Klimawandelmodelle sagen für Deutschland sowohl höhere Jahresdurchschnittstemperaturen als auch eine Abnahme von Niederschlägen in den Sommermonaten voraus. Mögliche Konsequenzen für Bäume sind eine verlängerte Vegetationsperiode, ein erhöhtes Spätfrostrisiko und mehr Trockenstress während des Sommers. Diese veränderten Umweltbedingungen könnten zu Veränderungen der Konkurrenzverhältnisse zwischen Baumarten führen. Die Rotbuche (Fagus sylvatica L.) ist eine der wichtigsten Laubbaumarten Mitteleuropas. Daher ist das genetische Anpassungspotential dieser Baumart an den Klimawandel von großem Interesse. In dieser Studie wurden sowohl die neutrale als auch die adaptive genetische Variation der Buche untersucht. Dafür wurde ein Translokationsexperiment mit Nachkommen von Buchenpopulationen, die unter verschiedenen Umweltbedingungen in Norddeutschland wachsen, etabliert. Wiederholte Aufnahmen wichtiger phänotypischer Merkmale (Höhe, Austrieb, Trockenstresssensitivität, Sterblichkeit) zeigten signifikante Unterschiede zwischen den Populationen. Interessanterweise zeigten Populationen mit einer größeren geographischen Distanz teilweise ähnlichere Phänotypen als benachbarte Populationen. Die neutrale genetische Variation der untersuchten Sämlingspopulationen wurde anhand neun verschiedener Mikrosatellitenmarker analysiert. Zwischen den analysierten Buchenpopulationen wurde nur eine geringe genetische Differenzierung ermittelt. Die genetische Diversität war hoch und statistisch nicht signifikant unterschiedlich von den Altbeständen, aus denen sie stammten. Die hohe genetische Diversität ist eine gute Basis für Adaption, allerdings könnte sie wahrscheinlich nur eine kurzfristige Anpassung an den Klimawandel ermöglichen. Daher ist es wichtig, Einblicke in die genetische Basis von klimawandelrelevanten Merkmalen zu gewinnen. Deshalb wurden in dieser Studie Kandidatengene für das Austriebsverhalten untersucht. Bei der Analyse von Fragmenten von zehn verschiedenen Kandidatengenen wurden 20 Indels und 116 SNPs identifiziert. Insgesamt wurden 46 SNPs erfolgreich zur Genotypisierung von über 1.400 Individuen, die aufgrund ihres Austriebsverhaltens ausgewählt wurden, verwendet. Assoziationsanalysen wurden durchgeführt, um potentiell adaptive SNP-Marker zu identifizieren. Diese ergaben unter einem „generalisierten linearen Modell“ 23 signifikant mit dem Austrieb assoziierte SNPs. Ein zusätzlich verwendetes „gemischtes lineares Modell“ ergab nahezu gleiche Ergebnisse. Die phänotypische Variation, die durch signifikant mit dem Austrieb assoziierte SNPs erklärt wird, war niedrig (R2 < 2,2), aber in Übereinstimmung mit anderen Studien mit Waldbaumarten. Zusätzlich zu den Assoziationsanalysen wurden auch FST-Outlier-Analysen durchgeführt. Diese ergaben sieben verschiedene SNPs, die potentiell unter ausgleichender oder gerichteter Selektion stehen. Insgesamt wurden vier potentiell adaptive SNPs gleichzeitig durch Assoziations- und Outlier-Analysen identifiziert. Diese könnten die höchste Wahrscheinlichkeit aufweisen, an der Ausprägung des Austriebsverhaltens beteiligt zu sein. Allerdings sind viele potentiell adaptive SNPs, die in dieser Studie identifiziert wurden, nicht-kodierend oder synonym und somit nicht die kausativen SNPs, sondern eher gelinkt mit ihnen. Allerdings wurde in dieser Studie ein geringes Kopplungsungleichgewicht (linkage disequilibrium) gefunden. Somit könnten die kausativen SNPs in naher Umgebung liegen. Die in dieser Studie identifizierten potentiell adaptiven SNPs sollten in weiteren Studien mit zusätzlichen Populationen bestätigt werden.

