GPR data interpretation is often difficult due to complex geological environments, reflections ambiguity and time-depth conversion uncertainty. Thanks to the geophysical fields interoperability it is possible to compare results of related survey methods. Radargrams from three model sites were processed and interpreted in an usual way to uniformly amplify all recorded reflections. Results were compared with ERT and EM (DEMP) data within integrated interpretation. GPR data interpretation was extended and new geological and geomorphological interpretations were uncovered. The GPR processing sequence was modified and simplified following outcomes from the integrated interpretation to ease different methods results comparison. Adapting a velocity model to precise time- depth conversion hasn't showed improvements. Key words: ground penetrating radar, electrical resistivity tomography, dipole electromagnetic profiling, integrated interpretation, integrated inversion, GPR processing optimization, sedimentary environment, velocity model

Mucopolysaccharidosis IIIB (MPS IIIB) is a rare, metabolic disorder that results from a deficiency in the lysosomal hydrolase, α-N-acetylglucosaminidase (Naglu). Naglu is a housekeeping enzyme involved in the degradation pathway of heparan sulfate. A deficiency in active Naglu leads to an accumulation of heparan sulfate within the lysosome, initiating a pathological cascade within the cell. Patients with MPS IIIB experience progressive central nervous system degeneration and die within the first few decades of life. Presently, enzyme replacement therapy, which is a standard of care for other lysosomal storage disorders, is an ineffective treatment for MPS IIIB. This is due to impermeability of the blood-brain barrier (BBB) to exogenous recombinant enzymes. A promising approach to this therapeutic obstacle is protein transduction domains. Protein transduction domains have been shown to facilitate the delivery of active enzyme across the BBB in mice. Previously, our laboratory used Spodoptera frugiperda (Sf9) insect cell system to express human recombinant Naglu fused to a synthetic protein transduction domain (PTD4). The purpose was to use PTD4 to the facilitate the delivery of Naglu across biological membranes, including the blood-brain barrier. However, a missing stop codon following PTD4 limited its transducibility. The stop codon was re-introduced and the improved fusion enzyme, Naglu-PTD4X, was stably expressed in Sf9 cells. The overarching goal of this project is to create a large-scale production of human recombinant Naglu that has the potential to be used to treat the neuropathology of patients with MPS IIIB. This project used a three-step purification system to purify Naglu-PTD4X. Uptake of Naglu-PTD4X was assessed in MPS IIIB fibroblasts using a fluorogenic activity assay, immunoblotting, and immunocytochemistry. Our purification system was successful at purifying Naglu-PTD4X to homogeneity with a 26% yield and specific activity of 84,000 units/mg. An increase in Naglu activity was detected in MPS IIIB fibroblasts following incubation with Naglu-PTD4X. Future directions will focus on optimizing immunodetection and conducting BBB penetration studies in murine models. / Graduate / 2020-06-21

Cerebrovascular diseases are by far the largest causes of death in the UK, as well as one of the leading causes of adult disability. The brain's healthy function depends on a steady supply of oxygen, delivered through the microvasculature. Cerebrovascular diseases, such as stroke and dementia, can interrupt the transport of blood (and hence oxygen) rapidly, or over a prolonged period of time. An interruption in flow can lead to ischaemia, with prolonged interruptions leading to tissue death and eventual brain damage. The microvasculature plays a key role in the transport of oxygen and nutrients to brain tissue; however, its role in diseases such as dementia is poorly understood, primarily due to the inability of current clinical imaging techniques to resolve microvessels, and due to the complexity of the underlying microvasculature. Therefore, in order to understand cerebrovascular diseases, it is necessary to be able to resolve and understand the microvasculature. In particular, generating large-scale models of the human microvasculature that can be linked back to contemporary clinical imaging is important in helping plug the current imaging gap that exists. A novel statistical model is proposed here that generates such large-scale models efficiently. Homogenization theory is used to generate a porous continuum capillary bed (characterised by its permeability) that allows for the efficient scaling up of the microvasculature. A novel order-based density-filling algorithm is then developed which generates morphologically accurate penetrating arterioles and venules, also demonstrating that the topology of the vessels only has a minor influence on CBF compared to diameter. Finally, the capillary bed and penetrating vessels are coupled into a large voxel-sized model of the microvasculature from which pressure and flux variations through the voxel can be analysed. A decoupling of the pressure and flux, as well as a layering of flow, was observed within the voxel, driven by the topology of the penetrating vessels. Micro-infarctions were also simulated, demonstrating the large local effects they have on the pressure and flux, whilst only causing a minor drop in CBF within the voxel.

