Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-05-23T14:10:14Z No. of bitstreams: 1 2015_dis_mspereira.pdf: 700705 bytes, checksum: 72d54e714f0c7b084f064aea398bf72e (MD5) / Approved for entry into archive by Nádja Goes (nmoraissoares@gmail.com) on 2016-05-23T14:10:52Z (GMT) No. of bitstreams: 1 2015_dis_mspereira.pdf: 700705 bytes, checksum: 72d54e714f0c7b084f064aea398bf72e (MD5) / Made available in DSpace on 2016-05-23T14:10:52Z (GMT). No. of bitstreams: 1 2015_dis_mspereira.pdf: 700705 bytes, checksum: 72d54e714f0c7b084f064aea398bf72e (MD5) Previous issue date: 2015 / PEREIRA, Magnum de Sousa. Avaliação das taxas de mineralização do carbono e nitrogênio do composto orgânico proveniente de carcaça e despojo de pequenos ruminantes. 2015. 48 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Centro de Ciências Agrárias, Departamento de Ciências do Solo - Programa de Pós-Graduação em Agronomia - Solos e Nutrição de Pantas, Fortaleza-CE, 2015 / The determination of the mineralization rate of organic compost is necessary for planning the most efficient way to use them. Thus, this study aimed to determine the carbon and nitrogen mineralization rate from organic compost produced from sheep and goat carcasses and its slaughtering spoils. Chromic Inceptisol (Luvisols) samples were incubated at an average temperature of 30.5 ° C with doses of 0; 3.75; 7.5; 15 and 30 Mg ha-1 of organic compost. To evaluate the carbon mineralization were used 100 g of soil incubated with these doses arranged in a completely randomized design (CRD) distributed in a split plot scheme. Samples were kept in glass containers tightly closed and the C-CO2 measurements were performed during periods of 0; 1; 2; 3; 4; 5; 6; 7; 9; 11; 14; 17; 20; 23; 26; 29; 33; 37; 41; 48; 55; 69; 83; 97; 112 and 126 days after the start of incubation. To determine the nitrogen mineralization rate (N), the doses of compost were incubated with 100 g of soil and distributed in a CRD with a 5 x 10 factorial arrangement. The assessment of inorganic N were performed at 7; 14; 28; 42; 56; 70; 84; 98; 112 and 126 days after the incubation beginning. Both models, the simple exponential and the double exponential, were not efficient to explain the dynamics of C mineralization for not consider the interactions that occurs when the compost is applied to the soil. A model that considers the soil C labile and recalcitrant compartments (ls and rs), protected and unprotected compartments of the applied organic matter (OM) (pc and dc) and a p factor that modifies the rate of mineralization of soil organic matter (SOM) when the compost is applied (C0 = Cls e-kltp + Crs.e-krstp + Cpc .e-kpst + Coc .e-kdct) was more efficient to explain the dynamics of C, considering the interactions with the SOM and the OM added. The suggested model has demonstrated that the rate of SOM decomposition is approximately 10% greater in the presence of the compost and the compost mineralization rate is 0.012 day-1, explaining the 97.95% of the variability in the data. The N mineralization was very fast since 40% of the standard dose of 7.5 Mg h-1 was found in the mineral form 14 days after the incubation. However, due to losses of inorganic nitrogen by NH3 volatilization, it was not possible to estimate the actual N mineralization rate. / A determinação da taxa de mineralização de compostos orgânicos se faz necessária para o planejamento da forma mais eficiente de sua utilização. Deste modo, objetivou-se determinar a taxa de mineralização de carbono e nitrogênio de composto orgânico produzido a partir de carcaças e despojos de abate de ovinos e caprinos. Amostras de Luvissolo Crômico foram incubadas à temperatura média de 30,5 ºC com doses equivalentes a 0; 3,75; 7,5; 15 e 30 Mg ha-1 de composto orgânico. Para avaliação da mineralização do carbono foram utilizados 100 g de solo incubados com as referidas doses dispostas em um delineamento inteiramente casualisado (DIC) distribuídos em esquema de parcela subdividida. As amostras foram mantidas em recipientes de vidro hermeticamente fechados sendo as mensurações de C-CO2 realizadas nos períodos de 0; 1; 2; 3; 4; 5; 6; 7; 9; 11; 14; 17; 20; 23; 26; 29; 33; 37; 41; 48; 55; 69; 83; 97; 112 e 126 dias após o início da incubação. Para a determinação da taxa de mineralização do nitrogênio (N), as doses de composto foram