A variação do número de cópias (CNV) de genes e cromossomos é uma característica comum do genoma plástico de Leishmania major, que pode estar associada à resistência do parasita à quimioterapia das leishmanioses. Em outros eucariotos, alterações na replicação do DNA ou na resposta a danos no DNA (DDR) pode levar à CNV. Nestes organismos, o complexo de checkpoint 9-1-1 (RAD9, RAD1 e HUS1) é essencial para a detecção e a sinalização do estresse de replicação e para o recrutamento de uma apropriada DDR. Já demonstramos que L. major expressa um homólogo 9-1-1 funcional. Aqui, avaliamos a deficiência de subunidades de 9-1-1 na variação do número de cópias em células selecionadas em metotrexato (MTX), um inibidor da enzima diidrofolato redutase timidilato sintetase (DHFR-TS). A seleção em MTX facilita o isolamento de células que carregam amplificações contendo o locus da DHFR-TS. Assim, selecionamos células deficientes de HUS1 ou RAD9 para resistência ao MTX sem e com exposição previa a hidroxiureia (HU), uma droga que causa estresse de replicação por inibição da ribonucleotídeo redutase, e avaliamos o efeito da deficiência destas proteínas na CNV e no tipo de amplificação gerada. Avaliamos também o efeito da deficiência destas proteínas no processo de síntese do DNA medido pela incorporação de IdU e observamos que a deficiência destas proteínas levou a um incremento na síntese do DNA na ausência de estresse de replicação e a perfis opostos de síntese do DNA após a remoção do estresse replicativo. Análises da detecção de simples fita do DNA (ssDNA) e da histona H2A fosforilada (?H2A) como indicadores do processo de estresse de replicação e dano no DNA também foram conduzidas. Em conjunto, nossos resultados indicam que (i) os níveis alterados das proteínas HUS1 e RAD9 afetam o padrão da CNV após a seleção no MTX, assim como a natureza da amplificação; (ii) HUS1 e RAD9 parecem possuir mecanismos distintos para mediar a CNV; (iii) a função destas proteínas na CNV deve envolver o processo de replicação e (iv) HUS1 e RAD9 são requeridas para a manutenção da estabilidade genômica em Leishmania. Estes resultados contribuem para uma melhor compreensão não só da evolução da via de sinalização mediada pelo complexo de checkpoint 9-1-1 nos eucariotos, mas também da bases moleculares da plasticidade genômica e do fenômeno de amplificação gênica em Leishmania. / The copy number variation (CNV) of genes and chromosomes is a common feature of the plastic genome of Leishmania major, which is normally associated with resistance of the parasite to the chemotherapy of leishmaniasis. In other eukaryotes, alteration in DNA replication and DNA damage response (DDR) causes CNV. In these organisms, the RAD9-RAD1-HUS1 (9-1-1) checkpoint complex is essential for detection and signaling of replication stress and recruitment of an appropriate DDR. We have already demonstrated that L. major expresses a functional 9-1-1 homolog. Here we evaluated the effect of 9-1-1 subunit deficiency in CNV of cells selected in methotrexate (MTX), an inhibitor of the dihydrofolate reductase thymidylate synthetase (DHFR-TS) enzyme. Selection in MTX facilitates the isolation of cells that carry amplicons containing the DHFR-TS locus. Thus, we selected HUS1 or RAD9 deficient cells for MTX resistance without and prior exposure to hydroxyurea (HU), a drug that causes replication stress due to inhibition of ribonucleotide reductase, and evaluated not only CNV, but also the nature of the amplification generated. We also evaluated the effect of deficiency of these proteins in the DNA synthesis process measured by IdU incorporation and observed that the deficiency of these proteins led to an increase in DNA synthesis in the absence of replication stress, and to opposite profiles of DNA synthesis after removal of replicative stress. Analyzes of single-stranded DNA (ssDNA) and phosphorylated histone H2A (?H2A) as indicators of replication stress and DNA damage were also conducted in both presence and absence of replicative stress. Taken together, our results indicate that (i) altered levels of HUS1 and RAD9 proteins affect the CNV pattern after selection in MTX, as well as the nature of amplification; (ii) HUS1 and RAD9 possibly have different mechanisms to mediate CNV; (iii) the function of these proteins in CNV seems to involve replication process and (iv) HUS1 and RAD9 are required for the maintenance of genomic stability in Leishmania. These findings contribute to a better understanding not only of the evolution of the signaling pathway mediated by 9-1-1 checkpoint complex in eukaryotes, but also of the molecular basis of the genome plasticity and the gene amplification phenomenon in Leishmania.

