La compréhension et l'exploitation des processus biologiques, tels que la symbiose fixatrice d'azote (plante-bactéries) et la symbiose endomycorhizienne à arbuscules (plante-champignons), représentent un intérêt agronomique et écologique majeur. Les lipo-chitooligosaccharides (LCO) jouent un rôle essentiel dans la mise en place de ces deux symbioses. Il est donc important de mieux comprendre les mécanismes induits par ces molécules signal, et notamment l’interaction avec leur récepteur dans les plantes-hôtes.Les synthèses chimiques de ces molécules sont souvent longues, difficiles et les rendements finaux sont très faibles. Il est intéressant de s'intéresser à l'obtention d'analogues biologiquement actifs dont la synthèse serait plus efficace. Différents travaux ont montré qu'un cycle triazole pouvait être un bon mime d'une unité saccharidique. De plus, des travaux de modélisation des interactions protéine-ligand ont permis de supposer que l'unité II d'un LCO-IV pourrait avoir un rôle moins important dans la liaison avec le récepteur. Le but du projet a donc été de réaliser la synthèse de ces analogues de LCO-IV dont l'unité II est remplacée par un lien triazole.La stratégie développée a nécessité la synthèse de deux briques moléculaires, un monosaccharide propargylé en position 4 et un disaccharide comportant un azoture en position anomère. Grâce à la mise au point des différentes étapes de synthèse, ces deux briques ont pu être obtenues avec de bons rendements. La cycloaddition [3+2] catalysée au cuivre a permis d'obtenir le triazole souhaité et les premiers analogues de tétrasaccharides de manière efficace. L'utilisation préalable de protections adéquates a permis d'introduire la diversité moléculaire souhaitée, une fonction sulfate sur la position 6 de l'unité réductrice et différentes chaînes carbonées lipophiles sur l'amine de l'unité non-réductrice. Quatre analogues de LCO ont été obtenus avec de très bons rendements. Les tests de compétitions, effectués sur ces analogues, n'ont montré aucune affinité pour le récepteur. Une approche synthétique de nouveaux analogues par une C-glycosylation a alors été proposée. / Understanding and exploitation of biological processes, such as symbiotic nitrogen fixation (plant-bacteria) and arbuscular endomycorrhizal symbiosis (plant-fungi), represent important agricultural, ecological and societal interest. The lipo-chitooligosaccharides (LCO) play an essential role in the implementation of these symbioses, it is then important to better understand the mechanisms induced by these signaling molecules, and especially the interaction with host-plant receptors.The chemical syntheses of these molecules are often long, difficult and the final yields are very low, so it could be interesting to obtain biological active analogs more efficiently. Various studies have already shown that a triazole unit could be a saccharide mimic. Furthermore, modelling studies of protein-ligand interactions showed that the monosaccharidic unit II of a LCO-IV could have a less important role in the interaction with the receptor. The project aimed to synthesize these LCO-IV analogs in which unit II was replaced by a triazole link.The strategy was directed towards the synthesis of two glycostructures, a monosaccharide propargylated at C-4 position and a disaccharide with an azide at anomeric position. Various synthetic steps were performed to have access to these two intermediates in good yields. Copper-catalyzed [3+2] cycloaddition gave efficiently the desired triazole unit of tetrasaccharidic analogs. The prior use of adequate protections finally allowed to introduce the desired molecular diversity, a sulphate function at C-6 position of the reducing unit and different lipophilic carbon chains on the amine of the non-reducing unit, and thus to obtain four LCO analogs with very good yields. Competition tests performed on these analogs have shown no affinity for the receptor. A synthetic approach of novel analogs using a C-glycosylation step was then proposed.

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No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação_Paulo Senna.pdf: 2545028 bytes, checksum: af78616e8637cde7118d817db8c6f8b0 (MD5) Previous issue date: 2017-02-23 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Libidibia ferrea is a medium-sized tree known as jucá, found in the North and Northeast of Brazil. The use of various parts of the tree is very common in folk medicine, especially in the distant regions of large urban centers. Jucá fruits are used in the form of tea or decoction to prevent or treat inflammation. The fruits were collected in the urban area of the City of Manaus / Amazonas. After grinding, 300 g of jucá powder were extracted by maceration with methanol for five days to obtain the crude extract. The analyzes in HPLC-MS were performed on crude metanolic extract of juca fruit peels and crude hydrolyzed extract. Some hydrolysable tannins, HHDP acid, DHHDP acid and chlorogenic acid were identified. The crude extract was subjected to a