Rôle du gène de fusion MLL-ENL dans le développement et le maintien du phénotype leucémique

Les translocations chromosomiques du gène MLL sont connues pour mener au développement de leucémies aiguës. La translocation avec un de ses partenaires de fusion les plus communs, ENL, peut engendrer des leucémies aiguës de plusieurs types différents pour cette même translocation. Une fois la leucémogenèse initiée par la fusion MLL-ENL, son rôle quant au maintien du phénotype leucémique n’est pas encore bien connu à ce jour. Pour mieux comprendre l’importance de MLL-ENL après la leucémogenèse, des cellules souches/progénitrices de sang de cordon ombilical humain purifiées ont ainsi été transduites par un virus exprimant le gène de fusion MLL-ENL bordé par des sites LoxP ainsi que le marqueur eGFP. Ces cellules infectées ont ensuite été injectées dans notre modèle de souris immunodéficientes irradiées et placées sous observation pendant 24 semaines pour voir le développement de leucémies aiguës. Elles ont alors été sacrifiées et les cellules la moelle osseuse et de la rate ont ensuite été analysées par cytométrie en flux pour déterminer si la xénogreffe a engendré une leucémie dans notre modèle ainsi que le phénotype de celle-ci. Les souris injectées par les cellules infectées par le MLL-ENL ont généré uniquement des leucémies lymphoïdes aiguës de type B. Les cellules de ces leucémies primaires isolées ont été par la suite infectées par un lentivirus exprimant la cre-recombinase et le marqueur BFP afin d’exciser le gène MLL-ENL des cellules leucémiques grâce aux sites LoxP. Les cellules ont ensuite été triées pour le marqueur BFP et injectées dans des souris secondaires pour de voir si les cellules leucémiques souches pouvaient toujours régénérer la leucémie. Les conséquences de l’absence de MLL-ENL dans le maintien du phénotype leucémique n’ont cependant pas pu être vérifiées à cause d’une erreur dans la séquence de la cre-recombinase, mais nous avons observé la régénération des leucémies secondaires.

Development of Core Design Methods and Tools for Gen-IV Heavy Liquid Metal Cooled Reactors

Heavy Liquid Metal Cooled Reactors are among the concepts, fostered by the GIF, as potentially able to comply with stringent safety, economical, sustainability, proliferation resistance and physical protection requirements. The increasing interest around these innovative systems has highlighted the lack of tools specifically dedicated to their core design stage. The present PhD thesis summarizes the three years effort of, partially, closing the mentioned gap, by rationally defining the role of codes in core design and by creating a development methodology for core design-oriented codes (DOCs) and its subsequent application to the most needed design areas. The covered fields are, in particular, the fuel assembly thermal-hydraulics and the fuel pin thermo-mechanics. Regarding the former, following the established methodology, the sub-channel code ANTEO+ has been conceived. Initially restricted to the forced convection regime and subsequently extended to the mixed one, ANTEO+, via a thorough validation campaign, has been demonstrated a reliable tool for design applications. Concerning the fuel pin thermo-mechanics, the will to include safety-related considerations at the outset of the pin dimensioning process, has given birth to the safety-informed DOC TEMIDE. The proposed DOC development methodology has also been applied to TEMIDE; given the complex interdependence patterns among the numerous phenomena involved in an irradiated fuel pin, to optimize the code final structure, a sensitivity analysis has been performed, in the anticipated application domain. The development methodology has also been tested in the verification and validation phases; the latter, due to the low availability of experiments truly representative of TEMIDE's application domain, has only been a preliminary attempt to test TEMIDE's capabilities in fulfilling the DOC requirements upon which it has been built. In general, the capability of the proposed development methodology for DOCs in delivering tools helping the core designer in preliminary setting the system configuration has been proven.

