Regensburg, Univ., Diss., 2009.

München, Techn. Univ., Diss., 2003.

Regensburg, Univ., Diss., 2004. / Erscheinungsjahr an der Haupttitelstelle: 2003

Würzburg, Univ., Diss., 2001. / Erscheinungsjahr an der Haupttitelstelle: 2001

Universiẗat, Diss., 2004--Leipzig.

Tierärztliche Hochsch., Diss., 2005--Hannover.

Proefschrift Universiteit Maastricht. / Met bibliogr., lit. opg. - Met samenvatting in het Nederlands.

Orientador: Márcia Regina Brochetto Braga / Banca: Regina Barretto Cicarelli / Banca: Frederico Gonzalez Colombo Arnoldi / Resumo: Polybia paulista ou popularmente "paulistinha" é uma vespa social da família Vespidae. Possui hábitos urbanos e grande ocorrência no Sudeste do Brasil, especialmente no Estado de São Paulo, onde tem causado muitos acidentes de importância médica. Na composição de seu veneno encontra-se um potente alérgeno, a proteína Antígeno 5 (Ag5) e embora sua função biológica ainda seja desconhecida, este alérgeno é responsável por importantes reações imunológicas cruzadas com o Ag5 do veneno de outros insetos sociais e com outras proteínas de eucariotos. A importância da reatividade cruzada em pacientes alérgicos ao veneno de vespas sociais é inquestionável, pois estas interações têm impacto direto sobre o diagnóstico e a seleção da melhor conduta terapêutica. Os diagnósticos de alergia são baseados na detecção de anticorpos do tipo IgE específico ao veneno por testes cutâneos ou de sangue. No entanto, respostas falso-positivas decorrentes da reatividade cruzada e respostas falso-negativas provenientes da baixa quantidade de IgE detectada, dificultam a interpretação dos resultados. A sequência completa de cDNA (621 pb) do alérgeno Ag5 do veneno da vespa P. paulista, foi clonada e a análise dos nucleotídeos revelou uma similaridade de 99% com a vespa Polybia scutellaris. Anticorpos policlonais foram produzidos contra a fração eletroforética protéica do Ag5 (25 kDA) de P. paulista e analisados imunologicamente por Western blotting. Os resultados demonstraram que os anticorpos reconheceram especificamente o alérgeno Ag5 no veneno bruto de P. paulista bem como, desenvolveram maior reação imunológica cruzada com os alérgenos Ag5 do veneno das vespas do gênero Polybia, embora não se descarte a possibilidade de ocorrência de reação cruzada com venenos de outros insetos sociais... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Polybia paulista, commonly known as "paulistinha", is a social wasp of the family Vespidae. This species occurs in urban areas and is frequent in the Southeastern Brazil, especially in the State of São Paulo, where it has been responsible for many accidents of medical significance. A potent allergen, the protein antigen 5 (Ag5), is an important compound of the wasp venom. Although its biological function remains unknown, this component is responsible for substantial cross-immunological reactions with the Ag5 of venoms of other social insects and with eukaryotic proteins. The importance of cross-reactivity in allergic patients to the wasp venoms is unquestionable, because these interactions present a direct impact on the diagnosis and on the selection of the therapeutic treatment. Allergenic diagnoses are based on the detection of IgE specific to the venom via cutaneous or blood tests. However, sometimes they are hampered by false-positive responses as a result of cross-reactivity and false-negative responses that can occur due to the low amount of IgE detected as consequence of the low sensitivity of the test. The fulllength cDNA (621 bp) from the venom allergen Ag5 wasp P. paulista was cloned and nucleotide analysis revealed 99% of similarity with the wasp Polybia scutellaris. Polyclonal antibodies were produced against the electrophoretic protein fraction of Ag5 (25 kDa) of P. paulista and immunologically analyzed by Western blotting. The results showed that the antibodies strongly recognized the allergen Ag5 in the venom of P. paulista and developed higher cross-immune reaction with the same allergen in wasp venoms of the genus Polybia, although the possibility of cross reaction with other insect venoms not tested in this study cannot be excluded. The model carried out for the Ag5 P. paulista revealed the... (Complete abstract click electronic access below) / Mestre

