Made available in DSpace on 2014-06-11T19:33:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-09-16Bitstream added on 2014-06-13T21:06:11Z : No. of bitstreams: 1 melo_rf_dr_arafo.pdf: 606373 bytes, checksum: 12056dceba1a7f6dbd020e29b732d781 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O objetivo deste estudo foi o de avaliar possíveis interações clínicas, microbiológicas, imunológicas e genéticas que possam influenciar o sucesso de implantes osseointegrados. Foram avaliados 47 implantes, em 47 pacientes, sendo 31 em condições de saúde (G I) e 16 com perimplantite (G II) e 47 dentes, dos quais 31 estavam sadios e eram dos pacientes com implantes sadios (G III) e 16 dentes sadios dos pacientes com perimplantite (G IV). Foi realizado exame clínico completo em todos os implantes e dentes selecionados. Amostras de fluido crevicular perimplantar/gengival do sítio com maior profundidade de sondagem foram coletadas. A avaliação das bactérias A. actinomycetemcomitans, P. gingivalis, P. intermedia, P. nigrescens e T. forsythia foi realizada pela técnica de PCR e, a quantificação das citocinas IL-1β e IL-6 foi realizada pelo teste ELISA. Células da mucosa bucal foram coletadas para avaliação dos polimorfismos IL1B+3954, IL1B-511 e IL6-174. A avaliação estatística dos parâmetros clínicos SS, SUP, PS e NI revelaram que os implantes do grupo G II apresentaram piores condições clínicas em comparação ao grupo G I. O grupo G II também apresentou piores condições clínicas que o grupo G IV para PS e NI. A análise microbiológica revelou que a bactéria A. actinomcetemycomitans não estava presente em nenhum sítio avaliado. P. intermedia também não foi encontrada no grupo G II. As bactérias estudadas apresentaram proporções semelhantes em todos os grupos avaliados, não havendo diferença entre os grupos. Na análise da concentração de IL-1β e IL-6, não houve diferenças significativas entre os grupos. A população estudada está em Equilíbrio de Hardy-Weinberg. Os polimorfismos estudados não demonstraram predominância dos alelos e dos genótipos. Nenhum polimorfismo foi associado à condição de doença. / The aim of the present study was to evaluate clinical, microbiological, immunological and genetics parameters in patients with implant loaded at least for one year. It was examined 47 implants and teeth in 47 patients. Thirty one of those implants were healthy implants (G I), sixteen had peri-implantits (G II) and, 31 healthy teeth was from patients with healthy implants (G III) and 16 healthy teeth from patients with peri-implantits (G IV). Clinical parameters were recorded from all implants and teeth. Gingival crevicular fluid from the highest pocket depth was collected to evaluate the presence of A. actinomycetemcomitans, P. gingivalis, P. intermedia, P. nigrescens e T. ForsythiaI and to evaluate the concentration of interleukin-1β and interleukin-6. Cells from buccal mucosa were collected for genomic DNA extraction and analyze the polymorphism IL-1B +3954, IL-1B -511 e IL-6 -174. G II demonstrated worst results for PD, BPD, Sup and NI when compared to G I and, when compared with G IV it was worst for PD and NI. Microbiological did not detect A. actinomycetemcomitans in any of the sites analysed and, P. intermedia was not detected in G II. Bacteria analyzed was present in same proportion in all analyzed sites showing, no differences between groups. There was no difference in the concentration of IL-1β an IL-6 detected between groups. The population studied was in Hardy-Weinberg Equilibrium. There was no differences in the alleles and polymorphism distribution on the studied population.

