Tat also shields the CCR5-binding sites of Env rendering ineffective virus neutralization by anti-Env antibodies (Abs). This is reversed by the anti-Tat Abs present in natural infection or induced by vaccination.Findings: Here we present the results of a cohort study, showing that the presence of anti-Tat Abs in asymptomatic and treatment-naïve HIV-infected subjects is associated with containment of CD4+ T-cell loss and viral load and with a delay of disease progression. In fact, no subjects with high anti-Tat Ab titers initiated antiretroviral therapy during the three years of follow-up. In contrast, no significant effects were seen for anti-Env and anti-Gag Abs. The increase of anti-Env Ab titers was associated with a reduced risk of starting therapy only in the presence of anti-Tat Abs, suggesting an effect of combined anti-Tat and anti-Env Abs on the Tat/Env virus entry complex and on virus neutralization.Conclusions: Anti-Tat immunity may help delay HIV disease progression, thus, targeting Tat may offer a novel therapeutic intervention to postpone antiretroviral treatment or to increase its efficacy. © 2014 Bellino et al.; licensee BioMed Central Ltd

Objective:The identification of still unrevealed mechanisms affecting the anti-HIV CD8(+) T-cell response in HIV-1 infection.Design:Starting from the observation that anti-Tat immunization is associated with improved CD8(+) T-cell immunity, we developed both in-vitro and ex-vivo assays to characterize the effects of extra-cellular Tat on the adaptive CD8(+) T-cell response.Methods:The effects of Tat on CD8(+) T-cell activation were assayed using CD8(+) T-cell clones specific for either cellular (MART-1) or viral (HIV-1 Nef) antigens, and HIV-1 Gag-specific CD8(+) T cells from HIV-1 patients.Results:The interaction between CD8(+) T lymphocytes and immobilized Tat, but not its soluble form, inhibits peptide-specific CD8(+) T-lymphocyte activation. The inhibition does not depend on Tat trans-activation activity, but on the interaction of the Tat RGD domain with 51 and v3 integrins. Impaired CD8(+) T-cell activation was also observed in cocultures of CD8(+) T cells with HIV-1-infected cells. Anti-Tat Abs abrogate the inhibitory effect, consistently with the evidence that extracellular Tat accumulates on the cell membrane of virus-producing cells. The Tat-induced inhibition of cell activation associates with increased apoptosis of CD8(+) T cells. Finally, the inhibition of cell activation also takes place in Gag-specific CD8(+) T lymphocytes from HIV-1-infected patients.Conclusion:Our results support the idea that CD8(+) T-cell apoptosis induced by surface-bound extracellular Tat can contribute to the dysregulation of the CD8(+) T-cell adaptive response against HIV as well as other pathogens present in AIDS patients.

The stepwise chromatographic behaviour on DEAE-Sepharose of rat Fe65, a neuronal protein, was tested, using as eluants KCl, CaCl2, and MgCl2. Assays by western blot showed that Fe65 was eluted by CaCl2, at a ionic strength 20% lower than that of MgCl2 or KCl. Interestingly, in the case of a truncated Fe65, lacking a glutamic acid rich region at the N-terminus, the ionic strengths of the various eluants were almost identical. These results suggested a possible inhibitory role of calcium ions in the binding of the protein to DEAE and a specific affinity of these ions for long acidic stretches.

An increasing number of evidences suggest the involvement of the eukaryotic elongation factor 1A, a core component of the protein synthesis machinery, at the onset of cell transformation. In fact, eEF1A is shown to be up-regulated in cell death; moreover, it seems to be involved in the regulation of ubiquitin-mediated protein degradation. In addition, eEF1A undergoes several post-translational modifications, mainly phosphorylation and methylation, that generally influence the activity of the protein. This article summarizes the present knowledges on the several extra-translational roles of eEF1A also in order to understand as the protein synthesis regulatory mechanisms could offer tools for cancer intervention.

1999/2000 XIII Ciclo 1970 Versione digitalizzata della tesi di dottorato cartacea.

