La gestion des maladies infectieuses dans la faune sauvage se heurte à de nombreuses limites, et le développement de stratégies efficaces représente un défi de taille. Pour atteindre cet objectif, une compréhension fine des facteurs influençant la transmission et la persistance de l’infection est nécessaire. Parmi ces facteurs, l’hétérogénéité de transmission est une caractéristique importante des populations sauvages. En effet, la diversité des comportements, des structures sociales et spatiales, ou encore des espèces peut conduire à des contributions très variables au nombre de nouvelles infections. Par conséquent, quantifier l’hétérogénéité de transmission pourrait permettre d’améliorer l’efficacité des mesures de gestion sanitaire dans la faune sauvage, en ciblant les individus ou les unités de population qui sont responsables de la majorité des évènements de transmission. L’objectif de cette thèse était d’améliorer les connaissances sur la gestion des maladies infectieuses dans des populations sauvages hétérogènes, en utilisant la brucellose à Brucella melitensis dans une population de bouquetin des Alpes (Capra ibex) comme modèle d’étude. En effet, la biologie de la brucellose et l’écologie de l’espèce hôte se prêtent bien à l’existence et donc à l’étude d’une hétérogénéité de transmission à différentes échelles. A l’aide de cultures bactériennes, nous avons tout d’abord montré que seulement 58 % des individus séropositifs sont à risque d’excréter la brucellose, et que ce risque diminue avec l’âge. Ensuite, mettant à profit l’existence d’informations détaillées sur la dynamique de population et le comportement du bouquetin, et de données épidémiologiques dans la population d’étude, nous avons développé un modèle individu-centré afin de quantifier l’hétérogénéité individuelle et spatiale de la transmission. Nous avons démontré que la transmission de la brucellose était hétérogène entre individus, les femelles provoquant environ 90% des nouvelles infections, et entre unités spatiales, plus de 80% des cas de transmission ayant lieu dans les trois sous-unités socio-spatiales qui forment la zone cœur du massif. Nous avons également estimé l’évolution temporelle de la séroprévalence et de la force d’infection, en utilisant différents modèles statistiques. Les résultats suggèrent que l’importante opération de capture menée en 2015, avec test systématique et élimination des individus séropositifs, a permis de diminuer la transmission de la brucellose dans la population. Sur la base de l’ensemble de ces résultats, nous avons évalués une série de stratégies de gestion sanitaire qui pourraient être utilisées à l’avenir dans la population. Les résultats, issus du modèle individu-centré, confirment que la stratégie prioritaire devrait être d’éliminer le plus d’individus séropositifs, et que cibler les femelles et/ou la zone cœur permet d’améliorer l’efficacité des mesures. Bien qu’il n’y ait pas de solution évidente pour la gestion de la brucellose dans notre cas d’étude, les stratégies de gestion ciblées sont très prometteuses et permettent de raffiner les mesures sanitaires classiquement utilisées. Il est donc primordial de bien comprendre l’hétérogénéité de transmission dans les populations sauvages infectées, et de rechercher des stratégies ciblées qui peuvent permettre d’améliorer la gestion en termes d’efficacité et d’acceptabilité / The management of infectious diseases in wildlife reservoirs is particularly challenging and faces several limitations. The development of appropriate management strategies requires a detailed understanding of the factors affecting the transmission and persistence of the infectious agent in the population. Among these factors, heterogeneity of transmission is a common characteristic in natural host-pathogen systems. Indeed, wild animals express a broad range of behaviours, are organised in a variety of social and spatial structures, occupy many areas with very different characteristics and belong to a large diversity of species. Such heterogeneities, from between-individuals to between-species, may result in different contributions to the overall number of new cases of infections. Thus, understanding transmission heterogeneity could provide valuable insights on how to effectively manage these systems, by targeting the individuals or areas that are responsible for most transmissions. The aim of this thesis was to provide insights on the monitoring and management of infectious diseases in heterogeneous wild populations, using Brucella melitensis infection in a French population of wild Alpine ibex (Capra ibex) as a case study. The biology of brucellosis and the ecology of Alpine ibex makes this case study a good candidate for transmission heterogeneity at several levels. Using bacterial examinations, we first established that only 58% of seropositive individuals were at risk to excrete Brucella, and that this risk decreased with increasing age. Then, we took advantage of detailed information available on ibex population dynamics, behaviour, and habitat use, and on epidemiological surveys, to build an individual-based model in order to quantify heterogeneity at the individual and spatial levels. The transmission is extremely heterogeneous between individuals, with females generating around 90% of the new cases of brucellosis infection, and between spatial units, three of the five socio-spatial units (the core area) accounting for more than 80% of brucellosis transmission. Using statistical models to estimate the temporal dynamics of the seroprevalence and of the force of infection in the population, we found evidence that the massive captures with test-and-remove operations that were conducted in 2015 managed to reduce brucellosis transmission in the population. Based on these results, we evaluated several predictive disease management strategies in the individual-based model. Our results confirmed that the primary strategy should be to remove as many seropositive individuals as possible, and that strategies targeting females and/or the core area are more effective than untargeted management. Although there is no silver bullet for the management of brucellosis in the population of study, targeted strategies offer a wide range of promising refinements to classical sanitary measures. We therefore encourage to look for heterogeneity in other infection-wildlife systems and to evaluate potential targeted strategies for improving management schemes in terms of efficiency and acceptability

