HMGB1 : regulation of inflammatory functions and therapeutic blockade

The intracellular protein High Mobility Group Box Protein 1 (HMGB1) has been identified as a pivotal mediator of inflammation. HMGB1 can be released by various mechanisms and as an inflammatory mediator it induces both migration of inflammatory cells and cytokine production. Consequently, HMGB1 has been demonstrated to contribute to pathology in several inflammatory conditions. Increasing evidence indicate that HMGB1 post-translational modifications (PTMs) regulate both secretion and function of HMGB1. The focus of this thesis work has been to investigate how selected PTMs regulate HMGB1 function and release and to define the presence of such modifications on HMGB1 in synovial fluid from patients with juvenile idiopathic arthritis (JIA). Furthermore, I have set the basis for HMGB1-blockade as a clinical treatment option by generating and characterizing the first known chimeric, humanized anti-HMGB1 antibody. To examine the impact of redox-dependent PTMs on HMGB1 function, we first generated several cysteine redox isoforms of HMGB1. We found that all cysteines residues (C23, C45 and C106) required a defined redox state. A disulfide bridge between C23 and C45 with a concomitant C106 thiol was necessary for HMGB1 mediated cytokine-induction. In this disulfide redox isoform, HMGB1 activates TLR4. Furthermore, I have studied PTMs and their impact on HMGB1 secretion. We demonstrated that NLRC4 inflammasome activation induces hyperacetylation of key lysine stretches known to be associated with HMGB1 secretion, independently of priming signals. Addition of a priming signal induced reactive oxygen species (ROS) that stimulated a structural transition of HMGB1 to its cytokine-inducing, disulfide form. Hyperacetylated HMGB1 correlated significantly with inflammatory HMGB1 redox isoforms in joint fluid from JIA patients, indicating that HMGB1 is actively secreted during JIA and possesses inflammatory properties. In addition, I recorded beneficial effects of mouse monoclonal anti-HMGB1 antibody (m2G7) treatment in experimental arthritis and in acetaminopheninduced liver injury. Importantly, I could demonstrate that a partly humanized version of the antibody (h2G7) retained its in vitro properties and in vivo therapeutic effects. In conclusion, this thesis has significantly increased the understanding of the regulation of HMGB1 secretion and function during inflammation. The generation of an anti-HMGB1 chimeric antibody is an important step in development of a clinical anti-HMGB1 treatment

Roadmap for teleoperation and automation of forwarding

Annually, about two billion m3 of industrial roundwood is harvested worldwide. For a mature forest industry like the Swedish, the next technological development leaps regarding harvest operations are likely to involve increased level of tele-operation and automation. The overall aim of this thesis was to simulate and evaluate combinations of teleoperation and automation of forwarding within the cut-to-length (CTL) harvesting method, and to discuss their potential application internationally. The economic potential of different configurations of a novel concept called semiautomated tele-extraction or just tele-extraction was investigated. To evaluate the potential application of such a concept in other countries, a worldwide mapping of harvesting methods and reasons behind differences, as well as a global analysis of the slope of forest land was done. The overall result showed that there is potential for between 6% and 19% cost reduction in Swedish CTL forwarding if teleextraction is applied. Automation of more time consuming work elements, such as loading, gave larger potential than automation of driving since more operator time was deliberated. The worldwide share of fully mechanized CTL harvesting was estimated to 37% and some of its drivers include high diesel price, gross domestic product and low share of steep terrain. About 80% of global forest land had slope less than 15°, with Africa, Russia, and the Amazon rainforest especially flat. Furthermore, there is potential for implementation of tele-extraction in at least ten countries with a high share of mechanized CTL harvesting, the potential annual volume is roughly between 100 and 150 million m3

Vinster av konkurrensutsättning av den offentliga sektorn

Entreprenörskap är ett begrepp som oftast används för aktörer inom den helt privata delen av vår ekonomi. I denna uppsats utvidgas begreppet en aning. Det finns en gråzon där verksamheten visserligen finansieras kollektivt, men där privata aktörer samtidigt konkurrerar om att erbjuda konsumenterna sina tjänster. I den här uppsatsen görs försök att uppskatta eventuella vinster av att använda konkurrens som medel för att stimulera fram förbättringar inom offentlig sektor. Det kan konstateras att det föreligger en stor förbättringspotential. Mellan 25 och 50 miljarder kronor kan på kort sikt frigöras, på lång sikt antagligen betydligt mer. Men det är samtidigt av stor vikt att det regelverk som de privata entreprenörerna verkar inom utformas med omsorg så att snedvridande incitament inte uppstår.offentlig sektor; konkurrens; effektivitet; entreprenörskap

