Meiosis is a specialized cell division that produces haploid gametes from a diploid progenitor cell. It consists of one round of DNA replication followed by two consecutive rounds of chromosome segregation. Homologous chromosomes segregate in meiosis I and sister chromatids segregate in meiosis II. Failure to correctly regulate meiosis can result in aneuploidy, where daughter cells inherit an incorrect number of chromosomes. Aneuploidy is usually poorly tolerated in eukaryotes, and is associated with infertility, miscarriages and birth defects. At the meiosis I to meiosis II transition, DNA replication does not occur between chromosome segregation steps despite the need for Spindle Pole Bodies (SPBs) to be re-licensed in order to build meiosis II spindles. The mechanisms that make this distinction are not yet known. In budding yeast, the protein phosphatase Cdc14 is essential for the progression of cells into meiosis II. Cdc14 is sequestered for the majority of the cell cycle in the nucleolus by the inhibitor Cfi1/Net, and is only released in anaphase. We have observed Cdc14 localizing to and interacting with SPB components when nucleolar sequestration is inhibited. Through fluorescence microscopy and EM analysis, we have determined that Cdc14 is required for the re-duplication of SPBs after meiosis I. Our data implies a role for Cdc14 in the phospho-regulation of SPB half-bridge component Sfi1. Cdc14 is therefore essential for the relicensing of SPB duplication, a crucial step necessary to ensure accurate chromosome segregation in meiosis.

Le centrosome est le centre organisateur des microtubules dans les cellules animales, il nucléé les microtubules interphasiques ainsi que le fuseau mitotique. Les centrosomes sont produits par duplication, mécanisme rigoureusement régulé au cours du cycle cellulaire. En effet, un centrosome comporte deux centrioles qui se dupliquent une fois par cycle cellulaire. Des erreurs de duplication conduisant à plus de deux centrosomes induisent la formation de fuseaux multipolaires et provoquent des défauts de ségrégation des chromosomes. Chez la levure Schizosaccharomyces pombe, un organisme modèle pour l’étude de la division cellulaire, les homologues des centrosomes sont les SPBs (pour Spindle Pole Body). Une structure annexe spécifique liée aux SPBs est appelée demi-pont (quand les SPBs ne sont pas dupliqués) puis pont (quand elle relie les deux SPBs dupliqués). Les deux principaux composants du pont chez la levure S. pombe sont Cdc31 et Sfi1. Sfi1 est une protéine linéaire formée de répétitions en hélice α formant des sites de liaison pour la Centrine/Cdc31. Sfi1 s’assemble en réseau de molécules parallèles interagissant avec le SPB via leur domaine N-terminal. Lors de la première partie de ma thèse, j’ai démontré que Sfi1 est requis pour la duplication et la séparation des deux SPBs. Dans la première partie de ma thèse, je me suis intéressée aux fonctions de Sfi1 chez la levure. Cette étude a permis de démontrer que Sfi1 est un composant du demi-pont et qu’il est essentiel pour la duplication des SPBs et l’assemblage d’un fuseau bipolaire. De plus, nous avons déterminé que le pont est dupliqué en fin de mitose. Enfin, nous avons aussi montré que la déstabilisation du pont menant à sa rupture en mitose, dépend de la phosphorylation de Cdc31 par la kinase mitotique Cdk1. Lors de la seconde partie de ma thèse, je me suis intéressée au complexe Sfi1/Centrine dans les cellules humaines. J’ai confirmé que Sfi1 est localisée aux centrioles. De plus, j’ai montré que la déplétion de Sfi1 dans les cellules RPE1, conduit à une perte de localisation de la Centrine, suggérant soit un défaut de recrutement, soit une déstabilisation. De plus, en absence de Sfi1, les cellules RPE1 ne sont plus capables de former de cil primaire. Ce résultat suggère que Sfi1 et la Centrine sont requis pour la ciliogénèse. Enfin, j’ai aussi démontré que la déplétion deSfi1 induit un arrêt de cycle cellulaire dans les cellules non tumorales RPE1. Dans les cellules cancéreuses, HeLa, le cycle n’est pas arrêté mais j’ai pu observer une prolongation du temps de mitose. En conclusion mes travaux montrent que bien que la fonction de Sfi1/Centrin ne soit pas conservée, le complexe reste essentiel pour l’intégrité structurale et fonctionnelle du centrosome. / The centrosome is the main microtubule organizing center. It nucleates and organizes interphase microtubule and contributes to the assembly of the bipolar mitotic spindle. To do so, the centrosome, present in one copy at the beginning of the cell cycle, duplicates to produce a second copy. The duplication process is tightly controlled and regulated since centrosome over-duplication can lead to multipolar mitotic spindles and promote genome instability and tumorigenesis. The duplication of the yeast centrosome, the SPB (Spindle pole body), begins with the duplication of the half bridge. This appendage is composed of Sfi1/Cdc31 complex organized in a parallel array attached to the core SPB. SPB duplication relies on the assembly of a second array of Sfi1/Cdc31, anti-parallel to the first one, creating thereby an assembly site for the new SPB. Therefore Sfi1 is essential for SPB duplication and our work defined the timing of half-bridge duplication and some of the regulatory mechanisms that favor bridge splitting to release duplicated centrosomes and allow spindle assembly at mitotic onset. Sfi1 and Cdc31/Centrins are conserved in human cells where the centrosome is composed of two centrioles surrounded by the pericentriolar material. Centrins are concentrated in the distal end of centrioles. Sfi1 has also been localized to centrioles, but its function remained unknown. Thus, we started investigating Sfi1 function in human cells. We found that Sfi1 depletion leads to a decrease in Centrin recruitment to the centrioles. It also leads to a cell cycle arrest in G1 in RPE1 cells, an event previously observed in presence of defects in centriole biogenesis. In HeLa cells where the cell cycle is not affected, Sfi1 depletion leads to a mitotic delay. Moreover, Sfi1 depletion leads to cilium assembly. To conclude, these results altogether point towards a role of human Sfi1 in centriole biogenesis.

