La Dystrophie Myotonique de type 1 (DM1) est cause par l’expansion anormale d’un triplet CTG dans la partie 3’UTR du gène DMPK, entrainant l’agrégation du transcrit mutant dans des inclusions ribonucléoprotéiques appelées foci. D’après plusieurs études structurales sur des courtes répétitions CUG, il a été proposé que les expansions CUG se replient en une structure en tige-boucle qui interfère avec l’activité de plusieurs facteurs lié au métabolisme de l’ARN et altère leur fonction cellulaire. Le facteur d’épissage muscleblind-like 1 (MBNL1) a été identifié par sa capacité à interagir avec les répétitions CUG. In vivo, ces répétitions entrainent la séquestration de cette protéine aboutissant en une déplétion nucléaire. Un autre facteur d’épissage, la CUG Binding protein (CUGBP1), est également impliqué dans la pathologie. Au lieu d’être séquestré par les répétitions, la stabilité protéique de CUGBP1 est augmentée dans les tissus DM1 entrainant un gain d’activité pour ce facteur. La séquestration de MBNL1 et la stabilisation de CUGBP1 résultent en la dérégulation de l’épissage alternatif de plusieurs transcrits musculaires et du cerveau et la réexpression d’isoformes protéiques fœtales dans les tissus adultes. Cependant, de récentes études suggèrent que d’autres facteurs ou voies de signalisation que celles faisant intervenir MBNL1 et CUGBP1 pourraient être impliquées dans la pathologie DM1.Le but de mon travail de thèse a été d’identifier de nouveaux facteurs ayant la capacité d’interagir avec les répétitions CUG. A l’aide d’une purification sur chromatographie d’affinité utilisant un ARN contenant 95 répétitions CUG comme appât, nous avons identifié l’ARN hélicase p68/DDX5. p68 fait partie de la famille des protéines DEAD-box, caractérisée par un core protéique conservé constitué de neufs domaines hautement conservés, dont le motif DEAD, à l’origine du nom de ces protéines. p68 est impliquée dans de nombreux aspects du métabolisme de l’ARN, dont la transcription, l’épissage, l’export, la traduction et la dégradation des ARN. Nous avons montré, que p68 colocalise avec les foci CUG dans un modèle cellulaire exprimant la partie 3’UTR du gène DMPK contenant de longues répétitions CTG. Nous avons identifié que p68 augmente l’interaction de MBNL1 sur les répétitions CUG et une structure secondaire particulière d’un élément régulateur de l’ARN pré-messager cardiac Troponin T (TNNT2), dont l’épissage est dérégulé dans la pathologie. L’insertion de mutations dans le core de l’hélicase de p68 abolit l’effet de p68 sur la fixation de MBNL1 ainsi que la colocalisation de p68 avec les expansions CUG in vivo, suggérant que le remodelage des structures secondaires ARN de manière ATP-dépendante par p68 facilite l’interaction de MBNL1. Nous trouvons également que la compétence de p68 pour réguler l’inclusion de l’exon alternatif 5 de TNNT2 dépend de l’intégrité des sites de fixation de MBNL1.Nous proposons que p68 agit comme un modificateur de l’activité de MBNL1 sur ces cibles d’épissage ainsi que sur les expansions CUG à l’origine de la pathologie. / Myotonic Dystrophy type I (DM1) is caused by an abnormal expansion of CTG triplets in the 3’ UTR of the DMPK gene, leading to the aggregation of the mutant transcript in nuclear RNA foci. Based on structural studies on short CUG repeats, it has been proposed that expanded CUG repeats fold into an imperfect hairpin structure that interferes with the activities of RNA binding proteins and alters their normal cellular function. The muscleblind-like 1 protein (MBNL1) was identified by its ability to bind to CUG repeats. It has been shown that the expanded mutant transcript promotes the sequestration of the MBNL1 splicing factor in nuclear RNA foci. CUGBP1 is another splicing factor that is involved in DM1. Instead of being sequestered by the repeats, the steady-state level of CUGBP1 is increased in DM1 tissues, leading to