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Ectopic expression and knocking-down of LINE-1 mRNA in human mesenchymal stem cells: impact on in vitro osteogenic and adipogenic differentiationAtinbayeva, Nazerke 05 1900 (has links)
There are two classes of transposable elements: DNA transposons and retrotransposons. DNA transposons spread in the genome by “cut and paste” mechanism. In contrast, retrotransposons use copy and paste strategy involving RNA and retrotranscriptase mediated mechanism; these include long interspersed nuclear elements-1 (LINE-1, L1) and short interspersed nuclear elements (SINE). In mammals, in order to maintain genome integrity both types of transposons are tightly repressed. However, some copies of retrotransposons are still active in germ cells contributing to natural variation. Surprisingly, recent reports indicate that also somatic cells support L1 reactivation in early development, in particular in the brain leading to mosaicism. However, whether L1 retrotransposition is a part of other cell lineage developmental programs and its functional significance in the context of cell differentiation remain to be elucidated.
To address this question, I investigated whether L1 retrotransposition was occurring during in vitro osteogenic and adipogenic differentiation of bone marrow derived human mesenchymal stem cells (hMSCs).
Interestingly, clinical observations have revealed loss of bone density in HIV-infected individuals treated with nucleoside analogs that inhibit HIV retrotranscriptase, as well as the endogenous one encoded by L1s. This observation made us to hypothesize that transposable elements played a positive role in post-natal bone homeostasis.
I found that while adipogenesis is “retrotransposition free”, osteogenic differentiation is a “retrotransposition-prone” process and its inhibition blocks its genetic program. Indeed, L1 DNA content does not change during adipogenic differentiation and that of retrotranscriptase does not have any effect on the acquisition of a terminally differentiated phenotype. In contrast, soon after MSCs commitment into pre-osteoblasts, L1 retrotransposable elements increase their expression and actively transpose. Inhibition of retrotransposition and knock down of L1 mRNA strongly impairs matrix deposition. Moreover, I forced L1 expression in in vitro adipogenesis, by directly delivering L1 mRNA to the cells. Interestingly, overexpression of L1 elements was detrimental for in vitro adipogenesis. Then, I performed loss of function experiments in osteogenesis by directly targeting and degrading the L1 endogenous transcript. This experiment confirmed the positive role of L1 reactivation in the osteogenic context, suggesting also a possible role for L1 RNA, distinct from retrotransposition.
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Proteomic studies on anti-proliferating activities of adenosine and cordycepin in human cancer cell lines.January 2004 (has links)
Tam Wai-Kwan Karen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 109-128). / Abstracts in English and Chinese. / Thesis committee --- p.i / Statement --- p.ii / Abstract --- p.iii / Acknowledgements --- p.vi / Abbreviations --- p.vii / Table of Contents --- p.ix / List of Tables --- p.xii / List of Figures --- p.xiv / Chapter 1. --- Introduction --- p.1 / Chapter 2. --- Literature Review --- p.2 / Chapter 2.1 --- Introduction of Cordyceps --- p.2 / Chapter 2.2 --- Pharmacological functions of Cordyceps --- p.3 / Chapter 2.2.1 --- Functions in respiratory system --- p.3 / Chapter 2.2.2 --- Functions in renal system --- p.7 / Chapter 2.2.3 --- Functions in hepatic system --- p.8 / Chapter 2.2.4 --- Functions in cardiovascular system --- p.9 / Chapter 2.2.5 --- Functions in endocrine and steroid system --- p.10 / Chapter 2.2.6 --- Functions in the immune system --- p.11 / Chapter 2.2.7 --- Functions in nervous system --- p.15 / Chapter 2.2.8 --- Controls in glucose metabolism --- p.15 / Chapter 2.2.9 --- Anti-oxidation activity --- p.16 / Chapter 2.2.10 --- Anti-tumor activity --- p.18 / Chapter 2.3 --- Active ingredients of Cordyceps and their related biological activities --- p.20 / Chapter 2.3.1 --- Polysaccharides --- p.20 / Chapter 2.3.2 --- Nucleosides --- p.21 / Chapter 2.3.2.1 --- Adenosine --- p.21 / Chapter 2.3.2.2 --- Cordycepin --- p.24 / Chapter 2.4 --- Proteomic tools in studies of the change in protein expression --- p.25 / Chapter 2.4.1 --- Two-dimensional electrophoresis --- p.27 / Chapter 2.4.2 --- Mass Spectrometry --- p.28 / Chapter 3. --- Methods and Materials --- p.30 / Chapter 3.1 --- Cell lines and culture conditions --- p.30 / Chapter 3.2 --- Trypan blue exclusion method --- p.30 / Chapter 3.3 --- Cell counting --- p.31 / Chapter 3.4 --- Anti-proliferation assay --- p.31 / Chapter 3.5 --- Anti-proliferation assay of normal cell line --- p.32 / Chapter 3.6 --- Determination of ic50 --- p.33 / Chapter 3.7 --- Sample preparation for proteins studies --- p.33 / Chapter 3.8 --- Protein quantitation --- p.34 / Chapter 3.9 --- Gel electrophoresis --- p.36 / Chapter 3.10 --- Image analysis --- p.37 / Chapter 3.11 --- In-gel digestion and MALDI-ToF MS --- p.37 / Chapter 3.12 --- Statistical Analysis --- p.39 / Chapter 3.13 --- Chemicals --- p.39 / Chapter 4. --- Results --- p.41 / Chapter 4.1 --- MTT assay --- p.41 / Chapter 4.1.1 --- The anti-proliferating activity of adenosine against cancer cell lines (HepG2 and SV7tert) and normal cell line (Hs68) --- p.41 / Chapter 4.1.2 --- The anti-proliferating activity of cordycepin against cancer cell lines (HepG2 and SV7tert) and normal cell line (Hs68) --- p.42 / Chapter 4.1.3 --- The anti-proliferation effects of adenosine and cordycepin --- p.42 / Chapter 4.2 --- Changes in protein expression --- p.50 / Chapter 4.2.1 --- "Corresponding drug treatment of cell lines (HepG2, SV7tert and Hs68)" --- p.50 / Chapter 4.2.2 --- "Comparison of protein profiles from cells (HepG2, SV7tert or Hs68) under the normal and drug treated (with either adenosine or cordycepin) conditions" --- p.51 / Chapter 4.2.2.1 --- HepG2 study --- p.51 / Chapter 4.2.2.2 --- SV7tert study --- p.52 / Chapter 4.2.2.3 --- Hs68 study --- p.52 / Chapter 4.2.3 --- Protein identification --- p.53 / Chapter 4.2.3.1 --- HepG2 cell line --- p.53 / Chapter 4.2.3.2 --- HepG2-changes in protein expressions after adenosine treatment --- p.54 / Chapter 4.2.3.3 --- HepG2-changes in protein expressions after cordycepin treatment --- p.54 / Chapter 4.2.3.4 --- SV7tert cell line --- p.54 / Chapter 4.2.3.5 --- SV7tert-changes in protein expressions after cordycepin treatment --- p.55 / Chapter 4.2.3.6 --- Hs68 cell line --- p.55 / Chapter 4.2.3.7 --- Hs68-changes in protein expressions after cordycepin treatment --- p.56 / Chapter 5. --- Discussion --- p.89 / Chapter 5.1 --- anti-proliferation assays --- p.89 / Chapter 5.2 --- changes in protein expression: --- p.90 / Chapter 5.2.1 --- Protein alterations in HepG2 --- p.91 / Chapter 5.2.1.1 --- Changes in protein expression (membrane protein and transport: Trimethyllysine hydroxylase) --- p.91 / Chapter 5.2.1.2 --- Changes in protein expression (protein synthesis and folding: carboxypeptidase E) --- p.92 / Chapter 5.2.1.3 --- Changes in protein expression (membrane proteins and transport: calumenin and electron transfer flavoproteins) --- p.93 / Chapter 5.2.2 --- Protein alterations in SV7tert --- p.94 / Chapter 5.2.2.1 --- Changes in protein expression (protein synthesis and folding: BiP(GRP78)) --- p.94 / Chapter 5.2.2.2 --- Changes in protein expression (cell defense and tolerance: Hsp60 (chaperonin); TANK binding kinase-1) --- p.96 / Chapter 5.2.2.3 --- Changes in protein expression (metabolism: prolyl 4-hydroxylase; aldolase A; glyceraldehyde-3-phosphate dehydrogenase) --- p.97 / Chapter 5.2.2.4 --- Changes in protein expression (cell growth and division: βII tubulin; HnRNP Al) --- p.100 / Chapter 5.2.3 --- Protein alterations in Hs68 --- p.101 / Chapter 5.2.3.1 --- Changes in protein expression (metabolism: triosephosphate isomerse 1) --- p.101 / Chapter 6. --- Discussion --- p.103 / Chapter 6.1 --- The antiproliferating activities of adenosine and cordycepin --- p.103 / Chapter 6.2 --- "Effects of adenosine and cordycepin on the changes in protein expressions in HepG2, SV7tert and Hs68" --- p.104 / Chapter 6.3 --- Problems and improvements in two-dimensional gel electrophoresis --- p.105 / Chapter 7. --- Conclusion and future prospectives --- p.107 / References --- p.109
