Syftet med uppsatsen är att synliggöra underliggande ekokritiska teman i Nils Holgerssons underbara resa genom Sverige och att undersöka människans relation till naturen med hjälp av verket. Uppsatsen använder sig av teori från ekokritik och djurstudier och undersöker frågeställningar grundade efter dessa fält, nämligen: hur gestaltas naturen i Nils Holgersson, hur gestaltas människorna i Nils Holgersson, hur gestaltas djuren i Nils Holgersson och hur gestaltas relationerna mellan människa, djur och natur? För att besvara syftet och frågeställningarna utför jag en tematisk läsning av verket och analyserar den med hjälp av ekokritiska begrepp och termer samt tidigare forskning kopplat till både Nils Holgerssons underbara resa genom Sverige och ekokritik. Mina analyser tyder på att även om geografin skulle vara central i boken så har Lagerlöfs val att inkludera moralhistoria satt sina spår i gestaltningen av människa, djur och natur till en mer antropocentrisk världsbild. Naturen i Nils Holgerssons underbara resa genom Sverige beskrivs oftast med ett instrumentellt värde, det vill säga antropocentriskt. Människorna gestaltas som en konstant motpol till och i konflikt med djuren, men framhålls i slutändan som något åtråvärt för Nils Holgersson. Djuren beskrivs oftast med mänskliga drag där gränsen mellan människa och djur oftast suddas ut, och de beskrivs oftast med antropomorfiska drag, likt fabler. Relationen mellan människa, djur och natur är konfliktfylld där såväl djur som människa försöker utnyttja naturen, men där människan alltid har det minst ekocentriskt överstämmande handlingsättet. Nils Holgersson som karaktär kan till stor del beskrivas som antropocentrisk men verket som helhet kan i slutändan ses somen lära i ekocentrism, och därmed har verket en fortsatt viktig roll för forskning inom ekokritik ochdjurstudier / <p></p><p></p>

Denna studie utforskar hur vi kan förhålla oss till naturliga material och lokala resurser i samband med att färgtillverkning inkorporeras som en aktivitet i gymnasieskolans bildundervisning. Idag är inte färgtillverkning något som ofta görs i skolor, då mycket av det färgmaterial som används kommer färdigblandat i en tub eller kaka. Studien lyfter frågor som berör vilka möjligheter till lärande som skapas när färgtillverkning integreras i gymnasieskolans bildundervisning samt hur tillverkning av färg skulle kunna bidra till en djupare förståelse för måleri som både ett hantverk och en konstnärlig uttrycksform. Undersökningen lyfter även frågor om varför detta ämne är relevant idag, med en koppling till ett hållbarhetsperspektiv i färgtillverkningsprocesser i bildpedagogiska situationer.  Med hjälp av forskningsmetoden research-creation organiseras fyra forskningshändelser, vars syfte är att skapa en djupare förståelse för färgtillverkningsprocesser och öppna upp för hur detta skulle kunna inkorporeras i skolan.   Utöver denna uppsats har undersökningen resulterat i en visuell gestaltning i form av en film som visar en färgtillverkningsprocess som ställdes ut på Konstfacks vårutställning.  Teorier om att lära genom att göra, samt multimodal teori med hjälp av formuleringen resources and their affordances används för att skapa en tolkningsram för de olika forskningshändelserna.  Djupekologi används även som en filosofisk inriktning för att diskutera hur människan idag kan uppfattas som frånkopplad från naturen. Denna filosofiska inriktning vill lyfta hur en vändning till att utforska naturliga material och lokala resurser kan bidra till en djupare förståelse och uppskattning för den materiella världen i skapandeprocesser.

