496 resultados para RSA
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Let G be a graph on n vertices with maximum degree ?. We use the Lovasz local lemma to show the following two results about colourings ? of the edges of the complete graph Kn. If for each vertex v of Kn the colouring ? assigns each colour to at most (n - 2)/(22.4?2) edges emanating from v, then there is a copy of G in Kn which is properly edge-coloured by ?. This improves on a result of Alon, Jiang, Miller, and Pritikin [Random Struct. Algorithms 23(4), 409433, 2003]. On the other hand, if ? assigns each colour to at most n/(51?2) edges of Kn, then there is a copy of G in Kn such that each edge of G receives a different colour from ?. This proves a conjecture of Frieze and Krivelevich [Electron. J. Comb. 15(1), R59, 2008]. Our proofs rely on a framework developed by Lu and Szekely [Electron. J. Comb. 14(1), R63, 2007] for applying the local lemma to random injections. In order to improve the constants in our results we use a version of the local lemma due to Bissacot, Fernandez, Procacci, and Scoppola [preprint, arXiv:0910.1824]. (c) 2011 Wiley Periodicals, Inc. Random Struct. Alg., 40, 425436, 2012
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We study quasi-random properties of k-uniform hypergraphs. Our central notion is uniform edge distribution with respect to large vertex sets. We will find several equivalent characterisations of this property and our work can be viewed as an extension of the well known Chung-Graham-Wilson theorem for quasi-random graphs. Moreover, let K(k) be the complete graph on k vertices and M(k) the line graph of the graph of the k-dimensional hypercube. We will show that the pair of graphs (K(k),M(k)) has the property that if the number of copies of both K(k) and M(k) in another graph G are as expected in the random graph of density d, then G is quasi-random (in the sense of the Chung-Graham-Wilson theorem) with density close to d. (C) 2011 Wiley Periodicals, Inc. Random Struct. Alg., 40, 1-38, 2012
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In this study we investigate the singlet excited state absorption of lutetium bisphthalocyanine (LuPc2) over a wide spectral range. It was observed distinct nonlinear absorption behaviors; saturable (SA) and reverse saturable absorption (RSA). The RSA effect was observed below 640 and above 680 nm, while SA occurs around the Q-band region, located around 660 nm. To describe the main singlet-singlet transitions, we employed the rate equation model considering the simplified three-energy level diagram. Our results reveal a ratio between excited and ground state absorption smaller than 0.05 at the Q-band region, and of approximately 4 for the other regions. (C) 2012 Elsevier B.V. All rights reserved.
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Abstract Background Sugarcane is an increasingly economically and environmentally important C4 grass, used for the production of sugar and bioethanol, a low-carbon emission fuel. Sugarcane originated from crosses of Saccharum species and is noted for its unique capacity to accumulate high amounts of sucrose in its stems. Environmental stresses limit enormously sugarcane productivity worldwide. To investigate transcriptome changes in response to environmental inputs that alter yield we used cDNA microarrays to profile expression of 1,545 genes in plants submitted to drought, phosphate starvation, herbivory and N2-fixing endophytic bacteria. We also investigated the response to phytohormones (abscisic acid and methyl jasmonate). The arrayed elements correspond mostly to genes involved in signal transduction, hormone biosynthesis, transcription factors, novel genes and genes corresponding to unknown proteins. Results Adopting an outliers searching method 179 genes with strikingly different expression levels were identified as differentially expressed in at least one of the treatments analysed. Self Organizing Maps were used to cluster the expression profiles of 695 genes that showed a highly correlated expression pattern among replicates. The expression data for 22 genes was evaluated for 36 experimental data points by quantitative RT-PCR indicating a validation rate of 80.5% using three biological experimental replicates. The SUCAST Database was created that provides public access to the data described in this work, linked to tissue expression profiling and the SUCAST gene category and sequence analysis. The SUCAST database also includes a categorization of the sugarcane kinome based on a phylogenetic grouping that included 182 undefined kinases. Conclusion An extensive study on the sugarcane transcriptome was performed. Sugarcane genes responsive to phytohormones and to challenges sugarcane commonly deals with in the field were identified. Additionally, the protein kinases were annotated based on a phylogenetic approach. The experimental design and statistical analysis applied proved robust to unravel genes associated with a diverse array of conditions attributing novel functions to previously unknown or undefined genes. The data consolidated in the SUCAST database resource can guide further studies and be useful for the development of improved sugarcane varieties.