A estabilidade genômica é condição essencial à sobrevivência e ao funcionamento dos organismos vivos. No entanto, várias situações podem provocar danos no DNA. Por exemplo, cerca de 104 lesões podem ocorrer no material genético de uma célula de mamífero a cada dia. No intuito de preservar a integridade genômica e contornar os efeitos deletérios destas modificações, uma maquinaria constituída de proteínas especializadas em reconhecer e reparar estes danos foi selecionada ao longo do curso evolutivo. Defeitos em proteínas destas maquinarias causam instabilidade genômica e pode resultar em elevada taxa de mutações e quebras do DNA que resultam em eventos de amplificação gênica, como em células cancerosas. De uma maneira aparentemente contrária ao requerimento de estabilidade genômica como condição primordial para a perpetuação da vida, Leishmania apresenta um genoma notavelmente maleável e explora a amplificação gênica como recurso de sobrevivência. Ainda que a plasticidade genômica em Leishmania seja facilmente demonstrada, nós não conhecemos os mecanismos precisos pelos quais este parasita coordena a ação da maquinaria de detecção de danos no DNA e a consumação dos eventos de amplificação gênica. No intuito de contribuir para a compreensão deste processo, nós identificamos proteínas homólogas do complexo 9-1-1 (Rad9-Hus1-Rad1) em Leishmania major. As proteínas LmHus1 e LmRad9 apresentam marcada divergência estrutural em relação aos seus homólogos em outros eucariotos e nenhuma proteína obviamente homóloga a Rad1 foi identificada neste parasita. Análises filogenéticas indicam que LmHus1 e LmRad9 são relacionadas ao complexos heterotriméricos envolvidos na detecção de danos no DNA. Em acordo com isso, nossos experimentos demonstram que alteração nos níveis destas proteínas interfere na capacidade do parasita em lidar com estresse genotóxico. LmHus1 localiza-se no núcleo, é requerida para o crescimento normal deste parasita e a diminuição de sua expressão compromete mecanismos de controle de ciclo celular e manutenção de telômeros. LmRad9 também localiza-se no núcleo e sua superexpressão causa defeito de crescimento e de resposta ao estresse genotóxico em L. major. Nós observamos que LmHus1 e LmRad9 formam um complexo responsivo ao dano no DNA in vivo, uma forte indicação de que o complexo 9-1-1 tenha sido conservado em L. major. As peculiaridades estruturais destas proteínas sugerem que o complexo 9-1-1 de L. major possua uma arquitetura distinta em comparação aos eucariotos superiores. Em adição a isto, outras proteínas, tais como a LmRpa1, também apresentam uma marcante divergência estrutural. Isso sugere que a via de sinalização de danos no DNA envolvendo o complexo 9-1-1 e Rpa1 de L. major possua mecanismos peculiares de ação. Estas observações podem permitir entender como ocorreu o processo evolutivo da sinalização mediada pelo complexo 9-1-1 nos eucariotos, além de ajudar para o entendimento das bases moleculares de como este parasito conduz os eventos de amplificação gênica. / Genome stability is a essential condition for survival and proper functioning of living organisms. However, a broad range of elements may lead to DNA damage. For instance, about 104 DNA lesions may be inflicted upon any given mammalian cell everyday. In order to maintain the genome integrity and circumvent the deleterious effects of these lesions, a molecular machinery composed of proteins specialized in detecting and repairing DNA damage has been selected in evolution. Defects of the proteins that constitute such machineries may result not only in a high mutation rate, but also in breaks in the DNA structure that can mediate gene amplification as observed in cancer cells. In an apparent opposition to such requirement for stability as an essential condition to life, the protozoan Leishmania presents a highly malleable genome and explores genome amplification as a survival and adaptation tool. Despite of the fact that the Leishmania genome plasticity can be easily demonstrated, the precise mechanisms that coordinate the molecular machineries involved