Idag står världen inför nya typer av utmaningar för att möta hållbarhetsmål och nya behov somuppkommer. Många städer genomgår därför stora förändringar i utvecklingen där både förväntadeoch oväntade konsekvenser kommer att uppstå. Sedan Northvolt beslutade att etablera sig iSkellefteå år 2017, står staden inför den stora möjligheten att bli en pionjär stad för grön utveckling.Den nya utvecklingen har inte bara satt industrin på prov, utan även staden som helhet. Kommunensutmaning är att sprida budskapet om vad den kommande utvecklingen har att erbjuda, i hopp om attsätta fart på inflyttningen då kompetensbehovet är Skellefteås främsta utmaning idag. För att mötavisionerna kring utvecklingen behöver alla aktörer samverka för att bli en stad för grön elektrifiering.Välfungerande socialt kapital mellan offentlig sektor, industri, akademi och medborgare är enförutsättning för att skapa en framgångsrik utveckling.Arbetet utforskar hur en stad som genomgår en omfattande och snabb industriell expansionpåverkas. Genom att studera migrationsmönster ur “push and pull”-teorin undersöker arbetet hur detär möjligt att generera en ökad in-migration i olika målgrupper, som studenter eller hemvändare.Samtidigt som Skellefteå är beroende av att ha ett positiv inflyttningsnetto behöver även denuvarande invånarna bibehållas. Därför har intervjuer och en enkät genomförts för att få en tydligbild av styrkor och svagheter i utvecklingen av Skellefteå. Resultaten av enkäten och intervjuernaanalyserades och diskuterades genom en exemplifiering av de två teorierna push and pull och teorinom socialt kapital.Stora industriella expansioner av denna storlek skakar städerna till sin kärna, eftersom stadensidentitet och attribut ifrågasätts under denna typ av förändring. Vid en utveckling av denna storleksätts det sociala kapitalet på prov och beroende på dess effektivitet kan utfallen skilja sig åt. Stadentvingas väga olika faktorer mot varandra; ekonomiska, sociala och miljömässiga, då alla inte är möjligaatt upprätthålla till samma grad. Eftersom det största målet är att skapa en ökad in-migration till denexpanderande staden måste de pull-faktorer som finns och främjas ifrågasättas; Vad erbjuderegentligen Skellefteå? Arbetet identifierade sex möjliga grupper av in-migration; kontraurbanister,livsstilsmigranter, människor som flyttar för relationer/hemvändare, människor som flyttar förjobbmöjligheter, studenter och människor som flyttar för hållbarhetsskäl. Genom att lyfta fram enuppsättning av pull-faktorer specifikt för Skellefteå har staden den unika möjligheten att påverkamigrationsmönstren till deras behov och skapa en hållbar och attraktiv stad för den nuvarande ochframtida invånaren. / Today the world stands in front of the challenge of adapting to new demands, where solutions thatincrease sustainability are required. Many cities are therefore undergoing big changes indevelopments where both expected and unexpected consequences will arise. Since 2017, whenNorthvolt decided to establish in Skellefteå, the city stands in front of the opportunity to become apioneer city of green evolution. The new development has not only put the demands of the industryat test, but also the city as whole. The challenge of the municipality is how to spread the message ofwhat the upcoming development has to offer to boost the in-migration, as the need for competence isthe main challenge of Skellefteå today. To be able to fulfil and meet the visions of the transformation,all actors need to work together in becoming the green city of electrification. To create a successfuldevelopment, the overlapping collaboration between the government, industry, academia andcommunity needs to be of high quality and thus the social capital will increase.The thesis explores the effects of a city that undergoes a fast expansion. By studying migrationpatterns through push and pull theory, the thesis investigates how to generate an increasedin-migration in different target groups, such as students and returners. While Skellefteå is dependenton having a positive netto in-migration, the current residents also need to be maintained, thusinterviews and a questionnaire has allowed the thesis to obtain a clear picture of strengths andweaknesses of the development in Skellefteå. The results of the questionnaire and the interviews areanalysed and discussed through an exemplification of the two theories of push and pull and thetheory of social capital.Large industrial expansions of this size shake cities to their core, as the identity and attributes of thecity are questioned as they change. When dealing with a development of this size, the social capital isput to the test and depending on its effectiveness, the outcomes can differ. The city is forced to weighdifferent factors against each other, the benefits of accommodating economic, social andenvironmental factors are measured and prioritised as all are not possible to sustain. As the main goalis to create an increased in-migration to the expanding city, the pull factors available and promotedneed to be questioned; What is Skellefteå in fact offering? The thesis identified six possible groups ofin-migration; counterurbanists, lifestyle migrants, people moving for social ties/the returners, peoplemoving for job opportunities, students and people moving for sustainability reasons. By highlightingone set of pull factors, Skellefteå has the unique opportunity to influence the migration patterns totheir needs and create a sustainable and attractive city for the current and future residents.

