RNA Interference and cyclooxygenase-2 (COX-2) regulation in colon cancer cells

Despite new methods and combined strategies, conventional cancer chemotherapy still lacks specificity and induces drug resistance. Gene therapy can offer the potential to obtain the success in the clinical treatment of cancer and this can be achieved by replacing mutated tumour suppressor genes, inhibiting gene transcription, introducing new genes encoding for therapeutic products, or specifically silencing any given target gene. Concerning gene silencing, attention has recently shifted onto the RNA interference (RNAi) phenomenon. Gene silencing mediated by RNAi machinery is based on short RNA molecules, small interfering RNAs (siRNAs) and microRNAs (miRNAs), that are fully o partially homologous to the mRNA of the genes being silenced, respectively. On one hand, synthetic siRNAs appear as an important research tool to understand the function of a gene and the prospect of using siRNAs as potent and specific inhibitors of any target gene provides a new therapeutical approach for many untreatable diseases, particularly cancer. On the other hand, the discovery of the gene regulatory pathways mediated by miRNAs, offered to the research community new important perspectives for the comprehension of the physiological and, above all, the pathological mechanisms underlying the gene regulation. Indeed, changes in miRNAs expression have been identified in several types of neoplasia and it has also been proposed that the overexpression of genes in cancer cells may be due to the disruption of a control network in which relevant miRNA are implicated. For these reasons, I focused my research on a possible link between RNAi and the enzyme cyclooxygenase-2 (COX-2) in the field of colorectal cancer (CRC), since it has been established that the transition adenoma-adenocarcinoma and the progression of CRC depend on aberrant constitutive expression of COX-2 gene. In fact, overexpressed COX-2 is involved in the block of apoptosis, the stimulation of tumor-angiogenesis and promotes cell invasion, tumour growth and metastatization. On the basis of data reported in the literature, the first aim of my research was to develop an innovative and effective tool, based on the RNAi mechanism, able to silence strongly and specifically COX-2 expression in human colorectal cancer cell lines. In this study, I firstly show that an siRNA sequence directed against COX-2 mRNA (siCOX-2), potently downregulated COX-2 gene expression in human umbilical vein endothelial cells (HUVEC) and inhibited PMA-induced angiogenesis in vitro in a specific, non-toxic manner. Moreover, I found that the insertion of a specific cassette carrying anti-COX-2 shRNA sequence (shCOX-2, the precursor of siCOX-2 previously tested) into a viral vector (pSUPER.retro) greatly increased silencing potency in a colon cancer cell line (HT-29) without activating any interferon response. Phenotypically, COX-2 deficient HT-29 cells showed a significant impairment of their in vitro malignant behaviour. Thus, results reported here indicate an easy-to-use, powerful and high selective virus-based method to knockdown COX-2 gene in a stable and long-lasting manner, in colon cancer cells. Furthermore, they open up the possibility of an in vivo application of this anti-COX-2 retroviral vector, as therapeutic agent for human cancers overexpressing COX-2. In order to improve the tumour selectivity, pSUPER.retro vector was modified for the shCOX-2 expression cassette. The aim was to obtain a strong, specific transcription of shCOX-2 followed by COX-2 silencing mediated by siCOX-2 only in cancer cells. For this reason, H1 promoter in basic pSUPER.retro vector [pS(H1)] was substituted with the human Cox-2 promoter [pS(COX2)] and with a promoter containing repeated copies of the TCF binding element (TBE) [pS(TBE)]. These promoters were choosen because they are partculary activated in colon cancer cells. COX-2 was effectively silenced in HT-29 and HCA-7 colon cancer cells by using enhanced pS(COX2) and pS(TBE) vectors. In particular, an higher siCOX-2 production followed by a stronger inhibition of Cox-2 gene were achieved by using pS(TBE) vector, that represents not only the most effective, but also the most specific system to downregulate COX-2 in colon cancer cells. Because of the many limits that a retroviral therapy could have in a possible in vivo treatment of CRC, the next goal was to render the enhanced RNAi-mediate COX-2 silencing more suitable for this kind of application. Xiang and et al. (2006) demonstrated that it is possible to induce RNAi in mammalian cells after infection with engineered E. Coli strains expressing Inv and HlyA genes, which encode for two bacterial factors needed for successful transfer of shRNA in mammalian cells. This system, called “trans-kingdom” RNAi (tkRNAi) could represent an optimal approach for the treatment of colorectal cancer, since E. Coli in normally resident in human intestinal flora and could easily vehicled to the tumor tissue. For this reason, I tested the improved COX-2 silencing mediated by pS(COX2) and pS(TBE) vectors by using tkRNAi system. Results obtained in HT-29 and HCA-7 cell lines were in high agreement with data previously collected after the transfection of pS(COX2) and pS(TBE) vectors in the same cell lines. These findings suggest that tkRNAi system for COX-2 silencing, in particular mediated by pS(TBE) vector, could represent a promising tool for the treatment of colorectal cancer. Flanking the studies addressed to the setting-up of a RNAi-mediated therapeutical strategy, I proposed to get ahead with the comprehension of new molecular basis of human colorectal cancer. In particular, it is known that components of the miRNA/RNAi pathway may be altered during the progressive development of colorectal cancer (CRC), and it has been already demonstrated that some miRNAs work as tumor suppressors or oncomiRs in colon cancer. Thus, my hypothesis was that overexpressed COX-2 protein in colon cancer could be the result of decreased levels of one or more tumor suppressor miRNAs. In this thesis, I clearly show an inverse correlation between COX-2 expression and the human miR- 101(1) levels in colon cancer cell lines, tissues and metastases. I also demonstrate that the in vitro modulating of miR-101(1) expression in colon cancer cell lines leads to significant variations in COX-2 expression, and this phenomenon is based on a direct interaction between miR-101(1) and COX-2 mRNA. Moreover, I started to investigate miR-101(1) regulation in the hypoxic environment since adaptation to hypoxia is critical for tumor cell growth and survival and it is known that COX-2 can be induced directly by hypoxia-inducible factor 1 (HIF-1). Surprisingly, I observed that COX-2 overexpression induced by hypoxia is always coupled to a significant decrease of miR-101(1) levels in colon cancer cell lines, suggesting that miR-101(1) regulation could be involved in the adaption of cancer cells to the hypoxic environment that strongly characterize CRC tissues.

