This thesis focuses on the dynamic and static behaviour of reinforced concrete beams built using high-strength concrete and high-strength steel reinforcement. As part of this study, a total of 8 high-strength concrete beams, built with and without steel fibres, and reinforced with high strength ASTM A1035 bars are tested under simulated blast loading using the University of Ottawa shock-tube, with an additional 3 companion beams tested under quasi-static loading. The variables considered in this study include: concrete type, fibre content, steel reinforcement ratio and steel reinforcement type. The behaviour of the beams with high-strength steel bars is compared to a companion set of beams reinforced with conventional steel reinforcement. The criteria used to evaluate the blast performance of the beams includes: overall blast capacity, maximum and residual displacements, secondary fragmentation and crack control. The dynamic results show that high strength concrete beams reinforced with high-strength steel are able to resist higher blast loads and reduce displacements when compared to companion beams with conventional steel reinforcement. The results also demonstrate that the addition of steel fibres is effective in controlling crack formation, minimizing secondary blast fragments, reducing displacements and further increasing overall blast capacity. However, the use of high-strength steel and high-strength concrete also shows potential for brittle failures under extreme blast pressures. The static results show that specimens with high-strength steel bars do not increase beam stiffness, but significantly increase peak load carrying capacity when compared to beams with the same ratio of conventional steel reinforcement. The analytical research program aims at predicting the response of the test beams using dynamic inelastic single-degree-of-freedom (SDOF) analysis and includes a sensitivity analysis examining the effect of various modelling parameters on the response predictions. Overall the analytical results demonstrate that SDOF analysis can be used to predict the blast response of beams built with high-strength concrete and steel reinforcement with acceptable accuracy.

Selectins are key adhesion molecules responsible for initiating a multistep process that leads a cell out of the blood circulation and into a tissue or organ. The adhesion of cells (expressing ligands) to the endothelium (expressing the selectin i.e.,E-selectin) occurs through spatio-temporally regulated interactions that are mediated by multiple intra- and inter-cellular components. The mechanism of cell adhesion is investigated primarily using ensemble-based experiments, which provides indirect information about how individual molecules work in such a complex system. Recent developments in single-molecule (SM) fluorescence detection allow for the visualization of individual molecules with a good spatio-temporal resolution nanometer spatial resolution and millisecond time resolution). Furthermore, advanced SM fluorescence techniques such as Förster Resonance Energy Transfer (FRET) and super-resolution microscopy provide unique opportunities to obtain information about nanometer-scale conformational dynamics of proteins as well as nano-scale architectures of biological samples. Therefore, the state-of-the-art SM techniques are powerful tools for investigating complex biological system such as the mechanism of cell adhesion. In this project, several constructs of fluorescently labeled E-selectin will be used to study the conformational dynamics of E-selectin binding to its ligand(s) using SM-FRET and combination of SM-FRET and force microscopy. These studies will be beneficial to fully understand the mechanistic details of cell adhesion and migration of cells using the established model system of hematopoietic stem cells (HSCs) adhesion to the selectin expressing endothelial cells (such as the E-selectin expressing endothelial cells in the bone marrow).

