<p>It has been suggested that environmental factors substantially contribute to the increased prevalence of peanut allergy in industrialized countries. Specifically, the role of disrupted barrier integrity in the gastrointestinal tract has been implicated in the development of food allergy. The use of non-steroidal anti-inflammatory drugs (NSAIDs), which increase intestinal permeability, for the treatment of pain and fever is prevalent in industrialized countries. Therefore, the first aim of this study was to determine whether treatment with indomethacin, a prototypical NSAID, would act in an adjuvant like manner to facilitate sensitization to co-administered peanut protein. Furthermore, we investigated whether indomethacin increases susceptibility to anaphylaxis following oral challenge with peanut in sensitized mice.</p> <p>First, a short model of cholera toxin-mediated sensitization to peanut was developed. Mice were given 1 mg of peanut protein and 5 μg of cholera toxin by oral gavage for 10 consecutive days. This resulted in a robust anaphylactic response and increased peanut-specific IgG1, but not IgE, two weeks following treatment. Mice exposed to peanut during a 10-day treatment with indomethacin (5 mg/kg on alternating days or 3.5 mg/kg daily) did not develop peanut-specific immunoglobulins or anaphylaxis following systemic challenge with peanut protein. Furthermore, treatment with two 5 mg/kg doses of indomethacin 24 and 1 hour before oral challenge did not facilitate anaphylaxis in peanut-sensitized mice. Therefore, we concluded that NSAID treatment is unlikely to play a role in the increased prevalence of peanut allergy, and that NSAID treatment does not increase susceptibility to peanut-induced anaphylaxis in sensitized mice.</p> <p>The second part of this study aimed to develop a short model of epicutaneous sensitization, and address the impact of epicutaneous exposure to peanut during infancy. We investigated the role of site of exposure, duration of exposure, epidermal integrity, strain and age in epicutaneous sensitization. 10 consecutive days of epicutaneous exposure to 20 μg of peanut protein through tape stripped skin induced robust anaphylaxis following i.p. challenge. Neither 7 days of exposure through tape stripped skin, nor 10 days of exposure through intact skin facilitated sensitization. The strength of sensitization was strain-dependent; peanut-specific IgG1 was increased equally in both C57BL/6, and BALB/c mice, and to a greater extent in C3H/HeJ mice. Peanut-specific IgE was increased in both BALB/c and C3H/HeJ mice three weeks following peanut exposure. Epicutaneous exposure beginning one day after birth did not facilitate the development of either peanut-specific IgE or IgG1 in BALB/c mice, or anaphylaxis following systemic challenge. Beginning exposure at 2 weeks of age resulted in peanut-specific IgE and IgG1 production, as well as a robust anaphylaxis following i.p. challenge. Therefore, the development of peanut allergy through epicutaneous exposure is age dependent, and exposure during the neonatal period results in a hyporesponsive response.</p> / Master of Science (MSc)

<p>BACKGROUND: The ontogeny of peanut allergy (PA) is poorly understood, and the treatment of its most severe manifestation, peanut-induced anaphylaxis (PIA), remains limited to rescue epinephrine. We argued that an untargeted metabolomic analysis would be a useful hypothesis-generating tool to identify novel biomarkers, mediators and possibly therapeutic targets in PA and PIA.</p> <p>METHODS: Models of PA and PIA used in this thesis involved either the oral administration of peanut along with cholera toxin or the topical application of peanut on tape-stripped skin. Liquid-chromatography mass-spectrometry (LC-MS) was performed to identify chemical changes in the serum of mice undergoing sensitization and anaphylaxis. Flow cytometry as well as <em>in vivo</em> gain-of-function and loss-of-function immunological studies were used to determine the biological significance of particular molecules in sensitization.</p> <p>RESULTS: LC-MS followed by multivariant analysis showed that the purine metabolism pathway was altered with elevated levels of uric acid (UA) in sensitized mice. UA depletion using allopurinol and uricase fully prevented the development of the allergic and anaphylactic phenotype. Conversely, administration of UA crystals, instead of cholera toxin or tape stripping along with peanut induced a typical allergic and anaphylactic phenotype. The effects of UA and UA crystals are likely a consequence of effects on the activation of resident dendritic cells. Post-challenge metabolic analysis also revealed a distinct metabolic signature in sensitized mice, highlighted by an increase in several metabolites such as histamine. Likewise, peanut allergic patients display a distinct metabolic profile after oral peanut challenge.</p> <p>CONCLUSION: We identified UA, released after damage to the mucosa and/or skin, as a critical alarmin that facilitates the development of Th2 immunity, specifically PA and PIA. Metabolomics analyses of either mice undergoing anaphylaxis or peanut allergic children subjected to a peanut oral challenge provided an extensive overview of metabolomic changes underlying these conditions. Further studies may lead to the identification of novel biomarkers and mediators.</p> / Master of Science in Medical Sciences (MSMS)

