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Pontos quânticos como sondas fluorescentes no estudo da carcinogênese em Células GliaisSEABRA, Maria Aparecida Barreto Lopes 24 March 2014 (has links)
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Previous issue date: 2014-03-24 / Glioblastoma (grau IV) é o tumor mais agressivo e infiltrante do sistema nervoso central (SNC),
que mostra uma série de mutações, bem como alto grau de vascularidade, polimorfismo e atipia
celular nuclear. Infelizmente, diagnóstico precoce de tumores cerebrais é difícil, uma vez que
ferramentas de imagem não são eficientes para o diagnóstico correto desses tipos de tumores,
levando a falhas no tratamento. Aqui nós descrevemos a síntese, caracterização e conjugação de
pontos quânticos ou quantum dots (QDs) de telureto de cádmio (CdTe) recobertos com tiol e
com o anticorpo contra a proteína ácida fibrilar glial (anti-GFAP), bem como, a preparação de
lipossomas. O método de congelamento e descongelamento foi utilizado para encapsular os QDs
em diferentes tipos de lipossomas e liberá-los em células tronco. Os lipossomas vazios e
contendo CdTe QDs foram caracterizados por microscopia de fluorescência, microscopia
eletrônica de transmissão, tamanho e potencial zeta. Os QDs CdTe-anti-GFAP foram utilizados
para um novo direcionamento in vivo e método de imagem para detecção do tipo de tumor
glioblastoma U87 xenotransplantado em cérebro de camundongos suíços machos. CdTe QDs não
conjugados e CdTe QDs conjugados foram utilizados para marcar U87 linha de células de tumor
in vitro e astrócitos saudáveis. A citotoxicidade dos CdTe QDs com fluorescência no verde (530
nm) e no vermelho (644 nm), foi avaliada utilizando MTT nas células U87. O crescimento do
tumor foi visualizado no interior do cérebro pela marcação com hematoxilina e eosina e mostrou
a entrega com sucesso nas células U87 no parênquima cerebral. CdTe QDs conjugados com anti
- GFAP foram injectados na região do tumor e a sua marcação na linhagem celular U87 foi
visualizada por microscopia de fluorescência, mostrando dupla marcação com especificidade
para vimentina em glioblastomas imunorreactivos. Em comparação com as células tumorais
U87, que facilmente foram marcadas com o CdTe QDs conjugados com anti- GFAP, observouse
que os astrócitos saudáveis mantidos em culturas primárias tiveram uma maior resistência à
sua marcação e foram fracamente marcados. Os resultados descritos aqui direcionam para novas
perspectivas na utilização de CdTe QDs na detecção de gliobastoma , sugerindo uma potencial
aplicação em cirurgia guiada por imagem. / Glioblastoma (grade IV) is the most aggressive and infiltrating tumor of the central nervous
system (CNS), showing a variety of mutations as well as high degree of vascularity, cell
polymorphism and nuclear atypia. Unfortunately, early diagnostic of brain tumors is hard, as
imaging tools are not efficient for proper diagnosis of these types of tumors, leading to treatment
failures. Here we describe the synthesis, characterization and conjugation of thiol-capped CdTe
with anti-glial fibrillar acidic protein (anti-GFAP) and preparation of liposomes. The freeze and
thaw method was used to encapsulate the QDs in different types of liposomes and deliver them
into stem cells. The empty and containing CdTe QDs liposomes were characterized by
fluorescence microscopy, transmission electron microscopy, size and zeta potential. A new in
vivo targeting and imaging method for U87 glioblastoma tumor type xenotransplanted into male
swiss mice brain using aqueous colloidal CdTe quantum dots conjugated to anti-GFAP (CdTeanti-
GFAP QDs) was developed. The red emitting CdTe QDs and the conjugated red-emitting
CdTe QDs were used to label U87 tumor cell line in vitro and health astrocytes. Toxicity of
isolated green (530 nm) and red (644 nm) emitting CdTe QDs, was evaluated using MTT assay
applied to U87 cells. The tumor growth was visualized inside the brain by the hematoxylin and
eosin staining and showed the successful delivery of the U87 cells into the brain parenchyma.