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Primary stability of stems in cementless total hip replacements is recognized to play a critical role for long-term survival and thus for the success of the overall surgical procedure. In Literature, several studies addressed this important issue. Different approaches have been explored aiming to evaluate the extent of stability achieved during surgery. Some of these are in-vitro protocols while other tools are coinceived for the post-operative assessment of prosthesis migration relative to the host bone. In vitro protocols reported in the literature are not exportable to the operating room. Anyway most of them show a good overall accuracy. The RSA, EBRA and the radiographic analysis are currently used to check the healing process of the implanted femur at different follow-ups, evaluating implant migration, occurance of bone resorption or osteolysis at the interface. These methods are important for follow up and clinical study but do not assist the surgeon during implantation. At the time I started my Ph.D Study in Bioengineering, only one study had been undertaken to measure stability intra-operatively. No follow-up was presented to describe further results obtained with that device. In this scenario, it was believed that an instrument that could measure intra-operatively the stability achieved by an implanted stem would consistently improve the rate of success. This instrument should be accurate and should give to the surgeon during implantation a quick answer concerning the stability of the implanted stem. With this aim, an intra-operative device was designed, developed and validated. The device is meant to help the surgeon to decide how much to press-fit the implant. It is essentially made of a torsional load cell, able to measure the extent of torque applied by the surgeon to test primary stability, an angular sensor that measure the relative angular displacement between stem and femur, a rigid connector that enable connecting the device to the stem, and all the electronics for signals conditioning. The device was successfully validated in-vitro, showing a good overall accuracy in discriminating stable from unstable implants. Repeatability tests showed that the device was reliable. A calibration procedure was then performed in order to convert the angular readout into a linear displacement measurement, which is an information clinically relevant and simple to read in real-time by the surgeon. The second study reported in my thesis, concerns the evaluation of the possibility to have predictive information regarding the primary stability of a cementless stem, by measuring the micromotion of the last rasp used by the surgeon to prepare the femoral canal. This information would be really useful to the surgeon, who could check prior to the implantation process if the planned stem size can achieve a sufficient degree of primary stability, under optimal press fitting conditions. An intra-operative tool was developed to this aim. It was derived from a previously validated device, which was adapted for the specific purpose. The device is able to measure the relative micromotion between the femur and the rasp, when a torsional load is applied. An in-vitro protocol was developed and validated on both composite and cadaveric specimens. High correlation was observed between one of the parameters extracted form the acquisitions made on the rasp and the stability of the corresponding stem, when optimally press-fitted by the surgeon. After tuning in-vitro the protocol as in a closed loop, verification was made on two hip patients, confirming the results obtained in-vitro and highlighting the independence of the rasp indicator from the bone quality, anatomy and preserving conditions of the tested specimens, and from the sharpening of the rasp blades. The third study is related to an approach that have been recently explored in the orthopaedic community, but that was already in use in other scientific fields. It is based on the vibration analysis technique. This method has been successfully used to investigate the mechanical properties of the bone and its application to evaluate the extent of fixation of dental implants has been explored, even if its validity in this field is still under discussion. Several studies have been published recently on the stability assessment of hip implants by vibration analysis. The aim of the reported study was to develop and validate a prototype device based on the vibration analysis technique to measure intra-operatively the extent of implant stability. The expected advantages of a vibration-based device are easier clinical use, smaller dimensions and minor overall cost with respect to other devices based on direct micromotion measurement. The prototype developed consists of a piezoelectric exciter connected to the stem and an accelerometer attached to the femur. Preliminary tests were performed on four composite femurs implanted with a conventional stem. The results showed that the input signal was repeatable and the output could be recorded accurately. The fourth study concerns the application of the device