O uso de álcool está aumentando em nossa sociedade e permanece associado a inúmeros problemas sociais, econômicos e de saúde. De fato, o álcool e os problemas de saúde associados a ele exercem um impacto importante na prática médica e representam um dos maiores desafios da saúde pública. O consumo de álcool na sociedade contemporânea é geralmente aceito positivamente, dificultando o reconhecimento de certos padrões de consumo como doença. O alcoolismo é um transtorno recidivante crônico caracterizado pela ingestão compulsiva de quantidades excessivas de etanol, perda de controle em sua ingestão, comportamento inadequado e a presença de um estado emocional negativo. O consumo de quantidades nocivas de álcool resulta em danos físicos e psicológicos e o vicio é um transtorno psiquiátrico que afeta as funções executivas, causando perda de interesse em outros alvos do prazer e comportamento compulsivo de busca por drogas. O álcool interage com vários sistemas neurológicos. O presente trabalho analisou a resposta imunoinflamatória no tecido cerebral de camundongos CRAMP knockout (KO) jovens e tipo selvagem (WT) submetidos ao modelo de intoxicação alcoólica, com o objetivo de investigar o impacto de CRAMP na dependência alcoólica no adolescente. O CRAMP é um peptídeo antimicrobiano com efeitos pleotrópicos e, até onde sabemos, seu papel nunca foi investigado nesse sentido. Também analisamos a secreção de vários neuropeptídeos, proteínas e citocinas. Nossos resultados mostraram uma diferença significativa na ingestão de etanol entre os animais comparados CRAMP KO e WT, o que foi relacionado a um aumento nos níveis cerebelares de IL-1beta. Concluimos que os pepitídeos antimicrobianos podem ter um papel importante na resposta imunoinflamatória que ocorre durante o etilismo agudo / The use of alcohol is increasing in our society and remains associated with countless social, economic and health problems. In fact, alcohol and the health issues associated to its abuse exert an important impact on medical practice and represent one of the biggest challenges of public health. The consumption of alcohol in contemporary society is generally accepted positively, making certain patterns of consumption very difficult to be recognized as a disease. Alcoholism is a chronic relapsing disorder characterized by compulsive ingestion of excessive amounts of ethanol, loss of control in its intake, inappropriated behavior and the presence of a negative emotional state. The consumption of harmful amounts of alcohol results in physical and or psychological damage and addiction is a psychiatric disorder that affects the executive functions, causing loss of interest in other aspects of life and a compulsive behavior. Alcohol interacts with several neurologic systems. The present work analyzed the immuno-inflammatory response in the brain tissue of young CRAMP knockout (KO) and wild-type (WT) mice submitted to a model of alcohol intoxication, in order to investigate the impact of CRAMP in teenager alcohol addiction. CRAMP is an antimicrobial peptide with pleotropic effects and, as far as we know, its role had never been investigation in this regard. We also analysed the secretion of several neuropeptides, proteins and cytokines. Our results showed a significant difference in ethanol intake when CRAMP KO and WT animals were compared, which was related to an increase in the cerebellar levels of IL-1beta. We conclude that antimicrobial peptides may play an important role in the immunoinflammatory response that occurs during acute alcoholism

The effect of interleukin-1 beta (IL-1β) on the electrical properties of sensory neurons was assessed at comparable levels and exposure times to those found in animal models of neuropathic pain. Experiments involved whole cell current- or voltage-clamp recordings from rat dorsal root ganglion (DRG) neurons in defined medium, neuron enriched cultures. 5-6 days exposure to 100 pM IL-1β produced neuron specific effects. These included an increase in the excitability of medium diameter and small diameter isolectin B4 (IB4)-positive neurons that was comparable to that found after peripheral nerve injury. By contrast, a reduction in excitability was observed in large diameter neurons, while no effect was found in small diameter IB4-negative neurons. Further characterization of changes in medium and small IB4-positive neurons revealed that some, but not all, effects of IL-1β were mediated through its receptor, IL-1RI. Using appropriate voltage protocols and/or ion substitutions, it was found that neuron specific changes in several ionic currents, including alterations in hyperpolarization activated inward current (IH) and decreases in various K+ currents contribute to the increased excitability produced by IL-1β. Overall, these studies revealed that: 1. The effects of long-term exposure of DRG neurons to IL-1β are reflective of the enduring increase in primary afferent excitability reported after peripheral nerve injury. This expands the recognized role of IL-1β in acute inflammatory pain to neuropathic pain. 2. Hyperexcitability in medium neurons exposed to IL-1β likely includes mixed populations of neurons corresponding to nociceptive and non-nociceptive primary afferent fibres and, therefore, has relevance to hyperalgesia and allodynia, respectively. 3. The responsiveness of small IB4-positive neurons, but not IB4-negative, to prolonged IL-1β exposure is consistent with the suggestion that small IB4-negative afferents are involved in inflammatory pain, while small IB4-positive afferents are involved neuropathic pain. 4. The identification of receptor mediated effects and several contributing ionic mechanisms, may have relevance to the development of new therapeutic approaches to neuropathic pain. 5. IL-1β can contribute to increased neuronal excitability by mechanisms that are independent of IL-1RI signalling. This should be taken into account when targeting IL-1β, or more specifically IL-1RI, in the management of neuropathic pain.