L'introduzione dell'e-commerce nel settore fieristico: piano di email marketing e studio di usabilità

L’obiettivo di questa relazione è presentare i risultati raggiunti dalle attività di marketing svolte al fine di promuovere e valutare la piattaforma e-commerce di BF Servizi, azienda che opera come intermediario nella fornitura di servizi all’interno del contesto fieristico di Bologna Fiere. A fine del 2009 BF Servizi ha commissionato a Gecod lo sviluppo di un sistema informativo per le gestione integrata di tutti i processi rilevanti per il business aziendale. A ottobre 2010 viene completato e presentato uno dei moduli previsti dall’intero progetto: la piattaforma e-commerce. Questa si differenzia dagli altri sistemi di commercio online in quanto è stata progettata specificatamente allo scopo di permettere la prenotazione di servizi da parte delle aziende che partecipano alle manifestazioni svolte presso i padiglioni di Bologna Fiere. Le sue particolarità sono quindi sia l’utenza, che configura la creazione di relazioni B2B, sia la tipologia di articoli offerti, non prodotti ma servizi, sia l’ambiente, le fiere svolte a Bologna. A seguito di questo rinnovamento BF Servizi ha ritenuto opportuno adeguare al nuovo contesto anche le proprie tecniche di marketing introducendo, come strumento da usare per la loro attuazione, il web. All’interno dell’organico dell’azienda però non era presente una figura che avesse le competenze per poter integrare, ai normali canali usati per le iniziative di marketing, le opportunità offerte da Internet e che quindi potesse utilizzarle per raggiungere gli obiettivi prefissati per tali attività e cioè la promozione della piattaforma e-commerce e la misurazione del suo gradimento da parte dell’utenza. Le mie conoscenze maturate in ambito universitario hanno apportato quel contributo necessario alla loro realizzazione. La prima decisione è stata la scelta di quale tra le tecniche di web marketing fosse la migliore per le promozione della piattaforma e-commerce di BF Servizi. La disponibilità di un voluminoso numero di indirizzi email dei loro clienti e prospect ed una generale bassa alfabetizzazione informatica degli stessi ha portato a individuare quale soluzione più adeguata la tecnica dell’email marketing. La comunicazione ai clienti per mezzo email era già prevista ma senza che vi fosse ne una pianificazione ne un'analisi dei risultati. Era quindi necessario formalizzare un vero e proprio piano di marketing e successivamente migliorarne le performance. L’assenza di strumenti adeguati alla sua realizzazione ha richiesto che prima venisse svolto un benchmark del settore basato sul confronto di funzionalità, costi e facilità d'uso. Il servizio ESP di Mailchimp è risultato il vincitore in quanto assicurava una maggiore tutela della privacy, una migliore gestione dello spam e strumenti più efficaci per la creazione e l'analisi delle campagne email. Per la valutazione del gradimento della piattaforma e-commerce da parte degli utenti è stato deciso di progettare un’apposita indagine sottoforma di Usability Test e Online Survey. Per il primo è stato necessario l’utilizzo di un apposito servizio per la condivisione dello schermo in quanto in era impossibile far ritrovare il test e l’osservatore nel medesimo luogo. Per l’Online Survey, grazie ad un piccolo Benchmark, è stato deciso di utilizzare SurveyGizmo. All’interno dell’organico dell’azienda non è presente una figura di importanza strategia quale il Web Marketing Manager che abbia la responsabilità di coordinare tutte le attività legate alla vendita o al lancio di un prodotto o di un servizio. L’attività di marketing viene svolta da una figura interna all’azienda (il Responsabile Marketing) che si avvale della collaborazione sia di Gecod, per quanto riguarda l'area informatica, sia di BitBang per quanto riguarda alcune attività di web marketing. La realizzazione delle due attività ha fortemente risentito di questa frammentazione dei compiti, è stato infatti opportuno adeguare le pianificazione alle esigenze di Gecod. Nonostante questo i dati conseguiti dall'attività di email marketing sono stati utili a capire l'efficacia e l'efficienza di questa tecnica nel contesto di BF Servizi. Anche le informazioni raccolte durante l'indagine sono state sufficienti per poter dare un valore all'usabilità della piattaforma e-commerce. La pianificazione iniziale di un piano di email marketing richiede che vengano svolte varie verifiche per valutare la qualità ed inoltre richiede periodo medio-lungo per poter essere reso ottimale. Era quindi già chiaro sin dall'inizio come uno degli obiettivi fosse quello di progettare una buona base su cui apportare successive modifiche. Le prime campagne realizzate sarebbero state utili a ricavare le informazioni necessarie per correggere il piano di marketing. La tipologia di relazione che si instaura tra BF Servizi e i suoi clienti è particolare, la gran parte di essi infatti non vi mantiene un rapporto continuativo durante l’anno ma lo esaurisce in un unico momento poco prima del manifestarsi dell’evento fieristico. L'attenzione dei clienti verso le comunicazioni di BF Servizi è elevata solo in questo periodo e la pianificazione delle campagne ha dovuto tener conto di questo aspetto. Sono state realizzate le campagne solo per gli espositori che partecipavano agli eventi fieristici di Marca 2011, Artefiera 2011 e Fishing Show 2011, cioè le manifestazioni che si sono svolte durante il mio periodo di lavoro in azienda. Le performance delle campagne sono state buone e i dati raccolti hanno permesso di identificare quali siano gli aspetti da migliorare per poter renderle ancora migliori.

