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Developing native applications for different platforms with different resolutions and screen sizes is both time consuming and costly. If developers were able to develop one web based application which can be used on multiple platforms, yet retain the same level of security as a native application, they would be able to reduce both development time and costs. In this thesis we will investigate the possibilities of achieving a level of security in a web-based application that can equal that of a native application, as well as how to develop an application that uses the Mina Vårdkontakter (My Healthcare Contacts) framework.

Einleitung: Somatische Mosaike des NLRP3 Gens wurden bei vielen „mutations-negativen“ CINCA Patienten detektiert. Wir versuchten solche low frequency NLRP3 Varianten bei „mutations-negativen“ Patienten nachzuweisen und zu quantifizieren, die an einem CINCA Syndrom, Muckle-Wells[for full text, please go to the a.m. URL], 42. Kongress der Deutschen Gesellschaft für Rheumatologie (DGRh); 28. Jahrestagung der Deutschen Gesellschaft für Orthopädische Rheumatologie (DGORh); 24. wissenschaftliche Jahrestagung der Gesellschaft für Kinder- und Jugendrheumatologie (GKJR)

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Transcriptome analysis of somatic stem cells and their progeny is fundamental to identify new factors controlling proliferation versus differentiation during tissue formation. Here, we generated a combinatorial, fluorescent reporter mouse line to isolate proliferating neural stem cells, differentiating progenitors and newborn neurons that coexist as intermingled cell populations during brain development. Transcriptome sequencing revealed numerous novel long non-coding (lnc)RNAs and uncharacterized protein-coding transcripts identifying the signature of neurogenic commitment. Importantly, most lncRNAs overlapped neurogenic genes and shared with them a nearly identical expression pattern suggesting that lncRNAs control corticogenesis by tuning the expression of nearby cell fate determinants. We assessed the power of our approach by manipulating lncRNAs and protein-coding transcripts with no function in corticogenesis reported to date. This led to several evident phenotypes in neurogenic commitment and neuronal survival, indicating that our study provides a remarkably high number of uncharacterized transcripts with hitherto unsuspected roles in brain development. Finally, we focussed on one lncRNA, Miat, whose manipulation was found to trigger pleiotropic effects on brain development and aberrant splicing of Wnt7b. Hence, our study suggests that lncRNA-mediated alternative splicing of cell fate determinants controls stem-cell commitment during neurogenesis.

Under slutet på 90-talet genomfördes mätningar av luften partikelhalter i sex svenska städer där syftet var att studera vilka halter som förekommer i gatu-, respektive taknivå och studera partiklarnas ursprung (Areskoug m. fl. 2001). Något förvånande visade det sig att den tydligt dominerande källan till partiklar i gatunivå under vinterhalvåret då halterna är som högst, inte är avgasrelaterade partiklar utan härrör från slitage av vägbana, däck och bromsar. Slitage av vägbanan orsakad av dubbdäcksanvändning bidrar mycket starkt till partikelhalterna, men även vintersandningen är en bidragande orsak. Den nyvunna kunskapen om slitagepartiklarnas betydelse för partikelhalterna kräver ökad kunskap om vilka faktorer som styr bildningen, om partiklarnas egenskaper, om deras eventuella betydelse för hälsoeffekter och om vilka åtgärder som kan bidra till att sänka bidraget. Projektet WearTox är ett av de första som Vägverket initierade för att fylla dessa kunskapsluckor. Projektet har utförts av VTI i samarbete med Lunds universitet och Linköpings universitet. Syftet med projektet har varit att studera den toxiska potentialen hos slitagepartiklar från beläggning och däck hos exponerade mänskliga luftvägsceller. slitagepartiklarnas fysikaliska, kemiska och morfologiska egenskaper. Under arbetets gång tillkom önskemål om karaktärisering av ytterligare kombinationer av däck vägbana och även från försök med sandad vägbana. Dessa tilläggsbeställningar inkluderade dock inte toxikologiska studier utan begränsade sig till karaktäriseringen av partiklarna.

