Dissertations / Theses on the topic 'Granuloma'

Die Wegener'sche Granulomatose (WG) ist eine Autoimmunerkrankung, die sich typischerweise als chronische Entzündung des oberen Respirationstraktes, Vaskulitis und Glomerulonephritis manifestiert. WG gehört zur Gruppe der sog. “pauci-immunen” Vaskulitiden, die mit Anti-Neutrophilen-Antikörpern (ANCA, anti neutrophil cytoplasmic antibody) assoziiert sind. Mit Hilfe der indirekten Immunfluoreszenz lassen sich ANCA im Serum der meisten WG-Patienten nachweisen. Die mit WG assoziierten sog. “klassischen” ANCA (c-ANCA) erkennen spezifisch Konformationsepitope der Serinprotease Proteinase 3 (PR3), die in den azurophilen Granula neutrophiler Granulozyten gespeichert wird. Die enge Korrelation PR3-spezifischer Antikörperspiegel mit dem Krankheitsverlauf läßt vermuten, daß sie bei der Pathogenese eine zentrale Rolle spielen könnten. Diese Hypothese wird von Daten aus in vitro Experimenten gestützt: werden Zytokin-stimulierte neutrophile Granulozyten mit Patientenserum oder isolierten c-ANCA inkubiert, erfolgt eine Aktivierung der Neutrophilen, die sich durch Degranulation und Freisetzung von Sauerstoffradikalen äußert. c-ANCA können so indirekt - aber vermutlich auch direkt - zur Endothelschädigung beitragen. Jedoch konnte bisher kein direkter Beweis des pathogenen Potentials von c-ANCA am Tiermodell erbracht werden. Um die Wirkung von c-ANCA an einem Mausmodell zu testen, war zunächst ein murines Maus-PR3- (mPR3) spezifisches Antiserum notwendig. Da humane c-ANCA nicht mit mPR3 kreuzreagieren, wurde für die Immunisierung von PR3/Neutrophilen-Elastase (NE)-defizienten Mäusen ein mPR3-Zymogen rekombinant in E. coli als Einschlußkörpermaterial (IB, inclusion bodies) hergestellt. Nach Renaturierung des IB-Materials in vitro wurde mit der Dipeptidylaminopeptidase Cathepsin C das N-terminale Propeptid abgespalten. Das gewonnene Enzym besaß die für PR3 und NE spezifische katalytische Aktivität, die durch den physiologischen Inhibitor humaner PR3, a1-Antitrypsin, inhibiert werden konnte. Es ist daher anzunehmen, daß das gewonnene rekombinante Material die korrekte Konformation durch Renaturierung in vitro erhalten hatte. Um nun die pathologische Wirkung von Anti-rmPR3-Antikörpern zu testen, wurden PR3/NE-defiziente Mäuse mit dem rekombinanten Zymogen (pro-rmPR3) oder der N-terminal prozessierten Form (rmPR3) immunisiert. Die Spezifität der gewonnenen Antiseren wurde durch Festphasenimmunoassay, Western Blotting und indirekte Immunfluoreszenzfärbung überprüft. Weiterhin konnte fluoreszenzzytometrisch die Bindung von Anti-mPR3-IgG an die Plasmamembran Zytokin-stimulierter Granulozyten nachgewiesen werden. Die hergestellten Antiseren erfüllten somit die für c-ANCA-positive Seren von WG-Patienten typischen Eigenschaften hinsichtlich der Antigenspezifität. Wenn c-ANCA alleine hinreichend für die Induktion der für WG charakteristischen Symptome sind, sollte der Antiserumtransfer auf Wildtyp-Mäuse WG-ähnliche Symptome in den Rezipienten hervorrufen. Nach wiederholter intravenöser Injektion von Serum pro-rmPR3-immunisierter Mäuse ließ sich ein signifikanter Antikörperspiegel bei Verdünnungen von 1:2000 über den gesamten Behandlungszeitraum von 10 Wochen in den Rezipienten nachweisen. Der anschließende histologische Befund von Niere und Lunge ergab jedoch keine pathologischen Veränderungen. Dieses Ergebnis legt nahe, daß c-ANCA alleine keine Krankheitssymptome hervorrufen. In dem gegenwärtigen Modell für c-ANCA-vermittelte Vaskulitis entfaltet sich die pathogene Wirkung von c-ANCA vor allem dann, wenn neutrophile Granulozyten zusätzlich durch proinflammatorische Zytokine wie Tumornekrosefaktor alpha (TNF alpha) stimuliert werden. Erst die Stimulation der Granulozyten ermöglicht die Bindung von c-ANCA an die Plasmamembran und deren anschließende Aktivierung. Deshalb wurde dieses Modell, das vorwiegend aus Ergebnissen von Experimenten in vitro abgeleitet ist, auf ein lokales Entzündungsmodell der Maus übertragen: Durch wiederholte Injektion von TNF alpha in die Haut wurde eine leichte Entzündungsreaktion ausgelöst. Diese Entzündungsreaktion ließ sich schließlich durch intravenöse Verabreichung von pro-rmPR3- oder rmPR3-Antiserum verstärken. Dieser Befund ist ein wichtiger Beweis für die verbreitete Ansicht, daß c-ANCA nicht nur ein Epihänomen der WG darstellen, sondern selbst ein pathogenes Potential besitzen. Im zweiten Teil der vorliegenden Arbeit wurde die Beteiligung der beiden Serinproteasen NE und PR3 an der Entstehung inflammatorischer Prozesse untersucht. Im Hinblick auf die bei der Pathogenese der WG beteiligten Mechanismen könnten PR3 und NE eine wichtige Rolle spielen. PR3 und NE können Proteine der extrazellulären Matrix abbauen, Apoptose in Endothelzellen induzieren und sind an der Regulation entzündlicher Prozesse über verschiedene Wirkmechanismen beteiligt. Um quantitative Unterschiede bei Entzündungsreaktionen zwischen NE/PR3-defizienten Mäusen und kongenen Wildtyp-Tieren zu untersuchen, wurde als Modell einer Typ III Hypersensitivitätsreaktion eine lokale passive Arthus-Reaktion induziert. Wildtyp-Mäuse entwickelten dabei eine deutlich stärkere lokale Entzündung als NE/PR3-defiziente Mäuse. Weitere Studien sind nötig um die Frage zu klären, ob eine der beiden Serinproteasen alleine oder in Kooperation mit der anderen diesen Phänotyp hervorruft. Für einen direkten synergistischen Effekt sprechen indes die Ergebnisse eines in vitro Experiments mit pro-rmPR3 und humaner NE: Bei Inkubation von pro-rmPR3 mit hNE wurde eine Spaltung des Proenzyms beobachtet, die mit einer Verstärkung der enzymatischen Bruttoaktivität einherging. Die physiologische Relevanz dieser Beobachtung muß allerdings noch geklärt werden. Die Ergebnisse der vorliegenden Arbeit stehen im Einklang mit den neuesten Erkenntnissen über die Rolle der PR3 bei der Wegener’schen Granulomatose: PR3 dürfte sowohl aufgrund pleiotroper Effekte auf entzündliche Reaktionen als auch wegen seiner lytischen Eigenschaften zur Gewebeschädigung beitragen. Darüberhinaus konnte eine pathogene Wirkung von mPR3-spezifischen Antikörpern in der Maus nachgewiesen werden
