Development of a comprehensive on-line multidimensional high performance column liquid chromatography system for protein and peptide mapping with integrated size selective sample fractionation

Aufbau einer kontinuierlichen, mehrdimensionalen Hochleistungs-flüssigchromatographie-Anlage für die Trennung von Proteinen und Peptiden mit integrierter größenselektiver ProbenfraktionierungEs wurde eine mehrdimensionale HPLC-Trennmethode für Proteine und Peptide mit einem Molekulargewicht von <15 kDa entwickelt.Im ersten Schritt werden die Zielanalyte von höhermolekularen sowie nicht ionischen Bestandteilen mit Hilfe von 'Restricted Access Materialien' (RAM) mit Ionenaustauscher-Funktionalität getrennt. Anschließend werden die Proteine auf einer analytischen Ionenaustauscher-Säule sowie auf Reversed-Phase-Säulen getrennt. Zur Vermeidung von Probenverlusten wurde ein kontinuierlich arbeitendes, voll automatisiertes System auf Basis unterschiedlicher Trenngeschwindigkeiten und vier parallelen RP-Säulen aufgebaut.Es werden jeweils zwei RP-Säulen gleichzeitig, jedoch mit zeitlich versetztem Beginn eluiert, um durch flache Gradienten ausreichende Trennleistungen zu erhalten. Während die dritte Säule regeneriert wird, erfolgt das Beladen der vierte Säule durch Anreicherung der Proteine und Peptide am Säulenkopf. Während der Gesamtanalysenzeit von 96 Minuten werden in Intervallen von 4 Minuten Fraktionen aus der 1. Dimension auf die RP-Säulen überführt und innerhalb von 8 Minuten getrennt, wobei 24 RP-Chromatogramme resultieren.Als Testsubstanzen wurden u.a. Standardproteine, Proteine und Peptide aus humanem Hämofiltrat sowie aus Lungenfibroblast-Zellkulturüberständen eingesetzt. Weiterhin wurden Fraktionen gesammelt und mittels MALDI-TOF Massenspektrometrie untersucht. Bei einer Injektion wurden in den 24 RP-Chromatogrammen mehr als 1000 Peaks aufgelöst. Der theoretische Wert der Peakkapazität liegt bei ungefähr 3000.

Hox genes and the regulation of programmed cell death in the embryonic central nervous system of Drosophila melanogaster

This study deals with the function and regulation of programmed cell death, or apoptosis, in the development of the embryonic central nervous system of Drosophila melanogaster. The first part provides a description of apoptosis-deficient embryos, which showed that preventing apoptosis does not cause gross morphological defects in the CNS, as it appears well organized despite the presence of too many cells. An analysis of the incidence and pattern of apoptosis over the course of development discloses a partly very orderly pattern suggesting tight spatio-temporal control, but also reveals random apoptotic cells, which suggests a certain amount of plasticity in the embryo. This analysis also allowed precise identification of some of the dying neural cells in the embryo, and establishment of single cell models for studying regulation of segment-specific apoptosis in the embryonic CNS. In the second part of the work, further investigations into mechanisms controlling segment-specific apoptosis revealed the involvement of two Hox genes, Antennapedia (Antp) and Ultrabithorax (Ubx), in this process. Hox genes control the formation of segment-specific structures in their domains of expression, but also regulate organ and tissue morphogenesis. The study presented here shows that Antp and Ubx play antagonistic roles in motoneuron survival in the embryo. Ubx expression in the CNS is strongly upregulated at a late point in development, when most cells have begun to differentiate. This upregulation shortly precedes Ubx-dependent, segment-specific apoptosis of two differentiated motoneurons. It could further be demonstrated that Antp is required for proper development of the NB7-3 lineage and for survival of the NB7-3 motoneuron in the anterior thoracic segments. In segments where Antp and Ubx expression overlaps, Ubx counteracts the anti-apoptotic function of Antp, resulting in cell death. Thus, these two Hox genes play opposing roles in the survival of differentiated neurons in the late developing nervous system. They thereby contribute to establishment of correct connections between outward-projecting neurons and their targets, which is crucial for the assembly of functional neural circuits, as these have to fulfill region-specific locomotion and sensory requirements along the antero-posterior body axis.

