A Tissue-Engineered Microvascular System to Evaluate Vascular Progenitor Cells for Angiogenic Therapies

The ability of tissue engineered constructs to replace diseased or damaged organs is limited without the incorporation of a functional vascular system. To design microvasculature that recapitulates the vascular niche functions for each tissue in the body, we investigated the following hypotheses: (1) cocultures of human umbilical cord blood-derived endothelial progenitor cells (hCB-EPCs) with mural cells can produce the microenvironmental cues necessary to support physiological microvessel formation in vitro; (2) poly(ethylene glycol) (PEG) hydrogel systems can support 3D microvessel formation by hCB-EPCs in coculture with mural cells; (3) mesenchymal cells, derived from either umbilical cord blood (MPCs) or bone marrow (MSCs), can serve as mural cells upon coculture with hCB-EPCs. Coculture ratios between 0.2 (16,000 cells/cm2) and 0.6 (48,000 cells/cm2) of hCB-EPCs plated upon 3.3 µg/ml of fibronectin-coated tissue culture plastic with (80,000 cells/cm2) of human aortic smooth muscle cells (SMCs), results in robust microvessel structures observable for several weeks in vitro. Endothelial basal media (EBM-2, Lonza) with 9% v/v fetal bovine serum (FBS) could support viability of both hCB-EPCs and SMCs. Coculture spatial arrangement of hCB-EPCs and SMCs significantly affected network formation with mixed systems showing greater connectivity and increased solution levels of angiogenic cytokines than lamellar systems. We extended this model into a 3D system by encapsulation of a 1 to 1 ratio of hCB-EPC and SMCs (30,000 cells/µl) within hydrogels of PEG-conjugated RGDS adhesive peptide (3.5 mM) and PEG-conjugated protease sensitive peptide (6 mM). Robust hCB-EPC microvessels formed within the gel with invasion up to 150 µm depths and parameters of total tubule length (12 mm/mm2), branch points (127/mm2), and average tubule thickness (27 µm). 3D hCB-EPC microvessels showed quiescence of hCB-EPCs (<1% proliferating cells), lumen formation, expression of EC proteins connexin 32 and VE-cadherin, eNOS, basement membrane formation by collagen IV and laminin, and perivascular investment of PDGFR-β+/α-SMA+ cells. MPCs present in <15% of isolations displayed >98% expression for mural markers PDGFR-β, α-SMA, NG2 and supported hCB-EPC by day 14 of coculture with total tubule lengths near 12 mm/mm2. hCB-EPCs cocultured with MSCs underwent cell loss by day 10 with a 4-fold reduction in CD31/PECAM+ cells, in comparison to controls of hCB-EPCs in SMC coculture. Changing the coculture media to endothelial growth media (EBM-2 + 2% v/v FBS + EGM-2 supplement containing VEGF, FGF-2, EGF, hydrocortisone, IGF-1, ascorbic acid, and heparin), promoted stable hCB-EPC network formation in MSC cocultures over 2 weeks in vitro, with total segment length per image area of 9 mm/mm2. Taken together, these findings demonstrate a tissue engineered system that can be utilized to evaluate vascular progenitor cells for angiogenic therapies.

Altered pericyte-endothelial relations in the rat retina during aging: Implications for vessel stability

Mural cells (smooth muscle cells and pericytes) regulate blood flow and contribute to vessel stability. We examined whether mural cell changes accompany age-related alterations in the microvasculature of the central nervous system. The retinas of young adult and aged Wistar rats were subjected to immunohistofluorescence analysis of a-smooth muscle actin (SMA), caldesmon, calponin, desmin, and NG2 to identify mural cells. The vasculature was visualized by lectin histochemistry or perfusion of horse-radish peroxidase, and vessel walls were examined by electron microscopy. The early stage of aging was characterized by changes in peripheral retinal capillaries, including vessel broadening, thickening of the basement membrane, an altered length and orientation of desmin filaments in pericytes, a more widespread SMA distribution and changes in a subset of pre-arteriolar sphincters. In the later stages of aging, loss of capillary patency, aneurysms, distorted vessels, and foci of angiogenesis were apparent, especially in the peripheral deep vascular plexus. The capillary changes are consistent with impaired vascular autoregulation and may result in reduced pericyte-endothelial cell contact, destabilizing the capillaries and rendering them susceptible to angiogenic stimuli and endothelial cell loss as well as impairing the exchange of metabolites required for optimal neuronal function. This metabolic uncoupling leads to reactivation of

