Identification of in vitro phosphorylation sites in the growth cone protein SCG10. Effect Of phosphorylation site mutants on microtubule-destabilizing activity.

SCG10 is a neuron-specific, membrane-associated protein that is highly concentrated in growth cones of developing neurons. Previous studies have suggested that it is a regulator of microtubule dynamics and that it may influence microtubule polymerization in growth cones. Here, we demonstrate that in vivo, SCG10 exists in both phosphorylated and unphosphorylated forms. By two-dimensional gel electrophoresis, two phosphoisoforms were detected in neonatal rat brain. Using in vitro phosphorylated recombinant protein, four phosphorylation sites were identified in the SCG10 sequence. Ser-50 and Ser-97 were the target sites for protein kinase A, Ser-62 and Ser-73 for mitogen-activated protein kinase and Ser-73 for cyclin-dependent kinase. We also show that overexpression of SCG10 induces a disruption of the microtubule network in COS-7 cells. By expressing different phosphorylation site mutants, we have dissected the roles of the individual phosphorylation sites in regulating its microtubule-destabilizing activity. We show that nonphosphorylatable mutants have increased activity, whereas mutants in which phosphorylation is mimicked by serine-to-aspartate substitutions have decreased activity. These data suggest that the microtubule-destabilizing activity of SCG10 is regulated by phosphorylation, and that SCG10 may link signal transduction of growth or guidance cues involving serine/threonine protein kinases to alterations of microtubule dynamics in the growth cone.

Coordination on Networks: Does Topology Matter?

Effective coordination is key to many situations that affect the well-being of two or more humans. Social coordination can be studied in coordination games between individuals located on networks of contacts. We study the behavior of humans in the laboratory when they play the Stag Hunt game - a game that has a risky but socially efficient equilibrium and an inefficient but safe equilibrium. We contrast behavior on a cliquish network to behavior on a random network. The cliquish network is highly clustered and resembles more closely to actual social networks than the random network. In contrast to simulations, we find that human players dynamics do not converge to the efficient outcome more often in the cliquish network than in the random network. Subjects do not use pure myopic best-reply as an individual update rule. Numerical simulations agree with laboratory results once we implement the actual individual updating rule that human subjects use in our laboratory experiments.

Coordination and collaboration in European research towards healthy and safe nanomaterials

Nanotechnology is becoming part of our daily life in a wide range of products such as computers, bicycles, sunscreens or nanomedicines. While these applications already become reality, considerable work awaits scientists, engineers, and policy makers, who want such nanotechnological products to yield a maximum of benefit at a minimum of social, environmental, economic and (occupational) health cost. Considerable efforts for coordination and collaboration in research are needed if one wants to reach these goals in a reasonable time frame and an affordable price tag. This is recognized in Europe by the European Commission which funds not only research projects but also supports the coordination of research efforts. One of these coordination efforts is NanoImpactNet, a researcher-operated network, which started in 2008 promote scientific cross-talk across all disciplines on the health and environmental impact of nanomaterials. Stakeholders contribute to these activities, notably the definition of research and knowledge needs. Initial discussions in this domain focused on finding an agreement on common metrics, and which elements are needed for standardized approaches for hazard and exposure identification. There are many nanomaterial properties that may play a role. Hence, to gain the time needed to study this complex matter full of uncertainties, researchers and stakeholders unanimously called for simple, easy and fast risk assessment tools that can support decision making in this rapidly moving and growing domain. Today, several projects are starting or already running that will develop such assessment tools. At the same time, other projects investigate in depth which factors and material properties can lead to unwanted toxicity or exposure, what mechanisms are involved and how such responses can be predicted and modelled. A vision for the future is that once these factors, properties and mechanisms are understood, they can and will be accounted for in the development of new products and production processes following the idea of "Safety by Design". The promise of all these efforts is a future with nanomaterials where most of their risks are recognized and addressed before they even reach the market.

