Rozprawy doktorskie na temat „Endocrine glands – Diseases”

'Endocrine disrupting chemicals (EDCs) are compounds found in the environment that have the potential to disrupt normal endocrine function. Estrogenic EDCs (e-EDCs) is a subclass of EDCs and is defined as substances contaminating the environment that may mimic or inhibit the effect of endogenous estrogen and therefore may influence developmental and reproductive health in humans and animals. The aim of this study was to develop, validate and implement a battery of in vitro and in vivo screening assays for e-EDCs. The study was concluded by implementing this battery of assays to assess the Eerste River, South Africa at three sampling sites, namely Jonkershoek, Stellenbosch sewage treatment works (STW) effluent and Spier for e-EDCs. The control site, Jonkershoek contained very low levels of estrone. Water from this site showed no estrogenic activity when the E-screen and the ER_ induction in MCF-7 cells. Some of the water samples collected at this site tested positive for estrogenicity when analysed with the juvenile tilapia VTG assay, whereas the rest were negative. The estrone levels in the sewage effluent extracts as well as Spier were significantly higher. The assay using ER_ protein induction by the MCF-7 cell line, the MCF-7 proliferation assay and the tilapia in vivo screen for estrogenicity showed that these samples are estrogenic. Results obtained for estrogenicity at the three different sampling sites for each of the assays in the battery were comparable. In this study we developed, validated and also implemented a battery of assays encompassing both in vitro and in vivo assays, based on different biological mechanisms, to detect estrogenic EDCs. To our knowledge, this is the first study that has used a battery of bioassays to specifically assess a South Africa river for estrogenicity...'

Congenital hypothyroidism (CH) is the most frequent of the rare endocrine diseases (e.g. Addison's disease, Cushing's syndrome, Congenital adrenal hyperplasia.), which affects 1:2000 – 4000 newborns. If not immediately diagnosed after birth, thyroid hormones deficiency causes severe defects in brain and skeletal development leading to a complex clinical scenario called cretinism. CH can be due to a defective synthesis of thyroid hormones (dyshormonogenesis) or an abnormal embryonic development of the gland. Data obtained using knockout mouse models have shown the pivotal role of four specific transcription factors (NKX2.1, PAX8, FOXE1 and HHEX) for the correct organogenesis or function of the gland. Although mutations in those genes have been identified in few cases of CH patients, the pathogenetic mechanisms remain still elusive in the vast majority of CH cases (95%).

For the identification of new genes and molecular events controlling thyroid organogenesis it would be useful to develop an in vitro cellular model to recapitulate thyroid embryogenesis in a dish. Embryonic Stem Cells (ESCs) have recently emerged as system model to recapitulate the embryogenesis of several tissues in vitro.

Induced overexpression of defined transcription factors has been shown to have a directing effect on the differentiation of pluripotent stem cells into specific cell types. In this thesis I show that a transient overexpression of the transcription factors NKX2.1 and PAX8 is sufficient to direct the differentiation of murine ESCs into thyroid follicular cells (TFC) and promotes in vitro self- assembly of TFC into three-dimensional follicular structures, when associated to a subsequent thyrotropin (TSH) treatment. Cells differentiated by this protocol showed significant iodide organification activity, a hallmark of thyroid tissue function. Importantly, athyroid mice grafted with mESC-derived thyroid follicles show normalization of plasma T4 levels with concomitant decrease of plasma TSH. In addition, a full normalization of body temperature at 4 weeks after transplantation was observed. Together, these data clearly demonstrate that grafting of our mESC-derived thyroid cells rescues the hypothyroid state and triggers symptomatic recovery along with the normalization of plasma hormone concentrations. The high efficiency of TFC differentiation and follicle morphogenesis in our system will provide an unprecedented opportunity for future studies to decipher regulatory mechanisms involved in embryonic thyroid development, a major research need towards an improved understanding of the molecular mechanisms underlying congenital hypothyroidism, the most common congenital endocrine disorder in humans.
Doctorat en Sciences biomédicales et pharmaceutiques
info:eu-repo/semantics/nonPublished

Dissertation (PhD)--University of Stellenbosch, 2003.