a gain of activity of the protein. The sequestration of MBNL1 and the up-regulation of CUGBP1 in DM1 results in the misregulation of alternative splicing of a subset of muscle and brain-specific transcripts, leading to the re-expression of fetal isoforms in adult tissues. However, several recent studies suggest that factors or signaling pathways other than MBNL1 and CUGBP1 could be involved in DM1 pathogenesis.The aim of this work was to isolate new factors that bind to CUG repeats. Using an affinity chromatography strategy with an RNA containing 95 pure CUG repeats, we identified the RNA helicase p68 (DDX5). p68 is a prototype of DEAD-box RNA helicase proteins. This family is characterized by a conserved core, consisting of nine conserved motifs including the DEAD signature, which gives rise to the name to these proteins. p68 is involved in many aspects of RNA metabolism including transcription, RNA processing, RNA export, translation and mRNA degradation. We showed that p68 colocalized with RNA foci in cells expressing the 3’UTR of the DMPK gene containing expanded CTG repeats. We found that p68 increased MBNL1 binding onto pathological repeats and the stem-loop structure regulatory element within the cardiac Troponin T (TNNT2) pre-mRNA, splicing of which is misregulated in DM1. Mutations in the helicase core of p68 prevented both the stimulatory effect of the protein on MBNL1 binding and the colocalization of p68 with CUG repeats, suggesting that remodeling of RNA secondary structure through a ATP-dependant manner by p68 facilitates MBNL1 binding. We also found that the competence of p68 for regulating TNNT2 exon 5 inclusion depended on the integrity of MBNL1 binding sites.We propose that p68 acts as a modifier of MBNL1 activity on splicing targets and pathogenic RNA.

Orientador: Luzia Aparecida Trinca / Resumo: Nas mais diversas áreas do conhecimento se procura aumentar a eficiência dos delineamentos experimentais, principalmente, para minimizar os custos das pesquisas. O uso dos delineamentos ótimos, com seus diferentes critérios de otimização, é fundamental para se obter resultados que maximizam a informação em estudos experimentais. A maioria dos métodos pressupõe homogeneidade de variâncias, a qual nem sempre é veri cada no conjunto de dados. O objetivo deste trabalho é desenvolver uma metodologia para construção de delineamentos ótimos exatos e cientes em situações de variância não homogênea. Assume-se que linearidade e homoscedasticidade são obtidas via o uso de transformações da família Box-Cox e, além de critérios de otimização puros, critérios compostos que combinam duas propriedades são propostos. Resultados para vários exemplos sob os modelos de primeira e segunda ordem são obtidos e discutidos. / Abstract: In several areas of knowledge we seek to increase the efficiency of experimental designs, mainly in order to minimize the costs of reaserch. The use of optimal design with different optimization criteria is fundamental to obtain results that maximize the information in experimental studies. Most of the methods assume homogeneity of variances, which is not always verified in the data set. The goal of this work is to develop a methodology to construct exact optimal or efficient designs in situations of nonhomogeneous variance. It is assumed that application of a transformation from the Box-Cox family accomplish both linearity and homocedasticity. Pure design criterion as well as compound criteria using two desired properties are used. Results for several examples assuming first and second order models are presented and discussed. / Doutor

Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2016-03-09T14:22:52Z No. of bitstreams: 1 PDF - Ingredy Meneses Cavalcanti de Albuquerque.pdf: 3429704 bytes, checksum: 2b4640e1e1ebd9b5bfdbca2369dc42ab (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2016-07-21T21:01:42Z (GMT) No. of bitstreams: 1 PDF - Ingredy Meneses Cavalcanti de Albuquerque.pdf: 3429704 bytes, checksum: 2b4640e1e1ebd9b5bfdbca2369dc42ab (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2016-07-21T21:01:49Z (GMT) No. of bitstreams: 1 PDF - Ingredy Meneses Cavalcanti de Albuquerque.pdf: 3429704 bytes, checksum: 2b4640e1e1ebd9b5bfdbca2369dc42ab (MD5) / Made available in DSpace on 2016-07-21T21:01:49Z (GMT). No. of bitstreams: 1 PDF - Ingredy Meneses Cavalcanti de Albuquerque.pdf: 3429704 bytes, checksum: 2b4640e1e1ebd9b5bfdbca2369dc42ab (MD5) Previous issue date: 2014-11-14 / Biodiesel is a fuel derived from biomass sources, among them vegetable oils from agroindustrial activities. As a promising fruit for use as bioenergy matrix in the biodiesel synthesis highlights the soursop (Annona muricata L.) due to its potential productivity in semi-arid region of Northeast Brazil, which is favored by its edaphoclimatic conditions. Thus, the aim of this work was to optimize the reaction conditions for transesterification via ethanolysis for biodiesel production from soursop seed oil. It was first performed an exploratory study using Plackett-Burman design with the following variables: reaction time, oil/alcohol molar ratio, catalyst concentration, and temperature. It was identified as statistically significant variables catalyst concentration, temperature and oil/alcohol molar ratio, which were employed in a Rotatable Central Composite Design. However, the model used to describe this process exhibited lack of fit. This drawback was overcome with the implementation of a BoxBehnken Design, whose greatest mass yield of 94.99% was obtained using 1.25% of catalyst concentration, temperature of 50 °C and oil/alcohol molar ratio of 1:10. The built model was used to establish an optimal experiment, at a 95% confidence level, with a 95.51% determination coefficient for the following experimental conditions: catalyst concentration of 1.18%, temperature of 52.20 °C, and oil/alcohol molar ratio of 1:8.39. Finally, the physical and chemical analysis showed the conditions for obtaining biodiesel from soursop seed oil as a promising alternative to its mixture with petroleum diesel. / O biodiesel é um combustível derivado de fontes de biomassa, dentre elas óleos vegetais provenientes de atividades agroindustriais. Dentre as promissoras fruteiras a serem utilizadas como matriz bioenergética na síntese do biodiesel destaca-se a graviola (Annona muricata L.), devido ao seu potencial produtivo no semiárido nordestino favorecido pelas características edafoclimáticas da região. Assim, objetivou-se com este trabalho otimizar as condições reacionais por transesterificação via rota etílica para produção de biodiesel do óleo de sementes de graviola. Inicialmente foi realizado um estudo exploratório utilizando planejamento Plackett-Burman com as variáveis: tempo reacional; razão molar óleo/álcool; porcentagem do catalisador; e temperatura. Foram identificadas como variáveis estatisticamente significativas porcentagem de catalisador, temperatura e razão molar óleo/álcool, as quais foram empregadas em um Planejamento Composto Central Rotacional. Todavia, o modelo utilizado para descrever o processo apresentou falta de ajuste. Este inconveniente foi superado com a execução do planejamento Box Behnken, cujo maior rendimento mássico de 94,99% foi obtido usando porcentagem de catalisador de 1,25%, temperatura de 50 ºC e razão molar óleo/álcool de 1:10. Com o modelo construído foi possível estabelecer como ótimo experimental, a um nível de confiança de 95%, com coeficiente de determinação de 95,51% para as seguintes condições experimentais: porcentagem de catalisador de 1,18%, temperatura de 52,20 ºC e razão molar óleo/álcool de 1:8,39. Por fim, as análises físicas e químicas apontaram para condições de obtenção do biodiesel de graviola como uma alternativa promissora para mistura em diesel de petróleo.