Inertial and magnetic sensors for upper limb kinematics: modeling, calibration, sensor fusion and clinical applications

The primary aim of the research activity presented in this PhD thesis was the development of an innovative hardware and software solution for creating a unique tool for kinematics and electromyographic analysis of the human body in an ecological setting. For this purpose, innovative algorithms have been proposed regarding different aspects of inertial and magnetic data elaboration: magnetometer calibration and magnetic field mapping (Chapter 2), data calibration (Chapter 3) and sensor-fusion algorithm. Topics that may conflict with the confidentiality agreement between University of Bologna and NCS Lab will not be covered in this thesis. After developing and testing the wireless platform, research activities were focused on its clinical validation. The first clinical study aimed to evaluate the intra and interobserver reproducibility in order to evaluate three-dimensional humero-scapulo-thoracic kinematics in an outpatient setting (Chapter 4). A second study aimed to evaluate the effect of Latissimus Dorsi Tendon Transfer on shoulder kinematics and Latissimus Dorsi activation in humerus intra - extra rotations (Chapter 5). Results from both clinical studies have demonstrated the ability of the developed platform to enter into daily clinical practice, providing useful information for patients' rehabilitation.

Hydrogen production by enhanced methane reforming with membrane reactors or with CO2 capture materials

Pure hydrogen production from methane is a multi-step process run on a large scale for economic reasons. However, hydrogen can be produced in a one-pot continuous process for small scale applications, namely Low Temperature Steam Reforming. Here, Steam Reforming is carried out in a reactor whose walls are composed by a membrane selective toward hydrogen. Pd is the most used membrane material due to its high permeability and selectivity. However, Pd deteriorates at temperatures higher than 500°C, thus the operative temperature of the reaction has to be lowered. However, the employment of a membrane reactor may allow to give high yields thanks to hydrogen removal, which shifts the reaction toward the products. Moreover, pure hydrogen is produced. This work is concentrated on the synthesis of a catalytic system and the investigation of its performances in different processes, namely oxy-reforming, steam reforming and water gas shift, to find appropriate conditions for hydrogen production in a catalytic membrane reactor. The catalyst supports were CeZr and Zr oxides synthesized by microemulsion, impregnated with different noble metals. Pt, Rh and PtRh based catalysts were tested in the oxy reforming process at 500°C, where Rh on CeZr gave the most interesting results. On the opposite, the best performances in low temperature steam reforming were obtained with Rh impregnated on Zr oxide. This catalyst was selected to perform low temperature steam reforming in a Pd membrane reactor. The hydrogen removal given by the membrane allowed to increase the methane conversion over the equilibrium of a classical fixed bed reactor thanks to an equilibrium shift effect. High hydrogen production and recoveries were also obtained, and no other compound permeated through the membrane which proved to be hydrogen selective.

Optimization of an innovative T6 heat treatment for the AlSi10Mg alloy processed by Laser-based Powder Bed Fusion: effect on microstructure, mechanical properties, and tribological behavior

Laser-based Powder Bed Fusion (L-PBF) technology is one of the most commonly used metal Additive Manufacturing (AM) techniques to produce highly customized and value-added parts. The AlSi10Mg alloy has received more attention in the L-PBF process due to its good printability, high strength/weight ratio, corrosion resistance, and relatively low cost. However, a deep understanding of the effect of heat treatments on this alloy's metastable microstructure is still required for developing tailored heat treatments for the L-PBF AlSi10Mg alloy to overcome the limits of the as-built condition. Several authors have already investigated the effects of conventional heat treatment on the microstructure and mechanical behavior of the L-PBF AlSi10Mg alloy but often overlooked the peculiarities of the starting supersatured and ultrafine microstructure induced by rapid solidification. For this reason, the effects of innovative T6 heat treatment (T6R) on the microstructure and mechanical behavior of the L-PBF AlSi10Mg alloy were assessed. The short solution soaking time (10 min) and the relatively low temperature (510 °C) reduced the typical porosity growth at high temperatures and led to a homogeneous distribution of fine globular Si particles in the Al matrix. In addition, it increased the amount of Mg and Si in the solid solution available for precipitation hardening during the aging step. The mechanical (at room temperature and 200 °C) and tribological properties of the T6R alloy were evaluated and compared with other solutions, especially with an optimized direct-aged alloy (T5 alloy). Results showed that the innovative T6R alloy exhibits the best mechanical trade-off between strength and ductility, the highest fatigue strength among the analyzed conditions, and interesting tribological behavior. Furthermore, the high-temperature mechanical performances of the heat-treated L-PBF AlSi10Mg alloy make it suitable for structural components operating in mild service conditions at 200 °C.

