Introduction: Le supplément d’oxygène et la nutrition parentérale (NP) sont les deux sources majeures de stress oxydant chez le nouveau-né. Lors de la détoxification des oxydants, le potentiel redox du glutathion s’oxyde. Notre hypothèse est que le supplément d’oxygène et la durée de la NP sont associés à un potentiel redox plus oxydé et à une augmentation de la sévérité de la dysplasie bronchopulmonaire (DBP). Patients et Méthodes: Une étude observationnelle prospective incluant des enfants de moins de 29 semaines d’âge gestationnel. Les concentrations sanguines de GSH et GSSG à jour 6-7 et à 36 semaines d’âge corrigé étaient mesurées par électrophorèse capillaire et le potentiel redox était calculé selon l’équation de Nernst. La sévérité de la DBP correspondait à la définition du NICHD. Résultats: Une FiO2≥ 25% au 7ième jour de vie ainsi que plus de 14 jours de NP sont significativement associés à un potentiel redox plus oxydé et à une DBP plus sévère. Ces relations sont indépendantes de l’âge de gestation et de la gravité de la maladie initiale. La corrélation entre le potentiel redox et la sévérité de la DBP n’est pas significative. La durée de la NP était responsable de 15% de la variation du potentiel redox ainsi que de 42% de la variation de la sévérité de la DPB. Conclusion: Ces résultats suggèrent que l’oxygène et la NP induisent un stress oxydant et que les stratégies visant une utilisation plus judicieuse de l’oxygène et de la NP devraient diminuer la sévérité de la DBP. / Introduction: oxygen supplementation and total parenteral solution (TPN) are two main clinical practices that sustain oxidative stress. Glutathione is a key molecule that detoxifies peroxides resulting in a more oxidized redox potential. We hypothesize that O2 supplementation and longer TPN duration are associated with both more oxidized redox potential and more severe bronchopulmonary dysplasia (BPD). Patients and methods: A prospective observational study including infants of less than 29 weeks gestational age. GSH and GSSG from whole blood sampled on day 6-7 and at 36 weeks of corrected age (CA) were measured by capillary electrophoresis and redox potential was calculated using Nernst equation. BPD was classified according to NICHD guidelines. Results: There was a significant association between FiO2 ≥ 25% on day 7 of life and TPN duration longer than 14 days and both more oxidized redox potential and more severe BPD. TPN duration explained both 15 % of total variation observed in redox potential and 42 % of total variation in BPD severity. These associations remained significant after adjustment for gestational age and illness severity. The relation between the severity of BPD and the redox potential in blood was not significant. The statistic power (1-β) to show an effect of redox potential on severity of BPD was 52%. Conclusion: Both redox potential of glutathione and BPD severity are both associated with early O2 supplement and TPN. Strategies targeting judicious use of O2 supplement and either decreasing the duration or using safer formulation of TPN are expected to help reducing BPD.

The focus of these studies has been toward the development of new synthetic methods and procedures for the synthesis of novel compounds with unique biological properties. This research has led to the development of two new synthetic strategies for the construction of two novel amphibian alkaloids. In addition, the efforts have led to the large-scale process for the preparation of a novel analgesic compound. The regioselective ring opening of lactones (δ-valerolactone and γ-butyrolactone) with aryllithium reagents is reported for the construction of a series of δ-hydroxyarylketones and γ-hydroxyarylketones. Both the R and S enantiomers of the amphibian alkaloid noranabasamine were prepared in >30% overall yield with 80% ee and 86% ee, respectively. An enantioselective iridium-catalyzed N-heterocyclization reaction with either (R)- or (S)-1-phenylethylamine and 1-(5-methoxypyridin-3-yl)-1, 5-pentanediol was employed to generate the 2-(pyridin-3-yl)-piperidine ring system in 69-72% yield. A cis-2, 5-disubstitued pyrrolidine building block derived from (-)-Cocaine•HCl was prepared. We utilized this compound as a chiral building block for the formal synthesis of (+)-gephyrotoxin. Using this pyrrolidine building block, Kishi's intermediate was obtained enantiospecifically in 15 steps and 9.4% overall yield. A large-scale process for the preparation of the analgesic compounds SCP-123 and its sodium salt, SCP-123ss•monohydrate has been developed. The process for the preparation of SCP-123 required three synthetic steps with no chromatography, while the process for the preparation of SCP-123ss required four synthetic steps and no chromatography. The overall yields for both SCP-123 and SCP-123ss were 47% and 46%, respectively, and both compounds were obtained in exceptionally high purity (>99%).

