Orientador: Paulo Câmara Marques Pereira / Resumo: A Doença de Chagas, causada pelo Trypanosoma cruzi (T. cruzi), é uma patologia infecciosa crônica que apresenta grande incidência no Brasil e, nos últimos anos, passou a ser endêmica em centros urbanos. É acompanhada por alterações no sistema imune com variações nos níveis séricos das citocinas pró-inflamatórias. O tratamento disponível no Brasil é o benzonidazol, com baixa eficácia na fase crônica e efeitos colaterais importantes. Sabe-se que os adipocitos são reservatórios de T. cruzi, aspecto importante que é a relação com as citocinas pro-inflamatórias e com o aumento de T.cruzi nos adipocitos interferindo na evolução da doença. Sendo assim, o objetivo do presente estudo foi avaliar os efeitos da obesidade e das citocinas pro inflamatórias em camundongos balb/c machos infectados com T. cruzi tratados ou não com benzonidazol. Foram utilizados 120 animais, divididos em seis grupos, sendo 3 com dieta normal e 3 com dieta hipercalórica. Esses foram avaliados em cinco momentos: M1-1°dia, M2-3°dia, M3-5°dia, M4-7°dia e M5-8°dia. Para infecção utilizou a cepa aguda Y, com pico de parasitemia no sétimo dia (M4). A eutanásia foi realizada em cinco animais de cada grupo, em cada momento. Para determinação das citocinas utilizou o Kits de CBA –Cytometric Beads Array, através de citômetro de fluxo. Além disso, foi verificado o peso da gordura. O medicamento foi ofertado na dosagem de 100 mg/Kg/dia. A análise estatística foi realizada por medidas descritivas com o cálculo de média, de... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Chagas disease, caused by Trypanosoma cruzi (T. cruzi), is a chronic infectious disease that has a high incidence in Brazil and, in recent years, has become endemic in urban centers. It is accompanied by changes in the immune system with variations in serum levels of pro-inflammatory cytokines. The available treatment in Brazil is benzonidazole, with low efficacy in the chronic phase and important side effects. It is known that the adipocytes are reservoirs of T. cruzi, an important aspect that is the relation with the pro-inflammatory cytokines and with the increase of T. cruzi in the adipocytes interfering in the evolution of the disease. Therefore, the objective of the present study was to evaluate the effects of obesity and proinflammatory cytokines in T. cruzi infected or non-treated male balb / c mice with benzonidazole. A total of 120 animals were used, divided into six groups: 3 with normal diet and 3 with hypercaloric diet. These were evaluated in five moments: M1-1 day, M2-3 day, M3-5 day, M4-7 day and M5-8 day. For infection, the acute Y strain was used, with a peak of parasitemia on the seventh day (M4). Euthanasia was performed in five animals from each group, at each moment. In order to determine the cytokines, the CBA-Cytometric Beads Array Kits were used by flow cytometer. In addition, the fat weight was checked. The drug was offered in the dosage of 100 mg / kg / day. Statistical analysis was performed by descriptive measures with mean, standard deviation, mi... (Complete abstract click electronic access below) / Doutor

O aparelho ortodôntico promove alterações microbiológicas na cavidade bucal, em função da variedade de materiais sólidos e elásticos que possuem, os quais funcionam como áreas de retenção, levando a um acúmulo de biofilme e predispondo o hospedeiro à cárie dental e à doença periodontal. Os objetivos do presente estudo, in vivo, foram: 1) Avaliar, as alterações no índice de placa (PI), índice gengival (GI), índice de sangramento gengival (GBI) e no volume do fluido crevicular, em pacientes com 3 diferentes desenhos de bráquetes metálicos (autoligados e convencionais), a fim de verificar se o desenho dos bráquetes interfere no acúmulo de placa e na saúde gengival; 2) Estudar as alterações nos níveis de diferentes micro-organismos envolvidos direta ou indiretamente na cárie dental, na saliva e in situ, pré e pós-colagem de bráquetes convencionais e autoligados; 3) Avaliar os níveis salivares e in situ, de espécies de micro-organismos dos complexos roxo, verde, laranja e vermelho e de outras espécies, pré e pós-colagem de bráquetes convencionais e autoligados, por meio de sondas de DNA e 4) Quantificar citocinas pró-inflamatórias no fluido crevicular, antes e após a colagem dos diferentes bráquetes. A amostra foi constituída de 20 pacientes, com idade entre 11 e 15 anos (Média: 13,3 anos), que receberam bráquetes metálicos convencionais GeminiTM e dois diferentes tipos de bráquetes autoligados: In-Ovation®R e SmartClipTM em incisivos e caninos superiores. Os índices, o volume do fluido e as amostra de saliva (S0) foram obtidos antes da instalação dos aparelhos (T0) e após 30(T1) e 60(T2) dias. Um bráquete de cada tipo foi removido 30 e 60 dias após a colagem. Os dados foram submetidos à análise estatística utilizando-se os testes não-paramétricos de Friedman, Mann-Whitney e Wilcoxon e coeficiente de correlação não-paramétrico de Spearman. A análise da concentração de citocinas no fluido crevicular foi realizada segundo modelo de análise de variância misto, seguida pelo teste de Tukey, com nível de significância de 5%. Não houve correlação entre o grau de apinhamento, overjet e overbite com os escores do índice de placa (PI), índice gengival, índice de sangramento gengival e com o volume do fluido crevicular no tempo inicial T0 (p>0,05). Verificou-se