Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-03-07T11:12:39Z No. of bitstreams: 1 andersonchristianpereira.pdf: 1068847 bytes, checksum: bb25159eb6b54e6585618bb0d399b65d (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-04-24T01:57:10Z (GMT) No. of bitstreams: 1 andersonchristianpereira.pdf: 1068847 bytes, checksum: bb25159eb6b54e6585618bb0d399b65d (MD5) / Made available in DSpace on 2016-04-24T01:57:10Z (GMT). No. of bitstreams: 1 andersonchristianpereira.pdf: 1068847 bytes, checksum: bb25159eb6b54e6585618bb0d399b65d (MD5) Previous issue date: 2013-10-31 / Este estudo se dedicou a analisar como ocorreu a implementação da política pública para o Ensino Médio, vigente no Estado de Minas Gerais, em três escolas estaduais pertencentes à circunscrição da Superintendência Regional de Ensino de Passos (SRE de Passos), observando a forma com que essas normatizações foram difundidas nos estabelecimentos de ensino para, então, propor um Plano de Ação Educacional que possibilite a melhoria na assertividade desse processo de implementação, que apresentou várias falhas na comunicação institucional. A temática é relevante, uma vez que essa etapa da Educação Básica é problemática no país todo, detentora dos piores índices de proficiência, e muitas têm sido as reformas propostas para reverter a situação. Participaram da pesquisa de campo profissionais das escolas e da SRE de Passos, dos quais foram colhidos depoimentos. Também são considerados os dados apurados pelo CAEd, Inep, Meritt e SRE de Passos, bem como a legislação vigente, tanto estadual como federal. Os referenciais bibliográficos utilizados são, principalmente, de autoria de Claudio de Moura Castro, Simon Schwartsman, Maria Helena Guimarães de Castro, Sergio Tiezzi, Idalberto Chiavenato, Gustavo Gomes de Matos, Henry Mintzberg e Heloísa Lück, entre outros. Ao final das análises, que revelaram a existência de problemas na comunicação institucional desse processo de implementação, é formulado e apresentado um Plano de Ação Educacional, voltado à Gestão Escolar, no qual se propõem ações que venham a contribuir com a melhoria da comunicação institucional durante o processo de implementação de políticas públicas educacionais, sendo este uma adaptação do plano elaborado por Gustavo Gomes de Matos, em sua “Comunicação sem Complicação”. / This study was devoted to analyze how was performed the implementation of public policy for the High School, effective in the State of Minas Gerais, in three state schools belonging to the constituency of the Superintendência Regional de Ensino de Passos (SRE de Passos), observing the way that these norms were widespread in schools, to then propose an Educational Action Plan enabling improvement of assertiveness in this implementation process, that presented several deficiencies in the institutional communication. The theme is relevant, since this stage of Basic Education is problematic throughout the country, holder of the worst rates of proficiency, and many have been proposed reforms to reverse the situation. Professionals from schools and SRE de Passos have participated in the field research, which have given testimonies. Data compiled by CAEd, Inep, Meritt and SRE de Passos were also considered, as well as state and federal legislation. The bibliographical references are used, mainly, authored by Claudio de Moura Castro, Simon Schwartsman, Maria Helena Guimarães de Castro, Sergio Tiezzi, Idalberto Chiavenato, Gustavo Gomes de Matos, Henry Mintzberg and Heloísa Lück, among others. At the end of the analysis, which revealed the existence of problems in institutional communication in this implementation process, an Educational Action Plan has been formulated and submitted, returned to school management, which proposes actions that will contribute to the improvement of institutional communication during the process of implementing education public policy, which is an adaptation of that developed by Gustavo Gomes de Matos, in his "Communication without Entanglement".

