Identifizierung und funktionelle Charakterisierung neuer A-Kinase-Ankerproteine (AKAPs) im Modellorganismus C. elegans

In dieser Arbeit sollten neue Interaktionspartner der regulatorischen Untereinheit (R-UE) der Proteinkinase A (PKA) und des Modellorganismus C. elegans identifiziert und funktionell charakterisiert werden. Im Gegensatz zu Säugern (vier Isoformen), exprimiert der Nematode nur eine PKA-R-Isoform. Mittels in silico Analysen und so genannten „Pulldown“ Experimenten, wurde insbesondere nach A Kinase Ankerproteinen (AKAP) in C. elegans gesucht. Aus in silico Recherchen resultiert das rgs5 Protein als mögliches Funktionshomolog des humanen AKAP10. Rgs5 enthält eine potenzielle, amphipathische Helix (AS 421-446, SwissProt ID A9Z1K0), die in Peptide-SPOT-Arrays (durchgeführt im Biotechnologie Zentrum in Oslo, AG Prof. K. Taskén) eine Bindung an RI und RII-UE zeigt. Eine ähnliche Lokalisation von rgs5 und hAKAP10 in der Zelle, sowie vergleichende BRET² Studien, weisen auf eine mögliche Funktionshomologie zwischen AKAP10 und rgs5 hin. Die hier durchgeführten Analysen deuten darauf hin, dass es sich bei rgs5 um ein neues, klassisches AKAP mit „RII bindender Domäne“ Motiv im Modellorganismus C. elegans handelt. Basierend auf so genannten „pulldown“ Versuchen können, neben „klassischen“ AKAPs (Interaktion über amphipathische Helices), auch Interaktionspartner ohne typische Helixmotive gefunden werden. Dazu gehört auch RACK1, ein multifunktionales Protein mit 7 WD40 Domänen, das ubiquitär exprimiert wird und bereits mehr als 70 Interaktionspartner in unterschiedlichsten Signalwegen komplexiert (Adams et al., 2011). Durch BRET² Interaktionsstudien und Oberflächenplasmonresonanz (SPR) Analysen konnten hRI und kin2 als spezifische Interaktionspartner von RACK1 verifiziert werden. Untersuchungen zur Identifikation der Interaktionsflächen der beiden Proteine RACK1 und hRI zeigten im BRET² System, dass RACK1 über die WD40 Domänen 1-2 und 6-7 interagiert. Die Analyse unterschiedlicher hRI-Deletionsmutanten deutet auf die DD-Domäne im N-Terminus und zusätzlich auf eine potenzielle BH3 Domäne im C-Terminus des Proteins als Interaktionsfläche mit RACK1 hin. Die Koexpression von hRI BH3 und RACK1 zeigt einen auffälligen ein Phänotyp in Cos7 Zellen. Dieser zeichnet sich unter anderem durch eine Degradation des Zellkerns, DNA Kondensation und eine starke Vakuolisierung aus, was beides als Anzeichen für einen programmierten Zelltod interpretiert werden könnte. Erste Untersuchungen zum Mechanismus des ausgelösten Zelltods deuten auf eine Caspase unabhängige Apoptose (Paraptose) hin und einen bislang unbekannten Funktionsmechanismus der PKA hin.

