Ingeniero Civil / El presente trabajo tiene como objetivo determinar el desempeño sísmico de marcos de momento con conexión viga-columna doble t soldadas. Para ello, se implementa la metodología presentada en FEMA p695, la cual permite cuantificar el desempeño sísmico de nuevos sistemas estructurales. Para implementar la metodología, se requiere determinar de forma explícita las curvas de carga vs deformación de todos los elementos estructurales que componen el sistema sismo-resistente. Para las vigas y columnas se emplean las curvas presentadas en FEMA-356, mientras que para determinar la curva de la conexión viga-columna, se utiliza el modelo de rigidez desarrollado por Swanson, 2000. Los resultados indican que un marco de momento con conexiones doble-t soldadas solo puede considerarse como marcos especiales (SMF) si el tamaño de sus vigas es de perfiles IN70 o inferior. Para marcos que requieran vigas de mayor dimensión, se deben considerar como marcos ordinarios (OMF), con el factor de reducción menor correspondiente. Este resultado ratifica lo expuesto en FEMA-350, donde se precalifica la conexión t-stub plegada para ser usada como SMF solo hasta perfiles tamaño W24.

This thesis evaluates various techniques for intra-set clustering of social media data from an industry perspective. The research goal was to establish methods for reducing the amount of redundant information an end user must review from a standard social media search. The research evaluated both clustering algorithms and string similarity measures for their effectiveness in clustering a selection of real-world topic and location-based social media searches. In addition, the algorithms and similarity measures were tested in scenarios based on industry constraints such as rate limits. The results were evaluated using several practical measures to determine which techniques were effective. / Graduate

La leucémie aiguë lymphoblastique (LAL-T) est une hémopathie maligne qui représente 10 à 15% des LAL pédiatriques et 25% des LAL de l’adulte. Bien que la prise en charge et le pronostic (rémission dans 80-85% des cas) des LAL se soient améliorés au cours des 10 dernières années en partie dû à une meilleure stratification thérapeutique de ces entités malignes, le tableau clinique et le devenir des patients atteints de LAL-T restent péjoratif avec environ 30% de rechute dans les 2 années qui suivent le diagnostic. Au cours de ces dernières années, des sous-types spécifiques de LAL-T associés à une valeur pronostique ont été décrits et des thérapies ciblées devraient pouvoir être proposées à l’avenir. Dans ce contexte, mon travail de thèse a permis de définir et de mieux comprendre les différents niveaux de dérégulation de MYC dans les LAL-T à travers l’analyse moléculaire et biochimique de MYC et de ses principaux régulateurs sur une large cohorte protocolaire de LAL-T pédiatriques et adultes. Tout d’abord, nous montrons que l'expression de MYC est très variable et que des niveaux d'expression élevés sont observés dans de nombreux cas en absence de mutations NOTCH1/FBXW7. De plus, nos travaux mettent en évidence que la dérégulation post-traductionnelle de MYC, via l'axe PI3K/AKT à travers l'inactivation de PTEN, constitue une voie majeure d'activation de MYC dans les LAL-T. Ainsi, l'ensemble de ces résultats confirment la pertinence d’envisager des stratégies thérapeutiques ciblant MYC pour le traitement des LAL-T. Mon projet de thèse a également consisté en la génération d’un modèle murin original permettant de suivre les clones tumoraux surexprimant Myc depuis les étapes de développement (pré-)tumoral les plus précoces jusqu’aux étapes finales de progression maligne. / T-cell Acute Lymphoblastic Leukemia (T-ALL) are malignant proliferations of thymocytes, which represent 10-15% of pediatric and 25% of adult ALL. Despite indisputable therapeutic progress, T-ALLs remain of poor prognosis. Patients often present with a high tumor load accompanied by a rapid disease progression, and about 30% of cases relapse within the first 2 years following diagnosis. It is now clear that significant improvements in therapy will require a more accurate knowledge of the oncogenes involved, as well as their oncogenic role within complex functional networks. In this context, my PhD project was focused on the understanding of the regulation of MYC in T-ALL. We demonstrate that MYC expression is highly variable and that high MYC expression levels can be generated independently of NOTCH1 pathway. Furthermore, we show that posttranscriptional deregulation of MYC constitutes a major alternative pathway of MYC activation in T-ALL, operating partly via the PI3K/AKT axis through down regulation of PTEN. Altogether, our results lend further support to the significance of therapeutic targeting of MYC in T-ALL pathogenesis. The second part of my project was to generate an original transgenic mouse model designed to “track” inducible MYC+ clones from the earliest steps of (pre-)malignant development to the onset of leukemia.

