Selezione e processazione di cellule Natural Killer da donatore aploidentico "KIR-ligand" incompatibile per l'immunoterapia adottiva di pazienti con Leucemia Acuta Mieloblastica ad alto rischio

The effector function of natural killer (NK) cells is regulated by activating and inhibitory receptors, termed killer immunoglobulin-like receptors (KIRs). In haploidentical T-cell depleted transplantation the donor/recipient KIR mismatch significantly impacts on NK-mediated tumor cell killing, particularly in acute myeloid leukaemia (AML). Thirty-four high risk AML patients entered a phase I-II study of adoptive NK-cell based immunotherapy and were screened for the availability of one haploidentical KIR ligand mismatched donor. Thirteen of them resulted as having one suitable donor. NK cells were enriched from steady-state leukaphereses by using a double-step immunomagnetic separation system, consisting in depletion of CD3+ T cells followed by positive selection of CD56+ NK cells. CD56+ cells were enriched from 7,70% (1,26-11,70) to 93,50% (66,41-99,20) (median recovery 53,05% (30,97-72,85), median T-depletion 3,03 log (2,15-4,52) viability >92%) and their citotoxic activity was inalterate. All patients (4 progressions, 1 partial remission and 8 complete remissions) received NK cell infusion which was preceeded by immunosuppressive chemotherapy (fludarabine and cyclophosphamide) and followed by interleukin 2 injections. The median number of reinfused NK cells was 2,74x10(e)6/kg(1,11-5,00) and contamining CD3+ T cells were always less than 1x10(e)5/kg. The procedure was well-tolerated and no significant toxicity, including GvHD, related to NK cell infusion was observed. The donor NK cells were demonstrated in 5/10 patients. Among the 8 patients in complete remission 5 patients are stable after 18, 15, 4, 2 months of follow-up. Three other patients relapsed after 2 and 7 months. The patient in partial remission obtained a complete remission, which lasted for 6 months. The 4 patients with active/progressive disease showed the persistence of disease. This clinical observation may be correlated with in vitro studies, indicating that AML cells are capable to induce NK cell apoptosis in a dose-depend manner. In summery, a two-step enrichment of CD56+ NK cells allows the collection of a suitable number of target cells to be used as adoptive immunotherapy in AML patients. Infusion of NK cells is feasible and safe and adoptively transferred NK cells can be detected after infusion.

Costrutti ed Applicazioni di Programmazione Generica in Linguaggi Object-Oriented

Generic programming is likely to become a new challenge for a critical mass of developers. Therefore, it is crucial to refine the support for generic programming in mainstream Object-Oriented languages — both at the design and at the implementation level — as well as to suggest novel ways to exploit the additional degree of expressiveness made available by genericity. This study is meant to provide a contribution towards bringing Java genericity to a more mature stage with respect to mainstream programming practice, by increasing the effectiveness of its implementation, and by revealing its full expressive power in real world scenario. With respect to the current research setting, the main contribution of the thesis is twofold. First, we propose a revised implementation for Java generics that greatly increases the expressiveness of the Java platform by adding reification support for generic types. Secondly, we show how Java genericity can be leveraged in a real world case-study in the context of the multi-paradigm language integration. Several approaches have been proposed in order to overcome the lack of reification of generic types in the Java programming language. Existing approaches tackle the problem of reification of generic types by defining new translation techniques which would allow for a runtime representation of generics and wildcards. Unfortunately most approaches suffer from several problems: heterogeneous translations are known to be problematic when considering reification of generic methods and wildcards. On the other hand, more sophisticated techniques requiring changes in the Java runtime, supports reified generics through a true language extension (where clauses) so that backward compatibility is compromised. In this thesis we develop a sophisticated type-passing technique for addressing the problem of reification of generic types in the Java programming language; this approach — first pioneered by the so called EGO translator — is here turned into a full-blown solution which reifies generic types inside the Java Virtual Machine (JVM) itself, thus overcoming both performance penalties and compatibility issues of the original EGO translator. Java-Prolog integration Integrating Object-Oriented and declarative programming has been the subject of several researches and corresponding technologies. Such proposals come in two flavours, either attempting at joining the two paradigms, or simply providing an interface library for accessing Prolog declarative features from a mainstream Object-Oriented languages such as Java. Both solutions have however drawbacks: in the case of hybrid languages featuring both Object-Oriented and logic traits, such resulting language is typically too complex, thus making mainstream application development an harder task; in the case of library-based integration approaches there is no true language integration, and some “boilerplate code” has to be implemented to fix the paradigm mismatch. In this thesis we develop a framework called PatJ which promotes seamless exploitation of Prolog programming in Java. A sophisticated usage of generics/wildcards allows to define a precise mapping between Object-Oriented and declarative features. PatJ defines a hierarchy of classes where the bidirectional semantics of Prolog terms is modelled directly at the level of the Java generic type-system.

