Introduction: Le principal objectif de cette étude est de mesurer l’effet du GDF-5 sur l’homéostasie du cartilage. Le GDF-5 est un gène de susceptibilité de l’OA faisant partie de la famille des BMPs et qui favorise la synthèse du cartilage. Le but de notre étude a été de déterminer l’effet du GDF-5 sur le métabolisme catabolique ainsi que sur l’équilibre global des chondrocytes, principalement au niveau de l’Aggrécan. Méthode : Des chondrocytes arthrosiques canins et humains OA ont été exposés au GDF-5. L’expression des ARNm et des protéines a été analysée afin d’évaluer la production de l’Aggrécan et le ratio Col-II/Col-I au niveau des facteurs anaboliques et du phénotype. Pour le catabolisme, l’expression et l’activité des aggrécanases ADAMTS-4 et ADAMTS-5 ont été mesurées. Les épitopes NITEGE et CTX-II ont aussi été quantifiés dans le liquide synovial canin après des injections intraarticulaires de GDF-5. Résultats : Le GDF-5 provoque une augmentation de l’activité cellulaire des chondrocytes canins et humains. Pour les ARNm et l’expression protéique, le GDF-5 augmente l’expression de l’Aggrécan alors que les facteurs cataboliques le diminuent. Le phénotype reste inchangé en présence du produit, sauf à haute dose où on augmente le ColI. L’activité des aggrécanases diminue puisque l’épitope NITEGE diminue alors que le CTX-II augmente dans l’articulation. Conclusion : En somme, les facteurs anaboliques du cartilage sont favorisés, alors que les facteurs cataboliques sont diminués par le GDF-5. Cette action double permet d’illustrer l’effet du GDF-5, le classant comme un potentiel médicament modifiant la maladie de l’OA qui mérite d’être étudiée. / Purpose: The objective of this study is to assess the effect of GDF-5 on cartilage homeostasis. GDF-5 is a susceptibility gene for OA and member of the BMP super family. Studies have shown that it can increase expression of anabolic factors in chondrocytes. Therefore, our study indentifies how GDF-5 influences this metabolism and the global homeostasis of chondrocytes, aiming mainly towards Aggrecan. Methods : Osteoarthritic (OA) chondrocytes from canine and human models were exposed to GDF-5. Protein expressions, along with mRNA expression were assessed in order to investigate Aggrecan production and the ratio of Col-II/Col-I, for the anabolic phenotype markers. The aggrecanases ADAMTS-4 and ADAMTS-5 and their global activity were assed for the catabolic factors. The NITEGE and CTX-II epitope were also measured in synovial fluid of Pond-Nuki dogs that received intraarticular GDF-5 injections. Results : GDF-5 increases chondrocyte cellular activity, in our canine and human models. Both mRNA and protein expression of the chondrocytes Aggrecan were increased and the aggrecanases expression and activity were decreased. Collagen ratio did not show a phenotype, except et high dosage where the Col-I production is induced. Aggrecanase activity was lowered while CTX-II was increased. Conclusion : In conclusion, the anabolic cellular activity of OA chondrocytes increases while the catabolic factors decrease in presence of GDF-5. This double action illustrates the global effect of GDF-5, identifying it as a potential disease modifying factor of OA that should be further investigated.

