Dissertations / Theses on the topic 'DDAI'

Questa ricerca analizza gli studi che identificano un numero rilevante di bambini iperattivi nell‘Istruzione infantile della Rete Comunale delle Scuole di Porto Alegre. L‘enunciato scolastico produce una diagnosi e indirizza ai medici, ma con quale intento? Che cosa cambia nell‘organizzazione pedagogica a partire dalla diagnosi o dalla perizia medica? Che cosa segnala il corpo? Che cosa preannuncia il corpo? Le domande sono state tradotte in modo da organizzare il punto centrale dello studio: Che cosa accade con il sapere e il non sapere dell'istruzione rispetto ai Corpi Che Non Si Fermano? Per dare supporto alle ricerche, sono stati sviluppati alcuni concetti fondamentali. Il concetto di normalità, sulla base del pensiero di Canguilhem e Foucault, si associa al pensiero sistemico di Gregory Bateson. Considerando la continua articolazione tra normale e anormale, si cerca la costruzione del concetto di Disturbo da Deficit di Attenzione da Iperattività – DDAI, basandosi su riferimenti storici di costruzione di questi concetti e le evidenze per cui la diagnosi relativa a tali soggetti si presenta sottoforma di epidemia nel contesto analizzato. Un‘analisi cartografica dell‘attenzione, concetto anch‘esso centrale nella tesi, mette in evidenza l‘Attenzione intesa non come condizione preesistente, ma nel suo movimento circolare di invenzione, come effetto dell‘/nell‘apprendimento. Il lavoro di ricerca è stato sviluppato in particolare su bebè e bambini piccoli che integrano la Rete Scolastica Comunale di Porto Alegre, attraverso l‘attenzione ai discorsi degli Educatori Speciali che si prendono cura di questi bambini e prestano assistenza alle scuole. A seguito dei colloqui si è formata la composizione di gruppi di argomentazione che, sottoforma di piccoli titoli, vengono chiamati Nomi-Domande: Quando i bambini si fermano? Come ci si riferisce a questi bambini? Quali parole, che gruppi di argomentazione sono utilizzati? In che modo lo stato di abbandono si manifesta nei bambini, nelle famiglie e nei professori? Tra le evidenze, una delle più importanti è quella che identifica l‘iperattività associata a una dimensione di abbandono diretta non solo al soggetto bambino, ma a tutti i personaggi implicati nella rete. L‘abbandono, inteso come una forma acuta di sofferenza di diverso ordine finisce per produrre un‘esistenza di sofferenza. Tuttavia, a partire dal punto di vista diretto al contesto, si è osservato che i bambini in stato di abbandono, quando hanno ricevuto cure e attenzione personalizzate, hanno presentato un cambiamento che si traduce in possibilità di vita in comune e apprendimento nella scuola.
Esta pesquisa analisa os discursos que identificam um número expressivo de crianças como hiperativas na Educação Infantil da Rede Municipal de Ensino de Porto Alegre. O discurso escolar produz diagnóstico e encaminha aos consultórios médicos, mas com que intenção? O que muda na organização pedagógica a partir do diagnóstico ou do laudo médico? O que denuncia o corpo? O que anuncia o corpo? As perguntas foram traduzidas de forma a organizar a questão central da pesquisa: O que acontece com o saber e o não saber da educação face aos Corpos Que Não Param? Para dar sustentação às buscas, foram desenvolvidos alguns conceitos fundamentais. O conceito de normalidade, tendo como base o pensamento de Canguilhem e Foucault, associa-se ao pensamento sistêmico de Gregory Bateson. Considerando a contínua articulação entre normal e anormal busca-se a construção do conceito de Transtorno de Déficit de Atenção com Hiperatividade – TDAH, tomando as referências históricas de construção desse conceito e as evidências de que o diagnóstico referente a tais sujeitos se apresenta na forma de epidemia no contexto investigado. Uma análise cartográfica da atenção, conceito também central na tese, dá evidência à Atenção entendida não como condição prévia, mas em seu movimento circular de invenção, como efeito da/na aprendizagem. O trabalho de investigação foi desenvolvido conferindo destaque a bebês e crianças pequenas que integram a Rede Municipal de Ensino de Porto Alegre, por meio da atenção aos discursos das Educadoras Especiais que prestam atendimento a essas crianças e assessoria às escolas. A partir das entrevistas, houve a composição de grupos de argumentação que, em forma de pequenos títulos, são chamados de Nomes-Perguntas: Quando as crianças param? Como essas crianças são referidas? Que palavras, que grupos de argumentações são usados? Como o desamparo se mostra nas crianças, famílias e professoras? Dentre as evidências, uma das mais importantes é a que identifica hiperatividade associada a uma dimensão de desamparo, dirigida não só ao sujeito criança, mas a todos os implicados nessa rede. O desamparo, entendido como uma forma aguda de sofrimento de diversas ordens acaba por produzir uma existência de sofrimento. No entanto, a partir do olhar dirigido ao contexto, foi observado que crianças em estado de desamparo, quando encontraram o cuidado e a atenção personalizada, apresentaram uma mudança que se traduz em possibilidade de convivência e aprendizagem na escola.
This research analyzes the several discourses that identify a significant number of children as hyperactive in Child Education in the Schools of the City of Porto Alegre. The school discourse results in diagnosis and refers to doctor's offices, but with what purpose? What changes in educational organization with the diagnosis or medical report? What does the body denounce? What does the body announce? These questions were translated into a way to organize the focal point of the research: What happens with knowing and not knowing in education vis-à-vis Bodies That Will Not Stop? In order to substantiate the searches, some essential concepts were developed. The concept of normalcy, based on the thinking of Canguilhem and Foucault, is associated to the systemic thinking of Gregory Bateson. Considering the continuous articulation between normal and abnormal, the construction of the concept of Attention Deficit Hyperactivity Disorder - ADHD is pursued, using the historical references to build this concept and the evidence that the diagnosis for such subjects takes the form of epidemics within the investigated context. A cartographic analysis of attention, also an essential concept in this paper, evidences understood Attention not as a condition precedent, but within its circular invention movement, as an effect of and in learning. The investigation work was conducted with focus on babies and small children of the Schools in the City of Porto Alegre, by means of attention to the discourses of Special Educators tending to these children and assisting these schools. Based on the interviews, groups of arguments were formed under small titles, called Question-Names: When do the children stop? How are these children referred? Which words, which groups of arguments are used? How does abandonment show in children, families and teachers? Among the evidences, one of the most important is the one that identifies hyperactivity associated to a dimension of abandonment, focused not only on the subject child but also on all of those affected in this network. Abandonment, understood as an acute form of suffering of several orders, ultimately produces a painful existence. However, by focusing on the context, it was observed that children in an abandonment state, when experiencing personal care and attention, showed a change that translates into the possibility of interacting and learning at school.
Esta investigación analiza los discursos que identifican un número expresivo de niños como hiperactivos en la Educación Infantil de la Red Municipal de Enseñanza de Porto Alegre. El discurso escolar produce diagnóstico y encamina a los consultorios médicos, pero, ¿con qué intención? ¿Qué cambia en la organización pedagógica a partir del diagnóstico o del laudo médico? ¿Qué denuncia el cuerpo? ¿Qué denuncia el cuerpo? Las preguntas se tradujeron para organizar la cuestión central de la investigación: ¿Qué ocurre con el saber y el no saber de la educación de cara a los Cuerpos Que No Paran? Para dar sustentación a las búsquedas, se desarrollaron algunos conceptos fundamentales. El concepto de normalidad, teniendo como base el pensamiento de Canguilhem y Foucault, se asocia al pensamiento sistémico de Gregory Bateson. Considerando la continua articulación entre normal y anormal se busca la construcción del concepto de Trastorno de Déficit de Atención con Hiperactividad – TDAH, tomando las referencias históricas de construcción de ese concepto y las evidencias de que el diagnóstico referente a tales sujetos se presenta en la forma de epidemia en el contexto investigado. Un análisis cartográfico de la atención, concepto también central en la tesis, da evidencia a la Atención entendida no como condición previa, sino en su movimiento circular de invención, como efecto del/en el aprendizaje. El trabajo de investigación fue desarrollado dándole destaque a bebés y a niños pequeños que integran la Red Municipal de Enseñanza de Porto Alegre, por medio de la atención a los discursos de las Educadoras Especiales que prestan atención a esos niños y asesoría a las escuelas. A partir de las entrevistas, hubo la composición de grupos de argumentación que, en forma de pequeños títulos, son llamados de Nombres-Preguntas: ¿Cuándo paran los niños? ¿Cómo se refieren a esos niños? ¿Qué palabras, qué grupos de argumentaciones se usan? ¿Cómo se muestra el desamparo en los niños, familias y profesoras? Entre las evidencias, una de las más importantes es la que identifica hiperactividad asociada a una dimensión de desamparo, dirigida no sólo al sujeto niño, sino a todos los implicados en esa red. El desamparo, entendido como una forma aguda de sufrimiento de diversos órdenes acaba por producir una existencia de sufrimiento. Sin embargo, a partir de la mirada dirigida al contexto, se observó que niños en estado de desamparo, cuando encontraron el cuidado y la atención personalizada, presentaron un cambio que se traduce en posibilidad de convivencia y aprendizaje en la escuela.

Nitric oxide (NO) is a key physiological messenger, but an excessive production of this molecule can be detrimental, leading to the onset or worsening of many pathological conditions. Dimethylarginine dimethylaminohydrolase (DDAH) is a key enzyme in the NO pathway, involved in the metabolism of asymmetric dimethylarginine (ADMA) and monomethyl arginine (L-NMMA), which are both endogenous inhibitors of NO synthesis. Two isoforms of DDAH have been identified in humans, namely DDAH-1 and DDAH-2. DDAH inhibition represents a promising strategy in the treatment of NO overproduction under pathological conditions without affecting the homeostatic role of this messenger. In this work I described the design and synthesis of 12 novel potential DDAH inhibitors together with the development of a new UPLC-MS based assay to measure the activity of HEK293T cell lysates overexpressing recombinant human DDAH-1 in metabolizing ADMA into dimethylamine and L-citrulline. The same assay was used to assess the potential of the novel compounds, as well as of the well-known DDAH inhibitor L-257, to inhibit DDAH-1 catalyzed L-citrulline formation from ADMA. Three of the novel molecules (compounds 10a, 14a and 14b) showed very interesting inhibitory activity: in particular, the methylacylsulfonamide analogue of L-257 (10a) resulted in 13-fold higher inhibition potency than L-257 itself (98% of inhibition at 1mM, IC50 = 3±3 μM and Ki = 1±0 μM). This molecule was chosen for molecular dynamics simulations to study the putative mechanism for 10a inhibition of DDAH-1 activity. Furthermore, DDAH-1 and DDAH-2 were engineered introducing a FLAG-tag at the C-terminal of the proteins to allow their purification from the lysate components by immunoprecipitation. Although the purification protocol requires some further improvement, the fusion proteins did not show to be functionally affected by the modification.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Dias da Silva, Tarciana; Fawzi Hadj Sadok, Djamel. DDAN: A distributed directory for ambient networks. 2008. Dissertação (Mestrado). Programa de Pós-Graduação em Ciência da Computação, Universidade Federal de Pernambuco, Recife, 2008.