liquid-liquid partition, to give the phases in hexane, chloroform, ethyl acetate and hydroalcoholic phase. Tests of antioxidant activity (DPPH and ABTS) and anti-inflammatory (LOX) were performed in the phases obtained. The ethyl acetate phase was more effective in all the tests (IC50 DPPH = 3.32, IC50 ABTS = 4.12, IC50 LOX = 8.62), while the hexane phase presented the worst results among the fractions (IC50 DPPH = 59.92, IC50 ABTS = 211.37, IC50 LOX = 140.45). The AcOEt phase was subjected to column chromatography using Sephadex LH-20 and methanol as the mobile phase. The resulting AcS2 fraction was fractionated in Sephadex LH-20 again using methanol as the mobile phase, whose AcSS2 fraction was subjected to solid phase extraction using as the mobile phase H2O/ACN mixtures. All fractions were analyzed in analytical HPLC and fractions 6 and 7 were pooled and ellagic acid and methoxygeraniine were identified, the elucidation of the substances occurred by negative-mode EM-NMR and NMR (MeOD, 500 MHz). Finally, antioxidant activity assays were performed for ellagic acid (IC50 DPPH = 49.57, IC50 ABTS = 60.00) and methoxygeraniine (IC50 DPPH = 8.59, IC50 ABTS = 6.49). The isolation method present in this work can be used to obtain other substances with antioxidant and anti-inflammatory activity and also for the quantification of these substances. / Libidibia ferrea é uma árvore de médio porte conhecida como jucá, encontrada na região Norte e Nordeste do Brasil. O uso de diversas partes da árvore é muito comum na medicina popular, principalmente nas regiões longínquas dos grandes centros urbanos. Os frutos de jucá são utilizados na forma de chá ou decocto para prevenir ou tratar inflamações. Os frutos foram coletados na zona urbana da Cidade de Manaus/Amazonas. Após triturado, 300 g de pó de jucá foram extraídos por maceração em metanol por cinco dias obtendo-se o extrato bruto. As análises em CLAE-EM foram realizadas no extrato metanólico bruto das cascas do fruto de jucá e no extrato bruto hidrolisado, sendo identificados alguns taninos hidrolisáveis, ácido HHDP, ácido DHHDP e ácido clorogênico. O extrato bruto foi submetido a partição líquido-líquido, obtendo-se as fases em hexano, clorofórmio, acetato de etila e fase hidroalcoólica. Foram realizados testes de atividade antioxidante (DPPH e ABTS) e anti-inflamatório (LOX) nas fases obtidas. A fase em acetato de etila apresentou maior eficácia em todos os testes (IC50 DPPH = 3,32, IC50 ABTS= 4,12, IC50 LOX = 8,62), enquanto que a fase em hexano apresentou os piores resultados entre as frações (IC50 DPPH = 59,92, IC50 ABTS= 211,37, IC50 LOX = 140,45). A fase AcOEt foi submetida a cromatografia em coluna utilizando Sephadex LH-20 e metanol como fase móvel. A fração resultante AcS2 foi fracionada em Sephadex LH-20 novamente utilizando metanol como fase móvel, cuja fração AcSS2 foi submetida a extração em fase sólida utilizando como fase móvel misturas de H2O/ACN. Todas as frações foram analisadas em CLAE analítica e as frações 6 e 7 foram reunidas e foram identificados o ácido elágico e a metoxigeraniina, a elucidação das substâncias ocorreu por EM-APCI modo negativo e RMN (MeOD, 500 MHz). Por fim, ensaios de atividade antioxidante foram realizados para o ácido elágico (IC50 DPPH = 49,57, IC50 ABTS= 60,00) e para a metoxigeraniina (IC50 DPPH = 8,59, IC50 ABTS = 6,49). O método de isolamento presente neste trabalho pode ser utilizado para a obtenção de outras substâncias com atividade antioxidante e anti-inflamatória e também para a quantificação dessas substâncias.

The thesis comprises the synthesis and spectroscopic characterization of novel lipid derivatives mainly of the cancerostatic 5-fluorouridine (5-FU). The chemical structure of the lipid residues resembles naturally occurring compounds, namely acyclic terpenes, waxes, and large ring ketones. They are positioned either at N(3) or at the O-2’,3’ cis-glycolic moiety of the nucleoside. The introduction of the lipophilic residues was performed either by ketalization of the glyconic hydroxyls or by direct alkylation as well as by Mitsunobu reaction at N(3) of the aglycone. The resulting nucleolipids were further converted to 2-(cyanoethyl) phosphoramidites as building blocks for automated solid phase nucleic acid synthesis. The latters were used for the preparation of a series of lipo-oligonucleotides which were studied with respect to their immobilization within artificial lipid bilayers and compared concerning immobilization rate and stability. Moreover, selected 5-FU nucleolipids were fluorophore-labelled and tested with respect to their cancerostatic activity towards human colon carcinoma cells (HT-29). Additionally, O-2’,3’-functionalized 5-FU ketals were covalently bound either to soluble chitosanes of various molecular weight or to chitosane foils. The latters were studied towards their enzymatic degradability. Last, the reaction of lipophilic phosphoramidites with the blood volume expander hydroxyethyl starch yielding enzymatically hydrolysable phosphodiester bonds was investigated.

