Thesis (M.S.)--University of Delaware, 2008. / Principal faculty advisor: Douglas W. Tallamy, Dept. of Entomology & Wildlife Ecology. Includes bibliographical references.

No description available.

Resumo: Não disponível / Abstract: Uncaria tomentosa (Willd.) DC (Rubiaceae) is a large woody vine that is native to the Amazon and Central American rainforests and is widely used in traditional medicine for its immunomodulatory and anti-inflammatory activities. The present work used in vivo immunotoxic and in vitro immunomodulatory experiments to investigate the effects of a pentacyclic oxindole alkaloid extract from U. tomentosa bark on lymphocyte phenotype, Th1/Th2 cytokine production, cellular proliferation and cytotoxicity. For the in vivo immunotoxicity testing, BALB/c male mice were treated once a day with 125, 500 or 1250 mg/kg of U. tomentosa extract for 28 days. For the in vitro protocol, lymphocytes were cultured with 10 to 500 μL/mL of the extract for 48 h. The extract increased the cellularity of splenic white pulp and the thymic medulla and increased the number of T helper lymphocytes and B lymphocytes. Also, a large stimulatory effect on lymphocyte viability was observed. However, mitogen-induced T lymphocyte proliferation was significantly inhibited at higher concentrations of U. tomentosa extract. Furthermore, an immunological polarization toward a Th2 cytokine profile was observed. These results suggest that the U. tomentosa aqueous-ethanol extract was not immunotoxic to mice and was able to modulate distinct patterns of the immune system in a dose-dependent manner / Orientador: Rosa Marlene Viero / Coorientador: Alexandrina Sartori / Banca: Carla Adriene da Silva Franchi / Banca: Ana Lúcia Tozzi-Spinardi Barbisan / Banca: Isis Machado Hueza / Banca: Efigênia Queiroz de Santana / Doutor

Sleep loss alters both the concentration and activity of various aspects of the immune system. These alterations lead to increased susceptibility to infection and the progression of pathologies such as insulin resistance and atherosclerosis. Two proposed mechanisms of this alteration in immune function are the changes in both cortisol and sympathetic nervous system activity that accompany sleep deprivation. This work reviewed literature that measured the effects of periods of sleep restriction upon both cortisol and catecholamine concentrations within human subjects. Furthermore, studies which measured the effects of sleep loss upon these hormone levels and the associated changes in immune parameters were included. This thesis asserts that there is no defined pattern in reference to alterations of cortisol levels as a result of sleep deprivation. Furthermore, more evidence must be collected before implementing cortisol as a main effector of sleep loss upon immune system function. This dissertation, although repeatedly noting increased levels of norepinephrine following periods of sleep restriction, similarly argues that more research must be completed in order to declare that altered catecholamine concentrations as a result of sleep loss is a mechanism for altered immune function.

It has been suggested that moderate exercise contributes to protection against the development of chronic diseases by anti-inflammatory mechanisms that include elevations of anti-inflammatory cytokines and also reduction of the expression of Toll- Like Receptors (TLRs). However, prolonged strenuous exercise has been shown to reduce the function of some immune cells, decrease virus protection and consequently may account for the reason athletes appear more vulnerable to catching Upper Respiratory Tract Infections (URTI). Although it has been proven that some exercise is better than no exercise, it is not clear yet what is the right amount of exercise to elicit beneficial immune responses and to help prevent the development of diseases. Therefore, the general aim of the studies in this thesis was to evaluate the impact of different types of exercise on monocyte TLR expression in participants with different fitness levels. It was found that different acute exercise protocols elicit different changes in TLR2 and TLR4 expression, where an acute bout of strenuous exercise reduced TLR4 expression for a few hours after the completion of the exercise (Chapter 5); however, short two bouts of exhaustive exercise separated by 2 hours did not change TLR4 expression (Chapter 6). In addition, changes in TLR4 expression were related to sex and the physical activity level of the participants (Chapter 4), and should therefore be considered separately when analysing TLR4 expression. Furthermore, high-intensity intermittent training improves participants' aerobic capacity and modifies the monocyte subpopulation concentration in the blood, with no changes in TLR4 expression. Further research needs to be done in this area to achieve a conclusive finding about changes in TLR4 expression and monocyte subsets after different training protocols, and possible relationships to cytokine production.

