Made available in DSpace on 2017-07-10T14:17:20Z (GMT). No. of bitstreams: 1 Lana_Branai.pdf: 2297634 bytes, checksum: 8dbbaa1b059e5e38dcdd38ba4a303557 (MD5) Previous issue date: 2016-03-04 / Nowadays, the search for beauty, has caused health problems associated with the use of anabolic androgenic steroid (AAS) and even strenuous exercise, and the male reproductive system is the subject of studies to be sensitive to changes in the concentration of this type of hormone; these changes may also occur with increasing age, causing changes in androgen-dependent organs, like the epididymis. This study aimed to verify if treatment with AAS associated or not to exercise, in Sprague-Dawley rats alters the morphology of the epididymis in adult rats and its chronic effects in the elderly. The training was conducted by jumping into the water, weighing overloading, being considered as resistance exercise in water. The AAS administration occurred by intramuscular injection of nandrolone decanoate (10 mg / kg / week). Epididymal samples were subjected by histological routine of hematoxylin and eosin for morphological and morphometric analysis. It was analyze all parts of the epididymis (initial segment, caput, corpus and cauda) of 56 Sprague-Dawley rats, virgins, with 13 weeks old, divided into eight groups with seven animals each: GC - adults and sedentary; GCi - elderly and sedentary; GN - adults, sedentary treated with AAS; GNi - elderly, sedentary treated with AAS; GE - adults treated with exercise; GEi - elderly treated with exercise; GNE - adults, exercise and treatment with AAS; GNEi - elderly, exercise and treatment with AAS. The results show significant changes in duct diameters; the GE was lower when compared to other groups, and the groups that were used anabolic steroids, had a larger diameter than the other, and these changes occurred mainly in the initial segments. The epithelial height in the initial segment was also considered higher in the groups that administered AAS. The elderly groups tend to return to normal, increasing the parameters of epithelial height and diameter when these were decreased, and lowering them when they were enlarged when compared with the related adult group (with common variable), except at the tail of GNEi. As the results of this study, we can conclude that both treatment in adulthood, with exercise and the use of AAS, changes morphometric and morphological parameters of the epididymis, and its chronic effects can be diminished with age. / Atualmente, a procura pela beleza estética, tem causado problemas de saúde associados ao uso de esteroides androgênicos anabolizantes (EAAs) e até mesmo a exercícios físicos intensos. O sistema genital masculino é alvo de estudos por ser sensível a mudanças na concentração desse tipo de hormônio; essas alterações também podem ocorrer com o aumento da idade, influenciando órgãos androgênio-dependentes, como o epidídimo. Nesse sentido, objetivou-se com esse estudo, verificar se o tratamento com EAAs associado ou não ao exercício físico resistido, em ratos Sprague-Dawley, altera a morfologia do Epidídimo em ratos adultos e seus efeitos a longo prazo em idosos. Foram analisadas todas as porções dos epidídimos (segmento inicial, cabeça, corpo e cauda) de 56 ratos, virgens, da linhagem Sprague-Dawley (com 13 semanas de vida ao iniciarem o experimento), separados em oito grupos com sete animais cada: GC - adultos e sedentários; GCi - idosos e sedentários; GN - adultos, sedentários, tratados com EAA; GNi - idosos, sedentários, tratados com EAA; GE - adultos tratados com exercício; GEi - idosos tratados com exercício; GNE - adultos, exercício e tratamento com EAA; GNEi - idosos, exercício e tratamento com EAA. O treinamento realizado foi exercício resistido em meio aquático (através de saltos na água, com sobrecarga) com duração de oito semanas (três vezes na semana). A administração de EAAs ocorreu pela aplicação intramuscular de Decanoato de Nandrolona (10 mg/kg/semana) também durante oito semanas (duas vezes na semana). Amostras epididimárias passaram pela rotina histológica de hematoxilina e eosina para análise morfológica e morfométrica. Os resultados mostraram alterações significativas nos diâmetros de ductos, sendo que o GE foi menor quando comparado ao controle, e, nos grupos em que foram utilizados anabolizantes, tiveram diâmetro aumentado significativamente, e essas alterações ocorreram principalmente nos segmentos mais iniciais. A altura epitelial, no segmento inicial, também foi maior nos grupos em que foi utilizado anabolizante. Os grupos idosos tenderam a retornar a normalidade, aumentando os parâmetros de altura e diâmetro quando esses estavam diminuídos, e diminuindo-os quando estavam aumentados em comparação ao grupo adulto relacionado (com variável em comum), a não ser na cauda, do GNEi, em que houve aumento significativo. Achados morfológicos indicaram presença de debris celulares em lúmen (GN e GNEi) e infiltrados intersticiais (GN, GNE, GE e GCi). Conforme os resultados desse estudo, pôde-se concluir que tanto o tratamento na fase adulta com exercício, quanto à utilização de EAAs altera parâmetros morfométricos e morfológicos do epidídimo, e que seu efeito crônico pode ser diminuído com a idade.