Strategies for detecting DNA hybridisation using surface plasmon fluorescence spectroscopy

Die Anregung und Emission von Fluorophoren nahe planaren Metalloberflächen und schiefen Gittern wurde mittels Oberflächenplasmonen Fluoreszenz Spektroskopie (SPFS) untersucht. Die Fluorophore konnten durch das evaneszente Plasmonenfeld angeregt und die einzelnen Abregungskanäle identifiziert werden.Die Sensorarchitektur für den Nachweis der Hybridisierung bestand aus auf einer Streptavidin-Matrix immobilisierten unmarkierten Sondensträngen. Cy5 markierte Zielsequenzen wurden aus der Lösung hybridisiert und die Adsorptionskinetiken konnten oberflächensensitiv detektiert werden.Ein neues Detektionsschema für unmarkierte Zielstränge wurde mittels fluoreszenzmarkirten Sondensträngen realisiert. Die Hybridisierung führte zu der Bildung von steifen helikalen Bereichen in der Probe und separierte den Farbstoff von der Metalloberfläche. Reduzierte Fluorezenzlöschung zeigte daher das Hybridisierungsereignis an.Die Verwendung eines potentiellen Förster-Paares zur Detektion von DNA Hybridisierung wurde untersucht. Donor und Akzeptor wurden an Ziel- und Sondenstrang immobilisiert und das Hybridisierungsereignis konnte anhand der Auslöschung der Donor-Fluorezenz nachgewiesen werden.Schließlich wurde der Einsatz von einzelstrangbindenden Proteinen (SSB) zur Steigerung der Sensitivität bezüglich Basenfehlpaarungen betrachtet. Verdrängungsreaktionen zwischen Proteinen und markierten Zielsträngen wurden anhand von SPS und Fluorezenzkinetiken studiert.

X-ray and lensing mass estimates in galaxy clusters

In this Thesis we have presented our work on the analysis of galaxy clusters through their X-ray emission and the gravitational lensing effect that they induce. Our research work was mainly finalised to verify and possibly explain the observed mismatch between the galaxy cluster mass distributions estimated through two of the most promising techniques, i.e. the X-ray and the gravitational lensing analyses. Moreover, it is an established evidence that combined, multi-wavelength analyses are extremely effective in addressing and explaining the open issues in astronomy: however, in order to follow this approach, it is crucial to test the reliability and the limitations of the individual analysis techniques. In this Thesis we also assessed the impact of some factors that could affect both the X-ray and the strong lensing analyses.

