Adenomas hipofisários são tumores benignos, de crescimento lento, originados no interior da sela túrcica e constituem de 10% a 15% dos tumores intracranianos, Os adenomas clinicamente não funcionantes (ACNF), correspondem aproximadamente um terço dos adenomas em geral. Por não apresentarem síndrome clínica hormonal são geralmente diagnosticados devido a sintomas neurológicos ou oftalmológicos, como macroadenomas, com grandes dimensões, invasão de estruturas circunvizinhas e hipopituitarismo. A cirurgia é o tratamento de escolha para estes tumores e apesar de ser eficaz na resolução do quadro compressivo, a possibilidade de cura cirúrgica é reduzida principalmente em tumores invasivos. Seu acompanhamento pós-operatório é efetuado por exame de imagem, preferencialmente ressonância magnética, devido à indisponibilidade de marcadores séricos. Nesta pesquisa avaliou-se a relação da expressão dos genes relacionados à apoptose, Bcl-2, Bax e Caspase-3 e sua relação com o comportamento dos ACNF. Na Divisão de Neurocirurgia Funcional do Instituto de Psiquiatria do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo foram operados 119 doentes com tumores hipofisários, de 28/05/08 à 07/04/09, 50 deles com ACNF, 30 deles foram estudados. A ressonância magnética da região selar pré-operatória possibilitou a medida dos três maiores diâmetros do tumor, ou seja, antero-posterior (AP), crânio-caudal (CC), látero-lateral (LL) e avaliar a invasão do seios cavernoso e esfenoidal. O tamanho dos tumores foi avaliado pela soma dos três diâmetros, pelo maior diâmetro e pelo cálculo do volume, efetuado pela fórmula AP x CC x LL x 0,5. No intraoperatório foram avaliados, a consistência e invasão tumoral. A análise histológica por hematoxilina-eosina, foi efetuada em todos os tumores, assim como a análise imuno-histoquímica (IHQ) dos hormônios hipofisários, Ki-67, p53 e Bcl-2. Foi realizada a análise molecular dos genes Bcl-2, Bax e Caspase-3 por RT-PCR. Dados demográficos: 17 do sexo masculino, 13 do sexo feminino, mediana da idade foi de 54,5 anos e mediana da duração dos sintomas de 31 meses. Todos apresentavam macroadenoma, 87% deles com perda visual, 53% com cefaléia, 17% com outras alterações neurológicas e um assintomático diagnosticado incidentalmente. Avaliação hormonal, disponível em 26 doentes, confirmou deficiência em 92% deles, com mais de dois eixos acometidos em 50% dos casos. A mediana do volume dos tumores foi de 11,6 cm3, do maior diâmetro de 3,8cm e da soma dos três diâmetros de 8,6cm, observou estreita correlação significativa estatisticamente entre as três medidas. Quarenta porcento dos tumores eram gigantes (diâmetro maior ou igual a 4 cm). Consistência amolecida e invasão tumoral foram observadas em 87% e 67% dos tumores, respectivamente. Todos doentes foram operados pela via transesfenoidal, exceto um operado por craniotomia pterional. Complicações cirúrgicas ocorreram em cinco pacientes, três com fistula liquórica, dois com meningite e dois óbitos. A análise histológica confirmou o diagnóstico de adenoma hipofisário em todos os casos. A IHQ foi negativa para todos hormônios em 18 e positiva em 12 tumores (TSH, FSH, LH, GH ou ACTH). A IHQ para proteína P-53 foi negativa em todos os casos. A IHQ para KI-67 revelou ausência da proteína em 11, positividade em menos de 3% das células em 15 e em mais de 3% em 4 tumores. A IHQ para Bcl-2 foi positiva em apenas três pacientes. A análise molecular dos genes Bcl-2, Bax e Caspase-3 revelou expressão muito inferior nos tumores em relação à observada para um pool de hipófise normal. Observou-se correlação positiva estatisticamente significante entre os três genes porém não foi observada correlação entre os níveis destes três genes e nenhum fator de prognóstico tumoral estudado, quais sejam, idade, positividade para hormônios na IHQ, tamanho ou invasão tumoral / Pituitary adenomas are benign, slow-growing tumors that arise in the sella turcica and account for 10% to 15% of all intracranial tumors. Non-functioning pituitary adenomas (NFPA) account for around one third of all pituitary adenomas. NFPA do not clinically present as hormonal syndromes and are generally diagnosed as macroadenomas due to marked neurological and ophthalmologic symptoms and invasion of surrounding structures, beside hypopituitarism. Surgery is the gold standard to treat these tumors. It effectively relieves compressive symptoms but cure is uncommon. Despite benign in nature, NFPA usually show aggressive behavior. There are no hormonal markers and the follow-up usually is made only by magnetic resonance imaging. Apoptosis-related genes, Bcl-2, Bax, and caspase-3, were