Made available in DSpace on 2014-12-17T14:13:22Z (GMT). No. of bitstreams: 1 SebastiaoDSF.pdf: 475002 bytes, checksum: d28adb2821adc67501d7e19f42e9b6a3 (MD5) Previous issue date: 2007-08-16 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Several clinic evaluations have been possible with radiobiocomplexes labeled with technetium-99m (99mTc). Some natural and synthetic drugs are capable of to interfere on the labeling of blood constituents with 99mTc, as well as on the biodistribution of radiobiocomplexes. Authors have also reported about the toxicity of several natural products. The aim of this study was to compare the effects of the Mentha crispa (hortel?) and of the Hypericum perforatum (hip?rico) in different experimental models. On the labeling of red blood cells (RBC) and plasma and cellular proteins with 99mTc, both extracts were capable of to decrease the radioactivity percentage on the cellular compartment and on the fixation on plasma and cellular proteins. On the morphometry of the RBC, only the hortel? was capable to alter the shape and the perimeter/area ratio of the RBC. On the biodistribution of the radiobiocomplex sodium pertechnetate (Na99mTcO4), the hortel? increased the Na99mTcO4 distribution in the kidney, spleen, liver and thyroid, meanwhile the hip?rico decreased the Na99mTcO4 distribution in the bone, stomach, lungs and thyroid, and increased the Na99mTcO4 distribution in the pancreas. On the bacterial cultures survival, the hip?rico was capable of to protect the bacteria against the stannous chloride (SnCl2) effect. The hip?rico did not alter the topology of plasmidial DNA and did not protect the plasmidial DNA against the SnCl2 action. Probably, the effects presented by both extracts could be due to chemical compounds of the extracts that could alter the morphology of the RBC and the plasma membrane ions transport, and/or by phytocomplexes that could be formed with different effects dependent on the biological system considered / Avalia??es cl?nicas t?m sido poss?veis com radiobiocomplexos marcados com tecn?cio-99mTc (99mTc). Drogas naturais ou sint?ticas s?o capazes de interferir na marca??o de estruturas sangu?neas com 99mTc, assim como na biodistribui??o de radiobiocomplexos. Tamb?m tem sido descrita a toxicidade de v?rios produtos naturais. O objetivo deste estudo foi comparar o efeito dos extratos de Mentha crispa (hortel?) e de Hypericum perfloratum (hip?rico) em diferentes modelos experimentais. Na marca??o de estruturas sang??neas com 99mTc verificou-se que ambos os extratos foram capazes de diminuir a radioatividade no compartimento celular, nas prote?nas plasm?ticas e celulares. Na morfometria das hem?cias, apenas a hortel? foi capaz de alterar a forma e a rela??o per?metro/?reas das hem?cias. Na biodistribui??o do radiobiocomplexo pertecnetato de s?dio (Na99mTcO4) a hortel? aumentou a capta??o do Na99mTcO4 no rim, no ba?o, no f?gado e na tire?ide, enquanto que o hip?rico diminuiu a capta??o do Na99mTcO4 no osso, no est?mago, no pulm?o e na tire?ide, e aumentou no p?ncreas. Na sobreviv?ncia de culturas bacterianas o hip?rico foi capaz de proteger a bact?ria do efeito danoso do cloreto estanoso (SnCl2). O hip?rico n?o alterou a topologia nem protegeu o DNA plasmidial da a??o do SnCl2. Provavelmente os efeitos apresentados por ambos os extratos poderiam ser explicados por subst?ncias presentes nos extratos que poderiam alterar a morfologia das hem?cias, o transporte de ?ons pela membrana e/ou formar fitocomplexos. O estudo teve car?ter multidisciplinar com a participa??o das seguintes ?reas do conhecimento: Radiobiologia, Bot?nica, Endocrinologia, Fitoterapia e Hematologia

O fator VIII de coagulação (FVIII), recombinante ou purificado de plasma, é o biofármaco necessário para o tratamento da hemofília A, a doença hemorrágica mais freqüente em humanos. O método tradicional para a purificação de FVIII parte de crioprecipitado de plasma e precipitação alcoólica. No Instituto Butantan, foi proposto um método alternativo, utilizando somente cromatografia para esta purificação. Este projeto teve por objetivo comparar dois métodos cromatográficos de purificação do FVIII: 1 - gel filtração direta do plasma e 2 - pré-purificação de FVIII do plasma por cromatografia de troca aniônica, seguida de gel filtração. A purificação