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Left-right asymmetry is an important organizing feature of the healthy brain that may be altered in schizophrenia, but most studies have used relatively small samples and heterogeneous approaches, resulting in equivocal findings. We carried out the largest case-control study of structural brain asymmetries in schizophrenia, with MRI data from 5,080 affected individuals and 6,015 controls across 46 datasets, using a single image analysis protocol. Asymmetry indexes were calculated for global and regional cortical thickness, surface area, and subcortical volume measures. Differences of asymmetry were calculated between affected individuals and controls per dataset, and effect sizes were meta-analyzed across datasets. Small average case-control differences were observed for thickness asymmetries of the rostral anterior cingulate and the middle temporal gyrus, both driven by thinner left-hemispheric cortices in schizophrenia. Analyses of these asymmetries with respect to the use of antipsychotic medication and other clinical variables did not show any significant associations. Assessment of age- and sex-specific effects revealed a stronger average leftward asymmetry of pallidum volume between older cases and controls. Case-control differences in a multivariate context were assessed in a subset of the data (N = 2,029), which revealed that 7% of the variance across all structural asymmetries was explained by case-control status. Subtle case-control differences of brain macrostructural asymmetry may reflect differences at the molecular, cytoarchitectonic, or circuit levels that have functional relevance for the disorder. Reduced left middle temporal cortical thickness is consistent with altered left-hemisphere language network organization in schizophrenia.

This project contains a preregistration of secondary data analysis for a research project on the complex links between the genetics influence on the wellbeing spectrum, brain structure, and risk for psychopathology in children.

Background Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer with a dismal prognosis. Although combined treatment with gemcitabine and albumin-bound paclitaxel has improved the prognosis of PDAC, both intrinsic and acquired chemoresistance remain as severe hurtles towards improved prognosis. Thus, new therapeutic targets and innovative strategies are urgently needed. Methods In this study, we used the KPC mouse model-derived PDAC cell line TB32047 to perform kinome-wide CRISPR-Cas9 loss-of-function screening. Next-generation sequencing and MAGeCK-VISPR analysis were performed to identify candidate genes. We then conducted cell viability, clonogenic, and apoptosis assays and evaluated the synergistic therapeutic effects of cyclin-dependent kinase 7 (CDK7) depletion or inhibition with gemcitabine (GEM) and paclitaxel (PTX) in a murine orthotopic pancreatic cancer model. For mechanistic studies, we performed genome enrichment analysis (GSEA) and Western blotting to identify and verify the pathways that render PDAC sensitive to GEM/PTX therapy. Results We identified several cell cycle checkpoint kinases and DNA damage-related kinases as targets for overcoming chemoresistance. Among them, CDK7 ranked highly in both screenings. We demonstrated that both gene knockout and pharmacological inhibition of CDK7 by THZ1 result in cell cycle arrest, apoptosis induction, and DNA damage at least predominantly through the STAT3-MCL1-CHK1 axis. Furthermore, THZ1 synergized with GEM and PTX in vitro and in vivo, resulting in enhanced antitumor effects. Conclusions Our findings support the application of CRISPR-Cas9 screening in identifying novel therapeutic targets and suggest new strategies for overcoming chemoresistance in pancreatic cancer.

Historien og de kollektive minnene om andre verdenskrig har mange forskjellige vinkler og tilnærminger, og museet har en helt sentral plass som formidler av disse. Røde Kors Krigsminnemuseum i Narvik åpnet i mars 1964. Gjennom en utvikling på over 50 år, har en institusjon for historisk formidling vært et bevaringssted for krigshistoriske gjenstander som har blitt presentert for et bredt utvalg av publikum. Fra 2016 gikk museet inn i en ny fase som innebar overgangen og utviklingen til det som nå er kjent som Narviksenteret. Museet som institusjon blir i seg selv et historisk åsted, med fortellinger som bidrar til å utvikle et minne om krigen. Denne studien ser på Røde Kors Krigsminnemuseum og Narviksenteret som forvalter og formidler av en fortelling om krigen i Narvik under andre verdenskrig. Ved å undersøke hvordan andre verdenskrig i Norge og Narvik har blitt fremstilt gjennom etterkrigstiden, kaster dette prosjektet lys på de mange prosessene og fasene Røde Kors Krigsminnemuseum og Narviksenteret har gjennomgått, og hvordan det kollektive minnet kan ha blitt påvirket av dette. Det kollektive minnet og fortellingen om andre verdenskrig har vært viktig for at nye generasjoner kan forstå å lære av tidligere hendelser. Et sentralt funn i denne undersøkelsen er at det har vært en klar utvikling bort fra det patriotiske narrativet som ble fremstilte i de tidlige fasene etter krigen. Det har gått fra å sette søkelys på motgang og seier hos Røde Kors Krigsminnemuseum, til mer generelle moralske spørsmål hos Narviksenteret.

