Partie 1 : De plus en plus de molécules fluorées sont utilisées dans bon nombre de domaines variés, allant des matériaux aux sciences de la vie. Ces dernières années, un intérêt croissant a émergé avec l'association du groupement CF3 avec un hétéroatome, comme OCF3 ou SCF3. Le groupement SCF3 est très intéressant à cause de son paramètre d'hydrophobie (π=1.44). Par conséquent, les composés portant ce groupement sont des cibles importantes pour de nombreuses applications, en particulier en chimie médicinale. Cependant, la majorité des précédentes méthodes décrites dans la littérature utilisent des réactifs toxiques dans des conditions drastiques. Les trifluorométhanesulfénamides (1ère et 2nde génération) ont démontré leur potentiel dans la trifluorométhylthiolation électrophile. En raison de leur réactivité intéressante, ces deux générations de réactifs stables sont maintenant dans la boîte à outils de la chimie organique pour la trifluorométhylthiolation de molécules. Partie 2 : Des aggrégats d'α-synucléine sont une caractéristique neuropathologique de nombreuses maladies neurodégénératives, notamment la maladie de Parkinson (MP) et la démence à corps de Lewy (DLB), collectivement appelés synucléinopathies. L'imagerie TEP pourrait révéler la quantité et la distribution des agrégats d'α-synucléine dans le cerveau et serait plus avantageuse à utiliser pour le diagnostic spécifique de synucléinopathies présymptomatiques à différents stades de lamaladie. Nous avons concentré nos efforts sur des dérivés de benzimidazole comme composés de petites tailles, plans et π-délocalisés pour concevoir des traceurs radioactifs des agrégats de la synucléine. Ainsi des structures assemblant des benzimidazoles, un espaceur rigide (alcyne et triazole) et enfin une autre partie aromatique ont été envisagées. Le radiomarquage pourra être effectué par une substitution nucléophile avec K18F au cours de la dernière étape. Avec cette stratégie convergente, nous pourrions avoir accès à une grande série de molécules à évaluer / Part 1 : More and more applications for fluorinated molecules are being found in various fields, from materials to life sciences. In recent years, a growing interest has emerged in the association of the trifluoromethyl group with heteroatoms such as CF3O or CF3S. The CF3S moiety is of particular interest, because of its high hydrophobicity parameter (π=1.44). Consequently compounds bearing this group are important targets for various applications, in particular in medicinal chemistry and agrochemistry. However, the majority of previous methods described in the literature use toxic reagents under harsh conditions. Trifluoromethanesulfenamides (1st and 2nd generation) have demonstrated their potential in the electrophilic trifluoromethylthiolations. Because of their interesting reactivity, these two generations of shelf-stable reagents are now in the toolbox of organic chemists for the trifluoromethylthiolation of molecules, providing a convenient method to pursue less toxic pathways. Part 2 : α-synuclein aggregation is a neuropathological hallmark of many neurodegenerative diseases including Parkinson’s disease (PD) and dementia with Lewy bodies (DLB), collectively termed synucleinopathies. PET imaging can reflect the amount and distribution of alpha-synuclein aggregates in the brain and would be advantageous to use for specific diagnosis of synucleinopathies in presymptomatic stages of disease. We focused our interest onto benzimidazole derivatives as small, planar and -delocalized compounds to design radiotracers of synuclein aggregates. Compounds based on the association of benzimidazole moiety, rigid linker (alkyne and triazole) and another aromatic part have been designed. The radiolabeling could be performed by nucleophilic substitution with K18F during the last step. With this convergent strategy, we could have acces to a large series of molecules to be evaluated