CoordenaÃÃo de AperfeÃoamento de Pessoal de NÃvel Superior / O objetivo deste estudo foi investigar a expressÃo do sistema interleucina 1 (proteÃnas e RNAm de ligantes e receptores) e sua distribuiÃÃo nos ovÃrios de vacas cÃclicas, bem como avaliar os efeitos da IL-1β na sobrevivÃncia e ativaÃÃo de folÃculos primordiais in vitro. Os ovÃrios foram processados para a localizaÃÃo do sistema interleucina 1 em folÃculos prÃ-antrais e antrais utilizando as tÃcnicas de imunohistoquÃmica, qPCR e anÃlise de Western blot. Para os estudos in vitro, fragmentos ovarianos foram cultivados em α-MEM+ suplementado com IL-1β (0, 1, 10, 50 ou 100 ng/mL), e apÃs 6 dias foram processados para anÃlise histolÃgica. Os resultados de imunohistoquÃmica mostraram que a proteÃna para os integrantes do sistema interleucina I (IL-1β, IL-1RA, IL-1RI e IL-1RII) foram detectados em diferentes compartimentos foliculares. Infelizmente, o anticorpo testado para localizaÃÃo de IL-1α nÃo reagiu em ovÃrios bovinos. Todas as proteÃnas testadas foram observados no citoplasma dos oÃcitos e nas cÃlulas da granulosa de todas as categorias foliculares, e nas cÃlulas da teca de folÃculos antrais, com a exceÃÃo da IL-1α, que nÃo foi encontrada em nenhuma das cÃlula analisados. Foram observados nÃveis variÃveis de RNAm para o sistema interleucina 1 nas diferentes categorias foliculares analisadas. ApÃs 6 dias de cultivo, a presenÃa de IL-1β (10 ou 50 ng/mL) foi capaz de manter a percentagem de folÃculos normais e de promover a ativaÃÃo dos folÃculos primordiais. Em conclusÃo, os componentes do sistema de interleucina 1 sÃo expressos diferencialmente em cÃlulas ovarianas de acordo com a fase do desenvolvimento folicular. AlÃm disso, a IL-1β promove o desenvolvimento de folÃculos primordiais in vitro. Estes resultados sugerem um importante papel do sistema interleucina 1 na regulaÃÃo da foliculogÃnese em bovinos. / This study aims to investigate the expression of interleukin 1 system (proteins and mRNA of ligands and receptors) and its distribution in ovaries of cyclic cows, as well as to evaluate the effects of IL-1β on the survival and activation of primordial follicles in vitro. The ovaries were processed for localization of interleukin 1 system in preantral and antral follicles by immunohistochemical, qPCR and western blot analysis. For in vitro studies, ovarian fragments were cultured in α-MEM+ supplemented with IL-1β (0, 1, 10, 50 or 100 ng/mL), and aftes 6 days the tissues cultured were processed for histological analysis. Immunohistochemical results showed that the proteins for interleukin 1 system (IL-1β, IL-1RA, IL-1RI and IL-1RII) were detected in the various follicular compartments. Unfortunately, IL-1α antibodies tested did not react in bovine ovaries. All the proteins tested were observed in the cytoplasm of oocytes and granulosa cells from all follicular categories, and theca cells of antral follicles, with the exception that IL-1α has not been found in any analyzed cell. Variable levels of mRNA for the interleukin 1 system in the follicular size and classes analysed. After 6 days of culture, the presence of IL-1β (10 or 50 ng/mL) was effective in maintaining the percentage of normal follicles and in promoting primordial follicle activation. In conclusion, interleukin 1 system is differentially expressed in the ovarian cells according to the stage of follicular development. Moreover, IL-1β promotes the development of primordial follicles. These results suggest an important role of the interleukin 1 system in the regulation of folliculogenesis in bovine species.