A caracterização estática dos sistemas caóticos clássicos dissipativos tem sido realizada através do cálculo das dimensões generalizadas \'D IND. q\' e do espectro de singularidades f(alfa). Os métodos mais comuns de cálculo numérico dessas funções utilizam algoritmos de contagem de caixa. Porém, esses algoritmos produzem um erro sistemático através de \'caixas espúrias\', levando a resultados distorcidos. Por essa razão, estudamos métodos numéricos que não utilizam o algoritmo de contagem de caixa, verificando em que casos eles podem ser aplicados eficazmente e propusemos um novo algoritmo de contagem de caixa que reduz o número de \'caixas espúrias\', obtendo melhores resultados. / The static caracterization of classical dissipative chaotical systems has been achieved by the calculation of the generalized dimensions \'D IND. q\' and the spectrum of singularities f(alfa). The most used numerical methods of evaluating these functions are based on box counting algorithms. The results obtained by those methods are distorced by the presence of \'spurious boxes\' generated intrinsecally by these algorithms. For this reason, we have studied numerical methods that don\'t use box counting algorithms, and we have tried to verify in which kind of sets they give best results. We also have proposed a new box counting algorithm that reduces the number of \'spurious boxes\', and led to better results.

A caracterização estática dos sistemas caóticos clássicos dissipativos tem sido realizada através do cálculo das dimensões generalizadas \'D IND. q\' e do espectro de singularidades f(alfa). Os métodos mais comuns de cálculo numérico dessas funções utilizam algoritmos de contagem de caixa. Porém, esses algoritmos produzem um erro sistemático através de \'caixas espúrias\', levando a resultados distorcidos. Por essa razão, estudamos métodos numéricos que não utilizam o algoritmo de contagem de caixa, verificando em que casos eles podem ser aplicados eficazmente e propusemos um novo algoritmo de contagem de caixa que reduz o número de \'caixas espúrias\', obtendo melhores resultados. / The static caracterization of classical dissipative chaotical systems has been achieved by the calculation of the generalized dimensions \'D IND. q\' and the spectrum of singularities f(alfa). The most used numerical methods of evaluating these functions are based on box counting algorithms. The results obtained by those methods are distorced by the presence of \'spurious boxes\' generated intrinsecally by these algorithms. For this reason, we have studied numerical methods that don\'t use box counting algorithms, and we have tried to verify in which kind of sets they give best results. We also have proposed a new box counting algorithm that reduces the number of \'spurious boxes\', and led to better results.

The purpose of this thesis work was to develop an automatic fault detection system for surveillance of bearing temperature in hydropower turbines. The parameters used except the bearing temperature were cooling water temperature and cooling water flow. A simple static model based on data sampled every minute was developed to estimate the bearing temperature. Then a detector for detection of change in bearing temperature based on the CUSUM-algorithm was designed. Since the amount of data was very small the developed model was too uncertain to be used in a working system. The designed fault detection system showed to work well for the available data. It is, however, recommended that the performance of the system should be evaluated using more data. Another model based on data sampled once every minute for at least a year has to be developed before the system can be fully evaluated. The results shown were: • The fault detection system can discover fast and slow changes in bearing temperature. • No false alarms were given for measuring faults and sensor faults of the types used in this thesis. If a measuring fault occurs for too long there will be an alarm. The fault detection algorithm was also implemented in Delphi to be used in a working system over the Internet where for example trends and alarms will be presented. / Syftet med examensarbetet var att utveckla ett automatiskt feldetekteringssystem för övervakning av bärlagertemperaturen i vattenkraftturbiner. De ingående parametrarna förutom bärlagertemperaturen var kylvattentemperaturen och kylvattenflödet. En enkel statisk modell baserad på data samplat en gång per minut togs fram för att estimera bärlagertemperaturen. Därefter utvecklades en detektor för att upptäcka avvikelser i bärlagertemperaturen baserad på CUSUM-algoritmen. På grund av en för liten mängd data var den framtagna modellen alltför osäker för att kunna implementeras i ett fungerande system. Det framtagna feldetekteringssystemet visade sig fungera bra för de data som fanns tillgängliga. Det är däremot rekommenderat att utvärdera systemets prestanda med längre dataserier. En ytterligare modell baserad på minutdata över ett år måste tas fram innan systemet kan fungera på riktigt. De resultat som erhölls var: • Feldetekteringssystemet klarar av att upptäcka abrupta och långsamma avvikelser av bärlagertemperaturen. • Inga falsklarm ges då det är enstaka mätfel eller givarfel av sådan typ som tagits upp i arbetet. Pågår ett mätfel alltför länge ges dock ett larm. Feldetekteringsalgoritmen implementerades även i Delphi för att kunna användas i ett fungerande system över Internet där t.ex. trendkurvor och larmsignaler skall kunna presenteras.