From Radio Channel Modeling to a System Level Perspective in Body-Centric Communications

Body-centric communications are emerging as a new paradigm in the panorama of personal communications. Being concerned with human behaviour, they are suitable for a wide variety of applications. The advances in the miniaturization of portable devices to be placed on or around the body, foster the diffusion of these systems, where the human body is the key element defining communication characteristics. This thesis investigates the human impact on body-centric communications under its distinctive aspects. First of all, the unique propagation environment defined by the body is described through a scenario-based channel modeling approach, according to the communication scenario considered, i.e., on- or on- to off-body. The novelty introduced pertains to the description of radio channel features accounting for multiple sources of variability at the same time. Secondly, the importance of a proper channel characterisation is shown integrating the on-body channel model in a system level simulator, allowing a more realistic comparison of different Physical and Medium Access Control layer solutions. Finally, the structure of a comprehensive simulation framework for system performance evaluation is proposed. It aims at merging in one tool, mobility and social features typical of the human being, together with the propagation aspects, in a scenario where multiple users interact sharing space and resources.

Feeding entrainment of locomotor activity and clock gene expression in Senegalese sole, Solea senegalensis

The present study was conducted to investigate the influence of restricted food access on Solea senegalensis behaviour and daily expression of clock genes in central (diencephalon and optic tectum) and pheripheral (liver) tissues. The Senegalese sole is a marine teleost fish belonging to the Class of Actinopterygii, Order Pleuronectiformes and Family Soleidae. Its geographical distribution in the Mediterranean sea is fairly broad, covering the south and east of the Iberian Peninsula, the North of Africa and Middle East until the coast of Turkey. From a commercial perspective Solea senegalensis has acquired in recent years, a key role in aquacolture industry of the Iberian Peninsula. The Senegalese sole is also acquiring an important relevance in chronobiological studies as the number of published works focused on the sole circadian system has increased in the last few years. The molecular mechanisms underlying sole circadian rhythms has also been explored recently, both in adults and developing sole. Moreover, the consideration of the Pleuronectiformes Order as one of the most evolved teleost groups make the Senegalese sole a species of high interest under a comparative and phylogenetic point of view. All these facts have reinforced the election of Senegalese sole as model species for the present study. The animals were kept under 12L:12D photoperiod conditions and divided into three experimental groups depending on the feeding time: fed at midlight (ML), middark (MD) or random (RND) times. Throughout the experiment, the existence of a daily activity rhythm and it synchronization to the light-dark and feeding cycles was checked. To this end locomotor activity was registred by means of two infrared photocells placed in pvc tube 10 cm below the water surface (upper photocell) and the other one was located 10 cm above the bottom of the tank (bottom photocell). The photocell were connected to a computer so that every time a fish interrupted the infrared light beam, it produced an output signal that was recorded. The number of light beam interruptions was stored every 10 minutes by specialized software for data acquisition.

TCP a ritrasmissione asimmetrica anticipata su Access Point WiFi

La perdita di pacchetti durante una trasmissione su una rete Wireless influisce in maniera fondamentale sulla qualità del collegamento tra due End-System. Lo scopo del progetto è quello di implementare una tecnica di ritrasmissione asimmetrica anticipata dei pacchetti perduti, in modo da minimizzare i tempi di recupero dati e migliorare la qualità della comunicazione. Partendo da uno studio su determinati tipi di ritrasmissione, in particolare quelli implementati dal progetto ABPS, Always Best Packet Switching, si è maturata l'idea che un tipo di ritrasmissione particolarmente utile potrebbe avvenire a livello Access Point: nel caso in cui la perdita di pacchetti avvenga tra l'AP e il nodo mobile che vi è collegato via IEEE802.11, invece che attendere la ritrasmissione TCP e Effettuata dall'End-System sorgente è lo stesso Access Point che e effettua una ritrasmissione verso il nodo mobile per permettere un veloce recupero dei dati perduti. Tale funzionalità stata quindi concettualmente divisa in due parti, la prima si riferisce all'applicazione che si occupa della bufferizzazione di pacchetti che attraversano l'AP e della loro copia in memoria per poi ritrasmetterli in caso di segnalazione di mancata acquisizione, la seconda riguardante la modifica al kernel che permette la segnalazione anticipata dell'errore. E' già stata sviluppata un'applicazione che prevede una ritrasmissione anticipata da parte dell'Access Point Wifi, cioè una ritrasmissione prima che la notifica di avvenuta perdita raggiunga l'end-point sorgente e appoggiata su un meccanismo di simulazione di Error Detection. Inoltre è stata anche realizzata la ritrasmissione asincrona e anticipata del TCP. Questo documento tratta della realizzazione di una nuova applicazione che fornisca una più effciente versione del buffer di pacchetti e utilizzi il meccanismo di una ritrasmissione asimmetrica e anticipata del TCP, cioè attivare la ritrasmissione su richiesta del TCP tramite notifiche di validità del campo Acknowledgement.

