Diagnostic value of H3F3A mutations in giant cell tumour of bone compared to osteoclast-rich mimics

Driver mutations in the two histone 3.3 (H3.3) genes, H3F3A and H3F3B, were recently identified by whole genome sequencing in 95% of chondroblastoma (CB) and by targeted gene sequencing in 92% of giant cell tumour of bone (GCT). Given the high prevalence of these driver mutations, it may be possible to utilise these alterations as diagnostic adjuncts in clinical practice. Here, we explored the spectrum of H3.3 mutations in a wide range and large number of bone tumours (n 5 412) to determine if these alterations could be used to distinguish GCT from other osteoclast-rich tumours such as aneurysmal bone cyst, nonossifying fibroma, giant cell granuloma, and osteoclast-rich malignant bone tumours and others. In addition, we explored the driver landscape of GCT through whole genome, exome and targeted sequencing (14 gene panel). We found that H3.3 mutations, namely mutations of glycine 34 in H3F3A, occur in 96% of GCT. We did not find additional driver mutations in GCT, including mutations in IDH1, IDH2, USP6, TP53. The genomes of GCT exhibited few somatic mutations, akin to the picture seen in CB. Overall our observations suggest that the presence of H3F3A p.Gly34 mutations does not entirely exclude malignancy in osteoclast-rich tumours. However, H3F3A p.Gly34 mutations appear to be an almost essential feature of GCT that will aid pathological evaluation of bone tumours, especially when confronted with small needle core biopsies. In the absence of H3F3A p.Gly34 mutations, a diagnosis of GCT should be made with caution.

Distinct H3F3A and H3F3B driver mutations define chondroblastoma and giant cell tumor of bone

It is recognized that some mutated cancer genes contribute to the development of many cancer types, whereas others are cancer type specific. For genes that are mutated in multiple cancer classes, mutations are usually similar in the different affected cancer types. Here, however, we report exquisite tumor type specificity for different histone H3.3 driver alterations. In 73 of 77 cases of chondroblastoma (95%), we found p.Lys36Met alterations predominantly encoded in H3F3B, which is one of two genes for histone H3.3. In contrast, in 92% (49/53) of giant cell tumors of bone, we found histone H3.3 alterations exclusively in H3F3A, leading to p.Gly34Trp or, in one case, p.Gly34Leu alterations. The mutations were restricted to the stromal cell population and were not detected in osteoclasts or their precursors. In the context of previously reported H3F3A mutations encoding p.Lys27Met and p.Gly34Arg or p.Gly34Val alterations in childhood brain tumors, a remarkable picture of tumor type specificity for histone H3.3 driver alterations emerges, indicating that histone H3.3 residues, mutations and genes have distinct functions.

Estudo da divergência entre duas vias que curvam em sentidos contrários

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Civil na Área de Especialização de Vias de Comunicação e Transportes

Grande Queimado numa Unidade de Cuidados Intensivos Pediátricos – Experiência de 20 Anos

Introdução: A abordagem inicial do grande queimado até à sua estabilização hemodinâmica e hidroeletrolítica é fundamental para diminuir a morbimortalidade. Material e Métodos: Estudo retrospectivo, descritivo e analítico, de todos os internamentos por queimadura numa Unidade de Cuidados Intensivos Pediátricos durante o período de 20 anos (Abril/1991 a Dezembro/2010). Avaliaram -se parâmetros nosodemográficos, agente causal, gravidade e extensão da queimadura, procedimentos, terapêutica, complicações e resultados. Resultados: Ocorreram 137 internamentos por queimadura correspondentes a 123 doentes e a 1,8% do total de internamentos na UCIP. A mediana de idade foi 3,6 anos e 62,4% era do sexo masculino. Verificou -se maior incidência em Agosto (13,0%). Foram agentes da queimadura: líquido fervente (38,1%), fogo (38,1%) e eletricidade (23,9%). A mediana da superfície corporal queimada foi de 30% (0,5 -92,0%), com queimaduras do terceiro grau em 59,0% dos doentes. Necessitaram de ventilação mecânica 45,5% e de cateter venoso central 64,2% dos doentes. As complicações incluíram: sépsis (29,2%), falência respiratória (21,1%), falência cardiovascular (16,5%) e falência multiorgânica (18,8%). Verificou -se melhoria em 88,6% dos casos e ocorreram 10 óbitos (8,1%), nove dos quais nos primeiros 10 anos do estudo e nove devido a causa infeciosa. No entanto, o score avaliador do risco de mortalidade (PRISM), índice de intervenção terapêutica (TISS) e o risco de probabilidade de morte (RPM) foram mais elevados no segundo decénio. Conclusões: Nos últimos anos do estudo, apesar do maior número de admissões e da sua maior gravidade, verificou -se uma diminuição do número de mortes, o que poderá dever-se à melhoria dos cuidados prestados.

