Introdução: Alterações nos eixos Hipotálamo-Hipófise-Adrenal (HHA) e Hipotálamo-Hipófise-Tireóide (HHT) estão associados a depressão. Objetivo: Avaliar a associação entre depressão e alterações nos eixos HHA, HHT e o estresse precoce (EP) em deprimidos. Metodologia: Foram avaliados 52 deprimidos e 52 voluntários com idade entre 18 e 45 anos. O diagnóstico de depressão foi baseado no DSM-IV e MINI. A gravidade da depressão foi avaliada pela HAM-D-17 e pelo IDB. Foram aplicados o CTQ buscando avaliar eventos estressantes na infância, além de questionário sócio-demográfico e clínico. Voluntários foram pareados segundo sexo, idade, IMC e submetidos aos mesmos questionários. Foram realizadas dosagem de TSH, T4 livre, anticorpos anti TPO e ATG, Cortisol plasmático, ACTH, DHEA-s, lipidograma, glicemia de jejum e cortisol salivar em 05 tempos. Resultados: Todos os deprimidos apresentaram história de EP. Houve concentrações maiores de TSH, anti TPO, anti TG, cortisol plasmático e salivar, ACTH, colesterol total, LDL, VLDL, triglicérides, AUC 8-23 e AUC0-30-60, além de concentrações menores de DHEA-s e HDL em casos do que controles, considerando a primeira e segunda coleta de dados e conforme a amostra estudada. Houve correlações entre as variáveis estudadas. Conclusão: O EP foi um dos fatores de risco para depressão. Achados desse estudo confirmam a literatura quando se compara deprimidos com controles e se relaciona depressão com os eixos HHT, HHA e o EP. Esse foi o primeiro estudo em que se comparou todas as variáveis supracitadas em pessoas CEP e SEP, gerando resultados positivos, além de revelar diferenças de gênero conforme o resultado. / Introduction: Changes in the axis hypothalamus-pituitary-adrenal (HPA) and hypothalamic-pituitary-thyroid (HPT) are associated with depression. Objective: Evaluating the association among depression and changes in the HPA, HPT axes and early stress (ES) in depressed. Methodology: A total of 52 depressed and 52 volunteers aged between 18 and 45 years were evaluated. Depression diagnosis was based on DSM-IV and MINI. The severity of depression was assessed by HAM-D-17 and BDI. The CTQ were applied for assessing stressful events in childhood, as well as socio-demographic and clinical questionnaire. Volunteers were paired by gender, age, BMI, and underwent the same questionnaires. TSH measurement was performed, free T4, anti TPO and ATG antibodies, plasma cortisol, ACTH, DHEA-s, lipid profile, fasting glucose and salivary cortisol in 05 times. Results: All depressed had history of ES. There were higher concentrations of TSH, anti TPO, anti TG, plasma and salivary cortisol, ACTH, total cholesterol, LDL, VLDL, triglycerides, AUC 8-23 and AUC0-30-60 and lower concentrations of DHEA-s and HDL in cases than controls, taking into account the first and second data collection and according to the studied sample. There were correlations between variables. Conclusion: The ES was one of the risk factors for depression. Findings of this study confirm the literature when compared depressed with controls and associated depression with HPT, HPA axes and ES. This was the first study in which we compared all the above variables in people WES and WOES, generating positive results and revealed gender differences according to result.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2008. / Härtill 4 uppsatser.

Diss. (sammanfattning) Linköping : Linköpings universitet, 2010. / Härtill 5 uppsatser.