incubadas com 100 g de solo e distribuídas em um DIC dispostas em um arranjo fatorial de 5 x 10. As avaliações do N inorgânicoforam realizadas aos7; 14; 28; 42; 56; 70; 84; 98; 112 e 126 dias após o início da incubação. Tanto o modelo simples exponencial quanto o modelo duplo exponencial não foram eficientes para explicar a dinâmica de mineralização do C por não considerarem as interações que ocorrem quando o composto é aplicado ao solo. Um modelo que considera os compartimentos de C lábil e recalcitrante no solo (ls e rs), compartimentos protegido e desprotegidos da MO aplicada (pc e dc) e um fator p que modifica a taxa de mineralização da MO do solo quando o composto é aplicado (C0 = Cls e-kltp + Crs.e-krstp + Cpc .e-kpst + Cdc .e-kdct) se mostrou mais eficiente para explicar a dinâmica do C, considerando as interações da MO do solo com a MO adicionada. O modelo sugerido demonstrou que a taxa de decomposição da MO do solo é aproximadamente 10% maior na presença do composto e a taxa de mineralização do composto é de 0,012 dia-1, explicando 97,95% da variabilidade dos dados. A mineralização do N mostrou-se bastante rápida visto que 40% da dose padrão de 7,5 Mg ha-1 foi encontrado na forma mineral 14 dias após a incubação. No entanto, devido às perdas de nitrogênio inorgânico por volatilização de NH3, não foi possível se estimar as reais taxas de mineralização do N.

Ces travaux centrés essentiellement sur la fonction thiohydroximate s’articulent selon trois thèmes : (1) les acides thiohydroxamiques, (2) la synthèse et (3) l’étude de réactivité des N-oxydes de thioimidate (TINOs), avec pour objectif l’étude d’une enzyme : la S-glucosyltransférase (S-UGT). La S-UGT est une enzyme clé dans la formation des glucosinolates. Les travaux menés ont tout d’abord été axés sur la synthèse d’acides thiohydroxamiques, substrats de la S-UGT, pour l’étude cinétique de l’enzyme. Dans ce but, nous avons envisagé différentes méthodes et avons pu obtenir les molécules ciblées, permettant ainsi d’obtenir des résultats préliminaires quant à la cinétique de l’enzyme. D’autre part, la synthèse d’iminosucres, inhibiteurs potentiels de l’enzyme, a été envisagée par une double fonctionnalisation d’un N-oxyde de thioimidate (TINO) sur charpente pyranose. Nous avons donc développé une méthode robuste d’accès aux TINOs à partir de différents pyranoses. Le couplage de Liebeskind-Srogl a permis d’accéder aux nitrones. Des additions-éliminations sur les TINOs nous ont donné accès à des N-oxydes d’amidine et à de nouveaux TINOs. Les réactions de réduction et de débenzylation des TINOs ont également été explorées. Des tentatives de cycloaddition 1,3-dipolaire sur une nitrone ont été effectués en vue d’obtenir à des isoxazolidines, précurseurs d’iminosucres Alpha, Alpha-disubstitués. Les résultats concernant la synthèse d’inhibiteurs de la S-UGT restent préliminaires. Toutefois, après déprotection, les différentes molécules synthétisées durant cette thèse pourront être envisagées comme inhibiteurs de glycosidases. / This thesis is articulating into three topics: (1) synthesis of thiohydroxamic acids, (2) the synthesis and (3) the study of the reactivity of thioimidate N-oxides (TINOs). The purpose of these works is the study of the enzyme: S-glycosyltransferase (S-UGT). The enzyme S-UGT is an essential enzyme for the glucosinolate formation.The starting point of these works was focused on the synthesis of thiohydroxamic acids. These products are the substrates of the enzyme S-UGT and could be used for the study of the enzyme kinetic. The second point is to use TINOs anchored on pyranose backbones as precursors of iminosugars which are potential inhibitors of the enzyme S-UGT. An efficient methodology was developed to access a large panel of TINOs from different pyranoses. Liebeskind Srogl coupling allowed us to obtain nitrones. Addition-elimination on TINOs gave us amidine N-oxides and new TINOs. Reduction and debenzylation reactions on TINOs were also explored. 1,3-Dipolar cycloaddition attempts on a nitrone were performed to obtain isoxazolidines which are precursors Alpha, Alpha-disubstituted iminosugars. The results recording the synthesis of S-UGT inhibitors remain preliminary. However, after deprotection, the different molecules, synthesised during this thesis, will be tested as glycosidase inhibitors.

