Role of ATH5 in the specification of retinal ganglion cells and expression and cell compartmentalization of EFEMP1, a protein associated with Malattia Leventinese

ABSTRACT : The development of the retina is a very complex process, occurring through the progressive restriction of cell fates, from pluripotent cell populations to complex tissues and organs. In all vertebrate species analyzed so far, retinal differentiation starts with the generation of retinal ganglion cells (RGC)s. One of the documented key essential events in the specification of RGCs is the expression of ATHS, an atonal homolog encoding a bHLH transcription factor. Despite the putative role of master regulator of RGC differentiation, the mechanism of integrating its functions into a coherent program underlying the production of this subclass of retinal neurons has not yet been elucidated. By using chromatin immunoprecipitation combined with microarray (ChIP-on-chip) we have screened for ATH5 direct targets in the developing chick retina at two consecutive periods: E3.5 (stage HH22) and E6 (stage HH30), covering the stages of progenitor proliferation, neuroepithelium patterning, RGC specification, cell cycle exit and early neuronal differentiation. In parallel, complementary analysis with Affymetrix expression microarrays was conducted. We compared RGCs versus retina to see if the targets correspond to genes preferentially expressed in RGCs. We also precociously overexpressed ATH5 in the retina of individual embryo, and contralateral retina vas used as a control. Our integrated approach allowed us to establish a compendium of ATH5-targets and enabled us to position ATH5 in the transcription network underlying neurogenesis in the retina. Malattia Leventinese (ML) is an autosomal, dominant retinal dystrophy characterized by extracellular, amorphous deposits known as drusen, between the retinal pigment epithelium (RPE) and Bruch's membrane. On the genetic level, it has been associated with a single missense mutation (R345W) in a widely expressed gene with unknown function called EFEMP1. We determined expression patterns of the EFEMP1 gene in normal and ML human retinas. Our data shown that the upregulation of EFEMP1 is not specific to ML eye, except for the region of the ciliary body. We also analyzed the cell compartmentalization of different versions of the protein (both wild type and mutant). Our studies indicate that both abnormal expression of the EFEMP1 gene and mutation and accumulation of EFEMP 1 protein (inside or outside the cells) might contribute to the ML pathology. Résumé : 1er partie : L'ontogenèse de la rétine est un processus complexe au cours duquel des cellules progénitrices sont engagée, par vagues successives, dans des lignées où elles vont d'abord être déterminées puis vont se différencier pour finalement construire un tissu rétinien composé de cinq classes de neurones (les photorécepteurs, les cellules horizontales, bipolaires, amacrines et ganglionnaires) et d'une seule de cellules gliales (les cellules de Muller). Chez tous les vertébrés, la neurogenèse rétinienne est d'abord marquée par la production des cellules ganglionnaires (RGCs). La production de cette classe de neurone est liée à l'expression du gène ATH5 qui est un homologue du gène atonal chez la Drosophile et qui code pour un facteur de transcription de la famille des protéines basic Helix-Loop-Helix (bHLH). Malgré le rôle central que joue ATH5 dans la production des RGCs, le mécanisme qui intègre la fonction de cette protéine dans le programme de détermination neuronale et ceci en relation avec le développement de la rétine n'est pas encore élucidé. Grâce à une technologie qui permet de combiner la sélection de fragments de chromatine liant ATH5 et la recherche de séquences grâce à des puces d'ADN non-codants (ChIP-on-chip), nous avons recherché des cibles potentielles de la protéine ATH5 dans la rétine en développement. Nous avons conduit cette recherche à deux stades de développement de manière à englober la phase de prolifération cellulaire, la détermination des RGCs, la sortie du cycle cellulaire ainsi que les premières étapes de la différentiation de ces neurones. Des expériences complémentaires nous ont permis de définir les patrons d'expression des gènes sélectionnés ainsi que l'activité promotrice des éléments de régulation identifiés lors de notre criblage. Ces approches expérimentales diverses et complémentaires nous ont permis de répertorier des gènes cibles de la protéine ATH5 et d'établir ainsi des liens fonctionnels entre des voies métaboliques dont nous ne soupçonnions pas jusqu'alors qu'elles puissent être associées à la production d'une classe de neurones centraux. 2ème partie : Malattia Leventinese (ML) est une maladie génétique qui engendre une dystrophie de la rétine. Elle se caractérise par l'accumulation de dépôt amorphe entre l'épithélium pigmentaire et la membrane de Bruch et connu sous le nom de drusen. Cette maladie est liée à une simple mutation non-sens (R345W) dans un gène dénommé EFEMP1 qui est exprimé dans de nombreux tissus mais dont la fonction reste mal définie. Une étude détaillée de l'expression de ce gène dans des rétines humaines a révélé une expression à un niveau élevé du gène EFEMP1 dans divers tissus de l'oeil ML mais également dans des yeux contrôles. Alors que l'accumulation d'ARN messager EFEMP1 dans les cellules de l'épithélium pigmentaire n'est pas spécifique à ML, l'expression de ce gène dans le corps cilié n'a été observée que dans l'oeil ML. Nous avons également comparé la sécrétion de la protéine sauvage avec celle porteuse de la mutation. En résumé, notre étude révèle que le niveau élevé d'expression du gène EFEMP1 ainsi que l'accumulation de la protéine dans certains compartiments cellulaires pourraient contribuer au développement de pathologies rétiniennes liées à ML.

