Epidemiological studies in humans have revealed a relationship between altered development in utero and an increased incidence of pathophysiology during postnatal life. One of the mechanisms underlying this relationship is thought to be exposure to excess glucocorticoids during critical phases of brain development. The aim of the current set of studies was to determine the effects of prenatal stress during discrete developmental windows on behaviour and endocrine function in male and female guinea pig offspring. Guinea pigs were used as the model for these studies as they are a long-gestation species that give birth to neuroanatomically mature young and fetal brain development is well characterized. Pregnant guinea pigs were exposed to a high frequency strobe light during the period of rapid fetal brain growth or the period of rapid brain myelination. Pregnant guinea pigs were allowed to deliver normally and guinea pig offspring were tested for ambulatory activity, anxiety and hypothalamic-pituitary-adrenal (HPA) axis function. Male offspring whose mothers were exposed to stress during the period of rapid brain growth exhibited increased anxiety behaviour, increased basal plasma cortisol levels and decreased plasma testosterone levels. We found that replacing testosterone in these animals reversed the behavioural and endocrine differences. Male offspring whose mothers were exposed to stress during the period of rapid myelination exhibited an increased plasma cortisol response to activation of the HPA axis. Female offspring whose mothers were exposed to stress during the period of rapid brain myelination exhibited decreased ambulatory activity and a blunted salivary cortisol response to the stress of the strobe light, but only during the estrous phase of the reproductive cycle. Therefore, the current set of studies has demonstrated the effects of prenatal stress on behaviour and HPA axis activity are dependent on; 1) the timing of the prenatal stress and 2) the hypothalamic-pituitary-gonadal axis in both male and female offspring. These studies have begun to uncover the mechanisms underlying programming and provide the basis for continuing research in humans.

Nuestros resultados previos han mostrado que una sola exposición a un estímulo estresante de elevada intensidad como la inmovilización (IMO) causa una reducción de la respuesta del eje hipotalámico-pituitario-adrenal (HPA) al mismo, pero no a un nuevo, estímulo estresante y este efecto se incrementa con el tiempo transcurrido (días o semanas) entre las dos exposiciones. La menor respuesta al mismo estímulo sugiere que una exposición previa a la IMO es capaz de inducir un cierto nivel de adaptación frente al mismo estímulo. Por lo tanto, la adaptación observada en respuesta al estrés crónico podría ser consecuencia del tiempo transcurrido entre la primera y la última exposición al estrés y no de la repetición del estímulo. Los objetivos del presente trabajo han sido: (a) comparar los efectos a largo plazo de una sola exposición al nado o a la IMO con los efectos de la exposición repetida; (b) estudiar el papel de los glucocorticoides o el miedo generados durante la primera exposición a la IMO en la inducción de los efectos a largo plazo de una sola exposición al estrés. Se utilizaron ratas macho adultas como animales de experimentación y se analizaron los niveles plasmáticos de ACTH, corticosterona y glucosa.No observamos adaptación del eje HPA, pero sí de la glucosa y otras variables, al nado crónico. Una sola exposición al nado no causó efectos a largo plazo en la respuesta del eje HPA al mismo estímulo, en contraposición con lo observado con la IMO. Por ello, decidimos comparar los efectos de una exposición única o repetida a la IMO en animales intactos y en animales adrenalectomizados suplementados con corticosterona en el agua de bebida (ADX+B). En los intactos, los efectos de la exposición repetida a la IMO sobre el eje HPA fueron muy similares a los de una sola exposición, pero en los ADX+B, una sola exposición a la IMO no modificó la respuesta de la ACTH al mismo estímulo, lo que sugiere que los glucocorticoides no serían necesarios para la adaptación a la IMO crónica, pero estarían implicados en la inducción o en la expresión de los efectos a largo plazo. Para profundizar en el papel de los glucocorticoides en la inducción de los efectos a largo plazo se manipuló su efecto biológico durante la primera exposición a la IMO inhibiendo su síntesis con metirapona, bloqueando sus receptores con el RU38486 o potenciando su liberación con la administración de ACTH. Los resultados sugieren que los glucocorticoides podrían estar implicados, aunque sólo parcialmente, en la