here studied in NFPA to assess their role as potential markers of tumor behavior. Out of 119 patients with pituitary adenomas treated by surgery, 30 patients (17 men, 13 women, median age 54.5 years old) harboring NFPA who underwent surgery in the Department of Functional Neurosurgery at Hospital das Clínicas Psychiatric Institute, University of S. Paulo Medical School, from August 2008 to July 2009, were studied. Information on gender, age, pituitary function, symptoms and their length was collected. Tumor dimensions were measured using magnetic resonance imaging of the sella turcica. The tumor volume was calculated by the following equation: anterior-posterior diameter x cranial-caudal diameter x lateral-lateral diameter x 0.5. Intra-operative information such as tumor invasion and consistence was recorded. Histological examination by hematoxylin-eosin staining and immunohistochemistry analysis of pituitary hormones, Ki-67, p53, and Bcl-2 were performed. The molecular analysis of Bcl-2, Bax, and caspase-3 genes was performed by real-time polymerase chain reaction (RT-PCR) in all tumor specimens collected during surgery and compared to a poll of normal pituitary gland. All patients had macroadenomas diagnosed due to visual loss (87%), headache (53%) and other neurological symptoms (17%) and one case was incidentally found. Hormonal deficits were seen in 92% of 26 cases; more than two axes were involved in half of these patients. There was found good correlation between tumor volume, largest diameter and the sum of the 3 diameters, and tumor volume was used to assess the correlations with other parameters. The median volume was 11.6 cm3. Giant tumors (4 cm) were diagnosed in 40% of the patients. Soft tumors and tumor invasion were observed in 87% and 67% of cases, respectively. A transsphenoidal approach was used in all patients, except one who had pterional craniotomy. Five patients presented post-operative complications: three had CSF leakage, two meningitis and two died. The histological examination confirmed pituitary adenoma in all cases, 18 of them were null cell and 12 showed a positive immunohistochemistry analysis for one or more hormones, mainly TSH. Immunohistochemistry analysis results for p-53 was negative in all cases; for Ki-67 was negative in 11, positive in less than 3% of the cells in 15 and positive in more than 3% of the cells in 4 cases; and for Bcl-2 was positive only in three patients. Bcl-2, Bax and caspase-3 molecular analysis revealed very low expression compared to normal pituitary values. There was found a positive correlation between these three genes but no correlation between them and age, tumor volume or invasion. The Bcl-2, Bax, and caspase-3 gene analysis by RT-PCR in NFPA did not evidence their potential as markers of tumor behavior

Background: Growth hormone (GH) has been used to treat children with GH deficiency (GHD) since 1966. Aims: Using a combined retrospective and cross-sectional approach, we explored the long-term outcomes of patients with GHD, analysed factors influencing therapeutic response, determined persistence into adulthood, investigated pituitary morphology, and screened for mutations in causative genes. Methods: The files of 96 GH-deficient children were reviewed. In a subset of 50 patients, re-assessment in adulthood was performed, including GHRH-arginine testing, pituitary magnetic resonance imaging (MRI), and mutational screening for the growth hormone-1 gene (GH1) and the GHRH receptor gene (GHRHR) in isolated GHD (IGHD), and HESX1 , PROP1 , POU1F1 , LHX3 , LHX4 , and GLI2 in multiple pituitary hormone deficiency (MPHD) patients. Results: GH was started at a height SDS of –3.2 ± 1.4 in IGHD patients and of –4.1 ± 2.1 in MPHD patients. Relative height gain was 0.3 SDS/year, absolute gain 1.6 SDS, and 1.2/2.6 SDS in IGHD/MPHD, respectively. Mid-parental target height was reached in 77%. Initial height SDS, bone age retardation and duration of GH replacement were correlated with height SDS gain. GHD persisted into adulthood in 19 and 89% of subjects with IGHD and MPHD, respectively. In 1/42 IGHD patients a GH1 mutation was detected; PROP1 mutations were found in 3/7 MPHD subjects. Anterior pituitary hypoplasia, combined with posterior pituitary ectopy and pituitary stalk invisibility on MRI, was an exclusive finding in MPHD patients. Conclusions: GH replacement successfully corrects the growth deficit in children with GHD. While the genetic aetiology remains undefined in most cases of IGHD, PROP1 mutations constitute a major cause for MPHD. Persistence of GHD into adulthood is related to abnormal pituitary morphology.