abstract: There are many biological questions that require single-cell analysis of gene sequences, including analysis of clonally distributed dimeric immunoreceptors on lymphocytes (T cells and B cells) and/or the accumulation of driver/accessory mutations in polyclonal tumors. Lysis of bulk cell populations results in mixing of gene sequences, making it impossible to know which pairs of gene sequences originated from any particular cell and obfuscating analysis of rare sequences within large populations. Although current single-cell sorting technologies can be used to address some of these questions, such approaches are expensive, require specialized equipment, and lack the necessary high-throughput capacity for comprehensive analysis. Water-in-oil emulsion approaches for single cell sorting have been developed but droplet-based single-cell lysis and analysis have proven inefficient and yield high rates of false pairings. Ideally, molecular approaches for linking gene sequences from individual cells could be coupled with next-generation high-throughput sequencing to overcome these obstacles, but conventional approaches for linking gene sequences, such as by transfection with bridging oligonucleotides, result in activation of cellular nucleases that destroy the template, precluding this strategy. Recent advances in the synthesis and fabrication of modular deoxyribonucleic acid (DNA) origami nanostructures have resulted in new possibilities for addressing many current and long-standing scientific and technical challenges in biology and medicine. One exciting application of DNA nanotechnology is the intracellular capture, barcode linkage, and subsequent sequence analysis of multiple messenger RNA (mRNA) targets from individual cells within heterogeneous cell populations. DNA nanostructures can be transfected into individual cells to capture and protect mRNA for specific expressed genes, and incorporation of origami-specific bowtie-barcodes into the origami nanostructure facilitates pairing and analysis of mRNA from individual cells by high-throughput next-generation sequencing. This approach is highly modular and can be adapted to virtually any two (and possibly more) gene target sequences, and therefore has a wide range of potential applications for analysis of diverse cell populations such as understanding the relationship between different immune cell populations, development of novel immunotherapeutic antibodies, or improving the diagnosis or treatment for a wide variety of cancers. / Dissertation/Thesis / Doctoral Dissertation Microbiology 2017

Humoral immunity provides protection against subsequent infections. Antigen-specific, high-affinity, class-switched antibodies are produced by B cells through rounds of proliferation, B cell receptor rearrangement and selection in the germinal centres (GC). T cells play an essential and indispensable role in this process and in the recent years the term T follicular helper cells (TFH) was coined to describe this cell subset. The aim of my thesis is to investigate whether there is more than one type of T cells within the TFH population and whether it has important functional consequences. Firstly, I use sheep red blood cell immunisation (SRBC) and Salmonella enterica infection to show phenotypical differences between TFH expressing high and low level of surface molecule PD-1. In order to investigate the relationship between different TFH populations gene profiling was carried out on the microarray platform. Detailed transcriptome analysis revealed the discrete nature of isolated TFH cell subsets and provided an overview of their genetic landscape. Secondly, I have investigated the dependence of TFH subsets on cognate interactions with B cell in SRBC model by generating BM chimeras. I have demonstrated that generation of PD-1HI TFH, but not of PD-1LO TFH, depends on antigen presentation by B cells. Furthermore, I have shown that provision of wild-type but not MHC II knock-out B cells rescues PD-1HI formation in BM chimeras after SRBC immunisation. Finally, I have explored plasticity within TFH subsets and showed that none of the populations is in a terminally differentiated state, as they can convert into one another. Thirdly, experiments with S. enterica model revealed that the absence of PD- 1HI TFH is independent of the splenic architecture disruption present within the first week of the response. Surprisingly, co-immunisation studies showed that PD-1HI population is not only present but even enhanced in the group which received both SRBC and S. enterica when compared to single immunisations. The work presented in the thesis documents that there is a significant and previously unappreciated heterogeneity within TFH subset. This knowledge is important for designing optimal vaccine strategies and treating autoimmune diseases, as in both processes the antibody production plays a crucial role and its manipulation (either enhancing or blocking antibody production, respectively) can significantly improve clinical interventions.