Les peptides de la famille du LAH4 sont des peptides cationiques capables de se replier en hélice α amphiphile. Ces peptides sont riches en histidines ce qui permet de moduler les interactions de ces peptides de manière pH dépendante et dans une gamme physiologique. Leurs capacités à interagir et perturber les membranes ont été utilisées pour divers applications biologique, et notamment pour l'amélioration de systèmes de transport de gènes.Le travail de cette thèse a été divisé en trois parties dans le but de caractériser de manière biophysique les différentes interactions ayant lieu lors de la livraison du système de transport de gènes à l’intérieur d’une cellule. L’interaction peptide-peptide : avec l’étude de l’agrégation en fibrilles de la VF1 ; l’interaction peptide-membrane : avec l’effet du LAH4L1 en présence de membranes ; et l’interaction peptide-ADN : avec le suivit de l’interaction entre le LAH4L1 et de l'ADN. / The LAH4 family consists of cationic amphiphilic peptides with propensity to fold in α-helical secondary structures. They contain histidines allowing the modulation of their interactions in a pH dependent manner in the physiological range. In membranes, at neutral or acidic pH the peptide assumes a transmembrane or an in planar configuration, respectively.In the field of gene delivery systems, peptides like LAH4 are used. They are able to firstly interact with different cargoes in order to form stable complexes, then interact with the cell membrane, and finally, promote to escape from the endosome.This PhD has been divided into three parts in order to characterize, with biophysical methods, the interactions occurring during the delivery of these gene systems: peptide-peptide interactions with a focus on the study of VF1 fibre formation; peptide-membrane interactions: with the investigation of the effect of LAH4L1 in different membranes; and peptide-DNA interactions, where the interactions of LAH4L1 with a small DNA fragment were measured.

La thèse présentée décrit la préparation et l’étude d’auto-assemblages élaborés à partir de conjugués amphiphiles Tat-b-poly(triméthylène carbonate) (Tat-b-PTMC) doués de propriétés d’internalisation cellulaire conférées par le segment peptidique Tat. L’objectif principal de ces travaux était d’établir et de comprendre les liens entre la structure macromoléculaire, les caractéristiques colloïdales et l’activité biologique de ces systèmes. Les efforts de précision moléculaire et de caractérisation fournispour la synthèse des chimères Tat-b-PTMC a permis de corréler finement leurs structures chimiquesaux paramètres physico-chimiques des nanoparticules obtenues. Grâce à une approche expérimentale transverse combinant des études de biologie cellulaire et de biophysique, le mécanisme d’interaction in vitro de ces nanoparticules avec les cellules HeLa a pu être en partie élucidé. Enfin, un camouflage électrostatique pH-sensible a été mis au point pour tenter de moduler leur activité et d’augmenter leur sélectivité vis-à-vis de l’environnement tumoral. / This thesis work deals with preparation and study of cell-penetrating self-assemblies from amphiphilicpolymer-b-peptide Tat-b-poly(trimethylene carbonate) conjugates. Tat-b-PTMC chimeras withtunable hydrophilic fractions were synthesized, thoroughly characterized and self-assembled inaqueous buffer into size-tunable, highly monodisperse core-shell nanoparticles, presenting a full Tatcorona. Their physico-chemical profiles were assessed by complementary imaging (AFM, TEM) andscattering techniques (multiangle DLS, SANS) and correlated with their molecular architectures.Their transduction ability in vitro on HeLa cells and interaction mode with phospholipid membraneswere studied with a view to correlate their physico-chemical profiles with their biological properties.This interdisciplinary approach partially shed light on the interactions at play in the cellular uptakeprocess. With the ultimate goal of improving pharmacological characteristics, we finally endeavoredto develop an ON/OFF PEGylation strategy to harness the cell penetrating power of thosebiomacromolecular self-assembled systems.