Indiana University-Purdue University Indianapolis (IUPUI) / The LHX3 transcription factor is essential for pituitary gland and nervous system development in mammals. In humans, mutations in the LHX3 gene underlie combined pituitary hormone deficiency (CPHD) disease featuring deficits in anterior pituitary hormones and defects in the nervous system. The mechanisms that control temporal and spatial expression of the LHX3 gene are poorly understood. The proximal promoters of the human LHX3 gene are insufficient to guide expression in vivo and downstream elements including a conserved 7.9 kilobase (kb) enhancer region appear to play a role in tissue-specific expression in the pituitary and nervous system. In this study, I characterized the activity of this downstream enhancer region in regulating gene expression at the cellular level during development. Human LHX3 enhancer-driven Cre reporter transgenic mice were generated to facilitate studies of enhancer actions. The downstream LHX3 enhancer primarily guides gene transcription in αGSU-expressing cells secreting the TSHβ, LHβ or FSHβ hormones and expressing the GATA2 and SF1 transcription factors. In the developing nervous system, the enhancer serves as a targeting module for expression specifically in V2a interneurons. These results demonstrate that the downstream LHX3 enhancer is important in specific endocrine and neural cell types but also indicate that additional regulatory elements are likely involved in LHX3 gene expression in other cell types. Further, these studies demonstrate significant gonadotrope cell heterogeneity during pituitary development, providing insights into the cellular physiology of this key reproductive regulatory cell. The human LHX3 enhancer-driven Cre reporter transgenic mice provide a valuable tool for further developmental studies of cell determination and differentiation in the pituitary and nervous system. Furthermore understanding the regulation of human LHX3 gene will help develop tools to better diagnose and treat pituitary CPHD disease.

Die vorliegende Arbeit diente vor allem der Untersuchung, ob es im Gehirn vergleichbare Lymphabflusswege wie im übrigen Körper gibt und falls dies zutrifft, wo diese Abflusswege des Gehirns verlaufen. Denn für größere Proteine und Moleküle müsste es eine geeignete Route geben, um das ZNS verlassen zu können, da diese zum Passieren der Bluthirnschranke (BHS) zu groß sind. In dieser Arbeit konnte erstmals gezeigt werden, dass es auch eine Art lymphatischen Drainageweg aus der Hypophyse (HVL = Adenohypophyse) zu den tiefen und superfizialen, zervikalen Lymphknoten gibt. Daneben konnten auch die Abflussrouten des CSF und über die Nase bestätigt werden. Darüber hinaus wurde versucht zu ermitteln, wie schnell über die jeweiligen Drainagemöglichkeiten extrazelluläre Substanzen und Flüssigkeiten (Liquor) aus dem Gehirn gelangen und weiter in die peripheren Lymphknoten des Halses abtransportiert werden können. Hierbei sollte auch dargestellt werden, ob diese Mechanismen nur passiven Vorgängen zuzuordnen sind, oder ob es auch Hinweise auf aktive, zellulär gesteuerte Prozesse gibt. Wahrscheinlich spielen hier Makrophagen, die hirneigene Proteine und Antigene aufnehmen und in Lymphknoten transportieren und präsentieren, eine wesentliche Rolle. Obwohl es die BHS gibt, die als eine physiologische Barriere- und Filterfunktion zwischen dem Blutkreislauf und dem ZNS fungiert, existieren dennoch autoimmun bedingte Krankheitsbilder des ZNS wie die Autoimmun-Hypophysitis oder die MS. Es ist daher wahrscheinlich, dass es möglicherweise auch mehrere Mechanismen (passiv und aktiv) geben könnte, über die hirneigene Proteine und Substanzen aus dem Gehirn zu den Lymphknotenstationen gelangen können. Über das Lymphsystem wäre wiederum eine Aktivierung des Immunsystems naheliegend, sodass die aus dem ZNS entstammenden Proteine eine Immunantwort initiieren und so autoimmune Krankheitsprozesse einleiten könnten. Für das Verständnis von Erkrankungen wie der MS, der Autoimmunhypophysitis oder aber auch des Morbus Alzheimer wären genauere Kenntnisse über die funktionellen Hintergründe eines „hirneigenen lymphatischen Systems“ von großer klinischer und therapeutischer Relevanz. Auf der Grundlage der Ergebnisse dieser Arbeit könnte das Entstehen von Autoimmunerkrankungen des ZNS und eben auch der Hypophyse eine Erklärung finden, da auch Antigene dem Immunsystem über die beschriebenen Abflusswege zu den peripheren Lymphknotenstatioen des Halses gelangen und präsentiert werden können. Weiterhin besteht die Vermutung, dass zusätzlich auch Makrophagen aktiv durch ihre antigenpräsentierenden Eigenschaften an solchen Immunreaktionen beteiligt sind, jedoch sehr wahrscheinlich in verzögerter Weise im Vergleich zu den schnelleren passiven Drainagerouten des ZNS.