based on the vibration analysis technique to several cases, considering both composite and cadaveric specimens. Different degrees of bone quality were tested, as well as different femur anatomies and several levels of press-fitting were considered. The aim of the study was to verify if it is possible to discriminate between stable and quasi-stable implants, because this is the most challenging detection for the surgeon in the operation room. Moreover, it was possible to validate the measurement protocol by comparing the results of the acquisitions made with the vibration-based tool to two reference measurements made by means of a validated technique, and a validated device. The results highlighted that the most sensitive parameter to stability is the shift in resonance frequency of the stem-bone system, showing high correlation with residual micromotion on all the tested specimens. Thus, it seems possible to discriminate between many levels of stability, from the grossly loosened implant, through the quasi-stable implants, to the definitely stable one. Finally, an additional study was performed on a different type of hip prosthesis, which has recently gained great interest thus becoming fairly popular in some countries in the last few years: the hip resurfacing prosthesis. The study was motivated by the following rationale: although bone-prosthesis micromotion is known to influence the stability of total hip replacement, its effect on the outcome of resurfacing implants has not been investigated in-vitro yet, but only clinically. Thus the work was aimed at verifying if it was possible to apply to the resurfacing prosthesis one of the intraoperative devices just validated for the measurement of the micromotion in the resurfacing implants. To do that, a preliminary study was performed in order to evaluate the extent of migration and the typical elastic movement for an epiphyseal prosthesis. An in-vitro procedure was developed to measure micromotions of resurfacing implants. This included a set of in-vitro loading scenarios that covers the range of directions covered by hip resultant forces in the most typical motor-tasks. The applicability of the protocol was assessed on two different commercial designs and on different head sizes. The repeatability and reproducibility were excellent (comparable to the best previously published protocols for standard cemented hip stems). Results showed that the procedure is accurate enough to detect micromotions of the order of few microns. The protocol proposed was thus completely validated. The results of the study demonstrated that the application of an intra-operative device to the resurfacing implants is not necessary, as the typical micromovement associated to this type of prosthesis could be considered negligible and thus not critical for the stabilization process. Concluding, four intra-operative tools have been developed and fully validated during these three years of research activity. The use in the clinical setting was tested for one of the devices, which could be used right now by the surgeon to evaluate the degree of stability achieved through the press-fitting procedure. The tool adapted to be used on the rasp was a good predictor of the stability of the stem. Thus it could be useful for the surgeon while checking if the pre-operative planning was correct. The device based on the vibration technique showed great accuracy, small dimensions, and thus has a great potential to become an instrument appreciated by the surgeon. It still need a clinical evaluation, and must be industrialized as well. The in-vitro tool worked very well, and can be applied for assessing resurfacing implants pre-clinically.
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L’elaborazione di questa tesi è stata svolta con l’ausilio di strumenti di Business Intelligence. In particolare, si è dapprima alimentato un data warehouse territoriale, in cui sono stati inseriti, dopo averli elaborati, i dati messi a disposizione dagli osservatori territoriali nazionali e dall’azienda Geofor spa. A partire da questi, sono stati prodotti degli indicatori statistici e dei report, utili per evidenziare andamenti e trend di crescita di alcuni particolari indici. Il principale strumento utilizzato è StatPortal, un portale Web di Business Intelligence OLAP per la realizzazione di Data warehouse territoriali. L’argomento sarà approfondito nel capitolo sette, dedicato agli strumenti utilizzati, ma in breve, questo sistema consente di raccogliere, catalogare e condividere informazione statistica e geostatistica, nonché di produrre indicatori e reportistica. Il lavoro è organizzato come segue: inizialmente c’è una prima parte di definizione e classificazione dei rifiuti che ha l’obiettivo di permettere al lettore di inquadrare il tema e prendere coscienza del problema. Successivamente, è stata sviluppata una parte più storica, con una rapida analisi temporale per comprendere il “tipping point”, cioè il momento in cui i rifiuti hanno iniziato a essere percepiti come un problema per la comunità, concludendo con un accenno agli scenari attuali e futuri. In seguito, si è indirizzata l’attenzione