Abbildung von XML-Dokumenten auf SQL:2003-konforme Datentypen

Die Auszeichnungssprache XML dient zur Annotation von Dokumenten und hat sich als Standard-Datenaustauschformat durchgesetzt. Dabei entsteht der Bedarf, XML-Dokumente nicht nur als reine Textdateien zu speichern und zu transferieren, sondern sie auch persistent in besser strukturierter Form abzulegen. Dies kann unter anderem in speziellen XML- oder relationalen Datenbanken geschehen. Relationale Datenbanken setzen dazu bisher auf zwei grundsätzlich verschiedene Verfahren: Die XML-Dokumente werden entweder unverändert als binäre oder Zeichenkettenobjekte gespeichert oder aber aufgespalten, sodass sie in herkömmlichen relationalen Tabellen normalisiert abgelegt werden können (so genanntes „Flachklopfen“ oder „Schreddern“ der hierarchischen Struktur). Diese Dissertation verfolgt einen neuen Ansatz, der einen Mittelweg zwischen den bisherigen Lösungen darstellt und die Möglichkeiten des weiterentwickelten SQL-Standards aufgreift. SQL:2003 definiert komplexe Struktur- und Kollektionstypen (Tupel, Felder, Listen, Mengen, Multimengen), die es erlauben, XML-Dokumente derart auf relationale Strukturen abzubilden, dass der hierarchische Aufbau erhalten bleibt. Dies bietet zwei Vorteile: Einerseits stehen bewährte Technologien, die aus dem Bereich der relationalen Datenbanken stammen, uneingeschränkt zur Verfügung. Andererseits lässt sich mit Hilfe der SQL:2003-Typen die inhärente Baumstruktur der XML-Dokumente bewahren, sodass es nicht erforderlich ist, diese im Bedarfsfall durch aufwendige Joins aus den meist normalisierten und auf mehrere Tabellen verteilten Tupeln zusammenzusetzen. In dieser Arbeit werden zunächst grundsätzliche Fragen zu passenden, effizienten Abbildungsformen von XML-Dokumenten auf SQL:2003-konforme Datentypen geklärt. Darauf aufbauend wird ein geeignetes, umkehrbares Umsetzungsverfahren entwickelt, das im Rahmen einer prototypischen Applikation implementiert und analysiert wird. Beim Entwurf des Abbildungsverfahrens wird besonderer Wert auf die Einsatzmöglichkeit in Verbindung mit einem existierenden, ausgereiften relationalen Datenbankmanagementsystem (DBMS) gelegt. Da die Unterstützung von SQL:2003 in den kommerziellen DBMS bisher nur unvollständig ist, muss untersucht werden, inwieweit sich die einzelnen Systeme für das zu implementierende Abbildungsverfahren eignen. Dabei stellt sich heraus, dass unter den betrachteten Produkten das DBMS IBM Informix die beste Unterstützung für komplexe Struktur- und Kollektionstypen bietet. Um die Leistungsfähigkeit des Verfahrens besser beurteilen zu können, nimmt die Arbeit Untersuchungen des nötigen Zeitbedarfs und des erforderlichen Arbeits- und Datenbankspeichers der Implementierung vor und bewertet die Ergebnisse.

Bit-Width Analysis for General Applications

It has been widely known that a significant part of the bits are useless or even unused during the program execution. Bit-width analysis targets at finding the minimum bits needed for each variable in the program, which ensures the execution correctness and resources saving. In this paper, we proposed a static analysis method for bit-widths in general applications, which approximates conservatively at compile time and is independent of runtime conditions. While most related work focus on integer applications, our method is also tailored and applicable to floating point variables, which could be extended to transform floating point number into fixed point numbers together with precision analysis. We used more precise representations for data value ranges of both scalar and array variables. Element level analysis is carried out for arrays. We also suggested an alternative for the standard fixed-point iterations in bi-directional range analysis. These techniques are implemented on the Trimaran compiler structure and tested on a set of benchmarks to show the results.

SCAPA (Spacially Addressable Protein Array) – a Novel Protein Array for the Differential Profiling of Ligand-receptor Induced Signalling

While protein microarray technology has been successful in demonstrating its usefulness for large scale high-throughput proteome profiling, performance of antibody/antigen microarrays has been only moderately productive. Immobilization of either the capture antibodies or the protein samples on solid supports has severe drawbacks. Denaturation of the immobilized proteins as well as inconsistent orientation of antibodies/ligands on the arrays can lead to erroneous results. This has prompted a number of studies to address these challenges by immobilizing proteins on biocompatible surfaces, which has met with limited success. Our strategy relates to a multiplexed, sensitive and high-throughput method for the screening quantification of intracellular signalling proteins from a complex mixture of proteins. Each signalling protein to be monitored has its capture moiety linked to a specific oligo ‘tag’. The array involves the oligonucleotide hybridization-directed localization and identification of different signalling proteins simultaneously, in a rapid and easy manner. Antibodies have been used as the capture moieties for specific identification of each signaling protein. The method involves covalently partnering each antibody/protein molecule with a unique DNA or DNA derivatives oligonucleotide tag that directs the antibody to a unique site on the microarray due to specific hybridization with a complementary tag-probe on the array. Particular surface modifications and optimal conditions allowed high signal to noise ratio which is essential to the success of this approach.

