Differential expression and modification of neurofilament triplet proteins during cat cerebellar development.

Neurofilament (NF) proteins consist of three subunits of different molecular weights defined as NF-H, NF-M, and NF-L. They are typical structures of the neuronal cytoskeleton. Their immunocytochemical distribution during postnatal development of cat cerebellum was studied with several monoclonal and polyclonal antibodies against phosphorylated or unmodified sites. Expression and distribution of the triplet neurofilament proteins changed with maturation. Afferent mossy and climbing fibers in the medullary layer contained NF-M and NF-L already at birth, whereas NF-H appeared later. Within the first three postnatal weeks, all three subunits appeared in mossy and climbing fibers in the internal granular and molecular layers and in the axons of Purkinje cells. Axons of local circuit neurons such as basket cells expressed these proteins at the end of the first month, whereas parallel fibers expressed them last, at the beginning of the third postnatal month. Differential localization was especially observed for NF-H. Depending on phosphorylation, NF-H proteins were found in different axon types in climbing, mossy, and basket fibers or additionally in parallel fibers. A nonphosphorylated NF-H subunit was exclusively located in some Purkinje cells at early developmental stages and in some smaller interneurons later. A novel finding is the presence of a phosphorylation site in the NF-H subunit that is localized in dendrites of Purkinje cells but not in axons. Expression and phosphorylation of the NF-H subunit, especially, is cell-type specific and possibly involved in the adult-type stabilization of the axonal and dendritic cytoskeleton.

Development and use of a high-fidelity simulator for fetal endotracheal balloon occlusion (FETO) insertion and removal.

Objectives The objective of this article is to describe the development of an anatomically accurate simulator in order to aid the training of a perinatal team in the insertion and removal of a fetal endoscopic tracheal occlusion (FETO) balloon in the management of prenatally diagnosed congenital diaphragmatic hernia. Methods An experienced perinatal team collaborated with a medical sculptor to design a fetal model for the FETO procedure. Measurements derived from 28-week fetal magnetic resonance imaging were used in the development of an anatomically precise simulated airway within a silicone rubber preterm fetal model. Clinician feedback was then used to guide multiple iterations of the model with serial improvements in the anatomic accuracy of the simulator airway. Results An appropriately sized preterm fetal mannequin with a high-fidelity airway was developed. The team used this model to develop surgical skills with balloon insertion, and removal, and to prepare the team for an integrated response to unanticipated delivery with the FETO balloon still in situ. Conclusions This fetal mannequin aided in the ability of a fetal therapy unit to offer the FETO procedure at their center for the first time. This model may be of benefit to other perinatal centers planning to offer this procedure.

Development of a system for 3D high-resolution seismic reflection profiling on lakes

A high-resolution three-dimensional (3D) seismic reflection system for small-scale targets in lacustrine settings has been developed. Its main characteristics include navigation and shot-triggering software that fires the seismic source at regular distance intervals (max. error of 0.25 m) with real-time control on navigation using differential GPS (Global Positioning System). Receiver positions are accurately calculated (error < 0.20 m) with the aid of GPS antennas attached to the end of each of three 24-channel streamers. Two telescopic booms hold the streamers at a distance of 7.5 m from each other. With a receiver spacing of 2.5 m, the bin dimension is 1.25 m in inline and 3.75 m in crossline direction. To test the system, we conducted a 3D survey of about 1 km(2) in Lake Geneva, Switzerland, over a complex fault zone. A 5-m shot spacing resulted in a nominal fold of 6. A double-chamber bubble-cancelling 15/15 in(3) air gun (40-650 Hz) operated at 80 bars and 1 m depth gave a signal penetration of 300 m below water bottom and a best vertical resolution of 1.1 m. Processing followed a conventional scheme, but had to be adapted to the high sampling rates, and our unconventional navigation data needed conversion to industry standards. The high-quality data enabled us to construct maps of seismic horizons and fault surfaces in three dimensions. The system proves to be well adapted to investigate complex structures by providing non-aliased images of reflectors with dips up to 30 degrees.