The overall aim of this thesis was to investigate the genetic control of beef quality in a commercial population of Aberdeen Angus-sired cattle with a view to trait improvement. The population studied included 500 Angus-cross animals, all with purebred Aberdeen Angus sires, from a selection of farms throughout Scotland. A number of carcass-related weight traits and taste panel assessed sensory traits were measured on these animals. A population of 265 Charolais cross cattle (all with purebred sires) was then used to explore the extrapolation of results across breeds. The first aim of this thesis was to investigate heritabilities for important carcass and meat quality traits and to assess the quality of a number of taste-panel derived meat quality traits by calculating three consistency statistics. Consistency statistics (parameter range 0 to 1) for the taste panel traits were moderately high, particularly for panel member consistency and reproducibility, with values ranging from 0.48 to 0.81 and 0.43 to 0.73, respectively. Estimated heritabilities were low for most of the sensory taste-panel-evaluated traits, where the maximum value was 0.16 for overall liking, but were higher for carcass traits where carcass weight heritability was 0.7. To perform these analyses it was first necessary to confirm paternity using a number of genetic markers. Therefore, a comparison of the power of both microsatellite and SNP markers for paternity exclusion was carried out to determine the more effective method. Results indicated that approximately three times as many SNP markers than microsatellite markers were required for parentage exclusion, and a panel of 15 microsatellite markers was used to assign paternity before subsequent data analysis was carried out. The remaining aims of this thesis centred on exploring genetic markers for carcass and meat quality. Firstly, the Angus animals were genotyped for the del11 myostatin mutation which was found to be segregating at a relatively low frequency (0.04) and was shown to be associated with a 17.4 kg increase in carcass weight (P < 0.05) in the heterozygous animals when compared to the homozygous wild-type animals. By analysing the haplotype associated with the mutant allele, it was determined that there have been at least two separate introductions of the mutant allele into the Aberdeen Angus breed. A number of SNPs were also tested for their effects on the carcass and meat quality traits in the Angus animals. The SNPs fell into two groups: eight that have been incorporated into commercially available tests and a further 28 from alternative candidate genes that have effects in different breeds and species. In total, 17 SNPs significantly affected at least one of the traits measured. Of these significant associations, a number have been seen previously, such as the association between calpain and tenderness (P = 0.01) and growth hormone and eye muscle area (P = 0.05), and some of which were novel, such as the association between growth hormone receptor and steak odour (P = 0.02) and corticotrophin releasing hormone and gristle distance from fat (P = 0.004). A further six SNPs, identified by resequencing of the malic enzyme 1 (ME1) and small heterodimer partner (SHP) genes, were tested for their effects on the traits measured in this thesis. Five of the SNPs, including one which caused a non-synonymous amino acid change, had a significant effect on at least one of the traits tested including fat class (P = 0.002), eye muscle area (P = 0.01), sirloin weight before maturation (P = 0.03), sirloin steak tail length (P = 0.004) and juiciness (P = 0.004) where the effect sizes were 1.79 units, 565 mm2, 0.36 kg, 17.12 mm and 0.23 taste panel units, respectively. To assess the effect of the genotyped SNPs on intramuscular fat (IMF), a simple method of visible IMF quantification in the sirloin steak was developed using digital photographs and an image analysis program. Results showed that two SNPs in the calpain gene, known to be linked with an increase in meat tenderness, were associated with an increase in visible IMF% and the del11 mutation was associated with a reduction in visible IMF%. The heritabilities, SNP association validations and novel SNP-trait associations identified in this thesis provide tools for use in breeding programs, possibly via marker assisted selection to improve meat quality traits. However, the results seem breed-specific, as most of the significant effects were not replicated in the Charolais population.

Le tocophérol, le phytostérol, le pourcentage de protéines des graines, l'huile et les teneurs en acides gras ont été mesurés dans une population de lignées recombinantes (RILS) de tournesol, cultivées sous conditions de sécheresse, irrigation et semis tardif. Une analyse génétique de QTL a été réalisée à partir de ces mesures, en utilisant une carte génétique basée sur des marques SSR et avec des gènes candidats (1) impliqués dans la voie métabolique de tocophérol et phytostérol, (2) des gènes codant des antioxydants enzymatiques, (3) des gènes liés à la sécheresse et (4) des gènes homologues à SEC14 chez Arabidopsis. Trois gènes candidats importants (VTE4, VTE2 et HPPD), qui codent pour des enzymes impliquées dans la biosynthèse du tocophérol, ont été cartographiés sur les groupes de liaison LG8 et LG14. Quatre SNPs sont identifiés pour PAT2, le gène homologue chez Arabidopsis SEC14, entre les deux parents (PAC2 et RHA266) et un SNP, identifié par alignement de séquences est converti en marqueur CAPS pour permettre l'analyse génotypique des RIL. Les gènes homologues à SFH3, HPPD, CAT et CYP51G1 ont été cartographiés grâce à la mise au point de marqueurs dominants, tandis que des marqueurs