A protocol describing the genetic correction of somatic human cells and subsequent generation of IPS cell

The generation of patient-specific induced pluripotent stem cells (iPSCPSCPSCs) offers unprecedented opportunities for modeling and treating human disease. In combination with gene therapy, the iPSCPSCPSC technology can be used to generate disease-free progenitor cells of potential interest for autologous cell therapy. We explain a protocol for the reproducible generation of genetically corrected iPSCPSCPSCs starting from the skin biopsies of Fanconi anemia patients using retroviral transduction with OCT4, SOX2 and KLF4. Before reprogramming, the fibroblasts and/or keratinocytes of the patients are genetically corrected with lentiviruses expressing FANCA. The same approach may be used for other diseases susceptible to gene therapy correction. Genetically corrected, characterized lines of patient-specific iPSCPSCPSCs can be obtained in 4–5 months.

The dynamics of the Swiss electricity market : three essays

Executive Summary Electricity is crucial for modern societies, thus it is important to understand the behaviour of electricity markets in order to be prepared to face the consequences of policy changes. The Swiss electricity market is now in a transition stage from a public monopoly to a liberalised market and it is undergoing an "emergent" liberalisation - i.e. liberalisation taking place without proper regulation. The withdrawal of nuclear capacity is also being debated. These two possible changes directly affect the mechanisms for capacity expansion. Thus, in this thesis we concentrate on understanding the dynamics of capacity expansion in the Swiss electricity market. A conceptual model to help understand the dynamics of capacity expansion in the Swiss electricity market is developed an explained in the first essay. We identify a potential risk of imports dependence. In the second essay a System Dynamics model, based on the conceptual model, is developed to evaluate the consequences of three scenarios: a nuclear phase-out, the implementation of a policy for avoiding imports dependence, and the combination of both. We conclude that the Swiss market is not well prepared to face unexpected changes of supply and demand, and we identify a risk of imports dependence, mainly in the case of a nuclear phase-out. The third essay focus on the opportunity cost of hydro-storage power generation, one of the main generation sources in Switzerland. We use and extended version of our model to test different policies for assigning an opportunity cost to hydro-storage power generation. We conclude that the preferred policies are different for different market participants and depend on market structure.

Improving electricity market price scenarios by means of forecasting factor models

In liberalized electricity markets, generation Companies must build an hourly bidthat is sent to the market operator. The price at which the energy will be paid is unknown during the bidding process and has to be forecast. In this work we apply forecasting factor models to this framework and study its suitability.

Rapid cohort generation and analysis of disease spectrum of large animal model of cone dystrophy.

Large animal models are an important resource for the understanding of human disease and for evaluating the applicability of new therapies to human patients. For many diseases, such as cone dystrophy, research effort is hampered by the lack of such models. Lentiviral transgenesis is a methodology broadly applicable to animals from many different species. When conjugated to the expression of a dominant mutant protein, this technology offers an attractive approach to generate new large animal models in a heterogeneous background. We adopted this strategy to mimic the phenotype diversity encounter in humans and generate a cohort of pigs for cone dystrophy by expressing a dominant mutant allele of the guanylate cyclase 2D (GUCY2D) gene. Sixty percent of the piglets were transgenic, with mutant GUCY2D mRNA detected in the retina of all animals tested. Functional impairment of vision was observed among the transgenic pigs at 3 months of age, with a follow-up at 1 year indicating a subsequent slower progression of phenotype. Abnormal retina morphology, notably among the cone photoreceptor cell population, was observed exclusively amongst the transgenic animals. Of particular note, these transgenic animals were characterized by a range in the severity of the phenotype, reflecting the human clinical situation. We demonstrate that a transgenic approach using lentiviral vectors offers a powerful tool for large animal model development. Not only is the efficiency of transgenesis higher than conventional transgenic methodology but this technique also produces a heterogeneous cohort of transgenic animals that mimics the genetic variation encountered in human patients.