Additional file 1: Fig. S1. FACS-plot of D. melanogaster eye-antennal disc cells after live/dead cell staining. (A) Counterstaining of Propidium Iodide to label dead cells (y-axis, Q1) and Calcein violet to label live cells (x-axis, Q4). Double positive signals might indicate dying cells or incompletely separated cells (Q2). This method allows removing debris (Q3) efficiently. (B) The cell population P4 (i.e. 16,208 living cells) were isolated and used for the scRNAseq run using 10x Genomics. Fig. S2. Quality and quantity of cDNA after reverse transcription of mRNA fraction (polyA-based enrichment) and full-length cDNA amplification from cell lysate of 30 cells sorted from cell suspension of eye-antennal discs run on Fragment Analyzer (Agilent). Size distribution of all fragments shows little impact on degradation (almost no cDNA detectable below 400 bp). Fig. S3. Contribution of mitochondrial gene expression to scRNAseq and snRNAseq datasets. (A) Total amount of genes (features) over percentage of mitochondrial reads, per cell each. The dashed line indicates a threshold of 10% of reads attributed to mitochondrial genes. In scRNAseq data, approximately 14% of cells show a high (>10%) proportion of mitochondrial gene reads on the total number of reads. (B) Total amount of genes (features) over Percentage of mitochondrial reads, per cell each in snRNAseq data. The dashed line indicates a threshold of 10% of reads attributed to mitochondrial genes. In most nuclei, only a low percentage of reads is attributed to mitochondrial genes. Fig. S4. Clustering and cluster annotation for scRNAseq data. (A) The heatmap shows the score for each potential cell type (Y-axis) in each cluster (X-axis). The cell types are annotated based on the highest scoring identity in the heatmap. The clusters are grouped based on their transcriptional similarity to each other. For clusters which express an equal number of marker genes for two different identities both identities were assigned (e.g. cluster 3:Antenna|EAB). Clusters with unresolved identities (i.e. more than two equal assignments) are called “Other”. The colors of the cluster names correspond to the colors in UMAP in (B). The marker score is calculated using a matrix of published marker genes (see Additional file Table S2). (B) UMAP of scRNAseq data. The clusters were annotated based on the heatmap in (A). This UMAP is identical to the UMAP with combined cluster annotation shown in Fig. 3A. Note that the color code is not comparable to the one used in Additional file Figs. S7 and S10. Fig. S5. Fluorescence intensity curves from Bioanalyzer for fresh- and cryopreserved nuclei obtained by different nuclei extraction protocols. (A) The RNA was extracted directly from a fresh sample (36 eye-antennal discs), which was dissociated using the 10x Genomics protocol with 0.1% IGEPAL as a detergent. (B) RNA isolated from a cryopreserved sample, which was dissociated using a protocol based on Triton X-100 as a detergent and a variety of RNAse inhibitors [1]. Note that the sample was thawed for 3.5h before being frozen again. Both curves are close to the expectation of RNA isolated from D. melanogaster [2]. Fig. S6. BioAnalyzer results comparing different nuclei isolation protocols for frozen samples. Samples 1, 2, 4 and 5 were dissociated using the protocol based on Triton X-100 as a detergent and a variety of RNAse inhibitors [1]. Samples 1 and 2 were dissociated by pipetting up and down and samples 4 and 5 were dissociated using a Dounce homogenizer. Samples 7 and 8 were dissociated using the protocol “10x Genomics® Isolation of Nuclei for Single Cell RNA Sequencing” [3]. Samples 7 and 8 were dissociated using only citric acid buffer and samples 10 und 11 were dissociated using only a detergent. Note that for D. melanogaster, intense bands are expected at about 40s and weaker bands at 25s and 45-50s [2]. Each run was repeated once. Fig. S7. Clustering and cluster annotation for snRNAseq data. (A) The heatmap shows the score for each potential cell type (Y-axis) in each cluster (X-axis). The cell types are annotated based on the highest scoring identity in the heatmap. The clusters are grouped based on their transcriptional similarity to each other. For clusters which express an equal number of marker genes for two different identities both identities were assigned (e.g. cluster 15:MFurrow|SMW). Clusters with unresolved identities (i.e. more than two equal assignments) are called “Other”. The colors of the cluster names correspond to the colors in UMAP in (B). The marker score is calculated using a matrix of published marker genes (see Additional file Table S2). (B) UMAP of snRNAseq data. The clusters were annotated based on the heatmap in (A). This UMAP is identical to the UMAP with combined cluster annotation shown in Fig. 3C. Note that the color code is not comparable to the one used in Additional file Figs. S4 and S10. Fig. S8. Gene ontology enrichment analysis for genes with most variable expression. (A) Top 3000 genes unique to scRNAseq (i.e. 1520 genes). (B) Top 3000 genes unique to snRNAseq (i.e. 1520 genes). (C) Top 3000 genes shared between scRNAseq and snRNAseq (i.e. 1480 genes). See also Supplementary Table S9 for a full list of enriched GO terms. Fig. S9. Comparison of dataset specific and shared differentially expressed genes for each cell type. The radar plot shows for each cell type the percentage of cluster specific differentially expressed genes unique for the scRNAseq and snRNAseq data, respectively (red and blue lines), as well as the percentage of differentially expressed genes shared between both datasets (black line). The total number of genes fulfilling the differential expression criteria (FDR 0.05 and log2-fold change > 0.25) for each cell type is shown in brackets. Fig. S10. Clustering and cluster annotation of integrated scRNAseq and snRNAseq dataset. (A) The heatmap shows the score for each potential cell type (Y-axis) in each cluster (X-axis). The cell types are annotated based on the highest scoring identity in the heatmap. The clusters are grouped based on their transcriptional similarity to each other. For clusters which express an equal number of marker genes for two different identities both identities were assigned (e.g. cluster 14:Dorsal|Ocelli). Clusters with unresolved identities (i.e. more than two equal assignments) are called “Other”. The colors of the cluster names correspond to the colors in the UMAP in (B). The marker score is calculated using a matrix of published marker genes (see Additional file Table S2). (B) UMAP of integrated scRNAseq and snRNAseq data. Cells are colored by clusters identified based on the (A). Note that the color code in A and B is not comparable to the one used in Additional file Figs. S4 and S7. Table S1. Overview of different dissociation conditions. Samples within blocks (highlighted in grey and white) were prepared in parallel. The Flow Cytometer only provides percentages of survival because it stops after a defined number of events (i.e. ~50,000 cells) and therefore absolute numbers are not meaningful. “Pipetting” refers to the number of strokes during and after incubation. The cells obtained by experiment/block 12 were subjected to 10X Genomics scRNAseq.

Additional file 1: Figure S1. A kinome-wide CRISPR screen in pancreatic cancer cells. Figure S2. Targeted inhibition of CDK7 enhances gemcitabine and paclitaxel chemotherapy response in pancreatic cancer in vitro and in vivo. Figure S3. CDK7 inhibition induces cell cycle arrest, apoptotic cell death, and DNA damage. Figure S4. Targeted inhibition of CDK7 reverses chemoresistance in pancreatic cancer. Table S1. Cell lines. Table S2. Plasmids and primers list. Table S3. Antibody list. Table S4. List of CRISPR-Cas9 screening results of gemcitabine and paclitaxel in TB32047 cells.

BACKGROUND AND AIMS: Non-alcoholic fatty liver disease (NAFLD) is initiated by steatosis and can progress via fibrosis and cirrhosis to hepatocellular carcinoma (HCC). The RNA binding protein HuR controls RNAs at the posttranscriptional level; hepatocyte HuR has been implicated in the regulation of diet-induced hepatic steatosis. The present study aimed to understand the role of hepatocyte-HuR in NAFLD development and progression to fibrosis and HCC. APPROACH AND RESULTS: Hepatocyte-specific HuR-deficient mice and control HuR-sufficient mice were fed either a normal diet or a NAFLD-inducing diet. Hepatic lipid accumulation, inflammation, fibrosis and HCC development were studied by histology, flow cytometry, quantitative PCR and RNA sequencing. The liver lipidome was characterized by lipidomics analysis and the HuR-RNA interactions in the liver were mapped by RNA immunoprecipitation-sequencing. Hepatocyte-specific HuR-deficient mice displayed spontaneous hepatic steatosis and fibrosis predisposition, compared to control HuR-sufficient mice. On a NAFLD-inducing diet, hepatocyte-specific HuR-deficiency resulted in exacerbated inflammation, fibrosis and HCC-like tumor development. A multi-omic approach, including lipidomics, transcriptomics and RNA-immunoprecipitation sequencing revealed that HuR orchestrates a protective network of hepatic-metabolic and lipid homeostasis-maintaining pathways. Consistently, HuR-deficient livers accumulated, already at steady-state, a triglyceride signature resembling that of NAFLD livers. Moreover, upregulation of Spp1 and its product osteopontin mediated, at least partially, the fibrosis development in hepatocyte-specific HuR deficiency on a NAFLD-inducing diet, as shown by experiments utilizing antibody blockade of osteopontin. CONCLUSIONS: HuR is a gatekeeper of liver homeostasis preventing NAFLD-related fibrosis and HCC, suggesting that the HuR-dependent network could be exploited therapeutically.

Background. Adolescence hosts a sharp increase in the incidence of mental disorders. The prodromal phases are often characterized by cognitive deficits which predate disease onset by several years. Characterization of cognitive performance in relation to normative trajectories may have value for early risk assessment and monitoring. Methods. Youth aged 8-21 years (n=6481) from the Philadelphia Neurodevelopmental Cohort (PNC) were included. Performance scores from a computerized neurocognitive battery were decomposed using principal component analysis, yielding a general cognitive score. Items reflecting various aspects of psychopathology from self-report questionnaires and collateral caregiver information were decomposed using independent component analysis, providing individual domain scores. Using normative modeling and Bayesian statistics we estimated normative trajectories of cognitive function and tested for associations between cognitive deviance and psychopathological domain scores. In addition, we tested for associations with polygenic scores for mental and behavioral disorders often involving cognition, including schizophrenia, bipolar disorder, attention deficit hyperactivity disorder (ADHD) and Alzheimer’s disease. Results. More negative normative cognitive deviations were associated with higher general psychopathology burden and domains reflecting positive and prodromal psychosis, attention problems, norm-violating behavior, and anxiety. In addition, better performance was associated with higher joint burden of depression, suicidal ideation, and negative psychosis symptoms. The analyses revealed no evidence for associations with polygenic scores. Conclusion. Our results show that cognitive performance is associated with general and specific domains of psychopathology in youth. These findings support the close links between cognition and psychopathology in youth and highlight the potential of normative modeling for early risk assessment.