Los primeros pasos en el escorzo de la figura humana. "Un análisis desde la geometría"

A través de un recorrido histórico centrado en el Renacimiento, se exponen las aportaciones más significativas de teóricos y artistas en la evolución del escorzo de la figura humana desde un punto de vista geométrico. Los artistas que con sus dibujos y escritos ayudaban a otros en el aprendizaje de la representación, se apresuraron a incluir el estudio del escorzo en sus tratados, buscando métodos que facilitaran su dibujo sobre cualquier soporte y en cualquier posición en el espacio. En este artículo se analizan y enlazan los trazados propuestos por los estudiosos de la geometría, con las obras de arte que reflejan importantes avances en este sentido. Soportes como el lienzo, el muro o la bóveda, presentan al pintor superficies diferentes de trabajo y en distintas posiciones en el espacio. La figura humana, protagonista en la escena, tendrá que adaptarse a ellos para ser contemplada desde unos espacios arquitectónicos cada vez más amplios y con más posibilidades. Fue necesario dominar la perspectiva en la pintura, y muy particularmente en la mural, para ofrecer composiciones cada día más ambiciosas y sorprendentes.

Modulating mythology in a post-traumatic era: Murals and re-imaging in Northern Ireland.

Arguably the most ancient of the social media, wall paintings have been a persistent vehicle of cultural meaning management. The dynamics of myth markets are reflected in the sectarian murals of Northern Ireland. In this paper, we draw from consumer research literature on mythology and street art to explore the continuous revision of these wallscapes that seeks to address the enduring contradictions of civic ideology in contested political space. In particular, we focus on the use of classical, historical and pop-cultural mythologies to transform private space into public place. We examine the decommissioning of murals occurring in the wake of the Peace Accords, and speculate on the implications of the creation of a shared mythology for the future of mural painting and the state.

A iconografia de Santa Maria Madalena em Portugal até ao Concílio de Trento

Tese de mestrado Arte, Património e Teoria do Restauro, Universidade de Lisboa, Faculdade de Letras, 2012

A rentabilização pedagógica de um grupo fechado no Facebook na disciplina de inglês

Trabalho de projeto de mestrado, Educação (Área de Especialização em Educação e Tecnologias Digitais), Universidade de Lisboa, Instituto de Educação, 2014

Diálogos de modernidade no Azulejo em Portugal (1949 -1970)

Dissertação apresentada para cumprimento dos requisitos necessários à obtenção do grau de Mestre em História da Arte Contemporânea