The immune space: a concept and template for rationalizing vaccine development.

Abstract Empirical testing of candidate vaccines has led to the successful development of a number of lifesaving vaccines. The advent of new tools to manipulate antigens and new methods and vectors for vaccine delivery has led to a veritable explosion of potential vaccine designs. As a result, selection of candidate vaccines suitable for large-scale efficacy testing has become more challenging. This is especially true for diseases such as dengue, HIV, and tuberculosis where there is no validated animal model or correlate of immune protection. Establishing guidelines for the selection of vaccine candidates for advanced testing has become a necessity. A number of factors could be considered in making these decisions, including, for example, safety in animal and human studies, immune profile, protection in animal studies, production processes with product quality and stability, availability of resources, and estimated cost of goods. The "immune space template" proposed here provides a standardized approach by which the quality, level, and durability of immune responses elicited in early human trials by a candidate vaccine can be described. The immune response profile will demonstrate if and how the candidate is unique relative to other candidates, especially those that have preceded it into efficacy testing and, thus, what new information concerning potential immune correlates could be learned from an efficacy trial. A thorough characterization of immune responses should also provide insight into a developer's rationale for the vaccine's proposed mechanism of action. HIV vaccine researchers plan to include this general approach in up-selecting candidates for the next large efficacy trial. This "immune space" approach may also be applicable to other vaccine development endeavors where correlates of vaccine-induced immune protection remain unknown.

FRAX(®) Bone Mineral Density Task Force of the 2010 Joint International Society for Clinical Densitometry International Osteoporosis Foundation Position Development Conference.

FRAX(®) is a fracture risk assessment algorithm developed by the World Health Organization in cooperation with other medical organizations and societies. Using easily available clinical information and femoral neck bone mineral density (BMD) measured by dual-energy X-ray absorptiometry (DXA), when available, FRAX(®) is used to predict the 10-year probability of hip fracture and major osteoporotic fracture. These values may be included in country specific guidelines to aid clinicians in determining when fracture risk is sufficiently high that the patient is likely to benefit from pharmacological therapy to reduce that risk. Since the introduction of FRAX(®) into clinical practice, many practical clinical questions have arisen regarding its use. To address such questions, the International Society for Clinical Densitometry (ISCD) and International Osteoporosis Foundations (IOF) assigned task forces to review the best available medical evidence and make recommendations for optimal use of FRAX(®) in clinical practice. Questions were identified and divided into three general categories. A task force was assigned to investigating the medical evidence in each category and developing clinically useful recommendations. The BMD Task Force addressed issues that included the potential use of skeletal sites other than the femoral neck, the use of technologies other than DXA, and the deletion or addition of clinical data for FRAX(®) input. The evidence and recommendations were presented to a panel of experts at the ISCD-IOF FRAX(®) Position Development Conference, resulting in the development of ISCD-IOF Official Positions addressing FRAX(®)-related issues.

Development of a 3D very high-resolution seismic reflection system for lacustrine settings - a case study over a thrust fault zone in Lake Geneva