Artikulärer Knorpel weist ein geringes Selbstheilungspotential auf und aktuelle Methoden zur Therapie von Knorpeldefekten resultieren meist im Aufbau fibrösen, mechanisch inferioren Regenerationsgewebes. Deshalb wird stetig versucht neue Therapiemethoden zu entwickeln, die häufig auf Tissue Engineering basieren. Ein Ansatz sind biogedruckte Tissue Engineering Konstrukte, die es ermöglichen durch das „Eindrucken“ vitaler Zellen die komplexen zellulären Strukturen des Knorpelgewebes nachzubilden. Trotz vielsprechender Ergebnisse in vitro, scheitert die langfristige Regeneration von Knorpeldefekten mittels Tissue Engineering jedoch häufig an der mangelnden Fixierung und Integration der Konstrukte in das umliegende Gewebe. Besonders um biogedruckte Hydrogelkonstrukte, für die viele gängigen Fixierungsoptionen ungeeignet sind, für das Knorpel Tissue Engineering nutzbar zu machen, müssen daher neue Fixierungs- und Integrationsmethoden entwickelt werden. Als Ansatz zur Verbesserung der osteochondralen Integration von Knorpelersatzmaterialien wurde im Rahmen dieser Arbeit ein GDF-5 beladener Gewebekleber getestet. GDF-5, ein Knorpelmorphogenetisches Protein, kann die chondrogene Differenzierung sowie die Hypertrophie von Vorläuferzellen fördern. Durch den Einsatz eines GDF-5 beladenen Gewebeklebers sollen in den Defekt einwandernde, wirtseigene Progenitorzellen zur chondrogenen Differenzierung und Mineralisierung angeregt werden, mit dem Ziel eine mineralisierte Knorpelgewebsschicht zwischen der subchondralen Knochen-platte und dem implantierten Tissue Engineering Konstrukt zu generieren und so dessen Integration zu verbessern.

Det har forskats mycket på olika signalvägar i det mänskliga genomet, trotts detta finns det många frågetecken som kvarstår. Denna uppsats undersöker några av dem. Syfte: Undersöka förändringar i genuttryck och mRNA-nivåer för hypertrofi- (MRF4) translations- (5.8S & 18S) och proteolysreglerande gener (MuRF1 & GDF-8) efter en 10 veckor lång styrketräningsperiod hos kvinnor och män. Frågeställningar: (1) Finns det en förändring i total mängd RNA före och efter en 10 veckors styrketräningsintervention. (2) Finns det en förändring i uttryck av MRF4, 5.8S, 18S, MuRF1 samt GDF-8 efter en 10 veckors styrketräningsintervention. (3) Finns det en könsskillnad i förändringen av total mängd RNA samt aktivering av MRF4, 5.8S, 18S, MuRF1 och GDF-8 efter en 10 veckors styrketräningsintervention. Metod: Urvalet för analysen bestod av 16 otränade försökspersoner varav 8 var män och 8 var kvinnor. Försökspersonerna utförde unilateral styrketräning av nedre extremiteten under 10 veckor, under 2 av dessa veckor utfördes ocklusionsträning.  Träningsperiodiseringen var vågformig (70-90% av 1RM, 5-12 rep, 3 ggr/vecka). Muskelbiopsier togs i det arbetande benet före träningsperiodens start samt 3-7 dagar efter träningsperiodens avslut. Genuttryck analyserades med qPCR. Resultat: Det fanns ingen signifikant skillnad i förändring mellan män och kvinnors totala RNA eller genuttryck. Total RNA ökade signifikant (p<0,01) med 19,2 %. Kvinnorna hade en signifikant ökning (P<0,05) av RNA på 27,6 % medan männen hade en signifikant ökning (p<0,05) på 14 %. MRF4 hade en signifikant (P>0,05) procentuell ökning i genuttryck med 55,7 % och kvinnor för sig hade en signifikant (P>0,05) ökning på 64 %. GDF-8 ökade signifikant (P>0,05) med 55,5 % medan GAPDH ökade signifikant (P>0,05) för båda könen tillsammans med 70,6 % och för män med 87,8 %. MuRF1 och 5.8S hade inga signifikanta förändringar i genuttryck. Slutsats: Det verkar som att både män och kvinnor får en liknande procentuell förändring av total RNA och mRNA genuttryck 3-7 dagar efter en 10 veckors hypertrofistyrd styrketräningsperiod. För att mäta genuttryck av translationsgenen MRF4 verkar 3-7 dagar efter en 10 veckors styrketräningsperiod vara en tidpunkt då det fortfarande pågår hypertrofi av skelettmuskulaturen.  