Adipositas und ihre Folgeerkrankungen stellen eine wachsende medizinische Herausforde- rung globalen Ausmaßes dar. Im Rahmen der Adipositasforschung wurde das Fettgewebe als endokrines Organ identifiziert. Von ihm sezernierte Proteine, die sogenannten Adipo- kine, beeinflussen maßgeblich Insulinresistenz und Gefäßverletzbarkeit. Adipositas geht im Fettgewebe mit einer subklinischen chronischen Entzündung einher, die zu einer erhöhten Sekretion von proinflammatorischen Adipokinen führt. Die verstärkte Anwesenheit dieser Proteine ist mit den Komplikationen der Adipositas assoziiert. Die vorliegende Arbeit be- fasst sich mit dem Einfluss von Interleukin (IL)-1β, einem wichtigen Entzündungsmediator des Organismus, auf die Sekretion der proinflammatorischen Adipokine tissue inhibitor of metalloproteinase (TIMP)-1, serum amyloid A (SAA)-3, Lipocalin-2 und Chemerin. Die zugrundeliegenden Untersuchungen wurden mit 3T3-L1- und braunen Adipozyten durch- geführt. Es erfolgte der Nachweis auf mRNA- sowie auf Proteinebene. Der Einsatz von spe- zifischen Inhibitoren erlaubte den Rückschluss auf grundlegende Signalwege. Für alle vier untersuchten Adipokine konnte eine signifikante dosis- und zeitabhängige Steige- rung der mRNA- und Proteinexpression durch IL-1β nachgewiesen werden. Die Transduktion des IL-1β-Signals erfolgte im Falle von Lipocalin-2 und SAA-3 über nuclear factor (NF)-κB und janus kinase (Jak)-2, bei TIMP-1 lediglich über Jak-2 und in Bezug auf Chemerin über NFκB, Jak-2, p44/42 mitogen-activated protein kinase und Phosphatidylinositol-3-Kinase. Die in dieser Arbeit nachgewiesenen Expressions- und Sekretionssteigerungen von TIMP-1, SAA-3, Lipocalin-2 und Chemerin in braunen und weißen Adipozyten festigen IL-1β als einen entscheidenden Mediator proinflammatorischer Prozesse im Fettgewebe. Eine umfassende Bewertung der Funktion von IL-1β im Fettgewebe, insbesondere im Zustand der Adipositas, muss jedoch in weitergehenden Studien erfolgen.