As sirtuínas 1-7 (SIRT) constituem uma família altamente conservada de desacetilases de histonas que, de modo geral, participam da regulação da longevidade em diversos organismos, modulando a resposta celular frente ao stress oxidativo e promovendo mecanismo de reparo de DNA, parada do ciclo celular, estabilidade telomérica, senescência e apoptose celulares. O envolvimento das SIRTs no processo tumorigênico tem sido bastante investigado, contudo ainda não existe descrição do estudo desses genes nos adenomas hipofisários. O objetivo desse estudo foi avaliar a expressão gênica das SIRT1-7 nos somatotropinomas e adenomas hipofisários clinicamente não funcionantes (ACNF) e sua relação com o tamanho e a invasividade do tumor. A expressão das sirtuínas foi ainda correlacionada à expressão dos marcadores de senescência CDKN1A (p21) e CDKN2A (p16) e do proto-oncogene PTTG (pituitary tumor transforming gene). Foram selecionados 68 pacientes, 37 somatotropinomas e 31 portadores de ACNF. Desses casos, 33 apresentavam tumores invasivos e 35 eram não invasivos. A quantificação do RNAm das SIRT1-7, CDKN1A, CDKN2A e PTTG foi realizada nas amostras tumorais pela técnica de PCR em tempo real utilizando o método de quantificação relativa 2-??Ct. A hiperexpressão da SIRT1 foi observada em 86,5% dos somatotropinomas versus 41,9% dos ACNF (P < 0.01), não sendo observada perda de expressão desse gene. A SIRT3 foi mais hipoexpressa nos ACNF em relação aos somatotropinomas (77,4% e 40,5%, respectivamente; P < 0.01). A SIRT4 foi hipo e hiperexpressa, respectivamente, em 45,2% e 12,9% dos ACNF e 16,2% e 24,3% dos somatotropinomas (P=0.03). A hipoexpressão da SIRT7 também foi maior nos ACNF (67,7%) versus somatotropinomas (32,4%; P=0.01) e, para ambos os subtipos, o percentual de casos apresentando hiperexpressão desse RNAm foi baixo. O padrão de expressão das SIRT2 e 5 não diferiu entre os subtipos tumorais e não se mostrou alterado em relação ao pool de hipófises normais. Não foi observada diferença estatisticamente significante na expressão dos genes das sirtuínas entre os grupos de tumores invasivos e não invasivos. Contudo, a expressão das SIRT1 e 3 foi relacionada ao tamanho tumoral; nos casos com hiperexpressão da SIRT1 a média do maior diâmetro tumoral foi 2.4 ± 1.1 enquanto nos pacientes com expressão normal foi de 3.3 ± 1.3 (P < 0.01). Já os casos com perda de expressão da SIRT3 apresentaram tumores maiores (3.1 ± 1.2) em relação aos casos com expressão normal (2.2 ± 1.1; P < 0.01). A expressão de todas as SIRTs apresentou correlação positiva moderada (SIRT1-5,7) ou forte (SIRT6) com a expressão do CDKN1A. Uma correlação positiva foi observada também em relação a expressão do CDKN2A. Contudo, essa foi fraca e presente apenas para as SIRTs 3-5. Em relação ao PTTG, foi observado apenas uma fraca correlação com a expressão da SIRT1 e SIRT3. Em conclusão, esses resultados sugerem que a hiperexpressão de SIRT1 e a hipoexpressão das SIRTs 3, 4 e 7 podem estar relacionadas ao processo tumorigênico nos somatotropinomas e ACNFs, respectivamente e, em especial as SIRT1 e 3, ao controle da proliferação celular nesses adenomas / Sirtuins 1-7 (SIRT) are a highly conserved family of histone deacetylases. In general, these proteins are involved in the regulation of longevity in several organisms, modulating the cellular response to oxidative stress. SIRTs can also regulate DNA repair, telomeric stability, cell senescence and apoptosis. Due to their functions, there is a growing interest in the role of sirtuins in tumorigenesis. However, these genes were not investigated in pituitary tumors so far. In this study, SIRT1-7 gene expression was evaluated in somatotropinomas and nonfunctioning pituitary adenomas (NFPA) and related to tumor size and invasiveness. SIRT1-7 expression was also correlated with cellular senescence markers CDKN1A (p21) e CDKN2A (p16) and with the proto-oncogene PTTG (pituitary tumor transforming gene). Sixty-eight patients were selected, 37 with somatotropinomas and 31 with NFPA. Tumor invasion was observed in 33 patients. SIRT1-7, CDKN1A, CDKN2A