Brotherly love and filial obedience: the commemorative programme of the Avis princes at Santa Maria da Vitória, Batalha

This dissertation focuses on a rare 15th century commemorative programme that has thus far received little scholarly attention: the collective monument erected in the Founder’s Chapel, at the Monastery of Santa Maria da Vitória, Batalha, to house the remains of four Avis princes, members of what would become known as ‘the Illustrious Generation’. A patron is proposed for the commission of this erudite monument - the princes’ eldest brother, king Duarte I - arguing its integration into a broader propaganda programme to glorify the memory of the Avis dynasty founder, king João I. The dissertation then proceeds to discuss various highly innovative features of the monument, such as its pseudo-architectural character, its use of sophisticated heraldry and personal badges, the apparent absence of religious iconography on the tombs and, importantly, the collective nature of the programme, key to its interpretation. Using a semiotic approach, a discussion is also offered on the way the various formal, iconographic and conceptual novelties of the princes’ monument impacted on the 15th century monumental landscape in Portugal. Finally, the monument and the chapel housing it are looked at through the prism of the various readings that successive generations of viewers have projected onto it, from the time of its creation to the turn of the 20th century, in order to offer a more comprehensive understanding of the object as it stands today.

Divergent and convergent margins in the north-western Alps - Confrontation to actualistic models

Divergent and convergent margins actualistic models are reviewed and applied to the history of the western Alps. Tethyan rifting history and geometry are analyzed: the northern European margin is considered as an upper plate whereas the southern Apulian margin is a lower plate; the Breche basin is regarded as the former break-away trough; the internal Brianconnais domain represents the northern rift shoulder whilst the more external domains are regarded as the infill of a complex rim basin locally affected by important extension (Valaisan and Vocontain trough). The Schistes lustres and ophiolites of the Tsate nappe are compared to an accretionary prism: the imbrication of this nappe elements is regarded as a direct consequence of the accretionary phenomena already active in early Cretaceous; the Gets/Simme complex could orginate from a more internal part of the accretionary prism. Some eclogitic basements represent the former Apulian margin substratum (Sesia) others (Mont-Rose) are interpreted as the former edge of the European margin. The history of the closing Tethyan domain is analyzed and the remaining problems concerning the cinematics, the presence/absence of a volcanic arc and the eoalpine metamorphism are discussed.

Pulmonary hypertension in Switzerland: treatment and clinical course.

The prognosis of pulmonary hypertension (PH), especially idiopathic pulmonary arterial hypertension (IPAH), has improved during the recent years. The Swiss Registry for PH represents the collaboration of the various centres in Switzerland dealing with PH and serves as an important tool in quality control. The objective of the study was to describe the treatment and clinical course of this orphan disease in Switzerland. We analyzed data from 222 of 252 adult patients, who were included in the registry between January 1999 and December 2004 and suffered from either PAH, PH associated with lung diseases or chronic thromboembolic PH (CTEPH) with respect to the following data: NYHA class, six-minute walking distance (6-MWD), haemodynamics, treatments and survival. If compared with the calculated expected figures the one, two and three year mean survivals in IPAH increased from 67% to 89%, from 55% to 78% and from 46% to 73%, respectively. Most patients (90%) were on oral or inhaled therapy and only 10 patients necessitated lung transplantation. Even though pulmonary endarterectomy (PEA) was performed in only 7 patients during this time, the survival in our CTEPH cohort improved compared with literature data and seems to approach outcomes usually seen after PEA. The 6-MWD increased maximally by 52 m and 59 m in IPAH and CTEPH, respectively, but in the long term returned to or below baseline values, despite the increasing use of multiple specific drugs (overall in 51% of IPAH and 29% of CTEPH). Our national registry data indicate that the overall survival of IPAH and presumably CTEPH seems to have improved in Switzerland. Although the 6-MWD improved transiently, it decreased in the long term despite specific and increasingly combined drug treatment. Our findings herewith underscore the progressive nature of the diseases and the need for further intense research in the field.

