Estudos em modelos animais transgênicos possibilitaram o conhecimento de parte dos genes envolvidos na embriogênese hipofisária e da etiologia genética do hipopituitarismo em humanos. Entretanto, a etiologia da maior parte dos casos de hipopituitarismo congênito, principalmente os associados à neuro-hipófise ectópica (NE), ainda é pouco definida. Mutações no gene PROP1 são a causa genética mais comum de hipopituitarismo descritas até o momento, mas estão sempre associadas à neuro-hipófise tópica. Estudos destinados a esclarecer o mecanismo molecular da mutação do gene Prop1 em camundongos demonstraram a participação da via de sinalização Notch e de seus componentes, dentre eles, o gene Hes1. O HES1 é um gene que codifica um fator de transcrição que participa de estágios precoces do desenvolvimento hipofisário e está envolvido com a morfogênese da neuro-hipófise. A avaliação do camundongo com nocaute em homozigose deste gene acarreta uma hipoplasia da adeno-hipófise e ausência da neuro-hipófise; e sua expressão constitutiva está associada ao hipopituitarismo. Como a NE é um achado comum no hipopituitarismo congênito e o gene HES1 pode estar relacionado a sua fisiopatologia, a região codificadora do gene HES1 foi avaliada em 192 pacientes com hipopituitarismo congênito. A variante alélica c.578G > A (p.G193D) em heterozigose foi encontrada em um paciente com hipopituitarismo congênito associado à NE. A avaliação da predição in silico do efeito funcional da variante pela ferramenta MutationTaster sugere que a troca do aminoácido glicina, altamente conservado entre os mamíferos, por ácido aspártico, seja deletéria. No estudo da segregação familiar, quatro irmãos aparentemente normais apresentam a mesma variante, sendo que dois deles possuem alterações discretas na imagem da hipófise. Em conclusão, esta é uma nova variante alélica descrita no gene HES1, ausente em grandes bancos de dados e controles saudáveis da população brasileira, mas presente em irmãos não afetados. Estudos funcionais in vitro são necessários para esclarecer o efeito biológico desta variante. Um padrão de herança complexo com penetrância incompleta é possível e já descrito em outros genes associados ao hipopituitarismo. Na tentativa de elucidar a causa genética do hipopituitarismo neste caso, o material genético deste paciente e de seus familiares foram submetidos ao sequenciamento do exoma, mas os resultados estão inconclusivos até o momento / Studies of transgenic animal models have allowed for the discovery of genes involved in human pituitary embryogenesis and the genetic etiology of hypopituitarism. However, the genetic causes of most cases of congenital hypopituitarism, especially those associated with an ectopic posterior pituitary, remain poorly defined. Mutations in the gene PROP1 are the most common genetic causes of hypopituitarism described to date, and are always associated with an ectopic posterior pituitary. Studies to elucidate the molecular mechanisms of Prop1 mutations in mice have demonstrated the involvement of the Notch signaling pathway, including its downstream target Hes1. The HES1 gene encodes a transcription factor that participates in early stages of pituitary development and is involved in posterior pituitary morphogenesis. Hes1 knockout mice exhibit a hypoplastic anterior pituitary and absence of a posterior pituitary. Conversely, constitutive expression of Hes1 is associated with hypopituitarism. Since an ectopic posterior pituitary is commonly found in congenital hypopituitarism and the HES1 gene may be related to its pathophysiology, the coding region of gene HES1 was screened in 192 patients with congenital hypopituitarism. A heterozygous allelic variant c.578G >A (p.G193D) was identified in a patient with congenital hypopituitarism associated with an ectopic posterior pituitary. Assessment by MutationTaster, a bioinformatic tool for in silico prediction of functional effect of missense variants, suggests that substitution of glycine (a highly conserved amino acid in this position among mammals) for aspartic acid is deleterious. In the genetic study of family members, we identified four apparently normal siblings with the same variant, two of which have discrete changes in their pituitary MRI. In conclusion, we described a new allelic variant in the gene HES1, absent in large databases and healthy Brazilian controls, but present in the unaffected siblings. In vitro functional studies are needed to clarify the biological effect of this variant. A complex pattern of inheritance with incomplete penetrance is possible in this case, as it has already been described in other genes associated with hypopituitarism. In an attempt to elucidate the genetic cause of hypopituitarism in the family described, DNA samples of this patient and his family were submitted to exome sequencing, but results are inconclusive at this time