co-dominants ont permis la cartographie des gènes homologues à SEC14-1, VTE4, DROU1, POD, SEC14-2 et AQUA. Les gènes POD, CAT et GST, codant pour des antioxydants enzymatiques, ont également été cartographiés sur les groupes de liaison 17, 8 et 1, respectivement. Le QTL majeur pour la teneur en tocophérol a été identifié sur le groupe de liaison 8, qui explique 59,5% de la variation phénotypique (6.TTC.8). Il colocalsie également avec le QTL identifié pour la teneur en phytostérol (7.TPC.8). Sous condition de semis tardif, un QTL spécifique de la teneur en acide palmitique a été identifié sur le groupe de liaison 6 (PAC-LS.6). Il est situé entre les marqueurs ORS1233 et SSL66_1. Les QTLs pour le pourcentage d'huile de graines et la teneur en acide stéarique colocalisent sur les groupes de liaison 10 (PSO-PI.10 et SAC-WI.10) et 15 (PSO-PI.15 et SAC-LS.15). Sept QTLs associés à teneur en acides palmitique, stéarique, oléique et linoléique sont identifiés sur le groupe de liaison 14. Ils sont liés à l’homologue du gène HPPD. Par ailleurs, les caractères agronomiques tels que les jours du semis à la floraison, la hauteur des plantes, le rendement et la morphologie foliaire ont été étudiés. Des analyses association génétique ont permis d’identifier des QTLs intérêts sur les groupes de liaison 2, 10 et 13 pour les caractères étudiés, d’autres QTLs identifies sur les groupes de liaison 9 et 12 mettent en avant l'importance de ces régions génomiques pour les caractères de morphologie foliaire. Nous avons finalement identifié des marqueurs AFLP et quelques gènes candidats liés aux caractères impliqués dans la qualité des graines sous conditions irriguée et stress hydrique chez une population de mutants (M8). Deux lignées mutantes, M8-826-2-1 et M8-39-2-1, produisent un niveau significativement élevé d'acide oléique peuvent être utilisées dans les programmes de sélection en raison de la haute stabilité à l'oxydation et des propriétés cardiovasculaire apportés par l’acide oléique qu’elles produisent. L'augmentation du niveau de tocophérol dans les lignées mutantes, M8-862-1N1 et M8-641-2-1, est justifiée par le polymorphisme observé pour le gène, MCT, impliqué dans la voie métabolique du tocophérol. Le marqueur le plus important pour le contenu en tocophérol total est E33M50_16 qui explique 33,9% de la variation phénotypique. Un des gènes candidats les plus importants concernant la biosynthèse des acides gras, FAD2 (FAD2-1), est lié à la teneur en acides oléique et linoléique. Il explique plus de 52% de la variation phénotypique. / The genetic control of tocopherol, phytosterol, percentage of seed protein, oil and fatty acids content in a population of recombinant inbred lines (RILs) of sunflower under various conditions are studied through QTL analysis using genetic-linkage map based on SSR markers and introducing some important tocopherol and phytosterol pathway-related genes, enzymatic antioxidant-related genes, droughtresponsive family genes and Arabidopsis SEC14 homologue genes. Three important candidate genes (HPPD, VTE2 and VTE4), which encode enzymes involved in tocopherol biosynthesis, are mapped to linkage group 8(LG8) and LG14. One of the most important candidate genes coding for sterol methyltransferase II (SMT2) enzyme is anchored to LG17 by CAPS marker. Four SNPs are identified for PAT2, Arabidopsis Sec14 homologue gene, between two parents (PAC2 and RHA266). PAT2 is assigned to LG2 by CAPS marker. Squalene epoxidase (SQE1) is also assigned to LG15 by InDel marker. Through other candidate genes, POD, CAT and GST encoding enzymatic antioxidants are assigned to LG17, LG8 and LG1, respectively. The major QTL for total tocopherol content on linkage group 8 accounted for 59.5% of the phenotypic variation (6.TTC.8), which is overlapped with the QTL of total phytosterol content (7.TPC.8). Under late-sowing condition, a specific QTL of palmitic acid content on linkage group 6 (PAC-LS.6) is located between ORS1233 and SSL66_1 markers. Common chromosomic regions are observed for percentage of seed oil and stearic acid content on linkage group 10 (PSO-PI.10 and SACWI. 10) and 15 (PSO-PI.15 and SAC-LS.15). Overlapping occurs for QTLs of oleic and linoleic acids content on linkage groups 10, 11 and 16. Seven QTLs associated with palmitic, stearic, oleic and linoleic acids content are identified on linkage group 14. These common QTLs are linked to HPPD homologue, HuCL04260C001. QTLs controlling various traits such as days from sowing to flowering, plant height, yield and leaf-related traits are also identified under well-, partial-irrigated and late-sowing conditions in a population of recombinant inbred lines (RILs). The results do emphasis the importance of the role of linkage group 2, 10 and 13 for studied traits. Genomic regions on the linkage group 9 and 12 are important for QTLs of leaf-related traits in sunflower. We finally identified AFLP markers and some candidate genes linked to seed-quality traits under well-irrigated and water-stressed conditions in gammainduced mutants of sunflower. Two mutant lines, M8-826-2-1 and M8-39-2-1, with significant increased level of oleic acid can be used in breeding programs because of their high oxidative stability and hearthealthy properties. The significant increased level of tocopherol in mutant lines, M8-862-1N1 and M8- 641-2-1, is justified by observed polymorphism for tocopherol pathway-related gene; MCT. The most important marker for total tocopherol content is E33M50_16 which explains 33.9% of phenotypic variance. One of the most important candidate genes involving fatty acid biosynthesis, FAD2 (FAD2-1), is linked to oleic and linoleic acids content and explained more than 52% of phenotypic variance.