No description available.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

Thesis (Ph.D. in Clinical Science) -- University of Colorado Denver, 2008. / Typescript. Includes bibliographical references (leaves 96-120). Free to UCD Anschutz Medical Campus. Online version available via ProQuest Digital Dissertations;

Introduction: Preeclampsia (PE) is characterized by the occurrence of gestational hypertension and proteinuria. Cytokines as IL-1 can cause an exaggerated inflammatory process causing oxidative stress and endothelial damage. In PE, and considering the ethnic variation, these cytokines are encoded through their encoding gene: among them the IL1B gene. Objectives: In order to investigate the association of polymorphisms in the IL1B gene in women with preeclampsia and the influence of possible maternal, environmental and genetic risk factors, a case-control study was conducted in two reference maternity hospitals in the state of Sergipe. Methods: An interview was done and the DNA of 456 volunteers collected for analysis. The predominance of melanoderma and feoderma individuals is emphasized here: few studies with this population are found in the literature regarding IL1B and PE. The genetic analysis aimed the representation of the gene integrally and for this purpose four tagSNPs for IL1B gene were analyzed: rs1143643, rs1143633, rs1143634 and rs1143630. The analysis of the association between the haplotypes and p values were calculated using the THESIAS software. Chi square test, Fisher test, Odds Ratio, confidence interval and multivariate logistic regression were performed. Results: Factors such as rs1143630 polimorphisms, obesity, primiparity, cesarean delivery and low level of consciousness at the time of admission to the health service were related to PE. Suggestive evidence of association between "T" allele in rs1143630 and preeclamptic women was observed. Conclusions: Clinical factors and IL1B polimorphisms were associated with severe preeclampsia. In many aspects, avoid maternal deaths related with preeclampsia is one of the most important and urgent challenges of obstetrics today. / Introdução: Pré-eclâmpsia (PE) é uma doença caracterizada pela ocorrência de hipertensão e proteinúria na gestação. Algumas citocinas como a IL-1 podem causar um processo inflamatório exagerado, causando lesão endotelial e estresse oxidativo. Na PE, e considerando-se a variação étnica, citocinas são codificadas através de seu gene codificador, entre eles o gene IL1B. Objetivos: Com o objetivo de investigar a associação entre polimorfismos no gene IL1B em mulheres com PE e a influência de possíveis fatores de risco maternos e ambientais, realizou-se um estudo do tipo caso-controle em duas maternidades de referência no estado de Sergipe. Métodos: Uma entrevista foi realizada e o DNA de 456 voluntárias foi coletado para análise. Ressalta-se aqui a predominância de indivíduos melanodermas e feodermas: poucos estudos com esta população são encontrados na literatura utilizando essa população em particular relacionando o gene IL1B e PE. A análise genética objetivou a representação do gene integralmente e para tanto quatro tagSNPs do gene IL1B foram analisados, sendo eles: rs1143643, rs1143633, rs1143634 e rs1143630. A análise da associação entre os haplótipos e valores de P foram calculados usando o software THESIAS. Teste de qui-quadrado, Teste de Fisher, Odds Ratio, intervalo de confiança e regressão logística foram realizados. Resultados: Fatores como polimorfismos do rs1143630, obesidade, primiparidade, parto cesáreo e baixo nível de consciência no momento da admissão no serviço de saúde relacionaram-se à PE. Foi identificada evidência sugestiva da associação do alelo “T” no rs1143630 no grupo de mulheres com PE. Conclusão: Fatores clínicos e polimorfismos do gene IL1B estiveram associados à ocorrência de pré-eclâmpsia. Em muitos aspectos, combater morte materna causada por pré-eclâmpsia é um dos desafios mais importantes e urgentes da obstetrícia na atualidade.

Les inflammasomes sont des complexes protéiques intracellulaires qui ont un rôle majeur dans l'immunité innée. Leur activation conduit à la mort de la cellule dans un contexte hyperinflammatoire. Compte-tenu des effets potentiellement délétères, tissulaires et systémiques, les inflammasomes sont strictement régulés. A l'heure actuelle, la compréhension des mécanismes conduisant à leur activation et leur régulation reste partielle. Dans une première partie de cette thèse, nous avons utilisé une technique de biotinylation proximale (BioID) pour identifier les protéines interagissant avec l'inflammasome. Nous avons identifié 111 protéines dont la relation étroite avec l'inflammasome était vraisemblable. Parmi ces 111 protéines, 25% avaient d'ailleurs déjà été décrites comme des protéines interagissant avec le complexe. L'identification d'un adaptateur majeur de l'autophagie, p62/sequestosome-1 (p62), nous a conduit à focaliser notre attention sur son rôle dans la régulation de l'inflammasome. Nous avons d'abord démontré que l'interaction entre p62 et l'inflammasome existait, sur le plan biochimique. Par la suite, nous avons prouvé que p62 était un substrat du complexe et que l'activation de ce dernier entrainait le clivage de p62 au niveau d'un résidu aspartique en position 329. Enfin, nous avons caractérisé les conséquences fonctionnelles de ce clivage, en montrant que les fragments protéiques générés entrainaient une régulation positive ou négative du complexe. Nous avons alors émis l'hypothèse que p62 pourrait réguler l'inflammasome de manière différente selon le signal activateur. Dans une seconde partie, translationnelle, nous nous sommes intéressés aux conséquences des mutations dans la séquence de gènes codant les constituants de l'inflammasome ou des protéines régulatrices du complexe. Celles-ci sont à l'origine des maladies auto-inflammatoires monogéniques. Ces maladies sont caractérisées par des épisodes récurrents de fièvre associés variablement à d'autres symptômes