CdTe-anti-GFAP QDs were injected into the tumor region and their uptake by the U87 cell line
was visualized by fluorescence microscopy, showing specific double-labeling of vimentinimmunoreactive
glioblastoma. Compared to U87 tumor cells, which easily take up anti-GFAP
conjugated red-emitting CdTe QDs, healthy astrocytes kept in primary cultures offered more
resistance to their incorporation and were weakly labeled. The results reported here provide new
perspectives for using CdTe QDs in gliobastoma detection, suggesting their potential application
in imaging-guided surgery.
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Avaliação da atividade esquistossomicida do 3 - (4-clorobenzil)-5-(4- nitrobenzilideno)-imidazolidina-2,4-diona (FZ-4) e do Praziquantel frente a Schistosoma mansoni (cepa BH)Guabiraba e Silva, Polliana January 2006 (has links)
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Previous issue date: 2006 / O insucesso surgido na terapia da esquistossomose com o praziquantel, vem
acenando para uma necessidade de se buscar novas drogas para o tratamento desta doença
bem como a de se monitorar sua atividade frente cepas de Schistosoma. O objetivo deste
trabalho foi de avaliar a atividade esquistossomicida in vivo do Praziquantel (PZQ) e do
derivado imidazolidínico 3-(4-clorobenzil)-5-(4-nitrobenzilideno)-imidazolidina-2,4-diona
(FZ-4). Foram utilizados nove grupos de camundongos Swiss, com seis animais cada, que
foram infectados experimentalmente com 80 cercárias de S. mansoni (cepa BH), e tratados
por via oral durante cinco dias consecutivos com PZQ nas doses de 25, 50, 100 e 200
mg/Kg e com FZ-4 nas doses de 25, 50 e 100 mg/Kg, 50 dias após a infecção. Um grupo
controle foi utilizado para o PZQ e outro para o FZ-4. O praziquantel mostrou-se eficaz em
reduzir a carga parasitária dos animais infectados, com redução de 100 % nas doses de 100
e 200mg/Kg. A ED50 calculada foi de 14,75mg/Kg demonstrando que a cepa BH de S.
mansoni utilizada foi sensível ao Praziquantel. O FZ-4 nas doses utilizadas não mostrou ser
capaz de reduzir o número de vermes nem alterar o oograma nos animais infectados,
todavia na dose de 25mg/Kg apresentou significativo aumento no número de ovos
inviáveis com p < 0,05. Também foi capaz de reduzir o número de ovos por grama de fezes
quando analisados no sexto e décimo quarto dias após o término do tratamento. A análise
histopatológica indicou que o FZ-4 promoveu redução das áreas de fibrose e proteção da
mucosa intestinal da reação granulomatosa, ativando nódulos linfáticos locais. na dose de
100mg/Kg. Em conclusão a atividade esquistossomicida do praziquantel foi evidenciada,
mas não foi com o FZ-4, contudo este tenha reduzido o número de ovos do parasito nas
fezes
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Novas moléculas imidazolidínicas: potenciais candidatas a fármacos esquistossomicidasCristina Apolinário da Silva, Andréa 31 January 2008 (has links)
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Previous issue date: 2008 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A esquistossomose é uma doença parasitária crônica das regiões tropicais e
subtropicais e está associada a uma alta morbidade da população infectada. A
terapia da esquistossomose é feita apenas com um único fármaco, o
praziquantel, ativo contra todas as espécies de Schistosoma, e o oxamniquina
ativo apenas contra o Schistosoma mansoni. Contudo há relatos na literatura
de resistência do parasito ao tratamento para os dois fármacos, gerando uma
preocupação a nível mundial e levando a Organização Mundial de Saúde
(OMS) convocar a indústria e os pesquisadores a buscar novos fármacos com
esta finalidade. Neste contexto a busca de novos fármacos com atividade
esquistossomicida torna-se o objetivo deste trabalho, onde os derivados
imidazolidínicos das séries 5-benzilideno-3-benzil-4-tioxo-imidazolidin-2-ona
(LPSF/PT), 5-benzilideno-3-(4-metil-benzil)-imidazolidina-2,4-diona (LPSF/MS)
e 5-arilazo-4-tioxo-imidazolidin-2-ona (LPSF/PT) foram avaliados in vitro frente
a vermes adultos de Schistosoma mansoni (Cepa BH). Dos derivados
avaliados in vitro os que apresentaram atividade esquistossomicida, foram os
derivados da série 3-benzil-5-(arilazo)-4-tioxo-imidazolidin-2-ona (LPSF/PT).