La maladie d’Alzheimer est une maladie neurodégénérative, elle représente 70% des formes de démences et affecte près de 860 000 personnes en France. Cette maladie est caractérisée par deux lésions neuropathologiques : les Dégénérescences neurofibrillaires et les Plaques séniles. Ces dernières sont principalement constituées de peptides amyloïdes (A&#946;) résultant du clivage d’une protéine membranaire appelée Précurseur du peptide amyloïde (APP). L’étude des formes familiales monogéniques a montré que des mutations des gènes de l’APP et des Présénilines 1 et 2 conduisaient systématiquement à une augmentation de la production d’A&#946;. Cette observation a permis l’élaboration de la cascade amyloïde plaçant le métabolisme de l’APP au centre du processus physiopathologique. Même si aujourd’hui ce métabolisme commence à être relativement bien connu, plusieurs zones d’ombres subsistent encore. Dans l’optique de caractériser de nouveaux acteurs intervenant dans ce métabolisme, nous avons émis une hypothèse qui repose sur deux constatations : (i) les protéines impliquées dans l’étiologie de la maladie sont différentiellement exprimées entre les cerveaux des patients et ceux des témoins (ii) dans le cerveau, de nombreuses métalloprotéases participent aux même mécanismes que l’APP (adhésion cellulaire, neuroinflammation, plasticité neuronale...), certaines sont aussi directement actrices du métabolisme de l’APP en tant qu’&#945;-sécrétase (ADAM9, ADAM10 et ADAM17) ou en dégradant l’A&#946; (NEP, IDE, MMP2, MMP3 et MMP9). Nous avons donc supposé que les métalloprotéases présentant une différence d’expression entre le tissu cérébral des malades et celui des témoins soient des candidates intéressantes pour moduler le métabolisme et le trafic de l’APP. Une première analyse transcriptomique par biopuce, à partir d’ARN totaux issus des cerveaux de 12 malades et de 12 témoins, nous a permis d’identifier quatre métalloprotéases présentant une différence d’expression significative (p<10-5) : ADAMTS16, ADAM17, ADAM30 et ADAM33. Nous avons cherché à confirmer ce résultat par une autre technologie sur un plus grand nombre d’échantillons (malades n=52 et témoins n=42). Seules ADAM30 et ADAM33 ont pu être validées. Nous avons également pu observer que l’expression d’ADAM30 dans le tissu cérébral des malades est inversement proportionnelle à la quantité d’A&#946;42 déposée dans la parenchyme (A&#946;42 la forme d’A&#946; la plus neurotoxique). De plus, au niveau cérébral, l’expression d’ADAM30 est restreinte aux neurones, cellules sièges du métabolisme de l’APP. Nous avons donc sélectionné ADAM30 comme intervenante potentielle dans le métabolisme de l’APP. Pour tester notre hypothèse, nous avons sous- et sur-exprimé ADAM30 dans deux modèles cellulaires différents. Nous avons mis en évidence que la sur-expression d’ADAM30 entraîne une diminution de l’ensemble des produits du métabolisme de l’APP. En mutant le site catalytique de cette protéase, nous avons remarqué que cette action sur le métabolisme de l’APP est dépendante de cette activité catalytique. De manière cohérente, une sous-expression d’ADAM30 entraîne une augmentation de l’ensemble des produits du métabolisme de l’APP. En utilisant les inhibiteurs alcalisant, nous avons démontré que l’effet d’ADAM30 sur le métabolisme de l’APP met en jeu la dégradation par le lysosome. Des expériences d’immunofluorescence ont attesté qu’ADAM30 est localisée dans le réticulum endoplasmique et l’appareil de Golgi et qu’elle co-localise fortement avec l’APP dans ces organites. Au vu des résultats obtenus durant ces quatre années, nous pensons qu’ADAM30 pourrait être une protéine clé de la régulation de l’APP en inhibant son acheminement jusqu’à la membrane plasmique et en favorisant sa dégradation par le lysosome. [...] / Alzheimer’s disease (AD) is the most common neurodegenerative disorder of the old age, characterized by the presence of two major neuropathological features : neurofibrillary tangles and senile plaques. These plaques are composed of the A&#946; peptides cleavage product of the amyloid precursor protein (APP). Proteolytic processing of APP is modulated by the action of enzymes &#945;-, &#946;- and &#947;-secretases with the latter two mediating the amyloidogenic pathway. Suggesting that processing of APP is a key step in the pathology of AD. However, even if extensively studied, this APP metabolism is still not fully characterized. With this background, we postulate that the characterization of new actors of the APP metabolism might help for a more subtle understanding of this APP metabolism and trafficking. We focused on the ADAMs and related proteins with the hypothesis that ADAMs and related proteins, under- or over-expressed in the brain of AD cases compared with the one of controls, may be of particular interest. Beyond the obvious implication of several ADAMs as &#945;-secretases, this hypothesis was also driven by several observations : (i) ADAMs have been involved in numerous biological processes including brain development, plasticity and repair as APP; (ii) several metalloproteases (MMP-2, -3 and -9) have been described to degrade A&#946; peptides. Using microarray to screen the expression of 117 ADAMs and MMPs was analyzed using total RNA extracted from cerebral tissue of 12 AD cases and 12 controls. We observed that 4 ADAMs were differentially expressed. We first confirmed that the ADAM30 expression was decreased in AD brains and we observed that ADAM30 under-expression was correlated with an increase in A&#946;42 deposition in AD brains. Consistently, over-expression of ADAM30 led to decrease APP metabolism and as a consequence, A&#946; secretion in two different cell lines (Moreover, under-expressed ADAM30 increases APP processing and A&#946; generation). This modification of the APP metabolism was directly linked to the ADAM30 catalytic properties. Our data suggest that catalytic activity of ADAM30 takes an important place in APP processing in a lysosome dependent manner and AD pathophysiological process.