Previous research has provided evidence that CD8 T cells mediate immunity against infection with Theileria parva. However, the immunity induced by one parasite strain doesn‟t give complete protection against other strains and this is associated with parasite strain specificity of the CD8 T cell responses. There is evidence that such strain specificity is a consequence of the CD8 T cell responses of individual animals being focused on a limited number of immunodominant polymorphic peptide-MHC determinants. Dominant responses to the Tp2 antigen have been demonstrated in animals homozygous for the A10 MHC haplotype. Three Tp2 epitopes recognised by A10+ animals (Tp249-59, Tp250-59 and Tp298-106) have been defined. This project set out to investigate the dominance of these epitopes and to examine the T cell receptor (TCR) repertoires of the responding T cells. The specific objectives were to: (i) Determine the dominance hierarchies of the three defined Tp2 epitopes in both A10-homozygous and -heterozygous cattle. (ii) Examine the clonal repertoires of epitope-specific responses by analysis of TCR gene expression. (iii) Isolate full-length cDNAs encoding TCR α and β chain pairs from T cell clones of defined epitope specificity and use them to generate cells expressing the functional TCRs. Using MHC class I tetramers the relative dominance of CD8 T cell responses were found to differ between A10-homozygous and heterozygous cattle. All A10-homozygous cattle examined had detectable responses to all 3 Tp2 epitopes, the Tp249-59 epitope consistently being the most dominant. By contrast, only some A10-heterozygous cattle had detectable responses to Tp2 and when present the response was specific only for the Tp298-106 epitope. Analyses of the sequences of expressed TCR β chains showed that the responses in individual animals were clonotypically diverse, but often contained a few large expanded clonotypes. The TCRs of Tp298-106–specific T cells showed preferential usage of the Vβ13.5 gene and the frequent presence of a “LGG” motif within the CDR3 of the B chain. A conserved (public) TCRβ clonotype shared by the Tp250-59-specific CD8 T cells from all A10-homozygous cattle was identified. The TCRα chains co-expressed with this public TCRβ clonotype were identified for a number of T cell clones. Lentivirus transduction of Jurkat cells with three full-length TCR α and β chain pairs resulted in successful expression of one of the α/β chain pairs as a functional TCR, thus providing the basis for future work to generate bovine T cells expressing defined TCRs in vitro.