L’interaction des particules chargées avec la matière conduit à un dépôt d’énergie très localisé dans des traces de dimensions sub-micrométriques. Cette propriété unique rend ce type de rayonnement ionisant particulièrement intéressant pour disséquer les mécanismes moléculaires radio-induits suite à l’échelle de la cellule. L’utilisation de microfaisceaux de particules chargées offre en outre la capacité d’irradier sélectivement à l’échelle du micromètre avec une dose contrôlée jusqu’à la particule unique. Mon travail a porté sur des irradiations réalisées avec le microfaisceau de particules chargées de la plateforme AIFIRA (Applications Interdisciplinaires des Faisceaux d’Ions en Région Aquitaine) du CENBG. Ce microfaisceau délivre des protons et particules alpha et est dédié aux irradiations ciblées in vitro (cellules humains) et in vivo (C. elegans).En complément de l’intérêt qu’elles présentent pour des études expérimentales, les dépôts d’énergie et les interactions des particules chargées avec la matière peuvent être modélisés précisément tout au long de leur trajectoire en utilisant des codes de structures de traces basés sur des méthodes Monte Carlo. Ces outils de simulation permettent une caractérisation précise de la micro-dosimétrie des irradiations allant de la description détaillée des interactions physiques à l’échelle nanométrique jusqu’à la prédiction du nombre de dommages à l’ADN et leurs distributions dans l’espace.Au cours de ma thèse, j’ai développée des modèles micro-dosimétriques basés sur l’outil de modélisation Geant4-DNA dans deux cas. Le premier concerne la simulation de la distribution d’énergie déposée dans un noyau cellulaire et le calcul du nombre des différents types de dommages ADN (simple et double brin) aux échelles nanométrique et micrométrique, pour différents types et nombres de particules délivrées. Ces résultats sont confrontés à la mesure expérimentale de la cinétique de protéines de réparation de l’ADN marquées par GFP (Green Fluorescent Protein) dans des cellules humaines. Le second concerne la dosimétrie de l’irradiation d’un organisme multicellulaire dans le cadre d’études de l’instabilité génétique dans un organisme vivant au cours du développement (C. elegans). J’ai simulé la distribution de l’énergie déposée dans différents compartiments d’un modèle réaliste en 3D d’un embryon de C. elegans suite à des irradiations par protons. Enfin, et en parallèle de ces deux études, j’ai développé un protocole pour caractériser le microfaisceau d'AIFIRA à l’aide de détecteurs de traces fluorescent (FNTD) pour des irradiations par protons et par particules alpha. Ce type de détecteur permet en effet de visualiser les trajectoires des particules incidentes avec une résolution de l’ordre de 200 nm et d’examiner la qualité des irradiations cellulaires réalisées par le microfaisceau. / The interaction of charged particles with matter leads to a very localized energy deposits in sub-micrometric tracks. This unique property makes this type of ionizing radiation particularly interesting for deciphering the radiation-induced molecular mechanisms at the cell scale. Charged particle microbeams (CPMs) provide the ability to target a given cell compartment at the micrometer scale with a controlled dose down to single particle. My work focused on irradiations carried out with the CPM at the AIFIRA facility in the CENBG (Applications Interdisciplinaires des Faisceaux d’Ions en Région Aquitaine). This microbeam delivers protons and alpha particles and is dedicated to targeted irradiation in vitro (human cells) and in vivo (C. elegans).In addition to their interest for experimental studies, the energy deposits and the interactions of charged particles with matter can be modeled precisely along their trajectory using track structure codes based on Monte Carlo methods. These simulation tools allow a precise characterization of the micro-dosimetry of the irradations from the detailed description of the physical interactions at the nanoscale to the prediction of the number of DNA damage, their complexity and their distribution in space.During my thesis, I developed micro-dosimetric models based on the Geant4-DNA modeling toolkit in two cases. The first concerns the simulation of the energy distribution deposited in a cell nucleus and the calculation of the number of different types of DNA damage (single and double strand breaks) at the nanometric and micrometric scales, for different types and numbers of delivered particles. These simulations are compared with experimental measurements of the kinetics of GFP-labeled (Green Fluorescent Protein) DNA repair proteins in human cells. The second is the dosimetry of irradiation of a multicellular organism to study the genetic instability in a living organism during development (C. elegans). I simulated the distribution of the energy deposited in different compartments of a realistic 3D model of a C. elegans embryo following proton irradiations. Finally, and in parallel with these two studies, I developed a protocol to characterize the AIFIRA microbeam using fluorescent nuclear track detector (FNTD) for proton and alpha particle irradiations. This type of detector makes it possible to visualize in 3D the incident particle tracks with a resolution of about 200 nm and to examine the quality of the cellular irradiations carried out by the CPM.

