Poor skeletal health results from aging and metabolic diseases such as obesity and diabetes and involves impaired homeostatic balance between marrow osteogenesis and adipogenesis. Tissue engineering provides researchers with the ability to generate improved, highly controlled and tailorable in vitro model systems to better understand mechanisms of homeostasis, disease, and healing and regeneration. Model systems that allow assembly of modules of MSCs, osteoblasts, and adipocytes in a number of configurations to engage in signaling crosstalk offer the potential to study integrative physiological aspects and complex interactions in the face of changes in local and systemic microenvironments. Thus, the overall goal of this dissertation was to examine integrative physiological aspects between MSCs, osteoblasts, and adipocytes that exist within the marrow microenvironment. To investigate the effects of intercellular signaling in different microenvironmental contexts, methods were developed to photolithographically pattern and assemble cell-laden PEG-based hydrogels with high spatial fidelity and tissue-scale thickness for long-term 3D co-culture of multiple cell types. This platform was applied to study effects of crosstalk between MSCs, osteoblasts and adipocytes on markers of differentiation in each cell type. Additionally, responses of MSCs to systemic perturbations in glucose concentration were modulated by mono-, co-, and tri-culture with these cell types in a model of diabetes-induced skeletal disease. Together, these studies provided valuable insight into unique and differential effects of intercellular signaling within the niche environment of MSCs and their terminally differentiated progeny during homeostatic and pathological states, and offer opportunities further study of integrative physiological interactions between mesenchymal lineage cells.

Die vorliegende Arbeit beschäftigt sich mit der mechanischen Untersuchung des menschlichen Fersenfettpolsters (FP) in Abhängigkeit von Alter und Geschlecht. Das menschliche FP stellt evolutionsgeschichtlich eine Anpassung an den aufrechten Gang dar. Durch Aufrichtung des Oberkörpers im Zweibeingang kam es zur Ganglinienverlängerung in Richtung Ferse und folglich zu einer Mehrbelastung des Rückfußes. Sie prägten die Funktion des FP, die Kräfte beim Aufsetzen der Ferse zu reduzieren. Das FP, das ein spezielles Unterhautfettgewebe ist und sich aus straffem und lockerem Bindegewebe zusammensetzt, kann die Kräfte durch Verteilen und Absorbieren vermindern. Bekannt ist, dass sich das mechanische Verhalten der Haut zwischen Mann und Frau unterscheidet. Da das FP ein Bestandteil der Haut ist, stellt sich als ein Schwerpunkt dieser Arbeit die Frage: Unterscheidet sich das FP mechanisch zwischen Mann und Frau? Aus naturwissenschaftlicher Sicht ist Altern ein natürlicher Mechanismus, der die Lebenskraft des Organismus durch Zellalterung und –tod reduziert. Aus evolutionärer Sicht wurde zugelassen, dass sich bestimmte Zellen bzw. Gewebe, welche hohen endogenen und exogenen Faktoren ausgesetzt sind, vollständig aber begrenzt regenerieren dürfen. Es wird als primäres Altern gekennzeichnet. Das primäre Altern kann positiv oder negativ durch äußere Einwirkungen auf den Organismus („Sekundäres Altern“) beeinflusst werden. Bindegewebe, welches hohen exogenen Faktoren ausgesetzt ist, sind insbesondere Schnittstellen zwischen „Biologischem System“ und „Umwelt“ (z.B. beim Menschen das Gewebe der Hautinnenfläche oder der Fußsohle). Es wird erwartet, dass das FP dem Alterungsprozess stark unterworfen ist, da es alltäglich mechanisch beansprucht wird. Folglich kann es zu einem mechanischen Funktionsverlust des FP kommen, das sich negativ auf die