Distributed digital arrays have many potential applications in radar and communication systems. The objective of this thesis is to re-examine previous research on distributed digital array radar (DDAR) and evaluate several critical aspects of a proposed wireless architecture. Self-standing transmit/receive (T/R) modules are synchronized wirelessly. An important issue addressed in this thesis is whether a simple low-cost synchronization circuit would perform adequately. To this end two breadboard T/R modules were built to support test and evaluation. Both measurements and simulations were performed. Other issues addressed in the research include a comprehensive investigation of the demodulator performance, and the development of Controller and processing software in LabVIEW.

Thesis (M.S. in Electronic Warfare Systems Engineering)--Naval Postgraduate School, September 2008.
Thesis Advisor(s): Jenn, David. "September 2008." Description based on title screen as viewed on November 3, 2008. Includes bibliographical references (p. 153-155). Also available in print.

In 2008, the Canadian standard for decking installed above ground was revised, and the penetration requirement was eliminated. This decision was based on field test data and fundamental work on mobility of copper in the preservative formulations that dominated the market at the time. Recently the wood protection industry has shown interest in shifting towards carbon-based preservatives. Thus, it is important to test the efficacy of carbon-based preservative formulations as shell-treatments on Canadian wood species. In this study, samples of spruce heartwood were treated with a formulation containing either tebuconazole or didecyldimethylammonium carbonate (DDAC). The treated wood was exposed outdoors for one year and the leachate from these samples was collected. Tebuconazole and DDAC were detected in the leachate collected, and this indicates that these active ingredients were mobile in the wood after treatment. DDAC was detected in very low concentrations on wood surfaces that were untreated before exposure: 0.03 mg DDAC/g of wood and 0.02 mg of DDAC/g wood were measured for the high and low retentions respectively. The concentration of tebuconazole detected was not different from the control samples. This suggests that mobile DDAC may be able to re-deposit in the wood, but tebuconazole does not re-deposit once it is dislodged from the wood. Furthermore, results showed that spores of Gloeophyllum sepiarium and Oligoporus placentus were able to germinate on untreated check surfaces within 2 weeks on samples collected from exposed, treated wood. This indicates that the re-deposited carbon-based active ingredients were not able to protect the untreated check surfaces against germination of basidiospores of some common fungi isolated from above-ground decking in Canada.

Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide (NO) synthesis and elevated plasma levels associate with poor cardiovascular and renal outcomes. The dimethylarginine dimethylaminohydrolase enzymes (DDAHs; 1 and 2) metabolise ADMA. A DDAH1 gene variant associates with higher kidney tissue mRNA expression, lower plasma ADMA but counter-intuitively, a steeper rate of eGFR decline. This indicates that renal DDAH1 activity may be deleterious and circulating ADMA does not necessarily reflect the NO-ADMA balance (or severity of disease) within kidney tissue. This study tests the hypothesis that reduced renal DDAH1 activity protects against the progression of kidney function decline, independent of circulating ADMA. Renal DDAH1 expression predominates within the proximal tubule. A novel proximal tubule-specific DDAH1 knock-out (PTD1KO) mouse was developed, which demonstrated tubule-specific dysregulation of ADMA and NO that was not evident systemically. Phenotyping studies in PTD1KO mice did not identify consistent alterations of urinary biochemistry at baseline or after salt loading, however, proteomic analysis revealed significant alterations of urinary peptides at baseline; including down-regulation of uromodulin and collagen. At 12 weeks following folate renal injury, the PTD1KO mouse exhibited less kidney function decline, collagen deposition and pro-fibrotic gene expression (Col12alpha, TGFbeta and ET-1) than controls. Furthermore, ADMA and DDAH1 inhibition reduced tubular sodium and fluid reabsorption in rat microperfusion studies, although studies in PTD1KO mice failed to reproduce this effect. Finally, in vitro studies using a PT cell line and primary PT culture indicated an inhibitory effect of ADMA upon PT cell proliferation. Consistent with recent human genetic studies, these data provide experimental evidence indicating a reduction of renal tubule DDAH1 activity can protect against progressive kidney fibrosis and function decline, independent of plasma ADMA. This work provides novel insights into the role of the NO-ADMA-DDAH axis within the kidney, particularly the tubule.

Genom laborativa försök skulle avvattning och retention av nanocellulosa i en DDA (Dynamic drainage analyzer) undersökas. Detta genom att tillsätta retentionskemikalier i varierande mängder för att se hur det påverkade avvattningen. Uppdragsgivaren var Innventia och laborationerna utfördes på SP:s laboratorium där DDA instrument fanns tillgängligt. DDA instrumentet liknar den maskin som används för papperstillverkning och därför anses det vara möjligt att kunna avvattna nanocellulosa på liknande instrument. Målet med examensarbetet var att få fram en nanocellulosafilm med goda barriärsegenskaper men också se hur tillsats av bärarfibrer påverkar filmens egenskaper. Målet var också att se om retention och avvattningstiden för nanocellulosa hänger ihop och om det går att få fram repeterbara resultat. Två olika viror undersöktes också i de laborativa experimenten i DDA, en som benämns som Albanyviran och en som benämns som Stratexviran. Albanyviran är tätare än Stratexviran och effekten av viratäthet på retention och avvattningstid undersöktes. Nanocellulosa eller Mikrofibrillär cellulosa (MFC) är ett nytt och förnybart material som utvinns ur träfibrer och karakteriseras av sitt geléaktiga utseende[4]. Nanocellulosan lämpar sig utmärkt för en mängd olika produkter t.ex. som barriär, enskilt i form av filmer eller i blandade produkter. Vid tillverkning av nanocellulosa används en homogenisator som sönderdelar cellulosafibrer till fibriller och fibrillaggregat. Detta var tidigare ett problem då fibrerna satte igen homogenisatorn och tillverkningen var mycket energikrävande[4]. När det gäller filmtillverkning av nanocellulosa kvarstår problemet när det kommer till avvattningen. Nanocellulosan späddes till önskad koncentration och innan de laborativa försöken kördes den genom en homogenisator för att dispergera fibrillerna i vätskan efter spädningen. Bärarfibrerna slogs upp i en uppslagare med två liter kranvatten för att sedan tillsättas till den homogeniserade nanocellulosan. Under försöket i DDA varierades mängden och andelen MFC (mikrofibrillär cellulosa) och bärarfibrer (Modorefmassa). Till MFC och bärarfibersuspensionen i DDA:n tillsattes sedan två retentionskemikalier vid varje försök, C-PAM PL-1520 och EKA NP-780 i varierande mängder. Efter avvattningen i DDA:n pressades filmen vid olika tryck och tider för att därefter mäta filmernas ytvikt och syrgasbarriär. Den film som ansågs mest lämplig gällande avvattning i DDA under försöken var vid 0,2 % med 90 % MFC och 10 % bärarfibrer. Filmen gav den högsta retentionen, en god syrgasbarriär och var lätt att hantera. Det som kan ses från resultaten av syrgasbarriären är att vid 0,2 % med 90 % MFC och 10 % bärarfibrer erhölls det lägsta OTR (oxygen transmission rate) -värdet på 0,53 vilket visar på en bra syrgasbarriär. Retentionen för det försöket var det högsta på 87,1 % medan avvattningstiden låg på närmare 250 sekunder. Avvattningstiderna var höga, dock så var det vid denna totalkoncentration ibland svårt att se när avvattningen avslutades då tiden klockades manuellt. Det som kan ses utifrån resultaten är att bärarfibrerna inte påverkar filmerna negativt utan kan gynna både så retentionen och syrgasbarriären blir bättre, dock fås en högre ytvikt och avvattningstiderna blir längre.
Through laboratory experiments, dewatering and retention of nanocellulose in a DDA (Dynamic Drainage Analyzer) were analysed. By adding retention chemicals in varied amounts, the effects on the dewatering was shown. The Job initiator was Innventia and the laboratory work were made at SP's laboratory where the DDA instrument was available. The DDA instrument is similar to the machine that is used for paper manufacturing in a large scale and therefore, it could be possible to dewater nanocellulose on a similar instrument. The goal of the thesis was to develop a nanocellulose film with good barrier properties but also to see how adding carrier fibers effect the properties of the film. The goal was also to see if the retention and dewatering time of nanocellulose are connected and whether it is possible to obtain repeatable results. Two different wires were also examined in the laboratory experiments in a DDA, the Albanywire and the Stratexwire. The Albany wire was denser than the Stratex wire and the effect that the density caused on retention and dewatering time was examined. Nanocellulose or Microfibrillated cellulose (MFC) is a new and renewable material that is made from wood fibers and is characterized by its gelatinous appearance. [4] Nanocellulose is suited for a variety of products, such as barriers, alone in the form of films or mixed in products. In the manufacture of nanocellulose a homogenizer is used which decomposes cellulose fibers to fibrils fibril aggregate. This was previously a problem while the fibers clogged the homogenizer and the production had a high energy consumption. [4] When it comes to making a nanocellulose film the problems with dewatering remains. The nanocellulose was diluted to the desired concentration and before the laboratory experiments it was run through a homogenizer, to disperse the fibrils in the liquid after the dilution. The carrier fibers was prepared in a blender with two liters of tap water before it was added to the homogenized nanocellulose. During the experiment in the DDA the amount and proportion of the MFC (microfibrillar cellulose) and carrier fibers (Modorefmassa) was varied. To the MFC and carrier fiber suspension in the DDA two retention chemicals were added in each experiment, C- PAM PL -1520 and EKA NP- 780 in varying amounts. After the dewatering of nanocellulose in the DDA the films were pressed at different pressures and times, thereafter the oxygen permeability was analyzed. The film that was considered the most suitable referring to dewatering in the DDA during the attempts was at 0.2 % with 90 % MFC and 10 % carrier fibers. The film gave the highest retention, a good oxygen barrier and was easy to handle. What can be seen from the results of the oxygen barrier measurement is that at 0.2 % with 90 % MFC and 10% carrier fibers obtained the lowest value OTR (oxygen transmission rate), which indicates on a good oxygen barrier. Retention at this concentration was the highest at 87.1 %, while the drainage time was nearly 250 seconds. The dewatering time was high, however during this concentration it’s sometimes difficult to see when the dewatering ended while the time was clocked manually. Conclusions from the results are that the carrier fibers doesn’t have a negatively effect on the films, rather they can benefit both the retention and oxygen barrier, however a higher paper weight was obtained and the dewatering time became longer.