La centralisation de la préparation des médicaments injectables à la pharmacie permet d’assurer un haut niveau de qualité mais nécessite une stabilité du médicament dans le conditionnement choisi et une capacité de production suffisantes. Aseptic Technologies a développé un système de remplissage automatisé de conditionnement primaire utilisant un flacon en copolymères d’oléfines cycliques (COC) clos, stérile et prêt à l’emploi appelé « flacon Crystal® ». Ce flacon est rempli à travers le bouchon et est re-scellé par laser, réduisant ainsi le risque de contamination microbiologique.Nous avons évalué les risques liés à l’utilisation du flacon Crystal® :• Le risque de sorption est corrélé à la lipophilie de la substance conditionnée. Pendant 3 mois, nous avons évalué l’évolution de la concentration de 8 molécules cytotoxiques (de coefficients de partage octanol/eau différents) par CLHP/MS. Une méthodologie identique a permis le suivi des concentrations, pendant 24 heures de 3 molécules dérivées du platine par RMN.• Concernant le risque de perméation, nous avons évalué l’évolution des concentrations en phénol et métacrésol dans une solution à 1UI/mL d’insuline conditionnée dans des flacons Crystal® conservés droits et retournés pendant 50 jours par CLHP/UV. Deux formulations différentes de bouchons ont été testées.• Les solutions de pH extrêmes peuvent conduire à la présence de particules dans la solution injectable. L’impact sur la contamination particulaire du conditionnement de ces solutions dans les flacons Crystal® a été étudié et les particules caractérisées.• Les plastifiants peuvent migrer du conditionnement primaire vers la préparation parentérale. Après extraction par reflux à l’isopropanol sur le corps en COC et le bouchon en élastomère, les éventuels plastifiants ont été identifiés et dosés par RMN.Enfin, les stabilités de solutions de céfuroxime, midazolam et noradrénaline conditionnées en flacon Crystal® ont été déterminées et comparées à la seringue en polypropylène (PP).Les concentrations moyennes des 8 molécules cytotoxiques étaient comprises entre 90% et 110% à J62 sans corrélation avec le coefficient de partage. L’absence d’interaction a aussi été démontré pour le carboplatine et l’oxaliplatine par RMN. La précision de la quantification par RMN du cisplatine n’était pas suffisante.Il existe un risque de perméation du métacrésol lors du stockage à 25°C/60%RH pendant 50 jours (perte de 19,7% et de 20,3% pour les flacons Crystal® droits et retournés respectivement). La seconde formulation du bouchon permet de maîtriser le phénomène (perte de 2,6% à J50).Une contamination particulaire importante a été observée lors du conditionnement d’une solution basique et d’un contact avec le bouchon pendant 7 jours (6820 particules 3; 10µm dans un flacon Crystal® de 20mL) Les particules sont fibreuses et composées essentiellement de molécules inorganiques.Les analyses RMN mettent en évidence la présence de plastifiants dans le corps en COC et le bouchon à des concentrations largement en-dessous de la limite de 1000µg/g définie par la réglementation REACH.La solution de céfuroxime (1mL à 10mg/mL) était stable 365 jours à -20°C. Les solutions de midazolam (50mL à 1mg/mL) et de noradrénaline (20mL et 50mL à 0,2mg/mL ; 50mL à 0,5mg/mL) étaient stables 365 jours à 5°C. Des résultats similaires ont été obtenus lors de la conservation en seringue PP.Les flacons Crystal® ne sont pas sujets aux phénomènes de sorption. Il n’y a pas de risque de migration de plastifiants. Le risque de perméation du métacrésol est contrôlé par l’utilisation du nouveau bouchon. Le conditionnement de solutions basiques doit être surveillé. Les solutions de céfuroxime (à -20°C), de midazolam et de noradrénaline (à 5°C) sont stables 1 an. Par rapport à la seringue PP, le flacon Crystal® pourrait être préféré de par son système de remplissage aseptique automatisé ainsi que pour des considérations logistiques, en particulier le gain de place. / Centralized preparation of parenteral products in the pharmacy can ensure a higher quality but requires a stability of the drugs in the chosen primary packaging and a sufficient production capacity. Aseptic Technologies has developed an automated primary packaging filling system using a closed, sterile and ready-to-use cyclic olefin copolymer vial called ‘‘Crystal® vial’’. This vial is filled through the elastomeric septum and then immediately re-sealed by laser, thus reducing the risk of microbial contamination.We have evaluated risks linked to the use of Crystal® vials:- Sorption risk is correlated with substance lipophilicity. During 3 months, we have evaluated concentration evolution of 8 cytotoxic drugs (with different water/octanol partition coefficients) by HPLC/MS. Same methodology was used to measure concentration evolution of 3 platinum-derived drugs by NMR during 24 hours.- About the permeation risk, we have evaluated concentration evolution of phenol and metacresol in a 1UI/mL insulin solution during 50 days by HPLC/UV in Crystal® vials stored upright and upside down. Two stoppers formulations exist and were tested.- Extreme pH solutions can lead to the presence of particles in the parenteral product. Impact of these solutions in Crystal® vials on particles contamination was studied and particles characterized.