inducción de los efectos a largo plazo sobre el eje HPA. La manipulación de los niveles de miedo/ansiedad con el fármaco ansiogénico FG7142 antes de la exposición al nado o el ansiolítico diazepam antes de la exposición a la IMO no modificó los efectos a largo plazo del estrés per se, lo que sugiere que el miedo/ansiedad no es un factor importante en la inducción de los efectos a largo plazo. Aunque una sola exposición previa a la IMO protegió parcialmente de la anorexia asociada a una nueva exposición al mismo estímulo, ninguna manipulación experimental fue capaz de modificar el efecto del estrés previo, indicando que los mecanismos neuroquímicos implicados en la inducción de los efectos a largo plazo de la IMO sobre la ingesta y el eje HPA son distintos. La futura caracterización de la respuesta conductual a la IMO podría ser de gran valor para determinar si la exposición a la IMO es un modelo experimental del síndrome de estrés postraumático en humanos. / We have previously observed that a single exposure of rats to severe stressors such as immobilization in wooden boards (IMO) results in a desensitization of the hypothalamic-pituitary-adrenal (HPA) response to the same, but not a novel, stressor and this process enhances with time (days/weeks) elapsed between the two exposures to the stressor. The lower HPA response to the same stressor suggests that some degree of adaptation developed after a single exposure to IMO and therefore is unclear whether adaptation to chronic repeated IMO may be explained by the time elapsed between the first and the final exposure rather than by repetition of the stressful situation. Thus the purposes of the present work were: (a) to compare the long-term effects of single exposure to forced swimming (FS) or IMO with repeated exposure; (b) to study the role of glucocorticoids or fear generated during the first exposure to IMO on the induction of long-term effects of an acute exposure to the same stressor. Two-month-old male Sprague-Dawley rats maintained under controlled conditions were used. Plasma levels of ACTH, corticosterone and glucose levels were measured. We found no significant adaptation of the HPA axis to chronic FS, but did of glucose and other variables. A single exposure to FS did not cause long-term effects on the responsiveness of the HPA axis to the same stressor in contrast to that observed after IMO. We then studied the effects of single versus repeated exposure to IMO on the HPA response to the same stressor in both sham-operated and adrenalectomized rats maintained with corticosterone in their drinking saline (ADX+B). In sham rats, daily exposure to IMO had similar effects on the HPA axis that a previous single session. In contrast, in ADX+B rats, a reduction of the ACTH response to the stressor was observed in repeated but not single IMO rats. The present result suggest that the glucocorticoids are not mandatory for the development or adaptation of the HPA axis to chronic IMO, but they may be involved in the induction or expression of long-term effects of IMO on the HPA responsiveness to the same stressor. In order to better study the role of glucocorticoids in the induction of long-term effects to IMO, three different experimental approaches were used during the first exposure to IMO: (i) inhibition of corticosterone synthesis with metyrapone; (ii) blockade of glucocorticoid receptors with RU38486; and (iii) potentiation of glucocorticoids release with the concomitant administration of exogenous ACTH. Taken together, the data suggest that glucocorticoids play only a partial role in the induction of long-term effects of IMO on the HPA. To study the contribution of fear/anxiety generated during the first exposure to stress we study the effect of the administration of the anxiogenic drug FG7142 in rats exposed to a moderate stressor (FS) or that of the anxiolytic diazepam in rats exposed to IMO. No effect of drugs was observed, suggesting that fear/anxiety is not one of the factors involved in the induction of the long-term effects of stress on the responsiveness of the HPA axis. Although a previous exposure to IMO was able to induce a partial protection on the effects on subsequent IMO-induced anorexia, neither the manipulation of glucocorticoid levels or function nor the administration of anxiety-related drugs were able to modify this protective effect. These results indicate that neurochemical mechanisms inducing long-term effects of food intake are different from those inducing changes in the HPA axis. The characterization of the behavioural response to IMO may be of value to determine whether or not exposure to IMO is an animal model for post-traumatic stress disorder.