Thin functional conducting polymer films

In the present study, thin functional conducting polyaniline (PANI) films, either doped or undoped, patterned or unpatterned, were prepared by different approaches. The properties of the obtained PANI films were investigated in detail by a combination of electrochemistry with several other techniques, such as SPR, QCM, SPFS, diffraction, etc. The sensing applications (especially biosensing applications) of the prepared PANI films were explored. Firstly, the pure PANI films were prepared by the electropolymerisation method and their doping/dedoping properties in acidic conditions were investigated in detail by a combination of electrochemistry with SPR and QCM. Dielectric constants of PANI at different oxidation states were obtained quantitatively. The results obtained here laid a good foundation for the following investigations of PANI films in neutral pH conditions. Next, PANI multilayer films doped by a variety of materials were prepared by the layer-by-layer method in order to explore their biosensing applications, because of the loss of redox activity of pure PANI in neutral pH conditions. The dopants used include not only the traditionally used linear polyelectrolytes, but also, for the first tim, some other novel materials, like modified gold nanoparticles or modified carbon nanotubes. Our results showed that all the used dopants could form stable multilayer films with PANI. All the obtained PANI multilayer films showed good redox activity in a neutral pH environment, which makes them feasible for bioassays. We found that all the prepared PANI multilayer films can electrocatalyze the oxidation of NADH in neutral conditions at a low potential, although their catalytic efficiencies are different. Among them, PANI/carbon nanotube system showed the highest catalytic efficiency toward the oxidation of NADH, which makes it a good candidate as a NADH sensor. Besides, because some of the prepared PANI multilayer systems were end-terminated with –COOH groups (like PANI/Au nanoparticles system), which can be utilized to easily link biomolecules for biosensing applications. Here, we demonstrated, for the first time, to use the prepared PANI multilayer films for the DNA hybridisation detection. The detection event was monitored either by direct electrochemical method, or by enzyme-amplified electrochemical method, or by surface plasmon enhanced fluorescence spectroscopic method. All the methods can effectively differentiate non-complementary DNA from the complementary ones, even at the single-base mismatch level. It should also be noted that, our success in fabricating PANI multilayer films with modified Au nanoparticles or carbon nanotubes also offered another novel method for incorporating such novel materials into (conducting) polymers. Because of the unique electrochemical and optical properties of each component of the obtained PANI multilayer films, they should also find potential applications in many other fields such as microelectronics, or for electrochromic and photovoltaic devices. Finally, patterned PANI films were fabricated by the combination of several patterning techniques, such as the combination of electrocopolymerization with micromolding in capillaries (EP-MIMIC), the combination of microcontact printing with the layer-by-layer technique (µCP-LBL), and the polystyrene (PS) template induced electropolymerisation method. Using the obtained stripe-shaped PANI/PSS film, a redox-switchable polymer grating based on the surface-plasmon-enhanced mode was constructed and its application in the field of biosensing was explored. It was found that the diffraction efficiency (DE) of the grating was very sensitive to the applied potential (i.e. redox state of the film) as well as the pH environment of the dielectric medium. Moreover, the DE could also be effectively tuned by an electrocatalytic event, such as the electrocatalytic oxidation of NADH by the grating film. By using PS colloidal crystal assemblies as templates, well-ordered 3D interconnected macroporous PANI arrays (PANI inverse opals) were fabricated via electropolymerisation method. The quality of the obtained inverse opals was much higher than those reported by chemical synthesis method. By electrochemical method, the structures of the prepared inverse opals can be easily controlled. To explore the possible biosensing applications of PANI inverse opals, efforts were also done toward the fabrication of PANI composite inverse opals. By selecting proper dopants, high quality inverse opals of PANI composites were fabricated for the first time. And the obtained opaline films remained redox-active in neutral pH conditions, pointing to their possible applications for electrobioassays.

Kinetic and affinity analysis of hybridization reactions between PNA probes and DNA targets using surface plasmon field-enhanced fluorescence spectroscopy (SPFS)

Sequenz spezifische biomolekulare Analyseverfahren erweisen sich gerade im Hinblick auf das Humane Genom Projekt als äußerst nützlich in der Detektion von einzelnen Nukleotid Polymorphismen (SNPs) und zur Identifizierung von Genen. Auf Grund der hohen Anzahl von Basenpaaren, die zu analysieren sind, werden sensitive und effiziente Rastermethoden benötigt, welche dazu fähig sind, DNA-Proben in einer geeigneten Art und Weise zu bearbeiten. Die meisten Detektionsarten berücksichtigen die Interaktion einer verankerten Probe und des korrespondierenden Targets mit den Oberflächen. Die Analyse des kinetischen Verhaltens der Oligonukleotide auf der Sensoroberfläche ist infolgedessen von höchster Wichtigkeit für die Verbesserung bereits bekannter Detektions - Schemata. In letzter Zeit wurde die Oberflächen Plasmonen feld-verstärkte Fluoreszenz Spektroskopie (SPFS) entwickelt. Sie stellt eine kinetische Analyse - und Detektions - Methode dar, die mit doppelter Aufzeichnung, d.h. der Änderung der Reflektivität und des Fluoreszenzsignals, für das Interphasen Phänomen operiert. Durch die Verwendung von SPFS können Kinetikmessungen für die Hybridisierung zwischen Peptid Nukleinsäure (PNA), welche eine synthetisierte Nukleinsäure DNA imitiert und eine stabilere Doppelhelix formt, und DNA auf der Sensoroberfläche ausgeführt werden. Mittels einzel-, umfassend-, und titrations- Experimenten sowohl mit einer komplementär zusammenpassenden Sequenz als auch einer mismatch Sequenz können basierend auf dem Langmuir Modell die Geschwindigkeitskonstanten für die Bindungsreaktion des oligomer DNA Targets bzw. des PCR Targets zur PNA ermittelt werden. Darüber hinaus wurden die Einflüsse der Ionenstärke und der Temperatur für die PNA/DNA Hybridisierung in einer kinetischen Analyse aufgezeigt.