Cette étude s’inscrit dans le domaine de la microélectronique. Elle consiste à étudier l’impact de la morphologie 3D du dépôt d’énergie sur la modélisation des événements singuliers (SEE) induits par les muons atmosphériques. La démarche a consisté dans un premier temps à modéliser le dépôt d’énergie induits par des protons dans des volumes nanométriques. Pour cela, le code Monte-Carlo GEANT4 a permis de simuler et de stocker dans une base de données, les caractéristiques des traces des dépôts d’énergie des protons. Une fois la démarche validée pour les protons, des simulations du dépôt d’énergie induit par des muons ont été réalisées. Une caméra CCD a été utilisée afin de réaliser des mesures de l’environnement radiatif atmosphérique et de contraindre la modélisation des dépôts d’énergie induits par les muons. Cette étude met en évidence et quantifie l’apport de considérer la distribution radiale du dépôt d’énergie induit par des protons pour des volumes nanométriques, dans les calculs de prédiction des SEE. En revanche, l’étude montre que la considération de la distribution radiale du dépôt d’énergie induit par des muons dans des volumes nanométriques a un impact négligeable sur la modélisation des SEE. Il serait intéressant de réaliser des mesures du dépôt d’énergie induit par des muons dans des technologies nanométriques sous accélérateur. Cela permettrait d’apporter des données expérimentales encore inexistantes, nécessaires au développement de nouveaux modèles physiques plus précis sur la modélisationdu dépôt d’énergie induit par des muons. / This study concerns the domain of the microelectronics. It consists in the studyof the impact of the 3D morphology of the energy deposit on the Single EventEffect (SEE) modeling, induced by atmospheric muons. Over a first phase, theapproach has consisted in the modeling of the energy deposit induced by protonsin nanometric volumes. For that purpose the use of the Monte Carlo code GEANT4has allowed to simulate and stock in a database the tracks characteristics of theenergy deposit induced by protons. Once the approach validated for the protons,simulations of the energy deposit induced by muons have been realized. A CCDcamera has been used in order to measure the radiative atmospheric environmentand to constrain the modeling of the energy deposit induced by muons. This studyhighlights and quantify the contribution of the radial distribution of the energydeposit incuced by protons in nanometric volumes for the SEE prediction. On theother hand, the study shows that the contribution of the radial distribution of theenergy deposit induced by muons in nanometric volumes has a negligeable impacton the SEE modeling. It will be interesting to realize measurements of the energydeposit induced by muons in nanometric technologies under accelerator. This willallow to bring experimental data still nonexistant necessary to the developpmentof new physical models more accurate on the modelization of the energy depositinduced by muons.

Sediments of Bohemian Forest lakes are important natural archives. Their sedimentary record covers postglacial history of the lakes as well as history of natural processes in a wider region. It also documents local settlements and changes in landscape management. The lake sediments have attracted the interest of the scientific community since the end of the 19th century. Despite of the fact that modern paleolimnological and paleoecological investigations were already performed in the second half of the 20th century in the Bohemian Forest Mts., the great potential of the lake sediments was not fully utilized in scientific research so far. The ultimate objective of this thesis is to deepen knowledge of these natural archives and support their utilization in future studies. Several specific objectives have been set to achieve the ultimate objective: i) to compare age of the Bohemian Forest lake sediments with the recent knowledge of local deglaciation at the end of the last ice age, ii) to integrate chronostratigraphic marker horizons as one of the tools of Late Glacial sediment dating, iii) to assess the role of bioerosion in chitinous subfossil freshwater invertebrate remains on the record representativeness, and iv) to prove the presumed dystrophic nature of the lakes during the Holocene using...

When a cell divides into two, it is imperative that each resultant daughter receives a full complement of chromosomes; DNA is ultimately responsible for all cellular processes. Cohesion between sister chromatids from the moment of their generation in S phase is central to ensuring the fidelity of chromosome segregation. Smc1 and Smc3 proteins interact with each other via their hinges and with a bridging kleisin subunit via their heads to form the cohesin ring. It is cohesin, through entrapment of sister chromatid within its ring, that confers sister chromatid cohesion. The process of cohesin’s loading onto DNA is poorly understood. While it is thought to depend on ATP hydrolysis, opening of the ring at one of its three interfaces, and the as yet undefined action of the kollerin complex, comprising Scc2 and Scc4 proteins, the sequence of events as they occur are yet to be defined. A recent screen for suppressors of a thermosensitive scc4 allele in budding yeast revealed a mutation within Smc1’s hinge that could bypass the kollerin subunit. Here, the Smc1 suppressor mutation is investigated. Through targeted mutagenesis, the Smc1D588Y mutant identified in the screen and two additional point mutants, Smc1D588F and Smc1D588W, are herein proven able to bypass Scc4 function completely. Thus we provide the strongest evidence to date to suggest that cohesin’s hinge is a critical factor in its loading. Biochemical evidence shows that isolated Smc1 hinge mutants are defective in their binding to Smc3 hinges. This, together with the genetic link made between the hinge and loading complex, suggests that hinge opening might be a requisite for loading. Through mutagenesis of Scc2 and Scc4 we show that the N-terminus of each protein is responsible for their dimerisation. Furthermore, the N- terminus of Scc2 confers no function other than in its binding to Scc4. Finally, we show that Scc4 is required for the enrichment of both Scc2 and cohesin at centromeres, but not at arm loci. Our results are therefore indicative of there being two different pathways of cohesin loading.