DNA polymerase activity in trophoblast giant cells in the mouse /

In the developing mouse embryo, the diploid trophectoderm is known to undergo a diploid to giant cell transformation. These cells arise by a process of endoreduplication, characterized by replication of the entire genome without subsequent mitosis or cell division, leading to polyploidy and the formation of giant nuclei. Studies of 13.5 day rat trophoblast derived from the parietal yolk sac have indicated a relatively low rate of DNA polymerase a activity, the noinnal eukaryotic replicase, in comparison to that of DNA polymerase g. These results have suggested that endoreduplication in trophoblast giant cells may not employ the normal replicase enzyme, DNA polymerase a. In order to determine whether a 'switch' from DNA polymerase to DNA polymerase is a necessary concomitant of the diploid to giant cell transformation, two distinct populations of trophoblast giant cells, the primary giant cell derived from the mural trophectoderm and the secondary giant cell derived from the polar trophoectoderm were used. These two populations of trophoblast giant cells can be obtained from the tissue outgrowths of 3.5da blastocysts and the extraembryonic ectoderm (EX) and ectoplacental cone (EPC) of 7.5 day embryos respectively. Tissue outgrowths were treated with aphidicolin, a specific reversible inhibitor of eukaryotic DNA polymerase a, on various days after explantation. The effect of aphidicolin treatment was assessed both qualitatively, using autoradiography and quantitatively by scintillation counting and Feulgen staining. 3 DNA synthesis was measured in control and treated cultures after a Hthymidine pulse. Scintillation counts of the embryo proper revealed that DNA synthesis was consistently inhibited by greater than 907. in the presence of aphidicolin. Inhibition of DNA synthesis in the EX and EPC varied between 81-957. and 82-987. respectively, indicating that most DNA synthesis was mediated by DNA polymerase a, but that a small but significant amount of residual synthesis was indicated. A qualitative approach was then applied to determine whether the apparent residual DNA synthesis was restricted to a subpopulation of giant cells or whether all giant cells displayed a low level of DNA synthesis. Autoradiographs of the ICM of blastocysts and the embryo proper of 7.5da embryos, which acted as diploid control population, was completely inhibited regardless of duration in explant culture. In contrast, primary trophoblast giant cells derived from blastocysts and secondary giant cells derived from the EX and EPC were observed to possess some heavily labelled cells after aphidicolin treatment. These results suggest that although DNA polymerase a is the primary replicating enzyme responsible for endoreduplication in mouse trophoblast giant cells, some nonactivity is also observed. A DNA polymerase assay employing tissue lysates of outgrown 7.5da embryo, EX and EPC tissues was used to attempt to confirm the presence of higher nonactivity in tissues possessing trophoblast giant cells. Employing a series of inhibitors of DNA polymerases, it would appear that DNA polymerase a is the major polymerase active in all tissues of the 7.5da mouse embryo. The nature of the putative residual DNA synthetic activity could not be unequivically determined in this study. Therefore, these results suggest that both primary and secondary trophoblast giant cells possess and use DNA polymerase a in endoreduplicative DNA synthesis. It would appear that the high levels of DNA polymerase g activity reported in trophoblast tissue derived from the 13.5 da rat yolk sac was not a general feature of all endoreduplication.

The influence of physical activity behaviour on the relationship between motor proficiency and body composition in children

There is an emerging awareness that children with poor motor abilities are at particular risk for overweight. This cross-sectional study examined the influence of physical activity behaviour on the relationship between motor proficiency and body composition. Participants were 1287 (646 males, 641 females) Grade 6 students in the Physical Health Activity Study project. Height, weight, waist girth, and motor proficiency (Bruininks-Oseretsky Test of Motor Performance BOTMP-SF) were assessed. Physical activity behaviours were also evaluated with a multifaceted approach and reported for school-based, non-school based physical activity, free-time play, and sedentary activities (Participation Questionnaire), and leisure time exercise (Godin-Shephard Leisure Time Exercise Questionnaire GS). Overweight was defined by BMI scores: boys :::20.6-21.2 and <25.1-26.0; girls: ::: 20.7-21.7and <25.4-26.7 and obesity was defined as: boys:::: 25.1-26.0; girls: :::25.4-26.7. Children were classified as case group (CG,::; 10% on BOTMP-SF), borderline case group (BC, > 10% to ::; 20% on BOTMP-SF) or non-case group. Analyses of variance (ANOVAs) uncovered a significant difference in overweight and obesity between the case group and non-case group. Normal-weight children reported higher participation in organized school-sports (intra-mural and inter-school teams). The CG reported significantly lower participation in school sports teams and lower GS results, with a trend towards lower participation in all active pursuits. They also reported a significantly higher duration of television watching and book reading. There were no significant differences between motor proficiency groups by gender, age, nonschool sports, or free-time activity. Multivariate ordinal logistic regression analysis showed that the case group was 10.9 times more likely to be overweight/obese than their peers. No single aspect of physical activity was able to explain the difference in odds ratios for the motor proficiency groups. However, for the entire cohort, children who participated in more organized school sports were less likely to be overweight/obese. These findings confirm that children with low motor proficiency are at significant risk of developing overweight. It is evident that these children have generally attenuated activity levels and heightened levels of sedentary pursuits. School-based activities appear particularly limited, and are the one area where children have near autonomy in their decision to pursue active opportunities. The promotion of school-based programs, specifically intramural sports may be an important aspect in increasing children's overall activity levels. It is also essential to consider the needs of those children with low motor proficiency when designing activity promotion programs. Future research should further explore motor proficiency and overweight/obesity.