High-strength steels manufactured via Laser Powder Bed Fusion: effect of post-process heat treatments and surface finishing on microstructure and mechanical properties

Laser Powder Bed Fusion (LPBF) permits the manufacturing of parts with optimized geometry, enabling lightweight design of mechanical components in aerospace and automotive and the production of tools with conformal cooling channels. In order to produce parts with high strength-to-weight ratio, high-strength steels are required. To date, the most diffused high-strength steels for LPBF are hot-work tool steels, maraging and precipitation-hardening stainless steels, featuring different composition, feasibility and properties. Moreover, LPBF parts usually require a proper heat treatment and surface finishing, to develop the desired properties and reduce the high roughness resulting from LPBF. The present PhD thesis investigates the effect of different heat treatments and surface finishing on the microstructure and mechanical properties of a hot-work tool steel and a precipitation-hardening stainless steel manufactured via LPBF. The bibliographic section focuses on the main aspects of LPBF, hot-work tool steels and precipitation-hardening stainless steels. The experimental section is divided in two parts. Part A addresses the effect of different heat treatments and surface finishing on the microstructure, hardness, tensile and fatigue behaviour of a LPBF manufactured hot-work tool steel, to evaluate its feasibility for automotive and racing components. Results indicated the possibility to achieve high hardness and strength, comparable to the conventionally produced steel, but a great sensitivity of fatigue strength on defects and surface roughness resulting from LPBF. Part B investigates the effect of different heat treatments on the microstructure, hardness, tensile and notch-impact behaviour of a LPBF produced precipitation-hardening stainless steel, to assess its feasibility for tooling applications. Results indicated the possibility to achieve high hardness and strength also through a simple Direct Aging, enabling heat treatment simplification by exploiting the microstructural features resulting from LPBF.

Influenza dei difetti sul comportamento a fatica della lega AlSi10Mg prodotta con Laser Powder Bed Fusion

La tesi in oggetto ha lo scopo di valutare la fattibilità di sviluppare, sulla base di dati sperimentali, un metodo previsionale per il comportamento a fatica della lega AlSi10Mg prodotta mediante Laser Powder Bed Fusion, adattabile a componenti di dimensioni qualsiasi. La lega è stata trattata con invecchiamento diretto (T5) ottimizzato per ottenere una parziale riduzione delle tensioni residue senza comprometterne la durezza. Partendo da prove di fatica a flessione rotante condotte seguendo il metodo statistico Stair-Case per la definizione della resistenza a fatica, si vuole correlare il difetto killer, ossia quello che innesca la rottura del provino, alla caratterizzazione dei difetti condotta pre-prova con tomografia computerizzata ad alta risoluzione (µCT). Il difetto killer, osservato sulle superfici di frattura, viene analizzato in termini di dimensione, posizione e forma. Contemporaneamente è stato sviluppato un metodo per l’analisi dei difetti ottenuta con µCT tale da poter identificare quelli considerati più pericolosi, sulla base dello stato tensionale del campione, di posizione e dimensione dei difetti. Inoltre, è stata eseguita una caratterizzazione microstrutturale sulle sezioni trasversali dei campioni per valutare la quantità, dimensione e forma dei difetti e confrontarla con i dati di µCT. Per mostrare la relazione tra le dimensioni del difetto killer e la resistenza a fatica è stato adottato il diagramma di Kitagawa-Takahashi, con il modello di El Haddad. Dal confronto tra le metodologie è stata riscontrata una buona correlazione tra i difetti individuati come pericolosi dai dati di µCT e quelli che hanno effettivamente portato a rottura il campione, osservati in frattografia, supportando quindi la possibilità di sviluppare un metodo previsionale del comportamento a fatica. Quantitativamente, le dimensioni dei difetti killer sono nell’intervallo 90-130 µm e si trovano ad una distanza massima dalla superficie di non oltre 250 µm.

Dataset Fusion application on deep learning Object Detection task

Application of dataset fusion techniques to an object detection task, involving the use of deep learning as convolutional neural networks, to manage to create a single RCNN architecture able to inference with good performances on two distinct datasets with different domains.