La poly(2-isopropyl-2-oxazoline) (PIPOZ) est un polymère thermosensible qui possède une température de solution critique inférieure (LCST) autour de 40 °C en solution aqueuse. Les travaux présentés s’intéressent aux propriétés en solution aqueuse et aux interfaces, de l’homopolymère PIPOZ, d’une PIPOZ fonctionnalisée avec un groupement lipidique (lipo-PIPOZ) et de copolymères à blocs à base de poly(éthylène glycol) et de PIPOZ. Si elle est régulièrement comparée à son isomère structurel le poly(N-isopropylacrylamide) (PNIPAM), les études sur les propriétés en solution de la PIPOZ sont cependant moins complètes que celles sur le PNIPAM. Le premier objectif des travaux présentés ici est de parfaire la connaissance du comportement en solution de la PIPOZ en présence d’additifs. Les effets de sels et de solvants hydromiscibles sur la solubilité de la PIPOZ ont été investigués par turbidimétrie et microcalorimétrie sur trois homopolymères de masses moléculaires différentes. Contrairement aux solutions de PNIPAM, l’ajout de méthanol à la solution de PIPOZ ne conduit pas au phénomène de cononsolvency où la solubilité du polymère diminue pour une certaine gamme de fractions volumiques de cosolvant. L’effet a néanmoins été observé dans le cas de système PIPOZ/Eau/THF. L’effet de sels sur la solubilité de la PIPOZ suit la série Hofmeister. La présence de sels chaotropes (NaI et NaSCN) en solution ont révélé un effet bien plus important sur la solubilité de la PIPOZ que pour son isomère. Les valeurs de point troubles de la solution de PIPOZ augmentent de plus de 30 °C pour une concentration en sel supérieure à 1 M. L’autre objectif de cette thèse est de synthétiser un système à base de PIPOZ capable de s’auto-assembler à l’interface air-eau afin de former des films interfaciaux par la technique Langmuir-Blodgett. A cette fin, un amorceur contenant un groupement lipidique (2 chaînes alkyles et un groupement phosphate) a été synthétisé et utilisé pour la polymérisation cationique par ouverture de cycle (CROP) du monomère 2-isopropyl-2-oxazoline conduisant à l’obtention d’un lipo-PIPOZ (Mn = 10 kg.mol-1). L’effet de deux sels (NaSCN et NaCl) sur les films interfaciaux a été étudié. Malgré leur effet opposé sur la solubilité de la PIPOZ en solution, ils conduisent tous les deux à l’expansion de la monocouche de lipo-PIPOZ. Transférés sur des substrats de mica, ces films ont été visualisés par microscopie à force atomique (AFM). La iv présence de sels dans la sous-phase lors de la formation de monocouches conduit à la formation d’agrégats d’épaisseur ~ 10 nm dont le diamètre augmente avec la concentration en sel. Enfin, le dernier objectif est de caractériser les propriétés en solutions de copolymères à blocs PIPOZ-b-PEG-b-PIPOZ. La polymérisation par CROP de la 2-isopropyl-2-oxazoline a été amorcée à partir d’un PEG (Mn = 2 kg.mol-1) bifonctionnel, Le polymère synthétisé (TrOH, Mn = 11 kg.mol-1) a ensuite subit une fonctionnalisation des extrémités de chaînes par des groupements octadécyles conduisant à l’obtention d’un copolymère à blocs téléchélique amphiphile et thermosensible (TrC18). Les propriétés des copolymères en solution aqueuse ont été étudiées par turbidimétrie, diffusion dynamique de la lumière (DLS), microcalorimétrie (DSC), microscopie électronique à transmission et spectroscopie à sonde fluorescente, FT-IR et AFM. Les deux copolymères sont thermosensibles et présentent des valeurs de points troubles de ~ 48 °C pour le copolymère TrOH et de ~ 38 °C pour le copolymère amphiphile. Ce dernier s’auto-assemble à température ambiante et forme, en solution aqueuse, des micelles de type fleurs de rayon hydrodynamique RH ~ 8 nm. L’effet prolongé de la température sur la cristallisation des blocs de PIPOZ a aussi été examinée. Les deux polymères cristallisent en solution aqueuse conduisant à la formation de fibres insolubles dans l’eau. Mots- / Poly(2-isopropyl-2-oxazoline) (PIPOZ) is a thermosensitive polymer whose lower critical solution temperature (LCST) in water is ~ 40 °C. This thesis focuses on the properties in aqueous solution and on interfaces of new poly(2-isopropyl-2-oxazoline) systems. PIPOZ is often compared to its structural isomer, the renowned poly(N-isopropylacrylamide) (PNIPAM). If PNIPAM has been the center of thermosensitive polymer research for the last three decades, it is PIPOZ which has recently been gaining interest. The first aim of the thesis is to improve on the knowledge on PIPOZ properties in aqueous solution in the presence of water-soluble additives. Effect of salts and cosolvents were investigated by turbidimetry and microcalorimetry (DSC) on PIPOZ homopolymers of different molecular weights. Effect of salts on PIPOZ solubility follows the Hofmeister series. Chaotropic anions (SCN-, I-) induce a large increase (up to 30 °C) of the cloud point temperature of PIPOZ solution which is 10 times larger than for PNIPAM. Adding methanol into PNIPAM aqueous solution leads to a decrease in solubility of the polymer. This phenomena is called cononsolvency. Unlike PNIPAM solutions, the addition of methanol in PIPOZ solution does not lead to a cononsolvency effect. Nevertheless, cononsolvency has been observed in the case of THF addition into PIPOZ aqueous solutions. The second aim of this work was to design and synthesize an amphiphilic PIPOZ able to anchor itself at the air-water interface and to form stable monolayer via the Langmuir-Blodgett technique. For that purpose, a lipidic initiator containing two alkyl chains and a phosphate group, was synthesized and used to initiate the cationic ring opening polymerization (CROP) of 2-isopropyl-2-oxazoline. The obtained amphiphilic (lipo-PIPOZ, Mn = 10 kg.mol-1) forms stable monolayers at the air-water interface. The presence of salt (NaCl or NaSCN) in the sub-phase during the compression of the films leads to expansion of the monolayer even if the salts have opposite effect on PIPOZ solubility in solution. The interfacial films were then transferred onto mica substrates and captured by atomic force microscopy (AFM). The salts induced the formation of aggregates (height ~ 10 nm) whose diameter depends on the salt and its concentration. At last, a block copolymer, TrOH, containing a central poly(ethylene glycol) (PEG) (Mn = 2 kg.mol-1) and two PIPOZ blocks was obtained by CROP of 2-isopropyl-2-oxazoline initiated vi by a bi-functionnal PEG. The total molecular weight was Mn ~ 11 kg.mol-1. Hydrophobic chain ends modification has been performed onto TrOH to bring amphiphilicity and to get a telechelic octadecyl-end capped block copolymer TrC18. The properties of these two block copolymers in water were characterized by dynamic light scattering (DLS), microcalorimetry (DSC), electronic transmission microscopy (TEM) and fluorescence spectroscopy, FT-IR and AFM. Cloud point temperature of copolymer solutions was found to be around 48 °C for TrOH and around 38°C for the amphiphilic analogue TrC18. The latter self-assembles at room temperature into flower micelles whose hydrodynamic radius is RH ~ 8 nm. Extended heating of both copolymer solutions leads to crystallization of PIPOZ block and insoluble fibers form in solution.