Analisi, progettazione e valutazione dell'efficacia dello spot pubblicitario. Una semiotica per la ricerca di mercato

Il lavoro di ricerca tenta di inquadrare sotto nuove prospettive una problematica ormai classica all’interno della semiotica della pubblicità: l’analisi dello spot. I punti chiave del lavoro – e la pretesa di una certa differenza rispetto a lavori con oggetti affini – consistono sostanzialmente in tre aspetti. Innanzitutto, vi è un ritorno alle origini flochiane nella misura in cui non solo il contesto complessivo e le finalità che la ricerca si propone sono fortemente ancorati all’interno di obiettivi di marketing, ma tutto lo studio nella sua interezza nasce dal dialogo concreto tra metodologia di analisi semiotica e prassi concreta all’interno degli istituti di ricerca di mercato. La tesi non presenta quindi una collezione di analisi di testi pubblicitari condotte in modo autoriferito, quanto piuttosto di “messe alla prova” della metodologia, funzionali alla definizione di disegni di ricerca per la marketing research. Questo comporta un dialogo piuttosto stretto con metodologie affini (sociologia qualitativa e quantitativa, psicologia motivazionale, ecc.) nella convinzione che la priorità accordata all’oggetto di analisi sia sovraordinata rispetto all’ortodossia degli strumenti metodologici. In definitiva, lo spot è sempre e comunque analizzato all’interno di una prospettiva brand-centrica che ha ben in mente la semiotica della situazione di consumo rispetto alla quale lo spot agisce da leva di valorizzazione per l’acquisto. In secondo luogo, gli oggetti analizzati sono piuttosto vari e differenziati: non solo lo spot nella sua versione audiovisiva definitiva (il “girato”), ma anche storyboard, animatic, concept (di prodotto e di comunicazione). La prospettiva generativa greimasiana va a innestarsi su problematiche legate (anche) alla genesi dello spot, alla sua progettazione e riprogettazione/ottimizzazione. La tesi mostra quindi come una semiotica per le consulenze di marketing si diriga sul proprio oggetto ponendogli domande ben circoscritte e finalizzate a un obiettivo specifico, sostanzialmente derivato dal brief contenente le intenzioni comunicazionali del cliente-azienda. Infine, pur rimanendo all’interno di una teoria semiotica generativa (sostanzialmente greimasiana e post greimasiana), la ricerca adotta una prospettiva intrinsecamente multidisciplinare che se da un lato guarda a problematiche legate al marketing, al branding e alla comunicazione pubblicitaria e d’impresa tout court, dall’altro ritorna alle teorie dell’audiovisivo, mostrando affinità e differenze rispetto a forme audiovisive standard (il “film”) e a mutuazioni da nuove estetiche (la neotelevisione, il videoclip, ecc). La tesi si mostra solidamente convinta del fatto che per parlare di efficacia discorsiva sia imprescindibile approfondire le tematiche riguardanti il sincretismo espressivo e le specifiche modalità di manifestazione stilistica. In questo contesto, il lavoro si compone di quattro grandi aree tematiche. Dopo una breve introduzione sull’attualità del tema “spot” e sulla prospettiva analiticometodologica adottata (§ 1.), nel secondo capitolo si assume teoreticamente che i contenuti dello spot derivino da una specifica (e di volta in volta diversa) creolizzazione tra domini tematici derivanti dalla marca, dal prodotto (inteso tanto come concept di prodotto, quanto come prodotto già “vestito” di una confezione) e dalle tendenze socioculturali. Le tre dimensioni vengono valutate in relazione all’opposizione tra heritage, cioè continuità rispetto al passato e ai concorrenti e vision, cioè discontinuità rispetto alla propria storia comunicazionale e a quella dei concorrenti. Si esplorano inoltre altri fattori come il testimonial-endorser che, in quanto elemento già intrinsecamente foriero di elementi di valorizzazione, va a influire in modo rilevante sul complesso tematico e assiologico della pubblicità. Essendo la sezione della tesi che prende in considerazione il piano specificatamente contenutistico dello spot, questa parte diventa quindi anche l’occasione per ritornare sul modello delle assiologie del consumo di Jean-Marie Floch, approntando alcune critiche e difendendo invece un modello che – secondo la prospettiva qui esposta – contiene punti di attualità ineludibili rispetto a schematizzazioni che gli sono successive e in qualche modo debitrici. Segue una sezione (§ 3.) specificatamente dedicata allo svolgimento e dis-implicazione del sincretismo audiovisivo e quindi – specularmente alla precedente, dedicata alle forme e sostanze del contenuto – si concentra sulle dinamiche espressive. Lo spot viene quindi analizzato in quanto “forma testuale” dotata di alcune specificità, tra cui in primis la brevità. Inoltre vengono approfondite le problematiche legate all’apporto di ciascuna specifica sostanza: il rapporto tra visivo e sonoro, lo schermo e la sua multiprospetticità sempre più evidente, il “lavoro” di punteggiatura della musica, ecc. E su tutto il concetto dominante di montaggio, intrinsecamente unito a quello di ritmo. Il quarto capitolo ritorna in modo approfondito sul rapporto tra semiotica e ricerca di mercato, analizzando sia i rapporti di reciproca conoscenza (o non conoscenza), sia i nuovi spazi di intervento dell’analisi semiotica. Dopo aver argomentato contro un certo scetticismo circa l’utilità pragmatica dell’analisi semiotica, lo studio prende in esame i tradizionali modelli di valutazione e misurazione dell’efficacia pubblicitaria (pre- e post- test) cercando di semiotizzarne il portato. Ne consegue la proposta di disegni di ricerca semiotici modulari: integrabili tra loro e configurabili all’interno di progetti semio-quali-quantitativi. Dopo aver ridefinito le possibilità di un’indagine semiotica sui parametri di efficacia discorsiva, si procede con l’analisi di un caso concreto (§ 5.): dato uno spot che si è dimostrato efficace agli occhi dell’azienda committente, quali possono essere i modi per replicarne i fattori di successo? E come spiegare invece quelli di insuccesso delle campagne successive che – almeno teoricamente – erano pensate per capitalizzare l’efficacia della prima? Non si tratta quindi di una semiotica ingenuamente chiamata a “misurare” l’efficacia pubblicitaria, che evidentemente la marketing research analizza con strumenti quantitativi assodati e fondati su paradigmi di registrazione di determinati parametri sul consumatore (ricordo spontaneo e sollecitato, immagine di marca risultante nella mente di user e prospect consumer, intenzione d’acquisto stimolata). Piuttosto l’intervento qui esposto si preoccupa più funzionalmente a spiegare quali elementi espressivi, discorsivi, narrativi, siano stati responsabili (e quindi prospetticamente potranno condizionare in positivo o in negativo in futuro) la ricezione dello spot. L’analisi evidenzia come elementi apparentemente minimali, ancorati a differenti livelli di pertinenza siano in grado di determinare una notevole diversità negli effetti di senso. Si tratta quindi di un problema di mancata coerenza tra intenzioni comunicative e testo pubblicitario effettivamente realizzato. La risoluzione di tali questioni pragmatiche conduce ad approfondimenti teoricometodologici su alcuni versanti particolarmente interessanti. In primo luogo, ci si interroga sull’apporto della dimensione passionale nella costruzione dell’efficacia e nel coinvolgimento dello spettatore/consumatore. Inoltre – e qui risiede uno dei punti di maggior sintesi del lavoro di tesi – si intraprende una proficua discussione dei modelli di tipizzazione dei generi pubblicitari, intesi come forme discorsive. Si fanno quindi dialogare modelli diversi ma in qualche misura coestensivi e sovrapponibili come quelli di Jean Marie Floch, Guido Ferraro, Cosetta Saba e Chiara Giaccardi. Si perviene così alla costruzione di un nuovo modello sintetico, idealmente onnipervasivo e trasversale alle prospettive analizzate.

