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.

<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.

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.

Orientador: Wílson Mello Júnior / Resumo: O carcinoma de bexiga urinária é o tipo de câncer mais prevalente no sistema urinário em seres humanos e apresenta altos índices de morbidade e mortalidade. Seu desenvolvimento está relacionado a diversos fatores de risco, como o hábito de fumar, a exposição a agentes químicos industriais e metais pesados e o estresse crônico. As experiências adversas no início da vida estão relacionadas à modulação de eixos hormonais importantes, como o hipotalâmicohipofisário-adrenal (HHA) e hipotalâmico-hipofisário-gonadal (HHG). Alterações na resposta do eixo HHA, componente central da resposta fisiológica ao estresse, levam a desordens fisiológicas, como aumento da proliferação celular, angiogênese e prejuízo da resposta imune, que podem ser responsáveis pelo aumento da susceptibilidade do indivíduo a uma série de doenças na vida adulta, incluindo o câncer. Paralelamente, alterações na fisiologia de hormônios esteroides como andrógenos, estrógenos e glicocorticoides, bem como seus receptores, também têm sido relacionadas ao desenvolvimento de enfermidades como o câncer de bexiga urinária. Em ratos, o comportamento materno funciona como um fator importante na modulação desses eixos hormonais, modificando suas respostas na vida adulta, e estudos recentes têm relacionado os cuidados recebidos por filhotes à incidência de tumores. O objetivo deste estudo foi avaliar a influência do comportamento materno e sua consequente modulação nos eixos HHA e HHG, no desenvolvimento do câncer de bexiga u... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The urinary bladder carcinoma is the most prevalent type of cancer in the urinary tract in humans, with high morbidity and mortality. Its development is related to several risk factors such as smoking, exposure to industrial chemicals and heavy metals and chronic stress. Adverse experiences early in life are related to modulation of important hormonal axes, as the hypothalamic-pituitary-adrenal (HPA) and hypothalamic-pituitary-gonadal (HPG). Changes in the response of the HPA axis, a central component of the physiological response to stress, lead to physiological disorders, such as increased cell proliferation, angiogenesis, and impaired immune response, which may be responsible for the increased susceptibility of the individual to a number of diseases in adult life, including cancer. Similarly, changes in the physiology of steroid hormones such as androgens, oestrogens and glucocorticoids and their receptors have also been related to the development of diseases such as bladder cancer. In rats, maternal behavior serves as an important factor in the modulation of hormonal axes, modifying their responses in adulthood, and recent studies have linked the care received by pups and the incidence of tumors. The aim of this study was to evaluate the influence of maternal behavior and its consequent modulation in HPA and HPG axes in the development of bladder cancer. The results showed that rats reared by the negligent mothers had a greater susceptibility to the development of chemica... (Complete abstract click electronic access below) / Mestre

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.

Em função da categoria dos estressores, vias neurais específicas são envolvidas e respostas distintas podem ser induzidas. A literatura tem reportado que o estrógeno (E 2 ) através de seus receptores de tipos (ER) e (ER) influencia a atividade do eixo hipotálamo hipófise adrenal (HPA). Além disso, há evidências de que o E2 exerça efeitos protetores em situação de choque hemorrágico. O objetivo deste trabalho foi verificar a participação dos receptores ER e ER na atividade do eixo HPA durante estresse hemorrágico. Foram utilizadas ratas Wistar ovariectomizadas que receberam injeções s.c. de DMSO (veículo), PPT (agonista ER) ou DPN (agonista ER), durante 3 dias. No segundo dia, as ratas foram canuladas para coleta seriada de sangue na manhã seguinte. Os animais receberam (controle) ou não (hemorrágicos) reposição imediata com salina. Os hormônios corticosterona (CORT), ocitocina (OT) e vasopressina (AVP) foram dosados por radioimunoensaio. Ao final do experimento, os ratos foram perfundidos e os cérebros processados para imuno-histoquímica de FOS, tirosina hidroxilase (TH) e hormônio liberador de corticotrofina (CRH). Nos animais tratados com veículo, a hemorragia gradual moderada aumentou a secreção de CORT, OT e AVP, a expressão de neurônios TH ativados na região A1C1 e de FOS no mpPVN. O PPT reduziu a secreção de CORT, na situação controle atuando no LC e mpPVN; e também após hemorragia atuando no LC, NTS, A1C1 e mpPVN. O DPN reduziu a secreção de CORT apenas após estresse hemorrágico atuando no LC, A1C1 e mpPVN. O PPT bloqueou o aumento da secreção de OT e aumentou a secreção de AVP, após hemorragia. O DPN, por sua vez, reduziu a concentração plasmática de OT e aumentou a concentração plasmática de AVP, independentemente da hemorragia. Em conclusão: o estrógeno pode exercer uma ação inibitória sobre a secreção basal de CORT somente através da ação do ER sobre o LC e mpPVN; a secreção de CORT aumenta em resposta à hemorragia gradual moderada e o estrógeno pode exercer um controle inibitório nessa resposta através de ER atuando sobre LC, NTS, A1C1 e mpPVN, bem como através de ER atuando sobre LC, A1C1 e mpPVN. / Depending on the stressors category, specific neural pathways are involved and different responses can be selected. It has been reported in the literature that estrogen (E2 ) can affect hypothalamus pituitary adrenal (HPA) axis activity through its receptors type (ER) and (ER). Moreover, there is evidence that E 2 has protecting properties after hemorrhagic shock. The aim of this work was to assess the participation of ER and ER on HPA axis activity during hemorrhagic stress. It was used ovariectomized Wistar rats that received s.c. injections of: DMSO (vehicle), PPT (ER agonist) or DPN (ER agonist), during 3 days. In the second day the rats were catheterized for serial blood collect in the next morning. Animals received (control) or not (hemorrhagic) immediate reposition with same volume of isotonic saline. The hormones corticosterone (CORT), oxytocin (OT) and vasopressin (AVP) were measured by radioimmunoassay. At the end of the experiment, animals were perfused and their brains were processed for immuno-histochemistry for FOS, tyrosine hydroxylase (TH) and corticotropin releasing hormone (CRH). In vehicle treated animals, the gradual hemorrhage enhanced CORT, OT and AVP secretion, TH activated neurons expression in A1C1 and FOS expression in mpPVN. PPT decreased plasma CORT in control situation acting on LC and mpPVN, and also after hemorrhage acting on LC, NTS, A1C1 and mpPVN. DPN reduced plasma CORT only after hemorrhagic stress acting on LC, A1C1 and mpPVN. PPT blocked the increase of OT secretion and increased AVP secretion, after hemorrhage. The agonist DPN reduced OT and increased AVP levels, despite hemorrhage. In conclusion: E2 can exert an inhibitory effect on CORT basal secretion only through ER action on LC and mpPVN; CORT secretion increases after gradual moderate hemorrhage and E2 inhibit this secretion through ER action on LC, NTS, A1C1 and mpPVN, as well through ER action on LC, A1C1 and mpPVN.