This dissertation focuses on three research projects: 1) construction of simultaneous prediction intervals/bounds for at least k out of m future observations; 2) semi-parametric degradation model for accelerated destructive degradation test (ADDT) data; and 3) spatial variable selection and application to Lyme disease data in Virginia. Followed by the general introduction in Chapter 1, the rest of the dissertation consists of three main chapters. Chapter 2 presents the construction of two-sided simultaneous prediction intervals (SPIs) or one-sided simultaneous prediction bounds (SPBs) to contain at least k out of m future observations, based on complete or right censored data from (log)-location-scale family of distributions. SPI/SPB calculated by the proposed procedure has exact coverage probability for complete and Type II censored data. In Type I censoring case, it has asymptotically correct coverage probability and reasonably good results for small samples. The proposed procedures can be extended to multiply-censored data or randomly censored data. Chapter 3 focuses on the analysis of ADDT data. We use a general degradation path model with correlated covariance structure to describe ADDT data. Monotone B-splines are used to modeling the underlying degradation process. A likelihood based iterative procedure for parameter estimation is developed. The confidence intervals of parameters are calculated using the nonparametric bootstrap procedure. Both simulated data and real datasets are used to compare the semi-parametric model with the existing parametric models. Chapter 4 studies the Lyme disease emergence in Virginia. The objective is to find important environmental and demographical covariates that are associated with Lyme disease emergence. To address the high-dimentional integral problem in the loglikelihood function, we consider the penalized quasi loglikelihood and the approximated loglikelihood based on Laplace approximation. We impose the adaptive elastic net penalty to obtain sparse estimation of parameters and thus to achieve variable selection of important variables. The proposed methods are investigated in simulation studies. We also apply the proposed methods to Lyme disease data in Virginia. Finally, Chapter 5 contains general conclusions and discussions for future work. / Ph. D.

The goal of this master's thesis was to develop a method for gas chromatography using the Controlled Evaporation Mixer (CEM) for gas mixture sampling. Gas mixture contained chemical substances formed during manufacturing of acrylic acid by two-step oxidation of propylene. The analyzed mixture composed of permanent gases, light hydrocarbons and compounds containing oxygen. During the method development several capillary columns with polar and nonpolar stationary phase were used. After selection of the most suitable column (SPB-1 Sulfur, Supelco, USA) the method was further optimized for the maximal possible separation efficiency. However, it was impossible to separate propane and propene on SPB-1 Sulfur column. These gases were possible to determine on TCD detector and separate on respective columns prior to the detector. For that purpose, the optimization of multiple-port valves switching inside the gas chromatograph was performed. The next step was calibration of gas chromatograph for a purpose of quantitative analysis of selected substances. The calibration solution containing solvent and an analyte was evaporated and diluted with nitrogen by CEM to obtain desired analyte concentration. Formaldehyde and propane had a very low response on detectors, acetaldehyde was undetectable in desired...