Un système efficace de sismique tridimensionnelle (3-D) haute-résolution adapté à des cibles lacustres de petite échelle a été développé. Dans le Lac Léman, près de la ville de Lausanne, en Suisse, des investigations récentes en deux dimension (2-D) ont mis en évidence une zone de faille complexe qui a été choisie pour tester notre système. Les structures observées incluent une couche mince (<40 m) de sédiments quaternaires sub-horizontaux, discordants sur des couches tertiaires de molasse pentées vers le sud-est. On observe aussi la zone de faille de « La Paudèze » qui sépare les unités de la Molasse du Plateau de la Molasse Subalpine. Deux campagnes 3-D complètes, d?environ d?un kilomètre carré, ont été réalisées sur ce site de test. La campagne pilote (campagne I), effectuée en 1999 pendant 8 jours, a couvert 80 profils en utilisant une seule flûte. Pendant la campagne II (9 jours en 2001), le nouveau système trois-flûtes, bien paramétrés pour notre objectif, a permis l?acquisition de données de très haute qualité sur 180 lignes CMP. Les améliorations principales incluent un système de navigation et de déclenchement de tirs grâce à un nouveau logiciel. Celui-ci comprend un contrôle qualité de la navigation du bateau en temps réel utilisant un GPS différentiel (dGPS) à bord et une station de référence près du bord du lac. De cette façon, les tirs peuvent être déclenchés tous les 5 mètres avec une erreur maximale non-cumulative de 25 centimètres. Tandis que pour la campagne I la position des récepteurs de la flûte 48-traces a dû être déduite à partir des positions du bateau, pour la campagne II elle ont pu être calculées précisément (erreur <20 cm) grâce aux trois antennes dGPS supplémentaires placées sur des flotteurs attachés à l?extrémité de chaque flûte 24-traces. Il est maintenant possible de déterminer la dérive éventuelle de l?extrémité des flûtes (75 m) causée par des courants latéraux ou de petites variations de trajet du bateau. De plus, la construction de deux bras télescopiques maintenant les trois flûtes à une distance de 7.5 m les uns des autres, qui est la même distance que celle entre les lignes naviguées de la campagne II. En combinaison avec un espacement de récepteurs de 2.5 m, la dimension de chaque «bin» de données 3-D de la campagne II est de 1.25 m en ligne et 3.75 m latéralement. L?espacement plus grand en direction « in-line » par rapport à la direction «cross-line» est justifié par l?orientation structurale de la zone de faille perpendiculaire à la direction «in-line». L?incertitude sur la navigation et le positionnement pendant la campagne I et le «binning» imprécis qui en résulte, se retrouve dans les données sous forme d?une certaine discontinuité des réflecteurs. L?utilisation d?un canon à air à doublechambre (qui permet d?atténuer l?effet bulle) a pu réduire l?aliasing observé dans les sections migrées en 3-D. Celui-ci était dû à la combinaison du contenu relativement haute fréquence (<2000 Hz) du canon à eau (utilisé à 140 bars et à 0.3 m de profondeur) et d?un pas d?échantillonnage latéral insuffisant. Le Mini G.I 15/15 a été utilisé à 80 bars et à 1 m de profondeur, est mieux adapté à la complexité de la cible, une zone faillée ayant des réflecteurs pentés jusqu?à 30°. Bien que ses fréquences ne dépassent pas les 650 Hz, cette source combine une pénétration du signal non-aliasé jusqu?à 300 m dans le sol (par rapport au 145 m pour le canon à eau) pour une résolution verticale maximale de 1.1 m. Tandis que la campagne I a été acquise par groupes de plusieurs lignes de directions alternées, l?optimisation du temps d?acquisition du nouveau système à trois flûtes permet l?acquisition en géométrie parallèle, ce qui est préférable lorsqu?on utilise une configuration asymétrique (une source et un dispositif de récepteurs). Si on ne procède pas ainsi, les stacks sont différents selon la direction. Toutefois, la configuration de flûtes, plus courtes que pour la compagne I, a réduit la couverture nominale, la ramenant de 12 à 6. Une séquence classique de traitement 3-D a été adaptée à l?échantillonnage à haute fréquence et elle a été complétée par deux programmes qui transforment le format non-conventionnel de nos données de navigation en un format standard de l?industrie. Dans l?ordre, le traitement comprend l?incorporation de la géométrie, suivi de l?édition des traces, de l?harmonisation des «bins» (pour compenser l?inhomogénéité de la couverture due à la dérive du bateau et de la flûte), de la correction de la divergence sphérique, du filtrage passe-bande, de l?analyse de vitesse, de la correction DMO en 3-D, du stack et enfin de la migration 3-D en temps. D?analyses de vitesse détaillées ont été effectuées sur les données de couverture 12, une ligne sur deux et tous les 50 CMP, soit un nombre total de 600 spectres de semblance. Selon cette analyse, les vitesses d?intervalles varient de 1450-1650 m/s dans les sédiments non-consolidés et de 1650-3000 m/s dans les sédiments consolidés. Le fait que l?on puisse interpréter plusieurs horizons et surfaces de faille dans le cube, montre le potentiel de cette technique pour une interprétation tectonique et géologique à petite échelle en trois dimensions. On