The recent discovery of Cr4AlB4, a laminated ternary metal boride belonging to the family of layered MAB-phases, where the transition metal boride layers are interleaved by an A layer, has spurred theoretical investigation for novel M4AlB4 phases. In this study, first-principles calculations were applied in order to investigate the thermodynamical stability and mechanical properties of M4AB4 where M = Cr, Hf, Mo, Nb, Ta, Ti, V, W, Zr while the A layer was kept fixed as Al. The thermodynamical stability calculations validate the recently discovered Cr4AlB4 phase’s stability and suggest the neighboring phase Mo4AlB4 to be stable. Additionally, the phases Mo3AlB4, Mo2AlB2, Ti4AlB4 and Ti2AlB2 indicates phases close to stable with a formation enthalpy within the range of 0 < ∆H < 25 meV per atom compared to competing phases. Hence dynamical stability investigations were carried out, which indicates Mo4AlB4 to be dynamically stable. The stability of Mo4AlB4 does encourage synthesizing attempts to be kept in mind as a future project. Phase stability trends of the 111, 212, 314 and 414 compositions were discovered, where a 212, 314 and 414 composition is seen to be more stable for an M-element with lower electron configuration. Furthermore, the mechanical properties of the 414 compositions were investigated by systematically straining the unit cell in different directions. The bulk-, shear- and Young’s-modulus were derived and are presented, where Ti4AlB4 demonstrates values similar to the commended Ti2AlC MAX-phase. Finally, ductility plots are presented which purposes a linear trend between the elements of group IV, V and VI. Based on the results, further studies with a focus on the temperature and magnetization’s impact on the stability and mechanical properties are suggested.

Sex determination is a key developmental process in several species regulated by sexrelated transcription factors. In many species a gene called doublesex and mab-3 related transcription factor 1 (dmrt1), plays an important role in sexual differentiation. I used African clawed frogs (Xenopus) to examine function of dmrt1 in two species: a diploid species, X. tropicalis, and an allotetraploid species, X. laevis. In both species, dmrt1 is an autosomal gene; Xenopus tropicalis has one copy of dmrt1 and X. laevis has two homeologous copies that by definition are derived from whole genome duplication: dmrt1.L and dmrt1.S in X. laevis. We generated knockouts of each of these genes to further examine their function in sexual differentiation. Histological examination showed testicular dysgenesis in X. tropicalis dmrt1 and X. laevis dmrt1.L null males whereas dmrt1.S null males presented no obvious difference in sperm density compared to wildtype males. X. tropicalis dmrt1 and X. laevis dmrt1.L null females were found to completely lack reproductive organs and are infertile whereas dmrt1.S null females appeared unaffected. The contrasting results between dmrt1.L and dmrt1.S in X. laevis provides evidence of both neofunctionalization and subfunctionalization following gene duplication and suggest that gene duplication is a major contributor to evolutionary change. Additional investigation of the transcriptome of these frogs and the role of dmrt1 in the secondary sex characteristic vocalization provides further evidence of the role of dmrt1 in development. Comprehensively, this investigation provides further knowledge of the role of dmrt1 and homeologs of this gene in sexual differentiation and introduces a novel aspect of this gene in female development. Future efforts are focused on generating double knockouts for dmrt1.L and dmrt1.S, further examining the role of dmrt1.S in somatic cell function and developing additional mutant lines in other Xenopus for comparative analysis. / Thesis / Master of Science (MSc) / In many species sexual differentiation is a crucial developmental event. Surprisingly, however, the systems orchestrating sexual differentiation are highly variable among species. The doublesex and mab-3 related transcription factor 1 (dmrt1) gene plays a role in sexual differentiation in many groups, but its specific roles in this process are incompletely characterized and potentially diverse. We used genetic engineering in two species of African clawed frog (Xenopus) to disable function of dmrt1 in order to explore effects on gonadal development and the development of secondary sex characteristics. We found that dmrt1 is required for normal ovary or testis development in both Xenopus species, and that functional divergence occurred following duplication of dmrt1 by whole genome duplication. Taken together, these findings identify previously uncharacterized roles of dmrt1 in Xenopus and provide evidence of dynamic functional evolution of this important gene.