Quantitative modelling of transcription factor binding sites

Abstract One of the most important issues in molecular biology is to understand regulatory mechanisms that control gene expression. Gene expression is often regulated by proteins, called transcription factors which bind to short (5 to 20 base pairs),degenerate segments of DNA. Experimental efforts towards understanding the sequence specificity of transcription factors is laborious and expensive, but can be substantially accelerated with the use of computational predictions. This thesis describes the use of algorithms and resources for transcriptionfactor binding site analysis in addressing quantitative modelling, where probabilitic models are built to represent binding properties of a transcription factor and can be used to find new functional binding sites in genomes. Initially, an open-access database(HTPSELEX) was created, holding high quality binding sequences for two eukaryotic families of transcription factors namely CTF/NF1 and LEFT/TCF. The binding sequences were elucidated using a recently described experimental procedure called HTP-SELEX, that allows generation of large number (> 1000) of binding sites using mass sequencing technology. For each HTP-SELEX experiments we also provide accurate primary experimental information about the protein material used, details of the wet lab protocol, an archive of sequencing trace files, and assembled clone sequences of binding sequences. The database also offers reasonably large SELEX libraries obtained with conventional low-throughput protocols.The database is available at http://wwwisrec.isb-sib.ch/htpselex/ and and ftp://ftp.isrec.isb-sib.ch/pub/databases/htpselex. The Expectation-Maximisation(EM) algorithm is one the frequently used methods to estimate probabilistic models to represent the sequence specificity of transcription factors. We present computer simulations in order to estimate the precision of EM estimated models as a function of data set parameters(like length of initial sequences, number of initial sequences, percentage of nonbinding sequences). We observed a remarkable robustness of the EM algorithm with regard to length of training sequences and the degree of contamination. The HTPSELEX database and the benchmarked results of the EM algorithm formed part of the foundation for the subsequent project, where a statistical framework called hidden Markov model has been developed to represent sequence specificity of the transcription factors CTF/NF1 and LEF1/TCF using the HTP-SELEX experiment data. The hidden Markov model framework is capable of both predicting and classifying CTF/NF1 and LEF1/TCF binding sites. A covariance analysis of the binding sites revealed non-independent base preferences at different nucleotide positions, providing insight into the binding mechanism. We next tested the LEF1/TCF model by computing binding scores for a set of LEF1/TCF binding sequences for which relative affinities were determined experimentally using non-linear regression. The predicted and experimentally determined binding affinities were in good correlation.

Customer References as Marketing Practice in Company Web sites - Content and Discourse Analysis

Työn tarkoituksena oli tutkia sisältö- ja diskurssianalyysin avulla kuinka yritykset viestivät asiakasreferenssejä verkkosivuillaan. Työssä keskityttiin tutkimaan yritysten referenssikuvausten teemoja ja diskursseja, sekä sitä kuinka referenssisuhde rakentuu diskursiivisesti referenssikuvauksissa. Tutkimukseen valittiin kolme suomalaista ICT-alan yritystä: Nokia, TietoEnator ja F-Secure. Aineisto koostuu 140:stä yritysten WWW-sivuilta kerätystä referenssikuvauksesta. Sisältöanalyysin tuloksena havaittiin, että referenssikuvaukset keskittyvät kuvaamaan yksittäisiä tuote- tai projektitoimituksia referenssiasiakkaille kyseisten asiakassuhteiden valossa. Analyysin tuloksena tunnistettiin kolme diskurssia: hyötydiskurssi, sitoutumisen diskurssi sekä teknologisen eksperttiyden diskurssi. Diskurssit paljastavat referenssikuvausten retoriset keinot ja konstruoivat referenssisuhteen ja toimittajan subjektiposition eri näkökulmista. Pääpaino referenssikuvauksissa on toimittajan ratkaisun tuomissa hyödyissä. Diskurssit tuottavat referenssisuhteesta kuvan hyötyjä tuovana ja läheisenä asiakassuhteena, joka tarjoaa väylän ulkopuolisiin kyvykkyyksiin ja teknologioihin. Toimittaja esitetään referenssikuvauksissa diskurssista riippuen hyötyjen tuojana, luotettavana partnerina sekä kokeneena eksperttinä. Referenssiasiakas sen sijaan esitetään vain yhdestä näkökulmasta stereotyyppisesti tärkeänä ja tyytyväisenä asiakkaana.