<p>Bikunin, a chondroitin sulfate-containing protein of 25 kDa, has protease inhibitory activity and occurs in the plasma in free and complexed form. In inter-α-inhibitor (IαI) and pre-a-inhibitor (PαI) it is covalently linked through its chondroitin sulfate (CS) chain to two or one other polypeptide of about 80 kDa – heavy chains 1 and 2 (H1, H2) and heavy chain 3 (H3) – respectively. Bikunin and the heavy chains are synthesized as precursors, which are proteolytically cleaved and assembled into IαI and PαI in the secretory pathway. The C-terminal extension (CTX) of the heavy chains seems to mediate its own cleavage and theassembly of the complexes. The heavy chains of the IαI family become transferred to hyaluronan during ovulation and inflammation.</p><p>In this thesis, the biosynthesis of PαI, the plasma clearance of bikunin and the binding of IαI to collagen were studied. We found that in H3, a short segment on the N-terminal side of the CTX cleavage site is required for cleavage. Furthermore, the H3 could become linked to free CS chains primed by a xyloside, showing that the bikunin protein core is not needed for coupling. We also identified His649 as a residue essential for coupling, but not for cleavage. </p><p>Bikunin labelled with a residualizing agent, 125I-tyramine cellobiose, was injected into mice to identify tissues involved in its uptake. Half of the radioactivity was recovered in the kidneys, 10% in the liver, and the rest distributed in other tissues. We determined the half-life of bikunin in rat plasma using two independent methods: injection of 125I-bikunin, or hepatectomy followed by assessing the rate of disappearance of endogenous bikunin. Both methods yielded half-time values of 5-7 minutes. Removal of the CS chain did not affect the clearance rate of bikunin.</p><p>IαI and its heavy chains were found to bind to collagen with dissociation constants greater than 2 μM and 0.4-0.6 μM, respectively and this binding was independent of divalent metal ions. We suggest that the interaction of IαI with collagen may play a modulatory role in cell migration or in remodelling of the extracellular matrix.</p>

Bikunin, a chondroitin sulfate-containing protein of 25 kDa, has protease inhibitory activity and occurs in the plasma in free and complexed form. In inter-α-inhibitor (IαI) and pre-a-inhibitor (PαI) it is covalently linked through its chondroitin sulfate (CS) chain to two or one other polypeptide of about 80 kDa – heavy chains 1 and 2 (H1, H2) and heavy chain 3 (H3) – respectively. Bikunin and the heavy chains are synthesized as precursors, which are proteolytically cleaved and assembled into IαI and PαI in the secretory pathway. The C-terminal extension (CTX) of the heavy chains seems to mediate its own cleavage and theassembly of the complexes. The heavy chains of the IαI family become transferred to hyaluronan during ovulation and inflammation. In this thesis, the biosynthesis of PαI, the plasma clearance of bikunin and the binding of IαI to collagen were studied. We found that in H3, a short segment on the N-terminal side of the CTX cleavage site is required for cleavage. Furthermore, the H3 could become linked to free CS chains primed by a xyloside, showing that the bikunin protein core is not needed for coupling. We also identified His649 as a residue essential for coupling, but not for cleavage. Bikunin labelled with a residualizing agent, 125I-tyramine cellobiose, was injected into mice to identify tissues involved in its uptake. Half of the radioactivity was recovered in the kidneys, 10% in the liver, and the rest distributed in other tissues. We determined the half-life of bikunin in rat plasma using two independent methods: injection of 125I-bikunin, or hepatectomy followed by assessing the rate of disappearance of endogenous bikunin. Both methods yielded half-time values of 5-7 minutes. Removal of the CS chain did not affect the clearance rate of bikunin. IαI and its heavy chains were found to bind to collagen with dissociation constants greater than 2 μM and 0.4-0.6 μM, respectively and this binding was independent of divalent metal ions. We suggest that the interaction of IαI with collagen may play a modulatory role in cell migration or in remodelling of the extracellular matrix.

In Anlehnung an die Definition beim Pferd (SCHOON et al. 1992, 1997) wird die bovine Endometrose als endometriale periglanduläre und/oder stromale Fibrose mit Alteration der betroffenen Drüsen definiert (RODENBUSCH 2011). Eine eingehende histomorphologische und immunhistologische Charakterisierung der Endometrose existiert bisher bei der Stute (KENNEY u. DOIG 1986, SCHOON et al. 1992, HOFFMANN et al. 2009), jedoch nicht beim Rind. Das Ziel dieser Arbeit ist daher die histomorphologische und immunhistologische Charakterisierung der verschiedenen Endometroseformen beim Rind. Die Auswertung der Proben erfolgt am Institut für Veterinär-Pathologie der Universität Leipzig. In Abhängigkeit von den klinisch-gynäkologischen Befunden werden diese Proben in drei Gruppen unterteilt. Gruppe A1: Endometriumbioptate (n=12) von vier klinisch-gynäkologisch gesunden fertilen Rindern in definierten Zyklusphasen; Gruppe A2: Endometriumbioptate (n=36) von 36 klinisch genitalgesunden Rindern mit mindestens einer Abkalbung und Gruppe