Wegener's granulomatosis (WG) is an autoimmune disorder typically characterized by chronic inflammmation of the upper respiratory tract, vasculitis and glomerulonephritis. WG belongs to the group of anti-neutrophil cytoplasmic autoantibody (ANCA) associated vasculitides with no or very few immune complex deposits (“pauci-immune”) in affected organs. “Classic” ANCA (c-ANCA) that produce a cytoplasmic staining pattern on neutrophils are a specific seromarker for this disease entity. They recognize conformational epitopes of proteinase 3 (PR3) from azurophil granules of neutrophil granulocytes. The correlation of PR3-specific antibody titers and disease course suggests that they are an important pathogenic factor for this autoimmune disease. The hypothesis is supported by in vitro experiments demonstrating an interaction of c-ANCA with cytokine primed neutrophils resulting in full activation manifested by degranulation and respiratory burst. It has been shown that c-ANCA can also directly induce the loss of endothelial barrier functions. However, direct evidence for the pathogenic potential of c-ANCA in vivo has not been published yet. The central aim of this study is to clarify the role of c-ANCA in WG. Since antibodies directed against human PR3 do not crossreact with the murine homolog, mPR3-specific antibodies were necessary to provide direct proof for c-ANCA mediated tisssue damage in a murine disease model. Hence, sufficient amounts of recombinant murine PR3 (mPR3) were necessary to generate mPR3 specific murine antibodies. Several attempts have been made to produce recombinant PR3 in prokaryotic and eukaryotic host systems. But conformational epitopes recognized by c-ANCA are not well preserved on recombinant PR3 derived from E. coli, P. pastoris, or baculovirus-infected insect cells described in the literature so far. While recombinant human PR3 expressed in eukaryotes is well recognized by c-ANCA, the yield of both active human and murine PR3 generated in eukaryotic expression systems is very low. To obtain sufficient amounts of correctly folded recombinant murine proteinase 3 (rmPR3) for multiple immunizations of PR3/NE-deficient mice, rmPR3 was produced as inclusion body material in E. coli as a catalytically inactive precursor molecule. After in vitro refolding the N-terminal propeptide was removed by limited proteolysis with the dipeptidylaminopeptidase cathepsin C yielding catalytically active enzyme. Due to its catalytic activity that could be inhibited by the physiologic inhibitor of human PR3, α1-antitrypsin, but not by secretory leukocyte protease inhibitor (SLPI), the recombinant material was assumed to harbour the correct conformation after refolding. To test, if anti-rmPR3 antibodies were sufficient to induce symptoms characteristic for WG, anti-PR3 antiserum was generated by immunization of PR3/neutrophil elastase (NE)-deficient mice with recombinant, refolded mPR3 or its zymogen. Specificity of the obtained sera was confirmed by ELISA, Western Blotting and indirect immunofluorescence. Moreover, antisera bound to the membranes of primed murine neutrophils as determined by flow cytometry. The generated antisera thus fulfilled the criteria defining c-ANCA positive sera of WG patients with respect to antigen specificity. If anti-PR3 antibodies are sufficient to induce symptoms characteristic for WG, transfer of antiserum to wildtype mice should induce vasculitis and/or glomerulonephritis. By repetitive intravenous injection of antiserum from immunized mice a persisting antibody titer was generated in naïve wild type mice over a treatment period of 10 weeks. Lung and kidney of these mice were analyzed histologically but neither granuloma or vasculitis were found in the lungs nor glomerulonephritis or vasculitis was observed in the kidneys. This result suggests that c-ANCA alone are not sufficient to induce WG symptoms in the mouse. Our initial observations were not surprising since the current hypothetical concept of c-ANCA-induced vasculitis implies that a primary inflammatory stimulus provided by cytokines like TNF alpha is required for the target antigen to be expressed on the plasma membrane of neutrophils. Exposure of the target antigen enables the binding of c-ANCA and subsequently triggers neutrophil activation. Consequently, this model, that was primarily deduced from in vitro experiments, was adapted to a model of local inflammation in the mouse: A mild inflammation was induced by repetitive local injection of TNF alpha into the skin. This inflammatory reaction increased significantly in the presence of rmPR3-antibodies. Our experimental data thus confirm the current concept that ascribes a pathogenic potential to c-ANCA. A second aspect adressed in this work is the contribution of the two neutrophil serine proteases NE and PR3 to the generation of inflammatory processes. With respect to the mechanisms involved in the pathogenesis of WG, NE and PR3 may be of particular importance due to their ability to degrade extracellular matrix proteins, induction of apoptosis in endothelial cells, and the regulation of inflammatory processes by a variety of mechanisms. A local reverse passive Arthus reaction (RPA) was chosen as a model of a type III hypersensitivity reaction to reveal quantitative differences of inflammation in PR3/NE-deficient mice and congenic wild type mice. Wild type mice reacted significantly stronger than PR3/NE-deficient mice as determined by examination of local edema and hemorrhage intensity. It remains to be determined if the observed phenotype in vivo reveals a concerted effect of both serine proteases or if deficiency of one of the proteases alone accounts already for this phenotype. Experimental data presented in this work are consistent with the hypothesis that PR3 and NE may directly interact: When recombinant mPR3 was incubated with hNE, a cleavage of rmPR3 was observed that is apparently associated with changes in enzymatic activity. The physiologic relevance of this finding has to be defined in further studies. In summary, this work adds to the current understanding of the role of PR3 in WG: PR3 is not only the relevant autoantigen whose interaction with c-ANCA contributes to the fatal outcome of the disease but also contributes together with neutrophil elastase to tissue damage by its lytic activity and pleiotropic effects on inflammatory reactions

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Os Granulomas Dentários e Cistos Radiculares representam lesões periapicais crônicas que, frequentemente, acometem os ossos maxilares. As Células de Langerhans são células dendríticas, responsáveis pela apresentação de antígenos aos Linfócitos T, que desempenham importante função nos tecidos epiteliais, bem como na patogênese das lesões periapicais. O presente estudo analisou a expressão das Células de Langerhans, através da técnica imuno-histoquímica para o marcador CD1a em 18 casos de Granuloma Dentário (GD) e 26 casos de Cisto Radicular (CR). Essas células dendríticas foram observadas em 11,1% dos Granulomas Dentários e em 69,2% dos Cistos Radiculares, mostrando correlação estatisticamente significante (p-valor=0,000. Teste de Fisher). Nos Cistos Radiculares, as CLs exibiram tanto a forma arredondada quanto a dendrítica, em todas as camadas epiteliais. Já nos Granulomas Dentários, as CLs foram vistas apenas no tecido de granulação com densidade discreta de marcação. Apesar de termos encontrado uma correlação entre densidade de marcação e espessura de epitélio, bem como entre imunomarcação e intensidade inflamatória, não foi observada representatividade estatística entre essas correlações. Dos resultados obtidos conclui-se que as Células de Langerhans parecem influenciar na imunopatogênese das lesões periapicais aqui estudadas, principalmente nos Cistos Radiculares.