>Magister Scientiae - MSc / Prostate cancer is a deadly disease that occurs in the male’s prostate gland. A prostate gland is a walnut structure that forms part of the male’s reproductive system. Prostate cancer is caused by high level than normal of PSA (Gleason score > 4 ng ml-1) in human blood. Some symptoms associated with high levels of PSA include blood in urine, pain when urinating, difficulty in getting and keeping an erection, blood in semen and pain in upper thigh. An immunosensor is a type of biosensor that has an antigen or antibody fragment as its biological recognition component. The specificity of the molecular recognition of antigen by antibodies to form a stable complex is the basis of immunosensor technology. In this work, overoxidized polypyrrole (OvoxPpy) was electrosynthesized as a novel sensor platform on glassy carbon electrode (GCE). The OvoxPpy was then doped with osmium telluride quantum dots(OsTe2QDs) by drop-coating method to form OsTe2QDs|OvoxPpy|GCE system. The morphology and the size of OsTe2QDs|OvoxPpy|GCE nanocomposite were determined using scanning electron microscopy. The size of thioglycolic acid capped osmium telluride quantum dots (TGA-OsTe2QDs) used as support material for the biosensor was about 2.289 nm. These quantum dots showed an excellent photo-absorption properties with an ultraviolet- visible (UV-Vis) photo absortion band occurring at 406nm associated with high band energy of 3.05 eV. The electrochemical immunosensor for PSA was prepared by immobilizing anti- PSA-antibody onto the OsTe2QDs|OvoxPpy|GCE by drop-coating and allowing it to dry for 2h. The nanocomposite sensor platform and the immunosensor were electrochemically characterised by voltammetric and impedimetric techniques. The phase shift in Bode diagram at maximum frequency was indicative of kinetic changes. Charge transfer resistance, Rct, was used as the analytical parameter for measuring the interfacial kinetics which occurred as a result of the bio-recognition event between anti-PSA-antibody and PSA. The impedance of the quantum dot electrode (TGA-OsTe2QDs-Nafion|GCE) was lower (1.490 x 104 kΩ) than the impedance of the immunosensor platform (BSA-Anti-PSA-antibody|TGA-OsTe2 QDs|OvoxPpy|GCE), 2.754 x 104. The Rct of the immunosensor was found to increase with increasing concentration of PSA. The linearity of the immunosensor at the very low concentration range (1.266 - 4.207 fg ml-1) tested, confirms its high sensitivity for PSA.

The aims of this project were to investigate the effect of immune complexes on mononuclear phagocytes in the presence or absence of infection with the human immunodeficiency virus (HIV). Mono Mac 6 (MM6) cells and human monocyte-derived macrophages (MDM) were used to compare the effect of chemical stimulants, cytokines and immune complexes on surface antigen expression. The same experiments were performed using HIV-infected cells to determine the effect of HIV infection on these parameters. Some of the stimulants especially IL-6, IL-10, TNF-alpha and HIV increased CD80 expressions, the effect being greater in MDM than MM6 cells. This has implications for antigen presentation. Cytokines caused the differentiation of MM6 cells and significantly increased or decreased surface antigen expressions. The MM6 cells express surface antigens at lower levels then MDM this indicates that MM6 cells need to differentiate before expressing surface antigens. The high standard deviations obtained meant that no significant change in surface antigen expression was seen in all HIV non-infected and infected MDM incubated with various immune complexes and HIV-sera. Comparing whole blood incubated with KLH and rabbit anti-human IgG showed that both seemed to produce IL-10 and IL-6 early. Only rabbit complexes stimulated TNF-a release. MDM incubated with IL-6, IL-10, and TNF-alpha showed increased expression of CD16 and CD80 only at day 3. However, HIV proteins (CHO) incubated in whole blood caused a significantly release of IL-6 at 8 hours with no detection of IL-10 and TNF-alpha. MM6 infected with HIV-1 Ba-L for 5 days showed increased expression of CD16 and CD11c, but reduced expression of the other antigens examined. Significant levels of IL-10 were released at 8 and 12 hours with a slight increase in IL-6, and TNF-alpha. HIV protein-containing (CHO) immune complexes slightly increased IL-6 secretion at 8 hours at which point, IL-10 production was high. HIV- infected cells incubated with HlV-sera showed a lack of TNF-alpha release but IL-6 and IL-10 was detected at 12 and 8 hours respectively. The detection of high levels of IL-10 and the inhibition of TNF-alpha production may stimulate progression to full blown AIDS.