Inter-limb coordination and energy cost in swimming.

OBJECTIVES: This study investigated the relationship between inter-arm coordination and the energy cost of locomotion in front crawl and breaststroke and explored swimmers' flexibility in adapting their motor organization away from their preferred movement pattern. DESIGN: Nine front-crawlers performed three 300-m in front crawl and 8 breaststrokers performed three 200-m in breaststroke at constant submaximal intensity and with 5-min rests. Each trial was performed randomly in a different coordination pattern: freely chosen, 'maximal glide' and 'minimal glide'. Two underwater cameras videotaped frontal and side views to analyze speed, stroke rate, stroke length and inter-limb coordination. METHODS: In front crawl, inter-arm coordination was quantified by the index of coordination (IdC) and the leg beat kicks were counted. In breaststroke, four time gaps quantified the arm to leg coordination (i.e., time between leg and arm propulsions; time between beginning, 90° flexion and end of arm and leg recoveries). The energy cost of locomotion was calculated from gas exchanges and blood lactate concentration. RESULTS: In both front crawl and breaststroke, the freely chosen coordination corresponded to glide pattern and showed the lowest energy cost (12.8 and 17.1Jkg(-1)m(-1), respectively). Both front-crawlers and breaststrokers were able to reach 'maximal glide' condition (respectively, +35% and +28%) but not 'minimal glide' condition for front crawl. CONCLUSIONS: The freely chosen pattern appeared more economic because more trained. When coordination was constrained, the swimmers had higher coordination flexibility in breaststroke than in front crawl, suggesting that breaststroke coordination was easier to regulate by changing glide time.

Detection of Body Dump Sites and Clandestine Burials: a GIS-Based Landscape Approach

Forensic archaeology has been of inestimable help in the location and excavation of clandestine burials (Hunter 1996a:16-17, 1996b; Killam 2004; Levine et al. 1984). Landscape archaeology techniques have been adapted and widely applied for such purposes. In this article, an adaptation of one of the most common applications of archaeological GIS, that is predictive site location, is applied to the detection of body dump sites and clandestine burial sites, bringing together in the process the fields of landscape archaeology, forensic sciences, and GIS.

Ohjelmistotuotteen hallintatietokannan suunnittelu ja toteutus

Ohjelmistotuotteen hallinta (SCM)on tärkeä osa ohjelmistoprojekteja. Se koostuu ohjelmistotuotteen hallinnan suunnittelusta, muutoksen hallinnasta, version hallinnasta, kääntämisestä, paketoinnista, kokoonpanon tilanteen seurannasta ja sen tarkistuksesta. Ohjelmistotuotteen hallintatietokanta (SCM DB) on tarkoitettu SCM:n liittyvändatan tallettamiseen yhteen paikkaan, jossa data on kaikkien löydettävissä. SCMDB on relaatiotietokanta ja WWW-käyttöliittymä sille. Tietokantaan talletetaan SCM - infrastruktuuri, SCM -resurssit, SCM -työskentelypaikat, integrointisuunnitteludata, paketoinnin raportit ja ohjeistukset, muutoksenhallintadata ja työkalujen hallintadata. Tietokannalla on monta käyttäjää. SCM managerit tallettavat tietokantaa yleiset tiedot, Integrointimanagerit tallettavat kantaan integrointisuunnitelmaa varten julkaisua koskevat tiedot. Paketointivastuulliset tallettavat kantaan paketointiraportit. Ohj elmistosuunnittelijat tekevät muutosvaatimuksia tietokantaan, jotka muutoksenhallintaelin käsittelee. He näkevät kannan kautta myös virheraportit. Työkalujen koordinointi tapahtuu myös kantaan talletettujen tietojen avulla. Lukemiseen tietokantaa voivat käyttää kaikki testauksesta suunnittelijoihin aikataulujen osalta. Tietokannasta voidaan lukea myös paketointityökalujen tallettamia tietoja ohjelmalohkoista eri pakettiversioissa. Paketointityökalut tai paketointivastuulliset saavat kannasta myös suoraan lähdetiedon paketointityökaluille.