ENGLISH ABSTRACT: Concern has increased that widespread adverse effects are occurring in humans and wildlife populations as a result of exposure to environmental chemicals (mostly man-made) that disrupt the normal functioning of the endocrine system. Many pharmaceutical, agricultural and industrial chemicals, acting as endocrine modulating compounds (EDCs), have been detected in an accumulated form in food, in drinking water and in the environment. Although the levels of these chemicals can be measured analytically, it is important to evaluate biological activity. For this, animal models are used and relevant bioassays developed. These assays are based on biological markers, which are specific xenobiotically-induced physiological responses and are usually deviations in cellular or biochemical components, processes or structures. Vitellogenin is a large protein complex, produced in the liver under estrogen control and circulates in the plasma, destined for incorporation into the developing oocyte in female oviparous vertebrates. Since vitellogenin production is low or nonexistent in males, its presence may therefore be interpreted as evidence of exposure to estrogenic environmental contaminants. In this study the African Clawed Frog, Xenopus laevis was used as model to study the effects of estrogens on biochemical and physiological parameters associated with vitellogenesis. As a starting point the seasonal female reproductive cycle in a natural Xenopus laevis population in terms of ovarian state, plasma vitellogenin and plasma estrogen levels was studied. It was shown that plasma vitellogenin and estrogen levels were seasonal and correlated to a seasonal ovarian cycle, which peaked during spring. However, although seasonality existed, there were reproductively active individuals at any time during the year. Increases in plasma estrogen levels predated increases in plasma vitellogenin levels and ovarian maturation. Lipoprotein profiles, as well as plasma cholesterol, triglyceride and phospholipid concentrations were determined and it was shown that estrogen affected these in such a way that these parameters could be used as additional markers in estrogen contamination studies. In order to develop an in vitro bioassay to screen for estrogenic activity, the use of hepatic tissue cultures was investigated. Optimal culture conditions were established and increased sensitivity in the estrogenic response was obtained by using liver slices from male frogs that were pre-treated with estrogen. Validation studies proved that this bioassay could be employed for routine screening of water and chemical samples. In order to refine the Xenopus laevis vitellogenin ELISA and liver slice bioassay, existing polyclonal anti-vitellogenin antibodies were replaced by culturing monoclonal antibodies. Selected antibodies were characterised and ELISAs developed and validated. This study showed that the newly developed Xenopus laevis vitellogenin ELISA and liver slice bioassay have the potential to be employed in environmental monitoring programmes.
AFRIKAANSE OPSOMMING: Daar is toenemende besorgdheid dat afwykings in mens- en dierbevolkings voorkom as gevolg van blootstelling aan chemikalieë (hoofsaaklik mensgemaak) in die omgewing wat die normale werking van die endokrienstelsel versteur. Verskeie farmaseutiese, landbou- en industriële chemikalieë, wat as endokrienversteurders optree, is in die omgewing gevind. AI kan die vlakke van hierdie stowwe analities bepaal word, is dit belangrik om hulle biologiese aktiwiteit te evalueer. Hiervoor word diermodelle aangewend om toepaslike toetse daarvoor te ontwikkel. Hierdie toetse word baseer op biologiese merkers, spesifieke xenobioties-geïnduseerde fisiologiese reaksies, en is gewoonlik afwykings van sellulêre- of biochemiese komponente, -prosesse of - strukture. Vitellogeen ('n dooiervoorloper) is 'n lipoproteïenkompleks wat, onder estrogeenbeheer, in die lewer vervaardig word en in die plasma sirkuleer vir uiteindelike inkorporasie in ontwikkelende oësiete van vroulike, ovipare werweldiere. Aangesien daar min of geen vitellogeen in manlike diere geproduseer word, is die teenwoordigheid daarvan 'n aanduiding dat die dier aan estrogeniese omgewingsbesoedeling blootgestel is. In hierdie studie is die Platanna, Xenopus laevis, as model gebruik om die gevolge van estrogene op biochemiese en fisiologiese veranderlikes, wat met vitellogenese geassosieer word, te bestudeer. As vertrekpunt is die seisoenale voortplantingsiklus van die wyfie, in terme van vitellogeen en -estrogeen vlakke in die plasma en aktiwiteit van die ovaria bepaal. Daar is aangetoon dat die estrogeen- en