Rheumatoid arthritis (RA) is a chronic, inflammatory autoimmune disease that targets joints, resulting in in permanent disability. Synovial macrophages have been implicated in the pathogenesis of RA; however, their exact origins and functions remains unclear. In this study, we show evidence that synovial macrophages are mostly derived from embryonic origin during normal development. Macrophages are derived from either hematopoietic stem cells (HSC) or erythro-myeloid progenitors (EMP), and it is postulated that different subpopulations of synovial macrophages may have distinct functions contributing to either homeostasis or inflammation. To investigate the phenotypes of synovial macrophage populations and characterize their lineage-specific functions in arthritic joints, we utilized both cell lineage-tracing and K/BxN serum-transfer arthritis mouse models. Utilizing Flt3Cre;Rosa26LSL-YFP mice to label HSC-derived cells, we demonstrated that there is minimal HSC contribution to synovial macrophage populations during homeostasis. Use of RankCre;Rosa26LSL-YFP and Cx3cr1CreERT2;Rosa26LSL-tdTomato mice to label EMP-derived cells corroborated the finding that the EMP compartment maintains the largest contribution to synovial macrophage populations during normal development. Analysis of macrophages in Csf1rMericreMer;Rosa26-LSLtdTomato mice provided definitive prove that synovial macrophages derived from yolk-sac EMP precursors in adult mice. Use of serum transfer arthritis (STA) mice demonstrated that while most macrophages in the inflamed synovium were EMP-derived, there was a marked increase in HSC-derived cells compared to those present in homeostasis. Although this study has contributed to eluding that the heterogeneity of synovial macrophages in both homeostasis and inflammatory arthritis (IA) is complex and lineage-specific, further studies are needed to clearly define lineage-specific functions of macrophages in synovial tissues and in IA.

Les dommages de l'ADN peuvent s’accumuler dans les cellules souches hématopoïétiques(CSH) suite aux stress externes ou métaboliques et perturber leur fonctionnement et/ou leur maintien.La réparation par excision de nucléotides (NER), initiée par l’arrêt de la transcription (TCR) ou par lareconnaissance de distorsions des régions non transcrites (GGR) de l’ADN, est nécessaire àl’hématopoïèse à long terme. XPC, un facteur clé du système GGR, participe à d’autres réponses austress oxydatif. Le laboratoire a montré que la perte de XPC provoque l’accumulation de mutations, unstress métabolique et la carcinogenèse. Notre objectif est d’évaluer son expression et son rôle dans lemaintien et la différenciation des CSH. Nos résultats montrent qu’il est plus exprimé dans les cellulesimmatures CD34+ que dans les CD34- matures. Aussi, XPC apparaît sous trois poids moléculairesdifférents certainement liés à des modifications post-traductionnelles. Son extinction par ARNinterférence n'affecte ni la prolifération ni la capacité progénitrice in vitro des cellules CD34+.Cependant, les cellules déficientes implantées chez des souris immunodéficientes disparaissentprogressivement suggérant une perte des CSH ou de leur capacité de différenciation. Postulant queles mutations s’accumulent avec le temps, nous avons étudié l’hématopoïèse chez des sourisdéficientes en XPC jeunes et âgées. Les différences décrites dans l’hématopoïèse chez les individusjeunes et âgés sont retrouvées mais, de manière surprenante, aucune différence entre les animauxsauvages et mutés quelque soit l’âge ou le stress génotoxique n’est observée. Les résultats obtenussur les cellules humaines démontrent un rôle potentiel de XPC dans l’hématopoïèse, mais denouvelles investigations sont nécessaires pour mieux comprendre les mécanismes impliqués, et lapossible participation de XPC dans la leucémogenèse. / DNA damage may accumulate in hematopoietic stem cells (HSC) due to external ormetabolic stresses, leading to perturbation in their function and/or maintenance. Nucleotide excisionrepair (NER), initiated in the DNA by the stop of transcription (TCR) or by the recognition of distortionsin transcribed regions (GGR), is necessary for long-term hematopoiesis. XPC, a key factor in GGR, isimplicated in oxidative stress. The laboratory has demonstrated that XPC loss leads to theaccumulation of mutations, metabolic stress and carcinogenesis. Our objective is to evaluate XPCexpression and its role in HSC maintenance and differentiation. Results showed that XPC is highlyexpressed in immature CD34+ cells compared to mature CD34- cells. In addition, XPC appeared withthree different molecular weights, certainly linked to post-translational modifications. XPC silencing byshRNA did not affect the proliferation or the progenitor ability of CD34+ cells in vitro. However, deficientcells transplanted in immunodeficient mice disappeared progressively, suggesting the loss of HSCs ortheir differentiation capacity. Postulating that mutations accumulate with time, we have studiedhematopoiesis in young and aged XPC deficient mice. Differences described in young and agedhematopoiesis systems were found but, surprisingly, no difference was observed between wild typeand mutant mice at any age or genotoxic stress. Data from human cells demonstrate a potential rolefor XPC in HSC but new investigations are necessary to better understand the mechanisms implicatedand if XPC may participate in leukemogenesis.