Thesis (PhD)-- Stellenbosch University, 2013. / ENGLISH ABSTRACT: Although the effect of inhaled corticosteroids (ICS) on the hypothalamic- pituitary-adrenal axis (HPA) has been regarded as a “benign physiological response”, a survey published in 2002 suggested that adrenal crisis is more common in asthmatic children on ICS than previously thought. Relying on clinical features to detect chronic adrenal insufficiency secondary to corticosteroids may not be wise, as these are non-specific and can therefore easily be missed. Accurate biochemical assessment of the whole axis to detect subclinical HPA suppression (HPAS) is thus desirable. A review of the literature indicates that basal adrenal function tests, including plasma cortisol profiles, do not identify which children can appropriately respond to stress. There is no evidence to suggest that the degree of the physiological adjustment of the HPA to ICS and/or nasal steroids (by reducing basal cortisol production), predicts HPAS. Cortisol profiles should therefore only be used to demonstrate differences in systemic activity of various ICS and delivery devices. Only two tests, considered as gold standard adrenal function tests [the insulin tolerance test (ITT) and the metyrapone test] can assess the integrity of the whole axis. / AFRIKAANSE OPSOMMING: Die outeurs van ´n opname wat in 2002 gepubliseer is stel voor dat ´n bynierkrisis meer algemeen by asmatiese kinders, wat inhalasie kortikosteroïede ontvang, voorkom as wat voorheen gedink is. Dit is strydig met die gevestigde opvatting dat die effek van IKS op die hipotalamiese-hipofise-bynier-as (HHB) ’n “goedaardige fisiologiese reaksie” is. Die kliniese kenmerke van kroniese bynierontoereikendheid sekondêr tot die gebruik van kortikosteroïede (KS) is nie-spesifiek en gevolglik onbetroubaar. ´n Akkurate biochemiese toets van subkliniese HBB onderdrukking (HHBO) sou gevolglik waardevol wees. ´n Literatuur oorsig toon dat toetse van basale bynierfunksie, insluitend plasma kortisol (K) profiele, nie kinders uitken wat toepaslik op stres sal reageer nie. Daar is geen bewyse dat die graad van fisiologiese aanpassing van die HHB, soos aangedui deur laer K-vlakke, na die gebruik van IKS en/of nasale steroïede (NS), HHBO voorspel nie. Serum K profiele is dus slegs van waarde om die sistemiese aktiwiteit van verskillende IKS en toedieningsstelsels te ondersoek. Slegs twee toetse, naamlik die insulien toleransie toets (ITT) en die metyrapone -(MTP)-toets (wat beide as die goue standaard van bynier funksie beskou word), kan die integriteit van die hele as meet. / Stellenbosch University / Medical Research Council / SA Thoracic Society / Harry Crossley Foundation / Red Cross Children’s Hospital.

