Due to the wide acceptance of the Word Wide Web Consortium (W3C) XPath language specification, native indexing for XML is needed to support path expression queries efficiently. XPath describes the different document tree relationships that may be queried as a set of axes. Many recent proposals for XML indexing focus on accelerating only a small subset of expressions possible using these axes. In particular, queries by ordinal position and updates that alter document structure are not well supported. A more general indexing solution is needed that not only offers efficient evaluation of all of the XPath axes, but also allows for efficient document update. We introduce MASS, a Multiple Axis Storage Structure, to meet the performance challenge posed by the XPath language. MASS is a storage and indexing solution for large XML documents that eliminates the need for external secondary storage. It is designed around the XPath language, providing efficient interfaces for evaluating all XPath axes. The clustered organization of MASS allows several different axes to be evaluated using the same index structure. The clustering, in conjunction with an internal compression mechanism exploiting specific XML characteristics, keep the size of the structure small which further aids efficiency. MASS introduces a versatile scheme for representing document node relationships that always allows for efficient updates. Finally, the integration of a ranked B+ tree allows MASS to efficiently evaluate XPath axes in large documents. We have implemented MASS in C++ and measured the performance of many different XPath expressions and document updates. Our experimental evaluation illustrates that MASS exhibits excellent performance characteristics for both queries and updates and scales well to large documents, making it a practical solution for XML storage. In conjunction with text indexing, MASS provides a complete solution from XML indexing.

Introdução: A neurobiologia dos transtornos de ansiedade (TA) é complexa e envolve interações ambientais e genéticas ainda não conhecidas. Esses transtornos, comumente, iniciam durante a infância e adolescência, persistindo ao longo da vida. O comprometimento da resposta biológica frente ao estímulo estressor, encontrado em muitos pacientes com TA, sugere a influência do eixo hipotálamo-hipófise-adrenal (HHA) nestes transtornos e, portanto, os polimorfismos associados ao eixo HHA poderiam ser estudados em genes candidatos. Os estudos que almejam entender a etiologia dos TA devem, também, explorar as alterações epigenéticas (incluindo a metilação do DNA) decorrentes das influências ambientais. Objetivos: Estudar, em adolescentes, polimorfismos genéticos funcionais do eixo HHA, interações Gene x Ambiente (G x A) e metiloma do DNA, considerando as diferentes trajetórias dos TA. Métodos: Foi realizada a extração de DNA das células do epitélio bucal de 228 adolescentes (131 casos e 97 controles para os TA) e foram genotipados, por PCR em tempo real, polimorfismos funcionais envolvidos com o eixo HHA (FKBP5: rs3800373, rs9296158, 3800373, rs9296158, 3800373, rs9296158, rs1360780, rs9470080 rs1360780, rs9470080 e rs4713916; NR3C1NR3C1 : rs6198;: rs6198;: rs6198; NR3C2NR3C2 : rs2070951;: rs2070951;: rs2070951; CRHR1CRHR1 CRHR1 : rs878886 : rs878886 e SERPINA6 SERPINA6 : rs746530) : rs746530) . Os participantes responderam à escala auto-aplicativa SCARED (Screen for Children Anxiety Related Emotional Disorder – Children rated) e realizaram entrevistas semiestruturadas para avaliação diagnóstica utilizando o K-SADS-PL (Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime). O questionário CTQ (Childhood Trauma Questionnaire) foi aplicado em 90 adolescentes (54 casos e 36 controles para os TA) para avaliar a interação entre o trauma emocional e o polimorfismo do gene NR3C2 nos níveis séricos de BDNF. Uma subamostra de adolescentes (n=47) foi reavaliada, cinco anos após a primeira coleta, através das mesmas entrevistas psiquiátricas e nova extração de DNA salivar. Alguns participantes, na última avaliação, responderam ao MINI (Mini International Neuropsychiatric Interview) apropriado para a idade atual. A amostra foi organizada em 4 grupos, conforme o diagnóstico dos TA e o ano da coleta de saliva (anos de 2008 e 2013): (1) Desenvolvimento típico da adolescência (Controle; n=14); (2) Incidentes para os TA (ITA; n=11); (3) Persistentes para os TA (Caso; n=14) e (4) Remitentes para os TA (RTA; n=08). O metiloma do DNA foi analisado com o Infinium HumanMethylation 450 BeadChip da Illumina. Resultados: Não foi encontrada associação entre os polimorfismos estudados e os TA. Em relação à interação G x A, sugere-se que o polimorfismo rs2070951 do gene NR3C2 modera a associação entre negligência física e os níveis séricos de BDNF. Do ponto de vista epigenético, foi observada, nos grupos ITA e RTA, vias biológicas com padrão homogêneo e relacionadas ao sistema nervoso. Já nos grupos casos e controles para os TA, foram evidenciadas vias biológicas com padrão