Ausbreitung einer Infektion durch ein System Brownscher Bewegungen

In dieser Arbeit werden wir ein Modell untersuchen, welches die Ausbreitung einer Infektion beschreibt. Bei diesem Modell werden zunächst Partikel gemäß eines Poissonschen Punktprozesses auf der reellen Achse verteilt. Bis zu einem gewissen Punkt auf der reellen Achse sind alle Partikel von einer Infektion befallen. Während sich nicht infizierte Partikel nicht bewegen, folgen die infizierten Partikel den Pfaden von voneinander unabhängigen Brownschen Bewegungen und verbreitet die Infektion dabei an den Orten, welche sie betreten. Wenn sie dabei auf ein nicht infiziertes Partikel treffen, ist dieses von diesem Moment an auch infiziert und beginnt ebenfalls, dem Pfad einer Brownschen Bewegung zu folgen und die Infektion auszubreiten. Auf diese Art verschiebt sich nun der am weitesten rechts liegende Ort R_t, an dem die Infektion bereits verbreitet wurde. Wir werden mit Hilfe des subadditiven Ergodensatzes zeigen, dass sich dieser Ort mit linearer Geschwindigkeit fortbewegt. Ferner werden wir eine obere und eine untere Schranke für die Ausbreitungsgeschwindkeit angeben. Danach werden wir zeigen, dass der Prozess Regenerationszeiten hat, nämlich solche zufällige Zeiten, zu denen er eine Art Neustart unter speziellen Startbedingungen durchführt. Wir werden diese für eine weitere Charakterisierung der Ausbreitungsgeschwingkeit nutzen. Ferner erhalten wir durch die Regenerationszeiten auch einen Zentralen Grenzwertsatz für R_t und können zeigen, dass die Verteilung der infizierten Partikel aus Sicht des am weitesten rechts liegenden infizierten Ortes gegen eine invariante Verteilung konvergiert.

Random lattice models

This thesis deals with three different physical models, where each model involves a random component which is linked to a cubic lattice. First, a model is studied, which is used in numerical calculations of Quantum Chromodynamics.In these calculations random gauge-fields are distributed on the bonds of the lattice. The formulation of the model is fitted into the mathematical framework of ergodic operator families. We prove, that for small coupling constants, the ergodicity of the underlying probability measure is indeed ensured and that the integrated density of states of the Wilson-Dirac operator exists. The physical situations treated in the next two chapters are more similar to one another. In both cases the principle idea is to study a fermion system in a cubic crystal with impurities, that are modeled by a random potential located at the lattice sites. In the second model we apply the Hartree-Fock approximation to such a system. For the case of reduced Hartree-Fock theory at positive temperatures and a fixed chemical potential we consider the limit of an infinite system. In that case we show the existence and uniqueness of minimizers of the Hartree-Fock functional. In the third model we formulate the fermion system algebraically via C*-algebras. The question imposed here is to calculate the heat production of the system under the influence of an outer electromagnetic field. We show that the heat production corresponds exactly to what is empirically predicted by Joule's law in the regime of linear response.

LoneStar : Design and evaluation of an energy-efficient, disk-based archival storage system