INTRODUCTION: Altered walking patterns in individuals with knee osteoarthritis (OA) are a key driver of disease; however, mechanisms underlying this relationship are not clear. Central sensitization is now recognized as an important contributor to pain experience in people with knee OA. In the presence of chronic pain and central sensitization, there are alterations in activity and connectivity across multiple brain regions (e.g., prefrontal cortex, primary motor cortex) that could influence dynamic control of gait. However, the association of central sensitization with gait patterns in knee OA has not been studied. Our objective was to investigate whether central sensitization in individuals with knee OA is associated with dynamic control of gait as assessed using wearable sensor-derived measures of gait complexity and dynamic stability. We hypothesized that a greater degree of sensitization would be associated with lower gait complexity and stability. METHODS: We used data from the 12th-year visit of the Multicenter Osteoarthritis Study, a longitudinal cohort study of individuals with and without knee OA. This was the first visit at which the relevant gait parameters were collected using wearable sensors. Individuals from this visit with valid data available for all measures contained in this analysis were included. Mechanical temporal summation (TS), an augmented response to repetitive mechanical stimulation, is a reliable and valid measure of central sensitization. To assess TS, participants’ pain ratings were assessed after a single wrist stimulus was applied with successively weighted probes until a pain rating of at least 4/10 was achieved (the highest weighted probe was used if that pain rating was not obtained), then was assessed again after 10 repeated stimuli at a rate of 1 Hz was applied with the same probe. A post-stimulation pain rating larger than the initial pain rating reflected facilitated TS. For gait analyses, participants completed 2 trials of a 20m walking test at their usual walking pace while wearing inertial sensors on their lower back and both feet. The lower back sensor was used to calculate sample entropy and Lyapunov exponent, both of which may provide a window into how dynamic control of gait is altered in disease. Sample entropy measures the inter-stride predictability or regularity of gait kinematics. Lower sample entropy represents a more regular or predictable (i.e., less complex) gait pattern and may reflect a reduced ability of the neuromotor system to adapt to ongoing changes experienced in daily walking. LE measures the inter-stride stability of gait kinematics. Large LE values represent more gait instability and may reflect a reduced ability of the motor system to recover from small perturbations. We averaged the gait outcomes across the two trials. Exposure variables were standardized. We determined the association between TS and gait parameters using linear regression while adjusting for age, sex, race, body mass index (BMI), depressive symptoms, education, and presence of radiographic knee OA. Analyses were not adjusted for pain or gait speed because we hypothesized that these impairments were on the causal pathway between our exposure and outcomes. RESULTS: Data from 2,179 participants (age = 62.4 ± 10.0 years, BMI = 29.1 ± 5.5 kg/m2, 56.3% female) were included in the analysis. One standard deviation (SD) increase in TS was associated with a -0.024 [95% Confidence intervals -0.038, -0.010] change in sample entropy (i.e., more regular gait) but not with a change in LE (-0.002 [95% Confidence intervals -0.008, 0.004]). CONCLUSION: Individuals with knee OA and central sensitization display a less complex or more predictable gait pattern, evidenced by lower sample entropy, likely reflecting a deterioration in dynamic control of gait. However, central sensitization was not associated with an individual’s ability to respond to natural fluctuations that occur in walking, as evidenced by a lack of association with LE. We propose a plausible mechanism in which central sensitization may cause a reduction in gait complexity via continuous nociceptor release of glutamate, NMDAR activation, and changes in motoneuron excitability and inhibition at multiple sites along the motor pathway. Although causality cannot be determined from this cross-sectional study, these findings provide support for an association between central sensitization and specific aspects of the walking pattern in people with knee OA. Interventions to mitigate central sensitization may be useful to improve gait quality in people with knee OA.