mais heterogêneo. Um perfil de hipometilação do DNA foi predominante nas vias encontradas. Na análise transversal, nós encontramos padrões opostos de metilação do DNA, conforme o período desenvolvimental avaliado: hipometilação no início da adolescência e hipermetilação em jovens adultos. Conclusão: Esse estudo abordou, em uma amostra de adolescentes, aspectos genéticos (genes candidatos envolvidos com o eixo HHA), ambientais (trauma emocional) e epigenéticos (metiloma do DNA) dos TA. Os achados sugerem que, embora sem associações entre os TA e genes envolvidos no eixo HHA, existe uma interação entre a presença de trauma emocional, polimorfismo genético do eixo HHA e marcadores biológicos. Os achados do metiloma do DNA sugerem, também, influências epigenéticas no curso dos TA. Novos estudos devem ser delineados para corroborar as influências genéticas e ambientais neste transtorno. / Background: The neurobiology of Anxiety Disorders (AD) is complex and involves environmental and genetic interactions understood. These disorders may have their onset during childhood and adolescence, persisting throughout life. The impairment of biological response against the stressor stimulus, described in many patients with AD, suggests a possible role of genetic polymorphisms of the hypothalamic-pituitary-adrenal (HPA) axis in these individuals. Studies that aim to understand the etiology of AD should also explore the epigenetic changes (including DNA methylation) arising from environmental influences. Objective: To study, in adolescents, functional genetic polymorphisms of HPA axis, Gene x Environment (G x E) interactions and DNA methylome, considering different AD outcomes. Methods: Saliva DNA was extracted from 228 adolescents (131 cases and 97 controls to AD) and we genotyped, by real time PCR, the functional polymorphisms involved with HPA axis (FKBP5: rs3800373, rs9296158, rs1360780, rs9470080 and rs4713916; 3800373, rs9296158, rs1360780, rs9470080 and rs4713916; 3800373, rs9296158, rs1360780, rs9470080 and rs4713916; 3800373, rs9296158, rs1360780, rs9470080 and rs4713916; 3800373, rs9296158, rs1360780, rs9470080 and rs4713916; 3800373, rs9296158, rs1360780, rs9470080 and rs4713916; 3800373, rs9296158, rs1360780, rs9470080 and rs4713916; NR3C1NR3C1 : rs6198; : rs6198; : rs6198; NR3C2NR3C2 : rs2070951; : rs2070951; CRHR1CRHR1CRHR1 CRHR1: rs878886 and : rs878886 and : rs878886 and : rs878886 and SERPINA6 SERPINA6 SERPINA6SERPINA6 : rs746530) : rs746530) . Participants responded to the scale self-applied Screen for Children Anxiety Related Emotional Disorder – Children rated (SCARED) and were diagnosed according to the Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime (K-SADS-PL). The Childhood Trauma Questionnaire (CTQ) was applied in 90 adolescents (54 cases e 36 controls to AD) to evaluate the interaction between emotional trauma and the NR3C2 polymorphism in the serum levels of BDNF. A sub-sample of adolescents (n = 47) was reassessed five years after the first evaluation by the same psychiatric semi-structured interviews and new extraction salivary DNA was performed. Some participants in the last evaluation responded to Mini International Neuropsychiatric Interview (MINI), which is a semi structure interview appropriate for the present age. The sample was organized in four groups according to the diagnosis of AD and the year of saliva collection (2008 and 2013): (1) Typically Developing Controls (TDC; n = 14); (2) Incident Anxiety Disorder (IAD; n = 11); (3) Persistent Anxiety Disorder (PAD; n = 14); (4) Remittent Anxiety Disorder (RAD; n = 8). DNA methylome was evaluated with Infinium HumanMethylation450 BeadChip. Results: We did not find any association between the genetic polymorphisms and AD. Considering the G x E interaction we suggest that rs2070951 polymorphism of NR3C2 gene moderates the association between physical neglect and serum BDNF levels. When we evaluated the DNA methylome, we observed more homogeneous biological pathways and mostly related with nervous system in individuals from IAD and RAD groups. On the other hand, in the TDC and PAD groups, we found biological pathways with a more heterogeneous pattern. A DNA hypomethylation profile was found predominant in the pathways. In a cross-sectional analysis, we found opposite patterns of DNA methylation, as the developmental period assessed: hypomethylation at the beginning of adolescence and hypermethylation in young adults. Conclusion: This study addressed, in an adolescent sample, genetics (candidate genes linked to HPA axis), environmental (emotional trauma) and epigenetic (DNA methylome) aspects of AD. The findings suggest that although there are no associations between AD and genes involved in HPA axis, there is an interaction between the presence of trauma, genetic polymorphism involved in this axis and biomarkers. The DNA methylome findings also suggest epigenetic influences on the course of TA. Further studies should be designed to corroborate the genetic influences in this disorder.