Les travaux de cette thèse concernent l'étude d'une méthode d'évaluation des constantes diélectriques du béton d'ouvrage à risque par une approche problème inverse. Pour cela des mesures radar de type GPR et un modèle électromagnétique de ces mêmes mesures seront utilisés. Les deux premiers chapitres de la thèse présentent les concepts clés, le contexte et un historique des travaux déjà réalisés dans le domaine. Le chapitre 2 propose en outre une adaptation des travaux déjà réalisés à la recherche de gradients de propriétés diélectriques dans le béton en profondeur. Cette recherche préliminaire nous a permis de redéfinir le processus de mesures pour le rendre plus adapté à nos besoins. Ainsi le chapitre 3 présente une campagne de mesure réalisée à l'aide d'un nouveau dispositif radar sur des corps d'épreuve conditionnés de telle sorte que la répartition de la teneur en eau soit maîtrisée. Dans ce chapitre, nous détaillons également un nouveau modèle numérique 3D du dispositif qui nous permet de simuler nos mesures de façon réaliste Ces simulations sont nécessaires à l'expression du problème inverse. à partir de ces mesures, dans le quatrième chapitre, nous définissons tout d'abord le problème inverse, puis en utilisant des mesures sur les corps d'épreuves saturés, pouvant être considérés comme homogènes d'un point de vue diélectrique, nous résolvons ce problème inverse par un algorithme d'optimisation basé sur celui de Levenberg-Marquardt. Les résultats obtenus par notre processus d'inversion sont cohérents avec les valeurs physiques que l'on pouvait attendre et sont indépendants des valeurs données à nos inconnues lors de l'initialisation. Ces résultats correspondent effectivement à un minimum de la fonction coût qui mesure la distance entre mesures et simulations. Nous avons ensuite étudié des stratégies pour diminuer le temps de calcul de notre processus d'inversion. Une première approche consiste à utiliser un modèle simplifié en 2D du dispositif, qui permet d'initialiser l'inversion 3D avec un point plus proche de la solution. Une autre idée consiste à utiliser au mieux les performances d'une machine multiprocesseurs en parallélisant le code 3D. Dans le chapitre 5 nous nous intéressons à des mesures sur des corps d'épreuve en béton dont la teneur en eau est une fonction affine de la profondeur. Dans ces conditions nous avons fait l'hypothèse, en première approche, que les constantes diélectriques suivent aussi ce type de loi. Après avoir redéfini les inconnues de notre problème inverse et adapté l'algorithme, nous montrons que le processus d'inversion fourni des résultats cohérents en ce qui concerne la permittivité diélectrique, mais insatisfaisant pour la conductivité. Après analyse de ces résultats nous avons proposé diverses pistes pour améliorer et valider la méthode d'inversion. Finalement en comparant l'ensemble des résultats, il apparaît que notre méthode d'inversion permet de caractériser des gradients de permittivité, mais pas de conductivité. Cela représente néanmoins une avancée significative pour la mesure de la teneur en eau dans les bétons, facteur principal influençant la constante diélectrique de ce matériau, dans le contexte du contrôle non destructif des ouvrages du génie civil. / That thesis aims to propose an assessment method of the concrete electromagnetic properties by using radar measurements in a context of building sustainability, by inverse problem. For that purpose Ground Penetrating Radar (GPR) measurements are used as well as a numerical model of the radar device. The first two chapters of the thesis present the key concepts, the context of the study, as well as the history of the research on that topic in particular in our laboratory. Chapter 2 also expands those early works on the search of electromagnetic properties of concrete in depth. That preliminary work helped us to develop a measurement device that is more fitted to our needs. Chapter 3 presents a measurement campaign that was carried out by using a new measurement device on specially made reinforced concrete slabs whose water content is monitored. In that chapter a 3D model of the device is also described. That model allows us to simulate the measurements very precisely. Those simulations are a critical link on the inverse process. Chapter 4 details the formulation of the inverse problem for the case of concrete slabs saturated with water, which can be therefore considered to have homogeneous electromagnetic properties. The problem is solved by using an optimization algorithm based on Levenberg-Marquardt method. The values obtained by the inverse process are physically acceptable and independent of the values given to the unknowns when initializing the algorithm. Those values are the minimum of the cost function that computes the distance between measured and simulated electromagnetic fields. We then studied some strategies to reduce the computing time of the inverse process. A first approach consists in the use of a 2D simplified model of our device to initialize the 3D inversion with values closer to the searched solution. Another idea is to better take advantage of all the resources of a multiprocessor computer by implementing a parallel version of the 3D code. In chapter 5 we focus on the concrete slab with water content being an affine function of depth. We define the new parameters that represent the unknowns of our problem and we adapt the previous algorithm. Then we show that the inverse process gives satisfying results with regard to the dielectric permittivity only. We suggest some ways to improve the method. Finally we conclude that our method allows to retrieve the permittivity of the concrete, but not its conductivity. Nevertheless that result represents a significant step for the measuring of water content in reinforced concrete in particular when a gradient is present, by using radar measurements.