Patterns in space : a regional study of motif in Minoan wall painting

This thesis consists of a quantitative analysis of the regional prevalence of certain artistic motifs as they appear in Minoan wall painting of the Neopalatial period. This will help to establish the relative degree of artistic autonomy exercised by each of the sites included in this study. The results show that the argument for itinerant artists during this time period is a strong one, but the assumption that these travelling artists were being controlled by any one palace-centre is erroneous. Rather, the similarities and differences seen suggest that the choices were predicated either by the specific patrons, or by the function of the associated building or room. Thus, the motifs found within this study should be understood as constituting a cultural identity, with greater or lesser degrees of regional homogeneity, which act as one facet of a number of cultural indicators that can be used to better understand the role of artists and regional dynamics on the island during the Bronze Age.

Welland County Records, 1961-1989

The origins of the Welland County Fair date back to the founding of the County of Welland in 1852. A provincial charter was issued in 1853 to create the Welland County Agricultural Society that was to operate the Fair. In 1970, the Welland County Fair became the Niagara Regional Exhibition, and the Society became known as the Niagara Regional Agriculture Society. The Society seeks to “encourage interest, promote improvements in and advance the standards of agriculture, domestic industry and rural life”. The Welland Festival of Arts was developed in 1986 in order to revitalize the town’s economy. An “outdoor art gallery” was created by painting murals on buildings that depicted the town’s heritage, a concept successfully adopted by the town of Chemainus, B.C. The first mural was completed in the summer of 1988, and by 1991 there were a total of 28 murals around the city. The endeavour proved successful: in the years that followed the creation of the Festival, two new hotels were constructed, a third was expanded, and there was an addition to the Seaway Mall to accommodate the increased tourist traffic. Optimist International is a non-profit organization that strives to “bring out the best in kids” . The first Canadian club was formed in Toronto in 1924. The Welland branch of the Optimist Club was founded in 1937. The first Welland County General Hospital opened in 1908. As the population increased, it became necessary to expand the existing facilities. Additions were made to the original structure with an East wing in 1930 and a children’s ward in 1931. However, in the 1950’s, the hospital was operating beyond optimum capacity and the need for a larger facility was clear. It was decided that a new hospital would be built, which opened in April 1960. The new hospital had 259 beds and 51 bassinets. Further additions were made in 1967 and 1978. The County of Welland was formed in 1850 when it was officially separated from Lincoln County, however, the two counties continued to operate together until 1856 when a new County building and jail for Welland County were completed. That same year, the first meeting of the Council of the Corporation of Welland County took place. The final meeting of the Council took place on December 18, 1969. The following year, the County of Welland merged with Lincoln County to form the Regional Municipality of Niagara. The Welland Mills in Thorold, Ont., was built in 1846-1847 by Jacob Keefer and is thought to have been one of the largest flour mills in Upper Canada. Ownership of the mill changed several times over the years and previous owners include the Howland family, the Hedley Shaw Milling Company and the Maple Leaf Milling Company. In 1986, the building received a heritage plaque from the Ontario Heritage Foundation, an agency of the Ontario Ministry of Culture and Recreation. At this time, the mill was no longer in operation and was being used for storage by Fraser, Inc. By 2006, the dilapidated building had been redeveloped into18 apartments and 2 floors of commercial space, while maintaining many heritage features. The building is currently known as the Welland Mills Centre.

