Cette thèse porte sur la méthode de marquage métabolique de glycanes. Elle consiste à utiliser des monosaccharides modifiés pouvant être métaboliquement introduits sur la membrane externe des cellules. Deux cibles ont été choisies, les bactéries Legionella pneumophila, et des cellules eucaryotes (lignée cellulaire du cancer de la prostate, PC3).Différents analogues saccharidiques ont été synthétisés pour les deux cibles portant différents rapporteurs chimiques, notamment des fonctions azoture, alcyne terminal, alcène terminal, cyclopropène et cétone. Une voie de synthèse commune a été développée mettant en jeu des dérivés d’esters de N-hydrosuccinimide et des dérivés saccharidiques amino, tels que la D-mannosamine, la D-galactosamine et le L-fucose pour les cellules eucaryotes et des analogues d’un précurseur de l’acide légionaminique pour les bactéries Legionella pneumophila.Des essais de marquages métaboliques de glycanes ont été effectués sur Legionella pneumophila. Pour cela, différents dérivés saccharidiques ont été incorporés par les bactéries, puis la révélation des rapporteurs chimiques avec des groupements complémentaires a été évaluée (azoture-cyclooctyne, alcène-tétrazine, cétone-hydrazide/alkoxyamine). La détection a été réalisée soit directement (fluorophore sur le partenaire), soit indirectement (reconnaissance d’un groupement biotine par une streptavidine portant un fluorophore). Les bactéries ont été ensuite observées par microscopie photonique et les observations ont mis en évidence un marquage membranaire.Pour les cellules eucaryotes (PC3), des essais de marquage métabolique ont été effectués afin de vérifier l’incorporation des monosaccharides modifiés via une détection par fluorescence. Une série d’outils portant les fonctions complémentaires aux rapporteurs chimiques, notamment un dérivé de cyclooctyne (TMDIBO) et un dérivé de tétrazine ont été synthétisés. Ils ont été couplés à des dérivés de biotine afin d’obtenir des outils pour la microscopie photonique et à des ARMs (Antibody Recruiting Molecules), tel que le 2,4-dinitrophényle et le rhamnose, en vue d’une potentielle approche thérapeutique. Celle-ci consiste à combiner l’incorporation de monosaccharides modifiés et leur réaction avec un partenaire portant des ARMs, afin de recouvrir la surface des cellules par ces motifs. Les ARMs en présence de sérum humain vont ensuite activer le complément conduisant à la lyse des cellules marquées. Des tests de marquage métabolique ont été réalisés avec les outils couplés aux ARMs et une détection par fluorescence a permis de vérifier la présence des ARMs à la surface des cellules. Des premiers essais ont été effectués avec du sérum humain et des optimisations sont à réaliser.Le marquage métabolique de glycanes est une méthode efficace afin de détecter par fluorescence les bactéries Legionella pneumophila à l’aide d’analogues d’un précurseur de l’acide légionaminique et les cellules eucaryotes PC3 via des dérivés d’autres monosaccharidiques. Le marquage obtenu dans les deux cas est membranaire. Cette méthode permet éventuellement de combiner des aspects d’imagerie ou de diagnostic, avec différentes approches thérapeutiques. / This PhD work focuses on glycans metabolic labeling. This method uses a modified monosaccharides bearing a chemical reporter. The unnatural monosaccharide is metabolically incorporated into glycans. Two targets were selected, Legionella pneumophila bacteria and prostate cancer cells (PC3).Various saccharidic analogs were synthesized carrying several chemical reporters, like azide, terminal alkyne, terminal alkene, cyclopropene and ketone functions. A common synthetic strategy was developed using N-hydrosuccinimide ester derivatives and amino monosaccharides, such as D-mannosamine, D-galactosamine and L-fucose for eukaryotic cells and analogs of a legionaminic acid precursor for Legionella pneumophila bacteria.Differents metabolic labeling of glycans were carried out on Legionella pneumophila. Various sugar derivatives were incorporated by bacteria, then the reporter group was reacted selectively and covalently with a complementary function (azide-cyclooctyne, alkene-tetrazine, ketone-hydrazide/alkoxyamine). Bacteria were visualized with an imaging probes by light microscopy (directly: partner bearing a fluorophore, indirectly: recognition of a biotin group by a fluorescent streptavidin). The results highlighted outer membrane labeling.For eukaryotic cells (PC3), metabolic oligosaccharides engineering has been accomplished to check chemical reporter analogs incorporation via detection with fluorescent probes. A series of tools carrying functions complementary to the chemical reporters was synthesized, such as cyclooctyne derivatives (TMDIBO) and tetrazine derivatives. These derivatives were combined with biotin groups for detecting tools or with Antibody Recruiting Molecules (ARMs), such as 2,4-dinitrophenyl and L-rhamnose, for potential therapeutic approaches. The concept focuses on the combination of metabolic glycan labeling and the activation of human serum complement by ARMs to kill selectively labeled cells. First, ARMs labeled cells was checked by recognition of fluorescent anti-ARMs antibodies. Then, metabolic glycan labeling and human serum complement activation has been evaluated but the complement activation protocols still need to be optimized.Metabolic labeling of glycans is an effective method to detect, by fluorescence, Legionella pneumophila bacteria using analogs of a legionaminic acid precursor and eukaryotic cells (PC3) with monosaccharide derivatives. The labeling was observed on the membrane of the two targets. This method can potentially combine imaging or diagnostic aspects with various therapeutic approaches.

