Considerando que processos cruciais de diferenciação e maturação das células dendríticas (DCs) são regulados pelo fator de transcrição nuclear kappa B (NF-&#954;B), nos propusemos a estudar esta via em DCs derivadas de monócitos de pacientes com câncer de mama, partindo da hipótese de que alterações desta via contribuam, nesses indivíduos, para a geração de DCs com fenótipo e função alterados, levando ao escape tumoral. A análise da presença de NF-&#954;B no núcleo de monócitos, DCs imaturas (iDCs) e maduras indicou que as pacientes falham em modular tal fator de transcrição, de modo que a quantidade de NF-&#954;B no núcleo de iDCs de pacientes supera os níveis encontrados em controles, fenômeno possivelmente decorrente de alterações nas proteínas inibidoras do NF-&#954;B em pacientes. Observou-se menor frequência de células CD86+, CD83+ e HLA-DR+ (p < 0,05) ao final das culturas das pacientes, e aumento na concentração de IL-8 no sobrenadante das culturas. Coletivamente, estes dados corroboram a hipótese formulada. / Considering that crucial processes of differentiation and maturation of dendritic cells (DCs) are regulated by nuclear factor kappa B (NF-&#954;B), we proposed to study this pathway in monocyte-derived DCs from breast cancer patients, based on the hypothesis that alterations in such pathway may contribute in these individuals to the generation of DCs having phenotypic and functional changes, leading to tumor escape. The analysis of the presence of NF-&#954;B in the nucleus of monocytes, immature and mature DCs indicated that patients fail to modulate this transcription factor, so that the amount of NF-&#954;B in the nucleus of iDCs from patients exceeds levels found in controls, a phenomenon possibly due to alterations in inhibitory proteins of NF-&#954;B in patients. It was also observed diminished frequency of CD86+, CD83+ and HLA-DR+ cells (p <0.05) at the end of the patients cultures, as well as increased IL-8 concentration in the culture supernatants. Collectively, these data support the hypothesis previously formulated.

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Considerando que processos cruciais de diferenciação e maturação das células dendríticas (DCs) são regulados pelo fator de transcrição nuclear kappa B (NF-&#954;B), nos propusemos a estudar esta via em DCs derivadas de monócitos de pacientes com câncer de mama, partindo da hipótese de que alterações desta via contribuam, nesses indivíduos, para a geração de DCs com fenótipo e função alterados, levando ao escape tumoral. A análise da presença de NF-&#954;B no núcleo de monócitos, DCs imaturas (iDCs) e maduras indicou que as pacientes falham em modular tal fator de transcrição, de modo que a quantidade de NF-&#954;B no núcleo de iDCs de pacientes supera os níveis encontrados em controles, fenômeno possivelmente decorrente de alterações nas proteínas inibidoras do NF-&#954;B em pacientes. Observou-se menor frequência de células CD86+, CD83+ e HLA-DR+ (p < 0,05) ao final das culturas das pacientes, e aumento na concentração de IL-8 no sobrenadante das culturas. Coletivamente, estes dados corroboram a hipótese formulada. / Considering that crucial processes of differentiation and maturation of dendritic cells (DCs) are regulated by nuclear factor kappa B (NF-&#954;B), we proposed to study this pathway in monocyte-derived DCs from breast cancer patients, based on the hypothesis that alterations in such pathway may contribute in these individuals to the generation of DCs having phenotypic and functional changes, leading to tumor escape. The analysis of the presence of NF-&#954;B in the nucleus of monocytes, immature and mature DCs indicated that patients fail to modulate this transcription factor, so that the amount of NF-&#954;B in the nucleus of iDCs from patients exceeds levels found in controls, a phenomenon possibly due to alterations in inhibitory proteins of NF-&#954;B in patients. It was also observed diminished frequency of CD86+, CD83+ and HLA-DR+ cells (p <0.05) at the end of the patients cultures, as well as increased IL-8 concentration in the culture supernatants. Collectively, these data support the hypothesis previously formulated.

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Genetic and environmental factors interact to determine the individual vulnerability to develop ethanol dependence. The neurobiological mechanisms underlying these processes are not fully understood. Endogenous opioid peptides have been suggested to contribute. Brain opioids mediate ethanol reward and reinforcement via actions on the mesocorticolimbic dopamine system. This thesis focuses on environmental factors and investigates the impact of the early-life environment on adult voluntary ethanol consumption. The possible involvement of opioid peptides in environmental influences on adult ethanol consumption was examined using an experimental animal model. Maternal separation with short 15 min separations (MS15) was used to simulate a safe environment whereas prolonged 360 min separations (MS360) simulated an unsafe environment. Control rats were subjected to normal animal facility rearing (AFR). The separations were performed daily from postnatal day 1 to 21. Long-term ethanol consumption was registered using a two-bottle or a four-bottle free-choice paradigm in adult male and female ethanol-preferring AA (Alko, Alcohol), ethanol-avoiding ANA (Alko, Non-Alcohol) and non-preferring Wistar rats. In addition, analyses of immunoreactive Met-enkephalin-Arg6Phe7 (MEAP), dynorphin B (DYNB) and nociceptin/orphanin FQ (N/OFQ) peptide levels were performed after maternal separation as well as after voluntary ethanol drinking. In male rats, MS15 was related to lower ethanol consumption and these rats preferred lower concentrations, whereas MS360 was associated with an increased risk for higher consumption and/or preference for higher ethanol concentrations. Differences in basal opioid levels were observed in MS15 and MS360 rats. Furthermore, the ethanol-induced effects on opioid peptides in adults were dependent on the early environment. Female rats, on the other hand, were less affected or unaffected by maternal separation both in terms of ethanol consumption and neurobiological effects. Taken together, voluntary ethanol drinking, preference for low or high ethanol concentrations and opioid peptides in brain areas related to reward and reinforcement, motivation and stress were influenced by postnatal maternal separation in a sex dependent manner. The early environment thus had profound impact on the adult brain and the individual propensity for high ethanol drinking. A deranged endogenous opioid system contributed to these effects and may act as a mediator for long-term environmental influence on voluntary ethanol consumption.