<p>A new group of anabolic androgenic steroid (AAS) users has developed during the last two decades. This group consists primarily of young men interested in improving their physical appearance. Within this group, AAS are sometimes used together with other illicit drugs, alcohol and nicotine. Brutal and violent crimes have been committed under the influence of AAS, possibly because of AAS psychiatric side effects, ranging from increased aggression and psychosis to depression. Unfortunately, the biochemical mechanisms behind these effects are poorly understood.</p><p>In this thesis we used an animal model to study biochemical and behavioural effects of chronic AAS treatment (15 mg/kg/day of nandrolone decanoate for 14 days). The effect on the endogenous opioid peptides and the expression of immediate-early gene protein Fos in various brain regions were studied using radioimmunoassay and immunohistochemistry, respectively. In addition, we studied AAS effect on voluntary alcohol consumption and defensive behaviours, including aggression. The results show that AAS enhance endogenous opioid activity and Fos expression in brain regions regulating reward, aggression and disinhibitory behaviours. An imbalance between two opioid systems with generally opposing effects, the enkephalins with euphoric and the dynorphins with dysphoric effects, was also found. This implies that AAS alter the ability to maintain a stable state of mind and the response to other drugs of abuse. The AAS pre-treated animals enhanced their alcohol intake, were more aggressive and showed lower fleeing and freezing reaction than the controls. In addition, AAS enhanced amphetamine-induced aggression when the amphetamine was given three weeks after the last AAS injection.</p><p>The behavioural and biochemical results found in this thesis, support the hypothesis that use of AAS might lead to the development of dependence and may induce changes in the brain leading to disinhibitory behaviours.</p>

<p>The neurosteroids constitute a group of powerful hormones synthesized and acting in the central nervous system. They participate in a number of important central processes, such as memory and learning, mood and neuroprotection. Their effects emerge from rapid interactions with membrane bound receptors, such as the N-methyl-D-aspartate (NMDA) receptor, the gamma-amino-butyric acid receptor and the sigma 1 receptor. The mechanisms of action are separate from classical genomic interactions. </p><p>The aims of this thesis were to identify and characterize the molecular mechanisms underlying the effects of nanomolar concentrations of neurosteroids at the NMDA receptor. </p><p>The results show that the neurosteroids pregnenolone sulfate (PS) and pregnanolone sulfate 3α5βS) differently modulate the NMDA receptor, changing the kinetics for the NMDA receptor antagonist ifenprodil, through unique and separate targets at the NR2B subunit. The effects that appear to be temperature independent were further confirmed in a calcium imagining functional assay. A second functional study demonstrated that PS and 3α5βS affect glutamate-stimulated neurite outgrowth in NG108-15 cells. </p><p>Misuse of anabolic androgenic steroids (AAS) has powerful effects on emotional states. Since neurosteroids regulate processes involved in mood it can be hypothesised that AAS can interact with the action of neurosteroids in the brain. However, chronic administration of the AAS nandrolone decanoate did not alter the allosteric effects of PS or 3α5βS at the NMDA receptor, but changed the affinity for PS, 3α5βS and dehydroepiandrosterone sulfate to the sigma 1 receptor. The results also showed that the neurosteroids displace ³H-ifenprodil from the sigma 1 and 2 receptors without directly sharing the binding site for ³H-ifenprodil at the sigma 1 receptor. The decreased affinity for the neurosteroids at the sigma 1 receptor may be involved in the depressive symptoms associated with AAS misuse.</p><p>The NMDA receptor system is deeply involved in neurodegeneration and the NMDA receptor antagonist ifenprodil exert neuroprotective actions. The findings that neurosteroids interact with ifenprodil at the NMDA receptor may be an opportunity to obtain synergistic effects in neuroprotective treatment.</p>