Transplantation of pancreatic islets (PIs) represents an alternative treatment for type 1 diabetes mellitus. Post-transplant monitoring of islets by a reliable imaging method may contribute to the improvement of the transplantation outcome. In this thesis, novel visualization approaches for PIs were tested using magnetic resonance (MR) and optical imaging on phantoms and experimental animals, including Chemical Exchange Saturation Transfer (CEST) MR, fluorine (19 F) MR, bioluminescence and fluorescence imaging. MR imaging based on frequency-selective method CEST was performed on islets labeled with Eu-/Yb-based chelates. Labeled islets possessed low MR signal in phantoms, what would have been unsatisfactory for in vivo applications. Moreover, viability and function of labeled islets was impaired reflecting limited applicability of these agents for islet labeling and visualization. Genetically modified bioluminescent islets showed suitable properties for longitudinal tracking of their post-transplant fate at an artificial transplant site - subcutaneously implanted polymeric scaffolds. Using multimodal imaging (MR and bioluminescence), the optimal timing for transplantation of islets into the scaffolds was assessed in diabetic rats. Islets transplanted into scaffolds using the optimized timing scheme...

Sequence labeling is a type of machine learning problem that involves as- signing a label to each sequence member. Deep learning has shown good per- formance for this problem. However, one disadvantage of this approach is its requirement of having a large amount of labeled data. Semi-supervised learning mitigates this problem by using cheaper unlabeled data together with labeled data. Currently, usage of semi-supervised deep learning for sequence labeling is limited. Therefore, the focus of this thesis is on the application of semi-super- vised deep learning in sequence labeling. Existing semi-supervised deep learning approaches are examined, and approaches for sequence labeling are proposed. The proposed approaches were implemented and experimentally evaluated on named-entity recognition and part-of-speech tagging tasks.