Despite its close proximity to downtown, East Austin is one of the underprivileged and under-developed areas in the City of Austin. Ethnic minorities and low-income persons in inner-city areas often lack access to big box retail due to these stores being disproportionately located outside of their neighborhoods. The aim of this study is to identify the current accessibility of big box retail for East Austin’s residents in order to confirm the potential impacts of big box retail growth on minority and lowincome populations. Using GIS-based network analysis, it is possible to measure whether the residents in East Austin have equal access to big box retail stores, as compare to other Austin areas. Although residents in East Austin have greater accessibility to other neighborhood-type retail like drug stores, small-format value stores, and supermarkets, they must travel farther to access community-type retail like home improvement stores,department stores, large-format value stores. Moreover, these populations have access to fewer cars, and must rely on public transit. Socio-economic characteristics of East Austin include a high percentage of individuals living below the poverty line, high disability rates, low to no vehicle ownership, and high percentages of female headed households. Finally, the study proposes new mixed-use, mixed-income development models as a way to improve retail access to minority and low-income population. / text

Baigiamajame darbe išanalizuota mokslinės literatūros šaltiniai, kuriuose buvo tirti ir išnagrinėti riedmenų ašidėžių temperatūrų kitimo procesai, temperatūrų pasiskirstymas ašidėžės ir aširačio mazgo elementuose. Išanalizuoti ašidėžių temperatūrų kitimo greičiai esant guolių gedimams, pateiktos ašidėžių liekamojo resurso prognozės. Išnagrinėti tyrimai pagrindžiantys riedmenų automatinės kontrolės prietaisų išdėstymo atstumų atitikimą saugių eismo sąlygų reikalavimams. Darbe pateikta realiomis sąlygomis atliktų bandymų rezultatų analizė. Nustatytos ašidėžių mazgo kaitimo tendencijos. Pateikta pagrindinių faktorių galinčių įtakoti ašidėžių kaitimo intensyvumą. Atliktas gautų bandymo rezultatų palyginimas si literatūros šaltiniuose pateiktais duomenimis ir nurodytos galimos bandymų rezultatų ir literatūros šaltinių duomenų neatitikimo priežastys. Pateiktos rekomendacijos, kuriomis remiantis galima padidinti traukinių eismo saugą ir išvengti avarinių situacijų dėl ašidėžių mazgo galimų defektų ir galima prognozuoti liekamąjį darbo resursą. Nurodytos tolesnių papildomų tyrimų kryptys. Baigiamąjį darbą sudaro 3 pagrindinės dalys, išvados, siūlymai bei literatūros sąrašas. Darbo apimtis – 74 p. teksto be priedų, 49 iliustr., 12 lentelių, 29 literatūros šaltiniai. / In the final analysis of the scientific literature sources that have been investigated and examine the axle box temperature variation processes, temperature axle box and of the wheelset node items. Parse axle box temperature variation speeds at bearing failures, the axle box residual resource forecast. The studies on which the automatic control devices layout distances with safe traffic conditions. Work in real terms of the results of the tests carried out. The axle box assembly heating trends. The main factors which could influence the axle box heating. The comparison of the results of the test carried out si literature reference data and the results of the tests and literature data the reasons. Recommendations on the basis of which can be increased traffic safety and avoid emergency situations of axle box node of possible defects and predict the residual labor resource. Additional studies to further directions. The final work of 3 basic parts, conclusions, suggestions and references. The scope of the work to 74 p. text without additives, 49pictures, 12 tables, 29 bibliographical entries.