and PTTG mRNA levels was evaluated from pituitary tumor samples by the real-time PCR using 2-??Ct relative quantification. Pronounced differences in SIRT1, 3, 4 and 7 expressions were identified between somatotropinomas and NFPA. Overexpression of SIRT1 was observed in 86.5% of somatotropinomas versus 41.9% of NFPA (P < 0.01) whereas underexpression was not detected. SIRT3 was more underexpressed in NFPA than somatotropinomas (77.4% and 40.5%, respectively, P < 0.01). SIRT4 was under and overexpressed, respectively, in 45.2% and 12.9% of NFPA and 16.2% and 24.3% of somatotropinomas (P=0.03). SIRT7 underexpression was also higher in NFPAs (67.7%) versus somatotropinomas (32.4%; P=0.01) with few cases showing overexpression. SIRT2 and 5 expressions did not differ between tumors subtypes and was not altered in relation to the normal pituitary gland pool. No statistically significant difference was observed in the expression of these genes between invasive and non-invasive tumor groups. However, SIRT1 and 3 expressions were related to tumor size. Mean of the largest tumor diameter was 2.4 ± 1.1 and 3.3 ± 1.3 (P < 0.01) in adenomas with SIRT1 over- and normal expression, respectively. On the other hand, cases with SIRT3 underepression exhibited larger tumors (3.1 ± 1.2) compared to cases with SIRT3 normal expression (2.2 ± 1.1, P < 0.01). Moderated (SIRT1-5.7) or strong (SIRT6) positive correlation was observed between sirtuins and CDKN1A expression. A weak correlation was observed with respect to CDKN2A expression and SIRTs 3-5. Regarding PTTG mRNA, only a weak correlation with SIRT1 and SIRT3 expression was observed. In conclusion, these results suggest that overexpression of SIRT1 and underexpression of SIRTs 3, 4 and 7 could be related to the tumorigenic process in somatotropinomas and NFPAs, respectively. SIRT1 and 3 could also play a role in control of pituitary adenomas cell proliferation

Submitted by Erika Demachki (erikademachki@gmail.com) on 2018-02-19T18:29:22Z No. of bitstreams: 2 Dissertação - Edmêe Aparecida Fleury Curado - 2017.pdf: 2300652 bytes, checksum: ae7a412a9c9e2fff2ee8706032b1b995 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-02-20T11:03:42Z (GMT) No. of bitstreams: 2 Dissertação - Edmêe Aparecida Fleury Curado - 2017.pdf: 2300652 bytes, checksum: ae7a412a9c9e2fff2ee8706032b1b995 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-02-20T11:03:42Z (GMT). No. of bitstreams: 2 Dissertação - Edmêe Aparecida Fleury Curado - 2017.pdf: 2300652 bytes, checksum: ae7a412a9c9e2fff2ee8706032b1b995 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-10-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Due to the current trade and technology used in equine breeding, there is a search for effective reproductive methods, seeking to optimize the resources provided by the animal and the environment. Hormonal mare characteristics already known and used with a positive reproductive effect, such as the use of equine pituitary extract (EPE) to induce ovulation, may be better employed if we eliminate the heterogenicity of the samples at the moment of collection. This experiment was conducted with the objective of inducing estrus and ovulation in mares at the end of the mating season, with the use of EPE collected separately at different times of the year - summer and winter. The protocols were 14 cross bred mares, of which 7 were submitted to treatment with EPE collected in the summer and the others with EPE collected in the winter. All of these animals were initially treated as a control group, that is, being monitored daily for ovarian, uterine and behavioral changes from the start of the breending season. From the beginning of March 2017 (03/03/2017), these animals were randomly assigned to one of the treatments, which consisted of an intramuscular application of 125 IU EPE every 12 hours, until one or more more follicles with 34/35 mm, when each one received a single intravenous EPE application containing 500 IU to induce ovulation. The only difference between the groups was the EPE collection