Flera kommuner runt om i Sverige rapporterar om att man har bostadsbrist. Orsaker tillatt det finns en bostadsbrist i Sverige är dels den höga byggkostnaden delsbefolkningsökning som sker. För att kunna möta upp bristen samt ökningen har detproducerats bostäder i en stor volym. Samtidigt som man bygger en stor mängd bostäder,går det att ifrågasätta hur många av dessa som håller viktiga kvaliteter som krävs förbostäder ska kunna uppnå trivsel och hemkänsla.Bostadsbristen som vi befinner oss i nu kan liknas med den bostadsbrist som Sverigebefann sig i på 50- och 60-talet. Det ledde då till att miljonprogrammet startades undermitten av 60-talet till mitten av 70-talet. Då byggdes mycket på kort tid och därefterupptäcktes brister i kvalitén och utförande. För att undvika bristerna i kvalitet är detviktigt att beakta grundfaktorer som skapar trivsel. Några av dessa grundfaktorer är de“omätbara värdena” som i samverkan med varandra skapar hemkänsla och trivsel i enbostad.Att skapa trivsamma bostäder och byggnader är en väldigt komplex uppgift i form av attman kan missa vissa grundfaktorer som främjar trivsel i bostäder. Därför behövs studiergöras för att se om dessa grundfaktorer finns med i dagens nyproduktion av bostäder.Detta på grund av att dagens byggindustri kan glömma viktiga punkter i sin projekteringoch bara tänker på kostnader och tid. Vilket gör att många viktiga parametrar kanglömmas bort och därför skapas en mindre trivsam bostad.Syftet med arbetet är att undersöka planlösningar för att kunna få en insikt i hur kvaliteteni nya lägenheter i flerbostadshus är. Arbetet granskar planlösningar från fyra kommunersom är slumpmässigt utvalda. För att avgöra kvaliteten i bostäderna utgår arbetet frånboken “Bostadens omätbara värden” av Ola Nylander samt “Manual för analys avBostadskvalitet” som är framtagen av forskare vid Chalmers.Den valda metoden är en fallstudie av stickprovskaraktär. I studien har de utvaldaplanlösningarna granskats och analyserats baserat på fyra parametrar om omätbara värdensamt en parameter med mätbart värde. De fyra omätbara värdena är rundgång, axialitet,dagsljus och rumsorganisation. Det mätbara värdet är Potential för kvarboende.Resultatet av arbetet redovisas genom figurer på planlösningarna där tydligt visas de olikaparametrar i figurer. Gruppen har gått igenom olika bostadsstorlekar från de fyra olikakommunerna. Storlekarna på bostäderna är 2 ROK, 3 ROK och 4 ROK.Slutsatsen av studien är att nya bostäder som byggs överlag uppfyller kraven uppställda i”Bostadens omätbara värden” samt ”Manual för analys av Bostadskvalitet”. Vidprojektering av bostäderna visas det i denna studie att arkitekter och projektörer har tänktpå de omätbara och mätbara värdena när lägenheterna tagits fram. Storleken på bostadenhar stor betydelse, eftersom det går att få med flera och bättre parametrar i bostaden omde innehåller flera rum. I mindre bostäder finns i vissa fall, brister i antal parametrar somsänker kvaliteten. Parametern från Manual för analys av Bostadskvalitét (MAB),Potential för kvarboende, gick att finna i planlösningarna på ett tydligt sätt. / Several municipalities around Sweden report that they have a housing shortage. Reasonswhy there is a housing shortage in Sweden are partly the high construction costs andpartly the population increase that is taking place. In order to meet the shortage as well asthe increase, housing has been produced in a large volume. At the same time as a largeamount of housing is being built, it is possible to question how many of these meet theimportant qualities required for housing to be able to achieve comfort and a sense offeeling of home.The housing shortage that we are in now can be compared to the housing shortage thatSweden was in in the 50s and 60s. That then led to the million program being started inthe mid-60s to the mid-70s. Back then, a lot was built in a short time and then flaws in thequality and execution were discovered. In order to avoid deficiencies in quality, it isimportant to consider basic factors that create well-being. Some of these basic factors arethe "immeasurable values" which, in cooperation with each other, create a sense of homeand well-being in a home.Creating pleasant homes and buildings is a very complex task and therefore it is easy tomiss certain basic factors that promote well-being in homes. Therefore, studies need to bedone to see if these basic factors are included in today's new housing production. This isbecause today's construction industry can forget important points in its planning and onlythinks about costs and time. Which means that many important parameters can beforgotten and therefore create a less pleasant home.The purpose of the work is to investigate floor plans in order to gain