B: Uterusquerschnitte (n=69) von 69 sub-/ infertilen Rindern. Die Proben werden anhand der von HOFFMANN (2006) definierten Kriterien auf histopathologische Veränderungen hin untersucht und charakterisiert. Das histomorphologische Erscheinungsbild der involvierten periglandulären Stromazellen erlaubt die Einteilung der Endometrose in eine aktive, inaktive und gemischte Fibrose, die, je nach der Integrität des Drüsenepithels, einen destruierenden oder nicht destruierenden Charakter aufweist. Zur Charakterisierung der Endometroseformen werden neben histomorphologischen Kriterien die Intermediärfilamente Desmin, Vimentin und Zytokeratin sowie die Expression von α-Aktin und Laminin berücksichtigt. Die deskriptive statistische sowie die Inferenzstatistik-Auswertung erfolgen unter Zuhilfenahme der Software SPSS 18. Eine aktive Fibrose tritt bei 96,2 % der Rinder auf; 1,9 % weisen eine inaktive und 1,9 % eine gemischte Endometrose auf. Ein nicht destruierendes Erscheinungsbild der Endometrose kann bei 88,6 % der untersuchten Rinder nachgewiesen werden. Bei 11,4 % der untersuchten Rinder liegt eine Endometrose mit destruierendem Charakter vor. Die Endometrose betrifft bei 81 % der Rinder Einzeldrüsen und bei 19 % Drüsennester. Drüsennester sind häufiger bei mittel- und hochgradigen Endometrosen nachweisbar. 20 % der Proben mit nicht destruierender Endometrose und 58 % der Proben mit destruierender Endometrose weisen eine entzündliche Infiltration der Drüsen auf, wobei ein Zusammenhang zwischen der entzündlichen Infiltration der endometrotischen Drüsen und dem destruierenden Charakter der Endometrose festgestellt werden kann. Zusätzlich zur Endometrose zeigen 61 % der Rinder eine Endometritis, 12,4 % eine Perivaskulitis und 66 % eine interkarunkuläre und/oder intrakarunkuläre Angiosklerose. Insgesamt findet sich nur bei 24 % der Fälle ausschließlich eine Endometrose, bei 10,5 % eine Endometrose und zugleich eine Endometritis, bei 16 % liegt eine Endometrose zusammen mit einer Angiosklerose vor. Eine Kombination von Endometrose, Angiosklerose und Endometritis ist bei 49,5 % der Proben nachweisbar. Insgesamt bestehen jedoch keine erkennbaren statistischen Zusammenhänge zwischen den einzelnen Befundkombinationen. Aufgrund der immunhistologischen Untersuchung kann konstatiert werden, dass die periglandulären Stromazellen innerhalb der Endometrose eine stromale Koexpression von Desmin, Vimentin und α-Aktin aufweisen, welche ein für Myofibroblasten charakteristisches Merkmal ist. Ein kleiner Prozentsatz der Drüsenepithelzellen in der destruierenden Endometrose reagiert multifokal positiv mit dem Vimentinantikörper. Dies ist möglicherweise Ausdruck einer Fehldifferenzierung zur Stabilisierung der Zelle oder Anzeichen einer intensivierten (pathologischen) Proliferation. Die Lamininexpression der Basallamina der endometrotisch veränderten Drüsen ist, insbesondere bei der destruierenden Endometrose, diskontinuierlich und geht mit einer Auffaserung der Basallamina einher. Vermutlich lassen sich die umfangreichen Basallaminaalterationen auf von Myofibroblasten sezernierte Enzyme zurückführen. Bei Rindern kann kein Zusammenhang zwischen der Endometrose und dem Alter der Rinder oder der Anzahl der Kalbungen festgestellt werden. In der vorliegenden Arbeit dominiert die geringgradig aktive nicht destruierende Endometrose gegenüber den anderen bovinen Endometroseformen. Die sub-/ infertilen Kühe (Gruppe B) zeigen häufiger eine schwerere und destruierende Endometrose als die klinisch gesunden Rinder (Gruppe A1, A2). Die klinisch-gynäkologisch gesunden Rinder in definierten Zyklusphasen weisen variable Endometroseformen oder Endometrosegrade auf. Die sub-/ infertilen Rinder zeigen eine höhere Güstzeit (252,82 ± 163,83 Tage) als die klinisch-gynäkologisch gesunden Tiere mit mindestens einer Abkalbung (94 ± 28,4 Tage). Die längere Güstzeit bei den sub- und infertilen Rindern könnte somit eine Folge des insgesamt in Charakter und Grad stärker geschädigten Endometriums bei dieser Gruppe sein. Somit kann unter Berücksichtigung der vorliegenden Ergebnisse angenommen werden, dass die aufgeführten Alterationen die Fertilität des Rindes negativ beeinflussen. Die Ergebnisse ermöglichen eine histomorphologische und immunhistologische Charakterisierung der Endometrose beim Rind. Anhand der Ergebnisse der hier durchgeführten detaillierten Untersuchungen ist es möglich, eine präzisere Deskription degenerativer endometrialer Befunde vorzunehmen. In Hinblick auf die Fertilität bei Vorliegen einer bovinen Endometrose wird somit die Grundlage für zukünftige prognostische Bewertungen gelegt.

