No sistema cardiovascular, modificações de pressão e shear stress devido ao fluxo sanguíneo influenciam a morfologia e a patofisiologia dos vasos sanguíneos e do coração. Neste trabalho, estudamos o papel de duas moléculas transmembrânicas do Sistema Renina-Angiotensina: a Enzima Conversora de Angiotensina (ECA) e o Receptor de Angiotensina do tipo I (AT1) como mecanosensoras e mecanotransdutoras dessas forças físicas. A ECA foi por muito tempo conhecida somente por sua ação em converter Angiotensina I em Angiotensina II e por inativar a Bradicinina. Recentemente foi demonstrado que a ECA, além dos efeitos enzimáticos já conhecidos, pode ter sua cauda citoplasmática fosforilada e desencadear vias de sinalização intracelular. Observamos que o shear stress, mas não o estiramento, induziu a diminuição da fosforilação da porção citoplasmática da ECA após 5 minutos de estímulo e se mantém até 18 horas. Demonstramos também que a porção extracelular da ECA tem papel fundamental como mecanosensora e que a via intracelular da JNK participa da mecanotransdução em resposta ao shear stress. Além disto, demonstramos que a diminuição da fosforilação da ECA está associada na diminuição da sua expressão pelo shear stress. O receptor AT1 é a principal molécula efetora das ações da angiotensina II. Recentemente foi demonstrado que esse receptor pode também ser ativado por forças físicas, estiramento celular, independentemente da presença da angiotensina II. No presente estudo, observamos que o receptor AT1 é ativado pelo shear stress e que o Candesartan, mas não o Losartan, é capaz de impedir esta resposta. A via intracelular ativada é dependente de proteína-G e da entrada de cálcio do meio extracelular. Interessantemente, a pré-exposicao dos receptores ao shear stress diminuem a responsividade dos receptores ao peptídeo Angiotensina II porém a Angiotensina II não é capaz de inibir a ativação pelo shear stress.. Em conjunto, demonstramos novos mecanismos de ação da ECA e do AT1 que são duas importantes moléculas do sistema renina angiotensina. A modulação destes componentes por estímulos mecânicos traz novas possibilidades de intervenções farmacologicas sobre esse sistema bem como o melhor entendimento da participação dessas moléculas na fisiopatologia cardiovascular. / Hemodynamic forces such as pressure and shear stress modulate the patophysiolgy of the cardiovascular system. In this study, we investigated two transmembranic key molecules of the renin-angiotensin system (RAS) as mechanosensors and mechanotransducers of physical forces: Angiotensin Converting Enzyme (ACE) and Angiotensin II type 1 Receptor (AT1). ACE is an enzyme that converts angiotensin I in angiotensin II. Recently, it was demonstrated that ACE cytoplasmic tail can be phosphorylated by ACE inhibitors and elicited intracellular cell signaling. Here, we observed that shear stress, but not stretch, decreased ACE cytoplasmic phosphorylation after 5 minutes and maintained up to 18 hours. ACE extracellular portion act as mechanosensor and JNK pathway participate in the mechanotransduction activation. In addition, we also demonstrate that decrease in ACE phosphorylation is involved in ACE expression downregulation by shear stress. AT1 receptor is the main effector molecule of angiotensin II cellular responses. It has recently been shown that AT1 receptor can directly be activated by mechanical stretch stress through an angiotensin-II-independent mechanism. In the present study, we observed that shear stress also activates AT1 receptor which is blocked by Candesartan, but not by Losartan. The intracellular pathway activated by shear stress involves both G-protein and extracellular calcium. Interestingly, preconditioning of AT1 receptor by shear stress impairs its responsiveness to angiotensin II while the pretreatment with angiotensin II still allow AT1 responsiveness to shear stress. Altogether, we demonstrated that ACE and AT1 receptor activates intracellular signal in response to mechanical force. This new concept for the RAS, the modulation of these molecules by mechanical forces gives new insigh into the discovery for pharmacological interventions to the RAS