Bandlaufwerke waren bisher die vorherrschende Technologie, um die anfallenden Datenmengen in Archivsystemen zu speichern. Mit Zugriffsmustern, die immer aktiver werden, und Speichermedien wie Festplatten die kostenmäßig aufholen, muss die Architektur vor Speichersystemen zur Archivierung neu überdacht werden. Zuverlässigkeit, Integrität und Haltbarkeit sind die Haupteigenschaften der digitalen Archivierung. Allerdings nimmt auch die Zugriffsgeschwindigkeit einen erhöhten Stellenwert ein, wenn aktive Archive ihre gesamten Inhalte für den direkten Zugriff bereitstellen. Ein band-basiertes System kann die hierfür benötigte Parallelität, Latenz und Durchsatz nicht liefern, was in der Regel durch festplattenbasierte Systeme als Zwischenspeicher kompensiert wird.rnIn dieser Arbeit untersuchen wir die Herausforderungen und Möglichkeiten ein festplattenbasiertes Speichersystem zu entwickeln, das auf eine hohe Zuverlässigkeit und Energieeffizienz zielt und das sich sowohl für aktive als auch für kalte Archivumgebungen eignet. Zuerst analysieren wir die Speichersysteme und Zugriffsmuster eines großen digitalen Archivs und präsentieren damit ein mögliches Einsatzgebiet für unsere Architektur. Daraufhin stellen wir Mechanismen vor um die Zuverlässigkeit einer einzelnen Festplatte zu verbessern und präsentieren sowie evaluieren einen neuen, energieeffizienten, zwei- dimensionalen RAID Ansatz der für „Schreibe ein Mal, lese mehrfach“ Zugriffe optimiert ist. Letztlich stellen wir Protokollierungs- und Zwischenspeichermechanismen vor, die die zugrundeliegenden Ziele unterstützen und evaluieren das RAID System in einer Dateisystemumgebung.

Mission synthesis of sine-on-random excitations for accelerated vibration qualification testing

In most real-life environments, mechanical or electronic components are subjected to vibrations. Some of these components may have to pass qualification tests to verify that they can withstand the fatigue damage they will encounter during their operational life. In order to conduct a reliable test, the environmental excitations can be taken as a reference to synthesize the test profile: this procedure is referred to as “test tailoring”. Due to cost and feasibility reasons, accelerated qualification tests are usually performed. In this case, the duration of the original excitation which acts on the component for its entire life-cycle, typically hundreds or thousands of hours, is reduced. In particular, the “Mission Synthesis” procedure lets to quantify the induced damage of the environmental vibration through two functions: the Fatigue Damage Spectrum (FDS) quantifies the fatigue damage, while the Maximum Response Spectrum (MRS) quantifies the maximum stress. Then, a new random Power Spectral Density (PSD) can be synthesized, with same amount of induced damage, but a specified duration in order to conduct accelerated tests. In this work, the Mission Synthesis procedure is applied in the case of so-called Sine-on-Random vibrations, i.e. excitations composed of random vibrations superimposed on deterministic contributions, in the form of sine tones typically due to some rotating parts of the system (e.g. helicopters, engine-mounted components, …). In fact, a proper test tailoring should not only preserve the accumulated fatigue damage, but also the “nature” of the excitation (in this case the sinusoidal components superimposed on the random process) in order to obtain reliable results. The classic time-domain approach is taken as a reference for the comparison of different methods for the FDS calculation in presence of Sine-on-Random vibrations. Then, a methodology to compute a Sine-on-Random specification based on a mission FDS is presented.

On the recurrence of random walks in Lévy random environments

This thesis investigates one-dimensional random walks in random environment whose transition probabilities might have an infinite variance. The ergodicity of the dynamical system ''from the point of view of the particle'' is proved under the assumptions of transitivity and existence of an absolutely continuous steady state on the space of the environments. We show that, if the average of the local drift over the environments is summable and null, then the RWRE is recurrent. We provide an example satisfying all the hypotheses.

Stationary states in random walks on networks

In this thesis we dealt with the problem of describing a transportation network in which the objects in movement were subject to both finite transportation capacity and finite accomodation capacity. The movements across such a system are realistically of a simultaneous nature which poses some challenges when formulating a mathematical description. We tried to derive such a general modellization from one posed on a simplified problem based on asyncronicity in particle transitions. We did so considering one-step processes based on the assumption that the system could be describable through discrete time Markov processes with finite state space. After describing the pre-established dynamics in terms of master equations we determined stationary states for the considered processes. Numerical simulations then led to the conclusion that a general system naturally evolves toward a congestion state when its particle transition simultaneously and we consider one single constraint in the form of network node capacity. Moreover the congested nodes of a system tend to be located in adjacent spots in the network, thus forming local clusters of congested nodes.