La dépression majeure (DM) est une pathologie complexe et hétérogène associée avec des altérations du réseau cérébral, une dérégulation de l’axe hypothalamus-pituitaire-surrénales et avec des déficits de la neurogenèse adulte hippocampique (NHA). De nombreuses évidences pointent sur l’implication de la NHA dans les troubles de l’humeur et les troubles anxieux, ce qui a conduit à la formulation de l’ « hypothèse neurogénique », laquelle postule que des neurones néo-formés dans l’hippocampe du sujet adulte sont impliqués dans l’étiologie et dans l’efficacité du traitement de la DM. L’objectif de cette étude a été de déterminer le rôle des neurones formés à l’âge adulte après que les animaux aient été exposés à un stress ainsi que les mécanismes sous-jacents. Nos résultats suggèrent que l’augmentation de la neurogenèse est suffisante pour estomper les effets d’un stress chronique au niveau comportemental et hormonal, et donc pour induire un effet de type antidépresseur, comportementalement et physiologiquement. Les effets surviennent sans doute via le noyau du lit de la strie terminale antéro-dorsale. / Major depressive disorder (MDD) is a complex and heterogeneous disorder hypothesized to be associated with alterations in brain circuitry, dysregulations of the hypothalamic–pituitary–adrenal axis and impairments in adult hippocampal neurogenesis (AHN). Multiple lines of evidence point to the involvement of AHN in mood and anxiety disorders, leading to the formation of the “neurogenesis hypothesis”, which postulates that adult-born hippocampal neurons are involved in the etiology and treatment efficacy of MDD. The purpose of this study was to determine the role of adult-born neurons after the onset of stress exposure and the mechanism that underlies the observed results. Our results suggest that increasing neurogenesis is sufficient to buffer against the effects of chronic stress on certain behavioral and endocrine levels and thus to display antidepressant-like effects, both behaviorally and physiologically. Adult-born neurons might have exerted some of their effects via the anteromedial division of the bed nucleus of the stria terminalis (BSTMA).

Many studies have shown that short sleep duration and/or poor sleep quality is associated with increasing rates of cardiovascular (CVD) mortality and morbidity. One hypothesized explanation for this association has been that sleep loss is a type of chronic stress that induces dysregulation of biological systems that ultimately increase the risk of CVD. One biological system that has been thought to link sleep loss and CVD is the hypothalamus-pituitary-adrenal (HPA) axis. A number of studies using small or convenience samples have addressed the effects of sleep deprivation on cortisol. Only a few studies have examined the association of habitual short sleep duration and/or poor sleep quality with changes in the diurnal cortisol in population based-samples; those studies vary in their methodology and in findings. Another biological system that has been thought to link sleep loss and CVD is the autonomic nervous system (ANS), through overactivation of the sympathetic nervous system (SNS) and/or probably a withdrawal of the parasympathetic nervous system. Experimental studies have shown an association between the sleep stages and markers of the sympathetic system. However, very few studies of habitual sleep duration/sleep quality and ANS markers have been conducted. Even fewer studies have examined the association of habitual sleep duration and/or sleep quality and ANS responses to a stress challenge in a population-based sample. The findings again have been inconsistent probably due to the use of different methodology and different samples. This dissertation used measures of salivary diurnal cortisol as well as cortisol responses to a stress challenge protocol to assess the relationship of habitual sleep duration and/or sleep quality with diurnal cortisol profile in natural conditions and in response to a stress challenge protocol in a laboratory setting. Diurnal cortisol was assessed from up to 16 samples of salivary cortisol for two days. Cortisol responses to a stress challenge were assessed from four salivary samples taken during the stress challenge that was performed in a different day than the diurnal cortisol collection. To examine the relationship of habitual sleep duration and/or sleep quality and markers of the ANS, this dissertation used continuous cardiovascular measures (heart rate and heart rate variability) and four salivary amylase samples obtained during the stress challenge. The stress challenge included mental stress and orthostatic stress. Sleep duration and sleep efficiency (an objective measure of sleep quality) were assessed from 7-day actigraphy and sleep diaries. Insomnia symptoms (a subjective measure of sleep quality) were also assessed using a questionnaire that included the Women’s Health Initiative Insomnia rating scale (WHIIRS). We used mixed models so as to account for the repeated measures of diurnal salivary cortisol levels as well as the responses (reactivity and recovery) to the stress challenge tests. Chapter 1 presents an introduction to this dissertation discussing the relationship between short sleep duration and/or poor sleep quality and CVD morbidity and mortality. Chapter 2 presents a systematic literature review of studies of the association between habitual sleep duration and/or sleep efficiency and markers of neuro-endocrine systems: HPA and ANS. These are plausible mechanisms that link short and/or poor sleep to CVD morbidity and mortality. Chapter 3 presents our analyses of the relationship between short sleep duration and/or poor sleep quality and