The odontoblast, with processes beyond the dentin-enamel boundary, is the initial site of TRPC5 cold transduction in teeth., Teeth are composed of many tissues, covered by an inflexible and obdurate enamel. Unlike most other tissues, teeth become extremely cold sensitive when inflamed. The mechanisms of this cold sensation are not understood. Here, we clarify the molecular and cellular components of the dental cold sensing system and show that sensory transduction of cold stimuli in teeth requires odontoblasts. TRPC5 is a cold sensor in healthy teeth and, with TRPA1, is sufficient for cold sensing. The odontoblast appears as the direct site of TRPC5 cold transduction and provides a mechanism for prolonged cold sensing via TRPC5’s relative sensitivity to intracellular calcium and lack of desensitization. Our data provide concrete functional evidence that equipping odontoblasts with the cold-sensor TRPC5 expands traditional odontoblast functions and renders it a previously unknown integral cellular component of the dental cold sensing system.

Background: Human beings have a basic need for healthy social connections. In our everyday lives, being rejected or excluded when we try to establish or maintain social connections produces negative affect. However, when studied in the laboratory, psychologists rarely find that social exclusion or social rejection induces a negative affective response. Objectives: To better explain the discrepancy between real-life and laboratory findings, three key issues with tasks used to induce feelings of exclusion or rejection are identified: believability, participant engagement and the meaningfulness of the exclusion or rejection. Here, I present a novel social feedback paradigm designed to be believable, engage participants and induce meaningful rejection-related negative affect. Methods: The task was designed to resemble an online dat- ing app and implemented in Psychopy software. Participants engaged with the task in three separate parts: 1) they created their own profile consisting of a selfie and an introductory text. 2) They viewed and rated similar ‘profiles’ of men and/or women (depending on their prefer- ence) that I had created. 3) In the social feedback part of the task, participants received purported ‘feedback’ from others, both rejection and acceptance. Task validation: I created and ran a poll to determine suitable adjectives for the rejection and acceptance feedback. Fourteen participants who participated in a larger mixed-design fMRI psychopharmacological pilot study completed the task. A separate larger behavioral pilot of the task was planned, but cancelled due to COVID 19-restrictions. Results: Analyses revealed that participant felt worse receiving negative feedback compared to positive feedback, but the average affective re- sponse to negative feedback was still close to zero on a -10 to 10 scale. Descriptive analyses did not identify any strong moderators. Conclusion: A small pilot study of a novel task failed to produce an affective response that was substantially stronger than existing tasks. Further investigation is needed to properly assess if the differences between real world and laboratory findings are explainable by believability, participant engagement or meaningfulness.

Pancreatic cancer has become the third leading cause of cancer-related death in the Western world despite advances in therapy of other cancerous lesions. Late diagnosis due to a lack of symptoms during early disease allows metastatic spread of the tumor. Most patients are considered incurable because of metastasized disease. On a cellular level, pancreatic cancer proves to be rather resistant to chemotherapy. Hence, early detection and new therapeutic targets might improve outcomes. The detection of DNA promoter hypermethylation has been described as a method to identify putative genes of interest in cancer entities. These genes might serve as either biomarkers or might lead to a better understanding of the molecular mechanisms involved. We checked tumor specimens from 80 patients who had undergone pancreatic resection for promoter hypermethylation of the zinc finger protein ZNF154. Then, we further characterized the effects of ZNF154 on cell viability and gene expression by in vitro experiments. We found a significant association between ZNF154 hypermethylation and better survival in patients with resectable pancreatic cancer. Moreover, we suspect that the cell growth suppressor SLFN5 might be linked to a silenced ZNF154 in pancreatic cancer.

Background Reptiles are a species-rich group with great phenotypic and life history diversity but are highly underrepresented among the vertebrate species with sequenced genomes. Results Here, we report a high-quality genome assembly of the tegu lizard, Salvator merianae, the first lacertoid with a sequenced genome. We combined 74X Illumina short-read, 29.8X Pacific Biosciences long-read, and optical mapping data to generate a high-quality assembly with a scaffold N50 value of 55.4 Mb. The contig N50 value of this assembly is 521 Kb, making it the most contiguous reptile assembly so far. We show that the tegu assembly has the highest completeness of coding genes and conserved non-exonic elements (CNEs) compared to other reptiles. Furthermore, the tegu assembly has the highest number of evolutionarily conserved CNE pairs, corroborating a high assembly contiguity in intergenic regions. As in other reptiles, long interspersed nuclear elements comprise the most abundant transposon class. We used transcriptomic data, homology- and de novo gene predictions to annotate 22,413 coding genes, of which 16,995 (76%) likely have human orthologs as inferred by CESAR-derived gene mappings. Finally, we generated a multiple genome alignment comprising 10 squamates and 7 other amniote species and identified conserved regions that are under evolutionary constraint. CNEs cover 38 Mb (1.8%) of the tegu genome, with 3.3 Mb in these elements being squamate specific. In contrast to placental mammal-specific CNEs, very few of these squamate-specific CNEs (

Background Reptiles are a species-rich group with great phenotypic and life history diversity but are highly underrepresented among the vertebrate species with sequenced genomes. Results Here, we report a high-quality genome assembly of the tegu lizard, Salvator merianae, the first lacertoid with a sequenced genome. We combined 74X Illumina short-read, 29.8X Pacific Biosciences long-read, and optical mapping data to generate a high-quality assembly with a scaffold N50 value of 55.4 Mb. The contig N50 value of this assembly is 521 Kb, making it the most contiguous reptile assembly so far. We show that the tegu assembly has the highest completeness of coding genes and conserved non-exonic elements (CNEs) compared to other reptiles. Furthermore, the tegu assembly has the highest number of evolutionarily conserved CNE pairs, corroborating a high assembly contiguity in intergenic regions. As in other reptiles, long interspersed nuclear elements comprise the most abundant transposon class. We used transcriptomic data, homology- and de novo gene predictions to annotate 22,413 coding genes, of which 16,995 (76%) likely have human orthologs as inferred by CESAR-derived gene mappings. Finally, we generated a multiple genome alignment comprising 10 squamates and 7 other amniote species and identified conserved regions that are under evolutionary constraint. CNEs cover 38 Mb (1.8%) of the tegu genome, with 3.3 Mb in these elements being squamate specific. In contrast to placental mammal-specific CNEs, very few of these squamate-specific CNEs (

Intrinsic and acquired resistances are major obstacles in cancer therapy. Genetic characterization is commonly used to identify predictive or prognostic biomarker signatures and potential cancer targets in samples from therapy-naïve patients. By far less common are such investigations to identify specific, predictive and/or prognostic gene signatures in patients or cancer cells refractory to a specific molecular-targeted intervention. This, however, might have a great value to foster the development of tailored, personalized cancer therapy. Based on our identification of a differential radiosensitization by single and combined β1 integrin (AIIB2) and EGFR (Cetuximab) targeting in more physiological, three-dimensional head and neck squamous cell carcinoma (HNSCC) cell cultures, we performed comparative whole exome sequencing, phosphoproteome analyses and RNAi knockdown screens in responder and non-responder cell lines. We found a higher rate of gene mutations with putative protein-changing characteristics in non-responders and different mutational profiles of responders and non-responders. These profiles allow stratification of HNSCC patients and identification of potential targets to address treatment resistance. Consecutively, pharmacological inhibition of mTOR and KEAP1 effectively diminished non-responder insusceptibility to β1 integrin and EGFR targeting for radiosensitization. Our data pinpoint the added value of genetic biomarker identification after selection for cancer subgroup responsiveness to targeted therapies.