Av de proteolysreglerande generna GDF-8 och MuRF1 sågs en uppreglering av GDF-8 vilket skulle kunna vara ett tecken på att hypertrofin börjar hämmas. Ett oväntat fynd var att GAPDH visade sig vara olämplig som kontrollgen vid en styrketräningsintervention på 10 veckor och att 18S var väldigt stabil. Detta kan betyda att GAPDH inte skall användas vid längre styrketräningsinterventioner. / There have been much research on signaling pathways in the human genome, but there still remain many questions. This paper examines some of them. Aim: Investigate changes in gene expression and mRNA levels of hypertrophy (MRF4), translation (5.8S & 18S) and proteolysis regulating genes (GDF-8) after a 10-week strength training period in men and women. Research questions: (1) Is there a change in the total amount of RNA before and after a 10-week strength training intervention. (2) Is there a change in the expression of MRF4, 5.8S, 18S, Murf1 and GDF-8 after 10 weeks of strength training. (3) Is there a gender difference in the change of total RNA and the expression of MRF4, 5.8S, Murf1 and GDF-8 after a 10-week long strength training intervention. Method: The sample for analysis consisted of 16 untrained subjects, of whom 8 were men and 8 were women. The subjects performed unilateral resistance training of lower extremities for 10 weeks, during two of these weeks blood flow restriction training were performed. The training was undulating (70-90% of 1RM, 5-12 cord, 3 times / week). Muscle biopsies were taken from the working leg before the start and 3-7 days after the training period. Gene expression was analyzed by qPCR. Results: There was no significant gender difference in total RNA or gene expression. Total RNA was significantly increased (p <0.01) with 19.2 %. The women had a significant increase (P <0.05) of RNA at 27.6 %, while the men had a significant increase (p <0.05) at 14 %. MRF4 had a significant (P> 0.05) percentage increase in gene expression by 55.7 %, and women had a significant (P> 0.05) increase of 64 %. GDF-8 increased significantly (P> 0.05) with 55.5 %, while GAPDH increased significantly (P> 0.05) for both sexes with 70.6 % and for men with 87.8 %. Murf1 and 5.8S had no significant changes in gene expression. Conclusions: It seems that both men and women experience a similar percentage difference of total RNA and mRNA gene expression 3-7 days after a 10 weeks long strength training period. To measure the gene expression of MRF4 3-7 days after a 10-week weight-training period seems to be a time when there still is a anabolic responses in the skeletal muscle. Of the proteolysis regulating genes GDF-8 and Murf1 there was an upregulation of GDF-8, which could be a sign that the inhibition of hypertrophy started. An unexpected finding is that GAPDH was found to be unsuitable as a control gene at a strength training intervention at 10 weeks and rRNA 18S was very stable, which could mean that GAPDH should not be used as control gene in longer strength training studies.

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Este trabalho descreve, analisa e compara, no espanhol peninsular (EP) e no português brasileiro (PB), as estratégias de ordenação dos constituintes de Propriedade Configuracional (argumentais) ou correferenciais a eles, e que portam funções pragmáticas (Tópico, Foco e Contraste) ou retóricas (Orientação e Esclarecimento). O referencial teórico desta pesquisa é o da Gramática Discursivo-Funcional - GDF (HENGEVELD & MACKENZIE, 2008; MACKENZIE, 2012; PEZATTI, 2014, 2017) e o dos estudos comparativos entre o PB e o E (GONZÁLEZ, 1994, 2008, 2014; GROPPI, 2004; FANJUL, 2014; PINHEIRO-CORREA, 2015, 2018). A unidade de análise selecionada é o Ato Discursivo, o que significa que são considerados tanto constituintes oracionais como extraoracionais. Dentre os oracionais, os analisados são os de sentenças simples de Ilocuções Declarativas e Interrogativas e de Estado-de-Coisas [+dinâmicos]. Com base nos objetivos e no referencial teórico desta pesquisa, se analisa dois subcorpora de língua oral coloquial, um extraído do projeto C-ORALROM, que contempla a variedade de Madri, e outro do C-ORAL-BRASIL, que contempla a variedade mineira. A hipótese central que se tratou de comprovar é a de que, para enunciados que não sejam contrastivos, o EP se caracteriza pela não topicalização dos Sujeitos e topicalização dos Objetos, em um processo oposto ao que ocorre em PB, que prefere topicalizar os Sujeitos e não topicalizar os Objetos. Considerando a perspectiva discursivofuncional elegida neste estudo, toma-se por base que cada posição do Enunciado tende a ser ocupada por diferentes tipos de constituintes. Para analisar quais são as preferências, motivações e limitações para os posicionamentos de constituintes com as funções pragmáticas e retóricas supracitadas, são investigados fatores como (i) a relação entre determinação/especificidade/genericidade e Tópico/Foco, e as consequências dessa relação no Nível Morfossintático de cada língua, tais como duplicações por clíticos ou por pronomes fortes, não realização versus realização