Epidemiology of pulmonary hypertension: new data from the Swiss registry.

since 1999 data from pulmonary hypertension (PH) patients from all PH centres in Switzerland were prospectively collected. We analyse the epidemiological aspects of these data. PH was defined as a mean pulmonary artery pressure of >25 mm Hg at rest or >30 mm Hg during exercise. Patients with pulmonary arterial hypertension (PAH), PH associated with lung diseases, PH due to chronic thrombotic and/or embolic disease (CTEPH), or PH due to miscellaneous disorders were registered. Data from adult patients included between January 1999 and December 2004 were analysed. 250 patients were registered (age 58 +/- 16 years, 104 (41%) males). 152 patients (61%) had PAH, 73 (29%) had CTEPH and 18 (7%) had PH associated with lung disease. Patients <50 years (32%) were more likely to have PAH than patients >50 years (76% vs. 53%, p <0.005). Twenty-four patients (10%) were lost to followup, 58 patients (26%) died and 150 (66%) survived without transplantation or thrombendarterectomy. Survivors differed from patients who died in the baseline six-minute walking distance (400 m [300-459] vs. 273 m [174-415]), the functional impairment (NYHA class III/IV 86% vs. 98%), mixed venous saturation (63% [57-68] vs. 56% [50-61]) and right atrial pressure (7 mm Hg [4-11] vs. 11 mm Hg [4-18]). PH is a disease affecting adults of all ages. The management of these patients in specialised centres guarantees a high quality of care. Analysis of the registry data could be an instrument for quality control and might help identify weak points in assessment and treatment of these patients.

Geology of the Gander Group, Gander River Ultramafic Belt and Davidsville Group in the Jonathan's Pond - Weir's Pond Area, northeast Newfoundland /

The study area is situated in NE Newfoundland between Gander Lake and the north coast and on the boundary between the Gander and Botwood tectonostratigraphic zones (Williams et al., 1974). The area is underlain by three NE trending units; the Gander Group, the Gander River Ultramafic Belt (the GRUB) and the Davidsville Group. The easternmost Gander Group consists of a thick, psammitic unit composed predominantly of psammitic schist and a thinner, mixed unit of semipelitic and pelitic schist with minor psammite. The mixed unit may stratigraphically overlie the psammitic unit or be a lateral facies equivalent of the latter. No fossils have been recovered from the Gander Group. The GRUB is a terrain of mafic and ultramafic plutonic rocks with minor pillow lava and plagiogranite. It is interpreted to be a dismembered ophiolite in thrust contact with the Gander Group. The westernmost Davidsville Group consists of a basal conglomerate, believed deposited unconformably upon the GRUB from which it was derived, and an upper unit of greywacke and slate, mostly of turbidite origin, with minor limestone and calcareous sandstone. The limestone, which lies near the base of the unit, contains Upper Llanvirn to Lower Llandeilo fossils. The Gander and Davidsville Groups display distinctly different sedimentological , structural and metamorphic histories. The Gander Group consists of quartz-rich, relatively mature sediment. It has suffered three pre-Llanvirn deformations, of which the main deformation, Dp produced a major, NE-N-facing recumbent anticline in the southern part of the study area. Middle greenschist conditions existed from D^ to D- with growth of metamorphic minerals during each dynamic and static phase. In contrast, the mineralogically immature Davidsville Group sediment contains abundant mafic and ultramafic detritus which is absent from the Gander Group. The Davidsville Group displays the effects of a single penetrative deformation with localized D_ and D_ features, all of which can be shown to postdate D_ in the Gander Group. Rotation of the flat Gander S- into a subvertical orientation near the contact with the GRUB and the Davidsville Group is believed to be a Davidsville D^ feature. Regional metamorphism in the Davidsville Group is lower greenschist with a single growth phase, MS . These sedimentological, structural and metamorphic differences between the Gander and Davidsville Groups persist even where the GRUB is absent and the two units are in contact, indicating that the tectonic histories of the Gander and Davidsville Groups are distinctly different. Structural features in the GRUB, locally the result of multiple deformations, may be the result of Gander and/or Davidsville deformations. Metamorphism is in the greenschist facies. Geochemical analyses of the pillow lava suggest that these rocks were formed in a back-arc basin. Mafic intrusives in the Gander Group appear to be the result of magraatism separate from that producing the pillow lava. The Gander Group is interpreted to be a continental rise prism deposited on the eastern margin of the Late Precambrian-Lower Paleozoic lapetus Ocean. The GRUB, oceanic crust possibly formed in a marginal basin to the west, is believed to have been thrust eastward over the Gander Group, deforming the latter, during the pre-Llanvirnian, possibly Precambrian, Ganderian Orogeny. The Middle Ordovician and younger Davidsville Group was derived from, and deposited unconformably on, this deformed terrain. Deformation of the Davidsville Group occurred during the Middle Devonian Acadian Orogeny.