"Next-generation" Sequencing (NGS): The new genomic revolution

In the past 5 years "Next-generation" Sequencing (NGS) technologies have transformed genomics by delivering fast, inexpensive and accurate genomeinformation changing the way we think about scientific approaches in basic,applied and clinical research. The inexpensive production of large volumes ofsequence data is the main advantage over the automated Sanger method,making this new technology useful for many applications. In this chapter, a brieftechnical review of NGS technologies is given, along with the keys to NGSsuccess and a broad range of applications for NGS technologies.

Circulating miRNAs: a new generation of anti-doping biomarkers

MicroRNAs (miRNAs) are small non-coding RNAs that regulate a variety of biological processes. Cell-free miRNAs detected in blood plasma are used as specific and sensitive markers of physiological processes and some diseases. Circulating miRNAs are highly stable in body fluids, for example plasma. Therefore, profiles of circulating miRNAs have been investigated for potential use as novel, non-invasive anti-doping biomarkers. This review describes the biological mechanisms underlying the variation of circulating miRNAs, revealing that they have great potential as a new class of biomarker for detection of doping substances. The latest developments in extraction and profiling technology, and the technical design of experiments useful for anti-doping, are also discussed. Longitudinal measurements of circulating miRNAs in the context of the athlete biological passport are proposed as an efficient strategy for the use of these new markers. The review also emphasizes potential challenges for the translation of circulating miRNAs from research into practical anti-doping applications.

Technology-enhanced education for Millennials: how the Information Society is changing the way of learning

The Information Society has provided the context for the development of a new generation, known as the Millennials, who are characterized by their intensive use of technologies in everyday life. These features are changing the way of learning, prompting educational institutions to attempt to better adapt to youngneeds by incorporating technologies into education. Based on this premise, wehave reviewed the prominent reports of the integration of ICT into education atdifferent levels with the aim of evidencing how education is changing, and willchange, to meet the needs of Millennials with ICT support. The results show thatmost of the investments have simply resulted in an increase of computers andaccess to the Internet, with teachers reproducing traditional approaches to education and e-learning being seen as complementary to face-to-face education.While it would seem that the use of ICT is not revolutionizing learning, it isfacilitating the personalization, collaboration and ubiquity of learning.

Beyond the Net Generation debate: A comparison between digital learners in face-to-face and virtual universities

This paper is based on the hypothesis that the use of technology to support learning is not related to whether a student belongs to the Net Generation, but that it is mainly influenced by the teaching model. The study compares behaviour and preferences towards ICT use in two groups of university students: face-to-face students and online students. A questionnaire was applied to asample of students from five universities with different characteristics (one offers online education and four offer face-to-face education with LMS teaching support).

Investment Determinants and Future Shape of Russia's Power Generation

The reformation of power sector is still in the process of development. The present day situation in Russian electricity power market ischaracterized as transitional period: competitive electricity market is forming, new companies are being created and the power of government regulation is decreasing. The main aim of the reformation is to attract much-needed private investments to the power sector. The electricity consumption increases very rapidly and power sector has to cope with high demand. The goal of this master's thesis is to analyze the nowadays situation in Russian power sector, such as generation structure, condition of electricity networks, electricity price formation for end-users, shape of fuel sector and investments risks and attraction.The final result of this work is creation of scenario of Russian power sector future shape and analysis of the present day situation.

Scenarios for electricity power sector development in the North-West of Russia

Focus of this thesisis made on development of electricity sector of Russian North-West. The objective was to determine the most likely scenarios for development of the most critical (western) part of Interregional Power System of North-West, from where the most part of Russian electricity exports to the countries of European Union take place. For this purpose all the involved sides were analyzed: generation, transmission system and electricity consumption in different regions of Russian North-West. The analysis was performed through investigation of existing generation andtransmission capacities and plans for their development to be performed by the generation and transmission companies operating in the region. Principles of Russian electricity sector restructuring and electricity market design are also discussed as well as factors that may influence on future electricity price in the region.

