<p>Cyclooxygenase-2 (COX-2) catalyses prostaglandin synthesis from arachidonic acid during inflammation. COX-2 is expressed in the normal brain and is induced in neurological disorders. There is evidence that COX-2 is involved in secondary events leading to cell death in the brain. The first objective was to study the expression of COX-2 in the brain after kainate (KA)-induced limbic seizures and brain trauma caused by controlled cortical contusion (CCC) and fluid percussion injury (FPI). COX-2 mRNA and protein were strongly induced by limbic seizures in the hippocampus, amygdala and piriform cortex. CCC and FPI resulted in an upregulation of COX-2 mainly in the dentate gyrus and cortex, with differences in expression levels in these regions between the models. The second objective was to evaluate the effects of selective COX-2 inhibitors on delayed cell death. Limbic seizures induced cell death in parts of the hippocampus, amygdala and functionally connected regions. Treatment with the selective COX-2 inhibitor rofecoxib 8 h after KA injection significantly reduced hippocampal cell death. Pre-treatment with the COX-2 inhibitor nimesulide augmented acute seizures with increased mortality and thus the effect of nimesulide on delayed cell death could not be evaluated. Effects of rofecoxib on trauma-induced cell death were studied in the FPI model. FPI induced delayed cell death mainly in the ipsilateral cortex and bilaterally in the dentate gyrus. Rofecoxib treatment, starting directly after injury was caused, had no protective effect against cell death. </p><p>The results suggest that COX-2 inhibition may be both detrimental and beneficial and largely dependent on the time schedule of treatment. COX-2 inhibitors might thus be of value as a neuroprotective treatment approach, provided that the role of COX-2 and the time course of effects of its metabolites in the brain are elucidated.</p>

Cyclooxygenase-2 (COX-2) catalyses prostaglandin synthesis from arachidonic acid during inflammation. COX-2 is expressed in the normal brain and is induced in neurological disorders. There is evidence that COX-2 is involved in secondary events leading to cell death in the brain. The first objective was to study the expression of COX-2 in the brain after kainate (KA)-induced limbic seizures and brain trauma caused by controlled cortical contusion (CCC) and fluid percussion injury (FPI). COX-2 mRNA and protein were strongly induced by limbic seizures in the hippocampus, amygdala and piriform cortex. CCC and FPI resulted in an upregulation of COX-2 mainly in the dentate gyrus and cortex, with differences in expression levels in these regions between the models. The second objective was to evaluate the effects of selective COX-2 inhibitors on delayed cell death. Limbic seizures induced cell death in parts of the hippocampus, amygdala and functionally connected regions. Treatment with the selective COX-2 inhibitor rofecoxib 8 h after KA injection significantly reduced hippocampal cell death. Pre-treatment with the COX-2 inhibitor nimesulide augmented acute seizures with increased mortality and thus the effect of nimesulide on delayed cell death could not be evaluated. Effects of rofecoxib on trauma-induced cell death were studied in the FPI model. FPI induced delayed cell death mainly in the ipsilateral cortex and bilaterally in the dentate gyrus. Rofecoxib treatment, starting directly after injury was caused, had no protective effect against cell death. The results suggest that COX-2 inhibition may be both detrimental and beneficial and largely dependent on the time schedule of treatment. COX-2 inhibitors might thus be of value as a neuroprotective treatment approach, provided that the role of COX-2 and the time course of effects of its metabolites in the brain are elucidated.