Salvador

IFN- plays an important role in the pathogenesis of periapical lesions, and the methylation of IFNG has been associated with transcriptional inactivation. The purpose of the present study was to investigate IFNG promoter methylation in association with gene transcription and protein levels in periapical granulomas and radicular cysts. Methylation-specific (MSP) PCR was used to assess the DNA methylation pattern of the IFNG gene in 16 periapical granuloma and 13 radicular cyst samples. The transcription levels of IFNG mRNA were verified by quantitative real time PCR (qPCR), and protein expression was evaluated by immunohistochemistry. All the periapical lesion samples exhibited partial or total methylation of the IFNG gene. In addition, an increased methylation profile was found in radicular cysts compared to periapical granulomas. Increased IFNG mRNA expression was observed in the partially methylated periapical lesion samples relative to the samples that were completely methylated. The present study provides the first evidence of the possible impact of IFNG methylation on IFNG transcription in periapical lesions.
O IFN- apresenta importante função na patogênese das lesões periapicais e a metilação do gene IFNG tem sido associada à inativação da transcrição. O objetivo deste estudo foi investigar a metilação da região promotora do IFNG e a associação com a transcrição do gene e com os níveis de proteína no granuloma periapical e no cisto radicular. O PCR específico para metilação (MSP Methylation Specific PCR) foi usado para avaliar o padrão de metilação do DNA do gene IFNG em 16 amostras de granuloma periapical e em 13 de cisto radicular. Os níveis de transcrição do RNAm do IFNG foram verificados pelo PCR em Tempo Real (qPCR) e a expressão da proteína foi avaliada por meio da imunoistoquímica. Todas as amostras de lesão periapical exibiram metilação total ou parcial do gene IFNG. Além disso, um aumento no perfil de metilação foi encontrado nos cistos radiculares em relação aos granulomas periapicais. Observou-se um aumento na expressão do RNAm do IFNG em amostras de lesão periapical parcialmente metiladas em relação àquelas totalmente metiladas. Este estudo apresenta a primeira evidência do possível impacto da metilação do IFNG na transcrição do IFNG em lesões periapicais.

Orientador: Antonio Condino Neto
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas
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Resumo: As infecções de repetição são queixas frequentes no consultório pediátrico. Apesar das caracteristicas próprias de infecção recorrente nesta faixa etária, o Pediatra deve estar atento para a presença de defeitos imunológicos como fator determinante dos quadros infecciosos de repetição. Entre as imunodeticiências primárias, a Doença Granulomatosa Crônica (DGC) é causada por alteração no sistema NADPH oxidase das células fagocíticas. O objetivo deste estudo foi avaliar aspectos clínicos de 29 pacientes encaminhados a um laboratório especializado, com suspeita de defeito de fagócitos a nível do sistema NADPH oxidase. Foram realizados os testes de NBT e ânion superóxido e os pacientes foram divididos em dois grupos: grupo I para os pacientes com alteração no sistema NADPH oxidase e grupo TI para os pacientes que não apresentaram alteração neste sistema. A ocorrência de infecção urinária esteve associada ao grupo em que não se confirmou o diagnóstido de DGC. A presença de história familiar de infecção de repetição apresentou forte associação ao grupo com diagnóstico DGC. Outros dados da história clínica não apresentaram diferenças estatisticamente signifivativa entre os 2 grupos. No grupo de pacientes com DGC observou-se a correlação genótipo-fenótipo descrita na literatura, onde pacientes com a forma ligada ao sexo apresentam início dos sintomas mais precocemente e quadros infecciosos mais graves
Abstract: Recurrent infections are ftequently referred to the Pediatrician' s office. As the irnrnune system continues to develop after birth and will be completed just after childhood, infants tend to present more infections. Besides this particular characteristic, Pediatricians should be aware to some abnormal conditions, the primary immunodeficiencies. Among the primary immunodeficiencies, Chronic Granulomatous Disease is the result of a defect in any of the proteins of the NADPH oxidase system of human phagocytic cells. The aim of this study was to evaluate the clinical aspects of 29 patients that were referred to a specialized laboratory, being suspected of having a defect in the NADPH oxidase system. It was performed the NBT slide test and superoxide dosage. Patients were divided into two groups: group I for patients with defect in this system and group TI for patients without defects in this system. Recurrent urinary tract infections were associated with group TI. A history of recurrent infections in the family was strongly associated to group I. Among CGD patients it was observed the genotype-phenotype correlation presented in the literature in wich X-CGD is associated with an earlier onset of clinical manifestations and more severe infections
Mestrado
Pediatria
Mestre em Saude da Criança e do Adolescente

A esquistossomose, doença causada pelo parasito Schistosoma mansoni, é um problema de saúde mundial. A paracoccidioidomicose (PCM) é uma micose sistêmica humana, causada pelo fungo Paracoccidioides brasiliensis, e pode coinfectar o hospedeiro portador de S. mansoni. Na confecção por S. mansoni, o sistema imune do hospedeiro pode estar suprimido, o que acarreta uma defesa insuficiente para eliminação de outros microrganismos. Até o momento, existem poucos trabalhos que descrevem a evolução da esquistossomose e da paracoccidioidomicose em casos de coinfecção. Além disto, devido a correntes migratórias, indivíduos de áreas endêmicas para esquistossomose podem se instalar em áreas endêmicas para PCM e vice versa. Considerando estes antecedentes, este trabalho teve como objetivo analisar aspectos histopatológicos e imunológicos do desenvolvimento do granuloma esquistossomótico durante a coinfecção Para isto, camundongos Swiss fêmeas foram separados em 4 grupos para fase aguda e 4 grupos para fase crônica, com 10 animais em cada: controle não infectado - CNI, infectado com S. mansoni - Sm, infectado com P. brasiliensis - Pb, coinfectado - Sm+Pb, ambos em fase crônica, e coinfectados sendo Sm fase crônica e Pb fase aguda - Sm(c)+Pb(a). Os camundongos foram necropsiados após 50 (fase aguda) e 120 dias (fase crônica) para análise quanto ao peso, sobrevida, desenvolvimento do granuloma e produção de citocinas. Em relação ao peso, observou-se que não houve alterações no peso dos camundongo após 50 dias. Já para a fase crônica, foi observado ganho significativo de peso no grupo Pb. Para a sobrevida, em fase aguda, observou-se 10% de óbitos no grupo Sm. Já para fase crônica, 50% de óbitos para o grupo Sm, 60% do grupo Sm+Pb e 50% do grupo Sm(c)+Pb(a) foram observados ao final do experimento. A ocorrência da coinfecção foi evidenciada a partir da visualização de granulomas no fígado e no pulmão, e a presença de fungo no epiplon. A contagem de granulomas no fígado (fase aguda e crônica) e pulmão (fase crônica), não apresentou diferenças significativas. Em relação ao diâmetro dos granulomas no fígado em fase aguda, estes foram significativamente maiores no grupo Sm (0,08±0,05 mm²) quando comparado ao grupo Sm+Pb (0,06±0,05mm²). Na fase crônica, o diâmetro foi significativamente maior quando comparados os grupos Sm+Pb (0,03±0,01mm²) ao Sm(c)+Pb(a) (0,02±0,01mm²). Em relação ao pulmão, houve diferenças significativas elevadas quando comparados os grupos Sm (0,01±0,009mm²) com Sm+Pb (0,03±0,01mm²) e Sm(C)+Pb(A) (0,03±0,02mm²). Foram evidenciadas células mononucleares e polimorfonucleares nos granulomas de fígado e pulmão em todos os grupos estudados, tanto em fase aguda e crônica. Em fase aguda, a contagem de eosinófilos e neutrófilos não obteve diferenças significativas em relação aos tipos celulares, apesar de um número maior de neutrófilos serem observados no grupo Sm+Pb (7,82±5,95). Na fase crônica, verificamos que no grupo Sm houve um predomínio significativo de eosinófilos (17,34±6,71), enquanto nos grupos Sm+Pb e Sm(c)+Pb(a) o predomínio foi de neutrófilos (18,82±8,18 e 12,72±4,44, respectivamente). No pulmão, não houve diferença em relação ao número de eosinófilos presentes nos granulomas, já o número de neutrófilos no grupo Sm(c)+Pb(a) (2,66±2,71) que foi significativamente menor em comparação aos grupos Sm (9,62±7,76) e Sm+Pb (8,06±5,40). Os resultados da análise de citocinas nos diferentes grupos mostram um aumento significativo de IFN-γ nos grupos Sm (8,05±3,52ng/ml) e Pb (10,40±3,549ng/ml) em relação ao CNI (1,14±0,48 ng/ml). A citocina IL-2 apresentou níveis significativamente mais elevados nos grupos Sm (6,71±1,50ng/ml), Pb (10,40±3,54ng/ml) e Sm+Pb (7,32±2,93ng/ml), quando comparados ao grupo CNI (2,01± 0,39ng/ml). Para IL-4, houve diferenças significativas elevadas entre os grupo Sm (10,45±3,67ng/ml) quando comparados ao grupo CNI (0,85±0,15ng/ml). Para IL-5, diferenças significativas foram encontradas nos grupos Sm (3,91±0,61ng/ml), Pb (3,98±1,76 ng/ml) e Sm+Pb (4,28±2,22ng/ml), quando comparados ao CNI (0,67±0,21ng/ml). Em fase crônica, foram observados níveis significativamente elevados de IFN-γ no grupo Sm (10,45± 3,67ng/ml) comparado ao CNI (1,14±0,48ng/ml). Para IL-2, houve diferenças significativas elevadas nos grupos Sm (7,90±3,16ng/ml) e Pb (7,54±1,41ng/ml), quando comparados ao CNI (2,01±0,39ng/ml). Para IL-4 e IL-5, diferenças significativas elevadas foram observadas para o grupo Sm (8,46±3,71ng/ml e 4,47±1,88ng/ml, respectivamente) quando comparados ao CNI (0,85±0,15ng/ml e 0,67± 0,21 ng/ml, respectivamente). Em relação a MIP-2, não foram observados diferenças significativas entre os grupos, tanto para fase aguda e crônica. Sendo assim, os dados sugerem que a coinfecção por P. brasiliensis pode influenciar na composição celular dos granulomas esquistossomóticos e nos níveis de citocinas.
Schistosomiasis, also known as snail fever, a disease caused by the parasite Schistosoma mansoni, is a worldwide health issue. Paracoccidioidomycosis (PCM) is a human systemic mycosis caused by the fungus Paracoccidioides brasiliensis and it may coinfect the bearing host of S. mansoni. In the coinfection by S. mansoni, the host immune system may be suppressed which results in an insufficient defense in order to obliterate other microorganisms. There are few studies so far describing the evolution of schistosomiasis and paracoccidioidomycosis in the case of coinfection. Furthermore, due to migration, people from schistosomiasis endemic areas may settle down in PCM endemic areas and vice versa. Considering the given background, this study had as main goal the analysis of histopathological and immunological aspects of the development of schistosomotic granuloma under the coinfection. Thereunto, Swiss female mice were separated in four groups for acute phase and five groups for chronic phase containing ten animals each: control not infected - CNI, infected with S. mansoni - Sm, infected with P. brasiliensis - Pb, coinfected – Sm+Pb, both in chronic phase, and coinfected Sm in chronic phase and Pb in acute phase – Sm(c)+Pb(a). The mice were necropsied after 50 days (acute phase) and 120 days (chronic phase) for analysis concerning their weight, survival rate, granuloma development and cytokine production. Regarding the weight, it was observed the Pb group had gained significantly weight (26,23±5,36g). Concerning the survival rate, in acute phase, it was observed 10% of deaths in the Sm group; as for the chronic phase, 50% of deceased for Sm group, 60% for Sm+Pb group and 50% for Sm(c)+Pb(a) by the end of the experiment. The coinfection occurrence was demonstrated by granuloma visualization in the liver and lung, and presence of the fungus in the omentum. The granuloma count in the liver (acute and chronic phases) and lung (chronic phase) did not display any meaningful differences. Regarding the diameter of the granulomas in the liver in acute phase, it was meaningfully bigger than Sm group (0,08±0,05 mm²) when compared to Sm+Pb group (0,06±0,05mm²). In the chronic phase, the diameter was quite bigger when compared to Sm+Pb (0,03±0,01mm²) and Sm(c)+Pb(a) (0,02±0,01mm²) groups. Toward the lung , there were high significant differences when compared Sm groups (0.01 ± 0,009mm²) with