a diferença significativa nos escores de PI e volume do fluido crevicular somente nos dentes que receberam bráquetes autoligados SmartClipTM, entre os tempos T0-T2(p<0,05). Identificou-se a presença de 21 das 22 espécies bacterianas e 4 das 5 espécies fúngicas, na saliva, antes da colagem dos aparelhos ortodônticos. In situ, no bráquete autoligado SmartClipTM observou-se maiores níveis de espécies do complexo vermelho 60 dias após a colagem. Para os bráquetes convencionais GeminiTM verificou-se diminuição nos níveis de espécies do complexo roxo. Para o bráquete autoligado In-Ovation®R identificou-se diferença significativa para C. rectus 30 dias após a colagem e para S. mutans 60 dias após a colagem. Quando comparou-se os níveis salivares e os in situ, verificou-se maiores níveis de espécies nos bráquetes, in situ, sendo que os autoligados apresentaram os maiores valores. Aumento significante foi verificado para os bráquetes autoligados In-Ovation®R, para S. mutans e L. casei. Após 60 dias da colagem do aparelho ortodôntico observou-se um aumento nos níveis de TNF-&alpha; (p<0.005), em todos os tipos de bráquetes. Considerando-se as condições específicas desse estudo, pode-se concluir que o desenho do bráquete modulou o índice de placa, o volume do fluido crevicular, e os níveis bacterianos do complexo vermelho, sendo o bráquete autoligado SmartClipTM o que apresentou o pior desempenho, nesse aspecto. O bráquete autoligado In-Ovation®R apresentou maiores níveis bacterianos relacionados à cárie dental e espécies do complexo laranja. A concentração de citocinas no fluido crevicular não foi modulada pelo desenho do bráquete, ocorrendo um aumento na concentração de TNF-&alpha; nos três tipos de bráquetes (convencional e autoligados) 60 dias após a colagem. A análise global dos resultados obtidos nos diferentes parâmetros analisados no presente estudo permitiu evidenciar, em geral, melhores resultados clínicos com o bráquete metálico convencional GeminiTM. / The orthodontic appliance promoting several changes in oral cavity according to the variety of solid and elastic materials used in the treatment, leading to the biofilm development. The aim of this study, in vivo, was: 1) Evaluate the periodontal index: Plaque Index (PI), Gingival Index (GI), Gingival Bleeding Index (GBI)and gingival crevicular fluid volume. 2) The levels of cariogenic and periodontopatogenical bacteria in saliva and in two different metallic brackets: self-ligating and conventional, quantify the levels before and 30 and 60 days after bonding orthodontic appliance. 3) The levels of red, orange green and purple complex and others species, in situ and in the saliva. 4) The pro-inflammatory cytokines levels in the gingival crevicular fluid. The sample consisted of 20 patients, aged between 11 and 15 years (mean 13.3 years) who received conventional metal bracket GeminiTM and two different types of self-ligating brackets: In-Ovation®R and SmartClipTM in maxillary incisors and canines. The rates, the volume of the fluid and saliva sample (S0) were obtained before the device (T0), 30 (T1) and 60 (T2) after bonding. A type of each bracket was removed 30 to 60 days after bonding. Data were analyzed statistically using the nonparametric Friedman test, Mann-Whitney and Wilcoxon coefficient and nonparametric Spearman correlation with a significance level of 5 %. The cytokine levels was analyzed by the mixed models, follow to the Tukey test. There was no correlation among the scores of crowding, overjet and overbite with the scores of plaque index (PI), gingival index (GI), gingival bleeding index (GBI) and volume of crevicular fluid at the initial time T0 (p>0.05). There was a significantly difference in PI and crevicular fluid volume, when compared each bracket separately, only the teeth that received self-ligating bracket SmartClipTM, between times T0-T2 (p<0.05). We identified, in the saliva, the presence of 21, bacterial species, of the 22 analyzed; 4 of the 5 fungal species, before bonding the orthodontic appliance. In situ, to the SmartClipTM the highest levels were observed to the red complex 60 days after bonding. To the GeminiTM a decreased to the purple complex was observed when compared 30 and 60 days after bonding. The In-Ovation®R showed a significantly difference to C. rectus, 30 days after bonding. S. mutans, 60 days after bonding. 30 days after bonding the microbial levels is highest in the saliva than in situ, the self-ligating brackets present the highest levels than conventional brackets. 60 days after bonding the TNF-&alpha; levels was increased (p<0,05). The microbial levels was highest in the self-ligating brackets than the conventional brackets. The bracket design seems influenced the plaque index, the volume of the crevicular fluid, the microbial adhesion. The levels in situ are highest than in saliva. The TNF-&alpha; levels were increased 60 days after bonding. The bracket design change the PI, gingival crevicular volume and the red complex species, the SmartClipTM showed the worst results in this point. The In-Ovation®R presented the highest levels of the species to the orange complex and cariogenic species. The design didn\'t modulated the cytokine levels, th TNF&alpha;, increased for all brackets. The global analyses of the results allows us to conclude that the GeminiTM conventional bracket showed the best results when compared to the others self-ligating.

