Die Metastasierung in den Knochen ist beim fortgeschrittenen Mammakarzinom ein häufiges Problem. Insbesondere die Mammakarzinome, welche keinen Östrogenrezeptor α (ERα), keinen Östrogenrezeptor ß (ERß) und keine Überexpression des human epidermal growth factor receptor 2 (Her2/neu) aufweisen, die so genannten triple-negativen Mammakarzinome (TNBCs), werden als sehr aggressiv angesehen und haben eine schlechte Prognose. Kürzlich konnten wir zeigen, dass die Invasion von Mammakarzinomzellen in Kokultur mit der Osteoblastenähnlichen Zelllinie MG63 stark anstieg. Unter Verwendung dieses Modells haben wir nun untersucht, inwieweit der ERß eine Rolle bei der Zellinvasion von TNBC-Zellen in vitro spielt. Die ERα-und ERß-Proteinexpression wurde mithilfe des Western Blot-Verfahrens untersucht. Die Zellinvasion wurde mithilfe der Migrationsrate von TNBC-Zellen durch eine künstliche Basalmembran in einer modifizierten Boyden-Kammer während Kokultur mit der Osteoblastenähnlichen Zelllinie MG63 quantifiziert. Die Migration wurde mithilfe eines Scratch-Assays untersucht. Die Wirkung der ERß-Agonisten auf die CXC motif chemokine receptor 4 (CXCR4)-Proteinexpression während der Kokultur mit der Osteoblastenähnlichen Zelllinie MG63 wurde mithilfe des Western Blot-Verfahrens analysiert. Die Proliferation ist mit dem almarBlue®assay untersucht worden. Die TNBC-Zellen HCC1806 und HCC1937 zeigten keine ERα-Proteinexpression, jedoch eine hohe ERß-Proteinexpression. Die Zellinvasion der TNBC-Zelllinien HCC1806 und HCC 1937 zeigte in Kokultur mit Osteoblastenähnlichen Zelllinie MG63 einen signifikanten Anstieg. Die Behandlung mit den selektiven ERß-Agonisten Liquiritigenin und ERB-41 reduzierte die Invasion der TNBC-Zellen durch eine künstliche Basalmembran als Antwort auf eine zelluläre Stimulation. Während der Kokultur stieg die CXCR4-Proteinexpression der TNBC- Zelllinien HCC1806 und HCC1937 signifikant an. Die Behandlung mit Liquiritigenin resultierte in einem signifikanten Abfall der CXCR4-Proteinexpression. Beide ERß-Agonisten zeigten keinen Effekt auf die Proliferation der TNBC-Zelllinien. Unsere Ergebnisse weisen darauf hin, dass der ERß eine wesentliche Rolle bei der Invasion von TNBC-Zellen spielt. Die knochengerichtete Invasion kann durch ERß-Agonisten inhibiert werden.