A TARBP2-dependent miRNA expression profile underlies cancer stem cell properties and provides candidate therapeutic reagents in Ewing sarcoma.

We have recently demonstrated that human pediatric mesenchymal stem cells can be reprogrammed toward a Ewing sarcoma family tumor (ESFT) cancer stem cell (CSC) phenotype by mechanisms that implicate microRNAs (miRNAs). Here, we show that the miRNA profile of ESFT CSCs is shared by embryonic stem cells and CSCs from divergent tumor types. We also provide evidence that the miRNA profile of ESFT CSCs is the result of reversible disruption of TARBP2-dependent miRNA maturation. Restoration of TARBP2 activity and systemic delivery of synthetic forms of either of two of its targets, miRNA-143 or miRNA-145, inhibited ESFT CSC clonogenicity and tumor growth in vivo. Our observations suggest that CSC self-renewal and tumor maintenance may depend on deregulation of TARBP2-dependent miRNA expression.

Assessing an intuitive condition for stability under a range of traffic conditions via a generalised Lu-Kumar network

We argue the importance both of developing simple sufficientconditions for the stability of general multiclass queueing networks and also of assessing such conditions under a range of assumptions on the weight of the traffic flowing between service stations. To achieve the former, we review a peak-rate stability condition and extend its range of application and for the latter, we introduce a generalisation of the Lu-Kumar network on which the stability condition may be tested for a range of traffic configurations. The peak-rate condition is close to exact when the between-station traffic is light, but degrades as this traffic increases.

CD93 is required for maintenance of antibody secretion and persistence of plasma cells in the bone marrow niche.

Plasma cells represent the end stage of B-cell development and play a key role in providing an efficient antibody response, but they are also involved in numerous pathologies. Here we show that CD93, a receptor expressed during early B-cell development, is reinduced during plasma-cell differentiation. High CD93/CD138 expression was restricted to antibody-secreting cells both in T-dependent and T-independent responses as naive, memory, and germinal-center B cells remained CD93-negative. CD93 was expressed on (pre)plasmablasts/plasma cells, including long-lived plasma cells that showed decreased cell cycle activity, high levels of isotype-switched Ig secretion, and modification of the transcriptional network. T-independent and T-dependent stimuli led to re-expression of CD93 via 2 pathways, either before or after CD138 or Blimp-1 expression. Strikingly, while humoral immune responses initially proceeded normally, CD93-deficient mice were unable to maintain antibody secretion and bone-marrow plasma-cell numbers, demonstrating that CD93 is important for the maintenance of plasma cells in bone marrow niches.