Figure S1. Sequencing and assembly strategy of the A. salmonicida subsp. pectinolytica strain 34mel genome. Figure S2. Gene similarity and Mummer alignments to other Aeromonas genomes. Figure S3. Transposon conglomerates from A. salmonicida subsp. pectinolytica strain 34mel. Figure S4. Transposon family content in analyzed Aeromonas strains. Figure S5. Comparison of the A. salmonicida subsp. pectinolytica strain 34mel genome, plasmid pFBAOT6 from Aeromonas caviae, and transposon Tn1721. Table S1. Pathogenic as well as temperature-dependent growth features of analyzed Aeromonas strains. Table S2. General genome features of A. salmonicida subsp. pectinolytica strain 34mel. Table S3. Polymorphic sites in rRNAs from A. salmonicida subsp. pectinolytica strain 34mel. Table S4. Genes disrupted by transposons in A. salmonicida subsp. pectinolytica strain 34mel. Table S5. Transposon details from analyzed Aeromonas strains. Table S6. Genes retained in transposon TnAs1 from the environmental IncP-9 TOL plasmid pWW0 from Pseudomonas putida. Text S1. Annotation details for RNA genes in the A. salmonicida subsp. pectinolytica strain 34mel genome. Text S2. Short unassigned contigs from the A. salmonicida subsp. pectinolytica strain 34mel draft genome. Text S3. Transposons: nomenclature and completeness. Text S4. On transposon counting and the meaning of â ISAs11â . Text S5. Insertions on pFBAOT6 in transposon Tn1721 and details of the regions shared with the A. salmonicida subsp. pectinolytica strain 34mel genome. Text S6. Details of PacBio library preparation, sequencing, and assembly of the genome from A. salmonicida subsp. pectinolytica strain 34mel. (PDF 1171 kb)

Plasmacytoid dendritic cells (pDCs) regulate innate and adaptive immunity. Neurotrophins and their receptors control the function of neuronal tissue. In addition, they have been demonstrated to be part of the immune response but little is known about the effector immune cells involved. We report, for the first time, the expression and immune-regulatory function of the low affinity neurotrophin receptor p75 neurotrophin receptor (p75NTR) by the antigen-presenting pDCs, mediated by toll-like receptor (TLR) 9 activation and differential phosphorylation of interferon regulatory factor 3 and 7. The modulation of p75NTR on pDCs significantly influences disease progression of asthma in an ovalbumin-induced mouse model mediated by the TLR9 signaling pathway. p75NTR activation of pDCs from patients with asthma increased allergen-specific T cell proliferation and cytokine secretion in nerve growth factor concentration-dependent manner. Further, p75NTR activation of pDCs delayed the onset of autoimmune diabetes in RIP-CD80GP mice and aggravated graft-versus-host disease in a xenotransplantation model. Thus, p75NTR signaling on pDCs constitutes a new and critical mechanism connecting neurotrophin signaling and immune response regulation with great therapeutic potential for a variety of immune disorders.

Background Clonal competition in cancer describes the process in which the progeny of a cell clone supersedes or succumbs to other competing clones due to differences in their functional characteristics, mostly based on subsequently acquired mutations. Even though the patterns of those mutations are well explored in many tumors, the dynamical process of clonal selection is underexposed. Methods We studied the dynamics of clonal competition in a BcrAbl-induced leukemia using a γ-retroviral vector library encoding the oncogene in conjunction with genetic barcodes. To this end, we studied the growth dynamics of transduced cells on the clonal level both in vitro and in vivo in transplanted mice. Results While we detected moderate changes in clonal abundancies in vitro, we observed monoclonal leukemias in 6/30 mice after transplantation, which intriguingly were caused by only two different BcrAbl clones. To analyze the success of these clones, we applied a mathematical model of hematopoietic tissue maintenance, which indicated that a differential engraftment capacity of these two dominant clones provides a possible explanation of our observations. These findings were further supported by additional transplantation experiments and increased BcrAbl transcript levels in both clones. Conclusion Our findings show that clonal competition is not an absolute process based on mutations, but highly dependent on selection mechanisms in a given environmental context. Electronic supplementary material The online version of this article (doi:10.1186/s12943-017-0668-x) contains supplementary material, which is available to authorized users.

eGFP expression and clonal kinetics in the BcrAbl_bulk culture. Figure S2: Overlap of barcodes from plasmid library and first sample. Figure S3: Clonal kinetics in the eGFP-positive cultures after sorting. Figure S4: Spleens from diseased (and control) animals. Figure S5: Read counts for Klf6 and Sprouty1. Figure S6: Comparison of samples. Figure S7: KEGG enrichment analysis. Table S1: Spleen weights and eGFP expression in the diseased animals. Supplementary Model Description. Table S2: Model Parameters. Table S3: Oligonucleotides. (PDF 7697Â kb)

Denna studie syftar till att skapa förståelse för hur IKT och digitala verktyg används på fritidshemmet, vidare riktar sig studiens undersökning mot fritidslärarna för att ta del av deras uppfattningar om användning av IKT och digitala verktyg. För att svara på syftet utgår studien från följande frågeställningar: Vad finns det för bakomliggande faktorer i fritidslärarnas användning och vad ser de själva att det finns för för- och nackdelar med användning av IKT och digitala verktyg, hur och varför används digitala verktyg enligt lärarna och hur arbetar lärarna med källkritik och etiska frågor?Fritidshemmet ska enligt styrdokument ge eleverna kunskaper om och i de digitala verktygen, få möjligheter att utforska verktygen samt använda sig av dessa ämnesöverskridande. Vidare ska fritidshemmet fungera som en utjämnande faktor för olika sociala förhållanden. De elever som inte har tillgång till digitala verktyg i hemmet ska alltså erbjudas möjligheter att via fritidshemmet få kunskap som jämnar ut de digitala klyftor som finns. Studien kommer genomföras på två skolor med tre fritidshem. Vidare kommer endast fritidslärare som innehar pedagogisk utbildning vara delaktiga i studien. Datainsamlingen för denna studie har skett genom sex stycken kvalitativa intervjuer. Under intervjuerna har respondenterna fått presentera deras egna åsikter och attityder gentemot IKT och digitala verktyg. Dessa utsagor har därefter ställts mot aktuell forskning som delvis visat liknande resultat som studiens respondenter. Resultatet av det analyserade empiriska materialet påvisar en brist i många av de områden som påverkar arbetet med IKT och digitala verktyg. Främst framkommer det att respondenternas attityder, eget intresse, tillgång av verktygen och stöd från ledningen är de största faktorerna som påverkar denna användning både positivt och negativt. Det som inte påverkade var elevernas egna intressen, viljor eller aktuella styrdokument.

Plasmacytoid dendritic cells (pDCs) regulate innate and adaptive immunity. Neurotrophins and their receptors control the function of neuronal tissue. In addition, they have been demonstrated to be part of the immune response but little is known about the effector immune cells involved. We report, for the first time, the expression and immune-regulatory function of the low affinity neurotrophin receptor p75 neurotrophin receptor (p75NTR) by the antigen-presenting pDCs, mediated by toll-like receptor (TLR) 9 activation and differential phosphorylation of interferon regulatory factor 3 and 7. The modulation of p75NTR on pDCs significantly influences disease progression of asthma in an ovalbumin-induced mouse model mediated by the TLR9 signaling pathway. p75NTR activation of pDCs from patients with asthma increased allergen-specific T cell proliferation and cytokine secretion in nerve growth factor concentration-dependent manner. Further, p75NTR activation of pDCs delayed the onset of autoimmune diabetes in RIP-CD80GP mice and aggravated graft-versus-host disease in a xenotransplantation model. Thus, p75NTR signaling on pDCs constitutes a new and critical mechanism connecting neurotrophin signaling and immune response regulation with great therapeutic potential for a variety of immune disorders.