season. The results obtained in this experiment allowed to conclude that the reduction of the light incidence after the summer solstice reduces the diameter of the ovulatory follicle. In addition, EPE-summer was more effective when used with the objective of maintaining follicular growth in mares during the end of the breeding season, resulting in larger ovulatory follicles. / Devido ao atual comércio e tecnologia empregada na criação de equinos, existe procura por métodos reprodutivos efetivos, buscando a otimização dos recursos proporcionados pelo animal e pelo ambiente. Caracteristicas hormonais da égua já conhecidas e usadas com efeito reprodutivo positivo, como o uso do extrato de pituitária equina (EPE) para induzir ovulação, pode ser melhor empregado se eliminarmos a heterogenicidade das amostras no momento da coleta. Este experimento foi conduzido com o objetivo de induzir o estro e a ovulação em éguas no final da estação de monta, com o uso de EPE coletado separadamente em diferentes épocas do ano - verão e inverno. Foram submetidas aos protocolos, 14 éguas sem raça definida, sendo 7 delas submetidas ao tratamento com EPE coletado no verão e as outras com EPE coletado no inverno. Todos estes animais foram inicialmente tratados como grupo controle, isto é, sendo monitoradas diariamente quanto as modificações ovarianas, uterinas e comportamentais, desde o início da Estação de Monta. A partir do início de março de 2017 (03/03/2017), estes animais foram randomicamente dividos para um dos tratamentos, que consistiram em uma aplicação por via intramuscular de 125 UI de EPE a cada 12 horas, até a verificação de um ou mais folículos com 34/35 mm, quando então cada uma recebia uma única aplicação de EPE via endovenosa contendo 500 UI, para induzir a ovulação, sendo a única diferença entre os grupos a época de coleta do EPE. Os resultados obtidos neste experimento permitiram concluir que a redução da incidência luminosa após o solstício de verão reduz o diâmetro do folículo ovulatório. Além de que o EPE-verão foi mais eficaz quando utilizado com o objetivo de manter o crescimento folicular em éguas durante o final da estação reprodutiva, resultando em folículos ovulatórios maiores.

Der GLI2-Transkriptionsfaktor ist eines der Haupt Effektor-Proteine des Sonic Hedgehog (SHH)-Signalweges und hat vermutlich eine Schlüsselfunktion in der Entwicklung der Hypophyse. Genomische GLI2-Veränderungen welche zu abgeschnittenen Proteinen führten, wurden beschrieben als Ursache für Holoprosenzephalie (HPE) oder HPE-ähnliche Veränderungen, teilweise in Verbindung mit einer Hypophysenunterfunktion. Ziel dieser Arbeit war die Ermittlung der Frequenz von GLI2-Mutationen in Patienten mit multiplem hypophysärem Hormonausfall (multiple pituitary hormone deficiency, MPHD) und eine funktionelle Untersuchung der gefunden Mutationen mittels Transkriptionsaktivitäts-Untersuchungen durch funktionelle Luciferase assays. Hierfür wählten wir Teilnehmer der GeNeSIS (Genetics and Neuroendocrinology of Short Stature International Study)-Studie. Patienten bei denen bereits Mutationen eines der etablierten Gene für MPHD nachgewiesen wurde, wurden ausgeschlossen. Insgesamt haben wir 168 Patienten mit MPHD untersucht. Bei allen Patienten waren mindestens ein GH- und ein TSH-Mangel dokumentiert, Auffälligkeiten in der zentralen Bildgebung mittels cMRT wurden bei 96 Patienten angegeben. In fünf Studienteilnehmern wurden vier verschiedene heterozygote missense Varianten nachgewiesen, hiervon wurden zwei bislang noch nicht in der Literatur beschrieben. Eine Variante, pR516P, führte in den in-vitro Experimenten zu einem kompletten Verlust der Proteinaktivität. Zusätzlich zu einem Wachstumshormonmangel hatte der Träger dieser Mutation einen Mangel an TSH und der Gonadotropine, sowie einen nichtdeszendierten Hypophysenhinterlappen und eine Polydaktylie, aber keine ersichtlichen Mittelliniendefekte. Anhand der funktionellen Untersuchung konnten wir erstmalig nachweisen, dass ein heterozygoter Aminosäuren-Austausch im GLI2-Protein zu einer möglichen Funktionseinschränkung