sul panorama italiano, europeo e mondiale citando alcuni interessanti e originali esempi di efficienza nella gestione dei rifiuti, che potrebbero servire da spunto per qualche stakeholder nazionale. Si è poi introdotta quella che è la normativa vigente, sottolineando quali sono gli obiettivi che impone ed entro quali tempi dovranno essere raggiunti, elencando quindi i principi fondamentali del D.lgs.152/2006 e del D.lgs 36/2003. Continuando su questo filo logico, si è voluto introdurre al lettore, la questione dei Rifiuti Solidi Urbani (RSU) nel Comune di Pisa. Sono stati definiti: lo stato dell’arte dell’igiene urbana pisana, i sistemi implementati nella città con i vari pregi e difetti e quali sono state le azioni pratiche messe in atto dall’Amministrazione per far fronte al tema. Il capitolo sei rappresenta uno dei due punti focali dell’intero lavoro: il Rapporto sullo Stato dell’Ambiente della città di Pisa in tema di rifiuti urbani. Qui saranno analizzati i vari indici e report prodotti ad hoc con lo strumento Statportal appena menzionato, con lo scopo di segnalare evidenze e obiettivi dell’Amministrazione. Nel settimo capitolo si analizza la fase di progettazione del Data Warehouse. Sono elencati i passi fondamentali nella costruzione di un DW dimensionale, esponendone in primo luogo la specifica dei requisiti del progetto ed elencando per ognuno di essi le dimensioni, le misure e le aggregazioni relative. In seguito saranno descritti nel dettaglio la fase di progettazione concettuale e lo schema logico. In ultimo, sarà presentato l’altro punto focale di questa tesi, nonché la parte più interattiva: un portale web creato appositamente per il Comune con l’obiettivo di coinvolgere ed aiutare i cittadini nel conferimento dei rifiuti da loro prodotti. Si tratta di una sorta di manuale interattivo per individuare come eseguire una corretta differenziazione dei rifiuti. Lo scopo primario è quello di fare chiarezza alle utenze nella differenziazione, il che, in maniera complementare, dovrebbe incrementare la qualità del rifiuto raccolto, minimizzando i conferimenti errati. L’obiettivo principale di questo lavoro resta quindi il monitoraggio e l’analisi delle tecniche e dei processi di gestione dei rifiuti nel Comune di Pisa. Analogamente si vuole coinvolgere e suscitare l’interesse del maggior numero di persone possibile al tema della sostenibilità ambientale, rendendo consapevole il lettore che il primo passo verso un mondo più sostenibile spetta in primis a Noi che quotidianamente acquistiamo, consumiamo ed infine gettiamo via i residui senza troppo preoccuparci. Il fatto che anche in Italia, si stia sviluppando un senso civico e una forte responsabilizzazione verso l’ambiente da parte dei cittadini, fa ben sperare. Questo perché si è riusciti a imprimere il concetto che le soluzioni si ottengano impegnandosi in prima persona. E’ alla nostra comunità che si affida il dovere di non compromettere l’esistenza delle generazioni future, incaricandola del compito di ristabilire un equilibrio, ormai precario, tra umanità e ambiente, se non altro perché, come recita un vecchio proverbio Navajo: “il mondo non lo abbiamo in eredità dai nostri padri ma lo abbiamo in prestito dai nostri figli”.
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Die vorliegende Arbeit behandelt die Adsorption von Phospholipidvesikeln, Proteinen und Latex Beads an funktionalisierten festkörpergestützten Oberflächen. Der Schwerpunkt der Abhandlung liegt in der Modellierung der Kinetik mit Hilfe eines Simulationsprogramms unter Berücksichtigung des Massentransports in Staupunktgeometrie, der Adsorption und nachfolgender konformativer Änderungen adhärierter Partikel. Das Simulationsprogramm basiert auf einem RSA (random sequential adsorption) Algorithmus und erlaubt es aufgrund zusätzlich implementierter Optionen, z.B. zur Behandlung elektrostatischer Abstoßung, von Spreitprozessen oder von Desorptionsereignissen, den Adsorptionsprozess unter realistischen physikalischen Bedingungen nachzuempfinden und spezifische biologische Fragestellungen zu beantworten. Aus Anpassungen von Simulationen an experimentelle Daten ergeben sich dynamische Parameter, wie z.B. die Transport-, Adsorptions-, Spreit- und die Desorptionsrate. Die experimentellen Daten wurden mit Hilfe der Quarzmikrowaagetechnik (QCM), der Impedanzspektroskopie (IS) und der Rasterkraftmikroskopie (AFM) erhoben. Zusätzlich zur Kinetik gibt die graphische Ausgabe des Simulationsprogramms Aufschluss über die Oberflächenverteilung der adsorbierten und gespreiteten Partikel. Bei der Untersuchung von Systemen, in denen die Adsorption reversibel ist und infolge dessen Desorptionsprozesse eine wichtige Rolle spielen, wurde mit Hilfe des Simulationsprogramms ein völlig neuartiges Sensorkonzept in Aussicht gestellt. Es wurde gezeigt, dass aus der Analyse des Leistungsspektrums der Fluktuationen in der Belegung die Kinetik von Adsorption und Desorption ermittelt werden kann. Dies ist aus sensorischer Sicht interessant, da eine Vielzahl störender Einflüsse, wie z.B: der Massentransport und die elektronische Drift in diesem Fall nicht berücksichtigt werden müssen.
