Untersuchungen zur Lokalisation und Expression des respiratorischen Proteins Neuroglobin bei Säugetieren

In der vorliegenden Arbeit wurde Neuroglobin (Ngb), ein evolutiv altes und in Metazoen konserviertes respiratorisches Protein, funktionell untersucht. Mittels des induzierbaren Tet on / Tet off Systems wurde Ngb ektopisch in der murinen Leber und im Gehirn überexprimiert. Die Transkriptome von Leber und Gehirnregionen Ngb-transgener Mäuse wurden mittels Microarrays und RNA-Seq im Vergleich zum Wildtyp analysiert, um Auswirkungen der Ngb-Überexpression zu ermitteln. Die Transkriptom-Analyse in Leber und Gehirn zeigte eine nur geringe Anzahl differenziell regulierter Gene und Stoffwechselwege nach Ngb-Überexpression. Ngb transgene Mäuse wurden CCl4-induziertem ROS-Stress ausgesetzt und die Leberfunktion untersucht. Zudem wurden primäre Hepatozyten-Kulturen etabliert und in diesen in vitro die extrinsische Apoptose induziert. Die Stressversuche zeigten: (i) Die Ngb-Überexpression hat keine protektive Wirkung in der Leber in vivo. (ii) In Leberzellen in vitro hingegen verminderte eine Ngb-Überexpression effizient die Aktivierung der apoptotischen Kaskade. Eine protektive Wirkung von Ngb ist vermutlich von betrachtetem Gewebe und dem verwendeten Stressor abhängig und keine generelle, selektierte Funktion des Proteins.rnWeiterhin wurde eine Ngb-KnockOut-Mauslinie mit einem LacZ-KnockIn-Genotyp etabliert. Hierbei zeigten die KO-Mäuse keinen offensichtlichen Phänotyp in ihrer Entwicklung, Fortpflanzung und Retina-Funktion. Unter Verwendung des LacZ-Knockin-Konstrukts konnten kontrovers diskutierte Ngb-Expressionsorte im adulten Mausgehirn (Hippocampus, Cortex und Cerebellum) sowie in Testes experimentell bestätigt werden. Parallel wurden öffentlich verfügbare RNA-Seq Datensätze ausgewertet, um die regionale Ngb-Expression systematisch ohne Antikörper-assoziierte Spezifitätsprobleme zu charakterisieren. Eine basale Ngb-Expression (RPKM: ~1-5) wurde im Hippocampus, Cortex und Cerebellum, sowie in Retina und Testes ermittelt. Eine 20-40fach höhere, starke Expression (RPKM: ~160) wurde im Hypothalamus bzw. im Hirnstamm nachgewiesen. Die „digitale“ Expressionsuntersuchung wurde mittels qRT-PCR und Western Blot bestätigt. Dieses Expressionsprofil von Ngb in der Maus weist auf eine besondere funktionelle Bedeutung von Ngb im Hypothalamus hin. Eine Funktion von Ngb in der Sauerstoffversorgung der Retina und eine generelle Funktion von Ngb in der Protektion von Neuronen sind mit dem beobachteten Expressionsspektrum weniger gut vereinbar.

Privacy Calculus on Social Networking Sites: Explorative Evidence from Germany and USA

Worldwide social networks, like Facebook, face fierce competition from local platforms when expanding globally. To remain attractive social network providers need to encourage user self-disclosure. Yet, little research exists on how cultural differences impact selfdisclosure on these platforms. Addressing this gap, this study explores the differences in perceptions of disclosure-relevant determinants between German and US users. Survey of Facebook members indicates that German users expect more damage and attribute higher probability to privacy-related violations. On the other hand, even though American users show higher level of privacy concern, they extract more benefits from their social networking activities, have more trust in the service provider and legal assurances as well as perceive more control. These factors may explain a higher level of self-disclosure indicated by American users. Our results provide relevant insights for the social network providers who can adjust their expansion strategy with regard to cultural differences.