vitellogeenvlakke in die plasma met die ovariumsiklus, wat gedurende die lente hoogtepunte bereik, korreleer. Alhoewel daar seisoenaliteit bestaan, was daar dwarsdeur die jaar wyfies wat ovarium dooierneerlegging getoon het. Verhoging in estrogeenvlakke het vitellogeenpieke en rypwording van die ovaria voorafgegaan. Lipoproteïenprofiele, sowel as die cholesterol- , trigliseried- en fosfolipiedkonsentrasies in die plasma is bepaal en daar is aangetoon dat estrogeen hierdie medeveranderlikes in só 'n mate affekteer dat hulle as bykomende biomerkers vir estrogeenblootstelling in besoedelingstudies gebruik kan word. In vitro Xenopus laevis lewersnitte in die weefselkultuur omgewing is ondersoek om 'n biotoets te onwikkel vir die gebruik in vinnige estrogenisiteits-toetsing van watermonsters en chemise stowwe. Die mees gunstige kultuurtoestande is bepaal en die sensitiwiteit van estrogeenreaksies is verhoog deur lewer van mannetjies, wat vooraf met estrogeen behandel is, te gebruik. Hierdie biotoets se geldigheid is gestaaf en kan in roetine eerstevlaktoetsing van watermonsters gebruik word. Die gebruik van poliklonale teenliggaampies in 'n bestaande enzyme-linked immunosorbent assay (ELISA) is vervang deur spesiaal-ontwikkelde monoklonale anti-Xenopus laevis vitellogeen teenliggaampies. Uitgesoekte teenliggaampies, spesifiek teen die estrogeengeïnduseerde proteïene, is gekarakteriseer en ELISAs saamgestel en die geldigheid gestaaf. Hierdie studie het aangetoon dat die nuut-onwikkelde Xenopus laevis vitellogeen-ELISA en lewerkultuurbiotoets die potensiaal het om In omgewingsmoniteringprogramme gebruik te word.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
This thesis involved the study of the adrenal and thyroid glands. The study of adrenals was based, primarily, on epidemiological and pathological aspects of the lesions observed in the glands of 300 dogs necropsied in the Laboratório de Patologia Veterinária at the Universidade Federal de Santa Maria (LPV/UFSM). For this purpose, a prospective study was performed from March 2012 to June 2013, totaling 16 months. Of the 300 dogs examined, 82% had one or more adrenal lesions. Only 18% of these dogs did not have adrenal changes. The hyperplastic lesions were the most prevalent, comprising 91.5% of the cases, followed by degenerative (26%), circulatory (20.3%), neoplastic (15%), inflammatory (4.1%) and other lesions (14.2%). Most of the animals with hyperplastic lesions consisted of females, elderly and purebred dogs. Fatty degeneration was the most common degenerative lesion (54.7%). Among the neoplastic changes, primary tumors were the most prevalent (75.7%), with a prevalence of adenomas (57.1%), followed by pheochromocytomas (25%) and cortical carcinomas (17.9%). Metastatic tumors corresponded to 24.3% of all cases of neoplastic changes. The adrenal glands were the site of metastasis of mammary gland carcinoma, osteosarcoma, melanoma, lymphoma, cholangiocarcinoma, histiocytic sarcoma and lung adenocarcinoma. Under "other lesions", adrenal of stress was the most prevalent change (37.1%). It was characterized histologically by a cortex with irregular appearance. For the study of thyroid lesions, it was reviewed the protocols of biopsies and necropsies performed at the LVP/UFSM between 2004 and 2014, and all cases of thyroid tumors were selected. Epidemiological data, clinical significance of tumors, presence of cervical swelling, affected lobes, tumor size, presence and location of metastasis and histological pattern of the neoplasm. It were recorded 26 tumors of the thyroid gland. Of the affected dogs, 53.9% were females and 46.1% were males; 65.4% were purebred. Of these, Boxers were the most affected (15.4%). In 23 protocols the age was informed; 60.9% of the dogs were elderly and 39.1% were adults. The protocols evaluation allowed establishing that 38.5% of the animals had shown cervical swelling, at the time of clinical examination. From the dogs that underwent to necropsy, only 13.6% died or were euthanized due to complications caused by metastases of thyroid tumors. Thus, in 86.4% of the dogs, the neoplasms were considered as incidental necropsy findings. Glandular unilateral involvement occurred in 75% of the dogs. The histological diagnosis of primary tumors showed that 61.5% of the neoplasms were classified as follicular cell carcinomas, 23.1% as follicular cell adenomas, 7.7% as carcinosarcomas and 7.7% as tumors of C cell. Additionally, two cases of thyroid follicular cell carcinoma in dogs that developed myelopathy secondary to cervical spinal epidural metastasis were minutely evaluated, and separately reported.