O aparelho ortodôntico promove alterações microbiológicas na cavidade bucal, em função da variedade de materiais sólidos e elásticos que possuem, os quais funcionam como áreas de retenção, levando a um acúmulo de biofilme e predispondo o hospedeiro à cárie dental e à doença periodontal. Os objetivos do presente estudo, in vivo, foram: 1) Avaliar, as alterações no índice de placa (PI), índice gengival (GI), índice de sangramento gengival (GBI) e no volume do fluido crevicular, em pacientes com 3 diferentes desenhos de bráquetes metálicos (autoligados e convencionais), a fim de verificar se o desenho dos bráquetes interfere no acúmulo de placa e na saúde gengival; 2) Estudar as alterações nos níveis de diferentes micro-organismos envolvidos direta ou indiretamente na cárie dental, na saliva e in situ, pré e pós-colagem de bráquetes convencionais e autoligados; 3) Avaliar os níveis salivares e in situ, de espécies de micro-organismos dos complexos roxo, verde, laranja e vermelho e de outras espécies, pré e pós-colagem de bráquetes convencionais e autoligados, por meio de sondas de DNA e 4) Quantificar citocinas pró-inflamatórias no fluido crevicular, antes e após a colagem dos diferentes bráquetes. A amostra foi constituída de 20 pacientes, com idade entre 11 e 15 anos (Média: 13,3 anos), que receberam bráquetes metálicos convencionais GeminiTM e dois diferentes tipos de bráquetes autoligados: In-Ovation®R e SmartClipTM em incisivos e caninos superiores. Os índices, o volume do fluido e as amostra de saliva (S0) foram obtidos antes da instalação dos aparelhos (T0) e após 30(T1) e 60(T2) dias. Um bráquete de cada tipo foi removido 30 e 60 dias após a colagem. Os dados foram submetidos à análise estatística utilizando-se os testes não-paramétricos de Friedman, Mann-Whitney e Wilcoxon e coeficiente de correlação não-paramétrico de Spearman. A análise da concentração de citocinas no fluido crevicular foi realizada segundo modelo de análise de variância misto, seguida pelo teste de Tukey, com nível de significância de 5%. Não houve correlação entre o grau de apinhamento, overjet e overbite com os escores do índice de placa (PI), índice gengival, índice de sangramento gengival e com o volume do fluido crevicular no tempo inicial T0 (p>0,05). Verificou-se a diferença significativa nos escores de PI e volume do fluido crevicular somente nos dentes que receberam bráquetes autoligados SmartClipTM, entre os tempos T0-T2(p<0,05). Identificou-se a presença de 21 das 22 espécies bacterianas e 4 das 5 espécies fúngicas, na saliva, antes da colagem dos aparelhos ortodônticos. In situ, no bráquete autoligado SmartClipTM observou-se maiores níveis de espécies do complexo vermelho 60 dias após a colagem. Para os bráquetes convencionais GeminiTM verificou-se diminuição nos níveis de espécies do complexo roxo. Para o bráquete autoligado In-Ovation®R identificou-se diferença significativa para C. rectus 30 dias após a colagem e para S. mutans 60 dias após a colagem. Quando comparou-se os níveis salivares e os in situ, verificou-se maiores níveis de espécies nos bráquetes, in situ, sendo que os autoligados apresentaram os maiores valores. Aumento significante foi verificado para os bráquetes autoligados In-Ovation®R, para S. mutans e L. casei. Após 60 dias da colagem do aparelho ortodôntico observou-se um aumento nos níveis de TNF-&alpha; (p<0.005), em todos os tipos de bráquetes. Considerando-se as condições específicas desse estudo, pode-se concluir que o desenho do bráquete modulou o índice de placa, o volume do fluido crevicular, e os níveis bacterianos do complexo vermelho, sendo o bráquete autoligado SmartClipTM o que apresentou o pior desempenho, nesse aspecto. O bráquete autoligado In-Ovation®R apresentou maiores níveis bacterianos relacionados à cárie dental e espécies do complexo laranja. A concentração de citocinas no fluido crevicular não foi modulada pelo desenho do bráquete, ocorrendo um aumento na concentração de TNF-&alpha; nos três tipos de bráquetes (convencional e autoligados) 60 dias após a colagem. A análise global dos resultados obtidos nos diferentes parâmetros analisados no presente estudo permitiu evidenciar, em geral, melhores resultados clínicos com o bráquete metálico convencional GeminiTM. / The orthodontic appliance promoting several changes in oral cavity according to the variety of solid and elastic materials used in the treatment, leading to the biofilm development. The aim of this study, in vivo, was: 1) Evaluate the periodontal index: Plaque Index (PI), Gingival Index (GI), Gingival Bleeding Index (GBI)and gingival crevicular fluid volume. 2) The levels of cariogenic and periodontopatogenical bacteria in saliva and in two different metallic brackets: self-ligating and conventional, quantify the levels before and 30 and 60 days after bonding orthodontic appliance. 3) The levels of red, orange green and purple complex and others species, in situ and in the saliva. 