En esta Tesis se ha realizado análisis, diseño y caracterización de los amplificadores de bajoruido y banda ancha en tecnología de GaAs PHEMT con aplicación a los módulos posteriores delradiómetro del instrumento de baja frecuencia del satélite Planck. La Tesis se compone de las siguientes partes:- Introducción y estudio del funcionamiento del radiómetro del instrumento de baja frecuencia de Planck.- Diseño y caracterización de amplificadores de bajo ruido utilizando tecnología de GaAs. Se presentan diseños MMIC en la banda Ka y en la banda Q, y un diseño MIC en la banda Q.- Diseño y construcción de los módulos posteriores en las bandas de 30 y 44 GHz. Se presentan varios prototipos fabricados en ambas bandas, así como medidas de cada uno de los subsistemas que los forman.- Desarrollo de técnicas de medida para receptores de banda ancha con detección directa y su aplicación a la caracterización de los módulos posteriores, mostrando el funcionamiento de los prototipos representativos para las dos bandas de frecuencia.- Integración de los módulos posteriores con los módulos frontales y presentación de algunos de los resultados de medida de los radiómetros completos. / This Thesis deals with the analysis, design and characterization of broadband low noise amplifiersin GaAs PHEMT technology with application to the radiometer Back-End Modules for the Planck Low Frequency Instrument (LFI). The Thesis is composed of the next parts:- Introduction and study about the radiometer of the Planck low frequency instrument.- Design and characterization of low noise amplifiers using GaAs technology. Ka-band MMIC designs and Q-band MMIC and a MIC design are presented.- Design and assembly of the 30 and 44 GHz back-end modules. Several prototypes have been manufactured in both frequency bands and the most representative test results of each subsystem are presented.- Development of measurement techniques for broadband direct detection receivers and their application to the characterization of the back-end modules. Performance of representative prototypes in both frequency bands is included.- Integration of the back end modules and front end modules and significant results of the tests for a radiometer in each frequency band.

Σκοπός: Η προσέγγιση και η αντιμετώπιση των κακώσεων της Αυχενικής Μοίρας της Σπονδυλικής Στήλης (ΑΜΣΣ) εξακολουθεί και σήμερα να παρουσιάζει διαφορές μεταξύ των διαφόρων κέντρων. Κατά καιρούς μάλιστα έχουν προταθεί πλείστοι τρόποι αντιμετώπισης: από συντηρητική με κρανιακή έλξη μέχρι πολύ επιθετική χειρουργική αντιμετώπιση με συνδυασμένες πρόσθιες και οπίσθιες προσπελάσεις. Στόχος της διατριβής ήταν η παρουσίαση της χειρουργικής εμπειρίας της Ορθοπαιδικής Κλινικής του Πανεπιστημίου Πατρών στην αντιμετώπιση των ασθενών αυτών και η ανάλυση των επιπλοκών των διαφόρων μεθόδων σταθεροποίησης. Έγινε προσπάθεια να απαντηθούν τα ακόλουθα ερωτήματα: 1) Ποια πρέπει να είναι σήμερα η διαγνωστική προσέγγιση των ασθενών με κάκωση στην ΑΜΣΣ; 2) Ποιοι ασθενείς χρειάζονται σταθεροποίηση; 3) Τι είδους σταθεροποίηση και ποιες είναι οι επιπλοκές αυτής; Ακολούθως, ποιος πρέπει να είναι ο σύγχρονος αλγόριθμος προσέγγισης των ασθενών με κακώσεις στην ΑΜΣΣ; 4) Ποιες είναι οι μακροχρόνιες επιπτώσεις της σταθεροποίησης; Μεθοδολογία: Εκατόν-δώδεκα ασθενείς με ασταθείς κακώσεις στην ΑΜΣΣ υποβλήθηκαν στην Κλινική μας σε πρόσθια, οπίσθια σταθεροποίηση ή και στις δύο. Ένας ασθενής θεωρούνταν ότι είχε ασταθή κάκωση της ΑΜΣΣ εάν είχε 5 ή παραπάνω βαθμούς με βάση τα κριτήρια αστάθειας των White & Panjabi. Τουλάχιστον ένας χρόνος παρακολούθησης (follow-up) ήταν αναγκαίος για να ενταχθεί ένας ασθενής στη μελέτη, με αποτέλεσμα να επιλεγούν τελικά 97 ασθενείς. Εβδομηντατέσσερις ασθενείς υποβλήθηκαν σε αριστερή προσθιοπλάγια προσπέλαση [Ομάδα Α]. Σε 65 ασθενείς έγινε πρόσθια αποσυμπίεση και τοποθέτηση φλοιοσπογγώδους λαγονίου αυτομοσχεύματος και σταθεροποίηση με πλάκα και βίδες, είτε με πλάκα της AO/ASIF ή με πλάκα CSLP. Στους υπόλοιπους 9 ασθενείς, η αποκατάσταση της σπονδυλικής στήλης περιελάμβανε τη χρήση κλωβού πλέγματος τιτανίου στο οποίο τοποθετούνταν σπογγώδη αυτομοσχεύματα από την περιοχή της σωματεκτομής. Εικοσιτρείς ασθενείς υποβλήθηκαν σε οπίσθια σταθεροποίηση είτε με πλάκες πλαγίων ογκωμάτων (πλάκες Roy-Camille) (19 ασθενείς), ή με πολυαξονικές βίδες (4 ασθενείς) [Ομάδα Β]. Αποτελέσματα – Συμπεράσματα: 1) Εφόσον η κλινική εικόνα ενός ασθενούς με κάκωση στην ΑΜΣΣ επιβάλλει τη διενέργεια αξονικής τομογραφίας, η διερεύνηση μπορεί να γίνει με ασφάλεια με τη χρήση ενός σύγχρονου πολυτομικού αξονικού τομογράφου (MDCT) και μόνο, παραλείποντας τις απλές ακτινογραφίες. 