Computer Aided Surgery for Percutaneous Nephrolithotomy: Clinical Requirement Analysis and System Design

Percutaneous nephrolithotomy (PCNL) for the treatment of renal stones and other related renal diseases has proved its efficacy and has stood the test of time compared with open surgical methods and extracorporal shock wave lithotripsy. However, access to the collecting system of the kidney is not easy because the available intra-operative image modalities only provide a two dimensional view of the surgical scenario. With this lack of visual information, several punctures are often necessary which, increases the risk of renal bleeding, splanchnic, vascular or pulmonary injury, or damage to the collecting system which sometimes makes the continuation of the procedure impossible. In order to address this problem, this paper proposes a workflow for introduction of a stereotactic needle guidance system for PCNL procedures. An analysis of the imposed clinical requirements, and a instrument guidance approach to provide the physician with a more intuitive planning and visual guidance to access the collecting system of the kidney are presented.

Clinical outcome and predictors for adverse events after transcatheter aortic valve implantation with the use of different devices and access routes

Background Transcatheter aortic valve implantation (TAVI) is a treatment option for high-risk patients with severe aortic stenosis. Previous reports focused on a single device or access site, whereas little is known of the combined use of different devices and access sites as selected by the heart team. The purpose of this study is to investigate clinical outcomes of TAVI using different devices and access sites. Methods A consecutive cohort of 200 patients underwent TAVI with the Medtronic CoreValve Revalving system (Medtronic Core Valve LLC, Irvine, CA; n = 130) or the Edwards SAPIEN valve (Edwards Lifesciences LLC, Irvine, CA; n = 70) implanted by either the transfemoral or transapical access route. Results Device success and procedure success were 99% and 95%, respectively, without differences between devices and access site. All-cause mortality was 7.5% at 30 days, with no differences between valve types or access sites. Using multivariable analysis, low body mass index (<20 kg/m2) (odds ratio [OR] 6.6, 95% CI 1.5-29.5) and previous stroke (OR 4.4, 95% CI 1.2-16.8) were independent risk factors for short-term mortality. The VARC-defined combined safety end point occurred in 18% of patients and was driven by major access site complications (8.0%), life-threatening bleeding (8.5%) or severe renal failure (4.5%). Transapical access emerged as independent predictor of adverse outcome for the Valve Academic Research Consortium–combined safety end point (OR 3.3, 95% CI 1.5-7.1). Conclusion A heart team–based selection of devices and access site among patients undergoing TAVI resulted in high device and procedural success. Low body mass index and history of previous stroke were independent predictors of mortality. Transapical access emerged as a risk factor for the Valve Academic Research Consortium–combined safety end point.

Bringing results to fruition through publication: An analysis of a peer-reviewed, open access and context-focused journal's editorial practice

When it comes to helping to shape sustainable development, research is most useful when it bridges the science–implementation/management gap and when it brings development specialists and researchers into a dialogue (Hurni et al. 2004); can a peer-reviewed journal contribute to this aim? In the classical system for validation and dissemination of scientific knowledge, journals focus on knowledge exchange within the academic community and do not specifically address a ‘life-world audience’. Within a North-South context, another knowledge divide is added: the peer review process excludes a large proportion of scientists from the South from participating in the production of scientific knowledge (Karlsson et al. 2007). Mountain Research and Development (MRD) is a journal whose mission is based on an editorial strategy to build the bridge between research and development and ensure that authors from the global South have access to knowledge production, ultimately with a view to supporting sustainable development in mountains. In doing so, MRD faces a number of challenges that we would like to discuss with the td-net community, after having presented our experience and strategy as editors of this journal. MRD was launched in 1981 by mountain researchers who wanted mountains to be included in the 1992 Rio process. In the late 1990s, MRD realized that the journal needed to go beyond addressing only the scientific community. It therefore launched a new section addressing a broader audience in 2000, with the aim of disseminating insights into, and recommendations for, the implementation of sustainable development in mountains. In 2006, we conducted a survey among MRD’s authors, reviewers, and readers (Wymann et al. 2007): respondents confirmed that MRD had succeeded in bridging the gap between research and development. But we realized that MRD could become an even more efficient tool for sustainability if development knowledge were validated: in 2009, we began submitting ‘development’ papers (‘transformation knowledge’) to external peer review of a kind different from the scientific-only peer review (for ‘systems knowledge’). At the same time, the journal became open access in order to increase the permeability between science and society, and ensure greater access for readers and authors in the South. We are currently rethinking our review process for development papers, with a view to creating more space for communication between science and society, and enhancing the co-production of knowledge (Roux 2008). Hopefully, these efforts will also contribute to the urgent debate on the ‘publication culture’ needed in transdisciplinary research (Kueffer et al. 2007).