Influence de l’agrégation érythrocytaire sur la migration axiale de microparticules simulant des plaquettes sanguines

Lors du phénomène d’hémostase primaire ou de thrombose vasculaire, les plaquettes sanguines doivent adhérer aux parois afin de remplir leur fonction réparatrice ou pathologique. Pour ce faire, certains facteurs rhéologiques et hémodynamiques tels que l’hématocrite, le taux de cisaillement local et les contraintes de cisaillement pariétal, entrent en jeu afin d’exclure les plaquettes sanguines de l’écoulement principal et de les transporter vers le site endommagé ou enflammé. Cette exclusion pourrait aussi être influencée par l’agrégation de globules rouges qui est un phénomène naturel présent dans tout le système cardiovasculaire selon les conditions d’écoulement. La dérive de ces agrégats de globules rouges vers le centre des vaisseaux provoque la formation de réseaux d’agrégats dont la taille et la complexité varient en fonction de l’hématocrite et des conditions de cisaillement présentes. Il en résulte un écoulement bi-phasique avec un écoulement central composé d’agrégats de globules rouges avoisinés par une région moins dense en particules où l’on peut trouver des globules rouges singuliers, des petits rouleaux de globules rouges et une importante concentration en plaquettes et globules blancs. De ce fait, il est raisonnable de penser que plus la taille des agrégats qui occupent le centre du vaisseau augmente, plus il y aura de plaquettes expulsées vers les parois vasculaires. L'objectif du projet est de quantifier, in vitro, la migration des plaquettes sanguines en fonction du niveau d’agrégation érythrocytaire présent, en faisant varier l’hématocrite, le taux de cisaillement et en promouvant l’agrégation par l’ajout d’agents tels que le dextran à poids moléculaire élevé. Cependant, le comportement non Newtonien du sang dans un écoulement tubulaire peut être vu comme un facteur confondant à cause de son impact sur l’organisation spatiale des agrégats de globules rouges. De ce fait, les études ont été réalisées dans un appareil permettant de moduler, de façon homogène, la taille et la structure de ces agrégats et de quantifier ainsi leur effet sur la migration axiale des plaquettes. Du sang de porc anti coagulé a été ajusté à différents taux d’hématocrite et insérer dans un appareil à écoulement de Couette, à température ambiante. Les plaquettes sanguines, difficilement isolables in vitro sans en activer certains ligands membranaires, ont été remplacées par des fantômes en polystyrène ayant un revêtement de biotine. La quantification de la migration de ces fantômes de plaquettes a été réalisée grâce à l’utilisation de membranes biologiques fixées sur les parois internes de l’entrefer du rhéomètre de Couette. Ces membranes ont un revêtement de streptavidine assurant une très forte affinité d’adhésion avec les microparticules biotynilées. À 40% d’hématocrite, à un cisaillement de 2 s-1, 566 ± 53 microparticules ont été comptées pour un protocole préétabli avec du sang non agrégeant, comparativement à 1077 ± 229 pour du sang normal et 1568 ± 131 pour du sang hyper agrégeant. Les résultats obtenus suggèrent une nette participation de l’agrégation érythrocytaire sur le transport des fantômes de plaquettes puisque l’adhésion de ces derniers à la paroi du rhéomètre de Couette augmente de façon quasi exponentielle selon le niveau d’agrégation présent.

Les cycles d'azulejos de l'église du couvent de Louriçal (Portugal) attribués à Valentim de Almeida (1692-1779)

Pour respecter les droits d'auteur, la version électronique de ce mémoire a été dépouillée de ses documents visuels et audio-visuels. La version intégrale du mémoire a été déposée au Service de la gestion des documents et des archives de l'Université de Montréal

Fabrication des films microstructurés et leurs caractéristiques en spectroscopie de résonance des plasmons de surface