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With the world's population growing at a rate faster than the rate at which natural resources are being replenished, the challenge for governments and the world's citizens is how to conserve resources in order to provide a sustainable level of natural resources for the future. Conserving natural resources includes educating the citizens of the world on the connection between natural resource depletion and their levels of consumption of resources, such as energy and water. To help alleviate the increasing burden the world's growing population is placing on natural resources, sustainability should be a part of college students' education in their field of study and in preparing them to become good citizens. This education should take place in the classroom and other activities, including athletics, community organizations, and in their residence life. Teaching students living in on-campus residence halls conservation activities provides information that students can use in their private lives when setting up their own households. On-campus residence halls also provide an opportunity to evaluate any gender differences related to conservation activities since the demographics of the residence halls vary from all-female, to co-ed, to all-male students. Virginia Polytechnic Institute and State University (Virginia Tech) was the location for a study on promoting environmentally-relevant behavior (ERB) among students residing in on-campus residence halls. The objective of this study was to investigate the relationship between the use of educational strategies and natural resource consumption by promoting ERB among students living in the residence halls during the spring and fall semesters of the 2009 calendar year. Using the literature on promoting ERB, five different strategies were designed for promoting water and electricity conservation. Each strategy involved different stimuli to promote student participation in ERB. The information provided the students included reasons why ERB was important and specific actions to take to conserve resources. In three of the strategies, students were provided the results of their conservation efforts monthly during the study period. The Virginia Tech Office of Residence Life provided detailed information for the 49 on-campus residence halls, including buildings' characteristics such as heating and cooling methods, age, construction, renovation history, square footage, if the buildings contained offices or classrooms, and student population figures. Variability among the buildings was eliminated by comparing these differences, and then a random numbers table was used to assign each of the buildings to one of the five different groups. The strategy for each group was applied to four residence halls -- two dormitories and two Greek Houses, for a total of twenty buildings. In each strategy more stimuli were applied in an effort to produce higher consumption reductions. The Virginia Tech Office of Facilities provided four-years historical electricity and seven-years historical water usage, and provided monthly usage for each building during the study period. Electricity consumption reduction was promoted in all twenty halls but water consumption reduction was promoted only in the dormitories, as the University was unable to track water consumption for any one individual Greek House. The historical data showed that water usage per student was higher in most of the female-occupied dormitories, but no statistical difference was seen with regards to historical electricity usage and gender. Percent change in per student usage — kilowatt hours for electricity and gallons for water — was the calculation used to determine change in ERB. The results of this research showed a general relationship between educational strategies and natural resource consumption reduction over both study periods. However, except for the Greek-House Spring semester results, no statistical significant difference was found between any of the different study groups. Electricity reductions were achieved in seventeen of twenty residence halls during the first semester and in all but one residence hall during the second semester. Water reductions were achieved in five of ten dormitories during the first semester and in six of nine dormitories in the second semester. However, the use of more strategies did not lead to a higher percentage of reductions. During the first semester, a statistically significant difference was found in water usage and gender and the difference did not support a female predisposition for ERB. Decreases were achieved in excess of 10% in the male-occupied dormitories, but only a minimal reduction or increases were achieved in any of the dormitories that included female residents. After the first month of the second semester, similar results were seen relative to gender, so additional posters and prompts were placed in the female-occupied dormitories. As a result, water reductions were achieved in six dormitories with only small increases in the other three, and the semester final results did not show a statistical significant difference between genders. The lack of statistical difference between the study groups could be a result of contamination, the active environmental organizations on campus, or an observational effect. The study was contaminated within the first two weeks of the study period when all residence halls across campus learned of the research and requested their inclusion in the study. Since, the residence halls in the control groups were advised of their inclusion in the study, the students may have demonstrated ERB because of the knowledge they were being observed. A survey sent to the students living in the study residence halls revealed that 94.6% of the students had knowledge of the study, and that 77% participated in ERB. Students showed a propensity for ERB when they were informed on their consumptive behaviors' effects on natural resource depletion, and by being provided with actions they could take to change their behaviors. This research did not show that adding strategies of feedback and group leaders to information increased the percentage of consumption reductions in college students residing on Virginia Tech's campus. / Master of Science