Sm+Pb (0.03 ± 0,01mm² ) and Sm(C)+Pb(A) (0.03 ± 0,02mm²) .Mononuclear and polymorphnuclear cells of liver and lung granulomas were evidenced in all groups both in acute and chronic phases. In acute phase, the eosinophil and neutrophils counts did not have meaningful distinctions regarding the cell types, although a bigger number of neutrophils were observed in Sm+Pb group (7,82±5,95). In chronic phase, it was verified a distinguished prevalence of eosinophils (17,34±6,71) in Sm group, whilst Sm+Pb and Sm(c)+Pb(a) groups neutrophils prevailed (18,82±8,18 e 12,72±4,44, respectively). In the lung, there were not any differences in regard of eosinophil number in granulomas, while the number of neutrophils in Sm(c)+Pb(a) group (2,66±2,71) was really smaller than Sm (9,62±7,76) and Sm+Pb (8,06±5,40) groups. The results of cytokine analysis from those different groups display a distinguished augmentation of IFN-γ in Sm (8,05±3,52ng/ml) and Pb (10,40±3,549ng/ml) groups regarding the CNI (1,14±0,48 ng/ml), however this increase observed in Sm+Pb group (4,96±1,73ng/ml) wasn’t meaningful at all. The cytokine IL-2 displayed notably higher levels in Sm (6,71±1,50ng/ml), Pb (10,40 3,54±ng/ml) and Sm+Pb groups (7,32±2,93ng/ml) when compared to CNI (2,01± 0,39ng/ml). As for IL-4, there were expressive high differences between Sm (10,45±3,67ng/ml), Pb (3,98±1,76 ng/ml) and Sm+Pb groups (4,28±2,22ng/ml) regarding CNI (0,67±0,21ng/ml). For IL-5, significant differences were found in groups Sm (3.91 ± 0,61ng / ml), Pb (3.98 ± 1.76 ng / ml) and Sm + Pb (4.28 ± 2,22ng / ml) compared to the CNI (0.67 ± 0,21ng / ml). In chronic phase, it was observed denoting high levels of IFN-γ in Sm group (10,45± 3,67ng/ml) when compared to CNI (1,14±0,48ng/ml). Concerning IL-2, there were compelling differences from Sm (7,90±3,16ng/ml) and Pb groups (7,54±1,41ng/ml), in regard of CNI (2,01±0,39ng/ml). As for IL-4 and IL-5, expressively high distinctions were observed in Sm (8,46±3,71ng/ml and 4,47±1,88ng/ml, respectively) concerning CNI (0,85±0,15ng/ml and 0,67± 0,21 ng/ml, respectively). Regarding MIP-2, it was not observed any important differences among the groups as for the acute or chronic phases, nevertheless these display higher levels in acute phase rather than in chronic phase. Thus, the data suggest the coinfection by P. brasiliensis may affect the cellular composition from schistosomotic granulomas and cytokine levels.

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CAPES
A inflamação granulomatosa na esquistossomose mansônica acontece quando o ovo fica retido no tecido vivo e não consegue amadurecer ou ser expulso, promovendo alterações patológicas severas no indivíduo infectado. O objetivo deste trabalho foi avaliar quantitativamente as lesões granulomatosas de tecido hepático de camundongos infectados com esquistossomose mansônica, através do uso de lectinas conjugadas a um composto quimiluminescente, o éster de acridina (EA). As lectinas Concanavalina A (Con A), Wheat Germ Agglutin (WGA) e Sambucus Nigra Agglutin (SNA) foram conjugadas ao éster de acridina e incubadas no tecido hepático de camundongos esquistossomóticos. A quimiluminescência foi expressa em Unidades Relativas de Luz (RLU). Fragmentos do tecido hepático infectado e de tecido normal foram incubados com ConA-EA, WGA-EA e SNA-EA e ficou evidenciado que houve um aumento da expressão de α-D-glicose/manose e N-acetilglicosamina nos tecidos infectados em relação ao tecido normal, enquanto que não houve significativa diferença entre os valores de α-NeuNAc-[2→6]-Gal/GalNAc dos fígados infectados e não infectados. Assim, a histoquímica quimiluminescente mostrou-se uma ferramenta eficaz na avaliação de mudanças patológicas causadas por esquistossomose mansônica.

Coccidioidomycosis is a growing problem in the United States. There is still an incomplete understanding of the immunological response associated with human coccidioido-mycosis, although previous studies have indicated that cell-mediated immunity (CMI) is critical in the control of this disease. We have examined necrotizing pulmonary coccidioidal granulomata using immunohistochemical staining for lymphocyte subsets and for the cytokines interleukin-10 (IL-10) and gamma interferon (IFN-?). Discrete perigranulomatous lymphocytic clusters were identified containing roughly equal numbers of T (CD3+) and B (CD20+) lymphocytes. While the number of cells expressing IL-10 was similar in the mantle and in the perigranulomatous clusters, there were significantly more cells expressing IFN-? in the mantle compared to the clusters (P = 0.037). Confocal microscopy revealed that CD4+ T lymphocytes and B lymphocytes are associated with IL-10 production. CD4+CD25+ T lymphocytes were also identified in the perigranulomatous clusters but were not associated with IL-10 production. This is the first report noting perigranulomatous lymphocyte clusters and IL-10 in association with human coccidioidal granulomata, suggesting a down-regulation of cellular immune response.IL-12 is critical in driving a T helper type 1 (Th1) immune response. To understand the mechanism associated with hyporesponsiveness to IL-12 stimulation seen in some anergic patients with disseminated coccidioidomycosis, IL-12 receptor expression and function were determined in PBMCs from immune and nonimmune healthy donors. By using a relative quantitative RT-PCR technique, we found that IL-12R?1 is constitutively expressed in PBMCs and is equally up-regulated by coccidioidal antigen preparation T27K in both immune and nonimmune donors. On the other hand, the IL-12R?2 expression level is increased by T27K only in PBMCs from immune but not nonimmune donors. In addition, the increased IL-12R?2 expression in immune PBMCs is correlated with Stat4 activation and the induction of IFN-? production by IL-12. These data suggest that the IL-12R?2 expression and signal transduction is functional in host immunity against Coccidioides infection. Subsequent research, which involved the stimulation of lymphocytes with autologous dendritic cells (DCs) pulsed with T27K, revealed that the up-regulation of IL-12R?2 expression and signal transduction is associated with the induction of IFN-? production by DCs pulsed with T27K.