Belastbarkeit des Rückfußes auswirken kann. Die Entwicklung von altersbedingten Verschleißerkrankungen kann nicht ausgeschlossen werden. Als ein zweiter Schwerpunkt dieser Arbeit stellt sich die Frage: Unterscheidet sich das FP mechanisch zwischen Jung und Alt? Es gibt hinsichtlich der Thematik dieser Dissertation wenige Untersuchungen. Die Autoren kommen zum Teil zu unterschiedlichen Ergebnissen und Schlussfolgerungen, dass möglicherweise auf die unterschiedlich verwendete Methodik zurückzuführen ist. Die Vor- und Nachteile der bisher durchgeführten Experimente machen es schwierig, Stellungnahme zu beziehen, welche der Tests die zuverlässigsten Ergebnisse liefern. Seit den letzten 10-15 Jahren wurde immer häufiger Ultraschall als zusätzliche Informationsquelle in mechanischen Messplätzen integriert, um innere Kenndaten zum mechanischen Verhalten des FP abzuleiten. Allerdings waren die quasi-statischen Messungen und die geringen Kontaktkräfte der limitierende Faktor um das mechanische Verhalten valide zu charakterisieren. Mit einem eigens entwickelten Messplatz sollte dieser methodische Ansatz überholt werden. Der instrumentierte Belastungsschlitten ermöglicht die Aufnahme von dynamischen Ultraschallbildsequenzen unter mindestens 10-fach höheren Kontaktkräften bei fast doppelter Fersenkontaktgeschwindigkeit gegenüber den bisher bekannten Ultraschallexperimenten in der Literatur. Mögliche geschlechts- und altersspezifische Unterschiede im mechanischen Verhalten des FP sind grundlegend für die Orthopädie-Technik, die klinische Forschung und die Biogerontologie. Die Orthopädie-Technik benötigt insbesondere die Erkenntnisse zum mechanischen Verhalten der Haut an unterschiedlichen Stellen der unteren Extremität in Abhängigkeit von Alter und Geschlecht, um den Tragekomfort und die Bewegungseffizienz von Prothesen und Orthesen zu bessern. In der klinischen Forschung zeigt sich das Interesse an den altersspezifischen mechanischen Kenndaten, um im Zusammenhang zu klinischen Parametern die Entwicklung von orthopädischen Erkrankungen zu erforschen. Für die Biogerontologie wäre diese Art von Forschung relevant, um Zusammenhänge zu histologischen Parametern zu überprüfen, die direkt am Alterungsprozess des Bindegewebes beteiligt sind. Sie könnten zur Entschlüsselung des Mechanismus „Altern“ beitragen.

近幾年的大規模社會抗爭運動，不像過往傳統社運組織由上而下的動員，強調一致的意識形態和認同作為號召，而是透過網際網路與社交媒體的訊息擴散與中介動員，形成水平式的串連。面對這種新型態的網絡化社會運動，Bennett & Segerberg（2013）提出連結性行動（connective action）概念，強調個人化行動框架的重要性，以及組織在這些新型態社會運動中扮演的不同角色。本研究以2013年苗栗大埔農地徵收一案作為本地的連結性行動個案，觀察社交媒體上的浮現社群和既有的社運組織之連結，探討線上與線下參與者之串連如何使此案成為眾所關注的社會抗爭事件。 　　本研究採取複合式研究方法進行資料蒐集與分析，研究場域包括線上社交媒體與線下田野觀察，並訪談實際參與者。研究發現，在2013年苗栗大埔事件中，透過社交媒體的訊息擴散與動員參與，一波又一波的訊息擴散累積為下次的動員能量，號召更多的個別行動者參與；再者，社運組織作為一個連結平台，讓線上社交媒體的議題社群，有機會與在地抗爭者共同參與活動，並捲動不同社運組織之間的連結，形成社運組織「網絡的網絡」的協作圈。 / In the 2013, residents of Miao-li Da-pu fought for their living right against the government. They organized a series of protest by mobilizing people through social media. This was one of the several high-profile networked social movements in recent years. Adopting “the logic of connective action” from Bennett & Segerberg (2013) as the conceptual framework, this study examines the online and offline social networks of activist organizations during the protest events of Miao-li Da-pu. This analysis indicates that the protest’s information flows on the social media may recruit more individuals to participate the protest. Moreover, the organizations enabled “the network of networks”, which is linked to the networks belonging to different organizations. The role of organization in the protest is not only to mobilize the resources but also an enrolling platform to connect various individuals and local residents to work together.