4) The pro-inflammatory cytokines levels in the gingival crevicular fluid. The sample consisted of 20 patients, aged between 11 and 15 years (mean 13.3 years) who received conventional metal bracket GeminiTM and two different types of self-ligating brackets: In-Ovation®R and SmartClipTM in maxillary incisors and canines. The rates, the volume of the fluid and saliva sample (S0) were obtained before the device (T0), 30 (T1) and 60 (T2) after bonding. A type of each bracket was removed 30 to 60 days after bonding. Data were analyzed statistically using the nonparametric Friedman test, Mann-Whitney and Wilcoxon coefficient and nonparametric Spearman correlation with a significance level of 5 %. The cytokine levels was analyzed by the mixed models, follow to the Tukey test. There was no correlation among the scores of crowding, overjet and overbite with the scores of plaque index (PI), gingival index (GI), gingival bleeding index (GBI) and volume of crevicular fluid at the initial time T0 (p>0.05). There was a significantly difference in PI and crevicular fluid volume, when compared each bracket separately, only the teeth that received self-ligating bracket SmartClipTM, between times T0-T2 (p<0.05). We identified, in the saliva, the presence of 21, bacterial species, of the 22 analyzed; 4 of the 5 fungal species, before bonding the orthodontic appliance. In situ, to the SmartClipTM the highest levels were observed to the red complex 60 days after bonding. To the GeminiTM a decreased to the purple complex was observed when compared 30 and 60 days after bonding. The In-Ovation®R showed a significantly difference to C. rectus, 30 days after bonding. S. mutans, 60 days after bonding. 30 days after bonding the microbial levels is highest in the saliva than in situ, the self-ligating brackets present the highest levels than conventional brackets. 60 days after bonding the TNF-&alpha; levels was increased (p<0,05). The microbial levels was highest in the self-ligating brackets than the conventional brackets. The bracket design seems influenced the plaque index, the volume of the crevicular fluid, the microbial adhesion. The levels in situ are highest than in saliva. The TNF-&alpha; levels were increased 60 days after bonding. The bracket design change the PI, gingival crevicular volume and the red complex species, the SmartClipTM showed the worst results in this point. The In-Ovation®R presented the highest levels of the species to the orange complex and cariogenic species. The design didn\'t modulated the cytokine levels, th TNF&alpha;, increased for all brackets. The global analyses of the results allows us to conclude that the GeminiTM conventional bracket showed the best results when compared to the others self-ligating.

A doença periodontal é resultado da inflamação das estruturas periodontais em resposta ao biofilme dentário presente na superfície dental e sulco gengival. Sua etiopatogenia é multifatorial e complexa. Na periodontite crônica há bacteremia freqüente durante rotinas diárias como escovação dentária e mastigação e acredita-se que esse estímulo constante ao sistema imunológico possa causar repercussões sistêmicas nos pacientes, como arterioescleroses. Uma das formas atuais de se mensurar essa alteração é pela presença de mediadores inflamatórios séricos. Sendo as proteínas de fase aguda, como as citocinas, as mais avaliadas. Assim como os trabalhos encontrados para a espécie humana, propôsse a mensuração sérica das interleucinas IL-1, IL-6, IL-10 e TNF-&alpha; em cães com doença periodontal crônica. Como grupo controle, utilizaram-se os mesmos pacientes após tratamento periodontal, mensurando as mesmas interleucinas depois de 21 dias do tratamento. Como resultado todas as interleucinas sofreram alteração, porém somente a IL-6 teve redução estatisticamente significativa após o tratamento periodontal. Todavia atenta-se para os elevados valores encontrados em alguns pacientes, o que pode demonstrar importante alteração sistêmica. / Periodontal disease is the result of inflammation of the periodontal structures in response to the biofilm present on the tooth surface and the gingival sulcus. Its pathogenesis is multifactorial and complex. Bacteremia in chronic periodontitis is constant during daily routines, such as toothbrushing and chewing, and it is believed that this constant stimulus to the immune system can cause systemic effects in patients, such as arterioescleroses. One of the current ways of measuring this change is the presence of serum inflammatory mediators. The acute phase proteins, such as cytokines, are the most valued. Like the works found for the human species, the measurement of serum interleukins IL-1, IL-6, IL- 10 and TNF-&alpha; in dogs with chronic periodontal disease was proposed. As controls, we used the same patients after periodontal treatment, measuring the same interleukins after 21 days of treatment. As a result, all interleukins have been changed, but only IL-6 showed statistically significant decrease after periodontal treatment. However, the high values found in some patients, which may prove important systemic change, must be highlighted.