Este estudo avaliou pacientes portadores de cromoblastomicose, sendo feita uma categorização clínica em dois grupos, de acordo com a forma clínica da lesão dermatológica: o primeiro composto por indivíduos com lesões elevadas e o segundo formado por casos com lesões planas, com biópsias realizadas antes e depois dos tratamentos. Através de critérios semiquantitativos, os elementos anatomopatológicos foram analisados, constatando-se a presença do padrão granulomatoso chamado granuloma micótico misto organizado, com baixa intensidade em lesões planas e alta em elevadas. Evidenciou-se que pacientes com lesões planas evoluíram bem clinicamente, com exames micológicos negativos e sem recidivas (bons respondedores), porém aqueles com lesões elevadas apresentaram evolução clínica desfavorável, com exames micológicos positivos e doença recidivante ou recalcitrante, apesar dos tratamentos recebidos (maus respondedores). E, entre os componentes histopatológicos reacionais à invasão fúngica, verificou-se associação significativa para fibrose, podendo levar a um pior prognóstico. Estes achados clínicos e histopatológicos conferem a esta patologia uma idéia de polaridade, à guisa dos fenômenos das formas polares da hanseníase, da fagocitose, e dos agentes de baixa virulência. Os autores propõem uma classificação morfológica dos granulomas da CBM em dois tipos polares: granuloma micótico misto organizado (GMMO) polar com alta intensidade dos elementos celulares (que corresponderia ao polar não tuberculóide) e GMMO polar com baixa intensidade (que corresponderia ao polar tuberculóide).
Patients with chromoblastomycosis were studied and dichotomized into two groups according to the clinical lesions: flat and elevated ones which were biopsied before and after treatments. Histopathological structures underwent through semiquantitatively analisys evidencing the mixed organized mycotic granuloma with low intensity of histopathological elements in flat lesions and high in elevated ones. Flat lesions have improved clinically with negative micological studies (good responders) while elevated lesions’ patients did not (bad responders). It was found significant association between evidence of fungus and fibrosis with a poor prognosis. Clinical and histopathological findings suggest a polarity concept to this disease based on the fenomenal polar forms of hanseniasis, fagocitosis and the low virulence agents. The authors proposed a morphological classification of the chromoblastomycosis’ granulomas into two polar types: polar mixed organized mycotic granuloma (MOMG) with high intensity of the cellular elements (like the polar non tuberculoid type) and the polar MOMG with low intensity (like the polar tuberculoid type).

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CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
A esquistossomíase é considerada uma das doenças tropicais negligenciada mais significativa no mundo. Sendo que a presença de apenas um medicamento para o tratamento da infecção leva a busca por novos compostos esquistossomicidas, utilizando os produtos naturais como uma das principais fontes destas novas moléculas. Neste sentido, a lignana Arctiina extraída da espécie Arctium lappa, cujas funções anti-inflamatórias e antiproliferativas já foram descritas na literatura, se tornou alvo do nosso estudo. O nosso propósito foi pesquisar a sua ação esquistossomicida através de testes in vitro e em modelo murino. A substância foi utilizada nas concentrações de 30, 60, 120 e 240 µg/mL nos ensaios in vitro. Após o período de incubação, em nenhuma das concentrações, a molécula foi capaz de promover modificação na viabilidade do parasito em cultura quando comparado ao grupo controle. Após a administração por via intraperitoneal, para verificar a presença da substância no plasma murino, foi realizada uma análise cromatográfica. A análise da amostra de arctiina pura, diluída em metanol, e diluída em plasma murino não tratado mostrou um pico no cromatograma medido a 254 nm, com retenção de 5 minutos. A amostra de plasma animal coletada após uma hora de tratamento com arctiina, sob as mesmas condições experimentais, revelou um pico semelhante ao da amostra pura, confirmando que a arctiina está disponível no plasma após administração. Os testes in vivo, foram realizados em camundongos fêmeas da linhagem Swiis que receberam por via intraperitoneal duas dosagens de arctiina (50 mg/kg), sendo a primeira administrada 20 dias após a infecção e a segunda após duas semanas. Nos parâmetros analisados: peso hepático, leucometria global, redução da carga parasitária e alteração no oograma, não foi verificado nenhuma alteração significativa em relação aos parâmetros encontrados no grupo controle infectado, tratado com praziquantel (200 mg/kg) e Dimetilsulfóxido (0,5%). O resultado mais promissor foi uma redução das médias das áreas dos granulomas, a administração da arctiina provocou uma redução em torno de 20% em comparação com o controle infectado. Mais estudos devem ser realizados a fim de verificar o possível mecanismo de atuação sobre os componentes inflamatórios presentes na formação do granuloma.
Schistosomiasis is one of the most significant neglected tropical diseases in the world. Only one drug is currently available for the treatment and control of schistosomiasis, therefore there is an urgent need for the development of new schistosomicide compounds, being natural products an important source of these molecules. Hence, in this work we studied the lignan arctiin extracted from Arctium lappa species, whose anti-inflammatory and antiproliferative functions have been previously described. Our aim was to investigate in vitro and in vivo its schistosomicidal activity. We tested the compound in vitro at the follow concentrations 30, 60, 120 and 240 ug/ml. There was no difference in all tested concentrations in the viability of the parasite in the culture after the incubation when compared to the control group. In addition we verified the plasmatic concentration of arctiin after intraperitoneal administration in mice by chromatographic analysis. The analysis of pure arctiin diluted in either methanol or mouse plasma showed a peak in the chromatogram at retention time of 5 minutes, absorbance was measured at 254 nm. Animal plasma sample collected one hour after treatment with arctiin was analyzed under same experimental conditions and revealed a similar peak, confirming the availability of arctiin in the plasma following administration. The in vivo tests were performed in Swiss female mice, those were intraperitoneally injected with two dosages of arctiin (50 mg/kg) - the first administered 20 days after infection and the second two weeks later. The follow parameters were analyzed: liver weight, white blood cell count, parasitic load and oogram. We did not find any significant change in those parameters comparing infected control groups treated either with praziquantel (200 mg / kg) or dimethyl sulfoxide (0.5 %) to the group treated with arctiin. The most promising result was the reduction around 20% of the average area of the granuloma in the arctiin group compared with the infected control. More studies are needed to verify possible mechanisms of action of this molecule in inflammatory components that play a role in granuloma formation.

