Submitted by Angela Maria de Oliveira (amolivei@uepg.br) on 2018-04-20T12:04:40Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Karla Miky.pdf: 833137 bytes, checksum: 4aac692850a86aaeabe93d6bbcf5af02 (MD5) / Made available in DSpace on 2018-04-20T12:04:40Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Karla Miky.pdf: 833137 bytes, checksum: 4aac692850a86aaeabe93d6bbcf5af02 (MD5) Previous issue date: 2018-03-09 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O óleo de linhaça tem se destacado como alimento funcional para humanos, sendo a fonte mais rica de ácido graxo α-linolênico. Quatro dietas isentas de farinha de peixe foram formuladas para serem isoproteicas (321,2 g/kg de proteína brua), isocalóricas (17,1 Mcal/kg de energia bruta) e isolipídicas (73,1 g/kg de lipídios totais), contendo duas fontes de óleos vegetais (óleo de soja ou óleo de linhaça) suplementadas em dois níveis (15 ou 30 g / kg). Foram distribuídos ao acaso 144 peixes (1076,3 ± 37,2 g) em esquema fatorial 2x2, em 12 gaiolas flutuantes de 1000 L cada, alimentadas manualmente até saciedade aparente. Observou-se maior aumento de peso, consumo de ração e melhor conversão alimentar em peixes alimentados com 30 g/kg de óleo de linhaça em comparação com peixes alimentados com 15 e 30 g/kg de óleo de soja. O rendimento de filé foi maior em peixes alimentados com 30 g/kg de óleo de linhaça em relação aos peixes alimentados com 30 g/kg de óleo de soja. Não foram observadas diferenças no peso corporal inicial, índice hepatosomático e composição próxima dos filés. Peixes alimentados com 30 g/kg de óleo de linhaça apresentaram maior teor de 18: 3 n-3 e menor teor de 18:2n-6 nos filés em comparação com peixes alimentados com 15 e 30 g/kg de óleo de soja. As maiores somas de SFA, MUFA e AG n-3 foram observados em filés de peixes alimentados com 15 g/kg de óleo de soja. Peixes alimentados com 30 g/kg de óleo de linhaça apresentaram maiores somas de ácidos graxos PUFA e de ácidos graxos n-3 nos filés. Filés de peixes alimentados com óleo de linhaça apresentaram menor relação de ácidos graxos n-6/n-3 em comparação com peixes alimentados com óleo de soja. A cor, capacidade de retenção de água, pH e a dureza dos filés não foram afetados. A adesividade dos filés analisada um e sete dias pós mortem foi maior em peixes alimentados com óleo de linhaça, enquanto observou-se menor mastigabilidade em filés de peixes alimentados com 30 g/kg de óleo de soja e de linhaça em relação aos peixes alimentados com 15 g/kg de óleo de soja e linhaça. Em conclusão, o óleo de linhaça demonstrou ser um alimento funcional como fonte de AG α-linolênico e para aumentar a relação de ácidos graxos n-6/n-3 nos filés. Além disso, recomenda-se 30 g/kg de óleo de 8 linhaça em dietas de terminação para melhorar o desempenho produtivo de tilápias do Nilo em terminação. / Linseed oil has emerging as functional food for human being one richest source of α-linolenic fatty acid. Four fishmeal-free diets were formulated to be isonitrogenous (321.2 g/kg of crude protein), isocaloric (17.1 Mcal/kg of gross energy) and isolipidic (73.1 g/kg of total lipids), containing two sources of vegetable oils (soybean oil or linseed oil) supplemented at two levels (15 or 30 g/kg). A hundred and forty-four fish (1076.3 ± 37.2 g) were distributed in a completely randomized in a 2x2 factorial scheme, into twelve 1000 L each floating cage, hand fed until apparent satiety and reared at 18 to 24 oC, during 6 wk. Higher weight gain, feed intake and improved feed conversion ratio were observed in fish fed 30 g/kg of linseed oil compared to fish fed 15 and 30 g/kg of soybean oil. Fillet yield of fish fed 30 g/kg of linseed oil was higher compared to observed in fish fed 30 g/kg of soybean oil. No differences on initial body weight, hepatosomatic index and proximate composition of fillets were observed. Fish fed 30 g/kg of linseed oil showed higher 18:3 n-3 and lower 18:2 n-6 content in the fillets compared to fish fed 15 and 30 g/kg of soybean oil. Higher sum of SFA, MUFA and n-3 FA were observed in fillets of fish fed 15 g/kg of soybean oil. Fish fed 30 g/kg of linseed oil showed higher sum of PUFA and sum of n-3 