De ekologiskt betydelsefulla småbiotoperna blir allt färre i det moderna jordbruket på grund av intensifieringen av jordbrukslandskapet som resulterat i ett allt mer fragmenterat landskap. För att kunna ge småbiotoper, såsom åkerholmar, ett optimalt skydd behövs flera studier om organismerna på holmarna, deras diversitet och spridning. I denna undersökning studeras artrikedomen av mossor och lavar på busk- och trädbekädda samt öppna åkerholmar i ett öppet- och ett skogsdominerat landskap. Analysen av artrikedom i studien tyder på att de finns fler arter av mossor och lavar i det öppna landskapet i förhållande till det skogsdominerade landskapet samt på de åkerholmarna som saknade busk- och trädvegetation. Dessa miljöer består av torr och stenig mark och växer igen relativt långsamt, vilket är gynnsamt för vissa mossor och lavar. Det fanns ingen signifikant korrelation mellan diversitet och storleken på åkerholmarna eller mellan diversitet och holmarnas avstånd till skog. Antalet arter i ett habitat kan bero på regionala artpoolen samt antalet potentiella kolonisatörer men även lokala processer. Vidare är det viktigt att nämna att studien tyder på att diversitet av mossor och lavar påverkas av förhållandena på holmarna, såsom busk- och trädtäckningen. Detta i kombination med en variation inom dessa små biotoper kan skapa goda förutsättningar för ett landskap med hög diversitet. / The ecologically important small remnant habitats are becoming fewer in modern agriculture due to the intensification of agricultural landscapes that have resulted in an increasingly fragmented landscape. In order to provide an ideal conservation framework for small habitats, such as midfield islets, several studies about the organisms on the islets, their diversity and distribution are needed. In this study I investigated the diversity of mosses and lichens on open midfield islets with and without shrubs and trees in an open and a forest-dominated landscape. The analysis of species diversity in the study suggests that, overall; there are more species of mosses and lichens in the open landscape in relation to the forest-dominated landscape, and on the opened islets lacking shrub and tree vegetation. These environments consist of dry and rocky soils and overgrow relatively slowly, which is favourable for mosses and lichens. There was no significant correlation between diversity and size of the midfield islets or their distance to forest. The number of species in a habitat may depend on regional species pool and the number of potential colonizers but also local processes. Furthermore, it is important to mention that the study suggests that the diversity of mosses and lichens is influenced by the conditions on the midfield islands, such as shrub and tree cover. These conditions, combined with a variation within these small habitats can create good conditions for a landscape with high diversity.

Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2012-07-19T19:54:59Z No. of bitstreams: 1 Gisele Batista Carvalho. Análise da expressão dos Genes e IGF-1, HGF, VEGF - CPqGM - Dissertação de Mestrado - 2008.pdf: 634399 bytes, checksum: c4a7c8d54e5c114e87ff5e2c2cd711b0 (MD5) / Made available in DSpace on 2012-07-19T19:54:59Z (GMT). No. of bitstreams: 1 Gisele Batista Carvalho. Análise da expressão dos Genes e IGF-1, HGF, VEGF - CPqGM - Dissertação de Mestrado - 2008.pdf: 634399 bytes, checksum: c4a7c8d54e5c114e87ff5e2c2cd711b0 (MD5) Previous issue date: 2008 / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, Brasil / Nos últimos anos estudos têm demonstrado a participação de citocinas e fatores de proliferação e diferenciação na regeneração do miocárdio em condições de agressão. Os fatores que influenciam a regeneração do miocárdio na fase aguda da infecção por T. cruzi na qual ocorre um processo massivo de destruição do miocárdio, ainda são desconhecidos. Neste trabalho foi investigada a expressão dos genes de HGF, IGF-1, VEGF e TNF-α no coração de camundongos durante o curso da infecção aguda por T. cruzi. Fragmentos de corações de camundongos C57Bl/6 infectados com cepa Colombiana de T. cruzi, sacrificados com 15, 25 30, 40 e 60 dias pós-infecção, foram analisados pela técnica de PCR em tempo real. A expressão dos genes de HGF, IGF-1, VEGF e TNF-α no coração foi avaliada e correlacionada ao grau de inflamação, fibrose e parasitemia nos tempos de infecção analisados. Foi observado um aumento significativo na expressão dos genes de TNF-α e IGF-1 durante toda a fase aguda da infecção, enquanto que a expressão de HGF apresentou-se diminuída. Já a expressão do gene de VEGF não foi significativamente alterada no decorrer da infecção. As expressões dos genes de HGF, IGF-1 e TNF-α apresentaram uma correlação negativa com a parasitemia e a inflamação. Estes resultados indicam que o miocárdio produz vários mediadores solúveis em resposta à infecção por T. cruzi, que podem ter uma participação tanto nos mecanismos de lesão, como o TNF-α, como na recuperação da lesão tecidual promovida pela infecção por T. cruzi, como é o caso do IGF-1. Outros estudos devem ser realizados no sentido de melhor entender o papel destes mediadores produzidos localmente ou trazidos pela circulação no processo de reparo do miocárdio pela infecção por T. cruzi. / Studies carried out in the past few years have demonstrated the participation of cytokines and growth factors related to cell proliferation and differentiation in the regeneration of the myocardium after injury. The factors involved in the regeneration of the myocardium during the acute phase of T. cruzi infection, when a process of massive destruction of the myocardium occurs, are not known. In this work we investigated the gene expression of HGF, IGF-1, VEGF e TNF-α in the hearts of mice during the acute infection by T. cruzi. Heart fragments of C57Bl/6 mice infected by Colombian strain T. cruzi were obtained 15, 25 30, 40 e 60 days after infection and analyzed by real time PCR. The gene expression of HGF, IGF-1, VEGF, and TNF-α in the heart was evaluated and correlated to the degree of inflammation, fibrosis and parasitemia at the time points analyzed. A significant increase in TNF-α and IGF-1 gene expression was observed during the acute phase of infection, whereas the expression of HGF was decreased. The expression of VEGF gene was not significantly altered during infection. The expression of HGF, IGF-1, and TNF-α genes showed a negative correlation with parasitemia and inflammation. The results indicate the miocardium as a source of soluble mediators, in response to T. cruzi infection, which may participate both in the mechanisms of lesion induction, in the case of TNF-α, as well as in the repair of tissue lesions promoted by T. cruzi infection, such as IGF-1. Additional studies should be carried out in order to understand the role of these mediators produced locally or brought by the circulation in the process of lesion repair in the miocardium during T. cruzi infection.