Continuity and Change in New Products

This doctoral dissertation is triggered by an emergent problem: how can firms reinvent themselves? Continuity- and change-oriented decisions fundamentally shape overtime the activities and potential revenues of organizations and other adaptive systems, but both types of actions draw upon limited resources and rely on different organizational routines and capabilities. Most organizations appear to have difficulties in making tradeoffs, so that it is easier to overinvest in one of them than to successfully achieve a mixture of both. Nevertheless, theory and empirical evidence suggest that too little of either may reduce performance, indicating a need to learn more about how organizations reconcile these tensions. In the first paper, I moved from the consideration that rapid changes in competitive environments increasingly require firms to be “ambidextrous” implementing organizational mechanisms and structures that allow continuity- and change-oriented activities to be engaged at the same time. More specifically, I show that continuity- and change-related decisions can’t be confined either inside or outside the firm, but span overtime across distinct decision domains located within and beyond the organizational boundaries. Reconciling static and dynamic perspectives of ambidexterity, I conceptualize a firm’s strategy as a bundle of decisions about product attributes and components of the production team, proposing a multidimensional and dynamic model of structural ambidexterity that explains why and how firms could manage conflicting pressures for continuity and change in the context of new products. In the second study I note how rigorous systematic evidence documenting the success of ambidextrous organizations is lacking, and there has been very little investigation of how firms deal with continuity and change in new products. How to manage the transition form a successful product to another? What to change and what to keep? Incumbents that deal with series of products over time need to update their offerings in order to have the most relevant attributes to prospect clients without disappoint the current customer base. They need to both match and anticipate consumers’ preferences, blending something old with something new to satisfy the current demand and enlarge the herd by appealing to newer audiences. This paper contributes to strategic renewal and ambidexterity-related research with the first empirically assessment of a positive consumer response to ambidexterity in new products. Also, this study provides a practical method to monitor overtime the degree to which a brand or a firm is continuity- or change- oriented and evaluate different strategy profiles across two decision domains that play a pivotal role in new products: product attributes and components of the production team.