L’embryologie de la surrénale humaine et la pathogénie de l’hypoplasie congénitale des surrénales sont mal connues et complexes. L’une pouvant expliquer l’autre, nous présentons, à partir de 119 fœtus humains normaux de12 à 36 semaines de développement (SD), une étude cinétique histologique et moléculaire de la surrénale fœtale et nous décrivons 3 cas d’hypoplasie surrénalienne. Après un rappel des mécanismes moléculaires connus, régulant ensemble l’embryologie surrénalienne, gonadique et hypophysaire et ceux de l’hypoplasie surrénale congénitale, nous présentons nos résultats. Dans la corticosurrénale humaine fœtale normale, les cellules du cortex permanent prolifèrent et, dès la 12ème SD, expriment la NCAM, la 3β-HSD et la P450 c21. Elles ont la capacité de synthétiser des minéralocorticoïdes et/ou du cortisol. Les cellules du cortex fœtal ne prolifèrent pas et expriment ni la 3β-HSD, ni la NCAM. La médullosurrénale est formée par des neuroblastes immatures (CgA-, NCAM+) qui migrent et prolifèrent de la périphérie vers le centre de la glande, où ils se différencient en neuroblastes matures (CgA+). Dans les deux cas d’hypoplasie surrénale de type anencéphalique, avec absence de mutation de DAX-1 et de SF-1, la dysembryoplasie surrénalienne est probablement d’origine hypophysaire, par absence de cellules gonadotropes. Dans le 3ème cas, jamais décrit, associant un RCIU, une hypoplasie surrénalienne congénitale, une ambiguïté sexuelle, une absence de différenciation des cellules antéhypophysaires, l’absence de mutation de gènes connus (DAX-1, SF-1, SRY, FGF9, SOX2, SOX3, SOX5 et SOX9) suggère l’existence de nouveaux gènes régulant la différenciation précoce de ces trois glandes endocrines. / The embryology of the human adrenal and the pathogenesis of the hypoplasia adrenal congenital are complex and not well known. We studied the histological and functional characteristics of the fetal human adrenal in normal fetuses aged 12 to 36 weeks development (WD) and presented three cases of congenital adrenal hypoplasia with pituitary abnormalities. In the human fetal adrenal cortex, only the cells of the definitive zone proliferate from the 12th WD. We observe the expression of an adherence protein (NCAM) and of two steroidogenesis enzymes (3β-HSD and P450 c21) in the definitive zone cells, attesting to the capacity of these cells to synthesize mineralocorticoids and/or cortisol. In the fetal zone, only P450 c21 immunoreactivity is detected. The adrenal medulla is formed by immature neuroblasts (CgA -, NCAM+) which migrate and proliferate from the periphery toward the center of the gland, where they differentiate in mature neuroblasts (CgA+). In the two cases of congenital adrenal hypoplasia of anencephalic type without DAX 1 and SF-1 mutation, the cause is probably from pituitary origin and related to absence of gonadotrope cells. In the third case of congenital adrenal hypoplasia, never described, combining a retard intrauterine development, a congenital adrenal hypoplasia, sexual ambiguity, pituitary abnormalities. The absence of mutation of known genes (DAX -1, SF -1, SRY, FGF9, SOX2, SOX3, SOX5 and SOX9) suggests the existence of new genes regulating early differentiation of the adrenal, the gonad and the pituitary.

Os piretróides sintéticos, em especial os do tipo II, como a cialotrina, são extensivamente utilizados para o controle de uma ampla variedade de ectoparasitas que acometem os animais de produção. Entretanto, no Brasil e em outros países, sua utilização vai além da saúde animal, sendo utilizados também em saúde pública, no controle de diversos vetores, como é o caso do vetor da dengue, dentre outros. Visto que a cialotrina modifica a atividade de macrófagos peritoneais, o objetivo deste trabalho foi investigar os prováveis mecanismos através dos quais este piretróide modifica a atividade destas células. Os presentes resultados, analisados em seu conjunto, mostram de maneira inequívoca que a cialotrina tem um efeito direto e/ou indireto sobre a atividade de macrófagos peritoneais. Especificamente, observou-se neste trabalho que o praguicida causou em ratos: 1 ? marcação fos positiva em neurônios do núcleo paraventricular do hipotálamo (NPH), após a dose de 3,0 mg/kg/dia; 2 - diminuição do percentual e intensidade de fagocitose de macrófagos peritoneais ativados e avaliados por citometria de fluxo; 3 - diminuição dose-dependente da produção de nitrito (NO2); 4 ? diminuição do percentual e intensidade de fagocitose de macrófagos peritoneais ativados, em ratos adrenalectomizados e/ou tratados com metirapona (inibidor da síntese de