Les symbioses rhizobienne et mycorhizienne à arbuscules sont deux endosymbioses racinaires jouant des rôles importants dans le développement des plantes en améliorant leur nutrition minérale. Les lipo-chitooligosaccharides (LCOs), produits par les bacteries Rhizobia et les champignons mycorhiziens, sont essentiels pour l'établissement de la symbiose rhizobienne et stimulent la mycorhization. Chez la légumineuse Medicago truncatula, trois récepteurs-like kinase à motifs lysin (LysM), LYR3, NFP et LYK3 sont impliqués dans la perception des LCOs. Le travail présenté a eu pour objectif la caractérisation biochimique de ces récepteurs et leurs applications potentielles. Les orthologues de LYR3 de M. truncatula ont été clonés et se sont tous révélés, à l'exception de celui du lupin, capables d'établir une interaction d'affinité élevée avec les LCOs mais pas avec les chitooligosaccharides de structure apparentée. Afin de mieux comprendre les bases moléculaires de la reconnaissance des LCOs, des échanges de domaine entre les protéines LYR3 de lupin et de Medicago ont été effectués et ont révélé l'importance du troisième domaine LysM dans l'interaction. L'exploitation des capacités de reconnaissance des LCOs par LYR3 à des fins biotechnologiques a été évaluée à l'aide de récepteurs chimériques constitués du domaine extracellulaire de LYR3 et du domaine kinase des récepteurs immunitaires AtCERK1 et EFR. Il est apparu que LYR3 peut être utilisé pour élaborer des récepteurs chimériques mais leur mode d'activation reste à optimiser. Enfin l'étude des deux récepteurs symbiotiques NFP et LYK3 suggère qu'ils sont régulés par phosphorylation suite au traitement par les signaux symbiotiques. L'ensemble de ce travail apporte un éclairage nouveau sur les mécanismes de perception des LCOs et sur les modifications associées à leurs récepteurs qui en résultent. / Arbuscular mycorrhization and rhizobial nodulation are two major root endosymbioses which play important roles in plant development by improving their mineral nutrition. Produced by Rhizobia bacteria and mycorrhizal fungi, lipo-chitooligosaccharides (LCOs) were shown to be essential for the formation of the rhizobial symbiosis and to have stimulatory effects on mycorrhization. In the legume Medicago truncatula three lysin motif (LysM) receptor-like kinases LYR3, NFP and LYK3 have been shown to be involved in LCO perception. Here work is presented aimed at the biochemical characterization and application of these important receptor proteins. Cloned from several legume species orthologs of M. truncatula LYR3, except from lupin, were shown to bind LCOs with high affinity, but not structurally-related chitooligosaccharides (COs). Domain swaps between the lupin and Medicago proteins were used as a tool to decipher the molecular basis of LCO recognition and revealed the importance of the third LysM domain for LCO binding. The possibility of exploiting the LCO-binding capacity of LYR3 in biotechnology, through the composition of chimeric receptors, was investigated by combining together the extracellular domain of LYR3 protein with the kinases of Arabidopsis thaliana immune receptors, AtCERK1 and EFR. The results suggest that LYR3 could be used for constructing biologically active chimeric proteins whose mode of activation needs to be improved. Finally studies on the two LysM symbiotic receptors NFP and LYK3 suggest that they are regulated by changes in their phosphorylation after symbiotic treatments. Together this work brings light on the mechanisms underlying LCO perception and the modifications that receptors undergo after their treatment with LCO.

Le virus de l’hépatite C (HCV) infecte les hépatocytes. Il est remarquable par le fait q’ilperturbe fortement le metabolisme lipidique de l’hôte, conduisant à des dysfonctions majeures telles qu’une stéatose ou une résistance à l’insuline. In vivo, les virions sériques présentent une densité faible et variable reflétant leur association aux lipoprotéines de faible et de tres faible densité (LDL et VLDL). Ainsi, l'existence de lipo-viro-particules (LVP), contenant à la fois les constituants viraux et les apolipoprotéines B (apoB) et E (apoE) a été suggerée. Ces LVP joueraient un rôle important dans la persistance virale. Cependant, cette association entre HCV et apoB n'a pas été retrouvée in vitro avec les modeles cellulaires disponibles.Mes travaux de thèse se sont donc concentrés sur la mise en place d'un nouveau modèle deculture cellulaire capable de produire a la fois des VLDL et des particules virales HCV, permettantainsi d’étudier l’interaction entre les deux voies de synthèse. Dans un premier temps, lacaracterisation de la production de lipoproteines par differentes lignees d'hepatocytes a permis demontrer l'existence d'un défaut de secretion de VLDL en cellules Huh7.5, classiquement utiliséespour étudier HCV in vitro, alors que les cellules HepG2 et HepaRG sont capables de produire des VLDL physiologiques. Dans un second temps, des cellules HepG2 repliquant HCV de manière persistante ont été utilisées pour caractériser les particules virales produites. Etonnamment, a l'instar des cellules Huh7.5 et malgré leur capacité a produire des VLDL, les cellules HepG2 ne secreteraient pas de LVP mais des particules virales positives pour apoE et négatives pour apoB. / Hepatitis C virus (HCV) mainly infects hepatocytes. It is unique in its ability to impair host lipidmetabolism, leading to major liver dysfunctions as, for instance, hepatic steatosis or insulinresistance. In vivo, serum virions have a low and variable density, reflecting their association withlow- and very-low-density lipoproteins (LDL and VLDL). Hence, the existence of lipo-viro-particles(LVP), containing both viral components as well as apolipoprotein B (apoB) and E (apoE), has beensuggested. These LVPs could play an important role in viral persistence. However, this associationbetween HCV and apoB has not been observed in vitro, using the currently available cell culturemodels.Therefore, during my PhD, I have worked at setting up a new cell culture model, which wascapable of producing both VLDL and HCV particles, and therefore would enable the study of theinterplay between both synthesis pathways. First of all, we characterized lipoprotein production indifferent hepatocyte cell lines and confirmed that Huh7.5 cells, usually used to study HCV in vitro,were deficient for VLDL secretion, whereas two other cell lines, HepG2 and HepaRG, were able toproduce quasi-physiological VLDLs. Therefore, in a second time, we used HepG2 cells to replicate aHCV strain containing a selection gene and to characterize viral particle production. Surprisingly,VLDL-producing HepG2 cells were also unable to secrete LVPs, but rather secreted apoE-positive andapoB-negative viral particles, which were similar to ones Huh7.5 cells produced. This suggests thatthe ability to produce LVPs does not correlate with the ability to produce VLDL.