- Plasticizers may migrate from the primary packaging to the parenteral products. After isopropanol reflux extraction on the COC body and thermoplastic elastomer stopper, potential plasticizers were identified and quantified using NMR.Finally, stabilities of ready-to-use cefuroxime, midazolam and noradrenaline solutions in Crystal® vials were determined and compared to polypropylene syringes.The 8 cytotoxic drugs mean concentrations were between 90% and 110% at day 62 with no correlation with partition coefficient. Lack of interactions was demonstrated too for the oxaliplatin and carboplatin solution by NMR. Accuracy of NMR cisplatin quantification was not sufficient.Studies performed on preservatives highlighted a metacresol permeation risk during storage at 25°C/60%RH for 50 days (loss of 19.7% and 20.3% respectively for Crystal® vials stored upright and upside down). Use of the second formulation stopper allowed a large decrease of permeation (loss of 2.6% at day 50).An important particles contamination was observed only with extrem alkaline pH and contact with the stopper (6820 particles 10µm in a 20mL Crystal® vials stored upside down during 7 days). Particles are fibrous and essentially composed by non-organic substances.NMR analysis highlighted presence of plasticizers in both COC body and elastomer stopper but concentrations were below the 1000µg/g threshold described by the REACH regulation.Cefuroxime solution (1mL at 10mg/mL) was stable 365 days at -20°C. Midazolam solution (50mL at 1mg/mL) and noradrenaline solution (20mL and 50mL at 0.2mg/mL ; 50mL at 0.5mg/mL) were stable 365 days at 5°C. Same results were achieved in polypropylene syringes.Crystal® vials were not subject to sorption phenomena. There was no risk of plasticizer migration. Metacresol permeation risk was controlled using the new stopper. Packaging of alkaline solution must be monitored. Solutions of cefuroxime (-20°C), midazolam and noradrenaline (5°C) are stable 1 year. In regards to storage in polypropylene syringes, Crystal® vials could be prefered for their automated aseptic filling process and for logistic considerations particularly taking into account the space gains.

Les nanoémulsions sont des formulations intéressantes pour des applications telles que les cosmétiques, les produits pharmaceutiques et les produits alimentaires. Elles peuvent être produites par des techniques à basse ou haute énergie. Dans ce travail, un procédé impliquant une pression modérée, l'émulsification membranaire par prémix a été proposé comme alternative. Des nanoémulsions huile-dans-eau (H/E) et eau-dans-huile (E/H) ont été produites avec une installation à l'échelle pilote composée d'un pousse-seringue à haute pression et d'une membrane Shirasu Porous Glass (SPG). Tout d'abord, l'influence des nombreux paramètres de procédé et de composition sur la taille des gouttelettes et la pression résultante a été étudiée avec des compositions modèles afin d'optimiser la production. Ainsi, des nanoémulsions H/E d'environ 260 nm et E/H d'environ 600 nm ont été produites avec succès. Puis, le montage a été utilisé pour produire des nanoémulsions de compositions spécifiques, des nanoémulsions H/E et E/H stabilisées avec des tensioactifs polypeptidiques et une nanoémulsion H/E adaptée à l'injection. Enfin, le procédé développé a été comparé à deux procédés à haute énergie traditionnels, le microfludiseur et les ultrasons en termes de taille des gouttelettes et de conservation d'actifs. Aucune différence entre les procédés n'a été observée en ce qui concerne la préservation de l'acif choisi. Cependant, en ce qui concerne la taille, les nanoémulsions produites par les membranes ont présenté des gouttelettes monodisperses de 335 nm par rapport aux autres procédés qui ont produit des nanoémulsions d'environ 150 nm de taille moyenne mais contenant aussi des gouttelettes de taille micrométrique détectées par diffraction laser et microscopie optique. Pour cette raison, les nanoémulsions produites par procédé membranaire conviennent également pour des applications parentérales sans étape de filtration supplémentaire / Nanoemulsions are interesting carriers for applications such as cosmetics, pharmaceutical and food. They are produced usually by low or high energy techniques. In this work, a process involving moderate pressure, premix membrane emulsification (PME) was proposed as an alternative. Oil-in-water (O/W) and water-in-oil (W/O) nanoemulsions were produced with a pilot scale set-up composed of a controlled high pressure syringe pump and Shirasu Porous Glass (SPG) membrane. First, the influence of process and composition parameters on droplet sizes and pressures was extensively studied with model compositions to optimize the production. Thus, nanoemulsions down to 260 nm for O/W and around 600 nm for W/O were successfully produced. Then, the set-up was used to produce nanoemulsions of specific compositions: O/W and W/O nanoemulsions stabilized with polypeptidic surfactants and O/W nanoemulsions suitable for injection. Finally, the set-up developed was compared to two traditional high energy processes, microfludizer and ultrasound in terms of droplet size and active preservation. No real difference between the three processes was seen on active preservation with the model active chosen. However, regarding droplet size, PME produced monodispersed droplets of 335 nm compared to the other processes which produced nanoemulsions of around 150 nm but with the presence of micron size droplets detected by laser diffraction and optical microscopy. Therefore, PME nanoemulsions are also suitable for parenterals applications with no additional filtration step required

