Background: All treatments, especially funded by social means, should be subject to cost-effectiveness studies to ensure that the best possible optimization decision it taken between different treatment options. Within the health care area this is a well-developed area. For odontology in general, and orthodontic treatment in specific, this is an unknown territory and cost-effectiveness studies are rare. Malocclusion can be treated with several different systems. The sales pitch from system providers for braces - such as self-ligated brackets (SLB) promises e.g. shorten treatment duration, shorter chairtime, fewer visits, but the purchase cost of SLB systems is higher. Due to lack of results from unbiased RCT there is no guidelines. It is up to the dentist to use the treatment method/system of their own choice. Purpose: The purpose of this thesis is to estimate the cost-effectiveness of SLB compared to conventional brackets (CB) to determine which of the two systems that should be considered as the preferred choice of treatment in a publicly funded system. Method: Through an incremental cost-effectiveness ratio (ICER) calculate the most costeffective system from a societal perspective which includes all costs. To estimate the health care cost, the Reference price list will be used as baseline for production cost for CB. With the use of secondary resources examine the cost driving components to estimate the production cost of SLB. To estimate the non-health care cost a matrix with downtime from work and travel costs was estimated accounting for travel to disclose the magnitude of these cost. The Outcome of the different treatment alternatives will be measured from different perspectives, such as state specific and general profiles. Treatment duration and foregone education will also be investigated. Conclusions: When all production costs for SLB has been adjusted it shows that the total cost of SLB is slightly cheaper. The reduced number of visits required, less chairtime and shorter treatment period are the positive sides of the higher purchase cost of the SLB. Most of the outcome results are not statistically significant, but there is a tendency, with better average values for SLB. SLB seems to be the cheaper and with more effective outcome though not to the extent the provider promises. The difference of treatment duration is minimal and has no significant advantage. The foregone education for the patient is not possible to quantify in monetary terms but could have significant impact depending on different geographical areas but will small differences between the systems. Hence, in lack of unbiased studies and just focusing on average values, SLB dominates CB and should be the preferred choice of treatment until new long-term studies has been published. / Bakgrund: Alla behandlingar, särskilt de som är offentligt finansierade, borde ha genomgått kostnadseffektivitetsbedömningar för att kunna fatta bästa möjliga beslut. Inom sjukhälsovård är detta ett välutvecklat område. Inom tandvård generellt, men kanske ortodonti i synnerhet, är detta ett outforskat område och kostnadseffektivitetsstudier är sällsynta. Malocklusion kan behandlas med flera olika system. Säljargument från olika tandställningsleverantörer, som t.ex. självligerande brackets (SLB) ger löften om t.ex. kortare behandlingsperiod, kortare tid i tandläkarstolen, färre besök men till en högre inköpskostnad. I brist på resultat från oberoende RCT saknas riktlinjer. Det är upp till den enskilda tandläkaren att välja den behandlingsmetod/system som hen finner bäst. Syfte: Syftet med uppsatsen är att undersöka kostnadseffektiviteten mellan SLB jämfört med konventionella brackets (CB) för att avgöra vilket av dessa två system borde anses vara föredraget behandlingsalternativ i ett offentligt finansierat system Metod: Med kostnadseffektivitetskvoten (incremental cost-effectiveness ratio, ICER) beräkna det mest kostnadseffektiva systemet från ett samhällsperspektiv som inkluderar alla kostnader. För att uppskatta direkta behandlingskostnaden kommer Referensprislistan att användas som produktionskostnadsbas. Med hjälp av sekundära källor som undersöker de kostnadsdrivande komponenterna, uppskatta produktionskostnaden för SLB. För att mäta de indirekta behandlingskostnaderna skapas en matris för att påvisa kostnaden med ställtid från arbete och resekostnader. Resultatet av de olika behandlingsalternativen kommer att mätas från olika perspektiv, såsom generella hälsoprofiler och tillståndsspecifika. Behandlingsperiod och missad utbildnings kommer också att undersökas. Slutsats: Efter det att produktionskostnaderna har blivit justerade visas att totalkostnaden för SLB är något billigare. Färre antal besök, kortare tid i behandlingsrummet är det positiva sidorna av det högre inköpspriset för SLB. De flesta av resultaten är inte statistiskt signifikanta men det finns en tendens mot bättre effekt med SLB. SLB tycks vara det billigare och mest effektiva systemet om än inte i den utsträckning som säljargumenten hävdar. Skillnaden i behandlingsperiod är minimal och har igen avgörande betydelse. Den missade utbildningen är svårligen kvantifierbar i monetära termer men kan ha en avgörande betydelse beroende på olika geografiska platser men med små skillnader mellan systemen. Följaktligen, i brist på oberoende studier och med ett fokus på medelvärde är SLB det dominerande valet över CB och borde vara det föredragna behandlingsalternativet fram till dess att nya långsiktiga forskningsrön har blivit publicerade.