Aspetti microstrutturali, proprietà meccaniche e ottimizzazione del trattamento termico di un acciaio inossidabile PH processato con laser powder bed fusion

L’attività di ricerca svolta in questa tesi ha riguardato gli aspetti microstrutturali, le proprietà meccaniche e l’ottimizzazione del trattamento termico di un acciaio inossidabile indurente per precipitazione processato mediante tecnologia additiva Laser Powder Bed Fusion (L-PBF). I provini, realizzati presso il competence center Bi-REX (Bologna) e lavorati presso il laboratorio di Metallurgia del DIN, sono stati oggetto di vari trattamenti termici sperimentali in cui è stata fatta variare la temperatura e la durata di mantenimento in forno, al fine di identificare la combinazione ottimale di temperatura e tempo sia per la fase di solubilizzazione che di invecchiamento. Nello specifico, la scelta dei parametri di solubilizzazione e invecchiamento è stata fatta solo sulla base dei trend di durezza. Da queste attività sono stati scelti i parametri di trattamento che garantissero elevata durezza, prossima alla massima ottenibile, senza durate di invecchiamento eccessive. L'esito della sperimentazione ha permesso di ridurre sensibilmente temperatura e durata di solubilizzazione rispetto al trattamento benchmark senza penalizzazione della durezza risultante. Dopodiché è stata eseguita una caratterizzazione microstrutturale e meccanica dell’acciaio, in termini di durezza, trazione e resilienza, per confrontare il trattamento termico ottimizzato con la condizione As-Built (AB) e con il trattamento termico standard indicato dal produttore. L’elevata durezza misurata dopo invecchiamento si deve alla presenza di precipitati di rinforzo in grado di ostacolare il moto delle dislocazioni. Le prove meccaniche hanno confermato l’ottenimento di elevata resistenza a snervamento e a trazione, superiore al benchmark, ma di un basso allungamento a rottura e bassissima resilienza a causa di difetti quali porosità, mancate fusioni e spattering. Infine, lo studio delle superfici di frattura ha permesso di analizzare i meccanismi di rottura dei campioni di trazione e resilienza.

Ossidazione anodica al plasma della lega di alluminio A357 prodotta mediante Laser - Powder Bed Fusion: microstruttura e comportamento tribologico

In questo elaborato è stato analizzato il comportamento tribologico della lega di alluminio A357 realizzata tramite Laser Powder Bed Fusion (L-PBF). In particolare, è stato studiato l’effetto del processo di anodizzazione ECO (ElectroChemical Oxidation) su tale lega, sia allo stato as-built che dopo trattamento termico T6R, in termini di attrito e usura a temperatura ambiente (i.e. 25°C) e a 200°C. Lo studio tribologico è stato svolto mediante prove di strisciamento non lubrificato in moto reciprocante (geometria ball-on-disc contro allumina). Nella prima parte del lavoro sono state effettuate prove a temperatura ambiente, su strati ECO a rugosità variabile, con carichi applicati nell’intervallo 1-8N. Nella seconda parte l’attività sperimentale si è invece rivolta al confronto nel comportamento tribologico della lega a 25°C e 200°C, mantenendo il carico invariato pari a 1 N e analizzando la risposta in termini di attrito e usura, sia per campioni rivestiti ECO che per campioni non rivestiti. Il rivestimento anodico ha mostrato a temperatura ambiente una maggiore resistenza ad usura quando applicato su superfici pre-lucidate, nonostante un maggiore coefficiente di attrito, dando luogo a cedimento completo ad un carico di 8 N (contro i 5 N del non lucidato). Dalla seconda fase di questo studio è invece risultato come il rivestimento ECO abbia notevolmente migliorato la resistenza ad usura rispetto al materiale non rivestito, sia a 25°C che a 200°C, mantenendo invariato il coefficiente d’attrito all’aumentare della temperatura, al contrario dei campioni A357 non rivestiti, che hanno invece manifestato un aumento del coefficiente di attrito di circa il doppio. Vale inoltre la pena notare come la pre-lucidatura ed il trattamento termico dei campioni A357 ECO siano risultati superflui in termini di comportamento tribologico; i campioni a superficie rugosa hanno dato luogo addirittura a minori coefficienti di attrito sia a 25°C che a 200°C, a parità di profondità d’usura.