BACKGROUND:This paper proposes a novel model for homeopathic remedy action on living systems. Research indicates that homeopathic remedies (a) contain measurable source and silica nanoparticles heterogeneously dispersed in colloidal solution / (b) act by modulating biological function of the allostatic stress response network (c) evoke biphasic actions on living systems via organism-dependent adaptive and endogenously amplified effects / (d) improve systemic resilience.DISCUSSION:The proposed active components of homeopathic remedies are nanoparticles of source substance in water-based colloidal solution, not bulk-form drugs. Nanoparticles have unique biological and physico-chemical properties, including increased catalytic reactivity, protein and DNA adsorption, bioavailability, dose-sparing, electromagnetic, and quantum effects different from bulk-form materials. Trituration and/or liquid succussions during classical remedy preparation create "top-down" nanostructures. Plants can biosynthesize remedy-templated silica nanostructures. Nanoparticles stimulate hormesis, a beneficial low-dose adaptive response. Homeopathic remedies prescribed in low doses spaced intermittently over time act as biological signals that stimulate the organism's allostatic biological stress response network, evoking nonlinear modulatory, self-organizing change. Potential mechanisms include time-dependent sensitization (TDS), a type of adaptive plasticity/metaplasticity involving progressive amplification of host responses, which reverse direction and oscillate at physiological limits. To mobilize hormesis and TDS, the remedy must be appraised as a salient, but low level, novel threat, stressor, or homeostatic disruption for the whole organism. Silica nanoparticles adsorb remedy source and amplify effects. Properly-timed remedy dosing elicits disease-primed compensatory reversal in direction of maladaptive dynamics of the allostatic network, thus promoting resilience and recovery from disease.SUMMARY:Homeopathic remedies are proposed as source nanoparticles that mobilize hormesis and time-dependent sensitization via non-pharmacological effects on specific biological adaptive and amplification mechanisms. The nanoparticle nature of remedies would distinguish them from conventional bulk drugs in structure, morphology, and functional properties. Outcomes would depend upon the ability of the organism to respond to the remedy as a novel stressor or heterotypic biological threat, initiating reversals of cumulative, cross-adapted biological maladaptations underlying disease in the allostatic stress response network. Systemic resilience would improve. This model provides a foundation for theory-driven research on the role of nanomaterials in living systems, mechanisms of homeopathic remedy actions and translational uses in nanomedicine.

Over the last 5 decades, the retrospective pretest has been used in behavioral science research to battle key threats to the internal validity of posttest-only control-group and pretest-posttest only designs. The purpose of this study was to compare outcome measures resulting from four research design implementations incorporating the retrospective pretest: (a) pre-post-then, (b) pre-post/then, (c) post-then, and (d) post/then. The study analyzed the interaction effect of pretest sensitization and post-intervention survey order on two subjective measures: (a) a control measure not related to the intervention and (b) an experimental measure consistent with the intervention. Validity of subjective measurement outcomes were assessed by correlating resulting to objective performance measurement outcomes. A Situational Leadership® II (SLII) training workshop served as the intervention. The Work Involvement Scale of the self version of the Survey of Management Practices Survey served as the subjective control measure. The Clarification of Goals and Objectives Scale of the self version of the Survey of Management Practices Survey served as the subjective experimental measure. The Effectiveness Scale of the self version of the Leader Behavior Analysis II® served as the objective performance measure. This study detected differences in measurement outcomes from SLII participant responses to an experimental and a control measure. In the case of the experimental measure, differences were found in the magnitude and direction of the validity coefficients. In the case of the control measure, differences were found in the magnitude of the treatment effect between groups. These differences indicate that, for this study, the pre-post-then design produced the most valid results for the experimental measure. For the control measure in this study, the pre-post/then design produced the most valid results. Across both measures, the post/then design produced the least valid results.