This thesis investigates the opportunities and challenges of using repurposed electric car batteries, so called Second-life Battery (SlB), in a residential building as an energy storage. The performance of SlBs is compared to a First-life Battery (FlB) by identifying two potential scenarios and using a battery degradation model. The first scenario involved the batteries providing ancillary services to the grid, while in the second scenario the batteries were used for peak shaving. The battery degradation model is based on typical usage from the scenarios. The thesis findings indicate that the SlB degrades at a slower rate than the FlB, but can perform fewer cycles. Economic performance varies based on the application and initial assumptions. Both batteries proved to be profitable in the two scenarios studied, with some of the SlB configurations outperforming FlB configurations and vice versa. The yearly compensation received, when providing ancillary services with a 1000 kWh battery, ranged between 3-8 MSEK, while the yearly compensation received, when peak shaving with a 200-300 kWh battery, ranged between 20-35 thousand SEK. The main challenges identified included reduced lifespan, security risks, potential price increases, and space constraints. On the other hand, the main opportunities identified included potential price decreases and sustainability benefits such as carbon footprint reduction and grid stability. These results may provide valuable insights for informed decision-making regarding investments in FlBs and SlBs on the Swedish market.

O presente trabalho trata da manufatura direta de implantes de hidroxiapatita mediante a técnica de prototipagem rápida. A hidroxiapatita proveniente de estrutura óssea mineral bovina possui grande importância dentre os biomateriais devido a sua biocompatibilidade e grande similaridade com a estrutura óssea humana. A prototipagem rápida permite a obtenção de modelos com dimensões e formato miméticos ao do defeito ósseo do paciente e possibilita a obtenção de implantes com porosidade desejada e geometria complexas. Este trabalho tem como objetivo o estudo da obtenção de implantes por três diferentes técnicas de prototipagem rápida: usinagem a verde, 3DPrint e uma inovadora técnica de prototipagem rápida obtida pela polimerização da resina acrílica (fase líquida da barbotina) por luz ultravioleta presente em uma faixa da banda do espectro de luz emitido pelo laser azul (SLB). A diminuta pressão de conformação, típicas das técnicas de prototipagem rápida aditiva, exige pós altamente reativos para efetiva sinterização, dessa forma a hidroxiapatita obtida a partir do fêmur bovino foi calcinada e moída em moinhos de bolas e vibratório na busca de pós submicrométricos. A usinagem a verde de compactados foi considerada como padrão ouro da manufatura e foi aplicada para obtenção de corpos de prova cilíndricos, previamente conformados por prensagem isostática a 100 e 200 MPa e ensaiados por densificação, flexão, dureza e tenacidade à fratura. Introdução de partículas de alumina e whiskers de alumina foram ensaiados na busca de otimização das propriedades mecânicas. Para prototipagem rápida pelo método de SLB, misturas de hidroxiapatita e resina foram submetidas à emissão do laser azul com percurso direcionado em equipamento CNC, onde peças tridimensionais foram prototipadas e sinterizadas. A hidroxiapatita proveniente do osso bovino apresentou características de íons substitucionais de carbonato e após moagens em moinho de bolas seguido de vibratório foi obtida partículas de diâmetro médio equivalente de 0,35 µm que se mostraram reativas para promoção da sinterização. A sinterização a 1300 ºC-2h forneceu as melhores características cristalográfica e de propriedades mecânicas, resultando em corpos referências com hidroxiapatita sem decomposição em outros fosfatos de cálcio com 99,8 % da densidade, resistência mecânica à flexão de 100 MPa, dureza de 483 HV e tenacidade à fratura de 0,6 MPa.m1/2. A introdução de 1 vol% de alumina e whiskers de alumina elevaram a resistência mecânica do corpo para aproximadamente 140 MPa com tenacidade de 0,61 MPa.m1/2. Para a técnica 3DPrint aglomerados esféricos da composição cerâmica deve ter dimensões entre as malhas de 200 e 400 mesh (75 a 38 µm) e o binder deve ter resistividade superior à 290 kΩ, componentes reativos, emolientes, lubrificantes e estabilizantes. Na manufatura pela técnica SLB a incidência do laser com fluência de 170 mW.s/mm2 promoveu a cura da resina no diâmetro de 0,5 mm e profundidade próxima a 0,5 mm o que, permitiu a prototipagem de