A new group of anabolic androgenic steroid (AAS) users has developed during the last two decades. This group consists primarily of young men interested in improving their physical appearance. Within this group, AAS are sometimes used together with other illicit drugs, alcohol and nicotine. Brutal and violent crimes have been committed under the influence of AAS, possibly because of AAS psychiatric side effects, ranging from increased aggression and psychosis to depression. Unfortunately, the biochemical mechanisms behind these effects are poorly understood. In this thesis we used an animal model to study biochemical and behavioural effects of chronic AAS treatment (15 mg/kg/day of nandrolone decanoate for 14 days). The effect on the endogenous opioid peptides and the expression of immediate-early gene protein Fos in various brain regions were studied using radioimmunoassay and immunohistochemistry, respectively. In addition, we studied AAS effect on voluntary alcohol consumption and defensive behaviours, including aggression. The results show that AAS enhance endogenous opioid activity and Fos expression in brain regions regulating reward, aggression and disinhibitory behaviours. An imbalance between two opioid systems with generally opposing effects, the enkephalins with euphoric and the dynorphins with dysphoric effects, was also found. This implies that AAS alter the ability to maintain a stable state of mind and the response to other drugs of abuse. The AAS pre-treated animals enhanced their alcohol intake, were more aggressive and showed lower fleeing and freezing reaction than the controls. In addition, AAS enhanced amphetamine-induced aggression when the amphetamine was given three weeks after the last AAS injection. The behavioural and biochemical results found in this thesis, support the hypothesis that use of AAS might lead to the development of dependence and may induce changes in the brain leading to disinhibitory behaviours.

The neurosteroids constitute a group of powerful hormones synthesized and acting in the central nervous system. They participate in a number of important central processes, such as memory and learning, mood and neuroprotection. Their effects emerge from rapid interactions with membrane bound receptors, such as the N-methyl-D-aspartate (NMDA) receptor, the gamma-amino-butyric acid receptor and the sigma 1 receptor. The mechanisms of action are separate from classical genomic interactions. The aims of this thesis were to identify and characterize the molecular mechanisms underlying the effects of nanomolar concentrations of neurosteroids at the NMDA receptor. The results show that the neurosteroids pregnenolone sulfate (PS) and pregnanolone sulfate 3α5βS) differently modulate the NMDA receptor, changing the kinetics for the NMDA receptor antagonist ifenprodil, through unique and separate targets at the NR2B subunit. The effects that appear to be temperature independent were further confirmed in a calcium imagining functional assay. A second functional study demonstrated that PS and 3α5βS affect glutamate-stimulated neurite outgrowth in NG108-15 cells. Misuse of anabolic androgenic steroids (AAS) has powerful effects on emotional states. Since neurosteroids regulate processes involved in mood it can be hypothesised that AAS can interact with the action of neurosteroids in the brain. However, chronic administration of the AAS nandrolone decanoate did not alter the allosteric effects of PS or 3α5βS at the NMDA receptor, but changed the affinity for PS, 3α5βS and dehydroepiandrosterone sulfate to the sigma 1 receptor. The results also showed that the neurosteroids displace 3H-ifenprodil from the sigma 1 and 2 receptors without directly sharing the binding site for 3H-ifenprodil at the sigma 1 receptor. The decreased affinity for the neurosteroids at the sigma 1 receptor may be involved in the depressive symptoms associated with AAS misuse. The NMDA receptor system is deeply involved in neurodegeneration and the NMDA receptor antagonist ifenprodil exert neuroprotective actions. The findings that neurosteroids interact with ifenprodil at the NMDA receptor may be an opportunity to obtain synergistic effects in neuroprotective treatment.