A literatura recente sobre drogadicção trata a sensibilização produzida pela pré-exposição a estimulantes como um possível fator na aquisição, manutenção e recaída na dependência gerada por essas drogas. O objetivo desse trabalho foi verificar se a pré-exposição a dietilpropiona (DEP) e cafeína (CAF) sensibilizava ratos ao efeito reforçador da DEP. Quatro experimentos foram realizados. No Experimento 1 um grupo de ratos foi pré-exposto i.p. a DEP, enquanto outro grupo de ratos recebia veículo. Posteriormente foi medido o efeito agudo i.p. de três doses de DEP (1,0, 2,5 e 5,0 mg/kg) sobre a atividade motora. No Experimento 2, o procedimento de pré-exposição foi similar ao descrito para o Experimento 1. Posteriormente, foi medida a preferência condicionada de lugar (CPP) produzida por três doses de DEP (1,0, 2,5 e 5,0 mg/kg). O Experimento 3 constou de duas partes. Na primeira parte, ratos adultos foram pré-expostos a CAF i.p., enquanto que outro grupo recebia veículo. Posteriormente foi medida a CPP induzida por 2,5 mg/kg de DEP. Na segunda parte desse experimento, ratos adolescentes foram pré-expostos oralmente a CAF por 56 dias consecutivos, enquanto que outro grupo de ratos recebia água. Posteriormente foi medida a CPP induzida por 1,0 mg/kg de DEP. No Experimento 4, ratos adolescentes foram submetidos a um procedimento de pré-exposição similar ao descrito para a segunda parte do Experimento 3. Posteriormente foi medido o desempenho sob um esquema de razão progressiva (PR), em que os animais eram reforçados oralmente com uma solução de DEP. Os resultados do Experimento 1 mostraram que as doses de 2,5 e 5,0 mg/kg de DEP aumentaram a atividade motora. Também foi observado que, na dose de 5,0 mg/kg de DEP, esse efeito era maior para os animais que tinham sido pré-expostos, indicando que a CAF sensibilizou o efeito da DEP sobre a atividade motora. Os resultados do Experimento 2 indicaram que as doses de 2,5 e 5,0 mg/kg de DEP produziram CPP, mostrando com isso valor reforçador; porém, não foi observada sensibilização desse efeito. Os resultados do experimento 3 revelaram que nem a pré-exposição oral a CAF desde a adolescência até a idade adulta, nem a pré-exposição i.p. na idade adulta, sensibilizou o valor reforçador da DEP quando medido pelo modelo de CPP. Os resultados do Experimento 4 mostraram que a pré-exposição oral a CAF desde a adolescência até a idade adulta sensibilizou o valor reforçador da DEP quando medido pelo modelo de PR. Os resultados são discutidos em relação aos comportamento medidos pelos modelos usados para medir o valor reforçador da DEP ao potencial de dependência da DEP e à importância do consumo precoce de cafeína na vulnerabilidade à dependência de drogas. / Recent literature considers sensitization resulting from pre-exposure to psychostimulants as a possible factor in drug addiction acquisition, maintenance and relapse. The aim of this study was to verify if pre-exposure to the psychostimulants diethylpropion (DEP) and caffeine (DEP) would sensitize rats to the reinforcing value of DEP. Four experiments were conducted. In Experiment 1 a group of rats was pre-exposed to DEP while another group received vehicle. After that, the acute effect of i.p. DEP (1.0, 2.5 or 5.0 mg/kg) on motor activity was measured. A similar pre-exposure procedure was conducted in Experiment 2. After that, Conditioned Place Preference (CPP) induced by i.p. DEP (1.0, 2.5 or 5.0 mg/kg) was measured. Experiment 3 was subdivided in two parts. In the first one adult rats were pre-exposed to i.p. CAF while a control group received vehicle. After that, CPP induced by 2.5 mg/kg i.p. DEP was measured. In the second part of this experiment, adolescent rats were pre-exposed to oral CAF for 56 consecutive days, while a control group received water. After that, CPP induced by 1.0 mg/kg i.p. DEP was measured. In Experiment 4, adolescent rats were submitted to a pre-exposure procedure similar to that described for the second part of Experiment 3. After that, their responses reinforced by a DEP solution on a progressive ratio schedule (PR) were measured. Experiment 1 results showed that 2.5 and 5.0 mg/kg DEP increased motor activity. At 5.0 mg/kg this effect was more pronounced in pre-exposed animals. In Experiment 2, the doses of 2.5 and 5.0 mg/kg DEP induced CPP, demonstrating their reinforcing value. However, no sensitization effect was observed. Experiment 3 results revealed that neither pre-exposure to oral CAF from adolescence to adulthood nor i.p. CAF pre-exposure in adulthood sensitized DEP reinforcing value as measured by CPP. Experiment 4 results showed that oral CAF pre-exposure from adolescence to adulthood induced sensitization of DEP reinforcing value as measured by PR breaking point. Results are discussed in terms of the animal models to assess reinforcing value, the abuse potential of DEP and the relevance of early caffeine consumption on the vulnerability to drug dependence.