Submitted by Cleide Dantas (cleidedantas@ufpa.br) on 2014-05-05T16:14:05Z No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Tese_ConvergenciasDivergenciasSistema.pdf: 1227100 bytes, checksum: d125e8886cbadf7407faa9dd760bd529 (MD5) / Approved for entry into archive by Ana Rosa Silva (arosa@ufpa.br) on 2014-05-08T14:22:35Z (GMT) No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Tese_ConvergenciasDivergenciasSistema.pdf: 1227100 bytes, checksum: d125e8886cbadf7407faa9dd760bd529 (MD5) / Made available in DSpace on 2014-05-08T14:22:35Z (GMT). No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Tese_ConvergenciasDivergenciasSistema.pdf: 1227100 bytes, checksum: d125e8886cbadf7407faa9dd760bd529 (MD5) Previous issue date: 2010 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / A proposta desta pesquisa foi identificar convergências e divergências nas crenças, metas de socialização e práticas de cuidados parentais em dois contextos ecológicos amazônicos, buscando analisar a relação entre fatores biológicos e ecoculturais. Participaram da pesquisa 99 mães de duas localidades: Belém – capital (CEU – contexto ecológico urbano – N=50) e Santa Bárbara (CENU – contexto ecológico não urbano – N=49), com idade superior a 18 anos com pelo menos uma criança com idade entre 0 e 6 anos . Os dados foram coletados por meio de entrevistas em que se aplicavam questionários e para análise destes foram utilizados critérios estatísticos e o respaldo teórico da Psicologia Evolucionista. Os dados indicaram que as comunidades estudadas em relação aos dados socio-demográficos diferem de forma sistemática uma da outra, em função dos níveis de desenvolvimento tecnológico e diferentes organizações para o contexto social. O mundo social das crianças de CENU e de CEU representam contextos desenvolvimentais que oferecem diferentes oportunidades. A literatura aponta que as trocas sociais estabelecidas entre crianças e cuidadores podem refletir sobre o desenvolvimento da criança em relação ao aspecto cognitivo, emocional e social. Os resultados encontrados indicam que as mães de CEU apesar de terem sido mais autônomas que as de CENU, foram ao mesmo tempo mais relacionais que as mesmas, demonstrando que talvez as mudanças que atingem os contextos, ocorram com mais rapidez em ambientes urbanizados. Além disso, os dados parecem indicar que os contextos por estarem em um momento de transição passam de um modelo interdependente para um modelo autônomo-relacional. Entretanto, foi possível perceber que as idéias que as mães possuem sobre a importância de determinadas ações nem sempre refletem suas práticas. Em relação aos cuidados primários as mães parecem valorizar e realizar de igual modo. Em relação ao Contato corporal as mães de CENU valorizam mais que as mães de CEU. Em estimulação corporal o resultado foi bastante interessante, pois os itens que CEU deu mais importância e praticou menos, foram os mesmos itens em que CENU deu menos importância e praticou mais. No que tange a estimulação por objeto as mães de CEU dão mais importância as praticas do que as realizam e vice-versa acontece em CENU. Ao sistema face-a-face foi atribuída maior importância pelas mães de CEU. Os dados apresentados sugerem que as mães de ambos os contextos estão utilizando estratégias parentais distais e proximais ao mesmo tempo, ou seja, desejam que seus filhos se tornem auto-suficientes, mas também desejam que os mesmos sejam respeitosos e obedientes. Além disso, os resultados aqui encontrados confirmam que os quatro sistemas descrevem as experiências interacionais das crianças e expressam a ênfase cultural de combinações e estilos particulares. Não foram encontradas diferenças marcantes nas crenças e práticas de cuidados maternos entre CEU e CENU, o que nos levou a considerar que as crenças parentais, em consequência à adaptação ao contexto, variaram menos conspicuamente nas cidades escolhidas do que entre outras cidades examinadas em outros estudos. Outra questão bastante interessante encontrada foi o resultado relativo ao nível de instrução das mães de CEU e a valorização de metas relacionais, pois de acordo com a hipótese na literatura o nível educacional das mães torna-se uma variável importante em relação às metas de socialização. CENU demonstrou uma tendência para metas e práticas relacionais. Isso se deve ao modo de vida mais próximo do protótipo de interdependência que este contexto apresenta, no qual as mães tendem a valorizar as normas e regras determinadas pela família ou grupo ao qual pertencem. Os resultados referentes ao CEU não confirmaram a hipótese de que ele apresentaria mais metas e práticas voltadas para a independência, sendo percebido que Belém apresenta tanto características do modelo de independência como de interdependência. Presume-se que CEU e CENU são contextos que passam por mudanças, uma vez que as mães podem acreditar em uma coisa e na realidade agir de outra forma. / The purpose of this research was to identify convergecies and divergencies in ethnotheories, socialization goals and parenting practices in two Amazonian ecological contexts, trying to analyze the relationship between biological and ecoculturals factors. Ninety nine mothers participated of the research: Fifty from Belém ( CEU – Urban context) and forty nine from Santa Barbara ( CENU – non urban context) . The mothers aged over 18 years with at least one child aged between 0 and 6 years. Data were collected through interviews and questionnaires were applied for the data analysis were applied statistical criteria and theoretical support of Evolutionary Psychology. The data indicated that the communities studied in relation to sociodemographic data differ systematically from one another, depending on the levels of technological development and organizations of social context. The social world of children from CEU and CENU