El presente trabajo se centra en el estudio de las diferencias individuales en la respuesta endocrina al estrés. Mientras que la mayoría de estudios abordan este tema utilizando cepas de roedores seleccionados genéticamente, nosotros estamos interesados en el estudio de la susceptibilidad al estrés utilizando poblaciones de ratas adultas genéticamente diversas (outbred) de la cepa Sprague-Dawley, con el objetivo de prevenir posibles sesgos genéticos.Datos previos de nuestro laboratorio han demostrado que la velocidad de recuperación de los niveles basales de ACTH y corticosterona tras una exposición al estrés es un buen marcador de la intensidad del estrés (García et al., 2000). Hasta el momento se había asumido que la respuesta inicial de la ACTH a un estímulo estresante era un buen marcador de la intensidad de estrés. Sin embargo, como hemos comprobado en el presente trabajo, puede darse el caso de que los niveles plasmáticos de ACTH no discriminen entre estímulos estresantes severos (Márquez et al., 2002), confirmando la importancia del estudio de la dinámica de liberación de las hormonas del eje HPA después de finalizar la exposición a la situación estresante. Los datos de este trabajo apoyan que estudiado en las condiciones adecuadas, el eje HPA puede ser un muy buen marcador de la intensidad del estrés. Basándonos en este enfoque, hemos descrito la dinámica de adaptación a un paradigma de estrés crónico, estudiando detalladamente tanto la dinámica de liberación hormonal a lo largo de los días, como diferentes variables que pueden proporcionarnos información sobre el impacto final del estrés crónico. Hemos podido observar que aunque las diferencias individuales en la respuesta al estrés van haciéndose menos pronunciadas a lo largo del protocolo de estrés crónico, la respuesta a la primera exposición al estímulo estresante predice en parte el impacto final sobre el eje HPA a nivel central (Márquez et al., 2004). El hecho de que el perfil de la respuesta a una primera situación estresante pueda ser un índice de susceptibilidad a padecer patologías asociadas al estrés nos ha llevado a intentar caracterizar más detalladamente los factores que pueden estar determinando esta respuesta a una primera exposición a una situación estresante. Como paso previo a la caracterización de factores pronóstico de la respuesta de estrés hemos estudiado la consistencia y estabilidad de la respuesta al estrés de las hormonas del eje HPA. Hemos observado que la liberación de ACTH y corticosterona es bastante consistente frente a diferentes estímulos estresantes de intensidad baja-moderada (ambientes nuevos), independientemente de las características particulares de cada ambiente, lo que sugiriere que la liberación de las hormonas del eje HPA puede ser considerada una característica individual (rasgo) de cada animal. Sin embargo, cuando se han comparado situaciones estresantes que difieren ampliamente en cuanto a características cualitativas e intensidad, esta consistencia es mucho menor. Puesto que la mayor parte de los datos en la literatura en cuanto a susceptibilidad al estrés y diferencias individuales han utilizado situaciones estresantes de intensidad moderada, decidimos iniciar el estudio acerca de qué rasgos conductuales podrían estar relacionados con la respuesta al estrés utilizando ambientes nuevos. Gracias a la estrategia de caracterización conductual que nos ha permitido estudiar independientemente la influencia del rasgo de ansiedad del de actividad/exploración en ambientes nuevos, hemos descrito que ni la ansiedad ni la perseverancia en la exploración de ambientes nuevos parecen afectar el nivel de activación del eje en estas situaciones. Sin embargo, el nivel de actividad/exploración que muestran los individuos en el periodo inicial de exposición a un ambiente nuevo está positivamente relacionado con la reactividad del eje HPA ante situaciones de estrés. / The present work is focused in the study of individual differences of endocrine response to stress. Whereas most studies in this field use genetically selected rodents, we are interested in the study of stress susceptibility using outbred adult rats (Sprague-Dawley), in order to prevent possible genetic biases.Previous results from our laboratory have shown that the speed of recovery of ACTH and corticosterone basal levels after stress is a good marker of stress intensity (García et al., 2000). To date, it have been assumed that initial ACTH response to stress is a good intensity marker. However, as we have demonstrated in the present work, ACTH levels may not discriminate between highly stressful situations (Márquez et al., 2002). These data suggest that the follow up of the physiological response to severe stressors for a period of time after the termination of exposure to the situation allow us to better discriminate between the stressors in terms of intensity and could be a good marker of the possible pathological impact of such stressors. The present work corroborates that HPA axis response to stress is a very good intensity marker, if studied in the optimal conditions.We have also described the adaptation pattern to a chronic stress protocol, studying the hormonal response dynamics over the days, and studying different measures that may give us information about the final impact of chronic stress. Whereas individual differences