fatty acids in the fillets. Fillet of fish fed linseed oil showed lower ratio of n-6/n-3 fatty acids compared to fish fed soybean oil. Color, water holding capacity, pH and hardness of fillets were not affected. The adhesiveness of fillets analyzed at one and seven days post-mortem was increased in fish fed linseed oil, while lower chewiness was observed in fillet of fish fed 30 g/kg of soybean or linseed oil compared to observed in fish fed 15 g/kg of soybean or linseel oil. In conclusion, linseed oil was demonstrated to be functional food as α-linolenic source to enhance n-6/n-3 ratio of the fillets. In addition, linseed oil at 30 g/kg is recommended in finishing diets

Les traitements actuels de la leucémie myéloïde chronique (LMC) ne permettent pas d’éliminer la totalité des cellules leucémiques. Dans le but de développer un traitement curatif, il est donc nécessaire de parvenir à une meilleure compréhension des mécanismes sous-jacents des réponses partielles aux traitements, Dans ce travail, nous avons postulé qu’il existe des mécanismes de contrôle extrinsèques de la LMC pouvant influencer l’efficacité des différents traitements. Nous avons choisi d’étudier le rôle potentiel dans la LMC des lymphocytes iNKT, cellules T de type « inné » auxquelles la littérature attribue de nombreuses fonctions antitumorales et des facteurs moléculaires, la cytokine/alarmine IL-33, produite dans la niche hématopoïétique, et son récepteur ST2 à la surface des cellules hématopoïétiques comme cibles de l’IL-33.La première partie de notre travail a ainsi permis de mettre en évidence de profondes altérations fonctionnelles des lymphocytes iNKT chez les patients atteints de LMC ainsi qu’une correction partielle de ces défauts après traitement par l’Imatinib (IM) ou l’IFN-. L’ensemble de ces résultats permet de proposer que l’altération des fonctions des cellules iNKT au cours du développement de la LMC pourrait participer aux mécanismes d’échappement de la tumeur au contrôle par le système immunitaire. La deuxième partie de notre travail a permis de mettre en évidence une expression de la molécule ST2, chaîne spécifique du récepteur à l’IL-33, à la surface des cellules CD34+ de patients atteints de LMC, expression non décelée chez les sujets sains et les patients en rémission après traitement par l’IM. De plus, contrairement aux cellules CD34+ de sujets sains, les cellules progénititrices de patients en phase chronique prolifèrent en réponse à l’IL-33. Enfin, nous avons montré que l’IL-33 est capable de contrecarrer in vitro les effets antiprolifératifs de l’IM. Ainsi nous pouvons émettre l’hypothèse selon laquelle l’IL-33, une cytokine/alarmine, puisse participer aux phénomènes conduisant à la persistance de progéniteurs hématopoïétiques leucémiques chez les patients sous traitement par IM. / To date, treatment of Chronic Myeloid Leukaemia (CML) is not sufficient to completely eradicate leukaemia cells. Hence, in order to develop a curative treatment, it is necessary to have a better understanding of the underlying mechanisms explaining why response to treatment is only partial..We therefore addressed the question whether the extrinsic mechanisms of CML control can affect the effectiveness of different treatments. We first provided evidence of profound functional impairments of iNKT cells in patients with CML. Interestingly these impairments were partially corrected after treatment with Imatinib (IM) or IFN-. Consequently our results suggest that altered functions of iNKT cells during the development of CML could facilitate tumour escape from immune destruction. . The second part of our work revealed that CD34+ progenitors from CML patients upregulate their cell surface expression of the IL-33-specific receptor chain ST2, proliferate and produce cytokines in response to IL-33, conversely to CD34+ cells from healthy individuals. Moreover, ST2 overexpression is normalized following IM therapy, while IL-33 counteracts in vitro IM-induced growth arrest in CML CD34+ progenitors. From these findings, it can be surmised that IL-33, a cytokine/alarmin likely expressed in the hematopoietic niche, facilitates the development of CML and IM resistance.