L’utilisation d’oligomères d’α-amino-γ-lactame (Agl-αAA) comme vecteurs de pénétration intracellulaire est décrite dans ce manuscrit. Nous avons montré que ces oligomères structurés en ruban sont capables de traverser la membrane plasmique pour atteindre le cytosol et y délivrer un cargo biologiquement actif. A la différence des séquences peptidiques, ces oligomères ont une très bonne résistance enzymatique. Une nouvelle famille de foldamères d’α-amino-γ-lactame (Agl-βAA) obtenus à partir de séquences /β peptidiques est également décrite. La structure secondaire de ces composés a été étudiée par RMN, IR-TF, CD et DRX. Nous avons montré que ces oligomères sont capables d’adopter une structure stable en hélice 12. De façon remarquable, ces oligomères sont solubles en milieux aqueux malgré une absence totale de chaînes latérales hydrophiles. / The use of α-amino-γ-lactam oligomers (Agl-αAA) as cell penetrating vectors are described in this work. These ribbon structured oligomers are able to cross the cell membrane to reach the cytosol and deliver a biologically active cargo. Unlike peptide sequences, these oligomers display a strong enzymatic resistance. A new family of α-amino-γ-lactam oligomers (Agl-βAA) obtained from conversion of /β peptide sequences are also described. Secondary structure of these molecules have been studied by NMR, FTIR, CD and XRD. These oligomers are able to adopt a stable 12-helix structure. Unexpectedly, these oligomers are soluble in aqueous mediums without any hydrophilic side chains

<p>This paper deals with archaeological prospection of an Iron Age farm site in Toland, Fresta parish, Uppland County in Sweden. The purpose of the paper has been to see whether the geophysical methods applied (GPR & EM-38) could produce useful results that could motivate its use in similar surveys in the future. Geochemistry has been used for the purpose of identifying possible activity areas on the site. The results have shown that it is possible with a GPR survey to identify postholes originating from the Migration Period longhouse at the site. The combining of several methods have been important for the identification and interpretation of several areas of interest.</p>

This paper deals with archaeological prospection of an Iron Age farm site in Toland, Fresta parish, Uppland County in Sweden. The purpose of the paper has been to see whether the geophysical methods applied (GPR &amp; EM-38) could produce useful results that could motivate its use in similar surveys in the future. Geochemistry has been used for the purpose of identifying possible activity areas on the site. The results have shown that it is possible with a GPR survey to identify postholes originating from the Migration Period longhouse at the site. The combining of several methods have been important for the identification and interpretation of several areas of interest.

This study presents a holistic view of which factors small and medium-sized enterprises (SMEs) have to consider, in moving their customer order decoupling point (CODP) further from the customer, thereby becoming more customized. The study also investigate whether SMEs and larger enterprises differ in the strategic questions relating to what influences an enterprise to become more customized, which prerequisites are required and which problems enterprises should anticipate and prepare for. These questions are important, given that strategic change is challenging, and can take considerable time to accomplish; it therefore needs to be based on the right decisions. This study is motivated by the fact that the markets SMEs are selling to tend to become more and more complex and harder to anticipate. Increased globalisation and use of IT and ICT have lead to greater uncertainty in customer demand. At the same time customers have ever more demanding needs and specifications; if these are not met, they will go elsewhere rather than reducing or adapting them. This complexity requires a more customized production and is a way to gain commercial advantages.       Customised production includes many benefits such as reduced uncertainty, by producing totally customized products less capital bound is tied up. But meanwhile many challenges such as reduced production efficiency, furthermore flexibility must be increased and higher demands on communications and IT systems are often necessary. The study starts with a wide and thorough theoretical overhaul around the movement of the CODP, but also addresses other parts of the organization to get the all-important big picture. This theoretical work is leading to a work-model which is used in six interviews within SMEs. In order to get a thorough picture, the enterprises that take part in the study are from different line of business and have from 18 to 235 employees. During these interviews, conducted on a single occasion per company, the model and related questions are discussed. Thereby an analysis of the interviews and the theory is done, with the model as the starting-point.   The result of this study is a basic model, which SMEs can use as an analytical tool, and a checklist, to facilitate a move towards a make-to-order manufacturing strategy, thereby gaining more flexibility. The study provides a basic generic framework which can be adapted and tailored by the company in question to see which parts are relevant to specific circumstances.