Utifrån den framtagna frågeställningen ämnar denna rapport att redogöra hur webbapplikationen Vardagsboxen konstruerats för att uppfylla dess vision om användbarhet. Teori, uppkomna resultat och användartester har lett fram till att en användbar webbapplikation grundar sig i hur användaren upplever denna genom bland annat responsivitet och aktiv återkoppling från webbapplikationen. Därtill är färgval samt antalet använda färger viktigt för att främja användbarhet och i förlängningen vara visuellt tilltalande för den tänkta användarskaran. En användbar webbapplikation ska även vara enkel i design och igenkännbar. Arbetssättet Scrum har applicerats för att strukturera upp arbetsgången under olika iterationer där teknisk utveckling varit i fokus. Vardagsboxen är en internetbutik i form av en prenumerationstjänst där konsumenterna kan prenumerera på vardagsartiklar. Rapporten behandlar webbapplikationens uppbyggnad samt skapandet av en lättnavigerad och användbar tjänst med syfte att frigöra tid i vardagen åt dess användare. Med hjälp av en grundlig prototypframtagning i kombination med analyser av bakomliggande tekniska lösningar, design och resultat från användartester har utvecklingsteamet lyckats skapa en användbar webbapplikation. By focusing on the established thesis, the report aims to depict how the web application Vardagsboxen has been constructed concentrating on the aspect of usability. Theory, results and user testing have led to the conclusions that a usable web application is based on how the user experiences this through responsiveness and active feedback. Furthermore, the coloring and the numbers of colors used are important to enhance usability. A usable web application should also be simple in design and recognizable. The methodology of Scrum has been implemented to structure the project during the ongoing iterations. During these iterations, technical development has been the main focus. Vardagsboxen is an internet shop where the consumers can subscribe to packages of hygiene products. The report includes the process of creating an accessible and serviceable internet shop with hopes to assist the consumers in their everyday life. Through a thorough prototype development in combination with analyzing technical solutions, design and results from usability tests, the development team has successfully created a web application while focusing on usability.

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)

LB

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.

April 1, 2011, The 1000 Genomes Project aims to provide a deep characterization of human genome sequence variation as a foundation for investigating the relationship between genotype and phenotype. Here we present results of the pilot phase of the project, designed to develop and compare different strategies for genome-wide sequencing with high-throughput platforms. We undertook three projects: low-coverage whole-genome sequencing of 179 individuals from four populations; high-coverage sequencing of two mother–father–child trios; and exon-targeted sequencing of 697 individuals from seven populations. We describe the location, allele frequency and local haplotype structure of approximately 15 million single nucleotide polymorphisms, 1 million short insertions and deletions, and 20,000 structural variants, most of which were previously undescribed. We show that, because we have catalogued the vast majority of common variation, over 95% of the currently accessible variants found in any individual are present in this data set. On average, each person is found to carry approximately 250 to 300 loss-of-function variants in annotated genes and 50 to 100 variants previously implicated in inherited disorders. We demonstrate how these results can be used to inform association and functional studies. From the two trios, we directly estimate the rate of de novo germline base substitution mutations to be approximately 10[superscript −8] per base pair per generation. We explore the data with regard to signatures of natural selection, and identify a marked reduction of genetic variation in the neighbourhood of genes, due to selection at linked sites. These methods and public data will support the next phase of human genetic research., Ministry of Science and Technology of the People's Republic of China (973 program no. 2011CB809200), National Natural Science Foundation (China) (30725008), National Natural Science Foundation (China) (30890032), National Natural Science Foundation (China) (30811130531), National Natural Science Foundation (China) (30221004), China (Chinese 863 program 2006AA02Z177), China (Chinese 863 program 2006AA02Z334), China (Chinese 863 program 2006AA02A302), China (Chinese 863 program 2009AA022707), National Institutes of Health (U.S.) (Grant U54HG2750), National Institutes of Health (U.S.) (Grant U01HG5208), National Institutes of Health (U.S.) (Grant U54HG3067)

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.

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. 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 experiments using sanding material, the unwashed natural sand caused the highest PM10 concentrations. Friction tyres on natural sand caused higher concentrations than studded tyres on washed crushed stone. Studded tyres generated more PM10 than friction tyres but the difference was not as large as when no sanding material was used. 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.

Conditional gene inactivation is a powerful tool to determine gene function when constitutive mutations result in detrimental effects. The most commonly used technique to achieve conditional gene inactivation employs the Cre/loxP system and its ability to delete DNA sequences flanked by two loxP sites. However, targeting a gene with two loxP sites is time and labor consuming. Here, we show Cre-Controlled CRISPR (3C) mutagenesis to circumvent these issues. 3C relies on gRNA and Cre-dependent Cas9-GFP expression from the same transgene. Exogenous or transgenic supply of Cre results in Cas9-GFP expression and subsequent mutagenesis of the gene of interest. The recombined cells become fluorescently visible enabling their isolation and subjection to various omics techniques. Hence, 3C mutagenesis provides a valuable alternative to the production of loxP-flanked alleles. It might even enable the conditional inactivation of multiple genes simultaneously and should be applicable to other model organisms amenable to single integration transgenesis., Targeting a gene with two loxP sites is both time and labour intensive. Here the authors present Cre-Controlled CRISPR allowing conditional mutagenesis of a gene of interest and simultaneously labelling the putative mutant cells fluorescently.

Trained innate immunity fosters a sustained favorable response of myeloid cells to a secondary challenge, despite their short lifespan in circulation. We thus hypothesized that trained immunity acts via modulation of hematopoietic stem and progenitor cells (HSPCs). Administration of β-glucan (prototypical trained-immunity-inducing agonist) to mice induced expansion of progenitors of the myeloid lineage, which was associated with elevated signaling by innate immune mediators, such as IL-1β and granulocyte-macrophage colony-stimulating factor (GM-CSF), and with adaptations in glucose metabolism and cholesterol biosynthesis. The trained-immunity-related increase in myelopoiesis resulted in a beneficial response to secondary LPS challenge and protection from chemotherapy-induced myelosuppression in mice. Therefore, modulation of myeloid progenitors in the bone marrow is an integral component of trained immunity, which to date, was considered to involve functional changes of mature myeloid cells in the periphery., Cell, 172 (1-2), ISSN:0092-8674, ISSN:1097-4172

