Small-molecule drugs arise as a prospective area to treat different types of cancer. A promising target is the NSD protein family. These proteins have been related to cancers like myeloid leukemia, multiple myeloma, prostate, lung, and breast cancer. However, their treatment is limited to chemotherapy, radiotherapy, and surgical operation that could affect the patient's life quality. In 2020, Huang and collaborators developed a novel kind of inhibitor for NSD1 protein, BT5. This inhibitor covalently binds to the SET domain of the NSD family proteins. However, there is a high affinity for NSD1 than their counterparts. These proteins share a similar structure, but their dynamics could explain the affinity difference. In this project, we compare the NSD family protein dynamics by measuring NMR relaxation experiments. We identify a higher percentage binding for NSD1 and NSD3 to BT5 than NSD2. We also determine the perturbed chemical shifts under the presence of BT5 in NSD1, where the most affected regions are the SET and post-SET domain (auto-inhibitory loop) and the beginning region of the AWS domain. By comparing different NMR relaxation measurements, we identify that the three proteins share high dynamics in the auto-inhibitory loop region, especially in NSD1, and in the AWS domain for NSD1 and NSD3. These motions corresponds to the obtained results by adding BT5 in NSD1, which could indicated a relationship between the AWS dynamics and the auto-inhibitory loop, and the protein affinity.

Made available in DSpace on 2015-03-05T18:40:07Z (GMT). No. of bitstreams: 0 Previous issue date: 29 / Nenhuma / O tópico de desenvolvimento de novos serviços (NSD – New Service Development) tem recebido bastante atenção nos últimos anos por sua importância na geração de diferencial competitivo para as empresas de serviços. No entanto, sua investigação empírica nas organizações tem sido lenta em função da escassez de escalas e itens de medição confiáveis e válidos. Recentemente, um constructo multidimensional denominado de competência em NSD foi proposto para refletir a capacidade da organização de utilizar recursos e rotinas, usualmente em combinação, a fim de atingir um determinado resultado no lançamento de um novo serviço. O constructo foi operacionalizado através de cinco escalas complementares entre si (foco no processo de NSD, acuidade de mercado, estratégia de NSD, cultura de NSD e experiência em TI) e validada em uma amostra de bancos de varejo americanos. A fim de analisar a validade externa da nova escala, esse estudo utilizou uma amostra oriunda de uma empresa brasileira prestadora de serviços de telecomuni / The topic of NSD (New Service Development) has received a lot of attention in the past years, especially given its important competitive necessity in many service industries. However, NSD empirical investigations have been hindered by the lack of sound measurement items and scales. Recently, a second-order multidimensional construct called NSD competence has been proposed to reflect an organization´s expertise in deploying resources and routines, usually in combination, to achieve a desired new service outcome. The construct was operationalized through five complementary multi-item scales (NSD process focus, market acuity, NSD strategy, NSD culture and information technology experience) that were validated in a sample of American retail banks. In order to assess the external validity of the NSD competence construct in a different cultural setting, a sample obtained from key informants of a specific Brazilian telecommunication service provider was used. The results showed a partially satisfactory construct