FAM13A and POM121C are candidate genes for fasting insulin: functional follow-up analysis of a genome-wide association study

Aims/hypothesis: By genome-wide association meta-analysis, 17 genetic loci associated with fasting serum insulin (FSI), a marker of systemic insulin resistance, have been identified. To define potential culprit genes in these loci, in a cross-sectional study we analysed white adipose tissue (WAT) expression of 120 genes in these loci in relation to systemic and adipose tissue variables, and functionally evaluated genes demonstrating genotype-specific expression in WAT (eQTLs). Methods: Abdominal subcutaneous adipose tissue biopsies were obtained from 114 women. Basal lipolytic activity was measured as glycerol release from adipose tissue explants. Adipocytes were isolated and insulin-stimulated incorporation of radiolabelled glucose into lipids was used to quantify adipocyte insulin sensitivity. Small interfering RNA-mediated knockout in human mesenchymal stem cells was used for functional evaluation of genes. Results: Adipose expression of 48 of the studied candidate genes associated significantly with FSI, whereas expression of 24, 17 and 2 genes, respectively, associated with adipocyte insulin sensitivity, lipolysis and/or WAT morphology (i.e. fat cell size relative to total body fat mass). Four genetic loci contained eQTLs. In one chromosome 4 locus (rs3822072), the FSI-increasing allele associated with lower FAM13A expression and FAM13A expression associated with a beneficial metabolic profile including decreased WAT lipolysis (regression coefficient, R = −0.50, p = 5.6 × 10−7). Knockdown of FAM13A increased lipolysis by ~1.5- fold and the expression of LIPE (encoding hormone-sensitive lipase, a rate-limiting enzyme in lipolysis). At the chromosome 7 locus (rs1167800), the FSI-increasing allele associated with lower POM121C expression. Consistent with an insulin-sensitising function, POM121C expression associated with systemic insulin sensitivity (R = −0.22, p = 2.0 × 10−2), adipocyte insulin sensitivity (R = 0.28, p = 3.4 × 10−3) and adipose hyperplasia (R = −0.29, p = 2.6 × 10−2). POM121C knockdown decreased expression of all adipocyte-specific markers by 25–50%, suggesting that POM121C is necessary for adipogenesis. Conclusions/interpretation: Gene expression and adipocyte functional studies support the notion that FAM13A and POM121C control adipocyte lipolysis and adipogenesis, respectively, and might thereby be involved in genetic control of systemic insulin sensitivity

Ryanodine receptors: physiological function and deregulation in Alzheimer disease

BackgroundHealth-care professionals have a responsibility to be attentive to patients’ adherence behavior but it could be difficult to identify poor adherence in the context of clinical practice. Assessment of personality could be used to identify individuals who are in need for support with their adherence behavior. To our knowledge, existing adherence questionnaires are not based on individuals reflecting asthmatics in the general population and there is limited research describing adherence with asthma medication in relation to personal goals with the treatment. The aim was to develop and validate an adherence questionnaire in adult individuals with asthma from the general population and to assess adherence in relation to personality traits and goals with the asthma medication using the developed questionnaire.MethodsThe study was conducted in three phases: 1. A preliminary postal 46-item questionnaire was refined after psychometric testing (n = 157). 2. The questionnaire was validated (n = 104). 3. The developed adherence questionnaire was analyzed in relation to personality traits and achieved goals with the asthma medication. Adult respondents with physician diagnosed asthma using asthma medications were selected from the population-based West Sweden Asthma Study. The respondents completed the Neuroticism, Extraversion and Openness to Experience Five-Factor Inventory and the Medication Adherence Report Scale and stated their goals with the asthma medication. Data were analyzed using t-tests, correlations, multiple regression and principal component analysis.ResultsA final questionnaire was developed consisting of ten items organized in three subscales - “medication routines”, “self-adjusting the medication” and “concerns about side-effects”. Two of the subscales - “medication routines” and “self-adjusting the medication” – were associated with the Medication Adherence Report Scale. The subscale “medication routines” was associated with the personality traits – Conscientiousness and Neuroticism and unachieved goals with the asthma medication.ConclusionsThe developed questionnaire appears to be useful for measuring adherence to asthma medication in adult individuals with asthma. The study suggests that both individual differences and personal treatment goals need to be addressed in efforts to promote adherence to asthma medication treatment