Jasmonates and abscisic acid influence fruit ripening and plant water use: practical, physiological and morphological aspects

The aim of the present thesis was to better understand the physiological role of the phytohormones jasmonates (JAs) and abscisic acid (ABA) during fruit ripening in prospect of a possible field application of JAs and ABA to improve fruit yield and quality. In particular, the effects of exogenous application of these substances at different fruit developmental stages and under different experimental conditions were evaluated. Some aspects of the water relations upon ABA treatment were also analysed. Three fruit species, peach (Prunus persica L. Batsch), golden (Actinidia chinensis) and green kiwifruit (Actinidia deliciosa), and several of their cvs, were used for the trials. Different experimental models were adopted: fruits in planta, detached fruit, detached branches with fruit, girdled branches and micropropagated plants. The work was structured into four sets of experiments as follows: (i) Pre-harvest methyl jasmonate (MJ) application was performed at S3/S4 transition under field conditions in Redhaven peach; ethylene production, ripening index, fruit quality and shelf-life were assessed showing that MJ-treated fruit were firmer and thus less ripe than controls as confirmed by the Index of Absorbance Difference (IAD), but exhibited a shorter shelf-life due to an increase in ethylene production. Moreover, the time course of the expression of ethylene-, auxin- and other ripening-related genes was determined. Ripening-related ACO1 and ACS1 transcript accumulation was inhibited though transiently by MJ, and gene expression of the ethylene receptor ETR2 and of the ethylene-related transcription factor ERF2 was also altered. The time course of the expression of several auxin-related genes was strongly affected by MJ suggesting an increase in auxin biosynthesis, altered auxin conjugation and release as well as perception and transport; the need for a correct ethylene/auxin balance during ripening was confirmed. (ii) Pre- and post-harvest ABA applications were carried out under field conditions in Flaminia and O’Henry peach and Stark Red Gold nectarine fruit; ethylene production, ripening index, fruit quality and shelf-life were assessed. Results show that pre-harvest ABA applications increase fruit size and skin color intensity. Also post-harvest ABA treatments alter ripening-related parameters; in particular, while ethylene production is impaired in ABA-treated fruit soluble solids concentration (SSC) is enhanced. Following field ABA applications stem water potential was modified since ABA-treated peach trees retain more water. (iii) Pre- and post-harvest ABA and PDJ treatments were carried out in both kiwifruit species under field conditions at different fruit developmental stages and in post-harvest. Ripening index, fruit quality, plant transpiration, photosynthesis and stomatal conductance were assessed. Pre-harvest treatments enhance SSC in the two cvs and flesh color development in golden kiwifruit. Post-harvest applications of either ABA or ABA plus PDJ lead to increased SSC. In addition, ABA reduces gas exchanges in A. deliciosa. (iv) Spray, drench and dipping ABA treatments were performed in micropropagated peach plants and in peach and nectarine detached branches; plant water use and transpiration, biomass production and fruit dehydration were determined. In both plants and branches ABA significantly reduces water use and fruit dehydration. No negative effects on biomass production were detected. The present information, mainly arising from plant growth regulator application in a field environment, where plants have to cope with multiple biotic and abiotic stresses, may implement the perspectives for the use of these substances in the control of fruit ripening.

Finite-element discretization of 3D energy-transport equations for semiconductors

In this thesis a mathematical model was derived that describes the charge and energy transport in semiconductor devices like transistors. Moreover, numerical simulations of these physical processes are performed. In order to accomplish this, methods of theoretical physics, functional analysis, numerical mathematics and computer programming are applied. After an introduction to the status quo of semiconductor device simulation methods and a brief review of historical facts up to now, the attention is shifted to the construction of a model, which serves as the basis of the subsequent derivations in the thesis. Thereby the starting point is an important equation of the theory of dilute gases. From this equation the model equations are derived and specified by means of a series expansion method. This is done in a multi-stage derivation process, which is mainly taken from a scientific paper and which does not constitute the focus of this thesis. In the following phase we specify the mathematical setting and make precise the model assumptions. Thereby we make use of methods of functional analysis. Since the equations we deal with are coupled, we are concerned with a nonstandard problem. In contrary, the theory of scalar elliptic equations is established meanwhile. Subsequently, we are preoccupied with the numerical discretization of the equations. A special finite-element method is used for the discretization. This special approach has to be done in order to make the numerical results appropriate for practical application. By a series of transformations from the discrete model we derive a system of algebraic equations that are eligible for numerical evaluation. Using self-made computer programs we solve the equations to get approximate solutions. These programs are based on new and specialized iteration procedures that are developed and thoroughly tested within the frame of this research work. Due to their importance and their novel status, they are explained and demonstrated in detail. We compare these new iterations with a standard method that is complemented by a feature to fit in the current context. A further innovation is the computation of solutions in three-dimensional domains, which are still rare. Special attention is paid to applicability of the 3D simulation tools. The programs are designed to have justifiable working complexity. The simulation results of some models of contemporary semiconductor devices are shown and detailed comments on the results are given. Eventually, we make a prospect on future development and enhancements of the models and of the algorithms that we used.

Sociología y ética del deporte: Desde la perspectiva relacional

Esta tesis presenta un breve estado de la cuestión de la sociología y la ética del deporte como nueva disciplina académica, desde su origen hasta el presente. En segundo lugar pretende ver que aporta la perspectiva de la sociologia relacional al estudio del deporte. La primera contribución es la “trans-discipliniariedad” entre disciplinas como la sociología y la ética, evitando caer en una ética “sociologizzata” o en una sociología “eticizzata”. Gracias a la relacionalidad se obtiene la des-mercantilización de bienestar. La lógica relacional busca el desarrollo económico sin olvida que la sociedad está formada por personas, que son lo realmente importantes por encima de los interés económicos. Otra contribución es la distinción entre la “sociedad humana” y la “sociedad de lo humano” que evita muchos de los actuales problemas que trae consigo las nuevas tecnologías. Y finalmente aporta el esquema “AGIL” como “brújula relacional” que ayuda a relaizar los objetivos, medios y reglas éticas dentro de la práctica deportiva.