Meiosis is the process by which haploid gametes are produced from a diploid cell. It is a specialised form of cell division which involves one round of DNA replication followed by two rounds of chromosome segregation. Errors in the segregation process can give rise to aneuploidy, which can result in miscarriages and birth defects. In the first meiotic division homologous chromosomes are segregated, and sister chromatids are segregated in the second division. This is coordinated with two rounds of spindle microtubule assembly and disassembly. How these two processes are coordinated is unknown. In my PhD, I study the role of the protein phosphatase 2A (PP2A) regulatory subunit, Cdc55, in budding yeast meiosis. PP2A is a conserved heterotrimeric enzyme that has important roles in mitosis and meiosis. These roles are dictated by binding to either of its two regulatory subunits, Rts1 and Cdc55, in budding yeast . I show that Cdc55 is required for the proper assembly of a meiotic spindle in meiosis I, through the maintenance of the Cdc14 phosphatase in the nucleolus early in meiosis. In addition, Cdc55 is also required to limit the formation of PP2A complexes with the Rts1 regulatory subunit, and this is essential for the timely dissolution of sister chromatid cohesion. Thus, Cdc55 couples spindle assembly with chromosome segregation through its interactions with Cdc14 and PP2ARts1. Finally, I show some preliminary studies looking at the possible downstream effectors of Cdc14 that are important in this mechanism.

Southern pine beetle (SPB), Dendroctonus frontalis (Zimmerman), is one of the most destructive insect pests of pine trees in southern United States, Mexico and Central America. There is relatively little information on the effect of habitat fragmentation on the connectivity and the spatial population genetics of SPB. This study therefore, adds to previously generated information by assessing how habitat fragmentation affects the spatial population genetic structure of SPB. It also introduces a new approach to the study of bark beetle population dynamics by assessing how landscape variables shape their effective dispersal. To address this issue, a suite of eight highly polymorphic DNA microsatellite markers were used to measure SPB movement over a representative range of SPB habitat and non-habitat (matrix). At the broadest scale, highly significant genetic differentiation suggests that the sampled populations are not panmictic. Loci with higher variability yielded higher resolution for both the infinite allele model based measure of differentiation (FST) and the stepwise mutation based measure (RST) estimates. It is apparent that allelic frequency differences, allelic size ranges and repeat motif played a role in the observed patterns of pair-wise differentiations between the sampled localities. It is supposed that gene flow, wide-range dispersal and recent divergent time could have contributed to the lower level of genetic structure observed in the pair wise estimates. The sampled populations did not show any differentiation attributable to the host species from which they were collected. Mantel test of genetic distance and Euclidean geographic distance revealed no correlation. Mantel tests of the correlation between genetic distance and cost weighted Euclidean distances also suggest that dispersal of SPB across geographic barriers is not significantly reduced. Thus, landscape features and host preference do not appear to have had an impact on population genetic structure of SPB. Since movements of these beetles were not significantly hindered by environmental factors like major rivers, roads, elevation and host type, it is advisable for regional pest management offices to put an effort and coordinate their prevention and management plans in a broader scale to alleviate the problem associated with this native insect pest. Disequilibrium in the observed homogenized pattern of the beetle in this study suggests human contribution in the dispersal of SPB. Therefore, stringent control is deemed necessary in transportation of logs. This could improve our pest management system hence its positive implication in timber industry is obvious.