If you buy a new electric car today it will take on average about ten years for you to start saving money compared to just continue driving your old gasoline car. It will also take about 4 years until you start saving carbon dioxide emissions, both of this is because of new production costs and emissions. As the EU has banned producing new fossil fuel cars from 2035, it’s just a question of time before the power train in our cars will be electric. This rapid transition will lead to the older generation gasoline cars left by the road, with still usable chassis. This calls for a solution that uses this chassis but swaps out its power train for an electric one, reducing the initial cost and emission to drive electric. But is it that easy? In this mission, we take that technical question into our hands and convert a Swedish classic Volvo 340 from 1979 and give it an electric power train from 2022. We provide a detailed theory about the technology, a guide in choosing the right components, and the legal build requirements to pass the inspection. The conversion is done in an ordinary garage with standard tools and a welder. Using a small power-train with a maximum power of 30kW (40hp) and a battery size of 20kWh gave us a car with a maximum speed of 110km/h and a range of 150km. The project costed a total of 60 000 kronor, including the registration process. The technical legality and registration process went smoothly thanks to the Organisation SFRO (Sveriges fordonsbyggares riksorganisation) which takes care of the technical inspection and handles the paperwork. After one year and 10 000 km of driving and collecting data, we estimate that transitioning from gasoline to electric via a conversion compared to a new electric car reduces the economical investment return time from 10 to 2 years. And saves 8 tons of CO2. The battery used is secondhand. The second-hand market of electric car batteries, mostly from crashed or defective cars is growing and is estimated to be enough to convert the majority of old gasoline cars that are in good condition. The life length of second hand batteries in conversion is estimated to be equal to the rest of the chassis, due to the lower power requirements in conversion builds. We see that this idea has potential on a larger scale due to satisfying the criteria: Enough low complexity to do a conversion (if using common car models), lower cost and CO2 emissions then other options, supply for batteries exists, donor cars exist with chassis in good condition, market size is big enough and it’s legal to modify your car in Sweden and a few other countries. / Det är bara en tidsfråga innan drivlinan i våra bilar kommer att vara elektrisk, eftersom EU har lagt ett förbud mot att producera nya fossilbränslebilar från 2035. Men det är inte det enda trycket på förändring eftersom även bensinpriserna fortsätter att öka. En mycket hög efterfrågan förväntas på elbilar. Men för de flesta är det inte ett ekonomiskt alternativ att köpa en ny elbil, även om användandet är billigare tar det cirka 10 år innan du börjar se besparingar. Även produktionen av nya elbilar släpper ut koldioxid, motsvarande cirka 4 års körning av en bensinbil. Den snabba övergången kommer också att leda till att den äldre generationens bensinbilar skrotas, med fortfarande användbara chassi. Detta kallar på en lösning som använder dessa chassin men byter ut sin drivlinan till en elektrisk, vilket minskar den initiala kostnaden för att köra elbil samtidigt som man sparar in utsläppen från en ny produktion. Men är det verkligen så lätt? Vi tar vi frågan i våra händer och konverterar en svensk klassiker, en Volvo 340 från 1979 och ger den en drivlina från 2022. Vi ger en detaljerad teori om tekniken, en guide för att välja rätt komponenter och lagliga byggkrav för att klara besiktningen. Konverteringen görs i ett vanligt garage med standardverktyg och en svets. Ett år och tusen mil senare av körning och insamling av data uppskattar vi att en övergång från bensin till el via en konvertering jämfört med en ny elbil minskar den ekonomiska avkastningstiden för investeringen från 10 till 2 år. Och sparar 8 ton koldioxid. Batteriet som används är begagnat. Andrahandsmarknaden för elbilsbatterier, främst från kraschade eller defekta bilar, växer och beräknas räcka för att konvertera majoriteten av gamla bensinbilar som är i gott skick. Livslängden för begagnade batterier vid konvertering uppskattas vara lika med resten av chassit, på grund av de lägre effektkraven i konverteringsbyggen. Vi ser att denna lösning har potential i det större perspektivet, främst igenom utbudet och livslängd av batterier och chassin för konverteringar. Men också att kostnaden och kompexiteten är tillräkligt låg om de vanligaste modellerna används. Denna lösning är just nu bara tillgänglig i sverige och ett fåtal andra länder där det är lagigt att modifiera sina bilar.