The next generation of toxic chemicals and hazardous wastes from sophisticated chemical industries will demand the environmental agencies to employ biological methods over the conventional physical and chemical remediation methods. Over the past decade, natural metallo-enzymes have been identified to degrade some of the major chemical contaminants through electron transfer pathways. However, these natural enzymes are less stable in organic solvents and they are not effective for the degradation of toxic compounds such as polychlorinated biphenyls or dioxins. This thesis explores the use of protein design approaches to produce chemically and molecularly modified enzymes, which are highly stable, possess little substrate specificity, and have higher activity than the natural enzymes. The experiments presented in this thesis make use of solid phase synthesis and site-directed mutagenesis for the synthesis and production of these enzymes and popular chromatographic techniques for their purification. The partial characterization of these proteins revealed the essential structural features of these proteins, and their catalytic activity was demonstrated by the use of peroxidase assays. / Master of Science

The principal objective of this project was to determine whether Hedera helix was distributed randomly or exhibited host preferences within Drayton Wood, Norwich. 200 random tree samples >10cm diameter were studied. Of these, the CBH (circumference breast height), bark texture, abundance of H. helix and growth form were recorded. The percentage of ground covered by H. helix around the base of the sample trees was also recorded. Finally, the distance of the sample trees from the outer reaches of the wood and canopy cover were measured.Hedera helix was not distributed over potential host trees homogenously, but exhibited host associations. Quercus robur, Crataegus monogyna and Pinus sylvestris tree species were found to have a higher abundance of H. helix than expected by chance, whereas Acer pseudoplatanus and Betula pendula were found to have less H. helix vines than expected. Furthermore juvenile and mature vines were not distributed equally. Acer pseudoplatanus hosted more juveniles and less adults than expected, whereas Quercus robur and Pinus sylvestris hosted less juveniles and more mature vines than expected. Vines were not distributed independently of host size. Those trees <100cm CBH had a lesser stem abundance than expected, whereas those vines >100cm CBH had a greater stem abundance than expected. Host preference was exhibited for those large CBH trees with rough bark as opposed to large CBH trees with smooth bark. There was a higher abundance of H. helix (ground and trunk cover) with distance from the edge. Ground cover was greater under those trees hosting H. helix than those not supporting H. helix.Light did not play a major role in the distribution and abundance of H. helix (ground cover and trunk cover).Key Words: host selectivity, host size, bark texture, ground cover, location, light.

No / Developmental periods in early life may be particularly vulnerable to impacts of environmental exposures. Human research on this topic has generally focused on single exposure–health effect relationships. The “exposome” concept encompasses the totality of exposures from conception onward, complementing the genome. Objectives: The Human Early-Life Exposome (HELIX) project is a new collaborative research project that aims to implement novel exposure assessment and biomarker methods to characterize early-life exposure to multiple environmental factors and associate these with omics biomarkers and child health outcomes, thus characterizing the “early-life exposome.” Here we describe the general design of the project. Methods: In six existing birth cohort studies in Europe, HELIX will estimate prenatal and postnatal exposure to a broad range of chemical and physical exposures. Exposure models will be developed for the full cohorts totaling 32,000 mother–child pairs, and biomarkers will be measured in a subset of 1,200 mother–child pairs. Nested repeat-sampling panel studies (n = 150) will collect data on biomarker variability, use smartphones to assess mobility and physical activity, and perform personal exposure monitoring. Omics techniques will determine molecular profiles (metabolome, proteome, transcriptome, epigenome) associated with exposures. Statistical methods for multiple exposures will provide exposure–response estimates for fetal and child growth, obesity, neurodevelopment, and respiratory outcomes. A health impact assessment exercise will evaluate risks and benefits of combined exposures. Conclusions: HELIX is one of the first attempts to describe the early-life exposome of European populations and unravel its relation to omics markers and health in childhood. As proof of concept, it will form an important first step toward the life-course exposome.