2) Τα κριτήρια αστάθειας των White και Panjabi υπαγορεύουν μια ασταθή κάκωση στην ΑΜΣΣ, η οποία θα πρέπει να αντιμετωπίζεται κατά προτίμηση με χειρουργικό τρόπο. 3) Η στατιστική ανάλυση των αποτελεσμάτων δεν ανέδειξε στατιστικά σημαντικές διαφορές μεταξύ των 2 ομάδων σταθεροποίησης όσον αφορά τις κλινικά σημαντικές επιπλοκές (p=0.26). Ομοίως, οι κλινικά μη σημαντικές επιπλοκές, καθώς και το ποσοστό επανεγχειρήσεων δε διέφεραν στατιστικά μεταξύ των δύο ομάδων (p=0.245 και p=0.475 αντίστοιχα). Ωστόσο, η πρόσθια σταθεροποίηση παρουσιάζει σημαντικά πλεονεκτήματα, μπορεί να αντιμετωπίσει το σύνολο σχεδόν των κακώσεων της ΑΜΣΣ συμπεριλαμβανομένου των εξαρθρημάτων, και μόνο κατ’ εξαίρεση απαιτείται συμπληρωματική σταθεροποίηση. Επιπλέον, η εξέλιξη της μεθόδου βοήθησε στην εξάλειψη επιπλοκών που παρατηρούνταν με τα παλαιότερης τεχνολογίας υλικά. Ως εκ τούτου, οι τρέχουσες ενδείξεις για οπίσθια σταθεροποίηση είναι τα μη ανατασσόμενα εξαρθρήματα, οι κακώσεις πολλαπλών επιπέδων και οι ασθενείς με τραχειοστομία. 4) Η οστεοποίηση των παρακειμένων διαστημάτων στις κακώσεις της ΑΜΣΣ φαίνεται ότι έχει διαφορετική αιτιολογία από αυτή της αυχενικής σπονδύλωσης, μπορεί να εμφανιστεί πολύ πρώιμα στην μετεγχειρητική περίοδο και, ακόμα και όταν είναι εμφανής ακτινολογικά, σπανίως προκαλεί συμπτώματα. / Aim: The diagnostic approach and management of patients with cervical spine injuries differs among various centers. Conservative management with skeletal traction to aggressive surgical treatment with combined anterior and prosterior stabilization are within the possible alternatives. We aimed at presenting the experience from the surgical treatment of these patients gathered in the Department of Orthopaedic Surgery in the University Hospital of Patras. We further analyzed the complications associated with each approach and we tried to answer the following questions: 1) Which is the current diagnostic approach of patients with cervical spine injuries? 2) Which patients should be stabilized? 3) What type the stabilization should be and which are its complications? Moreover, which is the appropriate algorithm in the treatment of these patients? 4) Which are the long-term consequences of the stabilization? Materials & Methods: One hundred and twelve patients with unstable cervical spine injuries underwent anterior, posterior stabilization or both. A patient was considered to have an unstable injury if he had ≥ 5 points in the White and Panjabi checklist. At least one year of follow-up was necessary for a patient to be included in the study, which yielded a total of 97 patients. Seventy-four patients underwent a left-sided anterolateral approach [Group A]. Sixty-five of them had anterior decompression and iliac bone grafting. The remaining 9 patients underwent corpectomy and cervical spine reconstruction with titanium mesh cage, filled with morselized autograft from the corpectomy site. All these patients were instrumented using an anterior cervical plate. Twenty-three patients underwent posterior stabilization, either with lateral mass plates of Roy-Camille (19 patients), or polyaxial screws and rods (4 patients) [Group B], along with concomitant iliac bone autografting. Results – Conclusions: 1) If there is a need for computed tomography (CT) in a patient with cervical spine injury, the diagnostic work-up can be done with safety using only a modern multi-detector CT, obviating the need for plain radiographs. 2) The White and Panjabi criteria imply an unstable injury which should be preferentially stabilized by surgical means. 3) Statistical analysis of the clinically significant complications did not reveal significant difference between the posterior procedures and the anterior ones (p=0.26). Likewise, insignificant complications, as well as reoperation rates did not differ significantly among the two groups (p=0.245 and p=0.475 respectively). However, anterior stabilization for cervical spine injuries presents several advantages, can deal with almost all types of injuries and it only exceptionally requires supplemental stabilization. It should be also stressed that the advances in technology and metallurgy have eliminated the complications observed with older implants. Current indications for posterior stabilization are the irreducible dislocations, multilevel injuries and patients with tracheostomy. 4) Adjacent-level ossification in cervical spine injuries appears to be of different etiology than in cervical spondylosis, it may appear very early in the postoperative period and, even when it is evident radiographically, it very rarely (if ever) produces any symptoms.