The work deals with the analog design of low-voltage and low-power differential difference amplifier DDA with adaptive differential input stage, second stage class AB, improved by a self – cascode to achieve higher gain and slew rate. Conventional and unconventional techniques, and methods for low-voltage and low-power design are presented. The finished design of the differential difference amplifier DDA with the analyzed results is presented. Design of a Butterwortho low-pass filter of the sixth order based on DDA with Sallen Key topology and design of a multifunctional ARC filter based on DDA.

Mestrado em Engenharia Civil
A aplicação de geossintéticos, com diversas funções, em obras de Engenharia Civil é cada vez mais frequente, devido essencialmente às vantagens que apresentam em relação a outros materiais e tecnologias tradicionais. Nas estruturas ferroviárias a aplicação de geossintéticos tem-se revelado uma solução técnica e economicamente atractiva, sobretudo no caso de obras de reabilitação, devido ao aumento das velocidades de circulação e das cargas por eixo transportadas, e aos maiores níveis de segurança exigidos. Neste trabalho é feita uma contribuição para o conhecimento dos efeitos da danificação durante a instalação (DDI) e da abrasão em geossintéticos utilizados em obras ferroviárias, em particular, no seu comportamento mecânico de curto prazo (tracção). No Capítulo 1 é feita a apresentação da estrutura da dissertação. No Capítulo 2 é feita uma introdução geral aos geossintéticos, onde são enumeradas as matérias-primas mais utilizadas no seu fabrico, as principais propriedades e funções destes materiais, e ainda os principais agentes e mecanismos de degradação que afectam a durabilidade dos geossintéticos. O Capítulo 3 diz respeito à aplicação de geossintéticos em obras ferroviárias. Neste capítulo são apresentados os principais elementos que constituem as vias-férreas tradicionais (balastradas); é feita uma revisão bibliográfica onde são referidos alguns estudos relativos ao desempenho de geossintéticos em obras ferroviárias, nomeadamente no desempenho das funções de separação, filtragem/drenagem e reforço. Na parte final do capítulo são referidas as questões relativas à abrasão e danificação induzida durante a instalação em obra (DDI) de geossintéticos, onde são apresentados alguns estudos realizados por outros autores para quantificar os efeitos dos referidos fenómenos no comportamento dos geossintéticos. No Capítulo 4 é apresentado o estudo laboratorial desenvolvido para avaliar o comportamento a curto prazo dos geossintéticos após simulação dos fenómenos de abrasão e DDI; é feita a análise dos resultados obtidos e são apresentadas as principais conclusões do estudo. Por último, são apresentadas algumas sugestões para o desenvolvimento de futuras investigações.
The application of geosynthetics with different functions in works of Civil Engineering is increasingly common, due mostly to the advantages they offer over other materials and traditional technologies. The application of geosynthetics in railway structures has shown to be an attractive and economically technical solution, especially for rehabilitation works, due to the increase of the speeds of circulation and loads for axle carried, and to the higher levels of security required. In this work a contribution to the knowledge of the effects of the damage during installation (DDI) and abrasion of geosynthetics used in railways is made, particularly, in what concerns their short-term mechanical behaviour (tensile). In Chapter 1 the presentation of the structure of the dissertation is made. In Chapter 2 a general introduction to geosynthetics is made, where the raw materials used in their production, the main properties and functions of these materials and the most important agents and degradation mechanisms which affect the geosynthetics durability are presented. Chapter 3 concerns the application of geosynthetics in railway structures. The main elements that constitute the traditional railways (ballasted) are presented; a literature review which contains references to some studies on the performance of geosynthetics in railways, particularly in the performance of separation, filtration/drainage and reinforcement functions. At the end of the chapter the issues relating to the abrasion and damage induced during the installation (DDI) of geosynthetics in construction are referred, where some studies from other authors used to quantify the effects of such phenomena in the behaviour of geosynthetics are briefly described. In Chapter 4 the laboratory study carried out to assess the short-term behaviour of geosynthetics after simulation of the abrasion and DDI, the analysis of the results and the main conclusions of the study are presented. Finally, some suggestions for the development of future investigations are put forward.

El mètode DDCI (Difference Dedicated Configuration Interaction) s'ha aplicat sistemàticament a l'estudi de diferents problemes químics per a calcular de manera teòrica diferències d'energia entre estats. S'ha començat amb sistemes més simples per acabar aplicant el mètode a problemes de reactivitat. Malgrat l'extensa aplicació d'aquest mètode, mai havia estat emprat per a l'estudi de reaccions que impliquessin estats excitats.
En primer lloc, dins dels sistemes estàtics estudiats, s'ha determinat la multiplicitat de l'estat fonamental del biradical tetrametilenetà i s'ha analizat la influència dels substituents en l'estabilitat de l'estat triplet del carbè 2,6-dibromo-4-tert-butil-2',6'-bistrifluorometil-4'-isopropildifenilmetilè. Referent als problemes de reactivitat que impliquen estats excitats, s'ha recalculat la controvertida energia d'activació de l'etapa determinant de la velocitat de la reacció de descomposició quimiluminiscent del 1,2-dioxetà. Amb aquest estudi, el mètode DDCI ha demostrat donar resultats de qualitat similar o superior als d'altres mètodes multireferencials d'alt nivell convencionals (per exemple el mètode CASPT2) amb menor cost computacional. També s'ha estudiat la reacció de desactivació que té lloc en els àcids nucleics com a conseqüència de la irradiació per llum UV en el ADN. En concret, s'han estudiat els derivats de la citosina, 5-metilcitosina i 5-fluorocitosina utilitzant el mètode DDCI i d'altres estratègies computacionals, revelant en tots els casos els resultats la gran complexitat del mecanisme.
Amb aquest treball ha estat possible establir el ventall d'aplicabilitat del mètode i els paràmetres òptims per a la seva aplicació en cada cas.
The DDCI method (Difference Dedicated Configuration Interaction) has been systematically applied to study a variety of chemical problems calculating theoretical energy differences between states. We have begun with simpler static systems to end with reactivity problems. Despite the vast application of this method, it had never been applied to the study of reactions involving excited states.
Namely, in static systems we have determined the ground state multiplicity of the tetramethylenethane diradical and we have analysed the influence of the substituents on the triplet stability of the 2,6-dibromo-4-tert-buthyl-2',6'-bistrifluoromethyl-4'-isoprophyldiphenylmethylene. Concerning reactivity problems involving excited states, we have recalculated the controversial activation energy of the rate determining step of the chemiluminiscent decomposition of the 1,2-dioxetane. With this study the DDCI method has been shown to give results of the same or better quality than other multireference conventional methods (for instance CASPT2 method) with lower computational cost. Also, we have studied the deactivation reaction that takes place in nucleic acids as a result from the irradiation by UV light to the DNA. In particular, cytosine derivatives 5-methylcytosine and 5-fluorocytosine have been studied using the DDCI method among other computational strategies and the results have revealed the complexity of the mechanism.
With this work it has been possible to establish the range of applicability of this method and the optimal parameters for its application in each case.

This dissertation explores student environmentalism in Beijing, China. It traces students' political norms and values, explains their activism and experience of pollution, and investigates the role of environmental non-governmental organisation (ENGOs) in forming youth environmentalism. To serve these objectives, the work takes forward theories on youth activism and agency and recent debates on environmental health, environmentalism and ENGOs. This study was designed as a qualitative research project based primarily on interviews and complemented by ethnographic methods, content analysis, pictorial evidence and survey results provided by the Jane Goodall Institute China. Research findings and methodology are presented in four papers and a framing document. My work challenges labels of agency and activism as either protest and resistance or 'quiescence' and questions the influence of globalisation on activists' norms and values. I put forward 'fragmented activism' as a new concept to capture the nature of youth environmental activism in Beijing. I contribute to environmental health literature by tracing how young people develop discursive mechanisms to mitigate the fear of air pollution and argue that their response offers invaluable insights into the interplay between space and the body in polluted environments. This thesis further shows that the repertoire of student environmental associations in Beijing represents a type of 'place based environmentalism' (Smith, 2001) but argues that, whilst this may be a contradictory response to contemporary environmental issues, it is not usefully assessed against abstract and normative notions of what environmentalism should be. I also challenge scholarly assessments of ENGO action. By exploring ENGO strategies in China that rely on extant societal and governmental narratives about good citizenship and moral values - instead of radical alternatives to mainstream development models or political processes - I argue for new research paradigms guiding the study of environmental movements.

Includes bibliography.
Discontinuous Deformation Analysis (DDA) is a discrete element method developed by Shi [1988] specifically for modelling blocky rock masses. The DDA method is based on the assumption that deformation and failure of such rock masses is primarily due to differential movements of rock blocks, rather than strain and fracture of intact rock material. Strains and stresses are assumed to be constant over the area of each rock block. Contact between blocks is modelled using penalty functions, with Coulomb's friction law controlling sliding along block boundaries. Tests show that while DDA is not well suited to dynamic simulations where the velocities of blocks become large, it can model rock masses to a reasonable degree of accuracy in static analyses. There are various analysis control parameters which have a marked effect on the solution, however, and the user should take care in choosing suitable values for these parameters. A method is proposed here, in which certain blocks can be sub-divided into Finite Element meshes in order to obtain a more accurate description of their deformation. The method takes advantage of the fact that both DDA and the Finite Element Method (FEM) use the principle of stationary potential energy to obtain the solution equations for block equilibrium. Both DDA blocks and FEM elements can therefore initially be treated as DDA blocks, using the standard DDA formulation, and then the solution equations for the FEM elements are converted into Finite Element format by a simple transformation procedure before solution. First and second order DDA blocks are considered in this report, along with their equivalents in FEM, the C0-linear and C0-quadratic triangular elements. The C0-linear elements are found to be too stiff in modelling bending deformation, due to the assumption of constant strain throughout the element. The C0-quadratic elements are able to accurately model bending, however. It is shown through tests that the performance of these FEM elements, formulated within the DDA method, is identical to that obtained using the corresponding elements in conventional Finite Element programs. The sub-meshing method therefore allows mixed-formulation analyses, with DDA blocks and FEM meshes interacting within a single system, while remaining efficient, and reasonably simple to incorporate into existing DDA program codes. It would also be possible to model material non-linearity and fracture using this method.