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A dor neuropÃtica possui mecanismos de difÃcil compreensÃo que se modificam com a sua evoluÃÃo dificultando a terapÃutica. O modelo por constriÃÃo crÃnica do nervo ciÃtico (CCNC) simula esta condiÃÃo. A gabapentina (GBP) à um anticonvulsivo muito utilizado como analgÃsico na dor neuropÃtica e existem poucos relatos sobre seus efeitos na atividade inflamatÃria e regenerativa do nervo. As citocinas TNF-&#945; e IL-1&#946; estÃo relacionadas à dor, inflamaÃÃo e regeneraÃÃo neural. à objetivo deste estudo avaliar os efeitos antinociceptivos, inflamatÃrios e regenerativos do tratamento preventivo e contÃnuo (80 min antes da CCNC e depois ao longo de 5 dias de 12 em 12h) com a GBP (30, 60, 120 mg/kg, via oral) atravÃs da observaÃÃo (antes, no 3 dia e no 5 dia apÃs a CCNC) dos seguintes comportamentos: (1) espontÃneos sugestivos de dor crÃnica: Scrathing (coÃar-se) e Biting(morder-se) (2) relacionados à motriciadade e exploraÃÃo: Climbing(escalar), Rearing(empinar-se), Walking(andando) (3) de dor induzida: alodÃnia mecÃnica (von Frey eletrÃnico) e alodÃnia ao frio (10Â) em ratos machos Wistar (250-300g) com CCNC. Animais com CCNC tratados com salina e animais pseudo-operados foram utilizados como controles. Para avaliar os efeitos anti ou prÃ-inflamatÃrios da GBP (60, 120mg/kg, via oral) utilizou-se o modelo de edema de pata induzido por carragenina em animais sem CCNC e a dosagem de mieloperoxidase no nervo sob constriÃÃo crÃnica. Os efeitos neuro-regenerativos da GBP foram avaliados indiretamente atravÃs da expressÃo de TNF-&#945;, IL-1&#946;, IL-10, e da proteÃna bÃsica de mielina (PBM) no mesmo protocolo, sendo que no 5 dia de tratamento apÃs CCNC, foram feitas coletas dos nervos para dosagem destas citocinas atravÃs de ELISA e imunohistoquÃmica, e no 5 e 15 dia para expressÃo de PBM atravÃs de Western Blot e imunohistoquÃmica. Resultados: No 5 dia, as doses de GBP 30, 60 e 120mg provocaram reduÃÃo de 86,76%*; 83,82%*; 80,09%* (*p < 0,01) no tempo de Scratching da pata ipslateral e uma inibiÃÃo de 56,27%; 90,53%*; 65,06% (*p < 0,01) no tempo do Biting lado direito em relaÃÃo ao grupo salina. Aliviaram a alodÃnia mecÃnica provocando aumentos do limiar de retirada da pata de 225,42%*, 246,01%*, 309,20%* (*p <0,001) e provocaram reduÃÃo do escore de dor na alodÃnia ao frio em 18,19%; 21,29%; 42,26%* (*p < 0,01) em relaÃÃo ao grupo salina. As doses de GBP 60 e 120mg provocaram aumentos significativos na motricidade espontÃnea de animais com CCNC e intactos sugerindo efeitos estimulantes e nÃo sedativos. A GBP 60mg/kg tambÃm provocou efeitos relevantes (p<0,001) na migraÃÃo de cÃlulas visto pelo aumento de 982,16% na expressÃo da mieloperoxidase no nervo e pelo aumento de 53% no edema de pata em relaÃÃo ao controle salina. Quanto à expressÃo do TNF-&#945; no nervo as doses de 60 e 120mg provocaram aumentos de 120,95 % e de 91,99% em relaÃÃo ao grupo salina (P< 0,01). Na expressÃo de IL-1&#946; a dose de 60mg/kg provocou aumento de 59,26% em relaÃÃo ao grupo salina (P< 0,05). No Western Blot ocorreu um aumento significativo de 158,06% da expressÃo de PBM (0,298 + 0,069 PBM/Act.) no nervo no 15 dia de tratamento com GBP em relaÃÃo ao grupo salina (0,116 + 0,034 PBM/Act). ConclusÃes: O tratamento com GBP ao longo de cinco dias, apesar de seus efeitos prÃ-inflamatÃrios, demonstrou no 5 dia efeitos analgÃsicos significativos ao reduzir comportamentos espontÃneos de dor crÃnica e aliviar a alodÃnia mecÃnica e tÃrmica em estÃgio precoce da neuropatia. Considerando-se os efeitos relevantes da GBP no 5 dia de neuropatia no aumento da migraÃÃo de cÃlulas para o local da lesÃo, no aumento da expressÃo de TNF e IL-1&#946; no nervo, o que levaria a maior ativaÃÃo das cÃlulas de Schwann, macrÃfagos, e maior liberaÃÃo de fator de crescimento do nervo, e no aumento da expressÃo de PBM indicando uma ativa fragmentaÃÃo da mielina, sugere-se que a GBP pode acelerar o processo de remoÃÃo dos fragmentos de mielina e axÃnios contribuindo assim para o posterior alongamento dos axÃnios e remielinizaÃÃo. / Neurophatic pain conveys mechanisms of difficult understanding, which changes with the disease progress, jeopardizing