No description available.

Les informations olfactives sont connues pour leur capacité à induire des comportements moteurs spécifiques. En dépit de nombreuses observations comportementales chez les vertébrés, on ne connaît toujours pas les mécanismes et les voies nerveuses qui sous-tendent ces phénomènes de transformation olfacto-locomotrices. Chez la lamproie, des travaux récents ont permis de décrire cette voie, et les mécanismes responsables de la transformation des entrées olfactives en activité locomotrice (Derjean et al., 2010). Cette voie prend origine dans la partie médiane du bulbe olfactif, et envoie des projections vers le tubercule postérieur, une région qui se trouve dans le diencéphale. De là, les neurones projettent directement vers la Région Locomotrice Mésencéphalique, connue pour envoyer des connexions vers les neurones réticulospinaux, et activer la locomotion. L’objectif de cette étude était d’établir si l’ensemble des neurones réticulospinaux répond aux stimulations olfactives. Pour ce faire, nous avons utilisé sur une préparation de cerveau isolé de lamproie des techniques d’électrophysiologie et d’imagerie calcique. La stimulation électrique des nerfs olfactifs, de la région médiane du bulbe olfactif ou du tubercule postérieur a provoqué une activation de toutes les cellules réticulospinales qui se retrouvent dans les quatre noyaux réticulaires (ARRN : Noyau Réticulaire Rhombencéphalique Antérieur; MRN : Noyau Réticulaire Mésencéphalique; MRRN : Noyau Réticulaire Rhombencéphalique Moyen; PRRN : Noyau Réticulaire Rhombencéphalique Postérieur). Seule la partie médiane du bulbe olfactif est impliquée dans le passage de l’information olfactive vers les neurones réticulospinaux. Nous avons aussi découvert que le blocage des récepteurs GABAergiques dans la partie médiane du bulbe olfactif augmentait les réponses olfactives de façon considérable dans les cellules réticulospinales. Nous avons montré ainsi qu’il existe un tonus inhibiteur impliqué dans la dépression modulatrice de la voie olfacto-locomotrice. Ce travail a permis de montrer que la stimulation des afférences sensorielles olfactives active simultanément l’ensemble des populations de neurones réticulospinaux qui commandent la locomotion. De plus, il existerait un tonus inhibiteur GABAergique, au niveau de la partie médiane du bulbe olfactif, responsable d’une dépression modulatrice dans la voie olfacto-locomotrice. / Olfactory inputs are known for their ability to induce specific motor behaviors. Despite numerous behavioral observations in vertebrates, the mechanisms and the neural pathways underlying the olfactory-locomotor transformation are still unknown. In lamprey, recent studies have described this pathway and the mechanism underlying the transformation of olfactory input into a locomotor activity (Derjean et al., 2010). This pathway originates in the medial part of the olfactory bulb, sends projections to the posterior tuberculum, a diencephalic region. From there, the neurons project directly to the mesencephalic locomotor region that is known to send projections to the reticulospinal neurons to activate locomotion. Using lamprey brain preparation, electrophysiology and calcium imaging, the aim of this study was to establish whether all reticulospinal neurons respond to olfactory stimuli. Electrical stimulation of the olfactory nerves, the medial part of the olfactory bulb or the posterior tuberculum activates all reticulospinal cells in the four reticular nuclei (ARRN: Anterior rhombencephalic reticular nucleus; MRN: middle mesencephalic reticular nucleus; MRRN: middle rhombencephalic reticular nucleus; PRRN: posterior rhombencephalic reticular nucleus). The medial part of the olfactory bulb is the only region that is implicated in transmitting the olfactory information to reticulospinal neurons. We also discovered that when blocking the GABAergic receptors in the medial part of the olfactory bulb, the reticulospinal neurons have a stronger response to olfactory stimulation. Thus we showed that a tonic inhibition is involved in the modulating depression of the olfacto-locomotor pathway. Altogether, this work shows that stimulation of the olfactory sensory inputs activates simultaneously the entire population of reticulospinal neurons that control locomotion. In addition, there is a GABAergic tonic inhibition at the level of the medial part of the olfactory bulb that causes a modulating depression in the olfacto-locomotor pathway.

No description available.