O objetivo deste trabalho é verificar a aplicação do Dynamic Design Analysis Method DDAM na análise de estruturas submetidas a carregamentos de choque. Esse método foi desenvolvido pela Marinha Norte Americana e se propõe a estimar esforços inercias provenientes das respostas de equipamentos de bordo, sejam de navios ou submarinos, a cargas impulsivas, mais precisamente, ondas de choque submarinas. Para a compreensão plena dessa metodologia, foi necessário um estudo sobre vibrações e espectros de resposta, tópicos teóricos que fundamentam esse procedimento. Devido à escassez de informações divulgadas por se tratar de conteúdo militar, foi realizado um estudo de caso para uma estrutura simples, uma viga engastada e seus resultados foram comparados com um prévio trabalho publicado. Com isso, foi possível a aplicação do DDAM em uma estrutura mais complexa como o vaso do reator nuclear do tipo PWR de um submarino, ainda que em um modelo simplificado desse equipamento. No entanto, os resultados mostraram a validade do uso do DDAM na análise estrutural dinâmica de equipamentos de uma maneira qualitativa.
The main purpose of this dissertation is to verify the use of the Dynamic Design Analysis Method DDAM in the study of structures subjected to shock loadings. This method was developed by the U.S. Navy and is intended to estimate inertial forces resultant from the response of shipboard equipment, from surface ships or submarines, due to impulsive loads or, more precisely, underwater pressure shock waves. To fully comprehend this method, it was necessary to acknowledge fundamentals of dynamics of structures like free vibrations, response spectrum analysis correlated topics. And due to lack of public information since it is a military technology, a case study of a cantilever beam was done with its results compared to a previous published paper. Then, the DDAM was applied to a more complex structure a PWR nuclear reactor vase simplified model, in order to show the effectiveness of the method in dynamic structural analysis.

Alkyl dimethyl benzyl ammonium chloride (ADBAC) and didecyl dimethyl ammonium chloride (DDAC) are common quaternary ammonium compounds used as disinfectants in households, medical, and restaurant settings. They cause occupational skin and respiratory hazards in humans, and developmental and reproductive toxicity in mice. They also cause increased secretions of proinflammatory cytokines in cell lines and vaginal inflammation in porcine models; but have not been evaluated for developmental immunotoxicity. We assessed immunotoxicity in-vitro with J774A.1 murine macrophage cell line by analyzing cytokine production and phagocytosis; and evaluated developmental immunotoxicity in CD-1 mice by analyzing antibody production. Additionally, because of the associations between gut microbiome dysbiosis and immune disease, we monitored changes in the microbiome as a result of ADBAC+DDAC exposure. Production of cytokines TNF-alpha and IL-6 increased at low ADBAC+DDAC concentrations, and IL-10 decreased in the murine macrophages with ADBAC+DDAC exposure. The phagocytic function of macrophages was also severely decreased. ADBAC+DDAC altered the mouse microbiome by decreasing the relative abundance of Bacteroides and increases in Clostridia in F0 and F1 generations. IgG primary and secondary responses were altered in F1 male mice; and IgA and IgM production were decreased in secondary response in F2 male mice. Since ADBAC+DDAC show signs of immunotoxicity in mice, further studies are needed to reassess risk for human exposure as ADBAC+DDAC may be contributing to immune disease.
Master of Science

Mestrado em Engenharia Civil
Nos nossos dias, os resíduos municipais, na impossibilidade da sua reciclagem, encontram como último destino os aterros de resíduos sólidos urbanos (RSU). Estes resíduos podem conter substâncias que, caso não sejam devidamente isoladas do ambiente envolvente, podem levar à sua contaminação. Ao longo dos últimos anos os geossintéticos traduziram-se numa boa aposta na impermeabilização dos aterros de RSU, devido à sua eficácia, elevada durabilidade e baixo custo. A selecção dos geossintéticos a aplicar na cobertura e impermeabilização do corpo de resíduos tem sido feita com base nas suas propriedades físicas, nomeadamente na espessura no caso das geomembranas e da massa por unidade de área (m.u.a.) para os geotêxteis. Nas últimas décadas percebeu-se a necessidade da avaliação da durabilidade dos geossintéticos, nomeadamente a sua capacidade de resistência às acções mecânicas, tais como as danificações que ocorrem durante a instalação (DDI), pois estas podem comprometer o bom funcionamento da estrutura. Este estudo tem como objectivo aumentar o conhecimento existente sobre a DDI em geomembranas em polietileno de alta densidade (PEAD) e em geotêxteis de poliproprileno (PP) não tecidos. Procurou-se ainda entender a influência do tipo de solo e da m.u.a. do geotêxtil na danificação associada aos processos de instalação. Os ensaios de danificação laboratorial foram realizados de acordo com os procedimentos descritos na norma EN ISO 10722: 2007. Como forma de avaliar a DDI realizaram-se ensaios de caracterização mecânica a curto prazo, mais especificamente ensaios de tracção e de rasgamento. Este trabalho está dividido em três capítulos. No primeiro capítulo faz-se uma introdução geral aos geossintéticos, referindo os diversos tipos existentes e as suas funções. Procura-se ainda abordar os diversos elementos constituintes de um aterro de RSU. No segundo capítulo descrevem-se as propriedades características dos geossintéticos e continua-se a abordagem aos elementos constituintes de um aterro de RSU, realizando-se o seu dimensionamento. No terceiro capítulo realiza-se uma breve descrição dos procedimentos aplicados nos diferentes ensaios, faz-se uma análise experimental da influência da DDI nos geossintéticos e realiza-se uma comparação com outros resultados de estudos já publicados, obtidos em ensaios laboratoriais. Por fim, apresentam-se as conclusões obtidas do estudo e sugerem-se possíveis desenvolvimentos futuros. ABSTRACT: Nowadays, municipal waste, in the impossibility of being recycled, finds as final destination the municipal solid waste landfills (MSW). Such waste can contain substances which, if not properly isolated from the surrounding environment, may lead to its contamination. Over the past few years the geosynthetic resulted in a good bet in sealing landfills of MSW, due to its efficiency, high durability and low cost. The selection of the geosynthetics to apply in the cover and waterproofing of the body waste has been done on basis of their physical properties, including the thickness of the geomembranes and the mass per unit area (mua) of the geotextiles. Nowadays, the need to evaluate the durability of Geosynthetics is clear, particularly its capacity to resiste mechanical actions, such as damage occurring during the installation (DDI) because they may jeopardize the good functioning of the structure. This study aims to increase the existing knowledge on DDI of geomembranes from high-density polyethylene (HDPE) and nonwovens geotextiles in polypropylene (PP). It was also tried to understand the influence of soil type and the MUA of the geotextile in damage associated with the installation procedures. The laboratory tests of damage were conducted in accordance with the procedures described in EN ISO 10722: 2007. In order to assess the DDI induced, tests to characterise the short term mechanical behaviour of the geosynthetics were carried out, more specifically tensile tests and tearing tests. This work is divided into three chapters. In the first chapter a general introduction to geosynthetics is done, and its different types and its functions are briefly described. The elements that constitue a landfill of MSW are also referred. In the second chapter the main properties and characteristics of the geosynthetics are referred and the constituent elements of a landfill of MSW are described and some methods for its designed are included. The third chapter includes a brief description of the procedures of the various tests carried out in this work, an experimental analysis of the influence of DDI on the geosynthetics is done and the results obtained are compared with results from other studies published, obtained also in laboratory tests. Finally, the main conclusions of this work are presented and possible future developments are suggested.

Azijos žemyne formuojasi pasaulio geopolitinis-ekonominis centras. Vienos svarbiausių valstybių šiame procese yra Kinijos Liaudies Respublika (toliau Kinija), Indija ir Pietų Korėja. Geopolitinio-ekonominio centro formavimosi laikotarpiu komplikuotas ir probleminis tampa valstybių ekonominio saugumo užtikrinimas. Iškyla klausimas kokias priemones ir kuo remiantis pasitelkia šios valstybės savo ekonominio saugumo stiprinimui. Šio tyrimo objektas yra Kinijos, Indijos ir Pietų Korėjos ekonominė diplomatija. Darbo tikslas: pritaikyti ir kritiškai įvertinti Herscher-Ohlin-Vanek (HOV) modelį įvykdant Kinijos, Indijos ir Pietų Korėjos ekonominės diplomatijos lyginamąją analizę po įstojimo į Pasaulio Prekybos Organizaciją. Tikslui pasiekti iškeliami šie uždaviniai: 1) ištirti saugumo kaitą ir ekonominės diplomatijos sampratą; 2) įvertinti HOV modelį bei jo metodologinį pritaikymą Kinijos, Indijos ir Pietų Korėjos ekonominės diplomatijos analizei; 3) išanalizuoti esminius Kinijos, Indijos ir Pietų Korėjos ekonominės diplomatijos bruožus; 4) ištirti Kinijos, Indijos ir Pietų Korėjos ekonominės diplomatijos įrankių panaudojimą ekonominiam saugumui užtikrinti; 5) empiriškai palyginus Kinijos, Indijos ir Pietų Korėjos ekonominės diplomatijos įrankius kritiškai įvertinti HOV modelį. Tyrimui naudojami metodai: aprašomasis metodas, kokybinių ir kiekybinių metodų sintezė, ekonominės diplomatijos HOV modelis bei lyginamasis metodas. Atlikus tyrimą gauti rezultatai, jog HOV modelis daugeliu... [toliau žr. visą tekstą]
In Asia develops new world geopolitical-ecanomic centre. Of most important countries in this process are China, India and South Korea. Complicated period of geopolitical-economic centre formation rises vulnerabilies and challenges for the assurance of these countries economic security. Rises questions that means and why use these countries for their economic security strengthening. The object of this research is the economic diplomacy of China, India and South Korea. The goal is: to use and critically evaluate Herscher-Ohlin-Vanek model after accomplished comparative analysis of China, India and South Korea economic diplomacy after their join to the World Trade Organization. The tasks are: 1) to survey the fluctuation of security and conception of economic diplomacy; 2) to evaluate HOV model and its methodological application to the analysis of China, India and South Korea economic diplomacy; 3) to analyse essential features of China, India and South Korea economic diplomacy; 4) to examine China, India and South Korea economic diplomacy tools appliance for the assurance of economic security; 5) after empirical comparision of China, India and South Korea economic diplomacy tools to critically evaluate HOV model. For the research are used: descriptive method, the synthesis of qualitive and quantitive methods, economic diplomacy HOV model and comparative analysis. After research obtained results show that HOV model in most of cases can be used to explain countries‘ economic... [to full text]

This thesis is about social class and economic inequality, using the Goldthorpe class schema. It tests theories claiming that social class is increasingly irrelevant to inequality and people's life-chances with data on incomes and material living standards from the British Household Panel Survey. It covers the period over which the survey ran, i.e. 1991-2008. During this time many prominent social theories dismissed class analyses while others sought to retain the class concept but dismissed its economic foundations, seeking to ground it in culture instead. Economic inequality has not figured highly on the agenda of class analysts, at least not those working with the Goldthorpe class schema. There is a substantial body of work on mobility, voting behaviour, income poverty and material deprivation, but inequality in a broader sense has for the most part been neglected. This thesis is a step towards rectifying this situation. Thus it provides new information about within-career social mobility as well as income inequality within and between classes, on whether income mobility reduces class inequalities over time, and cast light on class inequalities in material living standards. The findings suggest that class is far from irrelevant to economic inequality. Class differences in incomes are persistent, between class inequalities contribute more to inequality overall than within-class inequalities, and while income mobility does reduce class inequalities over time it is not to the extent that supports the hypothesis that class is irrelevant to people's economic fortunes.