La protéine kinase D1, PKD1, est une nouvelle sérine/thréonine kinase activée par de nombreux mitogènes et dérégulée dans de nombreux types de cancers dont le cancer du sein, ce qui suggère un rôle de cette kinase dans la prolifération cellulaire et la tumorigenèse. Cependant, le rôle précis et les cibles de PKD1 ne sont pas encore bien connus. Au cours de ce travail, nous avons tout d’abord démontré que PKD1 est activée par les facteurs de croissance épidermique (EGF) et fibroblastique (FGF) et qu’elle régule la voie de signalisation de l’insulin-like Growth Factor-I (IGF-I). D’autre part, nos résultats démontrent que PKD1 favorise les propriétés pro-prolifératives et pro-tumorales des cellules MCF-7 dérivées d’un adénocarcinome mammaire humain estrogéno-dépendant. Ces mécanismes mettent en jeu des voies de signalisation dépendantes de protéines kinases (la voie MEK/ERK) et hormonales (la voie estrogène/REα). Ainsi, l’ensemble de ce travail fait apparaître PKD1 comme une nouvelle cible thérapeutique anti-tumorale potentielle. / Protein kinase D1, PKD1, is a novel serine/threonine kinase which can be activated by mitogens and whose expression is altered in many tumors such as breast cancer, suggesting a role for this kinase in cancer development. However, its precise role and targets are still unclear. Our study identified PKD1 as a new regulatory kinase implicated in the control of IGF-I signal transduction pathway. Furthermore, we showed that PKD1 enhances estrogen-dependent MCF-7 breast cancer cell proliferation and tumorigenesis through the regulation of MEK/ERK and estrogen/ERα pathways. Thus, this work may define PKD1 as a novel potential anti-tumor therapeutic target.

L’importance des activités protéolytiques associées à la membrane plasmique dans divers processus biologiques fondamentaux est de mieux en mieux définie. Les protéases de la famille ADAM (A Disintegrin and Metalloprotease), et ADAM10 en particulier, ont suscité un intérêt tout particulier du fait de l’importance de leurs substrats (récepteur de l’EGF, TNFα, Notch, APP…). Néanmoins, peu d’études se sont intéressées aux mécanismes régulant le trafic d’ADAM10.Les tétraspanines sont une super-famille de protéines de surface impliquées dans de nombreux processus biologiques fondamentaux parmi lesquels la migration, les interactions intercellulaires, la réponse immunitaire, la fusion des gamètes… L’une des caractéristiques majeure des tétraspanines est leur capacité à organiser un réseau d’interactions moléculaires appelé le « tetraspanin web ». De précédentes études menées dans le laboratoire ont montré qu’ADAM10 est associé au « tetraspanin web ». Néanmoins, la tétraspanine en interaction directe avec ADAM10 permettant son association au réseau n’est pas encore connue. Dans cette étude nous nous sommes intéressés à la régulation d’ADAM10 par les tétraspanines. Nous avons ainsi pu identifier une sous-famille de tétraspanines à 8 cystéines, les TspanC8 (Tspan5, Tspan10, Tspan14, Tspan15, Tspan17 et Tspan33), comme étant capables d’interagir directement avec ADAM10 et de réguler sa sortie du réticulum endoplasmique. Nous avons montré que Tspan5, Tspan14, Tspan15 et Tspan33 sont capables de réguler l’expression de surface d’ADAM10 et que Tspan10 et Tspan17 entrainent l’accumulation d’ADAM10 dans un compartiment endosomal tardif. Les TspanC8 pourraient également contribuer à la régulation de la spécificité de substrat d’ADAM10 puisque nous avons montré que l’expression des TspanC8 humaines Tspan5 et Tspan14 augmente l’activation de la voie Notch alors que Tspan15 n’a pas d’effet. Par ailleurs, les TspanC8 de Drosophile sont capables d’interagir directement avec Kuzbanian (l’orthologue d’ADAM10), permettent son accumulation à la surface cellulaire et régulent l’activation de la voie Notch dans différents contextes développementaux. Nous proposons que les TspanC8 soient une nouvelle famille de protéines ayant une fonction très conservée dans la régulation de l’activité et du trafic d’ADAM10, capables de réguler l’activation de la voie Notch. / Increasing evidence suggests a critical implication of membrane-associated protease activities in numerous biological processes. ADAM (A Disintegrin and Metalloprotease) proteases, and especially ADAM10, are of particular interest because of the importance of their substrates (EGF receptor, TNF α, Notch, APP…). However, few studies focus on the mechanisms of ADAM10 trafficking. Tetraspanins are a super-family of proteins implicated in numerous biological processes including migration, intercellular interactions, immune response, gamete fusion… One of the most striking features of tetraspanins is their ability to organise multi-molecular complexes called « Tetraspanin Web ». Previous studies in the laboratory have shown that ADAM10 is associated to the « Tetraspanin Web ». Nevertheless, the tetraspanin in direct interaction with ADAM10 that drives its association to the web is not known. In this study, we focused on ADAM10 regulation by tetraspanins. We identified a subfamily of tetraspanins with 8 cysteines in their large extracellular domain that we called TspanC8 (Tspan5, Tspan10, Tspan14, Tspan15, Tspan17 and Tspan33) that can directly interact with ADAM10 and regulate its egress from the endoplasmic reticulum. We have shown that Tspan5, Tspan14, Tspan15 and Tspan33 regulate the surface expression of ADAM10 and that Tspan10 and Tspan17 accumulate ADAM10 in a late endosomal compartment. TspanC8 could also contribute to substrate specificity since Tspan5 and Tspan14 can increase Notch activation when Tspan15 cannot. Drosophila TspanC8 directly interact with the Drosophila ADAM10 ortholog Kuzbanian, increase its accumulation at the cell surface and modulate Notch activation in several developmental contexts. We propose that TspanC8 constitute a new family of Notch regulators with conserved functions in the regulation of ADAM10 trafficking and activity.