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Computer-based Number Categorization as an Intervention for Computer-based Number Line EstimationAwotwi, Ama January 2017 (has links)
The number line is a versatile tool. When used in estimation, it can serve as a visual representation of number. This study evaluated the relationship between sorting numbers by magnitude and number line estimation performance. Fifty-eight participants in Grades 1, 2, and 3 estimated values on a 0-100 number line over four sessions. During two intervention sessions they sorted numbers into 5 categories either linearly or nonlinearly before they estimated the same target values. The linear group’s number line estimates had less error than the nonlinear group’s estimates at posttest. In particular, the participants who started with low numeracy scores in the linear group outperformed their counterparts in the nonlinear group on the number line estimation task. Computer-based number categorization supports computer-based number line estimation skills when numbers are categorized linearly. This finding extends the representational mapping hypothesis to computer-based scaffolds.
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A leitura de jornais impressos e digitais em contextos educacionais : Brasil e Portugal /Arena, Adriana Pastorello Buim. January 2008 (has links)
Orientador: Juvenal Zanqueta Júnior / Banca: Norma Sandra de Almeida / Banca: Raquel Lazzari Leite Barbosa / Banca: Sônia Aparecida Lopes Benites / Banca: Valdir Heitor Barzoto / Resumo: Essa pesquisa parte da existência de um novo fenômeno, a cibercultura, que implementa um novo espaço de leitura, com ferramentas que possibilitam a construção de novas práticas, interferindo na formação do leitor em processo e modificando-a. A leitura do jornal impresso e do jornal on-line é o objeto de análise desta tese. Com o objetivo de compreender e discutir os indícios que se destacam nessas práticas de leitura em contextos educacionais diferentes foram selecionadas seis professoras e trinta alunos, entre 9 e 10 anos, de quatro escolas públicas, sendo uma do Brasil e três de Portugal. Ao estudar duas realidades histórico-culturais diferentes, mas com o uso da mesma língua, a tese tem o objetivo de contribuir para o avanço nas pesquisas em educação e de levantar questões e reflexões sobre novas tecnologias que surgem no contexto social e, particularmente, no escolar. Trata-se de uma investigação de cunho essencialmente qualitativo e interpretativo. Metodologicamente, o trabalho de pesquisa foi dividido em duas partes: a primeira, foi realizada uma experiência de ensino com 6 alunos brasileiros de uma escola pública de Marília, cujo suporte metodológico foi a pesquisa-ação. A metodologia escolhida para o desenvolvimento da primeira parte deste estudo visava a encontrar indícios de práticas de leitura do jornal impresso transportadas para situações de leitura do jornal on-line. As aulas de leitura do suporte eletrônico e do impresso foram gravadas em vídeo e os relatos verbais dos alunos gravados em fitas cassetes. Na segunda parte da pesquisa, um estudo etnográfico, tem como instrumento principal de coleta a observação e a entrevista semi-estruturada, realizada tanto com professores, como com alunos do Brasil e de Portugal, com o propósito de investigar o uso do jornal impresso e on-line... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This research starts from the existence of a new phenomenon, the cyber culture, which implements a new space for reading, with tools that will allow the building of new practices, interfering and modifying the formation of the reader in process. The reading of the print and the on-line newspaper is the object of the analysis of this thesis. With the main goal of understanding and discussing the evidences that stand out in these reading practices, six teachers and thirty students were selected, between 9 and 10 years old, of four public schools, being one from Brazil and three from Portugal, for the development of the research. By studying two different historic-cultural realities, but with the use of the same language, the