A Asma vem apresentando taxas de prevalência crescentes em todo o mundo. Os objetivos deste trabalho são avaliar a presença de elementos celulares e humorais indicativos em sangue e colostro de mães asmáticas e saudáveis e no sangue de cordão umbilical de seus recém-nascidos (RN). Observamos menor produção de IgG pelas mães asmáticas. Células dendríticas de mães asmáticas possuem maior expressão de CD80 e CD86. Mães asmáticas possuem mais células de memória central. Linfócitos T CD4+ de mães asmáticas produzem níveis maiores IFN-g. Células CD3+ e CD4+ de mães asmáticas produziram mais IL-13. Mães saudáveis produziram maiores quantidades de IL-10. Concluímos que mães asmáticas possuem menores níveis de IgG e IgM, o que parece aumentar dos níveis de IgE, mães asmáticas possuem mais células de memória central, linfócitos T CD4+ produzem maiores quantidades de citocinas como IL-13 e IFN-g, células dendríticas de mães asmáticas possuem maior expressão das moléculas co-estimulatórias, assim como seus RNs possuem maior expressão de CD80. Células de mães asmáticas produzem níveis menores de IL-10, o colostro de mães atópicas não possui diferenças entre os parâmetros aqui estudados. O aleitamento materno deve ser indicado para mães asmáticas e seus filhos. / Asthma has been presenting higher prevalence rates worldwide. The objective of this work is to evaluate the presence of cellular elements and humoral indicative in blood and colostrums of healthy and asthmatic mothers and in cord umbilical blood of their newborn (NB). Lower production of IgG from asthmatic mothers was observed. Dendritic cells of asthmatic mothers have higher CD80 and CD86 expression. Asthmatic mothers have more central memory cells. TCD4+ lymphocyte of asthmatic mothers produce higher levels of IFN-g. CD3+ and CD4+ cells of asthmatic mothers produce higher quantity of IL-13. Healthy mothers produce higher quantity of IL-10. We conclude that asthmatic mothers have lower levels of IgG and IgM, and this seems to raise the IgE levels, asthmatic mothers have more central memory cells, CD4+ T lymphocyte and produce higher quantities of cytokines such as Il-13 and IFN-g. Dendritic cells of asthmatic mothers have higher expression of costimulatory molecules, as well as their NBs have higher expression CD80. Cells of asthmatic mothers produce lower levels of IL-10, the colostrums of atopic mothers does not have differences in the parameters here studied. The maternal breastfeeding must be indicated for asthmatic mothers and their of spring.

Over one third of the human population is currently infected by one or more species of parasitic helminth, but the immune responses elicited by these infections remain poorly defined. Studies in helminth-exposed human populations and laboratory models suggest that helminth infection elicits a range of different effector cell types and that protective immunity and resistance to immune-mediated pathology depends on the balance between these responses. The aim of this thesis was to investigate how cytokines, the molecular mediators of the immune system, can be used to characterise human immune phenotype during natural and experimental helminth infection. Cytokines associated with innate inflammatory (TNFα, IL-6 and IL-9), Thl (IFNγ, IL-2 and IL-12p70), Th2 (IL-4, IL-5 and IL-13), Th17 (IL-17A, IL-21 and IL-23) and regulatory (IL-10 and TGFβ)immune phenotypes were analysed to provide the most comprehensive analysis of cytokine responses in human helminth infection conducted to-date. Using a multivariate statistical approach cytokines were analysed as combined immune profiles to reflect their complex interactions in vivo. In the first part of the study venous blood samples collected from a cross-sectional cohort of 284 Zimbabweans (age range: 3 -86 years) endemically-exposed to Schistosoma haematobium were cultured with antigens from different stages of the parasite's life-cycle(cercariae, adult worms and eggs) and the anti-schistosome vaccine candidate antigen glutathionine-S-transferase (GST). Cytokines responses were quantified in culture supernatants via enzyme-linked immunosorbent assay (ELISA). These assays were repeated 6 weeks after clearance of infection by anti-helminthic treatment. Parasitological and demographic characterisation of the cohort before, 6 weeks, 6 and 18 months after treatment allowed cytokine responses to be related to epidemiological patterns of infection before treatment and the risk of re-infection after treatment. The main findings of this study were:Cytokine responses to the antigens of S. haematobium cercariae are more proinflammatory than those elicited by adult worms and eggs prior to treatment, reflecting the distinct proteomes and exposure patterns of the 3 life-cycle stages Young children (5-10 years old) have a more regulatory and Th17-polarised cytokine response to S. haematobium antigens than older children and adults. These responses are significantly associated with schistosome infection intensity and may contribute to the development of resistance to schistosomiasis with age and exposure to infection Anti-helminthic treatment leads to a shift in S. haematobium cercariae, egg and GST specific cytokine responses towards a more pro-inflammatory phenotype The magnitude of change in S. haematobium-specific cytokine profiles after treatment is dependent on schistosome infection intensity at the time of treatment Individuals who remain un-infected up to 18 months after treatment to clear schistosome infection have a more pro-inflammatory and IL-21-polarised response to S. haematobium antigens 6 weeks after treatment than those who become re-infected, suggesting that post-treatment cytokine profiles promote resistance to re-infection. The second part of the study assayed systemic, parasite and allergen-specific cytokine responses in 45 adults with seasonally exacerbated allergy to grass pollen who were experimentally exposed to Trichuris suis. Cytokine responses in infected individuals were compared to those of 44 un-infected controls. This aspect of the study showed that: Exposure to T. suis promotes systemic and parasite-specific Th2 and regulatory cytokine responses, but does not alter cytokine responses to environmental allergens.