Ce travail de thèse a pour objectif la simplification de la préparation d’un nouveau type d’inhibiteur d’entrée du VIH conçu, synthétisé et validé dans le cadre d’une collaboration entre l’équipe de Glycochimie Moléculaire et Macromoléculaire dont je dépends, l’Institut de Biologie Structurale de Grenoble et l’Institut Pasteur de Paris. Ce prototype est constitué d’un mime fonctionnel de CD4 lié de façon covalente à un fragment dodécasaccharidique d’Héparane Sulfate dont la synthèse est complexe. Nous avons donc proposé de préparer des oligomaltosides sulfatés afin de déterminer s’ils pouvaient se comporter comme des mimes d’Héparane Sulfate.Dans un premier temps, nous avons mis au point la synthèse, en huit étapes et 38 % de rendement global, d’un précurseur trisaccharidique oligomérisable à partir de maltotriose, un trisaccharide biosourcé commercial. Au cours de ce travail, nous avons résolu trois points particulièrement délicats : l’allylation de l’extrémité réductrice du maltotriose, l’installation d’un groupement paraméthoxybenzylidène en position O-4III et O-6III et la protection sélective des positions O-6I et O-6II par un groupement silylé. Les optimisations menées nous ont permis de limiter la formation de produits secondaires, d’augmenter le rendement de chaque étape et de pouvoir mener sans problème cette synthèse sur une échelle d’une dizaine de grammes. Dans un deuxième temps, le précurseur trisaccharidique a été transformé en différents accepteurs et donneurs de glycosyle dont les comportements dans différentes conditions de glycosylation ont été étudiés. Nous avons ainsi pu démontrer qu’une activation des donneurs sous forme de trichloroacetimidate conduisait à des rendements faibles de part la formation d’une quantité importante des produits de réarrangement en trichloroacétamides anomériques. Une activation sous forme de N-Phényltrifluroacétimidate a permis de résoudre ce problème, sans toutefois que les rendements en soient toujours augmentés. En effet, nous avons pu montrer que la nature du groupement protecteur en O-6I du donneur a une influence déterminante sur l’issue de la réaction de glycosylation, tant au niveau de sa stéréosélectivité que de son rendement. Un groupement encombré ou un ester en O-6I du donneur est ainsi indispensable pour avoir une bonne stéréosélectivité alpha. Le meilleur rendement obtenu est, pour le moment, de 56 %. Des optimisations en cours permettront d’augmenter le rendement et de préparer les oligomaltosides sulfatés visés dans un avenir proche afin de tester leur activité biologique. / This work aims at simplifying the preparation of a new type of HIV entry inhibitor, conceived, synthesized and validated within a collaboration between our group, the "Institut de Biologie Structurale" (Grenoble) and the Institut Pasteur (Paris). This prototype is composed of a CD4 functional mimetic linked to a dodecasaccharide fragment of Heparan Sulfate, whose synthesis is complex. In order to determine if Heparan Sulfate may be replaced by simpler sulfated oligosaccharides, we decided to prepare a set of sulfated oligomaltosides.To this goal, we first optimized the synthesis of an oligomerizable maltotrioside building block in eight steps and 38% global yield from maltotriose, a commercial and biosourced trisaccharide. In this work, we had to address three major points: the allylation of the reducing end of maltotriose, the introduction of a paramethoxybenzylidene group between positions O-4III and O-6III and the selective protection of the remaining primary positions O-6I and O-6II by a silylated protecting group. Each step has been optimized to minimize the amount of secondary products and thus to enhance its yield. The resulting synthesis was thus shown to be highly reproducible up to ten grams scale.Then, glycoside acceptors and donors were prepared from the oligomerizable maltotrioside building block and we studied their behaviors in glycosylation reactions. We found that trichloroacetimidate activation led to poor glycosylation yields, due to the competitive formation of trichloroacetamidyl glycoside rearrangement product. Gratifyingly, N-phenyltrifluroacetimidate activation solved the rearrangement problem, but yields sometimes remained low. Indeed, we were able to demonstrate that the nature of the protecting group in position O-6I of the donor strongly influenced both the stereoselectivities and yields of the glycosylations: a bulky or ester group is needed in this position to obtain a full alpha stereoselecticity. To date, the highest yield obtained is 56 %.Ongoing optimizations will allow us to enhance the yields and to prepare the targeted sulfated oligomaltosides in a near future in order to test their biological activity.