A criação e atualização de bases de conhecimentos com dados que comprovem, ou não, a eficácia de métodos, técnicas ou produtos de software podem ser facilitadas por meio de estudos experimentais e suas replicações em diferentes contextos, para que a base de conhecimento possa ser ampliada e consolidada. Porém, a condução dessas replicações gera uma grande quantidade de dados que devem ser coletados e armazenados, e posteriormente analisados, sendo que a gestão dessas informações de forma manual é propensa a erros. Nesse contexto, o Projeto Readers - um projeto de pesquisa colaborativa formado por pesquisadores brasileiros e norte-americanos - no qual este trabalho está inserido, teve como objetivos principais desenvolver, validar, empacotar e consolidar uma base de conhecimento sobre algumas técnicas, como por exemplo a PBR (Perspective Based Reading), que é uma técnica de leitura para detecção de defeitos em artefatos de software. Com as várias replicações de estudos experimentais que foram conduzidas no âmbito desse projeto, em particular replicações do Experimento PBR (Basili et al., 1996), ficou evidenciada a necessidade de apoio computacional em diversas etapas do processo de experimentação. Sendo assim, neste trabalho foi definida, modelada e implementada a ferramenta ExpVVT com a finalidade de automatizar as atividades de Planejamento e Operação do Experimento PBR. A ferramenta foi implementada com base em uma estrutura de meta-tabelas, o que facilita possíveis alterações no âmbito desse experimento. Salienta-se que o Pacote de Laboratório, o qual contém todos os artefatos necessários para que um estudo experimental possa ser replicado, também fica armazenado eletronicamente, com o apoio da ferramenta, o que facilita o registro de sua evolução. O uso da ferramenta foi explorado simulando-a com os dados de uma das replicações do Experimento PBR que foi conduzida no contexto do Projeto Readers. Embora tenha sido apenas uma simulação, pode-se perceber o auxílio que essa ferramenta pode trazer para futuras replicações desse experimento, reduzindo os custos associados com todo material necessário para uma replicação e também guiando os participantes do experimento, para que as atividades sejam feitas de forma mais controlada. / The creation and update of knowledge bases with data that provide evidences of the efficacy, or non-efficacy, of software methods, techniques or products can be facilitated by experimental studies and their replications in diverse contexts, so that the knowledge base can grow and be consolidated. However, replications generate great amount of data which must be collected, stored, and on later, analyzed, making the non-automated management of such information very error-prone. In this context, the Readers Project ? a collaborative research project formed by Brazilian and North-American researchers ? which this work is part of, has had as main goals the development, validation, packaging and consolidation of a knowledge base on some techniques, such as PBR (Perspective Based Reading), which is a reading technique used to defect detection in software artifacts. With the several replications of experimental studies that were conducted along this project, in particular the replications of the PBR Experiment (Basili et al., 1996), it has become evident the need of computational support in several activities of the experimentation process. Therefore, in this work, the ExpVVT tool was defined, designed and implemented with the goal to automate the Planning and Operation activities of the PBR Experiment. The tool was implemented based on a meta-table structure, which facilitates possible evolutions in this experiment context. It is emphasized that the Lab Package, which contains all the necessary artifacts needed to replicate an experimental study, is also stored electronically, with the tool support, which facilitates the tracing of its evolution. The tool?s use was explored simulating with data of some of the PBR Experiment replications which were conducted at the Readers Project context. Although just a single simulation took place, it can already be noticed the support that the tool can provide to future replications of this experiment, reducing the costs associated with every necessary material in a replication and also guiding the experiment participants, so that the activities are made in a more controlled way.

