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In dieser Arbeit wurde ein Verfahren zur rückführbaren Proteinquantifizierung unter Verwendung von Schwefelisotopenverdünnung mit induktiv gekoppelter Plasmamassenspektrometrie (ID-ICP-MS) entwickelt. Die Methode dient zur zuverlässigen Quantifizierung laborinterner Proteinstandards. Sie eignet sich nur zur Analyse reiner Proteine, deren Stöchiometrie bekannt ist, da die Proteinkonzentration aus dem Schwefelgehalt bestimmt wird. Nicht proteingebundener Schwefel wird durch Membranfiltration von der Proteinfraktion abgetrennt und mit ID-ICP-MS quantifiziert. Der Gesamtschwefelgehalt in der Probe wird ebenfalls mittels ID-ICP-MS quantifiziert und um die Menge an ungebundenem Schwefel korrigiert. Die Optimierung der Probenvorbereitung zeigte, dass ein Aufschluss die Unsicherheit des Ergebnisses verbessert. Für die Methodenentwicklung wurden ein zertifiziertes Rinderserumalbumin-Referenzmaterial (BSA), sowie ein kommerzielles Avidin verwendet. Die Bestimmung der Proteinmassenfraktionen und Unsicherheitsbudgets erfolgte nach Korrektur für den ungebundenen Schwefel (0,4 % für BSA, 30 % für Avidin). Die Methode wurde auf das Tau-Protein angewendet, das ein Biomarker für die sogenannten „Tauopathien“ ist – eine Gruppe neurodegenerativer Krankheiten, die die Alzheimer-Krankheit und die frontotemporale Demenz einschließen. Durch Verwendung eines isotopenangereicherten Spikes, der an das SI-System angebunden ist, wurde die metrologische Rückführbarkeit für den Massenanteil des Tau-Proteins erreicht. Dieser Tau-Standard wurde zur absoluten Quantifizierung toxischer transgener Tau-Spezies in der löslichen Hirnfraktion transgener Mäuse verwendet. Die entwickelte Methode ist für die rückführbare Quantifizierung von Proteinen zur Verwendung als Standards in der biologischen und medizinischen Forschung anwendbar. Sie zeigte eine ähnliche Präzision wie etablierte Verfahren, erforderte jedoch weniger Probenvorbereitung und keine spezies-spezifischen Standards. / In this work, a method for traceable protein quantification using sulphur isotope dilution inductively coupled plasma mass spectrometry (ID-ICP-MS) was developed. The method is intended for the reliable quantification of in-house protein standards. It is only suited for pure protein formulations for proteins of known stoichiometry because the protein concentration is determined from the sulphur content. Non-protein bound sulphur is separated from the protein fraction by membrane filtration and is quantified by ID-ICP-MS. The total sulphur content in the sample is as well quantified by ID-ICP-MS and corrected for the amount of non-protein bound sulphur. Optimisation of the sample preparation showed that digestion improves the uncertainty of the result. For method development, a certified reference material bovine serum albumin (BSA) and a commercial avidin were used. The protein mass fractions with full uncertainty budgets were determined after correction for unbound sulphur. The developed procedure was applied to the tau protein, which is a biomarker fort he so-called „tauopathies“ – a group of neurodegenerative diseases including Alzheimer’s disease and Frontotemporal dementia. By employing an isotopically enriched spike, which is linked to the SI system, full metrological traceability was achieved for the tau mass fraction. The mass fraction of (0.328 ± 0.036) g kg-1 for tau was determined by ID-ICP-MS and confirmed by amino acid analysis. This tau standard was used for the absolute quantification of toxic transgenic tau species in the soluble brain fraction of transgenic mice. The developed method is applicable for the traceable quantification of proteins for use as standards in biological and medical research. The precision of the method was similar to established absolute protein quantification procedures while requiring less sample preparation and no species-specific standards.