Concerns about climate changes and scarcity of fossil fuels are rising. Hence wood is becoming an attractive source of renewable energy and raw material and these new dimensions have prompted increasing interest in wood formation in trees, in both the scientific community and wider public. In this thesis, the focus is on a key process in wood development: programmed cell death (PCD) in the development of xylem elements. Since secondary cell wall formation is dependent, inter alia, upon the life time of xylem elements, the qualitative features of wood will be affected by PCD in xylem, about which there is little information. This thesis focuses on the anatomical, morphological and transcriptional features of PCD during xylem development in both the stem of hybrid aspen, Populus tremula (L.) x tremuloides (Michx.) and the hypocotyl of the herbaceous model system Arabidopsis thaliana (L. Heynh.). In Populus, the progressive removal of organelles from the cytoplasm before the time of death (vacuolar bursts) and the slowness of the cell death process, illustrated by DNA fragmentation assays (such as TUNEL and Comet assays), have been ascertained in the xylem fibres by microscopic analyses. Furthermore, candidate genes for the regulation of fibre cell death were identified either from a Populus EST library obtained from woody tissues undergoing fibre cell death or from microarray experiments in Populus stem, and further assessed in an in silico comparative transcriptomic analysis of Arabidopsis thaliana. These candidate genes were either putative novel regulators of fibre cell death or members of previously described families of cell death-related genes, such as autophagy-related genes. The induction of the latter and the previous microscopic observations suggest the importance of autophagy in the degradation of the cytoplasmic contents specifically in the xylem fibres. Vacuolar bursts in the vessels were the only previously described triggers of PCD in the xylem, which induce the very rapid degradation of the nuclei and surrounding cytoplasmic contents, therefore unravelling a unique previously unrecorded type of PCD in the xylem fibres, principally involving autophagy. Arabidopsis is an attractive alternative model plant for exploring some aspects of wood formation, such as the characterisation of negative regulators of PCD. Therefore, the anatomy of Arabidopsis hypocotyls was also investigated and the ACAULIS5 (ACL5) gene, encoding an enzyme involved in polyamine biosynthesis, was identified as a key regulator of xylem specification, specifically in the vessel elements, though its negative effect on the cell death process. Taken together, PCD in xylem development seems to be a highly specific process, involving unique cell death morphology and molecular machinery. In addition, the technical challenges posed by the complexity of the woody tissues examined highlighted the need for specific methods for assessing PCD and related phenomena in wood. / Oron för klimatförändringar och brist på fossila bränslen har ökat påtagligt under de senaste åren. De enorma möjligheter som skogsråvaran erbjuder som alternativ källa för förnyelsebar energi och råmaterial har väckt ett stort intresse också för den biologiska processen bakom vedbildning i träd. Denna avhandling fokuserar på en viktig process i vedbildning: programmerad celldöd (PCD) i xylemet. Xylemcellernas livstid påverkar bildningen av sekundära cellväggar, vilket i sin tur påverkar vedens kvalitativa egenskaperna, så som veddensitet. Trots dess betydelse för viktiga egenskaper hos vedråvaran existerar fortfarande väldigt lite information om xylem PCD på cellulär eller molekylär nivå. I den här avhandlingen belyses de anatomiska, morfologiska och genetiska aspekterna av PCD i xylemutveckling i både stam av hybridasp, Populus tremula (L.) x tremuloides (Michx.) och hypokotyl av det örtartade modellsystemet Arabidopsis thaliana (L. Heynh.). Xylemet i både Populus och Arabidopsis består av två olika celltyper; de vattentransporterade kärlen och de stödjande fibrerna. Det är känt att celldöd i kärlen pågår mycket snabbt efter att den centrala vakuolen brister och de hydrolytiska enzymer släpps in i cytoplasman. I den här avhandlingen ligger fokus på fibrerna i Populus xylemet. Med hjälp av mikroskopianalyser av cellmorfologin (elektronmikroskopi) och DNA-fragmentering i