Special Heusler compounds for spintronic applications

This work emphasizes the potential of Heusler compounds in a wide range of spintronic applications. Using electronic structure calculations it is possible to design compounds for specific applications. Examples for GMR and TMR applications, for spin injection into semiconductors, and for spin torque transfer applications will be shown. After a detailed introduction about spintronics and related materials chapter 5 reports about the investigation of new half-metallic compounds where the Fermi energy is tuned in the middle of the gap to result in more stable compounds for GMR and TMR applications. The bulk properties of the quaternary Heusler alloy Co2Mn(1-x)Fe(x)Si with the Fe concentration ranging from x=0 to 1 will be reported and the results suggest that the best candidate for applications may be found at an iron concentration of about 50%. Due to the effect that in the Co2Mn(1-x)Fe(x)Si series the transition metal carrying the localized moment is exchanged and this might lead to unexpected effects on the magnetic properties if the samples are not completely homogeneous chapter 6 reports about the optimization of the Heusler compounds for GMR and TMR applications. The structural and magnetic properties of the quaternary Heusler alloy Co2FeAl(1-x)Si(x) with varying Si concentration will be reported. From the combination of experimental (better order for high Si content) and theoretical findings (robust gap at x = 0.5) it is concluded that a compound with an intermediate Si concentration close to x=0.5-0.7 would be best suited for spintronic applications, especially for GMR and TMR applications. In chapter 7 the detailed investigation of compounds for spin injection into semiconductors will be reported. It will be shown that the diluted magnetic semiconductors based on CoTiSb with a very low lattice mismatch among each other are interesting materials for spintronics applications like Spin-LEDs or other spin injection devices. Chapter 8 refers about the investigation of the theoretically predicted half-metallic completely compensated-ferrimagnet Mn$_3$Ga as a suitable material for spin torque transfer applications. The Curie temperature is above 730~K and the electronic structure calculations indicate a nearly half-metallic ferrimagnetic order with 88% spin polarization at the Fermi energy.}

La molecola IRTA1 (immunoglobulin superfamily receptor trans location associated 1) risulta selettivamente espressa nei linfomi non Hodgkin B della zona marginale

La diagnosi di linfoma non Hodgkin B della zona marginale si basa su criteri morfologici e sulla sostanziale negatività per marcatori immunoistochimici espressi in altri sottotipi di linfoma B. L’ obiettivo di questo lavoro è stato, quindi, quello di ricercare una molecola specifica associata ai linfomi della zona marginale. Materiali e Metodi. Sono stati esaminati 2.104 linfomi periferici di entità nosologia eterogenea mediante un anticorpo monoclonale, diretto contro la molecola IRTA1, che riconosce la zona marginale nei tessuti linfoidi umani. Risultati. Si è riscontrata espressione di IRTA1 nel 93% dei linfomi della zona marginale ad insorgenza extranodale e nel 74% di quelli primitivi linfonodali suggerendo la possibilità che questi linfomi possano originare dalle cellule perifollicolari o monocitoidi IRTA1+ riscontrabili nei linfonodi reattivi. La valutazione immunoistochimica mediante doppia colorazione (IRTA1/bcl6), ha inoltre dimostrato come vi sia una modulazione fenotipica nelle cellule marginali neoplastiche nel momento in cui esse colonizzano i follicoli linfoidi e durante la loro circolazione nei centri germinativi. Le cellule marginali neoplastiche che differenziano in senso plasmacellulare perdono l’ espressione di IRTA1 Discussione. In conclusione, tali evidenze hanno permesso di ampliare la conoscenza sulla biologia dei linfomi marginali e sottolineano come IRTA1 sia il primo marcatore diagnostico positivo per queste neoplasie.

