Existem evidências na literatura que demonstram a existência de células indiferenciadas na capsula adrenal, e que o ACTH poderia estimular estas células. Porém não se sabe quais os genes e vias que desencadeiam esta resposta. Através de animais Cre-Lox induzível por Tamoxifeno, silenciamos o gene Pomc em camundongos adultos e avaliamos o efeito do ACTH nessas células. Foram utilizadas placas de PCR array para análise de genes relacionados com células progenitoras em amostras obtidas pela técnica de rolamento, e validação por PCRq com amostras microdissecadas da zona capsular/subcapsular da adrenal. Após caracterização dos animais com o gene Pomc silenciado e tratamentos com ACTH observamos o aumento da expressão de genes relacionados com as vias Wnt, Igf1 e Notch. Esses dados corroboram evidencias descritas na literatura que mostram a importância dessas vias no desenvolvimento e manutenção do córtex adrenal, e sugerem o envolvimento do ACTH nesses processos que envolvem as células progenitoras do córtex adrenal. / There is evidence in the literature demonstrating the existence of stem cells in the adrenal capsule, and that ACTH could stimulate these cells. However, it remains unknown which genes and pathways that trigger this response. By using a tamoxifen-inducible Cre-Lox mice strain, we knocked-out Pomc gene in adult mice and evaluated the effect of ACTH in these cells. PCR array technique was used to determine the expression level of key genes related to progenitor cells in samples obtained by the technique of \"rolling bearing\". Also, we validated the data by qPCR using samples from microdissected capsular areas of the adrenal gland. After characterization of animal model, the results show that treatment with ACTH increase the expression of genes related to Wnt, Igf1 and Notch pathways. These data corroborate with the literature, reinforcing the importance of these pathways in the development and maintenance of the adrenal cortex, and also suggesting the involvement of ACTH in these processes involving the progenitor cells of the adrenal cortex.

La spectroscopie RMN à haute résolution (HR) en rotation à l'angle magique (Magic AngleSpinning, MAS) permet la caractérisation métabolomique tissulaires à partir de faibles quantités de matériel. Nous avons caractérisé par spectroscopie RMN HRMAS le profil métabolomique du neuroblastome de l'enfant et des principales tumeurs surrénaliennes chez l'adulte. Nos résultats se situent parmi les premières explorations par spectrométrie RMN HRMAS de la surrénale. Concernant le neuroblastome, le profil métabolique tumoral a été comparé à celui de la médullosurrénale normale. La relation entre le phénotype métabolomique et des facteurs pronostiques établis en cl inique a été étudiée. L'évolution cl inique des patients a été enfin corrélée au profil métabolomique. Chez l'adulte, nous nous sommes intéressés à l'étude de l'adénome, du corticosurrénalome et du phéochromocytome. Le métabolome tumoral a été ainsi comparé au profil sécrétoire biologique et génétique tumoral. Nos résultats montrent que la spectroscopie RMN HRMAS est une méthode très prometteuse pour l'étude du profil métabolomique de la surrénale et de ses principales pathologies tumorales. / 1 H-high-resolution magic-angle spinning nuclear magnetic resonance (HRMAS NMR) spectroscopy is one of the most innovative technologies for a global study of metabolism analyzing intact tissue samples. We have assessed the metabolic profile of the normal adrenal and its principal primary tu mors in children and adult. Our study represents one of the first applications of HRMAS NMR spectroscopy to adrenal pathology. Firstly, we have analyzed the pediatric neuroblastoma. The tumoral metabolic profile was compared to that of normal adrenal medulla.Thereafter, the relationship between tumoral metabolomic phenotype and selected established prognostic factors was studied. Patient clinical course was finally correlated to tumoral metabolomic profile. ln adults subjects we have established the metabolic profile of adenoma, adrenal cortical carcinoma and pheochromocytoma. Tumor metabolome was correlated to hormonal secretion and patient's genetic profile. Finally, our findings show that HRMAS NMR spectroscopy is a very promising method for the study of metabolomic profile of tumors originating from the adrenal gland in children and adult subjects.