represent developmental contexts that offer different opportunities. The literature indicates that the established social exchanges between children and caregivers can reflect on the child's development in relation to the cognitive, emotional and social. The results indicate that mothers of CEU despite being more autonomous than those of CENU were relational too, showing that perhaps the changes that affect the context, occurring faster in urbanized environments. Moreover, the data seem to indicate that the contexts are transition from the interdependent model to an autonomousrelational model. However, it is noted that the ideas that mothers have about the importance of certain actions do not always reflect their practices. In relation to primary care mothers seem to appreciate and perform equally. Mothers of CENU value more than mothers of CEU the body contact. The results directed to body stimulation were very interesting, because the items that were more important and charged by CEU mothers were the same that were less important and charged by CENU mothers. With respect to object stimulation CEU mothers give more importance to the practices than carrying out the practices and vice versa happens in CENU. The mothers of CEU attributed greater importance to face face system. Our data suggest that mothers of both contexts are using distal and proximal parenting strategies at the same time, or want their children to become self-sufficient, but also want them to be respectful and obedient. Moreover, our findings confirm that four systems describe the interactional experiences of children and express the cultural emphasis of particular styles and combinations. There were no significant differences in beliefs and practices between CEU and CENU mothers, which led us to consider that parental beliefs, therefore adapt to context, varied less conspicuously in the selected cities than in other cities examined in other studies. Another issue found that was very interesting was about the CEU mothers’ educational level and the valorization of relational goals, because according literature, the educational level of mothers becomes an important variable in relation to socialization goals. CENU has shown a tendency to relational practices and relational goals. This context provides a way of life closer to the interdependence model, in which mothers tend to value the norms and rules determined by the family or group to which they belong. The hypothesis that practices and goals of CEU would be more independent could not be confirmed, so we realized that this context features both independent and interdependent models. It was assumed that CENU and CEU contexts are undergoing changes, since mothers may believe one thing and actually doing otherwise.

Introduction : Les enfants prématurés ont la particularité de naître alors que leur développement est souvent incomplet et nécessite la mise en œuvre de soins intensifs visant à poursuivre leur croissance en dehors de l’environnement utérin. Souvent cependant, le stade développemental de l’enfant ne lui permet pas d’assimiler une alimentation entérale du fait de l’immaturité de son système digestif. Le recours à une voie centrale délivrant les nutriments assurant le développement devient alors une nécessité. Ce type de nutrition, appelée nutrition parentérale (NP, ou total parenteral nutrition TPN), permet l’administration de molécules simples, directement dans le sang du prématuré. Il n’est toutefois pas exempt de risques puisqu’exposée à la lumière, la NP peut s’oxyder et générer des molécules oxydantes telles que des hydroperoxydes lipidiques susceptibles de se fragmenter par la suite en hydroxy-alkénals. Ceci devient problématique au vu de l’immaturité des systèmes de défenses antioxydants du nouveau-né prématuré. L’utilisation prolongée de la NP est d’ailleurs à l’origine de maladie hépatiques dans lesquelles le stress oxydant et la nécro-inflammation sont des composantes majeures. Nous avons émis l’hypothèse que l’infusion chez les enfants prématurés, d’aldéhydes d’origine lipidique est en relation avec le développement du stress oxydant et de l’inflammation hépatique. Objectif : Notre étude a consisté à évaluer la relation entre les quantités d’hydroxy-alkénals dans la NP et les effets hépatiques engendrés sur les marqueurs de stress oxydant et les voies de signalisation responsables d’une induction de processus inflammatoire. Dans ce but, nous avons cherché à mesurer la peroxydation lipidique dans l’émulsion lipidique de la NP et la conséquence de l’infusion en continue d’hydroxy-alkénals sur les marqueurs de stress oxydant, sur la voie de signalisation médiée par le Nuclear Factor κB et sur le déclenchement du processus inflammatoire hépatique. A la suite de ce travail, nous avons également travaillé sur des alternatives à la photoprotection, qui est la seule méthode réellement optimale pour réduire la peroxydation des lipides de la NP, mais cliniquement difficilement praticable. Résultats : Nos résultats ont mis en évidence la génération de 4-hydroxynonenal in vitro dans la NP, ce phénomène est augmenté par une exposition lumineuse. Dans ce cadre, nous avons montré l’inefficacité de l’ajout de multivitamines dans l’émulsion lipidique comme alternative à la photoprotection. Dans la validation biologique qui a suivi sur un modèle animal, nos résultats ont permis de démontrer que l’augmentation des adduits glutathion-hydroxynonenal était imputable à l’augmentation de 4-hydroxynonenal (4-HNE) dans la NP, et non à une peroxydation endogène. Nos données indiquent que la probable augmentation hépatique des