Social housing: interventions of densification in urban regeneration (1995-2010). Critical analysis of strategies adopted in Milan, London, São Paulo

The prospect of the continuous multiplication of life styles, the obsolescence of the traditional typological diagrams, the usability of spaces on different territorial scales, imposes on contemporary architecture the search for new models of living. Limited densities in urban development have produced the erosion of territory, the increase of the harmful emissions and energy consumption. High density housing cannot refuse the social emergency to ensure high quality and low cost dwellings, to a new people target: students, temporary workers, key workers, foreign, young couples without children, large families and, in general, people who carry out public services. Social housing strategies have become particularly relevant in regenerating high density urban outskirts. The choice of this research topic derives from the desire to deal with the recent accommodation emergency, according to different perspectives, with a view to give a contribution to the current literature, by proposing some tools for a correct design of the social housing, by ensuring good quality, cost-effective, and eco-sustainable solutions, from the concept phase, through management and maintenance, until the end of the building life cycle. The purpose of the thesis is defining a framework of guidelines that become effective instruments to be used in designing the social housing. They should also integrate the existing regulations and are mainly thought for those who work in this sector. They would aim at supporting students who have to cope with this particular residential theme, and also the users themselves. The scientific evidence of either the recent specialized literature or the solutions adopted in some case studies within the selected metropolitan areas of Milan, London and São Paulo, it is possible to identify the principles of this new design approach, in which the connection between typology, morphology and technology pursues the goal of a high living standard.

A Semantic-based Framework for Digital Survey of Architectural Heritage

The research aims at developing a framework for semantic-based digital survey of architectural heritage. Rooted in knowledge-based modeling which extracts mathematical constraints of geometry from architectural treatises, as-built information of architecture obtained from image-based modeling is integrated with the ideal model in BIM platform. The knowledge-based modeling transforms the geometry and parametric relation of architectural components from 2D printings to 3D digital models, and create large amount variations based on shape grammar in real time thanks to parametric modeling. It also provides prior knowledge for semantically segmenting unorganized survey data. The emergence of SfM (Structure from Motion) provides access to reconstruct large complex architectural scenes with high flexibility, low cost and full automation, but low reliability of metric accuracy. We solve this problem by combing photogrammetric approaches which consists of camera configuration, image enhancement, and bundle adjustment, etc. Experiments show the accuracy of image-based modeling following our workflow is comparable to that from range-based modeling. We also demonstrate positive results of our optimized approach in digital reconstruction of portico where low-texture-vault and dramatical transition of illumination bring huge difficulties in the workflow without optimization. Once the as-built model is obtained, it is integrated with the ideal model in BIM platform which allows multiple data enrichment. In spite of its promising prospect in AEC industry, BIM is developed with limited consideration of reverse-engineering from survey data. Besides representing the architectural heritage in parallel ways (ideal model and as-built model) and comparing their difference, we concern how to create as-built model in BIM software which is still an open area to be addressed. The research is supposed to be fundamental for research of architectural history, documentation and conservation of architectural heritage, and renovation of existing buildings.

Amphotericin B nano-formulations : development, characterisation and suitability for oral administration