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Im ersten Teil der Arbeit wurde das Bindungsverhalten von Annexin A1 und Annexin A2t an festkörperunterstützte Lipidmembranen aus POPC und POPS untersucht. Für beide Proteine konnte mit Hilfe der Fluoreszenzmikroskopie gezeigt werden, dass irreversible Bindung nur in Anwesenheit von POPS auftritt. Durch rasterkraftmikroskopische Aufnahmen konnte die laterale Organisation der Annexine auf der Lipidmembran dargestellt werden. Beide Proteine lagern sich in Form lateraler Aggregate (zweidimensionale Domänen) auf der Oberfläche an, außerdem ist der Belegungsgrad und die Größe der Domänen von der Membranzusammensetzung und der Calciumkonzentration abhängig. Mit zunehmendem POPS-Gehalt und Calciumkonzentration steigt der Belegungsgrad an und der mittlere Domänenradius wird kleiner. Diese Ergebnisse konnten in Verbindung mit detaillierten Bindungsstudien des Annexins A1 mit der Quarzmikrowaage verwendet werden, um ein Bindungsmodell auf Basis einer heterogenen Oberfläche zu entwickeln. Auf einer POPC-reichen Matrix findet reversible Adsorption statt und auf POPS-reichen Domänen irreversible Adsorption. Durch die Anpassung von dynamischen Monte Carlo-Simulationen basierend auf einer zweidimensionalen zufälligen sequentiellen Adsorption konnten Erkenntnisse über die Membranstruktur und die kinetischen Ratenkonstanten in Abhängigkeit von der Calciumkonzentration und der Inkubationszeit des Proteins gewonnen werden. Die irreversible Bindung ist in allen Calciumkonzentrationsbereichen schneller als die reversible. Außerdem zeigt die irreversible Adsorption eine deutlich stärkere Abhängigkeit von der Calciumkonzentration. Ein kleinerer Belegungsgrad bei niedrigen Ca2+-Gehalten ist hauptsächlich durch die Abnahme der verfügbaren Bindungsplätze auf der Oberfläche zu erklären. Die gute Übereinstimmung der aus den Monte Carlo-Simulationen erhaltenen Domänenstrukturen mit den rasterkraftmikroskopischen Aufnahmen und die Tatsache, dass sich die simulierten Resonanzfrequenzverläufe problemlos an die experimentellen Kurven aus den QCM-Messungen anpassen ließen, zeigt die gute Anwendbarkeit des entwickelten Simulationsprogramms auf die Adsorption von Annexin A1. Die Extraktion der kinetischen Parameter aus dem zweidimensionalen RSA-Modell ist mit Sicherheit einem einfachen Langmuir-Ansatz überlegen. Bei einem Langmuir-Modell erfolgt eine integrale Erfassung einer einzelnen makroskopischen Geschwindigkeitskonstante, während durch das RSA-Modell eine differenzierte Betrachtung des reversiblen und irreversiblen Bindungsprozesses möglich ist. Zusätzlich lassen sich mikroskopische Informationen über die Oberflächenbeschaffenheit gewinnen. Im zweiten Teil der Arbeit wurde das thermotrope Phasenverhalten von festkörperunterstützten Phospholipidbilayern untersucht. Dazu wurden mikrostrukturierte, frei stehende Membranstreifen präpariert und mit Hilfe der bildgebenden Ellipsometrie untersucht. Dadurch konnten die temperaturabhängigen Verläufe der Schichtdicke und der lateralen Membranausdehnung parallel beobachtet werden. Die ermittelten Phasenübergangstemperaturen von DMPC, diC15PC und DPPC lagen 2 - 3 °C oberhalb der Literaturwerte für vesikuläre Systeme. Außerdem wurde eine deutliche Verringerung der Kooperativität der Phasenumwandlung gefunden, was auf einen großen Einfluss des Substrats bei den festkörperunterstützten Lipidmembranen schließen lässt. Zusätzlich wurde ein nicht systematischer Zusammenhang der Ergebnisse von der Oberflächenpräparation gefunden, der es unabdingbar macht, bei Untersuchungen von festkörperunterstützten Substraten einen internen Standard einzuführen. Bei der Analyse des thermotropen Phasenübergangsverhaltens von DMPC/Cholesterol - Gemischen wurde daher die individuelle Adressierbarkeit der