Subcellular sequestration of proteins within membrane bound compartments is widely acknowledged to be an important mode of enzymatic regulation. Recently a novel paradigm for metabolic enzyme compartmentalisation has become apparent with the identification of several proteins which are able to form filamentous structures in vivo. Multiple studies independently identified the essential de novo pyrimidine biosynthesis enzyme CTP synthetase as a major constituent of a novel filamentous structure which has been termed “the cytoophidium”. Cytoophidia have been observed to form in multiple organisms including bacteria (C. crescentus), yeast (S. cerevisiae) and fruit fly (D. melanogaster) as well as in human cultured cells. In this thesis I describe the development and results of a high throughput genomescale screen to identify factors involved in cytoophidia biogenesis. Observations of tissue specific CTPS distribution lead to the identification of the well-conserved growth regulator dm/dMyc as an essential factor for CTPS regulation in vivo. These results provide new insights into the coordination of cellular growth and metabolic regulation during normal development and indicate the potential of CTPS/cytoophidia as a future therapeutic target.

Phylogenetic studies are providing powerful new insights into the evolution of complex traits. Metal hyperaccumulation is an unusual and complex physiological trait found in about 500 plant species and is associated with an exceptionally high degree of tolerance of metalliferous soils. Alyssum L. (Brassicaceae) is the largest known hyperaccumulator genus, comprising approximately 188 species distributed throughout the Mediterranean region and south-west Asia. Approximately one-quarter of these are largely restricted to areas of serpentine soils and have the ability to accumulate nickel to high concentrations in shoot tissue. This genus provides a good example in which to study the origins of a complex physiological trait, but its phylogeny is currently poorly understood. To produce a well-resolved phylogenetic tree to investigate the number and timing of origins of nickel hyperaccumulation within Alyssum, DNA sequences were generated for four chloroplast regions (matK, rps16–trnK, trnD–T and trnL–F) from 170 of 255 species in the tribe Alysseae. Additional sequencing was carried out for the chloroplast genes ndhF and rbcL and the nuclear gene PHYA. A Bayesian analysis employing a relaxed uncorrelated lognormal molecular clock and multiple fossil-age calibration points was carried out to reconstruct a time-calibrated phylogeny of this tribe using appropriate outgroups. Optimization of the nickel hyperaccumulation trait onto the resulting phylogenetic tree suggests that nickel hyperaccumulation arose twice in the Alysseae in the late Miocene/early Pliocene: 3.3–8.3 Mya in Alyssum and 6.3–8.8 Mya in Bornmuellera. The single origin in Alyssum is strongly associated with a significant acceleration in net species diversification rate, suggesting the ability to hyperaccumulate nickel could have provided a key evolutionary innovation facilitating rapid range expansion and subsequent species diversification. The scattered distribution of nickel hyperaccumulators across small island-like patches of serpentine soil suggests that allopatric speciation may have driven rapid diversification in this clade.

This thesis addresses the methodological challenges of determining the variability of large herbivore populations through time and their impact on European vegetation. Large herbivores are at the heart of conservation policy. However, opinions widely diverge on whether we should aim for fewer herbivores and managed populations or, on the contrary as advocated by the rewilding movement, more herbivores and self-regulating populations acting as ecosystem engineers. This controversy has roots in a debate regarding the nature of ecosystems before the prevalence of human activities. Baseline ecosystems are either described as continuous forest cover with passive large herbivores, or, in contrast, as mosaics with patchy forest cover driven <i>inter alia</i> by bison, aurochs and horses, now rare or extinct in Europe. The main obstacle in moving this debate forward is a poor understanding of large-herbivore densities in the past. I analysed modern pollen and spore assemblages from known environmental settings to improve palaeoecological interpretation of fossil assemblages dating from the pre-human (baseline) period. The sites investigated are the rewilded grasslands of the Oostvaardersplassen (The Netherlands), the mosaic habitats of The New Forest (UK) and the old-growth closedcanopy forest of Białowieża (Poland). I demonstrate that the common practice of interpreting pollen percentages fails to estimate past forest cover in situations with natural grazing. As an explanation, I suggest that pollen productivity fluctuates with biotic factors such as herbivory and canopy shading. As a result, new insights into the baseline debate require additional lines of evidence. In this thesis, I develop an existing methodology to reconstruct past herbivore presence using fossil dung fungal spores. I synthesise current knowledge of this method with an emphasis on spore identification and, finally, I demonstrate that dung fungal spore abundance in lake sediments can be translated into large herbivore numbers. The evidence presented in this thesis contributes to the debate on re-wilding and addresses a fundamental challenge of nature conservation in the human-dominated landscapes of Europe.