cellkärnan (TUNEL- och Comet-analyser) kunde vi konstatera att till skillnad från kärlen så uppvisar fibrerna en långsam och progressiv nedbrytning av organellerna och cellkärnans DNA före vakuolbristning. Dessutom har kandidatgener för reglering av fibercelldöd identifierats antingen från ett Populus EST bibliotek från vedartade vävnader som genomgår fibercelldöd eller från mikroarray experiment i Populus stam. Dessa kandidatgener är antingen potentiella nya regulatorer av fibercelldöd eller medlemmar av tidigare beskrivna familjer av celldödsrelaterade gener. Bland de sistnämnda finns autofagi-relaterade gener, vilket stöder funktionen av autofagi i samband med autolys av cellinnehållet i xylemfibrerna. Dessa studier pekar därför på en typ av PCD som har inte tidigare beskrivits för xylemet. Arabidopsis är ett alternativt växtmodellsystem för studier av vissa aspekter av vedbildningen, såsom karakteriseringen av negativa regulatorer av PCD. Därför har också hypokotylanatomin analyserats, och ACAULIS5 (ACL5) genen, som kodar för ett enzym i biosyntesen av polyaminer, har visats vara en viktig regulator av xylemspecifikation genom dess negativa effekt på kärlens celldöd. Sammantaget visar denna avhandling att PCD i xylemutvecklingen verkar involvera unika morfologiska och molekylära mekanismer. Vi visar dessutom att komplexiteten hos de vedartade vävnaderna leder till ett behov av bättre anpassade verktyg för att djupare kunna bedöma PCD och liknande fenomen i veden. / Även med namnet Moreau-Courtois, Charleen L. samt Moreau, Charleen.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-07-24T13:25:47Z No. of bitstreams: 1 julianapolisseni.pdf: 208270479 bytes, checksum: 5c1499cab892f30dc1d6d2d17268e467 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-08-09T13:28:51Z (GMT) No. of bitstreams: 1 julianapolisseni.pdf: 208270479 bytes, checksum: 5c1499cab892f30dc1d6d2d17268e467 (MD5) / Made available in DSpace on 2017-08-09T13:28:51Z (GMT). No. of bitstreams: 1 julianapolisseni.pdf: 208270479 bytes, checksum: 5c1499cab892f30dc1d6d2d17268e467 (MD5) Previous issue date: 2013-05-14 / FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais / Objetivou-se avaliar sistemas de cultivo propostos para utilização de embriões humanos: sistema de cultivo único e sistema de cultivo sequencial, comparando com sistema de cultivo utilizado nos modelos experimentais bovino e ratas Wistar. Especificamente avaliou-se taxa de clivagem e blastocisto, cinética do desenvolvimento embrionário, quantificação da expressão dos genes em blastocistos bovinos e avaliação da viabilidade e qualidade por morfologia e Tunel de embriões oriundos de ratas Wistar. Para bovinos, complexos cumulus-oócitos foram maturados, fecundados in vitro. No experimento 1 os zigotos foram distribuídos nos sistemas de cultivo: CR2aa, SOFaa, sistema de cultivo seqüencial ECM®/Multiblast® e sistema de cultivo único Global®. Avaliou-se a taxa de clivagem e de blastocisto e expressão dos genes PRDX, HSP70.1, GLUT1 e GLUT5, nos blastocistos. No experimento 2, o cultivo embrionário foi avaliado sobre o desenvolvimento de embriões bovinos biopsiados. Os zigotos foram distribuídos nos sistemas de cultivo: CR2aa, sistema de cultivo seqüencial G1®/G2® e ECM®/Multiblast®, sistema de cultivo único Global®. Para todos os grupos, os embriões 8-16 células foram biopsiados e retornados para cultivo. Avaliou-se taxa de clivagem e de blastocisto. Para ratas Wistar, utilizou-se animais, do Centro de Biologia da Reprodução (CBR/UFJF), com doze semanas de vida, superovuladas com 150 Ul/kg de PMSG e 75 Ul/kg de hCG, intraperitonealmente. No experimento 1 a coleta embrionária procedeu-se 24 horas após o hCG e avaliou-se a viabilidade embrionária No experimento 2 ratas foram distribuídas entre os grupos controle e superovulado e os embriões foram coletados 48 e 72 h após a administração de hCG. Peso dos ovários, número total de estruturas embrionárias e grau de qualidade de embriões foram analisados. No experimento 3 a coleta embrionária procedeu-se 72 h após a administração de hCG. Avaliou-se a qualidade de embriões cultivados in vitro oriundos de ratas superovuladas através da taxa de blastocisto e do índice apoptótico. Já no experimento 4 a coleta dos embriões foi realizada 48h após a administração do hCG. Embriões recuperados foram distribuídos aleatoriamente segundo os grupos: sistema