Since decades, hematopoietic stem cell transplantation (HSCT) has become a well established treatment modality for hematological malignancies and non-malignant disorders. Autologous and allogeneic hematopoietic stem cells (HSCs) mobilized into the peripheral blood (PB) have been used as a preferred source of transplantable stem cells1-3. And umbilical cord blood (UCB) has been introduced as a more attractive HSC source for HSCT, because fetal stem cells in UCB are speculated to be more primitive in comparison to adult stem cells. However the limited amount of HSCs is limiting their application for stem cell therapy in clinic. Therefore, people started to utilize extra-embryonic tissue to harvest more fetal stem cells, while people also tried to optimize the clinical protocol to mobilize more adult stem cells out of adult bone marrow. The innovative strategies and feasible procedures were discussed in this thesis. The axis of the chemokine receptor CXCR4 and its ligand SDF-1 is important for trafficking and homing of HSCs. It has already been demonstrated that the bicyclam AMD3100, a CXCR4 antagonist, in combination with G-CSF is able to induce a significant mobilization of CD34+ cells4. And human placenta is a potent hematopoietic niche containing hematopoietic stem and progenitor cells throughout development5. The homing of HSCs to the placenta is probably also mediated by the expression of SDF-1 as demonstrated for the bone marrow niche. In this study (part 1 of the chapter “Results and discussions”), we utilized AMD3100 to mobilize HSCs from placenta. And we can demonstrate that the CXCR4 antagonist AMD3100 mobilise placenta derived CD34+ cells ex utero already after 30 min of incubation and may further enhance the efficacy of harvesting placenta-derived HSC. The alpha4 integrin CD49d is involved in migration and homing of hematopoietic stem cells (HSC). Therapeutic application of natalizumab, an anti-CD49d antibody, in patients with multiple sclerosis (MS) has been associated with increased levels of circulating CD34+ progenitors. In our study (part 2 of the chapter “Results and discussions”), we compared circulating HSCs from MS patients after natalizumab treatment and HSCs mobilized by G-CSF in healthy volunteers, with regard to their migratory potential, clonogenicity and gene expression. CD34+ cells in the blood and marrow of natalizumab-treated patients expressed less of the stem cell marker CD133, were enriched for erythroid progenitors (CFU-E) and expressed lower levels of adhesion molecules. The level of surface CXCR-4 expression on CD34+ cells from patients treated with natalizumab was higher compared to that of CD34+ cells mobilized by granulocyte-colony stimulating factor (G-CSF) (median 43.9% vs. 15.1%). This was associated with a more than doubled migration capacity towards a chemokine stimulus. Furthermore, CD34+ cells mobilized by natalizumab contained more m-RNA for p21 and less MMP9 compared to G-CSF mobilised HSC. Our data indicate that G-CSF and CD49d blockade mobilize different HSC subsets and suggest that both strategies may be differentially applied in specific cell therapy approaches. In order to further improve the clinical outcome of HSC transplantation, many groups are focusing on ex vivo maintain or expand HSC. Unfortunately, the maintenance of HSC in vitro is difficult to achieve because of their differentiation. This is presumably caused by a lack of appropriate cues that are provided in vivo by the microenvironment. Indeed, HSCs located in the bone marrow are interacting with a specific microenvironment referred to as the stem cell niche, which regulates their fate in terms of quiescence, self-renewal and differentiation. An orchestra of signals mediated by soluble factors and/or cell-to-cell contact keeps the balance and homeostasis of self-renewal, proliferation and differentiation in vivo. To investigate the communication between HSCs and the niche, coculture assays with mesenchymal stromal cells (MSCs) were performed in vitro. Here, we can demonstrate that cell-to-cell contact has a significant impact on hematopoietic stem cells expansion, migratory potential and stemness. In this study (part 3 of the chapter “Results and discussions”), we investigated in more detail the spatial relationship between hematopoietic stem cells and mesenchymal stromal cells during ex-vivo expansion. And we defined three distinct localizations of HSCs relative to MSC layer: (i) those in supernatant (non-adherent cells); (ii) cells adhering on the surface of mesenchymal stromal cells (phase-bright cells) and (iii) cells beneath the mesenchymal stromal cells (phase-dim cells). Our data suggest that the mesenchymal stromal cell surface is the dominant location where hematopoietic stem cells proliferate, whereas the compartment beneath the mesenchymal stromal cell layer seems to be mimicking the stem cell niche for more immature cells. Our data provide novel insight into the construction and function of three-dimensional HSC–MSC microenvironments. In summary, we provided a new method to isolate fetal stem cells from extra-embryonic tissue (i.e. placenta) in the first part, then we discussed an innovative strategy with CD49d blockade to improve clinical modality for adult stem cell mobilization in the second part, and finally we investigated HSC maintenance and expansion in vitro and provided feasible way to mimic HSC niche in vitro in the last part. This thesis contributes to HSC-based stem cell therapy in two aspects, i.e. 1) fetal and adult stem cell isolation holding great therapeutic potential for blood diseases; 2) ex vivo stem cell manipulation providing a valuable platform to model HSC niche regulation.