La prolongation de l’intervalle électrocardiographique QT est un facteur de risque d’arythmie ventriculaire et de mort subite. Cette anomalie, retrouvée chez certains patients atteints de lupus érythémateux disséminé, pourrait contribuer à la mortalité cardiovasculaire élevée dans cette population. L’anti-Ro/SSA, un auto-anticorps retrouvé chez environ 30% des patients atteints de lupus, est associé à la présence de blocs cardiaques chez le nouveau-né et pourrait aussi augmenter le risque de prolongation pathologique de l’intervalle QT chez l’adulte. Le présent mémoire est constitué de cinq chapitres traitant de l’association potentielle entre l’anticorps anti-Ro/SSA et la prolongation de l’intervalle QT. Le premier chapitre constitue une introduction permettant de mettre en contexte les éléments essentiels à la compréhension du projet d’étude. Le deuxième chapitre constitue une revue de l’état des connaissances actuelles sur le lien potentiel entre anti-Ro/SSA et intervalle QT. Le troisième chapitre présente le projet d’étude par l’intermédiaire d’un article publié dans Arthritis Care and Research. Dans cette étude, les patients de la cohorte de lupiques du Centre Universitaire de santé McGill ont subi des électrocardiogrammes dans l’objectif d’estimer l’association entre l’anti-Ro/SSA et les anomalies électrocardiographiques, en tenant compte d’autres facteurs démographiques et cliniques. L’association entre la prolongation de l’intervalle QT et la présence de l’anti-Ro/SSA a été démontrée (rapports de cotes ajustés de 5.1 à 12.6) et les patients porteurs de l’anti-Ro/SSA pourraient donc bénéficier de dépistage électrocardiographique systématique. Les points faibles et forts de cet article sont discutés dans le quatrième chapitre et des perspectives de recherches futures sont finalement abordées. / QT interval prolongation on the electrocardiogram is a risk factor for ventricular arrhythmias and sudden cardiac death. This abnormality is found in patients with systemic lupus erythematosus and could contribute to the high cardiovascular mortality rate in this population. Anti-Ro/SSA is an auto-antibody presents in about 30% of lupus patients and is associated with congenital cardiac block. This auto-antibody could also increase the risk of pathologic prolongation of the QT interval in adults. This master’s thesis is comprised of five chapters discussing the potential association between anti-Ro/SSA antibody and QT interval prolongation. The first chapter is an introduction to the essential elements for the understanding of the study project. The second chapter is a literature review of the potential link between anti-Ro/SSA and QT interval prolongation. The third chapter presents the study project through an article published in Arthritis Care and Research. In this study, patients from the McGill lupus cohort were invited to undergo electrocardiograms in order to estimate the association between anti-Ro/SSA antibody and electrocardiographic abnormalities, while taking into account the other potentially associated demographic and clinical factors. This study shows an association between anti-Ro/SSA and prolonged QT interval (Odds ratios: 5.1 to 12.6) and patients positive for anti-Ro/SSA may thus benefit from electrocardiographic testing. Strengths and weaknesses of this article are discussed in the fourth chapter and future research areas are finally explored.

No description available.