In der Form von Nanokapseln (AmB-HST), Nanoemulsion beziehungsweise multilamellaren Vesikeln (MLV) wurden drei Amphotericin-B-Formulierungen für die orale Applikation entwickelt, charakterisiert und verglichen. Die neuartige homogene Nanokapsel-Formulierung des hydrophoben Polyen-Antimykotikums Amphotericin B wurde in Analogie zu einem für Simvastatin und andere Arzneistoffe etablierten Prozess aus der Reinsubstanz, Lezithin und Gelatine mit Hilfe des HST-Verfahrens hergestellt. Photometrische Untersuchungen zeigten, dass das Endprodukt aus Monomeren aufgebaut ist. Mittels Mikroskopie ließen sich die Aggregate vor der Umhüllung mit Lezithin und Gelatine im Ausgangsmaterial als individuelle kugelförmige Arzneistoffpartikel darstellen. Strukturuntersuchungen mit dynamischer licht streuung (DLS) zeigten eine enge Größenverteilung der verkapselten Partikel von ca. 1 µm. Die Struktur der Hülle der HST-Partikel wurde erstmalig mit Neutronenstreuung unter Verwendung der Deuterium-basierten Lösungsmittel kontrastmethode aufgeklärt. Durch die teilweise Kontrastmaskierung des Partikelkerns bei der Neutronenstreuung konnte die Lezithin-Gelatine-Hülle als eine dünne, 5,64 ± 0.18 nm dicke Schicht aufgelöst werden, welche der biologischen Lipidmembran ähnlich, im Vergleich aber geringfügig größer ist. Dieses Resultat eröffnet Wege für die Optimierung der Formulierung von pharmazeutischen Nanopartikeln, z.B. durch Oberflächenmodifizierungen. Weitere Untersuchungen mittels Kleinwinkelneutronenstreuung unter Verwendung der D-Kontrastvariation deuten darauf hin, dass die Komponenten der Nanokapseln nicht den gleichen Masseschwerpunkt haben, sondern asymmetrisch aufgebaut sind und dass die stärker streuenden Domänen weiter außen liegen. Die Partikel sind im Vergleich zu Liposomen dichter. In-Vitro Freisetzungsstudien belegen das Solubilisierungsvermögen des HST-Systems, wonach die Freisetzung des Arzneistoffes aus der Formulierung zu allen gemessenen Zeitpunkten höher als diejenige der Reinsubstanz war. rnDie Nanoemulsion-Formulierung von Amphotericin B wurde mit einem Öl und Tensid system, jedoch mit unterschiedlichen Co-Solvenzien, erfolgreich entwickelt. Gemäß der Bestimmung der Löslichkeit in verschiedenen Hilfsstoffen erwies sich der Arzneistoff Amphotericin B als nicht-lipophil, gleichzeitig aber auch als nicht-hydrophil. Die zur Ermittlung der für die Emulsionsbildung notwendigen Hilfstoffkonzentrationen erstellten ternären Diagramme veranschaulichten, dass hohe Öl- und Tensidgehalte zu keiner Emulsionsbildung führten. Dementsprechend betrug der höchste Ölgehalt 10%. Die Tröpfchengröße wuchs mit zunehmender Tensidkonzentration, wobei die Co-Solventmenge der Propylenglykol-haltigen Nanoemulsion indirekt verringert wurde. Für die Transcutol®P-haltige Nanoemulsion hingegen wurde das Gegenteil beobachtet, nämlich eine Abnahme der Tröpfchengröße bei steigenden Tensidkonzentrationen. Durch den Einschluss des Arzneistoffes wurde nicht die Viskosität der Formulierung, sondern die Tröpfchengröße beeinflusst. Der Wirkstoffeinschluss führte zu höheren Tröpfchengrößen. Mit zunehmender Propylenglykolkonzentration wurde der Wirkstoffgehalt erhöht, mit zunehmender Transcutol®P-Konzentration dagegen vermindert. UV/VIS-spektroskopische Analysen deuten darauf hin, dass in beiden Formulierungen Amphotericin B als Monomer vorliegt. Allerdings erwiesen sich die Formulierungen Caco-2-Zellen und humanen roten Blutkörperchen gegenüber als toxisch. Da die Kontrollproben eine höhere Toxizität als die wirkstoffhaltigen Formulierungen zeigten, ist die Toxizität nicht nur auf Amphotericin, sondern auch auf die Hilfsstoffe zurückzuführen. Die solubilisierte Wirkstoffmenge ist in beiden Formulierungen nicht ausreichend im Hinblick auf die eingesetzte Menge an Hilfsstoff nach WHO-Kriterien. Gemäß diesen Untersuchungen erscheinen die Emulsions-Formulierungen für die orale Gabe nicht geeignet. Dennoch sind Tierstudien notwendig, um den Effekt bei Tieren sowie die systemisch verfügbare Wirkstoffmenge zu ermitteln. Dies wird bestandskräftige Schlussfolgerungen bezüglich der Formulierung und Aussagen über mögliche Perspektiven erlauben. Nichtsdestotrotz sind die Präkonzentrate sehr stabil und können bei Raumtemperatur gelagert werden.rnDie multilamellar-vesikulären Formulierungen von Amphotericin B mit ungesättigten und gesättigten neutralen Phospholipiden und Cholesterin wurden erfolgreich entwickelt und enthielten nicht nur Vesikel, sondern auch zusätzliche Strukturen bei zunehmender Cholesterinkonzentration. Mittels Partikelgrößenanalyse wurden bei den Formulierungen mit gesättigten Lipiden Mikropartikel detektiert, was abhängig von der Alkylkettenlänge war. Mit dem ungesättigten Lipid (DOPC) konnten hingegen Nanopartikel mit hinreichender Verkapselung und Partikelgrößenverteilung gebildet werden. Die Ergebnisse der thermischen und FTIR-spektroskopischen Analyse, welche den Einfluss des Arzneistoffes ausschließen ließen, liefern den Nachweis für die mögliche, bereits in der Literatur beschriebene Einlagerung des Wirkstoffs in lipid- und/oder cholesterinreiche Membranen. Mit Hilfe eines linearen Saccharosedichtegradienten konnte die Formulierung in Vesikel und Wirkstoff-Lipid-Komplexe nach bimodaler Verteilung aufgetrennt werden, wobei der Arzneistoff stärker mit den Komplexen als mit den Vesikeln assoziiert ist. Bei den Kleinwinkelneutronenstreu-Experimenten wurde die Methode der Kontrastvariation mit Erfolg angewendet. Dabei konnte gezeigt werden, dass Cholesterol in situ einen Komplex mit Amphotericin B bildet. Diesen Sachverhalt legt unter anderem die beobachtete Differenz in der äquivalenten Streulängendichte der Wirkstoff-Lipid- und Wirkstoff-Lipid-Cholesterin-haltigen kleinen unilamellaren Vesikeln nahe. Das Vorkommen von Bragg-Peaks im Streuprofil weist auf Domänen hin und systematische Untersuchungen zeigten, dass die Anzahl der Domänen mit steigendem Cholesteringehalt zunimmt, ab einem bestimmten Grenzwert jedoch wieder abnimmt. Die Domänen treten vor allem nahe der Außenfläche der Modellmembran auf und bestätigen, dass der Wirkstoff in den Cholesterinreichen Membranen vertikal eingelagert ist. Die Formulierung war sowohl Caco-2-Zellen als auch humanen roten Blutkörperchen gegenüber nicht toxisch und erwies sich unter Berücksichtigung der Aufnahme in Caco-2-Zellen als vielversprechend für die orale Applikation. Die Formulierung zeigt sich somit aussichtsreich und könnte in Tabletten weiterverarbeitet werden. Ein Filmüberzug würde den Wirkstoff gegen die saure Umgebung im Magen schützen. Für die Bestimmung der systemischen Verfügbarkeit der Formulierung sind Tierstudien notwendig. Die entwickelten multilamellaren Formulierungen einschließlich der Wirkstoff-Cholesterin-Komplexe bieten somit gute Aussichten auf die mögliche medizinische Anwendung. rnrn

MRI of hip osteoarthritis and implications for surgery

Osteoarthritis of the hip joint is caused by a combination of intrinsic factors and extrinsic factors. Different surgical techniques are being performed to delay or halt osteoarthritis. Success of salvage procedures of the hip depends on the existing cartilage and joint damage before surgery; the likelihood of therapy failure rises with advanced osteoarthritis. For imaging of intra-articular hip pathology, MR imaging represents the best technique because of its ability to directly visualize cartilage, superior soft tissue contrast, and the prospect of multidimensional imaging. This article gives an overview on the standard MR imaging techniques used for diagnosis of hip osteoarthritis and their implications for surgery.