distingue cinq faciès sismiques principaux et leurs géométries 3-D détaillées sur des sections verticales et horizontales: les sédiments lacustres (Holocène), les sédiments glacio-lacustres (Pléistocène), la Molasse du Plateau, la Molasse Subalpine de la zone de faille (chevauchement) et la Molasse Subalpine au sud de cette zone. Les couches de la Molasse du Plateau et de la Molasse Subalpine ont respectivement un pendage de ~8° et ~20°. La zone de faille comprend de nombreuses structures très déformées de pendage d?environ 30°. Des tests préliminaires avec un algorithme de migration 3-D en profondeur avant sommation et à amplitudes préservées démontrent que la qualité excellente des données de la campagne II permet l?application de telles techniques à des campagnes haute-résolution. La méthode de sismique marine 3-D était utilisée jusqu?à présent quasi-exclusivement par l?industrie pétrolière. Son adaptation à une échelle plus petite géographiquement mais aussi financièrement a ouvert la voie d?appliquer cette technique à des objectifs d?environnement et du génie civil.<br/><br/>An efficient high-resolution three-dimensional (3-D) seismic reflection system for small-scale targets in lacustrine settings was developed. In Lake Geneva, near the city of Lausanne, Switzerland, past high-resolution two-dimensional (2-D) investigations revealed a complex fault zone (the Paudèze thrust zone), which was subsequently chosen for testing our system. Observed structures include a thin (<40 m) layer of subhorizontal Quaternary sediments that unconformably overlie southeast-dipping Tertiary Molasse beds and the Paudèze thrust zone, which separates Plateau and Subalpine Molasse units. Two complete 3-D surveys have been conducted over this same test site, covering an area of about 1 km2. In 1999, a pilot survey (Survey I), comprising 80 profiles, was carried out in 8 days with a single-streamer configuration. In 2001, a second survey (Survey II) used a newly developed three-streamer system with optimized design parameters, which provided an exceptionally high-quality data set of 180 common midpoint (CMP) lines in 9 days. The main improvements include a navigation and shot-triggering system with in-house navigation software that automatically fires the gun in combination with real-time control on navigation quality using differential GPS (dGPS) onboard and a reference base near the lake shore. Shots were triggered at 5-m intervals with a maximum non-cumulative error of 25 cm. Whereas the single 48-channel streamer system of Survey I requires extrapolation of receiver positions from the boat position, for Survey II they could be accurately calculated (error <20 cm) with the aid of three additional dGPS antennas mounted on rafts attached to the end of each of the 24- channel streamers. Towed at a distance of 75 m behind the vessel, they allow the determination of feathering due to cross-line currents or small course variations. Furthermore, two retractable booms hold the three streamers at a distance of 7.5 m from each other, which is the same distance as the sail line interval for Survey I. With a receiver spacing of 2.5 m, the bin dimension of the 3-D data of Survey II is 1.25 m in in-line direction and 3.75 m in cross-line direction. The greater cross-line versus in-line spacing is justified by the known structural trend of the fault zone perpendicular to the in-line direction. The data from Survey I showed some reflection discontinuity as a result of insufficiently accurate navigation and positioning and subsequent binning errors. Observed aliasing in the 3-D migration was due to insufficient lateral sampling combined with the relatively high frequency (<2000 Hz) content of the water gun source (operated at 140 bars and 0.3 m depth). These results motivated the use of a double-chamber bubble-canceling air gun for Survey II. A 15 / 15 Mini G.I air gun operated at 80 bars and 1 m depth, proved to be better adapted for imaging the complexly faulted target area, which has reflectors dipping up to 30°. Although its frequencies do not exceed 650 Hz, this air gun combines a penetration of non-aliased signal to depths of 300 m below the water bottom (versus 145 m for the water gun) with a maximum vertical resolution of 1.1 m. While Survey I was shot in patches of alternating directions, the optimized surveying time of the new threestreamer system allowed acquisition in parallel geometry, which is preferable when using an asymmetric configuration (single source and receiver array). Otherwise, resulting stacks are different for the opposite directions. However, the shorter streamer configuration of Survey II reduced the nominal fold from 12 to 6. A 3-D conventional processing flow was adapted to the high sampling rates and was complemented by two computer programs that format the unconventional navigation data to industry standards. Processing included trace editing, geometry assignment, bin harmonization (to compensate for uneven fold due to boat/streamer drift), spherical divergence correction, bandpass filtering, velocity analysis, 3-D DMO correction, stack and 3-D time migration. A detailed semblance velocity analysis was performed on the 12-fold data