Made available in DSpace on 2014-06-11T19:33:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-08-14Bitstream added on 2014-06-13T20:05:40Z : No. of bitstreams: 1 silva_ce_dr_botfmvz.pdf: 254829 bytes, checksum: 7c94d454e44648c813b0838e17fe1709 (MD5) / Universidade Estadual Paulista (UNESP) / O objetivo deste trabalho foi comparar em pastagem de capim-tobiatã (Panicum maximum cv. Tobiatã) o efeito da aplicação de duas fontes de nitrogênio, convencional (uréia) e alternativa (Amiorgan®) em relação a produtividade da pastagem e eficiência de utilização. O experimento foi realizado de janeiro a dezembro de 2008. Utilizou-se delineamento inteiramente casualizado composto por 2 fontes nitrogenadas, uréia e Amiorgan®, com 8 repetições cada. A dose de N utilizada somente no período das águas foi de 50 kg de N/ha/ciclo de pastejo. Produtividade da parte aérea da pastagem e extração e recuperação de nitrogênio pela mesma foram semelhantes entre tratamentos. Como os teores foliares de N estão acima dos recomendados na literatura e a extração e recuperação de N pela planta foram relativamente baixas, sugere-se que efeitos residuais de adubações anteriores tenham influenciado a resposta da pastagem ao estímulo da adubação nitrogenada. / This paper aims the evaluation of productivity and efficiency of an alternative nitrogen fertilizer (Amiorgan®) compared with conventional fertilizer in Panicum maximum cv Tobiatã pasture. The experiment was conducted from January to December, 2008. The treatments used two different nitrogen fertilizers: urea and Amiorgan®. The statistical application used was randomized blocks with eight repetitions of each. In the rain season (summer), the nitrogen dosage used was 50 Kg of N/ha/grazing cycle. Area pasture productivity, extraction and recovery of the nitrogen were similar among treatments. As the levels of N found on the leaves is above those recommended in the literature and the extraction and recovery of N by the plant were relatively low, it is suggested that residual effects of previous fertilizations have influenced the response of pasture to the stimulus of nitrogen fertilization.

O presente trabalho pretende analisar as fugas das Sonatinas nº3 e nº6 para piano de Camargo Guarnieri, através de uma análise estilística e tendo em vista os procedimentos utilizados pelo compositor especificamente no tratamento da fuga como técnica composicional. Guarnieri compôs 8 sonatinas para piano escritas entre 1928 e 1982, período que abrange grande parte da vida criativa do compositor. A análise das duas as fugas enfocará cinco parâmetros, conforme o esquema analítico de Jan LaRue: Som, Harmonia, Melodia, Ritmo e Crescimento. Através desses parâmetros, verificaremos como se desenvolvem estruturalmente as fugas, destacando os aspectos mais importantes que caracterizarão o discurso musical desenvolvido pelo compositor.

