Tesis por compendio / [ES] Desde el origen de los cítricos, hibridaciones naturales ancestrales, espontáneas o de cultivo a lo largo de miles de años, han ocasionado fenómenos de mezcla que han determinado la complejidad de la biología reproductiva de los cítricos cultivados hoy en día. El objetivo global de esta tesis doctoral es aumentar el conocimiento sobre diferentes aspectos reproductivos que son cruciales para la mejora y propagación de los cítricos y su interacción con las condiciones ambientales. El primer objetivo de esta tesis doctoral ha sido el análisis del efecto de la temperatura en la fase progámica de los cítricos. Hemos comprobado que las temperaturas altas redujeron el tiempo que necesitaban los tubos polínicos para llegar a los óvulos y también aceleraron la degeneración del pistilo, mientras que las bajas temperaturas produjeron los efectos opuestos. Sin embargo, los tubos polínicos pudieron llegar a los óvulos en todos los cruces estudiados. Es interesante señalar que observamos por primera vez en cítricos tanto la germinación del polen como el crecimiento del tubo polínico a 10ºC. Las mandarinas representan el 24% de la producción total de cítricos y la ausencia de semillas se valora como criterio de calidad en el mercado de la fruta fresca. La aptitud partenocárpica (AP) es la clave para la producción de frutos sin semillas cuando se combina con la autoincompatibilidad (AI) o la esterilidad. El segundo objetivo de esta tesis doctoral fue evaluar AP y AI para variedades de mandarina con características relevantes como progenitores para la mejora genética de mandarinas sin semillas. Para ello, desarrollamos un protocolo eficiente basado en la emasculación, la autopolinización manual y la polinización cruzada manual. Se observaron seis comportamientos diferentes entre las nueve variedades analizadas. Subrayar que más allá de la importancia crítica de la AI para la producción de frutos sin semillas, la AI es un obstáculo para los programas de mejora genética basados en hibridación, ya que reduce las posibilidades de cruzamiento. Esta circunstancia motivó el planteamiento del tercer objetivo de esta tesis doctoral. Dicho objetivo fue comparar la eficiencia de la ruptura de la reacción de AI por tres factores previamente identificados en otras especies: estrés por temperatura, polinización de yemas florales y poliploidización. Los tres métodos tuvieron éxito en la obtención de plantas autofecundadas, y la polinización de yemas florales resultó ser el método más eficiente. La selección asistida por marcadores (MAS) permite la selección de genotipos en la etapa de plántula, lo que la convierte en una herramienta poderosa, especialmente en especies de árboles con un largo período juvenil, como es el caso de los cítricos. El cuarto objetivo de esta tesis doctoral se centró por tanto en el desarrollo de marcadores SNPs asociados a poliembrionía y esterilidad masculina. Mediante estudios exhaustivos de asociación genética, identificamos una región genómica en el grupo de ligamiento 1 asociado con poliembrionía y una región genómica en el grupo de ligamiento 8 asociado con NPG. Se desarrollaron marcadores SNP cercanos a genes que tienen funciones relacionadas con la esterilidad masculina y con la poliembrionia para las dos regiones genómicas. Esta tesis doctoral proporciona nuevos conocimientos sobre la biología reproductiva de los cítricos y la influencia de la temperatura en ella. El conocimiento generado permitirá implementar de manera más eficiente los programas de mejora genética, particularmente aquellos destinados a la obtención de variedades sin semillas. Además, este conocimiento podrá utilizarse para ayudar en la respuesta a algunos de los desafíos más apremiantes que plantea el escenario actual de calentamiento global. / [CA] Des de que es van originar els cítrics, hibridacions naturals ancestrals, espontànies o de conreu, al llarg de milers d'anys, han ocasionat fenòmens de mescla que han determinat la complexitat de la biologia reproductiva dels cítrics actualment conreats. L'objectiu global d'aquesta tesi va ser augmentar el coneixement de diferents aspectes reproductius dels cítrics ¿especialment en la seua interacció amb les condicions ambientals¿ crucials per a la seua millora genética i reproducció, El nostre primer objectiu ha estat l'anàlisi de l'efecte de la temperatura en la fase progàmica dels cítrics. Hem comprovat que les temperatures càlides redueixen el temps necessari per arribar als òvuls pels tubs de pol·len i que també acceleren la degeneració del pistil; mentre les temperatures fredes van produeixen els efectes contraris. No obstant això, els tubs pol·línics van poder arribar als òvuls en totes les creus estudiades. Curiosament, vam observar per primera vegada als cítrics tant la germinació del pol·len com el creixement del tub pol·línic a 10ºC. Les mandarins representen el 24% de la producció total de cítrics i l'absència de llavors és un criteri de qualitat crucial en el mercat de la fruita fresca. La capacitat partenocàrpica (PA) és la clau per a la