The work in this thesis consisted of initial experiments designed to elucidate the role of hormones in a hormonal dependent tumor. Various aspects of nucleic acid synthesis in a hormone dependent tumor in the presence (growing) and absence (regressing) of the hormone were studied. The rates of nucleic acid synthesis were studied in whole animals by injecting radioactive formate and allowing the animal to incorporate radioactivity for various periods of time. Nucleic acids were extracted by PAS, phenol procedure and separated on a MAK column. Labelling of all species of nucleic acid was decreased in regressing tumors. In order to determine whether estrogen is acting directly on cells or at some indirect physiological level; the ability of cells from growing and regressing tumor to synthesize nucleic acids in vitro was determined. Results of experiments with these cell suspensions demonstrate that cells from the regressing tumor had a decreased ability to synthesize nucleic acids relative to growing tumor. The rate of DNA synthesis was decreased somewhat more than RNA. In preliminary experiments the activity of DNA dependent DNA polymerase and RNA polymerase from regressing tumor was compared with the same enzyme in growing tumor. The specific activity of both RNA and DNA polymerase was decreased in the regressing tumor. In target tissue like uterus stimulation with estradiol results in an increased rate of synthesis of several species of RNA. In the tumor system used in these preliminary experiments, stimulation with estrogens has a greater effect on the synthesis of DNA than RNA. / Medicine, Faculty of / Biochemistry and Molecular Biology, Department of / Graduate

Made available in DSpace on 2016-05-02T13:53:53Z (GMT). No. of bitstreams: 1 Amarildo Marciano Rosa-dissertacao.pdf: 2611071 bytes, checksum: dc812aaabc78398eb9057cb36f0aeba0 (MD5) Previous issue date: 2013-05-24 / This work was performed in the laboratories of animal reproduction, pharmacology and experimental surgery, university josé do rosário vellano-unifenas, in order to evaluate some aspects related to the morphological changes of cortical adrenal glands and testicles in rats treated with dexamethasone, an anti-inflammatory corticosteroid with immunosuppressive activity in the hypothalamic-pituitary-adrenal.The experiments were conducted to determine the hematological parameters, hormonal and functional morphology of the testicles and adrenal glands of wistar rats (Rattus norvegicus) submitted to dexamethasone at a dose of 40&#956;g/100g weight by 3 days and 7 days of application. The groups were formed by 40 male wistar rats, adult, with average weight of 330g were divided into groups of 10 animals, 10 control and 10 treatment for 3 days and 10 control and 10 treatment for 7 days, where the group treatment received 0.07 ml of dexamethasone intramuscularly and the control group received 0.07 ml of saline intramuscularly. Data were expressed as mean ± standard error of mean (x ± sem), analyzed and compared using the student t-test, with significance level of 5% (p <.05). Were analyzed by the statistical package graphpad instat ® _, version 3.00 (graphpad software inc., san diego, california, usa). Significant differences were found between control and dexamethasone groups in hematological parameters (p <0.05), hormonal (p <0.05) and morphological (p <.05). There was no significant difference in spermiograms. It was concluded that the use of dexamethasone was responsible for the reduction in immune response with a significant lymphopenia, hormonal suppression of cortisol production by the adrenal glands atrophy of adrenal glands, and reduction in weight of the animals, representing the same catabolic effect on body. No interference in the structure of the testicles morphofunctional significantly, nor were there significant changes in the concentration of total testosterone. / Este trabalho foi realizado nos laboratórios de Reprodução Animal, Farmacologia e Cirurgia Experimental da Universidade José do Rosário Vellano -UNIFENAS, com o objetivo de avaliar alguns aspectos relacionados às alterações morfofuncionais testiculares e da cortical da glândula adrenal em ratos tratados com dexametasona, um anti-inflamatório corticosteroide, com atividade imunossupressiva no eixo hipotálamo-hipófise-adrenal.Os experimentos foram realizados para se determinar os parâmetros hematológicos, hormonais e morfofuncionais dos testículos e glândulas adrenais de ratos wistar(Rattus norvegicus) submetidos à dexametasona, na dose de 40µg/100g de peso, por 3 dias e 7 dias de aplicação. Os grupos foram formados por 40 ratos Wistar albinos machos, adultos, com peso médio de 330g, divididos em grupos de 10 animais, sendo 10 controle e 10 tratamento, por 3 dias e 10 controle e 10 tratamento, por 7 dias, onde o grupo tratamento recebeu 0,07mL de solução de dexametasona, via intramuscular e o grupo controle recebeu 0,07ml de solução salina, via intramuscular. Os dados obtidos foram expressos por médias ± erro padrão da média (X±EPM), analisados e comparados usando o Teste t-Student, com significância de 5% ( P<0,05). Foram analisados pelo pacote estatístico GraphPad InStat®_, versão 3.00 (GraphPad Software Inc., San Diego, Califórnia, EUA). Foram encontradas diferenças significativas entre os grupos dexametasona e controle nos parâmetros hematológicos (P<0,05),hormonais(P<0,05)e morfológicos(P<0,05). Não foi evidenciada diferença significativa nos espermogramas. Concluiu-se que a utilização de dexametasona foi responsável pela redução na resposta imune com significativa linfocitopenia, supressão da produção hormonal de cortisol pelas glândulas adrenais, atrofia das glândulas adrenais e pela redução de peso dos animais, representando o efeito catabólico da mesma no organismo. Não houve interferência na estrutura morfofuncional dos testículos de forma significativa, nem houve alteração significativa na concentração detestosteronatotal.