Les possibilités de combinaison thérapeutiques offertes par les nanoparticules ont ouvert un nouveau champ d’investigation pour la recherche sur le cancer. Dans ce projet de recherche, des nanoparticules dérivées de la protéine de capside du virus de la mosaïque du tabac (TMV) ont été utilisées afin de transporter différents peptides thérapeutiques ciblant le récepteur neuropiline-1. Cette stratégie a permis de solubiliser un peptide fortement hydrophobe ayant préalablement démontré son efficacité anticancéreuse sur des lignées de cancer du sein humain et de glioblastome. Les résultats obtenus ont également permis de démontrer la possibilité de combiner différents peptides thérapeutiques via l’auto-assemblage de la protéine de capside du TMV. / Nanoparticles play an ever increase role in carrying therapeutic compounds in the cancer field. In this research project, the coat protein of Tobacco mosaic virus (TMV) was used as nanocarrier to solubilize a hydrophobic peptide interfering with the transmembrane domain of neuropilin-1. The nanoparticles created have conserved the antiangiogenic and antimigratory effect of the therapeutic peptide. This strategy was also used to create nanoparticles carrying a peptide targeting the ectodomain of neuropilin-1. The two types of nanoparticles were then assembled through auto-assembling ability of the coat protein. These nanoparticles also exhibit antiangiogenic ability thus, confirming the validity of this approach to combine therapeutic peptides.