[Background]: Morphology of the human cerebral cortex differs across psychiatric disorders, with neurobiology and developmental origins mostly undetermined. Deviations in the tangential growth of the cerebral cortex during pre/perinatal periods may be reflected in individual variations in cortical surface area later in life., [Methods]: Interregional profiles of group differences in surface area between cases and controls were generated using T1-weighted magnetic resonance imaging from 27,359 individuals including those with attention-deficit/hyperactivity disorder, autism spectrum disorder, bipolar disorder, major depressive disorder, schizophrenia, and high general psychopathology (through the Child Behavior Checklist). Similarity of interregional profiles of group differences in surface area and prenatal cell-specific gene expression was assessed., [Results]: Across the 11 cortical regions, group differences in cortical area for attention-deficit/hyperactivity disorder, schizophrenia, and Child Behavior Checklist were dominant in multimodal association cortices. The same interregional profiles were also associated with interregional profiles of (prenatal) gene expression specific to proliferative cells, namely radial glia and intermediate progenitor cells (greater expression, larger difference), as well as differentiated cells, namely excitatory neurons and endothelial and mural cells (greater expression, smaller difference). Finally, these cell types were implicated in known pre/perinatal risk factors for psychosis. Genes coexpressed with radial glia were enriched with genes implicated in congenital abnormalities, birth weight, hypoxia, and starvation. Genes coexpressed with endothelial and mural genes were enriched with genes associated with maternal hypertension and preterm birth., [Conclusions]: Our findings support a neurodevelopmental model of vulnerability to mental illness whereby prenatal risk factors acting through cell-specific processes lead to deviations from typical brain development during pregnancy., This work was supported by the German Research Foundation (DFG Grant Nos. HA7070/2-2, HA7070/3, HA7070/4 [to TH]) and IZKF of the medical faculty of Münster (Grants No. Dan3/012/17 [to UD] and MzH 3/020/20 [to TH]), and NHMRC projects (Grant No. 1064643 [to BJH] and 1024570 [to CGD]). The CIAM study was supported by the University of Cape Town Research Committee, South African National Research Foundation, and the South African Medical Research Council (principal investigator [PI], Fleur M. Howells) and Grant No. R01MH117601 (to NJ). This work was funded by the German Research Foundation (Grant Nos. FOR2107 JA 1890/7-1 and FOR2107 JA 1890/7-2 [to AJ]) and Swinburne University scholarship/Australian Postgraduate Award (to JK). Collection of the COGSBD cohort was funded by the Jack Brockhoff Foundation, University of Melbourne, Barbara Dicker Brain Sciences Foundation, Rebecca L Cooper Foundation, and the Society of Mental Health Research (to JK). This work was funded by the German Research Foundation (Grant Nos. FOR2107 KI588/14-1 and FOR2107 KI588/14-2 [to TK]). The St. Göran study was supported by grants from the Swedish Research Council (Grant No. 2018-02653 [to ML]), the Swedish foundation for Strategic Research (Grant No. KF10-0039 [to ML]), the Swedish Brain foundation (Grant No. FO2020-0261 [to ML]), and the Swedish Government under the LUA/ALF agreement (Grant Nos. ALF 20170019 and ALFGBG-716801 [to ML]). This work was supported by RFBR (Grant No. 20-013-00748 [to IL, AST]) and funded by the Health Research Board (Grant No. HRA_POR/2011/100 [to CM]). The Australian Schizophrenia Research Bank (ASRB) was supported by the NHMRC (Enabling Grant No. 386500), the Pratt Foundation, Ramsay Health Care, the Viertel Charitable Foundation, and the Schizophrenia Research Institute. Chief investigators for ASRB were VC, US, RSc, AJ, BM, PTM, SVC, FH, and CPa. This work was supported by Deutsche Forschungsgemeinschaft (Grant Nos. DFG NE 2254/2-1, NE 2254/3-1, NE2254/4-1 [to IN]), the University Research Priority Program “Integrative Human Physiology” at the University of Zurich (to ROT), an NHMRC Senior Principal Research Fellowship (Grant No. 1105825 [to CPa]), an NHMRC L3 Investigator Grant (Grant No. 1196508 [to CPa]), and NHMRC Program Grant (Grant No. 1150083 [to CPa]). ASRB was supported by the NHMRC (Enabling Grant No. 386500), the Pratt Foundation, Ramsay Health Care, the Viertel Charitable Foundation and the Schizophrenia Research Institute. Chief investigators for ASRB were VC, US, RSc, AJ, BM, PTM, SVC, FH, and CPa. This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant (Agreement No. 798181 [to GP]). ASRB was supported by the NHMRC (Enabling Grant, ID 386500), the Pratt Foundation, Ramsay Health Care, the Viertel Charitable Foundation and the Schizophrenia Research Institute. Chief investigators for ASRB were VC, US, RSc, AJ, BM, PTM, SVC, FH, and CPa. The Imaging Genetics in Psychosis study was funded by Project Grants from the NHMRC (Grant Nos. APP630471 and APP1081603 [to YQ]) and the Macquarie University’s Australian Research Council (ARC) Centre of Excellence in Cognition and its Disorders (Grant No. CE110001021 [to YQ]). This work was supported by the Spanish Ministry of Science, Innovation and Universities/Economy and Competitiveness/Instituto de Salud Carlos III (Grant Nos. PI11/01766 and CPII19/00009 [to JR]), co-financed by European Regional Development Fund funds from the European Commission (“A Way of Making Europe”). ASRB was supported by the NHMRC (Enabling Grant No. 386500), the Pratt Foundation, Ramsay Health Care, the Viertel Charitable Foundation, and the Schizophrenia Research Institute. Chief investigators for ASRB were VC, US, RSc, AJ, BM, PTM, SVC, FH, and CPa. SR was supported by an NHMRC Senior Fellowship (Grant No. GNT1154651). Collection of the COGSBD cohort was funded by the Jack Brockhoff Foundation, University of Melbourne, Barbara Dicker Brain Sciences Foundation, Rebecca L Cooper Foundation, and the Society of Mental Health Research (to SR). This work was supported by NIHR; MRC (to KR), NIH (Grant Nos. MH-094268, MH-105660, and MH-107730 [to ASS]). The Neuroimaging of the Children's Attention Project cohort was funded by NHMRC, Australia (Grant No. 1065895). Earlier funding for the cohort as also provided by NHMRC (Grant No. 1008522) and a grant from the Collier Foundation. The ACPU cohort was funded by NHMRC, Australia (Project Grant Nos. 384419 and 569533 [to TS]). This work was supported by research grants from the National Healthcare Group, Singapore (Grant Nos. SIG/05004, SIG/05028, and SIG /1103), and the Singapore Bioimaging Consortium (RP C009/2006 [to KS]), and the Ministry of Health, Czech Republic - conceptual development of research organization (“Institute for Clinical and Experimental Medicine - IKEM, IN 00023001” [to ASk]). This study was supported by the Italian Ministry of Health (Grant No. RC/17-18-19-20-21/A [to GS]) and Ministry of Health of the Czech Republic (Grant No. NU20-04-00393 [to FS]). The Drakenstein Child Health Study (DCHS) cohort is funded by the Bill and Melinda Gates Foundation (Grant No. OPP 1017641) and the South African Medical Research Council. This DCHS contribution was made possible in part by a grant from Carnegie Corporation of New York. The statements made and views expressed are solely the responsibility of the author (DJS). STRADL study was supported and funded by the Wellcome Trust Strategic Award “Stratifying Resilience and Depression Longitudinally” (Grant No. 104036/Z/14/Z), and the Medical Research Council Mental Health Pathfinder Award “Leveraging routinely collected and linked research data to study the causes and consequences of common mental disorders” (MRC, Grant No. MC_PC_17209). Scottish Bipolar Family Study (SBFS) was supported by National Health Service Research Scotland, through the Scottish Mental Health Research Network (www.smhrn.org.uk), who provided assistance with subject recruitment and assessments. SBFS was conducted at the Brain Research Imaging Centre (http://www.bric.ed.ac.uk), which is supported by SINAPSE (Scottish Imaging Network, a Platform for Scientific Excellence, http://www.sinapse.ac.uk). Processing of the datasets used the resources provided by the Edinburgh Compute and Data Facility (http://www.ecdf.ed.ac.uk/) (ASt). TVR was supported by an NHMRC Early Career Fellowship (Grant No. GNT1088785). Collection of the COGSBD cohort was funded by the Jack Brockhoff Foundation, University of Melbourne, Barbara Dicker Brain Sciences Foundation, Rebecca L Cooper Foundation, and the Society of Mental Health Research (to TVR). EV was supported by the Spanish Ministry of Science and Innovation (PI18/00805) integrated into the Plan Nacional de I+D+I and co-financed by the ISCIII-Subdirección General de Evaluación and the FEDER; the Instituto de Salud Carlos III; the CIBERSAM (Centro de Investigación Biomédica en Red de Salud Mental); by the CERCA Programme/Generalitat de Catalunya and the Secretaria d’Universitats i Recerca del Departament d’Economia I Coneixement (Grant No. 2017SGR1355). This study was also supported by the Departament de Salut de la Generalitat de Catalunya, Pla Estratègic de Recerca i Innovació en Salut (PERIS) 2016-2020 (Grant No. SLT006/17/00345) and the European Union Horizon 2020 research and innovation program (EU.3.1.1. Understanding health, wellbeing and disease: Grant Nos. 754907 and EU.3.1.3 [to EB]; Treating and managing disease: Grant No. 945151 [to EV]). This study was supported by the National Center for Complementary and Integrative Health (Grant Nos. R21AT009173 and R61AT009864 [to TTY]); by the National Center for Advancing Translational Sciences (CTSI), National Institutes of Health, through UCSF-CTSI (Grant No. UL1TR001872 [to TTY]); by the American Foundation for Suicide Prevention (Grant No. SRG-1-141-18 [to TTY]); by UCSF Research Evaluation and Allocation Committee (REAC) and J. Jacobson Fund (to TTY); by the NIMH (Grant No. R01MH085734 [to TTY]); and by the Brain and Behavior Research Foundation (formerly NARSAD) (to TTY). This work was supported by a personal Veni grant to MH from the Netherlands Organization for Scientific Research (NWO, Grant No. 91619115 [to MH]) and European Community’s Horizon 2020 Programme (H2020/2014 – 2020) (Grant Agreements Nos. 667302 [CoCA], 728018 [Eat2beNICE], and 847879 [PRIME] [to BF]). JBu has been supported by the EU-AIMS (European Autism Interventions) and AIMS-2-TRIALS programmes, which receive support from Innovative Medicines Initiative Joint Undertaking Grant Nos. 115300 and 777394, the resources of which are composed of financial contributions from the European Union’s FP7 and Horizon 2020 Programmes, and from the European Federation of Pharmaceutical Industries and Associations companies’ in-kind contributions, and AUTISM SPEAKS, Autistica and SFARI; and by the Horizon 2020–supported programme CANDY (Grant No. 847818 [to JBu]). This work is supported by Grant No. NIA T32AG058507 and NIH Grant No. U54EB020403 from the Big Data to Knowledge (BD2K) Program (to CRKC). ENIGMA MDD work is supported by NIH (Grant Nos. U54 EB020403 [to PT], R01 MH116147 [to PT], and R01 MH117601 [to NJ and LS]). LS was supported by an NHMRC Career Development Fellowship (Grant No. 1140764). This work was supported by the National Center for Research Resources at the NIH (Grant Nos. NIH 1 U24 RR021992 [Function Biomedical Informatics Research Network], NIH 1 U24 RR025736-01 [Biomedical Informatics Research Network Coordinating Center; http://www.birncommunity.org]). TGMvE is supported by ENIGMA’s NIH BD2K initiative (Grant No. U54 EB020403), ENIGMA Sex Differences (Grant No. R01MH116147), and ENIGMA-COINSTAC: Advanced Worldwide Transdiagnostic Analysis of Valence System Brain Circuits (Grant No. R01MH121246). This work was supported by the NIH (Grant No. R01MH121246 [to JT, Calhoun, and TGMvE]). This work was supported in part by NIH (Grant No. U54 EB020403 [to PT]).