I limiti al diritto di proprietà derivanti dagli strumenti di pianificazione urbanistica. I vincoli urbanistici.

Nella prima parte viene ricostruito il concetto di vincolo espropriativo alla luce dell’elaborazione della giurisprudenza della Corte costituzionale e della Corte EDU, giungendo alla conclusione che rientrano in tale concetto le limitazioni al diritto di proprietà che: - derivano da scelte discrezionali dell’Amministrazione non correlate alle caratteristiche oggettive del bene; - superano la normale tollerabilità nel senso che impediscono al proprietario la prosecuzione dell’uso in essere o incidono sul valore di mercato del bene in modo sproporzionato rispetto alle oggettive caratteristiche del bene e all’interesse pubblico perseguito. Ragione di fondo della teoria dei vincoli è censurare l’eccessiva discrezionalità del potere urbanistico, imponendo una maggiore obiettività e controllabilità delle scelte urbanistiche. Dalla teoria dei vincoli consegue altresì che nell’esercizio del potere urbanistico l’Amministrazione, pur potendo differenziare il territorio, deve perseguire l’obiettivo del riequilibrio economico degli interessi incisi dalle sue determinazioni. L’obbligo della corresponsione dell’indennizzo costituisce la prima forma di perequazione urbanistica. Nel terzo e nel quarto capitolo viene analizzata la giurisprudenza civile e amministrativa in tema di vincoli urbanistici, rilevandone la non corrispondenza rispetto all’elaborazione della Corte costituzionale e l’incongruità dei risultati applicativi. Si evidenzia in particolare la necessità del superamento del criterio basato sulla distinzione zonizzazioni-localizzazioni e di considerare conformative unicamente quelle destinazioni realizzabili ad iniziativa privata che in concreto consentano al proprietario di conseguire un’utilità economica proporzionata al valore di mercato del bene. Nel quinto capitolo viene analizzato il rapporto tra teoria dei vincoli e perequazione urbanistica, individuandosi il discrimine tra i due diversi istituti non solo nel consenso, ma anche nella proporzionalità delle reciproche prestazioni negoziali. Attraverso la perequazione non può essere attribuito al proprietario un’utilità inferiore a quella che gli deriverebbe dall’indennità di esproprio.

Untersuchungen zum Mechanismus der Zytotoxizität von alkylierenden Agenzien in malignen Melanomzellen