La presente investigación tuvo como objetivo principal determinar los factores que influyeron en las exportaciones de la leche evaporada con partida arancelaria 0402911000 hacia el mercado de los Estados Unidos durante el periodo 2008-2018. La metodología que se usó para el estudio fue de tipo descriptiva, con diseño no experimental, con un corte longitudinal y con un enfoque mixto al intervenir variables cualitativas y cuantitativas, tales como las medidas sanitarias y fitosanitarias, el valor FOB y la producción. El principal resultado en el enfoque cualitativo fue el grado de influencia de las exigentes medidas sanitarias que impactó en el desempeño de las exportaciones al mercado estadounidense, debido a que, las normas norteamericanas difieren de las normas internacionales. Por otro lado, en el enfoque cuantitativo, el hallazgo en la competitividad del valor FOB ha favorecido las exportaciones hacia el mercado estadounidense. Sin embargo, en la producción, pese a que se cuenta con la materia prima necesaria para abastecer mercados internacionales, la falta de apoyo por parte del Estado para orientar a los productores es la causa de fondo del porque no se ha llegado a la competitividad esperada. La población de estudio está conformada por 16 gerentes y jefes de las empresas Gloria y Nestlé; En donde se aplicó entrevistas semiestructuradas a expertos como instrumento de recolección de datos. El estudio pretende responder si las normas de etiquetado son una barrera al comercio, y si la competitividad del valor FOB y la producción contribuye a las exportaciones. / The main objective of this investigation was to determine the factors that influenced the evaporated milk exports tariff heading 0402911000 to the United States market during the period 2008-2018. The methodology used for the study was descriptive, with a non-experimental design, with a longitudinal section and with a mixed approach, involving qualitative and quantitative variables, such as sanitary and phytosanitary measures, FOB value production. The main result in the qualitative approach was the degree of influence of the demanding sanitary measures that had and impact of the performance of exports to the US market, because the North American standards differ from the international standards. On the other hand, in the quantitative approach, the finding in the competitiveness of the FOB value has favored exports to the US market. However, in production, despite having the raw material necessary to supply international markets, the lack of support from the State to guide producers is the root cause of why the expected competitiveness has not been reached. The study population is made up of 16 managers and heads of the companies Gloria and Nestle; Where semi-structure interviews with experts were applied as an instrument for data collection. The study aims to answer whether labeling rules are a barrier to trade, and whether the competitiveness of FOB value and production contributes to exports. / Tesis

Para el año 2018, ya se podían identificar -en países como Chile y Ecuador- resultados exitosos relacionados a la implementación de leyes relacionadas al etiquetado frontal. El fin único de la creación de estas regulaciones era reducir los porcentajes de muertes por enfermedades alimenticias y mejorar la alimentación de los ciudadanos en cada uno de esos países, y lo estaban logrando. Para replicar los buenos resultados, en el año 2018, el Gobierno Peruano decidió aprobar una ley que regule un sistema de etiquetado frontal, tomando como referencia a los octógonos chilenos. Por supuesto, ante el cambio, las empresas y los consumidores reaccionaron de diversas maneras. Este estudio se centra en los consumidores. Pasado un año de la aprobación de la ley, ¿Cuál será su percepción y comportamiento con respecto a los octógonos? El presente estudio tiene alcance correlacional y diseño no experimental, e investiga la percepción y comportamiento de los consumidores de galletas dulces de 18 a 55 años de Tambo del Parque San José de Jesús María con respecto a la implementación de octógonos. Esta categoría –galletas- fue elegida debido a que, según estudios, podría verse más afectada que otras. Paralelamente, este estudio busca conocer si el género y, edad o NSE influyen en la percepción y comportamiento de los mismos consumidores. Muestra: 399 consumidores de galletas dulces de 18 a 55 años de Tambo del Parque San José de Jesús María Resultados: Los consumidores de galletas tuvieron una buena aceptación al etiquetado nutricional, lo toman en cuenta al comprar y consumir la categoría. Esto no varía según género ni por nivel socioeconómico, pero si por rango de edad. / By 2018, successful results related to the implementation of l front-of-package labeling systems could already be identified in countries such as Chile and Ecuador. The sole purpose of creating these regulations was to reduce the percentages of deaths from foodborne illness and improve healthy nutrition of citizens in each of these countries, and they were achieving it. To replicate the good results, in 2018, Peruvian government decided to approve a law that regulates a front-of-package labeling system, taking Chilean octagons as a reference. For that reason, in face of change, businesses and consumers reacted in several ways. This study focuses on consumers. One year after the approval of the law, what would be their perception regarding octagons? The present study is descriptive (non-probabilistic) and investigates the perception of consumers of sweet biscuits from 18 to 55 years of age in Tambo, Parque San Jose from Jesus Maria, respect to the implementation of octagons. This category -biscuits- was chosen because, according to studies, it could be more affected than others. At the same time, this study seeks to know whether gender, age or NSE influence the perception of the same consumers. Sample: 399 consumers of sweet biscuits from 18 to 55 years of age in Tambo, Parque San Jose from Jesus Maria Results: Biscuits consumers had a good acceptance of nutritional labeling; they take them in count when buying and consuming the category. This doesn’t vary according to gender or socioeconomic level, but it does by age range. / Tesis