Tubular Organization of SnO₂ Nanocrystallites for Improved Lithium Ion Battery Anode Performance

Porous tin oxide nanotubes were obtained by vacuum infiltration of tin oxide nanoparticles into porous aluminum oxide membranes, followed by calcination. The porous tin oxide nanotube arrays so prepared were characterized by FE-SEM, TEM, HRTEM, and XRD. The nanotubes are open-ended, highly ordered with uniform cross-sections, diameters and wall thickness. The tin oxide nanotubes were evaluated as a substitute anode material for the lithium ion batteries. The tin oxide nanotube anode could be charged and discharged repeatedly, retaining a specific capacity of 525 mAh/g after 80 cycles. This capacity is significantly higher than the theoretical capacity of commercial graphite anode (372 mAh/g) and the cyclability is outstanding for a tin based electrode. The cyclability and capacities of the tin oxide nanotubes were also higher than their building blocks of solid tin oxide nanoparticles. A few factors accounting for the good cycling performance and high capacity of tin oxide nanotubes are suggested.

Diseño y realización de servicios de presentación en entornos gráficos.

En cada unidad didáctica precede al tít.: Técnico Superior en Desarrollo de Aplicaciones Informáticas y consta en marbetes: Formación Profesional a Distancia y Ciclo Formativo de Grado Superior

Identificación de escenarios a futuro para lograr competitividad en el sector de telefonía móvil en Colombia al año 2020

Este trabajo pretende explorar el desarrollo del sector de la telefonía móvil desde sus inicios hasta la actualidad en Colombia, con el fin de generar escenarios de futuro. Las herramientas prospectivas MicMac (Análisis Estructural Prospectivo), Smic (Sistema de Matrices de Impactos Cruzados) y la opinión de expertos líderes del sector, son la base principal para el desarrollo del trabajo. Las entidades gubernamentales, la CTR (Comisión de Regulación de Telecomunicaciones), y los líderes de los operadores del sector de telefonía móvil, entre otros, se han concientizado que la innovación es la base del éxito en este tipo de organizaciones y por eso se ha trabajado en mejorar su regulación, logrando de esta manera que el desarrollo de los productos y servicios que se ofrecen sean cada vez mejores y perjudique en menor medida al medio ambiente y a los usuarios. Este subsector de las telecomunicaciones, es el más dinámico y con mayor potencial. Sin embargo, este también es afectado por las condiciones económicas del mercado, la inestabilidad política, las importaciones y exportaciones derivadas de los tratados comerciales, entre otros temas. El escenario apuesta facilitaría la prestación de productos con tecnología de punta y servicios con la mejor cobertura y acceso posible a precios bajos.

Lecture 7 - Going Loopy

Looping while do while for Arrays indexes For each loop

Programming Principles: Collections and Iterators

In this session we look at how we can use collection objects like ArrayList as a more advanced type of array. We also introduce the idea of generics (forcing a collection to hold a particular type) and see how Java handles the autoboxing and unboxing of primitives. Finally we look at Iterators, a common design pattern for dealing with iteration over a collection.

Aspectos, consideraciones y orientaciones sobre la enseñanza del español de los negocios en Corea.

Resumen basado en el de la publicación. Memoria de master (Universidad Pablo de Olavide, 2010)

New insights on growth mechanisms of protein clusters at surfaces: an AFM and simulation study

Despite its relevance to a wide range of technological and fundamental areas, a quantitative understanding of protein surface clustering dynamics is often lacking. In inorganic crystal growth, surface clustering of adatoms is well described by diffusion-aggregation models. In such models, the statistical properties of the aggregate arrays often reveal the molecular scale aggregation processes. We investigate the potential of these theories to reveal hitherto hidden facets of protein clustering by carrying out concomitant observations of lysozyme adsorption onto mica surfaces, using atomic force microscopy. and Monte Carlo simulations of cluster nucleation and growth. We find that lysozyme clusters diffuse across the substrate at a rate that varies inversely with size. This result suggests which molecular scale mechanisms are responsible for the mobility of the proteins on the substrate. In addition the surface diffusion coefficient of the monomer can also be extracted from the comparison between experiments and simulations. While concentrating on a model system of lysozyme-on-mica, this 'proof of concept' study successfully demonstrates the potential of our approach to understand and influence more biomedically applicable protein-substrate couples.