the therapeutics. The chronic constriction of the sciatic nerve (CCSN) model mimics this condition. Gabapentin (GBP) is an anti-convulsive drug often used as analgesics to control neurophatic pain. Few reports exist on their effects in the inflammatory and regenerative activity of the nerve. TNF-&#945; and IL-1&#946; cytokines, although related to pain and inflammation, are also associated with early myelin and axonal fragmentos removal after nerve injury and thus contribute to nerve regeneration. This study aimed at evaluating the analgesics early treatment (80 min prior to CCSN and along 5-day treatment, 12h/12h) effects of GBP (30, 60, 120 mg/kg, given orally) by monitoring (before surgery, on the 3rd day and 5th day of post-surgery treatment, 12h to12h) the following behaviors (1) spontaneous behavior, suggestive of chronic pain: Scratching and Biting (2) motor and exploratory-related behaviors: Climbing, Rearing, and Walking, (3) induced-pain behaviors: mechanical allodynia (electronic von Frey) and cold allodynia (10ÂC) in Wistar male rats (250-300g) with CCSN. CCSN animals treated with saline and pseudo-operated were used as controls. Another aim of this study was to evaluate the pro and anti-inflammatory effects of GBP (60 and 120mg/kg, given orally) by carragenin-induced paw edema in CCSN-free animals and yet nerve myeloperoxidase measurements under chronic constriction. The neural regenerative effects of GBP were indirectly evaluated by measuring TNF-&#945;, IL-1&#946;, IL-10, and myelin basic protein (PBM) expressions in the same protocol, as follows: 5th day of treatment following CCSN to draw nerves in order to quantify cytokines by ELISA and immunohistochemistry, and for MPB expression detected by Western blot and immunohistochemistry on the 5th and 15th. Results: On the 5th day, GBP, at doses of 30, 60 and 120mg, reduced the scratching time in the ipslateral paw of 86.76%*; 83.82%*; 80.09%* (*p<0.01) and an inhibition of the biting time of 56.27%; 90.53%*; 65.06% (*p<0.01) in the right side in comparison with the saline group. In addition, GBP alleviated mechanical allodynia causing increase paw retrieval thresholds of 225.42%*, 246.01%*, 309.20%* (*p <0.001) and the reduction of the pain scores for cold allodynia in the range of 18.19%; 21.29%; 42.26%* (*p < 0.01) in comparison with the saline group. GBP, at doses of 60 and 120mg, significantly increased spontaneous motricity in CCSN animals and controls suggesting excitatory effects rather than sedation. GBP (60 mg/kg) also caused relevant effects (p<0.001) in peritoneal cell migration, and a 982.16% increase in nerve myeloperoxidase expression and 53% increase in paw edema, as opposed to the saline control. Regarding nerve TNF-&#945; expression, doses of 60 and 120mg caused significant increase of 120.95 % and 91.99% in comparison with the saline group (p<0.01). A dose of 60mg/kg caused 59.26% increase in IL-1&#946; expression compared with the saline group (p<0.05). We found a 158% significant increase in PBM expression by western blots (0.298  0.069 PBM/Act) on the 15th treatment-day with GPB in comparison with the saline control (0.116  0.034 PBM/Act). Conclusions: 5-day GBP treatment, although with pro-inflammatory effects, was found to have significant pain killing effects by reducing chronic spontaneous pain behaviors and by alleviating mechanical and thermal allodynia in the early neuropathy course. By taking our relevant findings of increasing migration cell rates to the nerve injury site on the 5th day of the neuropathy and the nerve TNF and IL-1&#946; expression with more macrophage and Schwann cell activation and supposedly more nerve growth factor release, along with higher PBM expression, altogether suggest that GBP may hasten the myelin and axonal fragmentos withdrawn, therefore contributing to axonal outgrowth and ultimately remyelinization.