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Molecular, 2014. / Submitted by Ana Cristina Barbosa da Silva (annabds@hotmail.com) on 2014-10-30T14:14:14Z No. of bitstreams: 1 2014_MarthaSilvaEstrela.pdf: 2513527 bytes, checksum: c69499a20e04f540457a226399f7f1f9 (MD5) / Approved for entry into archive by Tania Milca Carvalho Malheiros(tania@bce.unb.br) on 2014-10-30T15:07:03Z (GMT) No. of bitstreams: 1 2014_MarthaSilvaEstrela.pdf: 2513527 bytes, checksum: c69499a20e04f540457a226399f7f1f9 (MD5) / Made available in DSpace on 2014-10-30T15:07:03Z (GMT). No. of bitstreams: 1 2014_MarthaSilvaEstrela.pdf: 2513527 bytes, checksum: c69499a20e04f540457a226399f7f1f9 (MD5) / O câncer colorretal (CCR) é o terceiro tipo de câncer mais comum entre os homens e o segundo entre as mulheres em todo o mundo. O rastreamento regular pode prevenir muitos casos de CCR e resultar na detecção e diagnóstico precoce, quando pode ser altamente curável. Vários fatores de risco tem sido identificados na gênese do CCR, porém nenhum de forma isolada explica completamente a carcinogênese do cólon e reto. O CCR se desenvolve ao longo de décadas e consiste em uma série de alterações genéticas e epigenéticas que conduzem à transformação maligna. Algumas das alterações genéticas já descritas relacionadas com a carcinogênese colorretal envolvem os genes KRAS, p53 e elementos da via de sinalização TGF-β, como TGFBR2 e SMAD4. Alterações epigenéticas tem se mostrado cada vez mais relevantes na gênese de vários tumores, e tem como característica básica não causar modificações na sequência do DNA, mas sim promover expressão seletiva dos genes dependente do nível do empacotamento do DNA. A metilação de histonas é um dos principais e mais estudados eventos epigenéticos. SETD e NSD são famílias de metiltransferases composta por 10 e 3 genes, respectivamente, que codificam proteínas com domínio SET. Este domínio está envolvido na metilação de resíduos de lisina em histonas e proteínas não-histonas. Até o momento, pouco se sabe sobre a relação de genes das famílias SETD e NSD com a carcinogênese colorretal. No presente estudo foi realizada a avaliação de expressão relativa de todos os genes das famílias SETD e NSD em amostras de tecidos tumorais e normais de pacientes portadores de CCR através de técnica de PCR em tempo real. O perfil de expressão comparativa dessas famílias de genes foi correlacionado com as informações clínico-patológicas de maior relevância prognóstica, disponíveis dos pacientes. Notamos que os genes SETD1A e SETD6 encontram-se significativamente superexpressos nas amostras tumorais de pacientes com até 60 anos quando comparadas às amostras tumorais de pacientes acima de 60 anos. Esses dois genes também apresentaram expressão correlacionada nesse grupo avaliado. Quando analisamos apenas as amostras tumorais de pacientes com invasão perineural não evidenciada, comparadas às amostras tumorais de pacientes com invasão perineural evidenciada, observamos superexpressão do gene NSD3. Em análise pareada de amostras de tecidos normal e tumoral verificamos a hipoexpressão dos genes SETD2 e SETD8 nas amostras tumorais. Esses dois genes também possuem expressão correlacionada nesses tecidos. Desta forma, nossos dados sugerem NSD3 como um possível marcador molecular de melhor prognóstico para pacientes diagnosticados com CCR, além de SETD2 e SETD8 como um possível alvo molecular para o diagnóstico do CCR. ______________________________________________________________________________ ABSTRACT / Colorectal cancer (CRC) is the third most common type of cancer among men and the second among women worldwide. Regular screening can prevent many cases of CRC and result in early detection and diagnosis, as can be highly curable. Several risk factors have been identified in the genesis of CRC, but none alone explains the carcinogenesis in colon and rectum. CRC develops over decades and consists of a series of genetic and epigenetic alterations that lead to malignant transformation. Some of these genetic changes related to colorectal carcinogenesis involves KRAS, p53 and signaling pathway components of the TGF-β as TGFBR2 and SMAD4. Epigenetic changes have been shown to be increasingly important in the genesis of many tumor types, and its basic characteristic is not cause changes in the DNA sequence but also induce selective gene expression dependent on DNA packaging level. Histone methylation is one of the leading and most studied epigenetic events. SETD and NSD methyltransferase families are composed of 10 and 3 genes, respectively, encoding proteins with SET domain. This domain is involved in methylation of lysine residues on histones and non-histone proteins. So far, little is known about the relation of SETD and NSD genes with colorectal carcinogenesis. In this study we evaluated relative expression of all genes of SETD and NSD families in samples of normal tissue and tumor tissue of patients with CRC through real time PCR. The comparative expression profile of these gene families was correlated with clinical and pathological information of greater prognostic relevance available from patients. Here we show that SETD1A and SETD6 genes are significantly overexpressed in tumor samples from patients younger than 60 years when compared to tumor samples from patients over 60 years. These two genes also showed correlated expression in this evaluated group. When we analyzed only tumor samples from patients without perineural invasion, compared to tumor samples from patients with perineural invasion, we observed overexpression of NSD3 gene. In paired analysis of normal tissue samples and tumor tissues samples we verified low expression of SETD2 and SETD8 genes in tumor samples. These two genes also have correlated expression in these tissues. Thus, our data suggested NSD3 as a possible molecular marker of better prognostic for patients diagnosed with CRC, and SETD2 and SETD8 as a possible molecular target for the diagnostic of CRC.