背景黃芪是中醫藥中最常用的補益藥之一，現代研究發現其多糖類成分具有抗疲勞、抗氧他和免疫調節等作用，因此研究黃芪多糖的生理活性已成為研究黃芪藥理作用新的主流方向。由於多糖類物質分子量較大，單糖組成及組成方式多樣，所以對多糖的研究除生理活性外還需探討解析其他學特征。目的初步驗證黃芪多糖對免疫系統的生理活性及其自身他學組成，探討展望未來對黃芪多糖研究的新方向。方法本實驗採用水提醇沉法提取分離除黃芪粗多糖，經除蛋白及透析等操作純忙得到黃百多糖。採用高效凝膠色譜分離法及超高效液相色譜法分別求得黃芪多糖相對分子量大小及其單糖組成成分免疫活性探究使用RAW264.7 細胞系巨噬細胞，以脂多糖為陽性對照，採用MTT 法測試細胞毒性，計算加藥后一氧化氮及細胞因子IL-6 和TNF－α 生成量，評價黃芪多糖的免疫調節作用。結果黃芪多糖相對分子量為108.02kDa(±2.73kDa），由阿拉伯糖、葡萄糖、半乳糖、葡萄糖醛酸和半乳糖醛酸組成。MTT 實驗表明黃芪多糖對細胞無明顯毒性﹔ NO 及細胞因子IL-6 和TNF－α 生成量表明其具有免疫調節功能，且作用強度與黃芪多糖濃度在一定範圍內呈正相關。結論黃芪多糖具有免疫調節活性，但其組成成分較多，他學結構複雜，仍需要進行更多研究探討其作用機制及其他學結構與免疫調節機制的關係。【關鍵詞】黃芪多糖﹔化學組成﹔免疫活性

Thesis (MTech (Biomedical Sciences))--Cape Peninsula University of Technology, 2017. / Type 2 diabetes mellitus (T2DM) is a chronic inflammatory condition characterised by hyperglycaemia; continuous activation of T-lymphocytes and immune dysregulation. Although the exact mechanisms of these phenomena are not fully understood, there is strong evidence suggesting the involvement of T-cells in the chronic inflammatory environment which could predispose diabetics to infections and thrombotic events. The effect of hyperglycaemia on cells of the innate immune system in T2DM has been well described and implicated in the progression of the disorder and the development of its complications. However, studies investigating the adaptive immune response still remain scarce and controversial. Thus, investigating T-cells in hyperglycaemic conditions could provide further insight into the immune dysfunction observed in T2DM and assist in identifying pathways which could be targeted in the disease management and treatment. Therefore, this study aimed to investigate chronic immune activation by measuring the expression of T-cell activation markers in hyperglycaemia and compare the results to those in the normoglycaemic group.

O nonilfenol etoxilado (NPE) é um desregulador endócrino que está presente no meio ambiente devido ao seu uso como detergente nos processos de limpeza de efluentes industriais. O objetivo deste trabalho foi avaliar a influência do NPE sobre o sistema imune adaptativo em ratos Wistar machos. Nestes animais foram avaliados o efeito do NPE sobre as células linfocitárias periféricas através da realização de hemograma e do perfil linfocitário adaptativo, analisando os marcadores de superfície CD4, CD8, CD28 e CD45 RA. Foi também avaliado o efeito do tratamento sobre o fígado e baço, bem como sobre o metabolismo intermediário, através das análises de glicemia, triglicerídeos e colesterol. Os dados não demonstraram diferenças significativas em relação ao índice hepático e esplênico. O nível de triglicerídeos apresentou um aumento de 50% nos grupos tratados, na avaliação dos níveis de colesterol e glicose não foi demonstrado diferenças significativas entre os grupos. Os resultados indicaram que o número de linfócitos e monócitos dos grupos tratados tiveram uma queda significativa de aproximadamente 25% e 50% em relação ao grupo controle. Foi demonstrado que o número de células fortemente marcadas quanto à presença da proteína CD45RA High na superfície celular dos linfócitos é maior nas células dos ratos do grupo tratado e que o tratamento aumenta a relação entre as células CD45RA High/Dim. Esses resultados levantam a hipótese que as células aumentadas nos grupos tratados apresentam fenótipo de membrana compatível com células T terminalmente diferenciadas (TEMRA). Este estudo forneceu dados novos sobre a ação do NPE, até onde se tem conhecimento, é a primeira pesquisa a constatar a presença elevada de células TEMRA em animais tratados com NPE, contribuindo com um novo foco para futuras pesquisas dessa substância. / Conselho Nacional de Desenvolvimento Científico e Tecnológico, CNPq / Ethoxylated nonylphenol (NPE) is an endocrine disruptor that is present in the environment because of its use as a detergent in the industrial effluent cleaning processes. The objective of this work was to evaluate the influence of ethoxylated nonylphenol (NPE) on the adaptive immune system in male Wistar rats. In these animals, the effect of NPE on peripheral lymphocyte cells was evaluate by performing hemogram and adaptive lymphocytic profile, analyzing CD4, CD8, CD28 and CD45 RA surface markers. The effect of treatment on the liver and spleen, as well as on the intermediate metabolism, was also evaluate through glycemic, triglyceride and cholesterol analyzes. The data did not show significant differences in relation to the hepatic and splenic index. The level of triglycerides presented a 50% increase in the treated groups; in the evaluation of cholesterol and glucose levels, no significant differences between the groups were demonstrate. The results indicated that both, the number of lymphocytes and monocytes of the treated groups had a significant decrease of approximately 25% and 50% relative to the control group. The number of strongly labeled cells for the presence of the CD45RA High protein on the cell surface of the lymphocytes showed to be higher in the cells of the mice in the treated group and that the treatment increases the ratio between the CD45RA High/Dim cells. These results raise the hypothesis that enlarged cells in the treated groups exhibit terminally differentiated T cell (TEMRA). This study provided new data on the action of NPE, to the best of our knowledge, is the first research to verify the elevated presence of TEMRA cells in animals treated with NPE. In addition, these findings contribute a new focus for future research on this substance.