Particles from road and tire wear have in recent years come to the fore as an important and relatively unexplored contribution to air particulate pollution. Mechanically generated wear particles are relatively coarse (>0.5 microns), but in controlled trials in VTI's road simulator it has been found that an ultra-fine fraction (< 100 nm) of particles is formed from tyre and pavement wear. This project was initiated to identify the source of these nanoparticles. The results show that the nanoparticles of the size 30-50 nm occur only from tests with studded tyres. One of the two tested Nordic unstudded winter tyres produced an even finer particle fraction (

Title Introduction Material and Methods Results Discussion Summary Zusammenfassung References Publications Supplementary Data Selbständigkeitserklärung, Functional genomics tries to understand the information encoded in the genome and its complex network organization. Thereby gene expression analysis and SNP genotyping are central elements of a functional genome analysis. Due to genome complexity, capable analysis methods with high-throughput capability are required in genomics. Limitations of available high-throughput methods such as microarrays are their relatively low sensitivity and reduced fexibility in assay design. This work aimed at developing a miniaturized high-density micro- well plate platform for liquid PCR-based assays to combine the high density of a miniaturized array format with the features of liquid PCR-based methods. With the polypropylene-based µPCR chip a cost-effective substrate for highly parallel PCR-based analysis was developed. The developed formats of the µPCR chip were verified to permit PCR-based assays in a volume range of 200 to 18 nl. Real-time PCR-based expression analysis was performed with the 5'-exonuclease assay (TaqMan). For reliable quantification down to 50 nl a workflow with advanced non-contact nanoliter liquid handling, efficient sealing, precise thermocycling and capable signal readout as well as data analysis procedures was developed. The platform demonstrated good reproducibility and high sensitivity down to single molecules in nanoliter volumes. The results of 200 nl assays in a study of tissue-specific gene expression in mouse were consistent with those obtained for 10 µl assays. Successful TaqMan-based SNP genotyping was performed down to 100 nl. For the analysis a robust allele-calling algorithm was developed. Due to the high sensitivity of the system, reliable genotyping was shown down to 5 initial target molecules of human genomic DNA as well as the equivalent amount of whole genome amplified DNA. In a validation study of 60 patients vs. 16 SNPs per chip, high concordance rates to the results obtained from conventional volumes were achieved. The newly developed µPCR chip platform proved to be a capable technology for miniaturized PCR-based analyzes in functional genomics. With the reduction of assay volumes down to few nanoliters the µPCR chip platform enables comprehensive large-scale studies in gene expression as well as SNP genotyping fine-mapping requiring only a moderate consumption of biological and financial resources., Die funktionelle Genomanalyse untersucht den Zusammenhang zwischen der im Genom codierten Information und deren AusprÄagung im Organismus. Dabei nehmen die Genexpressionsanalyse und die Genotypisierung von 'Single Nucleotide Polymorphismen'(SNP) als häufigste Form genetischer Variabilität eine zentrale Stellung ein. Durch die Komplexität des Genoms werden für die umfassende Analyse leistungsfähige Methoden benötigt, die hoch parallel arbeiten oder im Hochdurchsatz durchgeführt werden können. Vorhandene Methoden wie zum Beispiel die Microarray Technologie sind dabei oft durch geringe Sensitivität oder wenig Flexibilität in der Versuchsgestaltung begrenzt. Ziel dieser Arbeit war es, eine miniaturisierte Plattform für die Anwendung von flüssigen PCR- basierten Methoden zu entwickeln. Damit sollten die Vorteile einer sehr dichten Anordnung von Reaktionsgefäßen mit den Stärken PCR-basierter Methoden kombiniert werden. Mit dem aus Polypropylän hergestellten µPCR Chip wurde ein kosteneffektives Substrat für die hochgradig parallelisierte Analyse mit PCR- basierten Methoden entwickelt. Die verschieden Formate des µPCR Chip ermöglichen erfolgreiche PCR-Amplifikationen in Volumina zwischen 200 und 18 nl. Für die Genexpressionsanalyse wurde der 5'-Exonuklease Assay (TaqMan) als die geeigneteste Methode identifiziert, mit der die zuverlässige Quantifizierung bis zu einem Volumen von 50 nl erreicht wurde. Zur Durchführung der Analyse wurde leistungsfähiges Nanoliter-Handling, eine effektive Lösung zum Schließen der Reaktionsgefäße, Einrichtungen für die präzise thermische Prozessierung und eine leistungsfähige Detektion mit anschließender Datenalyse entwickelt. Neben einer guten Reproduzierbarkeit der Ergebnisse zeigte die Plattform eine hohe Sensitivität bis zu einzelnen Molekülen. In einer Validierungsstudie über gewebsspezifische Genexpression in Maus konnten mit 200 nl Reaktionen konsistente Ergebnisse zu den Ergebnissen in 10 µl gezeigt werden. Die erfolgreiche SNP Genotypisierung wurde ebenfalls mit dem 5'-Exonuklease Assay durchgeführt. Für die entwickelte Detektionsplattform wurde ein robuster Algorithmus zur Identifizierung der Allel-Information erarbeitet, der eine Reduktion des Reaktionsvolumens bis auf 100 nl ermöglichte. Durch die hohe Sensitivität der Plattform konnte bis zu einer Anfangsmenge von 5 DNA Molekülen genotypisiert werden. In einer Validierungsstudie mit 60 Patientenproben und 16 SNPs pro µPCR Chip wurde eine hohe Übereinstimmung zu den Ergebnissen aus konventionellen Reaktionsvolumina gezeigt. Mit der neu entwickelten µPCR Chip Plattform wurde eine leistungsfähige Technologie für miniaturisierte PCR-basierte Methoden in der Funktionellen Genomanalyse entwickelt. Durch die Reduktion des Reaktionsvolumens auf wenige Nanoliter werden großangelegte Genexpressionsstudien oder SNP Genotypisierungsstudien bei moderater Inanspruchnahme biologischer und finanzieller Ressourcen ermöglicht.