corticosterona) e RU 486 (antagonista de receptores glicocorticóides) com a finalidade de modular os níveis de glicocorticóides, e tratados com 3,0 mg/kg/dia de cialotrina; 5 ? aumento dos níveis de noradrenalina hipotalâmica em animais tratados com a dose de 3,0mg/kg/dia de cialotrina; 6 - diminuição do percentual e intensidade de fagocitose, bem como diminuição da produção de nitrito de macrófagos peritoneais ativados, em ratos simpatectomizados químicamente com 6-OHDA; 7 - diminuição dose dependente do percentual e intensidade de fagocitose, bem como da produção de nitrito de macrófagos peritoneais ativados e tratados in vitro com 10 e 100 nM de cialotrina. No entanto, não observamos: 1 ? alterações na produção de nitrito realizada por macrófagos peritoneais ativados, em ratos adrenalectomizados e/ou tratados com metirapona e RU 486; 2 - alterações na viabilidade celular induzida pelo tratamento in vitro com a cialotrina na concentração de 10 e 100 nM e 3 ? alterações nos efeitos da cialotrina sobre a atividade de macrófagos tratados in vitro com os ligantes de receptores benzodiazepínicos periféricos. Em conjunto, os presentes dados mostram que a cialotrina interfere com a atividade de macrófagos por atuar indiretamente, através da ativação do eixo Hipotálamo-Hipófise-Adrenal (HHA), e/ou diretamente sobre os mesmos modulando sua atividade. É muito provável que o efeito resultante do tratamento in vivo com este praguicida esteja ligado à somatória destas ações / Synthetic pyrethroids, particularly those of type II, such as cyhalothrin, are extensively used in agriculture for the control of a broad range of ectoparasites in farm animals. However, in Brazil and some other countries, these pyrethroids have also been used in public health, for the control of insects that are known to be vectors of diseases such as dengue. Since it has been suggested that cyhalothrin alters activity of peritoneal macrophages, the objective of our study was to investigate the putative mechanisms for the changes induced by pyrethroid in these cells. The results presented here show, in an unequivocal manner, that cyhalothrin has a direct or indirect (or both) effect on the activity of peritoneal macrophages. We specifically observed in this work that this pesticide induced in rats: 1- Fos-positive immunostaining in neurons of the paraventricular nucleus of the hypothalamus (NPH), after 3.0 mg/kg/day; 2 ? a reduction in the percentage and intensity of phagocytosis by activated peritoneal macrophages, evaluated by flow cytometry; 3 ? a dose-dependent reduction in nitrite production (NO2); 4 ? a reduction in the percentage and intensity of phagocytosis by activated peritoneal macrophage from adrenalectomized rats treated or not with metirapone (inhibitor of corticosterone synthesis) or RU 486 (antagonist of glicocorticoids receptors) with the propose of modulating the levels of glicocorticoids, and treated with 3.0 mg/kg/day of cyhalothrin; 5 ? an increase in the hypothalamic levels of noradrenaline in rats treated with 3.0 mg/kg/day of cyhalothrin; 6 ? a reduction in the percentage and intensity of phagocytosis and also a decrease in the production of nitrite by activated peritoneal macrophages, after chemical sympatectomy with 6-OHDA; 7 ? a dose-dependent reduction of the percentage and intensity of phagocytosis, and also a decrement in nitrite production by activated peritoneal macrophages treated in vitro with 10 and 100 nM of cyhalothrin. However, we found no differences on: 1 ? nitrite production by activated peritoneal macrophages after adrenalectomy, treated or not with metirapone or RU 486; 2 ?cell viability of peritoneal macrophages treated in vitro with 10 and 100 nM of cyhalothrin, and 3 ? the effects of cyhalothrin on macrophage activity after in vitro treatment with peripheral benzodiazepine receptor ligands. Altogether, the present results show that cyhalothrin interferes with the activity of peritoneal macrophages by acting indirectly, via activation of the hypothalamus-pituitary-adrenal axis, or directly on these cells, altering their activity. As a matter of fact, it is quite possible that the results of in vivo cyhalothrin treatment on macrophage activity would be related to the combined effect of these direct and indirect influences