Esta tese envolveu o estudo das adrenais e da tireoide. O estudo das adrenais baseou-se, principalmente, nos aspectos epidemiológicos e patológicos das lesões observadas nas glândulas de 300 cães necropsiados no Laboratório de Patologia Veterinária da Universidade Federal de Santa Maria (LPV/UFSM). Esse estudo foi realizado de forma prospectiva, de março de 2012 a junho de 2013. Dos 300 cães analisados, 82% tinham uma ou mais alterações nas adrenais e 18% não tinham alterações. As lesões hiperplásicas foram as mais prevalentes, com 91,5% das ocorrências, seguidas de lesões degenerativas (26%), circulatórias (20,3%), neoplásicas (15%), inflamatórias (4,1%) e outras lesões (14,2%). A maioria dos animais que apresentou lesões hiperplásicas era constituída de fêmeas, idosos e cães com raça definida. A degeneração gordurosa foi a lesão degenerativa mais frequente (54.7%). Entre as alterações neoplásicas, tumores primários foram mais prevalentes (75,7%), com predomínio de adenomas (57,1%), seguido de feocromocitomas (25%) e carcinomas corticais (17,9%). Os tumores metastáticos perfizeram 24,3% dos casos de alterações neoplásicas. As adrenais foram sítio de metástase de carcinoma mamário, osteossarcoma, melanoma, linfoma, colangiocarcinoma, sarcoma histiocítico e adenocarcinoma pulmonar. No item outras lesões , adrenal de estresse foi a alteração mais prevalente (37,1%). Essa alteração era caracterizada histologicamente por córtex de aspecto variegado. Para o estudo das lesões de tireoide foram revisados os protocolos de biópsias e necropsias do LPV/UFSM entre os anos de 2004 e 2014 e todos os casos de tumores tireoidianos foram selecionados. A partir disso, foram anotados dados epidemiológicos, significado clínico dos tumores, presença de aumento de volume cervical, lobos afetados, tamanho, presença e localização de metástases e padrão histológico dos neoplasmas. Foram diagnosticados 26 neoplasmas tireoidianos. Dos cães acometidos, 53,9% eram fêmeas e 46,1% eram machos; 65,4% eram cães com raça definida. Desses, Boxers foram os mais afetados (15,4%). Dos 23 protocolos em que a idade foi informada, 39,1% eram cães adultos e 60,9% eram idosos. A avaliação dos protocolos permitiu constatar que 38,5% dos animais haviam apresentado aumento de volume cervical no momento do exame clínico. Apenas 13,6% dos cães necropsiados morreram ou foram submetidos à eutanásia em decorrência de complicações causadas por metástases dos tumores de tireoide. Dessa forma, em 86,4% dos cães os neoplasmas foram considerados achados incidentais de necropsia. O acometimento unilateral da glândula ocorreu em 75% dos cães. Os diagnósticos histológicos dos tumores primários permitiram constatar que 61,5% dos neoplasmas eram carcinomas de células foliculares, 23,1% eram adenomas de células foliculares, 7,7% eram carcinossarcomas e 7,7% eram tumores de células C. Além disso, foram estudados de forma detalhada e relatados separadamente dois casos de carcinoma de células foliculares da tireoide em que os cães desenvolveram mielopatia secundária à metástase epidural cervical espinhal.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Tese (Doutorado em Tecnologia Nuclear)
IPEN/T
Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP
FAPESP:11/21708-6

[Truncated abstract] G-protein coupled receptors (GPCRs) form one of the largest superfamilies of cell-surface receptors and respond to a vast range of stimuli including light, hormones and neurotransmitters. Although structurally similar, GPCRs are regulated by many diverse proteins, which allow the specific functions of each receptor to be carried out. This thesis focussed on two well-documented GPCRs, the thyrotropin releasing hormone receptor (TRHR) and gonadotrophin-releasing hormone receptor (GnRHR), which control the thyroid and reproductive endocrine pathways respectively. Although each of these anterior pituitary receptors is responsible for distinct physiological responses, both are integral to normal development and homeostasis. This thesis focused on three areas of GPCR regulation: ?-arrestin recruitment, transcription factor regulation and receptor up-regulation. The role of the cytoplasmic protein, ?-arrestin, has perhaps been previously underestimated in GPCR regulation, but it is now increasingly apparent that ?-arrestins not only inhibit further G-protein activation and assist in GPCR internalisation but also act as complex scaffolding platforms to mediate and amplify downstream signalling networks for hours after initial GPCR activation. It is therefore becoming increasingly important to be able to monitor such complexes in live cells over longer time-frames. ... Members of the E2F transcription family have been previously identified by this laboratory as potential GnRHR interacting proteins, via a yeast-2-hybrid screen and BRET. This thesis further investigated the role of E2F family members and demonstrates that a range of GPCRs are able to activate E2F transcriptional activity when stimulated by agonist. However, despite GnRHR displaying robust E2F transcriptional activation upon agonist stimulation, this did not result in any conclusive evidence for functional regulation, although it is possible E2F may modulate and assist in GnRHR trafficking. Furthermore it is apparent that E2F family members are highly redundant, as small effects in GnRHR binding and cell growth were only observed when protein levels of both E2F4 and E2F5 were altered. During the course of the investigation into the effect of E2F transcription on GPCR function, it was evident that long-term agonist stimulation of GnRHR had a profound effect on its expression. As this was explored further, it became clear that this agonist-induced up-regulation was both dose- and time-dependent. Furthermore, altering levels of intracellular calcium and receptor recycling/synthesis could modulate GnRHR up-regulation. In addition, an extremely sensitive CCD camera has been used for the first time to visualise the luciferase activity attributed to GnRHR up-regulation. Overall, this thesis demonstrates the complex nature of GPCR regulation. For the first time, long-term BRET analysis on ?-arrestin interactions with both classes of GPCRs has been examined in a variety of cellular formats. This has given valuable insights into the roles of phosphorylation and internalisation on ?-arrestin interaction. Additionally, this thesis has revealed that prolonged agonist exposure increases receptor expression levels, which has major implications for drug therapy regimes in the treatment of endocrine-related disorders and tumours.