Heterozygous mutations in the glucocerebrosidase (GBA) gene represent the most common risk factor for Parkinson’s disease (PD), a disease in which midbrain dopaminergic neurons are preferentially vulnerable. However, the mechanisms underlying this association are still unknown, mostly due to the lack of an appropriate model of study. In this thesis, we aimed at elucidating the role of heterozygous GBA mutations in PD using a specific human induced pluripotent stem cell (hiPSC)-based model of disease. First we developed a protocol for the efficient differentiation of hiPSCs into dopaminergic cultures, and extensively characterized the derived dopaminergic neurons which expressed multiple midbrain relevant markers and produced dopamine. Next we screened a clinical cohort of PD patients to identify carriers of GBA mutations of interest. Using for the first time hiPSCs generated from PD patients heterozygous for a GBA mutation (together with idiopathic cases and control individuals) we were able to efficiently derive dopaminergic cultures and identify relevant disease mechanisms. Upon differentiation into dopaminergic neuronal cultures, we observed retention of mutant glucocerebrosidase (GCase) protein in the endoplasmic reticulum (ER) with no change in protein levels, leading to upregulation of ER stress machinery and resulting in increased autophagic demand. At the lysosomal level, we found a reduction of GCase activity in dopaminergic neuronal cultures, and the enlargement of the lysosomal compartment in identified dopaminergic neurons suggesting a decreased capacity for protein clearance. Together, these perturbations of cellular homeostasis resulted in increased release of α-synuclein and could likely represent critical early cellular phenotypes of Parkinson's disease and explain the high risk of heterozygous GBA mutations for PD.

Mestrado em Engenharia Civil
Já alguns anos percebeu-se a necessidade da avaliação da durabilidade dos geossintéticos, nomeadamente a sua capacidade em sobreviver aos danos durante a instalação (DDI). Como na maioria das vezes, os geossintéticos são instalados em contacto com materiais geotécnicos, a DDI é inevitável. Neste trabalho foi feita análise experimental através do ensaio de corte em plano inclinado, do desempenho de geossintéticos submetidos a DDI, caracterizando-se assim o efeito da DDI na capacidade de transferência de esforços entre o geossintético e o solo adjacente quando a rotura de corte ocorre num plano inclinado. Para tanto foram ensaiadas amostras de geossintéticos intactos e previamente danificados em campo. A campanha de ensaios incluiu a caracterização de 3 tipos de geossintéticos, sendo 2 geogrelhas e 1 geotêxtil. Os efeitos da DDI no ângulo de atrito máximo entre o solo e as amostras de geossintético são apresentados e discutidos. São explicitados os valores para coeficientes de redução para o ângulo de atrito na interface solo-geossintético, determinados a partir dos resultados dos ensaios. Os resultados do estudo levam a crer que a variação na capacidade de transferência de esforços em movimento de corte em plano inclinado entre os geossintéticos e o solo devido a DDI é baixo e bastante inferior ao que preveem os métodos de dimensionamento correntes.
The need to evaluate the geosynthetics durability, particularly their ability to survive damage during installation (DDI) has been realised as relevant for some years. As in most cases, geosynthetics are installed in contact with geotechnical materials, DDI is inevitable. In this work an experimental analysis of the performance of geosynthetics submitted to DDI using the inclined plane shear tests was done. This way it was possible to characterize the effect of DDI on the load transfer from the soil to the geosynthetic when the movement between them is in inclined plane shear. With this objective samples of geosynthetics both intact and damaged in the field, were tested. The test campaign included 3 geosynthetics, particularly 1 geotextile and geogrid. The effects of DDI on the maximum friction angle between soil and the geosynthetic are presented and analyzed. The reduction factor values for DDI determined from the tests results are presented as well as the main conclusions. The results from study suggest that the variation due to DDI in soil-geosynthetics interface strength is small and likely to be smaller than the expected when current design methods predict.

The thesis is in two main sections. In the first section, studies on methylarginine processing enzymes are presented. Dimethyalrginine dimethylaminohydrolase (DDAH) is a class of enzymes involved in the metabolism of methylarginines ADMA and L-NMMA, which indirectly regulate physiological nitric oxide levels. It is desirable to inhibit excess NO in pathological situations, and the arginine mimetic L-257 is a DDAH inhibitor which reduces levels of NO. Synthesis of ester analogues of L-257 proved to be troublesome with a low yielding key guanidine forming reaction. However, amide analogues were readily synthesised, and testing for DDAH inhibition showed the dimethylamide analogue possessed similar activity to L-257. Further design and synthesis of a 7-membered cyclic analogue, based on the crystal structure of huDDAH1 with L-257, provided a novel analogue with no significant inhibition for rat kidney DDAH. Purified and isolated huDDAH2 protein showed activity after incubation with substrate L-NMMA. In the second part studies on aldehyde auto-oxidation are presented. Aldehydes autoxidise to their acids, via an acyl radical, which can undergo addition reactions with electron-deficient acceptors in a radical hydroacylation reaction. An α- iodo and α-chloro hexanal failed to autoxidise, however 7-hydroxycitronellal readily autoxidised and added to pentafluorophenyl(PFP)-vinyl sulfonate. Further studies on hydroacylation of butanal with PFP-vinyl sulfonate led to functionalised β-ketosulfonates which undergo elimination to form an enone and can then undergo further conjugate addition in situ by nucleophiles. Conjugate addition was carried out using carbon, nitrogen, oxygen and phosphorus nucleophiles, providing a method of obtaining products which are challenging to make via hydroacylation of electron-rich alkenes. Decarbonylation of pivaldehyde to the t-butyl radical, via auto-oxidation, was optimised and the alkyl radical captured by a number of electron-deficient acceptors, providing a complementary method to current methods of t-butyl addition using metal reagents.

Nesta tese foi estudado o espalhamento Raman intensificado pela superfície (SERS) em regime de detecção de uma molécula em eletrodo de prata ativado por ciclos de oxidação e redução. Neste regime, de baixas concentrações, são observadas intensas flutuações de intensidade SERS as quais foram controladas neste substrato pela aplicação de potencial ao eletrodo, o que foi associado a alterações na concentração de moléculas adsorvidas na superfície do eletrodo. Além da dependência com o potencial aplicado, foram estudadas através de simulações Monte Carlo, a contribuição nestas flutuações da constante de adsorção das moléculas, do número de \"hot spots\" (regiões de altas intensificações SERS) e do tipo de \"hot spot\" (em termos de eficiência para detecção de espectros de uma molécula). Através destas simulações foram verificadas flutuações de intensidade muito semelhantes às observadas experimentalmente. Além das flutuações de intensidade foram também observadas flutuações de intensidades relativas, como por exemplo, das relações de intensidades anti-Stokes/Stokes, as quais foram interpretadas segundo um modelo de ressonância, através do qual foi possível estimar as energias de ressonância nos \"hot spots\". Alguns dos resultados indicaram a contribuição de ressonâncias finas, as quais foram interpretadas como resultado de interferências entre ressonâncias de plasmon de superfície. Interferências como estas foram demonstradas através de simulações pelo método DDA (\"Discrete Dipole appoximation\") em modelos simples de \"hot spots\" formados por nanobastões de Au.
In this thesis it was studied surface-enhanced Raman scattering (SERS) at single-molecule detection on Ag electrode activated by oxidation and reduction cycles. At this low concentration limit it was observed strong SERS intensity fluctuations that were controlled by the applied potential to the electrode and this control was associated to changes in surface concentration of adsorbed molecules. Furthermore, it was studied through Monte Carlo simulations the influence of adsorption constant, number of \"hot spots\" (regions of high SERS enhancements) and type of \"hot pot\" (in terms of efficiency for single-molecule detection). With such simulations, it was verified fluctuations of SERS intensities very similar to experimental observations. Besides absolute intensity fluctuations, we also observed fluctuations of relative intensities as, for instance, the. anti-Stokes to Stokes intensity ratios. These fluctuations were interpreted according to a resonance model, which made possible the estimative of resonance energies at the SERS \"hot spots\". Some of these results indicated the existence of sharp resonances that were interpreted as a result of interferences among surface plasmon resonances, which were demonstrated through DDA (Discrete Dipole Approximation) simulations in simple models of \"hot spots\" formed by Au nanorods

Ce mémoire porte sur la conception d'un accouplement à roue-libre. Il est destiné à remplacer l'embrayage piloté d'un véhicule hybride parallèle " ZEV ", simple arbre, à boîte de vitesses pilotée. Le premier chapitre définit le véhicule hybride étudié. Il développe ses modes de fonctionnement et montre la nécessité d'isoler le moteur thermique du reste de la chaîne de traction, en fonction des conditions de roulage. Le chapitre deux présente et compare deux solutions possibles : l'embrayage à sec piloté et la roue-libre. La mise au point de la solution à roue-libre a nécessité d'étudier les vibrations de torsion dans la transmission. Un outil modulaire spécifique a été développé sous MATLAB SIMULINK. Il est présenté dans le chapitre trois. Enfin, le chapitre quatre montre comment cet outil a été employé en phase de conception et présente une solution constructive. Cet accouplement doit être implanté dans un véhicule hybride prototype réalisé conjointement par le LAMIH et PSA
This thesis deals with the design of a free-wheel clutch. This unit is intended to replace the automated dry single-plate clutch of a parallel hybrid car with thermal and electric powertrain. Furthermore, the car is a single shaft zero emission vehicle fitted with a controlled gearbox. Chapter one focuses on the type of hybrid vehicle studied. It shows the need to isolate the engine from the rest of the drive train, depending on the driving conditions. Chapter two presents and compares the two alternatives : automated clutch and free-wheel. In order to develop the free-wheel option, the torsional vibrations in the automotive drive line had to be closely studied. It required the design of a specific modular tool, as presented in chapter three, with the help of MATLAB SIMULINK. Lastly, chapter four shows how this tool was used during the design stage and specifies the way to build it. The free-wheel is then to be fitted to a prototype hybrid vehicle, constructed by both the LAMIH and PSA