Development of multi-electrode array screening for anticonvulsants in acute rat brain slices

The acute hippocampal brain slice preparation is an important in vitro screening tool for potential anticonvulsants. Application of 4-aminopyridine (4-AP) or removal of external Mg2+ ions induces epileptiform bursting in slices which is analogous to electrical brain activity seen in status epilepticus states. We have developed these epileptiform models for use with multi-electrode arrays (MEAs), allowing recording across the hippocampal slice surface from 59 points. We present validation of this novel approach and analyses using two anticonvulsants, felbamate and phenobarbital, the effects of which have already been assessed in these models using conventional extracellular recordings. In addition to assessing drug effects on commonly described parameters (duration, amplitude and frequency), we describe novel methods using the MEA to assess burst propagation speeds and the underlying frequencies that contribute to the epileptiform activity seen. Contour plots are also used as a method of illustrating burst activity. Finally, we describe hitherto unreported properties of epileptiform bursting induced by 100M4-AP or removal of external Mg2+ ions. Specifically, we observed decreases over time in burst amplitude and increase over time in burst frequency in the absence of additional pharmacological interventions. These MEA methods enhance the depth, quality and range of data that can be derived from the hippocampal slice preparation compared to conventional extracellular recordings. It may also uncover additional modes of action that contribute to anti-epileptiform drug effects

Modelling habitat selection and distribution of the critically endangered Jerdon's courser Rhinoptilus bitorquatus in scrub jungle: an application of a new tracking method

1. Jerdon's courser Rhinoptilus bitorquatus is a nocturnally active cursorial bird that is only known to occur in a small area of scrub jungle in Andhra Pradesh, India, and is listed as critically endangered by the IUCN. Information on its habitat requirements is needed urgently to underpin conservation measures. We quantified the habitat features that correlated with the use of different areas of scrub jungle by Jerdon's coursers, and developed a model to map potentially suitable habitat over large areas from satellite imagery and facilitate the design of surveys of Jerdon's courser distribution. 2. We used 11 arrays of 5-m long tracking strips consisting of smoothed fine soil to detect the footprints of Jerdon's coursers, and measured tracking rates (tracking events per strip night). We counted the number of bushes and trees, and described other attributes of vegetation and substrate in a 10-m square plot centred on each strip. We obtained reflectance data from Landsat 7 satellite imagery for the pixel within which each strip lay. 3. We used logistic regression models to describe the relationship between tracking rate by Jerdon's coursers and characteristics of the habitat around the strips, using ground-based survey data and satellite imagery. 4. Jerdon's coursers were most likely to occur where the density of large (>2 m tall) bushes was in the range 300-700 ha(-1) and where the density of smaller bushes was less than 1000 ha(-1). This habitat was detectable using satellite imagery. 5. Synthesis and applications. The occurrence of Jerdon's courser is strongly correlated with the density of bushes and trees, and is in turn affected by grazing with domestic livestock, woodcutting and mechanical clearance of bushes to create pasture, orchards and farmland. It is likely that there is an optimal level of grazing and woodcutting that would maintain or create suitable conditions for the species. Knowledge of the species' distribution is incomplete and there is considerable pressure from human use of apparently suitable habitats. Hence, distribution mapping is a high conservation priority. A two-step procedure is proposed, involving the use of ground surveys of bush density to calibrate satellite image-based mapping of potential habitat. These maps could then be used to select priority areas for Jerdon's courser surveys. The use of tracking strips to study habitat selection and distribution has potential in studies of other scarce and secretive species.

Mid-infrared filters for astronomical and remote sensing instrumentation

As improvements to the optical design of spectrometer and radiometer instruments evolve with advances in detector sensitivity, use of focal plane detector arrays and innovations in adaptive optics for large high altitude telescopes, interest in mid-infrared astronomy and remote sensing applications have been areas of progressive research in recent years. This research has promoted a number of developments in infrared coating performance, particularly by placing increased demands on the spectral imaging requirements of filters to precisely isolate radiation between discrete wavebands and improve photometric accuracy. The spectral design and construction of multilayer filters to accommodate these developments has subsequently been an area of challenging thin-film research, to achieve high spectral positioning accuracy, environmental durability and aging stability at cryogenic temperatures, whilst maximizing the far-infrared performance. In this paper we examine the design and fabrication of interference filters in instruments that utilize the mid-infrared N-band (6-15 µm) and Q-band (16-28 µm) atmospheric windows, together with a rationale for the selection of materials, deposition process, spectral measurements and assessment of environmental durability performance.