This bachelor’s thesis aims to show an extensive overview of all the parts that build up an electric load driven longboard and see if a load controlled longboard can be seen as a safe, comfortable and convenient alternative to the more common remote controlled longboard. The thesis will also answer how weight can be measured on a longboard in the most effective way, what the most comfortable riding technique is and what a good motor-battery configuration to be able to travel at 30 km/h and 10 km would be. The longboard measures the weight distribution with load cells located between the deck and the trucks. An Arduino translates the input from the load cells to a certain speed and then sends it to an ODrive which controls a BLDC motor that is powered by two LiPo batteries. The results show that a load controlled longboard can very well be seen as a good alternative if right riding technique is used. The best technique is when the longboard accelerates when the rider tilts and keeps a constant speed when the rider stands straight. The best way to measure the weight is to fasten the trucks with hinges which lets the load cells register weight without anything interfering. Not all tests could be done because of Covid-19 but a measured top speed of 15 km/h with a high gear ratio is a promising result for the future when more suitable gear ratios will be tested to try to reach the goal of 30 km/h. / Det här kandidatexamensarbetet strävar efter att visa en omfattande överblick på alla delar som bygger upp en elektrisk lastdriven longboard och se om en laststyrd longboard kan ses som ett säkert, komfortabelt och behändigt alternativ till den vanligare radiostyrda longboarden via handkontroll. Det här arbetet kommer också svara på hur vikt kan mätas på en longboard på ett så effektivt sätt som möjligt, vad som är den mest bekväma åkstilen och vad är en bra motor-batteri konfiguration för att kunna åka i 30 km/h och nå 10 km skulle vara. Longboarden mäter viktfördelningen med lastceller som är placerade mellan brädan och truckarna. En Arduino omvandlar indatan från lastcellerna till en specifik hastighet som den sedan skickar till en ODrive som kontrollerar en borstlös likströmsmotor som i sin tur är driven av två LiPo batterier. Resultaten visar att en laststyrd longboard kan mycket väl ses som ett bra alternativ om rätt åkstil används. Den bästa stilen är att longboarden accelererar när åkaren lutar sig och håller en konstant hastighet när åkaren står rakt. Det bästa sättet att mäta vikt är att montera truckarna på gångjärn som låter lastcellerna mäta vikt utan att något stör. Alla tester kunde inte utföras på grund av Covid-19 men en uppmätt topphastighet på 15 km/h med en hög utväxling är ett lovande resultat för framtiden när lämpligare utväxlingar kommer testas för att försöka nå målet på 30 km/h.

Zusammenfassung Jörn Gieseler Stoffwechseluntersuchungen bei klinisch gesunden Kühen unter besonderer Berücksichtigung der wasserlöslichen und fettlöslichen Antioxidantien. Medizinische Tierklinik, Veterinärmedizinische Fakultät, Universität Leipzig. Eingereicht im Oktober 2010. 97 Seiten; 25 Abbildungen; 11 Tabellen; 351 Literaturangaben; Schlüsselwörter: Rind, Stoffwechsel, wasserlösliche Antioxidantien, lipidlösliche Antioxidantien. Die Gesundheit und Leistung von Milchkühen sind an einen stabilen Stoffwechsel gebunden. Abweichungen in der Nährstoffversorgung, im Intermediärstoffwechsel sowie seitens der Umweltbedingungen wirken sich belastend auf den Stoffwechsel und damit auf die Gesundheit des betroffenen Organismus aus. Mit gezielten Untersuchungen, z.B. Blutuntersuchungen, kann kontrolliert werden, ob der Stoffwechsel physiologisch oder durch Imbalancen belastet oder gar gestört ist. Ziel der Untersuchung war es, die Aktivität der Glutathionperoxidase sowie die Konzentrationen der wasser- und fettlöslichen Antioxidantien im Blut von klinisch gesunden Kühen zu ermitteln, in Stoffwechseluntersuchungen mit einzubeziehen sowie den Einfluss von Laktation und Jahreszeit auf die o.g. Parameter zu prüfen. Versuchsanordnung: Insgesamt wurden bei 85 SB/HF-Kühen (7990 kg fettkorrigierte Milch/Jahr) folgende zwei Gruppen analysiert: Gruppe 1: Zur Kontrolle des Laktationsverlaufes wurden 10 Kühe zum Zeitpunkt der 4. - 5. Woche ante partum (ap), 1. Woche ap, 1 - 2 Wochen post partum (pp), 4 Wochen pp und 8 - 12 Wochen pp untersucht. Gruppe 2: Im Verlaufe eines Jahres wurden im Abstand von 6 Wochen jeweils 10 gesunde Kühe, die sich alle in der 2. - 4. Woche post partum (pp) befanden, untersucht. Stall- und Außentemperaturen wurden dabei berücksichtigt. Die Tiere der beiden Gruppen wurden nach der klinischen Untersuchung weiter chemisch auf folgende Parameter getestet: Glutathionperoxidase (GPX), wasserlöslichen Antioxidantien (ACW), fettlösliche Antioxidantien (ACL), ß-Hydroxybutyrat (BHB), Cholesterol (Chol), Bilirubin (Bili), Glutamat-Dehydrogenase (GLDH), Aspartat Amino Transferase (AST), Creatinkinase (CK), Albumin (Alb), Harnstoff (Hast), Calcium (Ca), anorganisches Phosphat (Pi) und Chlorid (Cl). Durch eine Rationsberechnung wurde die Fütterung in die Untersuchung mit einbezogen. Ergebnisse: Die Ergebnisse