Un enjeu majeur de la génétique évolutive est de comprendre comment l'adaptation locale se développe en population naturelle, et comment les différentes forces évolutives y contribuent. Les études expérimentales d'adaptation locale utilisent couramment les gradients altitudinaux présentant une variation spatiale marquée des conditions environnementales. Dans ces conditions, on s'attend à ce que la différentiation génétique pour les caractères (traditionnellement mesurée par QST) et pour les gènes déterminant ces caractères (traditionnellement mesurée par FSTq) le long du gradient soit gouvernée de façon prédominante par la sélection et les flux de gènes, et peu influencée en revanche par la dérive génétique et la mutation. En particulier, des études théoriques ont montré un découplage entre QST et FST lorsque que les flux de gènes sont forts et/ou que la sélection est récente. Dans cette étude, nous avons testé cette hypothèse en combinant une approche de génomique expérimentale et des simulations dans des populations naturelles de hêtre commun (F. sylvatica) séparées de ~trois kilomètres et soumis à des environnements contrastés.Pour l'approche expérimentale, nous avons échantillonné 4 populations sur deux gradients altitudinaux sur le Mont Ventoux (avec une population à haute altitude et une à basse altitude sur chaque gradient). Cinquante huit gènes potentiellement impliqué dans la réponse aux stress abiotiques et dans le débourrement ont été séquencés sur un total de quatre-vingt seize individus, révélant 581 SNPs (Single Nucleotide Polymorphisms). Différentes approches ont été utilisées pour identifier les SNP outlier, présentant une différentiation plus forte qu'attendu sous un modèle neutre sans sélection. Le nombre de SNPs outlier identifié comme étant sous sélection s'est révélé être grandement dépendant de la méthode utilisé. La méthode fréquentiste a détecté de nombreux outliers alors que l'approche bayésienne n'a pu permettre de détecter des SNPs sous sélection. Par ailleurs, nous avons utilisé un modèle mécaniste individu-centré pour simuler les patrons de diversité phénotypique et génétique attendus le long du gradient pour la phénologie du débourrement végétatif, un caractère généralement adaptatif dans la réponse aux variations de température. Les résultats des simulations confirment que la différentiation génétique observée pour le caractère (QST) est généralement plus forte que celle observée au gène (FSTq), et que cette différentiation génétique au trait intervient dès la première génération. Toutefois, les tests d'outlier conduits sur le le modèle simulé ont révélé que plus de 95% des SNPs outlier sont des faux positifs. Comme dans l'approche expérimentale, l'approche Bayésienne ne s'est pas révélé suffisamment fiable pour détecter des QTLs dans des populations spatialement proche et génétiquement faiblement différentiée. Néanmoins une approche multi-locus basée sur un estimateur peu utilisé en génétique (le Zg) a révélé la forte corrélation inter-populations inter-gènes des QTLs confirmant les attendus théoriques. Toutefois, cette approche ne permet pas de détecter précisément les QTLs sans connaissance a priori sur les QTLs. En conclusion, les travaux de cette thèse mettent en évidence la rapidité des changements génétique qui interviennent en moins de 5 générations pendant la modification du climat, et la difficulté de détecter les gènes codant pour des traits complexes. / A major challenge in population genetics is to understand the local adaptation process in natural population and so to disentangle the various evolution forces contributing to local adaptation. The experimental studies on local adaption generally resort to altitudinal gradients that are characterized by strong environmental changes across short spatial scales. Under such condition, the genetic differentiation of the functional trait (measured by the Qst) as well as the genes coding for trait (measured by Fstq) are expected to be mainly driven by selection and gene flow. Genetic drift and mutation are expected to have minor effect. Theoretic studies showed a decoupling between Qst and Fst under strong gene flow and / or recent selection. In this study, I tested this hypothesis by combining experimental and modelling genomic approach in natural population of Fagus sylvatica separated by ~3 kilometres and under contrasted environments.Sampling was conducted in south-eastern France, a region known to have been recently colonised by F.sylvatica. Four naturally-originated populations were sampled at both high and low elevations along two altitudinal gradients. Populations along the altitudinal gradients are expected to be subjected to contrasting climatic conditions. Fifty eight candidate genes were chosen from a databank of 35,000 ESTs according to their putative functional roles in response to drought, cold stress and leaf phenology and sequenced for 96 individuals from four populations that revealed 581 SNPs. Classical tests of departure of site frequency spectra from expectation and outlier detection tests that accounted for the complex demographic history of the populations were used. In contrast with the mono-locus tests, an approach for detecting selection at the multi-locus scale have been tested.The results from experimental approaches were highly contrasted according the method highlighting the limits of those method for population loosely differentiated and spatially close. The modelling approach confirmed the results from the experimental data but revealed that up to 95% of the SNPs detected as outliers were false positive. The multi-locus approach revealed that the markers coding for the trait are differentially correlated compared to the neutral SNPs. But this approach failed to detect accurately the markers coding for the trait if no a priori knowledge is known about them. The modelling approach revealed that genetic changes may occur across very few generation. But while this genetic adaptation is measurable at the trait level, the available method for detecting genetic adaptation at the molecular level appeared to be greatly inaccurate. However, the multi-locus approach provided much more promise for understanding the genetic basis of local adaptation from standing genetic variation of forest trees in response to climate change.

5 Abstract This diploma thesis titled as Attraction and recruitment of university graduates: different perspectives of interested parties discusses the possible implementation of graduate recruitment within different organization. The text is divided into a theoretical part, which outlines the basic terminology and describes human resource activities connected with graduate recruitment and possible psychological profile and characteristics of graduates. The following chapters discuss situations that affect the perception of graduates in the labour market. The last chapter describes the available studies and investigations related to the evaluation of processes by participants. The empirical part focuses on the recruitment and selection of graduates in five different companies. The qualitative approach was used and semi-structured interviews were held with recruiters and graduates. The empirical part includes a comparison of processes in different companies, evaluate websites in the context of graduate recruitment and discuss the aspects of these processes, such as the perceived length of selection procedures, salary expectations of graduates and among other findings, what was most important to graduates when considering job offers. Keywords: Recruitment process, graduate, perception, candidate experience,...