Im 1997 wurden der Orphan GPR30 aus HUVECs kloniert, die FSS ausgesetzt waren. In dieser Studie konnte gezeigt werden dass die Expression von GPR30 durch die FSS-Behandlung im Vergleich zu unbehandelten HUVEC-Zellen deutlich induziert wurde. Daraufhin wurde in einer Studie von Isensse et al. die zelluläre und gewebsspezifische Expression von GPR30 in GPR30-LacZ Reportergen-Mäusen untersucht. Es konnte eine Expression von GPR30 vorwiegend in den Endothelzellen der kleinen Arterien verschiedenster Gewebetypen nachgewiesen werden. GPR30 war postuliert dass E2 direkt binden kann und dadurch rasche nicht-genomische Signale vermittelt. Im Gegensatz dazu haben verschiedene andere Veröffentlichungen gezeigt dass E2 nicht spezifisch an GPR30 bindet. Trotz der Kontroverse ob es sich bei GPR30 um einen Östrogenrezeptor oder nicht ist bislang nichts über seine Interaktion zu anderen Proteinen und deren Wechselwirkung bekannt. Deswegen war ein Ziel dieser Arbeit, Interaktionspartner von menschlichen GPR30 zu identifizieren und folglich ein humanes vaskuläres in vitro Modell zu etabliren, um die potentiellen Interaktionen von GPR30 sowie die downstream-Effekte der Wechselwirkung zwischen GPR30 und den neuen Interaktionspartner des vaskulären Modells auf Transkriptebene zu evaluieren. Ein Screening einer humanen kardiovaskulären cDNA-Bibliothek mit Hilfe des Y2H-Systems führte zur Identifizierung mehrerer Interaktionspartner für GPR30 darunter PATJ und FUNDC2. Durch anschließende CoIP konnte die Interaktion von GPR30 mit PATJ validiert werden. Des Weiteren konnte in dieser Arbeit die Wirkung von FSS auf die Expression von GPR30 in HUVECs bestätigt und ebenfalls in weiteren anderen Endothelzellen gezeigt werden. Abschließend wurde die Rolle von GPR30 und PATJ bei der Reaktion auf FSS auf transkriptioneller Ebene in HMEC-1-Zellen genomweit untersucht. Interessanterweise war eine Gruppe von Genen aufgrund von FSS in Zellen die GPR30 überexprimierten dereguliert als alleine durch FSS. / In 1997, the orphan G protein-coupled receptor 30, GPR30, was cloned using HUVECs exposed to FSS. It was shown that the level of GPR30 expression was up-regulated in response to FSS. Subsequently, in a study performed in the laboratory where the work for this thesis was carried out, the cellular and tissue distribution of GPR30 were investigated in GPR30-LacZ reporter mice and the expression was found predominantly in the endothelial cells of small arteries in several tissue types. GPR30, was also claimed to bind 17-β-estradiol (E2) directly and to mediate rapid non-genomic signalling. In contrast, various reports have indicated that E2 fails to bind GPR30 in a specific manner. Despite the controversy on whether GPR30 is an estrogen receptor or not, nothing is known at present about its relation and interaction with other proteins. Therefore, the aim of the work described in this thesis was to identify human GPR30 protein interaction partners and to establish a human vascular in vitro model in order to evaluate the potential role of GPR30 and the downstream effects of the interaction between GPR30 and new interaction partners in a vascular model at transcript level. The screening of a human heart cDNA library using the yeast two-hybrid assay led to the identification of several interaction partners for GPR30, among them PATJ and FUNDC2. These interactions were verified by CoIP experiments and the interaction of GPR30 with PATJ could be confirmed. The effect of FSS on the expression of GPR30 was confirmed in HUVECs and was detected in other endothelial cell types. In HUAECs, HAoECs and HMEC-1 cells GPR30 was also found up-regulated upon FSS, suggesting that GPR30 may indeed play a key role in vascular physiology. Finally, the role of GPR30 and PATJ in the FSS response was investigated at the genome-wide transcript level in HMEC-1 cells. Interestingly, a different panel of genes was deregulated owing to FSS in cells over-expressing GPR30 compared to FSS alone.