scaffolds com camadas de espessura de 0,5 mm resistente ao manuseio para remoção da barbotina não curada. / The study deals with the direct manufacturing of hydroxyapatite implants using rapid prototyping. Hydroxyapatite structure originated from bovine mineral bone has great importance among the biomaterials and biocompatibility due to its great similarity with the human bone structure. Rapid prototyping allows obtaining models with dimensions and shaping mimetics to the patient\'s bone defect and enables the achievement of implants with complex geometry and porosity desired. This study aims to obtain scaffolds for three rapid prototyping techniques: the green machining, 3DPrint and an innovative rapid prototyping technique, obtained by polymerization of acrylic resin (liquid phase of slurry) by ultraviolet light present in a range of the spectrum band a blue light emitted by the laser (SLB). The small conformation pressure, typical of additive rapid prototyping techniques requires highly reactive powders for effective sintering, and then hydroxyapatite obtained from bovine femur was calcined and milled in ball mill and vibratory mill for obtention of sub micrometrics powders. The green machining was considered as the standard of mechanical properties and was used to obtain cylindrical specimens, previously shaped by isostatic pressing at 100 and 200 MPa, and tested by densification, bending, hardness and toughness fracture. Introduction of particles of alumina and alumina whiskers were tested in search engine optimization of the mechanical properties. For rapid prototyping by the method of SLB, mixtures of hydroxyapatite and resin were subjected to blue laser emission directed in CNC equipment where three-dimensional pieces were prototyped and sintered. Hydroxyapatite derived from bovine bone showed characteristics of substitutional carbonate ions and non-stoichiometry, after grinding in a ball mill followed by vibratory mill, particles obtained was equivalent average diameter of 0.35 micrometers that proved reactive to promote sintering. Sintering at 1300 °C-2h gave the best crystallographic characteristics and mechanical properties, resulting in references bodies with hydroxyapatite without decomposition of other calcium phosphates with 99.8 % of density, flexural strength of 100 MPa, hardness of 483 HV and fracture toughness of 0.6 MPa.m1/2. The introduction of 1 vol% alumina and alumina whiskers increased the mechanical strength of the body with approximately 140 MPa and toughness fracture of 0.61 MPa.m1/2. For technical 3DPrint spherical agglomerates of ceramic composition should have dimensions between 200 and 400 mesh (75-38 µm) and the binder should have resistivity higher than 290 kΩ and components reactive, emollients, lubricants and stabilizers. In the manufacturing technique SLB incidence of the laser with a fluency of 170 mW.s/mm2 promoted curing of the resin of 0.5 mm in diameter and depth about 0.5 mm, which enabled the prototyping of scaffolds with thick layers 0.5 mm resistant to handle for removal of slurry uncured.

O presente trabalho trata da manufatura direta de implantes de hidroxiapatita mediante a técnica de prototipagem rápida. A hidroxiapatita proveniente de estrutura óssea mineral bovina possui grande importância dentre os biomateriais devido a sua biocompatibilidade e grande similaridade com a estrutura óssea humana. A prototipagem rápida permite a obtenção de modelos com dimensões e formato miméticos ao do defeito ósseo do paciente e possibilita a obtenção de implantes com porosidade desejada e geometria complexas. Este trabalho tem como objetivo o estudo da obtenção de implantes por três diferentes técnicas de prototipagem rápida: usinagem a verde, 3DPrint e uma inovadora técnica de prototipagem rápida obtida pela polimerização da resina acrílica (fase líquida da barbotina) por luz ultravioleta presente em uma faixa da banda do espectro de luz emitido pelo laser azul (SLB). A diminuta pressão de conformação, típicas das técnicas de prototipagem rápida aditiva, exige pós altamente reativos para efetiva sinterização, dessa forma a hidroxiapatita obtida a partir do fêmur bovino foi calcinada e moída em moinhos de bolas e vibratório na busca de pós submicrométricos. A usinagem a verde de compactados foi considerada como padrão ouro da manufatura e foi aplicada para obtenção de corpos de prova cilíndricos, previamente conformados por prensagem isostática a 100 e 200 MPa e ensaiados por densificação, flexão, dureza e tenacidade à fratura. Introdução de partículas de alumina e whiskers de alumina