La Prodigiosine est un métabolite secondaire de couleur rouge produit principalement par la bactérie Serratia marcescens. Cette molécule, de structure tripyrrolique, présente des activités antitumorales, immunosuppressives, antibiotiques, antifongiques et antipaludiques. Ses nombreuses activités en font un composé potentiellement intéressant pour des applications thérapeutiques. Notre équipe a développé une méthodologie de synthèse basée sur la réactivité de dérivés pyrroliques vis-à-vis des nitrones et des N-hydroxylamines pour mener sélectivement à des structures dipyrrométhanes et dipyrrométhènes. Cette méthodologie a été le point de départ de la première étude de ce manuscrit. Nous avons ainsi développé une voie d'accès vers la synthèse d'analogues de la Prodigiosine. Ces derniers, substitués en position méso, n'ont jamais fais l'objet d'étude. Pour cela, il a fallu développer une voie d'accès au 4-méthoxy-2,2'-bipyrrole, précurseur principal de la Prodigiosine. Nous avons également mis au point une voie d'accès à des 2,2'-bipyrroles dissymétriques. Ces derniers sont des briques élémentaires utilisés en chimie des porphyrines étendues. Ces macrocycles pyrroliques trouvent des applications dans de nombreux domaines allant des polymères conducteurs aux cristaux liquides en passant par des applications biologiques. Notre méthodologie permet la formation de bipyrroles en six étapes, à partir de substrats facilement accessibles, avec des rendements globaux pouvant atteindre 20 %. Cette méthode a également été étendue à la synthèse du motif terpyrrole. / Prodigiosin is a secondary metabolite mainly produced by Serratia marcescens. This red pigment possessing a tripyrrolic structure, shows antitumor, immunosuppressive, antibiotic, antifungal and anti-malarial activities. These activities make it a valuable compound for therapeutic applications. Our team has developed a synthetic methodology based on the addition of pyrroles onto nitrones and N-hydroxylamines to afford selectively either dipyrromethanes or dipyrromethenes. This methodology has been the starting point of the first study in this manuscript. We have developed a pathway for the synthesis of Prodigiosin's analogues. These analogues, substituted in meso position, have never been studied. In the first stage, it was necessary to develop a synthetic sequence leading to the 4-methoxy-2, 2'-bipyrrole, a precursor of Prodigiosin. We also developed a sequence allowing access to unsymmetrical 2,2'-bipyrroles. These building blocks are used in the chemistry of expanded porphyrins. These pyrrole macrocycles show applications in many fields like conductive polymers, liquid crystals and other biological applications. Our methodology allows the formation of bipyrroles in six steps from readily available substrates with overall yields of up to 20%. This method has also been extended to the synthesis of terpyrrole.

Exposing Amyloid Beta 1-42 to neurons causes cell death. When carbonic anhydrase inhibitors (e.g. methazolamide or acetazolamide) are introduced along with 1-42 in a similar experiment, cell apoptosis is disrupted. However, when non-CA inhibitors are tested, (e.g. the indole derivative melatonin), the same disruption occurs. Are these carbonic anhydrase inhibitors acting on the same or a different pathway? One way to study the molecular mechanisms of these small molecule inhibitors is to modify their chemical structure. In this sense, when acetazolamide is methylated, apoptosis is resumed (Fossati et al., 2016). Finding a way to create N-methyl acetazolamide and N-methyl methazolamide through methylation procedures will lead to a better understanding of the pathways involved in neuronal apoptosis triggered by the Abeta peptide.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-01-14T18:45:29Z No. of bitstreams: 1 ElisangelaBarrosDantas_DISSERT.pdf: 2503846 bytes, checksum: 2247d4ef46af947a6abf96eacbada2b4 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-01-15T17:53:14Z (GMT) No. of bitstreams: 1 ElisangelaBarrosDantas_DISSERT.pdf: 2503846 bytes, checksum: 2247d4ef46af947a6abf96eacbada2b4 (MD5) / Made available in DSpace on 2016-01-15T17:53:14Z (GMT). No. of bitstreams: 1 ElisangelaBarrosDantas_DISSERT.pdf: 2503846 bytes, checksum: 2247d4ef46af947a6abf96eacbada2b4 (MD5) Previous issue date: 2012-06-04 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / As ligas de n?quel s?o largamente utilizadas na produ??o de diversos materiais, sobretudo, aqueles que necessitam de resist?ncia mec?nica associada ? resist?ncia ? corros?o, como por exemplo, os a?os inoxid?veis. Outro uso do n?quel ? a produ??o de ligas sinterizadas a partir de p? de n?quel met?lico. Uma alternativa promissora ? produ??o de componentes sinterizados de n?quel com importante redu??o de custos do material de partida ? o emprego de misturas de p?s do tipo NiNiO. Este trabalho teve por objetivo o estudo da redu??o in situ de NiO durante a sinteriza??o de misturas Ni/NiO produzidas por metalurgia do p?. As misturas foram processadas pela t?cnica de metalurgia do p? e pr?