1998/1999 Negli ultimi anni passati le applicazioni multimediali hanno visto uno sviluppo notevole, trovando applicazione in un gran numero di campi. Applicazioni come video conferenze, diagnostica medica, telefonia mobile e applicazioni militari necessitano il trattamento di una gran mole di dati ad alta velocità. Pertanto, l'elaborazione di immagini e di dati vocali è molto importante ed è stata oggetto di numerosi sforzi, nel tentativo di trovare algoritmi sempre più veloci ed efficaci. Tra gli algoritmi proposti, noi crediamo che gli operatori razionali svolgano un ruolo molto importante, grazie alla loro versatilità ed efficacia nell'elaborazione di dati. Negli ultimi anni sono stati proposti diversi algoritmi, dimostrando che questi operatori possono essere molto vantaggiosi in diverse applicazioni, producendo buoni risultati. Lo scopo di questo lavoro è di realizzare alcuni di questi algoritmi e, quindi, dimostrare che i filtri razionali, in particolare, possono essere realizzati senza ricorrere a sistemi di grandi dimensioni e possono raggiungere frequenze operative molto alte. Una volta che il blocco fondamentale di un sistema basato su operatori razionali sia stato realizzato, esso pu6 essere riusato con successo in molte altre applicazioni. Dal punto di vista del progettista, è importante avere uno schema generale di studio, che lo renda capace di studiare le varie configurazioni del sistema da realizzare e di analizzare i compromessi tra le variabili di progetto. In particolare, per soddisfare l'esigenza di metodi versatili per la stima della potenza, abbiamo sviluppato una tecnica di macro modellizazione che permette al progettista di stimare velocemente ed accuratamente la potenza dissipata da un circuito. La tesi è organizzata come segue: Nel Capitolo 1 alcuni sono presentati alcuni algoritmi studiati per la realizzazione. Ne viene data solo una veloce descrizione, lasciando comunque al lettore interessato dei riferimenti bibliografici. Nel Capitolo 2 vengono discusse le architetture fondamentali usate per la realizzazione. Principalmente sono state usate architetture a pipeline, ma viene data anche una descrizione degli approcci oggigiorno disponibili per l'ottimizzazione delle temporizzazioni. Nel Capitolo 3 sono presentate le realizzazioni di due sistemi studiati per questa tesi. Gli approcci seguiti si basano su ASIC e FPGA. Richiedono tecniche e soluzioni diverse per il progetto del sistema, per cui é interessante vedere cosa pu6 essere fatto nei due casi. Infine, nel Capitolo 4, descriviamo la nostra tecnica di macro modellizazione per la stima di potenza, dando una breve visione delle tecniche finora proposte e facendo vedere quali sono i vantaggi che il nostro metodo comporta per il progetto. In the past few years, multimedia application have been growing very fast, being applied to a large variety of fields. Applications like video conference, medical diagnostic, mobile phones, military applications require to handle large amount of data at high rate. Images as well as voice data processing are therefore very important and they have been subjected to a lot of efforts in order to find always faster and effective algorithms. Among image processing algorithms, we believe that rational operators assume an important role, due to their versatility and effectiveness in data processing. In the last years, several algorithms have been proposed, demonstrating that these operators can be very suitable in different applications with very good results. The aim of this work is to implement some of these algorithm and, therefore, demonstrate that rational filters, in particular, can be implemented without requiring large sized systems and they can operate at very high frequencies. Once the basic building block of a rational based system has been implemented, it can be successfully reused in many other applications. From the designer point of view, it is important to have a general framework, which makes it able to study various configurations of the system to be implemented and analyse the trade-off among the design variables. In particular, to meet the need far versatile tools far power estimation, we developed a new macro modelling technique, which allows the designer to estimate the power dissipated by a circuit quickly and accurately. The thesis is organized as follows: In chapter 1 we present some of the algorithms which have been studied for implementation. Only a brief overview is given, leaving to the interested reader some references in literature. In chapter 2 we discuss the basic architectures used for the implementations. Pipelined structures have been mainly used for this thesis, but an overview of the nowaday available approaches for timing optimization is presented. In chapter 3 we present two of the implementation designed for this thesis. The approaches followed are ASIC driven and FPGA drive. They require different techniques and different solution for the design of the system, therefore it is interesting to see what can be done in both the cases. Finally, in chapter 4, we describe our macro modelling techniques for power estimation, giving a brief overview of the up to now proposed techniques and showing the advantages our method brings to the design. XII Ciclo 1969 Versione digitalizzata della tesi di dottorato cartacea.