As alterações cromossômicas estruturais associadas a fenótipos clínicos oferecem a oportunidade de identificação e localização de genes cujas mutações possam estar determinando essas patologias, tendo em vista a possibilidade de que esses genes podem ter sido alterados pelas quebras ou ter o número de cópias modificado. Um número cada vez maior de evidências aponta para a participação de certas seqüências do genoma na formação de rearranjos cromossômicos recorrentes e não recorrentes. Este trabalho compreendeu o estudo de duas translocações cromossômicas aparentemente equilibradas e uma duplicação do braço curto do cromossomo 20 em decorrência de mosaicismo materno. O objetivo foi determinar os pontos de quebra por hibridação in situ fluorescente (FISH) e identificar genes candidatos, alterados pelas quebras dos rearranjos e que pudessem explicar o quadro clínico dos portadores. A caracterização das seqüências nos pontos de quebra e a junção desses rearranjos é fundamental para a compreensão dos mecanismos de formação das alterações cromossômicas. A delimitação precisa dos segmentos deletados é necessária para a correlação com o quadro clínico. / Two apparently \"de novo\" balanced translocations and one duplication of the short arm of chromosome 20 were studied. Our aim was to determine the breakpoints by chromosomal analysis through fluorescentin situ hybridization (FISH) and identify candidate genes and how they were involved with the clinical phenotypes of the patients. Patient 1 carried a duplication of the short arm of chromosome 20 (p11.22p13), inherited from the mother that showed normal and dup(20) lymphocytes. The duplication was determined by FISH using BAC and PAC clones, and nine clones were duplicated except one (20p11.21). The patient shared many of the common characteristics of trisomy 20p including delay in motor development, hypertelorism, poor coordination, round face with prominent cheeks, vertebral and dental abnormalities and cranial asymmetry with high and large forehead. She also had learning difficulties, behavioral disorders and pubertal growth spurt at 12 years. As our patient is an example of pure trisomy 20p, the features are of particular importance to delineate the syndrome. Three genes were mapped on the segment that contain the duplication (20p11.2-13), one of these genes is the SSTR4 (Somatostatin receptor 4). The somatostatin is widely distributed throughout the body and is important regulator of endocrine and nervous system function. It is an inhibitor of growth hormone secretion. The second gene is the BMP2 that produce bone morphogenetic proteins and it has a direct function with the nervous system. The third gene is the GHRH that produce proteins connected with the growth hormone. These genes might have been over expressed and thus contributing to the patient\'s clinical features. Patient 2, carried a 46,XY,t(5;14)(q14.1;q31.3)de novo translocation. On chromosome 14 the breakpoint was mapped to a segment contained in BAC RP11-315O17 (14q31.3). On the chromosome 5 the breakpoint was mapped to a segment contained in BAC RP11-30D15 (5q14.1). Although the breakpoint, on the chromosome 14, has been mapped in 14q31.3, our patient shared many of the common characteristics of terminal 14q32 deletion: mental retardation, dolicocephaly, prominent ears, hypertelorism, strabismus, upturned palpebral fissures, highly arched palate, simian crease, severe myopia, coloboma and palpebral ptosis. As mental retardation and ocular abnormalities were the main patient\'s clinical features, we are suggesting that: 1) a region of segment 14q31.3 was deleted. 2) A gene inside this segment (14q31.3) could be responsible for ocular development and 3) a disrupted gene could interfere on the expression of other genes. On chromosome 5 eleven genes were localized and four of them are expressed in nervous system (AP3B1; SCAMP1; BHMT2 e CMYA5). One of these genes might have been disrupted and is contributing to the patient\'s clinical features. Patient 3 was the carrier of a 46,XY,t(1;15)(p13.2;q25.2)de novo translocation. The breakpoint on chromosome 15 was mapped to the segment contained in clone RP11-152F13 (15q25.2). The breakpoint on chromosome 1 was mapped to the segment contained in clone RP5-1037B23 (1p13.2). The genes mapped at the breakpoint regions of chromosome 1 and chromosome 15 are expressed in nervous system and muscles. Our patient shows few clinical features: speech delay, stutter and learning difficulties, probably because one or more of these genes, mapped at the breakpoint region, could be disrupted.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-01-25T15:40:39Z No. of bitstreams: 1 2015 - ?rica Cristina Xisto da Costa.pdf: 2608729 bytes, checksum: a906abcd8f725dc6509fedcf8de9e08c (MD5) / Made available in DSpace on 2017-01-25T15:40:39Z (GMT). No. of bitstreams: 1 2015 - ?rica Cristina Xisto da Costa.pdf: 2608729 bytes, checksum: a906abcd8f725dc6509fedcf8de9e08c (MD5) Previous issue date: 2015-07-31 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The gene encoding myostatin (MSTN), located on chromosome 18 (ECA 18) and the SNP BIEC2-808543 located in the intergenic region, which precedes the gene encoding the protein similar to the nuclear corepressor receptor dependent binder (LCORL), located on chromosome 3 (ACE 3) in horses, both positioned in regions that are associated with conformational traits of these animals. In view of this, we aimed to identify if the variations described in MSTN and LCORL loci exist in the study population; and to identify the effects of these polymorphisms on the growth profiles of these animals. For this purpose, the characteristics measured were weight, height at withers and hip