an insight into thequality of new apartments in apartment buildings. The work examines planning solutionsfrom four municipalities that are randomly selected. In order to determine the quality ofthe housing, the work is based on the book "The home ́s immeasurable value" by OlaNylander and "Manual for analysis of housing quality" developed by research scientist atChalmers.The chosen method is a random sample study. In the study, the selected floor plans havebeen reviewed and analyzed based on four parameters of immeasurable values and oneparameter with measurable value. The four immeasurable values are movement, axiality,daylight and room organization. The measurable value is Potential for remainingresidents.The result of the work is shown through figures on the floor plans where the variousparameters are clearly shown in figures. The group has gone through different housingsizes from the four different municipalities. The sizes of the homes are 2 RAK, 3 RAKand 4 RAK.The conclusion of the study is that new homes that are built generally meet therequirements set out in "The immeasurable values of housing" and "Manual for analysisof housing quality". When designing the homes, it is shown in this study that architectsand designers have thought about the immeasurable and measurable values when theapartments were designed. The size of the home is of great importance, as it is possible toget more and better parameters in the home if they contain several rooms. In smallerhomes there are, in some cases, deficiencies in the number of parameters that lower thequality. The parameter from the Manual for Analysis of Housing Quality (MAB),Potential for remaining residents, could be found in the floor plans in a clear way.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

vi ABSTRACT IL-2 has been used in cancer therapy and also for other applications like treatment of chronic viral infections or as an adjuvant for vaccines. However, treatment with IL-2 is rather difficult due to its severe side effects. These toxicities, associated with high-dose treatment necessary for IL-2 to function, have been found the most limiting factor for IL- 2 applications. Further, particular anti-IL-2 monoclonal antibodies (mAb) can actually increase biological activity of IL-2 rather than block it. Binding of IL-2 to anti-IL-2 mAb creates a superagonistic immunocomplexes which have dramatically higher and selective biological activity in comparison to free IL-2 in vivo. Such approach may finally over- come the difficulties associated with administration of IL-2, thus opening brand new scopes for IL-2 and its application not only in the field of tumor therapy. We have shown that IL-2 immunocomplexes composed of IL-2 and anti-IL-2 mAb S4B6 (IL-2/S4B6) stimulate predominantly cells expressing CD122 and CD132 (dimeric IL-2 receptor), i.e. NK and MP CD8+ T cells, with Treg,  T and NKT cells being expanded as well. IL-2/S4B6 are able to drive the expansion of activated naive CD8+ T cells into functional memory-like CD8+ T cells. Moreover, these immunocomplexes exert therapeu- tical potential alone...

In this thesis, we address several generic problems concerned with procurement of tasks from a crowd that consists of strategic workers with uncertainty in their qualities. These problems assume importance as the quality of services in a service marketplace is known to degrade when there is (unchecked) information asymmetry pertaining to quality. Moreover, crowdsourcing is increasingly being used for a wide variety of tasks these days since it offers high levels of flexibility to workers as well as employers. We seek to address the issue of quality uncertainty in crowdsourcing through mechanism design and machine learning. As the interactions in web-based crowdsourcing platform are logged, the data captured could be used to learn unknown parameters such as qualities of individual crowd workers. Further, many of these platforms invite bids by crowd workers for available tasks but the strategic workers may not bid truthfully. This warrants the use of mechanism design to induce truthful bidding. There ensues a complex interplay between machine learning and mechanism design, leading to interesting technical challenges. We resolve some generic challenges in the context of the following problems. Design of a quality eliciting mechanism with interdependent values We consider an expert sourcing problem, where a planner seeks opinions from a pool of experts. Execution of the task at an assured quality level in a cost effective manner turns out to be a mechanism design problem when the individual qualities are private information of the experts. Also, the task execution problem involves interdependent values, where truthfulness