Altered flow regimes following river regulation can result in significant changes in river bed geomorphology and subsequent negative ecological impacts caused by re-suspended sediments deposited on the riverbed. This study aimed to evaluate the consequences of implementing an ecological flow regime on sediments accumulated within the regulated river Juktån. Sediments were sampled and analysed for particle size distribution to estimate sediment stability. Flow alteration following the ecological flow regime was analysed with HEC-RAS unsteady flow simulation serving as a basis for calculations of forces acting to erode or retain deposited sediments. Additional analyses regarding critical flow were made with HEC-RAS steady flow simulation. Results show that 4 out of 15 cross-sections analysed would have the potential to erode and re-suspend sediments. The estimated average critical flow for when sediments become unstable with potential to re-suspend is 17 m3/s. The total sediment inventory of the studied reach is ~25000 ton, with ~3000-ton sediments potentially eroding into re-suspension. This is approximately 3% of river Umeälvens annual 100 000 ton suspended sediments before being regulated. Results indicate that river bed heterogeneity in river Juktån could benefit from implementing the ecological flow regime while not mobilizing such amounts of fine sediments that would cause clogging effects downstream the site of interest. The study also introduces the erosion rate equation which compares the annual erosion between two different flow regimes.

L’hypertension artérielle (HTA) est l’une des pathologies les plus fréquentes et les plus préoccupantes des pays occidentaux. Elle est souvent associée au surpoids, à des maladies rénales, cardiaques et aussi à un dysfonctionnement du système endocrinien. Les désordres vasculaires compliquant l’HTA induite par le traitement par l’érythropoïétine recombinante (r-HuEPO) chez les patients ayant une maladie rénale chronique ou en lien avec une perturbation du système rénine-angiotensine-aldostérone (SRAA), associent une augmentation de la rigidité artérielle, une dysfonction endothéliale, un déséquilibre endothéline-1/monoxyde d’azote (ET-1/NO) et un état inflammatoire. L'inflammation vasculaire contribue à la physiopathologie de l'HTA par l’augmentation du stress oxydatif et l'activation des cellules immunitaires. Plusieurs études ont suggéré que le système immunitaire est impliqué dans le développement des maladies cardiovasculaires. Cependant le rôle des lymphocytes T régulatrices (Treg) dans l'HTA ou d'autres formes de maladies cardio-vasculaires, reste encore largement inconnu. Bien que la majorité des études ont montré que l’HTA induite par l’r-HuEPO est associée à une dysfunction endothéliale et à un déséquilibre du rapport (ET-1/NO), les mécanismes exacts restent à être identifier.Plusieurs études ont montré que l’exercice physique d’endurance prévient l’HTA chez les patients ou les modèles animaux. L’objectif de la première partie du travail est d’étudier l’impact du shear stress et de l’exercice sur les désordres vasculaires et l’HTA induits par l’r-HuEPO, en présence d’un déséquilibre endothélial (ET-1/NO). Pour ce faire, trois environnements ont été utilisés (in vitro (cellules en culture), in vitro (artères mésentériques (AM)) et in vivo (souris transgéniques sur-exprimant l’ET-1 au niveau de l’endothélium). Nos résultats montrent que l'association L-NAME/r-HuEPO est responsable d'une vasoconstriction flux-dépendante et une augmentation accrue du shear stress correspondant (plus de 25 dyn/cm2). Le Bosentan, inhibiteur non sélectif des récepteurs de l’ET-1, empêche la vasoconstriction flux-dépendante engendrée par la combinaison L-NAME/rHu-EPO, sans pour autant corriger la vasodilatation; Le traitement des souris transgéniques eET-1 avec l’r-HuEPO augmente la pression artérielle systolique, la concentration plasmatique en ET-1, le stress oxydatif, l’infiltration des monocytes et des macrophages aortiques (MOMA-2), le taux des cytokines pro-inflammatoire INF-γ, TNF-α et IL-6 et exacerbe la dysfonction endothéliale. L’exercice physique prévient tous les effets délétères engendrés par l’administration de l’r-HuEPO. Il est à noter aussi que l’exercice augmente le taux du Foxp3 dans la rate et le cortex rénal. Quant à la deuxième partie, il s'agissait de tester l’effet du transfert adoptif des Treg sur l’HTA et les désordres vasculaires induits par l’administration d’angiotensine II ou d’aldostérone à des souris. Nos résultats montrent