in the detection and signaling of DNA damage, and in the regulation of gene amplification is still largely unknown. In order to contribute to a better understanding of these processes, we identified and studied the Leishmania major proteins that are homologues of those proteins that compose the 9-1-1 complex (Rad9-Hus1-Rad1). The proteins LmHus1 and LmRad9 present a high structural divergence when compared to its homologues from other eukaryotes and no obvious homologue of Rad1 was identified in the parasite genome. Phylogeny analysis indicated that LmHus1 and LmRad9 are closely related to heterotrimeric complexes involved in the detection of DNA damage. In accordance to that, our experiments demonstrated that altered levels of these proteins interfere with the parasite ability to deal with genotoxic stress. Moreover, LmHus1 was localized to the parasite nucleus and is a required protein for normal parasite proliferation. Besides, we showed that decreased levels of LmHus1 compromise cell cycle regulation and the maintenance of telomeres. LmRad9 was also shown to be localized to the cell nucleus and its overexpression led to growth defects and affected the L. major response to genotoxic stress. We also observed that LmHus1 and LmRad9 interact with each other to for a protein complex that is responsive to DNA damage in vivo, which strongly suggested that the 9-1-1 complex was conserved in L. major. The structural peculiarities of these proteins indicate that the possible L. major 9-1-1 complex has a different architecture when compared to the complex found in higher eukaryotes. In addition to that, other proteins, such as LmRpa1, also present a marked structural divergence. Altogether, these findings suggest that the DNA damage signaling pathway involving the 9-1-1 complex and LmRpa1 in L. major, may present a peculiar mode of action. These observations may contribute to a better understanding not only of the evolution of the signaling pathway mediated by the 9-1-1 complex in eukaryotes, but also of the molecular basis of the genome plasticity and the gene amplification phenomenon.

Este trabalho tem como objetivo apresentar um estudo detalhado dos fundamentos da Geometria Complexa, ressaltando seus aspectos geométricos, topológicos e analíticos. Começando com materiais preliminares, como resultados básicos sobre funções holomorfas de uma ou mais variáveis e a definição e primeiros exemplos de variedades complexas, passamos a uma introdução à teoria de feixes e sua cohomologia, ferramenta indispensável para o restante do trabalho. Após um estudo sobre fibrados de linha e divisores damos atenção à Geometria de Kähler e alguns de seus resultados centrais, como por exemplo o Teorema da Decomposição de Hodge, o Teorema ``Difícil\'\' e o Teorema das $(1,1)$-classes de Lefschetz. Em seguida, nos dedicamos ao estudo dos fibrados vetoriais complexos e sua geometria, abordando os conceitos de conexões, curvatura e Classes de Chern. Terminamos o trabalho descrevendo alguns aspectos da topologia de variedades complexas, como o Teorema dos Hiperplanos de Lefschetz e algumas de suas consequências. / The main goal of this work is to present a detailed study of the foundations of Complex Geometry, highlighting its geometric, topological and analytical aspects. Beginning with a preliminary material, such as the basic results on holomorphic functions in one or more variables and the definition and first examples of a complex manifold, we move on to an introduction to sheaf theory and its cohomology, an essential tool to the rest of the work. After a discussion on divisors and line bundles we turn attention to Kähler Geometry and its central results, such as the Hodge Decomposition Theorem, the Hard Lefschetz Theorem and the Lefschetz Theorem on $(1,1)$-classes. After that, we study complex vector bundles and its geometry, focusing on the concepts of connections, curvature and Chern classes. Finally, we finish by describing some aspects of the topology of complex manifolds, such as the Lefschetz Hyperplane Theorem and some of its consequences.