Studio del comportamento meccanico e tribologico della lega AlSi10Mg prodotta mediante Laser-based Powder Bed Fusion sottoposta ad un ciclo tecnologico ottimizzato

AlSi10Mg alloy is one of the most widely used alloys for producing structural components by Laser-based Powder Fusion (L-PBF) technology due to the high mechanical and technological properties. The present work aims to characterize mechanically and tribologically the L-PBF AlSi10Mg alloy subjected to both heat treatment and surface modification cycles. Specifically, the effects of three heat treatments on the tribological and mechanical properties of the alloy were analyzed: T5 (artificial aging at 160 °C for 4 h), T6 rapid solution heat treatment (solution heat treatment at 510 °C for 1h and aging at 160 °C for 6 h), and T6 benchmark (solution heat treatment at 540 °C for 1h and aging at 160 °C for 4 h), the latter used as a benchmark. The study highlighted how the better balance between strength and ductility properties induced by the introduction of heat treatments leads to lower wear resistance and not significant variations in the friction coefficient of the alloy. The tribological and mechanical behavior of the alloy coated with two different coating structures, consisting of (i) chemical Ni (Ni-P) and (ii) Ni-P + DLC, was also evaluated. The goal was the identification of a deposition cycle such as to guarantee the optimization of the mechanical and tribological behavior of the alloy. The Ni-P coating provided good wear resistance but an increase in the coefficient of friction. In contrast, using the DLC top coating resulted in excellent tribological performance in wear resistance and friction coefficient. The samples characterized by the Ni-P + DLC multilayer coating were subsequently subjected to mechanical characterization. The results obtained highlighted problems of adhesion and incipient breaking of the material due to the different mechanical behavior of the coating, considerably reducing the mechanical performance of the alloy coated with Ni-P+DLC multilayer solution compared to the specimens in the un-coated condition.

Effetto della sabbiatura sul comportamento a fatica della lega AlSi10Mg prodotta con Laser Powder Bed Fusion e trattata termicamente

La tesi in oggetto ha lo scopo di determinare l’effetto della sabbiatura sul comportamento a fatica della lega AlSi10Mg prodotta mediante Laser Powder Bed Fusion e trattata termicamente. I parametri di processo e di trattamento termico (T5 e T6) sono stati precedentemente ottimizzati. Al fine di determinare l’effetto della sabbiatura su topografia superficiale e microstruttura dei campioni, si sono condotte molteplici analisi avvalendosi di strumenti quali profilometria, microscopia ottica ed in scansione, analisi di tensioni residue con diffrazione a raggi X e prove di durezza. Attraverso prove di fatica per flessione rotante, eseguite secondo il metodo Stair-Case per la determinazione della resistenza a fatica, e successiva caratterizzazione delle superfici di frattura si vuole correlare il difetto killer, ossia quello responsabile del cedimento per fatica, alle caratteristiche morfologiche e microstrutturali. Il difetto killer viene caratterizzato in termini di dimensione e distanza dalla superficie e per mostrare la relazione fra la dimensione del difetto killer e la resistenza a fatica si adotta il diagramma di Kitagawa-Takahashi con modellazione di Murakami ed EL Haddad. Si è evidenziato che tutti i difetti killer sono riconducibili a lack-of-fusion con dimensione superiore ai 100 μm ad una profondità compresa fra i 150 e i 200 μm, indipendentemente dal trattamento termico o meccanico applicato. In termini di fatica si osserva che il trattamento T6 conferisce al materiale migliori proprietà rispetto a quello T5. Il processo di sabbiatura, confrontato con quello di lucidatura superficiale, ha portato a miglioramenti in termini di durezza e tensioni residue di compressione, ma si è rivelato quasi ininfluente sulla resistenza a fatica. Sulla base di quanto sopra, si conferma la possibilità di applicazione della sabbiatura in ambito industriale a componenti meccanici, anche in sostituzione della lucidatura, ottenendo un beneficio anche economico.

Facial Changes After Early Treatment Of Unilateral Coronal Synostosis Question The Necessity Of Primary Nasal Osteotomy.