Maligne Melanome sind gegenüber Chemotherapeutika relativ resistent. Das methylierende Alkylanz Temozolomid sowie das chlorethylierende und DNA-Interstrand Crosslink (ICL) bildende Alkylanz Fotemustin kommen bei der Behandlung des malignen Melanoms als Mittel erster Wahl zum Einsatz. In der vorliegenden Arbeit konnte das erste Mal nachgewiesen werden, dass die zytotoxische Wirkung von Temozolomid und Fotemustin in Melanomzellen durch Apoptose vermittelt wird. Unter Verwendung klinisch relevanter Dosen der beiden Alkylantien konnte die Induktion von Apoptose durch vier unabhängige Methoden (Bestimmung der SubG1-Fraktion und der Apoptose- / Nekrose-Frequenz, Aktivierung der Effektorcaspasen-3 und -7 sowie Spaltung von PARP-1) nachgewiesen werden. Die Alkylierungen an der O6-Position des Guanins, welche durch beide Agenzien induziert werden, sind auch in Melanomzellen die wichtigsten Zytotoxizität-bewirkenden Läsionen in der DNA, und die O6-Methylguanin-DNA-Methyltransferase (MGMT) ist folglich ein herausragender Resistenzmarker. Eine der verwendeten Zelllinien (D05) exprimierte p53-Wildtypprotein. Diese Zelllinie war resistenter als alle anderen Zelllinien gegenüber Temozolomid und Fotemustin. Dies weist darauf hin, dass p53 nicht die Apoptoseinduktion in Melanomzellen verstärkt. Die Prozessierung des O6MeG erfolgt über die Mismatch-Reparatur (MMR) unter Generierung von DNA-Doppelstrangbrüchen (DSBs). Die Untersuchung der durch Temozolomid induzierten DSBs, nachgewiesen durch gammaH2AX-Induktion, korrelierte direkt mit der apoptotischen Antwort von Melanomzelllinien und DSBs können somit als eine entscheidende apoptoseauslösende Größe angesehen werden. Eine Resistenz gegenüber dem methylierenden Temozolomid in der Zelllinie MZ7 konnte auf einen Defekt in der MMR-Schadenserkennung auf der Ebene des MutSalpha-Komplexes zurückgeführt werden. Dieser Defekt hatte keinen Einfluss auf die Fotemustin-vermittelte Apoptoseinduktion. Neben MGMT konnte somit die MMR als Resistenzfaktor gegenüber methylierenden Agenzien in Melanomen identifiziert werden. Die Fotemustin-induzierte Apoptose wurde in Melanomzelllinien im Detail untersucht. Es konnte erstmals gezeigt werden, dass Fotemustin-bedingte ICLs in Zellen einen G2/M-Arrest im Behandlungszyklus induzieren. Wie anhand G1-arretierter Zellen nachgewiesen werden konnte, war das Durchlaufen der DNA-Replikation vor Erreichen des Arrests für die Induktion der Apoptose notwendig. Die Prozessierung von ICLs ist im Vergleich zu Methylierungen der DNA deutlich komplexer. Dies könnte erklären, warum in Melanomzellen die durch gammaH2AX-Induktion repräsentierten DSBs nicht mit der Sensitivität der einzelnen Zelllinien korreliert. Die Untersuchung unterschiedlich sensitiver Zelllinien zeigte ein vergleichbares Schadensniveau an ICLs und eine ebenso vergleichbare initiale Prozessierung derselben unter Generierung von DSBs. Die Prozessierung dieser sekundären Läsionen, welche anhand der Abnahme von gammaH2AX-Foci untersucht wurde, war hingegen in der sensitiveren Melanomzelllinie deutlich weniger effektiv. Es konnte weiterhin nachgewiesen werden, dass eine uneffektive Prozessierung der sekundären Läsionen einhergeht mit einer verstärkten und länger anhaltenden Aktivierung der in der DSB-Detektion beteiligten Kinase ATM und der Checkpoint Kinase 1. Es wäre daher denkbar, dass eine verstärkte Aktivität dieser Kinasen proapoptotische Signale vermittelt. Unterschiede in der Prozessierung der sekundären Läsionen könnten somit ein wichtiger Marker der ICL-induzierten Apoptose darstellen. Des weitern konnte nachgewiesen werden, dass nach Fotemustingabe die mitochondrial-vermittelte Apoptose einen effektiven Exekutionsweg in Melanomen darstellt. Während Cytochrom C-Freigabe, Bcl-2-Abnahme an den Mitochondrien, Bax-Rekrutierung und Caspase-9 Aktivität nachgewiesen werden konnten, wurden keine Hinweise auf eine Fas-Rezeptor-vermittelte Apoptose gefunden. Die Unfähigkeit, Rezeptor-vermittelte Apoptose zu unterlaufen, könnte die Bedeutungslosigkeit des p53-Gens in Melanomen begründen, da gerade dieser Weg in der Alkylantien-induzierten Apoptose in anderen Zellsystemen eine große Relevanz besitzt. Bei der Suche nach einem alternativen proapoptotischen Signalweg konnten Hinweise für die Beteiligung des Rb/E2F-1-Wegs, welcher über p73 agiert, in einer p53-mutierten Melanomzelllinie gefunden werden. Einen Einfluss der Proteine Survivin und XIAP als Resistenzfaktoren auf die Fotemustin-induzierte Apoptose wurde hingegen nicht nachgewiesen.

High frequency acoustics in colloid-based meso- and nanostructures by spontaneous Brillouin light scattering