Thesis (MSc)--University of Stellenbosch, 2005.
ENGLISH ABSTRACT: Water quality analysis forms the basis in assessing and monitoring catchments. As urban development continuously increase, pollution sources increase in either point source (wastewater treatment works, industrial effluents) and/or non-point source origin (storm water discharge, domestic pollutants), accumulating pollutants in the environment. It was only recently discovered that certain pollutants have subtle disrupting effects on the endocrine system resulting in health related problems associated with the reproductive system and thyroid system (growth and development) of animals and potentially humans. Natural water resource management proves to include limited biological assays measuring endpoints for cytoxicity, inflammatory activity and endocrine disruption. The broad objective of this study was therefore to include several bioassays, not normally used in municipal (City of Cape Town) monitoring programmes, along with water quality data collected by the City of Cape Town. The Eerste/Kuils River catchment system, Western Cape, under the auspices of the City of Cape Town was chosen, and although this catchment does not contribute to drinking water resources, is subjected to a range of anthropogenic influences (industrial effluents, household wastewater, agricultural runoff). Within the short time-frame available for this study (six months) two months, July (following a dry summer and autumn season) and October (following a wet winter and early spring season) were selected for water quality monitoring. Spatial variation (with relevance to specific point and non-point contamination) among sampling sites were also obtained by choosing several (n=10) along the catchment. Specific aim of the study therefore included: Firstly (Chapter 2), the use of in vitro bioassays, lactate-dehydrolises assay (LDH) for cytotoxic activity, pro-inflammatory hormone Interleukin-6 (IL-6) secretion by human blood cells and a specific Salmonella ELISA for faecal contamination, in conjunction with routine chemical and biological (mostly microbiological) monitoring activities. The study indicated significant variation among sites in all microbiological measures as well in IL-6 secretion and Salmonella presence. Between months, variations were also evident in certain variables. Secondly (Chapter 3), two bioassays using the yolk precursor protein, vitellogenin (Vtg) as endpoint was implemented in a) an in vitro Xenopus laevis liver slice assay (five day exposure) and b) an in vivo Zebrafish (Danio rerio) bioassay (seven day exposure) assessing estrogenic activity in the Eerste/Kuils River catchment. Although estrogen spiked positive control water samples stimulated Vtg production in vitro as well as in vivo, no dramatic estrogenic activity was measured at any of the selected sites. Thirdly (Chapter 4), a bioassay using the thyroid controlled metamorphosis in Xenopus laevis tadpoles to assess effects on the thyroid hormonal system was implemented. Thyroid stimulatory activity, compared with a negative control sample, was measured at two sites along the catchment. Although the practical implementation of the tadpole semi-static exposure protocol (water replacement) proved to be labour intensive, all the added bioassays proved to be valuable tools to add valuable information regarding water quality. It is clear that more research related to anthropogenic influences along the Eerste/Kuils River catchment system are needed, specifically in monitoring monthly variations to better understand annual variation in several of the endpoints studied.