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Made available in DSpace on 2015-04-14T13:54:59Z (GMT). No. of bitstreams: 1 414799.pdf: 393719 bytes, checksum: e09801b5cd51426472def038700b8bf1 (MD5) Previous issue date: 2009-05-15 / O objetivo do presente trabalho ? estabelecer a rela??o entre sensa??o e percep??o na teoria do conhecimento emp?rico de Thomas Reid, efetuando uma an?lise do realismo direto em epistemologia. O tema ? abordado com base, principalmente, em duas de suas grandes obras, An Inquiry into the Human Mind on the Principles of Common Sense e Essays on the Intellectual Powers of Man, mas sem deixar de apresentar as ideias de grandes fil?sofos contempor?neos, int?rpretes de Reid como John Greco, James van Cleve, entre outros. No primeiro momento, ressalta-se a cr?tica de Reid ? teoria das ideias, estabelecendo-se em seguida uma distin??o preliminar entre sensa??o e percep??o. A teoria da sensa??o e a teoria da percep??o s?o ent?o tratadas separadamente, conforme a metodologia adotada por Reid. Para finalizar, o ?ltimo cap?tulo trata do realismo direto de Thomas Reid, que, na teoria da percep??o, demonstra a capacidade que a mente possui de se conectar diretamente com o mundo, sem precisar de qualquer outro elemento, como a pr?pria "ideia".

Submitted by Filipe dos Santos (fsantos@pucsp.br) on 2017-10-03T12:32:00Z No. of bitstreams: 1 Luna Neide Macedo Gutierres.pdf: 2504303 bytes, checksum: 02a4e9360ce4e69a8c820a68f718d39a (MD5) / Made available in DSpace on 2017-10-03T12:32:00Z (GMT). No. of bitstreams: 1 Luna Neide Macedo Gutierres.pdf: 2504303 bytes, checksum: 02a4e9360ce4e69a8c820a68f718d39a (MD5) Previous issue date: 2017-09-19 / The world faces exponential data growth. Data is created by smart devices, RFID technologies (Radio-Frequency IDentification), sensors, social networks, video surveillance and more. These generated data are no longer considered static, whose usefulness ends after the purpose of the collection is reached, they have become the raw material of the business, a vital economic resource, used to create a new form of economic value. Then comes the concept of “big data”. The objective of this research is to raise the discussion about the concept of big data, drawing from the current literature definitions that offer subsidies for the understanding of its real meaning and impact in the generation of useful ideas and goods and services of significant value. However, because it is a recent theme, the available literature is scarce. It is an applied purpose research with a descriptive purpose and uses the qualitative method of approach. It has by type of research the review of the literature for the theoretical basis, and also the study review of two cases through an exploratory research to collect the data to be analyzed. It seeks to confront the theory with the identified hypotheses and practices, to assess its adherence, arriving at informed conclusions, and to suggest future studies that may continue this line / O mundo enfrenta um crescimento exponencial de dados. Dados são criados por dispositivos inteligentes, tecnologias RFID (Radio-Frequency IDentification), sensores, redes sociais, vigilância por vídeo e muito mais. Esses dados gerados não são mais considerados estáticos, cuja utilidade termina depois que o objetivo da coleta é alcançado, eles se tornaram a matéria-prima dos negócios, um recurso econômico vital, usado para criar uma nova forma de valor econômico. Surge então o conceito de “big data”. O objetivo desta pesquisa é levantar a discussão sobre o conceito de big data, extraindo da literatura atual definições que ofereçam subsídios para o entendimento de seu real significado e impacto na geração de ideias úteis e bens e serviços de valor significativo. Entretanto, por ser um tema recente, a literatura disponível é escassa. É uma investigação de finalidade aplicada, com um objetivo descritivo e utiliza o método qualitativo de abordagem. Tem por tipo de pesquisa a revisão da literatura para a fundamentação teórica, e também a revisão de estudo de dois casos através de pesquisa exploratória para a coleta dos dados a serem analisados. Busca confrontar a teoria com as hipóteses e práticas identificadas, para avaliar sua aderência, chegando em conclusões fundamentadas, além de sugerir estudos futuros que podem dar continuidade a esta linha abordada