Estudaram-se os efeitos da influência do Reiki na evolução do granuloma induzido experimentalmente pela inoculação do BCG no coxim plantar de hamsters, assim como os efeitos da mesma terapia em camundongos portadores do tumor ascítico de Ehrlich in vivo e in vitro. No modelo de inflamação granulomatosa crônica, utilizou-se 40 hamsters machos, os quais após serem inoculados com BCG no dia 0 no coxim da pata posterior direita, foram separados em dois grupos contendo 20 animais em cada. Um grupo denominado controle o qual não recebeu nenhum tratamento, e um grupo denominado Reiki, o qual recebeu reiki por 15 minutos, diariamente, a uma distância de 30 cm. Imediatamente antes da inoculação foi realizada a medida do diâmetro da pata a ser inoculada, sendo que as medições continuaram até o momento do sacrifício dos animais, sendo realizadas em dias alternados. No modelo de tumor ascítico de Ehrlich, foram utilzadas 26 camundongos fêmeas, as quais foram injetadas com células de Tumor de Ehrlich pela via intraperitoneal, sendo este considerado dia 0 do experimento. Em seguida estes camundongos foram separados em 3 grupos: controle (n=8), reiki A (n=9) e reiki B (n=9). Grupo controle não recebeu nenhum tipo de tratamento, grupo Reiki A recebeu reiki por 10 minutos, diariamente, à uma distância de 30 cm e grupo Reiki B recebeu manipulação e uma técnica diferente de Reiki. Os animais então foram observados diariamente até o dia de óbito. Em relação ao edema de pata, foi observada diminuição deste nos hamsters inoculados com BCG e tratados com Reiki. Com respeito à avaliação da taxa de sobrevida em camundongos com tumor ascítico de Ehrlich, observou-se maior taxa de sobrevida nos camundongos inoculados pertencentes ao grupo Reiki A.
We studied the effects of the influence of Reiki in the evolution of the experimental induced granuloma by the inoculation of BCG in the footpad of hamsters, and the effects of the same therapy in mice with Ehrlich ascitic tumor in vivo and in vitro. In the chronic granulomatous inflammation model, it was used, 40 male hamsters, which, after been inoculated with BCG in day 0, it was separated in two groups with 20 animals per group: control and reiki. The control group received no treatment, and reiki group, which was treated with Reiki by 15 minutes, daily, from 30 cm of the box. Immediately before the inoculation, the footpad diameter was measured, and after this measure was made in every other day, until complete 54 days. In the Ehrlich ascitic tumor model, it was used, 26 female mice inoculated with Ehrlich tumor cells, by intraperitoneal via, in the day 0. After that, the mice were separated in three groups: control (n=8), reiki A (n=9) and reiki B (n=9). The control group received no treatment, reiki A group received Reiki treatment by 10 minutes, daily, from 30 cm of the box, and reiki B group, which was manipulated and received a different way of Reiki treatment. The mice were observed daily until death in order to analyze survival rate. As regards granuloma evaluation it was observed a reduced footpad edema in hamsters inoculated with BCG and treated with Reiki. In relation to survival rate assay, it was observed an increased lifespan of those mice of the reiki A group.

Human granulosa cells play an important role in the follicle, providing the oocyte with nutrients and growth factors to ensure successful ovulation. Normal granulosa cell functioning is thus crucial to human fertility. By studying their transcriptome, the mechanisms underpinning follicle development and infertility will be better understood. In this study, granulosa cells were retrieved at the time of oocyte removal for in vitro fertilization (IVF). Cells were purified through a combination of a Percoll gradient to remove red blood cells and positive selection of granulosa cell aggregations. On average, contamination by white blood cells was 2% as measured by FACS analysis using specific white blood cell markers. Histological and electron microscopy of granulosa cell aggregations did not detect evidence of resident ovarian white blood cells. This technique provided a good source of pure, healthy granulosa cells for RNA extraction and subsequent gene expression studies. The construction of a human granulosa cell SAGE library derived 1689 SAGEtags and 1289 discrete mRNA transcripts. SAGEtags for a number of well recognized granulosa cell genes (FSH receptor, follistatin, connexin 43) were found in addition to hormone receptors, multiple kinases, structural genes, apoptosis related genes and secreted proteins. A variety of other SAGE libraries were downloaded from SAGEmap (two ovary epithelium, ovary, white blood cell, brain, cerebellum, heart, liver, lung, kidney, pancreas and universal human reference) and compared to the human granulosa cell library. This was based on two measures: a gene specificity score (GSS) and a tissue specificity score (TSS). Three SAGEtags were identified with high levels of expression in granulosa cells, but no or low expression in the other libraries. These were retinol binding protein 1 (RBP 1), scavenger receptor class B member 1 (SCARB 1) and hydroxysteroid (11-beta) dehydrogenase 1 (11-β-HSD). Library comparisons were validated by real time RT-PCR. The TSS score revealed granulosa cells were most similar to the universal reference library and least similar to liver. Granulosa cell samples were collected from woman undergoing IVF for a range of infertility disorders. These included polycystic ovary syndrome (PCOS), tubal disease, endometriosis and idiopathic (unknown). Gene expression was compared between these groups using real time RT-PCR. Candidate genes included RBP 1, 11-β-HSD, SCARB 1, FSH receptor, follistatin, decidual protein induced by progesterone, and progesterone membrane receptor component 1 and 2. Granulosa cell gene expression was significantly different (p<0.05) from human white blood cells. No differences were found in gene expression levels between the infertility disorders. Analysis of each patient, however, revealed individuals with marked over-expression of selected genes. The technique of Generation of Longer cDNA fragments for Gene Identification (GLGI) was used to investigate seven SAGEtags that did match known genes or ESTs. Although no novel genes were characterized, a further 14 granulosa cell transcripts were identified by this technique. This thesis used an integrated approach to the study of human granulosa cell gene expression. This has involved development of a purification method, the use of a high-throughput technique (SAGE), bioinformatics tools to identify candidates genes, real time RT-PCR to investigate gene expression of particular genes in infertility disorders and finally a technique with the potential to characterize unknown SAGEtags (GLGI). This systematic approach has advanced the understanding of gene expression in human granulosa cells and identified avenues for future research into folliculogenesis and human infertility.