Submitted by Beatriz Aline Riga null (beatriz_aline16@hotmail.com) on 2017-03-15T18:07:31Z No. of bitstreams: 1 Riga Rocha, B. A._me_sjrp.pdf: 4397938 bytes, checksum: 97cd69af09e5df0e1dcc372a541946c2 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2017-03-21T16:17:50Z (GMT) No. of bitstreams: 1 rocha_bar_me_sjrp.pdf: 4397938 bytes, checksum: 97cd69af09e5df0e1dcc372a541946c2 (MD5) / Made available in DSpace on 2017-03-21T16:17:50Z (GMT). No. of bitstreams: 1 rocha_bar_me_sjrp.pdf: 4397938 bytes, checksum: 97cd69af09e5df0e1dcc372a541946c2 (MD5) Previous issue date: 2017-02-22 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Neste trabalho objetivou-se a síntese e caracterização de oito complexos de cobalto(II) que fossem aptos a atuarem como agentes controladores na Polimerização Radicalar Mediada por Cobalto do monômero acetato de vinila. Através da modulação da esfera de coordenação destes complexos com a alteração dos efeitos estéricos e eletrônicos promovidos pela coordenação de ligantes α-diiminas, buscou-se estabelecer parâmetros para propor a forma como estes complexos mediam a CMRP do referido monômero. Os complexos foram divididos em dois grupos, objetivando uma avaliação mais eficiente da forma como os efeitos estéricos e eletrônicos interferem na capacidade controladora dos mesmos. Após a variação sistemática das proporções molares entre complexo, iniciador e monômero encontrou-se na proporção [Co]/[AIBN]/[VAc] = 1/3,25/542 os resultados mais satisfatórios deste trabalho. O uso de DMSO fora prejudicial para os complexos 2a-c, porém fez com que os complexos 2d-h apresentassem melhorias, reduzindo seus valores de polidispersidade e aumentando as massas moleculares dos polímeros. O complexo 2b fora escolhido como o melhor controlador do grupo do efeito estérico, por reunir em si as características estérias e eletrônicas que o fizeram apresentar os menores valores de polidispersidade (Đmáx = 1,43). Com relação ao grupo de complexos do efeito eletrônico, fora o complexo 2f que, devido à maior estabilidade estrutural conferida ao complexo por seu substituinte cicloexil, apresentara as menores polidispersidades de seu grupo, entre 1,12 e 1,58. / This work aimed the synthesis and characterization of eight cobalt(II) complexes which could be able to act as controlling agents in vinyl acetate radical polymerization mediated by cobalt. By modulating the coordination sphere of these complexes, modifying the steric and electronic effects offered by the α-diimine ligands, we sought to establish parameters to propose how these complexes mediate the CMRP of vinyl acetate. The complexes were split in two groups in way to assess efficiently how the electronic and steric effects can affect the controlling capacity of the CoII(α-diimine).A systematic variation of the molar ratio such as [Co]/[initiator] and [Co]/[monomer] made it possible to choose [Co]/[initiator]/[monomer] = 1/3.25/542 as the ratio that provided certain level of control. The addition of DMSO was detrimental to the complexes 2a-c, but caused an improvement to the activity of complexes 2dh, reducing their polydispersity values and increasing the molecular weights of the polymers. Complex 2b was chosen as the best controller of the steric effect group, its steric and electronic characteristics made possible to reach the lowest polydispersity of its group (Đmax = 1.43).Regarding to the group of complexes of the electronic effect, it was the complex 2f that had the smallest polydispersity of its group, between 1.12 and 1.58, due to the greater structural stability conferred to the complex by its cyclohexyl substituent.