foi analisada por dosagens de atividade específica de FVIII e presença de outras proteínas da cascata de coagulação nas frações de cromatografia. Foram realizadas clonagem de fragmentos gênicos de FVIII e expressão de fragmentos protéicos para imunização de animais. Os soros com anticorpos policlonais anti-FVIII foram usados em ensaios de \"western blot\" para detectar as cadeias de FVIII ou degradação. / Coagulation factor VIII (FVIII), recombinant or purified from plasma, is the biopharmaceutical used for treatment of haemophilia A, the most frequent human hemorrhagic disorder. The traditional method used for purification of FVIII starts from plasma cryoprecipitate and alcoholic precipitation. The Instituto Butantan proposed an alternative methodology using only chromatography for FVIII purification. The main objective of this project was to compare two chromatographic methods for FVIII purification: 1 - direct plasma gel filtration and 2 - pre-purification of FVIII by anion exchange chromatography, followed by gel filtration. The purification process was analyzed by determination of FVIII specific activity and detection of other coagulation factors co eluting in chromatographic fractions. Fragments of FVIII gene were cloned and protein fragments were expressed for animal immunization. Sera with polyclonal antibodies anti-FVIII were used in western blots assays to detect FVIII chains or its degradation.

La sang és un subproducte amb un alt potencial de valorització que s'obté en quantitats importants en els escorxadors industrials. Actualment, la majoria de sistemes de recollida de la sang no segueixen unes mesures d'higiene estrictes, pel que esdevé un producte de baixa qualitat microbiològica. Conseqüentment, l'aprofitament de la sang és una sortida poc estimulant des del punt de vista econòmic, ja que acostuma a perdre les qualitats que permetrien l'obtenció de productes d'alt valor afegit. El capítol I del present treball s'inclou dins d'un projecte que proposa la inoculació de bacteris de l'àcid làctic (LAB) com un cultiu bioconservador de la sang, un sistema senzill i de baix cost que cerca l'estabilitat de la sang, tant microbiològica com fisicoquímica, durant el període del seu emmagatzematge. El capítol II s'emmarca dins d'un projecte que cerca la millora de l'aprofitament integral de la sang que, en el cas de la fracció plasmàtica, es centra en l'estudi de la funcionalitat dels seus principals constituents. Conèixer la contribució dels components majoritaris ha de permetre la millora de la funcionalitat dels ingredients alimentaris derivats. Els resultats presentats en aquesta tesi poden ajudar a la valorització de la sang porcina d'escorxadors industrials, mitjançant els coneixements adquirits pel que fa a la millora del seu sistema de recollida i del desenvolupament d'ingredients alimentaris amb interessants propietats funcionals. / Blood is a by-product obtained in large amounts in industrial slaughterhouses with a high potential of valorisation. Currently, most of the blood collecting systems are not subjected to strict hygienic measures hence it becomes a product with low microbiological quality. As a result, the use of blood for consumption purposes is not a stimulating prospect from an economic point of view, because the intrinsic worth allowing the development of high value-added products is normally lost.Chapter I of the present dissertation is included within a research project that suggests lactic acid bacteria (LAB) as a blood biopreservative culture: a simple, inexpensive system to keep the stability of blood both in terms of microbiological and physicochemical quality, during its storage.Chapter II is framed within a research project that investigates ways to improve the use of blood as a food ingredient, which, in the case of plasma, is focused on the functionality of its main protein constituents. The knowledge of the contribution of each constituent can be used to improve the functional properties of plasma-based ingredients. The results presented in this thesis dissertation may help the valorisation of porcine blood from industrial slaughterhouses, thanks to the acquired knowledge about the improvement of blood preservation and the development of plasma-based food ingredients with interesting functional properties.