As bactérias desmpenham um importante papel na etiopatogenia das doenças pulpares e periapicais. Com o objetivo de analisar a distribuição das bactérias nas estruturas mineralizadas de dentes portadores de necrose pulpar e nos granulomas apicais, utilizaram-se 32 raízes dentárias com lesões periapicais, firmemente aderidas a seus ápices e 16 lesões isoladas, em cortes obtidos para fins de diagnóstico microscópico com laudos histopatológicos conclusivos de granulomas apicais. Os ápices foram analisados pela microscopia óptica, empregando-se as técnicas de coloração pela hematoxilina-eosina de Harris e de Brown e Brenn. Os resultados evidenciaram alta freqüência de bactérias Gram-positivas e Gram-negativas no lume do canal principal e das ramificações constituintes do delta apical; e também, em menor proporção, nos túbulos dentinários, nos cementoplastos, na superfície radicular externa e nos granulomas apicais. A partir dos resultados obtidos, consideramdo-se as limitações inerentes à metodologia empregada, pôde-se verificar: 1 - uma elevada freqüência de bactérias no lume do canal do terço apical radicular, com predominância de cocos e bacilos Gram-positivos e Gram-negativos, distribuídos isoladamente ou em colônias, por vezes aderidos à parede do canal; 2 - a presença de bactérias nos túbulos dentinários do terço apical radicular, esendendo-se do terço pulpar circunferencial ao terço superficial circunferencial da dentina, nas formas de cocos e bacilos Gram-positivos e Gram-negativos; 3 - a presença de bactérias Gram-positivas e Gram-negativas nos granulomas apicais, com predomínio de cocos e bacilos; 4 - a presença de bactérias Gram-positivas e Gram-negativas nos granulomas apicais, com predomínio de cocos e bacilos, distribuídos em colônias ou isoladamente, nos espaços extracelulares e/ou no interior de células macrofágicas. Assim, pôde-se concluir: nos dentes com necrose pupar e granulomas apicais, as bactérias envolvem todo o sistema de canal radicular, incluindo-se os túbulos dentinários, e freqüentemente os tecidos da região periapical. Mais uma vez, fundamenta-se a necessidade do preparo biomecânico auxiliado por substâncias químicas antibacterianas, emprego de curativos no interior do canal, além do selamento adequado do sistema de canal radicular, em tratamentos endodônticos, além do selamento adequado do sistema de canal radicular, em tratamentos endodônticos de dentes com necrose pulpar e lesão periapical crônica.
Bacteria play a major role in the pathogeneses of pulpal and periapical diseases. The purpose of this study was to analyze bacterial arrangement in the hard structure of non-vital teeth with apical granulomas. We used 32 tooth roots with periapical lesions firmly adhered to them. For this study we also used 16 lesions, in slices obtained with the purpose of diagnosis, with compatible diagnosis of apical granulomas. The specimens were analyzed through optical microscopy using hematoxilin-eosin, and Brown and Brenn stain techniques. The results of this study showed a great number of Gram positive and Gram negative bacteria at the lumen of the main root and accessory canals; less amount in the dentinal tubules, in lacunae of the cellular cementum, on the root surface, and apical granulomas. Considering the metodology applied, our findings showed: 1 - A high frequency of bacteria in apical canal roots, where predominated Gram positive and Gram negative cocci and rods, distributed isolated or in bacterial colonies, sometimes attached in the wall canal root; 2 - In roots cutted in a transversal way we found the presence of Gram positive and Gram negative cocci and rods in dentinal tubules of the root third apical expanding to the third pulpal through the superficial layer dentin. 3 - The presence of Gram positive and Gram negative in the external surface of apical dentin root distributed isolated or plaques firmly adhered, where cocci and rods predominated. 4 - The presence of Gram positive and Gram negative, mainly cocci and rods, arranged in colonies or isolated in the external cellular spaces or within macrophage cells. Hence, we concluded that in teeth with necrotic pulps and apical granulomas the bacteria wrap up all root canal system includind dentinal tubules and the periapical region. This study enhanced once more the need of mechanical cleaning aided by chemical irrigation, antibacterial intracanal dressings and adequate filling of the root canal system in non-vital teeth with chronic periapical lesions.

A schistosomose é uma doença parasitária que afecta cerca de 200 milhões de pessoas, com alta prevalência nos trópicos e que origina um grave problema de saúde pública. Ao longo da infecção, o sistema imunitário tenta de várias formas combater a presença do parasita. Inicialmente ocorre uma resposta imune mediada por células do tipo Th1, com o progresso da infecção, a resposta é substituída por uma resposta do tipo Th2 induzida durante a formação de granulomas. Este surge como resposta à presença de produtos tóxicos libertados pelos ovos do parasita retido nos tecidos. O fígado é o principal alvo do depósito de ovos, sofrendo alterações fisiopatológicas, e histológicas. O Mus musculus tem sido muito utilizados na infecção experimental por Schistosoma mansoni, para melhor se conhecer o papel da resposta imunitária na formação de granulomas hepáticos. No decorrer da infecção o granuloma sofre alterações desencadeadas pelas citocinas que o sistema imunitário produz. Estas alterações dividem-se em cinco fases: reacção inicial, exsudativa, exsudativa-produtiva, produtiva e involutiva granuloma. O presente trabalho, estudou as alterações sofridas pelo granuloma hepático (quantidade, dimensão e fase do granuloma), em três diferentes períodos de infecção (55, 90 e 125 dias) no modelo animal Mus musculus infectado com Schistosoma. mansoni, estirpe SmBh distribuídos por três grupos experimentais com diferente número de cercárias (50, 80, e 100). Verificou-se que ao longo da infecção a quantidade de granulomas aumenta, as dimensões têm uma tendência inicial para aumentar mas a partir dos 90 dias após a exposição sofrem uma diminuição. No grupo experimental com maior intensidade de infecção inicial a diminuição deu-se mais cedo. Em relação às fases de desenvolvimento do granuloma este sofre alterações ao longo de toda a infecção. Assim, aos 55 dias predomina a fase exsudativa, aos 90 todos os grupos apresentam maior percentagem de granulomas na fase produtiva e por fim aos 125 dias prevalece a fase involutiva. Todos estes resultados sugerem que a caracterização do granuloma nas diferentes fases de infecção pode depender do número de cercárias da exposição.
Schistosomiasis is a parasitic disease that affects about 200 million people worldwide, with high prevalence in the tropics and causes a serious public health problem. Throughout infection, the immune system tries various ways to combat parasites. Initially there is an immune response mediated by Th1 cells, with the progress of the infection, the response is replaced by a Th2 type response induced during the formation of granulomas. This is a response to the presence of toxic products released by the parasite eggs retained in tissues. The liver is the main target for the deposition of eggs and suffers pathophysiological and histological changes. Mus musculus has been widely used for experimental infection by Schistosoma mansoni, to better understand the role of the immune response in the formation of hepatic granulomas. Throughout the infection, the granuloma undergoes changes triggered by the type of cytokines that the immune system produces. These changes are divided into five stages: initial reaction, exudative, exudative-productive, productive and involutional granuloma. The present study investigated the changes undergone by hepatic granuloma (quantity, size and stage of granuloma) in three different stages of infection (55, 90 and 125 days) in a Mus musculus animal model infected with S. mansoni strain SmBh divided into three groups with different number of cercariae (50, 80 and 100 cercariae). It was found that in the course of the infection the amount of granulomas increases. Their dimensions have an initial tendency to increase but after 90 days of infection they start to decrease. In the experimental group with a higher intensity of initial infection the decline took place earlier. In relation to the phases that the granuloma undergoes throughout the infection, at 55 days dominates the exudative phase, at 90 days all groups have a higher percentage of granulomas in the production phase and finally to 125 days of infection prevails involution phase. All these results suggest that the characterization of the granuloma in different stages of infection may be dependent on the intensity of initial infection.