Voltage fluctuations in IC power supply distribution

The supply voltage decrease and powerconsumption increase of modern ICs made the requirements for low voltage fluctuation caused by packaging and on-chip parasitic impedances more difficult to achieve. Most of the research works on the area assume that all the nodes of the chip are fed at thesame voltage, in such a way that the main cause of disturbance or fluctuation is the parasitic impedance of packaging. In the paper an approach to analyze the effect of high and fast current demands on the on-chip power supply network. First an approach to model the entire network by considering a homogeneous conductive foil is presented. The modification of the timing parameters of flipflops caused by spatial voltage drops through the IC surface are also investigated.

Kaukolämmöntuotannon ja verkon käytön optimointi uudessa ajotilanteessa

Diplomityössä kehitetään Savonlinnan kaupungin kaukolämmöntuotantoa ja kaukolämpöverkon käyttöä muuttuneessa ajotilanteessa. Muuttunut tilanne syntyy, kun Savonlinnaan rakennetaan uusi lämmitysvoimalaitos. Kaukolämpöverkkoa yhdistetään samanaikaisesti viidestä erillisestä verkosta yhdeksi kokonaiseksi verkoksi. Kaukolämmöntuotannon ja verkon käytön optimointi suoritettiin Process Vision Oy: n kehittämällä kaukolämpöverkon laskentaohjelmalla. Optimoinnissa pyrittiin saamaan mahdollisimman aikaisin taloudellisin laitos eli uusi hakelaitos täyteen tehoon ja tarvittava lisäteho otettiin öljylämpökeskuksista. Hakelaitoksen käytettävyyttä lisättiin rakentamalla kaukolämpöverkkoon välipumppaamo ja kaukolämpöveden apujäähdytin. Hakelaitosta voidaan käyttää 0°C ulkolämpötilaan asti, mutta kun käytetään apuna välipumppausta voidaan pumppauksellisesti pelkästään hakelaitokselta syöttää tehoa aina 14 °C lämpötilaan asti. Välipumppauksen avulla vuosittain vähennetään öljyn ja lisätään hakkeen polttoa n. 10,4 GWh. Nykyisillä öljyn ja hakkeen hinnoilla säästö vuodessa on n. 887000 mk. Välipumppauksella vähennetään lisäksi kaukolämpöverkon pumppauskustannuksia.

Regional Data Transmission Network

The globalization and development of an information society promptly change shape of the modern world. Cities and especially megacities including Saint-Petersburg are in the center of occuring changes. As a result of these changes the economic activities connected to reception and processing of the information now play very important role in economy of megacities what allows to characterize them as "information". Despite of wide experience in decision of information questions Russia, and in particular Saint-Petersburg, lag behind in development of information systems from the advanced European countries. The given master's thesis is devoted to development of an information system (data transmission network) on the basis of wireless technology in territory of Saint-Petersburg region within the framework of FTOP "Electronic Russia" and RTOP "Electronic Saint-Petersburg" programs. Logically the master's thesis can be divided into 3 parts: 1. The problems, purposes, expected results, terms and implementation of the "Electronic Russia" program. 2. Discussion about wireless data transmission networks (description of technology, substantiation of choice, description of signal's transmission techniques and types of network topology). 3. Fulfillment of the network (organization of central network node, regional centers, access lines, description of used equipment, network's capabilities), financial provision of the project, possible network management models.