in hormonal response to stress eventually disappeared after chronic stress, the hormonal response to the first encounter to the stressor may predict the final impact of chronic stress on the activity of HPA axis (Márquez et al., 2004). As the pattern of response to the first encounter to the stressor may be an index of susceptibility to stress-related pathologies, we have been interested in characterise variables that could be determining the response to this first encounter to stressor. As a first step to characterisation of prognostic factors of stress response, we have assessed the reliability of HPA response to stress. Our data suggest a good reliability of individual differences in the responsiveness of the HPA axis to stressors, when low to middle stressors are studied (novel environments), suggesting that HPA axis activation may be considered as a consistent individual trait of animals. Nevertheless, when highly differing stressors are compared, a lower reliability is observed. As most of the literature concerning stress susceptibility and individual differences have used low intensity stressors, we decide to use novel environments to start our last objective: to assess which personality traits could be related to a different stress responsiveness. Using a behavioural characterization strategy that allow us to study independently the influence of anxiety and novelty seeking traits, we have described that neither anxiety nor perseverance to explore novel environments seems to affect HPA response to stress. Nevertheless, initial exploration level of novel environments is positively related to HPA reactivity to stress.

<p>Several studies have shown that children who grow up under adverse care giving conditions are prone to develop a broad spectrum of different problems, ranging from mild depression to severe psychosomatic pathology later in life. A carefully treated child develops a different attachment strategy and biochemical response than a maltreated child. Early adverse events seem to program the stress response to become either over or under reactive which in turn have the potential to alter brain development. Major consequences include reduced plasticity and abnormal frontal lobe activity. This review further investigates the emotional and cognitive development in children exposed to early life abuse or neglect, trying to get a comprehensive picture of different symptoms that might contribute to later psychopathology.</p>

Several studies have shown that children who grow up under adverse care giving conditions are prone to develop a broad spectrum of different problems, ranging from mild depression to severe psychosomatic pathology later in life. A carefully treated child develops a different attachment strategy and biochemical response than a maltreated child. Early adverse events seem to program the stress response to become either over or under reactive which in turn have the potential to alter brain development. Major consequences include reduced plasticity and abnormal frontal lobe activity. This review further investigates the emotional and cognitive development in children exposed to early life abuse or neglect, trying to get a comprehensive picture of different symptoms that might contribute to later psychopathology.

Goal: To determine whether the Cortisol Awakening Response (CAR) associates with treatment response and course in hospital for inpatients with major depressive disorder (MDD). Methods: The CAR was measured at admission and discharge in patients completing a four-week inpatient program for MDD. Self-report questionnaires were used to assess changes in depression, anxiety, and perceived stress. Results: Over the four week hospital stay measures of CAR reactivity (Delta, AUCi) decreased, but there was no significant correlation between the change in CAR reactivity and change in clinical symptoms. Cross-sectional measurements of the CAR reactivity at both admission and discharge were strongly correlated with the drop in depression scores in hospital. Furthermore, poor treatment responders had a significantly lower CAR reactivity at both admission and discharge than did good responders. Conclusion: Individuals with higher CAR reactivity at admission and discharge had the greatest reduction in depression over the course of treatment.

Goal: To determine whether the Cortisol Awakening Response (CAR) associates with treatment response and course in hospital for inpatients with major depressive disorder (MDD). Methods: The CAR was measured at admission and discharge in patients completing a four-week inpatient program for MDD. Self-report questionnaires were used to assess changes in depression, anxiety, and perceived stress. Results: Over the four week hospital stay measures of CAR reactivity (Delta, AUCi) decreased, but there was no significant correlation between the change in CAR reactivity and change in clinical symptoms. Cross-sectional measurements of the CAR reactivity at both admission and discharge were strongly correlated with the drop in depression scores in hospital. Furthermore, poor treatment responders had a significantly lower CAR reactivity at both admission and discharge than did good responders. Conclusion: Individuals with higher CAR reactivity at admission and discharge had the greatest reduction in depression over the course of treatment.