que le transfert adoptif des Treg prévient l’HTA induite par l’Ang II, l’altération de la vasodilatation endothélium-dépendante, prévient l’augmentation de la rigidité des AM, diminue le stress oxydatif et les taux plasmatiques des cytokines proinflammatoires (INF-γ, TNF-α et IL-6), ainsi que l’infiltration aortique et rénale des macrophages. Le transfert adoptif des Treg prévient partiellement l’augmentation de la pression artérielle systolique induite par l’Aldo, prévient l’altération de la vasodilatation endothelium dépendante et le remodelage hypertrophique des AM, diminue le stress oxydatif et l’infiltration des cellules immunitaires inflammatoires. Le transfert adoptif des cellules T effectrices (Teff) exacerbe la majorité des effets de l’Aldo. Nos résultats justifient, d’une part, l’importance de l’exercice comme outil préventif de l’HTA induite par l’r-HuEPO, et d’autre part, présentent les Treg comme élément essential dans la modulation de l’HTA et des désordres cardiovasculaires. / Hypertension (HTN) is one of the most frequent disease and is of greatest concern in Western countries.HTN is often associated with overweight, kidney, heart and endocrine system disease. Vasculardisorders, associated with HTN induced by erythropoietin (r-HuEPO) treatment in chronic kidneydisease or in case of disruption of the renin-angiotensin-aldosterone system (RAAS), associated increasein arterial stiffness, endothelial dysfunction, unbalanced endothelin-1/nitric oxide ratio (ET-1/NO) andinflammation. Vascular inflammation contributes to the pathophysiology of hypertension by increasingoxidative stress and activation of immune cells. Several studies have suggested that the immune systemis involved in the development of cardiovascular disease. However, the role of regulatory T cells (Treg)in HTN and other forms of cardiovascular diseases remains largely unknown. In addition, most of thestudies have shown that hypertension induced by r-HuEPO is related to endothelial dysfunction and theratio (ET-1/NO). The exact mechanisms remain to be identified. Several studies have shown thatphysical exercise prevents HTN in patients or animal models. The aim of the first part of this work is tostudy the impact of shear stress and exercise on HTN and vascular disorders induced by r-HuEPO, in thepresence of an endothelial imbalance (ET-1/NO). This was carried out in three settings: in vitro (culturecells), ex vivo (mesenteric arteries) and in vivo (transgenic mice overexpressing ET-1 in endothelialcells). Our results show that the association L-NAME/r-HuEPO is responsible for a significant decreasein intravascular diameter, in response to elevated intravascular flow resulting in a flow-dependentvasoconstriction and increased corresponding shear stress (more than 25 dyn/cm2). Bosentan (a nonselectiveantagonist of ET-1 receptors) inhibits flow-dependent vasoconstriction induced by thecombination L-NAME/rHu-EPO without correcting vasodilation. Treatment of ET-1 transgenic micewith r-HuEPO increases systolic blood pressure, ET-1 plasma concentration, oxidative stress, infiltrationof aortic monocytes and macrophages (MOMA-2), pro-inflammatory cytokines levels INF-γ, TNF-α andIL-6 and exacerbate endothelial dysfunction. Exercise prevents all the deleterious effects of r-HuEPO. Itis also noteworthy that exercise increases the number of Foxp3-positive cells in the spleen and renalcortex. Then, the second aim of our study was to test the effect of adoptive transfer of Treg cells on HTNand vascular disorders induced in mice treated with angiotensin II (Ang II) or aldosterone (Aldo). Ourresults show that adoptive transfer of Treg prevents Ang II induced hypertension, endothelialdysfunction, prevents stiffness of mesenteric arteries, decreases oxidative stress and plasma levels ofpro-inflammatory cytokines (IFN-γ, TNF-α and IL-6) and the aortic and renal infiltration ofmacrophages. Adoptive transfer of Treg prevents the increase in systolic blood pressure induced byAldo, prevents impaired endothelium-dependent vasodilatation and hypertrophic remodelling ofmesenteric arteries, decreases oxidative stress and infiltration of inflammatory immune cells. Theadoptive transfer of T effectors cells (Teff) exacerbates the majority of the Aldo effects. Our resultsjustify, on one hand, the importance of exercise as a preventive tool for hypertension induced by r-HuEPO, and on the other hand, highlight the role of Treg as an essential component in the modulation ofhypertension and cardiovascular disorders.

No description available.