La lutte contre le cancer est certainement l’un des défis majeurs de ce 21ème siècle. Les résistances qui émergent contre les agents de thérapie ciblée présentent un aspect particulièrement épineux de cette problématique. La thèse présentée ici s’inscrit dans ce cadre. Elle vise à développer des molécules bioactives pouvant circonvenir les résistances apparues contre les traitements de certaines hémopathies malignes : la leucémie myéloïde chronique (LMC) et le syndrome myélodysplasique (SMD). Après avoir mis au point une méthodologie de synthèse monotope permettant de transformer un azoture en un 5-alcynyl-1,2,3-triazole, nous avons synthétisé deux séries de produits : nucléosidique et non nucléosidique. Pour chacune de ces séries, des relations structure-activité ont été établies. Après plusieurs cycles d’optimisation, trois composés lead très efficaces contre des lignées cellulaires résistantes de LMC et SMD, ont été sélectionnés. De surcroît, leur mode d’action s’est révélé très intéressant : il repose (partiellement ou entièrement, suivant le composé) sur un processus cellulaire qui connaît un véritable regain d’intérêt, à savoir l’autophagie. Une évaluation in vivo a été réalisée et a permis de valider l’activité prometteuse de notre composé lead nucléosidique. Par ailleurs, des études visant à déterminer la localisation intracellulaire et les cibles moléculaires de nos produits sont actuellement en cours / The fight against cancer is certainly one of the biggest challenges of the 21st century. Resistance that comes up against targeted therapy agents presents a particularly important aspect of this issue. The thesis presented here takes part within that framework. It aims at developing bioactive molecules able to circumvent resistance that have emerged against the treatment of certain hematological malignancies: chronic myeloid leukemia (CML) and myelodysplastic syndrome (MDS). Having developed a one-pot synthesis methodology that converts azides into 5-alkynyl-1,2,3-triazole, we synthesized two series of products: nucleosidic and non-nucleosidic. For each of these series, structure-activity relationships have been established. After running several cycles of optimization, three lead compounds particularly active on resistant cell lines of CML and MDS were selected. Further, their mode of action proved to be very interesting. It is based (partially or fully, depending on the compound) on a cellular process, which is experiencing a real renewed interest, the autophagy. An in vivo evaluation confirmed the promising activity of our nucleosidic lead compound. Moreover, studies aiming at determining the intracellular localization and molecular targets of our products are currently in progress

Prostate cancer is one of the most abundant types of cancer among men and the demand for a specific treatment is very high. In this thesis, I have focused on using Glutamate Carboxypepti- dase II (GCPII), as a target for a proof-of-principle delivery system. GCPII is a transmembrane protein that internalizes after a binding of a ligand and is overexpressed in prostate cancer. Virus-like particles from Murine polyomavirus (VLPs) are a suitable nanocarrier for the delivery of imaging agents and drugs. Here I describe modifying these VLPs with inhibitors of GCPII and fluorescent dyes and characterize their binding to GCPII on surface plasmon resonance and to cells expressing GCPII on confocal microscopy. VLPs carrying a GCPII inhibitor show specific binding to GCPII on surface plasmon reso- nance, however they bind non-specifically to cells that don't express GCPII. Several approaches have been tried to avoid that. The substitution of BC loop on the exterior surface of VLPs that is partially responsible for the binding of sialic acid did not seem to affect specificity on cells. Another approach tested was coating of the wild-type VLPs with large polymer carrying a flu- orescent label and a GCPII inhibitor. After the conjugation of the polymer to the VLP, specific binding and internalization in GCPII-positive...

TITLE: Support for the development of reading literacy and the practice of Czech schools AUTHOR: Mgr. Pavlína Nešněrová DEPARTMENT: Department of Primary Education PedF UK Prague SUPERVISOR: Prof. PaedDr. Radka Wildová, CSc. ABSTRACT: The subject of this dissertation thesis is the issue of reading literacy of pupils of primary education. The aim of the thesis is to analyze the quality of reading literacy of pupils of the first grade of elementary school through specifically focused questions and tasks and to elaborate a practical methodical manual for teachers on this topic. The research itself was realized through a didactic test aiming to determine the reading literacy status of pupils of 3rd, 4th and 5th year of elementary school through four groups of questions and tasks in terms of the ability to obtain and process information in various types of texts, texts. Based on the results, conclusions were formulated and discussion topics presented. The research findings were applied in a practical manual, which is part of the work, and which is intended to inspire teachers to work with literary texts in lessons. KEYWORDS: Reading literacy, primary education, targeted tasks, skills and competences, text information.