De novo T cell generation and cell cycle analysis of CD4 and CD8 T cells during HIV-disease

RESUME POUR UN LARGE PUBLIC Parmi les globules blancs, les lymphocytes T 004 jouent un rôle primordial dans la coordination de la réponse immunitaire contre les pathogènes et les lymphocytes T CD8 dans leur élimination. Lors d'une infection par le virus de l'immunodéficience humaine (VIH-1), non seulement les cellules T CD4 sont les principales cibles d'infections, mais aussi elles disparaissent progressivement tout au long de la maladie. Ce phénomène, appelé aussi épuisement des lymphocytes T CD4, est la principale cause provoquant le Syndrome d'Immunodéficience Acquise (SIDA). Malgré de grands efforts de recherche, nous ne sommes toujours pas en mesure de dire si ce phénomène est dû à un défaut dans la production de nouvelles cellules ou à une destruction massive de cellules en circulation. Dans cette étude, nous nous proposions, dans un premier temps, de comparer la production de nouvelles cellules T CD4 et CD8 chez des individus VIH-négatifs et positifs. Les cellules nouvellement produites portent un marqueur commun que l'on appelle TREC et qui est facilement mesurable. En considérant des paramètres cliniques, nous étions en mesure de déterminer le niveau de TRECs de cellules T CD4 et CD8 dans différentes phases de la maladie. De là, nous avons pu déterminer que le niveau de TREC est toujours plus bas dans les cellules T CD8 de patients VIH-positifs comparativement à notre groupe contrôle. Nous avons pu déterminer par une analyse ultérieure que cette différence est due à une forte prolifération de ces cellules chez les patients VIH-positifs, ce qui a pour effet de diluer ce marqueur. En revanche, la production de nouvelles cellules T CD4 chez des patients VIH-positifs est accentuée lors de la phase précoce de la maladie et largement réprimée lors de la phase tardive. Dans un second temps, nous avons effectué une analyse à grande échelle de l'expression de gènes associés à la division cellulaire sur des lymphocytes T CD4 et CD8 d'individus VIH-¬positifs et négatifs, avec comme contrôle des cellules proliférant in vitro. De cette étude, nous avons pu conclure que les cellules T CD8 de patients VIH-positifs étaient en état de prolifération, alors que les lymphocytes T CD4 présentaient des défauts majeurs conduisant à un arrêt de la division cellulaire. Nos résultats montrent que la capacité à produire de nouvelles cellules chez des patients VIH¬positifs reste active longtemps pendant la maladie, mais que l'incapacité des cellules T CD4 à proliférer peut enrayer la reconstitution immunitaire chez ces individus. ABSTRACT The hallmark of HIV-1 infection is the depletion of CD4 T cells. Despite extensive investigation, the mechanisms responsible for the loss of CD4 T cells have been elucidated only partially. In particular, it remains controversial whether CD4 T cell depletion results from a defect in T cell production or from a massive peripheral destruction. In this study, de novo T cell generation has been investigated by measuring T cell receptor rearrangement excision circles (TRECs) on large cohorts of HIV-negative (N=120) and HIV-1 infected (N=298) individuals. Analysis of TREC levels was performed in HIV-infected subjects stratified by the stage of HIV disease based on CD4 T cell counts (early: >500 CD4 T cells/µl; intermediate: <500>200; late: <200) and by age (20 to 60 years, n = 259). Our data show that TREC levels in CD8 T cells were significantly lower in HIV-infected subjects at any stage of disease compared to the control group. In contrast, TREC levels in CD4 T cells were significantly higher in HIV-infected subjects at early stages disease while no significant differences were observed at intermediate stages of the disease and were severely reduced only at late stages of disease. To investigate further the status of cell