Foamy viruses (FVs), like other retroviruses, have gained interest due to their applications as viral vectors in gene therapy and various other fields. Despite their potential applications, FV stands out as a unique retrovirus, with distinct features in its replicative cycle that make it a compelling subject of study. However, it remains one of the least studied retroviruses. A crucial aspect of the retroviral replicative cycle, especially for its use as in gene transfer, is the selective packaging of its viral genomic RNA (vgRNA) out of the vast pool of RNAs found in virus producing host cells. This process, known as vgRNA enrichment, involves efficiently packaging vgRNA among the cellular and processed viral genetic material. Unlike other retroviruses, the selective packaging process of vgRNA in FV has not been explored, making it largely unknown. Previous studies on other retroviruses have indicated that vgRNA packaging selectivity is achieved through specific features within the vgRNA itself, such as specific sequences elements called packaging signals as well as dimerization of vgRNA resulting in the presence of two copies vgRNA in each retrovirus particle. Additionally, viral structural proteins, particularly Gag, play a significant role by interacting with vgRNA through specific protein regions. To shed light on the selective packaging process of FV vgRNA, the roles of structural proteins Env, Pol, and Gag were investigated, along with the vgRNA's dimerization capacity. To quantify the vgRNA selective packaging, an enrichment assay (E-assay) and a competitive assay (C-assay) were established. The E-assay allows to compare FV vgRNA packaging efficiency relative to non-viral RNAs, whereas the C-assay enables the determination of the preferential packaging of one vgRNA over another when both are present within a cell. The results obtained regarding the role of the structural proteins on the vgRNA selective packaging emphasized the delicate balance required between viral protein expression and vgRNA levels. Overexpression of Env and Gag severely disrupted selective packaging. Particularly the excess of Env protein amount led to an increased production of subviral particles that lack the capability to selectively package vgRNA. An interesting observation was the impact of the RNA template used for translating Gag on vgRNA enrichment. Expressing Gag from vgRNA (cis) enhanced vgRNA packaging selectivity, while expression from an RNA containing only expression-optimized gag ORF sequences (trans) reduced vgRNA enrichment. Nevertheless, the results from the C-assays suggest that non-Gag-translating vgRNA can still be selectively packaged over non-selectively packable dimerization deficient vgRNA. This indicates that while Gag might have a cis-acting mechanism in FV vgRNA selective packaging, this role appears to be non-essential. As mentioned earlier, vgRNA dimerization appears to be a crucial factor in the selective packaging process of most retroviruses. In the case of FV, the dimerization process was previously reported to be facilitated by three specific regions on the vgRNA known as dimerization sites one to three (DS-I to -III). Among these sites, DS-II stands out as being indispensable for vgRNA dimerization due to its 10 nt palindromic sequence, a determinant reported to be essential for the interaction between the two strands of vgRNA for most retroviruses. To investigate the significance of FV vgRNA dimerization in its selective packaging, we conducted E- and C-assays to assess the vgRNA packaging efficiency and specificity in FV DS-II mutants, previously identified as non-dimerizing (DS-II-M6 and -M7) or exhibiting a low dimerization rate (DS-II-M2). Intriguingly, FV vgRNA packaging was significantly negatively affected in the non-dimerizing FV mutants (DS-II-M6 and -M7), and to a lesser extent in the DS-II-M2 mutant, where dimerization was reported of occur at lower rates. This reveals a direct correlation between vgRNA packaging efficiency and the reported vgRNA dimerization potential of these DS-II mutants. These results suggest that, similar to other retroviruses, vgRNA dimerization plays a pivotal role in FV's selective packaging. Furthermore, it is well-documented that secondary structures within vgRNA in some retroviruses facilitate dimerization, thereby enhancing the selective packaging process. In-silico analyses of the FV vgRNA predicted the formation of a stem loop created by the palindromic sequence (SL-Pal). To gain insight into the role of these secondary structures within the DS-II region of FV vgRNA in its selective packaging, we