producció de fruites sense llavors quan s'acobla a l'autoincompatibilitat (SI) o l'esterilitat. El segon objectiu d'aquesta tesi doctoral va ser avaluar PA i SI per a varietats de mandarines amb característiques rellevants com a progenitors per a la millora genètica de mandarines sense llavors. Amb aquesta finalitat, hem desenvolupat un protocol eficient basat en l'emasculació, l'autopol·linització manual i la pol·linització creuada manual. Es van observar sis comportaments diferents entre les nou varietats analitzades. Subratllar que, enllà de la importància crítica del SI en la producció de fruites sense llavors, el SI és alhora un obstacle per als programes de millora basats en la hibridació, ja que redueix les possibilitats d'encreuament. Aquesta circumstància va motivar el plantejament del tercer objectiu d'aquesta tesi doctoral. Aquest objectiu va ser comparar l'eficiència de la ruptura de la reacció SI per tres factors identificats prèviament en altres espècies: estrès per temperatura, pol·linització de rovells florals i poliploidització. Els tres mètodes van tenir èxit en l'obtenció de plantes autofecundades, i la pol·linització de rovells florals va resultar ser el mètode més eficient. La selecció assistida per marcadors (MAS) en l'etapa de plàntules permet la selecció de genotips diana, la qual cosa la converteix en una eina potent, especialment en espècies d'arbres amb període juvenil llarg, com els cítrics. El quart objectiu d'aquesta tesi doctoral se centra en el desenvolupament de marcadors SNPs associats a la poliembriona i l'esterilitat masculina. Estudis exhaustius d'associació genètica, vam identificar una regió genòmica al grup de lligament 1 associada a la poliembrionia i una regió genòmica al grup de lligament 8 associada amb NPG. Es van desenvolupar marcadors SNP estretament relacionats amb gens implicats per a les dues regions genòmiques. Aquesta tesi doctoral proporciona nous coneixements sobre la biologia reproductiva dels cítrics i la influència de la temperatura sobre ella. El coneixement generat permetrà implementar de manera més eficient els programes de millora genètica, especialment aquells destinats a l'obtenció de varietats sense llavors. A més, aquest coneixement es podrà utilitzar en la complexa resposta a alguns dels reptes més urgents que planteja l'escenari actual d'escalfament global. / [EN] Since the origin of citrus, ancestral natural hybridizations, thousands of years of cultivation and admixture phenomenon have resulted in the complexity of the reproductive biology of today's cultivated citrus. The global objective of this PhD was to increase our knowledge on different reproductive aspects that are crucial for citrus breeding and propagation and their interaction with environmental conditions. Temperature in the progamic phase is critical for the success of plant sexual reproduction. Pollen grain germination, stigmatic receptivity and pollen tube growth are the main components of this phase. The analysis of temperature effect on the progamic phase of citrus was our first objective. Using three compatible crosses within the Citrus genus, we evaluated, with this method, the effect of four temperature regimes in every process during the progamic phase. Warm temperatures reduced the time needed by pollen tubes to reach the ovules and also accelerate pistil degeneration while cold temperatures produced the opposite effects. However, pollen tubes were able to reach the ovules in all crosses studied. Interestingly, we observed for the first time in citrus both pollen germination and pollen tube growth at 10ºC. Mandarins account for 24% of total citrus production and seedlessness is a crucial quality criterion for the mandarin fresh fruit market. Parthenocarpic ability (PA) is the key for seedless fruit production when it is coupled with self-incompatibility (SI) or sterility. The second objective of this PhD dissertation was to evaluate PA and SI for mandarin varieties with relevant characteristics as parents for seedless mandarin breeding. For this purpose, we developed an efficient protocol based on emasculation, hand self-pollination and hand cross-pollination. Pollen performance was analysed by histological observations, together with fruit set and seed production. Six different behaviors were observed among the nine varieties analysed. Beyond the critical importance of SI for seedless fruit production, SI is an obstacle for breeding programs based on hybridization as it reduces crossing possibilities. The third objective of this PhD dissertation was to compare the efficiency of the SI reaction breakdown by three factors previously identified in other plant species: temperature stress, bud pollination and polyploidization. The three methods were successful in recovering selfed plants, and bud pollination was the most efficient approach. Chromosome doubling was also efficient, but involved developing tetraploid plants. Cold temperature stress allowed us to obtain a few diploid selfed plants. However, this method proved much more complex to apply than bud