features of the diurnal cortisol. We hypothesized that those participants whose slept < 6 hours per night or whose sleep efficiency was < 85% would have higher cortisol levels on awakening, flatter cortisol awakening responses (CAR), and higher evening cortisol levels than participants who slept longer or slept better. We found that short sleepers had higher evening cortisol than the longer sleepers and that this association persisted after the adjustment for several known confounders. In chapter 4, we examined how the same groups of participants responded in terms of hormones (cortisol and amylase) and cardiovascular indices (heart rate (HR) and HR variability (HRV)) to a stress challenge test. We hypothesized that those participants who slept for a shorter time or whose sleep was of poorer quality would have more exaggerated responses to and less recovery from a stress challenge test than participants who slept longer or slept better. We found that participants with insomnia had exaggerated high frequency-HRV (HF-HRV) orthostatic reactivity. In an extended analysis, we found that participants who slept less than 7 hours/night had exaggerated heart rate reactivity to a mental stress test compared to participants who slept 7 or more hours/night, but this association was attenuated after adjustment for naps. Paradoxically, we also found that participants who slept less than 7 hours had higher HF-HRV recovery from mental stress compared to longer sleepers (≥7 hours). Short sleep duration or low sleep efficiency was not associated with cortisol or amylase responses to the stress challenge protocol. These findings suggest that sustained high evening cortisol levels and cardiovascular responses to a stress challenge may be among the mechanisms linking short/poor sleep and CV disease.

O presente estudo avaliou a relação dos parâmetros do eixo II dos Critérios de Diagnóstico de Pesquisa das Disfunções Temporomandibulares (RDC/TMD) na resposta terapêutica de pacientes com DTM. Trinta e três pacientes (média de idade de 33,2 anos ± 13,4) com artralgia da articulação temporomandibular (ATM) foram submetidos a três terapêuticas: laser de baixa potência (LBP) + piroxicam (LPi), LBP + placebo de piroxicam (L) e piroxicam + placebo de LBP (Pi). Os pacientes receberam a terapêutica por 10 dias. As avaliações foram feitas na 1a e 4a sessões de tratamento. A presença e intensidade de dor espontânea, dor à palpação e máxima abertura bucal foram mensuradas. A evolução destas foi comparada à classificação dos pacientes de acordo com os parâmetros do eixo II: grau de dor crônica (GDC), depressão e sintomas físicos não específicos (SFNE). Os dados foram analisados usando os testes de Fisher, Wilcoxon, t de Student, Kruskal-Wallis, U de Mann-Whitney e análise de variâncias, adotando-se p<0,05 como nível de significância. Foi possível observar melhora significativa (p<0,05) dos pacientes estudados quanto ao tempo em relação à EVA e dor à palpação tanto muscular quanto articular. Notou-se diferença estatística significante (p<0,05) entre os grupos de GDC e abertura bucal 4ª sessão e dor articular; entre os grupos de SFNE (incluindo e excluindo itens de dor) e palpação muscular 1ª sessão. Entre os grupos de GDC e as variáveis de EVA e palpação muscular foi encontrada diferença marginalmente significativa, apresentando p-valores próximos a 0,05 (p<0,1). Não foi encontrada diferença estatística significante entre os grupos de depressão e as variáveis consideradas. Portanto, foi observada relação entre alta incapacidade e maiores médias dor articular, grau de SFNE incluindo e excluindo itens de dor classificados como severos e palpação muscular inicial. A depressão não apresentou relação com as variáveis estudadas. / The aim of this study was assess the influence of Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) Axis II parameters on treatment outcome of low-level laser therapy (LLLT) associated with piroxicam in patients with TMD. Thirty-two patients (mean age 33.2 years old ± 13.4) with temporomandibular joint (TMJ) arthralgia were enrolled in the study and received three kinds of treatment: LLLT + piroxicam (LPi), LLLT + placebo piroxicam (L) e piroxicam + placebo LLLT (Pi). Patients were managed for ten days. Follow-up evaluations were done at the 1st and 4th consults. The presence and intensity of spontaneous pain through the visual analogue scale (VAS), painful palpation and mandibular maximum vertical opening were measured. The therapeutic outcomes of this population were compared between the classifications of axis II scores: graded chronic pain (GCP), depression and non-specific physical symptoms (NEPS). The data was analyzed using the following statistic models: Fisher\'s test, Student t test, Wilcoxon, Mann-Whitney U test, Kruskal-Wallis test and Analysis of Variance. The statistical significance level set was p<0.05. The variables VAS, painful joint and muscle palpation showed improvement over time. Significant difference was found between GCP groups and maximum mouth opening on the 4th session and painful TMJ palpation; between NEFS (including and excluding pain items) and painful muscle palpation on the 1st consult. Between groups of GCP and VAS\'s variables it was found a marginally significant relation, showing p-values near 0.05 (p<0.1). No statistical difference was found on the comparison between depression and the physical variables considered. Thus, it is possible to conclude that there is an association between high incapacity and higher mean values to painful joint palpation, severe NEPS including and excluding pain items and baseline painful muscle palpation. No relation between depression and the studied variables was observed.