AFRIKAANSE OPSOMMING: Waterkwaliteit vorm die basis vir die evaluering en monitering van opvangsgebiede. Voortdurende stedelike ontwikkeling gee aanleiding tot ‘n toename in die voorkoms van besoedelstowwe in die natuurlike omgewing deur gelokaliseerde (punt) bronne (rioolwerke/industriële uitvloeisel) en/of nie gelokaliseerde (nie punt) bronne (vloed uitlaat/huishoudelike uitvloeisels) van besoedeling. Dit het onlangs aan die lig gekom dat van hierdie chemise besoedelstowwe subtiel die endokriene sisteem versteur en so aanleiding gee tot gesondheidsprobleme in terme van die voortplantingsisteem en tiroied sisteem (groei en ontwikkeling) by diere en moontlik ook die mens. Daar is beperkte gebruik van biologiese toetse wat inligting verskaf oor sitotoksiteit, inflammatoriese aktiwiteit en endokriene versteuring. Die doel van hierdie studie was dus om van hierdie biologiese toetse, wat normaalweg nie deel uitmaak van die roetine munisipale (Stad van Kaapstad) opvangsgebied monitering nie, gebruik te maak. Die Eerste-, Kuilsrivier, Wes Kaap, onder beheer van Stad Kaapstad is gekies en alhoewel die opvangsgebied nie water bydra tot drinkwaterbronne nie, word die opvangsgebied beïnvloed deur verskeie mensgemaakte bronne van besoedeling (afloop vanuit omliggende lanbougebiede). Binne die kort tydsraamwerk van die projek (ses maande) is besluit om twee maande, Julie (volg ‘n droë somer en herfs seisoen) en Oktober (volg ‘n nat winter en vroeë lente seisoen) vir water kwaliteit monitering te kies. Ruimtelike variasie langs die loop van die opvangsgebied is ingesluit deur moniteringspunte (n=10), met in ag name van die potensiële besoedelingsbronne. Spesifieke doelwitte van die projek sluit in: Eerstens (Hoofstuk 2), om die in vitro biotoetse, laktaat hidrolise (LDH) vir sitotoksisiteit, proinflammatoriese hormoon Interleuikin-6 (IL-6) vir inflammatoriese aktiwiteit, vrygestel deur menslike bloedselle en ‘n Salmonella ELISA vir ontlasting besoedeling saam met bestaande chemise en biologiese (hoofsaaklik migrobiologiese) veranderlikes te gebruik. Die studie het getoon dat beduidende variasie in alle mikrobiologiese toetse asook IL-6 vrystelling en Salmonella voorkoms bestaan het tussen versamelpunte. Maandelikse variasie in sekere van die veranderlikes het ook voorgekom. Tweedens (Hoofstuk 3), is twee biotoetse wat die dooiervoorloperproteïen, vitellogeen (Vtg) as eindpunt gebruik geimplimenteer in a) ‘n in vitro Xenopus laevis lewersnit biotoets (vyf dag blootstelling) en b) ‘n in vivo Zebravis (Danio rerio) biotoets (sewe dag blootstelling) om estrogenisiteit in die Eerste-, Kuilsrivier opvangsgebied te evalueer. Alhoewel, die estrogeen behandelde positiewe kontrole water monsters Vtg produksie veroorsaak het in beide die in vitro lewer-kulture en in vivo vistoets, is geen dramatiess estrogeniese aktiwiteit by enige van die moniteringspunte gevind nie. Derdens (Hoofstuk 4), is ‘n biotoets wat die tiroïedbeheerde metamorfose in Xenopus laevis paddavisse gebruik om effekte op die tiroïedsisteem te evalueer. Die differensiële stimulering (versnelling), in vergelyking met ‘n negatiewe kontrole watermonster, van die tiroied sisteem is by twee moniteringspunte in die opvangsgebied waargeneem. Alhoewel die praktiese implementering van die paddavis semi-statiese (water word gereeld vervang) biotoets arbeidsintensief is, het alle bykomstige biotoetse waardevolle bykomstige inligting oor water kwaliteit verskaf wat ‘n belangrike bydrae tot ingeligte bestuursbesluite kan verleen. Dit is duidelik dat meer navorsing int verband met die menslike invloed langs die Eerste-, Kuilsrivier opvangsgebied nodig is, veral met maandlikse monitering vir seisoenale veranderinge.