-sinterizadas em atmosfera redutora de hidrog?nio. As amostras Ni+15%NiO, Ni+25%NiO e Ni+35%NiO foram estudadas e comparadas com amostras sinterizadas apenas a partir de Ni met?lico. Foram realizados ensaios dilatom?tricos para estudo das condi??es de sinteriza??o das misturas. O material consolidado foi analisado quanto ? sua microestrutura e microdureza. As curvas de dilatometria mostraram que a adi??o de ?xido de n?quel em todas as composi??es ativa a sinteriza??o das misturas estudadas. Nos testes de microdureza foram realizadas indenta??es em pontos distintos da superf?cie das amostras. Todas as composi??es apresentaram valores de microdureza pr?ximos ao do material consolidado a partir de n?quel met?lico. Contudo, as amostras contendo Ni+35%NiO, apresentaram grande dispers?o desses valores. A an?lise microestrutural do material mostrou uma maior concentra??o de vazios e presen?a de ?xidos residuais nas misturas da composi??o Ni+35%NiO. As amostras contendo Ni+15%NiO apresentaram caracter?sticas microestruturais e propriedades mec?nicas semelhantes ao n?quel met?lico consolidado sob as mesmas condi??es das composi??es estudadas neste trabalho e, portanto, apresentaram grande potencial para produ??o de ligas sinterizadas de n?quel / The nickel alloys are widely used in the production of various materials, especially those that require mechanical strength characteristics associated with resistance to corrosion, for example, the stainless steel. Another use is the production of nickel alloy sintered from powder of metallic nickel. A promising alternative for the production of sintered components of nickel with an important reduction in costs of starting material is the use of mixtures of powders of Ni-NiO. This work aimed to study in situ reduction of NiO during sintering mixtures of Ni / NiO produced by powder metallurgy. The nickel mixtures have been processed by the technique of powder metallurgy and were pre-sintered in an oven under plasma reducing atmosphere of hydrogen. Mixtures Ni +15%NiO, Ni +25%NiO and Ni +35%NiO were studied and compared with samples consisting only of metallic Ni. Dilatometric tests were performed to study the sintering conditions of the mixtures. The consolidated material was analyzed for their microstructure and microhardness. Dilatometry graphs showed that the addition of nickel oxide in all compositions the active sintering the mixtures studied. In tests of microhardness indentations were made at different points of the sample surface. All compositions showed microhardness values close to the consolidated material from metallic nickel. However, sample containing Ni+35% NiO, showed a large dispersion of the values of microhardness tests performed at different points of the sample surface. Microstructural analysis of the material showed a higher concentration of voids and the presence of oxides in the waste composition of the mixtures Ni 35% NiO. The samples containing Ni+15%NiO showed microstructural characteristics and mechanical properties similar to metallic nickel consolidated under the same conditions of the compositions studied in this work and therefore had great potential for production of sintered nickel alloys

Made available in DSpace on 2014-12-17T14:53:00Z (GMT). No. of bitstreams: 1 CarlaSLT_TESE.PDF: 1695923 bytes, checksum: e946c937704d50c5dddffb25bf8ddd4c (MD5) Previous issue date: 2010-12-29 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Trabalhos anteriores propuseram modelos para controle on-line para vari?veis em produ??es finitas, que visam minimizar o custo m?dio esperado por item produzido. Por?m, em muitos casos a qualidade do produto n?o ? atestada atrav?s de mensura??o. Neste trabalho ser? proposta uma abordagem para monitoramento do n?mero de n?o-conformidades dos itens inspecionados para produ??es finitas, atrav?s de amostras n?o-unit?rias. A taxa m?dia de n?o-conformidades por itens ? considerada par?metro da distribui??o de Poisson. Ser?o utilizadas propriedades de uma cadeia de Markov finita de estados discretos, para determinar as probabilidades de mudan?a de estado, a serem consideradas nas express?es de custo, a m de determinar a estrat?gia ?tima de monitora??o, obtida atrav?s da otimiza??o dos par?metros: intervalo amostral (m) e o tamanho amostral (r). Os par?metros ser?o obtidos computacionalmente, atrav?s de busca direta, a fim de minimizar a express?o do custo m?dio por item produzido. O processo inicia (ou reinicia) no Estado I, onde a frequ?ncia m?dia de defeituosos ? 0; ap?s a mudan?a para o Estado II, a frequ?ncia m?dia de n?o-conformidades aumenta para 1. A mudan?a de estado, a cada item produzido, ocorre segundo uma distribui??o geom?trica. A cada ciclo de inspe??o, inspeciona-se o m-?simo item, mais (r-1) itens retrospectivamente. ? monitorado o n?mero de n?o-conformidades (D) de cada item e se, entre os r itens inspecionados, todos obedecerem ? regra de decis?o (D < DELTA), onde DELTA representa o limite superior de especifica??o, o processo continua operando; caso contr?rio, o processo ? parado para ajuste. Os itens inspecionados ser?o descartados depois da inspe??o, somente quando h? parada no processo para ajuste. Um lote adicional, que n?o sofrer? inspe??o, ser? produzido para completar a encomenda do cliente.