Prior immunity to a related or unrelated pathogen greatly influences the host’s immune response to a subsequent infection and can cause a dramatic difference in disease course, a phenomenon known as heterologous immunity. Heterologous immunity can influence protective immunity, immunopathology and/or immune deviation of cytokine-producing T cell subsets. Examples of heterologous immunity have been well documented in mouse models, as well as during human infections. For example, prior immunity to lymphocytic choriomeningitis virus (LCMV) provides partial protection against vaccinia virus (VV), as LCMV-immune mice show reduced VV titers and increased survival upon lethal dose VV infection. Heterologous protection against VV challenge, as a result of LCMV immunity, is mediated by LCMV-specific CD4 and CD8 T cells, as transfer of LCMV-specific memory T cells can mediate this protective effect in naïve mice. The recognition of a single TCR with more than one MHC-peptide complex is referred to as T cell cross-reactivity. A VV Kb-restricted epitope a11r198 was identified to be able to induce cross-reactive responses from LCMV-specific CD8 T cells. During VV infection, LCMV-specific memory T cells that are cross-reactive to VV epitopes produce IFN-γ early in VV infection. IFN-γ is essential for mediating the protection against VV in LCMV-immune mice, as this heterologous protection is absent in IFN-γR-/-and IFN-γ blocking antibody-treated LCMV-immune mice. In addition to protective immunity, cross-reactive LCMV-specific memory T cells and IFN-γ also induce an altered immunopathology during heterologous VV challenge. LCMV-immune mice show moderate to severe levels of inflammation of the fat tissue, known as panniculitis, in the visceral fat pads upon VV challenge. In humans, panniculitis is a painful condition, most commonly presenting as erythema nodosum. Erythema nodosum is a disease of unknown etiology with no known treatment. It may occur following intracellular bacterial and viral infections, and occasionally happens after vaccination with VV for smallpox. During infections there can be a delicate balance between the ability of immune responses to provide protective immunity, and the tendency to induce immunopathology. By using the mouse model of heterologous immunity between LCMV and VV, we tried to understand how the immunity to LCMV biased the balance between the protective immunity and immunopathology, and what effector molecules were responsible for the pathogenesis of panniculitis in this system. TNF is a pleiotropic cytokine, which is required for normal innate and adaptive immune responses. Its functions range from inducing proliferative responses including cell survival, to destructive responses such as promoting apoptosis and programmed necrosis. In response to inflammatory stimuli, activated macrophages/ monocytes produce large amounts of TNF, and upon activation, T cells, B cells and NK cells also produce TNF. In vitro and in vivo studies have shown that TNF in synergy with IFN-γ plays an important role in mediating host defense against pathogens, such as Listeria monocytogenesand poxviruses in mice and hepatitis B virus and human immunodeficiency virus in humans. However, inappropriate expression of TNF often results in tissue damage. Considering the important role TNF plays in both host defense and mediating autoimmune diseases, we hypothesized that TNF was required for mediating both protective and pathogenic effects in the heterologous immunity between LCMV and VV. We first examined whether TNF was involved in mediating protective heterologous immunity. LCMV-immune mice, that were TNF-deficient as a consequence of genetic deletion (TNF-/-) or receptor blockade by treatment with etanercept (TNFR2: Fc fusion protein), were challenged with VV. These TNF-deficient mice showed normal recruitment and selective expansion of cross-reactive LCMV-specific memory CD8 T cells. They also exhibited efficient clearance of VV similar to LCMV-immune mice with normal TNF function. Thus, we concluded that neither TNF nor lymphotoxin (LT), which uses the same receptors as TNF, was required in mediating protective heterologous immunity against VV. Indeed, prior immunity to LCMV could completely compensate for the role of TNF in protection of naïve mice against VV infection, even under conditions of lethal dose inoculum. Thus, heterologous immunity may help explain why treatment of humans with etanercept is reasonably well tolerated with relatively few infectious complications. One of the histological characteristics of panniculitis is necrosis of adipose tissue. It is known that three members in the TNF superfamily, i.e. TNF/LT, FasL and TRAIL are able to induce necrosis of a target cell. It is also known that TNF is able to induce VV-infected cells to go through necrosis, when apoptosis is blocked in these cells by VV protein. Furthermore, TNF and FasL have already been shown to be associated with some skin and fat pathology. Thus, we hypothesized that TNF, FasL and TRAIL were involved in the pathogenesis of panniculitis in VV infected LCMV-immune mice. By using blocking antibodies or genetically deficient mice, we demonstrated that both TNF/LT and FasL were crucial for inducing panniculitis. Although TNFR1 has been reported to induce programmed