The biological basis of the clinical heterogeneity in Parkinson's Disease (PD) is unclear. It is likely to involve complex interactions between genetic and environmental factors and between a range of pathological processes, including protein homeostasis and immune system function. Microglial activation in the brain and peripheral innate immune changes are known to occur in PD. Recently genetic, animal and cellular studies have linked several innate immune related genes and proteins (e.g. HLADR, TREM2, TLR2, TLR4, caspase-1) to PD and provided evidence that they may have a role in PD pathogenesis. Alpha-synuclein is central to PD, with evidence from neuropathology, genetics and animal/cell models indicating that it plays a significant pathogenic role. There is developing evidence directly linking innate immune activity and alpha-synuclein pathology. For example, inflammation, particularly in response to microbial infection, is associated with increased alpha-synuclein accumulation in the periphery and activation of the innate immune inflammasome related caspase-1 leads to increased cleavage and aggregation of alpha-synuclein. Overall Hypothesis- "Parkinson's disease (PD) and its clinical heterogeneity are associated with systemic changes in innate immune and associated microbial factors and in alpha-synuclein". This was investigated from the perspective of an epidemiological study, a study of peripheral blood monocyte, innate immune/microbial markers and a cerebrospinal fluid (CSF) study in PD patients. *The epidemiological study, involved the longitudinal PICNICS cohort of 290 Idiopathic PD patients, and showed that the use of medication known to influence alpha-synuclein and immune function is associated with motor heterogeneity in PD. *The peripheral immune study involved 41 early PD patients and 41 age, gender and MAPT genotype matched paired controls, with the PD patients categorised into 2 groups based on the presence of previously identified clinical and genetic risk factors for the development of an early dementia (impaired semantic fluency, pentagon copying and MAPT H1/H1 haplotype). This study demonstrated that the phenotypic profile of peripheral monocytes and the level of serum alpha-synuclein and relevant innate immune and microbial markers do differ in early PD compared to controls and that there are differential changes in those patients at higher versus lower risk for early dementia. The systemic alpha-synuclein related changes appear to be present overall in PD patients compared to controls, while the more microbial/innate immune related changes appear to be more prominent in the dementia higher risk group. *The CSF study involved samples from 35 PD patients and has demonstrated evidence of relationships between neurodegeneration-linked CSF tau species and inflammatory cytokines, and between CSF alpha-synuclein and cognitive function, suggesting that these factors may be involved in PD heterogeneity within the central nervous system as well. Overall, these studies provide evidence that variations in alpha synuclein/ tau homeostasis and innate immune and microbial factors are related to PD and its clinical heterogeneity.

A sepse é caracterizada por uma produção excessiva de mediadores inflamatórios, acompanhada de taquicardia e hipotensão. Experimentalmente, a administração de endotoxina (Lipopolissacarídeo, LPS) em doses relativamente elevadas induz choque endotoxêmico, sendo um bom modelo de estudo da sepse. Diversos grupos têm demonstrado ações antiinflamatórias e antitumorais do citral, um composto do óleo essencial de Cymbopogon citratus. Nosso laboratório demonstrou ação antipirética do citral em modelo de febre induzida por LPS, acompanhada de redução nos níveis de citocinas plasmáticas e de prostaglandina E2 (PGE2) no plasma e área pré óptica do hipotálamo (POA), importante região termorregulatória. A hipótese testada neste trabalho foi a de que o citral atenua a hipotensão provocada pela endotoxina, além de amenizar as alterações termoregulatórias. Todos os procedimentos foram executados de acordo com os princípios éticos de experimentação animal, aprovados pelo comitê de ética local (CEUA 2015.1 1214 58-2). Foi realizado o implante de cânulas na artéria e veia femoral para registro da pressão arterial e administração de LPS (1,5 mg/kg) ou salina apirogênica 0,9% além do implante de datalogger na cavidade peritoneal de ratos Wistar, para registro da temperatura corporal. No dia do registro, 30 minutos antes da administração de LPS ou salina, os animais receberam citral (100 mg/kg) ou tween 80 a 1% (veículo) por via oral. Os parâmetros cardiovasculares e temperatura corporal foram