pollination in specific breeding programs. Marker-assisted selection (MAS) allows the selection of target genotypes at the seedling stage, making it a powerful tool, especially in tree species with long juvenile period, such as citrus. The 4th objective of this PhD dissertation focuses on the development of SNPs markers associated with polyembryony and male sterility. Through genetic association studies, we identified a genomic region in linkage group 1 associated with polyembryony and a genomic region on linkage group 8 associated with NPGA. SNP markers closely linked with implied genes were developed for the two genomic regions. Overall, this PhD dissertation provides new insights into citrus reproductive biology and the influence of temperature on it. The knowledge generated will allow to implement breeding programs more efficiently, particularly those aimed at obtaining seedless varieties. This knowledge can eventually be used to respond to some of the most pressing challenges posed by the current global warming scenario. / Montalt Resurrección, R. (2023). Assessment of Citrus Reproductive Biology for Seedless Mandarin Production and its Interaction with Temperature [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/193856 / Compendio

In this dissertation we explore the concept that the works of God continue in a creative and coherent continuum from the portrayal in Genesis 1 to the description of their consummation in Revelation through the dynamic broad movement of the Kingdom of God that moved through Israel and the other nations of the world and through the churches and other institutions as well as in the Bible, in the experiences of humanity and in the vast non-human creation. Therefore creation, redemption, renewal and consummation are seen to be linked in a process which is not confined to the usual theological pointers of the Church and the Bible. Churches and the Bible are signs and instruments of the Kingdom of God amongst and in the many universes and not the only signs and instruments of the Kingdom of God in and amongst the many universes. The importance of the coherence between each work of God, the otherness of each work and the ongoing character of God's works demonstrates itself in the impact of faith on human experience and on the very existence of human beings. The concept of developing a wholesome person with a fivesome awareness is developed, this being an awareness of a human beings creatureliness, an awareness of their sinful tendencies of doing damage to God, the self as being created by God and all other creatures of God, an awareness of the salvific and reconciliatory power of the cross and resurrection of Jesus Christ and an awareness of the renovating and renewal power of the Spirit of Pentecost which is carrying and guiding the process of the fivesome awareness in our minds, our experiences and our living into afterlife towards the consummation of all things in the new heaven and the new earth. The ongoing experience of anticipatory fragments and moments of meaning and significance breaking in from the future into our current life, bridges God's Kingdom works in the future through reflective understanding of faith experience with our everyday experience as human beings. The basic point and the thrust of this dissertation is that of a `both...and' and not an `either...or' relationship between human beings and their experience of God as Creator, their own falling into sin, God as Reconciliator in Christ, God the Holy Spirit as the Renewer and Renovator of all things unto afterlife and the creation of the new heaven and the new earth - the whole ongoing process of God's works is attested in the Biblical texts and attested in the foursome and inclusive experiential awareness of God's works in our daily experience by the Holy Spirit. / Systematic Theology and Theological Ethics / M.Th. (Systematic Theology)

The aim of this thesis is to compare the concept of sin in the theologies of Ellen G. White and Leonardo Boff. Chapter 1 examines Ellen G. White's concept of sin. White's historical and theological backgrounds coupled with her use of the "great controversy" motif provide a better grasp of her understanding of sin. White defines sin as the transgression of God's Law. She views sin to comprise at least two dimensions, namely, the individual and the social. White regards these two aspects of sin to have equal significance. White's detailed treatment of the sanctuary teaching also highlights the two dimensions of sin. In Chapter 2 Boff' s idea of sin is investigated. Boff' s historical background, which exposed him to the poor, influenced his perception of sin. Boff's theological background together with his familiarity with Karl Marx's social analysis prompted Boff to define sin as the negation of God's love in a human history bedevilled by class conflict. Boff views sin to have the individual and social dimensions. Yet, in terms of importance, Boff believes that the social dimension of sin is more consequential than the individual one. In Chapter 3 White's