Sedimentology and stratigraphy of the Lower Silurian Grimsby formation, in subsurface, Lake Erie and Southwestern Ontario

Since the first offshore Lake Erie well was drilled in 1941, the Grimsby and Thorold formations of the Cataract Group have been economically important to the oil and gas industry of Ontario. The Cataract Group provides a significant amount of Ontario's gas production primarily from wells located on Lake Erie. The Grimsby - Thorold formations are the result of nearshore estuarine processes influenced by tides on a prograding shelf and are composed of subtidal channel complexes, discrete tidal channels, mud flats and non-marine deposits. Deposition was related to a regressive - transgressive cycle associated with eustatic sea level changes caused by the melting and resurgence of continental glaciation centred in Africa in the Late Ordovician/Early Silurian. Grimsby deposition began during a regression with the deposition of subtidal channel complexes incised into the marine deposits of the Cabot Head Formation. The presence of mud drapes and mud couplets suggest that these deposits were influenced by tides. These deposits dominate the lower half of the Grimsby. Deposition continued with a change from these subtidal channel complexes to laterally migrating, discrete, shallow tidal channels and mud flats. These were in turn overlain by the non-marine deposits of the Thorold Formation. Grimsby - Thorold deposition ended with a major transgression replacing siliciclastic deposition with primarily carbonate deposition. Sediment was sourced from the east and southeast and associated with a continuation of the Taconic Orogeny into the Early Silurian. The fluvial head of the estuary prograded from a shoreline that was located in western New York and western Pennsylvania running NNE-SSW and then turning NW-SE and paralleling the present day Lake Erie shoreline. iii The facies attributed to the Grimsby - Thorold formations can be ascribed to the three zones within the tripartite zonation suggested by Dalrymple et ale (1992) for estuaries, that is, a marine-dominated facies, a mixed energy facies, and a facies that is dominated by fluvial processes. Also, sediments within the Grimsby - Thorold are commonly fining upwards sequences which are common in estuarine settings whereas deltaic deposits are normally composed of coarsening upwards sequences in a vertical wedge shape with coarser material near the head. The only coarsening observed was in the Thorold Formation and attributed to non-marine deposition by palynological evidence. The presence of a lag deposit at the base of the sediments of the Grimsby Thorold formations suggests that they were incised into the Cabot Head Formation. Further, the thickness of Early Silurian sediments located between the top of the Queenston Formation, where Early Silurian sedimentation began, to the top of the Reynales - Irondequoit formation are constant whether the Grimsby - Thorold formations are present or not. Also, cross-sections using a sand body located in the Cabot Head Formation for correlation further imply that the Grimsby Formation has been incised into the previous deposits of the Cabot Head.