My primary argument in this thesis is that creation theologies significantly influenced early developments in the doctrine of the Trinity, especially in Augustine of Hippo’s theology. Thus this is a work of historical theology, but I conclude with proposals for how Augustine’s theologies of creation and the Trinity can be read fruitfully with modern theology. I critically analyse developments in trinitarian theologies in light of ideas that were held about creation. These include the doctrine of creation ‘out of nothing’ and ideas about other creative acts (e.g., forming or fashioning things). Irenaeus and other early theologians posited roles for God (the Father), the Word / Son, the Spirit, or Wisdom in creative acts without working out formal views on economic trinitarian acts. During the fourth century trinitarian controversies, creation ‘out of nothing’ and ideas about ‘modes of origin’ influenced thinking on consubstantiality and relations within the Trinity. Basil of Caesarea and others also presented ideas about trinitarian acts of creation and the Trinity in hexaemeral works. I will argue that in Augustine’s views of trinitarian acts of creation, he attributes roles to God (the Father), the Word / Son, and the Spirit. In his mature theology, he attributes the giving of formless existence, differentiated existence, and perfected existence to the three Persons respectively, while depicting shared roles. He also attributes to the Spirit the giving of the capability of ‘dynamic abiding’ to creatures, which gives them agency in continuing their existence. Augustine’s theologies of creation and the Trinity were significantly influenced by his exegesis of Gen. 1, John 1. 1-3, Wisdom, and other scriptures, and his ideas resonate with the hexaemeral works of Basil and Philo of Alexandria. I argue that scholars should examine these sources and Augustine’s own hexaemeral commentaries to gain a deeper understanding of his trinitarian theology.

The aim of this diploma thesis is to present one of the aspekts of teaching of early reading and writing in primary schools in England. By this aspekt we mean methods that are used during teaching early reading and writing. The theoretical part of the diploma thesis familiarize the reader with the education system in England as whole, describes the National Curriculum and closely familiarize the reader with the pre-school curriculum and primary school curriculum. From both curriculums are pointed out especially those aspects which relate to teaching English language. In addition, there will be presented programs of teaching early reading and writing and closely described methods of teaching early reading and writing. The practical part of diploma thesis is based on research which was performed in community primary school "Dale Community Primary School". The research was conducted by several methods: questionnaire survey, observation and controlled observation. The questionnaires and observation aimed at finding methods of teaching early reading and writing which are used at the specific primary school. In addition, two modelled lessons were prepared and later reflected. During both modelled lessons were used methods of teaching early reading and writing. The knowledge and information gained from...

The diploma thesis deals with detailed look at the project Education 21 and so the teaching using new technologies, especially interactive whiteboards and netbooks, in Czech schools. The objective was observation of teaching mathematics, methods of using interactive whiteboards and netbooks in mathematics lessons at secondary schools participating project Education 21. Evaluation and conclusions are based on analyzes of lessons recorded in the academic year 2009/2010 and 2010/2011, which were investigated and further divided into shorter stories in case that interactive whiteboard or netbook was used in some way. I watched purpose and the way of use of interactive whiteboard or netbook in the episodes, then the material which this technology was used with the aid of and who worked with interactive whiteboard. Some episodes can serve as an example of "good" or interesting way of working with the IWB and NTB in mathematics lessons, or they can be an inspiration for other mathematics teachers but also teachers of other subjects. Keywords: interactive whiteboard (IWB), netbook (NTB), interactive textbook - e- books, applet, mathematics, Education 21, smart notebook, videos.