Med kritisk diskursanalytisk metod har denna studie analyserat hur två svenska tidningar framställer hot och våld av elever mot lärare. Studien ämnar ta reda på vilket sätt hot och våld används som ett begrepp, vilken innebörd som skapas och vilka diskurser om hot och våld som dominerar i tidningarna. Materialet som analyserades bestod av 14 texter, både debattartiklar och nyhetsartiklar, publicerade i Svenska Dagbladet och Aftonbladet under perioden 2014–2019. Faircloughs tredimensionella modell användes för den kritiska diskursanalysen vilket resulterade i tre dominerande diskurser; hot och våld som socialt begrepp, hot och våld är av kriminell karaktär och stökiga elever i utanförskap. En underliggande diskurs som uppmärksammats är elevernas röst hörs inte. Utifrån Faircloughs teori om kritisk diskursanalys och stämplingsteori diskuterades samtliga diskurser. Resultatet visar att inom diskurserna om hot och våld framställs en stereotypisk föreställning om elever i socialt utanförskap och i kriminalitet som riskerar att stämpla eleverna. Vidare kom studien fram till att tidningarnas användning av orden hot och våld används som ett samlingsbegrepp där få ifrågasätter dess innebörd. Det diskuteras även hur användningen av samlingsbegreppet medför en risk att begreppet tillämpas även i situationer som inte är kopplade till brottsliga handlingar. Socialarbetare som kommer i kontakt med människor som utför eller utsätts för hot och våld behöver vara uppmärksamma på hur detta framställs i media och vilka konsekvenser detta kan tänkas få i bemötande, bedömning och beslut inom socialt arbete. / With a critical discourse analytical method this study has analyzed how two Swedish newspapers portrays threat and violence by pupils against teachers. This study aims to find out how threats and violence as a concept is used, which definition of threats and violence that is produced and what discourses about threats and violence that dominates in the newspapers. The analyzed material consisted of 14 texts, including debate articles and news articles, published in Svenska Dagbladet and Aftonbladet between the years of 2014-2019. Fairclough’s three-dimensional model was used and resulted in three dominating discourses; threats and violence as a social concept, threats and violence is of criminal nature and careless pupils in exclusion. An underlying discourse was that the pupils’ voices were not heard. All discourses were discussed using Faircloughs theory on critical discourse analysis and labeling theory. The result showed that within the discourses of threats and violence there is a stereotypical conception about students in social exclusion and in crime that is at risk of labeling them. Furthermore, this study found that the newspapers use of the words threat and violence is used as a concept where few question its meaning. It is also discussed how the use of the concept leads to a greater risk of describing situations that are not associated with criminal actions. Social workers who encounter people who use or is subjected to violence need to be attentive on how it is portrayed in the media and what consequences it might have in encounters, assessment and decision-making within social work.

No description available.

3 Selectively substituted cyclodextrins for analytical and pharmaceutical applications Abstract This thesis is focused on the selective modification of cyclodextrins, and its primary aim is the preparation and characterization of mono- and persubstituted derivatives of cyclodextrins in a regioselective and straightforward manner. The work is divided into two main parts describing synthetic strategies and applications of modified cyclodextrins with one or several substituents, respectively. The first section deals with the introduction of a single chromophoric moiety on the cyclodextrin scaffold such as cinnamyl, rhodaminyl, fluoresceinyl and eosinyl groups. The complete set of monocinnamyl-α-cyclodextrin regioisomers has been prepared by direct alkylation, and the self-assembling properties of the corresponding regioisomers were thoroughly investigated by dynamic light scattering and NMR experiments. These investigations revealed that the different isomers (mono-6-O-, mono-2-O- and mono-3-O- cinnamyl-α-cyclodextrin) form distinct supramolecular species through intermolecular association. A fast method for the unambiguous identification of the pure regioisomers has also been developed based on a series of 2D NMR measurements. Xanthene-modified β-cyclodextrins, other representatives of monosubstituted...