Endoplasmic reticulum (ER)-associated degradation (ERAD) of misfolded proteins of the secretory pathway is crucial for ER homeostasis and the physiological importance of this mechanism is reflected by more than 60 diseases that have been linked to ERAD to date. The best characterised mammalian ERAD complex is centred on the ubiquitin ligase Hrd1, and for a complete understanding of the dynamics of the ERAD network it is important to thoroughly characterise the interactions within the Hrd1 complex and to decipher the functions of the individual accessory factors. SEL1L is a well characterised interaction partner of Hrd1 and here we identify a highly hydrophobic region in the lumenal part of SEL1L as necessary, but not sufficient, to interact with Hrd1; as a consequence, the topology of SEL1L may need to be re-evaluated. Furthermore, we investigate the roles of the novel Hrd1 interaction partners AUP1 and FAM8A1 in ERAD. We establish here that AUP1 adds to the complexity of the ERAD network by making the cognate E2 of gp78, Ube2g2, accessible to Hrd1 and by regulating the access of polyubiquitinated proteins to the Hrd1 complex. Moreover, we demonstrate that Hrd1 is the regulator of FAM8A1 half-life and that the interaction between these two proteins is necessary for degradation of the lumenal ERAD model substrate TTR_D18G.

Membrane-bound vesicle secretion provides a novel intercellular communication mechanism, whose roles and regulation remain poorly characterised, particularly in vivo. I have identified two classes of lipid-containing, vesicle-like structures secreted into seminal fluid by epithelial cells of the Drosophila male accessory gland (AG). Exosomes, one class of membrane-bound vesicle formed inside late endosomal multivesicular bodies, are specifically secreted by secondary cells (SCs). The unusual cell biology of SCs allowed me to develop a powerful new high resolution in vivo system to characterise the mechanisms underlying intracellular membrane trafficking events underlying exosome biogenesis using real-time live imaging. I characterise how specific ESCRTs (endosomal sorting complexes required for transport) control SC exosome biogenesis, and identify a novel role for BMP signalling in regulating endolysosomal trafficking events necessary for exosome secretion. I also identify roles for epidermal growth factor receptor (EGFR) and phosphatidylinositide 3-kinase (PI3K) signalling in exosome biogenesis. Importantly, SC exosomes are transferred to females during mating. Here, they fuse with sperm, mirroring in vitro interactions between human prostate exosomes and sperm, and interact with the female reproductive tract epithelium. Blocking SC exosome production specifically suppresses post-mating effects on female receptivity to remating, demonstrating that exosomes have an important reproductive signalling function in vivo, directly or indirectly reprogramming female cells. Finally, I show that main cells, the major epithelial AG cell type, shed lipid-containing microvesicle-like structures from their apical surface. Remarkably, these vesicles carry the seminal peptide, sex peptide, into females during mating and also contribute to the anterior mating plug. In summary, my data reveal previously unsuspected roles for exosomes and microvesicles in Drosophila reproduction that may be evolutionarily conserved. Since these vesicles mediate physiological processes previously thought to involve soluble peptides, my work suggests that current models explaining male reprogramming of female behaviours in flies and higher organisms need substantial revision.

Evidence in different systems suggests that local translation is involved in synaptic plasticity in both neuron and muscle, but the mechanism by which this occurs is still poorly understood. The mRNA-binding protein Syncrip is conserved from fly to mammals and is thought to be involved in localized translation in both oocytes and neurons. Previous work has shown that Syncrip associates with mRNAs encoding key synaptic proteins at the Drosophila larval neuromuscular junction. Here I show that Syncrip is necessary for the structure and function of the neuromuscular junction. First, the loss of Syncrip leads to overgrowth of the neuromuscular junction. Second, Syncrip is required for proper expression of the Ca²⁺-sensor Synaptotagmin1 at the presynapse, and loss of Syncrip causes a decrease in vesicle release probability. Third, while it was not possible to measure mRNA distribution in neurons, Syncrip mutants, like other perturbations in synaptic plasticity, correlate with changes in mRNA localization in muscle. Fourth, the overexpression and loss of Syncrip function suggest that the nuclear and nucleolar trafficking of the eukaryotic translation initiation factor eIF4E may be important to regulating synaptic morphology. These data suggest that Syncrip is involved in mRNA localization and translation in synaptic plasticity.