systémiques. La plus fréquente est la fièvre méditerranéenne familiale (FMF), avec une prévalence estimée entre 1 et 5 pour 10 000 habitants en France. Les mutations du gène MEFV, codant la pyrine, sont à l'origine de la FMF. La pyrine peut induire la formation d'un inflammasome spécifique. Récemment, le mécanisme d'activation de l'inflammasome pyrine a été mieux caractérisé : certaines toxines (comme la toxine B du Clostridium difficile, TcdB) induisent l'activation de l'inflammasome pyrine. Nous avons utilisé ces nouvelles connaissances afin d'explorer les conséquences de l'activation de l'inflammasome pyrine par la TcdB dans les monocytes des patients atteints de FMF, comparés aux monocytes de donneurs sains. Nos résultats indiquent que ces mutations induisent un abaissement du seuil d'activation de l'inflammasome pyrine. Par ailleurs, les corrélations génotype/phénotype indiquent qu'il existe un effet de dosage génétique, en lien avec le nombre d'allèles mutés. Ces résultats ouvrent de nouvelles perspectives pour les patients atteints de FMF, tant dans la compréhension de la physiopathologie que dans la possibilité de mise au point de tests fonctionnels pour le diagnostic de la maladie / Inflammasomes are intracellular multiprotein complexes that have a major role in innate immunity. Their activation leads to hyperinflammatory cell death. In view of potentially deleterious effects, the inflammasomes are strictly regulated. At present, the understanding of the mechanisms leading to their activation and regulation remains partial. In a first part of this thesis, we used a technique of proximity-dependent biotinylation (BioID) to identify the proteins interacting with the inflammasome. We identified 111 proteins with a close relationship to the inflammasome. Among these 111 proteins, 25% had already been described as proteins interacting with the complex. The identification of a major adaptor of autophagy, p62/sequestosome-1 (p62), led us to focus our attention on its role in the regulation of inflammasome. We first demonstrated that the interaction between p62 and inflammasome was real, at the biochemical level. Subsequently, we proved that p62 was a substrate of the complex and that the activation of the latter led to the cleavage of p62 at the aspartate 329. Finally, we characterized the functional consequences of this cleavage and showed that the protein fragments generated led to a positive or negative regulation of the complex. We thus hypothesized that p62 could regulate the inflammasome differently according to the activator signal. In a second translational part, we looked at the consequences of mutations in the sequence of genes coding components of the inflammasome or proteins regulating it. These are the cause of monogenic auto-inflammatory diseases. These diseases are characterized by recurrent episodes of fever associated with other systemic symptoms. The most frequent is Familial Mediterranean Fever (FMF), with prevalence estimated at between 1 and 5 per 10 000 inhabitants, in France. Mutations of the MEFV gene, encoding pyrin, cause FMF. Pyrine may trigger the formation of a specific inflammasome. Recently, the mechanism of activation of the pyrin inflammasome has been better characterized: toxins (such as toxin B of Clostridium difficile, TcdB) induce the activation of the pyrin inflammasome. We used this new knowledge to investigate the consequences of TcdB on pyrin inflammasome activation in monocytes from FMF patients compared to monocytes from healthy donors. Our results indicate that these mutations induce a decreased threshold of activation of the pyrin inflammasome. In addition, genotype / phenotype correlations indicate a gene-dosage effect, related to the number of mutated alleles. These results open new perspectives for patients with FMF, in understanding the pathophysiology of the diseass and in developing functional diagnostic tests

Embora já tenha sido comprovada a ação adjuvante da SBA-15, pouco se sabe sobre o seu mecanismo molecular que leva a modulação positiva da resposta imunológica. Foi avaliada a ativação do inflamassoma NLRP3, sobre estímulos de SBA-15, em macrófagos de camundongos C57BL/6. Como parâmetro dessa ativação, foi analisada a produção de IL-1β por ELISA. A SBA-15 foi capaz de induzir a produção de IL-1β a níveis semelhantes quando comparado com um agonista de NLRP3 (Nano-SiO2), sugerindo a ativação do inflamassoma. Para avaliar o envolvimento da caspase-1, nos resultados obtidos com a SBA-15, os macrófagos foram estimulados com sílica na presença do inibidor de caspase-1, e como esperado, a produção de IL-1β foi restaurada para o seu nível basal. A ativação do inflamassoma, por estímulos da SBA-15, parece ser parcialmente dependente da fagocitose e da produção das espécies reativas do oxigênio. Além disso, foi visto que a SBA-15 não induz a produção de IL-6, confirmando que essa sílica está envolvida na via do inflamassoma e não em outras vias, como por exemplo, NF-κB. / Although it has already been proven adjuvant action of SBA-15, little is known about its molecular mechanism leading to positive modulation of the immune response. NLRP3 inflammasome activation was evaluated on SBA-15 stimulation in C57BL/6 mice macrophages. As this parameter activation, it analyzed the production of IL-1β by ELISA. The SBA-15 was able to induce the production of IL-1β at levels similar when compared to an agonist of NLRP3 (Nano-SiO2), suggesting the activation of the inflammasome. To assess the involvement of caspase-1, the results obtained with SBA-15, the macrophages were stimulated with silica in the presence of caspase-1 inhibitor, and as expected, IL-1β production was restored to its baseline level. Activation of the inflammasome, by stimuli of SBA-15, appears to be partly dependent on phagocytosis and production of reactive oxygen species. In addition, it was seen that the SBA-15 does not induce IL-6 production, confirming that silica is involved inflammasome the path of and not in other ways, eg, NF-κB.