and efficiency cannot be achieved in an unrestricted setting due to an impossibility result. We propose a novel mechanism that exploits the special structure of the problem and guarantees allocative efficiency, ex-post incentive compatibility and strict budget balance for the mechanism, and ex-post individual rationality for the experts. Design of an optimal dimensional crowdsourcing auction We study the problem faced by an auctioneer who gains stochastic rewards by procuring multiple units of a service from a pool of heterogeneous strategic workers. The reward obtained depends on the inherent quality of the worker; the worker’s quality is fixed but unknown. The costs and capacities are private information of the workers. The auctioneer is required to elicit costs and capacities (making the mechanism design dimensional) and further, has to learn the qualities of the workers as well, to enable utility maximization. To solve this problem, we design a dimensional multi-armed bandit auction that maximizes the expected utility of the auctioneer subject to incentive compatibility and individual rationality while simultaneously learning the unknown qualities of the agents. Design of a multi-parameter learning mechanism for crowdsourcing We investigate the problem of allocating divisible jobs, arriving online, to workers in a crowd-sourcing platform. Each job is split into a certain number of tasks that are then allocated to workers. These tasks have to meet several constraints that depend on the worker performance. The performance of each worker in turn is characterized by several intrinsic stochastic parameters. In particular, we study a problem where each arriving job has to be completed within a deadline and each task has to be completed, honouring a lower bound on quality. The job completion time and quality of each worker are stochastic with fixed but unknown means. We propose a learning mechanism to elicit the costs truthfully while simultaneously learning the stochastic parameters. Our proposed mechanism is dominant strategy incentive compatible and ex-post individually rational with asymptotically optimal regret performance.

Philosophiae Doctor - PhD / The domesticated apple (Malus x domestica Borkh.) has been distributed into diverse climatic conditions worldwide for commercial production of fruit. Apple trees need exposure to cold temperatures, referred to as chill unit (CU) accumulation during winter, in order for budbreak to occur promptly and uniformly after winter. In warmer production areas the application of dormancy breaking chemicals has enabled successful production of high chilling requiring apple cultivars in suboptimal environmental conditions. In the Western Cape region of South Africa it is common orchard practice to apply dormancy breaking chemicals after winter in order to stimulate vegetative growth. If this is not done prolonged dormancy symptoms (PDS) are experienced which include extended rest, less synchronised breaking of buds and reduced branching. An increasing awareness of both global warming and the negative effects associated with the use of chemical sprays (for both pest and disease resistance and growth regulation) has resulted in the need to breed cultivars better adapted to current and future environmental conditions. The breeding of new cultivars using conventional breeding methods is a time consuming process, especially in perennial tree species with a long juvenile phase such as apple. The implementation of marker-assistedbreeding (MAB) and selection (MAS) will enable the selection of favourable genotypes at a very early seedling stage. Although markers linked to genes involved in disease resistance for a variety of known apple pathogens have been identified and are already in use in breeding programs, the genetic determinants of dormancy related characteristics residing within the bud itself iii (endodormancy) are poorly understood. This hampers the genetic improvement of such characters. Although this study focused on time of initial vegetative budbreak IVB, there are various other characteristics that can be associated with dormancy, such as position and number of budbreak and budbreak duration.