Coercive, Normative, and Mimetic Influences on the Assimilation of BCM in Outsourcing Relationships

Today many business processes are based on IT systems. These systems are exposed to different threats, which may lead to failures of critical business pro-cesses. Thus, enterprises prepare themselves against threats and failures of critical IT systems by means of Business Continuity Management (BCM). The phe-nomenon of outsourcing introduces a new dimension to BCM. In an outsourcing relationship the client organization is still responsible for the continuity of its processes but does not have full control over the implemented business continuity measures. In this paper we build a research model based on institutional and assimilation theories to describe and explain how and why BCM is assimilated in outsourcing relationships. In our case studies we found evidence that primarily coercive and normative pressures influence the assimilation of BCM in outsourcing relationships and support the explanation of variation across enterprises. Mimetic pressures seem to influence the assimilation but do not explain variations.

The C-terminal Domain of p21 Inhibits Nucleotide Excision Repair In Vitro and In Vivo

The protein p21Cip1, Waf1, Sdi1 is a potent inhibitor of cyclin-dependent kinases (CDKs). p21 can also block DNA replication through its interaction with the proliferating cell nuclear antigen (PCNA), which is an auxiliary factor for polymerase δ. PCNA is also implicated in the repair resynthesis step of nucleotide excision repair (NER). Previous studies have yielded contradictory results on whether p21 regulates NER through its interaction with PCNA. Resolution of this controversy is of interest because it would help understand how DNA repair and replication are regulated. Hence, we have investigated the effect of p21 on NER both in vitro and in vivo using purified fragments of p21 containing either the CDK-binding domain (N terminus) or the PCNA binding domain (C terminus) of the protein. In the in vitro studies, DNA repair synthesis was measured in extracts from normal human fibroblasts using plasmids damaged by UV irradiation. In the in vivo studies, we used intact and permeabilized cells. The results show that the C terminus of the p21 protein inhibits NER both in vitro and in vivo. These are the first in vivo studies in which this question has been examined, and we demonstrate that inhibition of NER by p21 is not merely an artificial in vitro effect. A 50% inhibition of in vitro NER occurred at a 50:1 molar ratio of p21 C-terminus fragment to PCNA monomer. p21 differentially regulates DNA repair and replication, with repair being much less sensitive to inhibition than replication. Our in vivo results suggest that the inhibition occurs at the resynthesis step of the repair process. It also appears that preassembly of PCNA at repair sites mitigates the inhibitory effect of p21. We further demonstrate that the inhibition of DNA repair is mediated via binding of p21 to PCNA. The N terminus of p21 had no effect on DNA repair, and the inhibition of DNA repair by the C terminus of p21 was relieved by the addition of purified PCNA protein.

Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice.

The tet regulatory system in which doxycycline (dox) acts as an inducer of specifically engineered RNA polymerase II promoters was transferred into transgenic mice. Tight control and a broad range of regulation spanning up to five orders of magnitude were monitored dependent on the dox concentration in the water supply of the animals. Administration of dox rapidly induces the synthesis of the indicator enzyme luciferase whose activity rises over several orders of magnitude within the first 4 h in some organs. Induction is complete after 24 h in most organs analyzed. A comparable regulatory potential was revealed with the tet regulatory system where dox prevents transcription activation. Directing the synthesis of the tetracycline-controlled transactivator (tTA) to the liver led to highly specific regulation in hepatocytes where, in presence of dox, less than one molecule of luciferase was detected per cell. By contrast, a more than 10(5)-fold activation of the luciferase gene was observed in the absence of the antibiotic. This regulation was homogeneous throughout but stringently restricted to hepatocytes. These results demonstrate that both tetracycline-controlled transcriptional activation systems provide genetic switches that permit the quantitative control of gene activities in transgenic mice in a tissue-specific manner and, thus, suggest possibilities for the generation of a novel type of conditional mutants.

Third report on survey of electric utility applications of digital computers.

At head of title: S-109-A.

Second report on survey of electric utility applications of digital computers.

At head of title: S-109.