foram ensaiados na busca de otimização das propriedades mecânicas. Para prototipagem rápida pelo método de SLB, misturas de hidroxiapatita e resina foram submetidas à emissão do laser azul com percurso direcionado em equipamento CNC, onde peças tridimensionais foram prototipadas e sinterizadas. A hidroxiapatita proveniente do osso bovino apresentou características de íons substitucionais de carbonato e após moagens em moinho de bolas seguido de vibratório foi obtida partículas de diâmetro médio equivalente de 0,35 µm que se mostraram reativas para promoção da sinterização. A sinterização a 1300 ºC-2h forneceu as melhores características cristalográfica e de propriedades mecânicas, resultando em corpos referências com hidroxiapatita sem decomposição em outros fosfatos de cálcio com 99,8 % da densidade, resistência mecânica à flexão de 100 MPa, dureza de 483 HV e tenacidade à fratura de 0,6 MPa.m1/2. A introdução de 1 vol% de alumina e whiskers de alumina elevaram a resistência mecânica do corpo para aproximadamente 140 MPa com tenacidade de 0,61 MPa.m1/2. Para a técnica 3DPrint aglomerados esféricos da composição cerâmica deve ter dimensões entre as malhas de 200 e 400 mesh (75 a 38 µm) e o binder deve ter resistividade superior à 290 kΩ, componentes reativos, emolientes, lubrificantes e estabilizantes. Na manufatura pela técnica SLB a incidência do laser com fluência de 170 mW.s/mm2 promoveu a cura da resina no diâmetro de 0,5 mm e profundidade próxima a 0,5 mm o que, permitiu a prototipagem de scaffolds com camadas de espessura de 0,5 mm resistente ao manuseio para remoção da barbotina não curada. / The study deals with the direct manufacturing of hydroxyapatite implants using rapid prototyping. Hydroxyapatite structure originated from bovine mineral bone has great importance among the biomaterials and biocompatibility due to its great similarity with the human bone structure. Rapid prototyping allows obtaining models with dimensions and shaping mimetics to the patient\'s bone defect and enables the achievement of implants with complex geometry and porosity desired. This study aims to obtain scaffolds for three rapid prototyping techniques: the green machining, 3DPrint and an innovative rapid prototyping technique, obtained by polymerization of acrylic resin (liquid phase of slurry) by ultraviolet light present in a range of the spectrum band a blue light emitted by the laser (SLB). The small conformation pressure, typical of additive rapid prototyping techniques requires highly reactive powders for effective sintering, and then hydroxyapatite obtained from bovine femur was calcined and milled in ball mill and vibratory mill for obtention of sub micrometrics powders. The green machining was considered as the standard of mechanical properties and was used to obtain cylindrical specimens, previously shaped by isostatic pressing at 100 and 200 MPa, and tested by densification, bending, hardness and toughness fracture. Introduction of particles of alumina and alumina whiskers were tested in search engine optimization of the mechanical properties. For rapid prototyping by the method of SLB, mixtures of hydroxyapatite and resin were subjected to blue laser emission directed in CNC equipment where three-dimensional pieces were prototyped and sintered. Hydroxyapatite derived from bovine bone showed characteristics of substitutional carbonate ions and non-stoichiometry, after grinding in a ball mill followed by vibratory mill, particles obtained was equivalent average diameter of 0.35 micrometers that proved reactive to promote sintering. Sintering at 1300 °C-2h gave the best crystallographic characteristics and mechanical properties, resulting in references bodies with hydroxyapatite without decomposition of other calcium phosphates with 99.8 % of density, flexural strength of 100 MPa, hardness of 483 HV and fracture toughness of 0.6 MPa.m1/2. The introduction of 1 vol% alumina and alumina whiskers increased the mechanical strength of the body with approximately 140 MPa and toughness fracture of 0.61 MPa.m1/2. For technical 3DPrint spherical agglomerates of ceramic composition should have dimensions between 200 and 400 mesh (75-38 µm) and the binder should have resistivity higher than 290 kΩ and components reactive, emollients, lubricants and stabilizers. In the manufacturing technique SLB incidence of the laser with a fluency of 170 mW.s/mm2 promoted curing of the resin of 0.5 mm in diameter and depth about 0.5 mm, which enabled the prototyping of scaffolds with thick layers 0.5 mm resistant to handle for removal of slurry uncured.