niveaux de 4-HNE a conduit à une activation du NFκB responsable de l’activation de la transcription des gènes pro-inflammatoires du Tumour Necrosis Factor-α (TNF-α) et de l’interleukine-1 (IL-1). Nous avons alors évalué la capacité d’une émulsion lipidique enrichie en acides gras polyinsaturés (AGPI) n-3 à baisser les concentrations de 4-HNE dans la NP, mais également à moduler le stress oxydant et les marqueurs pro-inflammatoires. Enfin, nous avons démontré, en collaboration avec l’équipe du Dr Friel, que certains peptides isolés du lait humain (par un processus mimant la digestion) permettent également une modulation du stress oxydant et du processus inflammatoire. Conclusion : Le stress oxydant exogène issu de la NP a conduit par activation de facteurs de transcription intra-hépatiques au déclenchement d’un processus inflammatoire potentiellement responsable du développement de maladies hépatiques reliées à la NP telle que la cholestase. Dans ce sens, les AGPI n-3 et les peptides antioxydants peuvent se poser en tant qu’alternatives crédibles à la photoprotection. / Introduction: Premature infants usually born before full term require intensive care to continue to grow up outside the uterine environment. Premature newborns are born with gastrointestinal systems that are too immature to absorb nutrients safely. Therefore they receive their initial nutrients through intravenous feeding, called total parenteral nutrition which delivers simple nutrients directly into bloodstream. However, light exposed-TPN can generate oxidant molecules such as lipid hydroperoxides, which can potently break up into hydroxy-alkenals. Prolonged use of TPN is also a cause of liver disease in which oxidative stress and necro-inflammation are major components. Thus, we hypothesize that lipid aldehydes contained in TPN are associated with oxidative stress and hepatic inflammation developments. Objectives: The aim of our study is to assess the relationship between quantities of hydroxyl-alkenals generated in TPN and effects on oxidative stress biomarkers and cell-signalling pathways molecules implicated in hepatic inflammation induction. To this end, we measure lipid peroxidation in the TPN lipid emulsion in and the consequence of continuous infusion of hydroxy-alkenals on markers of oxidative stress, on cell-signaling pathway mediated by the NFkB, and on liver inflammation induction. Following these data, we also worked on alternatives of photoprotection, which is the only optimal method for preventing lipid peroxidation, but unfortunately clinically impractical. Results: In vitro studies have highlighted the generation of 4-HNE in the TPN, increased under light exposure. In this context, we have demonstrated that the addition of multivitamins in the lipid emulsion cannot be a valuable alternative to photoprotection. Concerning the biological validation in our guinea pig animal model, our results demonstrated that the increase of GS-HNE adducts was due to increased 4-HNE in the TPN, and does not provide from endogenous peroxidation. Our data also indicate that the increase of hepatic 4-HNE led to an activation of NFkB, responsible for the activation of the transcription of proinflammatory genes TNF-α, IL-1. In the next study, we have evaluated the ability of a lipid emulsion enriched with n-3 polyunsaturated fatty acids (PUFA) to reduce 4-HNE concentrations generated in TPN, and to modulate oxidative stress markers and pro-inflammatory process on the same animal model. We also have demonstrated, in collaboration with Dr Friel’s team, that two antioxidant peptides (derived from a process mimicking digestion process of human milk) allow also a modulation of oxidative stress and inflammatory process in the liver. Conclusion: This form of exogenous oxidative stress from the TPN led to an inflammatory process resulting from the activation of intrahepatic transcription, which is potentially responsible of liver disease development such as cholestasis. In this sense, the n-3 PUFA and antioxidant peptides may arise as a valuable alternative of photoprotection.

Problématique : Le glutathion est une molécule clé de la défense antioxydante. Chez les enfants sous nutrition parentérale (NP), particulièrement les nouveau-nés, sa concentration tissulaire est anormalement basse. Puisque la capacité de synthèse de glutathion est adéquate, un déficit en cystéine, le substrat limitant, est soupçonnée. À cause de son instabilité en solution, la cystéine est peu présente en NP; la méthionine étant le précurseur endogène de cet acide aminé. L’activité de la méthionine adénosyltransférase (MAT), une enzyme essentielle à la transformation de la méthionine en cystéine, est facilement inhibée par l’oxydation. L’hypothèse : Le faible taux de glutathion chez les enfants sous NP est causé par l’inhibition de la MAT par les peroxydes contaminant ces solutions nutritives. Objectif: Mesurer l’impact d’une infusion de NP et de H2O2 sur l’activité hépatique de MAT en relation avec le niveau de glutathion. Méthode : Un cathéter est placé dans la jugulaire droite de cobayes de trois jours de vie. Quatre groupes sont comparés:1- Témoin (animaux aucune manipulation, sans cathéter) 2)-(animaux nourris normalement et le cathéter (noué)); 3) NP (animaux nourris exclusivement par voie intraveineuse (acides aminés + dextrose + lipides + vitamines + électrolytes), cette solution génère environ 400 µM de peroxyde. 