Submitted by Pós Imunologia (ppgimicsufba@gmail.com) on 2017-03-14T15:46:34Z No. of bitstreams: 1 dissertação DANIELLE MIMOSO.pdf: 901513 bytes, checksum: 7f7c98092741a35d39bc3f951c13e94a (MD5) / Approved for entry into archive by Delba Rosa (delba@ufba.br) on 2017-04-24T12:30:34Z (GMT) No. of bitstreams: 1 dissertação DANIELLE MIMOSO.pdf: 901513 bytes, checksum: 7f7c98092741a35d39bc3f951c13e94a (MD5) / Made available in DSpace on 2017-04-24T12:30:34Z (GMT). No. of bitstreams: 1 dissertação DANIELLE MIMOSO.pdf: 901513 bytes, checksum: 7f7c98092741a35d39bc3f951c13e94a (MD5) / FAPESB / O eixo (HPA) hipotálamo-pituitária-adrenal é ativado em resposta ao estresse.Estudos mostraram que, em algumas infecções os níveis de cortisol, corticosterona e de ACTH estavam aumentados, indicando uma ativação do eixo HPA. Os glicocorticoides são hormônios que afetam o metabolismo e a resposta imunitária de várias formas, incluindo a diminuição da produção de citocinas pró-inflamatórias. A leishmaniose é uma doença causada por protozoários do gênero Leishmania, sendo um grave problema de saúde em várias partes do mundo, incluindo Brasil. Infecção por Leishmania pode levar a um amplo espectro de manifestações clínicas: a leishmaniose cutânea (LC), leishmaniose disseminada (DL), leishmaniose mucosa (LM), leishmaniose cutânea difusa (DCL) ea leishmaniose visceral (LV). No presente trabalho, investigamos se os pacientes com LV tem mudanças nos níveis de ACTH e cortisol e se essas alterações estão relacionadas a alguns marcadores clínicos e/ ou imunológicos da doença. MATERIAL E MÉTODOS Foram avaliados um grupo de pacientes com LV (n = 37) e CTL (n = 34), com idades variando de 2 a 11 anos e índice de massa corporal (IMC) similares.Os Hormônios (ACTH e cortisol) foram quantificados utilizando tecnologia Luminex e citocinas (IL1β, TNF-α, IL-6, IL-8, IL-10, IL-12, IFN-γ) foram determinadas por citometria de fluxo (CBA). RESULTADOS E DISCUSSÃO Os indivíduos com LV apresentaram níveis mais altos de cortisol (14,79 ng / mL ± 11,13) e ACTH (9.040 pg / mL ± 26,42) que os controles (12,00 ng / mL ± 1,831, P = 0,0004 e 1,880 pg / mL ± 48,08, P <0,0001, respectivamente) .O cortisol teve correlação positiva com o tempo de doença (r = 0,4105, P = 0,04), frequência respiratória (r = 0,44 P = 0,015) e IL-6 (r = 0,389, P = 0,0405). O ACTH teve correlação positiva com TNF-α (r = 0,3769, P = 0,040) e IL-1β (r = 0,3762, P = 0,0405). CONCLUSÕES: Nossos resultados indicam que os pacientes com LV tem uma ativação do eixo HPA e que esta pode ser associada com a progressão da doença. A elevação de algumas citocinas como a IL-6 pode estar envolvida nesta ativação e aumento do cortisol. / INTRODUCTION The Hypothalamic-Pituitary-Adrenal (HPA) axis is activated in response to stress. Studies have showed that in some infections there is elevation of cortisol or corticosterone and ACTH, indicating an activation of HPA axis. Glucocorticoids are hormones that affect the metabolism and immune response in several ways, including decreased production of proinflammatory cytokines. Leishmaniasis is a disease caused by protozoans of the genus Leishmania that is a serious health problem in several parts of the world including Brazil. Leishmania infection can lead to a wide spectrum of clinical manifestations: cutaneous leishmaniasis (CL), disseminated leishmaniasis (DL), mucosal leishmaniasis (ML), diffuse cutaneous leishmaniasis (DCL) and visceral leishmaniasis (VL). In the present work, we investigated whether patients with VL have changes in ACTH and cortisol levels and if these alterations are related to some clinical or immunological