and Boff s views on sin are compared. From this comparison it is evident that both White and Boff recognize the bipolarity of sin. Both seem to agree that christians should take an active role in correcting social evils because love for God is manifested by how we relate to our neighbor. Boff devotes less space to the individual aspect of sin than White. Chapter 4 shows that White's theological tradition has a lot to learn from Boff and his tradition and also vice versa. An awareness of the current priestly ministry of Christ evident in White's theology could help Boff to bring some balance to his stance on the social and the individual dimension of sin. Boff' s use of Marx's social analysis should also help Seventh-day Adventists, the inheritors of White's theology, not to interpret White's theology of sin only along individualistic lines while overlooking its social dimension. / Philosophy, Practical & Systematic Theology / D.Th. (Systematic Theology)

German text / Die vorliegende Arbeit schlägt eine Brücke über den Graben zwischen dem normativ-ethischen Anspruch an die Lebenslänglichkeit einer Ehe und Gottes gnädigem Handeln gegenüber dem scheiternden Menschen. Anhand des Umgangs der Kirchen und Glaubensgemeinschaften mit Ehescheidung und Wiederheirat wird dieser Graben aufgezeigt. Es ergibt sich, dass die bestehende Spannung jeweils durch die einseitige Betonung von Norm oder Gnade aufgelöst wird. Die Grundlage für die Diskussion bilden das massgebende Eheverständnis sowie die Bestimmung des Verhältnisses zwischen theologischer Ethik und dem Konzept der Gnade als Handlungsprinzip Gottes. Darum werden in dieser Forschungsarbeit zuerst die theologiegeschichtlich gewachsenen Eheverständnisse und die unterschiedlichen Normen- und Gnadenverständnisse der römisch katholischen Kirche, der evangelischen Kirchen und der evangelikalen Gemeinschaften in Bezug auf den Umgang mit Scheidung und Wiederheirat untersucht. Aufgrund der herausgearbeiteten Übereinstimmungen und Unterschiede wird anschliessend anhand eines von der Vergebung ausgehenden Denkansatzes der Graben zwischen normativer Ethik und dem Konzept der Gnade überbrückt, die Spannung aber nicht aufgehoben. Daraus werden mögliche Auswirkungen im Umgang mit Scheidung und Wiederheirat für die Kirchen und Gemeinschaften abgeleitet. / The present thesis bridges the gap between the normative-ethical standard of lifelong marriage on the one hand and God’s grace for men‘s failures on the other hand. Considering the practices of churches and communities with regard to divorce and remarriage, this gap is demonstrated. It is found that the associated tension is eliminated by an unilateral choice of either the normative aspect or the principle of grace. The basis for the discussion is provided by the relevant understanding of marriage and the determination of the relationship between theological ethics and grace as the principle of God's action towards mankind. Therefore, this thesis investigates the evolution of the theological understanding of marriage during history and the different understandings of norms and grace in the Catholic Church, the Protestant Church and the Evangelical Free Communities with regard to their handling of divorce and remarriage. Considering the resulting similarities and differences, I propose – building on the fact of God’s forgiveness – how to bridge the gap between normative ethics and the concept of grace without eliminating the obvious tension. Finally, possible consequences for how to handle divorce and remarriage by churches and communities are drawn. / Philosophy & Systematic Theology / M. Th. (Theological Ethics)

Vetores lentivirais são ferramentas fundamentais para modificação celular. Sua utilização ganhou destaque devido à capacidade desses em integrar ao genoma de células que estão ou não em divisão. Grande parte dos vetores desenvolvidos são derivados do genoma do Vírus da Imunodeficiência Humana (HIV-1), portanto, modificações foram necessárias a fim de evitar a formação de Partículas Competentes em Replicação (RCLs) e garantir uma utilização segura. Com as modificações, foram produzidos os vetores lentivirais de terceira geração utilizados atualmente. Esses vetores podem ser usados para expressão constitutiva de genes, produção de proteínas recombinantes, produção de animais transgênicos e terapia gênica. Com isso, torna-se necessário o desenvolvimento de vetores lentivirais para aplicação em pesquisa básica e ensaios clínicos. Dessa forma, o presente estudo teve por objetivo a construção de vetores de expressão lentivirais aplicáveis à: 1- expressão constitutiva de genes de interesse e 2-vetores com promotores específicos para expressão de proteínas em megacariócitos. Esse trabalho descreve a construção desses vetores, sua importância e discute suas possíveis aplicações. As sequências selecionadas para produção dos vetores foram: os genes Runx1C e VkorC1 e os promotores proPF4 e proITGA2b. Todas as sequências encontram-se clonadas em vetor de clonagem e estoques de bactérias com esses vetores congeladas em