The high costs of maintenance related to pavement wear have caused many countries to prohibit the use of studded tyres. In Japan and Norway, regulations have also been based upon the health aspects of the road dust. In Japan studded tyres are prohibited and in Norway, restrictions have been placed on the use of studded tyres in some larger cities. In Sweden, wear particles from pavements, tyres, brakes and winter sanding/salting, have received a lot of attention the last few years. The reason for this is that measurements have shown that these particles are the main reason that the upcoming environmental quality standard for inhalable particles (PM10) will not be reached in many cities. The problem mainly occurs during dry episodes in winter and early spring and is mainly a result of studded tyre use and winter maintenance. A lot of new knowledge about wear particles is needed in order to develop cost-effective measures to deal with them. A great advantage is the possibility to study aerosols from each source both separately and in different combinations in a controlled environment. At VTI (Swedish National Road and Transport Research Institute) a road simulator (PVM), previously used for pavement and tyre wear studies, is used as a wear particle generator. The PVM can be equipped with different pavements and tyres and even friction materials such as sand can be applied. The PVM can be used at speeds up to 70 km/h and the pavement can be dry or wet. Also the temperature in the hall can be regulated below zero degrees centigrade. In this paper some results from the WearTox project will be presented. The production of PM10 from two pavements worn by studded tyres and friction tyres has been studied as well as the physical and chemical properties of the wear particles produced.