Introdução: Alterações em genes relacionados com a secreção de GH ou a organogênese hipofisária foram identificadas em pacientes com deficiência de hormônio do crescimento (DGH) congênita. Entretanto, poucos casos de DGH têm sua etiologia esclarecida. O GHRH é um candidato óbvio para explicar a deficiência isolada de GH (DIGH). Na literatura, os estudos de análise do GHRH não conseguiram identificar mutações, porém são antigos e utilizaram uma metodologia com limitações. A maioria dos pacientes com deficiência hipotálamo-hipofisária múltipla (DHHM) apresenta neuroipófise ectópica sugerindo a importância do estudo de genes que atuam no início do desenvolvimento hipofisário, com expressão inclusive no infundíbulo. O GLI2 é um fator de transcrição na sinalização Sonic Hedgehog, envolvido com o início da embriogênese hipofisária, expresso na bolsa de Rathke primordial e no diencéfalo ventral. Previamente, mutações no GLI2 foram encontradas em pacientes com holoprosencefalia, e também alterações hipofisárias. Objetivos: Analisar o GHRH em 151 pacientes com DIGH (42 brasileiros e 109 encaminhados de centros internacionais) e analisar o GLI2 em 180 pacientes brasileiros com DIGH ou DHHM por PCR e sequenciamento automático dos genes; e descrever o fenótipo dos pacientes com mutações identificadas. Resultados: No GHRH foram identificadas seis variantes em heterozigose com previsão benigna pelas análises in silico. A análise do GLI2 identificou três mutações novas em heterozigose com códon de parada prematuro (p.L788fsX794, p.L694fsX722 e p.E380X), e geração de proteínas truncadas, com perda do domínio responsável pela ativação transcricional. A mutação p.L788fsX794 foi identificada numa paciente com baixa estatura, polidactilia, epilepsia e hipoglicemias. Apresentava deficiência de GH, TSH, ACTH, prolactina, LH e FSH. Na investigação familiar foi diagnosticada DIGH em dois tios e DHHM numa prima. Estes familiares, além de sua mãe e outros parentes maternos também apresentaram a mutação e polidactilia. A mutação p.L694fsX722 foi identificada num menino com baixa estatura por deficiência de GH, além de lábio leporino e fenda palatina. Seu pai, embora saudável, também apresentou a mutação. A mutação p.E380X foi identificada numa lactente com retardo no desenvolvimento, hipoglicemias, poliúria e polidipsia. Apresentava deficiência de GH, ACTH, TSH e ADH. Sua mãe aparentemente normal também apresentou a mutação. Todos os pacientes com DGH e mutação no GLI2 apresentaram neuroipófise ectópica (não visualizada na paciente com p.E380X), adenoipófise hipoplásica e ausência de holoprosencefalia na ressonância magnética. Dezoito variantes não-sinônimas também foram identificadas no GLI2 em 24 pacientes. Dezesseis dessas variantes foram consideradas deletérias em pelo menos um programa de predição in silico e dez delas não foram encontradas em população controle. O fenótipo dos pacientes foi predominante de DHHM e com neuroipófise ectópica e sem holoprosencefalia. Variantes silenciosas, intrônicas e polimorfismos foram identificados no GLI2, mas aparentemente sem alteração funcional. Conclusão: Não identificamos mutação no GHRH e se realmente existe mutação neste gene como causa de DGH, deve ser muito rara. Variantes no GLI2 são frequentes (15%), indicando seu importante papel na etiologia da DGH congênita. Além disso, ampliamos o espectro fenotípico dos pacientes com mutações no GLI2, que foi caracterizado por DIGH ou DHHM, inclusive com diabetes insipidus, neuroipófise ectópica (maioria) e ausência de holoprosencefalia. Outras características observadas foram polidactilia, defeito de linha média facial e herança autossômica dominante com penetrância incompleta / Introduction: Alterations in genes related to GH secretion and pituitary organogenesis have been identified in patients with congenital GH deficiency (GHD). However, in only few cases of GHD the etiology has been established. GH-releasing hormone (GHRH) is an obvious candidate to explain isolated GH deficiency (IGHD). Previous reports in the literature did not identify mutations in GHRH, however, the methodology used was limited. Most patients with combined pituitary hormone deficiency (CPHD) have an ectopic posterior pituitary lobe (EPP) suggesting the study of genes involved in early pituitary development and also expressed in the infundibulum. GLI2 is a transcription factor in Sonic hedgehog signaling expressed in the primordial Rathkes pouch and ventral diencephalon during early pituitary development. Previously, GLI2 mutations were found in patients with holoprosencephaly and pituitary abnormalities. Aim: Analyse GHRH in 151 patients with IGHD (42 Brazilian and 101 from international centers) and GLI2 in 180 Brazilian patients with IGHD or CPHD by PCR and automatic sequencing, and describe the phenotype of patients with mutations. Results: In GHRH, six heterozygous variants that are benign according to in silico analysis were identified. GLI2 study revealed three novel heterozygous mutations leading to premature stop codons (p.L788fsX794, p.L694fsX722 e p.E380X) and truncated proteins, without the transcriptional activator domain. p.L788fsX794 was identified in a girl with short stature, polydactyly, epilepsy and hypoglycemia. She had GH, TSH, ACTH, prolactina, LH and FSH deficiencies. Two uncles had IGHD and one cousin CPHD. These