de cultivo sequencial ECM°/Multiblast®, sistema de cultivo único Global®, KSOM. Avaliou-se taxa de blastocisto. A análise estatística foi realizada pelo teste do qui-quadrado, teste de Student, ANOVA e a expressão gênica foi analisada com REST®. No experimento 1 bovino as taxas de clivagem foram semelhantes entre os grupos, mas a taxa de blastocisto total foi menor no sistema sequencial e a taxa de blastocisto expandido foi maior no sistema de cultivo único. Além disso, o cultivo de embriões no sistema único resultou em maior expressão de GLUT1 e GLUT5 (P <0,01) e níveis de expressão semelhantes de HSP70.1 e PRDX1 em comparação com os embriões cultivados em meios sequenciais. No experimento 2 bovino tanto a taxa de clivagem quanto a taxa de blastocisto foi semelhante entre os grupos. Para ratos, no experimento 1 98,2% dos embriões coletados foram considerados viáveis. No experimentos 2 o grupo superovulado apresentou ovários com maior peso e maior número de embriões, comparado com o grupo controle. Entretanto, os embriões do grupo superovulado apresentaram menor número de células 72 horas após a administração de hCG. No experimento 3 a taxa de blastocisto foi de 83,17%, com taxa apoptótica de 10.32 ± 8.91%. No experimento 4 a taxa de blastocisto total foi menor em sistema sequencial (13,6%) quando comparado com KSOM (24,0%) e sistema de cultivo único (32,9%)(p<0.05). Concluiu-se que o cultivo de embriões em meio único é mais favorável em comparação com a de meios sequenciais, a superovulação foi bem sucedida no modelo rato e a técnica de TUNEL se mostrou viável na avaliação da qualidade embrionária de embriões de ratas Wistar. / The objective was to evaluate culture systems proposed to use in human embryos: single culture system and sequential culture system, compared with culture used in experimental model bovine and Wistar rats. Specifically we evaluated the cleavage and blastocyst rate, embryo kinetics, quantification of gene expression in bovine blastocysts and evaluated the viability and quality by morphology and Tunel of embryos derived from female rats. For bovine cumulus-oocyte complexes were matured, fertilized in vitro. In experiment 1 zygotes were distributed in culture systems: CR2aa, SOFaa, sequential culture system ECM®/Multiblast® and single culture system Global®. We evaluated cleavage and blastocyst rate and gene expression PRDX, HSP70.1, GLUT1 and GLUT5 in blastocysts. In experiment 2, the embryo development was evaluated on the development of bovine embryos biopsied. The zygotes were distributed in culture systems: CR2aa, sequential culture system G1e/GZ) and ECM®/Multiblast®, single culture system Global®. For all groups, the 8-16 cells embryos were biopsied and returned to culture. We evaluated cleavage and blastocyst rate. For Wistar rats, we used animals of the Center for Biology of Reproduction (CBR/UFJF), with twelve weeks of life and superovulated with 150IU/kg of PMSG and 75 IU/kg hCG intraperitoneally. In experiment 1 the embryo collection proceeded 24 hours after hCG and we evaluated embryo viability In experiment 2 rats were distributed between the control and superovulated and embryos were collected at 48 and 72 h after hCG administration. Weight of ovaries, the total number of embryonic structures and quality of embryos were analyzed. In experiment 3 embryo collection proceeded to 72 h after hCG administration. We evaluated the quality of embryos cultured in vitro from superovulated rat through the blastocyst rate and apoptotic index. Already in experiment 4 embryo collection was performed 48 h after hCG administration. Embryos recovered were randomized according to the groups: sequential culture system ECM®/Multiblast®, single culture system Global®, KSOM. We evaluated the rate of blastocyst. Statistical analysis was performed using the chi-square test, Student's test, ANOVA and gene expression was analyzed with REST®. In experiment 1 bovine cleavage rate was similar between groups, but the total blastocyst rate was lower in the sequential system and expanded blastocyst rate was higher in single culture system. Furthermore, the embryo cultured in single culture resulted in higher expression of GLUT1 and GLUT5 (P <0.01) and similar expression levels of HSP70.1 and PRDX1 compared to embryos cultured in sequential media. In experiment 2 both cleavage andblastocyst rate was similar