Orientador: Renata de Oliveira Mattos-Graner / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-08T06:03:32Z (GMT). No. of bitstreams: 1 Nogueira_RucheleDias_D.pdf: 12321915 bytes, checksum: 624bf1e6309c440b0fb9bf9a2ed120f8 (MD5) Previous issue date: 2006 / Resumo: Streptococcus mutans (SM) são os principais patógenos da cárie dental. Neste estudo exploramos a influência do sistema imune de mucosa na infecção inicial pelo SM em crianças altamente expostas a este microrganismo. Cento e dezenove crianças, com idade inicial entre 5 a 13 meses de idade foram analisadas no início do estudo (T0) e após 6 (T6), 12 (T12) e 18 (T18) meses, amostras bucais foram coletadas para determinação dos níveis de infecção por SM, através do cultivo em mitis salivarius ágar com bacitracina. Os níveis de IgA, IgA1 e IgM foram também determinados em amostras de saliva, através de ensaios de ELISA. Um subgrupo de 21 crianças infectadas por SM (entre T0 e T6) foram pareadas a outras 21 crianças da amostra sem níveis detectáveis de SM. As reações entre anticorpos IgA da saliva com antígenos (Ags) de SM foram comparadas entre estes subgrupos, através de ensaios de western blot. A intensidade das bandas reativas foi determinada através da densitometria e expressas como unidades arbitrárias (ua). A reatividade de IgA salivar com peptídeos derivados da seqüência da GbpB, preditos como regiões ligantes à MHC de classe II, foram também avaliados através de multiplex com o Luminex. Os níveis salivares de IgA aumentaram com a idade (de 82,4 a 823,4µg/ml), enquanto que os níveis de IgM mantiveram-se baixos durante todo o estudo (média:4,2-2,8 µg/ml). Os níveis de infecção por SM foram maiores em T6 (média:45 ufc/área) e uma forte resposta de IgA aos Ags de SM pôde ser detectada logo aos 6 meses de idade. A resposta de IgA anti-GbpB foi observada em 38% das 21 crianças infectadas por SM, enquanto que 73% dos pares não infectados apresentavam estes anticorpos (Qui-quadrado, p<0.03). Um pico de resposta de IgA a GbpB ocorreu durante a fase de maior desafio de infecção (T6). Não houve diferenças significativas nos padrões de reatividade de IgA aos diversos peptídeos de GbpB testados, entre os grupos de crianças infectadas e não infectadas por SM. Os resultados indicam que uma resposta complexa de IgA a SM pode ocorrer a partir dos 6 meses e que a especificidade de resposta a Ags envolvidos na virulência podem influenciar na susceptibilidade à infecção por SM. Nenhum dos peptídeos da GbpB correspondeu às intensidades de resposta à GbpB nativa observadas entre os pares de crianças estudados / Abstract: Mutans streptococci (MS) are the main pathogens of dental caries. In this study we explored the influence of the muccosal immune system in the initial colonization by MS in children highly exposed to this microorganism. A total of 119 children, who were of 5 to 13 months of age, were enrolled in this prospective study. MS levels of infection were determined in cultures on mitis salivarius agar with bacitracin at baseline (T0), 6 (T6), 12 (T12) and 18 months (T18)-of follow-up. Saliva samples were also collected at all phases of the study for determination of the levels of antibodies IgA, IgA1 and IgM in ELISA assays. A subset of 21 MS-infected early between T0 and T6 were matched to other 21children, but who were not MS-infected. Patterns of IgA antibody reactivity to MS Ags were compared between these 21 pairs through assays of western blot. Intensities of IgAreactive bands were determined densitometrically and were expressed as arbitrary unit (au). Reactivities of salivary IgA antibodies to peptides derived from the GbpB sequence, which were predicted as having affinity to MHC of class II molecules were evaluated in multiplex with the help of the Luminex technology. Levels of antibody IgA in saliva increased with age (82.4-823.35 µg/ml), while levels of IgM remained at low levels (mean:4.2-2.78 µg/ml). The highest levels of MS infection in the infected group were observed at T6 (mean:45 cfu/plate). Robust responses to SM antigens were detected in children as early as 6 months of age. Only 38% of early-infected children carried IgA-reative GbpB while this antigen was recognized by the majority (73%) of children that were not infected by these microorganisms (chi-square, p<0.03). A peak of IgA response to GbpB occurred during the phase of highest SM infectious challenge (by T6). All the GbpB-derived peptides have shown reactivity with salivary IgA, independently of the status of MS infection. The results indicated that complex salivary IgA responses to MS Ags can occur by 6 months of age and that the patterns of salivary IgA specificities might influence in the susceptibility of initial infections to MS. Neither of the peptides tested corresponded to differences in response to the native protein observed between the pairs of children studied. / Doutorado / Microbiologia e Imunologia / Doutor em Biologia Buco-Dental