necrosis, our data indicated that TNFR2, not TNFR1, was involved in mediating tissue damage in the fat pads of LCMV-immune mice infected with VV. We also found that TNF signaled through TNFR2 to up-regulate the expression of Fas on adipocytes. Thus, the engagement of Fas on the adipocytes with FasL expressed on activated VV-specific and cross-reactive LCMV-specific CD8 T cells in the fat pads could lead to panniculitis. Thus, our data may identify a potential mechanism in the pathogenesis of human panniculitis, and may suggest a possible treatment for this painful disease. Recent reports suggest that heterologous immunity may contribute to the tremendous variation in symptoms between individuals, from subclinical to death, upon viral infection. Even in genetically identical mice, variations in immunopathology from none to life-threatening levels of pathology are observed in LCMV-immune mice during VV infection. By adoptive transfer of splenocytes from a single LCMV-immune donor into two recipients, we showed that similar levels of pathology were generated in mice receiving the same splenocytes. However, the level of pathology varied among recipients receiving splenocytes from different LCMV-immune donors. The difference in levels of VV-induced pathology observed in individual LCMV-immune mice was a reflection of the private specificity of the T cell repertoire, which is a unique characteristic of each individual immune host. The goal of this doctoral thesis is to understand how heterologous immunity contributes to the pathogenesis of panniculitis. Our data demonstrate that TNF/LT and FasL directly contribute to development of panniculitis in LCMV-immune mice during VV infection, and suggest that anti-TNF treatment might be a useful treatment for diseases, such as erythema nodosum and lupus-induced acute fatty necrosis in humans.

Osteogenesis imperfecta, innate brittle bone disease, is a very serious disease. It is inheritable disease of connective tissue, which shows by abnormal fragility of bones. The occurrence of this disease is one case in 10 000 30 000 births. The theoretical part of the thesis deals with the disease itself, also the psychical impact on children suffering from Osteogenesis imperfecta and the impact on their families as well. At the beginning of the research, three goals of this thesis were set: map out (on the basis of theoretical and practical backgrounds) the pitfalls of life of children with the disease Osteogenesis imperfecta, find out what are the most common difficulties by children with the disease Osteogenesis imperfecta and also find out the experiences of nurses with the care for children with disease Osteogenesis imperfecta. The empirical part of the thesis was processed by means of qualitative research conducted by the technique of semi-structured interview and narrative biographical interview. The research set were nurses working at the child departments in hospitals, parents of ill children and also an adult woman with the diagnosis of Osteogenesis imperfecta and two doctors. From the research emerged that among the most common difficulties of children is pain, which decreases the quality of their life. Small children can't engage in typical activities of children, such as going to a playground, older children can't attend for example music festivals. Children feel fear from fractures and are therefore limited in sports. Because of injuries and their treatments, the children have more absences at schools and therefore are isolated from peers. Nevertheless, the children with this disease can live a happy life. From the results of the research also emerges, that nurses working at the child departments of the hospitals attended by children with this illness have a good experiences with their treatment. They are able to give parents important information and know the specifics of application of the treatment. The results of the diploma thesis were presented at a national student conference and will be further published.

The aim of this diploma thesis was to summarize the topic of the anterior cruciate ligament (LCA) injury dependency on the menstrual cycle phase of young women with regular sport activity. The general part describes basic knowledge about connective tissue, LCA, issues of LCA injury and related risk situations. Large chapter is dedicated to sex hormones and menstrual and ovarian cycle, which is crucial for understanding the whole topic. The main part is focused on impact of sex hormones and hormonal contraception on connective tissue, but also on muscle and nervous tissues, which might be as well important for LCA injury incidence. Next part of the thesis consists of a questionnaire survey. 52 respondents aged 15-35 with rupture or partial rupture of LCA answered the non-standardized questionnaire compiled specially for this thesis and the results were statistically processed. 14 respondents were using hormonal contraception, remaining 38 had physiological menstrual cycle. Based on the theoretical findings we expected highest incidence of LCA injuries among women without contraception in phases of menstrual cycle with highest levels of oestrogen (10th -15th day). That was confirmed (P-value: 0,0218) as well as overall lower incidence among women using contraception (P-value: 0,0006). Expected higher...