A periodontite agressiva (PAg) compreende um grupo de doenças periodontais raras caracterizadas por rápida destruição dos tecidos periodontais, em indivíduos jovens e que geralmente não apresentam doenças sistêmicas. Estudos em populações e em famílias indicaram que fatores genéticos possuem influência na susceptibilidade a periodontite agressiva. Os polimorfismos genéticos da interleucina-1 (IL-1) e do fator de necrose tumoral-? (TNF-?) foram associados com o aumento da severidade da periodontite crônica. O objetivo deste estudo foi avaliar a associação entre o polimorfismo dos genes IL-1A (-889) e TNFA (-308) e a periodontite agressiva. Foram selecionados 60 indivíduos não fumantes, sendo 30 portadores de periodontite agressiva e os outros 30 sem doença periodontal. O polimorfismo genético foi analisado utilizando-se a técnica da reação em cadeia da polimerase e análise do polimorfismo de comprimento dos fragmentos de restrição (PCR-RFLP). Foi observado que a freqüência do genótipo 1/1 para IL-1A foi de 63,3% no grupo controle e de 56,7% no grupo teste. A avaliação do genótipo 1/2 mostrou uma freqüência de 26,7% no grupo controle e de 40% no grupo teste. O genótipo 2/2 ocorreu com uma freqüência de 10% no grupo controle e de 3,3% no grupo teste. O genótipo 1/1 para TNFA estava presente em 73,3% do grupo controle e em 80% do grupo teste. O genótipo 1/2 ocorreu com freqüência de 20% em ambos os grupos. O genótipo 2/2 foi encontrado em 6,7% dos controles e não foi detectado no grupo teste. Em relação aos alelos, o alelo 1 apresentou freqüência de 76,7% e o alelo 2 de 23,3% para ambos os grupos, para o gene IL-1A (-889). E para o gene TNFA (- 308) o alelo 1 ocorreu com freqüência de 83,3% no grupo controle e de 90% no grupo teste e o alelo 2 de 16,7% no grupo controle e 10% no grupo teste. A análise estatística revelou que não houve diferença significativa na distribuição dos genótipos, para ambos os genes, entre os dois grupos estudados. Não foi encontrada associação entre a periodontite agressiva e o polimorfismo dos genes IL- 1A (-889) e TNFA (-308) na população estudada. / Agressive periodontitis (AgP) is a relatively uncommon form of periodontal disease characterized by a rapid destruction of the periodontal supporting tissues in young adults who are usually systemically well. The results of population and family studies indicate that genetic factors seem to have a strong influence on susceptibility to AP. Genetic polymorphism at the interleukin-1 (IL-1) and tumor necrosis factor alpha (TNFA) were associated with the increase on the severity of chronic periodontitis. The aim of this study was to explore a possible association between IL-1A and TNFA genotypes in Brazilian white Caucasian patients with aggressive periodontitis. Sixty nonsmoking subjects, 30 patients and 30 periodontal healthy controls were included in the study. All subjects were systemically healthy. Two polymorphisms, IL-1A (-889) and TNFA (-308), were analyzed by means of polymerase chain reaction-restriction fragment length polymorphism. The 1/1 genotype for IL-1A was present in 63.3% of the controls and in 56.7% of the aggressive periodontitis patients. The genotype 1/2 was present in 26.7% of the controls and in 40% of the patients. The 2/2 genotype was present in 10% of the controls and in 3.3% of the diseased subjects. The 1/1 genotype for the TNFA was present in 73.3% of the controls and in 80% of the patients. The genotype 1/2 was present in the 20% of both groups. The genotype 2/2 was present in 6.7% of the controls and was not detected in the patients group. With regard to the IL-1A (-889) genotype, 76.7% of controls and patients were positive for allele 1. Allele 1 of the TNFA (-308) polymorphism was carried by 83.3% of the controls and 90% of the patients and allele 2 was carried by 16.7% of the controls and 10% of the patients. Statistical analysis revealed no significant difference in the distribution of genotypes for both genes between the two groups. No association was found between AgP and the IL-1A (-889) and TNFA (-308) polymorphisms investigated in the population presented here.