markers of disease. MATERIAL AND METHODS We evaluated a group of patients with VL (n=37) and healthy volunteers (n=34) with ages ranging from 2 to 11 years and similar body mass index (BMI). Hormones (ACTH and cortisol) were quantified using multiplex bead-based Luminex technology and cytokines (IL1β, TNF-α,IL-6,IL- 8,IL-10,IL-12,IFN-γ) were determinate by cytometric beads array (CBA). RESULTS AND DISCUSSION Individuals with VL exhibited higher levels of cortisol (14.79 ng/mL ±11.13) and ACTH (9.040 pg/mL±26.42) than controls (12.00 ng/mL±1.831, P=0.0004 and 1.880 pg/mL±48.08, P<0.0001, respectively).Levels of cortisol positively correlated with time of disease (r= 0.4105, P=0.04), respiratory frequency (r=0.44 P=0.015) and IL-6 (r=0.389, P=0.0405). ACTH positively correlated with TNF-α (r=0.3769, P=0.040) and IL- 1β (r=0.3762, P=0.0405). CONCLUSIONS: These results indicate that patients with VL have an activation of the HPA axis and that this activation could be associated with the outcome of disease. The elevation of some cytokines as IL-6 could be involved in the HPA activation and increase of cortisol.

Submitted by Everaldo Pereira (pereira.evera@gmail.com) on 2017-06-29T19:12:55Z No. of bitstreams: 1 dissertacao_atual_robson_2012_.pdf: 3646252 bytes, checksum: f996d44c7803422f675456f68434f40c (MD5) / Made available in DSpace on 2017-06-29T19:12:55Z (GMT). No. of bitstreams: 1 dissertacao_atual_robson_2012_.pdf: 3646252 bytes, checksum: f996d44c7803422f675456f68434f40c (MD5) / O pioneirismo da exploração de petróleo vivido pela Baía de Todos os Santos (BTS) deixou um grande passivo ambiental, sentido pelos ecossistemas locais sob a forma de contaminação por hidrocarbonetos de petróleo. Atualmente diversas técnicas lidam com essa problemática, dentre as quais, a biorremediação que é uma técnica aplicada para amenizar diversos impactos ambientais, dentre eles os gerados por derivados do petróleo, através da introdução de microorganismos no ambiente, ou criando condições ótimas para os mesmos se desenvolverem. O presente estudo objetivou avaliar a degradação de biomarcadores saturados e aromáticos através da técnica do bioestímulo em sedimento de manguezal impactado por atividades petrolíferas, testando a potencialidade de dois fertilizantes (NPK e OSMOCOTE) como agentes aceleradores da biodegradação dos componentes de petróleo, sob condições experimentais. A metodologia utilizada e testada no experimento baseouse no monitoramento do processo de biorremediação em aquários com água do rio São Paulo e sedimento contaminado por óleo, realizado durante 90 dias de simulação, ocorrido entre os meses de setembro e dezembro de 2008. Para o controle da simulação os parâmetros físico-químicos foram monitorados ao longo dos 90 dias da simulação, os teores de nitrogênio e fósforo foram monitorados mediante coletas de sedimento extraídos das provetas dos aquários de simulação e analisados no Laboratório de Estudos do Petróleo (LEPETRO). O óleo presente no sedimento foi extraído em soxhlet e posteriormente analisado por cromatografia liquida e gasosa. O resultado dos parâmetros físico-químicos mostrou condições favoráveis para o processo de biorremediação durante todas as fases de investigação e os nutrientes mantiveram dentro de limites adequados ao desenvolvimento dos microorganismos. Os biomarcadores saturados apresentaram indícios mais proeminentes de degradação ao longo da simulação, principalmente para os terpanos (m/z=191) e os esteranos (m/z=217). Os fragmentogramas (m/z= 170), (m/z=192), (m/z=228) e (m/z=231) analisados para o sedimento coletado no final da simulação