Neste estudo foi verificado que a hipófise de S. brasiliensis era composta por dois tecidos, a neuro hipófise (NH) e a adeno-hipófise (AH). Nesta última, foram distinguidos sete tipos celulares. Na RPD foram detectadas as células adrenocorticotrópicas e prolactínicas, na PPD as células gonadotrópicas, somatotrópicas e tireotrópicas, e na PI, as células melanotrópicas e somatolactínicas. Foi evidenciada única célula gonadotrópica, produtora de LH e FSH. O primórdio da hipófise foi detectado 12 horas após a eclosão (hpe), a NH com 72 hpe e o início da formação do pedúnculo com 300 hpe. Nos juvenis (600 hpe), a hipófise apresentou uma morfologia semelhante àquela observada nos adultos. As células prolactínicas foram detectadas com 12 hpe, juntamente com as células adrenocorticotrópicas e melanotrópicas, seguidas das somatotrópicas e somatolactínicas, com 36 hpe. Por outro lado, nas larvas e juvenis foram detectadas duas populações distintas de células gonadotrópicas, as células produtoras de FSH foram detectadas com 600 hpe, enquanto as produtoras de LH com 120 hpe. / This study showed that the pituitary gland of S. brasiliensis was formed by two tissues, neurohypophysis and adenohypophysis. In the latter, seven cell types were distinguished. In RPD, prolactin and adrenocorticotropic cells were present. In PPD, gonadotropic, somatotrope and thyrotropic cells were detected, and in PI, somatolactin and melanotropic cells were found. Interestingly, was detected a single gonadotropic cell responsible for producing both gonadotropins. The primordium of the pituitary gland was detected 12 hours after hatching (hah), the neurohypophysis was detected 72 hah and formation of the stalk 300 hah. In juveniles, 600 hah, the pituitary showed a similar morphology to that observed in adults of this species. Prolactin cells were detected 12 hah together with adrenocorticotropic and melanotropic cells, followed by somatotropic and somatolactin cells 36 hah. Unlike adults, larvae and juveniles have shown two distinct populations of gonadotropic cells. FSH-producing cells were detected 600 hah, while LH-producing cells were detected 120 hah.

INTRODUÇÃO: A prolactina é um hormônio polipeptídico, que possui reconhecida ação sistêmica, principalmente na fisiologia da reprodução, porém, seu desequilíbrio, em especial a hiperprolactinemia, é cada vez mais frequente na prática clínica. Apesar de ser um distúrbio relativamente comum, ainda existem dúvidas quanto aos efeitos moleculares da hiperprolactinemia no trato genital, particularmente no útero, e também na hipófise. O presente estudo teve por objetivo verificar os efeitos da hiperprolactinemia induzida pela metoclopramida na expressão gênica da prolactina e de seus receptores no útero e na hipófise de camundongo fêmea. MÉTODOS: Utilizaram-se 49 camundongos fêmeas (Wistar), randomicamente divididas em 7 grupos contendo 7 animais cada: 1) SS não ovariectomizadas que receberam solução salina (veículo); 2) M não ovariectomizadas tratadas com metoclopramida; 3) OSS ovariectomizadas tratadas com solução salina (veículo); 4) OM ovariectomizadas tratadas com metoclopramida; 5) OME ovariectomizadas tratadas com metoclopramida e 17-estradiol; 6) OMP ovariectomizadas tratadas com metoclopramida e progesterona micronizada; 7) OMEP ovariectomizadas tratadas com metoclopramida, 17-estradiol e progesterona micronizada. Após 50 dias os animais foram sacrificados sendo retirados o útero e a hipófise de cada animal para extração do ácido ribonucleico total, que foi utilizado para a síntese de ácido desoxirribonucleico complementar e avaliação da expressão gênica da prolactina e das diferentes isoformas de seus receptores, por reação em cadeia da polimerase em tempo real. RESULTADOS: Na hipófise, em animais não ovariectomizados, o tratamento com metoclopramida aumentou a expressão do gene que codifica a prolactina em relação ao tratamento apenas com o veículo. Nos animais castrados, a progesterona isoladamente ou associada ao estrogênio determinou o incremento do RNA mensageiro da prolactina em relação aos outros animais castrados que receberam outras combinações de tratamento. Este efeito foi semelhante ao da metoclopramida em animais com os ovários intactos. Em relação ao receptor de prolactina, o estrogênio e a progesterona, isoladamente, foram responsáveis pelo incremento da isoforma S2. No útero houve aumento na expressão de RNA mensageiro de prolactina após tratamento com metoclopramida ou com tratamento isolado ou combinado de estrogênio e progesterona. A ovariectomia determinou a redução da expressão das isoformas S1 e S2 do receptor de prolactina de todas as isoformas estudadas. Já o tratamento estroprogestativo determinou elevação da formas S3 e L do receptor, enquanto com a progesterona isoladamente causou apenas o incremento da forma L do receptor da prolactina no útero dos animais castrados. CONCLUSÕES: Nossos dados sugerem que o