La spectroscopie par fluage thermostimule a permis l'etude des deux principaux modes de retard de composites modeles bille de verre/matrice polyepoxy dans le domaine de temperature 180c/+150c. La microstructure des composites a ete comparee a celle du reseau utilise comme matrice. L'influence de la presence du renfort et de son traitement de surface a ainsi pu etre isolee et discutee en termes d'interphase et de mobilite moleculaire. Dans le domaine vitreux, une etude de la structure fine du spectre de modes de retard a montre le caractere complexe du mode basse temperature et l'existence d'une zone d'influence autour du renfort a ete mise en evidence. L'etude du mode haute temperature a permis d'estimer l'epaisseur de cette interphase. La resolution experimentale de ce mode a montre des phenomenes caracteristiques de l'interface/interphase. Le nombre de configurations possibles et la longueur des sequences mobiles ont pu etre compares a ceux de la matrice polyepoxy sans renfort. La technique du fluage thermostimule se revele donc particulierement bien adaptee a l'etude fine de la microstructure des composites a matrice organique

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro Tecnológico. Programa de Pós-Graduação em Engenharia de Produção.
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As dificuldades na aprendizagem têm sido foco de recentes pesquisas. Uma das causas é o DDA (Distúrbio do Déficit de Atenção). A utilização da tecnologia, em que se destaca o uso do computador, também vem sendo alvo de discussões no processo de aprendizagem. A Psicologia enquanto ciência e profissão e com uma visão ampla da cognição, ou seja o ato de conhecer, integrar, elaborar e exprimir a informação, faz parte desse processo. Procurando discutir as questões acima, este trabalho analisa, em termos de recursos, como o computador pode contribuir para melhorar o setting terapêutico nas sessões de reeducação de crianças com DDA, com a intenção de fortalecer ainda mais o diálogo entre clínica - teoria - pesquisa, o que é de fundamental importância para o crescimento dessa área. Busca o conhecimento da teoria e da ética, através do estudo de caso; apresenta um modelo de trabalho no computador e verifica suas contribuições. Observando que o setting terapêutico não é um atributo inerente do computador, mas uma conseqüência que está vinculada com a forma com que ele é utilizado.

In order to resolve the Al void formation originated from the severe stress issues in dynamic random access memory (DRAM), double-deposited-aluminum (DDA) layer process was proposed. This novel metallization process can be effectively and simply performed with the native oxide such as Al 2 O 3 between upper and lower Al metal layer by ex-situ deposition technique. We could effectively control the Al void by adapting the DDA layers with different grain structure. From this novel metallization process, we have confirmed the optimal thickness of Al barrier metal to 100Å to be free from Al voids, which makes it possible to improve the static refresh characteristics of DRAM by 17%.

Les douleurs lombaires touchent 80% de la population au moins une fois dans la vie et constituent un véritable problème de santé publique pour nos sociétés modernes industrialisées. Ces douleurs lombaires sont, dans la grande majorité des cas, la conséquence clinique d'une atteinte des articulations vertébrales marquée par une dégénérescence progressive des disques intervertébraux. La connaissance des mécanismes menant à cette dégénérescence discale sont aujourd’hui de mieux en mieux connus et laisse envisager une prise en charge étiologique, et non plus uniquement symptomatique. Les premiers résultats concluants en ingénierie tissulaire du cartilage articulaire associés à l'existence de points communs entre le cartilage articulaire et le disque intervertébral laissent suggérer la possibilité de régénérer ce dernier, et en particulier le Nucleus pulposus (élément central du disque intervertébral), par ce même type d'approche. Le principe de cette approche est basé sur l'utilisation de cellules associées à un biomatériau. L'ensemble est injecté au sein d'un disque dégénéré afin de le restaurer. Un état des avancées actuelles dans ce domaine est réalisé et les différentes problématiques rencontrées au cours de la mise en œuvre de ce type de projet sont abordées. Il s'agit notamment du choix des cellules à injecter et des matrices associées, des conditions de culture adéquates, ainsi que du choix des méthodes d'évaluation et des modèles animaux
Low back pain affects 80% of the population at least once during life and constitutes a public health problem for our modern industrialized societies. Usually, they are the consequences of the intervertebral disc degeneration. Currently, the knowledges about mechanisms leading to this disc degeneration are well understood and allow to define new targets to treat the origin of the intervertebral disc degeneration. The first promising results in tissue engineering of articular cartilage associated with the existence of similarities between articular cartilage and intervertebral disc allow to considered the same approach to treat the intervertebral disc. The principle of this approach is based on the use of cells associated with a biomaterial and the substitute is injected into the degenerated disc. An update of current advances in this area is achieved and the various problems encountered during the development of such projects are discussed. These include the choice of cells and scaffolds injected, the choice of appropriate culture conditions, and the choice of evaluation methods and reliable animal models

Nitric Oxide, NO, is an important neurotransmitter and signaling molecule in almost every system of the human body. Its regulation is equally important as a dysfunction in regulation leads to pathophysiologic conditions and disease states. There are many ways that NO is regulated, but one of the more common methods is to regulate the enzyme that creates it, Nitric Oxide Synthases or NOS. Again, many ways to regulate this enzyme exist, but this work focuses on inhibition by an endogenous molecule produced via the normal process of protein degradation, Asymmetric Dimethyl Arginine or ADMA. Another enzyme in the body is responsible for metabolism of ADMA called Dimethylarginine Dimethylaminohydrolase or DDAH. By regulating the activity of DDAH, we can control concentrations of ADMA and therefore the inhibition of NOS, which in turn regulates the production of NO. It is the purpose of this project to create a probe that can be used in vivo for the DDAH enzyme family using organic synthesis to produce a product that, when effectively bound to the enzyme target, can also undergo a click chemistry reaction to tailor the utility of the probe itself. The Dixon Lab has designed such a molecule and the synthesis is what follows.

Embora a imunização gênica apresente vantagens sobre as vacinas convencionais, nem sempre a produção de anticorpos e a conseqüente proteção conferida pelo sistema imune contra o antígeno de interesse são satisfatórias. As causas da baixa eficácia da imunização gênica, nestes casos, ainda são pouco conhecidas, porém um dos pontos mais relevantes em todo o processo é a eficiência da transfecção do DNA. O sucesso desta metodologia depende do estabelecimento de um sistema de entrega de gene eficiente e seguro. De acordo com a teoria proposta de Kuhn et al. (1999), baixas concentrações de moléculas anfifílicas podem ser usadas para neutralizar a carga do DNA, sem a formação de lipossomos. Este trabalho teve como objetivo avaliar o uso do lipídio catiônico dimetildioctadecil brometo de amônio (DDAB) como neutralizador da carga do DNA para a imunização de camundongos. Para a realização dos experimentos, um plasmídeo expressando a proteína do capsídeo (CA) do vírus da artrite encefalite caprina (CAEV), sob o comando do promotor citomegalovírus humano (CMV), foi utilizado nas imunizações. Os camundongos foram divididos em cinco grupos e receberam, via subcutânea, três inoculações de 10 μg de DNA linear (grupo 1), DNA linear mínimo (grupo 2), DNA circular (grupo 3) associados a 2,7 μg de DDAB ou DNA circular nu (grupo 4). O grupo controle foi inoculado com 2,7 μg de DDAB em solução fisiológica. Os camundongos imunizados com o plasmídeo na forma linear associado ao DDAB apresentaram maiores títulos de anticorpos CA que os inoculados com a forma circular ou linear mínima. Os dados sugerem que o lipídio catiônico DDAB tem potencial de uso como veículo de transfecção gênica em animais.
Genetic immunization has several advantages over conventional vaccines. However, antibody production and protection against the antigen of interest are often not satisfactory. In these cases, the causes of the low effectiveness of the genetic immunization are still poorly understood. One of the most relevant points in the immunization process is the DNA delivery. A successful DNA vaccine depends on the establishment of an efficient and safe gene delivery system. In accordance with Kuhn et al. (1999), low concentrations of amphiphilic molecules can be used to neutralize the DNA charge without the formation of liposomes. The aim of this study was to examine the effect of cationic lipid dimethildioctadecyl-ammonium bromide (DDAB) as a charge neutralizer for DNA for the immunization in mice. A plasmid expressing the capsid protein (CA) of caprine arthritis encephalitis virus (CAEV) under the control of the human cytomegalovirus immediate early promoter (hCMVie1) was used in the immunization. Mice were grouped and immunized with three injections of either 10 μg of linear DNA (group 1), minimal linear DNA (group 2) or circular DNA (group 3) associated with 2,7 μg of DDAB or naked circular DNA (group 4). The control group received 2,7 μg of DDAB diluted in physiologic solution. Mice immunized with DNA in linear form associated with DDAB developed higher titers of antibodies against CA than the animals immunized with the circular or minimal linear form. The data suggest that lipidic cationic DDAB could be a useful delivery system for genetic immunization.