La connaissance des effets et modes d’action des QTL (Quantitative Trait Loci) de résistance des plantes aux maladies est un enjeu majeur pour l’exploitation durable des résistances génétiques en agriculture. Dans le pathosystème pois/Aphanomyces euteiches, présentant une importance économique majeure, l’objectif de la thèse a consisté à valider les effets de QTL de résistance, seuls ou en combinaison, et à connaitre leurs modes d’action sur les étapes du cycle de l’agent pathogène. Un total de 157 NILs (Near Isogenic Lines) de pois issues d’un programme de Back-cross Assisté par Marqueurs, porteuses d’aucun, un, deux ou trois des sept principaux QTL de résistance préalablement identifiés, a été génotypé à l’aide d’une puce 15K SNPs et évalué pour la résistance. Les NILs porteuses de QTL à effets majeurs, seuls ou en combinaison avec des QTL à effets mineurs,ont présenté des niveaux accrus de résistance partielle en conditions contrôlées et au champ par rapport aux NILs dépourvues de QTL, dans différents fonds génétiques. Certaines NILs comportant des QTL individuels ou combinés à effets mineurs ont également montré un niveau réduit de sévérité de la maladie dans l’une ou les deux conditions de test. La plupart des QTL a présenté des effets significatifs sur le ralentissement de l’infection et/ou de la quantité d’ADN pathogène ayant colonisé la racine pendant sept jours après inoculation. Cette thèse fournit des outils et éléments de choix de QTL à combiner en sélection pour augmenter l’efficacité de la résistance partielle à A. euteiches dans les futures variétés de pois / Knowledge of the effects and action modes of resistance QTL to plant diseases is a major challenge for the durable use of genetic resistances in agriculture. In the pea/Aphanomyces euteiches pathosystem, which has a major economic importance, the aim of this study was to validate the single or combined effects of main resistance QTL, and study their action modes on steps of the pathogen life cycle. A total of 157 pea NILs (Near Isogenic Lines) created by Marker-Assisted Back-crossing and carrying no, one, two or three of the seven main resistance QTL previously identified, was genotyped using a 15K SNPs array and evaluated for resistance. The NILs carrying major-effect QTL, individually or in combination with minor-effect QTL, had increased levels of partial resistance in controlledconditions and in the field compared to NILs without QTL, in different genetic backgrounds. Several NILs carrying single or multiple minor-effect QTL also showed reduced levels of disease severity in one or the two test conditions. Most of the QTL had significant effects on slowing down infection and/or pathogen DNA quantity which had colonized the root for seven days after inoculation. This study gives tools and information for the choice of resistance QTL to use in pyramiding breeding strategies for increasing partial levels of resistance to A. euteiches in future pea varieties.

Thèse réalisée dans le cadre d'une collaboration avec UCB Pharma et la région Wallonne s'inscrivant dans le cadre du projet SAS. Le but de ce projet était de développer des implants sous-cutanés imprimés en trois dimensions pour permettre une libération d’anticorps thérapeutique de manière prolongée au cours du temps. En effet, les thérapies disponibles sont souvent administrées par voie intraveineuse, ce qui peut réduire la compliance des patients dû à l’inconfort et à la fréquence de ces administrations. Les systèmes de délivrance, tels que des implants, peuvent limiter les fréquences d’administration grâce à l’insertion d’un dispositif qui libèrera le principe actif au cours du temps durant une période donnée. Les implants s’inscrivent comme une alternative aux microsphères qui sont également des dispositifs développés et investigués en vue de favoriser l’adhésion et la compliance des patients. L’avènement du 3D dans le milieu pharmaceutique a montré une certaine frénésie liée au développement de la médecine personnalisée et à l’innovation du procédé dans ce secteur. La sélection d’un matériau biocompatible et biorésorbable tel que le PLGA représente une véritable plus-value dans le développement d’implant. Etant donné que ces implants sont biodégradables, le retrait n’est pas à envisager, ce qui limite les désagréments du patient à un seul acte chirurgical lors de l’implantation. Au cours de ce travail, une approche pragmatique a d’abord été abordée sur les procédés d’extrusion à chaud et de l’impression 3D en utilisant un polymère couramment employé dans l’impression grand public, le PLA. L’investigation des paramètres d’impressions (température d’impression, epaisseur de couche et vitesse d’impression) et l’usage de divers plastifiants (la triacétine (TA), le polyethylène glycol 400 (PEG 400), le citrate de triéthyle (TEC) et l’acétyle citrate de triéthyle (ATEC)) pour faciliter les procédés à chaud et dans l’idée de réduire les températures d’extrusion et d’impression du matériau ont été évalués. Ces essais ont démontré l’effet de la température d’impression sur la qualité de l’impression et principalement sur les propriétés du matériau comme la force de traction et la ductilité. De plus, l’ajout de plastifiant à la matrice du PLA a permis de diminuer sa température de transition vitreuse. Par