Erosion of a river bed has important implications with respect to scour around river structures such as bridges, transport of contaminants attached to the sediment, and disruption or destruction of aquatic habitats. Erosion occurs when the resistive strength of the sediment is overcome by the hydrodynamic forces produced by the flow of water. This resistance to erosion in a sediment originates from gravity or interparticle forces for coarse sediment (sand and gravel) and fine sediment (silt and clay), respectively. Since the erosion of fine sediment depends on the combination of many interparticle forces, and this combination fluctuates widely amongst different fine sediments, past studies have had difficulty finding a consistent method to estimate fine sediment erosion. This study analyzes sediments that fall in the transition size range between fine and coarse sediments and compares the findings with those from fine sediments (Wang 2013) and sandy coarse sediments (Navarro 2004, Hobson 2008), in order to correlate the erosion rates of both sediment types to their physical characteristics. In this study, kaolin-sand mixtures were prepared by mixing various percentages of Georgia kaolin by weight ranging from 30% to 100% with industrial fine sand and tap water. Geotechnical and other tests of sediment properties were performed to measure water content, bulk density, grain size distribution, temperature, pH, and conductivity of these mixtures. Hydraulic flume experiments measured the erosion rates of each sediment and these rates were used to estimate the critical shear stress correlating to that mixture. Relationships between the physical properties of the sediment and critical shear stress were developed by multiple regression analysis. An alternative option of estimating the critical shear stress by a weighted equation, which uses the combination of fine sediment erosion and coarse sediment erosion equations separately, was explored and found to be a viable and accurate option to estimating both coarse and fine sediment erosion from the same parameters and equation. The results from this study can be used to estimate sediment erodibility and thus river bed stability based on simple tests of physical properties of the river bed sediment and will help predict scour around bridges and other flow obstructions.

Master of Science / Department of Civil Engineering / Kyle Riding / The durability of partial depth concrete repair is directly related to the bond strength between the repair material and existing concrete. The wait time effects of cementitous grouts, epoxy, acrylic latex, and polyvinyl acetate bonding agents were observed on bond strength. Three rapid repair materials were as a comparison to bond strength, as well as concrete samples with no bonding agents having dry conditions and saturated surface dry moisture condition. The bonding agents and rapid repair materials were tested in a controlled laboratory environment. Bond strength loss with wait times of 0, 2, 5, 10, and 30 minutes were observed when bonding agents were applied. The laboratory samples were loaded using a direct shear test. Field tests were performed using the same repair materials and bonding agents. When the agents were applied in the field the wait times used were 0, 15, 30, and 45 minutes. 7 day and 5 month pull off tensile tests were performed during the field experiment. The data from both experiments show that when using cement grout bonding agents the high bond strength can be obtained when the repair material is applied within 15 minutes of application of the cement grout, and after 15 minutes bond loss can be expected. Wait time didn't have a significant effect on epoxy and acrylic latex bonding agents as long as they were placed before setting. The polyvinyl acetate agent and repair materials can develop high bond strength in laboratory settings, but when used in the field the bond strengths experience loss. When not using bonding agents in a repair, adequate bond strength can be obtained when using saturated surface dry condition.