Trotz aller Fortschritte in Diagnostik und Behandlung bleibt die Therapie von BrustkrebspatientInnen – gerade bei tripelnegativen Mammakarzinomen, die nicht auf eine hormonelle Therapie ansprechen – eine Herausforderung. Eine wachsende Zahl von Belegen spricht dafür, dass Cadherine nicht nur in physiologischen Prozessen wie der Embryogenese, sondern auch in pathologischen Prozessen wie der Tumorprogression eine Rolle spielen, was diese Moleküle zu potenziellen Zielen einer targetspezifischen Tumortherapie macht. Im Mausmodell konnte bislang gezeigt werden, dass eine Herabregulierung von N-Cadherin das Mammakarzinomwachstum in vivo hemmt. N-Cadherin wird auch in humanem Brustkrebsgewebe aberrant exprimiert. In nur schwach invasiven Brustkrebszelllinien führte eine Überexpression von N-Cadherin in vitro zu einer Steigerung des migratorischen und invasiven Verhaltens der Zellen. In dieser Arbeit wurde an der humanen Mammakarzinomzelllinie SUM149PT der Einfluss einer Defizienz von N-Cadherin auf die Expression von E- und VE-Cadherin, sowie auf das Proliferations-, Migrations- und Invasionsverhalten der Zellen in vitro untersucht. Eine Charakterisierung der humanen Brustkrebszelllinien SUM149PT, der aus ihr hervorgegangenen mit shRNA transduzierten N-Cadherin-Knock-down-Klone und der Scramble-Kontrollen fand mittels Western Blot, RT-PCR, q-PCR und Immunfluoreszenzanalysen statt. Das Proliferationsverhalten der Zelllinien wurde mit Hilfe von Verdopplungszeitbestimmungen und Sphäroidversuchen analysiert. Um den Einfluss einer N-Cadherin-Defizienz auf die Migration zu untersuchen, wurden auf dem Prinzip der Boyden-Chamber basierende Versuche, sowie in vitro Wundheilungsversuche durchgeführt. Auch die Invasionsversuche basierten auf dem Prinzip der Boyden-Chamber. Eine zusätzliche Beschichtung mit Matrigel simulierte hier die extrazelluläre Matrix. Bei den Untersuchungen mittels Western Blot, RT-PCR und q-PCR wurde deutlich, dass die N-Cadherin defizienten Klone keine Veränderung in der E-Cadherin-Expression, jedoch interessanterweise eine starke Herabregulierung von VE-Cadherin aufweisen. Bei den Immunfluoreszenzanalysen wiesen SUM149PT und Scramble-Kontrollen eine Expression von N-Cadherin in der Zellmembran auf, während die N-Cadherin-defizienten Klone keine N-Cadherin-Expression zeigten. Wie schon in Western-Blot, RT-PCR und q-PCR zeigten die N-Cadherin defizienten Klone im Vergleich zu den SUM149PT und den Scramble-Kontrollen keine veränderte E-Cadherin-Expression. Bei Einsatz eines VE-Cadherin-Anitkörpers zeigten die N-Cadherin defizienten Klone keine Expression von VE-Cadherin, während SUM149PT und Scramble eine deutliche VE-Cadherin-Expression in der Zellmembran zeigten. Daraus lässt sich schließen, dass N-Cadherin möglicherweise die Expression von VE-Cadherin beeinflussen kann. In der Verdopplungszeitbestimmung ließen sich keine signifikanten Änderungen des Wachstumsverhaltens zwischen N-Cadherin defizienten Klonen und Kontrollzelllinien nachweisen. Um auch das dreidimensionale Wachstum der Zellen zu untersuchen, wurden Sphäroidversuche durchgeführt. Alle untersuchten Zelllinien bildeten stabile Sphäroide aus, die jedoch einen großen Saum an nicht integrierten Zellen aufwiesen. Es zeigte sich kein signifikanter Unterschied im Sphäroidwachstum zwischen N-Cadherin defizienten Klonen und Kontrollzelllinien. Die Herabregulierung von N-Cadherin scheint in humanen Mammakarzinomzelllinien in vitro keinen Einfluss auf das Wachstumsverhalten der Zellen zu haben. Im Boyden-Chamber-Assay zeigte sich, dass eine Herabregulierung von N-Cadherin bei der humanen Mammakarzinomzelllinie SUM149PT in vitro zu einer signifikanten Verringerung des Migrationspotenzials führt. Dieses Ergebnis wurde beim in vitro Wundheilungsversuch