Power system sensitivity analysis for probabilistic small signal stability assessment in a deregulated environment

Deregulations and market practices in power industry have brought great challenges to the system planning area. In particular, they introduce a variety of uncertainties to system planning. New techniques are required to cope with such uncertainties. As a promising approach, probabilistic methods are attracting more and more attentions by system planners. In small signal stability analysis, generation control parameters play an important role in determining the stability margin. The objective of this paper is to investigate power system state matrix sensitivity characteristics with respect to system parameter uncertainties with analytical and numerical approaches and to identify those parameters have great impact on system eigenvalues, therefore, the system stability properties. Those identified parameter variations need to be investigated with priority. The results can be used to help Regional Transmission Organizations (RTOs) and Independent System Operators (ISOs) perform planning studies under the open access environment.

Effect of different preparation routes on the structure and properties of rigid polyurethane-layered silicate nanocomposites

To identify the effect of reactive preparation on the structure and properties of rigid polyurethane (PU)layered silicate nanocomposite, a range of nanocomposites were prepared by combining the various precursors in different sequences. The morphology of the samples was characterized by XRD and TEM. Tensile properties and dynamic mechanical thermal properties were measured. The reactions between the layered silicates and PU precursors were monitored via FTIR to gain an understanding of the participation of nanofiller in the polymerization reaction, and the impact of this on system stoichiometry. The XRD and TEM results provided evidence that morphology can differ significantly if different synthesis methods are used. However, the mechanical properties are dominated by the stoichiometry imbalance induced by the addition of the layered silicates. (c) 2006 Wiley Periodicals, Inc.

Strategic development of transport systems: a study of the physical constraints on planning processes

Investment in transport infrastructure can be highly sensitive to uncertainty. The scale and lead time of strategic transport programmes are such that they require continuing policy support and accurate forecasting. Delay, cost escalation and abandonment of projects often result if these conditions are not present. In Part One the physical characteristics of infrastructure are identified as a major constraint on planning processes. The extent to which strategies and techniques acknowledge these constraints is examined. A simple simulation model is developed to evaluate the effects on system development of variations in the scale and lead time of investments. In Part Two, two case studies of strategic infrastructure investment are analysed. The absence of a policy consensus for airport location was an important factor in the delayed resolution of the Third London Airport issue. In London itself, the traffic and environmental effects of major highway investment ultimately resulted in the abandonment of plans to construct urban motorways. In both cases, the infrastructure implications of alternative strategies are reviewed with reference to the problems of uncertainty. In conclusion, the scale of infrastructure investment is considered the most important of the constraints on the processes of transport planning. Adequate appraisal of such constraints may best be achieved by evaluation more closely aligned to policy objectives.

Cross-linking of cellular proteins by tissue transglutaminase during necrotic cell death:a mechanism for maintaining tissue integrity

Tissue transglutaminase (tTG) is a Ca2+-dependent enzyme which cross-links proteins via e(g-glutamyl)lysine bridges. There is increasing evidence that tTG is involved in wound repair and tissue stabilization, as well as in physiological mechanisms leading to cell death. To investigate the role of this enzyme in tissue wounding leading to loss of Ca2+ homoeostasis, we initially used a model involving electroporation to reproduce cell wounding under controlled conditions. Two cell models were used whereby tTG expression is regulated either by antisense silencing in ECV 304 cells or by using transfected Swiss 3T3 cells in which tTG expression is under the control of the tet regulatory system. Using these cells, loss of Ca2+ homoeostasis following electroporation led to a tTG-dependent formation of highly cross-linked proteinaceous shells from intracellular proteins. Formation of these structures is dependent on elevated intracellular Ca2+, but it is independent of intracellular proteases and is near maximal after only 20min post-wounding. Using labelled primary amines as an indicator of tTG activity within these 'wounded cells', we demonstrate that tTG modifies a wide range of proteins that are present in both the perinuclear and intranuclear spaces. The demonstration of entrapped DNA within these shell structures, which showed limited fragmentation, provides evidence that the high degree of transglutaminase cross-linking results in the prevention of DNA release, which may serve to dampen any subsequent inflammatory response. Comparable observations were shown when monolayers of cells were mechanically wounded by scratching. In this second model of cell wounding, redistribution of tTG activity to the extracellular matrix was also demonstrated, an effect which may serve to stabilize tissues post-trauma, and thus contribute to the maintenance of tissue integrity.