A leptospirose é uma zoonose de importância mundial causada por bactérias do gênero Leptospira. No Brasil, a maioria dos casos é causada por L. interrogans sorovar Copenhageni. O objetivo destre trabalho foi identificar adesinas de leptospira pela técnica de Phage display. Bibliotecas com fragmentos genômicos resultaram na idendificação de ligantes de leptospira com afinidade por tecidos de hamster. Uma varredura dessas bibliotecas contra heparan sulfato proteoglicano (HSPG) identificou como ligantes as proteínas LigA e LigB. Proteínas recombinantes foram produzidas e submetidas à ligação às células de mamíferos e aos componentes de matriz extracelular. LigB recombinante foi capaz de se ligar ao HSPG, à heparina e às células de mamíferos. HSPG e heparina foram capazes de reduzir significativamente a interação dessa proteína com as células. Estes resultados evidenciam o papel de proteínas da leptospira na sua interação com o hospedeiro e ilustram a possibilidade do uso da técnica de phage display para identificar possíveis adesinas. / Leptospirosis is a worldwide important zoonosis caused by bacteria of the genus Leptospira. In Brazil, most cases is caused by L. interrogans serovar Copenhageni. Our goal was to identify leptospiras adhesins by phage display technique. Libraries of genomic fragments resulted in the identification of ligands with affinity for leptospiras hamster tissues. Screening these libraries against heparan sulfate proteoglycan (HSPG) identified the proteins LigA and LigB. Recombinant proteins were produced and subjected to binding to mammalian cells and extracellular matrix components. LigB recombinant was able to bind to HSPG, heparin and mammalian cells. HSPG and heparin were able to significantly reduce the interaction of this protein with cells. These results highlight the role of leptospiras proteins in its interaction with the host and illustrate the possibility of the use of phage display technique to identify potential adhesins.

Submitted by Filipe dos Santos (fsantos@pucsp.br) on 2017-12-07T11:14:05Z No. of bitstreams: 1 Nelson Maria Brechó da Silva.pdf: 1743917 bytes, checksum: 4264c2b3b0b81216f0b0d958b0f697f6 (MD5) / Made available in DSpace on 2017-12-07T11:14:05Z (GMT). No. of bitstreams: 1 Nelson Maria Brechó da Silva.pdf: 1743917 bytes, checksum: 4264c2b3b0b81216f0b0d958b0f697f6 (MD5) Previous issue date: 2017-11-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / This thesis has a target to show the friendship as exercise of judgment in Montaigne, inspired by his friend La Boetie. From this famous friendship, which was disrupted by the death of La Boetie, follows, on the one hand, a study of the concept of friendship on Montaigne’s Essais and his main partners. On the other hand, the humanistic traits of the essayist in the passage from alterity to identity. The interpretation of the text allows reflections about friendship according to the image of the writing, painting, transfiguration and mirror / A presente tese pretende situar a amizade como um exercício do julgamento em Montaigne, que evoca a figura de seu amigo La Boétie. A partir dessa célebre amizade que foi rompida com a morte de La Boétie, segue-se, por um lado, uma análise do conceito de amizade nos Essais de Montaigne e de seus interlocutores. Por outro, os traços humanísticos do ensaísta na passagem da alteridade à identidade. A interpretação do texto permitirá reflexões sobre o sentido maior da amizade à luz da imagem da escrita, da pintura, da tranfiguração e do espelho

Univ., Diss.-2006--Bochum, 2005. / Literaturverz. S. [154] - 171.

Univ., Diss.--Utrecht, 2006.

Univ., Habil.-Schr.--Rostock, 2004.