Syftet med denna uppsats är att utforska betydelsen av Logos-begreppet hos två nytestamentliga författare, och sedan ta ställning till om det finns någon koppling mellan texternas användning av begreppet. Vid jämförelsen av Logos-begreppet kommer jag fram till att de båda författarna använder sig av Logos, dels på liknande sätt, men att de skiljer sig på vissa saker. Det skiljer sig även inom Johannesevangeliet, där det sker en förflyttning från att tro på det inkarnerade Logos till att höra Jesu talade Logos, som en premiss för att uppnå frälsning/räddning. Min slutsats om Jakobsbrevet är att åhörarna aktivt ska både höra och göra Logos, vilket liknar förekomsten av Logos-begreppet i Joh 4, 5 och 8 dock inte i Joh 1.

The knowledge of geometry of molecular complexes formed via molecular association in solution through weak interactions is always important to understand the origin of stability and function of an array of molecules, supramolecular assemblies, and macromolecular networks. Simple 1:1 molecular complexes are very useful in this regard as they provide a model to understand both the nature of these interactions and their structural implications. Several weak noncovalent forces from long range (van der Waal’s, electrostatic, induction, dispersion) to short range (charge transfer) govern the geometry, that is, relative orientation of the two molecules in such a complex. On one hand, we find 1:1 electron donor acceptor (EDA) complexes such as naphthalene-tetracyanobenzene, hexamethylbenzene-chloranil etc. which stack parallel or in slipped parallel geometry in their crystals. On the other, benzene dimer has been found to stabilize in T shaped geometry in all its three physical states. In this thesis, I focus on 1:1 EDA complexes in solution. A good volume of literature is available which deals with the optical studies on the formation of such complexes. It has been suggested that the nature of the intermolecular interactions stabilizing these complexes in the gas phase or in their crystals is modified by the presence of solvent-solute interactions in solution thus bringing in difference in the solution geometry. However, the existing experimental techniques, both optical and magnetic, are unable to determine the exact geometries of 1:1 EDA complexes in solution. This opens an opportunity to probe their geometry in solution. The quadratic nonlinearity or first hyperpolarizability (β) of a molecule is a measure of the change in dipole moment (or polarization) in the second order of the applied electrical field and thus has a purely electronic origin. It is a tensorial property and can be resolved in components along the three dimensions. The number of β components and the nonlinear optical anisotropies in a typical donor-acceptor type dipolar molecule, defined as (equation) (where1, 2, 3 axes define the molecular frame, 1 being the direction along the principal axis of symmetry and pointing from the acceptor toward the donor), are determined by the symmetry /structure of the molecule. It has been shown theoretically that the 1:1 EDA complexes possess large hyperpolarizabilities. In the case of pNA dimers calculation revealed that the geometry of the dimer and its symmetry is important for obtaining the correct estimate of β from its tensorial components. Therefore, it should be possible to use the values of tensorial β components to construct the unknown geometry of such complexes. Experimentally macroscopic depolarization ratios (D and D′) in the laboratory fixed frame (XYZ, X being the direction of polarization and Z the direction of propagation of the incident light), are measured from the polarization resolved intensities of second harmonic scattering from molecules in solution using the hyper-Rayleigh scattering technique. The depolarization ratios are correlated to the anisotropy parameters, u and v through a co-ordinate transformation. In this thesis I, have first, characterized the quadratic nonlinear optical property of a variety of 1:1 electron donor acceptor complexes and used the values of u and v obtained from depolarized hyper-Rayleigh scattering to deduce their geometry in solution. Chapter 1 provides an introduction to the 1:1 electron donor acceptor complexes, their relevance to chemistry and biology. It also contains an introduction to nonlinear optical processes in molecules. The objective of the present work and scope of the investigation carried out in this thesis is presented in this chapter. Chapter 2 describes the details of the experimental polarization resolved HRS