Orientador: Margareth Castro Ozelo / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-15T17:29:43Z (GMT). No. of bitstreams: 1 Santos_Andreydos_D.pdf: 4770033 bytes, checksum: 92cd7a4adf25e169140e6fbbb68fe9d2 (MD5) Previous issue date: 2010 / Resumo: Uma complicação decorrente do tratamento da hemofilia é a formação de anticorpos neutralizadores da atividade coagulante do fator VIII ou IX (inibidores). Diversos fatores estão relacionados com o desenvolvimento desses inibidores em indivíduos com hemofilia, incluindo fatores genéticos e ambientais. Entre os fatores genéticos, a mutação associada ao diagnóstico da hemofilia é um fator de risco bem documentado. Recentemente foi observada a maior ocorrência de inibidores em indivíduos da etnia negra. O objetivo deste trabalho foi analisar os aspectos genéticos e não genéticos envolvidos no desenvolvimento de inibidores. Foram incluídos nesse estudo 411 pacientes hemofílicos, sendo 321 com hemofilia A (HA) (238 famílias) e 99 com hemofilia B (HB) (59 famílias). A presença de inibidores foi constatada apenas entre os pacientes HA graves. Do total de 220 HA graves desse estudo, 46 (20,9%) apresentaram inibidor detectado em pelo menos uma ocasião após sua inclusão no estudo. Mutações consideradas de alto-risco para o desenvolvimento de inibidores foram identificadas em 125/220 pacientes HA graves (58,8%), e 33 deles desenvolveram inibidores (26,4%). Considerando o grupo étnico de acordo com traços físicos e ancestralidade, 38% dos pacientes HA graves foram classificados como negros. A incidência de inibidores foi maior nesse grupo de pacientes (31% do total de pacientes HA graves classificados como negros) quando comparada aos pacientes caucasóides (20% do total de pacientes HA graves classificados como caucasóides). Recentemente, foi observado que a maior incidência de inibidores em uma população norte-americana de pacientes com HA, estava relacionada com a presença de determinados haplótipos no gene do fator VIII. Esta observação poderia ser explicada pelo fato dos as proteínas expressas pelos haplótipos que aparecem exclusivamente entre a população negra (denominados H3 e H4), estarem ausentes nos concentrados de fator VIII recombinantes utilizados rotineiramente no tratamento desses pacientes. Em nossa análise a presença desses haplótipos não está relacionada com a maior freqüência de inibidor na população negra desse estudo. Além disso, a distribuição dos diferentes haplótipos do gene do fator VIII, classificados de H1 a H6, foi distinta entre todos os grupos étnicos brasileiros e norte-americanos. Essa observação pode ser explicada pela origem distinta entre os negros que imigraram da África para o Brasil e para a América do Norte, assim como o alto índice de miscigenação de nossa população. Em outra fase desse estudo, foi realizada a análise comparativa da expressão gênica a partir de amostras de RNA mensageiro (RNAm) extraídas em pool leucocitário de pacientes com HA grave, com ou sem a presença de inibidores. Na avaliação que incluiu numa primeira análise pacientes de uma mesma família discordante para a presença de inibidor e, em uma segunda fase, indivíduos não relacionados, foram observados 50 genes mais expressos e 16 genes menos expressos em pacientes com inibidor em comparação aos sem inibidor. Dentre esses genes foram selecionados dez, levando-se em conta sua participação na resposta imune ou sua correlação prévia com o desenvolvimento de inibidores em outros estudos. Pela técnica de PCR em tempo real, observou-se que os genes da interleucina 8 (IL-8) e da cistatina F (CST7) demonstraram ser mais expressos em pacientes com inibidor, enquanto que o gene da interleucina 10 (IL-10) foi menos expresso nesse grupo de pacientes. Dessa forma, nossos resultados fortalecem a idéia de que o mecanismo de desenvolvimento de inibidores em hemofilia é complexo e ainda não totalmente esclarecido e que existe um grande envolvimento de diversos