Representações computacionais de formas podem ser criadas em ferramentas CAD ou geradas a partir de um objeto físico já existente. Esta última abordagem oferece como vantagens rapidez e fidelidade ao objeto original, que são os aspectos fundamentais em muitas aplicações, como Simulações Numéricas de Equações Diferenciais Parciais e Imagens Médicas. A reconstrução (ou geração de malhas superficiais) a partir de pontos amostrados de uma superfície de um objeto é um problema clássico de representação de formas. Nesta dissertação apresentamos um vasto levantamento bibliográfico deste tipo de reconstrução, classificando e descrevendo os principais trabalhos presentes na literatura. A partir do levantamento bibliográfico, selecionamos um conjunto de algoritmos sobre os quais foram realizadas comparações teóricas e empíricas cujos resultados são apresentados. Para finalizar, apresentamos aplicações de nossas implementações em Simulação Numérica de Equações Diferenciais Parciais e processamento de Imagens / Computational representations of shapes can be developed using CAD applications or created from data acquired from a real physical object. This latter is advantageous with respect to time and fidelity to the original object which are essential to several applications, such as Numerical Simulation of Partial Differential Equations and Medical Imaging. A classical shape representation problem is that of reconstruction (or superficial mesh generation) from points sampled over the surface of an object. In this Master\'s thesis we describe a broad survey of these reconstruction methods. We focus in the classification and characterization of the main algorithms proposed in the literature. From this survey, we selected some algorithms and conducted some theoretical and practical comparisons. We conclude this work describing applications of the algorithms implemented in Numerical Simulations of Differential Partial Equations and Image Processing

O ACTH é considerado o principal fator atuante no desenvolvimento, na manutenção e na esteroidogênese da glândula adrenal. No entanto, existem evidências que peptídeos N-Pomc, derivados do processamento da Proopiomelanocortina (POMC), possam atuar na manutenção do córtex adrenal, embora ainda não seja conhecida sua importância e mesmo quais são os peptídeos que atuam na suprarrenal. Nesse trabalho tivemos como objetivo avaliar os efeitos de peptídeos sintéticos de 28 aminoácidos derivados do N-Pomc (N-PomcCys, N-PomcMet e N-PomcSer) na morfologia e função da suprarrenal de camundongos, cujo gene Pomc foi condicionalmente silenciado com Tamoxifeno (Tmx). Foram obtidos animais machos adultos (CrePomcfloxflox) com um sistema \"knock-out\" condicional Cre-Lox induzível por Tmx, que foram tratados através de minibombas osmóticas por 21 dias com os peptídeos ou com salina, e como controle animais Pomcfloxflox não tratados. Foram analisados: 1) dados metabólicos; 2) a concentração de ACTH e de corticosterona plasmáticos; 3) a morfologia e a reconstrução anatômica da adrenal; 4) a capacidade funcional através da coloração com Oil Red O (ORO) e 5) a capacidade de proliferação através da expressão da proteína PCNA. A caracterização dos animais CrePomcfloxflox + Tmx após o silenciamento mostrou a redução de 60% da concentração de ACTH plasmático e esses animais apresentaram 1) redução do gasto energético, aumento da ingestão de alimentos e ganho de peso corpóreo; 2) alteração significante a área ou o volume das adrenais; 3) redução da produção de gotículas lipídicas e 4) redução do número de núcleos positivos para a proteína PCNA. Esses animais silenciados para a Pomc e tratados com os peptídeos N POMCCys e N-POMCMet apresentaram 1) aumento da corticosterona plasmática e apenas o N-POMCCys aumentou o ACTH plasmático; 2) aumento de núcleos marcados para PCNA. Concluímos, que os camundongos Cre Pomcflox/flox silenciados para a Pomc com o Tamoxifeno apresentaram alterações metabólicas, morfológicas e fisiológicas. A análise do efeito biológico dos peptídeos N-POMC mostrou ação desses peptídeos na função e na manutenção do córtex adrenal. / ACTH is considered the main active factor in the development, maintenance and steroidogenesis of the adrenal gland. However, there is evidence that N-Pomc peptides, derived from the processing of Proopiomelanocortina (POMC), may play a role in the maintenance of the adrenal cortex, although their importance is not yet known. In this work we aimed to evaluate the effects of synthetic peptides of 28 amino acids