glicerol foram confeccionados. Para a confecção dos vetores lentivirais, o gene Runx1C foi subclonado no vetor lentiviral base p1054-CIGWS sob controle do promotor forte CMV, enquanto o promotor proITGA2b foi subclonado no vetor base p1054-FVIII, em substituição ao promotor CMV, de forma a controlar a expressão de FVIII. Os dois vetores produzidos apresentam ainda o gene para proteína verde GFP precedida do sítio de ligação do ribossomo IRES, com expressão controlada pelo mesmo promotor interno do vetor. O trabalho possibilitou, portanto, a produção de dois vetores lentivirais bi-cistrônicos: p1054-Runx1C e pL-proITGA2b-FVIII. A construção p1054-Runx1C ainda não foi sequenciada, mas foi confirmada por restrição enzimática e apresenta potencial para aplicação em estudos de diferenciação hematopoética. Já a construção pL-proITGA2b-FVIII foi sequenciada, porém sem confirmação da região de ligação do proITGA2b ao vetor. Reações de PCR e de restrição enzimática confirmaram a ligação e sequenciamento mostrou 67% de similaridade entre a região sequenciada e o promotor ITGA2b depositado no banco de dados. Análise funcional foi realizada através da transfecção desse vetor em células HEK-293T. As células transfectadas apresentaram expressão positiva para GFP e secreção de FVIII no sobrenadante celular, evidenciando que o promotor proITGA2b clonado no vetor encontra-se ativo. Esse vetor apresenta potencial para aplicação em terapia gênica para hemofilias, pois apresenta expressão do fator de coagulação direcionado a megacariócitos e plaquetas, células que estão diretamente relacionadas ao processo de coagulação, representando grandes veículos para secreção desses fatores. Ainda, os dois vetores lentivirais gerados apresentam segurança e eficiência elevadas, pois são vetores de terceira geração auto-inativantes (SIN) e apresentam elementos regulatórios que melhoram o transporte e integração do DNA ao genoma hospedeiro. / Lentiviral vectors are fundamental tools for cell modification that gained prominence due to their ability to integrate the genome of non-dividing cells. Most of developed lentiviral vectors are derived from the genome of Human Immunodeficiency Virus (HIV-1), so modifications were necessary in order to avoid the formation of Competent Replication Particles (RCLs) and ensure safer operations. The modifications led to development of third generation lentiviral vectors currently used. These vectors can be used for constitutive gene expression, production of recombinant protein, production of transgenic animals and gene therapy. It\'s evident the need to develop lentiviral vectors for application in basic research and clinical trials. Thus this study aimed to construct lentiviral expression vectors applicable to: 1- constitutive expression of genes of interest and 2-vectors with specific promoters for expression of proteins in megakaryocytes and platelets. This paper describes the construction of these vectors, their importance and discuss their possible applications. Sequences were selected for production of the vectors: genes Runx1C and VkorC1 and proPF4 and proITGA2b promoters. All four sequences are cloned into cloning vectors and stocks of bacteria with these vectors frozen in glycerol were prepared. Lentiviral vectors were engineered from subcloning the sequence Runx1C into the basic lentiviral vector p1054- CIGWS under control of the strong CMV promoter, and from subcloning proITGA2b promoter into p1054-FVIII basic vector, replacing the CMV promoter in order to control the expression of FVIII. Both vectors exhibit the green fluorescence protein GFP gene preceded by a ribosome binding site IRES under control of vector\'s internal promoter. Therefore, this work resulted in the production of two bi-cistronic lentiviral vectors: p1054-Runx1C and pLproITGA2b-FVIII. The p1054-Runx1C construction has not yet been sequenced, but it was confirmed by digestion and has potential for use in hematopoietic differentiation studies. Though, pL-proITGA2b-FVIII construct was sequenced, but the technique didn\'t allow to confirm the binding region between proITGA2b and the vector. Although PCR reaction and digestion confirmed the construction. Sequence analysis showed 67% similarity between the sequenced region and ITGA2b promoter deposited in the database. Functional analysis was performed by transfection of this vector in HEK-293T cells. The transfected cells showed positive expression of GFP and FVIII secretion in cell supernatant, indicating that the proITGA2b promoter cloned into the vector is active. This vector has potential usage in gene therapy for hemophilia, since it can be used to express coagulation factors in megakaryocytes and platelets and these cells are directly related to the clotting process, representing great vehicles for secretion of these factors. Even more, the two lentiviral vectors generated have higher safety and efficiency, as they are self-inactivating (SIN) third-generation vectors and have regulatory elements that enhance transport and integration of DNA into the host genome.