Orientador: Rebeca de Souza Azevedo / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-23T13:31:17Z (GMT). No. of bitstreams: 1 Messetti_AnaCamilaPereira_M.pdf: 2046118 bytes, checksum: 6767fe84e5b25d194bac897de86b23a3 (MD5) Previous issue date: 2013 / Resumo: A doença periodontal (DP) é uma doença inflamatória crônica de ampla distribuição etária e mundial, que resulta da interação entre fatores etiológicos heterogêneos e que pode sofrer influência ambiental e sistêmica, especialmente na presença do diabetes melito (DM). Os mecanismos que envolvem a associação entre o DM e a presença da DP ainda não estão completamente elucidados, mas as características genéticas ou epigenéticas do hospedeiro, como os polimorfismos genéticos, podem influenciar esta relação. Dessa forma, o objetivo deste estudo foi avaliar o envolvimento de polimorfismos nos genes de IL1-? (-889), de IL1-? (+3984), de IL-6 (-174), de TNF-? (-308) e da paraoxanase (-192) e da paraoxonase (-55) no desenvolvimento de DP e DM em um grupo de pacientes brasileiros. Os polimorfismos rs1800587 e rs1143634 no gene IL-1, rs1800796 no gene IL6, rs1800629 no gene TNF-?, e rs662 e rs854560 no gene paraoxanase foram genotipados em 302 adultos, dos quais 96 eram diabéticos com DP (ADM+DP), 20 eram diabéticos com saúde periodontal (ADM-DP), 112 eram normoglicêmicos com DP (AN+DP) e 74 eram normoglicêmicos com saúde periodontal (AN-DP); e em 88 crianças e adolescentes, dos quais 11 eram diabéticos com gengivite (CDM+G), 15 eram diabéticos com saúde gengival (CDM-G), 30 eram normoglicêmicos com gengivite (CN+G) e 32 eram normoglicêmicos com saúde gengival (CN-G). No grupo de pacientes adultos, a presença do alelo T no polimorfismo rs1143634 no gene IL1-? foi mais frequente nos indivíduos ADM+DP que nos indivíduos AN-DP, gerando um OR de 14,9 (95% IC: 8,6-25,9; p<0,0001); a presença do alelo A no polimorfismo rs1800629 no gene TNF-? foi mais frequente nos indivíduos ADM+DP que nos indivíduos AN-DP, gerando um OR de 22,7 (95% IC: 6,96 -74,5 p<0,0001), e a presença do alelo C no polimorfismo rs1800796 no gene IL6 foi mais frequente nos indivíduos ADM+DP que nos indivíduos AN+DP, gerando um risco de recorrência de 2,38 (95% IC:1,61-3,5; p<0,0001), e que nos indivíduos AN-DP, gerando risco de recorrência de 2,87 (1,79-4,5; p<0,0001). No grupo de pacientes crianças e adolescentes, a presença do alelo C no polimorfismo rs1800796 no gene IL-6 foi mais frequente em indivíduos CN+G que nos indivíduos CN-G, gerando um risco de recorrência de 121,1 (95% IC:32,3-145,3; p<0,0001), e foi mais frequente nos indivíduos CDM+G que nos indivíduos CN-G, gerando um risco de recorrência de 104 (95% IC:11,4-136,6; p<0,0001). Os resultados deste estudo evidenciam a associação dos polimorfismos rs1143634 no gene IL1-?, rs1800796 no gene IL6 e rs1800629 no gene TNF-? com a suscetibilidade genética ao desenvolvimento de DP e/ou DM na população adulta estudada; e a associação do polimorfismo rs1800796 no gene IL-6 com a suscetibilidade genética ao desenvolvimento de DP e/ou DM na população de crianças e adolescentes estudada / Abstract: Periodontal disease (PD) is a chronic inflammatory disease of worldwide distribution that may affect all age groups, and results from the interaction between heterogeneous etiologic factors, such as environmental and systemic influence, especially in the presence of diabetes mellitus (DM). Mechanisms involving the association between DM and the presence of PD are still not fully elucidated, but the genetic or epigenetic alterations of the host, such as the genetic polymorphisms may influence this relationship. Thus, the aim of this study was to evaluate the involvement of polymorphisms in the genes of IL1-? (-889), IL1- (+3984), IL-6 (-174), TNF-? (-308) and paraoxanase (-192) and paraoxonase (-55) in