derived from N-Pomc (N-PomcCys, N-PomcMet e N-PomcSer) on the morphology and function of the adrenal cortex of mice whose Pomc gene was conditionally silenced with Tamoxifen (Tmx). Adult males (CrePomcfloxflox) were obtained with a Tmx inducible Cre-Lox conditional knock-out system, which were treated by osmotic minipumps for 21 days with the peptides or with saline, and as control untreated Pomcfloxfloxanimals. We analyzed: 1) metabolic data; 2) plasma ACTH and corticosterone concentration; 3) the morphology and the anatomical reconstruction of the adrenal; 4) functional capacity through staining with Oil Red O (ORO) and 5) the ability to proliferate through expression of PCNA protein. The characterization of the CrePomcfloxflox + Tmx animals after silencing showed a 60% reduction in plasma ACTH concentration and these animals presented 1) reduction of energy expenditure, increased food intake and body weight gain; 2) significant alteration of adrenal area or volume; 3) reduction of the production of lipid droplets and 4) reduction of the number of nuclei positive for the PCNA protein. These animals that were silenced to Pomc and treated with peptides N-PomcCys, N-PomcMet had 1) increase in plasma corticosterone and only N-POMCCys increased plasma ACTH; 2) increase of nuclei marked for PCNA. We conclude that CrePomcflox/flox mice silenced for Pomc with Tamoxifen presented metabolic, morphological and physiological alterations. The analysis of the biological effect of N-POMC peptides showed the action of these peptides on the function and maintenance of the adrenal cortex.

Daily rhythms in physiology and behaviour are orchestrated by theautonomously rhythmic cells of the suprachiasmatic nucleus (SCN).Restricting food intake to the rest phase of nocturnal rodents, leads to thedevelopment of meal anticipatory behaviour, corticosterone and bodytemperature. Given that lesions to the SCN fail to abolish meal anticipation, asecond oscillator of unknown location, referred to as the food-entrainableoscillator (FEO) is thought to exist. Although the site of the FEO is unknown,several hypothalamus nuclei, including the dorsomedial hypothalamus(DMH) are thought to play a role in meal anticipation. Given thatadrenalectomy is reported to abolish meal anticipation, an intact HPA axis isalso thought to contribute to the functioning of the FEO. Some forms ofobesity are characterised by high basal levels of circulating corticosterone. Inaddition, limited access to high fat diet, fails to induce the development ofrobust meal anticipation in rats. During our initial studies, the effect of a standard and 45% high fat diet onthe development of meal anticipatory behaviour and hypothalamic c-Fosexpression were investigated. Restricted access to high fat diet led toattenuation of meal anticipation compared to those fed standard diet. Thiswas concurrent with a failure to develop an anticipatory rise in DMH c-Fosexpression. A meal anticipatory rise in corticosterone is thought to benecessary for the presence of meal anticipation as well as adaptation ofmetabolism to daily restricted feeding. In the next set of studies, weconfirmed that restricted access to standard diet leads to the development ofa meal anticipatory rise in plasma corticosterone. In contrast we observed adramatic post-anticipatory rise in plasma corticosterone in rats givenrestricted access to the 45% high fat diet. We hypothesised that the highcorticosterone levels resulting from high fat diet were a contributing factor tothe lack of meal anticipation in high fat fed rats. With the aid of apharmacokinetic study, a suitable experiment was designed for daily dosingof a potent glucocorticoid receptor antagonist, RU486, with the aim ofrescuing meal anticipation in high fat fed rats. Interestingly, treatment withRU486 successfully rescued meal anticipation in high fat fed rats, butattenuated meal anticipation in standard diet restricted fed rats. In the finalseries of studies the effect of diet and feeding regime on glucose toleranceand metabolism were investigated. High fat feeding was found to reduceglucose tolerance in both ad lib and restricted fed rats, with RU486 treatmentimproving glucose tolerance in a time dependant manner. Restricted accessto food was found to induce post satiation lipogenesis in both standard dietfed and to a lesser extent in high fat fed rats, an effect which may bebeneficial in reducing obesity. Overall the results provide further insight intothe complex role of corticosterone in promoting or preventing mealanticipatory behaviour. An anticipatory rise in plasma corticosterone isrequired for meal anticipation, as repeated daily dosing of RU486 inhibit mealanticipation. The high basal levels of corticosterone in high fat fed rats mayprevent meal anticipation, insulin secretion and post-satiation lipogenesiswhich may in fact be a homeostatic mechanism to prevent obesity. Nonetheless, treatment with RU486 rescues behavioural meal anticipationand glucose tolerance.