Pseudomonas aeruginosa (PA) est la troisième bactérie impliquée dans les maladies nosocomiales et est la principale cause de mortalité des patients atteints de la mucoviscidose. PA est résistante à la plupart des traitements antibiotiques. Trouver de nouvelles stratégies thérapeutiques est devenu un enjeu majeur de santé publique, l’une d’entre elles est l’inhibition de facteurs de virulence. Parmi ceux-ci, les lectines sont des protéines impliquées dans l’adhésion et la formation de biofilm via des interactions avec des sucres (PA-IL, PA- IIL, FliC, FliD, PilA, PilY1 et CupB6).Le but de ce travail est donc de trouver des leurres moléculaires ayant une forte affinité pour ces lectines. Ceux-ci sont des motifs saccharidiques présentés de façon multivalente : glycoclusters. De part leur grande diversité structurale et leur faible quantité, un outil de criblage innovant a été développé qui consiste en une lame de verre microstructurée : le glycocluster-microarray. Les glycoclusters sont immobilisés de manière ordonnée par DNA Directed Immobilization (DDI). Deux méthodes de criblage ont été développées grâce à cet outils : 1) le criblage en solution et par compétition d’une bibliothèque de motifs saccharidiques et 2) le criblage d’une bibliothèque de glycoclusters immobilisés sur le microarray. Avec cet outil, des protocoles de mesures d’IC50 et de Kd ont aussi été fiabilisés pour caractériser les meilleurs candidats inhibiteurs des lectines. Le glycocluster- microarray présente l’avantage de n’utiliser qu’une très faible quantité de matériel (quelques picomoles) et permet de réaliser diverses analyses en parallèle.Afin de valider cet outil, une étude sur l’impact de la densité de surface en glycocluster a été menée. Le criblage de plus de 150 motifs saccharidiques a permis de sélectionner ceux ayant une forte affinité pour les lectines. L’analyse sur microarray complétée par de la modélisation moléculaire d’une bibliothèque de glycoclusters, possédant ces motifs et différentes topologies, valences et propriétés (aromaticité, charge,…), a permis d’identifier les paramètres clés dirigeant les relations structure-affinité. Une activité anti-biofilm chez PA a été démontrée avec les meilleurs glycoclusters ciblant PA-IL.Tester l’activité in vivo, chez l’animal, des meilleurs candidats est une voie à explorer. Cibler d’autres lectines comme celles présentes sur le flagelle et les pili de PA et notamment impliquées dans son adhésion précoce est aussi une voie à développer. Pour cela, des tests préliminaires ont été présentés et d’autres sont en cours faisant appel à l’utilisation de bactéries entières ainsi qu’à une détection sans marquage des lectines
Summary: Pseudomonas aeruginosa (PA) is the third pathogen involved in nosocomial diseases and the major cause of mortality of cystic fibrosis patients. PA develops resistance to antibiotics treatments. And so, developing new therapeutic strategies is a public health issue. One of the promising strategies is to inhibit virulence factors involved in the adhesion and the biofilm formation of PA. Some of these virulence factors are lectins which interact with sugars (PA-IL, PA-IIL, FliC, FliD, PilA, PilY1 and CupB6).The goal of this work is to find molecular decoys which have a strong affinity for these lectins. These are saccharidic units with a multivalent display: glycoclusters. An innovative screening tool has been developed: the glycocluster-microarray, to study lectin/glycocluster interactions. It is a microstructured glass slide where glycoclusters are immobilized by DNA Directed Immobilization (DDI). Two screening methods have been developed with this microarray: 1) the screening in solution and by competition of a saccharidic units library and2) the screening of a glycoclusters library immobilized on the microarray. Protocols of IC50 and Kd measurements have also been developed with this tool to characterize the best lectins inhibitors. This tool allows to use few amount of material (few picomoles) and to do parallel analysis.To validate the microarray, a study of the impact of glycoclusters surface density has been done. The screening of more than 150 saccharidic units allowed the selection of the ones that display the best affinity forlectins. The analysis, on microarray and molecular simulations, of the glycoclusters library displaying thesesaccharidic units and several topologies, valences and properties (aromaticity, charge,…) enable to identify key parameters of structure-affinity relationships. An anti-biofilm activity has been observed for the best glycoclusters targeting PA-IL.Testing in vivo activity of these best candidates will be explored. Targeting others lectins such as the ones on the flagella and pili of PA and involved in the early adhesion needs also to be developed. To this end, preliminary tests have been showed and some are in progress

The Western Tropical North Atlantic (WTNA) was once thought to be a net source of carbon dioxide (CO₂) to the atmosphere, but recent studies have shown that this Amazon River influenced region may actually act as a net sink for CO₂. During a 2010 research cruise to the WTNA, we characterized the impact of the Amazon River on offshore diazotrophy (N₂-fixation) and the resulting stimulation of biological carbon export from surface waters. Through the delivery of phosphate- and silicate-replete waters to the nitrogen (N) limited surface waters of the WTNA, the aging Amazon River plume promotes the growth of diatom-diazotoph associations (DDAs). Regions supporting large DDA blooms were associated with increased pCO₂ and DIC drawdown in the surface waters, reflecting the net export of carbon from the mixed layer. The existence of this biologically mediated linkage between the C and N cycles in productive surface waters is well known, but we have only recently discovered a stimulatory relationship in deep waters between oil/gas release and N₂-fixation. This association was first observed after the Deepwater Horizon oil spill in 2010 and we again saw evidence for it in the days following the Hercules 265 natural gas blowout. This blowout event was characterized by the release of an unknown quantity of natural gas into the shelf waters of the Northern Gulf of Mexico, but we detected a response from the marine microbial community within days. We observed a significant drawdown of dissolved oxygen and found biogeochemical evidence for the incorporation of methane-carbon into the food web, along with a modest stimulation of N₂-fixation. The episodic nature of anthropogenic blowouts makes them difficult to study, so we use cold seeps in the Gulf of Mexico as natural analogues. Interestingly, we have measured both methane oxidation and N₂-fixation at depth above some of the more active seeps. Using NanoSIMS analyses, we have taken the first steps towards physically characterizing the organisms utilizing these metabolisms. It appears that different organisms are carrying out these processes, with CH₄-assimilation occurring primarily in individual particles or small aggregates, whereas N₂-fixtion was associated with larger, sulfur-containing aggregates. Continued NanoSIMS work in combination with the use of microbial ID techniques will help to further characterize these unique deepwater diazotrophs.

Procedural Content Generation (PCG) and Dynamic Difficulty Adjustment (DDA) have been used separately in games to improve player experience. We explore using PCG and DDA together in a feedback loop to keep a player in the "flow zone." The central tenet of this work is a conjecture about how the shape of the performance versus difficulty curve changes at the boundaries of the flow zone. Based on this conjecture, we have developed an algorithm that detects when the player has left the flow zone and appropriately adjusts the difficulty to bring the gameplay back into flow, even as the skill of the player is changing. We developed a game-independent algorithm, implemented our algorithm for the open-source Infinite Mario Bros (IMB) game and conducted a user study that supports the hypothesis that players will enjoy the game more with DDA - PCG algorithm.

A síntese de imagens é uma ferramenta valiosa na compreensão de diversos fenômenos da natureza. Nos últimos anos várias abordagens têm sido propostas para sintetizar tais fenômenos. A grande maioria de tais abordagens têm se centralizado no desenvolvimento de modelos procedurais. Porém, cada uma destas técnicas simula exclusivamente um fenômeno natural. Um dos métodos de síntese de imagens fotorealísticas mais proeminente é denominado de Traçado de Raios (Ray Tracing). Contudo, apesar de produzir imagens de excelente qualidade, este método é computacionalmente muito oneroso. A síntese de fenômenos naturais utilizando-se o traçado de raios é um desafio. É importante que este problema seja abordado, apesar da sua complexidade, pois a simulaçao fotorealista da natureza é muito importante para os cientistas e pesquisadores desde o surgimento dos computadores. Um algoritmo versátil e rápido para a síntese de fenômenos da natureza através do traçado de raios utilizando campos de altitude é proposto. O algoritmo utiliza uma modificação do algoritmo do Analisador Diferencial Digital de Bresenham para atravesar uma matriz bidimensional de valores de altitude. A determinação das primitivas geométricas a serem interseccionadas por um raio é obtida num tempo ( N ) , sendo N o número de altitudes no campo de altitude. Este trabalho faz uma comparação em termos de velocidade e realismo deste método com outras abordagens convencionais; e discute as implicações que a implementação deste método traz. Finalmente, destaca-se a simplicidade e versatilidade que este método proporciona devido à pequena quantidade de parâmetros necessária para a síntese de fenômenos naturais utilizando o traçado de raios. Para a criação de animações basta a especificação de novos parâmetros num intervalo de tempo diferente.
Visualization is a powerful tool for better undestanding of several natural phenomena. In recent years, several techniques have been proposed. Considerable interest in natural scene synthesis has focused on procedural models. However, these techniques produce synthetic scenes of only one natural phenomenon. Ray tracing is one of the most photorealistic methods of image syntesis. While providing images of excellent quality, ray tracing is a computationally intensive task. Natural scene synthesis is a challenging problem within the realm of ray tracing. It is important to tackle this problem, despite of its complexity, because photorealistic simulation have been important to scientific community since the appearance of computers. A fast and versatile algorithm for ray tracing natural scenes through height fields is presented. The algorithm employs a modified Bresenham DDA to traverse a two dimensional array of values. The objects tested for intersection are located in ( N ) time where N is the number of values in the field. This work compares the speed-up and photorealism achieved in natural scene synthesis using this method with other algorithms and discusses the implications of implementing this approach. As a final point, the simplicity and versatility of synthesizing complex natural scenes from a few parameters and data is especially attractive. Animated sequences require only the additional specifications of time modified parameters or data.

Nitric Oxide is an important molecule in human cells. It is responsible for multiple functions in various systems including the cardiovascular, immune, nervous, digestive, and reproductive systems. Nitric oxide is synthesized by the enzyme nitric oxide synthase, and its activity is controlled by the levels of asymmetric dimethyl arginine (ADMA) which in turn are controlled by Dimethylaminohydrolase (DDAH). DDAH is an important enzyme for study due to its ability to indirectly control nitric oxide synthase. 2-chloroacetimidine is an inhibitor of DDAH. Synthesizing molecules that contain a 2-chloroacetimidine moiety and a retrievable chemical tail could prove to be instrumental in further studying DDAH and other enzymes with similar reactivity in their active sites.