Multipurpose portable system for environmental, industrial, and medical applications

This dissertation is about the research carried on developing an MPS (Multipurpose Portable System) which consists of an instrument and many accessories. The instrument is portable, hand-held, and rechargeable battery operated, and it measures temperature, absorbance, and concentration of samples by using optical principles. The system also performs auxiliary functions like incubation and mixing. This system can be used in environmental, industrial, and medical applications. ^ Research emphasis is on system modularity, easy configuration, accuracy of measurements, power management schemes, reliability, low cost, computer interface, and networking. The instrument can send the data to a computer for data analysis and presentation, or to a printer. ^ This dissertation includes the presentation of a full working system. This involved integration of hardware and firmware for the micro-controller in assembly language, software in C and other application modules. ^ The instrument contains the Optics, Transimpedance Amplifiers, Voltage-to-Frequency Converters, LCD display, Lamp Driver, Battery Charger, Battery Manager, Timer, Interface Port, and Micro-controller. ^ The accessories are a Printer, Data Acquisition Adapter (to transfer the measurements to a computer via the Printer Port and expand the Analog/Digital conversion capability), Car Plug Adapter, and AC Transformer. This system has been fully evaluated for fault tolerance and the schemes will also be presented. ^

Event mining for system and service management

Modern IT infrastructures are constructed by large scale computing systems and administered by IT service providers. Manually maintaining such large computing systems is costly and inefficient. Service providers often seek automatic or semi-automatic methodologies of detecting and resolving system issues to improve their service quality and efficiency. This dissertation investigates several data-driven approaches for assisting service providers in achieving this goal. The detailed problems studied by these approaches can be categorized into the three aspects in the service workflow: 1) preprocessing raw textual system logs to structural events; 2) refining monitoring configurations for eliminating false positives and false negatives; 3) improving the efficiency of system diagnosis on detected alerts. Solving these problems usually requires a huge amount of domain knowledge about the particular computing systems. The approaches investigated by this dissertation are developed based on event mining algorithms, which are able to automatically derive part of that knowledge from the historical system logs, events and tickets. ^ In particular, two textual clustering algorithms are developed for converting raw textual logs into system events. For refining the monitoring configuration, a rule based alert prediction algorithm is proposed for eliminating false alerts (false positives) without losing any real alert and a textual classification method is applied to identify the missing alerts (false negatives) from manual incident tickets. For system diagnosis, this dissertation presents an efficient algorithm for discovering the temporal dependencies between system events with corresponding time lags, which can help the administrators to determine the redundancies of deployed monitoring situations and dependencies of system components. To improve the efficiency of incident ticket resolving, several KNN-based algorithms that recommend relevant historical tickets with resolutions for incoming tickets are investigated. Finally, this dissertation offers a novel algorithm for searching similar textual event segments over large system logs that assists administrators to locate similar system behaviors in the logs. Extensive empirical evaluation on system logs, events and tickets from real IT infrastructures demonstrates the effectiveness and efficiency of the proposed approaches.^

Validating Scanned Foot Images and Designing Customized Insoles on the Cloud

People with foot problems need special healthcare: foot care. Customized insoles can provide this care. They are inserts that are placed in the shoes. They correct biomechanical and postural inaccuracies in foot. Insole production contains four phases: foot image scanning, image validation, insole design and insole manufacturing. Currently, image scanning and validation is separated in location and time, i.e. podiatrists take images and insole designers validate them at different location and at different time. A cloud-based solution, the CloudSME one-stop shop simulation platform, enables remote access to image validation and insole design service deployed and running on the Cloud. The remote access allows podiatrists validating scanned image while the patient is in their offices. The simulation platform also supports remote design of customized insoles.

Carrier transfer in the optical recombination of quantum dots

We report a study of dynamic effects detected in the time-resolved emission from quantum dot ensembles. Experimental procedures were developed to search for common behaviors found in quantum dot systems independently of their composition: three quantum dot samples were experimentally characterized. Systems with contrasting interdot coupling are compared and their sensitivity to the excitation energy is analyzed. Our experimental results are compared and contrasted with other results available in literature. The optical recombination time dependence on system parameters is derived and compared to the experimental findings. We discuss the effects of occupation of the ground state in both valence and conduction bands of semiconductor quantum dots in the dynamics of the system relaxation as well as the nonlinear effects.