registrados por 300 minutos após os respectivos tratamentos. Os valores de pressão arterial média (PAM) e frequência cardíaca (FC) foram coletados a cada 10 minutos após o tratamento e a temperatura corporal foi registrada pelo datalogger em intervalos de 5 minutos. Em outro protocolo foi realizado apenas o implante de cânula na veia femoral dos animais de todos os grupos para administração de LPS ou salina, coleta de sangue para dosagem de interleucina 6, PGE2, nitrito e nitrato e corticosterona e coleta do encéfalo para dosagem de PGE2 e PGD2. As diferenças estatísticas entre os grupos foram analisadas pelo teste ANOVA two-way seguido por pós teste de Newman-Keuls, com o nível de significância adotado de p < 0,05. A administração de LPS provocou queda na PAM eaumento na FC. Tais respostas não foram afetadas pela administração prévia de citral. O LPS também induziu febre e aumento nas concentrações plasmáticas de interleucina - 6 (IL-6), óxido nítrico (NO), PGE2 e corticosterona. Esses parâmetros não foram alterados pela pré- administração de Citral. No entanto, o citral provocou redução na produção de PGD2 naPOA, sem alterar a de PGE2 nesta região. Podemos concluir que o citral não previne as alterações nos parâmetros cardiovasculares no modelo de endotoxemia em ratos, porém reduz a produção de um mediador termorregulatório e inflamatório do sistema nervoso central (a PGD2), sem alterar a produção de outros mediadores inflamatórios a nível periférico (no plasma). Portanto, em um modelo mais agressivo de inflamação sistênica o citral não se mostrou suficiente para proteger o organismo das ações deletérias do LPS. / Sepsis is characterized by the overproduction of inflammatory mediators, accompanied by tachycardia and hypotension. Experimentally, administration of endotoxin (Lipopolysaccharide, LPS) in relatively high doses induces endotoxemic shock, a widely used model of sepsis in rats. Several groups have demonstrated anti-inflammatory and antitumor roles of citral, an essential oil compound of Cymbopogon citratus. Emilio-Silva et al. (2017) have shown an antipyretic role of citral in a model of LPS-induced fever, accompanied by a reduction of cytokines and prostaglandin E2 plasma levels and in the preoptic area of hypothalamus (POA), the hierarchically most important thermoregulatory region. We hypothesized that citral attenuates the LPS-induced hypotension, besides mitigating the thermoregulatory adjustments in rats. All procedures were performed in agreement with ethical guidelines for animal experimentation aproved by the local ethical committee (CEUA 2015.1 1214 58-2). Femoral artery and vein were implanted with cannulas for blood pressure recording and LPS (1.5 mg/kg) or 0.9% apyrogenic saline injection. In a second surgical procedure a datalogger was implanted into the peritoneal cavity for measurements of body temperature. All surgical procedures were performed under Ketamin/xilazin (100/10 mg/kg) anesthesia. One day after arterial catheterization, 30 minutes prior to LPS or saline administration, the animals received either citral (100 mg / kg) or 1% tween 80 (vehicle) oralstarly. The cardiovascular parameters and body temperature were recorded for 300 minutes after the respective treatments. Mean blood pressure (MAP) and heart rate (HR) were collected every 10 minutes after treatments and body temperature was recorded by the datalogger at 5 minute intervals. Blood samples were obtained in another set of rats for interleukin-6, PGE2, nitrite and nitrate and corticosterone analyses. The brain was removed for PGE2 and PGD2 analyses. Statistical differences between groups were analyzed by the two-way or one-way ANOVA test followed by Newman-Keuls post-test, with significance level adopted at p <0.05. As expected, LPS administration caused a decrease in MAP and an increase in HR, and these responses were not affected by citral. LPS also induced fever and increased plasma levels of interleukin - 6 (IL - 6), nitric oxide (NO), prostaglandin E 2 andcorticosterone. These parameters were also not altered by citral. On the other hand, citral caused a reduction in prostaglandin D2 concentration in the POA, but failed to alter PGE2 levels in this region. Our data are consistent with the notion that citral does not affect changes in cardiovascular and thermoregulatory parameters. Consistently, citral also caused no changes in both LPS-induced peripheral inflammatory mediators (in plasma) and in the POA, except PGD2. Therefore, in our model which mimetic a fairly critical situation, citral may not be sufficient to protect the organism from the deleterious actions of LPS. Financial support: FAPESP / CAPES.