From nature versus nurture, via nature and nurture, to gene x environment interaction in mental disorders

It is now generally accepted that complex mental disorders are the results of interplay between genetic and environmental factors. This holds out the prospect that by studying G x E interplay we can explain individual variation in vulnerability and resilience to environmental hazards in the development of mental disorders. Furthermore studying G x E findings may give insights in neurobiological mechanisms of psychiatric disorder and so improve individualized treatment and potentially prevention. In this paper, we provide an overview of the state of field with regard to G x E in mental disorders. Strategies for G x E research are introduced. G x E findings from selected mental disorders with onset in childhood or adolescence are reviewed [such as depressive disorders, attention-deficit/hyperactivity disorder (ADHD), obesity, schizophrenia and substance use disorders]. Early seminal studies provided evidence for G x E in the pathogenesis of depression implicating 5-HTTLPR, and conduct problems implicating MAOA. Since then G x E effects have been seen across a wide range of mental disorders (e.g., ADHD, anxiety, schizophrenia, substance abuse disorder) implicating a wide range of measured genes and measured environments (e.g., pre-, peri- and postnatal influences of both a physical and a social nature). To date few of these G x E effects have been sufficiently replicated. Indeed meta-analyses have raised doubts about the robustness of even the most well studied findings. In future we need larger, sufficiently powered studies that include a detailed and sophisticated characterization of both phenotype and the environmental risk.

Assessing land use and land cover change in tropical dry forest or Northern Chinandega, Nicaragus from 1985 to 2011

The objective of this study is to gain a quantitative understanding of land use and land cover change (LULCC) that have occurred in a rural Nicaraguan municipality by analyzing Landsat 5 Thematic Mapper (TM) images. By comparing the potential extent of tropical dry forest (TDF) with Landsat 5 TM images, this study analyzes the loss of this forest type on a local level for the municipality of San Juan de Cinco Pinos (63.5 km2) in the Department of Chinandega. Change detection analysis shows where and how land use has changed from 1985 to the present. From 1985 to 2011, nearly 15% of the TDF in San Juan de Cinco Pinos was converted to other land uses. Of the 1434.2 ha of TDF that was present in 1985, 1223.64 ha remained in 2011. The deforestation is primarily a result of agricultural expansion and fuelwood extraction. If current rates of TDF deforestation continue, the municipality faces the prospect of losing its forest cover within the next few decades.

EFFECT OF ELECTRICAL DISCHARGE PATTERN ON SPARK IGNITED FLAME KERNEL DEVELOPMENT

Fuel-lean combustion and exhaust gas recirculation (EGR) in spark ignition engines improve engine efficiency and reduce emission. However, flame initiation becomes more difficult in lean and dilute fuel-air mixture with traditional spark discharge. This research proposal will first provide an intensive review on topics related to spark ignition including properties of electrical discharge, flame kernel behavior and spark ignition modeling and simulation. Focus will be laid on electrical discharge pattern effect as it is showing prospect in extending ignition limits in SI engines. An experimental setup has been built with an optically accessible constant volume combustion vessel. Multiple imaging techniques as well as spectroscopy will be applied. By varying spark discharge patterns, preliminary test results are available on consequent flame kernel development. In addition to experimental investigation of spark plasma and flame kernel development, spark ignition modeling with detailed description of plasma channel is also proposed for this study.

Toward an early diagnosis of lung cancer: an autoantibody signature for squamous cell lung carcinoma

Serum-based diagnosis offers the prospect of early lung carcinoma detection and of differentiation between benign and malignant nodules identified by CT. One major challenge toward a future blood-based diagnostic consists in showing that seroreactivity patterns allow for discriminating lung cancer patients not only from normal controls but also from patients with non-tumor lung pathologies. We addressed this question for squamous cell lung cancer, one of the most common lung tumor types. Using a panel of 82 phage-peptide clones, which express potential autoantigens, we performed serological spot assay. We screened 108 sera, including 39 sera from squamous cell lung cancer patients, 29 sera from patients with other non-tumor lung pathologies, and 40 sera from volunteers without known disease. To classify the serum groups, we employed the standard Naïve Bayesian method combined with a subset selection approach. We were able to separate squamous cell lung carcinoma and normal sera with an accuracy of 93%. Low-grade squamous cell lung carcinoma were separated from normal sera with an accuracy of 92.9%. We were able to distinguish squamous cell lung carcinoma from non-tumor lung pathologies with an accuracy of 83%. Three phage-peptide clones with sequence homology to ROCK1, PRKCB1 and KIAA0376 reacted with more than 15% of the cancer sera, but neither with normal nor with non-tumor lung pathology sera. Our study demonstrates that seroreactivity profiles combined with statistical classification methods have great potential for discriminating patients with squamous cell lung carcinoma not only from normal controls but also from patients with non-tumor lung pathologies.