Am 24. Oktober 2011 wurde in ganz Deutschland der „Tag der Bibliotheken“ begangen. Im Anschluss daran startete zum vierten Mal die bundesweite einwöchige Aktionswoche unter dem Motto „Treffpunkt Bibliothek – Information hat viele Gesichter“. In Sachsen waren zahlreiche Bibliotheken mit einem bunten Programm dabei.

Retrotransposon mobilization is a major source of genome evolution. However, the functional consequences of these events, and particularly their influence upon transcriptional activity, are poorly defined. The extent of retrotransposon transcription, as well as that of other repetitive elements, has eluded systematic study due to difficulties in discriminating elements copied in multiple genomic loci. Moreover, the potential regulatory effects of retrotransposon transcription upon the expression of neighbouring protein-coding genes are also largely unknown. This thesis develops methods to survey repetitive element expression and assess the functions of retrotransposons in the mouse and human genomes. Chapter 1 summarises the complex transcriptional output of the mammalian genome, the functional annotation of this expression and the genomic and bioinformatic tools available for its detection. Chapter 2 explores the capacity of short sequence tags to discern transcription from individual repetitive elements, as well as from protein-coding genes. It is based upon a publication that critiqued the bioinformatics associated with Cap Analysis Gene Expression (CAGE) and developed novel methodologies to resolve repetitive element transcription. Chapter 3 describes the development of an updated CAGE mapping pipeline for the fourth stage of the international Functional Annotation of Mouse (FANTOM) project, which lead to the generation of a research article and a book chapter. These works demonstrated the enhanced utility of CAGE when coupled with next-generation sequencing, highlighted the benefits of CAGE when applied to systems biology and profiled the temporal expression of human repetitive elements. Chapter 4 presents an in-depth analysis of repetitive element transcription in the mouse and human genomes. Using CAGE, approximately 250,000 retrotransposon associated transcription start sites were defined, many of which were tissue-specific. Retrotransposons were found to frequently function as alternative promoters for protein-coding genes and/or express non-coding RNAs. Furthermore, when retrotransposons were found within the 3’UTR of protein-coding genes, there was strong evidence for the reduced expression of the corresponding transcripts. A genome-wide screen for strong expression correlation between repetitive elements and neighbouring protein-coding genes identified approximately 23,000 candidate regulatory regions derived from retrotransposons, including several hundred putative boundary elements. These were in addition to more than two thousand examples of bidirectional transcription found in retrotransposons, which are known to be a source of double stranded RNAs involved in RNA interference. Chapter 5 explores the proportion of the mouse embryonic stem cell transcriptome comprised of repeat-derived transcripts, using next-generation RNA sequencing. This study defined the dynamic expression of repetitive elements at the greatest resolution achieved to date and demonstrated that repetitive elements are an intrinsic part of the mammalian transcriptional landscape.