der peripartalen Stoffwechseluntersuchungen zeigen einen Anstieg der wasserlöslichen Antioxidantien (ACW) bis zur 4. Woche pp (p < 0,05). Ab 8 Wochen pp sinken die Konzentrationen wieder ab. Im Gegensatz dazu zeigt die Glutathionperoxidase (GPX) ihre höchste Aktivität bei den Trockenstehern. Es folgt eine starke Abnahme der Aktivität bis zur 4. Woche pp (p < 0,05) und ein Anstieg ab der 8. Woche pp (p < 0,05). Die Korrelation zwischen der Konzentration der ACW und der Aktivität der GPX verhält sich signifikant negativ. Die höchsten Konzentrationen der ACL liegen im Zeitraum des Trockenstehens, die niedrigste Konzentration 1. Woche ap - 4. Woche pp (p < 0,05). Ab 8 - 12 Wochen pp steigen die Konzentrationen der ACL wieder an. Die Stoffwechselparameter Harnstoff, Bilirubin, Cholesterol und BHB unterliegen Schwankungen über den gesamten Laktationsverlauf. Die AST-Aktivität erreicht ihren Höchststand 1-2 Wochen pp und liegt nur in dieser Zeit außerhalb der Toleranzgrenze. Die Albumin- und Pi-Konzentrationen sowie die CK-Aktivitäten bleiben im Laktationsverlauf konstant. Die Cl-Konzentration liegt in der 1. - 12. Woche pp unterhalb der physiologischen Grenze. Die Ergebnisse der Stoffwechseluntersuchungen im Jahresverlauf zeigen einen kontinuierlichen Anstieg der ACW von Februar an, mit Höchstwerten im April und August (p < 0,05). Danach erfolgt ein kontinuierlicher Abfall der Werte bis zum Dezember. Die GPX zeigt eine generelle Verminderung ihrer Aktivität von Februar bis August (p < 0,05), um dann im Oktober wieder anzusteigen. Ihre Höchstwerte liegen im Januar und Dezember. Die Aktivität der GPX und die Konzentrationen der ACW korrelieren sowohl im Jahresverlauf als auch im Laktationsverlauf signifikant negativ. Die Konzentrationen der ACL unterliegen im Jahresverlauf Schwankungen. Dennoch korrelieren sie mit den Konzentrationen der GPX signifikant positiv. Die Harnstoff- und BHB-Konzentrationen sowie die Aktivität der CK liegen im gesamten Jahresverlauf innerhalb der physiologischen Grenzen. Die CK-Aktivität erreicht, zusammen mit der Albuminkonzentration, ihre Höchstwerte im Mai. Die Albuminkonzentrationen unterliegen mit dem Bilirubin im Jahresverlauf relativen Schwankungen. Das Bilirubin hat seine niedrigste Konzentration im Dezember und Januar bzw. seine höchsten Konzentrationen im Juli und August. Die Aktivität der AST zeigt einen gleichmäßigen Anstieg in den Sommermonaten. Ihre niedrigsten Aktivitäten liegen im Dezember und Januar. Die Cholesterolkonzentration sowie die Aktivität der AP sinken im Sommer ab. Cholesterol hat seine höchsten Konzentrationen im Dezember und Januar. Schlussfolgerung: Schwankungen von Stoffwechselparametern im Jahres- und Laktationsverlauf betreffen vor allem die Antioxidantien. Deshalb können sowohl die in dieser Arbeit untersuchten Antioxidantien, als auch die anderen Stoffwechselparameter zur Beobachtung und Diagnostik von Stoffwechselbelastungen rund um die Kalbung sowie im Jahresverlauf zur Herdenkontrolle herangezogen werden. Die Referenzbereiche betragen für die ACW 12 bis 142 µmol/l und für ACL 1 bis 45 µmol/l.

Submitted by PPG Pediatria e Sa?de da Crian?a (pediatria-pg@pucrs.br) on 2018-07-12T11:24:06Z No. of bitstreams: 1 Disserta??oLu SteffenVers?oFinal.pdf: 1086203 bytes, checksum: 9b8f206613da0e87460cd6110bcec5ac (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-07-17T11:24:05Z (GMT) No. of bitstreams: 1 Disserta??oLu SteffenVers?oFinal.pdf: 1086203 bytes, checksum: 9b8f206613da0e87460cd6110bcec5ac (MD5) / Made available in DSpace on 2018-07-17T11:32:31Z (GMT). No. of bitstreams: 1 Disserta??oLu SteffenVers?oFinal.pdf: 1086203 bytes, checksum: 9b8f206613da0e87460cd6110bcec5ac (MD5) Previous issue date: 2017-03-22 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Introduction: Cystic Fibrosis is a disease characterized by recurrent infections and chronic inflammation of the respiratory system that lead to irreversible pulmonary complications. Infections are mainly caused by Staphilococcus aureos(SA) and Pseudomonas aeruginosa (PA). Early diagnosis for identification of colonizing germs is an important challenge. Guidelines suggest the use of swab cultures of the oropharynx or sputum. However, studies on molecular testing, or alternative methods of collection are still unconclusive. The involvement of the upper airway (nasopharynx and paranasal sinuses) has been cited as the primary source of infection. The present study aims to describe and compare the most frequent findings and pathogens in the nasal tract in patients with cystic fibrosis and to correlate findings with markers of severity and progression of lung disease. Methods: This is a retrospective study, which included patients with a diagnosis of Cystic Fibrosis (CF) who are followed up at the Multidisciplinary Outpatient Clinic of the Pediatric Pulmonology Unit of the Hospital S?o Lucas (Pontifical Catholic University of Rio Grande do Sul - PUCRS). Patients who have performed otorhinolaryngological evaluation (ENT) in the last two years (2015-2016) were included. The video naso endoscopy and nasal swab collection were part of the ENT evaluation. Results: 48 patients with CF were