set for every second in-line and every 50th CMP, i.e. on a total of 600 spectra. According to this velocity analysis, interval velocities range from 1450-1650 m/s for the unconsolidated sediments and from 1650-3000 m/s for the consolidated sediments. Delineation of several horizons and fault surfaces reveal the potential for small-scale geologic and tectonic interpretation in three dimensions. Five major seismic facies and their detailed 3-D geometries can be distinguished in vertical and horizontal sections: lacustrine sediments (Holocene) , glaciolacustrine sediments (Pleistocene), Plateau Molasse, Subalpine Molasse and its thrust fault zone. Dips of beds within Plateau and Subalpine Molasse are ~8° and ~20°, respectively. Within the fault zone, many highly deformed structures with dips around 30° are visible. Preliminary tests with 3-D preserved-amplitude prestack depth migration demonstrate that the excellent data quality of Survey II allows application of such sophisticated techniques even to high-resolution seismic surveys. In general, the adaptation of the 3-D marine seismic reflection method, which to date has almost exclusively been used by the oil exploration industry, to a smaller geographical as well as financial scale has helped pave the way for applying this technique to environmental and engineering purposes.<br/><br/>La sismique réflexion est une méthode d?investigation du sous-sol avec un très grand pouvoir de résolution. Elle consiste à envoyer des vibrations dans le sol et à recueillir les ondes qui se réfléchissent sur les discontinuités géologiques à différentes profondeurs et remontent ensuite à la surface où elles sont enregistrées. Les signaux ainsi recueillis donnent non seulement des informations sur la nature des couches en présence et leur géométrie, mais ils permettent aussi de faire une interprétation géologique du sous-sol. Par exemple, dans le cas de roches sédimentaires, les profils de sismique réflexion permettent de déterminer leur mode de dépôt, leurs éventuelles déformations ou cassures et donc leur histoire tectonique. La sismique réflexion est la méthode principale de l?exploration pétrolière. Pendant longtemps on a réalisé des profils de sismique réflexion le long de profils qui fournissent une image du sous-sol en deux dimensions. Les images ainsi obtenues ne sont que partiellement exactes, puisqu?elles ne tiennent pas compte de l?aspect tridimensionnel des structures géologiques. Depuis quelques dizaines d?années, la sismique en trois dimensions (3-D) a apporté un souffle nouveau à l?étude du sous-sol. Si elle est aujourd?hui parfaitement maîtrisée pour l?imagerie des grandes structures géologiques tant dans le domaine terrestre que le domaine océanique, son adaptation à l?échelle lacustre ou fluviale n?a encore fait l?objet que de rares études. Ce travail de thèse a consisté à développer un système d?acquisition sismique similaire à celui utilisé pour la prospection pétrolière en mer, mais adapté aux lacs. Il est donc de dimension moindre, de mise en oeuvre plus légère et surtout d?une résolution des images finales beaucoup plus élevée. Alors que l?industrie pétrolière se limite souvent à une résolution de l?ordre de la dizaine de mètres, l?instrument qui a été mis au point dans le cadre de ce travail permet de voir des détails de l?ordre du mètre. Le nouveau système repose sur la possibilité d?enregistrer simultanément les réflexions sismiques sur trois câbles sismiques (ou flûtes) de 24 traces chacun. Pour obtenir des données 3-D, il est essentiel de positionner les instruments sur l?eau (source et récepteurs des ondes sismiques) avec une grande précision. Un logiciel a été spécialement développé pour le contrôle de la navigation et le déclenchement des tirs de la source sismique en utilisant des récepteurs GPS différentiel (dGPS) sur le bateau et à l?extrémité de chaque flûte. Ceci permet de positionner les instruments avec une précision de l?ordre de 20 cm. Pour tester notre système, nous avons choisi une zone sur le Lac Léman, près de la ville de Lausanne, où passe la faille de « La Paudèze » qui sépare les unités de la Molasse du Plateau et de la Molasse Subalpine. Deux campagnes de mesures de sismique 3-D y ont été réalisées sur une zone d?environ 1 km2. Les enregistrements sismiques ont ensuite été traités pour les transformer en images interprétables. Nous avons appliqué une séquence de traitement 3-D spécialement adaptée à nos données, notamment en ce qui concerne le positionnement. Après traitement, les données font apparaître différents faciès sismiques principaux correspondant notamment aux sédiments lacustres (Holocène), aux sédiments glacio-lacustres (Pléistocène), à la Molasse du Plateau, à la Molasse Subalpine de la zone de faille et la Molasse Subalpine au sud de cette zone. La géométrie 3-D détaillée des failles est visible sur les sections sismiques verticales et horizontales. L?excellente qualité des données et l?interprétation de plusieurs horizons et surfaces de faille montrent le potentiel de cette technique pour les investigations à petite échelle en trois dimensions ce qui ouvre des voies à son application dans les domaines de l?environnement et du génie civil.