Le diabète mellitus, également désigné comme la maladie du siècle, est une pathologie mortelle qui affecte le système endocrinien. Les mécanismes liés à la rupture de la boucle de rétroaction, qui régule le métabolisme et induit le diabète, ne sont pas entièrement connus. La compréhension des mécanismes d'action de l'insuline est donc essentielle pour le développement de stratégies thérapeutiques efficaces afin du lutter contre cette maladie. Par conséquent, il est impératif de trouver un modèle robuste et fiable, capable de surmonter les limites de la culture cellulaire traditionnelle en 2D et de l'expérimentation animale, pour la recherche sur le diabète. L'objectif de cette thèse est de développer un nouveau modèle de co‐culture foie‐pancréas en utilisant des systèmes microphysiologiques avancés (MPs) afin d’aborder plus efficacement le mécanisme impliqué dans la régulation endocrinienne hépatique et pancréatique. Ce travail met en évidence la capacité des systèmes multi‐organes sur puce qui combinent la compartimentation avancée des cellules en 3D, la microfluidique et la technologie des cellules souches pluripotentes induites (iPSC), pour atteindre une complexité biologique élevée et des fonctions rarement reproduites par une seule de ces technologies d’ingénierie tissulaire
Diabetes mellitus (DM) or the so called disease of the century is a life threatening dysfunction that affects the endocrine system. The mechanisms underlying the break in the feedback loop that regulates the metabolism and the consequent diabetes induction are not fully known. Understanding the mechanisms of insulin action is therefore crucial for the further development of effective therapeutic strategies to combat DM. Accordingly, it is imperative to find a robust and reliable model for diabetes research able to overcome the limitations of traditional 2D in vitro cell culture and animal experimentation. The aim of this thesis is to develop a new liver‐pancreas co‐culture model using advanced microphysiological systems (MPs) to tackle more effectively the mechanism involving the hepatic and pancreatic endocrine regulation. This work highlights the power of multi organ‐on‐chip systems that combines the advanced 3D‐cell compartmentalization, microfluidics and induced pluripotent stem cells (iPSC) technology to achieve a high biological complexity and functions that are rarely reproduced by only one of these tissue engineering technologies

Etude réalisée pendant la gestation chez la rate rendue diabétique par injection de streptozotocine, dans le but d'identifier quelques facteurs impliqués dans le développement de l'hypotrophie foetale: sont évalués le rôle des perturbations hémodynamiques utéroplacentaires, et l'hypertonie du myomètre, avec les facteurs neuro-endocriniens impliqués dans ces modifications

Orientação: João Martins ; coorientação: Pedro Almeida
Este relatório de estágio foi realizado no âmbito da conclusão do Mestrado Integrado em Medicina Veterinária da Universidade Lusófona de Humanidades e Tecnologias e é composto por duas partes. A primeira tem por base descrever as atividades realizadas no estágio curricular realizado na clínica veterinária VetSesimbra e a segunda o desenvolvimento da endocrinopatia hiperadrenocorticismo, realizando sempre que possível o paralelismo entre cães e gatos. Esta dissertação é composta por uma revisão bibliográfica aprofundada acerca do tema e a apresentação e discussão de quatro casos clínicos – três em cães e um num gato – todos observados e acompanhados no decorrer do estágio curricular. O hiperadrenocorticismo é bastante frequente em cães e raro em gatos e é caracterizado pelo aumento do cortisol sérico nos animais afetados. Este aumento pode ser motivado pela administração exógena de glucocorticoides, por um excesso de produção de corticotrofina pela hipófise ou por uma neoplasia adrenocortical. Os sinais clínicos englobam polidipsia, poliúria, polifagia, alopecia simétrica bilateral, abdómen pendular e hepatomegalia e provocam degradação da qualidade de vida do animal. O diagnóstico de HAC baseia-se na anamnese, no exame clínico, nas análises bioquímicas, testes endócrinos e exames imagiológicos. O tratamento pode ser cirúrgico ou médico, sendo o fármaco mais utilizado o trilostano. Por fim, o prognóstico, difere consoante a etiologia, o tratamento utilizado e a existência de patologias concomitantes ou complicações associadas.
This internship report was written in order to complete the Integrated Master's Degree in Veterinary Medicine of Universidade Lusófona de Humanidades e Tecnologias and it consists of two parts. The first is based on the activities carried out during the curricular internship performed at the veterinary clinic VetSesimbra and the second on the development of hyperadrenocorticism endocrinopathy in dogs and cats. This dissertation is composed of an in-depth bibliographic review on the subject and the presentation and discussion of four clinical cases - three in dogs and one in a cat - all observed and monitored during the curricular internship. Hyperadrenocorticism is quite frequent in dogs and rare in cats and is due to the increase in serum cortisol in affected animals. This increase may be caused by the exogenous administration of glucocorticoids, an excess of corticotrophin production by the pituitary, or an adrenocortical neoplasm. Clinical signs include polydipsia, polyuria, polyphagia, bilateral symmetrical alopecia, pendular abdomen and hepatomegaly, and cause degradation of the animal's quality of life. The diagnosis is based on anamnesis, clinical examination, biochemical analyses, endocrine tests and imaging exams. The treatment can be surgical or medical, being the trilostane the most used drug. Finally, the prognosis differs depending on the etiology, the applied treatment and the existence of concomitant pathologies or associated complications.