PD and/or DM development in a group of Brazilian patients. The polymorphisms rs1800587 and rs1143634 from IL1 gene, rs1800796 form IL6 gene, rs1800629 from TNF-? gene, and rs662 and rs854560 from paraoxanase gene were genotyped in 302 adults, of whom 96 were diabetic with PD (ADM+PD), 20 were diabetics with periodontal health (ADM-PD), 112 were normoglycemic with PD (AN+PD) and 74 were normoglycemic with periodontal health (AN-PD); and in 88 children and adolescents, of whom 11 were diabetic patients with gingivitis (CDM+G), 15 were diabetics with gingival health (CDM-G), 30 were normoglycemic with gingivitis (CN+G) and 32 were normoglycemic with gingival health (CN-G). In the group of adult patients, the presence of T allele at the rs1143634 polymorphism in the IL1-? gene was more frequent in ADM+PD than in AN-PD, generating an OR of 14, 9 (95% CI :14,9-25, 9; p <0.0001), the presence of A allele at the rs1800629 polymorphism in the TNF-? gene was more frequent in ADM+PD than in AN+PD, generating an OR of 22,7 (95% CI 6.96 -74.5; p <0.0001), and the presence of C allele at the rs1800796 polymorphism in the gene IL6 was more frequent in ADM+PD than in AN+PD, generating an OR of 2.38 (95% CI :1,61-3, 5; p <0.0001), and it was more frequent in ADM+PD than in AN-PD, generating an OR of 2.87 (1.79 to 4.5, p <0.0001). In the group of children and adolescents, the presence of C allele at the rs1800796 polymorphism in the IL6 gene is more frequent in NC+G than in NC-G, generating an OR of 121.1 (95% CI :32,3-145, 3; p <0.0001), and it was more frequent in CMD+G than in NC-G, generating an OR of 104 (95% CI :11,4-136,6; p <0.0001). The results of this study show the association of rs1143634 polymorphisms in the IL1-? gene, rs1800796 in the IL6 gene, and rs1800629 in the TNF-? gene with genetic susceptibility to PD and/or DM development in the adult population studied, and association of the polymorphism rs1800796 in the IL6 gene with genetic susceptibility for PD and/or DM development in the children and adolescents population studied / Mestrado / Estomatologia / Mestra em Estomatopatologia

The molecular mechanisms underlying susceptibility to severe respiratory syncytial virus (RSV) infection remain poorly understood. Herein, we report on the role of osteopontin (OPN) in regulation of RSV infection in human epithelial cells and how interleukin-1 beta (IL-1β), a cytokine secreted soon after RSV infection, when persistently expressed can induce OPN expression leading to increased viral infection. We first compared OPN expression in two human epithelial cell lines: HEK-293 and HEp-2. In contrast to HEp-2, HEK-293 expresses low levels of pro-caspase-1 resulting in decreased IL-1β expression in response to RSV infection. We found a correlation between low IL-1β levels and a delay in induction of OPN expression in RSV-infected HEK-293 cells compared to HEp-2. This phenomenon could partially explain the high susceptibility of HEp-2 cells to RSV infection versus the moderate susceptibility of HEK-293 cells. Also, HEK-293 cells expressing low levels of pro-caspase-1 exhibit decreased IL-1β expression and delayed OPN expression in response to RSV infection. HEK-293 cells incubated with human rIL-1β showed a dose-dependent increase in OPN expression upon RSV infection. Also, incubation with rOPN increased RSV viral load. Moreover, HEp-2 cells or mice infected with a mucogenic RSV strain RSV-L19F showed elevated levels of OPN in contrast to mice infected with the laboratory RSV strain rA2. This correlated with elevated levels of OPN following infection with RSV-L19F compared to rA2. Together, these results demonstrate that increased OPN expression is regulated in part by IL-1β, and the interplay between IL-1β and OPN signaling has a pivotal role in the spread of RSV infection.