Resumo: Este trabalho objetivou avaliar a influência da vitamina E pela suplementação por meio da dieta, em peixes submetidos a dois métodos de abate (imersão em água gelada e sangria), nos atributos de qualidade de filés congelados de tilápia (Oreochromis spp.). Peixes, com peso médio inicial de 340 g, foram alimentados por um período de nove semanas com três dietas isocalóricas (3264,09 kcal ED/kg) e isoprotéicas (24,8% PD), diferindo na adição de 100 mg/kg de ração de α-tocoferol (grupo TO), 100 mg de princípio ativo/kg de ração de acetato de α- tocoferil (grupo AC), e sem adição de vitamina E (grupo controle - CO). Ao final do período de alimentação, os peixes foram abatidos pelos métodos citados anteriormente. Utilizou-se delineamento inteiramente casualizado, em esquema fatorial 3x2x5, caracterizado por três dietas, dois métodos de abate e cinco tempos de análises dos filés. Foram realizadas análises de desempenho e corporais, e de composição centesimal, oxidação lipídica pela formação de substâncias reativas ao ácido tiobarbitúrico (SRATB), e sensorial pelo método do Índice de Qualidade (MIQ) nos filés, nos tempos zero (antes do congelamento), 45, 90, 120 e 150 dias de estocagem. A suplementação da vitamina E não afetou os parâmetros de desempenho, entretanto influenciou na composição centesimal e na oxidação lipídica, protegendo os filés ao longo do período de estocagem. Foi elaborada tabela de avaliação dos filés congelados, pelo MIQ, instrumento útil para a análise de atributos de qualidade. / Abstract: The aim of this investigation was to evaluate the influence of vitamin E dietary supplementation, and fish slaughtering methods (immersion in ice-water and exsanguination), on the fillets quality of tilapia, during frozen storage. Fish of 340 g mean initial body weight were fed for nine weeks with three isoenergetic (containing 3264,09 kcal DE/kg) and isonitrogenous (24,8% DP) diets. Two diets were supplemented with 100 mg/kg diet of α-tocopherol (TO group) and 100 mg of active source/kg diet of α-tocopheryl acetate (AC group), plus a nonsupplemented diet (control group - CO). At the termination of the 9-week feeding trial, fish were slaughtered by the summoned methods.A completely randomized design was used, in a 3x2x5 factorial scheme, characterized by the 3 vitamin E supplementation diets, 2 slaughter methods and 5 fillets analysis. The growth performance parameters, hepatosomatic and fat viscerosomatic index, centesimal composition, lipid oxidation determined by the thiobarbituric acid reactive substances (TBARS) and sensorial by Quality Index Method (QIM) analysis were analyzed in five times: before frozen storage, 45, 90, 120 and 150 days storage. The vitamin E supplementation did not influence the growth performance parameters, but centesimal composition were influenced, and vitamin E protected fillets from lipid oxidation in frozen storage. Fillets quality availability table were organized, by QIM, for to help in sensorial analysis. / Orientadora: Léa Silvia Sant'Ana / Coorientador: João Batista Kochenborger Fernandes / Banca: Cristiane Rodrigues Pinheiro Neiva / Banca: Cláudio Angelo Agostinho / Mestre

Submitted by JÚLIO HEBER SILVA (julioheber@yahoo.com.br) on 2018-07-09T18:58:27Z No. of bitstreams: 2 Dissertação - Ana Paula Lopes Lima - 2018.pdf: 2335802 bytes, checksum: 2f6ce764d8628ef3ceadc17fda527b72 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-07-10T11:27:45Z (GMT) No. of bitstreams: 2 Dissertação - Ana Paula Lopes Lima - 2018.pdf: 2335802 bytes, checksum: 2f6ce764d8628ef3ceadc17fda527b72 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-07-10T11:27:45Z (GMT). No. of bitstreams: 2 Dissertação - Ana Paula Lopes Lima - 2018.pdf: 2335802 bytes, checksum: 2f6ce764d8628ef3ceadc17fda527b72 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-06-22 / Leishmaniasis is a disease transmitted by protozoan-infected sandflies of the genus Leishmania, whose cutaneous form is the most common with 50 to 75% of new cases reported. It is a non-contagious disease of chronic evolution, considered stigmatizing and neglected, affecting about 500,000 people in the world. Most drugs used in pharmacological treatment are potentially toxic and require patient hospitalization for administration, which raises costs and discourages the needy population from adhering to treatment. In this scenario, herbal medicines are inserted as an alternative because of their