apresentaram maiores abundância relativa dos picos, isso pode fornecer algumas conclusões, tais como, a possibilidade de degradação de compostos mais complexos que possam gerar frações de intermediária complexidade. Pelos resultados apresentados ao longo do experimento o nutriente NPK foi mais eficiente principalmente para as frações dos saturados. Sendo o nutriente OSMOCOTE recomendado para processos de biorremediação de compostos de alta complexidade (HPA de alto peso molecular e composto NSO), pois o tempo de resposta desse nutriente é mais lento, porém, é capaz de fornecer nutrientes por muitos meses. Como recomendação para futuros experimentos na região norte da BTS seria importante que fossem contemplados também as frações dos compostos NSO, pois a complexidade dessas substâncias tem que ser avaliada em qualquer processo de biorremediação. / ABSTRACT - The pioneer in oil exploration in Todos os Santos Bay (BTS) left a huge environmental liability, still sense by local ecosystems as (in the form of) contamination by petroleum hydrocarbons. Currently several techniques deal with this problem among which the bioremediation is a technology used to mitigate several environmental impacts, including those generated by petroleum, through the introduction of microorganisms in the environment or creating optimum conditions for them to develop. This study aimed to evaluate the degradation of saturated and aromatic biomarkers (PAH) by the technique of biostimulation in mangrove sediments impacted by oil activities, testing the potential of two fertilizers (NPK and OSMOCOTE) as agents to accelerate the biodegradation of oil, under experimental conditions. The methodology used and tested in the experiment was based on monitoring the process of bioremediation in aquariums with São Paulo river water and sediment contaminated by oil carried out during 90 days of simulation which occurred between September and December 2008. To control the simulation physicalchemical parameters were monitored over the 90 days of simulation and the nitrogen and phosphorus content were monitored by collection of sediment from the aquarium simulation and then analyzed in the Laboratory (LEPETRO). The oil present in the sediment was extracted with soxhlet and subsequently analyzed by liquid chromatography and gas. The result of the physicochemical parameters showed favorable conditions for the bioremediation process during all phases of research and nutrients remained within suitable limits for the development of microorganisms. The biomarkers saturated showed most prominent signs of degradation during the simulation, mainly for terpanes (m/z = 191) and steranes (m/z = 217). The fragmentograms (m/z = 170), (m/z = 192), (m/z = 228) and (m/z = 231) analyzed of the sediment collected at the end of the simulation showed higher peaks of relative abundance, that can provide some conclusions, such as the possibility of degradation of complex compounds can generate some fractions of intermediate complexity. For the results presented during the experiment the nutrient NPK was more efficient especially for the saturated fractions. The OSMOCOTE was the nutrient recommended for bioremediation processes of highly complex compounds (PAHs of high molecular weight and NSO compounds) because the response time of this nutrient is slower however is able to provide nutrients for many months. As a recommendation for future experiments in the northern region of the SBT would be important were covered the fractions of NSO compounds, because the complexity of these substances have to be evaluated in any process of bioremediation.