goal of the thesis is contributing with the advance in educational researches, and raise questions and reflections about new technologies that are brought out on the social and particularly scholar context. It's about an investigation, essentially, qualitative and interpretative. Methodologically, the research job was divided in two parts: in the first part of the research, a study experience was made with 6 Brazilian students of a public school from Marília, whose methodological gadget was the research-action. The chosen method for the development of the second part of this study aimed to find evidences of reading practices of the print newspaper transported to reading situations of the on-line newspaper. The reading classes of the electronic and print supports were recorded in video and the students's verbal reports, taped in video cassette tapes.The second part one is about an ethnographical study, having as main instrument the observation and the semi-structured interview made with teachers as well as students, from Brazil and Portugal, with the purpose of investigating the use of print and on-line newspaper... (Complete abstract click electronic access below) / Doutor
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Analýza klíčových faktorů úspěchu ABradia, a. s. / Achievement´s key factor analysis of ABradio, joint stock companyVoráčková, Lenka January 2006 (has links)
Práce odhaluje stěžejní faktory, jež stojí za úspěšnou aktivitou společnosti ABradio, a. s, která provozuje (kromě jiných činností) veliké množství rádiových stanic po internetu. Zároveň zjišťuje vnitřní i vnější negativní prvky ovlivňující podnik. Stručně se zabývá i konkurencí firmy na čekém trhu. V závěru je navržen postup na odstranění úzkých míst s cílem přispět k lepším výsledkům organizace a způsobu její činnosti uvnitř i navenek.
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Internetová bannerová reklama - nový model on-line nákupu a prodeje bannerové reklamy / Internet banner advertising - a new model of online buying and selling banner advertisementTabáček, Jiří January 2011 (has links)
The first, theoretical part is dedicated to a complete summary of the theory behind the banner advertising in general -- starting at the very beginning of internet advertising in the past, to today's latest forms of internet advertising campaigns. In this part you can also find an exact description of functionality of these models and their possible options. The following, practical part is focused on implementing the new model for buying and selling space for advertising on the web. This part of my Thesis includes analysis of functionality, advantages and disadvantages for the use of the new model and analysis of issues that occurred during its implementation. A questionnaire has been developed and used to analyze this. My Theses concludes with proposed recommendations and future development of the new model.
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Concepção e simulação de célula robotizada para operações de acabamentoBarbosa, João Pedro Marques January 2010 (has links)
Tese de mestrado integrado. Engenharia Mecânica (Opção de Automação). Faculdade de Engenharia. Universidade do Porto. 2010
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Questionários digitais on-line - QDO : sistema de representação, recolha e tratamento de inquéritos em tempo realAntunes, Nuno Gonçalves January 2007 (has links)
Tese de mestrado. Tecnologia Multimédia. Faculdade de Engenharia. Universidade do Porto. 2007
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Ensino rápido de células de quinagem ADIRACoelho, Daniel Alves January 2011 (has links)
Versão final / Tese de mestrado integrado. Engenharia Mecânica. Universidade do Porto. Faculdade de Engenharia. 2011
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Ship-to-line na Bosch Car Multimedia Portugal, Ld.ªMeireles, Daniel José de Jesus Araújo January 2009 (has links)
Estágio realizado na Bosch Car Multimedia Portugal, Ld.ª e orientado pelo Eng.º Miguel Viana / Tese de mestrado integrado. Engenharia Industrial e Gestão. Faculdade de Engenharia. Universidade do Porto. 2009
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