between bovine groups. For rats in experiment 1, 98.2% collected embryos were were considered viable. In experiments 2 superovulated group showed ovaries with greater weight and greater number of embryos, compared with control group. However, embryos from superovulated group showed smaller number of cells 72 hours after hCG administration. In experiment 3 the blastocyst rate was 83.17%, with 8.91% ± 10:32 apoptotic rate of. In experiment 4 the total blastocyst rate was lower in sequential system (13.6%) compared to KSOM (24.0%) and single culture system (32.9%) (p <0.05). It was concluded that the culture of embryos in single medium is more favorable than sequential media, superovulation was successful in a rat model and the TUNEL technique proved feasible in evaluating the quality of embryos from embryonic Wistar rats.

The thesis deals with Formula Student cooling system. The thesis is divided to three main parts, the first part deal about description of current cooling system and lubricating system. The second part of thesis deals with measuring individual component of system at measuring stand, wind tunnel and measuring at vehicle. The last part of thesis deal about evaluation of measurement and construction changes for improving parameters of the cooling system.

This master’s thesis is a reconnaissance study developed to create a concept of road connection between the road 43 and the highway D52 in the Czech Republic. The possible route of road 43 is planned both in highway and road category in so called Bitysska trail and is connected to highway D1 at Ostrovacice interchange. This thesis suggests highway and road options to connect interchange Ostrovacice and highway D52 at the section between Ledce and Rajhrad. Suggested options were compared and the most convenient draft option was further worked out.

This thesis is focused on the determination of platinum in samples of road dust and soil by atomic absorption spectrometry. The theoretical part describes the basic physical and chemical properties of platinum, platinum function in automotive converters and its behavior in the environment. In the experimental section summarizes the methods used to decompositions samples, concentration of the analyte and the main results of the measurements.

The theme of this thesis is to create a conceptual study of the tunnel on high speed railway. Selection of suitable cross section and tunnelling method with considering of geological conditions of the site. Design of primary support and technological process of construction.

L’excavation d’un tunnel profond dans des terrains poussants pose des difficultés particulières de conception et d’exécution. Ce type de terrain est caractérisé par des fortes convergences en paroi du tunnel de nature différée et souvent anisotrope. Le comportement d’un tunnel excavé en terrain poussant est très influencé par la technique d’excavation utilisée. Le cas d’étude du tunnel routier du Fréjus et de sa galerie de sécurité permet d’illustrer ce phénomène. Il s’agit de deux tunnels parallèles qui montrent une configuration très intéressante étant donné qu’ils traversent des conditions géotechniques similaires et qu’ils sont creusés avec des techniques d’excavation différentes : le tunnel routier a été creusé par méthode conventionnelle à l’explosif tandis que la galerie de sécurité a été creusée avec un tunnelier à bouclier simple. Les mesures d’auscultation réalisées pendant l’excavation des deux tunnels ont été analysées et comparées. Des modélisations numériques pour simuler la réponse des deux tunnels ont été développées avec le logiciel Flac3D. Le comportement du terrain est simulé avec un modèle visco-elasto-plastique et anisotrope. L’anisotropie liée à la schistosité du terrain est introduite dans le modèle par la présence de plans de faiblesse d’orientation donnée (ubiquitous joint model) insérés dans une matrice rocheuse caractérisée par un comportement visco-elasto-plastique isotrope. Une rétro-analyse a été réalisée sur les mesures de convergence obtenues lors du creusement du tunnel routier du Fréjus. Le comportement du terrain identifié dans le tunnel routier est ensuite extrapolé pour prédire la réponse de la galerie de sécurité. L’objectif est de reproduire l’état des contraintes observé dans les voussoirs de la galerie de sécurité et d’extrapoler les sollicitations à long