Les ailes d’avion sont composées d’une part croissante de pièces composites. Afin de réduire les coûts de production et de certification des assemblages, une nouvelle architecture en caissons creux, en fabrication ‘one shot’, est proposée. Cette structure contient des liaisons en T entre des sections sandwichs. Le comportement et les modes de dégradation de cette structure sont méconnus. Une méthodologie multi-échelle est mise en place pour appréhender l’endommagement de la voilure et ainsi améliorer l’outil numérique de dimensionnement.Tout d’abord, des systèmes essai – éprouvette spécifiques sont conçus pour représenter les modes de sollicitation de la liaison dans la voilure. Une analyse fine des essais instrumentés permet d’appréhender les scénarios d’endommagement pour chaque mode. Les premiers stades sont principalement pilotés par le nid d’abeille (cisaillement, déchirement). Un modèle numérique 3D permet de représenter les premiers stades d’endommagement. Il est construit avec une représentation en coque du nid d’abeille intégrant les variabilités de la structure (collage, déformation de cellules). Egalement, une loi de comportement élasto-plastique endommageable des matériaux constitutifs de la voilure (plis unidirectionnels et tissés) est développée et implémentée pour les stratifiés multi-matériaux. Sur les bases de ce modèle, une modélisation simplifiée du comportement de la liaison est développée. Elle consiste à assembler des coques par des connecteurs avec des propriétés élasto-plastiques. La création d’un calcul prédictif de voilure intègre le modèle simplifié de comportement de la liaison. / Aircraft wings are composed of a growing composite part portion. To reduce production and assembly certification costs, a new hollow box architecture is proposed with a one shot manufacturing process. This structure includes T-connections between honeycomb sandwich panels. Mechanical behavior and degradation modes are unknown for this structure. A multiscale methodology is developed to deepen the wing damage knowledge and to improve the numerical sizing tool.Firstly, specimen-testing couples are designed to represent the loading modes of the T-connections. A detailed analysis of instrumented testing allows to understand the damage scenarios for each mode. The first damage stages are mainly driven by the honeycomb behavior (shear, tear). A 3D numerical model represents the first damage stages. It is built with a shell representation of the honeycomb which integrates the structure variability (bonding, deformation of cells). An elastic-plastic damage law (unidirectional and woven plies) is developed for the wing materials. Then, it is implemented for multi material laminate. Based on this model, a simplified modeling is realized for the connection behavior. It consists of assembling shells by connectors with elastic-plastic properties. Building a predictive wing simulation requires a simplified model integration of the T-bond behavior.

Abstract Osteoarthritis (OA) is a common chronic condition that reduces the quality of life of millions of individuals worldwilde. In OA, the progressive degradation of articular cartilage (AC) and bone can cause joint pain and disability. Currently there is no effective treatment available for OA besides gross joint replacement surgery. This is largely due to the lack of accurate biomarkers to test potential drugs and treatments that could stop or reverse the disease progression. The primary tools in use for OA diagnostics are inadequate in the early stage of the disease and merely allow its late manifestations to be visualized. Quantitative MRI (qMRI) has demonstrated the ability to distinguish degenerated from intact AC prior to radiographic changes. Moreover, so-colled rotating frame relaxation time parameters are sensitive to the slow molecular motion domain, relevant for clinical applications. T1ρ and T2ρ relaxation time measurements using adiabatic spin-lock pulses (AdT1ρ and AdT2ρ) have shown superior sensitivity to cartilage degeneration in vitro over conventional qMRI parameters. This thesis aimed to compare the differences that exist between established qMRI methods for AC against arthroscopic evaluation as well as optimize and validate AdT1ρ and AdT2ρ mapping of cartilage in vivo. The findings showed that T1 and T2 relaxation time and delayed gadolinium enhanced MRI of cartilage (dGEMRIC) were able to discriminate among different arthroscopic grades of cartilage lesions. However, arthroscopic findings and qMRI parameters were not correlated, supporting the viewpoint