No description available.

Les glycanes sont des biomolécules constituées d’un enchainement de monosaccharides liés entre eux par des liaisons glycosidiques. La nature et l’abondance des monosaccharides constituant la chaine glycanique ainsi que l’agencement des motifs de glycosylation diffèrent fortement en fonction de l’organisme d’origine. La biosynthèse et la dégradation de ces architectures polysaccharidiques sont finement régulées par des systèmes enzymatiques spécifiques et sont organisées au sein des divers compartiments cellulaires. Les glycanes interviennent dans divers processus biologiques : réserves énergétiques, repliement et stabilité protéique, reconnaissance cellulaire et adhésion à la matrice extracellulaire.Cette thèse porte sur la synthèse et l’utilisation de composés permettant le marquage métabolique des glycanes de cellules eucaryotes (lignée cellulaire du cancer de la prostate, PC-3) ou de bactéries didermes à mycomembranes (Corynebacterium glutamicum).Les composés synthétisés ou utilisés dans cette thèse sont des saccharides fonctionnalisés par un groupement bio-orthogonal (groupements azido, alcyne ou méthylcyclopropène) ou des outils de marquage porteurs simultanément d’un groupement bio-orthogonal complémentaire (groupements cyclooctyne ou tétrazine) et d’une étiquette pouvant être détectée par une macromolécule fonctionnalisée par des fluorophores (D-biotine/stréptavidine ou 2,4-DiNitroPhénol/ anticorps anti-DNP). La solubilité, en milieu aqueux, des outils de marquage est un facteur limitant leur utilisation. Une partie des travaux de cette thèse a consisté à développer des outils de marquage plus solubles en milieu aqueux, par l’ajout d’un motif -sulfo--alanine. Ce motif porte une fonction acide sulfonique qui est déprotonée, à pH physiologique, favorisant ainsi la solubilité en milieu aqueux.L’incorporation métabolique des saccharides fonctionnalisés ainsi que la capacité des outils à marquer les cellules (après incorporation métabolique) est évaluée, sur l’un ou l’autre des modèles biologiques, par analyse en cytométrie en flux ou par microscopie confocale. / Glycans are biomolecules made up of a chain of monosaccharides bound together by glycosidic bonds. The nature and abundance ofmonosaccharides forming the glycanic chain and the arrangement of glycosylation patterns differ greatly depending on the organism of origin. The biosynthesis and degradation of these polysaccharide architectures are finely regulated by specific enzymatic systems and are organized within the various cell compartments. Glycans are used in various biological processes: energy reserves, protein folding and stability, cell recognition and adhesion to the extracellular matrix.This thesis focuses on the synthesis and use of compounds allowing the metabolic labeling of glycans of eukaryotic cells (cell line of prostate cancer, PC-3) or of diderm bacteria with mycomembranes (Corynebacterium glutamicum).The compounds synthesized or used in this thesis are saccharides functionalized by a bio-orthogonal groups (azido, alkyne or methylcyclopropene groups) or labeling tools simultaneously carrying a bio-orthogonal complementary function (cyclooctyne or tetrazine groups) and a label that can be detected by a macromolecule functionalized by fluorophores (D-biotin/streptavidine or 2,4-DiNitroPhenol/ anti-DNP antibodies). The solubility of labeling tools in aqueous media is a limiting factor. Part of the work of this thesis has been to develop more water-soluble labeling tools by adding a -sulfo--alanine pattern. This pattern has a sulfonic acid function, which is deprotonated at physiological pH, thus promoting aqueous solubility.The metabolic uptake of functionalized saccharides and the ability of tools to label cells (after metabolic uptake) are evaluated in either biological model, by flow cytometry analysis or confocal microscopy.