Tema : Audinių prieinkstinių liaukų histopatologinis tyrimas. Tyrimo objektas: X ūkio 6 mėnesių amžiaus audinių prieinkstinių liaukų histologiniai preparatai. Atlikimo vieta: Lietuvos Sveikatos Mokslų Universiteto Veterinarijos Akademijos Patologijos centras. Darbo apimtis: 41 puslapis, naudoti 43 literatūros šaltiniai, 31 paveikslas. Tyrimo tikslas: Įvertinti audinių prieinkstinių liaukų histomorfologinius pokyčius. Tyrimo uždaviniai: 1. Atlikti histologinį ir histocheminį mėginių tyrimus. 2. Įvertinti audinių prieinkstinių liaukų histopatologijas. Tyrimo metodai: Rutininio skrodimo metu buvo paimta 40 audinių prieinkstinių liaukų pataloginė medžiaga (n=40), fiksuota 10 proc. neutraliu formalino tirpalu, atliktas mėginių dažymas hematoksilinu eozinu ir Kongo raudonuoju. Histologinio tyrimo metu įvertintos prieinkstinių liaukų patologijos. Tyrimo duomenų statistinis įvertinimas atliktas, naudojant Microsoft Exel (2010) programą. Paskaičiuotas įvairių patologijų pasireiškimo dažnumas, jų pasiskirstymas žievėje ir šerdyje (proc.). Taip pat įvertinta, kaip pasiskirstė patologijos žievės zonose, jų pasireiškimo dažnumas procentais. Išvados: 1. Atlikus histologinį tyrimą pakitusių audinių antinksčių rasta 92 proc. 2. Atlikus histocheminį prieinkstinių liaukų tyrimą, baltymo amiloido sankaupų prieinkstinėse liaukose nerasta. 3. Didžiausią dalį patologinių pokyčių sudaro plazmocitų infiltracija (70 proc.) ir hemosiderozė (17 proc.). 4. Mažiausią dalį patologinių pokyčių... [toliau žr. visą tekstą] / Theme: Histopathological research of mink adrenal glands Study object: adrenal gland histological samples (preparations) of 6-month-old minks from farm X. Venue of performance: in Pathology Center of the Veterinary Academy of the Lithuanian University of Health Sciences Volume of the paper: the total volume of the work comprises 41,i.e. 31 picture. The aim of the research: to evaluate histomorphologic changes of mink adrenal glands. Tasks of the work: 1. to make histological and histochemical analysis of samples. 2. to evaluate histopathologies of mink adrenal glands. Research methods: during the routine autopsy was taken pathological material of adrenal glands (n = 40) of 40 minks, which was fixed 10 per cent with neutral formalin solution, and samples dyeing with hematoxylin eosin and Congo red were carried out. Statistical evaluation of the survey data carried out using Microsoft Excel (2010) program. Calculated the frequency of the appearance of various pathologies, their distribution in the cortex and core (in percent). Also, has been assessed the distribution of pathology in cortex areas and their frequency of occurrence percentage (in percent). The most important conclusions : 1. An analysis of 40 male minks adrenal glands histological results showed that 92 per cent. analyzed samples demonstrate a change. 2. The largest part of the pathological changes consist of infiltration by plasmocytes (70 percent.) and hemosiderosis (17 percent.). 3. The smallest part of the... [to full text]