that qMRI may be able to elicit more comprehensive information on the quality of remaining cartilage tissue as compared to diagnostic arthroscopy, which detects cartilage loss. AdT1ρ and AdT2ρ mapping of cartilage in vivo were successfully implemented and validated on a clinical MRI system. AdT1ρ and AdT2ρ were able to overcome limitations of the existing methods (continuous wave spin-lock approach), which are challenging to implement on clinical settings for their susceptibility to field inhomogeneity and relatively high power deposition in tissue. The association of AdT1ρ and AdT2ρ with cartilage and bone marrow lesions and osteophytes was demonstrated. The findings indicate the utility of AdT1ρ and AdT2ρ mapping as potential biomarkers for evaluation of early cartilage degeneration in OA research and clinical applications. / Tiivistelmä Nivelrikko on tavallinen krooninen sairaus, joka huonontaa miljoonien ihmisten elämänlaatua ympäri maailman. Nivelrikossa nivelruston ja luun asteittain etenevä rappeuma voi aiheuttaa nivelkipua ja liikuntakyvyttömyyttä. Tällä hetkellä nivelrikkoon ei ole muuta tehokasta hoitokeinoa kuin tekonivelleikkaus. Tämä johtuu suuremmaksi osaksi tarkkojen biomarkkereiden puutteesta, joiden avulla voitaisiin kokeilla mahdollisia nivelrikon pysäyttäviä tai tautia parantavia lääkkeitä tai hoitokeinoja. Nykyisin nivelrikon diagnostiikassa käytetyimmät menetelmät ovat riittämättömiä sairauden alkuvaiheessa, ja ne tunnistavat vain sairauden loppuvaiheet. Kvantitatiivisilla magneettikuvausmenetelmillä pystytään erottamaan rappeutunut ja ehjä nivelrusto toisistaan ennen röntgenkuvissa havaittavia muutoksia. Nk. pyörivän koordinaatiston relaksaatioaikaparametrit ovat herkkiä havaitsemaan hidasta molekyyliliikettä, jolla on merkitystä kliinisissä sovelluksissa. Kokeellisissa olosuhteissa T1ρ- ja T2ρ-relaksaatioaikojen mittaaminen käyttäen adiabaattisia spin-lukkopulsseja (AdT1ρ ja AdT2ρ) on osoittautunut erityisen tarkaksi nivelruston rappeutumisen toteamiseksi verrattuna perinteisiin kvantitatiivisiin magneettikuvausmenetelmiin. Tämän väitöskirjan tavoitteena oli vertailla vakiintuneita nivelrikon kvantitatiivisia magneettikuvausmenetelmiä sekä optimoida ja validoida AdT1ρ- ja AdT2ρ-menetelmien käyttöä ihmisen ruston tutkimisessa. Tulokset osoittivat, että T1- ja T2-relaksaatioajat sekä varjoainetehosteinen nivelruston magneettikuvaus pystyivät erottelemaan niveltähystyksessä määritettyjä nivelrikon eri asteita. Kuitenkaan niveltähystyslöydökset ja kvantitatiiviset magneettikuvausparametrit eivät korreloineet viitaten kvantitatiivisen magneettikuvauksen kykyyn osoittaa tarkemmin jäljellä olevan ruston laatua verrattuna niveltähystykseen, jossa arvioidaan ruston puutosta. Nivelruston AdT1ρ- ja AdT2ρ-relaksaatioaikamittaukset onnistuttiin suorittamaan ja validoimaan vapaaehtoisilla koehenkilöillä kliinisellä magneettikuvauslaitteella. AdT1ρ ja AdT2ρ eivät kärsineet samoista puutteista kuin tavanomaiset jatkuva-aaltoiset spin-lukkomenetelmät, joka ovat herkkiä kenttäepähomogeenisuuksille ja aiheuttavat suuremman radiotaajuusaltistuksen. Tutkimuksessa osoitettiin AdT1ρ- ja AdT2ρ-mittausten ja luuödeemin sekä osteofyyttien välinen yhteys. Tulokset osoittivat, että AdT1ρ- ja AdT2ρ-mittaukset ovat potentiaalisia biomarkkereita varhaisen nivelruston rappeuman havaitsemiseksi tutkimus- ja kliinisessä käytössä.

M.A. (African Languages) / The aim of this study is to endeavour to analyze the language and style in W.T. Matlala's Molato mpeng and Montshepetsa bosego. The study will focus on how the writer uses language, how he assumed a particular manner of narrating by meticulously using a mixture of his dialect and standard Northern Sotho. This is done in the introductory chapter. In the second chapter of our study we tried to develop a stylistic framework whose purpose is to inform the analysis of Matlala's two novels. In the third chapter of this study, an analysis of Matlala's two prose fictions will be undertaken. The choice of words, sentences and paragraphs will be analyzed. The fourth chapter of this work, will attempt to show how the writer has used imagery, proverbs, idioms and dialogue as other stylistic features to promote understanding and to bring images of the mind to the life of the literary work. Chapter five is the conclusion - we will look at how Matlala has synthesized all the stylistic features in communicating his emotions, attitudes, thoughts as well as giving freshness and vigour to his works.