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A superativação do eixo ECA/AT1r está intimamente relacionada à síndrome metabólica e no organismo tem grande relação com o quadro de inflamação. A administração de frutose, seja por dieta ou pela água, tem sido usada como um modelo para a indução da superatividade desse eixo e para o estudo das vias inflamatórias relacionadas ao AT1r. Com isso, o objetivo deste trabalho foi avaliar se a administração de GW510156 poderia diminuir a superativação do eixo ECA/AT1r e consequentemente diminuir os danos causados pela dieta rica em frutose. Para isso foram utilizados camundongos machos C57Bl/6 que receberam uma dieta contendo 47% de frutose durante oito semanas ou uma dieta controle. Após oito semanas, os grupos foram redivididos aleatoriamente para o início da administração do GW501516 durante três semanas, totalizando quatro grupos experimentais. Os animais tratados apresentaram uma melhora da pressão arterial sistólica e também dos parâmetros urinários como proteinúria e ácido úrico. Houve ainda uma melhora dos triglicerídeo e ácido úrico plasmáticos. No tecido adiposo branco, o GW501516 foi capaz de diminuir a expressão dos componentes do eixo ECA/AT1r e também amenizou a inflamação causada pela dieta rica em frutose. No fígado, não houve alterações significativa do eixo, porém a fosforilação de JAK2 dependente de AT1r foi diminuída e consequentemente houve uma menor ativação das células estreladas no grupo que recebeu o GW501516. Além disso, as proteínas e genes relacionados à &#946;-oxidação foram aumentados com o tratamento e aqueles relacionados à lipogênese de novo, diminuídos o que resultou em menor esteatose no parênquima hepático. Os rins apresentaram uma melhora da inflamação induzida pelo eixo, apesar de o eixo também não ter apresentado diferenças significativas com o tratamento. Também não foram encontradas diferenças significativas na expressão proteica e gênica das proteínas antioxidantes. Com esses resultados podemos concluir que a curta administração do GW501516 pôde aliviar os efeitos inflamatórios e a esteatose hepática causada pela dieta rica em frutose, podendo ser pensado como uma nova ferramenta terapêutica no tratamento da superativação do eixo ECA/AT1r. / High-activation of ACE/AT1r axis is closely linked to metabolix syndrome and low-grade inflammation state. Fructose administration in water or in diet has been proposed as a model to study the high-activity of this axis and AT1r-related inflammatory pathways. In this view, we aimed to evaluate if GW501516 administration could decrease the high-activation of ACE/AT1r axis and consequently fructose damage. To this males mice C57Bl/6 were fed a high-fructose diet (47%) during eight weeks or standard-chow diet. After eight weeks, the groups were randomly divided to start treatment with GW501516, totaling four experimental groups. Animals treated with GW501516 presented an improvement of systolic blood pressure and in urinary parameters, as proteinuria and uric acid. Also was verified an improvement in plasmatic triglycerides and uric acid. In white adipose tissue, GW501516 was able to decrease the components of this axis and improved inflammation as well. In liver, it was not found differences in axis, but JAK2 phosphorylation AT1r-dependent was decreased and consequently it was found a diminished activations of hepatic stellate cells. In addition, proteins and genes related to &#946;-oxidation were increased with GW501516 and those related to lipogenesis de novo, were diminished, improving hepatic parenchyma. Kidneys presented an improvement of inflammation state, although it was not found differences in axis with treatment. Also, it was not found differences in gene and protein expression in relation to anti-oxidants proteins. These results show that short-administration of GW501516 could alleviate inflammatory effects and hepatic steatosis caused by high fructose diet, suggesting that GW501516 could be a new therapeutic option to treat the high activation of ACE/AT1r axis.