4) H2O2 (animaux nourris normalement et recevant via le cathéter 400 µM de H2O2). Après quatre jours, le foie et le sang sont prélevés pour la détermination du glutathion, potentiel redox et l’activité de MAT, glutathion peroxydase et glutathion reductase. Résultats : L’activité de MAT est plus faible dans les groupes NP et H2O2. Le potentiel redox du foie et dans le sang est plus oxydé dans le groupe NP. Tandis que la concentration de GSSG du foie est plus élevée dans le groupe NP. Ainsi la concentration de GSH dans le sang et foie est plus faible dans les NP et H2O2 Discussion: La relation entre l’inhibition de MAT et le stress oxydant observée dans le groupe NP pourrait bien expliquer la perturbation du système glutathion observée chez les nouveau-nés prématurés. / Introduction: The low glutathione level in premature newborns can partly explain the high incidence of complications associated with oxidative stress in this population. Since the synthetic activity of glutathione is mature, a lack of substrate, especially cysteine, is suspected. Methionine provided by their parenteral nutrition (PN) is the in vivo precursor of cysteine. Since, methionine adenosyltransferase (MAT), the first enzyme in methionine transformation, has redox sensitive cysteines, we hypothesize that peroxides contaminating PN inhibit the activity of MAT, leading to a lower availability of cysteine for glutathione synthesis. Methods: At 3 days of life, guinea pigs a catheter fixed in jugular vein, the animals were separated in 4 groups: 1) Control: animals without any treatment and no catheter 2) sham: animals fed regular chow, a node closed their catheter; 3) PN: animals fed exclusively with parenteral nutrition (dextrose, amino acids, fat, vitamins) containing about 400 µM peroxides; 4) H2O2: animals fed regular chow and received continuously 400 µM H2O2 through the catheter. Four days later, liver and blood were sampling for determination of reduced glutathione (GSH), oxidized glutathione (GSSG), redox potential and activities of MAT, glutathione peroxidase, and glutathione reductase. Results: MAT activity was lower in groups receiving PN or H2O2. Liver redox was more oxidized in PN group whereas blood redox was more oxidized in PN and H2O2. Liver and blood GSH is lower in PN and H2O2. Liver GSSG was higher in PN. Conclusion: The inhibition of MAT in the PN group could explain the disruption of the glutathione system observed in premature infants. Furthermore the impact of a lower activity of MAT on GSH level is observed in liver and blood. Suggesting that the hepatic synthesis of GSH is insufficient to maintain its own level of glutathione and sustain the rate of export.

Introduction: Le supplément d’oxygène et la nutrition parentérale (NP) sont les deux sources majeures de stress oxydant chez le nouveau-né. Lors de la détoxification des oxydants, le potentiel redox du glutathion s’oxyde. Notre hypothèse est que le supplément d’oxygène et la durée de la NP sont associés à un potentiel redox plus oxydé et à une augmentation de la sévérité de la dysplasie bronchopulmonaire (DBP). Patients et Méthodes: Une étude observationnelle prospective incluant des enfants de moins de 29 semaines d’âge gestationnel. Les concentrations sanguines de GSH et GSSG à jour 6-7 et à 36 semaines d’âge corrigé étaient mesurées par électrophorèse capillaire et le potentiel redox était calculé selon l’équation de Nernst. La sévérité de la DBP correspondait à la définition du NICHD. Résultats: Une FiO2≥ 25% au 7ième jour de vie ainsi que plus de 14 jours de NP sont significativement associés à un potentiel redox plus oxydé et à une DBP plus sévère. Ces relations sont indépendantes de l’âge de gestation et de la gravité de la maladie initiale. La corrélation entre le potentiel redox et la sévérité de la DBP n’est pas significative. La durée de la NP était responsable de 15% de la variation du potentiel redox ainsi que de 42% de la variation de la sévérité de la DPB. Conclusion: Ces résultats suggèrent que l’oxygène et la NP induisent un stress oxydant et que les stratégies visant une utilisation plus judicieuse de l’oxygène et de la NP devraient diminuer la sévérité de la DBP. / Introduction: oxygen supplementation and total parenteral solution (TPN) are two main clinical practices that sustain oxidative stress. Glutathione is a key molecule that detoxifies peroxides resulting in a more oxidized redox potential. We hypothesize that O2 supplementation and longer TPN duration are associated with both more oxidized redox potential and more severe bronchopulmonary dysplasia (BPD). Patients and methods: A prospective observational study including infants of less than 29 weeks gestational age. GSH and GSSG from whole blood sampled on day 6-7 and at 36 weeks of corrected age (CA) were measured by capillary electrophoresis and redox potential was calculated using Nernst equation. BPD was classified according to NICHD guidelines. Results: There was a significant association between FiO2 ≥ 25% on day 7 of life and TPN duration longer than 14 days and both more oxidized redox potential and more severe BPD. TPN duration explained both 15 % of total variation observed in redox potential and 42 % of total variation in BPD severity. These associations remained significant after adjustment for gestational age and illness severity. The relation between the severity of BPD and the redox potential in blood was not significant. The statistic power (1-β) to show an effect of redox potential on severity of BPD was 52%. Conclusion: Both redox potential of glutathione and BPD severity are both associated with early O2 supplement and TPN. Strategies targeting judicious use of O2 supplement and either decreasing the duration or using safer formulation of TPN are expected to help reducing BPD.