Call number: LD2668 .T4 1953 R62 / Master of Science

Polycystic ovary syndrome (PCOS) is a common reproductive, endocrine and metabolic disorder present in women of reproductive age. Despite the widespread prevalence and heritability of PCOS, the heterogeneous and polygenic traits have made the successful identification of candidate genes difficult. Animal models have been developed on the premise that early exposure to sex steroids can programme epigenetic changes that predispose the fetus to the adult features of PCOS. Past research has modelled ovarian dysfunction, endocrine abnormalities and metabolic perturbances in rodent, non-human primate and sheep PCOS models, through the enhanced neonatal or prenatal exposure to the male sex hormone, testosterone. The modelling of PCOS in a large domestic species such as the sheep is advantageous due to similar biological reproductive function as the human. In this regard the sheep has been extensively used to model PCOS by the treatment of pregnant ewes from early to midgestation with androgens such as testosterone propionate (TP). These experiments have demonstrated the fetal programming effects of androgens on offspring that go on to develop PCOS-like characteristics in adulthood. One of the caveats of assessing steroid effects in this way is the effect of the placenta in mediating the transfer of these hormones. TP is an aromatisable androgen and thus some of its effects in the fetus may be attributable to placental by-products such as estrogens. This thesis describes the development and assessment of a novel model of prenatal androgenisation. Two models were compared: the indirect maternal exposure to TP (the current model) and the direct fetal injection of TP. In directly treating the fetus this allowed control over the dose of TP administered and avoidance of secondary effects that androgens may exert in the mother that could be transferred to the fetus. For the maternal model, pregnant Scottish Greyface ewes were administered TP twice weekly from day (d)62-102 of a 147 day gestation. For the fetal model, fetuses were injected twice while the ewe was anaesthetised with graded doses of TP during the same period of treatment as the maternal model. The effects of prenatal androgenisation were assessed in the female fetus shortly after treatment and also in young adult sheep. Fetal ovarian and adrenal steroidogenic gene expression was monitored and found to be altered in response to elevated levels of sex steroids. At d90 the morphology of the developing ovary was not changed by prenatal androgens. In the adult a detailed ovarian and endocrine assessment was undertaken, by examination of ovarian morphology, hormone levels, ovulatory cycles, hypothalamic pituitary ovarian function and follicle steroidogenesis, during the first breeding season. In addition, the metabolic effects of prenatal androgens were monitored by measuring body fat, insulin and glucose homeostasis and liver function. Neither maternal nor fetal prenatal androgenisation during mid-gestation resulted in a perturbed hormonal milieu or polycystic ovaries in young adults. These treatments did however programme a clear ovarian phenotype demonstrated by the increased capacity of follicles to secrete androgens, independently of an abnormal endocrine environment and disordered folliculogenesis. Furthermore, animals that were exposed maternally to TP developed fatty liver and had increased insulin secretion in response to glucose load. A major outcome of this study was the finding that the fetally injected control animals were phenotypically different than the maternal control animals. In fact, some of the reproductive and metabolic features of maternal TP exposure were found in the fetal control group. This unexpected finding has raised the possibility that it is the fetal exposure to stress, that is secondary to elevated maternal androgens, rather than androgens per se that is responsible for at least some of the multitude of anomalies encountered in PCOS.