bestätigt. Hier zeigten die N-Cadherin defizienten Klone eine signifikant verringerte Flächenbedeckungsrate – also eine signifikant verringerte Geschwindigkeit mit der die aufeinander zuwanderndern Zellen eine freie Fläche bedecken – als die Kontrollzelllinien. Im Invasionsversuch zeigten die N-Cadherin defizienten Klone im Vergleich zu den Kontrollen eine hochsignifikant verringerte Invasivität. Eine Herabregulierung von N-Cadherin bei der humanen Mammakarzinomzelllinie SUM149PT führt also in vitro zu einer signifikanten Verringerung des Invasionspotenzials. Die Ergebnisse dieser Arbeit lassen den Schluss zu, dass N-Cadherin bei humanen Mammakarzinomzelllinien durch seinen positiven Einfluss auf Migration und Invasion der Zellen eine große Rolle beim Prozess des invasiven Wachstums und der Metastasierung spielt. Eine Anti-N-Cadherin-Therapie könnte auch bei BrustkrebspatientInnen, gerade mit tripelnegativen Mammakarzinomen, neue Möglichkeiten der Behandlung eröffnen. / Breast cancer is the most common cancer type for women and the most frequent type after lung cancer overall. In spite of all improvements in clinical diagnostics and treatment, the therapy of patients with breast cancer remains a difficult task. Therefore it is important to find new strategies of treatment, especially for patients with triple negative breast cancer that does not respond to hormone therapy. There is growing evidence that cadherins are not only important in physiological processes like embryogenesis but also in pathological processes like tumour progression. A so called cadherin switch has been implicated in carcinogenesis, in particular the loss of E-cadherin and the upregulation of N-cadherin. Thus, these molecules are an interesting subject of studies and a potential target for specific cancer therapy. It was shown previously that efficient silencing of N-cadherin in murine breast cancer cells inhibits tumour growth in vivo. N-cadherin is also aberrantly expressed in human breast cancer tissue. An overexpression of N-Cadherin in breast cancer cells enhances tumour cell motility, migration and invasion in vitro. In this thesis the impact of N-cadherin deficiency on the expression level of E- and VE-cadherin as well as its effect on proliferation, migration and invasion behaviour of breast cancer cells in vitro was investigated. The human breast cancer cell line SUM149PT, its N-cadherin deficient clones and scramble controls were characterised by Western Blot, RT-PCR, q-PCR and immunofluorescence staining. To analyse the impact of N-cadherin on the proliferation behaviour doubling time proliferation assays and spheroid assays were performed. A Boyden chamber assay and an in vitro wound healing assay were performed to investigate the effect of N-cadherin deficiency on cell migration. Furthermore, a Boyden chamber assay with a coating of Matrigel was used to study the invasion potential of the N-cadherin deficient cell clones and control cell lines. The results of Western Blot, RT-PCR and q-PCR show that silencing of N-cadherin expression in SUM149PT cells had no influence on E-cadherin expression, but significantly decreased VE-cadherin expression. With immunofluscence staining it was evidenced that SUM149PT and scramble controls express N-cadherin in the cell membrane, whereas N-cadherin deficient clones do not show any N-cadherin expression. There was no difference seen in E-cadherin expression between N-cadherin deficient clones and control cell lines. While SUM149PT and scramble controls expressed VE-cadherin in the cell membrane, N-cadherin deficient clones did not express VE-cadherin. These results suggest that N-cadherin may have an influence on the expression of VE-cadherin. No significant alteration in proliferation