Drug dependence is a serious and increasing problem in our society, especially among adolescents. The use of the large variety of substances available can result in a range of physiological and psychological adverse effects on individuals and negative consequences on the society overall. Several different types of drugs induce neurotoxicological damages, which in turn can generate impairment in for example the reward system and affect cognitive parameters.  The drug gamma-hydroxybutyrate (GHB) is usually considered a harmless compound among abusers, but has now shown to be highly addictive. Furthermore, GHB can cause memory impairments in both humans and animals. On the contrary, growth hormone (GH) and its main mediator insulin-like growth factor 1 (IGF-1) have recently been suggested to improve memory and learning in several studies. The hormones exhibit certain neuroprotective capabilities and have also previously been demonstrated to reverse opioid induced apoptosis in hippocampal cells. These effects and the fact that GHB is shown to increase GH secretion, which attracted considerable attention among body builders, led us to initiate studies on GHB and its impact on relevant systems in the central nervous system (CNS). Thus, the main purpose of the present investigation was to elucidate some of the underlying mechanisms that could account for the effects exerted by GH and GHB in the CNS. We found that a) GH affects the density and functionality of GABAB-receptors and opioid receptors in the male rat brain, b) GHB induces cognitive deficits and down-regulates GABAB-receptors, c) GHB treatment creates an imbalance between the endogenous opioids Met-enkaphalin-Arg6Phe7 (MEAP) and dynorphin B and increases the levels of MEAP in regions of the brain that are associated with drug dependence, and d) GHB affects the expression of IGF-1 receptors but not the plasma levels of IGF-1. In conclusion, the present work demonstrates that GH interacts with both opioid and GABAB-receptors in the male rat CNS and that GHB has an impact on brain regions associated with cognition and the development of dependence. These observations may be of relevance in many aspects related to addiction and might be translated into humans.

There is an unmet medical need for the efficient treatment of neuropathic pain, a condition that affects approximately 10% of the population worldwide. Current therapies need to be improved due to the associated side effects and lack of response in many patients. Moreover, neuropathic pain causes great suffering to patients and puts an economical burden on society. The work presented in this thesis addresses SP1-7, (Arg-Pro-Lys-Pro-Gln-Gln-Phe-OH), a major metabolite of the pronociceptive neuropeptide Substance P (SP). SP is released in the spinal cord following a noxious stimulus and binds to the NK1 receptor. In contrast to SP, the degradation fragment SP1-7 is antinociceptive through binding to specific binding sites distinct from the NK1 receptor. The aim of this thesis was to investigate the impact of SP1-7 on neuropathic pain. To understand how SP1-7 exerts its effect, a series of N-truncated forms of the heptapeptide were biologically evaluated. A set of small high-affinity ligands was evaluated in animal models of neuropathic pain. To confirm a clinical relevance the levels of SP1-7 in human neuropathic pain were assessed incerebrospinal fluid (CSF) collected from neuropathic pain patients. The results showed that SP1-7 could alleviate thermal as well as mechanical hypersensitivity in three different animal models of neuropathic pain. C-terminal amidation was connected with increased efficacy. N-terminal truncation of SP1-7 indicated a necessity of five amino acids in order to retain biological effect. One small high-affinity ligand showed a significant anti-allodynic effect. CSF levels of SP1-7 in neuropathic pain patients were lower compared to controls. Taken together, these findings demonstrate that the formation of SP1-7 may be attenuated in neuropathic pain. C-terminal amidation and a majority of its amino acids are necessary for stability and permeability. Clearly, SP1-7 and SP1-7 mimetics with high affinity to the SP1-7 binding site ameliorate neuropathic pain-like behaviors in animal models of neuropathic pain. Overall, the findings presented in this thesis contribute to new knowledge regarding the role of SP1-7 and related analogues and fragments in neuropathic pain. In a future perspective, this could be essential for the development of efficient strategies for managing patients with neuropathic pain.

The abuse of anabolic androgenic steroids (AAS) is becoming increasingly common and may result in a range of physiological as well as psychological effects such as altered behavior in terms of increased aggression, cognitive dysfunction and addictive behavior. AAS comprise testosterone and its derivatives, of which nandrolone is one of the more common. Previous studies have shown nandrolone-induced effects in male rats on peptide levels within the Substance P (SP) system and the dynorphinergic system; these effects may be linked to some of the reported behavior alterations. The studies presented in this thesis aimed to investigate the mechanisms underlying these peptide alterations and also to further investigate neuropeptidergic effects attributed to nandrolone administration. The results display significant effects on the enzymatic conversion of SP and Dynorphin A into their bioactive metabolites SP(1-7) and Leu-enkephalin-Arg6, respectively, as a result of nandrolone treatment. More profound investigations on the dynorphinergic system displayed effects on the kappa opioid receptor density in various brain regions. There was also a significant increase in the expression of the gene transcript of prodynorphin in the hippocampus, a brain region associated with cognitive processes. In addition, impaired spatial learning and memory in the Morris water maze task following nandrolone administration was encountered. The results provide further understanding regarding neuropeptidergic mechanisms underlying AAS-induced behavioral effects.