På den svenska marknaden står bag-in-box-vin för över 50 procent av allt vin som säljs och Sverige är det land där bag-in-boxen fått starkast fäste i världen (Fremin, 2012). Det gör att bag-in-box-förpackningen är en stor och betydelsefull produkt för vintillverkare och att ha kunskap om säljande design är viktigt för grafiska formgivare samt övriga involverade företag. Den föreliggande studiens syfte var att undersöka vilka designfaktorer på bagin- box-förpackningar som påverkar konsumenternas val av vin i Sverige. Studien innefattade en visuell kartläggning som ämnade ge en överblick av utbudet på ett Systembolag. Detta genomfördes för att skapa en grund till den webbaserade enkätundersökningen som genomfördes under våren 2015. Resultaten visade att majoriteten av respondenterna valde vin baserat på förpackningens utseende och ansåg att de påverkades främst av färg, typsnitt, illustrationer och varumärken. Resultaten visade också att ju högre vinkunskap konsumenterna hade desto mindre påverkade designen deras val. Kvinnor och män hade generellt olika åsikter angående hur stor påverkan de olika designfaktorerna hade vid valet av bag-in-box-vin. Denna studie är endast ett avstamp till ett ämne som innefattar många påverkande faktorer. Det betyder att mer forskning krävs inom ämnet innan helt sanningsenliga resultat kan fastställas. / In the Swedish market the bag-in-box wine stands for over 50 percent of all wine that sells and Sweden is the country where the bag-in-box have the strongest hold in the world (Fremin, 2012). This makes bag-in-box packaging a major and significant product for winemakers. Therefore having knowledge about selling design is important for graphic designers and other involved companies. This study included a visual content analysis of the bag-in-box wine in a liquor store in Sweden in order to get an overview of the supply. With the results, a web-based survey was created and implemented in the spring of 2015. The results showed that the majority of the respondents chose the wine based on the appearance of the package and felt that they were mainly effected by the colour, font, illustrations and brand. Consumers with higher knowledge about wine were less effected by the design in their choice of wine. Women and men had generally different opinions about how much impact the different design factors had in the choice of bag-in-box wine. This study is only a starting point for a subject that includes many influencing factors. Further research is therefore needed before completely truthful results can be established.

Ribonucleoprotein complexes are responsible for some of the post-transcriptional modifications of RNA that occur within the cell, including 2'-O-methylation and pseudouridylation. These modifications contribute, among other things, to RNA folding, inhibition of degradation, and general cellular viability. In this study, we identify residues within the proteins of these complexes that are important to the functioning of the Box C/D and Box H/ACA complexes. Candidates were selected based on previous work and mutant versions of the proteins were introduced in-vivo. Assays were done to determine the functionality of the mutant complex. This work is divided into three parts, focused on the three proteins investigated. The first part is concerned with Nop5, a protein in the Box C/D RNP complex. Nop5 is known to interact with all other proteins and RNAs in the complex, and is believed to serve a primarily structural role, aligning the other components. Mutagenesis study of suspected significant amino acids in this protein showed that it is difficult to disrupt the operation of Nop5 with single changes, but is possible with more extensive mutation. The second part concerns Fibrillarin, the catalytic protein of the Box C/D ribonucleoprotein complex. Previous mutagenesis work identified several important amino acids involved with AdoMet transfer and complex formation. The methylation ability of these mutant complexes were further examined in this work by confirming that the same modification, or lack thereof, occurred at a second rRNA position. The final part of this work is about Nop10, part of the Box H/ACA complex. This work is only preliminary, but begins the process of testing suspected essential amino acids in the structure.