Den maritime næringen står for omtrent 3% av de globale CO2-utslippene. Med et økende fokus på mer miljøvennlige løsninger og et ønske om å løse dagens klimautfordringer, er hybridelektriske skip essensielle for at den maritime næringen skal klare å redusere utslippene sine. Det å installere batterier på skip er derfor et viktig tiltak for at skipsfarten skal bevege seg i en grønnere retning. Denne avhandlingen presenterer en ny modell for hybridelektriske kraftsystemer, som kalles den strukturbevarende modellen (SPM). SPM-en gjelder opprinnelig kun for konvensjonelle kraftsystemer uten batteri, men et av bidragene i denne avhandlingen er å utvide SPM-en for hybridelektriske kraftsystemer, der en batteri-modell er utviklet. Gjennom en verifikasjonsstudie bekreftes det at SPM-en er en nøyaktig modell for hybridelektriske kraftsystemer, og det konkluderes med at SPM-en er godt egnet til kontrollformål, og at den kan brukes som en modell for å optimalisere energibruken i et hybridelektrisk kraftsystem. Optimaliseringsproblemer som bruker modellprediktiv regulering (MPC) er formulert for tre forskjellige batteristrategier, og løsningene på disse problemene presenteres i en casestudie for et dynamisk posisjonert fartøy. De ulike batteristrategiene som brukes i denne avhandlingen er topputjevning (peak-shaving), kraftutjevning (power smoothing) og strategisk lasting (strategic loading), og i tillegg brukes en strategi uten batteri til sammenligning, som representerer et tradisjonelt kraftsystem. Resultatene fra casestudien indikerer at strategisk lasting er en effektiv batteristrategi. Sammenlignet med å ikke bruke batteri, reduserer strategisk lasting drivstofforbruket med 7% for en kort simulering på 1000 sekunder, og med 5,3% for en lang simulering på 24 timer, som er basert på en realistisk fartøysoperasjon. Selv om det ikke er hovedfokuset i denne avhandlingen, sammenlignes også resultatene med en annen effektiv strategi for å redusere utslipp, såkalt gensettfrakobling, som betyr å slå av og på generatorsett avhengig av lasten. Gensettfrakobling gir store drivstoffbesparelser, og resultatene indikerer at for den korte simuleringen reduseres drivstofforbruket med 16,1% for gensettfrakobling uten batteri, og med 17,7% for gensettfrakobling kombinert med strategisk lasting. For den lange simuleringen er de tilsvarende tallene henholdsvis 38,1% og 38,5%. Følgelig anses det å kombinere gensettfrakobling og strategisk lasting som en effektiv strategi for hybridelektriske skip. Simuleringene viser også at transiente effekter kan oppstå i det hybridelektriske kraftsystemet, enten på grunn av batteriet, som antas å levere effekt øyeblikkelig, eller på grunn av ustabilitet knyttet til gensettfrakobling. Når gensettfrakobling ikke utføres, er frekvensen innenfor de stasjonære grensene satt av klasseselskap, men simuleringene viser at gensettfrakobling kan føre til at disse frekvensgrensene overskrides. The maritime industry accounts for approximately 3% of the global CO2 emissions. With the rising concern for more environmentally friendly solutions and a desire to solve today's climate challenges, hybrid electric ships are essential for the maritime industry to reduce its emissions, and installation of batteries on ships is therefore an important measure for shipping to move in a greener direction. This thesis presents a new model for hybrid electric power systems, which is called the structure-preserving model (SPM). The SPM is originally only valid for conventional power systems without battery, but one of the contributions of this thesis is to extend the SPM for hybrid electric power systems, where a battery model is developed. It is verified that the SPM is an accurate model for hybrid electric power systems through a verification study, and it is concluded that the SPM is well fit for control purposes, and that it can be used as a model to optimize the energy use in a hybrid electric power system. Optimization problems using model predictive control (MPC) are formulated for three different battery strategies, and the solutions to these problems are presented in a case study for a dynamically positioned vessel. The different battery strategies in this thesis are using the battery for peak-shaving, power smoothing and strategic loading, and in addition, a strategy without battery is used for comparison, representing a traditional power system. The results from the case study indicate that strategic loading is an efficient battery strategy. Compared to without using battery, strategic loading reduces the fuel consumption with 7% for a short simulation of 1000 seconds, and with 5.3% for a long simulation of 24 hours, based on a realistic vessel operation. Even though not the main focus of this thesis, the results are also compared to another efficient strategy for reducing emissions, which is genset disconnection, meaning turning on and off generator sets depending on the load. Genset disconnection gives large fuel savings, and the results indicate that for the short simulation, the fuel consumption is reduced by 16.1% for genset disconnection without battery, and by 17.7% for genset disconnection combined with strategic loading. For the long simulation, the corresponding numbers are 38.1% and 38.5%, respectively. Consequently, combining genset disconnection and strategic loading is deemed an efficient strategy for hybrid electric ships. The simulations also show that transient effects can occur in the hybrid electric power system, either due to the battery power, which is assumed to be delivered instantly, or due to instabilities from the genset disconnection. When not performing genset disconnection, the frequency is within the steady-state limits set by class societies, but the simulations show that genset disconnection can make the frequency exceed these limits.