Orientador: Daniela Aparecida de Godoi Gonçalves / Resumo: Em adultos, a disfunção temporomandibular (DTM) frequentemente está associada com outras condições dolorosas, como a fibromialgia (FM). Os tender points (TPs), que são parte dos critérios diagnósticos para FM, indicam um aumento da sensibilidade à dor generalizada. Objetivos: Investigar a associação entre DTM dolorosa e o número de TPs em adolescentes, bem como a relação entre o número de TPs e o limiar de dor à pressão (LDP) para dor local, regional e generalizada, como possíveis indicativos de sensibilização central (SC). Ainda, explorar a associação entre DTM dolorosa e FM, investigar o impacto da DTM dolorosa na qualidade de vida (QV) do adolescente e a influência de outras condições comórbidas (dor no corpo, qualidade do sono, doenças sistêmicas, sedentarismo, obesidade, sintomas depressivos) nessa associação. Métodos: A amostra foi composta por 690 adolescentes brasileiros com (n=112) e sem DTM dolorosa (n=578), de 12 a 14 anos. A DTM dolorosa foi classificada de acordo com os Critérios de Diagnóstico de Pesquisa para DTM (RDC/TMD) - Eixo I. Os critérios estabelecidos por Yunus foram aplicados para avaliar a FM juvenil e os TPs. O questionário Pediatric Quality of Life InventoryTM foi aplicado para avaliar a QV. Estatísticas descritivas foram utilizadas para caracterizar os participantes, estratificando pela presença de DTM dolorosa. Testes de Mann-Whitney e χ2 foram aplicados para testar a associação entre DTM dolorosa e variáveis demográficas. Modelos de regressão for... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Temporomandibular disorder (TMD) in adults is often associated with other painful conditions, such as fibromyalgia (FM). The tender points (TPs), which are part of the diagnostic criteria for FM, indicate increased sensitivity to generalized pain. Aims: To investigate the association between painful TMD and the number of TPs in adolescents, as well as the relationship between the number of TPs and the pressure pain threshold (PPT) for local, regional and generalized pain, as possible indicative of central sensitization (CS). Also, to explore the association between painful TMD and FM, to investigate the impact of painful TMD on adolescent's quality if life (QoL) and the influence of other comorbid conditions (body pain, sleep quality, systemic diseases, sedentary lifestyle, obesity, depressive symptoms). Methods: The sample consisted of 690 Brazilian adolescents with and without painful TMD, aged 12-14 years old. Painful TMD was classified according to the Research Diagnostic Criteria for TMD (RDC/TMD) - Axis I. The criteria that were established by Yunus were applied to assess juvenile fibromyalgia and TPs. The generic Pediatric Quality of Life InventoryTM was applied to evaluate the QoL. Descriptive statistics were used to characterize the participants, stratifying by the presence of painful TMD. MannWhitney and χ2 -tests were applied to test the association between painful TMD and demographic variables. Regression models were used to estimate the association between painful TMD and TPs and to determine which additional predictive variables were associated with TPs. Regression analyses were performed to test the associations between the PPT values and number of TPs. Fisher's test was used to estimate the association between painful TMD and FM. Linear regression analyzes were performed with painful TMD and one of the other possible predictors... (Complete abstractelectronic access below) / Doutor