terme. L’influence que la technique d’excavation, en particulier sur le comportement différé du terrain a été prise en compte dans les simulations numériques. On a mis en évidence que les déformations différées du terrain sont réduites lorsque l’excavation est réalisée au tunnelier.Par ailleurs, une synthèse critique de la méthode convergence-confinement et de ses variantes a été réalisée. Une discussion a été menée sur l’applicabilité des méthodes convergence-confinent quand elles sont utilisées pour le dimensionnement des tunnels circulaires excavés en section pleine avec l’installation d’un soutènement raide près du front d’excavation comme c’est le cas lors d’une excavation au tunnelier. Dans ce contexte, un ensemble de formules empiriques sont proposées. Elles permettent d’obtenir avec une bonne précision l’état d’équilibre entre le terrain et le soutènement et peuvent être utilisées dans la phase de pré-dimensionnement des ouvrages / During the excavation of deep tunnels, squeezing ground conditions are often encountered. The squeezing behavior of the ground is characterized by large time-dependent and usually anisotropic convergences that take place at the tunnel wall. The technique of excavation has a strong influence on the tunnel response when it is excavated under squeezing conditions. This phenomenon is illustrated throughout the case study of the Fréjus road tunnel excavated with conventional drill and blast methods and of its safety gallery excavated with a single shield tunneling boring machine. They exhibit a very interesting configuration of two tunnels excavated in parallel under the same geotechnical conditions but with different excavation techniques. Monitored geotechnical data from both tunnels are analyzed and compared. Numerical simulations of both tunnels have been carried out with Flac3D. An anisotropic creep model which includes weakness planes of given orientation embedded in a visco-elasto-plastic matrix has been used for describing the behavior of the ground. A back-analysis of convergence measurements of the Fréjus road tunnel has been carried out. The behavior of the ground identified from the Fréjus road tunnel is extrapolated to predict the response of the Fréjus safety gallery in terms of the stress state in the lining. The influence of the technique of excavation on the time-dependent parameters of the ground is taken into account in the computations and its effects are discussed. It is shown that the long term ground deformation are significantly reduced with TBM excavation as compared to traditional blast and drill method.Furthermore, the convergence-confinement methods are reviewed and their applicability is discussed when they are applied to full face circular tunnels excavated in rock masses with a stiff support system near the face. In this context, a set of empirical formula are proposed which allows to accurately predict the equilibrium state between the ground and the lining in circular tunnels excavated in full section. These formula are useful in the preliminary phase of tunnel design

Cleft lip is one of the most common human birth deffects. Its etiopathogenesis is multifactorial and many aspects of its occurrence remain unknown in the fields of both genetics and teratology. One of the set of known negative external factors causing cleft lip is chemical hydrocortisone. Its effect on cell proliferation is highly heterogeneous and depends on attributes of a specific cell population. In this work we studied the cleft beak origin after the hydrocortisone treatment on the basis of Chick Embryotoxicity Screening Test (CHEST). Our main aim was to detect cell cycle changes in the chick frontonasal process after hydrocortisone injection via flow cytometry analysis. Hydrocortisone caused S phase arrest within a minor subpopulation of highly granular cells with specific cell cycle. This sensitive subpopulation was localized in the areas of previously defined proliferative centers within the frontonasal process using immunohistochemistry of frozen sections. Quantitative analysis of cells in these areas revealed significant decrease of M phase portion in the hydrocortisone treated samples in comparison with the control samples. The TUNEL staining of histological sections was used to determine the apoptotic rate in the frontonasal process. The comparison between the control and the...