included, of which 30 (62.5%) were male. The mean age was 12.15 years ?6.60, and the mean predicted forced expiratory volume in the first second (FEV1%) was 83.36 ?30.04. When evaluating the presence and characteristic of nasal secretion, only 9 patients (18.7%) presented purulent secretion. Twenty-six patients (54.2%) presented grade 1 tonsils and 12 (25%) grade 2 or 3 patients. Nasal swab bacteriology was positive in 26 (54.1%) patients, from which 22 presented Staphylococcus aureos, 2 Pseudomonas aeruginosa, 1 Pseudomonas cepacea and 1 Stenotrophomonas maltophila (SM). In 22 patients (45.8%) the result of the culture was negative. In this study, patients who presented positive colonization by the traditional method (oropharynx / sputum) had a statistically significant chance of being identified also by nasal cavity culture (p<0.001). However, the association was not perfect, and showed a low correlation for the detection of gram negative germs. Nasal polyps were observed in 9 participants. When polyp is used as a marker of disease in nasal endoscopy, a strong association is observed between the presence of polyps and lower Shwachman-Kulczycki clinical score (p <0.001). Conclusions: the results of the culture obtained by collection of the nasal cavity were similar to those found by the standardized collection methods as markers of colonization of the inferior airway. In addition, nasal swabs is characterized as a non-invasive technique and showed to be sensitive to the identification of relevant pathogens in CF, especially SA. In addition, the presence of the polyp in the nasal cavity was shown to be associated with prognostic markers as Shwachman-Kulczycki clinical score. / Introdu??o: a fibrose c?stica ? uma patologia caracterizada por infec??es recorrentes e inflama??o cr?nica do sistema respirat?rio que levam a complica??es pulmonares, por vezes, irrevers?veis. As infec??es s?o causadas, principalmente pelos microorganismos Staphilococcusaureos(SA) e Pseudomonas aeruginosa(PA). O diagn?stico precoce para identifica??o dos germes colonizadores ? ainda um desafio. Consensos sugerem o uso de culturas de swab da orofaringe ou escarro. No entanto, pesquisa por testes moleculares como op??o, ou formas alternativas de coleta ainda s?o inconclusivos. O comprometimento da via a?rea superior (nasofaringe e seios paranasais) tem sido citada como fonte prim?ria de infec??o. O presente estudo tem por objetivo descrever e comparar as altera??es e os pat?genos mais frequentes no trato nasal em pacientes com fibrose c?stica e correlacionar os achados com marcadores de gravidade e progress?o da doen?a pulmonar. M?todos: este ? um estudo retrospectivo, que incluiu pacientes com diagn?stico de Fibrose C?stica (FC) que s?o acompanhados no Ambulat?rio Multidisciplinar de FC do Servi?o de Pneumologia Pedi?trica do Hospital S?o Lucas da Pontif?cia Universidade Cat?lica do Rio Grande do Sul (PUCRS), e que tenham realizado avalia??o otorrinolarigol?gica (ORL) entre os anos de 2015 e 2016. A videonasoendoscopia e a coleta de material da fossa nasal com o uso do swab fizeram parte da avalia??o ORL. Os marcadores de gravidade e progress?o da doen?a foram: ?ndice de massa corporal (IMC), volume expirat?rio for?ado no primeiro segundo(VEF1%) e o escore cl?nico de Shwachman-Kulczycki(S-K). Resultados: foram inclu?dos 48 pacientes com FC, sendo 30 (62,5%) do g?nero masculino. A m?dia de idade foi 12,15 anos ? 6,60, e a m?dia do percentual do valor previsto de volume expirat?rio for?ado no primeiro segundo (VEF1%) foi de 83,36 ? 30,04. Ao avaliar a presen?a e caracter?stica da secre??o nasal, apenas 9 pacientes (18,7%) apresentavam secre??o purulenta. Com rela??o as tonsilas far?ngeas, 26 pacientes (54,2%) apresentavam tonsilas grau 1 e 12 pacientes (25%) grau 2 ou 3. A bacteriologia do swab nasal foi positiva em 26 (54,1%) pacientes, onde 22 apresentavam Staphylococcus aureos, 2 Pseudomonas aeruginosa, 1 Pseudomonas cepacea e 1 Stenotrophomonas maltophila(SM). Em 22 pacientes (45,8%) o resultado da cultura foi negativo. Neste estudo, os pacientes que apresentavam coloniza??o positiva pelo m?todo tradicional (orofaringe/escarro), tinham uma chance estatisticamente significativa de serem identificados tamb?m pela cultura da cavidade nasal (p<0,001). Por?m a associa??o n?o ? perfeita, e demonstrou baixa correla??o para detec??o de germes gram negativos. Foram observados p?lipos nasais em 9 participantes. Quando utilizada a presen?a de p?lipo como marcador de doen?a na endoscopia nasal, observa-se uma forte associa??o entre a presen?a de p?lipos e o redu??o dos valores no escore cl?nico de Shwachman-Kulczycki (p<0,001). Conclus?es: os resultados da cultura obtidos pela coleta da fossa nasal foram semelhantes aos encontrados pelos m?todos de coleta padronizados como marcadores de coloniza??o da via a?rea inferior. Al?m de caracterizar-se como uma t?cnica pouco invasiva, o swab nasal mostra-se sens?vel ? identifica??o de pat?genos relevantes na FC, especialmente SA. Al?m disso, a presen?a do p?lipo na cavidade nasal mostrou ser um dado associado a marcadores de progn?stico medido pelo escore cl?nico de Shwachman-Kulczycki.