Elucidation of a mechanism of cell lysis by chorhexidine : a biophysical approach

Chlorhexidine is an effective antiseptic used widely in disinfecting products (hand soap), oral products (mouthwash), and is known to have potential applications in the textile industry. Chlorhexidine has been studied extensively through a biological and biochemical lens, showing evidence that it attacks the semipermeable membrane in bacterial cells. Although extremely lethal to bacterial cells, the present understanding of the exact mode of action of chlorhexidine is incomplete. A biophysical approach has been taken to investigate the potential location of chlorhexidine in the lipid bilayer. Deuterium nuclear magnetic resonance was used to characterize the molecular arrangement of mixed phospholipid/drug formulations. Powder spectra were analyzed using the de-Pake-ing technique, a method capable of extracting both the orientation distribution and the anisotropy distribution functions simultaneously. The results from samples of protonated phospholipids mixed with deuterium-labelled chlorhexidine are compared to those from samples of deuterated phospholipids and protonated chlorhexidine to determine its location in the lipid bilayer. A series of neutron scattering experiments were also conducted to study the biophysical interaction of chlorhexidine with a model phospholipid membrane of DMPC, a common saturated lipid found in bacterial cell membranes. The results found the hexamethylene linker to be located at the depth of the glycerol/phosphate region of the lipid bilayer. As drug concentration was increased in samples, a dramatic decrease in bilayer thickness was observed. Differential scanning calorimetry experiments have revealed a depression of the DMPC bilayer gel-to-lamellar phase transition temperature with an increasing drug concentration. The enthalpy of the transition remained the same for all drug concentrations, indicating a strictly drug/headgroup interaction, thus supporting the proposed location of chlorhexidine. In combination, these results lead to the hypothesis that the drug is folded approximately in half on its hexamethylene linker, with the hydrophobic linker at the depth of the glycerol/phosphate region of the lipid bilayer and the hydrophilic chlorophenyl groups located at the lipid headgroup. This arrangement seems to suggest that the drug molecule acts as a wedge to disrupt the bilayer. In vivo, this should make the cell membrane leaky, which is in agreement with a wide range of bacteriological observations.

Micromorphology of Last Glacial Maximum Grounding line sediments around the Antarctic Peninsula

Sediments recovered from seven Last Glacial Maximum grounding lines sites, around the Antarctic Peninsula, were analyzed using micromorphology. This is the first evidence that grounding line sediments from around the Antarctic Peninsula have complex deformational histories and subglacial origins. It was determined that grounding zone wedge contain multiple units, or diamicton layers, with homogenized boundaries. The multiple diamicton units / layers are due to the accretionary formation of a grounding line wedge. All the sediments were deposited via deformation, and continual reincorporation, homogenization of lower diamicton layers by upper diamicton layers produced what macroscopically appeared to be a single massive diamicton unit. The morainal ridge that was sampled, alternatively, is composed of a single unit, or diamicton layer, that was subglacial in origin and believed to have been pushed out to form a ridge that was subsequently deformed via glacial push.

Internet et le paradigme juridictionnel

"Mémoire présenté à la faculté des études supérieures en vue de l'obtention du grade de maîtrise en droit (LL.M.)"

Le motif d’empaquetage le long du sillon: une nouvelle entité structurale récurrente dans les ARN ribosomiques