Cette thèse caractérise les propriétés optiques des matériaux plasmoniques microstructurés et procède à l’évaluation des paramètres analytiques afin de les employer comme plateforme de biodétection en spectroscopie de résonance des plasmons de surface (SPR). Aux dimensions micrométriques, les matériaux plasmoniques présentent des caractéristiques optiques propres aux nano- et macromatériaux. La cartographie physicooptiques en SPR de matériaux méso- et microscopiques s’est effectuée à l’aide de films structurés de motifs périodiques triangulaires et circulaires fabriqués par une technique modifiée de lithographie par nanosphères (nanosphere lithography, NSL). À partir de cette vue d’ensemble, quelques films structurés ont été sélectionné en fonction d’aspects analytiques tels que la sensibilité et la résolution face aux variations d’indice de réfraction (RI) pour déterminer le potentiel de ces matériaux comme plateforme de biodetection. Les propriétés optiques distinctes des films microstructurés proviennent d’interactions résonantes entre les modes de plasmons de surface (SP) localisé et délocalisé identifiés par la relation de dispersion en SPR ainsi que l’imagerie Raman. Les conditions de résonance des modes SP dépendant de paramètres expérimentaux (λ, θ, η) tel qu’observés numériquement par rigorous coupled wave analysis (RCWA) et empiriquement. Ces travaux démontrent la nature plasmonique distincte des micro-matériaux et leur potentiel d’intégration aux techniques analytiques SPR existantes. Les matériaux plasmoniques micrométriques furent également étudiés pour l’implémentation de la SPR à une pointe de microscopie à force atomique (atomic force microscopy, AFM) combinant ainsi la spectroscopie à l’imagerie topographique. Des travaux préliminaires se sont concentrés sur la signature spectroscopique de leviers en silicium (Si) et en nitrure de silicium (Si3N4), l’impact d’un revêtement d’or sur les pointes et l’influence de milieu environnant. Une image d’origine plasmonique a été obtenue avec des leviers en Si3N4 revêtus d’or en transmission dans un environnement aqueux, indiquant ainsi le potentiel de ces pointes comme micro-biocapteur SPR. Ces résultats préliminaires servent de fondement pour orienter les prochaines investigations dans ce projet.

Régulation de la fonction ovarienne par la voie de signalisation des WNTs

Les WNTs sont des glycoprotéines sécrétées impliquées dans plusieurs processus tels que la spécification cellulaire, la prolifération, la différenciation, et beaucoup d’autres. Pour transmettre leur signal, les WNTs se lient aux récepteurs Frizzled (FZD) et au co-récepteur « Low-density-lipoprotein receptor Related Protein » (LRP) 6 ou 5, activant ainsi l’une des trois principales voies de signalisation: la voie de signalisation WNT/β-caténine (voie canonique), la voie « Planar Cell Polarity » (PCP) et la voie WNT/Ca2+ (voies non-canoniques). Des antagonistes de cette voie, les « Secreted Frizzled Related Protein » (SFRPs), peuvent aussi se lier aux WNTs pour empêcher leur liaison aux récepteurs FZDs. Bien que des rôles importants aient été associés à plusieurs composants de la voie des WNTs lors de la régulation de l’ovaire adulte, le fonctionnement exact de cette signalisation reste nébuleux. L’objectif global de cette thèse visait donc à mieux comprendre la voie de signalisation des WNTs au niveau de l’ovaire adulte de souris, par la caractérisation de deux autres composants de cette voie, FZD1 et SFRP4. La création et l’analyse de souris Fzd1 et Sfrp4 KO ont démontré que FZD1 est nécessaire pour la régulation des gènes associés à l’expansion du cumulus, dans le complexe ovocyte-cumulus (COC). Nous avons aussi constaté que SFRP4 avait un rôle à jouer lors de la régulation des gènes associés à l’expansion du cumulus mais cette fois, au niveau des cellules de la granulosa. Finalement, les résultats in vivo et in vitro de cette étude ont suggéré que la voie PCP, contrairement à la voie canonique, pourrait être modulée dans les cellules de la granulosa des souris Sfrp4 KO, possiblement grâce au signal induit par WNT4 et WNT5a. Ces données ont permis de créer un modèle hypothétique représentant la régulation de la signalisation ovarienne par les WNTs. Ce modèle servira de base pour l’élaboration de futurs projets de recherche visant à comprendre davantage la signalisation ovarienne et les possibles effets de sa dérégulation lors de processus pathologiques. Ces connaissances pourront ensuite être appliquées chez l’humain afin de traiter plusieurs maladies ou dérèglements ovariens.