La première partie de mon travail de thèse a été de définir le rôle joué par le récepteur nucléaire Rev-erb alpha dans les mécanismes de synchronisation par la nourriture d’une horloge circadienne putative, non encore localisée, appelée « horloge alimentaire ». La seconde partie de mon travail a consisté à étudier la participation de Rev-erb alpha dans les régulations des métabolismes glucidique et lipidique. L’ensemble de nos données indique que le répresseur transcriptionnel Rev-erb alpha joue un rôle charnière dans les fonctions circadiennes ainsi que dans le métabolisme. En effet, d’un point de vue circadien, l’absence de Rev-erb alpha altère la synchronisation à l’heure des repas – démontré par une réduction des sorties comportementales et physiologiques de l’horloge alimentaire, ainsi que par l’absence d’ajustement du rythme de la protéine d’horloge PER2 dans l’oscillateur cérébelleux. Sur le plan métabolique, la délétion de ce gène modifie notamment le métabolisme des lipides – démontré par une accumulation excessive de tissu adipeux, une utilisation préférentielle des acides gras, ainsi qu’une perte de contrôle de l’expression de la Lipoprotéine lipase. / The work performed during this PhD thesis aimed at investigating the role of the transcriptional silencer Rev-erbα in both the circadian clockwork of the food-entrainable oscillator and metabolic regulations. Firstly, by evaluating food-anticipatory components in animals fed once a day at the same time, we showed that mice lacking Rev-erbα display a reduction in locomotor activity prior to food access compared to littermate controls. Accordingly, the rises in body temperature and corticosterone that anticipate mealtime are also diminished. Interestingly, daily p-ERK expression in hypothalamic regions and daily PER2 expression in the cerebellum of Rev-erbα KO mice are not phase-adjusted to feeding time. These results indicate that Rev-erbα participates in the integration of feeding signals and in food-seeking behaviors. Secondly, by investigating energy balance in fasted, normal chow or high-fat fed animals, we revealed that Rev-erbα KO mice exhibit greater reliance on lipid fuels as energy substrates, contributing to a mild hyperglycemic state. We also found that Lipoprotein lipase (Lpl) expression, is strongly up-regulated in peripheral tissues of Rev-erbα KO mice, predisposing mice to obesity. In this regard, we uncovered a new molecular pathway that ties clock-driven Lpl expression to energy homeostasis. These findings highlight the significance of daily Rev-erbα oscillations to prevent the appearance of the metabolic syndrome.In conclusion, we provide evidence that REV-ERBα may be a part of the food-entrainable oscillator clockwork that triggers food-anticipatory components, and represents a pivotal player to link the core clock machinery to metabolic pathways.

In this paper, we looked at how media influence education and how to provide students with tools for analyzing media messages in order for them to be able to reflect on that influence and foster critical thinking. We summarized research on media literacy in English language teaching and discussed how that may increase students’ language learning and critical thinking skills. To concretize our findings we developed a lesson plan using a contemporary YouTube genre, Epic Rap Battles of History, and used that as a basis for our discussions. This resulted in a discussion on the advantages of using media literacy to teach English and practical examples of how to do this in an English 6 course. We show the importance of connecting language learning to practical application as well as providing students with opportunities to develop their analytical and critical skills.

The nuclear receptor superfamily is responsible for regulating the expression of genes involved in development, reproduction and metabolism. These transcription factors control the expression of their target genes through the binding of small molecule regulators to their ligand binding domains. Classical nuclear receptors include the steroid receptors, which bind endocrine hormones and have been important targets of pharmaceutical intervention. However, approximately one half of the human nuclear receptors remain orphans and are without known cognate ligands. Focusing on the Drosophila orthologues of these orphan receptors, this project used mass spectrometry to identify the chemical diversity associated with the receptors following expression in recombinant systems. In a genome-wide screen of Drosophila nuclear receptors, this approach identified co-purifying molecules with a number of receptors. The physiological relevance of these putative ligand/receptor pairs was determined through biochemical analysis, in vivo characterization and structure determination. Ligand(s) or the ligand state was identified for the Drosophila receptors: DHR3, DHR96, E75, Ftz-f1 and USP. Of these, three were validated through the efforts of this project, and independent groups confirmed the remaining two. The most significant findings were the discoveries that the fly nuclear receptor E75 is regulated by heme, gas and redox, and that there is a similar regulatory scheme in the human orthologues, Reverbα and β. Furthermore, crystallization of the heme-bound Rev-erbβ ligand binding domain was also achieved, and this provided key insights into the mechanism of ligand regulation for the Rev-erbs. This project highlighted the role of nuclear receptors in metabolic surveillance. The ligands/signals identified in association with these receptors include: cholesterol, dehydrocholesterol, heme, NO, CO, redox and phospholipids. Unlike the classical steroid hormones, these are not dedicated signaling molecules, but instead are key substrates or products of metabolism. In the context of nuclear receptor signaling, I hypothesize that these metabolites serve as metabolic indicators/signals in the regulation of development and metabolism. Furthermore, four of these Drosophila receptors comprise the ecdysone-response pathway in the developing fly. Taken together, this suggests that both the metabolic state of the organism and steroid hormones drive nuclear receptor regulation of development.