This thesis deals with the modifications of the framework designed for DNS servers testing. Framework is developed by NIC.CZ association and is used primarily for testing the DNS server Knot DNS. The aim of this work are modifications of the framework that will allow simpler testing with this framework, such as: support for multiple implementations of DNS servers, parallel testing, components dummy server and box-in-the-middle, division into multiple components and overall modification of the existing framework. Introduction of thesis is dedicated to the authoritative DNS servers and to the foundations of testing. The remaining part of the thesis deals with the state of the existing framework and the state and testing of modified framework.

Tato práce se zabývá analýzou současných open source řešení pro zabezpečení DNS zón pomocí technologie DNSSEC. Na základě provedené rešerše je navržena a implementována nová knihovna pro použití na autoritativních DNS serverech. Cílem knihovny je zachovat výhody stávajících řešení a vyřešit jejich nedostatky. Součástí návrhu je i sada nástrojů pro správu politiky a klíčů. Funkčnost vytvořené knihovny je ukázána na jejím použití v serveru Knot DNS.

La régulation et le contrôle-qualité de l'expression génique permettent respectivement de maintenir un équilibre entre synthèse et dégradation des ARNm répondant aux besoins cellulaires et d'empêcher l'expression d'ARNm ou protéines aberrants potentiellement toxiques. Pour mieux comprendre ces processus cytoplasmiques, je me suis intéressée à Jlp2, Tac4 et Ska1, trois protéines ayant des liens physiques ou fonctionnels avec des acteurs du contrôle-qualité des ARNm et peptides appartenant aux complexes RQC et SKI. Jlp2 montre des liens de létalité synthétique avec les complexes RQC et SKI mais son absence n'altère pas le " NonStop mRNA Decay ". Elle pourrait donc être impliquée dans une autre voie de contrôle dépendante des complexes RQC et SKI. Tac4 est une ARN hélicase putative associée aux ribosomes, au niveau de l'hélice H16 de l'ARNr 18S comme son homologue putatif mammifère DHX29. Elle interagit également au niveau de régions 3'UTR d'ARNm. Ces observations suggèrent que Tac4 pourrait être impliquée dans la réinitiation de la traduction et le sauvetage de ribosomes non-dissociés récemment identifiés dans la région 3'UTR d'ARNm. Enfin, nous avons identifié Ska1, une protéine appartenant à une nouvelle sous-population de complexes SKI. Nos données suggèrent que ce complexe SKI-Ska1 est impliqué dans la dégradation de transcrits dépourvus de ribosome. Nous proposons un modèle selon lequel ce complexe SKI-Ska1 agirait durant la dégradation de 3'UTR avec l'exosome, puis en arrivant dans la région codante et en rencontrant un ribosome, Ska1 se dissocierait du complexe pour lui permettre d'interagir directement avec le ribosome et poursuivre la dégradation 3'-5' de l'ARN. / Mechanisms responsible for the regulation of gene expression and its quality-control are required, respectively for maintaining an equilibrium between mRNA synthesis and degradation and to prevent synthesis of aberrant mRNAs and proteins potentially toxic for the cells. To better understand these quality-control processes, I studied three factors, Jlp2, Tac4 and Ska1, with physical or functional links described with factors involved in mRNA and protein quality-control, the RQC and SKI complexes. Jlp2 shows synthetic lethality with the RQC and SKI complexes but its deletion has no effect on the NonStop mRNA Decay, suggesting that Jlp2 could be implicated in another control pathway linked to the RQC and SKI complexes. Tac4 is a putative RNA helicase bound to ribosomes, on the 18S rRNA H16 helix, as its mammalian putative homolog DHX29. DHX29 plays a role in translation initiation but surprisingly, Tac4 interacts, in addition to ribosomes, with mRNA 3’UTRs. These observations suggest that Tac4 could be implicated in translation reinitiation and rescue of non-dissociated-ribosomes, recently described within mRNA 3’UTRs. Finally, we identified Ska1, a new factor associated to a SKI complex subpopulation. Our observations suggest that the SKI-Ska1 complex is implicated in the degradation of transcripts devoid of ribosomes. It suggests a model by which the SKI complex would proceed in two steps. First, the SKI-Ska1 complex could assist the exosome to degrade 3’UTR regions of RNAs and then, when its reaches the coding region and encounter a ribosome, Ska1 would leave the complex and allow it to interact directly with ribosomes to proceed further in the 3’-5’ RNA degradation.