proven efficacy and because they cause fewer undesirable effects. The objective of the present study was to evaluate the leishmanicidal potential and the in vitro cell viability of Siparuna guianensis (SG) and α- Bisabolol (AB) isolated essential oils of Siparuna guianensis (SG) on Vero cells and Leishmania amazonensis promastigotes using the method colorimetric resazurin salt (Alamar Blue®) as an indicator of cellular metabolism. Cell invasion was also performed with the parasites previously treated with the IC50 obtained for AB and SG obtained in the cell viability test, in order to test the effect of the compounds on the infection capacity of the parasites. The results were analyzed using the program GraphPad Prism version 6.01. For analysis of the IC50, the non-linear regression tests were used. The tested compounds demonstrated viability on Vero basal cells and leishmanicidal activity: SG IC50 131.5 μg/mL and 2.59 μg/mL respectively. For α-Bisabolol the concentrations obtained were 103.8 μg/mL and 6.394 μg/mL and for SG were 94.61 μg/mL and 7.84 μg/mL, respectively. This is the first report of a cell invasion assay made with S. guianensis and α-Bisabolol alone, in which inhibition of 98.4% infective capacity of L. amazonensis promastigotes treated with SG at 131,5 μg/mL and inhibition of 95.4% of the infective capacity of the same AB-treated parasite at a concentration of 95.4 μg/mL, which are close to the value obtained by the conventional drug in a similar in vitro assay. which suggests a refinement of the technique and in vivo assays to determine the action of S. guianensis and α-Bisabolol from the concentrations obtained in this research. / A leishmaniose é uma doença transmitida por insetos do tipo flebotomíneos infectados por protozoários do gênero Leishmania, cuja forma cutânea é a mais comum com 50 a 75% de casos novos relatados. É uma doença não contagiosa, de evolução crônica, considerada estigmatizante e negligenciada, afetando cerca de 500.000 pessoas no mundo. A maioria das drogas utilizadas no tratamento farmacológico é potencialmente tóxica e requer hospitalização do paciente para sua administração, o que eleva os custos e desestimula a população carente a aderir ao tratamento. Neste cenário, os medicamentos fitoterápicos inserem-se como uma alternativa devido a sua eficácia comprovada e por causarem menos efeitos indesejáveis. O objetivo do presente estudo foi avaliar o potencial leishmanicida e a viabilidade celular in vitro do óleo essencial de Siparuna guianensis (SG) e do α-Bisabolol (AB) isolado de S. guianensis sobre células Vero e promastigotas de Leishmania amazonensis, utilizando o método colorimétrico sal de resazurina (Alamar Blue®) como indicador de metabolismo celular. Foi realizado também ensaio de invasão celular com os parasitos previamente tratados com o IC50 obtido para AB e SG obtido no teste de viabilidade celular, afim de testar o efeito dos compostos na capacidade de infecção dos parasitos. Os resultados foram analisados utilizando o programa GraphPad Prism versão 6.01. Para análise do IC50 foi utilizado os testes de regressão não linear. Os compostos testados demonstraram viabilidade sobre células basais Vero e atividade leishmanicida: SG IC50 131,5 µg/mL e 2,59 µg/mL respectivamente. Para α-Bisabolol as concentrações obtidas foram 103,8 µg/mL e 6,394 µg/mL e para SG foram 94,61 µg/mL e 7,84 µg/mL, respectivamente. Este é o primeiro relato de um ensaio de invasão celular feito com S. guianensis e α-Bisabolol isolado, no qual pode ser observado a inibição de 98,4% de capacidade infectiva de promastigotas de L. amazonensis tratadas com SG na concentração de 131,5 µg/mL e inibição de 95,4 % da capacidade infectiva do mesmo parasito tratado com AB na concentração de 95,4 µg/mL, resultados estes que se aproximam do valor obtido pelo medicamento convencional em um ensaio semelhante in vitro, o que sugere um refinamento da técnica e ensaios in vivo para determinar a ação da S. guianensis e do α-Bisabolol a partir das concentrações obtidas nesta pesquisa.