technique. The geometrical model adopted for the analysis of the HRS data has also been introduced and the method of analysis has been described in detail in this chapter. Chapter 3 presents the measurement of β values of two series of 1:1 EDA complexes of variously substituted methylbenzenes donors with tetrachloro-p-benzoquinone (CHL) and dicyanodichloro-p-benzoquinone (DDQ) acceptors at 1064 nm. In agreement with recent theoretical results we find large first hyperpolarizabilities for these complexes. The β values are greater than that of the typical push-pull molecule p-nitroaniline (pNA). We also find that in general β decreases with decrease in the donor strength. Chapter 4 presents the β values for the two series of EDA complexes of CHL and DDQ acceptors at 1907 nm. The values of β are less in magnitude at 1907 nm than that at 1064 nm which is due to the dispersion effect in β. In Chapter 5 and 6, it is described how depolarized hyper-Rayleigh scattering can be utilized to probe geometries of 1:1 complexes in solution. Chapter 5 concentrates mainly on 1:1 EDA complexes of CHL and DDQ and TCNB (tetracyanobenzene), while chapter 6 contains examples of other 1:1 molecular complexes where the noncovalent interactions are much weaker, such as in benzene-naphthalene, benzene-methoxybenzene, benzene-hexafluorobenzene and benzene-chlorobenzene pairs. We find the geometry of 1:1 EDA complexes in solution in terms of tilt angle (θ) and twist angle (ϕ) between the donor and acceptor pairs. The angle θ varies from 29°-47° for different pairs of EDA complexes, while ϕ varies within 34° and 38°. We find that the geometry of 1:1 EDA complexes in solution is different (twisted and tilted cofacial and twisted ‘V’) from those in the crystalline or gaseous states (cofacial), if known. We find that both benzene-naphthalene and benzene-chlorobenzene pairs assume twisted ‘T’ shape geometry with θ = 82° and 85°, respectively, and φ = 38°, while benzene-hexafluorobenzene assumes a twisted ‘V’ shape. A strong solvent effect is seen in the geometry of the benzene- methoxybenzene complex. The tilt angle is 55° when chloroform is used as a solvent and it is 82° without chloroform. Chapter 7 is the concluding chapter where the main work done in this thesis is summarized and future directions are presented.

This study has attempted to show the overarching significance of the conquering motif in relation to discourse dynamics of the entire book of Revelation and the significance of salvific history for its syntagmatic understanding. Based on language-in-use as a whole between the model author and the model audience, syntagmatic analysis (i.e., SVU analysis) and associative analysis (i.e., sign-intertextual reading) are eclectically and concertedly utilized by means of sampling analysis. Utilizing this integrative method, the findings are as follows: (1) the interwoven network of the prologue (Rev 1:1-8) programmatically provides the paradigmatic reading strategy for understanding the key paraenetic motif in the rest of the book against the background of salvific history; (2) by summarizing the churches’ earthly prophetic roles – withdrawal and witness through martyrdom – in terms of conquering, the model author alerts his audience to the military significance of their daily actions or choices in their ordinary earthly lives through visionary communication; (3) just as the prologue preliminarily guides, the ever-forward-moving historical framework serves as an incentive device for the paraenetic-imperative in Rev 2-3 and 4-22.

The aim of this study was to, from a special educational approach, describe and analyse nine teachers and headmasters experiences and perceptions of the use of laptos, ”1-1”, as a tool for working with 13-16 year old pupils born outside of Sweden and with special educational needs (SEN). Interviews were used for data collection. The interview material was analysed with the phenomenografic analysis method and with meaning condensation method. The result was analysed through three perspectives on special education and the concept of inclusion as the study’s frame of reference. The major result showed that teachers perceived 1-1 facilitated adaption of instruction to multicultural instruction groups. 1-1 also enabled a more individualised and inclusive instruction when pupils could get indvidualised tasks and study material and a lot of technical support in their computers. Cooperation with schoolmates and with families was not facilitated with 1-1, rather were many difficulties exposed in this area.