Este trabalho se debruça sobre a poesia do exílio republicano espanhol de 1939. A pesquisa procura demonstrar que o tempo de duração do exílio é fator preponderante para a leitura de uma produção poética ampla e dispersa. Desse modo, seu enfoque central trata de investigar como essa poética exilada assimilou e ressignificou as três décadas de exílio, reconstruindo a identidade de seus autores. O corpus é composto por livros de inflexão da obra de três autores: Luis Cernuda (Vivir sin estar viviendo, 1944-1949), Emilio Prados (La piedra escrita, 1959-1961) e Max Aub (Antología traducida, 1963-1971). / The present study examines the 1939 Spanish Republican exile poetry. The investigation aims to demonstrate that the time the exile lasted is a fundamental aspect when reading such a widespread poetic production. Thus, the focus is on analyzing in which way this exiled poetry incorporated and resignified the three decades of exile, reconstructing the authors identity. Critical books in three authors works compose the corpus: Luis Cernuda (Vivir sin estar viviendo, 1944-1949), Emilio Prados (La piedra escrita, 1959- 1961) and Max Aub (Antología traducida, 1963-1971).

In dieser Arbeit untersuchen wir die grundlegenden Wachstums- und Relaxationsprozesse, die es erlauben den Verzerrungszustand von GaN (0001) beim Wachstum auf Silizium (111) Substraten einzustellen und die resultierende Dichte an Durchstoßversetzungen zu reduzieren. Zu deren Analyse werden GaN (0001) Schichten, die mittels metallorganischer Gasphasenepitaxy abgeschieden worden sind, hauptsächlich mit transmissionselekronenmikroskopischen Methoden untersucht. Die wesentlichen Erkenntnisse der Arbeit sind: (i) Der Aufbau einer kompressiven Verzerrung von GaN (0001) Filmen mittels AlGaN Zwischenschichten beruht auf einer Asymmetrie der plastischen Relaxation an den beiden Grenzflächen der AlGaN Zwischenschicht. Fehlpassungsversetzungen bilden sich zwar an beiden Grenzflächen aus, jedoch ist der mittlere Abstand zwischen Versetzungslinien an der unteren Grenzfläche kleiner, als an der oberen. (ii) Plastische Relaxation von verzerrten (0001) Wurtzit Schichten erfolgt im Wesentlichen durch Bildung von a-Typ Fehlpassungsversetzungen im 1/3 |{0001} Gleitsystem. Diese bilden sich aber nur dann, wenn die verzerrten Schichten eine 3-D Morphologie aufweisen. Eine quantitative Modellierung dieses Prozesses zeigt, dass die kritische Schichtdicke für das Einsetzen der plastischen Relaxation wesentlich vom Wachstumsmodus bestimmt wird. (iii) Eine Silizium Delta-Dotierung der GaN (0001) Oberfläche führt zum Wachstum einer kohärenten Sub-Monolage SiGaN3, die eine periodisch Anordnung von Silizium- und Galliumatomen, sowie Galliumvakanzen aufweist. Da das Wachstum von GaN direkt auf der SiGaN3-Monolage unterdrückt ist, tritt ein Übergang zu 3-D Inselwachstum auf, das zunächst ausschließlich in Löchern der SiGaN3-Monolage anfängt. Eine hohe Konzentration von Silizium auf der GaN (0001) Oberfläche wirkt also als Anti-Surfactant beim epitaktischen Wachstum von GaN. Rechnungen mittels der Dichtefunktionaltheorie liefern Erklärungen für das beobachtete Wachstumsverhalten. / In this work we study the basic growth and relaxation processes that are used for strain and dislocation engineering in the growth of GaN (0001) films on silicon (111) substrates. To analyse these processes, samples, grown by metalorganic vapour phase epitaxy were investigate by means of transmission electron microscopy. Our investigations have revealed the following main results: (i) Strain engineering and build-up of compressive strain in GaN (0001) films by means of AlGaN interlayer is based on an asymmetry in plastic relaxation between the two interfaces of the AlGaN interlayer. Although misfit dislocation networks form at both interfaces of the interlayer, the average spacing of dislocation lines at the lower interface is smaller than that at the upper one. (ii) Plastic relaxation of strained (0001) wurtzite films is caused mainly by formation of a-type misfit dislocations in the 1/3 |{0001} slip-system. These a-type misfit dislocations form once the strained films undergo a transition to a 3-D surface morphology, e.g. by island growth or cracking. Quantitative modelling of this process reveals that the critical thickness for nucleation of a-type misfit dislocations depends next to the lattice mismatch mainly on the growth mode of the film. (iii) Silicon delta-doping of the GaN (0001) surface leads to the growth of a coherent sub-monolayer of SiGaN3 that shows a periodic arrangement of silicon and gallium atoms and gallium vacancies. Since growth of thick GaN layers directly on top of the SiGaN3-monolayer is inhibited a transition towards 3-D island growth occurs, whereby GaN islands exclusively nucleate at openings in the SiGaN3-monolayer. A high concentration of silicon on the GaN (0001) surface thus acts as an anti-surfactant in the epitaxial growth of GaN. Our density functional theory calculations provide an explanation for both the self-limited growth of the SiGaN3-monolayer, as well as for the blocking of GaN growth on top of the SiGaN3-monolayer.