Creating a research network for a successful e-portfolio design and implementation

The presence of e-portfolios in educational centres, companies and administrations has emergedstrongly during the last years by creating very different practices coming from different objectives and purposes. This situation has led researchers and practitioners to design and implement e-portfolios with little reference to previous knowledge of them; consequently, developments are disparate with many of the processes and dimensions used both in development and use being unnecessary complex. In order to minimize the inconveniences, unify these developmental processes and improve the resultsof implementation and use of e-portfolios, it seemed necessary to create a network of researchers, teachers and trainers coming from different universities and institutions of different kinds who are interested in the investigation and the practice of e-portfolios in Spain. Therefore, The Network on e-portfoliowas created in 2006, funded by the Spanish Ministry of Education and led by the UniversitatOberta de Catalunya. Besides the goals associatedwith the creation of this network and which wewanted to share with other European researchers and experts of other continents, we will also present in this paper some data concerned with the first study carried out on the use of e-portfolios in our country that shows where we are and which trends are the most important for the near future.

Phase voltage control and filtering in a converter-fed single-phase customer-end system of the LVDC distribution network

In recent years, the network vulnerability to natural hazards has been noticed. Moreover, operating on the limits of the network transmission capabilities have resulted in major outages during the past decade. One of the reasons for operating on these limits is that the network has become outdated. Therefore, new technical solutions are studied that could provide more reliable and more energy efficient power distributionand also a better profitability for the network owner. It is the development and price of power electronics that have made the DC distribution an attractive alternative again. In this doctoral thesis, one type of a low-voltage DC distribution system is investigated. Morespecifically, it is studied which current technological solutions, used at the customer-end, could provide better power quality for the customer when compared with the current system. To study the effect of a DC network on the customer-end power quality, a bipolar DC network model is derived. The model can also be used to identify the supply parameters when the V/kW ratio is approximately known. Although the model provides knowledge of the average behavior, it is shown that the instantaneous DC voltage ripple should be limited. The guidelines to choose an appropriate capacitance value for the capacitor located at the input DC terminals of the customer-end are given. Also the structure of the customer-end is considered. A comparison between the most common solutions is made based on their cost, energy efficiency, and reliability. In the comparison, special attention is paid to the passive filtering solutions since the filter is considered a crucial element when the lifetime expenses are determined. It is found out that the filter topology most commonly used today, namely the LC filter, does not provide economical advantage over the hybrid filter structure. Finally, some of the typical control system solutions are introduced and their shortcomings are presented. As a solution to the customer-end voltage regulation problem, an observer-based control scheme is proposed. It is shown how different control system structures affect the performance. The performance meeting the requirements is achieved by using only one output measurement, when operating in a rigid network. Similar performance can be achieved in a weak grid by DC voltage measurement. An additional improvement can be achieved when an adaptive gain scheduling-based control is introduced. As a conclusion, the final power quality is determined by a sum of various factors, and the thesis provides the guidelines for designing the system that improves the power quality experienced by the customer.

Thermal-Aware Networked Many-Core Systems

Advancements in IC processing technology has led to the innovation and growth happening in the consumer electronics sector and the evolution of the IT infrastructure supporting this exponential growth. One of the most difficult obstacles to this growth is the removal of large amount of heatgenerated by the processing and communicating nodes on the system. The scaling down of technology and the increase in power density is posing a direct and consequential effect on the rise in temperature. This has resulted in the increase in cooling budgets, and affects both the life-time reliability and performance of the system. Hence, reducing on-chip temperatures has become a major design concern for modern microprocessors. This dissertation addresses the thermal challenges at different levels for both 2D planer and 3D stacked systems. It proposes a self-timed thermal monitoring strategy based on the liberal use of on-chip thermal sensors. This makes use of noise variation tolerant and leakage current based thermal sensing for monitoring purposes. In order to study thermal management issues from early design stages, accurate thermal modeling and analysis at design time is essential. In this regard, spatial temperature profile of the global Cu nanowire for on-chip interconnects has been analyzed. It presents a 3D thermal model of a multicore system in order to investigate the effects of hotspots and the placement of silicon die layers, on the thermal performance of a modern ip-chip package. For a 3D stacked system, the primary design goal is to maximise the performance within the given power and thermal envelopes. Hence, a thermally efficient routing strategy for 3D NoC-Bus hybrid architectures has been proposed to mitigate on-chip temperatures by herding most of the switching activity to the die which is closer to heat sink. Finally, an exploration of various thermal-aware placement approaches for both the 2D and 3D stacked systems has been presented. Various thermal models have been developed and thermal control metrics have been extracted. An efficient thermal-aware application mapping algorithm for a 2D NoC has been presented. It has been shown that the proposed mapping algorithm reduces the effective area reeling under high temperatures when compared to the state of the art.