relatives, the mother and other maternal relatives had polydactyly and carried the mutation. p.L694fsX722 was identified in a boy with short stature due to GHD who also had cleft lip and palate. His healthy father also carried the mutation. p.E380X was identified in an infant with delayed development, hypoglycemia, polyuria and polydipsia. She had GH, ACTH, TSH and ADH deficiencies. Her apparently normal mother also carried the mutation. All patients with GHD and GLI2 mutations had an EPP (not visualized in the patient with p.E380X), hypoplastic anterior pituitary lobe and absence of holoprosencephaly on MRI. Eighteen non-synonymous variants in GLI2 were identified in 24 patients. Sixteen of these were considered deleterious in at least one in silico prediction program and ten of these were not found in the control population. The phenotype was mainly of CPHD and EPP without holoprosencephaly. Several synonymous and intronic GLI2 variants and polymorphisms apparently without functional consequences were identified. Conclusions: No mutations in GHRH were identified and if mutations in this gene exist as a cause of IGHD, they must be very rare. Variants in GLI2 are frequent (15%) indicating its important role in the etiology of GHD. Furthermore, we expanded the clinical spectrum of patients with GLI2 mutations characterized by IGHD or CPHD including diabetes insipidus, ectopic posterior pituitary lobe (in most patients) and absence of holoprosencephaly. Additional features were polydactyly and midline facial defects and the inheritance was autosomal dominant with incomplete penetrance

Estudos em modelos animais transgênicos possibilitaram o conhecimento de parte dos genes envolvidos na embriogênese hipofisária e da etiologia genética do hipopituitarismo em humanos. Entretanto, a etiologia da maior parte dos casos de hipopituitarismo congênito, principalmente os associados à neuro-hipófise ectópica (NE), ainda é pouco definida. Mutações no gene PROP1 são a causa genética mais comum de hipopituitarismo descritas até o momento, mas estão sempre associadas à neuro-hipófise tópica. Estudos destinados a esclarecer o mecanismo molecular da mutação do gene Prop1 em camundongos demonstraram a participação da via de sinalização Notch e de seus componentes, dentre eles, o gene Hes1. O HES1 é um gene que codifica um fator de transcrição que participa de estágios precoces do desenvolvimento hipofisário e está envolvido com a morfogênese da neuro-hipófise. A avaliação do camundongo com nocaute em homozigose deste gene acarreta uma hipoplasia da adeno-hipófise e ausência da neuro-hipófise; e sua expressão constitutiva está associada ao hipopituitarismo. Como a NE é um achado comum no hipopituitarismo congênito e o gene HES1 pode estar relacionado a sua fisiopatologia, a região codificadora do gene HES1 foi avaliada em 192 pacientes com hipopituitarismo congênito. A variante alélica c.578G > A (p.G193D) em heterozigose foi encontrada em um paciente com hipopituitarismo congênito associado à NE. A avaliação da predição in silico do efeito funcional da variante pela ferramenta MutationTaster sugere que a troca do aminoácido glicina, altamente conservado entre os mamíferos, por ácido aspártico, seja deletéria. No estudo da segregação familiar, quatro irmãos aparentemente normais apresentam a mesma variante, sendo que dois deles possuem alterações discretas na imagem da hipófise. Em conclusão, esta é uma nova variante alélica descrita no gene HES1, ausente em grandes bancos de dados e controles saudáveis da população brasileira, mas presente em irmãos não afetados. Estudos funcionais in vitro são necessários para esclarecer o efeito biológico desta variante. Um padrão de herança complexo com penetrância incompleta é possível e já descrito em outros genes associados ao hipopituitarismo. Na tentativa de elucidar a causa genética do hipopituitarismo neste caso, o material genético deste paciente e de seus familiares foram submetidos ao sequenciamento do exoma, mas os resultados estão inconclusivos até o momento / Studies of transgenic animal models have allowed for the discovery of genes involved in human pituitary embryogenesis and the genetic etiology of hypopituitarism. However, the genetic causes of most cases of congenital hypopituitarism, especially those associated with an ectopic posterior pituitary, remain poorly defined. Mutations in the gene PROP1 are the most common genetic causes of hypopituitarism described to date, and are always associated with an ectopic posterior pituitary. Studies to elucidate the molecular mechanisms of Prop1 mutations in mice have demonstrated the involvement of the Notch signaling pathway, including its downstream target Hes1. The HES1 gene encodes a transcription factor that participates in early stages of pituitary development and is involved in posterior pituitary morphogenesis. Hes1 knockout mice exhibit a hypoplastic anterior pituitary and absence of a posterior pituitary. Conversely, constitutive expression of Hes1 is associated with hypopituitarism. Since an ectopic posterior pituitary is commonly found in congenital hypopituitarism and the HES1 gene may be related to its pathophysiology, the coding region of gene HES1 was screened in 192 patients with congenital hypopituitarism. A heterozygous allelic variant c.578G >A (p.G193D) was identified in a patient