strukturierten Lipidmembranen ausgenutzt und ein Lipidstreifen aus reinem DMPC als Standard verwendet. Auf diese Weise konnte gezeigt werden, dass das für Phospholipide typische Phasenübergangsverhalten ab 30 mol% Cholesterol in der Membran nicht mehr vorhanden ist. Dies ist auf die Bildung einer nur durch höhere Sterole induzierten fluiden Phase mit hoch geordneten Acylketten zurückzuführen. Abschließend konnte durch die Zugabe von Ethanol zu einer mikrostrukturierten DMPC-Membran die Bildung eines interdigitierten Bilayers nachgewiesen werden. Die bildgebende Ellipsometrie ist eine sehr gute Methode zur Untersuchung festkörperunterstützter Lipidmembranen, da sie über ein sehr gutes vertikales und ein ausreichendes laterales Auflösungsvermögen besitzt. Sie ist darin zwar einem Rasterkraftmikroskop noch unterlegen, besitzt dafür aber eine einfachere Handhabung beim Umgang mit Flüssigkeiten und in der Temperierung, eine schnellere Bildgebung und ist als optische Methode nicht-invasiv.
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Le reti di oggetti intelligenti costituiscono una realtà che si sta affermando nel mondo quotidiano. Dispositivi capaci di comunicare tra loro, oltre che svolgere la propria funzione primaria, possono comporre una nuvola che faccia riferimento al legittimo proprietario. Un aspetto fondamentale di questo scenario riguarda la sicurezza, in particolar modo per garantire una comunicazione protetta. Il soddisfacimento di questo requisito è fondamentale anche per altri punti come l'integrità dell'informazione condivisa e l'autenticazione. Lo strumento più antico e tutt'ora adatto alla riservatezza di una comunicazione è costituito dalla crittografia. Una tecnica crittografica è schematicamente composta da un algoritmo che, a seconda di una chiave e del messaggio in ingresso, restituisce in uscita un messaggio cifrato, il crittogramma. Questo viene poi inviato e al legittimo destinatario che, essendo in possesso della chiave e dell'algoritmo, lo converte nel messaggio originale. L'obiettivo è rendere impossibile ad un utente malevolo - non dotato di chiave - la ricostruzione del messaggio. L'assunzione che l'algoritmo possa essere noto anche a terze parti concentra l'attenzione sul tema della chiave. La chiave deve essere sufficientemente lunga e casuale, ma soprattutto deve essere nota ai due utenti che intendono instaurare la comunicazione. Quest'ultimo problema, noto come distribuzione della chiave, è stato risolto con il sistema RSA a chiave pubblica. Il costo computazionale di questa tecnica, specialmente in un contesto di dispositivi non caratterizzati da grandi potenze di calcolo, suggerisce però la ricerca di strade alternative e meno onerose. Una soluzione promettente ed attualmente oggetto di studio sembra essere costituita dalle proprietà del canale wireless. Un ponte radio è caratterizzato da una funzione di trasferimento che dipende dall'ambiente in cui ci si trova e, per il teorema di reciprocità, risulta essere lo stesso per i due utenti che l'hanno instaurato. Oggetto della tesi è lo studio ed il confronto di alcune delle tecniche possibili per estrarre una chiave segreta da un mezzo condiviso, come quello del canale wireless. Si presenterà il contesto in cui verrà sviluppato l'elaborato. Si affronteranno in particolare due casi di interesse, costituiti dalla attuale tecnologia di propagazione del segnale a banda stretta (impiegata per la maggior parte delle trasmissioni senza fili) per passare a quella relativamente più recente della banda Ultra-larga (UWB). Verranno poi illustrate delle tecniche per ottenere stringhe di bit dai segnali acquisiti, e saranno proposti dei metodi per la loro correzione da eventuali discordanze. Saranno infine riportate le conclusioni sul lavoro svolto e le possibili strade future.