der Transkriptionsaktivität führen kann und somit die Ursache für MPHD mit milden extrahypophysären Auffälligkeiten sein könnte. Der Phänotyp von GLI2-Mutationen ist variabel und die Penetranz ist unvollständig. GLI2-Mutationen sind assoziiert mit einer Hypoplasie des Hypophysenvorderlappens und treten gehäuft mit einem ektopen Hypophysenhinterlappen auf.:1. Bibliographische Beschreibung S. 4 2. Abkürzungen (alphabetisch sortiert) S. 6 3. Hintergrund S. 8 3.1 Minderwuchs S. 8 3.2 GeNeSIS-Programm S. 10 3.3 Entwicklung, Aufbau und Funktion der Hypophyse S. 11 3.4 Molekularbiologie der Hypophysenentwicklung S. 12 3.4.1 Signalmoleküle, die die Stratifizierung der Rathke-Tasche und die Festlegung der hypophysären Zelllinien bestimmen S. 12 3.4.2 Transkriptionsfaktoren, welche die frühen Phasen der Anlage kontrollieren S. 13 3.5 Hedgehog-Gen Familie S. 15 3.5.1 SHH-Prozessierung und Freisetzung S. 16 3.5.2 SHH-Rezeptorbindung S. 17 3.5.3 SHH-Signaltransduktion S. 17 3.6 GLI-Proteine S. 18 3.6.1 GLI2 S. 19 3.6.2 GLI2-Mutationen bei Menschen S. 20 3.7 Rationale für die Promotionsarbeit S. 20 4. Publikation S. 22 4.1. Druckversion S. 22 4.2 Supplemental Material S. 32 5. Zusammenfassung und Interpretation S. 40 5.1 Screening S. 41 5.2 GLI2-Varianten S. 41 5.3 Experimentelle Untersuchungen S. 42 5.4 Interpretation und Diskussion S. 43 6. Referenzen S. 45 7. Anlagen S. 54 7.1 Erklärung über die eigenständige Abfassung der Arbeit S. 54 7.2 Lebenslauf S. 55 7.3 Publikationen und Auszeichnungen S. 56 7.3.1 Publikationen S. 56 7.3.1 Auszeichnungen S. 56 7.5 Danksagung S. 57

published_or_final_version / abstract / Zoology / Doctoral / Doctor of Philosophy

The purpose of these studies was to determine whether the inhibitory effects of short photoperiod exposure on prolactin cell activity in male Syrian hamsters and/or the inhibitory effects of melatonin treatment on the growth and activity of diethylstilbestrol- (DES) induced prolactinomas in Fisher 344 (F344) rats were possibly mediated through alterations in dopaminergic regulatory mechanisms. In both the hamster and the rat, changes in hypothalamic dopamine neuronal activity and changes in pituitary responsiveness to dopamine have been suggested as possible mechanisms in the prolactin-inhibitory effects of light deprivation or melatonin administration. The present studies in the male Syrian hamster addressed two issues. First, it was of interest to determine if anterior pituitaries of long photoperiod-exposed male hamsters possess dopamine receptors, which are presumably necessary for responsiveness to dopamine. This was accomplished by analysis of ³H-spiperone binding to anterior pituitary membranes. Second, possible changes in pituitary sensitivity to dopamine were assessed by comparison of dose response curves for the inhibition by dopamine of prolactin release from hemipituitaries incubated in vitro from both long and short photoperiod-exposed animals over a series of time points from three to fifteen weeks. In the second series of experiments, adult female F344 rats received daily injection of melatonin or saline vehicle. After two weeks, half of the animals were sacrificed for analysis of ³H-spiperone binding to anterior pituitary membranes, measurement of hypothalamic dopamine turnover and analysis of in vitro pituitary sensitivity to dopamine. The remaining animals received subcutaneous implants containing DES and injections were continued on the same schedule until sacrifice four weeks later for measurement of the same parameters. In both the hamster and rat models, treatments exerted profound inhibitory effects on indices of prolactin cell activity. However, these studies provide no evidence for the involvement of altered dopaminergic regulation in the production of such effects. Neither pituitary sensitivity to dopamine in vitro nor hypothalamic dopamine neuronal activity was enhanced by short photoperiod exposure or melatonin treatment. Prolactin-inhibitory effects of these treatments appear to be mediated through as yet unidentified dopamine-independent mechanisms.