Para a avaliação in vivo, o derivado 3-benzil-5-(4-cloro-arilazo)-4-tioxoimidazolidin-
2-ona (LPSF/PT05) foi administrado oralmente em Tween 80,
Tween/óleo/água e em dispersão sólida com 90 % PEG (Polietilenoglicol).
Apenas esta última apresentou efeito esquistossomicida. Foram administradas
quatro doses: 3, 10, 30 e 100 mg/kg. A redução do número de vermes
recuperados foi significativa para a dose de 10 e 30 mg/kg, aproximadamente
50 % e para a dose de 100 mg/kg a redução foi de 68%. A redução do número
de vermes na dose de 100 mg/kg corroborou com a resposta imune celular do
derivado e levantou a hipótese do seu efeito imunossupressor ser ao nível de
resposta Th1 e Th2. Somado a esses dados a resposta histopatológica revelou
uma modulação na resposta granulomatosa, produziu uma ação contra o dano
hepático causado pela infecção pelo S. mansoni
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Interação de Paracoccidioides com o hospedeiro: análises proteômicas de lavado broncoalveolar / Paracoccidioides interaction with the host: proteomic analysis of bronchoalveolar lavage fluidPigosso, Laurine Lacerda 09 May 2016 (has links)
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Previous issue date: 2016-05-09 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Paracoccidoides brasiliensis and Paracoccidioides lutzii, the etiologic agents of paracoccidioidomycosis, cause disease in healthy and immunocompromised persons in Latin America. We developed a method for harvesting P. brasiliensis yeast cells from infected murine lung to facilitate in vivo transcriptional and proteomic profiling. P. brasiliensis harvested at 6 h post-infection were analyzed using RNAseq and LC-MSE. In vivo yeast cells had 594 differentially expressed transcripts and 350 differentially expressed proteins. Integration of transcriptional and proteomic data indicated that early in infection (6 h), P. brasiliensis yeast cells underwent a shift in metabolism from glycolysis to β-oxidation, upregulated detoxifying enzymes to defend against oxidative stress, and repressed cell wall biosynthesis. Bioinformatics and functional analyses also demonstrated that a serine proteinase was upregulated and secreted in vivo. To our knowledge this is the first study depicting transcriptional and proteomic data of P. brasiliensis yeast cells upon 6 h post-infection of mouse lung. / Paracoccidoides brasiliensis e Paracoccidioides lutzii, os agentes etiológicos da paracoccidioidomicose, causam doenças em pessoas saudáveis e imunocomprometidas na América Latina. Desenvolvemos um método para coletar células de levedura de P. brasiliensis de pulmão murino infectado para facilitar o estudo do perfil transcricional e proteômico in vivo. P. brasiliensis recuperados após 6 horas de infecção foram analisados usando RNAseq e LC-MSE. As células de levedura após passagem in vivo apresentaram 594 transcritos expressos diferencialmente e 350 proteínas diferencialmente expressas. A
integração dos dados transcricionais e proteômicos indicou que ainda no início da infecção (6 h), as células de levedura de P. brasiliensis sofreram uma mudança no metabolismo da glicólise para β-oxidação, enzimas desintoxicantes reguladas para se defender contra o estresse oxidativo e repressão da biossíntese de parede. Análises de bioinformática e funcionais também demonstraram que uma serino proteinase foi regulada e secretado in vivo. Pelo o nosso conhecimento, este é o primeiro estudo que descreve a transcrição e dados proteômicos de células de levedura de P. brasiliensis após 6 h de infecção no pulmão de camundongo.