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Le tecniche di next generation sequencing costituiscono un potente strumento per diverse applicazioni, soprattutto da quando i loro costi sono iniziati a calare e la qualità dei loro dati a migliorare. Una delle applicazioni del sequencing è certamente la metagenomica, ovvero l'analisi di microorganismi entro un dato ambiente, come per esempio quello dell'intestino. In quest'ambito il sequencing ha permesso di campionare specie batteriche a cui non si riusciva ad accedere con le tradizionali tecniche di coltura. Lo studio delle popolazioni batteriche intestinali è molto importante in quanto queste risultano alterate come effetto ma anche causa di numerose malattie, come quelle metaboliche (obesità, diabete di tipo 2, etc.). In questo lavoro siamo partiti da dati di next generation sequencing del microbiota intestinale di 5 animali (16S rRNA sequencing) [Jeraldo et al.]. Abbiamo applicato algoritmi ottimizzati (UCLUST) per clusterizzare le sequenze generate in OTU (Operational Taxonomic Units), che corrispondono a cluster di specie batteriche ad un determinato livello tassonomico. Abbiamo poi applicato la teoria ecologica a master equation sviluppata da [Volkov et al.] per descrivere la distribuzione dell'abbondanza relativa delle specie (RSA) per i nostri campioni. La RSA è uno strumento ormai validato per lo studio della biodiversità dei sistemi ecologici e mostra una transizione da un andamento a logserie ad uno a lognormale passando da piccole comunità locali isolate a più grandi metacomunità costituite da più comunità locali che possono in qualche modo interagire. Abbiamo mostrato come le OTU di popolazioni batteriche intestinali costituiscono un sistema ecologico che segue queste stesse regole se ottenuto usando diverse soglie di similarità nella procedura di clustering. Ci aspettiamo quindi che questo risultato possa essere sfruttato per la comprensione della dinamica delle popolazioni batteriche e quindi di come queste variano in presenza di particolari malattie.
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Root-yield-1.06 is a major QTL affecting root system architecture (RSA) and other agronomic traits in maize. The effect of this QTL has been evaluated with the development of near isogenic lines (NILs) differing at the QTL position. The objective of this study was to fine map qroot-yield-1.06 by marker-assisted searching for chromosome recombinants in the QTL interval and concurrent root phenotyping in both controlled and field conditions, through successive generations. Complementary approaches such as QTL meta-analysis and RNA-seq were deployed in order to help prioritizing candidate genes within the QTL target region. Using a selected group of genotypes, field based root analysis by ‘shovelomics’ enabled to accurately collect RSA information of adult maize plants. Shovelomics combined with software-assisted root imaging analysis proved to be an informative and relatively highly automated phenotyping protocol. A QTL interval mapping was conducted using a segregating population at the seedling stage grown in controlled environment. Results enabled to narrow down the QTL interval and to identify new polymorphic markers for MAS in field experiments. A collection of homozygous recombinant NILs was developed by screening segregating populations with markers flanking qroot-yield-1.06. A first set of lines from this collection was phenotyped based on the adapted shovelomics protocol. QTL analysis based on these data highlighted an interval of 1.3 Mb as completely linked with the target QTL but, a larger safer interval of 4.1 Mb was selected for further investigations. QTL meta-analysis allows to synthetize information on root QTLs and two mQTLs were identified in the qroot-yield-1.06 interval. Trascriptomics analysis based on RNA-seq data of the two contrasting QTL-NILs, confirmed alternative haplotypes at chromosome bin 1.06. qroot-yield-1.06 has now been delimited to a 4.1-Mb interval, and thanks to the availability of additional untested homozygous recombinant NILs, the potentially achievable mapping resolution at qroot-yield-1.06 is c. 50 kb.