Dysregulation of the hypothalamo-pituitary-adrenal (HPA) axis activity has been commonly observed among breast cancer patients and has been linked to adverse health consequences. However, whether these alterations persist long after the cancer diagnosis has not been well-documented. In the first study, the diurnal cortisol rhythms and the cortisol stress response of breast cancer survivors who had completed all local and/or systemic adjuvant therapy with the exception of hormonal therapy were compared to those of women without a history of cancer. The Trier Social Stress Test was used to elicit a moderate stress response and the subjective levels of stress of participants were recorded using visual analog scales. The results indicate similar diurnal patterns in both groups; however, significant differences in stress reactivity were noted, with breast cancer survivors displaying a relatively flat profile following the acute stress induction. Subjective levels of psychological stress were similar in both groups, indicating that the subjective appraisal did not account for the blunted cortisol stress response. In the second study, the impact of the stressful life events that happened during the previous year on the cortisol stress response was analyzed in the same groups of participants. The frequency of stressful life events as well as their subjective impact was documented using the Life Experience Survey. Results suggest no group differences between the total number of stressful life events and their perceived effect. However, the number of stressful life events and their perceived impact correlated negatively with the peak cortisol concentration in breast cancer survivors. The results suggest that the cumulative effect of stressful life events contribute significantly to the low levels of cortisol reported in breast cancer survivors following a stressful situation. Together, these studies emphasize that breast cancer survivors are at risk of presenting a subtle alteration of their HPA axis activity when their system is challenged and that an accumulation of stressors plays a role in this dysregulation. These results reinforce the need for interventions intended to reduce the levels of psychological stress experienced by breast cancer survivors.

A life event is as an occurrence that involves a subsequent change in the life pattern of an individual (Holmes & Rahe, 1967). The current study investigated whether exposure to life events over the past year influenced hypothalamic-pituitary-adrenocortical (HPA) axis function in healthy children and adolescents, and explored whether sex, age, behavioural inhibition, trait anxiety, anxiety sensitivity, perceived parental bonding, and parental history of anxiety moderated this relationship. The sample included 147 healthy children and adolescents. Participants were administered Coddington’s Life Events Scale (CLES) and salivary cortisol was collected for the determination of the cortisol awakening response (CAR), diurnal cortisol, and cortisol reactivity to a laboratory stressor. Separate linear regression models were conducted for each cortisol profile. Results revealed that life events significantly predicted total CAR output, diurnal cortisol response, and cortisol reactivity to a laboratory stressor. Further, behavioural inhibition, trait anxiety, not having a parental history of anxiety, and paternal caring positively moderated some of the relationships between life events and cortisol profiles. Considering the physiological and psychological effects of early exposure to stress, this study is significant in understanding the impact of life events to improve the health of children and adolescents.

Understanding the physiological response of marine mammals to anthropogenic stressors can inform marine ecosystem conservation strategies. Stress stimulates release of glucocorticoid (GC) hormones, which increase energy substrate availability while suppressing energy-intensive processes. Exposure to repeated stressors can potentially affect an animal’s ability to respond to and recover from subsequent challenges. To assess the endocrine response of a marine mammal to repeated stressors, we administered adrenocorticotropic hormone (ACTH) to free-ranging juvenile northern elephant seals (Mirounga angustirostris; n=7) once daily for four days. ACTH administration induced significant, but transient (<24 h) elevation in circulating cortisol levels (p < 0.0001). These increases did not vary in magnitude between the first ACTH challenge on day 1 and the last challenge on day 4. In contrast, aldosterone levels remained elevated above baseline for at least 24 hours after each ACTH injection (p < 0.001), and responses were greater on day 4 than day 1 (p < 0.01). Total triiodothyronine (tT3) levels were decreased on day 4 relative to day 1 (p < 0.01), while reverse triiodothyronine (rT3) concentrations increased relative to baseline on days 1 and 4 (p < 0.001) in response to ACTH, indicating a suppression of thyroid hormone secretion. There was no effect of ACTH on the sex steroid dehydroepiandrosterone (DHEA). These results suggest that elephant seals are able to mount adrenal responses to multiple ACTH challenges. However, repeated stress results in facilitation of aldosterone secretion and suppression of tT3, which may impact osmoregulation and metabolism. We propose that aldosterone and tT3 are informative additional indicators of repeated stress in marine mammals.