The master’s thesis deals with Domain Name System (DNS) and its practical use. It describes hierarchy of domain names, resource record types, protocol used, as well as DNSSEC extension. The most utilized implementations of authoritative and recursive DNS servers are presented. Virtualization, containers and other tools with potential use in labs are described. Two lab scenarios in virtualized environment with instructions for students were designed and developed in this thesis.

Ces travaux sont du domaine de génie systèmes industriels et l'ingénierie de l'innovation. Ils se sont déroulés dans l'industrie de services financiers au sein du groupe de banque-assurance Groupama. Les services représentent 64% de PIB mondial. Le secteur employait en 2007 en France près de 20 millions de personnes contre 5 millions pour l'industrie. Pourtant la recherche sur les processus d'innovation présente un déséquilibre avec un article scientifique NSD (new service development) pour quatre NPD (new product development). L'objectif de ces travaux est de proposer une méthode de pilotage des projets d'innovation dans les services. Pour formaliser le pilotage d'innovation dans les services tout en préservant la zone de liberté nécessaire à l'innovation, nous sommes basés sur un formalisme de type NPD pour définir un processus NSD qui respecte la flexibilité spécifique de l'innovation dans les services. Le coeur de notre hypothèse a été d'identifier des invariants de processus NSD. Nous proposons six classes d'invariants : les OICs (Objets Intermédiaires de Conception), les ressources, les compétences, les tâches, les indicateurs et les méthodes. Leurs interactions sont rendues dans un Diagramme de Classes UML. Un projet peut être décrit comme une « somme » d'OIC eux-mêmes résultat de l'agencement des 5 autres invariants. Ces six classes d'invariants ont été validées sur projets de Groupama. Elles permettent de décrire, suivre, capitaliser, réutiliser des savoirs acquis sur des projets antérieurs et de manager les projets innovants. Nous proposons aussi un processus de pilotage des projets NSD, formalisé par un Diagramme d'Ordonnancement des Phases sous MEGA / The present thesis is about the field of system engineering and innovation engineering. It took place in the financial industry within the group of banking insurance Groupama. Services represent 64% of world's GDP (Gross Domestic Product). This branch employed in 2007 in France about 20 million people against 5 million for industry. Still the research on the innovation processes presents a gap with only one NSD (new services development) article against four NPD (new products development). The aim of this work is to propose a method for management of innovation projects in the services branch. To formalize the management of innovation projects for the services, while preserving the space required for innovation, we use a NPD's type formalism in order to define a NSD process while respecting the specific flexibility of the innovation in the services branch. The center of our hypothesis was to identify invariants within the NSD process. We propose six classes of invariants: IDOs (Intermediate Design Objects), the resources, the skills, the tasks, the indicators and the methods. Theirs interactions are shown in a UML Diagram of Classes. A project may be represented as an "addition" of IDOs, where they are the result of 5 other invariants arrangements. These six classes of invariants were validated on Groupama projects. They are relevant to describe, monitoring, capitalize, re-use of the knowledge acquired on previous projects and to manage innovative projects as well. We also propose a process of piloting of the NSD projects, formalized by a Diagram of Phases Sequencing of MEGA