behaviour between N-cadherin deficient clones and controls could be shown with the doubling time proliferation assay. To investigate the impact of N-cadherin on the three-dimensional growth spheroid assays were performed. All cell lines formed stable speroids, however fringes of many not incorporated cells were found. There was no difference in spheroidgrowth between N-cadherin deficient clones and control cell lines. No influence of N-cadherin on cell proliferation in vitro could be seen. With the help of Boyden chamber assays it was shown that silencing of N-cadherin in the human breast cancer cell line SUM149PT results in a significant decrease of migration potential in vitro. This was confirmed with an in vitro wound healing assay. N-cadherin deficient clones showed a significantly reduced surface coverage rate than SUM149PT and scramble controls. A significant decrease of invasive cells in N-cadherin deficient clones in comparison to the controls was revealed in invasion assays. Thus silencing of N-Cadherin in the human breast cancer cell line SUM149PT leads to a significantly decreased invasion potential in vitro. The results presented this thesis provide evidence that N-cadherin is involved in invasive tumour progression and metastasis of the human breast cancer cell line SUM149PT by promoting cell migration and invasion. Anti-N-cadherin strategies could thus be a potential target specific therapy of cancer patients, particularly with triple negative breast cancer.

Foi efetuado o estudo cristalográfico do ácido N-acetilantranílico (C9H903N) sendo apresentados os fundamentos teóricos e computacionais para determinar a sua estrutura. Os dados de 899, reflexões independentes, foram obtidos num difratômetro automático CADA KAPPA com radiação ka do cobre e processados pelo conjunto de programas denominado \"STRUCTURE DETERMINATION PACKAGE\", (SDP), desde sua redução ate à conclusão o trabalho; Os cristais do ácido N-acetilantranílico são incolores e em forma de placas retangulares e pertencem ao grupo espacial Fdd2 com a = 10,845 (9) R , b = 30,204 (7) R , c = 10,575 (4) R , a = 3 = y = 90° , V = 3464 (5) R3, d cal = 1,374 a/cm3, d obs 1,36 g/cm 3 , Z = 16 moléculas por cela. As moléculas do ácido N-acetilantranílico são planares e formadas Por um anel benzênico ao qual estão ligados em carbonos adjacentes um grupo carboxila e um radical acetamida. Uma ponte de hidrogênio entre a carboxila e a acetamida fazem a ligação entre as moléculas. As moléculas assim ligadas formam uma cadeia que se desenvolve em planos alternados, aproximadamente (301) e (J01). Cadeias vizinhas desenvolvem-se nos planos (501) e (301) / In this thesis it is intended to study the N-acetyl Anthranilic Acid (C9 H9 O 3 N). Fundamentals of theoretical and computational are presented in order to determine its Structure. Data of 899 independent reflexions were obtained by -using the Automatic Difratometer (CADA KAPPA) with a ka copper radiation. \"STRUCTURE DETERMINATION PACKAGE\" (SDP) Programs were used during - this work. N-Acetyl Anthranilic Acid Crystals are in the form of rectangular plates which are transparent and belong to the spatial groupFdd2 where a = 10.845 (9) R , b = 30.204 (7) R , c 10.575 (4) R , a = 8 = y = 90 0 , V = 3464 (5) g3 , d cal = 1,374 \' g/cm 3 , d obs = 1,36 g/cm3 , Z = 16 molecules by cell. The N-Acetyl Anthranilic Acide Molecules is formed \' by a benzenic ring which is bonded to adjacent carbon, a carboxyl and a acetamide groups. Two molecules are bonded through Hydrogen bridge between the carboxyl and acetamide. The molecules are bonded such a way to form a chain with alternate planes (301) and (301). The neibouring chains are in connection with the first chain formed with the planes (301) \' and (301)