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No. of bitstreams: 1 Tese - Marcicleide Lima do Espirito Santo.pdf: 7339955 bytes, checksum: 22519cc9c5e5f14e2991f1e20ddb938f (MD5) Previous issue date: 2006-06-22 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / The camu-camu (Myrciaria dubia (H.B.K.) McVaugh) is a native sort of the Amazonian region, whose fruit presents elevated content of ascorbic acid (vitamin C). The study of the functional genome in camu-camu fruits has like base the expressed sequence tags sequencing - ESTs. Faced with the displayed the present thesis is going to analyze and identify express genes in camu-camu fruits by means of ESTs sequencing. The total RNA was extracted from the shell-pulp. The extremity 5’sequencing' of cDNA insert was carried out so much in the Technology of the DNA Laboratory (UFAM) and in the Sequencing Platform (EMBRAPA/CENARGEN). The ESTs sequences obtained were submitted to the System Genome, program of genomic annotation that integrates analysis management programs and viewing of nucleotides sequences. It developed an efficient procedure for total RNA extraction of camu-camu fruits that enabled the obtaining of mRNAs of quality, utilized in the making of cDNAs of sizes varied (500pb to 4Kb). From the sequencing were obtained 3196 ESTs valid, being formed 1546 singletons and 358 contigs, resulting of 2586 ESTs sequences in total with similarity the sequences found in the gene bank. The analysis library clusterization revealed an index of 81% novelty and 32,54% redundancy. Around 90% of the contigs presented decrease redundancy (2-4 reads by contigs). The facts of the categorization of the proteins identified detached the posttranslational modification, protein turnover, chaperones (13,2%) category. From the hoist of the species with bigger number of ESTs with similarity the camu-camu sequences detached itself Arabidopsis thaliana with 49%. Around 10 uniques presented very high similarity (and-value 0.0) to known genes. The ESTs more abundantly express in camu-camu fruits encode to gluthatione s-transferase. They were observed around 3% sequences (97 ESTs) with decrease similarity (e-value > e-10) and 15% did not they present similarity with no contained sequence in the gene bank. They were identified 138 ESTs sequences (4,3%) that they encode molecular chaperones with prevalence of the sHSP family that represents 33% of express chaperones. ESTs related to the ascorbic acid metabolism also were identified, being nine related the synthesis and six come back for ascorbic acid conversion and recycling. ESTs related to the ripening and mechanisms of defense of the fruit also were noticeable. / O camu-camu (Myrciaria dúbia (H.B.K.) McVaugh) é uma espécie nativa da região Amazônica, cujo fruto apresenta elevado teor de ácido ascórbico (vitamina C). O estudo do genoma funcional em frutos de camu-camu tem como base o seqüenciamento de fragmentos de seqüências expressas - ESTs (Expressed Sequence Tags). Diante do exposto a presente tese pretende analisar e identificar genes expressos em frutos de camu-camu por meio de seqüenciamento de ESTs. O RNA total foi extraído a partir da casca-polpa. O seqüenciamento da extremidade 5’ de insertos de cDNA foi realizado tanto no Laboratório de Tecnologia do DNA da UFAM e como na Plataforma de Seqüenciamento da EMBRAPA/CENARGEN. As seqüências ESTs obtidas foram submetidas ao Sistema Genoma, programa de anotação genômica que integra programas de gerenciamento de análise e visualização de seqüências nucleotídicas. Os resultados obtidos foram o desenvolvimento de um procedimento eficiente para extração de RNA total de frutos de camu-camu que possibilitou a obtenção de mRNAs de qualidade, utilizados na confecção de cDNAs de tamanhos variados (500pb a 4Kb). A partir do seqüenciamento foram obtidas 3196 ESTs válidas, sendo formados 1546 singletons e 358 contigs, resultando num total de 2586 seqüências ESTs com similaridade a seqüências encontradas no banco de genes. A análise da clusterização da biblioteca revelou um índice de 81% de novidade e 33% de redundância. Cerca de 90% dos contigs apresentaram baixa redundância (2-4 reads por contigs). Os dados da categorização das proteínas identificadas destacaram a categoria modificação pós-traducional, proteína turnover, chaperonas (13,2%). A partir do levantamento das espécies com maior número de ESTs com similaridade a seqüências de camu-camu destacou-se Arabidopsis thaliana com 49%. Cerca de 10 uniques apresentaram altíssima similaridade (e-value 0.0) a genes conhecidos. Os ESTs mais abundantemente expresso em frutos de camu-camu codificam a glutationa s-transferase. Foram observados cerca de 3% de seqüências (97 ESTs) com baixa similaridade (e-value > e-1010) e 15% não apresentaram similaridade com nenhuma seqüência contida no banco de genes. Foram identificadas 138 seqüências ESTs (4,3%) que codificam chaperonas moleculares com destaque à família sHSP que representa 33% das chaperonas expressas. ESTs relacionados ao metabolismo do ácido ascórbico também foram identificados, sendo nove relacionados a síntese e seis voltados para conversão e reciclagem do ácido ascórbico. ESTs relacionados ao amadurecimento e mecanismos de defesa do fruto também foram destacados.

Made available in DSpace on 2014-06-12T22:56:34Z (GMT). No. of bitstreams: 2 arquivo1229_1.pdf: 2037381 bytes, checksum: 76ef1d886b39ab00e1420e5b4c061499 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2011 / Propusemos nesse estudo que a Psicologia pode contribuir para a produção de conhecimento na área de Design e, para isso, nos apoiamos na perspectiva pragmática para analisar a produção de sentidos sobre tag clouds. Este termo refere-se a um recurso criado para representar o processo de tagging, de atribuir palavras-chave aos conteúdos da web, frequentemente utilizado em blogs. Objetivamos, então, identificar os sentidos produzidos por blogueiros sobre tag clouds a respeito dos motivos de incorporação desse recurso nos blogs e dos padrões de utilização. Para a construção de dados, pesquisamos por posts que discutiram sobre a utilização de tag clouds, visando a identificação dos sentidos sobre os motivos de incorporação; e, por outro lado, para investigar os sentidos sobre os padrões de utilização, convidamos blogueiros para a participação de entrevistas sobre o tema. Encontramos que os sentidos produzidos sobre os motivos de incorporação são direcionados a comparações intrarecurso, entre tipos de tag clouds, e inter-recurso, entre tag clouds e outros recurso como lista de tags e menu. Em relação aos padrões de utilização identificamos que estes estiveram relacionados aos sistemas de informação hospedeiros dos blogs. No 'Blogger.com', tag clouds caracterizam-se como menus para navegação nas páginas pessoais e, em geral, os blogueiros optaram por utilizá-lo nos blogs, e, por outro lado, no 'Wordpress.com', os blogueiros, em sua maioria, não optaram por utilizar tag clouds em suas páginas, já que as tags criadas direcionam para a navegação no sistema de informação. Concluimos, então, que representar tags através de tag clouds no próprio blog significa utilizar esses dados parcialmente, isto é, considerando apenas seu aspecto individual. Contudo, ao representar o conjunto de tags de um sistema de informação em uma única tag clouds, prioriza-se o aspecto coletivo. Porém, as duas dimensões, individual e coletiva, não são excludentes e ao precisar optar por uma dessas, os blogueiros são afastados do que parece ser a característica e função diferenciadora de tags e tag clouds na atividade, isto é, a união entre informações individuais e coletivas, entre interesses e conhecimentos que falam sobre um sujeito e, ao mesmo tempo, sobre uma comunidade