Submitted by Silvana Teresinha Dornelles Studzinski (sstudzinski) on 2015-05-08T15:58:07Z No. of bitstreams: 1 Debora Moehlecke.pdf: 980082 bytes, checksum: fdb15528a7aada1f0dafab6b637180c8 (MD5) / Made available in DSpace on 2015-05-08T15:58:07Z (GMT). No. of bitstreams: 1 Debora Moehlecke.pdf: 980082 bytes, checksum: fdb15528a7aada1f0dafab6b637180c8 (MD5) Previous issue date: 2014-06-09 / Nenhuma / Para que as empresas possam sobreviver e principalmente prosperar, novos serviços são desenvolvidos, por vezes aproveitando a ideia de um cliente ou mesmo uma oportunidade percebida no mercado. O desenvolvimento de novos serviços (DNS) já foi visto como um mero acontecimento que se tornava possível devido à sorte ou intuição. Porém, com o aumento dos estudos sobre o tema, o conhecimento foi ampliado e atualmente defende-se a necessidade de planejamento do DNS e uma maior compreensão gerencial para que assim as empresas possam alcançar o desempenho esperado. A fim de buscar entendimento da capacidade das empresas em utilizar recursos e rotinas para obter sucesso no novo serviço lançado no mercado, em um estudo anterior foi elaborado um modelo de competência em DNS considerando cinco dimensões: i) Foco no processo de DNS; ii)Acuidade de mercado; iii)Estratégia de DNS; iv)Cultura de DNS; e v)Experiência em Tecnologia da Informação (TI). Utilizando este modelo, o presente estudo visa verificar a relação entre as dimensões formadoras da competência em desenvolvimento de novos serviços e o desempenho das empresas no desenvolvimento destes novos serviços, por meio de hipóteses de pesquisa. Tendo em vista a importância do setor de serviços intensivos em conhecimento para a economia mundial e o crescimento destas empresas no Brasil, buscou-se através de uma survey com 239 organizações, confirmar a relação que há entre as cinco dimensões no DNS, validar o instrumento de “modelo de competência” para o desenvolvimento de novos serviços e também verificar como as dimensões contribuem em conjunto para o desempenho das empresas de serviços intensivos em conhecimento (ESIC). Por meio de testes estatísticos como a técnica de análise fatorial exploratória e a análise de regressão múltipla, verificou-se que quatro fatores podem ser considerados responsáveis por 30% do desempenho em DNS das empresas analisadas. Destaca-se que a dimensão Acuidade do mercado foi considerada a de maior efeito em desempenho, seguida da Experiência em TI, Cultura de DNS e Foco no processo de DNS, entretanto a dimensão Estratégia de DNS não pode ser avaliada pois suas variáveis foram eliminadas durante a análise fatorial exploratória. / In order to enable companies to survive and succeed in the market, new services are developed, sometimes availing some customer idea oreven a perceived market opportunity. New service development (NSD) had been seen as a result of luck or intuition. However, due to the increase of studies on the subject, the knowledge has been expanded and now is believed the need of NSD planning and greater managerial understanding so that companies can achieve the expected performance. In order to seek understanding of the ability of firms to use resources and routines to succeed in the new service launched in the market, on a previous study a NSD competence model was developed considering five dimensions: i)NSD process focus; ii)Market acuity; iii)NSD strategy; iv)NSD culture and; v)Information Technology (IT) experience. Using this model, the present study aims to investigate the relationship between all competence dimensions in new service development and the performance of some companies in developing new service through research hypotheses. Given the importance of knowledge-intensive service sector in the world economy and the growth of these companies in Brazil, we attempted through a survey with 239 o rganizations to confirm the relationship between these five dimensions during the new service development, also to validate the instrument “model of NSD competence” as well as to check how all dimensions contribute to the knowledge-intensive service company performance. By statistical tests such as the technique of exploratory factor analysis and multiple regression analysis, it was found that four dimensions can be considered responsible for 30% NSD performance. It is noteworthy that market acuitywas found to have more effect on performance, followed by IT experience, NSD culture and NSD process focus. However, NSD strategycould not be evaluated as their variables were eliminated during exploratory factoranalysis.

Ces travaux sont du domaine de génie systèmes industriels et l'ingénierie de l'innovation. Ils se sont déroulés dans l'industrie de services financiers au sein du groupe de banque-assurance Groupama. Les services représentent 64% de PIB mondial. Le secteur employait en 2007 en France près de 20 millions de personnes contre 5 millions pour l'industrie. Pourtant la recherche sur les processus d'innovation présente un déséquilibre avec un article scientifique NSD (new service development) pour quatre NPD (new product development). L'objectif de ces travaux est de proposer une méthode de pilotage des projets d'innovation dans les services. Pour formaliser le pilotage d'innovation dans les services tout en préservant la zone de liberté nécessaire à l'innovation, nous sommes basés sur un formalisme de type NPD pour définir un processus NSD qui respecte la flexibilité spécifique de l'innovation dans les services. Le cœur de notre hypothèse a été d'identifier des invariants de processus NSD. Nous proposons six classes d'invariants : les OICs (Objets Intermédiaires de Conception), les ressources, les compétences, les tâches, les indicateurs et les méthodes. Leurs interactions sont rendues dans un Diagramme de Classes UML. Un projet peut être décrit comme une " somme " d'OIC eux-mêmes résultat de l'agencement des 5 autres invariants. Ces six classes d'invariants ont été validées sur projets de Groupama. Elles permettent de décrire, suivre, capitaliser, réutiliser des savoirs acquis sur des projets antérieurs et de manager les projets innovants. Nous proposons aussi un processus de pilotage des projets NSD, formalisé par un Diagramme d'Ordonnancement des Phases sous MEGA.