1997/1998 Questa tesi, intitolata Visione Tridimensionale per la stima di Struttura e Moto, tratta di tecniche di Visione Artificiale per la stima delle proprietà geometriche del mondo tridimensionale a partire da immagini numeriche. Queste proprietà sono essenziali per il riconoscimento e la classificazione di oggetti, la navigazione di veicoli mobili autonomi, il reverse engineering e la sintesi di ambienti virtuali. In particolare, saranno descritti i moduli coinvolti nel calcolo della struttura della scena a partire dalle immagini, e verranno presentati contributi originali nei seguenti campi. Rettificazione di immagini steroscopiche. Viene presentato un nuovo algoritmo per la rettificazione, il quale trasforma una coppia di immagini stereoscopiche in maniera che punti corrispondenti giacciano su linee orizzontali con lo stesso indice. Prove sperimentali dimostrano il corretto comportamento del metodo, come pure la trascurabile perdita di accuratezza nella ricostruzione tridimensionale quando questa sia ottenuta direttamente dalle immagini rettificate. Calcolo delle corrispondenze in immagini stereoscopiche. Viene analizzato il problema della stereovisione e viene presentato un un nuovo ed efficiente algoritmo per l'identificazione di coppie di punti corrispondenti, capace di calcolare in modo robusto la disparità stereoscopica anche in presenza di occlusioni. L'algoritmo, chiamato SMW, usa uno schema multi-finestra adattativo assieme al controllo di coerenza destra-sinistra per calcolare la disparità e l'incertezza associata. Gli esperimenti condotti con immagini sintetiche e reali mostrano che SMW sortisce un miglioramento in accuratezza ed efficienza rispetto a metodi simili Inseguimento di punti salienti. L'inseguitore di punti salienti di Shi-Tomasi- Kanade viene migliorato introducendo uno schema automatico per lo scarto di punti spuri basato sulla diagnostica robusta dei campioni periferici ( outliers ). Gli esperimenti con immagini sintetiche e reali confermano il miglioramento rispetto al metodo originale, sia qualitativamente che quantitativamente. Ricostruzione non calibrata. Viene presentata una rassegna ragionata dei metodi per la ricostruzione di un modello tridimensionale della scena, a partire da una telecamera che si muove liberamente e di cui non sono noti i parametri interni. Il contributo consiste nel fornire una visione critica e unificata delle più recenti tecniche. Una tale rassegna non esiste ancora in letterarura. Moto tridimensionale. Viene proposto un algoritmo robusto per registrate e calcolare le corrispondenze in due insiemi di punti tridimensionali nei quali vi sia un numero significativo di elementi mancanti. Il metodo, chiamato RICP, sfrutta la stima robusta con la Minima Mediana dei Quadrati per eliminare l'effetto dei campioni periferici. Il confronto sperimentale con una tecnica simile, ICP, mostra la superiore robustezza e affidabilità di RICP. This thesis addresses computer vision techniques estimating geometrie properties of the 3-D world /rom digital images. Such properties are essential for object recognition and classification, mobile robots navigation, reverse engineering and synthesis of virtual environments. In particular, this thesis describes the modules involved in the computation of the structure of a scene given some images, and offers original contributions in the following fields. Stereo pairs rectification. A novel rectification algorithm is presented, which transform a stereo pair in such a way that corresponding points in the two images lie on horizontal lines with the same index. Experimental tests prove the correct behavior of the method, as well as the negligible decrease oLthe accuracy of 3-D reconstruction if performed from the rectified images directly. Stereo matching. The problem of computational stereopsis is analyzed, and a new, efficient stereo matching algorithm addressing robust disparity estimation in the presence of occlusions is presented. The algorithm, called SMW, is an adaptive, multi-window scheme using left-right consistency to compute disparity and its associated uncertainty. Experiments with both synthetic and real stereo pairs show how SMW improves on closely related techniques for both accuracy and efficiency. Features tracking. The Shi-Tomasi-Kanade feature tracker is improved by introducing an automatic scheme for rejecting spurious features, based on robust outlier diagnostics. Experiments with real and synthetic images confirm the improvement over the original tracker, both qualitatively and quantitatively. 111 Uncalibrated vision. A review on techniques for computing a three-dimensional model of a scene from a single moving camera, with unconstrained motion and unknown parameters is presented. The contribution is to give a critical, unified view of some of the most promising techniques. Such review does not yet exist in the literature. 3-D motion. A robust algorithm for registering and finding correspondences in two sets of 3-D points with significant percentages of missing data is proposed. The method, called RICP, exploits LMedS robust estimation to withstand the effect of outliers. Experimental comparison with a closely related technique, ICP, shows RICP's superior robustness and reliability. XI Ciclo 1968 Versione digitalizzata della tesi di dottorato cartacea.

1996/1997 X Ciclo 1967 Versione digitalizzata della tesi di dottorato cartacea.