Evaluation de la qualité des sites internet francophones traitant de la dépendance à l'alcool [Assessing the quality of French language web sites pertaining to alcohol dependency]

The objective of this article is to systematically assess the quality of web-based information in French language on the alcohol dependence. The authors analysed, using a standardised pro forma, the 20 most highly ranked pages identified by 3 common internet search engines using 2 keywords. Results show that a total of 45 sites were analysed. The authors conclude that the overall quality of the sites was relatively poor, especially for the description of possible treatments, however with a wide variability. Content quality was not correlated with other aspects of quality such as interactivity, aesthetic or accountability.

Genomic determinants of the efficiency of internal ribosomal entry sites of viral and cellular origin

Variation in cellular gene expression levels has been shown to be inherited. Expression is controlled at transcriptional and post-transcriptional levels. Internal ribosome entry sites (IRES) are used by viruses to bypass inhibition of cap-dependent translation, and by eukaryotic cells to control translation under conditions when protein synthesis is inhibited. We aimed at identifying genomic determinants of variability in IRES-mediated translation of viral [Encephalomyocarditis virus (EMCV)] and cellular IRES [X-linked inhibitor-of-apoptosis (XIAP) and c-myc]. Bicistronic lentiviral constructs expressing two fluorescent reporters were used to transduce laboratory and B lymphoblastoid cell lines [15 CEPH pedigrees (n = 205) and 50 unrelated individuals]. IRES efficiency varied according to cell type and among individuals. Control of IRES activity has a significant genetic component (h(2) of 0.47 and 0.36 for EMCV and XIAP, respectively). Quantitative linkage analysis identified a suggestive locus (LOD 2.35) on chromosome 18q21.2, and genome-wide association analysis revealed of a cluster of SNPs on chromosome 3, intronic to the FHIT gene, marginally associated (P = 5.9E-7) with XIAP IRES function. This study illustrates the in vitro generation of intermediate phenotypes by using cell lines for the evaluation of genetic determinants of control of elements such as IRES.

Coordination pattern adaptability: energy cost of degenerate behaviors.

This study investigated behavioral adaptability, which could be defined as a blend between stability and flexibility of the limbs movement and their inter-limb coordination, when individuals received informational constraints. Seven expert breaststroke swimmers performed three 200-m in breaststroke at constant submaximal intensity. Each trial was performed randomly in a different coordination pattern: 'freely-chosen', 'maximal glide' and 'minimal glide'. Two underwater and four aerial cameras enabled 3D movement analysis in order to assess elbow and knee angles, elbow-knee pair coordination, intra-cyclic velocity variations of the center of mass, stroke rate and stroke length and inter-limb coordination. The energy cost of locomotion was calculated from gas exchanges and blood lactate concentration. The results showed significantly higher glide, intra-cyclic velocity variations and energy cost under 'maximal glide' compared to 'freely-chosen' instructional conditions, as well as higher reorganization of limb movement and inter-limb coordination (p<0.05). In the 'minimal glide' condition, the swimmers did not show significantly shorter glide and lower energy cost, but they exhibited significantly lower deceleration of the center of mass, as well as modified limb movement and inter-limb coordination (p<0.05). These results highlight that a variety of structural adaptations can functionally satisfy the task-goal.