MyD88 est une protéine adaptatrice clé dans la signalisation des TLRs/IL-1R qui mène à l'activation de NF-KB et des MAPK, et à la production de cytokines inflammatoires. MyD88 participe à la tumorigénèse par le biais de son activité inflammatoire dans la signalisation des TLRs/IL-1R, et également via son interaction directe avec la kinase Erk dans la cellule cancéreuse. Dans cette thèse, nous identifions de nouveaux partenaires protéiques de MyD88 et nous examinons comment leurs interactions peuvent réguler sa fonction. Nous démontrons que MyD88 interagit avec Ubc9, ce qui conduit à sa sumoylation, et que cette modification posttraductionnelle régule négativement l'inflammation dépendante de MyD88. Nos résultats montrent également que MyD88 interagit avec le récepteur nucléaire, ER-α, et que cette interaction est nécessaire pour la réponse inflammatoire. Enfin, nous avons étudié l'importance de l'interaction MyD88/Erk dans le maintien de la transformation des tumeurs dépendant de l'oncogène Ras. Ces résultats pourraient éventuellement être exploités pour cibler MyD88 et ses interactions dans le traitement des maladies inflammatoires et le cancer / MyD88 is a protein that is at the interface between inflammation and cancer. It is the key adaptor protein used by TLRs/IL-1R to mediate their downstream signaling, resulting in NF-κB and MAPK activation, and inflammatory cytokine production. MyD88 also plays a role in tumorigenesis via two mechanisms, an inflammatory one dependent on its function in TLRs/IL- 1R signaling, and an intrinsic, cell-autonomous mechanism mediated by its interaction with the kinase Erk. Based on the different roles played by MyD88, this thesis work consisted in studying how MyD88 protein-protein interactions can regulate its function. We show that MyD88 interacts with Ubc9, resulting in its sumoylation and subsequent negative regulation of MyD88- mediated inflammation. We also demonstrate that MyD88 interacts with the nuclear receptor ER-α, an interaction necessary for the inflammatory response. Finally, we have studied the importance of the MyD88/Erk interaction in the maintenance of the transformed phenotype of Ras-dependent tumors. These findings could eventually be exploited to target MyD88 and its interactions in the treatment of inflammatory disorders and cancer

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No. of bitstreams: 1 PDF - Antônio Manoel da Silva Filho.pdf: 1626282 bytes, checksum: 35f4f3bebdefb1abb1e520955b76a0e8 (MD5) Previous issue date: 2016-02-18 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The utilization of Gluconacetobacter diazotrophicus in agriculture has shown promise in optimization morphophysiological aspects, biochemical and yield of plants. The culture of red rice (Oryza sativa L.) presents great socioeconomic and environmental importance in the semi- arid northeast. The production of this culture still suffers by the not full use of appropriate technologies, due to scarcity of studies and development of technologies. Given the above, the objective of this study was to evaluate the potential effect of G. diazotrophicus on germination and initial growth of red rice.The experiment was conducted in a completely randomized design, with six treatments (SNE-seeds not soaked in water, SE H2O-seeds soaked in water for 24 hours, SE H2O + GA3 seed soaked in gibberellic acid solution for 24 hours, SE H2O + GD-seeds soaked in water for 24 + G. diazotrophicus, SE H2O + GA3 + GD-seeds soaked in gibberellic acid solution for 24 hours + G. diazotrophicus and SNE + GD-seeds not soaked in water + G. diazotrophicus) with six replicates.The concentration of gibberellic acid (GA3) used was 50 mg -1 L . Evaluated were the aspects physiological, biochemical, molecular and initial growth of red rice seedlings. Data were submitted to analysis of variance, mean test and Pearson correlation. Treatments significant effect on the growth variables, physiological, biochemical and molecular.It was verified that treatment with non-imbibed seeds inoculated with G. diazotrophicus promote increase of 28.7; 41.7; 28.7; 49.5; 69.6; 48.5; 61.5; 38.5; 46; 97; 86 and 89% for the variables: germination, germination speed index, first count, root length, shoot length, total fresh mass, total dry mass of α-amylase activity, activ acid phosphatase, expression GAMYB transcription factor, α-amylase and S-adenosyl-L-methionine (SAM) synthase, respectively, in relation to the treatment of seeds not soaked in water and not inoculated.It was concluded that the inoculation with G. diazotrophicus red rice seeds enhances the speed of germination and seedling germination, root length, shoot length, and acid phosphatase activities of α-amylase, total fresh mass, total dry mass, GAMYB expression of the transcription factor, α- amylase expression and SAM. Therefore presents great potential agronomic and biotechnology for use as growth promoter in the red rice crop, increasing the germination and early growth independently of seed imbibition. / A utilização de Gluconacetobacter diazotrophicus na agricultura tem-se mostrado promissora na otimização de aspectos