Aromatase is an enzyme that facilitates the conversion of androgens to estrogens and may play a role in mood and mental status. The main theme of this thesis is the imaging of brain aromatase by use of the PET technique. The PET tracer for aromatase, 11C-labeled vorozole (VOZ) was developed and evaluated by with in vitro and in vivo methods. In vitro experiments using rat brain showed that VOZ was distributed in the medial amygdala, bed nucleus of the stria terminalis and medial preoptic area, regions of the brain known to be rich in aromatase and the KD value was determined to be 0.60 nM. The in vivo PET study in rhesus monkey brain revealed that VOZ penetrated the blood-brain barrier and accumulated in the amygdala and hypothalamus. Taken together, VOZ is a good PET tracer for in vivo aromatase imaging with high affinity and high sensitivity. This technique was applied to an investigation of brain aromatase under the physiological conditions simulating anabolic-androgenic steroid abuse. A significant increase in VOZ binding by anabolic-androgenic steroids was observed in the bed nucleus of stria terminalis and medial preoptic area in the rat brain. In contrast, no significant change in binding was observed in the medial amygdala. These results indicate that the manner of regulation of aromatase expression might be different in the bed nucleus of stria terminalis and medial preoptic area compared with that in the medial amygdala. The aromatase expression was suggested to be regulated through androgen receptors, as indicated in a study with flutamide treatment. The increased aromatase expression was seen in neurons. The PET study with anabolic steroid-treated rhesus monkeys also showed increased VOZ binding in the hypothalamus but not in the amygdala. The alteration of density of aromatase binding in the hypothalamic area could explain some psychological features of anabolic-androgenic steroid abusers. Novel PET tracers for aromatase were developed and examined. The two newly synthesized 18F-labeled vorozole analogs, [18F]FVOZ and [18F]FVOO, displayed different characteristics. Both tracers showed similar binding pattern as VOZ; however, [18F]FVOO was metabolized very quickly, meaning that this tracer is not suitable as a PET tracer. On the other hand, [18F]FVOZ can be an appropriate PET tracer. The role of aromatase in the human brain has not been clarified yet. To approach this problem by in vivo methods, we have just started PET studies to explore aromatase expression in humans.

Die missbräuchliche Anwendung von anabolen Substanzen erfolgt mit dem Ziel eines verstärkten Muskelaufbaus - im Sport zur Leistungsverbesserung, in der Tierzucht zum Erreichen von Zuchtidealen oder bei der Masttierhaltung zur Produktivitätssteigerung. Bisher wurden Doping- oder Medikationskontrollen zum Nachweis von anabolen Steroidhormonen üblicherweise im Urin bzw. im Blut durchgeführt. Für bestimmte Fragestellungen kann der analysierbare Zeitraum allerdings unzureichend sein oder aber die Untersuchungsmaterialien sind unter praktischen Gegebenheiten nur eingeschränkt verfügbar. Das Sammeln von Urinproben ist beispielsweise bei Zuchthengsten nur mit einem unverhältnismäßig hohen Aufwand realisierbar. Haare stellen in solchen Situationen eine Alternative dar, da sich das Entnahmeverfahren unkompliziert gestaltet und bei einer entsprechenden Haarlänge die eingelagerten Fremdstoffe länger als in Urin- oder Blutproben detektierbar sein sollten. In der vorliegenden Arbeit wurde ein effektiver Langzeitnachweis für insgesamt 11 anabole Substanzen in Pferdehaar-Proben mittels GC-HRMS und GC-MS/MS entwickelt (Nachweisgrenzen zwischen 0,1 und 5,0 pg/mg). Dabei können zum einen körperfremde anabole Wirkstoffe (z. B. Steroidester in Depotpräparaten) und zum anderen körper-eigene Steroide analysiert werden (z. B. Testosteron und Nandrolon beim Hengst). In verschiedenen Applikationsversuchen wurde gezeigt, dass durch eine Haaranalyse der Nachweis bis zu einem Jahr möglich ist. Für die endogene Nandrolonmenge in Schweifproben von unbehandelten Hengsten wurde eine signifikante Altersabhängigkeit festgestellt. Die ermittelten physiologischen Höchstkonzentrationen für Nandrolon betragen zwischen 1,1 pg/mg bei Junghengsten (1-3 Jahre) und 3,1 pg/mg bei Althengsten (11-20 Jahre). Die Bestimmung von Nandrolon in Haarproben erwies sich für die Körungskontrollen bei Junghengsten als ein geeignetes Verfahren zur Detektion einer exogenen Zufuhr. Die Untersuchung von Haaren ist zum Langzeitnachweis als Alternative gegenüber Blut- und Urinanalysen vorzuziehen, auch wenn sich retrospektiv nicht alle Fragen zum Behandlungsablauf präzise klären lassen (z. B. Angaben zur Dosierung oder zum genauen Applikationszeitpunkt). Das neu etablierte Verfahren ist außerdem die Methode der Wahl, wenn die Verfügbarkeit der übrigen Probematerialien eingeschränkt bzw. eine einfache und schnelle Beprobung erforderlich ist. Es wird bereits zur Medikationskontrolle bei Zuchthengsten sowie bei speziellen forensischen Untersuchungen eingesetzt.