Glycosidases (EC 3.2.1.) alias glycoside hydrolases are enzymes that catalyze the cleavage of a glycosidic bond between two carbohydrates or between a carbohydrate and an aglycone. Under suitable conditions (especially reduction of water activity in the reaction mixture), these enzymes are also able to synthesize a glycosidic bond. By targeted mutagenesis of the catalytic centre of the enzymes, it is possible to suppress or completely abolish their hydrolytic activity. Enzyme synthesis using glycosidases makes it possible to prepare bioactive galactosides, for example galectin ligands. The present work deals mainly with β-galactosidase from Bacillus circulans, its recombinant expression and mutagenesis. In the first part of the work, the commercially prepared plasmid of -galactosidase from B. circulans isoform A that I designed was used for recombinant expression in E. coli. It was necessary to optimize the conditions of the enzyme production. As it is a large protein (189 kDa), the expression vector pCOLD II and cold production at 15 ř C were used. The enzyme is specific for the formation of the β-1,4 glycosidic bond and has been used to synthesize complex tri- and tetrasaccharide ligands that cannot be prepared with a crude commercial preparation containing undesirable enzyme activities....

Parental support programs aimed at foreign-born parents have increased in interest in Swedish municipalities. Research on these types of parental support programs is therefore important in order to ensure that the support given have the desired effects and that the programs have value for the individuals to whom the support is directed. The aim of this thesis was to identify the goals of a parental support program offered to foreign-born fathers as well as to investigate and analyse how participants in the parental support program experience and understand their participation in the program in relation to these goals. The empirical material was collected through qualitative textual analysis of background documents to the parental support program as well as through qualitative interviews with three participants in the program. The empirical material was analysed through theories of empowerment and gender. The result showed both similarities and differences between the participants experiences of the parental support program and the background documents that formed the basis of the program. The main result showed that the goal of the parental support was to fundamentally change the fathers’ way of being, by promoting gender equality and by inspiring fathers to take greater responsibility for their children. However, the interviewed fathers held a pragmatic attitude towards the parental support program, by using the information as a tool to maneuver in Swedish society without necessarily having to fundamentally change.

Despite the central role of IFNγ in vitiligo pathogenesis, systemic IFNγ neutralization is an impractical treatment option due to strong immunosuppression. However, most vitiligo patients present with less than 20% affected body surface area, which provides an opportunity for localized treatments that avoid systemic side effects. After identifying keratinocytes as key cells that amplify IFNγ signaling during vitiligo, I hypothesized that tethering an IFNγ neutralizing antibody to keratinocytes would limit anti-IFNγ effects to the treated skin for the localized treatment. To that end, I developed a bispecific antibody (BsAb) capable of blocking IFNγ signaling while binding to desmoglein expressed by keratinocytes. I characterized the effect of the BsAb in vitro, ex vivo, and in a mouse model of vitiligo. SPECT/CT biodistribution and serum assays after local footpad injection revealed that the BsAb had improved skin retention, faster elimination from the blood, and less systemic IFNγ inhibition than the non-tethered version. Furthermore, the BsAb conferred localized protection almost exclusively to the treated footpad during vitiligo that was not possible by local injection of the non-tethered anti-IFNγ antibody. Thus, keratinocyte-tethering proved effective while significantly diminishing off-tissue effects of IFNγ blockade, offering a new treatment strategy for localized skin diseases, including vitiligo.

The use of drones in conflicts is under development and is increasing rapidly. The first real drone warfare was seen in Pakistan in the War on Terrorism in 2004. Between officials and scientists, there are divided opinions about whether the drone strikes can increase terrorism or cause the collapse of organizations. The purpose of this thesis is to examine how the effectiveness of drones, in terms of precision and lethality, can be seen to escalate or de-escalate a conflict. The aim is to explore whether two existing theories about air power can explain the case of killing the Iranian general, Qasem Soleimani, in 2020. As a qualitative case study, the approach results in favor of Pape´s theory over Warden´s. Pape says that decapitation is more likely to escalate a conflict, which confirms this case. Other explanations in his theory are the absence of unexpected political effects and the overthrow of government. Though Wardens theory shows some aspects of de-escalation, the conclusion is that Soleimani was not the center of gravity that was meant to cause system collapse. Further research is required since the findings cannot identify any escalating or de-escalating effects within the Quds Force.