Masters in Public Administration - MPA / There are often noted gaps and tensions arise between official government policies and what is implemented on the ground. The two theories that consistently argue antagonistically about the policy processes are the rational bureaucratic model also called the "top-down approach" as opposed to the incremental or bottom-up approach. This research was inspired by a bottom-up perspective and involves a detailed investigation of Lipsky's street level bureaucracy (SLB) theory. Over the years since Lipsky's research much international work has be done on the subject of discretion among policy implementers but in the South the focus has been more on top-down ideas such as increasing state capacity and monitoring workers and not on the Lipsky problematic. Hence, this study specifically focuses on studying the extent, intentions and uses of discretion. Intentions are look e at in a threefold manner: good, bad and conflicting, among nurses as SLBs in a public hospital in Cape Town, in view of the t wo conflicting approaches to policy implementation. I found that the discretion practices among nurses do 'more good than harm' as opposed to the view held by the top down approach. The study further revealed that discretion is also often conflicted or ambivalent. Moreover, it is often based on tacit collective understandings and practices among groups of nurses. This is one element that needs to be explored further in future studies in order to contribute to the body of knowledge. Notably, there appears to be a gap in South African literature on this vital subject area. 111

Este artículo consiste en analizar una nueva alternativa de control antisísmico de alta tecnología para la construcción de hospitales en el Perú, que en su normativa exige el uso de aislamiento de base para edificaciones esenciales. Para ello, se opta por la evaluación estructural de una edificación construida con aisladores sísmicos y la misma con implementación del sistema de disipación de energía SLB (Shear Link Bozzo). Los resultados analizados, para cada técnica estudiada de control antisísmico, son: derivas de entrepiso, aceleraciones de piso, balance energético y análisis de rotulas plásticas, en base a la filosofía de funcionalidad continua. / This article consists of analyzing a new high-tech anti-seismic control alternative for the construction of hospitals in Peru, which in its regulations requires the use of base insulation for essential buildings. For this, the structural evaluation of a building constructed with seismic isolators is chosen and the same with the implementation of the SLB (Shear Link Bozzo) energy dissipation system. The results analyzed for each anti-seismic control technique studied are: mezzanine drifts, floor accelerations, energy balance and analysis of plastic hinges, based on the philosophy of continuous functionality. / Trabajo de investigación

Background Idiopathic pulmonary fibrosis (IPF) is a devastating form of chronic lung injury of unknown aetiology characterised by progressive lung scarring. A diagnosis of definite IPF requires High Resolution Computed Tomography (HRCT) appearances indicative of usual interstitial pneumonia (UIP), or in patients with 'possible UIP' CT appearances, histological confirmation of UIP. However the proportion of such patients that undergo SLB varies, perhaps due to a perception of risk of biopsy and additive diagnostic value of biopsy in individual patients. We hypothesised that an underlying UIP pathological pattern may result in increased risk of death and aimed to explore this by comparing the risk of SLB in suspected idiopathic interstitial pneumonia, stratified according to HRCT appearance. Additionally we sought to determine the positive-predictive value of biopsy to diagnose IPF in patients with 'possible UIP HRCT' in our population. In patients with possible UIP who are not biopsied, the clinical value of bronchoalveolar lavage (BAL) is uncertain. We aimed to prospectively study the diagnostic and prognostic value of BAL differential cell count (DCC) in suspected IPF and determine the feasibility of repeat BAL and the relationship between DCC and disease progression in two successive BALs. We hypothesised that BAL DCC between definite and possible IPF was different and that baseline DCC and change in BAL DCC predicted disease progression. Alveolar macrophages (AMs) are an integral part of the lung's reparative mechanism following injury, however in IPF they contribute to pathogenesis by releasing pro-fibrotic mediators promoting fibroblast proliferation and collagen deposition. Expansion of novel subpopulations of pulmonary monocyte-like cells (PMLCs) has been reported in inflammatory lung disease. We hypothesised that a distinct AM polarisation phenotype would be associated with disease progression. We aimed to perform detailed phenotyping of AM and PMLCs in BAL in IPF patients. Several prognostic scoring systems and biomarkers have been described to predict disease progression in IPF but most were derived from clinical trial patients or tertiary referral centres and none have been validated in separate cohorts. We aimed to identify a predictive tool for disease progression utilising physiological, HRCT and serum biomarkers in a unique population of incident treatment naïve IPF patients. Methods Between 01/01/07 and 31/12/13, 611 consecutive incident patients with suspected idiopathic interstitial pneumonia (IIP) presented to the Edinburgh lung fibrosis clinic. Of these patients 222 underwent video-assisted thoracoscopic lung biopsy and histological pattern was determined according to ATS/ERS criteria. Post-operative mortality and complication rates were examined. Fewer than 2% received IPF-directed therapy and less than 1% of the cohort were lost to follow-up. Disease progression was defined as death or ≥10% decline in VC within 12 months of BAL. Cells