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Comportement biomécanique de la paroi abdominale et de ses composants musculaires : du spécimen isolé au patient / Biomechanical behavior of the abdominal wall and its muscular components : from isolated specimens to patientTran, Doris 10 December 2013 (has links)
Dans le but d’améliorer le traitement des éventrations, cette thèse porte sur la biomécanique de la paroi abdominale. Plusieurs aspects de son comportement ont été étudiés: de la paroi globale aux constituants, de la paroi passive ex vivo sous chargement contrôlé à la paroi active in vivo sous chargement physiologique et enfin de la paroi saine des volontaires non-malades à la paroi lésée des patients en pré-opératoire. Un protocole a été développé pour évaluer la contribution des composants de la paroi abdominale humaine ex vivo à sa réponse mécanique globale. Les spécimens sont sollicités par pression après dissection successive des différents composants. L’analyse par stéréo-corrélation des déformations de la surface interne a montré que la gaine des rectus abdominis joue un rôle important dans la réponse de la paroi abdominale antérieure. Les examens in vivo ont permis de considérer la paroi abdominale in situ de 11 volontaires non-malades et pour la première fois de patients souffrant d’éventration (n=4) en pré-opératoire. Géométrie externe et interne, élasticité des muscles et raideurs locales ont été mesurées lors d’activités physiologiques. Les paramètres mécaniques mesurés sur les patients restent soit dans l’étendue de mesure des valeurs des volontaires non malades, soit sont plus faibles. Dans le futur, l’inclusion d’autres patients et des examens post-opératoires seront poursuivis. On disposera alors de données quantitatives de la paroi abdominale couvrant l’état sain, lésé et réparé. Des modèles numériques pourront être développés. Ils permettront d’estimer l’influence de paramètres mécaniques et géométriques sur le comportement de la paroi abdominale / This thesis focuses on the biomechanics of the abdominal wall in order to help improving the treatment of incisional hernias. Several aspects of its response werestudied: from global wall to components, from passive wall ex vivo subjected to controlled loading to active wall in vivo under physiological loading and finally from intact wall of healthy volunteers to incised wall of herniated patients before repair surgery. A protocol was developed to assess the contribution of the components of the human abdominal wall ex vivo to the global mechanical response. The specimens were loaded by air pressure after dissection of different components. The strain analysis of the internal surface by stereo-correlation highlighted the importance of the rectus abdominis sheath in the response of the anterior abdominal wall. The in vivo examinations considered the entire abdominal wall in situ of 11 healthy volunteers and for the first time herniated patients before surgery (n=4). External and internal geometry, muscle elasticity and local stiffness were measured during various physiological activities. Mechanical parameters measured on patients remain either in the range of values obtained with healthy volunteers or lower. In the future, the inclusion of other patients and post-operative examinations will be conducted. This will provide quantitative data for the healthy, incised and repaired abdominal wall and support the development of numerical models to estimate the influence of mechanical and geometrical parameters on the behavior of the abdominal wall
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Alterations of the Monoaminergic Systems in the Rat Brain by Sustained Administration of Carisbamate and LamotrigineShim, Stacey January 2012 (has links)
Carisbamate (CRS) and lamotrigine (LTG) are anticonvulsants which act mainly on neuronal voltage-gated sodium channels, that have been shown to have antidepressant-like effects in animal models of depression. In vivo electrophysiological recordings were carried out following 2 and 14 days of CRS or LTG administration. Overall firing activity in the dorsal raphe, locus coeruleus and ventral tegmental area were decreased with CRS. Similarly, a decrease in the dorsal raphe was also observed with LTG. Despite these presynaptic decreases in firing activity, both anticonvulsants exhibited significant enhancement of serotonergic transmission in the hippocampus as demonstrated by increased tonic activation of postsynaptic 5-HT1A receptors. This may be attributed to the observed desensitization of the terminal 5-HT1B autoreceptors. This study suggests that the enhanced serotonergic effect may be associated with an antiglutamatergic effect, and may contribute to the antidepressant-like effect of CRS in the forced swim test and the antidepressant properties of LTG.
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The effect of maternal nicotine exposure on the quantity and quality of neonatal rat lung connective tissueDolley, Larry January 1994 (has links)
>Magister Scientiae - MSc / The infants of smoking mothers (compared to non-smoking mothers) have been shown to have a lower birth mass, a lower brain mass, an increased perinatal mortality rate as well as a predisposition to respiratory abnormalities in later life. Evidence suggests that one of the reasons for the latter is abnormal lung structure due to changes in the connective tissue skeleton. This study evaluated the in vivo effects of maternal nicotine exposure (lmg/kg/day subcutaneously - designated the experimental group), which is equivalent to smoking 32 cigarettes per day, on the connective tissue status of the neonatal (7, 14 and 21 day old) wistar rat lung. The control group received sterile saline as a placebo. The specific aspects investigated were: (1) the morphological changes in lung structure and connective tissue (collagen, elastic tissue and reticulin) distribution by means of light microscopy. (2) the quantities of collagen and Emphysema-like morphological changes are present at all ages. The histochemical appearance of collagen is not affected while reticular fibres appear to be abnormal in structure. On day 7 there appears to be no elastic tissue in the nicotine-exposed lung compared to the control lung. This difference is notelastic tissue in the lung. (3) the ultrastructure of the lung connective tissue skeleton by means of scanning electron microscopy. noticeable on days 14 and 21.