This diploma thesis deals with the issue of life quality of patients taking home parenteral nutrition. Its intention is to present the achieved life quality scores in the individual domains of physical and mental health in comparison with general population and to point out the differences in what dimensions the life quality of these patients is particularly affected. The theoretical section describes the present situation of the issue of home parenteral nutrition in the CR and the system of the care about the patients. The chapter about indications and contraindications to HPN is elaborated in more detail. The possibilities of the long-term vascular accesses, the care about them are also mentioned here, and particularly the role of a nurse in patient education in transferring parenteral nutrition to the domestic environment. The problems with long-term parenteral nutrition resulting from mechanic, metabolic and septic complications are also outlined. Parenteral nutrition failure is the most serious problem, which is why a chapter on small intestine transplantation as the last resort to save a patient with combined failure of intestine and nutrition is included. The second part of the theoretical section describes the life quality. I wanted to define the nature of this unambiguously graspable a term, determinants affecting life quality are also mentioned here. The possibilities of life quality measurement and assessment and particularly the follow-up use of the obtained data are mostly summarized here. Goals and hypotheses: Two goals were set to meet the main purpose of the thesis: Goal 1: To find whether the life quality of patients on HPN differs from that of the general public. Goal 2: To find the spheres in which the life quality of patients on HPN is mostly affected. A zero hypothesis was set to achieve the goals: Life quality of patients on HPN does not differ from that of the general public. There is no statistically significant difference between men and women. Eight alternative hypotheses to each life quality domain followed: H1: Patients on HPN show lower life quality score compared to the general public in the sphere of physical functions. H2: Patients on HPN show lower life quality score compared to the general public in the sphere of physical roles' limitation. H3: Patients on HPN show lower life quality score compared to the general public in the sphere of emotional roles' limitation. H4: Patients on HPN show lower life quality score compared to the general public in the sphere of emotional limitation of social functions. H5: Patients on HPN show lower life quality score compared to the general public in the sphere of pain. H6: Patients on HPN show lower life quality score compared to the general public in the sphere of general mental health. H7: Patients on HPN show lower life quality score compared to the general public in the sphere of vitality. H8: Patients on HPN show lower life quality score compared to the general public in the sphere of general health perception. A quantitative method of collected data analysis was applied to the research implementation. The research was based on the standardized questionnaire SF-36 supplemented with questions dealing with identification of respondents, time consumption of their treatment and their consequent satisfaction at the beginning. The questionnaire was distributed to patients using home parenteral nutrition in specialized nutrition centres of the Thomayer Faculty Hospital in Prague, Faculty hospitals in Brno and Hradec Králové and also by electronic means through the website of the citizen association Life without Intestine. The obtained data were statistically evaluated and processed into illustrative tables and graphs.Detailed mapping of the problems of life quality and highlighting of the neglected spheres of life quality of patients using HPN are the outputs of the thesis. The results will be presented to the members of the workgroup for HPN within their.

Entre les actes chirurgicaux et l'état métabolique il existe des nombreuses interactions. D'un côté, la réponse catabolique majeure induite par la chirurgie viscérale peut être contrôlée par une supplémentation nutritionnelle précoce, ce qui diminuerait la morbidité et la mortalité postopératoire et aussi les durées d'hospitalisation. L'albuminémie préopératoire est un bon facteur prédictif de l'état nutritionnel postopératoire, corrélée avec un pronostic postopératoire inférieure. La préalbumine reflète de façon plus sensible l'évolution de l'état nutritionnel. Principale hormone orexigène, la ghréline présente une cinétique perturbée lors des périodes postopératoires avec une augmentation de sa sécrétion au moment de la reprise de la nutrition entérale et des concentrations postopératoires moyennes inférieures à celles normales. Ces observations posent la question de l'utilité d'un traitement par analogues de la ghréline. De l'autre côté, la chirurgie bariatrique peut corriger les perturbations métaboliques corrélées à l'obésité, mais son efficacité n'est pas absolue. Par contre, en utilisant certains critères clinique (âge, IMC, présence d'un diabète sucré) et biologiques (insulino-résistance, taux d'IGF1), cette efficacité devient prédictible pour les interventions d'insertion d'anneau gastrique / Surgical interventions can have several effects on the metabolic status. On one hand, the important catabolic response caused by major digestive surgery can be controlled through an early nutritional support, which could reduce the mortality, morbidity and also the duration of hospitalization. The preoperative albumin level is a reliable predictive factor of the postoperative nutritional status and correlates to a worse postoperative prognosis. The prealbumin reflects in the most sensitive manner the evolution of the nutritional status. As the main orexigen hormone, ghrelin has a disturbed cynetics in the postoperative period with an augmentation of its secretion corresponding to the reintroduction of the enteral nutrition and mean postoperative concentrations that are lower than normal. These observations raise the question of the utility o a ghreline analogues’ treatment. On the other hand, bariatric surgery could correct the metabolic disturbances associated to obesity, but its efficacy is not absolute. However, by using certain clinical (age, BMI, presence of a diabetes mellitus) and biological (insulin-resistance, IGF1 level criteria, this efficac can be redictable in the case of gastric banding