The nuclear receptor superfamily is responsible for regulating the expression of genes involved in development, reproduction and metabolism. These transcription factors control the expression of their target genes through the binding of small molecule regulators to their ligand binding domains. Classical nuclear receptors include the steroid receptors, which bind endocrine hormones and have been important targets of pharmaceutical intervention. However, approximately one half of the human nuclear receptors remain orphans and are without known cognate ligands. Focusing on the Drosophila orthologues of these orphan receptors, this project used mass spectrometry to identify the chemical diversity associated with the receptors following expression in recombinant systems. In a genome-wide screen of Drosophila nuclear receptors, this approach identified co-purifying molecules with a number of receptors. The physiological relevance of these putative ligand/receptor pairs was determined through biochemical analysis, in vivo characterization and structure determination. Ligand(s) or the ligand state was identified for the Drosophila receptors: DHR3, DHR96, E75, Ftz-f1 and USP. Of these, three were validated through the efforts of this project, and independent groups confirmed the remaining two. The most significant findings were the discoveries that the fly nuclear receptor E75 is regulated by heme, gas and redox, and that there is a similar regulatory scheme in the human orthologues, Reverbα and β. Furthermore, crystallization of the heme-bound Rev-erbβ ligand binding domain was also achieved, and this provided key insights into the mechanism of ligand regulation for the Rev-erbs. This project highlighted the role of nuclear receptors in metabolic surveillance. The ligands/signals identified in association with these receptors include: cholesterol, dehydrocholesterol, heme, NO, CO, redox and phospholipids. Unlike the classical steroid hormones, these are not dedicated signaling molecules, but instead are key substrates or products of metabolism. In the context of nuclear receptor signaling, I hypothesize that these metabolites serve as metabolic indicators/signals in the regulation of development and metabolism. Furthermore, four of these Drosophila receptors comprise the ecdysone-response pathway in the developing fly. Taken together, this suggests that both the metabolic state of the organism and steroid hormones drive nuclear receptor regulation of development.

No description available.

Markéta Hrdličková, Ennoblement and it's process in Czech lands during the reign of emperor Leopold I., based on acts of Bohemian Court Chancellery Abstract This thesis is based on the research of files of Bohemian Court Chancellery during the reign of emperor Leopold I. (1657-1705). It resolves about the emperor Leopold and the events and affairs of his time, and those which preceded it and had a significant influence on the reasonings mentioned in the files and acts of ennoblement. It also contains chapters about nobility of the Lands of the Bohemian Crown, the process of ennoblements and its files. Addendum contains a list of ennoblement acts in alphabetical order regarding both persons and families.