Materials that can mold the flow of elastic waves of certain energy in certain directions are called phononic materials. The present thesis deals essentially with such phononic systems, which are structured in the mesoscale (<1 µm), and with their individual components. Such systems show interesting phononic properties in the hypersonic region, i.e., at frequencies in the GHz range. It is shown that colloidal systems are excellent model systems for the realization of such phononic materials. Therefore, different structures and particle architectures are investigated by Brillouin light scattering, the inelastic scattering of light by phonons.rnThe experimental part of this work is divided into three chapters: Chapter 4 is concerned with the localized mechanical waves in the individual spherical colloidal particles, i.e., with their resonance- or eigenvibrations. The investigation of these vibrations with regard to the environment of the particles, their chemical composition, and the influence of temperature on nanoscopically structured colloids allows novel insights into the physical properties of colloids at small length scales. Furthermore, some general questions concerning light scattering on such systems, in dispute so far, are convincingly addressed.rnChapter 5 is a study of the traveling of mechanical waves in colloidal systems, consisting of ordered and disordered colloids in liquid or elastic matrix. Such systems show acoustic band gaps, which can be explained geometrically (Bragg gap) or by the interaction of the acoustic band with the eigenvibrations of the individual spheres (hybridization gap).rnWhile the latter has no analogue in photonics, the presence of strong phonon scatterers, when a large elastic mismatch between the composite components exists, can largely impact phonon propagation in analogy to strong multiple light scattering systems. The former is exemplified in silica based phononic structures that opens the door to new ways of sound propagation manipulation.rnChapter 6 describes the first measurement of the elastic moduli in newly fabricated by physical vapor deposition so-called ‘stable organic glasses’. rnIn brief, this thesis explores novel phenomena in colloid-based hypersonic phononic structures, utilizing a versatile microfabrication technique along with different colloid architectures provided by material science, and applying a non-destructive optical experimental tool to record dispersion diagrams.rn

Studio dei polimorfismi genici degli antigeni minori di istocompatibilità e GvHD/GvL nel trapianto allogenico di cellule staminali emopoietiche

L'outcome dei pazienti sottoposti a trapianto allogenico di cellule staminali emopoietiche è fortemente influenzato da graft versus leukemia (GvL) e graft versus host disease (GvHD) che sono mediate, almeno in parte, dagli antigeni minori di istocompatibilità (mHAgs). In letteratura sono stati identificati 26 mHAgs che sono stati correlati a GvHD/GvL con risultati incompleti e in alcuni casi contrastanti; inoltre manca una metodica che sia in grado di genotipizzare contemporaneamente un pannello così ampio. Il lavoro è stato finalizzato alla preparazione di un protocollo di laboratorio che permetta di studiare in modo efficace i 26 mHAgs identificati, per poi correlarli con GvHD/GvL all’interno di uno specifico gruppo di trapiantati. Utilizzando la metodica IPlex Gold Mass Array Sequenom e tecniche di biologia molecolare convenzionale sono stati genotipizzati 26 antigeni minori di istocompatibilità per 46 coppie full-matched. Tutti i pazienti inclusi nel progetto di studio erano stati sottoposti a trapianto allogenico di cellule staminali emopoietiche da donatore familiare o volontario full-compatibile per leucemia mieloide cronica (n=46) o leucemia acuta linfoblastica Philadelphia positiva (LAL-Ph+, n=24). Il progetto ha confermato l'efficienza (98.6%) e la fattibilità delle metodiche proposte. Dal lavoro è inoltre emerso che, le differenze tra donatore e ricevente a libello mHAgs ACC-1, ACC-4, ACC-5, LB-MTHFD1-1Q, UGT2B17, DPH1, LRH1 potrebbero essere fattori predittivi di GvHD (p<0.05). La seconda evidenza è legata a un trend secondo cui il mismatch per LB-ADIR1 protegge dalla recidiva di malattia, in particolare nei confronti della LAL-Ph+ che è scarsamente responsiva all'allo-immunoterapia. Questo lavoro pilota, la cui casistica deve quindi essere ampliata, ha dimostrato l’efficacia della genotipizzazione con IPlex Gold Sequenom e l’elevato potenziale degli mHAgs sia come fattori predittivi di GvHD che come driver di GvL.