Objective: Despite widespread drug testing in sports and warnings about the potential risks of using anabolic-androgenic steroids (AAS), non-medical use is prevalent among athletes, non-athletes, and disturbingly among adolescents. To date, most research has focused on the anabolic properties and short-term health risks of AAS use. In contrast, studies investigating the effects on cognitive function in men using high doses of multiple exogenous steroids are lacking. The primary purpose of this naturalistic study was to examine the effects of non-medical steroid use on sex-related cognitive abilities in male bodybuilders. The secondary purpose of the study was to evaluate the psychological functioning of male bodybuilders who use AASs. Methods: Eight male bodybuilders who used high doses of AASs were matched with bodybuilding and aerobic controls who had never used AASs, according to age, education, and estimated verbal intelligence. AAS use of the bodybuilders appeared similar to reports in the literature of self-administered AASs regimens used by strength athletes. All groups underwent a battery of cognitive tests and self-report psychological inventories, and had serum total testosterone and binding proteins measured immediately after testing. Cognitive measures selected were those that have previously shown sex differences. The study examined four psychological domains: aggression, personality, body image, and eating-disordered attitudes/behaviours. Results: Male bodybuilders who used AASs scored significantly lower than controls on mental rotations and on the WAIS-III Digit-Symbol Coding subtest. There were no other significant group differences on the cognitive tasks. A curvilinear (inverted U) relationship was identified between spatial ability and total testosterone in men who did not use AASs. As there were only a few AAS users in the current study, there was little power to demonstrate a linear or nonlinear relationship. Overall, there were no significant differences between groups on the psychological variables. AAS users exhibited elevated levels of antisocial personality traits, with 38% scoring in the clinically significant range. Bodybuilders reported some body weight concerns, specifically a drive for muscularity combined with a drive for a well-toned body, with no difference between AAS users and bodybuilding controls. Three AAS users and one bodybuilding control exhibited psychological disturbances, as evidenced by elevated scores on multiple psychological measures. Conclusions: The results of this preliminary study provide some evidence that high doses of AASs in men might influence certain aspects of cognition, specifically reducing complex visuospatial skills and perceptual speed. The data also suggests that endogenous testosterone influences spatial ability in healthy men in a curvilinear fashion. Further research with larger samples of AAS users is required to quantify the cognitive effects of non-medical AAS regimens. The study also contributes to the growing literature on the psychological effects of bodybuilding and AAS use. Although many AAS users and bodybuilders might display minimal psychopathology, there is likely a subgroup of individuals who exhibit clinically significant psychological disturbances. Further research is necessary to identity the nature and severity of psychological symptomatology in this population, and effective modes of treatment.