Airborne particles from railroad traffic have been identified as an air pollution problem mainly in tunnel environments. As a result of the introduction of the environmental quality standard for inhalable particles in Sweden 2005, the interest in railroad particle pollution has increased. The present report focuses on the variation of PM10 (inhalable particles) in some railroad environments under and above ground, the way these relate to traffic and on the particles' size distributions and elemental composition. The results show that particle concentrations in above ground railroad environments do not exceed the environmental quality standard during the campaigns. Diurnal mean values of PM10 range between 19 to 25 microg/m3. On the contrary, the diurnal mean concentrations on the platforms of the underground stations Arlanda Central and Arlanda South are far above the limit value (237 and 88 microg/m3 respectively) and clear diurnal and weekly patterns in PM10 concentration, co-fluctuating with traffic, can be identified. The particle mass size distribution has an obvious peak around 5-7 ?m at Arlanda C, and slightly smaller, 2-3 microm, at Arlanda S. The concentrations of both PM10 and ultrafine particles (< 0,1 microm) vary a lot depending on different trains. Especially the ultrafine particles seem to be emitted from certain trains, but it has not been possible to identify the source of these particles. The elemental composition of the particles in the tunnel environments was dominated by iron (84 % and 74 % respectively in Arlanda C and Arlanda S), but also other metals, like Cu, Zn, Cr, Ni and Sb (only at Arlanda C) have relatively high concentrations. The tunnel measurements also give some interesting results regarding possible measures against high particle concentrations. Washing of the tunnel walls and floor was carried out on two consecutive nights, but it had no noticeable results on particle concentrations. This implies that a dominant proportion of the particles is directly emitted rather than resuspended. On certain nights, the concentrations of all particle size fractions sank to very low levels, lasting till the morning traffic began. This implies effective self ventilation during these hours. Measurements in several different environments on Stockholm central station show that PM10 vary several tens of microg/m3. Activity, related to both traffic and people, increases particle concentration. The highest concentrations were measured inside the waiting hall. Nevertheless, the concentrations on the platforms were generally at least as high as in the busy street environment outside the station. During a railroad travel, the PM10 and PM2,5 concentrations were generally low, but increase at stops and, most prominently, on entering the Arlanda airport railroad tunnels. The PM2,5 proportion is considered high, contributing to approximately 50-80 % of PM10. The sources of railroad emitted PM10 are likely to be rails, wheels and brakes, but a source apportionment has not been accomplished in this project.

The aims of this project have been to describe PM10 from studded tyre wear and to study their inflammatory effects in human airway cells. The VTI circular road simulator has been used to generate "clean" wear particles from two different pavements; asphalt concrete (ABT) and stone mastic asphalt (ABS), with granite respectively quartzite as the main stone materials. The advantage of using the road simulator is that the contribution from other sources can be minimised. During the project time, the project was expanded also to study particle generation by non-studded winter tyres (friction tyres) and two kinds of winter sanding agents, namely washed crushed stone and unwashed natural sand in combination with both studded and friction tyres. Most of these parts of the project are presented in VTI report 520. The results show that pavement wear by studded tyres generates about 40-50 times as much PM10 as that by friction tyres, but also that the ABT pavement generates several times more PM10 than the ABS pavement. The size distribution within PM10 has a maximum around 3-4 ?m and more than 95 % of the mass is larger than 1 ?m. PM10 is totally dominated by fresh stone material. A fraction of very small particles, with a number concentration peak at around 30-40 nm, was also discovered. Their origin is unknown, but since the number distribution shifted depending on which tyre type was used, a possible origin is the tyres. In the cell studies the inflammation potential of the wear particles was compared with PM10 from Hornsgatan in Stockholm and PM10 from a Stockholm subway station. Later on in the project, diesel particles were made available for comparison. The cell study results show that PM10 from the ABT pavement is at least as inflammatory as diesel particles and more inflammatory than PM10 from the subway. The PM10 from Hornsgatan generally had the highest potential, but PM10 from the ABT pavement was often in parity. This despite the fact that the Hornsgatan PM10 were the only particle sample where endotoxin could be detected. Endotoxin causes an additive effect in studies of inflammation potential. PM10 from the ABS pavement generally had a lower potential than PM10 from the ABT pavement.