Enhancing the efficiency of solar concentrators by controlled optical aberrations

The diameters of traditional dish concentrators can reach several tens of meters, the construction of monolithic mirrors being difficult at these scales: cheap flat reflecting facets mounted on a common frame generally reproduce a paraboloidal surface. When a standard imaging mirror is coupled with a PV dense array, problems arise since the solar image focused is intrinsically circular. Moreover, the corresponding irradiance distribution is bell-shaped in contrast with the requirement of having all the cells under the same illumination. Mismatch losses occur when interconnected cells experience different conditions, in particular in series connections. In this PhD Thesis, we aim at solving these issues by a multidisciplinary approach, exploiting optical concepts and applications developed specifically for astronomical use, where the improvement of the image quality is a very important issue. The strategy we propose is to boost the spot uniformity acting uniquely on the primary reflector and avoiding the big mirrors segmentation into numerous smaller elements that need to be accurately mounted and aligned. In the proposed method, the shape of the mirrors is analytically described by the Zernike polynomials and its optimization is numerically obtained to give a non-imaging optics able to produce a quasi-square spot, spatially uniform and with prescribed concentration level. The freeform primary optics leads to a substantial gain in efficiency without secondary optics. Simple electrical schemes for the receiver are also required. The concept has been investigated theoretically modeling an example of CPV dense array application, including the development of non-optical aspects as the design of the detector and of the supporting mechanics. For the method proposed and the specific CPV system described, a patent application has been filed in Italy with the number TO2014A000016. The patent has been developed thanks to the collaboration between the University of Bologna and INAF (National Institute for Astrophysics).

Nanoscale surface properties and interaction with fundamental biomolecules of chlorite and phlogopite

The surface properties of minerals have important implications in geology, environment, industry and biotechnology and for certain aspects in the research on the origin of life. This research project aims to widen the knowledge on the nanoscale surface properties of chlorite and phlogopite by means of advanced methodologies, and also to investigate the interaction of fundamental biomolecules, such as nucleotides, RNA, DNA and amino acid glycine with the surface of the selected phyllosilicates. Multiple advanced and complex experimental approaches based on scanning probe microscopy and spatially resolved spectroscopy were used and in some cases specifically developed. The results demonstrate that chlorite exposes at the surface atomically flat terraces with 0.5 nm steps typically generated by the fragmentation of the octahedral sheet of the interlayer (brucitic-type). This fragmentation at the nanoscale generates a high anisotropy and inhomogeneity with surface type and isomorphous cationic substitutions determining variations of the effective surface potential difference, ranging between 50-100 mV and 400-500 mV, when measured in air, between the TOT surface and the interlayer brucitic sheet. The surface potential was ascribed to be the driving force of the observed high affinity of the surface with the fundamental biomolecules, like single molecules of nucleotides, DNA, RNA and amino acids. Phlogopite was also observed to present an extended atomically flat surface, featuring negative surface potential values of some hundreds of millivolts and no significant local variations. Phlogopite surface was sometimes observed to present curvature features that may be ascribed to local substitutions of the interlayer cations or the presence of a crystal lattice mismatch or structural defects, such as stacking faults or dislocation loops. Surface chemistry was found similar to the bulk. The study of the interaction with nucleotides and glycine revealed a lower affinity with respect to the brucite-like surface of chlorite.

Educational mismathc in Italian an European labour markets (starting from the UK)

The present doctoral thesis is structured as a collection of three essays. The first essay, “SOC(HE)-Italy: a classification for graduate occupations” presents the conceptual basis, the construction, the validation and the application to the Italian labour force of the occupational classification termed SOC(HE)-Italy. I have developed this classification under the supervision of Kate Purcell during my period as a visiting research student at the Warwick Institute for Emplyment Research. This classification links the constituent tasks and duties of a particular job to the relevant knowledge and skills imparted via Higher Education (HE). It is based onto the SOC(HE)2010, an occupational classification first proposed by Kate Purcell in 2013, but differently constructed. In the second essay “Assessing the incidence and wage effects of overeducation among Italian graduates using a new measure for educational requirements” I utilize this classification to build a valid and reliable measure for job requirements. The lack of an unbiased measure for this dimension constitutes one of the major constraints to achieve a generally accepted measurement of overeducation. Estimations of overeducation incidence and wage effects are run onto AlmaLaurea data from the survey on graduates career paths. I have written this essay and obtained these estimates benefiting of the help and guidance of Giovanni Guidetti and Giulio Pedrini. The third and last essay titled “Overeducation in the Italian labour market: clarifying the concepts and addressing the measurement error problem” addresses a number of theoretical issues concerning the concepts of educational mismatch and overeducation. Using Istat data from RCFL survey I run estimates of the ORU model for the whole Italian labour force. In my knowledge, this is the first time ever such model is estimated on such population. In addition, I adopt the new measure of overeducation based onto the SOC(HE)-Italy classification.