Stem growth, biomass production and carbon dioxide sequestration in Norrbotten after artificially and biologically fertilized Scots pine (Pinus sylvestris) forest

Denna studie är en del av ett storskaligt försök som är beläget på Sveaskogs marker i Norrbotten. Hela försöket omfattar ca 800 ha varav 600 ha är gödslad yta och 200 ha är ogödslade kontrollområden. Denna studie omfattar 137 ha gödslad yta och 50 ha ogödslade kontrollområden beläget i tre olika bestånd. Det unika med detta projekt är storskaligheten i användandet av slampellets och slamgranuler som ett förädlat bionäringsmedel. Projektet är finansierat av bland andra Sveaskog, LKAB och SLU. Tidigare studier visar på en långsiktig effekt vid gödsling med bionäring, medan denna studie undersöker bionäringens tillväxteffekt redan efter två år. Förhoppningarna med detta projekt är att sambandet mellan gödsling (mineral- och biogödsling) och ökad tillväxt samt uppbindning av CO2 (koldioxid) ska säkerställas, detta för att kunna öka nyttan av den boreala skogen som kolsänka. Genom att fastställa detta samband möjliggörs utvecklingen av ett handelssystem för uppbindningsrätter som kan kopplas samman med det befintliga handelssystemet med utsläppsrätter. Principen med uppbindningsrätterna bygger på att skogsägaren får betalt för åtgärder som skapar en ökad tillväxt och uppbindning av CO2. Gödslingen till denna studie genomfördes 2006. Varje bestånd är uppdelat i tre olika områden. Ett område är ogödslad kontroll, ett är mineralgödslat med en giva om 150kg N/ha och det sista området är gödslat med två olika givor bionäring bio a 540 kg N/ha och bio b 710kg N/ha. För att fastställa tillväxten för de olika behandlingarna klavades totalt 236 provytor 2007 och sedan igen 2009. Studiens resultat är bland annat följande: • Samtliga gödslingsbehandlingar har ökat tillväxten och uppbindningen av CO2 med minst 60 % och ända upp till 200 %. • Under försöksperioden på två år har tillväxten ökat mest på de mineralgödslade områdena. • Behandlingen bio a har ökat uppbindningen av CO2 med mellan 8,1 och 10,5 ton/ha under två år. Följande slutsatser har dragits av resultaten: • Gödsling ökar tillväxten vilket ökar CO2-bindningen. • Under en tid av två år är mineralgödsling effektivare än gödsling med bionäring • Gödsling av yngre tallskog ökar koldioxidbindningen med ca 10 ton/ha. • Gödslingen med bionäring ger en tydlig tillväxteffekt redan inom 2 år. • Högst tillväxt och koldioxidbindning sker i välsluten medelåldersskog.This study is part of a larger research project concerning carbon dioxide sequestration in northern Sweden. The testing grounds are situated within Sveaskogs properties in Norrbotten. The entire project comprises 800 hectares of which approximately 600 hectares are fertilized plots and 200 hectares are unfertilized control plots. This study comprises 137 hectares fertilized plots and 50 hectares of unfertilized control plots at three different locations. The project is unique as it concerns the use of pelletized sewage sludge as a refined biological fertilizer in a very large scale. The project is financed by Sveaskog, LKAB and SLU among others. Previous studies show that fertilization with biological fertilizers have a long term effect on forest growth. This study examines the growth response after as short a period of time as two years. It is expected to establish a relationship between fertilization (both artificial and biological) and carbon dioxide sequestration. This will enable the increase of the use of boreal forest as a carbon sink. By establishing this relationship the development of a trading scheme for carbon dioxide sequestration that can be integrated with the existing trading scheme for emitting allowances. The new trading scheme awards credits to forest owners who, with the aid of fertilization, manages their properties with a resulting increase in forest growth and therefore higher carbon dioxide sequestration. On the basis of this study the following results have been concluded: • All fertilization treatments have increased the carbon dioxide sequestration by at least 60 % and up to 200 %. • During the two year trial period the highest increase in forest growth has been observed on the plots treated with an artificial fertilizer. • The treatment Bio A increased the carbon dioxide sequestration with between 8.1 and 10.5 tons. On the basis of this study’s results the following conclusions have been drawn: • Fertilization increases forest growth and consequently carbon dioxide sequestration. • Artifical fertilizer is more effective than biological fertilizer for at least the two years directly following fertilization. • Fertilization of younger pine forest increases carbon dioxide sequestration with approximately 10 ton/hectare. • Fertilization with a biological fertilizer shows a clear growth response already within two years. • Highest growth response and increased carbon dioxide sequestration occurs in stands with larger volume