were obtained by BAL and a panel of monoclonal antibodies; CD14, CD16, CD206, CD71, CD163, CD3, CD4, CD8 and HLA-DR were used to quantify and selectively characterise AMs, resident PMLCs, inducible PMLCs, neutrophils and CD4+/CD8+ T-cells using flow cytometry. Classical, intermediate and non-classical monocyte subsets were also quantified in peripheral blood. Potential biomarkers (n=16) were pre-selected from either previously published studies of IPF biomarkers or our hypothesis-driven profiling. Linear logistic regression was used on each predictor separately to assess its importance in terms of p-value of the associated weight, and the top two variables were used to learn a decision tree. Results Based on the 2011 ATS/ERS criteria, 87 patients were categorised as 'definite UIP', of whom 3 underwent SLB for clinical indications. IPF was confirmed in all 3 patients based on 2013 ATS/ERS/JRS/ALAT diagnostic criteria. 222 patients were diagnosed with 'possible UIP'; 55 underwent SLB, IPF was subsequently diagnosed in 37 patients, 4 were diagnosed with 'probable IPF' and 14 were considered 'not IPF'. In this group, 30 patients were aged 65 years or over and 25/30 (83%) had UIP on biopsy. 306 patients had HRCTs deemed 'inconsistent with UIP', SLB was performed in 168 patients. Post6 operative 30-day mortality was 2.2% overall, and 7.3% in the 'possible UIP' HRCT group. Patients with 'definite IPF' based on HRCT and SLB appearances had significantly better outcomes than patients with 'definite UIP' on HRCT alone (P=0.008, HR 0.44 (95% CI 0.240 to 0.812)). BAL DCC was not different between definite and possible UIP groups, but there were significant differences with the inconsistent with UIP group. In the 12 months following BAL, 33.3% (n=7/21) of patients in the definite UIP group and 29.5% (n=18/61) in the possible UIP group had progressed. There were no significant differences in BAL DCC between progressor and non-progressor groups. Mortality in patients with suspected IPF and a BAL DCC consistent with IPF was no different to those with a DCC inconsistent with IPF (P=0.425, HR 1.590 (95% CI 0.502 to 4.967)). There was no difference in disease progression in either group (P=0.885, HR 1.081 (95% CI 0.376 to 3.106)). There was no statistically significant difference in BAL DCC at 0 and 12 months in either group. There was no significant change in DCC between 0 and 12 month BALs between progressors and non-progressors. Repeat BAL was well tolerated in almost all patients. There was 1 death within 1 month of a first BAL and 1 death within 1 month of a second BAL; both were considered 'probably procedure-related'. AM CD163 and CD71 (transferrin receptor) expression were significantly different between groups (P < 0.0001), with significant increases in the IPF group vs non fibrotic ILD (P < 0.0001) and controls (P < 0.0001 and P < 0.001 respectively). CD71 expression was also significantly increased in the IPF progressor vs non-progressor group (P < 0.0001) and patients with high CD71 expression had significantly poorer survival than the CD71low group (P=0.040, median survival 40.5 and 75.6 months respectively). CD206 (mannose receptor) expression was also significantly higher in the IPF progressor vs non-progressor group (P=0.034). There were no differences in baseline BAL neutrophil, eosinophil or lymphocyte percentages between IPF progressor or non-progressor groups. The percentage of rPMLCs was significantly increased in BAL fluid cells of IPF patients compared to those with non-fibrotic ILD (P < 0.0001) and healthy controls (P < 0.05). Baseline rPMLC percentage was significantly higher in IPF progressors vs IPF non-progressors (P=0.011). Baseline BAL iPMLC:rPMLC ratio was also significantly different between IPF progressor and non-progressor groups (P=0.011). Disease progression was confidently predicted by a combination of clinical and serological variables. In our cohort we identified a predictive tool based on two key parameters, one a measure of lung function and one a single serum biomarker. Both parameters were entered into a decision tree, and when applied to our cohort yielded a sensitivity of 86.4%, specificity of 92.3%, positive predictive value of 90.5% and negative predictive value of 88.9%. We also applied previously reported predictive tools such as the GAP Index, du Bois score and CPI Index to the Edinburgh IPF cohort. Conclusions SLB can be of value in the diagnosis of ILD, however perhaps due to the perceived risks associated with the procedure, only a small percentage of patients undergo SLB despite recommendations that patients have histological confirmation of the diagnosis. Advanced age is a strong predictor for IPF, and in our cohort 83% of patients aged over 65 years with 'possible UIP' HRCT appearances, had UIP on biopsy. BAL and repeat BAL in IPF is feasible and safe (< 1.5% mortality). Of those that underwent repeat BAL, disease progression was not associated with a change in DCC. However, 22% of lavaged patients died or were deemed too frail to undergo a second procedure at 12 months. These data emphasise the importance of BAL in identifying a novel human AM polarisation phenotype in IPF. Our data suggests there is a distinct relationship between AM subtypes, cell-surface expression markers, PMLC subpopulations and disease progression in IPF. This may be utilised to investigate new targets for future therapeutic strategies. / Disease progression in IPF can be predicted by a combination of clinical variables and serum biomarker profiling. We have identified a unique prediction model, when applied to our locally referred, incident, treatment naïve cohort can confidently predict disease progression in IPF. IPF is a heterogeneous disease and there is a definite clinical need to identify 'personalised' prognostic biomarkers which may in turn lead to novel targets and the advent of personalised medicines.