La plupart des molécules d’ARN doivent se replier en structure tertiaire complexe afin d’accomplir leurs fonctions biologiques. Cependant, les déterminants d’une chaîne de polynucléotides qui sont nécessaires à son repliement et à ses interactions avec d’autres éléments sont essentiellement inconnus. L’établissement des relations structure-fonction dans les grandes molécules d’ARN passe inévitablement par l’analyse de chaque élément de leur structure de façon individuelle et en contexte avec d’autres éléments. À l’image d’une construction d’immeuble, une structure d’ARN est composée d’unités répétitives assemblées de façon spécifique. Les motifs récurrents d’ARN sont des arrangements de nucléotides retrouvés à différents endroits d’une structure tertiaire et possèdent des conformations identiques ou très similaires. Ainsi, une des étapes nécessaires à la compréhension de la structure et de la fonction des molécules d’ARN consiste à identifier de façon systématique les motifs récurrents et d’en effectuer une analyse comparative afin d’établir la séquence consensus. L’analyse de tous les cas d’empaquetage de doubles hélices dans la structure du ribosome a permis l’identification d’un nouvel arrangement nommé motif d’empaquetage le long du sillon (AGPM) (along-groove packing motif). Ce motif est retrouvé à 14 endroits dans la structure du ribosome de même qu’entre l’ARN ribosomique 23S et les molécules d’ARN de transfert liées aux sites ribosomaux P et E. Le motif se forme par l’empaquetage de deux doubles hélices via leur sillon mineur. Le squelette sucre-phosphate d’une hélice voyage le long du sillon mineur de l’autre hélice et vice versa. Dans chacune des hélices, la région de contact comprend quatre paires de bases. L’empaquetage le plus serré est retrouvé au centre de l’arrangement où l’on retrouve souvent une paire de bases GU dans une hélice interagissant avec une paire de bases Watson-Crick (WC) dans l’autre hélice. Même si la présence des paires de bases centrales GU versus WC au centre du motif augmente sa stabilité, d’autres alternatives existent pour différents représentants du motif. L’analyse comparative de trois librairies combinatoires de gènes d’AGPM, où les paires de bases centrales ont été variées de manière complètement aléatoire, a montré que le contexte structural influence l’étendue de la variabilité des séquences de nucléotides formant les paires de bases centrales. Le fait que l’identité des paires de bases centrales puisse varier suggérait la présence d’autres déterminants responsables au maintien de l’intégrité du motif. L’analyse de tous les contacts entre les hélices a révélé qu’en dehors du centre du motif, les interactions entre les squelettes sucre-phosphate s’effectuent via trois contacts ribose-ribose. Pour chacun de ces contacts, les riboses des nucléotides qui interagissent ensemble doivent adopter des positions particulières afin d’éviter qu’ils entrent en collision. Nous montrons que la position de ces riboses est modulée par des conformations spécifiques des paires de bases auxquelles ils appartiennent. Finalement, un autre motif récurrent identifié à l’intérieur même de la structure de trois cas d’AGPM a été nommé « adenosine-wedge ». Son analyse a révélé que ce dernier est lui-même composé d’un autre arrangement, nommé motif triangle-NAG (NAG-triangle). Nous montrons que le motif « adenosine-wedge » représente un arrangement complexe d’ARN composé de quatre éléments répétitifs, c’est-à-dire des motifs AGPM, « hook-turn », « A-minor » et triangle-NAG. Ceci illustre clairement l’arrangement hiérarchique des structures d’ARN qui peut aussi être observé pour d’autres motifs d’ARN. D’un point de vue plus global, mes résultats enrichissent notre compréhension générale du rôle des différents types d’interactions tertiaires dans la formation des molécules d’ARN complexes.

Argentina’s Worker-Recovered Factories : strategies and survival

Les usines récupérées par les ouvriers en Argentine sont devenues un mouvement social emblématique symbolisant l'un des aspects de la révolte sociale entourant la crise économique de 2001-2002. Les usines récupérées sont des entreprises abandonnées par leurs propriétaires originaux ou déclarées faillite, laissant derrières elles des salaires et des dettes impayés. Par conséquence, les ouvriers ont commencé à récupérer leurs usines; reprenant la production sans leurs anciens patrons, sous, et au profit de la gestion collective des ouvriers. Le mouvement est remarquable pour sa rémunération égalitaire et sa gestion horizontale. Ce travail examine la continuité des usines récupérées et ceci à travers l'évolution sociale, politique et économique du paysage de l'Argentine. Il évalue également l'impact du mouvement en tant que défi aux modes économiques de production hégémoniques et orientés vers le marché. En supposant que l'avenir du mouvement dépend de deux ensembles de facteurs, le rapport analyse les facteurs internes à travers le prisme de la théorie de mobilisation des ressources, ainsi que les facteurs externes à travers la perspective de la théorie de la structure de l'opportunité politique. Le travail conclut que la situation actuelle se trouve dans une impasse dans laquelle le mouvement a gagné l'acceptation institutionnelle, mais a échoué d'effectuer le changement structurel favorisant ses pratiques et garantissant la sécurité à long terme. Il argumente que le mouvement doit consolider certains aspects combatifs. Il doit consolider sa nouvelle identité en tant que mouvement social et forger des alliances stratégiques et tactiques tout en préservant son autonomie.