Submitted by Erika Demachki (erikademachki@gmail.com) on 2014-09-05T20:11:20Z No. of bitstreams: 2 Alves, Vinicius de Medeiros - 2014.pdf: 3082084 bytes, checksum: da4838d5fe24841429f43de84204d98a (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-09-05T20:11:20Z (GMT). No. of bitstreams: 2 Alves, Vinicius de Medeiros - 2014.pdf: 3082084 bytes, checksum: da4838d5fe24841429f43de84204d98a (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-03-17 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Repetitive exposure to a chemical agent can induce an immune reaction in inherently susceptible individuals that leads to skin sensitization. Although many chemicals have been reported as skin sensitizers, there have been very few reports analyzing the relationships between their molecular structure and the sensitization potential including the connection to skin permeability, which is widely considered to be mechanistically implicated in sensitization. In this study, we have compiled, curated, and integrated the largest publicly available datasets related to chemically-induced skin sensitization and skin permeability. Unexpectedly, no correlation between sensitization and permeability has been found. Predictive QSAR models have been developed and validated for both skin sensitization and skin permeability using a standardized workflow fully compliant with the OECD guidelines. The classification accuracies of QSAR models discriminating sensitizers from non-sensitizers were 0.68-0.88 when evaluated on several external validation sets. When compared to the predictions generated by the OECD QSAR Toolbox skin sensitization module, our models had significantly higher prediction accuracy for the same sets of external compounds as evaluated by Positive Predicted Rate and Negative Predicted Rate as well as Correct Classification Rate. We have also developed QSAR models of skin permeability measured quantitatively. Cross-species correlation between human and rodent permeability data was found to be low (r²=0.44); thus, skin permeability models were developed using human data only and their external accuracy was q²ext = 0.87 (for 62% of external compounds found within the model applicability domain). Skin sensitization models have been employed to identify putative chemical hazards in the Scorecard database of possible skin or sense organ toxicants that should be regarded as primary candidates for the experimental validation. / A exposição repetida a agentes químicos pode induzir a sensibilização da pele em indivíduos inerentemente suscetíveis e desencadear uma resposta imunológica exacerbada. Apesar de muitos compostos químicos estarem implicados na sensibilização da pele, existem poucos estudos analisando as relações entre a estrutura molecular e o potencial sensibilizador desses compostos, incluindo a conexão com a permeabilidade pela pele, a qual é referida como sendo primordial para o processo de sensibilização. Neste estudo foram compilados, integrados e preparados os maiores conjuntos de dados disponíveis publicamente relacionados tanto com a sensibilização da pele quanto à permeabilidade. Inesperadamente, não se encontrou correlação entre essas duas propriedades. Modelos de QSAR robustos e preditivos foram gerados e validados para ambas as propriedades usando um fluxo de trabalho totalmente complacente com as recomendações da OECD. As taxas de acerto dos modelos discriminaram estruturas sensibilizadoras de não sensibilizadoras com uma taxa de 0,68-0,88 de sucesso, quando avaliadas em vários conjuntos de validação externa. Quando comparados com o módulo de sensibilização da pele implementado na ferramenta QSAR Toolbox da OECD, os modelos tiveram baixa cobertura do espaço químico, mas precisão preditiva mais elevada para os mesmos conjuntos de compostos externos avaliados pelo valor de preditividade positiva e valor de preditividade negativa assim como pela acurácia balanceada. O coeficiente de correlação cruzada entre os dados de permeabilidade da pele humana e de roedores apresentou-se baixo (r²=0,44); assim, apenas o conjunto de dados de pele humana foi considerado para geração de modelos de permeabilidade, que apresentaram precisão externa de q²ext = 0,87 (para 62% dos compostos dentro do domínio de aplicabilidade). Modelos de sensibilização da pele foram empregados para identificação de toxicantes putativos no banco de dados de possíveis agentes toxicantes da Scorecard que podem ser considerados como candidatos para validação experimental.