No description available.

High-strength steel (HSS) conforming to ASTM A1035 is becoming increasingly used in various structural applications, including in high-rise buildings and bridges. Due to their chemistry and manufacturing process, ASTM A1035 steel bars result in a combination of high tensile strength to yield ratio and varying levels of corrosion resistance. One potential application of ASTM A1035 bars is in the blast-resistant design of concrete structures, where their use can allow for reduced steel congestion, and increased blast resistance. Despite their high initial cost, stainless steel (SS) reinforcing bars are also seeing increased use in concrete construction. Solid stainless steel bars are referenced in ASTM A955, which is applicable to various stainless steel alloys. In addition to their inherent corrosion resistance, most stainless steel bars possess greater tensile strength, and importantly, exceptional ductility, when compared to ordinary steel reinforcement. This unique combination of strength and ductility makes SS bars well-suited for blast design applications. The overarching aim of this thesis is to gain better understanding of the blast behavior of RC flexural members designed with high-strength (HSS) and stainless steel (SS) reinforcement. This objective is achieved through a combined experimental and numerical research program. As part of the experimental research, a large set of beams, subdivided into three series, are tested under either quasi-static bending or simulated blast loads using the University of Ottawa shock-tube. Series 1 (HSC-HSS) and Series 2 (HSC-SS) aim at examining the effects of blast detailing (as recommended in modern blast codes,) on the quasi-static, blast and post-blast behaviour of high-strength concrete (HSC) beams reinforced with either ASTM A1035 high-strength bars (8 beams) or ASTM A955 stainless steel bars (16 beams). In addition to the influence of detailing, the effects of steel grade/type, steel ratio and steel fibers are also studied. Series 3 further studies the benefits of combining higher grade or higher ductility reinforcement, with more advanced ultra-high performance concrete (UHPC). This series includes 20 UHPC beams built with either ordinary, HSS or SS reinforcing bars (UHPC-NSS, UHPC-HSS and UHPC-SS). In addition to the effect of steel grade/type, concrete type, steel ratio and steel detailing are also studied. The results from Series 1 and 2 demonstrate the benefits of implementing high-strength and stainless steel reinforcement in HSC beams subjected to blast loads, where their use leads to increased blast capacity, reduced support rotations, and higher damage tolerance. The results further demonstrate the benefits of “blast detailing” on the ductility and resilience of such beams, under both static and blast loads. The results also show that the use of steel fibers can be used to relax blast detailing in the beams with high-strength or stainless steel by increasing the required tie spacing from d/4 to d/2. The results from Series 3 confirm that the use of UHPC in beams enhances flexural response (in terms of strength and stiffness), which in turn results in superior blast resistance. Conversely, the high bond capacity of UHPC makes such beams more vulnerable to bar fracture. Increasing the steel ratio is found to effectively increase the failure displacement and ductility of the UHPC beams. The use of high-strength steel is found to increase load capacity and blast resistance, while the use of stainless steel results in remarkable ductility, which further enhances beam response under blast loading. As part of the numerical research program, the static and blast responses of the test beams are simulated using either 2D or 3D finite element (FE) modelling, using software VecTor2 and LS-DYNA. The numerical results show that the 2D FE modelling using software VecTor2 can provide reliable predictions of the static and blast responses of the HSS or SS reinforced HSC beams built with varying detailing, in terms of load-deflection response, cracking patterns, failure mode, displacement time histories and dynamic reactions. Likewise, the 3D FE modelling using software LS-DYNA with appropriate modelling of UHPC (using the Winfrith Concrete or CSCM models) can well predict the blast responses of UHPC beams with ordinary, high-strength and stainless steel, in terms of displacement/load-time histories, damage and failure modes.