genes relacionados com sistema imune na formação desses inibidores. O estudo em diferentes populações é uma importante etapa para o entendimento dos fatores de risco para o desenvolvimento de inibidores. Esse foi o primeiro trabalho realizado no Brasil incluindo pacientes de diversas regiões e analisando simultaneamente diferentes fatores e seu envolvimento com o desenvolvimento de inibidores. A determinação desses fatores de risco ajudará no futuro a determinar um tratamento diferenciado para o controle e sobretudo prevenção do desenvolvimento de inibidores / Abstract: The most serious complication of the treatment of hemophilia is the development of neutralizing antibodies to coagulation activity of factor VIII or IX (inhibitors). Several risk factors are related to the development of these inhibitors in patients with hemophilia, including genetic and environmental factors. Among genetic factors, the mutation associated with the diagnosis of hemophilia is a risk factor well documented. Recently, it was observed a higher incidence of inhibitors in African ancestry patients. The aim of this study was to analyze the genetic and non-genetic factors involved in the development of inhibitors. The study included 411 hemophilia patients, of which 321 with hemophilia A (HA) and 99 with hemophilia B (HB), belonging to a total of 238 and 59 families, respectively. The inhibitors were observed only in severe HA patients. From the 220 severe HA, 46 (20.9%) had inhibitor. The high risk mutation for the development of inhibitors were identified in 125 / 220 (58.8 %) severe HA patients, and 33 (26.4 %) of them developed inhibitors. Considering the ethnic group according to physical traits and ancestry, 38 % of severe HA patients were classified as black. The incidence of inhibitors is higher in this group of patients (31%) when compared to Caucasian patients (20%). The higher risk of inhibitor among African-Brazilians, could not be explained by the presence of the distinct factor VIII haplotypes, such as H3 and H4, as suggested in previous study. In fact, the prevalence rates of these haplotypes were distinct between Brazilians and North Americans, probably due to the fact that migrations of blacks to Brazil and to North America were originated from different geographic areas of Africa. In another phase of this study, we performed a comparative analysis of gene expression in samples of messenger RNA (mRNA) extracted from leukocytes of inhibitor and non-inhibitor patients with severe HA was performed. The evaluation consisted of an initial analysis of severe HA patients siblings, or from the same family, discordant for inhibitor development and in a second phase a group of unrelated individuals. Using the bioarrays technology 50 genes were upregulated and 16 were downregulated in inhibitor patients compared with non-inhibitor patients. Ten genes were selected among them, which are involved in immune response and were related to inhibitors development in other studies. It was observed by real time PCR that the genes for interleukin 8 (IL-8) and cystatin F (CST7) were upregulated and for interleukin 10 (IL-10) was downregulated in inhibitor patients. In conclusion, our results strengthen the idea that the mechanism of inhibitor development in hemophilia is complex, not clear and there is a large involvement of several genes related to the immune system in the development of these inhibitors. The study in different populations is important to understand the risk factors for the development of inhibitors. This is the first work in Brazil, to study patients from various regions and to performe analysis of different factors and their involvement in the development of inhibitors. The determination of these risk factors will help in the future to determine differential treatment for the control and in particular, for preventing the development of inhibitors / Doutorado / Clinica Medica / Doutor em Clínica Médica