cycle in peripheral CD4 and CD8 T cells in HIV-1 infections, we determined the pattern of gene expression with the microarray technology. In particular, CD4 and CD8 T cells of HIV-1 infected and HIV-negative subjects were analysed by Cell Cycle cDNA expression array. The patterns of gene expression were compared to in vitro stimulated CD4 and CD8 T cells and this analysis showed that CD8 T cells of HIV-1 infected subjects had a pattern of gene expression very similar to that of in vitro stimulated CD8 T cells thus indicating ongoing cell cycling. In contrast, CD4 T cells of HIV-1 infected subjects displayed a complex pattern of gene expression. In fact, CD4 T cells expressed high levels of genes typically associated with cell activation, but low levels of cell cycle genes. Therefore, these results indicated that activated CD4 T cells of HIV-1 infected subjects were in cell cycle arrest. Taking together these results indicate that thymus function is preserved for long time during HIV- 1 infection and the increase observed in early stage disease may represent a compensatory mechanism to the depletion of CD4 T cells. However, we provide evidence for a cell cycle arrest of peripheral CD4 T cells that may prevent potentially the replenishment of CD4 T cells. RESUME Les mécanismes responsables de la perte des lymphocytes T CD4 lors de l'infection pas VIH n'ont été élucidés que partiellement. Nous ne savons toujours pas si l'épuisement des lymphocytes T CD4 résulte d'un défaut dans la production de cellules ou d'une destruction périphérique massive. Dans cette étude, la production de cellules T a été étudiée en mesurant les cercles d'excision générés lors du réarrangement du récepteur au cellules T (TRECs) chez des individus VIH-négatifs (N=120) et VIH-1 positifs (N=298). L'analyse des niveaux de TREC a été faite chez sujets HIV-infectés en considérant les phases de la maladie sur la base des comptes CD4 (phase précoce: > 500 cellules CD4/µl; intermédiaire: < 500>200; tardive: < 200) et par âge. Nos données démontrent que les niveaux de TRECs des cellules T CD8 étaient significativement plus bas chez les sujets VIH-1 infectés, à tous les stades de la maladie comparativement au groupe contrôle. En revanche, les niveaux de TRECs des cellules T CD4 étaient significativement plus élevés chez les sujets VIH-1 infectés durant la phase précoce de la maladie, tandis qu'aucune différence significative n'était observée durant la phase intermédiaire et étaient très réduits dans la phase tardive. Dans une deuxième partie, nous avons utilisé la technique des biopuces à d'ADN complémentaire pour analyser la régulation du cycle cellulaire chez les lymphocytes T CD4 et CD8 périphériques lors d'une infection au VIH-1. Des profils d'expression ont été déterminés et comparés à ceux de cellules T CD4 et CD8 stimulées in vitro, démontrant que les cellules T CD8 des sujets VIH-positifs avaient un profil d'expression très semblable à celui des cellules stimulées in vitro en prolifération. En revanche, les lymphocytes T CD4 des sujets VIH-1 positifs avaient un profil d'expression de gène plus complexe. En fait, leur profil montrait une sur- expression de gènes associés à une activation cellulaire, mais une sous-expression de ceux induisant une division. Ainsi, ces résultats indiquent que les lymphocytes T CD4 d'individus VIH-positifs présentent des dérégulations qui conduisent à un arrêt du cycle cellulaire. Ces résultats montrent que la fonction thymique est préservée longtemps pendant l'infection au VIH-1 et que l'augmentation de la quantité de TRECs dans la phase précoce de la maladie peut représenter un mécanisme compensatoire à l'épuisement des cellules T CD4. Cependant, nous démontrons aussi un clair dysfonctionnement du cycle cellulaire chez les cellules T CD4 d'individus infectés par VIH-1 ce qui peut enrayer la reconstitution du système immunitaire.