height of foals at different ages, belonging to the Coudelaria e Campo de Instru??o de Rinc?o do Ex?rcito Brasileiro. Nonlinear mixed models were adjusted resulting from the combination of six nonlinear simple models, Brody (1945), Gompertz (Winsor, 1932), Logistics (Ratkowski, 1983), Von Bertalanffy (1957), generalized Michaelis- Menten (Lopez et al., 2000) and Richards (1959) associated with four types of variance functions for each model, homogeneous, exponential, asymptotic and staggered. The polymorphism described in the promoter region of the MSTN gene was not found in the studied population, in which there has been only the T allele, however the BIEC2-808543 polymorphism, located in the region prior to the LCORL gene is significantly associated (P <0, 05) to the characteristics evaluated, in which the animals who presented the genotype TT were smaller and lighter when compared to the other genotypes. There was no significant difference between animals with CT and CC genotype. The model that best describes the growth curve for body mass variance is the model of Brody (1945) associated with the scaled variance for the variable height at the withers the model that best fit was the Von Bertalanffy (1957) (adjusted without polymorphism effect in b) parameter associated with the asymptotic variance and the characteristic hip height the model that best described was that of Brody (1945) associated with the asymptotic variance, explaining that the nonlinear mixed models are indeed promising to describe equine growth curves, for the simple models did not differ much among themselves what defined in fact the selection of the model was the variance, being for body mass staggered variance and the height at the withers and on the back, the asymptotic variance. This polymorphism can be used as molecular markers for early selection of foals as to the characteristics evaluated / O gene que codifica a miostatina (MSTN), localizado no cromossomo 18 (ECA 18) e o SNP BIEC2-808543 localizado na regi?o interg?nica que antecede o gene que codifica a prote?na semelhante a correpressor de receptor nuclear dependente de ligante (LCORL), localizado no cromossomo 3 (ECA 3) de cavalos, ambos posicionados em regi?es que est?o associadas ?s caracter?sticas conformacionais destes animais. Diante disto, objetivamos identificar se as varia??es descritas nos loci MSTN e LCORL, existem na popula??o em estudo; al?m de verificar os efeitos desses polimorfismos sobre os perfis de crescimento desses animais. Com este intuito foram mensuradas as caracter?sticas massa corporal, altura na cernelha e altura na garupa de potros em diversas faixas et?rias, pertencentes ? Coudelaria e Campo de Instru??o de Rinc?o do Ex?rcito Brasileiro. Foram ajustados modelos n?o lineares mistos que resultaram da combina??o de seis modelos n?o lineares simples, Brody (1945), Gompertz (Winsor, 1932), Log?stico (Ratkowski, 1983), Von Bertalanffy (1957), Michaelis-Menten generalizado (L?pez et al., 2000) e Richards (1959), associados a quatro tipos de fun??es de vari?ncia para cada modelo, homog?nea, exponencial, assint?tica e escalonada. O polimorfismo descrito na regi?o promotora do gene MSTN n?o foi encontrado na popula??o estudada, na qual observa-se apenas o alelo T, entretanto o polimorfismo BIEC2-808543, localizado na regi?o que antecede o gene LCORL, est? significativamente associado (P<0,05) ?s caracter?sticas avaliadas, sendo os animais que apresentaram o gen?tipo TT menores e mais leves quando comparado com os demais gen?tipos. N?o foi observada diferen?a significativa entre os animais com gen?tipo TC e CC. O modelo que melhor descreve a curva de crescimento para a vari?vel massa corporal ? o modelo de Brody (1945) associado com a vari?ncia escalonada, para a vari?vel altura na cernelha o modelo que melhor se ajustou foi o de Von Bertalanffy (1957) (ajustado sem efeito de polimorfismo no par?metro b) associado com a vari?ncia assint?tica e para a caracter?stica altura na garupa o modelo que melhor a descreveu foi o de Brody (1945) associado ? vari?ncia assint?tica, elucidando que os modelos n?o-lineares mistos s?o de fato promissores para a descri??o de curvas de crescimento de equinos, pois os modelos simples n?o diferiram muito entre si o que definiu de fato a sele??o do modelo foi a vari?ncia, sendo para massa corporal a vari?ncia escalonada e para as alturas, na cernelha e na garupa, a vari?ncia assint?tica. Este polimorfismo pode ser utilizado como marcador molecular para sele??o precoce de potros quanto ?s caracter?sticas avaliadas

Submitted by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2016-09-22T13:21:31Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) BARBRA KATYÚSCYA SANCHES MORAES.pdf: 1057225 bytes, checksum: 43106c269cbed3f116c11fc89abec2f5 (MD5) / Approved for entry into archive by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2016-09-22T13:22:21Z (GMT) No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) BARBRA KATYÚSCYA SANCHES MORAES.pdf: 1057225 bytes, checksum: 43106c269cbed3f116c11fc89abec2f5 (MD5) / Made available in DSpace on 2016-09-22T13:22:21Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) BARBRA KATYÚSCYA SANCHES MORAES.pdf: 1057225 bytes, checksum: 43106c269cbed3f116c11fc89abec2f5 (MD5) Previous issue date: 2015-05-29 / Neste estudo, um método de CLAE-DAD foi desenvolvido e validado para a determinação de LASSBio-1736 em plasma de rato usando diclofenaco de sódio como padrão interno (PI). Extração líquido-líquido com acetonitrila foi utilizado para extrair LASSBio-1736. A separação cromatográfica foi realizada com coluna Waters Spherisorb®S5 ODS2 C18 (150 mm x 4,6 mm, 5μm), fase móvel isocrática composta por Trietilamina 0,3% (pH 4), metanol