Histatins, a group of proteins produced by human salivary glands, have a variety of innate immune functions including the ability to: kill oral microorganisms, neutralize toxins, inactivate protease/collagenase activities, inhibit co-aggregation of oral bacteria, and inhibit lipopolysaccharide mediated activities. Hemagglutinin B (HagB), a virulence factor of the periodontal pathogen Porphyromonas gingivalis, induces a robust cytokine and chemokine response in human myeloid dendritic cells. In this study, I hypothesize that histatin 5 can attenuate a HagB-induced chemokine response. Objectives: To characterize an expanded cytokine and chemokine response induced in human myeloid dendritic cells by HagB, and to determine if prior incubation of HagB with histatin 5 attenuates these responses. Methods: In my first experiment, 0.040 M HagB was mixed with dilutions of histatin 5 and histatin 8 (Sigma, 0.04 to 40.0 M), incubated at 37C for 30 minutes, and added to 2 x 104 human myeloid dendritic cells (Lonza, Walkersville, MD). At 24 hours, culture media was removed, and 6 cytokines and chemokines (pg/ml) were determined in cell-free supernatants (Millipore, Billerica, MA) using the Luminex 100 IS instrument (Luminex, Austin, TX). In my second experiment, 0.040 M HagB was mixed with 40.0 M histatin 5 only (e.g., 1:1000), incubated at 37C for 30 minutes, and added to 2 x 104 human myeloid dendritic cells. At 0, 1, 2, 4, 8, 16, and 24 hours post-inoculation, culture media was removed, and 26 cytokines and chemokines (pg/ml) were determined in cell-free supernatants. Results: In both experiments, human myeloid dendritic cells incubated with HagB produced Th1, Th2, and Th17 cytokines (IL-2, IL-12(p70), IFN-, IL-3, IL-4, IL-5, , IL-15, IL-17); pro-inflammatory cytokines (IL-1, IL-1, IL-6, TNF-, IL-12(p40); anti-inflammatory v cytokines (IL-10, IL-13, IFN2); chemokines (CXCL8/IL-8, CXCL10/IP-10, CCL2/MCP-1, CCL3/MIP-1, MIP-1b, CCL11/eotaxin); and colony stimulating factors (IL-7, G-CSF, GM-CSF). Histatin 5 significantly attenuated (p < 0.05) the IL-8 response induced by HagB at 8 - 16 hours and to a lesser extent, the IL-6, GM-CSF, MCP-1, MIP-1α, MIP-1β, and TNF-α response. Conclusion: Histatin 5 is an important salivary component capable of attenuating an IL-8 response. Together with human beta defensin 3, another peptide previously shown to attenuate pro-inflammatory cytokines, histatin 5 may help control and contain oral infection and inflammation by down regulating IL-8 chemotactic response.

Angiotensin II (Ang II) promotes vascular disease and hypertension, in part, by activating the interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) signaling pathway. Extensive studies have demonstrated that SOCS3 plays an important role in suppressing the IL-6/STAT3 pathway in the immune system and in cancer biology. In contrast, the functional importance of SOCS3 in cardiovascular disease is largely unknown. Thus, the overall goal of these studies was to investigate the role of SOCS3 in models of Ang II-dependent vascular disease and hypertension. To examine direct effects of Ang II on the vessel wall, carotid arteries from SOCS3 haplodeficient (SOCS3+/-) mice and wild-type littermates (SOCS3+/+) were incubated with the peptide or vehicle for 22 hrs, followed by examination of endothelial function using acetylcholine. Relaxation to acetylcholine was similar in all arteries incubated with vehicle. A low concentration of Ang II (1 nmol/L) did not affect acetylcholine-induced vasodilation in SOCS3+/+ mice, but reduced responses in arteries from SOCS3+/- mice by ~50% (P<0.05). This Ang II-induced endothelial dysfunction in SOCS3+/- mice was prevented by inhibitors of NF-êB or STAT3, an IL-6 neutralizing antibody, or a scavenger of superoxide. Responses to nitroprusside, an endothelium-independent vasodilator, were similar in all groups. To test the importance of SOCS3 in vivo, mice were infused systemically with a pressor dose of Ang II (1.4 mg/kg per day) or vehicle for 14 days via osmotic mini-pumps. Acetylcholine-induced vasodilation in carotid and resistance arteries in brain from SOCS3+/- mice was reduced by ~60% (P<0.05). Surprisingly, genetic deficiency in SOCS3 prevented the majority of Ang II-induced endothelial dysfunction without affecting the pressor response to Ang II. To investigate potential mechanisms underlying divergent results when studying effects of local versus systemic effects of Ang II, we performed bone marrow transplantation followed by infusion of vehicle or Ang II for two weeks. Lethally irradiated WT (CD45.1) mice reconstituted with SOCS3+/- bone marrow were protected from Ang II-induced endothelial dysfunction (P<0.05), while reconstitution of irradiated SOCS3+/- mice with WT (CD45.1) bone marrow exacerbated Ang II-induced vascular dysfunction (P<0.05). WT (CD45.1) into SOCS3+/+ and SOCS3+/- into SOCS3+/- bone marrow chimeras exhibited vascular function consistent with non-irradiated controls. In addition, the pressor response to Ang II was reduced by ~50% in WT mice reconstituted with bone marrow from SOCS3+/- mice (P<0.05). These data suggest that SOCS3 exerts divergent or context-dependent effects depending on whether vascular dysfunction was due to local versus systemic administration of Ang II. SOCS3 deficiency in the vessel wall enhanced local detrimental effects of Ang II on vascular function. In contrast, bone marrow-derived cells that are haplodeficient in SOCS3 protect against systemically administered Ang II and the resulting vascular dysfunction and hypertension. To my knowledge, these are the first experimental studies that begin to define the importance of SOCS3 in Ang II-induced hypertension and endothelial dysfunction. Results obtained from these experiments provide new insight into mechanisms which regulate oxidative stress and inflammation within the vasculature. The studies also revealed that bone marrow-derived cells that are haplodeficient in SOCS3 protect against pressor and endothelial effects of Ang II. These findings may eventually contribute to the development of novel therapeutic approaches for hypertension and hypertension associated end-organ damage.