New telecom services over next generation SDH

Our main aim in this report is to use Next generation SDH to solve the problem associated with the new telecom services. We have tried to analyze the different services and in this way identified some drawbacks which can be seen as hindrances in supporting these services. In this thesis we will first try to have idea of the past SDH technology and how the next generation SDH came into effect overriding the drawbacks of the past SDH technology. Our main concern throughout the report will be the way we can use next generation SDH to provide quality telecommunication services. In the section dealing with the telecommunication services through next generation SDH we will consider how we can transport Ethernet services through the Next generation SDH and what are the benefits to the customer and the service provider in using next generation SDH as a carrier. We will also see to improve to the ATM services through Next generation SDH. And finally towards the end I have identified some possible future work can be done in this area.

Collaborative explicit plasticity framework: a conceptual scheme for the generation of plastic and group-aware user interfaces

The advent of new advances in mobile computing has changed the manner we do our daily work, even enabling us to perform collaborative activities. However, current groupware approaches do not offer an integrating and efficient solution that jointly tackles the flexibility and heterogeneity inherent to mobility as well as the awareness aspects intrinsic to collaborative environments. Issues related to the diversity of contexts of use are collected under the term plasticity. A great amount of tools have emerged offering a solution to some of these issues, although always focused on individual scenarios. We are working on reusing and specializing some already existing plasticity tools to the groupware design. The aim is to offer the benefits from plasticity and awareness jointly, trying to reach a real collaboration and a deeper understanding of multi-environment groupware scenarios. In particular, this paper presents a conceptual framework aimed at being a reference for the generation of plastic User Interfaces for collaborative environments in a systematic and comprehensive way. Starting from a previous conceptual framework for individual environments, inspired on the model-based approach, we introduce specific components and considerations related to groupware.

IP Telephony in Third Generation Mobile Systems: Transport Layer Dimensioning

Tämä diplomityö käsittelee kolmannen sukupolven matkaviestinjärjestelmien kuljetuskerroksen mitoitusta. Nykyisten matkapuhelinverkkojen korvaajiksi suunnitellut kolmannen sukupolven matkaviestinjärjestelmät tulevat yhdistämään perinteisen puhelinviestinnän ja uudenlaiset datapalvelut. Uudet verkot tulevat perustumaan pakettivälitteiseen tiedonsiirtoon joka mahdollistaa molempien liikennetyyppien, puheen sekä datan, siirtämisen samassa verkossa. Tämän ratkaisun uskotaan tarjoavan paremmat mahdollisuudet uusien palvelujen luomiseen ja parantavan tiedonsiirtokapasiteettia. Siirtyminen pakettivälitteiseen tiedonsiirtoon aiheuttaa kuitenkin suuria muutoksia verkkoarkkitehtuurissa. Tässä diplomityössä tarkastellaan tulevien runkoverkkojen mitoitukseen liittyviä näkökohtia sekä muodostetaan alustavia kuljetuskerroksen mitoitusohjeita. Diplomityö on tehty osaksi diplomi-insinöörin tutkintoa Lappeenrannan teknillisessä korkeakoulussa. Työ on tehty Nokia Networksin palveluksessa Helsingissä, vuoden 2000 toisella puoliskolla.

Collecting Transaction Data in a Fourth Generation Mobile Network

Tässä diplomityössä on oletettu että neljännen sukupolven mobiiliverkko on saumaton yhdistelmä olemassa olevia toisen ja kolmannen sukupolven langattomia verkkoja sekä lyhyen kantaman WLAN- ja Bluetooth-radiotekniikoita. Näiden tekniikoiden on myös oletettu olevan niin yhteensopivia ettei käyttäjä havaitse saanti verkon muuttumista. Työ esittelee neljännen sukupolven mobiiliverkkoihin liittyvien tärkeimpien langattomien tekniikoiden arkkitehtuurin ja perustoiminta-periaatteet. Työ kuvaa eri tekniikoita ja käytäntöjä tiedon mittaamiseen ja keräämiseen. Saatuja transaktiomittauksia voidaan käyttää tarjottaessa erilaistettuja palvelutasoja sekä verkko- ja palvelukapasiteetin optimoimisessa. Lisäksi työssä esitellään Internet Business Information Manager joka on ohjelmistokehys hajautetun tiedon keräämiseen. Sen keräämää mittaustietoa voidaan käyttää palvelun tason seurannassa j a raportoinnissa sekä laskutuksessa. Työn käytännön osuudessa piti kehittää langattoman verkon liikennettä seuraava agentti joka tarkkailisi palvelun laatua. Agentti sijaitsisi matkapuhelimessa mitaten verkon liikennettä. Agenttia ei kuitenkaan voitu toteuttaa koska ohjelmistoympäristö todettiin vajaaksi. Joka tapauksessa työ osoitti että käyttäjän näkökulmasta tietoa kerääville agenteille on todellinen tarve.