This study addresses the need to enhance Sweden's defence capability considering global changes and Russia's invasion of Ukraine, with a focus on defence innovation. It examines how the Swedish government's assumptions about defence innovation shape defence capabilities, emphasizing the government's strategic direction for defence innovation. Using Carol Bacchi's WPR method and critical discourse analysis, the study identifies two central problem representations: inertia and isolation. Inertia refers to the slow adaptation to new technological and organizational changes, while isolation pertains to the lack of collaboration between the defence sector and civilian actors. These representations lead to a discourse advocating increased risk-taking and faster development, often overlooking potential negative consequences. The study highlights the need for a balanced strategy that integrates innovation and conservatism to ensure sustainable defence development. Understanding these assumptions can help develop a more holistic and long-term strategy, which is crucial for strengthening Sweden's defence capability.

Transcription factors of the helix-loop-helix (HLH) gene family are widespread in the animal kingdom. Among them, members of HLH subfamilies such as ASCL, Neurogenin, NeuroD, COE, Atonal, Oligo, NSCL, Hairy/E(spl) and Hey (here referred to as neural HLH genes) have been shown to be fundamental for the development of the nervous system. They are expressed at different time periods of neuronal differentiation, from the specification of ectoderm towards a neural lineage, to the ultimate differentiation of neurons. Few HLH genes have been identified in the lamprey; however, considering the wide diversity of HLH gene subfamilies in metazoans, including vertebrates, it is very likely that lampreys possess a large repertoire of HLH genes in their genome. In the present study, the identification of several HLH genes in the lamprey genome, as well as the isolation and expression of different lamprey neural HLH genes is reported. As expected, a wide repertoire of HLH genes was identified in the sea lamprey (Petromyzon marinus) genome. On the other hand, the identification and expression analysis of different neural HLH genes of the ASCL, Neurogenin, COE and Hairy/E(spl) in the brook lamprey Lampetra planeri showed an overall conservation with other vertebrates, both at the sequence and expression pattern levels. In addition, novel features of the lamprey nervous system are revealed, such as the identification of possible new sensory cranial placodes in pharyngeal arches. Furthermore, these genes can serve as molecular markers for different cranial placodes and dorsal root ganglia (DRG), and their expression also highlights the presence of a ventricular zone in the brain and spinal cord, along with a complementary marginal zone. Finally, with the use of a Notch pathway inhibitor in developing L. planeri embryos, the regulation of expression of the isolated genes by the Notch signaling pathway was shown to be generally conserved between lampreys and gnathostomes in the spinal cord. This functional study also revealed that the lamprey spinal cord likely presents an independent developmental programme from the brain. All together, the present study shows that the analysis of neural HLH genes represents an excellent tool to understand the lamprey nervous system.

The Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor and member of the basic helix-loop-helix Per/ARNT/Sim (bHLH/PAS) family of chemosensors and developmental regulators. As a member of the PAS domain family of transcription factors responsive to exogenous signals, the AhR exerts influence on many processes relating to cellular fate. The activation of AhR is widely associated with toxic endpoints related to dioxin exposure. However, the AhR also activates endogenous gene programs related to development, cellular growth, and differentiation. The AhR is able to bind a variety of ligands, leading to a wide range of biological outcomes. Recent reports have shown that the AhR can mediate tumor suppressive effects. As a ligand-activated transcription factor, the AhR has the potential to actuate a variety of transcriptional programs that are dependent on the AhR ligand. Our central hypothesis is that AhR ligands can be identified that are capable of initiating tumor suppressive functions of the AhR. We utilized complementary cell-based and in silico virtual screening approaches to identify potential AhR ligands. We developed homology models of the AhR ligand-binding domain (LBD) for virtual ligand screening (VLS) of small molecule libraries. This led to the identification of new AhR ligands 5,7- dihydroxyflavanone!and 5-hydroxy-7-methoxyflavone. Additional small molecule libraries were screened in parallel that led to identification of flutamide as a putative AhR ligand. Flutamide is clinically approved for the treatment of prostate cancer due to its ability to antagonize androgen receptor mediated transcription. We investigated the biological effects of flutamide in AhR positive cancer cells that do not express the androgen receptor and found that flutamide inhibited the growth of HepG2 cells. Suppression of AhR expression reversed the anti-proliferative effects of flutamide. We tested 15 structural analogs of flutamide, including the flutamide metabolite 2-hydroxyflutamide for activation of AhR transcriptional activity. Flutamide is unique in its ability to activate the AhR, and suppresses hepatoma cell growth. These data suggests that flutamide-induced AhR transcriptional activity is required to initiate the tumor suppressive effects. We examined changes in cell cycle checkpoint proteins after flutamide treatment and discovered increased expression of cell cycle inhibitory proteins p27[superscript Kip1] and p15[superscript INK]. We also found that transforming Growth Factor β1 (TGFβ1), which regulates both p27[superscript Kip1] and p15[superscript INK], is upregulated by flutamide. We demonstrate that TGFβ1 is upregulated by flutamide in an AhR-dependent manner and is required for suppression of proliferation by flutamide. We identify specific and unique transcriptional signatures of the AhR upon activation by flutamide, that are distinct from the potent AhR agonist 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD). In summary, we characterize flutamide as an AhR ligand and demonstrate its AhR-dependent tumor suppressive effects in hepatoma cells. We provide the first direct evidence that AhR regulates TGFβ signaling in a ligand dependent manner. We demonstrate that the AhR-induced downstream transcriptional signature and subsequent biological effects are specific to the AhR ligand. Our studies have broad impact for characterizing the AhR as a new therapeutic target in hepatocellular carcinoma. / Graduation date: 2013 / Access restricted to the OSU Community at author's request from March 28, 2013 - March 28, 2015