de Sujeitos pronominais, etc.; (ii) estruturas especiais para a marcação de Foco e de Contraste em ambas as línguas, principalmente as obtidas exclusivamente pela ordenação; (iii) os tipos de respostas dadas às sentenças Interrogativas Polares e a posição dos constituintes nas Interrogativas de Conteúdo. A tese está dividida em duas grandes partes: a primeira concentra as bases do referencial teórico escolhido e apresenta os procedimentos metodológicos adotados; a segunda mescla teoria, descrição e análises dos dados dos corpora. Os resultados evidenciam variadas diferenças entre as línguas, tanto no que se refere à ordenação quanto a outras estratégias usadas, a depender da função sintática dos constituintes aos quais se atribuem as funções pragmáticas e retóricas. Algumas vezes os mesmos recursos estão disponíveis em ambas as línguas, mas diferem entre si pelo grau de preferencialidade entre uns e outros. Outras vezes, uma língua recorre a estratégias que não estão disponíveis na outra. De maneira geral, o sistema de esquemas de posições proposto pela GDF possibilitou entender que estruturas superficialmente iguais podem ser diferentes em sua funcionalidade. / This work describes, analyzes and compares, in Peninsular Spanish (PS) and in Brazilian Portuguese (PB), the ordering strategies of the Configurational Property constituents (argumental) or co-referential to them and that carry pragmatic functions (Topic, Focus and Contrast) and rhetorical functions (Orientation and Clarification). The theoretical reference of this research is the Functional Discourse Grammar - FDG (HENGEVELD & MACKENZIE, 2008; MACKENZIE, 2012; PEZATTI, 2014, 2017) and the comparative studies between BP and S (GONZÁLEZ, 1994, 2008, 2014; GROPPI, 2004; FANJUL, 2014; PINHEIRO-CORREA, 2015, 2018). The unit of analysis selected is the Discourse Act, which means that they are considered both in the clause scope constituents and in the linguistic expression constituents. Among the clause scope ones, the analyzed are those of simple sentences of Declarative and Interrogative Ilocutions and of State-of-Affairs [+dynamic]. Based on the objectives and the theoretical reference of this research, what are analyzed are two subcorpora of colloquial oral language extracted from the project C-ORAL-ROM, that contemplates the variety of Madrid, and CORAL- BRASIL, that contemplates the Minas Gerais States variety. The central hypothesis that was sought to be confirmed was the one that, for statements that are not contrastive, the PS is characterized by the non-topicalization of the Subjects and by the topicalization of the Objects, in an opposite process to what occurs in BP, which tends to topicalize the Subjects and nontopicalize the Objects. Considering the functional discourse perspective chosen in this study, it is assumed that each position of the Statement tends to be occupied by different types of constituents. In order to analyze the preferences, motivations and limitations for constituents positions with the aforementioned pragmatic and rhetorical functions, factors such as (i) the relation between determination/specificity/genericity and Topic/Focus are investigated, as well as the consequences of this relation in Morphosyntactic level of each language, such as clitic doublings or strong pronouns, no materialization or materialization of pronominal Subjects, etc.; (ii) special structures for Focus and Contrast marking in both languages, especially those obtained exclusively by ordination; (iii) the types of responses given to the Polar Interrogative sentences and the position of the constituents in the Content Interrogatives. The thesis is divided in two major parts: the first one concentrates the bases of the chosen theoretical reference and methodological questions and the second one merges theory, description and results of the analysis of data obtained from corpora. The results show several differences between languages, both in terms of ordering and other strategies used, depending on the syntactic function of the constituents to which pragmatic and rhetorical functions are attributed. Sometimes the same resources are available in both languages, but they differ by the degree of preference between them. At other times, one language resorts to strategies that are not available in the other. In general, the system of position schemes proposed by FGD has made it possible to understand that apparently equal structures on the surface may be different in their functionality.