Biochemical quantitation indicated that, for the three age groups studied, there was no significant difference in collagen content between experimental and control animals. Elastic tissue was significantly higher in 7 day old experimental lungs than in the control group, contradictory to the results of the histochemical studies. This difference was not significant for 14 and 21 day old lungs Ultrastructural studies of the lung connective tissue skeletons hoed abnormal fibres in the experimental group. Changes included fibre breaks, a beaded appearance of certain fibres and a deficiency in normal fibre arrangement due to the direct or indirect effects of nicotine The effects of nicotine on neonatal rat lung after maternal nicotine exposure is described. The direct mechanisms for these events are still not known but speculation as to this are presented here. Further studies which could explain these mechanisms are also suggested.
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Regional differences in the response of the hamster airway epithelium to elastase: In vivo and In vitro studiesAlonso, Pedro A. January 1994 (has links)
Thesis (Ph.D.)--Boston University / PLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you. / The hamster model of experimental chronic bronchitis comprises a persistent increase in the proportion of bronchial granulated secretory cells after a single intratracheal instillation of elastase. This granulated secretory cell increase, which does not occur in the trachea, has been termed secretory cell metaplasia (SCM). Susceptibility of the bronchial epithelium may be due to a large population of elastase-responsive cells specific to this region. Three dimensional reconstruction of the major form of bronchial secretory cells revealed very little or no rough endoplasmic reticulum (RER), thus demonstrating significant regional heterogeneity since all epithelial secretory cells in the trachea have abundant RER. Animals with bronchial SCM were stimulated with pilocarpine to determine whether the cells subsequent to discharge would re-accumulate granules, thus indicating a permanent phenotypic change. However, bronchial secretory cells failed to discharge at doses equal to and greater than those claimed to be effective in rats. Elastase instilled intratracheally was immuno-localized in the hamster airways to assess the possibility of regional differences in cellular uptake of the enzyme. Elastase was not seen intracellularly in trachea or bronchus suggesting that initiation of bronchial SCM results from a cell surface effect, possibly because of elastase-specific sites on bronchial but not tracheal cells. Tracheal resistance was tested by challenging the epithelial cells in vivo and in vitro with very high doses of elastase. Light and electron microscopy revealed no evidence of a stimulation of the mucus synthetic apparatus, suggesting that tracheal epithelial cells are inherently resistant to proteolytic up-regulation. / 2031-01-01
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Sialic acids: their in vitro and in vivo inhibitation of antibody-antigen agglutinogen reactionsRule, Allyn L. January 1965 (has links)
Thesis (Ph.D.)--Boston University / PLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you. / The in vitro relationship of sialic acids to the A, B, M, N, C, D, and E antigens of the human erythrocyte has been studied by means of the Landsteiner hapten inhibition test with the idea that substances that strongly inhibit anti-D might find practical application in the prevention and treatment of erythroblastosis fetalis. Our results suggest that N-acetyl neuraminic acid (NANA) is a major constituent of the D (Rh0), M, and N agglutinogens, a minor constituent of the A antigen, but is probably not a functional portion of the B, C, and E antigenic structures. [TRUNCATED] / 2031-01-01
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Účinky vybraných flavonolygnanů silymarinu ex vivo na izolované aortě potkana / The ex vivo effects of selected silymarin flavonolygnans on isolated rat aortaSloukgi, Tatiana January 2020 (has links)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Tatiana Sloukgi Supervisor: PharmDr. Jana Pourová, Ph.D. Title of Diploma Thesis: The effect of Silymarin Flavonolignans and their sulfated conjugates on blood vessels ex vivo. Silymarin flavonolignans have recently shown some positive effects on the cardiovascular system. In this work, we studied the vasodilatory effect on rat aorta ex vivo of three silymarin conjugates, silybin A-20-sulfate, silybin B-20-sulfate and 2,3- dehydrosilychristin-19-O-sulfate, and one parent flavonolignan 2,3- dehydrosilychristin. For each substance, a concentration response curve was created and the concentration that produces 50% of maximum relaxation was determined (EC50). All substances exerted very low or no vasodilatory activity. Finally, we focused on the mechanism of action of silybin A. We tested whether its vasorelaxant activity depends on the presence of intact endothelium. The vasorelaxant effect of silybin A on isolated rat aorta ex vivo was clearly endothelium-dependent.
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