In vitro mechanotransduction studies, uncovering the basic science of the response of cells to mechanical forces, are essential for progress in tissue engineering and its clinical application. Many varying investigations have described a multitude of cell responses, however as the precise nature and magnitude of the stresses applied are infrequently reported and rarely validated, the experiments are often not comparable, limiting research progress. This thesis provides physical and biological validation of a widely available fluid stimulation device, a see-saw rocker, as an In vitro model for cyclic fluid shear stress mechanotransduction. This allows linkage between precisely characterised stimuli and cell monolayer response in a convenient six-well plate format. Computational fluid dynamic models of one well were analysed extensively to generate convergent, stable and consistent predictions of the cyclic fluid velocity vectors at a rocking frequency of 0.5 Hz, accounting for the free surface. Validation was provided by comparison with flow velocities measured experimentally using particle image velocimetry. Qualitative flow behaviour was matched and quantitative analysis showed good agreement at representative locations and time points. A maximum shear stress of 0.22Pa was estimated near the well edge, and time-average shear stress ranged between 0.029 and 0.068Pa, within the envelope of previous musculoskeletal In vitro fluid flow investigations. The CFD model was extended to explore changes in culture medium viscosity, rocking frequency and the robustness to position on the rocking platform. Shear stress magnitude was shown to increase almost linearly with an increase in the viscosity of culture medium. Compared with 0.5 Hz, models at 0.083 and 1:167 Hz, the operational limits of the see-saw rocker, indicated a change in shear stress patterns at the cell layer, and a reduction and increase in mean shear stress respectively. At the platform edge at 0.5 Hz, a 1.67-fold increase in time-average shear stress was identified. Extensive biological validations using human tenocytes underlined the versatility of the simple In vitro device. The application of fluid-induced shear stress at 0.5 Hz under varying regimes up to 0.714Pa caused a significant increase in secreted collagen (p < 0.05) compared to static controls. Tenocytes stimulated at a shear stress magnitude of 1.023Pa secreted significantly less collagen compared to static controls. The potential for a local maximum in the relationship between collagen secretion rate and shear stress was identified, indicating a change from anabolic to catabolic behaviour. Collagen biochemical assay results were echoed with antibody stains for proteins, where a co-localisation of connexin-32 with collagen type-I was also identified. A custom algorithm showed that four hours of fluid-induced shear stress of 0:033Pa intermittently applied to tenocytes encouraged alignment and elongation over an eight day period in comparison to static controls. Primary cilia were identified in human tenocyte cultures and bovine flexor tendon tissue; however primary cilium abrogation In vitro using chloral hydrate proved detrimental to cell viability. Collaborative investigations identified that ERK signalling and c-Fos transcription factor expression peaked after the application of 0.012Pa at 0.083 Hz for 20 minutes and anabolic collagen gene expression relative quantities increased after 48 hours of rocking at 0.083 Hz. In conclusion, validated shear stresses within a six-well plate, induced by cyclic flow from a see-saw rocker, provides an exceptional model for the In vitro study of dynamic fluid shear stress mechanobiology. Biological investigations have been linked to precise applied shear stress, creating a foundation for understanding the complex relationship between tenocytes and fluid-induced shear stress In vitro. Using this model, research is repeatable, comparable and accurately attributed to shear stress, accelerating the scientific advancement of musculoskeletal mechanobiology.

High velocity channels are typically designed to discharge surplus water during severe flood events, and these types of flow are distinguished by high velocity, usually supercritical. A major challenge in high velocity channel design is to predict the free surface flow. Being able to predict the free surface flow profile beforehand can assist in selecting the best design for the channel as a whole. When the flow encounters a bridge pier, the streamline of the flow is separated and pressure may drop to a minimum; in contrast, velocity rises to its maximum value. As a result, cavitation damage may occur. The present study has used the computational fluid dynamics code ANSYS-CFX to investigate a full scale, three-dimensional engineering flow simulation of high velocity channels with different cross sections. The simulations were carried out on a high performance computing HPC cluster with 32 nodes. The code is based on the finite volume method and the Volume of Fluid (VOF) method was used to predict the position of the free surface profile. The impact of variation of the following parameters was investigated in terms of the free surface flow profile, both along the centreline and the wall of the channel: the minimum cavity index, and maximum shear stress on both bed and wall of the channel and on bridge pier; aspect ratio (channel bed width/flow depth), bed and side slopes of the channel, different discharges, which are represented by Froude numbers; the length and thickness of the bridge pier. First, the code sensitivity tools for convergence were examined. For this purpose, cases with different mesh sizes were examined and the best size chosen, depending on computation expense and convergence. Then, different turbulence models, such as the standard k-ε, RNG k-ε, and SST turbulence models were tested. The results show that the standard k-ε gives satisfactory results. Next, efforts were made to establish whether the flow achieved steady state conditions. This involved simulating two cases, one with steady state and the other with a transient state. Comparison of the two results shows that the flow properties do not change after three seconds and stay stable thereafter, so the flow can be considered as attaining a steady state. Finally, symmetry within the model geometry was tested, as this would allow a reduction in computation time, with only one side of the symmetrical model needing to be simulated. Two cases were investigated: firstly a simulation of only half of the channel geometry, and secondly a full geometry simulation. A comparison of the results of each case showed that the flow can be considered symmetrical along the centreline of the channel. Next, the code was validated against both numerical and experimental published results. For the free surface flow profile and velocity distribution the published experimental and numerical work of Stockstill (1996) was used; the ANSYS-CFX code results agree more closely with Stockstill’s experimental data than Stockstill’s numerical data. To test for shear stress distribution on the wall, uniform flow within a trapezoidal cross section channel was investigated and the results compared with those presented in the literature. The comparison shows good agreement between the ANSYS-CFX and published experimental works, for the predicted shear stress distributions on the walls and the bed of the channel. In total, sixty cases were simulated in order to investigate the impact of variations in the aforementioned parameters on maximum flow depth (both along the centreline and the wall of the channel) minimum cavity index, and maximum shear stress on both bed and wall of the channel and on bridge pier. Finally, non-dimensional curves are provided in addition to formulae derived from the data regression, which are intended to provide useful guidelines for designers.