La circulation extracorporelle compte parmi les progrès techniques majeurs associés à la chirurgie cardiaque. Elle constitue aussi l’une des causes de complications principales car responsable d’une réponse inflammatoire généralisée qui résulte de la conjugaison des effets du stress oxydant et des cytokines libérés, contribuerait à la dysfonction multi-organe aboutissant aux complications myocardiques et rénales survenant au cours des périodes per- et postopératoires. La première partie de notre travail avait pour objectif d’explorer l’évolution des taux circulants du GDF-15, cytokine associée au stress oxydant et à l’inflammation, dans ce contexte de chirurgie cardiaque. Notre étude prospective a démontré pour la première fois que cette procédure est accompagnée de l’augmentation du GDF-15 dont les taux plasmatiques sont associés aux lésions postopératoires cardiaques et rénales.L’évaluation du risque opératoire repose sur un ensemble de scores dont le calcul est basé essentiellement sur des caractéristiques cliniques. Ces scores présentent toutefois un certaines limitations. Chez les patients « médicaux » atteints de pathologies cardiovasculaires la stratification du risque est définie en associant des caractéristiques cliniques à l’évaluation des taux circulants de biomarqueurs. L’objectif de cette seconde partie a donc été de mettre en évidence le pouvoir prédictif du GDF-15 en tant que biomarqueur circulant dans la survenue de complications rénales au cours de la chirurgie cardiaque sous CEC. Nous avons mis en évidence que les patients présentant des taux préopératoires élevés de GDF-15 sont à risque de développer une insuffisance rénale aigue postopératoire. / Ischemic cardiac diseases are the most frequent and deleterious pathologies leading to important cardiovascular-related mortality worldwide. One of the alternative therapies consists to treat these patients using cardiac surgery. Cardiopulmonary bypass was developed to greatly improve this surgical procedure. However, some adverse effects can occur during cardiac surgery associated with cardiopulmonary bypass due to the inflammatory response. This phenomenon is the result of various mechanisms including oxidative stress and inflammatory cytokines which lead to multi-organ failure and then to myocardial and renal injuries occurring during the peri- and post-operative periods.The first part of this work was designed to evaluate in the context of cardiac surgery the kinetics of plasma GDF-15 levels, an oxidative stress and inflammation related cytokine. Our prospective study demonstrated for the first time the kinetic increase in plasma GDF-15 levels which were associated to postoperative cardiac and renal injuries.Currently, operative risk evaluation is based on score calculation including clinical criteria. These risk scores present some limitations. Concerning other cardiac patients out of surgical fields, the risk assessment is defined using clinical parameters and biomarkers evaluation (cardiac troponin, BNP, Nt-proBNP). Thus, we aimed to determine whether pre-operative GDF-15 as plasma biomarker could help to identify patients at high risk of renal injuries. We found that patients with the highest pre-operative plasma GDF-15 levels are at risk for post-operative acute kidney injury.