Vetores lentivirais são ferramentas fundamentais para modificação celular. Sua utilização ganhou destaque devido à capacidade desses em integrar ao genoma de células que estão ou não em divisão. Grande parte dos vetores desenvolvidos são derivados do genoma do Vírus da Imunodeficiência Humana (HIV-1), portanto, modificações foram necessárias a fim de evitar a formação de Partículas Competentes em Replicação (RCLs) e garantir uma utilização segura. Com as modificações, foram produzidos os vetores lentivirais de terceira geração utilizados atualmente. Esses vetores podem ser usados para expressão constitutiva de genes, produção de proteínas recombinantes, produção de animais transgênicos e terapia gênica. Com isso, torna-se necessário o desenvolvimento de vetores lentivirais para aplicação em pesquisa básica e ensaios clínicos. Dessa forma, o presente estudo teve por objetivo a construção de vetores de expressão lentivirais aplicáveis à: 1- expressão constitutiva de genes de interesse e 2-vetores com promotores específicos para expressão de proteínas em megacariócitos. Esse trabalho descreve a construção desses vetores, sua importância e discute suas possíveis aplicações. As sequências selecionadas para produção dos vetores foram: os genes Runx1C e VkorC1 e os promotores proPF4 e proITGA2b. Todas as sequências encontram-se clonadas em vetor de clonagem e estoques de bactérias com esses vetores congeladas em glicerol foram confeccionados. Para a confecção dos vetores lentivirais, o gene Runx1C foi subclonado no vetor lentiviral base p1054-CIGWS sob controle do promotor forte CMV, enquanto o promotor proITGA2b foi subclonado no vetor base p1054-FVIII, em substituição ao promotor CMV, de forma a controlar a expressão de FVIII. Os dois vetores produzidos apresentam ainda o gene para proteína verde GFP precedida do sítio de ligação do ribossomo IRES, com expressão controlada pelo mesmo promotor interno do vetor. O trabalho possibilitou, portanto, a produção de dois vetores lentivirais bi-cistrônicos: p1054-Runx1C e pL-proITGA2b-FVIII. A construção p1054-Runx1C ainda não foi sequenciada, mas foi confirmada por restrição enzimática e apresenta potencial para aplicação em estudos de diferenciação hematopoética. Já a construção pL-proITGA2b-FVIII foi sequenciada, porém sem confirmação da região de ligação do proITGA2b ao vetor. Reações de PCR e de restrição enzimática confirmaram a ligação e sequenciamento mostrou 67% de similaridade entre a região sequenciada e o promotor ITGA2b depositado no banco de dados. Análise funcional foi realizada através da transfecção desse vetor em células HEK-293T. As células transfectadas apresentaram expressão positiva para GFP e secreção de FVIII no sobrenadante celular, evidenciando que o promotor proITGA2b clonado no vetor encontra-se ativo. Esse vetor apresenta potencial para aplicação em terapia gênica para hemofilias, pois apresenta expressão do fator de coagulação direcionado a megacariócitos e plaquetas, células que estão diretamente relacionadas ao processo de coagulação, representando grandes veículos para secreção desses fatores. Ainda, os dois vetores lentivirais gerados apresentam segurança e eficiência elevadas, pois são vetores de terceira geração auto-inativantes (SIN) e apresentam elementos regulatórios que melhoram o transporte e integração do DNA ao genoma hospedeiro. / Lentiviral vectors are fundamental tools for cell modification that gained prominence due to their ability to integrate the genome of non-dividing cells. Most of developed lentiviral vectors are derived from the genome of Human Immunodeficiency Virus (HIV-1), so modifications were necessary in order to avoid the formation of Competent Replication Particles (RCLs) and ensure safer operations. The modifications led to development of third generation lentiviral vectors currently used. These vectors can be used for constitutive gene expression, production of recombinant protein, production of transgenic animals and gene therapy. It\'s evident the need to develop lentiviral vectors for application in basic research and clinical trials. Thus this study aimed to construct lentiviral expression vectors applicable to: 1- constitutive expression of genes of interest and 2-vectors with specific promoters for expression of proteins in megakaryocytes and platelets. This paper describes the construction of these vectors, their importance and discuss their possible applications. Sequences were selected for production of the vectors: genes Runx1C and VkorC1 and proPF4 and proITGA2b promoters. All four sequences are cloned into cloning vectors and stocks of bacteria with these vectors frozen in glycerol were prepared. Lentiviral vectors were engineered from subcloning the sequence Runx1C into the basic lentiviral vector p1054- CIGWS under control of the strong CMV promoter, and from subcloning proITGA2b promoter into p1054-FVIII basic vector, replacing the CMV promoter in order to control the expression of FVIII. Both vectors exhibit the green fluorescence protein GFP gene preceded by a ribosome binding site IRES under control of