Os Transtornos por Uso de Substâncias (TUS) são propostos pela Associação Norte-Americana de Psiquiatria e englobam diversas dependências químicas, incluindo o tabagismo e a dependência de crack. O uso de crack tem aumentado consideravelmente nos últimos anos, segundo dados de levantamentos brasileiros e mundiais. Com mecanismo de ação semelhante ao da cocaína, porém com efeito mais rápido, o crack é uma droga que causa grave prejuízo a seus usuários e conduz à dependência em pouco tempo. O tabagismo também é uma condição bastante prevalente no mundo inteiro, sendo que os prejuízos relacionados a esse transtorno envolvem principalmente o desenvolvimento de doenças respiratórias e câncer. Diversos trabalhos têm buscado identificar fatores de suscetibilidade a esses transtornos, sendo que estudos de associação envolvendo genes do sistema de resposta a estresse vêm ganhando importância. O eixo Hipotálamo- Pituitária-Adrenal (HPA) é o sistema de resposta a estresse mais conhecido em humanos e alvo do presente trabalho, uma vez que o funcionamento dessa cascata de sinalização da produção de cortisol pode desempenhar um papel relevante no desenvolvimento de psicopatologias. Além disso, o eixo HPA pode ser considerado um mediador da relação entre situações adversas na infância e o surgimento de TUS. O objetivo desse trabalho, portanto, é investigar o papel de polimorfismos no gene CRHR1 (que codifica o receptor tipo 1 do hormônio liberador de corticotrofina – CRH, hormônio inicial do eixo HPA) sobre o desenvolvimento e características clínicas da dependência química. Para isso, foram realizados dois estudos, envolvendo amostras de mulheres. No primeiro artigo, a partir de três amostras – (1) 136 usuárias de crack, (2) 55 tabagistas e (3) 262 voluntárias saudáveis – foram identificadas diferenças de freqüências alélicas e genotípicas do SNP rs12944712; além disso, a análise de haplótipos envolvendo tal SNP e o rs110402 revelou maior frequência do haplótipo contendo o alelo G nos dois polimorfismos nos grupos de usuárias de crack e tabagistas. No segundo artigo, foi avaliado o efeito da interação geneambiente envolvendo os mesmos polimorfismos e situações adversas na infância – estimada pela escala Childhood Trauma Questionnaire (CTQ) –sobre características clínicas da amostra de usuárias de crack, como craving e gravidade da dependência – estimados, respectivamente, pelas escalas Cocaine Selective Severity Assessment (CSSA) e Addiction Severity Index (ASI6). A análise não revelou uma interação significativa. A presente dissertação, portanto, revela um efeito do gene CRHR1 sobre o desenvolvimento de TUS, embora esse efeito pareça não se manifestar sobre a gravidade. Mais estudos são necessários visando esclarecer o real efeito do gene e do eixo HPA como um todo sobre os fenótipos e comportamentos aditivos, bem como os mecanismos subjacentes a essa relação. / The Substance Use Disorders (SUD) are proposed by the American Psychiatric Association and comprise several addictions, including tobacco and crack cocaine smoking. The prevalence of crack cocaine dependence has grown considerably, according to survey data from Brazil and worldwide. With mechanism of action similar to cocaine but with faster effect, crack is a drug that causes severe impairments to its users and leads to dependence in a short time. Tobacco smoking is also a highly prevalent condition worldwide, and the impact related to this disorder includes cancer and respiratory diseases. Several studies have attempted the identification of susceptibility factors to these disorders, and association studies involving stress response system genes have been performed. The Hypothalamic-Pituitary-Adrenal (HPA) axis is the best known stress response system, and the signaling cascade production of cortisol may play a role in the development of psychopathology. In addition, HPA axis can be considered a mediator of the relationship between early life adversity and SUD. The aim of this study is to investigate the role of polymorphisms in CRHR1 gene (encoding the corticotrophin releasing factor – CRH – receptor type 1, an initial factor in HPA axis response) on the development and clinical traits of drug dependence. Two studies were conducted on women samples. In the first article, three groups were compared - (1) 136 crack smokers, (2) 55 tobacco smokers and (3) 262 healthy volunteers. Differences in allele and genotype frequencies of the SNP rs12944712 were identified; in addition, the haplotype analysis involving this SNP and rs110402 revealed a higher frequency of the haplotype containing G allele in both polymorphisms in the groups of nicotine dependents and crack smokers. In the second article, we evaluated the effect of a gene-environment interaction involving the same polymorphisms and childhood adversity - estimated by Childhood Trauma Questionnaire (CTQ) scale – on clinical characteristics of the sample of crack users, such as craving and severity of dependence - estimated respectively by the Cocaine Selective Severity Assessment (CSSA) and Addiction Severity Index (ASI6) scales. The analysis did not reveal a significant interaction. This dissertation thus shows an effect of CRHR1 genepolymorphism on the development of SUD, although this effect does not seem to impact disorder severity. More studies are needed in order to clarify the actual effect of the gene and of the HPA axis as a whole on addictive behaviors.