O uso indiscriminado de recursos ergogênicos como os esteróides anabolizantes vêm se tornando um problema crescente em diversos segmentos da população, além do meio atlético, tendo como finalidade a obtenção de melhor desempenho físico ou simplesmente melhor aparência física, porém muitas vezes sem a preocupação com os riscos dos efeitos colaterais promovidos por esta prática. No presente trabalho estudamos os efeitos da associação do uso de doses suprafisiológicas de esteróides anabolizantes e do treinamento físico aeróbio de natação sobre o fluxo sangüíneo coronário de ratos normotensos, verificando a participação da adenosina como um dos possíveis mecanismos de regulação deste fluxo, além de alterações estruturais cardíacas que poderiam influenciar na perfusão sangüínea cardíaca. Ao observarmos somente o efeito do treinamento físico, verificamos que o mesmo foi eficaz em promover adaptações benéficas ao sistema cardiovascular, como a presença de hipertrofia cardíaca fisiológica e melhora no fluxo sangüíneo coronário em repouso, provavelmente mediado por uma maior formação de adenosina circulante e cardíaca. O uso de esteróides anabolizantes associado ao treinamento físico atenuou os efeitos benéficos promovidos pelo treinamento, observando-se a presença de hipertrofia cardíaca acompanhada por redução de débito cardíaco e fluxo sangüíneo coronário, mediado por menor produção de adenosina circulante, além de prejuízo na resposta vasodilatadora à acetilcolina, demonstrando uma provável disfunção endotelial e redução na densidade capilar cardíaca, caracterizando desta forma, um quadro patológico / The abusive use of ergogenic resources as the anabolic steroid became an increasing problem in several segments of the population, beyond the athletical way, searching for better performance or physical appearance, without being worried about the risks of the collateral effects promoted by this practice. In the present work we studied the effects of the use of supraphysiological doses of anabolic steroids associated with aerobic swimming training on the coronary blood flow of normotensive rats, investigating the participation of adenosine as one of the possible mechanisms of blood flow regulation, besides the cardiac structural alterations that could influence the coronary blood perfusion. The effect of the physical training was efficient to promote beneficial adaptations of the cardiovascular system, as the presence of physiological cardiac hypertrophy and improves the coronary blood flow at rest, probably mediated by a higher circulating and cardiac adenosine production. The use of anabolic steroids associated with the swimming training attenuated the beneficial effect promoted by training, being observed the presence of cardiac hypertrophy, followed by reduction of cardiac output and coronary blood flow, mediated by lower circulating adenosine production, besides the impairment of the vasodilator response to the acetylcholine, demonstrating a probable endothelial dysfunction and reduction of the cardiac capillary density, characterizing in this way, a pathological state

O uso hormônios como promotores de crescimento em animais de criação foi proibido em vários países, inclusive no Brasil, desde os anos setenta. Para o controle destes anabolizantes, foi desenvolvida uma enorme variedade de métodos, tanto com propósitos de triagem como de confirmação. Os métodos mais usados são os físico-quimicos, entre eles os cromatográficos, tais como cromatografia em camada delgada, cromatografia a gás acoplada a espectrômetro de massa, cromatografia líquida de alta eficiência com detector UV, e os ensaios imunoquímicos tais como radioimunoensaio ou enzimaimunoensaio. Este trabalho revisa os métodos analíticos mais usados internacionalmente no controle de substâncias anabolizantes em tecidos e urina de animais de criação para consumo humano. Atenção especial é dada ao anabolizante potencialmente cancinogênico, Dietilstilbeltrol (DES) em relação ao seu uso no Brasil e às dificuldades envolvidas em sua determinação. São propostos neste trabalho métodos para detecção do DES em sítios de aplicação, na urina, no tecido muscular e nas vísceras, bem como são apresentados os resultados da aplicação desta metodologia em amostras de animais implantado e de controle e amostras autênticas. / Since the seventies, the use of hormonal anabolics as growth promotors in lifestock fattening has been banned in many countries, including Brazil. For control purposes a wide range of methods have been developed for both large-scale screnning and confirmation. The most used methods are the physical-chemical ones, such as thin-layer chromatography (TLC), gas chromatography-mas spectrometry (CG-MS), high performance liquid cromatography (HF\'LC) with UV detector and immunochemical assay (RIA) or enzime imuno assay (ELISA). This thesis reviews the most used methods for the control of anabolics in tissue as well as in the urine of cattle raised for human consumption. Special attention is given to the potencially calcinogen Diethylstilbestrol, its possible use in Brazil and the difficulties envolved in its determination. Suitable methods for the detection of DES in samples from injection sites, urine, tissue and organs of implanted and control animals, are presented, as well as results obtained from authentic samples.