vector\'s internal promoter. Therefore, this work resulted in the production of two bi-cistronic lentiviral vectors: p1054-Runx1C and pLproITGA2b-FVIII. The p1054-Runx1C construction has not yet been sequenced, but it was confirmed by digestion and has potential for use in hematopoietic differentiation studies. Though, pL-proITGA2b-FVIII construct was sequenced, but the technique didn\'t allow to confirm the binding region between proITGA2b and the vector. Although PCR reaction and digestion confirmed the construction. Sequence analysis showed 67% similarity between the sequenced region and ITGA2b promoter deposited in the database. Functional analysis was performed by transfection of this vector in HEK-293T cells. The transfected cells showed positive expression of GFP and FVIII secretion in cell supernatant, indicating that the proITGA2b promoter cloned into the vector is active. This vector has potential usage in gene therapy for hemophilia, since it can be used to express coagulation factors in megakaryocytes and platelets and these cells are directly related to the clotting process, representing great vehicles for secretion of these factors. Even more, the two lentiviral vectors generated have higher safety and efficiency, as they are self-inactivating (SIN) third-generation vectors and have regulatory elements that enhance transport and integration of DNA into the host genome.

Descripción y análisis del fenómeno del homeschooling en España desde la doble perspectiva sincrónica (circunscrita al año 2009) y diacrónica (desde sus comienzos hasta nuestros días). Esta descripción y análisis se estructura en cuatro grandes apartados o capítulos. En el primero de ellos, se fijan los objetivos del trabajo; se establece la metodología; se delimitan los conceptos más frecuentemente utilizados en el estudio; se define el fenómeno; se informa de las diferentes denominaciones con las que se conoce esta práctica; y, finalmente se ofrece un panorama de la situación en la que se encuentra el homeschooling en el mundo (países en los que está oficialmente reconocido). En el segundo capítulo, se realiza una descripción histórica de los principales acontecimientos del homeschooling español, desde sus comienzos hasta 2012. En el tercer capítulo se ofrecen los resultados obtenidos en las dos encuestas realizadas sobre el homeschooling en España. En el cuarto y último capítulo, se aborda el homeschooling español desde una perspectiva cualitativa.

In this dissertation we explore the concept that the works of God continue in a creative and coherent continuum from the portrayal in Genesis 1 to the description of their consummation in Revelation through the dynamic broad movement of the Kingdom of God that moved through Israel and the other nations of the world and through the churches and other institutions as well as in the Bible, in the experiences of humanity and in the vast non-human creation. Therefore creation, redemption, renewal and consummation are seen to be linked in a process which is not confined to the usual theological pointers of the Church and the Bible. Churches and the Bible are signs and instruments of the Kingdom of God amongst and in the many universes and not the only signs and instruments of the Kingdom of God in and amongst the many universes. The importance of the coherence between each work of God, the otherness of each work and the ongoing character of God's works demonstrates itself in the impact of faith on human experience and on the very existence of human beings. The concept of developing a wholesome person with a fivesome awareness is developed, this being an awareness of a human beings creatureliness, an awareness of their sinful tendencies of doing damage to God, the self as being created by God and all other creatures of God, an awareness of the salvific and reconciliatory power of the cross and resurrection of Jesus Christ and an awareness of the renovating and renewal power of the Spirit of Pentecost which is carrying and guiding the process of the fivesome awareness in our minds, our experiences and our living into afterlife towards the consummation of all things in the new heaven and the new earth. The ongoing experience of anticipatory fragments and moments of meaning and significance breaking in from the future into our current life, bridges God's Kingdom works in the future through reflective understanding of faith experience with our everyday experience as human beings. The basic point and the thrust of this dissertation is that of a `both...and' and not an `either...or' relationship between human beings and their experience of God as Creator, their own falling into sin, God as Reconciliator in Christ, God the Holy Spirit as the Renewer and Renovator of all things unto afterlife and the creation of the new heaven and the new earth - the whole ongoing process of God's works is attested in the Biblical texts and attested in the foursome and inclusive experiential awareness of God's works in our daily experience by the Holy Spirit. / Systematic Theology and Theological Ethics / M.Th. (Systematic Theology)