Effects of lipopolysaccharide on low- and high-density cultured rabbit vascular smooth muscle cells: differential modulation of nitric oxide release, ERK1/ERK2 MAP kinase activity, protein tyrosine phosphatase activity, and DNA synthesis

Previous studies have shown that exogenously generated nitric oxide (NO) inhibits smooth muscle cell proliferation. In the present study, we stimulated rabbit vascular smooth muscle cells (RVSMC) with E. coli lipopolysaccharide (LPS), a known inducer of NO synthase transcription, and established a connection between endogenous NO, phosphorylation/dephosphorylation-mediated signaling pathways, and DNA synthesis. Non-confluent RVSMC were cultured with 0, 5, 10, or 100 ng/ml of the endotoxin. NO release was increased by 86.6% (maximum effect) in low-density cell cultures stimulated with 10 ng/ml LPS as compared to non-stimulated controls. Conversely, LPS (5 to 100 ng/ml) did not lead to enhanced NO production in multilayered (high density) RVSMC. DNA synthesis measured by thymidine incorporation showed that LPS was mitogenic only to non-confluent RVSMC; furthermore, the effect was prevented statistically by aminoguanidine (AG), a potent inhibitor of the inducible NO synthase, and oxyhemoglobin, an NO scavenger. Finally, there was a cell density-dependent LPS effect on protein tyrosine phosphatase (PTP) and ERK1/ERK2 mitogen-activated protein (MAP) kinase activities. Short-term transient stimulation of ERK1/ERK2 MAP kinases was maximal at 12 min in non-confluent RVSMC and was prevented by preincubation with AG, whereas PTP activities were inhibited in these cells after 24-h LPS stimulation. Conversely, no significant LPS-mediated changes in kinase or phosphatase activities were observed in high-density cells. LPS-induced NO generation by RVSMC may switch on a cell density-dependent proliferative signaling cascade, which involves the participation of PTP and the ERK1/ERK2 MAP kinases.

Nettikoulussa – Opettajien käsityksiä nettikasvatuksesta ja nuorten nettikulttuurista

Tutkimuksessa kuvataan opettajien käsityksiä nettikasvatuksesta ja nuorten nettikulttuurista. Tehtävänä on kuvata kuuden opettajan näkemyksiä nuorten nettimaailmasta sekä oppilaan, että kasvattajan näkökulmasta. Tutkimus on lähtökohdiltaan kvalitatiivinen. Tutkimuksen metodologisena viitekehyksenä käytetään fenomenografista lähestymistapaa. Tutkimuksen tulosten mukaan opettajien käsitykset nuorten nettikulttuurista olivat hyvin moninaiset. Opettajien käsityksiä nuorten nettikulttuurista kuvasti vahva huoli nuorten arvoista ja suhteestaan mediakriittisyyteen. Nettikulttuuriin liittyen huolenaiheena olivat myös nuorten nettiriippuvuus, sekä huoli liikunnan vähentymisestä. Nämä aiheet heijastuivat vahvasti opettajien omiin nuoruudenajan kokemuksiin nuorisokulttuurista. Toisaalta internet ja sosiaalinen media nähtiin mahdollistavana, yhdistävänä tekijänä, joka heijastui koulun arkeen nuorten avoimuutena ja kokemusten jakamisena. Opettajien käsityksissä heijastui myös kulttuurin kokonaisvaltaisuus; miten Internet ja sosiaalinen media syö kulttuurin itseensä, kun kaikki toiminta siirtyy verkkoon. Nettikasvatuksen opettajat kokivat olevan hyvin harkinnanvaraista ja riippuvan omasta kiinnostuksesta aihetta kohtaan. Nettikasvattajina opettajat kokivat tärkeäksi mediakasvatuksen ja kriittisen tiedon etsinnän taidot. Vastuu ja sen jakaminen olivat merkittävä tekijä opettajien puhuessa nettikasvatuksestaan ja roolistaan siinä. Opettajat eivät halunneet mennä kodin tehtävien edelle, mutta rajan määrittäminen oli haasteellista. Vastavuoroisuus nettikasvatuksessa ja teknologisessa koulumaail-massa nähtiin hyvin merkittävä.