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2018-01-29T11:11:37Z No. of bitstreams: 2 Tese -Gustavo Lage Costa - 2014.pdf: 4404595 bytes, checksum: 914021b79a6b38b0b2af9d94eebc4c97 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-01-29T11:13:18Z (GMT) No. of bitstreams: 2 Tese -Gustavo Lage Costa - 2014.pdf: 4404595 bytes, checksum: 914021b79a6b38b0b2af9d94eebc4c97 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-01-29T11:13:18Z (GMT). No. of bitstreams: 2 Tese -Gustavo Lage Costa - 2014.pdf: 4404595 bytes, checksum: 914021b79a6b38b0b2af9d94eebc4c97 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2014-12-22 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The grass Brachiaria sp. brought to Brazil in the green revolution, came with the goal of increasing the Brazilian livestock production. The introducing of the grass, starts emerging problems such as leaf hopper pastures and hepatogenous photosensitization. The etiology of photosensitization caused by Brachiaria sp. is still controversial. But not even knowing for sure rightly if the cause is the steroidal saponins or the fungus Phytomices chartarium, it is known that they occur and cause damage to livestock. The described laboratory abnormalities are increased activity of enzymes aspartate aminotransferase (AST) and gamma glutamyl transferase (GGT), bilerrubinemia and jaundice, and evidence of insufficient functionof.liver. Histopathological examination can demonstrate a cholangiohepatitis and presence of foamy macrophages. The presence of these cells is important because its quantification was negatively correlated with the gain in cattle weight. Thus this study evaluated 40 Nelore young bulls, with the Brachiaria sp hay consumption adjusted to the maximum amount of forage (84%), with the goal of reproducing the hepatic abnormalities described in cases of photosensitization. Therefore, the aim of this study was to evaluate the possible liver changes, by the metabolic and histological profile of the liver and lymph nodes, and also check if the offered antioxidants would be able to minimize these changes, promoting greater weight gain and improving carcass characteristics of feedlot cattle. As a result, the braquiaria hay with low amounts of saponin and fungal spores did not induce clinical signs of liver damage in young bulls, but laboratory assessments indicate evidence of chronic inflammatory liver change with the presence of foamy macrophages. The antioxidant supplementation did not alter the metabolic profile of the animals, but reduced the amount of foamy macrophages in the liver. However this decrease did not improve weight gain of cattle, carcass characteristics and the final quality of the meat. / O capim Brachiaria sp. trazido para o Brasil na revolução verde, veio com o objetivo de aumentar a produção pecuária brasileira. Juntamete com o capim, começararam a surgir problemas, como a cigarrinha das pastagens e a fotossenssibilização hepatógena. A etiologia da fotossenssibilização causada pelo capim Brachiaria sp. ainda é controversa. Mas mesmo não sabendo ao certo se o causador é as saponinas esteroidais ou o fungo Phytomices chartarium, sabese que elas ocorrem, e causam prejuízos à pecuária. As alterações laboratoriais descritas são o aumento de atividade das enzimas aspartato aminotransferase (AST) e gama glutamiltransferase (GGT), bilerrubinemia e icterícia, além de indícios de insuficiência da função hepática. Os exames histopatológicos podem demonstrar uma colangiohepatite e presença de macrófagos espumosos. A presença destas células é importante, pois a sua quantificação foi correlacionada negativamente com o ganho em peso de bovinos. Desta forma, o objetivo deste estudo foi avaliar 40 tourinhos nelore, com o consumo de feno de Brachiaria sp. ajustado para a maior quantidade possível de volumoso (84%), no intuto de reproduzir as alterações hepáticas descritas em casos de fotossensibilização hepatógena, que foram monitoradas por meio de testes como o perfil metabólico e histológico do fígado e linfonodos, e ainda verificar se os antioxidantes ofertados seriam capazes de minimizar estas alterações, promovendo maior desenvolvimento ponderal e melhorando as características da carcaça de bovinos confinados. Como resultados, o feno de braquiaria com baixas quantidades de saponina e de esporos do fungo, não induziram sinais clínicos de lesão hepática nos tourinhos. Porém as avaliações laboratoriais indicam evidencias de alteração hepática inflamatória crônica com presença de macrófagos espumosos. A suplementação com antioxidantes não alterou o perfil metabólico dos animais, mas reduziu a quantidade dos macrófagos espumosos no fígado. Essa diminuição não melhorou o desenvolvimento ponderal dos bovinos, as características de carcaça e a qualidade final da carne.

