New functional and biomechanical assessments for the improvement of the surgical navigation systems for joint replacement in the human lower limb

In case of severe osteoarthritis at the knee causing pain, deformity, and loss of stability and mobility, the clinicians consider that the substitution of these surfaces by means of joint prostheses. The objectives to be pursued by this surgery are: complete pain elimination, restoration of the normal physiological mobility and joint stability, correction of all deformities and, thus, of limping. The knee surgical navigation systems have bee developed in computer-aided surgery in order to improve the surgical final outcome in total knee arthroplasty. These systems provide the surgeon with quantitative and real-time information about each surgical action, like bone cut executions and prosthesis component alignment, by mean of tracking tools rigidly fixed onto the femur and the tibia. Nevertheless, there is still a margin of error due to the incorrect surgical procedures and to the still limited number of kinematic information provided by the current systems. Particularly, patello-femoral joint kinematics is not considered in knee surgical navigation. It is also unclear and, thus, a source of misunderstanding, what the most appropriate methodology is to study the patellar motion. In addition, also the knee ligamentous apparatus is superficially considered in navigated total knee arthroplasty, without taking into account how their physiological behavior is altered by this surgery. The aim of the present research work was to provide new functional and biomechanical assessments for the improvement of the surgical navigation systems for joint replacement in the human lower limb. This was mainly realized by means of the identification and development of new techniques that allow a thorough comprehension of the functioning of the knee joint, with particular attention to the patello-femoral joint and to the main knee soft tissues. A knee surgical navigation system with active markers was used in all research activities presented in this research work. Particularly, preliminary test were performed in order to assess the system accuracy and the robustness of a number of navigation procedures. Four studies were performed in-vivo on patients requiring total knee arthroplasty and randomly implanted by means of traditional and navigated procedures in order to check for the real efficacy of the latter with respect to the former. In order to cope with assessment of patello-femoral joint kinematics in the intact and replaced knees, twenty in-vitro tests were performed by using a prototypal tracking tool also for the patella. In addition to standard anatomical and articular recommendations, original proposals for defining the patellar anatomical-based reference frame and for studying the patello-femoral joint kinematics were reported and used in these tests. These definitions were applied to two further in-vitro tests in which, for the first time, also the implant of patellar component insert was fully navigated. In addition, an original technique to analyze the main knee soft tissues by means of anatomical-based fiber mappings was also reported and used in the same tests. The preliminary instrumental tests revealed a system accuracy within the millimeter and a good inter- and intra-observer repeatability in defining all anatomical reference frames. In in-vivo studies, the general alignments of femoral and tibial prosthesis components and of the lower limb mechanical axis, as measured on radiographs, was more satisfactory, i.e. within ±3°, in those patient in which total knee arthroplasty was performed by navigated procedures. As for in-vitro tests, consistent patello-femoral joint kinematic patterns were observed over specimens throughout the knee flexion arc. Generally, the physiological intact knee patellar motion was not restored after the implant. This restoration was successfully achieved in the two further tests where all component implants, included the patellar insert, were fully navigated, i.e. by means of intra-operative assessment of also patellar component positioning and general tibio-femoral and patello-femoral joint assessment. The tests for assessing the behavior of the main knee ligaments revealed the complexity of the latter and the different functional roles played by the several sub-bundles compounding each ligament. Also in this case, total knee arthroplasty altered the physiological behavior of these knee soft tissues. These results reveal in-vitro the relevance and the feasibility of the applications of new techniques for accurate knee soft tissues monitoring, patellar tracking assessment and navigated patellar resurfacing intra-operatively in the contest of the most modern operative techniques. This present research work gives a contribution to the much controversial knowledge on the normal and replaced of knee kinematics by testing the reported new methodologies. The consistence of these results provides fundamental information for the comprehension and improvements of knee orthopedic treatments. In the future, the reported new techniques can be safely applied in-vivo and also adopted in other joint replacements.

Transcriptional functions of DNA Topoisomerases at a genome-wide scale and a single gene levels