No description available.

Atualmente, nota-se um crescimento na fabricação de produtos utilizando moldes e matrizes contendo formas geométricas complexas. No entanto, a fabricação destes ferramentais onde se emprega, principalmente, operações de fresamento, não tem acompanhado esta evolução com a mesma velocidade. O advento dos sistemas CAD/CAM, máquinas CNC e da tecnologia de usinagem em altas velocidades (HSC) influenciaram positivamente a fabricação de superfícies complexas. Contudo, nota-se ainda ineficiências neste processo produtivo. A qualidade superficial após as operações de usinagem ainda não é suficiente para que estes ferramentais entrem diretamente na linha de produção. Assim, operações manuais de acabamento são exigidas, elevando o tempo e custo de produção, comprometendo a qualidade dimensional. Com a finalidade de aprimorar a fabricação de moldes e matrizes, este trabalho apresenta uma revisão bibliográfica sobre a usinagem HSC; um estudo sobre a força de usinagem para o fresamento de formas complexas e uma análise sobre a metodologia utilizada por sistemas CAD/CAM e CNC para gerar e interpretar programas NC que contenham movimentações complexas de ferramenta. Análises práticas foram realizadas em um centro de usinagem HSC, e os resultados indicam que as limitações tecnológicas atuais na cadeia CAD/CAM/CNC limitam a usinagem de formas complexas com altas velocidades, reduzindo a velocidade de avanço programada e que as características intrínsecas deste processo de remoção de material demonstrou ser bastante complexo, acarretando em constantes alterações na força de usinagem / In recent years the number of products containing free-form shapes produced by dies and molds have been growing noticeably. However, the milling process used to manufacture those dies and molds does not meet their requirements. The arrival of the CAD/CAM systems and the High Speed Cutting Technology (HSC) helps to improve this manufacturing process. Although to obtain the surface quality needed to meet the dies and molds requirements, a hand finishing still requires. It involves time, money and decrease the product quality. Technological limitations in the CAD/CAM/CNC chain limit the feed rate when milling free-form shape. It also has a negative effect on the surface roughness. Besides, this kind of milling still lacks scientific knowledge of the cutting process. With the aim to support the dies and moulds fabrication, this work presents an overview about HSC Technology; the cutting forces in a non-planar milling; the cutting tool deflection; a detailed description of the process chain involving the CAD/CAM/CNC systems and the methodologies used by those systems to generate and accomplish free-free tool-paths. Free-form milling experiments applying the HSC Technology were made to study the behavior of this process, and the outcomes are presented

The objectives of this project were to survey the flotation circuit of the Boliden concentrator, mass balance collected data and evaluate a scale-up model for laboratory flotation results. The model assumes that half of the recovery to cleaner middlings in a standard laboratory test would report to the final concentrate if it were done in closed circuit, as is the case in a full-scale plant. It has been used by Boliden Mineral AB since 1982 but its accuracy had not been studied since 1986. The model can be categorised as of open circuit type with scale-up factors. The project was based on a complex Ag-Au-Cu-Pb-Zn sulphide ore from the Kristineberg mine. Laboratory tests were done to produce concentrates of CuPb, Cu, Pb and Zn with pulp samples from the concentrator as feed material. The software HSC 