Thesis (MSc)--Stellenbosch University, 2003. / ENGLISH ABSTRACT: This study describes: 1. The isolation of total RNA and mRNA from Angora goat adrenals. 2. Synthesis and nucleotide sequence alignment of Angora goat CYPI7 cDNA. Two DNA sequences were produced, identifying two CVP 17 alleles in an Angora goat from the Swartland district. 3. The development of a CYPI7 genotype test for Angora goats. 4. Genotyping of Angora goats and Boer goats with the developed genotype test. S. Mapping of the substituted amino acids in the amino terminal of CVP 17 to a specific CYPI7 genotype. 6. Partial synthesis and alignment of Angora goat genomic nucleotide CYPI7 sequences. / AFRIKAANSE OPSOMMING: Hierdie studie beskryf: 1. Die isolering van totale RNA en mRNA van Angorabok byniere. 2. Sintese en nukleotied volgorde oplyning van Angorabok CYP17 eDNA. Twee DNA volgordes is geproduseer, en so is twee CYP17 allele in 'n Angorabok van die Swartland omgewing geïdentifiseer. 3. Die ontwikkeling van 'n CYP17 genotipe toets vir Angorabokke. 4. Genotipering van Angorabokke en Boerbokke met die ontwikkelde genotipe toets. 5. Korrelering van die omgeruilde aminosure in die aminoterminaal van CYPl7 met 'n spesifieke genotipe. 6. Gedeeltelike sintese en oplyning van Angorabok genomiese CYPl7 nukleotied volgordes.

The aim of this thesis is to in various ways explore protein expression in human normal tissue and in cancer and to apply that knowledge in biomarker discovery. In Paper I the prognostic significance of RNA-binding motif protein 3 (RBM3) is explored in malignant melanoma. To further evaluate the prognostic significance of RBM3 expression was assessed in 226 incident cases of malignant melanoma from the prospective populationbased cohort study Malmö Diet and Cancer Study using tissue microarray technique (TMA). RBM3 was shown to be down regulated in metastatic melanoma and high nuclear expression in the primary tumor was an independent marker of prolonged over all survival. As a tool to facilitate clinical biomarker studies the Human Protein Atlas has created a tissue dictionary as an introduction to human histology and histopathology. In Paper II this work is introduced. A cancer diagnosis can be a complex process with difficulties of establishing tumor type in localized disease or organ of origin in generalized disease. Immunohistochemically assisted diagnosis of cancer is common practice among pathologists where its application combined with known protein expression profiles of different cancer types, can strengthen or help dismiss a suspected diagnosis. In Paper III the diagnostic performance of 27 commonly used antibodies are tested in a predominantly metastatic, multicancer cohort using TMA technique. Overall these 27 diagnostic markers showed a low sensitivity and specificity for its intended use, highlighting the need for novel, more specific markers. Breast, ovarian, endometrial and ovarian cancers affect predominantly women. Differential diagnostics between these cancer types can be challenging. In Paper IV an algorithm, based on six different IHC markers, to differentiate between these cancer types is presented. A new diagnostic marker for breast cancer, namely ZAG is also introduced. In Paper V the transcriptomic landscape of the adrenal gland is explored by combining a transcriptomic approach with a immunohistochemistry based proteomic approach. In the adrenal gland we were able to detect 253 genes with an elevated pattern of expression in the adrenal gland, as compared to 31 other normal human tissue types analyzed. This combination of a transcriptomic and immunohistochemical approach provides a foundation for a deeper understanding of the adrenal glands function and physiology.

Memoria para optar al Título Profesional de Médico Veterinario / El objetivo del presente estudio fue relacionar el diagnóstico ecográfico de atrofia/hipoplasia adrenal con el diagnóstico confirmatorio de la enfermedad de Addison vía prueba de estimulación con ACTH, o bien, relación Sodio/Potasio (Na:K) plasmático. Para esto se buscó perros con diagnóstico ecográfico de atrofia/hipoplasia adrenal y, posteriormente, se seleccionó a aquellos que fueron sometidos a una prueba de estimulación con ACTH o que tuvieron una medición de electrolitos (Na, K). Como resultado se encontró que no existía asociación entre el diagnóstico de la enfermedad de Addison y el diagnóstico ecográfico de atrofia/hipoplasia adrenal. Sin embargo, se observó que las glándulas de los perros diagnosticados con enfermedad de Addison fueron significativamente más pequeñas en su diámetro que las de perros sanos, aún cuando en el estudio sólo se incluyó a perros con glándulas atróficas/hipoplásicas. Esto confirmaría los resultados obtenidos en estudios previos (Wenger et al., 2010; Hoerauf y Reush, 1999) que indican que existe una diferencia significativa en el grosor de las glándulas adrenales de perros con enfermedad de Addison en comparación con las glándulas de perros sanos