morfofisiológicos, bioquímicos e rendimento das plantas. Acultura do arroz vermelho(Oryza sativa L.) apresenta grande importância socioeconômica e ambiental no semiárido nordestino. Aprodução dessa cultura ainda padece pela não utilização plena de tecnologias apropriadas, devido a escassez de estudos e desenvolvimento de tecnologias. Diante do exposto, objetivou-se com este trabalho avaliar o efeito potencial da G. diazotrophicussobre a germinação e crescimento inicialde arroz vermelho. O experimento foi realizado emdelineamento inteiramente casualisado, constando de seis tratamentos (SNE-sementes não embebidas em água, SE H2O-sementes embebidas em água por 24h, SE H2O + GA3-sementes embebidas em solução de ácido giberélicopor 24h, SE H2O + GD-sementes embebidas em água por 24h + G. diazotrophicus, SE H2O + GA3 +GD-sementes embebidas em solução de ácido giberélicopor 24h + G. diazotrophicuse SNE + GD-sementes não embebidas em água + G. diazotrophicus), com seis repetições. A concentração da solução de ácido giberélico (GA3) usada -1 foi de 50 mg L . Avaliaram-se os aspectos fisiológicos, bioquímicos, moleculares e de crescimento inicial das plântulas de arroz vermelho. Os dados foram submetidos à análise de variância, teste de médias e correlação de Pearson. Os tratamentos exerceram efeito significativo sobre as variáveis de crescimento, fisiológicas,bioquímicas e moleculares. Registrou-se que o tratamento com sementes não embebidas e inoculadas com G. diazotrophicuspromoveram incrementos de 28,7; 41,7; 28,7; 49,5; 69,6; 48,5; 61,5; 38,5; 46; 97; 86 e 89% para as variáveis: germinação, índice de velocidade de germinação, primeira contagem de germinação, comprimento radicular, comprimento da parte aérea, massa fresca total, massa seca total, atividade da α-amilase, ativida da fosfatase ácida, expressão do fator de transcrição GAMYB, α- amilase e S-adenosil-L-metionina (SAM) sintetase, respectivamente, em relação ao tratamento das sementes não embebidas em água e não inoculadas. Concluiu-se que a inoculação de G. diazotrophicus em sementes de arroz vermelho aumenta a velocidade de germinação e germinação das plântulas, comprimento radicular, comprimento da parte aérea, atividade s da fosfatase ácida e α-amilase, massa fresca total, massa seca total, a expressão do fator de transcrição GAMYB, expressão de α-amilase e SAM.Portanto apresenta grande potencial agronômico e biotecnológico para aplicação como promotora de crescimento na cultura do arroz vermelho, incrementando a germinação e crescimento inicial de modo independente de embebição das sementes.

Submitted by Daniella Santos (daniella.santos@ufma.br) on 2017-08-29T16:11:39Z No. of bitstreams: 1 MayaraSilva.pdf: 2720807 bytes, checksum: 1a7eeabdd7df89c4b7c0690e8136cb51 (MD5) / Made available in DSpace on 2017-08-29T16:11:39Z (GMT). No. of bitstreams: 1 MayaraSilva.pdf: 2720807 bytes, checksum: 1a7eeabdd7df89c4b7c0690e8136cb51 (MD5) Previous issue date: 2017-03-28 / CNPq / CAPES / FAPEMA / There is an increasing interest to find new products with therapeutic potential to the treatment of leishmaniasis, due the high toxicity and resistance of the majority of available treatments. Our aim was to formulate, characterise and evaluate the antiLeishmania amazonensis activity of babassu loaded poly(lactic-co-glycolic acid) – PLGA microparticles. The PLGA microparticles were loaded with the aqueous extract of babassu mesocarp (MMP) and evaluated for size, zeta potential, drug content, encapsulation efficiency in comparison to unloaded microparticles (CMP). The antiLeishmania effect was evaluated to promastigotes forms or to amastigotes in Balb/c macrophage cells infected with Leishmania amazonensis. Following macrophage treatment with MMP the percent of infected cells was determined by Giemsa staining and compared with cells treated with CMP or with free babassu extract (MESO). To find the potential mechanisms of the activity of MMPs, TNF -α, IL-6, IL-10, hydrogen peroxide, arginase and accumulated nitrite in the culture supernatants of the treated and untreated cells were also measured by ELISA, and by colorimetric assays, respectivelly. The size range of the microparticles was between 3 and 6,4 μm with an average zeta potential of −25 mV and encapsulation efficiency of 45%. The antiLeishmania activity of babassu-loaded microparticles was 10-fold higher than MESO. MMP showed an overall bioavailability and hence were more effective in eliminating intracellular parasites than the other formulations. Babassu microparticles also reduced the ex vivo parasite infectivity and this effect seems to be directly related to a polarization of macrophages to the M1 phenotype with an increased production of nitric oxide, hydrogen peroxide and TNF-α. Interestingly, this overexpression of TNF-α didn’t impair cell viability, suggesting the anti-apoptotic effects of the MMP in infected macrophages. These findings indicate that babassu load microparticles may be useful for targeting for new drugs, due to the immunomodulatory effects of polarization to M1 macrophages, infected with L. amazonensis, and further provide motivations for future studies on