e acetonitrila (45:15:40, v/v/v) com vazão de 1 mL/min. Ambos LASSBio-1736 e PI foram eluídos em 4,2 e 5 min, respectivamente. O limite inferior de quantificação foi de 0,2 μg/mL e linearidade entre 0,2 - 4 μg/mL, com um r2> 0,99. A exatidão do método foi > 90,5%. Os desvios padrão relativos intra e inter-dias foram < 6,19 e < 7,83%, respectivamente. O método mostrou sensibilidade, linearidade, precisão, exatidão e seletividade necessária para quantificar LASSBio-1736 em estudos farmacocinéticos pré-clínicos O presente trabalho também investigou a farmacocinética plasmática e a distribuição do LASSBio-1736 em ratos Wistar. A farmacocinética de LASSBio-1736 foi investigada após a administração de dose intravenosa (3,2 mg/kg), por via oral e intraperitoneal (12,6 mg/kg).. A distribuição nos tecidos foi avaliada após administração de dose i.v. bolus. Os resultados para a via intravenosa indicam longo tempo meia-vida (24,3 ± 8,2 h), depuração de 49,3 ± 9,8 mL/Kg*h e volume de distribuição de 1,16 ± 0,3 L/kg. Para a via oral o tempo meia-vida foi de 28,6 ± 4,6 h, depuração de 49,7 ± 13 mL/Kg*h e volume de distribuição de 1,47 ± 0,34 L/kgforam semelhantes e biodisponibilidade de 12%. Para a via intraperitoneal o tempo meia-vida foi de 26 ± 8,9 h, depuração de 58 ± 19,6 mL/Kg*h e volume de distribuição de 1,8 ± 0,8 L/kg e biodisponibilidade de 38%. O LASSBio-1736 demonstrou penetração tecidual adequada no fígado, baço e pele. Com base nas características farmacocinéticas de outros fármacos leishmanicidas e a proposição de decisão para estudos farmacocinéticos visando a descoberta de candidatos a fármacos e a continuidade dos estudos, o LASSBio-1736 possui características farmacocinéticas apropriadas para um medicamento leishmanicida e novos estudos devem ser realizados para o escalonamento interespécies, além de estudos farmacológicos e toxicológicos complementares. / In this study, a method was developed and validated for HPLC-PDA determination LASSBio-1736 in rat plasma using diclofenac sodium as internal standard (IS). Liquid-liquid extraction was used to extract acetonitrile LASSBio-1736. The chromatographic separation was performed with Waters Spherisorb®S5 ODS2 C18 column (150 mm x 4.6 mm, 5μm), isocratic mobile phase consisting of Triethylamine 0.3% (pH 4), methanol and acetonitrile (45:15:40, v / v / v) with a flow rate of 1 mL/min. Both LASSBio-1736 and IS were eluted at 4.2 and 5 min, respectively. The lower limit of quantification was 0.2 μg/mL and linearity between 0.2 - 4 μg/mL, with r2> 0.99. The accuracy was > 90.5%. The relative standard deviation within and between days were < 6.19 and < 7.83%, respectively. The method showed sensitivity, linearity, precision, accuracy and selectivity needed to quantify LASSBio-1736 in preclinical pharmacokinetic studies. This study also investigated the plasma pharmacokinetics and distribution of LASSBio-1736 in Wistar rats. The pharmacokinetic LASSBio-1736 was investigated after intravenous dose administration (3.2 mg/kg) intraperitoneally and orally (12.6 mg/kg). The tissue distribution was evaluated after iv bolus dose administration. The results indicated an intravenous long half-life (24.3 ± 8.2 h) clearance 49.3 ± 9.8 mL/kg*h and the volume of distribution 1.16 ± 0.3 L/ kg. The oral route the half-life was 28.6 ± 4.6 h, clearance 49.7 ± 13 mL/kg*h and volume of distribution 1.47 ± 0.34 L/kg and bioavailability of 12%. The intraperitoneally half-life was 26 ± 8.9 h, clearance 58 ± 19.6 mL/kg*h and the volume of distribution 1.8 ± 0.8 L/kg and bioavailability of 38%were similar. The LASSBio-1736 showed adequate tissue penetration for liver, spleen and skin. Based on the pharmacokinetic characteristics of other antileishmanial drugs and the decision proposition for pharmacokinetic studies for the discovery of drug candidates and continuing studies, the LASSBio-1736 has pharmacokinetic characteristics appropriate for a leishmanicide and new drug studies should be conducted to the interspecies scaling, and additional pharmacological and toxicological studies.

Online education is becoming more prominent, but it has been found that employers are biased against hiring job applicants with online education. The influence of mode of education and degree level on employer perceptions of applicants’ hireability was investigated. It was hypothesized that employer perceptions would decrease as the education of an applicant moved to a category consisting of more online education. It was hypothesized that employers would be less likely to move forward with applicants in the hiring process (viability) as applicants’ education moved to a category consisting of more online education. It was hypothesized that degree level would moderate the relationship of mode of education on employer perceptions of applicants’ hireability and viability. Although it was expected that perceptions of hireability would increase as the applicants’ degree level increased and that perceptions would decrease as applicants’ education moved to a category consisting of more online education, as mode of education moved from hybrid to online, employer perceptions of applicants’ hireability were expected to decrease as applicants’ college degree level advanced. Survey results were collected using Qualtrics Resume Screener service. Results showed employer biases against an applicant’s hireability and viability exist as an applicants’ education moved to a category consisting of more online education, but degree level did not change this relationship. Theories were applied to results to provide possible explanations for the biases. The limitations of the current study as well as the theoretical and practical implications of the results are discussed.