tratamento com metoclopramida altera de forma diferente a expressão de prolactina e de seus receptores quando se compara o resultado da hipófise em relação ao útero em camundongos fêmeas castrados e tratados com esteróides sexuais / INTRODUCTION: Prolactin is a polypeptide hormone with a recognized systemic action mainly on reproductive physiology. However, prolactin imbalance, particularly hyperprolactinemia, is increasingly more frequent in clinical practice. Although it is a comparatively common disorder, there are still doubts about the molecular effects of hyperprolactinemia on the genital tract especially in the uterus and the pituitary. The present study aimed at verifying the effects of metoclopramide-induced hyperprolactinemia on the gene expression of prolactin and its receptors in the uterus and pituitary of the female mouse. METHODS: Forty-nine female Wistar mice were randomized to 7 equal-sized groups as follows: 1) SS nonoophorectomized mice treated with saline solution (vehicle); 2) M nonoophorectomized mice treated with metoclopramide; 3) OSS oophorectomized mice treated with saline solution (vehicle); 4) OM oophorectomized mice treated with metoclopramide; 5) OME oophorectomized mice treated with metoclopramide and 17-estradiol; 6) OMP oophorectomized mice treated with metoclopramide and micronized progesterone; 7) OMEP oophorectomized mice treated with metoclopramide, 17-estradiol, and micronized progesterone. The animals were sacrificed 50 days after the end of the treatment, and the uterus and pituitary of each animal were removed for extraction of total ribonucleic acid, which was then used for synthesizing complementary deoxyribonucleic acid and for evaluating the gene expression of prolactin and the different isoforms of its receptors by the real-time polymerase chain reaction. RESULTS: In the pituitary of the nonoophorectomized mice, the treatment with metoclopramide against that with vehicle alone increased the expression of the prolactin-encoding gene. In the castrated animals, progesterone by itself or in conjunction with estrogen determined a raise in prolactin messenger RNA as opposed to the two other treatments with different combinations. This effect was similar to that produced by metoclopramide in animals with intact ovaries. Estrogen and progesterone, acting independently of each other, were responsible for the increase in the S2 isoform of the prolactin receptor. In the uterus, there was heightened expression of prolactin messenger RNA under the effect of the treatment with metoclopramide or with estrogen and/or progesterone. Oophorectomy caused a greater reduction in expression of the prolactin receptor S1 and S2 isoforms than in the other isoforms. However, the combined estrogen plus progesterone treatment led to an increase in the S3 and L forms of the receptor, while progesterone alone resulted solely in a higher expression of the L form of the prolactin receptor in the endometrium of the castrated mice. CONCLUSION: Our data suggest that metoclopramide treatment induces different changes in the expression of prolactin and its receptors according to whether the effect occurs in the pituitary or the uterus of castrated female mice treated with sex steroids

Este estudo analisa a expressão dos subtipos de receptores da somatostatina (SSTR) em 39 adenomas secretores de GH. Em 19 pacientes acromegálicos, a resposta clínica, laboratorial e radiológica ao análogo da somatostatina (AS) octreotide-LAR foi comparada à expressão dos SSTR. O SSTR mais freqüentemente expresso foi o SSTR5, seguido pelos SSTR3, SSTR2, SSTR1 e SSTR4. O SSTR1 e SSTR2 foram mais expressos nos pacientes que normalizaram GH e IGF-I. Houve correlação positiva entre o grau de redução tumoral e a expressão dos SSTR1, SSTR2 e SSTR3. Portanto, AS específicos para os SSTR´s poderão contribuir para o tratamento de acromegálicos resistentes aos AS atualmente disponíveis / This study aimed to analyze the expression of somatostatin receptor subtypes (SSTR) in 39 GH-secreting pituitary adenomas. In 19 acromegalics the clinical, laboratorial and radiological responses to the somatostatin analog (SA) octreotide-LAR were compared to SSTR´s expression. The most expressed SSTR was SSTR5, followed by SSTR3, SSTR2, SSTR1 and SSTR4. SSTR1 and SSTR2 were more expressed in patients who achieved GH and IGF-I normalization. There was a positive correlation between the degree of tumor reduction with SSTR1, SSTR2 and SSTR3 expression. Therefore, the development of specific SA could contribute to treatment improvement in resistant acromegalics patients to available SA