HIV-1 has evolved with a great deal of functional complexity contained within a very small genome by encoding small, but critical viral proteins within larger viral genes and dividing the replication cycle into early and late phases to differentially produce all proteins leading to efficient replication and virion release. Early replication is restricted by the host spliceosome that processes HIV-1 vRNA transcripts so only the small intragenomic proteins are produced, one of which is Rev (Regulator of Virion Expression). Rev in turn governs the transition from early to late replication by interacting with a highly structured region of vRNA termed the Rev Response Element (RRE). The binding of Rev to the RRE is believed to cause a change in the vRNA tertiary structure and inhibition of splicing of the vRNA. Once, a Rev:RRE complex is formed, a nuclear export signal within Rev facilitates the export of partially spliced and unspliced vRNA to the cytoplasm. During late replication the partially spliced and unspliced vRNA is translated to polyproteins and is packaged into a budding virion where the viral aspartyl protease (HIV-1 PR) autocatalytically excises itself from the larger polyprotein and processes the remaining polyproteins to release all viral structural and functional proteins to form a mature and infectious virion. Since the vRNA salvaged by Rev is translated to the polyproteins containing HIV-1 PR, the inhibition of Rev function will reduce the amount of HIV-1 PR available and thereby reduce the amount of HIV-1 PR therapeutics required to elicit a clinical effect. Therfore a combination approach to HIV-1 treatment using suitably developed therapeutics that inhibit Rev and HIV-1 PR function represents an attractive synergistic approach to treating HIV-1 infection in vivo. The work of this thesis was divided into two parts, the first part was concerned with HIV-1 PR structural biology and addressing problems encountered with inhibitor design. A bicyclic peptide (based on inhibitors of analogous structure) was co-crystallised with active HIV-1 PR to develop an enzyme-product (E-P) complex and with a catalytically inactive mutant HIV-1 PR to provide an analogy to the enzyme-substrate (E-S) complex. Both structures of the E-P and E-S complexes were solved to 1.6Å resolution and were compared to a hydroxyethylamine isostere enzyme-inhibitor complex (E-I), highlighting the similarity of binding mode for all ligands. The inhibitor in the E-I complex was translated towards the S1 - S3 pockets of the substrate binding cleft relative to the substrate in the E-S complex due to the increased length of the hydroxylethylamine isostere compared to the peptide backbone, although the inhibitor "puckered" the isostere linkage and maintains a binding mode similar to the substrate with very little overall differences in the position of the ligands and surrounding protein. The similarity of the E-S, E-I and E-P complexes was attributed to the macrocyclic ligands ordering the surrounding protein environment, especially the protein -strand "flap" structures that form a roof over the ligands in the active site but were not found to close more tightly in any of the trapped catalytic states. The new structures allowed refinement of details of the mechanism of peptide hydrolysis. The mechanism relies on the optimal nucleophilic attack of a water molecule on the scissile amide bond with concerted acid-base catalysis of the active site aspartyl residues intitiated by D125. The alignment and intrinsic position of the N-terminus of the bicyclic substrate was interpreted as being critical to facilitate efficient electron transfer with the bicyclic substrate. An N-terminal cyclic inhibitor, similar to the N-terminal portion of the bicyclic substrate, was used to address a major problem in HIV-1 PR drug design termed "cooperativity," where the sequential optimisation of an inhibitor (or substrate) to individual pockets of the substrate binding cleft, can negatively impact on adjacent and downfield subsites and thereby alter the binding mode of the "optimised" inhibitor. The technique referred to here as "templating" uses the N-terminal cycle to lock the binding mode into a known conformation, probing the S1' and S2' pockets. The structure activity relationship suggested that by viewing the S1' - S3' pockets as a single trough, bulky aromatic groups attached to an N-alkyl sulfonamide could be directed along the line of the trough without adverse interactions with the tops of the S1' and S3' pockets, providing very potent inhibitors. It was also found that specificity and potency of an inhibitor can be maintained with smaller functionalities that carry their bulk low and close to the inhibitor backbone in the S2' pocket, making the P2 functionalities more substrate-like. The second part of the thesis was concerned with establishing suitable surface plasmon resonance assays for testing potential inhibitors of Rev function. Recombinant Rev and its minimal RNA aptamer target (stem loop II of the RRE termed RBE3), were expressed, purified, and used to develop BIAcore-based assays and test potential inhibitors of their interaction. The system was applied to screening of aminoglycoside antibiotics and other small molecules in a competitive assay, and also to quantitative assay of Neomycin and moderate sized analytes: Rev and three peptidic analogues of the high-affinity binding site of Rev - the native peptide, succinylated form of the peptide and a form incorporating a novel helix-inducing cap. The peptide and protein assay was undertaken to test the proposition that helix induction of the high-affinity binding site of Rev can increase affinity for the biologically important RNA target and thereby form the basis of a new class of inhibitors. The screen of small molecule antagonists found that Neomycin was the best inhibitor of the Rev:RBE3 interaction and that efficacy of other aminoglycosides was due to the neamine-base structure presenting charge to bind to the RNA and blocking interaction with Rev. The quantitative assay was optimised to reduce non-specific interactions of Rev protein to allow reliable studies of the analytes with RBE3 by the sytematic testing of buffers and modifiers. It was found that mutliple analytes bound to the RBE3 aptamer and a comparison of the KD values found that the native and capped peptides had similar affinity for RBE3 RNA (native slightly greater at 21 ± 7nM cf capped 41 ± 10nM) that was greater than the Rev protein (101 ± 19nM), however the succinylated peptide exhibited stronger binding with a KD ≤8nM and Neomycin had the lowest affinity (KD 13 ± 3M). The similarity of the native and capped peptides may be due to the high concentration of salt in the assay buffers and was necessary for the stability of the Rev protein, but is sufficient to influence secondary structure of the peptides. Therefore, it could not be stated that the helix-inducing cap increased the affinity of the native peptide for the biologically important therapeutic target. The work conducted in this thesis firmly establishes foundations for the continued development of inhibitors against both Rev and HIV-1 PR that play key roles in the HIV-1 replication strategy. It is envisaged this work could lead to a novel synergistic therapeutic approach to treating HIV-1 infection.