human cels in vitro and in animal models of leishmaniasis in vivo. / A bioprospecção de produtos com potencial terapêutico no tratamento da leishmaniose tem despertado crescente interesse, pois as drogas atualmente utilizadas apresentam elevada toxicidade e, muitas vezes, os protozoários são resistentes, sobretudo nos tratamentos prolongados. Na perspectiva de desenvolver uma nova formulação com ação anti-Leishmania avaliou-se a atividade do extrato aquoso do mesocarpo de babaçu (Attalea speciosa Mart) encapsulado em micropartículas biodegradáveis de PLGA [poly(lactic-co-glycolic acid]. Inicialmente, foi realizado o estudo morfométrico e funcional das micropartículas. Em seguida foi avaliada a atividade anti-Leishmania por ação sobre as formas promastigotas, comparando os efeitos das micropartículas de PLGA carregadas com extrato do mesocarpo de babaçu (MMP) com o extrato livre (Meso) e com micropartículas sem o mesocarpo, utilizadas como controle negativo (CMP). Também avaliamos os efeitos de MMP em culturas de macrófagos peritoneais, de camundongos Balb/c, infectados ou não com formas amastigotas de Leishmania amazonensis. Os seguintes parâmetros foram investigados nos sobrenadantes das culturas de macrófagos: quantificação das citocinas IL-10, IL-6 e TNF-α por ELISA, detecção de peróxido de hidrogênio, óxido nítrico e atividade da arginase. Foi determinada a taxa de infecção e o percentual de células infectadas. O mesocarpo de babaçu apresentou forteinteração com antígenos de L. amazonensis. A caracterização das micropartículas mostrou que as MMP apresentaram diâmetro e potencial zeta compatíveis com as micropartículas controle (CMP) e a eficiencia de encapsulamento do extrato foi de 45%. As MMPs foram mais ativamente fagocitadas do que o extrato de babaçu livre, ocasionando aumento de 25% no índice fagocítico, após 24 horas de incubação. Além de baixa toxicidade para macrófagos peritoneais, o encapsulamento do mesocarpo de babaçu potenciou em quase 10 vezes a ação anti-Leishmania para as formas promastigotas de Leishmania amazonensis, quando comparado ao extrato livre. O tratamento com MMP reduziu o número de amastigotas e a taxa de infecção de macrófagos peritoneais, possivelmente por seu efeito imunomodulador na polarização de macrófagos para o fenótipo M1, resultando no aumento de TNF-α e óxido nítrico e na inibição da produção de IL-10. Concluímos que o microencapsulamento do mesocarpo de babaçu melhorou a ação anti-Leishmania do extrato, mas manteve o seu efeito imunomodulador o que contribuiu para o melhor efeito tanto sobre os protozoários como para as células infectadas, evitando a morte das células por necrose ou apoptose. Os dados em conjunto indicam que as micropartículas MMP podem ser fortes candidatas ao desenvolvimento de novos produtos, devido aos seus efeitos imunomoduladores na polarização de macrófagos infectados com L. amazonensis para um perfil M1 e, adicionalmente, estimulam novos estudos quanto ao seus efeitos sobre células humanas in vitro e em modelo animal da leishmaniose in vivo.

<p>Autoimmune mechanisms and genetic susceptibility contribute to the pathogenesis of primary Sjögren’s syndrome and SLE. These chronic systemic autoimmune diseases have many serological and clinical features in common and have an impact on daily life. The studies in this thesis aim to elucidate their autoimmune mechanisms, define susceptibility genes and evaluate effects of androgen supplement on health-related quality of life.</p><p>Autoantibodies against α-fodrin, a widely distributed cytoskeletal protein, were detected at similar frequencies in sera from patients with primary and secondary Sjögren’s syndrome and SLE. Consequently, testing for antibodies against α-fodrin would not add diagnostic value compared to conventional serological analysis and does not discriminate between these diseases.</p><p>The type I interferon (IFN) system was found to be activated in primary Sjögren’s syndrome. IFN-α containing cells were detected in minor salivary gland biopsies, while sera from patients with primary Sjögren’s syndrome induced IFN-α production in the presence of apoptotic and necrotic cell material. This ability of sera correlated with the presence of antibodies against RNA-binding proteins and IFN-α production was dependent on RNA in immune complexes. The natural interferon producing cells/plasmacytoid dendritic cells (NIPC/PDC) were the IFN-α producers and blocking of FcγRIIa inhibited the production. Single nucleotide polymorphisms (SNPs) in two genes in the type I IFN signalling pathway, those for tyrosine kinase 2 and interferon regulatory factor 5, were strongly associated with SLE in a Swedish, Finnish and Icelandic population. The minor allele frequencies were lower in SLE patients than in healthy controls. These SNPs may decrease the function of the type I IFN system, thereby conferring protection against SLE. </p><p>Supplementation with dehydroepiandrosterone (DHEA) in glucocorticoid treated women with SLE led to mild improvements in health-related quality of life in respect of mental well-being and sexuality, whereas physical well-being was unaffected.</p>