9.3 was used to mass balance data from the plant survey. It was decided that the model would be deemed usable if it was able to predict the plant results with the same accuracy as in the survey of 1986. A simulated locked cycle test with split factors (Agar & Kipkie, 1978) was identified as an alternative scale-up model. The results showed that the model was able to predict the plant results with the same accuracy as in 1986. It was especially good at predicting grade and recovery of the main element in a concentrate. For example, it predicted an 18 % higher grade and 11 % lower recovery of Cu to the CuPb concentrate, while a 3 % lower grade and 11 % lower recovery of Zn was predicted to the Zn concentrate. The locked cycle model gave much worse predictions on grades, but more accurate recoveries. It was also better at predicting the behaviour of minor impurity elements such as As and Bi. A recommendation is to combine the two alternatives in a type of "mixed cycle" model. In this study, it would have predicted an 18 % higher grade and 7 % lower recovery of Cu to the CuPb concentrate, as well as a 3 % lower grade and 1 % higher recovery of Zn to the Zn concentrate compared with plant results. Such a model seems to give better figures, but should be put to the test on more samples and ores to confirm this belief. It could at the very least be used to check the reliability of results predicted by the current scale-up model. / Syftet med det här examensarbetet var att utföra en detaljprovtagning av flotationskretsen i Bolidens anrikningsverk, massbalansera data och sedan utvärdera en modell för uppskalning av resultat från laboratorieflotationer. Modellen antar att hälften av utbytet till returgodset i ett satsvis laboratorieförsök skulle rapportera till det slutliga koncentratet om det återcirkulerades, såsom i ett anrikningsverk. Den har använts av Boliden Mineral AB sedan 1982 men utvärderades senast 1986. Kategoriskt kan den ses som en uppskalningsmodell av typen öppen krets med skalfaktorer. Projektet baserades på en komplex Ag-Au-Cu-Pb-Zn sulfidmalm från gruvan i Kristineberg. Laboratorieförsök utfördes för att ta fram koncentrat av CuPb, Cu, Pb och Zn, med pulpprover från driften som utgångsmaterial. Programmet HSC 9.3 användes för att massbalansera datan från provtagningen. Det bestämdes att modellen skulle anses som godtagbar ifall den kunde förutspå driftresultatet med samma noggrannhet som 1986. Ett simulerat försök av typen sluten krets (Agar & Kipkie, 1978) identifierades som den mest intressanta alternativmodellen och även den utvärderades. Resultaten visade att modellen än idag ger godtagbara förutsägelser med samma noggrannhet som 1986. Modellen var särskilt bra på att förutspå halt och utbyte av den huvudsakliga metallen till dess eget koncentrat. Den förutspådde exempelvis en 18 % högre halt och 11 % lägre utbyte av Cu till CuPb-koncentratet, samt 3 % lägre halt och 11 % lägre utbyte av Zn till Zn-koncentratet. Den alternativa modellen gav sämre förutsägelser med avseende på halter, men bättre med avseende på utbyten. Den var bättre på att förutspå beteendet hos låghaltiga föroreningar såsom As och Bi. Rekommendationen är att kombinera de två modellerna till en "blandkretsmodell". I den här undersökningen hade ett sådant alternativ förutspått en 18 % högre halt och 7 % lägre utbyte av Cu till CuPb-koncentratet, samt 3 % lägre halt och 1 % högre utbyte av Zn till Zn-koncentratet jämfört med driftresultatet. En sådan modell tycks ge bättre förutsägelser, men bör testas på fler prover och malmtyper. Den borde åtminstone kunna användas för att kontrollera trovärdigheten hos resultaten förutspådda av den nuvarande modellen.