The DNA topology is an important modifier of DNA functions. Torsional stress is generated when right handed DNA is either over- or underwound, producing structural deformations which drive or are driven by processes such as replication, transcription, recombination and repair. DNA topoisomerases are molecular machines that regulate the topological state of the DNA in the cell. These enzymes accomplish this task by either passing one strand of the DNA through a break in the opposing strand or by passing a region of the duplex from the same or a different molecule through a double-stranded cut generated in the DNA. Because of their ability to cut one or two strands of DNA they are also target for some of the most successful anticancer drugs used in standard combination therapies of human cancers. An effective anticancer drug is Camptothecin (CPT) that specifically targets DNA topoisomerase 1 (TOP 1). The research project of the present thesis has been focused on the role of human TOP 1 during transcription and on the transcriptional consequences associated with TOP 1 inhibition by CPT in human cell lines. Previous findings demonstrate that TOP 1 inhibition by CPT perturbs RNA polymerase (RNAP II) density at promoters and along transcribed genes suggesting an involvement of TOP 1 in RNAP II promoter proximal pausing site. Within the transcription cycle, promoter pausing is a fundamental step the importance of which has been well established as a means of coupling elongation to RNA maturation. By measuring nascent RNA transcripts bound to chromatin, we demonstrated that TOP 1 inhibition by CPT can enhance RNAP II escape from promoter proximal pausing site of the human Hypoxia Inducible Factor 1 (HIF-1) and c-MYC genes in a dose dependent manner. This effect is dependent from Cdk7/Cdk9 activities since it can be reversed by the kinases inhibitor DRB. Since CPT affects RNAP II by promoting the hyperphosphorylation of its Rpb1 subunit the findings suggest that TOP 1inhibition by CPT may increase the activity of Cdks which in turn phosphorylate the Rpb1 subunit of RNAP II enhancing its escape from pausing. Interestingly, the transcriptional consequences of CPT induced topological stress are wider than expected. CPT increased co-transcriptional splicing of exon1 and 2 and markedly affected alternative splicing at exon 11. Surprisingly despite its well-established transcription inhibitory activity, CPT can trigger the production of a novel long RNA (5’aHIF-1) antisense to the human HIF-1 mRNA and a known antisense RNA at the 3’ end of the gene, while decreasing mRNA levels. The effects require TOP 1 and are independent from CPT induced DNA damage. Thus, when the supercoiling imbalance promoted by CPT occurs at promoter, it may trigger deregulation of the RNAP II pausing, increased chromatin accessibility and activation/derepression of antisense transcripts in a Cdks dependent manner. A changed balance of antisense transcripts and mRNAs may regulate the activity of HIF-1 and contribute to the control of tumor progression After focusing our TOP 1 investigations at a single gene level, we have extended the study to the whole genome by developing the “Topo-Seq” approach which generates a map of genome-wide distribution of sites of TOP 1 activity sites in human cells. The preliminary data revealed that TOP 1 preferentially localizes at intragenic regions and in particular at 5’ and 3’ ends of genes. Surprisingly upon TOP 1 downregulation, which impairs protein expression by 80%, TOP 1 molecules are mostly localized around 3’ ends of genes, thus suggesting that its activity is essential at these regions and can be compensate at 5’ ends. The developed procedure is a pioneer tool for the detection of TOP 1 cleavage sites across the genome and can open the way to further investigations of the enzyme roles in different nuclear processes.

Physical models for numerical simulation of Si-based nanoscale FETs and sensors

To continuously improve the performance of metal-oxide-semiconductor field-effect-transistors (MOSFETs), innovative device architectures, gate stack engineering and mobility enhancement techniques are under investigation. In this framework, new physics-based models for Technology Computer-Aided-Design (TCAD) simulation tools are needed to accurately predict the performance of upcoming nanoscale devices and to provide guidelines for their optimization. In this thesis, advanced physically-based mobility models for ultrathin body (UTB) devices with either planar or vertical architectures such as single-gate silicon-on-insulator (SOI) field-effect transistors (FETs), double-gate FETs, FinFETs and silicon nanowire FETs, integrating strain technology and high-κ gate stacks are presented. The effective mobility of the two-dimensional electron/hole gas in a UTB FETs channel is calculated taking into account its tensorial nature and the quantization effects. All the scattering events relevant for thin silicon films and for high-κ dielectrics and metal gates have been addressed and modeled for UTB FETs on differently oriented substrates. The effects of mechanical stress on (100) and (110) silicon band structures have been modeled for a generic stress configuration. Performance will also derive from heterogeneity, coming from the increasing diversity of functions integrated on complementary metal-oxide-semiconductor (CMOS) platforms. For example, new architectural concepts are of interest not only to extend the FET scaling process, but also to develop innovative sensor applications. Benefiting from properties like large surface-to-volume ratio and extreme sensitivity to surface modifications, silicon-nanowire-based sensors are gaining special attention in research. In this thesis, a comprehensive analysis of the physical effects playing a role in the detection of gas molecules is carried out by TCAD simulations combined with interface characterization techniques. The complex interaction of charge transport in silicon nanowires of different dimensions with interface trap states and remote charges is addressed to correctly reproduce experimental results of recently fabricated gas nanosensors.