Global analysis of the slope of forest land

Forests of the world constitute one-third of the total land area and are critical for e.g. carbon balance, biodiversity, water supply and as source for bio-based products. Although the terrain within forest land has a great impact on accessibility, there is a lack of knowledge about the distribution of its variation in slope. The aim was to address that knowledge gap and create a globally consistent dataset of the distribution and area of forest land within different slope classes. A Geographic Information System (GIS) analysis was performed using the open-source QGIS, GDAL and R software. The core of the analysis was a digital elevation model and a forest cover mask, both with a final resolution of 90 m. The total forest area according to the forest mask was 4.15 billion hectares whereof 82 per cent was on slope < 15°. The remaining 18 per cent was distributed over the following slope classes, with 6 per cent on a 15–20° slope, 8 per cent on a 20–30° slope and 4 per cent on a slope > 30°. Out of the major forestry countries, China had the largest proportion of forest steeper than 15° followed by Chile and India. A sensitivity analysis with 20 m resolution resulted in increased steep areas by 1 per cent point in flat Sweden and by 11 per cent points in steep Austria. In addition to country-specific and aggregated results of slope distribution and forest area, a global raster dataset is also made freely available to cover user-specific areas that are not necessarily demarcated by country borders. Apart from predicting the regional possibilities for different harvesting equipment, which was the original idea behind this study, the results can be used to relate geographical forest variables to slope. The results could also be used in strategic forest fire fighting and large-scale planning of forest conservation and management

The role of thyroid stimulating hormone receptor and novel candidate genes FAM13a and POM121c in adipocyte dysfunction

This thesis aimed to investigate the role of thyroid stimulating hormone receptors (TSHRs) in adipose tissue (AT) and their role in AT dysfunction and obesity-related metabolic complications. Furthermore, we aimed to functionally evaluate novel candidate genes associated with insulin resistance, a marker of dysfunctional adipose tissue. Study I report that in a cohort of Swedish children with obesity, thyroid stimulating hormone was associated with the degree of obesity and metabolic risk markers such as fasting serum insulin levels and blood lipids. In Study II, the TSHR in AT was functionally evaluated by investigating the effect of partially removing TSHRs from the adipocytes on body weight and body temperature, in animals on both normal and high-fat diets. Mice with reduced TSHR-expression gained weight at a faster rate than corresponding wild-type mice. Several genes central to adipogenesis and adipocyte function were down-regulated, in both white AT (WAT) and brown AT (BAT) in TSHR knockout mice. Study III. From a genome-wide association study (GWAS) meta-analysis which identified SNPs for fasting insulin, we identified FAM13A and POM121C as novel candidate genes for obesity-related insulin resistance. Using expression quantitative trait (eQTL) analysis of SNPs associated with fasting insulin, we identified candidate genes for disease. Functional analysis of the candidate genes using siRNA knockdown in human mesenchymal stem cells revealed them to be important for lipolysis and adipogenesis and they might therefore be involved in the genetic control of insulin sensitivity. Study IV, report that expression of the TSHR in human WAT is affected by weight change. We also report that TSHR-expression in WAT is associated with the expression of genes central to adipocyte functions such as lipolysis and insulin sensitivity. The results revealed that, independent of BMI, individuals with higher TSHR expression had a lower basal lipolysis rate and higher hormone stimulated lipolysis, suggesting that TSHRs in human WAT are involved in the regulation of adipocyte metabolism. In summary, a reduction of TSHR led to dysfunctional AT regarding the regulation of adipocyte metabolism and adipogenesis. Our findings implicate that TSHRs have a regulatory role in both WAT and BAT, and thus having a role in the regulation of whole-body energy homeostasis. Furthermore, we identified two novel genes with potential regulatory roles in adipocyte lipolysis and adipogenesis, and might thereby be involved in the genetic control of systemic insulin sensitivity