碩士
國立陽明大學
生物化學研究所
86
p53為一抑癌基因，當細胞DNA損害時p53常會大量表現導致G1 arrest或apoptosis。由本實驗室所建立的可持續表現受溫度調控的p53 mutant的轉型細胞株1-5，利用而tNA差異展現法(differential display)我們選殖到可受p53調控的基因mDDA3。經DNA序列分析，發現血DDA3的cDNA全長為1.8kb。再由human EST database找到了一個EST clone將其定序並與老鼠的DDA3比對後，發現二者之胺基酸序列具有67.9%的相似性，並且皆含3個SH3 binding motif PXXP。Colony的rmation assay結果顯示DDA3大量表現可以抑制H1299人類肺癌細胞的生長。將mDDA3的N端接上FLAGpeptides，以immunofluorescence觀察可見而JDA3在細胞質中表現。在invitro及in vivo下做轉錄和轉譯，可得大小約39kD和43kD的蛋白，這和由DDA3的open reading frame所預測出的蛋白質大小接近。利用human及mouse DDA3的相似區域我們設計了二條peptides，將其與BSA結合後注射兔子可以測得血清中有抗體。至於其與DDA3蛋白之結合，則有待更進一步的實驗確認。 p53 tumor suppressor is a transcription factor that functions through activation of a number of downstream target genes. We have previously identified and cloned a p53 regulated gene, DDA3, through differential display of the mRNAs of the IW32 mouse erythroleukemia cells grown in the presence or absence of wild type p53. The mouse DDA3 cDNA (mDDA3) is about 1.8 kb in size. Two alternatively spliced transcripts were cloned, mDDA3s contains an open reading frame of 329 amino acids and mDDA3L has an additional ATG at 45 bp upstream of the translation initiation codon of mDDA3s. Through homology search of the human EST database, an EST clone was identified and sequenced. This cDNA showed 69% identity in protein sequence with mDDA3, and may represent the human homologue of mDDA3. In vitro transcription and translation showed that mDDA3 and hDDA3 could both encode proteins of 39kDa in size. When mDDA3 was fused to the Flag tag at its 5' terminus and introduced into cells, a protein of 43kDa that reacted with anti-Flag antibody was found. Immunofillorescence indicates that DDA3 was distributed in the cytosol. Over-expression of DDA3 suppressed the growth of H1299 human non-sinall-cell lung cancer cells, as analyzed by colony formation assay. These results suggest that DDA3 may mediate, at least in part, the growth suppressing function of p53.

碩士
國立陽明大學
生物化學研究所
87
Wild type p53 expressed from a temperature-sensitive (tsp53) construct induces both G1 cell cycle arrest and apoptosis in the p53-negative IW32 mouse erythroleukemia cell line. Using PCR-based differential display analysis, we previously identified a new p53-inducible gene, DDA1, whose mRNA was upregulated in tsp53-transfected IW32 cells following induction of wild type p53 expression by temperature shift to 32°C. The DDA1 mRNA induction was detectable within 1 hour after temperature downshift, and rapid degradation was observed when the temperature was shifted back to 37°C, suggesting that the expression of DDA1 is dependent on the continuous presence of p53. The DDA1 mRNA was also induced in DNA damaging reagent-treated NIH3T3 cells. Previous studies from this lab have shown that mouse DDA1 cDNA predicted to encode a protein of 498 amino acid residues containing 12 transmembrane domains. The loop region between 6th and 7th transmembrane domains was used as immunogen to produce rabbit polyclonal antibodies, and antibodies that can recognize E. coli expressed mDDA1 protein was obtained. Immunofluorescence analysis indicated that DDA1 protein is located in the cytoplasm. Overexpression of mDDA1 cDNA in H1299 cells inhibited cell growth，as shown by the colony formation assay. The cDNA of human DDA1 that is 73 % identical to mouse DDA1 was acquired by library screening and database searching. The two amino acid sequences share 90 % identity.

碩士
國立陽明大學
生物化學研究所
88
Abstract p53 tumor suppressor is a transcription factor that causes cell growth arrest and induces apoptosis. Identification of the p53 downstream target genes is therefore important to unravel the mechanisms underlying p53 actions. We have previously identified and cloned a p53-regulated gene, DDA1, by the RNA differential display of an IW32 erythroleukemia stable clone (1-5) that contains a temperature-sensitive p53 mutant gene, tsp53val135. Sequence comparison revealed that mDDA1 shares 73% and 90% identity in its nucleotide and protein sequences, respectively, to the newly identified human thiamine transporter gene (hTHTR-1). To further investigate the subcellular localization and function of DDA1, we established DDA1 expressing clones under the control of the tetracycline inducible promoter. After 24 hours treatment of 2 mg /ml Doxycycline, clone 118 transfectant cells could be induced to express DDA1 mRNA and protein. Immunofluorescence analysis indicated that DDA1 was present on the plasma membrane. Growth of DDA1 stable transfectant was partly inhibited in the presence of Doxycycline. Ability of the clone 118 cells to uptake thiamine increased 2-fold in the presence of Doxycycline. These data demonstrated that mDDA1 possesses thiamine transporter activity. hTHTR-1 mRNA was induced by DNA damage in a p53 dependent manner. Induction was detected in 293 cells expressing endogenous p53, but not in 293T cells whose p53 was inactivated. Together these results indicate that the high affinity thiamine transpoter is a p53 regulated gene. 中文摘要 ..................................................... 1 英文摘要 ..................................................... 2 緒論 ..................................................... 3 實驗材料 ..................................................... 16 實驗方法 ..................................................... 18 實驗結果 ..................................................... 30 實驗討論 ..................................................... 36 參考文獻 ..................................................... 40 附圖 ..................................................... 47

碩士
國立陽明大學
生化暨分子生物研究所
94
The aim of this study is to investigate the effects of DDA3 on microtubule (MT) dynamics and functions. DDA3 is a p53 transcriptional target shown to be colocalized with MT in cells and bind MT plus end binding protein EB3. By immunofluorescence analysis and GST pull-down assay, we have mapped the MT and EB3 binding domains of DDA3. Both of them are amino acids 118-329. Ectopic expression of DDA3 in cells induced nocodazole-resistant MT bundles characterized by acetylation; in vitro MT bundling assay followed by electron microscopy analysis demonstrated that DDA3 directly bundles and stabilizes MT. DDA3 is expressed in the brain in a p53-independent manner. To examine if DDA3 is involved in MT-mediated activities, we explored the role of DDA3 in neurite formation using PC12 pheochromocytoma and N2a neuroblastoma cells. Overexpression of DDA3 inhibited neurite formation during NGF-induced PC12 differentiation, in contrast, knockdown of DDA3 expression by small interference RNA resulted in neurite outgrowth in N2a cells. Consistent with these findings, DDA3 expression was down-regulated during retinoic acid induced N2a differentiation. Interestingly, blocking the expression of EB3 suppressed the neurite outgrowth during induced differentiation of N2a cells. Together this study uncovers a novel role of DDA3 in MT dynamics and MT-mediated functions such as neurite formation.

碩士
中原大學
土木工程研究所
86
Many engineering structures near the mountain region of Taiwanhave been subjected to the threat of rock-fall. In order to economically and efficiently install protection means, identificationof rock-fall source area as well as prediction of falling rock pathsis essential. In this thesis, the ROCKPATH program was written to analyze the behavior of a single falling rock, and the DDAD code was adopted to study the effects of multiple falling rocks. Taiwan is located between Euro-Asia and Philippine plates, and two-thirds of its area is covered by mountains. Because of local geological and climate conditions, landslides of different types occurs frequently, and among them, rock-fall is more typical. In literature, many rock-fall simulation programs were developed by foreign scholars and were merely capable of handling one single falling rock, neglecting the effects of multiple falling rocks. Based on the lumped-mass method for a single falling rock, ROCKPATH was developed, and its validity was verified by several illustrative examples. As a special version of the Discontinuous Deformation Analysis for Disks, DDAD can analyze the complex interactions of thousands of particles. The task of modifying the velocity components of a particle after impacting a slope enabled DDAD to simulate rock-fall problems, and DDAD simulation results for single falling rock examples were well comparable to those computed by ROCKPATH. In the additional three examples provided in this thesis, DDAD computation results revealed that the effects of multiple falling rocks could not be ignored and the protection design based on single falling rock mode would not be conservative. Moreover, the normal restitution coefficient was back-calculated by DDAD for a falling rock case in Taiwan.

博士
國立陽明大學
生化暨分子生物研究所
95
We have previously identified mouse DDA3 as a p53-inducible gene. To explore the functional role of DDA3, we screened a mouse brain cDNA library by the yeast two-hybrid assay, and identified the microtubule plus-end binding protein EB3 as a DDA3 interacting protein. Binding of DDA3 to EB3 was verified by GST pull-down assay and subcellular co-localization; co-immunoprecipitation further indicated that interaction of these two proteins within cells required intact microtubules. Domains of DDA3-EB3 interaction were mapped by GST pull-down assay to amino acids (a.a.) 118-241 and 242-329 of DDA3 and the N- and C-termini of EB3. Immunofluorescence analysis revealed co-localization of DDA3 with microtubules in various cell phases, and regions encompassing a.a. 118-241 and 242-329 contained microtubule-interacting and bundling activities. In vitro microtubule binding assay showed that DDA3 and EB3 associated directly with microtubules, and cooperated with each other for microtubule binding. In addition, DDA3 bound to the EB3 interacting partner APC2, a homologue of the tumor suppressor adenomatous polyposis coli (APC) which is a component of the β-catenin destruction complex. Ectopic expression of DDA3 and EB3 enhanced β-catenin-dependent transactivation and cyclin D1 production, while knockdown of endogenous DDA3 or EB3 inhibited β-catenin-mediated transactivation and the ability of cells to form colonies, implicating DDA3 and EB3 in the β-catenin-mediated growth signaling. Furthermore, overexpression of DDA3 inhibited neurite formation during NGF-induced PC12 differentiation, whereas knockdown of DDA3 expression by small interference RNA resulted in neurite outgrowth in N2a cells. Consistent with these findings, DDA3 expression was downregulated during retinoic acid induced differentiation of N2a neuroblastoma cells. Together, our results uncover novel roles of DDA3 in microtubule dynamics, cell growth and neuronal differentiation.

碩士
國立陽明大學
生化暨分子生物研究所
96
Abstract mDDA3 is a p53 transcriptional target that interacts with ASPP2. Using GST pull-down assay, we now mapped the interaction domains to amino acids 118-241 of DDA3 and the ASPP2 C-terminal region (aa 900-1134) shown to interact with p53. Transfection reporter assay using the Bax promoter-driven luciferase construct indicated that the ASPP2-stimulated luciferase activity was inhibited by the expression of mDDA3. In vitro binding assay showed that mDDA3 inhibited the stimulation of ASPP2 on p53 transactivity without affecting its binding to p53. mDDA3 is a microtubule-associated protein that stabilizes microtubules and is expressed in the brain in a p53-independent manner. Previous finding from this lab indicates that overexpression of mDDA3 inhibited neurite formation during NGF-induced PC12 cell differentiation, in contrast, knockdown of mDDA3 expression by small interference RNA resulted in increased neurite outgrowth in N2a cell. To examine if mDDA3 is involved in neuritogenesis, we explored the role of mDDA3 in neurite outgrowth and axon formation using cultured rat hippocampus neurone. Overexpression of mDDA3 significantly inhibited neurite outgrowth and axon formation, in contrast, knockdown of mDDA3 promoted neurite outgrowth and axon formation in hippocampus cells. Similarly, treatment with the microtubule-stabilizing drug taxol led to microtubule bundling and suppression of neurite outgrowth in hippocampus cells. Together these finding show that mDDA3 plays an important role in neurite outgrowth and axon formation.