Noise is a growing health concern as urban residents increases rapidly and more reports of noise causing sleep disturbances and increasing the risk of cardiovascular health problems are published. Noise has a negative influence on life quality. This life quality deficiency also shows in housing and office pricing in noisy environments. Housing and office prices are often higher in quiet areas than in noisy areas. Therefore noise is both a health issue and has big economic consequences. The biggest contributor to the road traffic noise is the tire/road noise at speeds above 50km/h. Therefore this is an important aspect to monitor and the problem has to be alleviated. In this thesis the client Ramböll has gotten many contracts concerning the state of the roads from governmental institutes. As a part of the evaluation of the roads they want to implement the noise emission as a factor. This gives the government institutions another parameter that helps in the decision of which part of the road network to refurbish first. In the effort to decide what is best way for Ramböll to measure noise a new setup for measuring noise has been developed. The most used ways of measuring tire/road-noise is the close proximity (CPX) -method and the statistical pass-by (SPB) -method. These methods both give accurate results but they have drawbacks. The SPB measurements are time consuming and only give noise levels for a small patch of a road. CPX measurement on the other hand require costly and time consuming development of a measurement trailer. Certifying the trailer and maintenance work of it is expensive. So this report shows a first step in how to build a measurement setup and what aspects were taken into consideration when it was designed. A close proximity measurement setup in the form of a tube with a microphone placed inside it was built and installed underneath the measurement vehicle. The  measurement setup designed and built was named the Tube-CPX measurement setup in this report. The measurements performed with this Tube-CPX measurement setup show promising results. Similarities between CPX measurement setup and the Tube-CPX setup have been found both in the frequency spectrum as well as in the relation with pass-by measurement noise levels. The repeatability of the Tube-CPX measurements is even better than the compared CPX measurements. Although results are promising more work is required before the Tube-CPX setup can be seen in operation at Ramböll. In particular the question which source causes which sound pressure is required to understand the measured levels. And also more controlled pass-by measurements have to be performed to determine the relation of the absolute values measured at the tire/road impact spot to the values that are most interesting, namely the sound pressure levels that the human at the side of the road depicts.

In this study, two symbiotic fungi of Southern Pine Beetle (SPB), Entomocorticium peryii and Entomocorticium sp.A were evaluated in terms of polyphenol oxidase (PPO) production. The effect of different inhibitors, inducers and assay parameters such as temperature and pH on enzyme activity were investigated and maximum PPO activity was observed at 30&deg / C, pH 8.0 and when tannic acid was used as an inducer. Copper-chelator salicyl hydroxamic acid (SHAM) and pcoumaric acid, both indicated as inhibitors of tyrosinase and catechol oxidase significantly reduced the activity. For biochemical characterization studies, the enzyme was concentrated by ultrafiltration. To determine type of the enzyme, activity staining after Native-PAGE was carried out. Type of polyphenol oxidase produced by E. peryii and E. sp.A was determined as catechol oxidase by activity staining. However higher activity was observed on hydroquinone (p-diphenol) rather than catechol (o-diphenol). The enzyme obeys Michealis-Menten kinetics with Km and Vmaxvalues being 10.72 mM hydroquinone and 59.44 U/ml for E. peryii and 8.55 mM hydroquinone and 73.72 U/ml for E. sp.A respectively..

L’école est fréquemment identifiée comme un milieu important dans la pratique de saines habitudes de vie des jeunes. Cette recherche s’intéresse à l’influence de l’environnement bâti proximal de l’école sur la pratique de l’activité physique d’élèves du Québec. L’échantillon utilisé inclut 11 207 élèves de 142 écoles primaires publiques québécoises. L’environnement bâti a été caractérisé à l’aide d’indicateurs géographiques. L’association entre l’activité physique des élèves et les caractéristiques environnementales a été estimée à l’aide de modèles de régression logistique multiniveaux. Les analyses démontrent l’existence de différences entre les écoles dans le niveau d’activité physique des élèves. La densité de végétation mesurée dans le voisinage est associée positivement à l’activité physique. Les inégalités environnementales à l’échelle des écoles semblent induire des différences dans les habitudes de vie des élèves. Les caractéristiques du design urbain étant modifiables, celles-ci constituent de possibles pistes d’interventions visant à accroître l’activité physique chez les jeunes.

This thesis focuses on redundancy in data networks and on technologies which are used to achieve high availability in network infrastructure. This thesis is based on industry standards published by well-known standardization authorities. One proprietary technology which can effectively replace these standards and simplify the whole network is also presented in more detail. The theoretical part individually describes all such technologies and standards. The practical part deals with an appropriate combination of these technologies and their application in a specific product environment. This work is meant to describe the whole concept of high availability in network infrastructure and should be instrumental in understanding how these technologies are deployed in practice.