Much of our knowledge of cellular biology arises from direct observation of active cellular functions. Tools and techniques have steadily developed over the past several hundreds of years to aid in our understanding and control of the nanoworld and are referred to as nanotechnologies. In the context of nanotechnology, DNA is not used as a carrier for genetic information (as it is in cell), but as a construction material. DNA offers unprecedented control over the construction of simplified biomimetic models for the study of biological processes. This thesis first introduces and defines the field of DNA nanotechnology, with particular emphasis on the interaction of snthetic DNA nanostructures with biological membranes. Inspired by the protein clathrin, three-fold symmetric DNA tile made of eight, short DNA strands and capable of polymerising is described and studied, with the aim to interact with and controllably bend a membrane bilayer. This structure presented challenges during construction so an enhanced three-armed DNA structure built with DNA origami was designed. The succesful assembly of a rigid and functionalisable nanostructure is described. This origami structure was polymerised into large constructs in solution and on a supported lipid membrane. The shape of the structure was modulated to vary its curvature and apply a bending force to a lipid vesicle when anchored to it. Following the conclusion of this study, we present the construction of a small, unique DNA structure for enhanced electron microscopy imaging in cell lysate. This project is part of a developing technique to couple the interaction specificity of dyes in super-resolution microscopy and the high-resolution output of electron microscopy. Finally, the optimisation procedures and recommendations for TEM imaging of samples of DNA origami and lipid structures are discussed.

Les membranes cellulaires sont impliquées dans de nombreux processus cellulaires : la diffusion des médicaments et des ions, la transduction des signaux, la génération d'énergie, le développement cellulaire (fusion et fission). Les bicouches phospholipides sont les principaux composants des membranes cellulaires, elles constituent une barrière dynamique protégeant les réactions biochimiques cellulaires. La détermination des propriétés biochimiques et mécaniques des bicouches lipidiques et leur évolution avec les conditions environnementales est nécessaire pour étudier la nature des processus cellulaires et l'influence des agents externes (résistance mécanique, perméabilité et réponse biologique). Pour mener de telles caractérisations, des modèles simplifiés de membrane biomimétique, tels que des bicouches lipidiques supportées (SLB), ont été développés. Parmi les techniques de caractérisation disponibles, la microscopie à force atomique (AFM) a été largement utilisée pour étudier l'organisation nanométrique des SLB dans des conditions physiologiques. AFM peut produire des images à la haute résolution et peut également être utilisé pour quantifier la résistance mécanique des SLB au moyen d'expériences de perforation. Pendant 30 ans, AFM a traversé de nombreux développements. Très récemment, le Mode circulaire AFM (CM-AFM) a été développé à l'Université de Technologie de Compiègne. CM-AFM est capable de générer un mouvement de glissement de la pointe AFM sur l'échantillon à une vitesse élevée, constante et continue et de mesurer les forces de frottement latéral rapidement et exactement simultanément avec les forces verticales. Pour la première fois, le CM-AFM sert à caractériser les échantillons biologiques dans des conditions physiologiques, ce qui permet de mesurer simultanément les forces de poinçonnage et de frottement en fonction de la vitesse de glissement. Il offre pour la première fois la capacité de décrire le comportement de friction des SLB en complément de la force de perforation. En raison du besoin important de mesure quantitative, l'optimisation du protocole CM-AFM a été effectuée en premier. Le protocole d'étalonnage du scanner a été établi avec succès pour assurer la précision de la vitesse de glissement. En outre, le protocole d'étalonnage des pointes, basé sur la méthode de Wedge et un échantillon rayé, est également conçu pour déterminer la constante d'étalonnage de la force latérale. Nous avons utilisé CM-AFM pour mesurer les propriétés tribologiques des échantillons solides pour améliorer l'équipement sous milieu liquide. Ensuite, les propriétés mécaniques (forces de poinçonnage et de frottement) des SLB ont été mesurées en fonction de la vitesse de glissement. Les SLB purs et mixtes ont été préparés par la méthode de fusion des vésicules. Différents médias ont également été utilisés pour étudier l'effet des cations monovalents sur les propriétés mécaniques des SLB. Dans tous les cas, la force de frottement augmente linéairement avec la vitesse de glissement, ce qui nous permet de déduire le coefficient visqueux de frottement. Comme prévu, la force de poinçonnage et le coefficient visqueux de frottement sont influencés par la composition des mélanges de lipides, par la nature des cations en milieu liquide et par la longueur des chaînes hydrocarbonées mais pas de manière similaire. L'interprétation de l'évolution du coefficient de force de frottement visqueux avec le système étudié est particulièrement délicate car la force de frottement pourrait être influencée par les propriétés d'interface ou de volume. Cette