Atherosclerosis and related cardiovascular diseases (CVDs) are amongst the largest causes of morbidity and mortality in the developed world, causing considerable monetary pressure on public health systems worldwide. Atherosclerosis is characterised by the build up of vascular plaque in medium and large arteries and is a direct precursor to acute vascular syndromes such a myocardial infarction, stroke or peripheral arterial diseases. The causative factors leading to CVD still remain relatively poorly understood, but are becoming increasingly identifiable as a dysfunction of the endothelial cells that line the arterial wall. It is well known that the endothelium responds to the prevailing fluid mechanic (i.e. haemodynamic) environment, which plays a crucial role in the localised occurrence of atherosclerosis near vessel bends and bifurcations. In these areas, disturbed haemodynamics lead to flow separation and very low wall shear stress (WSS), which directly affects the functionality of the endothelium and impedes the transport of important blood borne agonists and antagonists. Detailed full field measurements assessing complex haemodynamics are sparse and consequently this thesis aims to address some of the important questions related to arterial haemodynamics and CVD by performing in-vitro flow measurements in physiologically relevant conditions. In particular, this research develops and uses state-of-the-art Particle Image Velocimetry (PIV) techniques to measure three-dimensional velocity and WSS fields in scaled models of the human carotid artery. For this purpose, the necessary theoretical and experimental concepts are developed and in-depth analyses of the underlying factors affecting the local haemodynamics and their relation to CVD are carried out. In the first part, a methodology for the construct of transparent hydraulic flow phantoms from medical imaging data is developed. The arterial geometries are reproduced in optically clear silicone and the flowing blood is modelled with a refractive index matched blood analogue. Subsequently, planar and Stereo-PIV techniques are developed and verified. A novel interfacial PIV (iPIV) technique is introduced to directly measure WSS by inferring the velocity gradient from the recorded particle images. The new technique offers a maximal achievable resolution of 1 pixel and therefore removes the resolution limit near the wall usually associated with PIV. Furthermore, the iPIV performance is assessed on a number of numerical and experimental test cases and iPIV offers a significantly improved measurement accuracy compared to more traditional techniques. Subsequently, the developed methodologies are applied in three studies to characterise the velocity and WSS fields in the human carotid artery under a number of physiological and experimental conditions. The first study focuses on idealised vessel geometries with and without disease and establishes a general understanding of the haemodynamic environment. Secondly, a physiological accurate vessel geometry under pulsatile flow conditions is investigated to provide a more realistic representation of the true in-vivo flow conditions. The prevailing flow structure in both cases is characterised by flow separation, strong secondary flows and large spatial and temporal variations in WSS. Large spatial and temporal differences exist between the different geometries and flow conditions; spatial variations appear to be more significant than transient events. Thirdly, the three-dimensional flow structure in the physiological carotid artery model is investigated by means of stereoscopic and tomographic PIV, permitting for the first time the measurement of the full 3D-3C velocity field and shear stress tensor in such geometries. The flow field within the model is complex and three-dimensional and inherently determined by the vessel geometry and the build up of an adverse pressure gradient. The main features include strong heliocoidal flow motions and large spatial variations in WSS. Lastly, the physiological implications of the current results are discussed in detail and reference to previous work is given. In summary, the present research develops a novel and versatile PIV methodology for haemodynamic in vitro studies and the functionality and accuracy is demonstrated through a number of physiological relevant flow measurements.