This thesis deals with RFID tags design. It comprises an overview of the RDIF technology while focusing upon the passive tags in UHF band. The CST Microwave studio software has been used for draft simulations. The resulting tags have been subjected to parameter measuring – the reflection coefficient, the minimum capacity needed to switch on the tag, the maximum reading distance. The influence of the proximity of metal objects and other materials has also been examined.

In the thesis LIFE ACCORDING TO THEM I deal with beauty blogs on YouTube. I use my own pieces of work, especially video recordings of performances for the camera or for the audience, which were created during the entire master’s degree study. Their further processing together with ideas and experiences leads to an audio-visual collage, that will accompany the final performative lecture. The aforementioned collage may also exist as a separate video-essay expressing my experiences with the topic of blogging.

Graffiti tags are similar to the common human signatures. This report analyses, which signature recognition methods could be useful in the field of preventing graffiti vandalism. It also discusses differences between signatures and graffiti tags.

MicroRNAs are small non-coding RNAs that regulate gene expression through RNA interference. These small RNAs enact gene silencing by forming a RNA-inducing silencing complex together with the effector protein Argonaute. The function of the Argonautes in the social amoeba Dictyostelium discoideum is not yet fully understood. In this study, we look closer at Argonaute B by investigating if it is possible to extract the protein from the cells by the addition of a polypeptide protein tag called 3xFlag. At the same time, we also look into if Argonaute B is important for cell growth. Sequences of the 3xFlag tag with or without the Argonaute B gene (agnB) attached had previously been cloned into a vector and transformed into Dictyostelium discoideum cell. The 3xFlag::agnB sequence was confirmed in wild type and agnB knock-out strains through polymerase chain reaction. We then verified the expression of the fusion protein in the cells by western blot. The cell growth was measured by how the number of cells changed over time. The experiment suggested that Argonaute B is important for growth. Our result show that the construct 3xFlag::agnB sequenced had correctly been transformed into the strains and is highly expressed under tested conditions. We could also see that Argonaute B is an important factor in cell growth.

Globally, breast cancer is the leading cause of death in the female population aged 45 and below with a breast cancer incidence reaching 18.1 million in the year 2018. Triple negative breast cancer (TNBC) is part of a group of cancers that lack the expression of Progesterone receptor (PR), Estrogen receptor (ER) and Human epidermal growth factor receptor 2 (HER2). TNBC is commonly associated with early stage metastasis with low survival rates as well as a high frequency of recurrence and proves to be problematic in both the young and elderly female populations. Conventional diagnostic methods for TNBCs include mammography, magnetic resonance imaging (MRI) and ultrasound while therapeutic methods include mastectomy and breast conserving surgery (coupled with radiation therapy). The lack of effective therapeutic options, poor prognostic value and high rates of metastasis, has made treatment of TNBC difficult. The major focus of this work was on the following tumour associated antigens (TAAs): CSPG4 (a transmembrane protein found in 50% of TNBC cases), EGFR (which is overexpressed in 13-76% of TNBCs), and MSLN (which is overexpressed in 67% of TNBCs) as potential targets for monospecific therapy. The evolution of antibody-based immunotherapy strategies has led to applications of single chain variable fragment (scFv) & single domain/nanobody (VHH) antibody formats for diagnostic and therapeutic purposes. In this work, these recombinant antibody fragments have been combined with SNAP-tag, a modified version of the human DNA repair enzyme O6-alkylguanine-DNA-alkyltransferase (AGT), which autocatalytically binds benzyl-guanine modified substrates such as fluorophores or small molecule toxins covalently in a 1:1 stoichiometry. In this study, the primary aim was the comparison of different antibody formats fused to SNAPtag and the potential of these biopharmaceuticals towards immunodiagnosis and therapy of TNBCs. First functionalities of two scFv SNAP fusion proteins and one VHH SNAP fusion protein previously not having been described are provided through binding analyses on receptor positive tumour cell lines. This was achieved by in-silico design and molecular cloning of genetically fused antiCSPG4(scFv), -MSLN(scFv), -MSLN(VHH), -EGFR(scFv) & -EGFR(VHH) to SNAP-tag. The final constructs were confirmed by Sanger sequencing and subsequently transfected into a mammalian vector system (HEK293T) for transient expression of the engineered fusion proteins. Full length protein purified from cell culture supernatant was analysed for diagnostic/therapeutic activities dependant on the substrate attached in the form of a fluorophore or small molecule toxin resulting in recombinant antibody-drug conjugates (ADCs). The study shows promise in providing new immunodiagnostic and therapeutic agents that are specific and less harmful than the current state of the art procedure

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