Diss. (sammanfattning) Umeå : Umeå universitet, 2008. / Härtill 5 uppsatser.

Activation of the basic helix-loop-helix (bHLH) gene TAL1 is the most common genetic event seen in both childhood and adult T cell acute lymphoblastic leukemia (T-ALL). Despite recent success in treating T-ALL patients, TAL1 patients do not respond well to current therapies. In hopes of leading the way to better therapies for these patients, we have sought to determine the mechanism(s) of Tal1 induced leukemia in mice. By generating a DNA-binding mutant Tal1 transgenic mouse we have determined that the DNA binding activity of Tal1 is not required to induce leukemia. We have also shown that Tal1 expression in the thymus affects thymocyte development and survival. We demonstrate that Tal1 heterodimerizes with the class I bHLH proteins E47 and HEB in our mouse models of TAL1 induced leukemia. Severe thymocyte differentiation arrest and disease acceleration in Tal1/E2A+/- and Tal1/HEB+/- mice provides genetic evidence that Tal1 causes leukemia by inhibiting the function of the transcriptional activators E47 and HEB which have been previously shown to be important in T cell development. In pre-leukemic Tal1 thymocytes, we find the co-repressor mSin3A/HDAC1 bound to the CD4 enhancer, whereas an E47/HEB/p300 complex is detected in wild type thymocytes. Furthermore, mouse Tal1 tumors are sensitive to pharmacologic inhibition of HDAC and undergo apoptosis. These data demonstrate that Tal1 induces T cell leukemia by repressing the transcriptional activity of E47/HEB and suggests that HDAC inhibitors may prove efficacious in T-ALL patients that express TAL1.