Os piretróides sintéticos, em especial os do tipo II, como a cialotrina, são extensivamente utilizados para o controle de uma ampla variedade de ectoparasitas que acometem os animais de produção. Entretanto, no Brasil e em outros países, sua utilização vai além da saúde animal, sendo utilizados também em saúde pública, no controle de diversos vetores, como é o caso do vetor da dengue, dentre outros. Visto que a cialotrina modifica a atividade de macrófagos peritoneais, o objetivo deste trabalho foi investigar os prováveis mecanismos através dos quais este piretróide modifica a atividade destas células. Os presentes resultados, analisados em seu conjunto, mostram de maneira inequívoca que a cialotrina tem um efeito direto e/ou indireto sobre a atividade de macrófagos peritoneais. Especificamente, observou-se neste trabalho que o praguicida causou em ratos: 1 ? marcação fos positiva em neurônios do núcleo paraventricular do hipotálamo (NPH), após a dose de 3,0 mg/kg/dia; 2 - diminuição do percentual e intensidade de fagocitose de macrófagos peritoneais ativados e avaliados por citometria de fluxo; 3 - diminuição dose-dependente da produção de nitrito (NO2); 4 ? diminuição do percentual e intensidade de fagocitose de macrófagos peritoneais ativados, em ratos adrenalectomizados e/ou tratados com metirapona (inibidor da síntese de corticosterona) e RU 486 (antagonista de receptores glicocorticóides) com a finalidade de modular os níveis de glicocorticóides, e tratados com 3,0 mg/kg/dia de cialotrina; 5 ? aumento dos níveis de noradrenalina hipotalâmica em animais tratados com a dose de 3,0mg/kg/dia de cialotrina; 6 - diminuição do percentual e intensidade de fagocitose, bem como diminuição da produção de nitrito de macrófagos peritoneais ativados, em ratos simpatectomizados químicamente com 6-OHDA; 7 - diminuição dose dependente do percentual e intensidade de fagocitose, bem como da produção de nitrito de macrófagos peritoneais ativados e tratados in vitro com 10 e 100 nM de cialotrina. No entanto, não observamos: 1 ? alterações na produção de nitrito realizada por macrófagos peritoneais ativados, em ratos adrenalectomizados e/ou tratados com metirapona e RU 486; 2 - alterações na viabilidade celular induzida pelo tratamento in vitro com a cialotrina na concentração de 10 e 100 nM e 3 ? alterações nos efeitos da cialotrina sobre a atividade de macrófagos tratados in vitro com os ligantes de receptores benzodiazepínicos periféricos. Em conjunto, os presentes dados mostram que a cialotrina interfere com a atividade de macrófagos por atuar indiretamente, através da ativação do eixo Hipotálamo-Hipófise-Adrenal (HHA), e/ou diretamente sobre os mesmos modulando sua atividade. É muito provável que o efeito resultante do tratamento in vivo com este praguicida esteja ligado à somatória destas ações / Synthetic pyrethroids, particularly those of type II, such as cyhalothrin, are extensively used in agriculture for the control of a broad range of ectoparasites in farm animals. However, in Brazil and some other countries, these pyrethroids have also been used in public health, for the control of insects that are known to be vectors of diseases such as dengue. Since it has been suggested that cyhalothrin alters activity of peritoneal macrophages, the objective of our study was to investigate the putative mechanisms for the changes induced by pyrethroid in these cells. The results presented here show, in an unequivocal manner, that cyhalothrin has a direct or indirect (or both) effect on the activity of peritoneal macrophages. We specifically observed in this work that this pesticide induced in rats: 1- Fos-positive immunostaining in neurons of the paraventricular nucleus of the hypothalamus (NPH), after 3.0 mg/kg/day; 2 ? a reduction in the percentage and intensity of phagocytosis by activated peritoneal macrophages, evaluated by flow cytometry; 3 ? a dose-dependent reduction in nitrite production (NO2); 4 ? a reduction in the percentage and intensity of phagocytosis by activated peritoneal macrophage from adrenalectomized rats treated or not with metirapone (inhibitor of corticosterone synthesis) or RU 486 (antagonist of glicocorticoids receptors) with the propose of modulating the levels of glicocorticoids, and treated with 3.0 mg/kg/day of cyhalothrin; 5 ? an increase in the hypothalamic levels of noradrenaline in rats treated with 3.0 mg/kg/day of cyhalothrin; 6 ? a reduction in the percentage and intensity of phagocytosis and also a decrease in the production of nitrite by activated peritoneal macrophages, after chemical sympatectomy with 6-OHDA; 7 ? a dose-dependent reduction of the percentage and intensity of phagocytosis, and also a decrement in nitrite production by activated peritoneal macrophages treated in vitro with 10 and 100 nM of cyhalothrin. However, we found no differences on: 1 ? nitrite production by activated peritoneal macrophages after adrenalectomy, treated or not with metirapone or RU 486; 2 ?cell viability of peritoneal macrophages treated in vitro with 10 and 100 nM of cyhalothrin, and 3 ? the effects of cyhalothrin on macrophage activity after in vitro treatment with peripheral benzodiazepine receptor ligands. Altogether, the present results show that cyhalothrin interferes with the activity of peritoneal macrophages by acting indirectly, via activation of the hypothalamus-pituitary-adrenal axis, or directly on these cells, altering their activity. As a matter of fact, it is quite possible that the results of in vivo cyhalothrin treatment on macrophage activity would be related to the combined effect of these direct and indirect influences