exemple, la température de transition vitreuse du PLA a été diminuée de 53 °C à 34 °C par l’ajout de PEG 400. Cette approche avait pour but d’évaluer la possibilité de diminuer les températures d’impression dans l’optique d’encapsuler à chaud un anticorps sensible à la chaleur pour la suite de ce travail.Ensuite, le développement de filaments imprimables contenant des anticorps a été abordé et mis en place à l’aide d’un modèle d’anticorps polyclonal disponible en grandes quantités et à des coûts relativement faibles. Un anticorps à l’état solide a été favorisé dans le procédé car il est largement accepté que les protéines sous forme solide sont plus stables au cours du temps en comparaison aux solutions d’anticorps. De plus, cet état solide facilite les manipulations précédant l’extrusion comme l’étape de mélange. Pour la réalisation des filaments, différents types de PLGA ont été investigués afin d’atteindre les propriétés nécessaires à l’impression en termes de diamètre mais également de comportement physique. Ces dérivés étaient caractérisés par des masses moléculaires différentes comme pour le PDLG5004 (44 kDa), le RG502 (7-17 kDa) et parmis eux, un copolymère PEG-PLGA (2 kDa-20 kDa). Un PLGA de faible masse moléculaire a été sélectionné pour développer ce filament. En effet, les extrusions étaient réalisables à une température maximum de 90 °C et les impressions à 113 °C minimum. L’un des enjeux cruciaux du développement de filament imprimable contenant un anticorps à haute concentration, au minimum 15% (w/w), était d’en assurer l’homogénéité. Cependant, l’usage de températures aussi élevées lors de l’impression a induit la dégradation de l’anticorps par la formation d’agrégats et principalement de fragments. Ces derniers sont généralement produits lors de procédé à haute température ou par l’usage de conditions drastiques telles que l’acidification du milieu. Cette plateforme a été adaptée à l’encapsulation d’anticorps thérapeutique fournit par UCB Pharma. L’usage d’un anticorps monoclonal possédant une stabilité supérieure à celle du modèle initialement utilisé permettrait d’identifier l’impact du procédé sur l’intégrité de l’anticorps. La formulation de l’anticorps a été réalisée en utilisant différents stabilisants conventionnels (sucrose (Suc), trehalose (Tre), 2-Hydroxypropyl-beta-cyclodextrine (HP-β-CD), inuline (Inu) et sorbitol (Sor)) et reconnus pour la stabilisation des protéines. A côté des excipients ajoutés, différentes quantités d’excipients ont été investigués. Ces manipulations ont montré que la stabilité de l’anticorps était privilégiée à l’aide du sucrose et du tréhalose à un ratio anticorps monoclonal:excipient de 2.0:1. En gardant ce ratio, l’ajout d’un acide aminé (leucine) aux deux disaccharides précédemment cités, a amélioré la stabilité de l’anticorps vis-à-vis des procédés à chaud (extrusion et impression 3D). L’homogénéité au sein des filaments imprimables et des pièces 3D a été confirmée tout au long du procédé. En effet, les charges en anticorps étaient similaires à la charge théorique de 15% (w/w). Aucune fragmentation de l’anticorps n’a été observée à l’issue des procédés à chaud. Cependant, une augmentation des agrégats de 2.6% en solution à 3.6% après impression 3D a été constatée à la fin du processus. Après avoir stabilisé l’anticorps, le but premier étant d’en promouvoir une libération prolongée au cours du temps. Les profils ont révélé une libération en trois phases au cours du temps mais avec un relargage après 24h relativement faible (< 5%) dû à la densité des matrices polymériques. Ensuite, la dégradation du polymère représente l’élément limitant la libération de l’anticorps au cours du temps. En effet, l’érosion du polymère joue un rôle clé dans la libération de l’anticorps encapsulé. La libération au cours du temps a été démontrée sur une période allant jusqu’à 15 semaines. La stabilité de l’anticorps dans le milieu de dissolution a été évaluée et une dégradation de celui-ci au cours du temps a été observée. Cette dégradation est principalement liée à l’érosion du polymère et à l’acidification du milieu au cours du test de dissolution. Après avoir optimisé la formulation de l’anticorps et avoir démontré la libération prolongée de celui-ci, son affinité restait à être étudiée. La capacité de l’anticorps à se lier à sa cible a pu être démontrée après 24h de dissolution mais cette affinité s’est réduite au cours de la durée de la dissolution avec une augmentation de l’agrégation et de la fragmentation de l’anticorps. Une étude de stabilité a également démontré que les implants imprimés en 3D sont stables à une température 5 °C sur une durée de 6 mois. Aucun élément de dégradation n’a été observé au cours du temps et l’affinité de