Introdução: A fibromialgia (FM) é uma síndrome de dor crônica musculoesquelética difusa, cuja etiologia não está totalmente conhecida. A síndrome cursa com dor, alterações do humor e sintomas de ruptura do ritmo circadiano. Sabe-se que seu processo fisiopatogênico envolve um desbalanço entre os sistemas de modulação excitatório e inibitório da dor. A capacidade do sistema modulatório inibitório está enfraquecida, com hiperativação de neurônios e da neuroglia, constituindo um quadro de sensibilização central. Portanto, estudos adicionais são necessários para compreender a relação entre possíveis marcadores séricos da hiperativação neuronal, tais como o Brain Derived Neurotrophic Factor (BDNF) e a proteína S100 beta (S100B). Além disso, estudos que busquem opções terapêuticas com efeito em vias neurobiológicas alternativas, tais como a melatonina, uma indolamina com efeitos ressincronizador, analgésico, anti-inflamatório e em sistemas moduladores da dor, como o gabaérgico, opioidérgico e glutamatérgico. Objetivos: 1) Primário: Avaliar se os níveis séricos de BDNF e S100B teriam associação com a FM e se ambos os mediadores sorológicos poderiam ser associados com o limiar de dor à pressão. 2) Secundário: Testar o tratamento com melatonina isolada ou em combinação com amitriptilina é melhor que amitriptilina isolada para modificar o sistema modulatório da dor. Assim, para provar tais hipóteses, neste estudo foram quantificados a modulação condicionada da dor e níveis de BDNF sérico em pacientes que receberam tratamento com melatonina isolada ou associada com amitriptilina. Foi também testado se melatonina melhoraria os sintomas clínicos como dor, limiar de dor à pressão e qualidade do sono relacionado à FM. Métodos: Foram selecionadas pacientes com diagnóstico de FM de acordo com o American College of Rheumatology (ACR) 2010. No primeiro estudo, de desenho transversal, foram incluídas 56 mulheres com FM, com idades entre 18 e 65 anos. Foram avaliados o limiar de dor à pressão e dosagem sérica de BDNF e S100B. No segundo estudo, foram incluídas 63 pacientes com os mesmos critérios de inclusão descritos no estudo transversal. As pacientes foram randomizadas e receberam, ao deitar, amitriptilina (25mg) (n=21), melatonina (10mg) (n=21) ou melatonina (10 mg) + amitriptilina (25mg) (n=21), durante seis semanas. O sistema modulatório descendente da dor foi acessado pela modulação condicionada da dor, através da mensuração da escala numérica de dor (NPS(0-10)) durante aferição do limiar de dor ao calor. Resultados: O resultado do estudo transversal mostrou que BDNF e S100B séricos foram correlacionados. BDNF e S100B foram inversamente correlacionados com limiar de dor à pressão. BDNF sérico foi associado com limiar de dor à pressão, idade e transtorno obsessivo compulsivo, enquanto que S100B sérica foi apenas associada com limiar de dor à pressão. O ensaio clínico randomizado demonstrou que a melatonina aumentou a potência do sistema modulatório da dor inibitório e que a modulação condicionada da dor foi negativamente correlacionada com BDNF sérico. Conclusões: Os estudos desta tese demonstram que S100B e BDNF, ambos mediadores chave no processo de sensibilização central, foram inversamente correlacionados com o limiar de dor à pressão. BDNF sérico foi, ainda, inversamente correlacionado com a redução da dor. Portanto, a avaliação sérica de BDNF e S100B merece estudos adicionais para determinar seu potencial papel sinalizador no espectro da sensibilização central nessa doença. / Introduction: Fibromyalgia (FM) is a syndrome of chronic diffuse musculoskeletal pain whose etiology is not fully known. This syndrome causes pain, mood swings and symptoms of rupture of the circadian rhythm. Its pathophysiological process involves an imbalance between excitatory and inhibitory pain modulatory systems. The ability of inhibitory systems is weakened, providing a framework of central sensitization, with dysfunction in the descending pain modulatory system, hyper-activation of neurons and neuroglia. Therefore, additional studies are needed to understand the possible relationship between serum markers of neuronal hyperactivity, such as Brain Derived Neurotrophic Factor (BDNF) and S100B. Particularly, studies seeking therapeutic options with effect in neurobiological alternative pathways such as melatonin, a indolamine with resynchronization, analgesic, and anti-inflammatory effects and actions on the modulatory pain systems such as GABAergic, opiodergic and glutamatergic. Objectives: 1) Primary: Evaluate whether the serum levels of BDNF and S100B have association with FM and if both serological mediators could be associated with pressure pain threshold. 