Identification in silico d’éléments de réponse de récepteurs nucléaires impliqués dans le cancer du sein

La croissance de deux tiers des tumeurs mammaires dépend des œstrogènes. Le réseau de gènes responsable de propager les signaux prolifératifs des œstrogènes est encore mal connu. Des micropuces d’ADN de cellules de carcinome mammaire MCF7 traitées à l’œstradiol (E2) avec ou sans l’inhibiteur de synthèse protéique cycloheximide (CHX) ont permis d’identifier de nombreux gènes cibles primaires et secondaires. La séquence des promoteurs des gènes cibles a été criblée à l’aide d’une banque de 300 matrices modélisant les sites reconnus par divers facteurs de transcription. Les éléments de réponse aux œstrogènes (ERE) sont enrichis dans les promoteurs des gènes primaires. Les sites E2F sont enrichis dans les promoteurs des gènes cible secondaires. Un enrichissement similaire a été observé avec les régions liées par ERα et E2F1 en ChIP-on-chip pour chacune des catégories de gènes. La croissance des cellules de carcinome mammaire est inhibée par des traitements à l’acide rétinoïque (RA). L’analyse de micropuces d’ADN de MCF7 traitées avec RA a permis d’identifier de nombreux gènes cibles potentiels. Un enrichissement d’éléments de réponse à l’acide rétinoïque (RARE) est observable dans les promoteurs de ces gènes après avoir exclus les RARE se trouvant à l’intérieur d’éléments transposables. Des RARE présents dans des éléments transposables spécifiques aux primates sont aussi fixés in vivo dans les promoteurs de cibles connues de RA : BTG2, CASP9 et GPRC5A. Certains gènes cibles de RA dans les MCF7 sont aussi des cibles de E2, suggérant que le contrôle que ces molécules exercent sur la prolifération est en partie attribuable à des effets opposés sur un ensemble commun de gènes.

Influence of occupation number of single particle levels on K - K charge transfer in collisions of 90 keV - Ne{^9+} on Ne

The influence of the occupation of the single particle levels on the impact parameter dependent K - K charge transfer occuring in collisions of 90 keV Ne{^9+} on Ne was studied using coupled channel calculations. The energy eigenvalues and matrixelements for the single particle levels were taken from ab initio self consistent MO-LCAO-DIRAC-FOCK-SLATER calculations with occupation numbers corresponding to the single particle amplitudes given by the coupled channel calculations.

Conditions for Viewpoint Dependent Face Recognition

Poggio and Vetter (1992) showed that learning one view of a bilaterally symmetric object could be sufficient for its recognition, if this view allows the computation of a symmetric, "virtual," view. Faces are roughly bilaterally symmetric objects. Learning a side-view--which always has a symmetric view--should allow for better generalization performances than learning the frontal view. Two psychophysical experiments tested these predictions. Stimuli were views of shaded 3D models of laser-scanned faces. The first experiment tested whether a particular view of a face was canonical. The second experiment tested which single views of a face give rise to best generalization performances. The results were compatible with the symmetry hypothesis: Learning a side view allowed better generalization performances than learning the frontal view.