INTRODUÇÃO: Os astrocitomas difusamente infiltrativos são os tumores mais freqüentes de Sistema Nervoso Central (SNC) com uma taxa de 5-7 novos casos por 100.000 pessoas ano. São tumores altamente invasivos e estão associados com alterações de alguns genes como EGF (fator de crescimento epidérmico) e o EGFR (receptor do fator de crescimento epidérmico), que podem criar um aumento da atividade mitogênica, acarretando aumento de proliferação e maturação celular, apoptose, angiogênese e metástase. O nível de expressão destes genes pode ser influenciado por alterações genéticas, como a presença de polimorfismos. Uma mudança única de base (SNP) pode alterar a expressão gênica e, sendo assim, estar associada ao aumento do risco de desenvolver astrocitomas. Nesse trabalho, foram analisados 2 SNPs na região não traduzida (c.-191C>A e c.-216G>T) e um SNP no exon 16 (c.2073A>T) do gene EGFR, e um outro SNP na região não traduzida no gene EGF (c.61A>G). Os SNPs foram associados a expressão gênica do EGFR e a sobrevida dos pacientes. MÈTODOS: Foi realizado um estudo caso-controle com 193 casos de astrocitomas difusamente infiltrativos e 200 controles por amplificação por PCR seguido de digestão enzimática. Os produtos digeridos das amostras foram analisados por eletroforese em gel de agarose e poliacrilamida e corados com brometo de etídeo. A expressão gênica foi realizada após extração de RNA do tecido tumoral seguida de transcrição reversa e PCR em tempo real. Testes de qui-quadrado, odds ratio (OR), intervalo de confiança 95% (IC95%), t de Student e curvas de Kaplan-Meier foram realizados para análises estatística. RESULTADOS: A análise das freqüências dos genótipos dos polimorfismos mostrou uma diferença na distribuição entre casos e controles para o polimorfismo c.2073A>T. Pacientes com o genótipo TT apresentou um menor risco para astrocitoma quando comparados com o genótipo AA (OR=0,51, IC95%=0,29-0,99). Nenhuma correlação foi encontrada para os outros polimorfismos analisados. Também não foi encontrada correlação entre os genótipos dos polimorfismos e os níveis de expressão de EGFR e a sobrevida dos pacientes. CONCLUSÃO: Nosso trabalho mostrou haver um possível fator de proteção quando o paciente é portador do genótipo TT, o que pode levar a uma diminuição do risco de desenvolver o tumor. Pacientes com genótipo TT do polimorfismo c.2073A>T do gene EGFR apresentam um menor risco para astrocitomas difusamente infiltrativos do que os com o genótipo AA. / INTRODUCTION: Diffusely infiltrative astrocytomas are the most frequent tumors of the Central Nervous System (CNS) with a rate of 5-7 new cases in 100,000 individuals per year. They are highly invasive, and they are associated to alterations in some genes as EGF (epidermal growth factor) and EGFR (epidermal growth factor receptor), which may increase mitogenic activity, leading to increase of proliferation, cellular maturation, apoptosis, angiogenesis, and metastasis. Genetic alterations, as presence of polymorphisms of single nucleotide change (SNP) could influence their expression level, and thus could be associated to increased risk in developing astrocytomas. In the present study, two SNP of non-coding region (c.-191C>A and c.-216G>T) and one SNP in exon 16 (c.2073A>T) of EGFR, and another SNP of non-coding region of EGF (c.61A>G) were analyzed. The SNPs were associated to EGFR expression level and to survival time. METHOD: a case-control study of 193 of diffusely infiltrative astrocytomas and 200 controls was carried out, with PCR amplification and enzymatic digestion, which products were analyzed in agarose gel or polyacrylamide gel electrophoresis stained by ethidium bromide. EGFR expression level was studied by real time PCR after RNA extraction followed by reverse transcription of tumor tissues compared to epileptic non-neoplastic brain tissues. Stastistical analysis were performed by chi-square, odds ratio (OR), 95% confidence interval (95% CI), Student-t test and Kaplan Meier. RESULTS: The polymorphic genotype frequency was different between case and controls for the polymorphism c.2073A>T. Patients with TT genotype presented lower risk to develop astrocytoma when compared to genotype AA (OR=0.51, CI95%=0.29- 0.99). No other correlation was observed for the remaining studied polymorphisms. There was neither correlation between the polymorphic genotypes and the EGFR expression levels nor with survival time. CONCLUSION: The present study showed a possible protection factor in developing astrocytomas for the patients harboring the genotype TT of c.2073A>T polymorphism of EFGR, thus the patients presenting TT genotype have lower risk to develop diffusely infiltrative astrocytoma than patients presenting the genotype AA.