Philosophiae Doctor - PhD / The aim of this thesis is first, to evaluate various culture conditions to improve embryo development, and secondly, to analyze spent culture media for any biomarkers that may be predictive of embryo health. Single-step and sequential culture media were compared in both Planer and EmbryoScope™ incubators. Single-step media resulted in better blastocyst development compared to sequential media and the EmbryoScope™ incubation system showed slight improvements in embryo development than the Planer system. The benefits of supplementing the culture medium with either insulin or insulin-like growth factor 1 (IGF-1) or culturing in a 2% O2 environment, using two different strains of mice (hybrid and C57), as well as the suitability of these strains for quality control were compared. In insulin, hybrid embryos were slower to blastulate and had a lower blastocyst rate, whereas C57 embryos were slower to the morula and faster to blastocyst stages, and lower blastocyst rate than the controls. IGF-1 showed no difference in time-lapse morphokinetics (TLM) or blastocyst rates compared to controls in both hybrid and C57 embryos. Under 2% O2, hybrid embryos showed no significant difference in TLM up to the 8-cell stage, but slowed down afterwards, resulting in blastocysts with significantly lower cell counts than the 6% O2 group. The C57 embryos were slower to reach morula and expanded blastocyst, and had lower blastocyst rates in 2%O2 vs 6%O2. The C57 strain had significant slower overall embryo development for all time points than hybrid embryos in insulin, IGF-1 and ultra-low O2, as well as lower blastocyst rates. Measurement of growth differentiation factor 9 (GDF-9) and oxidation-reduction potential (ORP) in spent media as markers for embryo health were evaluated. Day 5 human blastocysts yielded higher pregnancy rates and GDF-9 levels in spent media compared to Day 6 blastocysts, but TLM parameters showed no impact on pregnancy outcome. In Day 6 blastocysts, the non-pregnant group showed significantly faster embryo development compared to the clinically pregnant group up to the 8-cell stage and start of blastulation. GDF-9 did not show any significant differences between non-pregnant and pregnant groups of Day 5 or Day 6 embryo transfers. ORP in spent media from good quality Day 3 embryos that developed into blastocysts were significantly higher than from those that did not, with no difference in control medium ORP. Spent media from arrested embryos showed lower ORP than their corresponding controls. Arrested embryos had slower development at syngamy, morula, blastulation and blastocyst stages. The single step medium in the EmbryoScope™ is the preferred choice for embryo culture. Insulin or IGF-1 media supplementation or 2% O2 culture did not provide any benefit to embryo development. The C57 mouse strain is more sensitive and may be better to detect changes in culture conditions, and therefore better model for quality control assays. GDF-9 values decrease from Day 5 to Day 6 which gives new insight to understanding the role of GDF-9 during embryogenesis. ORP in spent media indicate that embryos that developed into blastocysts did not contribute to ROS, but maintained ORP balance.

Le risque professionnel en France est géré par plusieurs régimes, le régime général et les régimes dits spéciaux (plus particulièrement le régime des militaires, des fonctionnaires de l'Etat, des agents EDF GDF, des non titulaires de l'Etat et des ouvriers d'Etat). Si cette singularité fut justifiée historiquement, elle ne l'est plus aujourd'hui. Le travail a en effet évolué depuis. Les conditions de travail d'un électricien du 19ème siècle, profession exercée par des salariés du secteur privé et public ne sont pas celles de celui du 21ème siècle. Le maintien en l'état de ces différents régimes n'est pas équitable. Selon son statut professionnel, le travailleur sera indemnisé de manière différente. Cette situation ne respecte pas un des principes fondamentaux de notre constitution : le principe d'égalité. Pour mettre fin au dysfonctionnement, l'uniformité paraît être la solution idéale. Or la mise en place d'un nouveau régime de risque professionnel risque de se heurter à l'attachement de leurs bénéficiaires à leur régime actuel. Qui dit nouveau régime, ne dit pas forcément destruction de l'existant. Il est proposé de créer une nouvelle branche aux côtés des autres branches de la Sécurité sociale. La branche du risque professionnel succèdera à la branche AT/MP. Mais à la différence de cette dernière, elle sera indépendante administrativement et autonome financièrement et elle permettra de meilleures reconnaissance et indemnisation des accidents ou des maladies liées au travail.