problématique sera le défi pour les prochaines études. Néanmoins, nos résultats illustrent la puissance de la technique CM-AFM et ouvre de nombreuses possibilités pour caractériser d'autres échantillons biologiques (cellules et tissus) afin de mieux comprendre les mécanismes élémentaires de friction. / Cell membranes are involved in many cellular processes: drugs and ions diffusion, signal transduction, energy generation, cell development (fusion and fission). Phospholipid bilayers are the main components of cell membranes, they act as a dynamic barrier protecting cellular biochemical reactions. The determination of biochemical and mechanical properties of lipid bilayers and their evolution with environmental conditions is necessary to study the nature of cellular processes and the influence of external agents (mechanical resistance, permeability, and biological response). To conduct such characterizations, simplified biomimetic membrane models, such as supported lipid bilayers (SLBs), were developed. Among the available characterization techniques, atomic force microscopy (AFM) has been widely used to study the nanoscale organization of SLBs under physiological conditions. AFM can yield high resolution images and it can also be used to quantify the mechanical resistance of SLBs by means of punch through experiments. For 30 years, AFM has been through many developments. Very recently, the Circular Mode AFM (CM-AFM) has been developed at the Université de Technologie de Compiègne. CM-AFM is able to generate a sliding movement of the AFM tip on the sample at high, constant and continuous velocity and to measure the lateral friction forces fast and accurately simultaneously with the vertical forces. For the first time CM-AFM is used to characterize biological samples under physiological conditions, allowing the simultaneous measurement of both the punch-through and the friction forces as a function of the sliding velocity. It offers for the first time the ability to describe the friction behavior of SLBs in complement of the punch-through force. Due to the important need for quantitative measurement, optimization of the CM-AFM protocol has been done first. Protocol of scanner calibration has been successfully established to ensure the accuracy of sliding velocity. Besides, the protocol for tip calibration, based on wedge method and a scratched sample, is also made to determine the lateral force calibration constant. We have employed CM-AFM to measure the tribological properties of solid samples to improve the equipment under liquid medium. Then, the mechanical properties (punchthrough and friction forces) of SLBs were measured as function of the sliding velocity. Pure and mixed SLBs were prepared by the vesicle fusion method. Various media were also used to study the effect of monovalent cations to the mechanical properties of SLBs. In all cases, the friction force increases linearly with the sliding velocity allowing us to deduce the friction viscous coefficient. As expected both the punchthrough force and the friction viscous coefficient are influenced by the composition of lipid mixtures, by the nature of cations in liquid medium, and by the length of hydrocarbon chains but not in a similar fashion. The interpretation of the evolution of the viscous friction force coefficient with the studied system is particularly tricky as the friction force could be influenced by interface or volume properties. This problematic will be the challenge for the next studies. Nevertheless, our results illustrate how powerful the CM-AFM technique is and it opens wide opportunities to characterize other biological samples (cells and tissues) to gain a better understanding of the elementary mechanisms of friction.

L'Annexine A5 (Anx5) est une protéine de la famille des annexines qui présente la propriété de s'organiser à la surface des bicouches lipidiques en trimères et en assemblages bidimensionnels (2D) de trimères. Ce travail de thèse concerne le développement d'applications biotechnologiques des assemblages 2D d'Anx5 ainsi qu'une étude des propriétés fonctionnelles de ces assemblages. Ces travaux ont été réalisés principalement à l'aide des méthodes de la microbalance à cristal de quartz avec mesure de dissipation (QCM-D) et de la microscopie à force atomique (AFM). <br />La première partie de cette thèse concerne le développement de plates-formes d'ancrage basées sur des assemblages 2D de protéines dérivées de l'Anx5. Des complexes covalents entre l'Anx5 et des peptides d'adhésion cellulaire pour la réalisation de plates-formes d'ancrage de cellules. Des protéines de fusion entre l'Anx5 et un fragment de la protéine A ont d'autre part servi au développement de plates-formes 2D pour l'immobilisation contrôlée d'anticorps et de protéines. <br />La seconde partie de cette thèse concerne l'étude des propriétés fonctionnelles des assemblages 2D d'Anx5. L'étude de l'interaction d'un anticorps anti-phospholipide avec les assemblages 2D d'Anx5 montre que cet anticorps ne perturbe pas l'organisation 2D de l'Anx5. La seconde étude concernant l'influence des assemblages 2D de l'Anx5 sur la dynamique des bicouches lipidiques établie une corrélation entre la formation de trimères et le ralentissement de la dynamique des membranes.<br />Ces travaux contribuent à notre compréhension des propriétés des assemblages 2D d'Anx5 et permettent d'envisager le développement de biocapteurs pour cellules ou molécules.