The premature fusion of unilateral coronal suture can cause a significant asymmetry of the craniofacial skeleton, with an oblique deviation of the cranial base that negatively impacts soft tissue facial symmetry. The purpose of this study was to assess facial symmetry obtained in patients with unilateral coronal synostosis (UCS) surgically treated by 2 different techniques. We hypothesized that nasal deviation should not be addressed in a primary surgical correction of UCS. Consecutive UCS patients were enrolled in a prospective study and randomly divided into 2 groups. In group 1, the patients underwent total frontal reconstruction and transferring of onlay bone grafts to the recessive superior orbital rim (n = 7), and in group 2, the patients underwent total frontal reconstruction and unilateral fronto-orbital advancement (n = 5). Computerized photogrammetric analysis measured vertical and horizontal axis of the nose and the orbital globe in the preoperative and postoperative periods. Intragroup and intergroup comparisons were performed. Intragroup preoperative and postoperative comparisons showed a significant (all P < 0.05) reduction of the nasal axis and the orbital-globe axis in the postoperative period in the 2 groups. Intergroup comparisons showed no significant difference (all P > 0.05). Facial symmetry was achieved in the patients with UCS who underwent surgery regardless of surgical approach evaluated here. Our data showed a significant improvement in nasal and orbital-globe deviation, leading us to question the necessity of primary nasal correction in these patients.

Avaliação da biodegradabilidade anaeróbia de resíduos da bovinocultura e da suinocultura

The biodegradability of animal wastes production was evaluated through a simplified methodology that allowed the verification of the applicability of anaerobic processes. The experiments were performed in bath reactors, with granular sludge of three origins: UASB reactor treating dairy effluent, UASB reactor treating swine effluent and UASB reactor treating effluent of slaughterhouse of poultry. The experiments (1) - dairy effluent and poultry slaughterhouse non-adapted sludge; (2) -swine effluent and poultry slaughterhouse non-adapted sludge; (3) - dairy effluent and poultry slaughterhouse adapted sludge; (4) - swine effluent and poultry slaughterhouse adapted sludge; (5) - dairy effluent and dairy sludge, and (6) - swine effluent and swine sludge were performed in Incubator Shaker, at a temperature of 35 °C, under agitation at a 150 rpm, for 5 minutes, every 1 hour. A substrat:biomass relationship of 0.5 was used. Kinetic models of Monod, Zero Order, First and Second Order were tested and it was verified that the First Order model provided the best adjustment. The apparent First Order kinetic parameter (k1) was estimated for the experiments 1; 2; 3; 4; 5, and 6, as 2.51 x 10-2; 2.49 x 10-2; 1.90 x 10-2; 3.09 x 10-2; 2.54 x 10-2; 4.09 x 10-2 h-1, respectively.

Flicker perimetry in healthy subjects: influence of age and gender, learning effect and short-term fluctuation

PURPOSE: To determine the mean critical fusion frequency and the short-term fluctuation, to analyze the influence of age, gender, and the learning effect in healthy subjects undergoing flicker perimetry. METHODS: Study 1 - 95 healthy subjects underwent flicker perimetry once in one eye. Mean critical fusion frequency values were compared between genders, and the influence of age was evaluated using linear regression analysis. Study 2 - 20 healthy subjects underwent flicker perimetry 5 times in one eye. The first 3 sessions were separated by an interval of 1 to 30 days, whereas the last 3 sessions were performed within the same day. The first 3 sessions were used to investigate the presence of a learning effect, whereas the last 3 tests were used to calculate short-term fluctuation. RESULTS: Study 1 - Linear regression analysis demonstrated that mean global, foveal, central, and critical fusion frequency per quadrant significantly decreased with age (p<0.05).There were no statistically significant differences in mean critical fusion frequency values between males and females (p>0.05), with the exception of the central area and inferonasal quadrant (p=0.049 and p=0.011, respectively), where the values were lower in females. Study 2 - Mean global (p=0.014), central (p=0.008), and peripheral (p=0.03) critical fusion frequency were significantly lower in the first session compared to the second and third sessions. The mean global short-term fluctuation was 5.06±1.13 Hz, the mean interindividual and intraindividual variabilities were 11.2±2.8% and 6.4±1.5%, respectively. CONCLUSION: This study suggests that, in healthy subjects, critical fusion frequency decreases with age, that flicker perimetry is associated with a learning effect, and that a moderately high short-term fluctuation is expected.