The purpose of this thesis is to investigate whether an inverted pendulum can be balanced on a hydrodynamic platform actuated by thrusters. The vessel is the omnidirectional CS Saucer actuated by three rotating azimuth thrusters. The vessel have been heavily modified, replacing the software control system with a Robot Operating System (ROS) based platform, using an Arduino Mega embedded circuit board, and the Raspberry Pi 2. This is a versatile platform, especially useful with MATLAB and Simulink simulations, and for implementation of additional hardware. For this project, rpm measurements from all motors have been added, along with battery voltage monitoring and encoder reading. The encoder is used to read the position of the pendulum. A completely new payload system, interfacing the pendulum and encoder to the CS Saucer have been designed and constructed. The system is designed to only rotate in one plane, and the pendulum is restricted to move in the range $\pm 45\degree$. In order to design a control system that keeps the pendulum at the upright equilibrium, a mathematical model of the system is needed. Equations for an inverted pendulum on a cart, and hydrodynamic modelling of marine vessels are presented, and then combined to find the equations for the system. The Lagrangian approach is used to derive the dynamic coupled equations for the marine inverted pendulum, where theory on the double inverted pendulum is used for comparison. It is also shown that the Denavit-Hartenberg convention used in robotics can be applied on the marine inverted pendulum to derive the equations. The mathematical model is highly coupled and nonlinear. For controller and observer designs, however, it is convenient to work with a simplified model. Therefore, the system is linearized about the desired upright equilibrium and position, i.e. the zero state, and written on state space form. Based on this linear model, a linear quadratic regulator (LQR) feedback controller is designed. Only the vessel position, pitch and pendulum angle are measured, while their derivatives also form part of the state. Since all six states are necessary for feedback control, a Luenberger state estimator is designed. Together with the LQR they form the linear quadratic Gaussian (LQG). The observer is then a combined filter and state estimator, able to reconstruct unmeasured states, and filter noisy measurements. The system is tested in the MC Lab, where real conditions as unreliable sensors, signal losses and noise are present. Experiments show promising results for the LQR. Several runs are conducted, showing that both the control law, and vessel should be suitable for the marine inverted pendulum. The objective is not completely achieved, but it is believed that it is possible to achieve it given some more work. One of the main concerns in this regard is the thruster mapping. The thrusters are unreliable, especially in the zero thrust region, and when thrust direction is changed. Therefore, this should be the main focus area for further work. Moreover, the vessel design might not be the best for a marine inverted pendulum. It is directionally unstable, and a strong pitching effect is observed when thruster force is applied. So, while the omnidirectional property might be very suitable for a freely moving pendulum, a long and thin vessel is believed to be more suitable for the in-plane rotating pendulum.

This thesis presents the development of a new research foundation into the Marine Cybernetic Laboratory, the C/S Inocean Cat I Drillship. This is a 1:90 scaled model of an Arctic drillship design by Inocean for Statoil. The C/S Inocean Cat I Drillship model is equipped with six Aero-naut Precision Schottel azimuth thrusters which are driven by six O.S. OMA-2820-950 motors and six Dynamixel MX-106R servo motors. To control the model a real time controller, CompactRIO, from National Instruments is used. The system is powered by six 12 V 12 Ah lead batteries. To make the hull durable and lightweight, it is constructed by carbon fiber and a casted frame stiffens the hull. The model has a detachable lid made of Plexiglass, that secures the equipment inside from water. In order to design a thruster-assisted position mooring control system, a 6 DOF mathematical model of the scale model is needed. Equations for hydrodynamic modeling of marine vessels and mooring lines are presented, and combined to get the system equations. In addition, a 3 DOF mathematical model has been derived to verify other experiments. The scaled model is tested in the MC Lab, where real scaled conditions can be applied. The experiments show great results for the different controllers, and the vessel manages the different conditions. Comparison between the simulation model and the scale model is performed for all cases. The results yield that the simulation model needs further development to be more similar to the scale model, but it gives a good indication on the behavior of the vessel. The complete C/S Inocean Cat I Drillship is a fully functional vessel, and a stable research foundation to be used in further experiments in the MC Lab.

This thesis reviews the complete design of a control system on a marine surface vessel, capable of autonomous exploration in small-scale marine environments. This involves the development of strategies for map exploration, path planning, navigation, and motion control. Further, the thesis describes how these components are merged into one autonomous system. The experimental platform utilized in this project is the CS Saucer, a model-scale vessel built for testing in the Marine Cybernetics Laboratory at NTNU. The vessel has been extensively upgraded during this project. This includes the installation of new hardware and software, and significant improvements in the vessels capabilities of track following. The system is installed on the Robot Operating System, a flexible platform with a large open-source community. This platform has made it possible to implement tools familiar within the robotics community such as algorithms for performing simultaneous localization and mapping. %The maps that are generated from these algorithms are occupancy-grids, where each cell in the grid is recognized as either a free, occupied or unexplored. Two strategies for exploration are considered in the thesis, where the Frontier Based Exploration strategy is the preferred one. In this strategy the vessel always moves to the edges between known and unknown area. A version of the A* search algorithm has been implemented, responsible for planning paths to locations within the vessels environment. This algorithm is implemented such that node connections may span more than one cell, and with a scheme for weighting cells such that the vessel keeps a distance from walls. The generated paths are in general found to be satisfying. A velocity control law that generates controller setpoints for the motion controller based on planned path and distance to nearby objects has been introduced to the system. The software components responsible for this operation iterates much faster than desired paths are recalculated. In this manner, the system is able to generate a steady stream of setpoints for the motion controller. The resulting system has been tested through simulations, and subsequently verified in experiments performed in a basin facility. The experiments are well documented and are presented in a separate chapter of the thesis. A video demonstrating the successful experiments is referenced in the main body of the thesis. An interface where the operator can interrupt the exploration process and direct the vessel to any position in the explored map has also been created and successfully tested.

This thesis presents the design, construction and control systems development of the marine cybernetics vessel CS Saucer. CS Saucer is a fully actuated and highly controllable vessel with a spherically shaped hull, much like a flying saucer. The vessel is fitted with three azimuth thrusters which are driven by three Torpedo 800 motors. The embedded controller myRIO from National Instruments is used to control the vessel. The system is powered by a three cell 11.1V Lithium Polymer battery. The hull of the vessel is constructed from three millimeter MDF sheeting, milled Divinycell foam and a woven carbon sheet fitted to the exterior of the vessel with epoxy. This results in a rigid and watertight hull with a low draft. The vessel has a detachable lid made of plexi glass that is secured to the top of the vessel with four butterfly screws. The drag forces acting on the vessel have been analyzed through testing in the Marine Cybernetics Laboratory (MC Lab) at NTNU. The results have been used to derive the parameters for a dynamic model used in Hardware-in- the-loop testing and for future model based control design. The forces produced by each of the thrusters have been mapped and approximated with linear curve fitting. This approximated force mapping has been used in the implemented thrust allocation with fixed thruster angles. Manual thruster control and manual force control of the vessel can be operated by the application on the host computer or directly from the Dashboard app from National Instruments available on Android and iOS devices. A dynamic positioning system has been implemented on the CS Saucer and tested in the MC Lab. The dynamic positioning system performs as desired when changing the setpoint for the vessel position. A tentative path following controller has also been implemented and tested on the vessel. The completed CS Saucer is a fully functioning vessel and a stable and expandable platform for use in future demonstrations, projects and research at the MC Lab.