with congenital hypopituitarism associated with an ectopic posterior pituitary. Assessment by MutationTaster, a bioinformatic tool for in silico prediction of functional effect of missense variants, suggests that substitution of glycine (a highly conserved amino acid in this position among mammals) for aspartic acid is deleterious. In the genetic study of family members, we identified four apparently normal siblings with the same variant, two of which have discrete changes in their pituitary MRI. In conclusion, we described a new allelic variant in the gene HES1, absent in large databases and healthy Brazilian controls, but present in the unaffected siblings. In vitro functional studies are needed to clarify the biological effect of this variant. A complex pattern of inheritance with incomplete penetrance is possible in this case, as it has already been described in other genes associated with hypopituitarism. In an attempt to elucidate the genetic cause of hypopituitarism in the family described, DNA samples of this patient and his family were submitted to exome sequencing, but results are inconclusive at this time

Abstract Neuropsychiatric illnesses usually become overtly manifest in adolescence and early adulthood. A critical long-term aim is to be able to prevent the development of such illnesses, which requires instruments to identify subjects at high risk of illness and to offer them effective interventions. There is an indisputable need for more sophisticated methods to enable more precise detection of adolescents and young adults who are at high risk of developing psychosis. Abnormal function in brain networks has been reported in people with schizophrenia and other psychotic disorders. Similar abnormalities have been found also in people at risk for developing psychosis, but it is not known whether this applies also to spontaneous resting state activity in young people with a familial risk for psychosis. We conducted resting-state functional MRI (R-fMRI) in 72 (29 male) young adults with a history of psychosis in one or both parents (FR) but without psychosis themselves, and 72 (29 male) similarly healthy control subjects without familial risk for psychosis. Both groups in the Oulu Brain and Mind study were drawn from the Northern Finland Birth Cohort 1986. All volunteers were 20–25 years old. Parental psychosis was established using the Care Register for Health Care. R-fMRI data was pre-processed using independent component analysis (ICA). A dual regression technique was used to detect between-group differences with p < 0.05 threshold corrected for multiple comparisons at voxel level. FR subjects demonstrated significantly decreased activity compared to control subjects in the default mode network and in the central executive network and increased activity in the cerebellum. The findings clarify previously controversial literature on the subject. The finding suggests that abnormal activity in these brain networks in rest may be associated with increased vulnerability to psychosis. The findings maybe helpful in developing more precise methods for detecting young people at highest risk for developing psychosis. / Tiivistelmä Psykoottisiin häiriöihin sairastutaan yleensä nuoruudessa tai varhaisaikuisuudessa. Psykoositutkimuksen tavoitteena on löytää uusia menetelmiä, joiden avulla kyettäisiin tunnistamaan suurimmassa psykoosiriskissä olevat nuoret, jotta heille voitaisiin tarjota sairautta ennaltaehkäiseviä hoitokeinoja. Skitsofreniaan ja muihin psykoottisiin häiriöihin sairastuneilla on havaittu aivotoiminnan poikkeavuuksia. Samankaltaisia aivotoiminnan poikkeavuuksia on havaittu myös nuorilla, jotka ovat vaarassa sairastua psykoosiin. Toistaiseksi on ollut epäselvää, onko psykoosiin sairastuneiden henkilöiden lapsilla aivohermoverkkojen toiminnan poikkeavuuksia lepotilassa. Suoritimme aivojen lepotilan MRI-tutkimuksen (R-fMRI) 72:lle (29 miestä) nuorelle aikuiselle, joiden jompikumpi vanhempi oli sairastunut psykoosin sekä 72:lle (29 miestä) nuorelle aikuiselle, joiden vanhemmat eivät olleet sairastaneet psykoosia. Molemmat tutkimusryhmät tässä Oulu Brain and Mind -tutkimuksessa olivat Pohjois-Suomen 1986 syntymäkohortin jäseniä. Tutkittavat olivat 20–25 vuoden iässä. Lepotilan toiminnallinen magneettikuvaus suoritettiin 1.5 Teslan Siemensin magneettikuvantamislaitteella. Tutkimuskohteiksi valittiin lepotilan toiminnallinen aivohermoverkko, toiminnan ohjauksesta vastaava aivohermoverkko ja pikkuaivot. Kuvantamisdataan sovellettiin itsenäisten komponenttien analyysia aivohermoverkkojen määrittämistä varten. Ryhmien välisen eron havaitsemiseen käytettiin ei-parametristä permutaatiotestiä, joka kynnystettiin tilastollisesti merkitsevään tasoon (p < 0.05). Lepotilan oletushermoverkossa ja toiminnanohjauksesta vastaavassa aivohermoverkoissa havaittiin vähäisempää aktiivisuutta ja pikkuaivoissa kohonnutta aktiivisuutta perinnöllisessä psykoosiriskissä olevilla nuorilla aikuisilla verrattuna verrokkeihin. Tutkimustulokset selkeyttivät aiempaa ristiriitaista kirjallisuutta tutkimusaiheesta. Tutkimuksessa havaittujen aivoalueiden poikkeava toiminta lepotilassa voi liittyä kohonneeseen psykoosin puhkeamisriskiin. Tutkimuslöydösten avulla voidaan todennäköisesti edesauttaa parempien kuvantamismenetelmien kehittämistä suurimmassa psykoosiriskissä olevien nuorten tunnistamiseen.