l’anticorps a été préservée au cours de l’étude. Finalement, cette plateforme a également été évaluée pour l’encapsulation d’une troisième molécule biologique, un fragment d’anticorps monoclonal, pour d’une part en estimer la stabilité et l’applicabilité et d’autre part envisager une prochaine étude pré-clinique sur rongeurs. Le fragment d’anticorps a montré une stabilité supérieure à celle de l’anticorps monoclonal avec une faible agrégation après l’extrusion et l’impression. La libération prolongée du fragment a été évaluée sur 8 semaines et une libération du fragment de 79% a été observée avec une formulation contenant du tréhalose et de la leucine. En effet, les fragments d’anticorps ont une demi-vie plasmatique relativement faible, de l’ordre de 28 minutes, ce qui donne tout son sens à des systèmes à libération prolongée. Pour finir, la réalisation d’une étude pré-clinique permettrait de valider le modèle. En conclusion, ce travail a démontré la faisabilité de l’usage de l’impression 3D en vue de développer des systèmes à libération prolongée contenant des anticorps et en utilisant des procédés à hautes températures. Ces implants ont été caractérisés par une stabilité favorable et une libération intéressante qui feront l’objet d’investigation lors d’études pharmacocinétiques. / Doctorat en Sciences biomédicales et pharmaceutiques (Pharmacie) / info:eu-repo/semantics/nonPublished

Introdução: O gene DMRT1 é um fator muito importante, o qual induz a determinação sexual masculina. Estudos mais recentes têm demonstrado que o Dmrt1 possui um papel significante no desenvolvimento ovariano. Deleções restritas ao gene DMRT1 têm sido raramente identificadas em pacientes com disgenesia gonadal (DG) sem outras características sindrômicas. Objetivo: Pesquisar a presença de haploinsuficiência do gene DMRT1 (deleções e/ou mutações inativadoras) em um grupo grande de pacientes não sindrômicos com distúrbios do desenvolvimento sexual (DDS) por anormalidades gonadais. Polimorfismos do DMRT1, como fatores potenciais pelas anormalidades gonadais, foram também identificados. Pacientes e Métodos: Foram avaliados cerca de 39 pacientes portadores de DDS por anormalidades do desenvolvimento gonadal 46,XY: 24 com disgenesia gonadal parcial e 15 pacientes com disgenesia gonadal completa. As regiões codificadoras do DMRT1 e o domínio DM (exon 1) foram amplificados e sequenciados. A análise de Multiplex ligation probe amplification (MLPA) do DMRT1 foi realizada usando um kit comercial. Resultados: Deleção parcial ou total do DMRT1 não foi identificada pela técnica de MLPA. Oito variantes alélicas do DMRT1 foram identificados. Uma nova variante c.968-15insTTCTCTCT foi identificada em 6,4% e em 14,3% dos alelos dos pacientes 46,XY e indivíduos controles, respectivamente. Conclusão: Este estudo sugere que deleções parciais ou completas no DMRT1 e mutações inativadoras não são frequentemente encontradas em pacientes com anormalidades do desenvolvimento gonadal. Além disso, nenhuma das variantes alélicas identificadas neste grupo de pacientes poderia ser considerada como um marcador potencial polimórfico para disgenesia gonadal / Introduction Dmrt1 gene is a very important factor in inducing male sex determination, and more recently it has been demonstrated that Dmrt1 plays a significant role in ovary development. DMRT1 deletions have rarely been identified in patients with 46,XY gonadal dysgenesis (GD) without syndromic features. Objective- To screen for the presence of DMRT1 haploinsufficiency (deletions and/or inactivating mutations) in a large cohort of non-syndromic patients with disorder of sex development (DSD) due to abnormalities of gonadal development. DMRT1 polymorphisms, as potential susceptibility factors for gonadal abnormalities, were also investigated. Subjects and Methods- We evaluated 39 patients with 46,XY GD: 24 patients with the partial, and 15 with the complete form. The entire coding region (éxons 2-5) of DMRT1 and the DM domain (exon 1) were PCR-amplified and direct sequenced. Multiplex ligation probe amplification (MLPA) analysis of DMRT1 was carried out using a commercial kit. Results- Partial or total deletion of DMRT1 was not identified by MLPA technique. Eight allelic variants of DMRT1 were identified. The novel variant c.968-15insTTCTCTCT was identified in 6.4% and in 14.3% of the alleles of 46,XY patients and control subjects, respectively Conclusion- This study suggest that complete or partial DMRT1 deletions and inactivating mutations are not frequently found in patients with abnormalities of gonadal development. Additionally, none of the allelic variants identified in this cohort of patients could be considered a potential polymorphic susceptibility marker for gonadal dysgenesis