Inflammation is a biological defence mechanism applied by living organisms against foreign invaders. In the response to DAMPs and PAMPs, organisms use inflammatory multi-protein complexes to fight the attackers. The most studied inflammasome proteins are NLRP3, ASC and Caspase-1. This study is aimed at understanding the role of NEK7 protein in the NLRP3 inflammasome’s activation, using CRISPR/Cas9 system. To determine the effect of CRISPR/Cas9 and transfection, mRNA expression was analyzed. The results obtained suggest that neither the transfection nor the NEK7 protein knockout have sufficiently worked. This study could not experimentally establish that NEK7 triggers NLRP3 inflammasome activation because ELISA was not conducted to verify the levels of cytokines emitted, due to there being no statistical differences between the samples. Above all, the research question in this thesis project was not answered because the instability of the ACTB reference gene negatively influenced the results. However, previous related studies conclude that NEK7 plays a crucial role in the activation of the NLRP3 inflammasome.

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Indiana University-Purdue University Indianapolis (IUPUI) / Recent studies in anesthetized rats suggest that autonomic effects relating to thermoregulation that are evoked from the preoptic area (POA) may be mediated through activation of neurons in the dorsomedial hypothalamus (DMH). Disinhibition of neurons in the DMH produces not only cardiovascular changes but also increases in plasma adrenocorticotropic hormone (ACTH) and locomotor activity mimicking those evoked by microinjection of muscimol, a GABAA receptor agonist and neuronal inhibitor, into the POA. Therefore, I tested the hypothesis that all of these effects evoked from the POA are mediated through neurons in the DMH by assessing the effect of bilateral microinjection of muscimol into the DMH on the changes evoked by microinjection of muscimol into the POA in conscious rats. In addition, I tested the hypothesis that neurons in the DMH mediate a specific response that is thought to signal through the POA, the activation of the HPA axis evoked by systemic administration of the inflammatory cytokine IL-1β. After injection of vehicle into the DMH, injection of muscimol into the POA elicited marked increases in heart rate, arterial pressure, body temperature, plasma ACTH and locomotor activity and also increased Fos expression in the hypothalamic paraventricular nucleus (PVN), a region known to control the release of ACTH from the adenohypophysis, and the raphe pallidus, a medullary region known to mediate POA-evoked sympathetic responses. Prior microinjection of muscimol into the DMH produced a modest depression of baseline heart rate, arterial pressure, and body temperature but completely abolished all changes evoked from the POA. Microinjection of muscimol just anterior to the DMH had no effect on POA-evoked autonomic and neuroendocrine changes. Inhibition of neuronal activity in the DMH only partially attenuated the increased activity of the HPA axis following systemic injections of IL-1β. Thus, neurons in the DMH mediate a diverse array of physiological and behavioral responses elicited from the POA, suggesting that the POA represents an important source of inhibitory tone to key neurons in the DMH. However, it is clear that the inflammatory cytokine IL-1β must employ other pathways that are DMH-, and possibly POA-, independent to activate the HPA axis.