2) Secondary: To test the hypothesis that treatment with melatonin alone or in combination with amitriptyline is better than amitriptyline alone to modify the endogenous pain modulatory system. Thus, to prove these hypothesis, it was quantified the conditioned pain modulation and serum BDNF levels in FM patients receiving treatment with melatonin alone or in combination with amitriptyline. Also, it was tested whether melatonin would improve clinical symptoms such as pain, pressure pain threshold and quality of sleep related to FM. Methods: Patients with FM according to the American College of Rheumatology (ACR) 2010 were selected. In the first study, a cross-sectional design, 56 women aging 18-65 years old, with FM were included. It was evaluated the pressure pain threshold, and serum levels of BDNF and S100B. In the second study, 63 patients were included with the same inclusion criteria described in the cross-sectional study. Patients were randomized and received at bedtime amitriptyline (25 mg) (n = 21), melatonin (10 mg) (n = 21) or melatonin (10 mg) + amitriptyline (25 mg) (n = 21) for six weeks. The descending pain modulatory system was accessed by the conditioned pain modulation, measuring the numerical pain scale [NPS (0-10)] during the heat pain threshold. Results: On the cross-sectional study serum BDNF and S100B were correlated. Serum BDNF and S100B were correlated with the pressure pain threshold. Serum BDNF was associated with pressure pain threshold, age and obsessive compulsive disorder, while serum S100B was associated with pressure pain threshold, only. The randomized clinical trial showed that melatonin increased the efficacy of inhibitory pain modulatory system and the conditioned pain modulation was negatively correlated with serum BDNF. Conclusions: The studies of this thesis show that both key mediators of the central sensitization process, BDNF and S100B, were inversely correlated with the pressure pain threshold. They also showed that melatonin increased the inhibitory pain modutalory system. Furthermore, it emphasizes that serum BDNF was inversely correlated with pain reduction. Therefore, assessment of serum BDNF and S100B deserve further studies to determine their potential as a proxy for the central sensitization spectrum in FM.

This investigation was an analysis of the interaction of adolescent exposure to methylphenidate (MPH; trade name: Ritalin) on nicotine sensitization and conditioned place preference (CPP) in a rodent model and underlying mechanisms of this effect. Animals were treated IP with 1 mg/kg MPH or saline using a ―school day‖ regimen of five days on, two days off, from postnatal day (P) 28-50. During the final two weeks of MPH treatment, animals were either behaviorally sensitized to nicotine (0.5 mg/kg free base) or saline for 10 days, or conditioned to nicotine or saline using the CPP behavioral paradigm. In addition, three days after behavioral sensitization was complete, animals were analyzed for stress behavior using the forced swim stress behavioral test. In addition, 24 hours after post-test conditioning animals were analyzed for the effect of a clinically relevant dose of pre-exposed MPH (1mg/kg) and nicotine treatment on the expression of BDNF in the nucleus accumbens and dorsal hippocampus. Behavioral results revealed that adolescent pre-exposure to MPH blunted nicotine behavioral sensitization in both male and female rats during the first week of testing. However, MPH enhanced nicotine CPP in both adolescent male and female rats. Interesting, animals administered MPH demonstrated a significantly decreased latency to immobility in the forced swim stress behavioral test. In addition, pre-exposure to a 1 mg/kg dose of MPH appears to have sensitized the BDNF response to nicotine in females as compared to all other groups.