Cette thèse aborde le processus ponctuel déterminantal, un modèle probabiliste qui capture la répulsion entre les points d’un certain espace. Celle-ci est déterminée par une matrice de similarité, la matrice noyau du processus, qui spécifie quels points sont les plus similaires et donc moins susceptibles de figurer dans un même sous-ensemble. Contrairement à la sélection aléatoire uniforme, ce processus ponctuel privilégie les sous-ensembles qui contiennent des points diversifiés et hétérogènes. La notion de diversité acquiert une importante grandissante au sein de sciences comme la médecine, la sociologie, les sciences forensiques et les sciences comportementales. Le processus ponctuel déterminantal offre donc une alternative aux traditionnelles méthodes d’échantillonnage en tenant compte de la diversité des éléments choisis. Actuellement, il est déjà très utilisé en apprentissage automatique comme modèle de sélection de sous-ensembles. Son application en statistique est illustrée par trois articles. Le premier article aborde le partitionnement de données effectué par un algorithme répété un grand nombre de fois sur les mêmes données, le partitionnement par consensus. On montre qu’en utilisant le processus ponctuel déterminantal pour sélectionner les points initiaux de l’algorithme, la partition de données finale a une qualité supérieure à celle que l’on obtient en sélectionnant les points de façon uniforme. Le deuxième article étend la méthodologie du premier article aux données ayant un grand nombre d’observations. Ce cas impose un effort computationnel additionnel, étant donné que la sélection de points par le processus ponctuel déterminantal passe par la décomposition spectrale de la matrice de similarité qui, dans ce cas-ci, est de grande taille. On présente deux approches différentes pour résoudre ce problème. On montre que les résultats obtenus par ces deux approches sont meilleurs que ceux obtenus avec un partitionnement de données basé sur une sélection uniforme de points. Le troisième article présente le problème de sélection de variables en régression linéaire et logistique face à un nombre élevé de covariables par une approche bayésienne. La sélection de variables est faite en recourant aux méthodes de Monte Carlo par chaînes de Markov, en utilisant l’algorithme de Metropolis-Hastings. On montre qu’en choisissant le processus ponctuel déterminantal comme loi a priori de l’espace des modèles, le sous-ensemble final de variables est meilleur que celui que l’on obtient avec une loi a priori uniforme. / This thesis presents the determinantal point process, a probabilistic model that captures repulsion between points of a certain space. This repulsion is encompassed by a similarity matrix, the kernel matrix, which selects which points are more similar and then less likely to appear in the same subset. This point process gives more weight to subsets characterized by a larger diversity of its elements, which is not the case with the traditional uniform random sampling. Diversity has become a key concept in domains such as medicine, sociology, forensic sciences and behavioral sciences. The determinantal point process is considered a promising alternative to traditional sampling methods, since it takes into account the diversity of selected elements. It is already actively used in machine learning as a subset selection method. Its application in statistics is illustrated with three papers. The first paper presents the consensus clustering, which consists in running a clustering algorithm on the same data, a large number of times. To sample the initials points of the algorithm, we propose the determinantal point process as a sampling method instead of a uniform random sampling and show that the former option produces better clustering results. The second paper extends the methodology developed in the first paper to large-data. Such datasets impose a computational burden since sampling with the determinantal point process is based on the spectral decomposition of the large kernel matrix. We introduce two methods to deal with this issue. These methods also produce better clustering results than consensus clustering based on a uniform sampling of initial points. The third paper addresses the problem of variable selection for the linear model and the logistic regression, when the number of predictors is large. A Bayesian approach is adopted, using Markov Chain Monte Carlo methods with Metropolis-Hasting algorithm. We show that setting the determinantal point process as the prior distribution for the model space selects a better final model than the model selected by a uniform prior on the model space.