Contrast : a concept in language and its development in Medieval Spanish

The purpose of this PhD thesis is to investigate a semantic relation present in the connection of sentences (more specifically: propositional units). This relation, which we refer to as contrast, includes the traditional categories of adversatives - predominantly represented by the connector but in English and pero in Modern Spanish - and concessives, prototypically verbalised through although / aunque. The aim is to describe, analyse and - as far as possible - to explain the emergence and evolution of different syntactic schemes marking contrast during the first three centuries of Spanish (also referred to as Castilian) as a literary language, i.e., from the 13th to the 15th century. The starting point of this question is a commonplace in syntax, whereby the semantic and syntactic complexity of clause linkage correlates with the degree of textual elaboration. In historical linguistics, i.e., applied to the phylogeny of a language, it is commonly referred to as the parataxis hypothesis A crucial part of the thesis is dedicated by the definition of contrast as a semantic relation. Although the label contrast has been used in this sense, mainly in functional grammar and text linguistics, mainstream grammaticography and linguistics remain attached to the traditional categories adversatives and concessives. In opposition to this traditional view, we present our own model of contrast, based on a pragma-semantic description proposed for the analysis of adversatives by Oswald Ducrot and subsequently adopted by Ekkehard König for the analysis of concessives. We refine and further develop this model in order for it to accommodate all, not just the prototypical instances of contrast in Spanish, arguing that the relationship between adversatives and concessives is a marked opposition, i.e., that the higher degree of semantic and syntactic integration of concessives restricts some possible readings that the adversatives may have, but that this difference is almost systematically neutralised by contextual factors, thus justifying the assumption of contrast as a comprehensive onomasiological category. This theoretical focus is completed by a state-of-the-question overview attempting to account for all relevant forms in which contrast is expressed in Medieval Spanish, with the aid of lexicographic and grammaticographical sources, and an empirical study investigating the expression of corpus in a corpus study on the textual functions of contrast in nine Medieval Spanish texts: Cantar de Mio Cid, Libro de Alexandre, Milagros de Nuestra Sehora, Estoria de Espana, Primera Partida, Lapidario, Libro de buen amor, Conde Lucanor, and Corbacho. This corpus is analysed using quantitative and qualitative tools, and the study is accompanied by a series of methodological remarks on how to investigate a pragma-semantic category in historical linguistics. The corpus study shows that the parataxis hypothesis fails to prove from a statistical viewpoint, although a qualitative analysis shows that the use of subordination does increase over time in some particular contexts.