Submitted by Marilene Donadel (marilene.donadel@unioeste.br) on 2018-06-04T23:33:22Z No. of bitstreams: 1 Elissandro_Klein_2016.pdf: 4516260 bytes, checksum: 605fbf2e69fd746b1152b1431bf0dca5 (MD5) / Made available in DSpace on 2018-06-04T23:33:22Z (GMT). No. of bitstreams: 1 Elissandro_Klein_2016.pdf: 4516260 bytes, checksum: 605fbf2e69fd746b1152b1431bf0dca5 (MD5) Previous issue date: 2016-02-04 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Eugenia pyriformis Cambess. (Uvaia), typical of the Atlantic Forest belongs to the Myrtaceae family, the same family of cherry, jabuticaba and other plants, currently studied due to their antioxidant, antimicrobial and medicinal properties. The interest in bioactive compounds obtained from natural sources has led to an increase in researches aiming to find these compounds in plant extracts. The extractions using conventional methods generally have some drawbacks, and an interesting alternative has been the use of non-conventional extraction technologies, such as the supercritical fluid and the ultrasound-assisted extraction. This work aims to study unconventional methods of obtaining uvaia extracts in order to obtain extracts rich in bioactive compounds. Factors such as the influence of type of extraction (supercritical fluid and ultrasound) and the used conditions on the yield and composition were evaluated. The results were compared to those obtained using the extraction by maceration, a conventional extraction method. After drying and milling the uvaia leaves, the plant material was subjected to extraction with the supercritical CO2 solvent under different conditions of temperature (40, 50 and 60 °C) and pressure (100, 150 and 200 bar), with a fixed flow of 2.0 kg min-1. In the ultrasound-assisted extraction, the effects of temperature (40, 50 and 60 °C), power (150, 250 and 350 W) and ratio between the mass of leaves and the volume of solvent (1:10, 1:15 and 1:20) were evaluated. The extracts obtained were chemically characterized by GC-MS giving β-amyrin (53.72 % for SFE and 71.25 % for UAE) and α-amyrin (24.63 % for SFE and 22.69 % for UAE) as main compounds. In the supercritical extraction, the pressure and the interaction between pressure and temperature showed statistically significant effects on the yield, with the best result being achieved at 200 bar and 60 °C, reaching 1.69%. The data obtained from the supercritical fluid extraction were used to adjust the extraction curves simulated by using two empirical models available in the literature. The tested models adjusted well to the experimental data and the analysis of the estimated parameters allowed to define which model best describes the kinetics of each experimental condition. For the ultrasound-assisted extraction, the best yield result found in the determination of the kinetics was 1.81% in 40 minutes, although in the experimental design a 3 minute time extraction was used due to the better characteristics showed by the extracts at this time. The best result obtained in experimental design with ultrasound was 1.79% provided 30% power, 60 °C and mas/solvent ratio 1:20. Statistical analysis of the experimental design of the ultrasound-assisted extraction indicated significant effects from the variables temperature, mass/solvent ratio and the interaction between power and mass/solvent ratio. The best results for amyrin content by extract weight was found for supercritical extraction, with 97.43%. The ultrasound-assisted extraction showed the best amyrin amount per mass of leaves, 12.13 g of the mixture of isomers per kg of dry leaf. / Eugenia pyriformis Cambess. (Uvaia), típica da Mata Atlântica pertence à família Myrtaceae, mesma família da pitanga, jabuticaba e outras plantas, atualmente estudadas devido às suas propriedades antioxidantes, antimicrobianas e medicinais. O interesse por compostos bioativos obtidos de fontes naturais tem levado ao aumento no número de pesquisas que visam encontrar esses compostos em extratos de plantas. As extrações que utilizam os métodos convencionais geralmente apresentam inconvenientes, e uma alternativa interessante tem sido a utilização de tecnologias não convencionais de extração, como a extração utilizando fluidos supercríticos e a extração assistida por ultrassom. O presente trabalho tem como objetivo estudar métodos não convencionais de obtenção de extratos de uvaia visando obter-se extratos ricos em compostos bioativos. Foram avaliados fatores como influência do tipo de extração (fluido supercrítico e ultrassom) e das condições utilizadas no rendimento e na composição. Os resultados foram comparados com os obtidos utilizando a extração por maceração, um método convencional de extração. Após secagem e moagem das folhas de uvaia, o material vegetal foi submetido à extração com o solvente CO2 supercrítico sob diferentes condições de temperatura (40, 50 e 60 oC) e de pressão (100,150 e 200 bar), com vazão fixa de 2,0x10-3 kg min-1. Na extração assistida por ultrassom, foram avaliados os efeitos da temperatura (40, 50 e 60 oC), da potência (150, 250 e 350 W) e razão entre massa de folhas e volume de solvente (1:10, 1:15 e 1:20). Os extratos obtidos foram caracterizados quimicamente por CG-EM apresentando com principais compostos β-amirina (53,72 % para EFS e 71,25 % para EAU) e α-amirina (24,63 % para EFS e 22,69 % para EAU). Na extração supercrítica, a pressão e a interação entre pressão e temperatura apresentaram efeitos estatisticamente significativos em relação ao rendimento, sendo que o melhor resultado foi encontrado com 200 bar e 60 oC, obtendo-se 1,69 %. Os dados obtidos na extração com fluido supercrítico foram utilizados para ajustar as curvas de extração simuladas utilizando dois modelos empíricos disponíveis na literatura. Os modelos testados ajustaram-se de forma satisfatória aos dados experimentais e a análise dos parâmetros estimados permitiu definir qual dos modelos melhor se ajustou à cinética de cada condição experimental. Para a extração assistida por ultrassom o melhor resultado de rendimento encontrado na determinação da cinética foi 1,81 % com 40 minutos, porém no planejamento experimental foi utilizado um tempo de extração de 3 minutos pois com esse tempo os extratos apresentavam melhores características, sendo portanto o melhor resultado obtido igual a 1,79 % na condição de 30 % de potência, 60 oC e razão massa/solvente de 1:20. A análise estatística do planejamento experimental da extração assistida por ultrassom indicou efeitos significativos das variáveis temperatura, razão massa/solvente e a interação entre potência e razão massa/solvente. Os melhores resultados de teor de amirina por massa de extrato foi encontrada para a extração supercrítica, com 97,43 %. A extração assistida por ultrassom apresentou a melhor quantidade de amirina por massa de folha, 12,13 g da mistura de isômeros por kg de folha seca.