Les modèles numériques personnalisables d'organes du corps humain offrent un formidable potentiel pour évaluer le risque lésionnel dans les domaines de la sécurité des transports, du médical ou du sport. Suivant les applications, différents niveaux de détails peuvent être nécessaires. En particulier, lorsque le comportement mécanique des tissus biologiques doit être finement reproduit, les modèles de comportement doivent intégrer des considérations sur la structure du tissu, et simuler les mécanismes suivant lesquels il réagit à un chargement mécanique. Le travail de thèse présenté ici s'est focalisé sur la capsule de foie, notamment sur ses propriétés microstructurales et mécaniques, afin d'identifier les hypothèses importantes à intégrer dans la construction d'un modèle constitutif de tissu fibreux basé sur la microstructure. La méthodologie expérimentale a été mise en place afin de caractériser le comportement mécanique de ce tissu, en lien avec l'organisation de sa microstructure. Des essais de traction uniaxiale et de gonflement sous microscope confocal biphotonique ont été développés, pour observer l'évolution de la microstructure sous chargement. Des déformations macroscopiques ont été mesurées, et une méthode de mesure de champs de déformations locaux a été développée pour quantifier l'état de déformation du réseau de fibres. La réorganisation du réseau de fibre de collagène a également été quantifiée. L'analyse des liens existant entre les grandeurs mesurées à l'échelle macroscopique et ces phénomènes microscopiques est proposée, pour préciser les hypothèses à adopter dans les modèles permettant de passer de l'échelle des fibres au comportement global du tissu / Customized human body models offer a great potential to assess the injury risks in the fields of transport safety, surgery or sport. Various detail levels can then be needed, according to the targeted application. In particular, when the mechanical behavior of biological tissues needs to be accurately reproduced, numerical models have to include information about the structure of the tissue, and model the mechanisms of the response to mechanical loading. The work presented here focuses on the microstructural and mechanical characterization of the human liver capsule, in order to identify the important hypotheses that need to be included in a fibrous tissue constitutive model, based on microstructure. Thus, an experimental methodology has been developed to identify the mechanical behavior of this particular tissue, related with its microstructural organization. Uniaxial tensile tests, as well as bulge tests under a multiphoton confocal microscope have been performed, to observe the microstructure evolution during loading. Macroscopic strain has been assessed, and a method to measure local strain fields has been developed, to quantify the strain state of the fibrous network. The reorganization of the collagen fibers network has also been quantified. An analysis of the links between the measured macroscopic parameters and the microscopic phenomena is given. Therefore, the hypotheses that need to be included in constitutive models are highlighted, with particular consideration given to the affine transformation hypothesis which allows to link the fibers behavior to the global response of the tissue

Chronic posterior compartment syndrome (CPCS) of the leg is a pathological condition which is often encountered by participants in exercise related activities such as running. To date no successful conservative treatment approach existed for the condition. The mainstay of the management of the condition at present is the surgical release of the involved fascia that surrounds the compartment. The main aim of the research project was thus to develop a successful conservative treatment approach for the symptoms of CPCS. It was identified that the current theoretical base did not incorporate the continuous and relatively inelastic nature of the fascia which plays an important role in the condition. Based on an extended literature review, muscles which are linked to the posterior compartment via the myofascial tissue were identified. Tightness in these clinically significant muscles is able to induce stresses in the myofascial chain which could ultimately influence stresses in the posterior compartment of the leg. The release of tightness in these muscles external to the posterior compartment through soft tissue mobilization techniques provides an effective conservative treatment approach for the symptoms of CPCS. A revised model for the pathogenesis of CPCS was developed which formed the basis for treatment interventions. The revised theoretical model for the pathogenesis of CPCS was validated based on a mixed-methodological approach which included a series of exploratory as well as explanatory case studies. This qualitative approach was supplemented by quantitative experiments in which the causal relationships of the condition on certain biomechanical aspects were explored. The treatment interventions had a hundred percent success rate and the results of the experimental research conducted also supports the new theoretical model for the pathogenesis of CPCS. / Thesis (DPhil)--University of Pretoria, 2008. / Biokinetics, Sport and Leisure Sciences / unrestricted