Ο φαρμακευτικός κλάδος μπορεί να επωφεληθεί, με την βοήθεια της σημερινής τεχνολογίας των ηλεκτρονικών υπολογιστών και συγκεκριμένα των βάσεων δεδομένων. Σκοπός της διπλωματικής εργασίας είναι η κάλυψη των αναγκών της Ελληνικής συνταγογραφίας των φαρμάκων. Συγκεκριμένα οι φαρμακευτικές βάσεις δεδομένων, που ήδη υπάρχουν, δεν χρησιμοποιούν την τεχνολογία αναζήτησης των αλληλεπιδράσεων των φαρμάκων. Για το σκοπό αυτό σχεδιάστηκε και υλοποιήθηκε ένα μοντέλο βάσεων δεδομένων, που υποστηρίζει πλήρως τις αλληλεπιδράσεις των φαρμάκων, σύμφωνα με τις οδηγίες του εθνικού συνταγολογίου φαρμάκων (Ε.Ο.Φ). Έπειτα έγινε η εισαγωγή των φαρμάκων και δοκιμάστηκε η ακεραιότητα του συστήματος για την επιτυχή λειτουργία των αλληλεπιδράσεων μεταξύ φαρμάκων, καθώς μια εσφαλμένη αλληλεπίδραση μπορεί να αποβεί μοιραία για κάποιους ασθενείς. Τέλος αυτή η διπλωματική εργασία μπορεί να χρησιμοποιηθεί από φαρμακοποιούς και ιατρούς για την ασφαλέστερη και αποτελεσματικότερη συνταγογράφηση.
The pharmaceutical branch can profit, with the help of current technology of computers and concretely with the use of data bases. The purpose of this diplomatic work is to cover the needs of the Greek prescription on medicines. Particularly the pharmaceutical data bases, that already exist, do not use the search technology of interactions in medicines. For this aim it was drawn and implemented a model of data bases, that completely supports the interactions of medicines, according to the directives of national organization for medicines (N.O.M). The next step was the insertion of medicines and test the integrity of system for the successful operation of interactions between medicines, while a mistaken in interaction it can be turns out fatal for certain patients. Finally this diplomatic work can be used from pharmacists and doctors for secure and more effective prescription.

碩士
元培科學技術學院
影像醫學研究所
95
Recently, children with development disease were discovered in a lot of pediatric departments. General clinical diagnosis of this kind disease included height, weight, blood, urine and bone age measurements. Special examinations included chromosome examination, radioimmunoassay, computed tomography and magnetic resonance imaging. When growth hormone was insufficient, the last two methods could be used for the differentiation of brain structure abnormality and brain tumor. Generally, pituitary gland was evaluated by the changes of shape, size and height in different sections of image. However, computed tomography usually has scruple with the radiation dose, especially for children. In addition, two-dimensional images always could not provide effective information sufficiently. Therefore, the non-invasive magnetic resonance imaging method was employed to measure the pituitary volumes of children in Taiwan area. From our study, the comparison exterior between normal with exceptional pituitary gland in Taiwan area could be built. Furthermore, the information could also be provided as the reference of clinical diagnosis in pediatrics department. Children under the age of 14 and undergoing brain MRI for endocrines development disease or for other diagnostic purpose were our major subjects. Sixty-three children were eligible (mean age, 5.8 years). Forty images were collected by modern MRI Spoiled Gradient-recalled Acquisition in the Steady-state (SPGR) technique for each subject. Direct pituitary volumes were then measured after multi-plane reconstruction (MPR) for each image. In total, forty images of axial and forty images of saggittal plane were reconstructed separately for each subject. Due to the irregular three dimensional shape of pituitary gland, its volume measurement is a big challenge. In order to improve the accuracy and avoid errors, the area of each image was measured six times and then the average value was calculated. Finally, the volume of each plane was obtained by summarizing all averaged images of that plane. To confirm the trusty of our results, the volumes and errors of a 500 mm3 and a 250 mm3 image test phantoms were measured for calibration. The volume of pituitary gland for each subject could then be corrected by statistical method. Our results indicated that the average volumes of sexual precocity children were higher than normal children (p < 0.05), which had statistically significance. The average volumes of development delay children were smaller than normal children (p > 0.05), however did not have statistically significance. We thought the reason could be as following: the disease of development delay always found in much younger children, who usually have smaller volume of pituitary gland, and therefore generated more deviations from the averaged data of normal subjects.