Introduction: Preeclampsia (PE) is characterized by the occurrence of gestational hypertension and proteinuria. Cytokines as IL-1 can cause an exaggerated inflammatory process causing oxidative stress and endothelial damage. In PE, and considering the ethnic variation, these cytokines are encoded through their encoding gene: among them the IL1B gene. Objectives: In order to investigate the association of polymorphisms in the IL1B gene in women with preeclampsia and the influence of possible maternal, environmental and genetic risk factors, a case-control study was conducted in two reference maternity hospitals in the state of Sergipe. Methods: An interview was done and the DNA of 456 volunteers collected for analysis. The predominance of melanoderma and feoderma individuals is emphasized here: few studies with this population are found in the literature regarding IL1B and PE. The genetic analysis aimed the representation of the gene integrally and for this purpose four tagSNPs for IL1B gene were analyzed: rs1143643, rs1143633, rs1143634 and rs1143630. The analysis of the association between the haplotypes and p values were calculated using the THESIAS software. Chi square test, Fisher test, Odds Ratio, confidence interval and multivariate logistic regression were performed. Results: Factors such as rs1143630 polimorphisms, obesity, primiparity, cesarean delivery and low level of consciousness at the time of admission to the health service were related to PE. Suggestive evidence of association between "T" allele in rs1143630 and preeclamptic women was observed. Conclusions: Clinical factors and IL1B polimorphisms were associated with severe preeclampsia. In many aspects, avoid maternal deaths related with preeclampsia is one of the most important and urgent challenges of obstetrics today. / Introdução: Pré-eclâmpsia (PE) é uma doença caracterizada pela ocorrência de hipertensão e proteinúria na gestação. Algumas citocinas como a IL-1 podem causar um processo inflamatório exagerado, causando lesão endotelial e estresse oxidativo. Na PE, e considerando-se a variação étnica, citocinas são codificadas através de seu gene codificador, entre eles o gene IL1B. Objetivos: Com o objetivo de investigar a associação entre polimorfismos no gene IL1B em mulheres com PE e a influência de possíveis fatores de risco maternos e ambientais, realizou-se um estudo do tipo caso-controle em duas maternidades de referência no estado de Sergipe. Métodos: Uma entrevista foi realizada e o DNA de 456 voluntárias foi coletado para análise. Ressalta-se aqui a predominância de indivíduos melanodermas e feodermas: poucos estudos com esta população são encontrados na literatura utilizando essa população em particular relacionando o gene IL1B e PE. A análise genética objetivou a representação do gene integralmente e para tanto quatro tagSNPs do gene IL1B foram analisados, sendo eles: rs1143643, rs1143633, rs1143634 e rs1143630. A análise da associação entre os haplótipos e valores de P foram calculados usando o software THESIAS. Teste de qui-quadrado, Teste de Fisher, Odds Ratio, intervalo de confiança e regressão logística foram realizados. Resultados: Fatores como polimorfismos do rs1143630, obesidade, primiparidade, parto cesáreo e baixo nível de consciência no momento da admissão no serviço de saúde relacionaram-se à PE. Foi identificada evidência sugestiva da associação do alelo “T” no rs1143630 no grupo de mulheres com PE. Conclusão: Fatores clínicos e polimorfismos do gene IL1B estiveram associados à ocorrência de pré-eclâmpsia. Em muitos aspectos, combater morte materna causada por pré-eclâmpsia é um dos desafios mais importantes e urgentes da obstetrícia na atualidade.