Movement Analysis by means of inertial sensors: from bench to bedside

in the everyday clinical practice. Having this in mind, the choice of a simple setup would not be enough because, even if the setup is quick and simple, the instrumental assessment would still be in addition to the daily routine. The will to overcome this limit has led to the idea of instrumenting already existing and widely used functional tests. In this way the sensor based assessment becomes an integral part of the clinical assessment. Reliable and validated signal processing methods have been successfully implemented in Personal Health Systems based on smartphone technology. At the end of this research project there is evidence that such solution can really and easily used in clinical practice in both supervised and unsupervised settings. Smartphone based solution, together or in place of dedicated wearable sensing units, can truly become a pervasive and low-cost means for providing suitable testing solutions for quantitative movement analysis with a clear clinical value, ultimately providing enhanced balance and mobility support to an aging population.

Celestial Church of Christ in Italia. Percorsi, Spazi e Pratiche Rituali

Il lavoro che presento propone un’analisi di una chiesa africana indipendente in Italia, la Celestial Church Of Christ Worldwide (CCCW), cercando di mettere in luce il nesso tra religione, migrazione e il processo di ‘plunting churches’ (Kooning 2009) nel contesto italiano. Attraverso una ricerca sul campo, sono stati indagati i percorsi personali, familiari e comunitari dei membri di una ‘Celestial Parish’ presente nel comune di Brescia, ‘Ileri Oluwa Parish’, al fine di comprendere la natura dei processi identitari coinvolti nell’organizzazione della CCC in Italia. ‘Ileri Oluwa Parish’, in quanto luogo che denota una ‘chiesa individuale collegata ad una Diocesi’ (CCC Constitution (CCC Constitution, 107 (d) si rivela, nella materialità delle sue forme e dei ‘Devotional Services’ che in essa si svolgono, a ‘field of action’ (Lefebvre, 1991). La storia della chiesa, i fondamenti della sua dottrina e i significati comunicati attraverso le forme rituali e religiose che la stessa promuove, sono stati contestualizzati alla luce delle tensioni e delle strategie di potere che strutturano il campo. Le storie dei membri della parrocchia, percorsi di migrazione e mobilità in itinere, rappresentano la lente attraverso cui si è guardato alle relazioni vissute nel nome dello ‘Spirito’, e alla percezione stessa di ciò che gli stessi Celestians definiscono sacro, santo, puro e impuro. Lo sguardo fisso alla vita ordinaria di una Celestial parish in Italia, esteso nell’ultima parte dell’elaborato alla Celestial parish londinese, è stato fondamentale per capire l’intreccio di relazioni spirituali, reti familiari e mobilità degli individui sul territorio italiano ed europeo, processo che ribalta la condizione diasporica della CCC, trasformando una condizione di dispersione in un valore aggiunto, nella possibilità di nuove traiettorie territoriali e spazi di presenza religiosa e socioeconomica.

Il cambiamento della ruralità nell'Unione Europea. Tipologie, evoluzione e risposte alle politiche

Obiettivo del lavoro è migliorare la lettura della ruralità europea. A fronte delle profonde trasformazioni avvenute, oggi non è più possibile analizzare i territori rurali adottando un mero approccio dicotomico che semplicemente li distingua dalle città. Al contrario, il lavoro integra l’analisi degli aspetti socio-economici con quella degli elementi territoriali, esaltando le principali dimensioni che caratterizzano le tante tipologie di ruralità oggi presenti in Europa. Muovendo dal dibattito sulla classificazione delle aree rurali, si propone dapprima un indicatore sintetico di ruralità che, adottando la logica fuzzy, considera congiuntamente aspetti demografici (densità), settoriali (rilevanza dell’attività agricola), territoriali e geografici (accessibilità e uso del suolo). Tale tecnica permette di ricostruire un continuum di gradi di ruralità, distinguendo così, all’interno dell’Unione Europea (circa 1.300 osservazioni), le aree più centrali da quelle progressivamente più rurali e periferiche. Successivamente, attraverso un’analisi cluster vengono individuate tipologie di aree omogenee in termini di struttura economica, paesaggio, diversificazione dell’attività agricola. Tali cluster risentono anche della distribuzione geografica delle aree stesse: vengono infatti distinti gruppi di regioni centrali da gruppi di regioni più periferiche. Tale analisi evidenzia soprattutto come il binomio ruralità-arretratezza risulti ormai superato: alcune aree rurali, infatti, hanno tratto vantaggio dalle trasformazioni che hanno interessato l’Unione Europea negli ultimi decenni (diffusione dell’ICT o sviluppo della manifattura). L’ultima parte del lavoro offre strumenti di analisi a supporto dell’azione politica comunitaria, analizzando la diversa capacità delle regioni europee di rispondere alle sfide lanciate dalla Strategia Europa 2020. Un’analisi in componenti principali sintetizza le principali dimensioni di tale performance regionale: i risultati sono poi riletti alla luce delle caratteristiche strutturali dei territori europei. Infine, una più diretta analisi spaziale dei dati permette di evidenziare come la geografia influenzi ancora profondamente la capacità dei territori di rispondere alle nuove sfide del decennio.