designed a series of new dimerization mutants. These mutants were meticulously engineered to disrupt, modify, or restore the SL-Pal structure by introducing mutations inside or in proximity to the palindromic sequence based on computational secondary structure prediction. Notably, we observed that the palindrome's sequence could be mutated, as long as the SL-Pal structure and G-C proportion along all the stem-loop were preserved in a manner identical to the original structure. This preservation was crucial to ensuring the selective packaging of vgRNA and maintaining viral infectivity. Lastly, the evaluation encompassing protein analysis, vgDNA quantification, and infectivity assessment conducted on DS-I and DS-II mutants revealed a significant decrease not only on vgRNA selective packaging but also on viral infectivity, Pol packaging, cleaving, and the RTr process in non-dimerizing mutants. This underscores the intricate interrelation of these processes, emphasizing their collective importance for successful viral production. In summary, the findings presented in this project represent a significant advancement in understanding FV vgRNA selective packaging and dimerization. They offer valuable and novel insights that contribute to the expansion of our knowledge about FV molecular biology and its potential applications as a viral transfer vector.:I. ACKNOWLEDGEMENTS I II. TABLE OF CONTENT III III. INDEX OF FIGURES VI IV. INDEX OF TABLES VIII 1 INTRODUCTION 1 1.1 Retroviruses 1 1.1.1 Taxonomy of retroviruses 2 1.1.2 General features of retroviruses 3 1.2 Foamy viruses 6 1.2.1 PFV virion structure and genome organization 8 1.2.2 Viral proteins 11 1.2.2.1 Gag 11 1.2.2.2 Pol 13 1.2.2.3 Env 15 1.2.3 Replication cycle 17 1.2.3.1 Early phase 18 1.2.3.2 Late phase 19 1.3 Selective packaging of retroviral vgRNA 20 1.3.1 selective vgRNA packaging in orthoretroviruses 20 1.3.1.1 Role of the capsid protein Gag on the selective vgRNA packaging 20 1.3.1.2 vgRNA packaging signals 22 1.3.1.3 vgRNA dimerization and selective packaging 23 1.3.1.4 Factors that determine the vgRNA fate 24 1.3.2 Selective packaging in Hepadnaviruses 26 1.3.3 Selective packaging in PFV 26 2 THESIS AIM 29 3 MATERIALS AND METHODS 30 3.1 Buffers and solutions 30 3.2 Enzymes 34 3.3 Commercial kits 34 3.4 Nucleic acids 35 3.4.1 Oligonucleotides 35 3.4.1.1 Oligonucleotides for cloning 35 3.4.1.2 Oligonucleotides for qPCR analysis 37 3.4.2 Plasmids 39 3.4.2.1 Plasmid constructs used in this project: 39 3.4.2.2 New plasmid constructs 42 3.5 Bacteria strains 52 3.6 Cell lines 52 3.7 Antibodies 53 3.8 Software and Devices 54 3.9 Consumables 56 3.10 Molecular Biology methods 56 3.10.1 Bacteria culture 56 3.10.2 Transformation of competent bacteria 57 3.10.3 Plasmid extraction 57 3.10.4 Molecular cloning 58 3.10.4.1 Polymerase Chain Reaction (PCR) 58 3.10.4.2 Plasmid digest 59 3.10.4.3 Fragment purification 59 3.10.4.4 Ligation 60 3.10.4.5 Transformation of ligated plasmid constructs 60 3.10.4.6 Plasmid preparation small-scale 61 3.10.4.7 Plasmid Sequencing 61 3.10.4.8 Plasmid quantification 61 3.10.4.8.1 Photometric quantification 61 3.10.4.8.2 Fluorometric quantification 62 3.1 Biochemistry methods 62 3.1.1 SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE) 62 3.1.2 Semi-Dry Western Blot 63 3.1.3 Immunodetection 63 3.1.4 Quantification of viral proteins by Western Blot 63 3.2 Cell Culture and Virological Methods 64 3.2.1 Cell lines 64 3.2.2 Cell passaging 64 3.2.3 Calcium phosphate transfection 65 3.2.4 Harvest of viral particles 65 3.2.5 Cell harvesting 66 3.2.6 Viral infectivity determination 66 3.3 Molecular virology methods 67 3.3.1 Viral RNA extraction 67 3.3.2 Cellular RNA extraction 67 3.3.3 DNase treatment 68 3.3.4 RNA quantification by RT-qPCR 68 3.3.5 DNA quantification by qPCR 69 3.3.6 Enrichment assay (E-assay) 70 3.3.7 Competitive assay 72 3.3.8 Secondary RNA structure prediction 74 4 RESULTS 75 4.1 Establishment of methodologies for quantification of selective vgRNA packaging efficiency 75 4.1.1 Establishment of the Enrichment (E)-assay 75 4.1.1.1 Reference mRNA 75 4.1.1.2 Background subtraction 77 4.1.2 Establishment of the Competition (C)-assay 80 4.1.2.1 Silent mutants characterization 81 4.1.2.2 Primer design 83 4.2 Selective packaging in foamy virus 88 4.2.1 Role of the viral structural proteins on the selective vgRNA packaging 92 4.2.1.1 Role of Env on the selective vgRNA packaging 92 4.2.1.2 Role of Pol on the selective vgRNA packaging 97 4.2.1.3 Role of Gag on the selective vgRNA packaging 100 4.2.1.3.1 Gag cis-acting mechanism on the selective vgRNA packaging 103 4.2.2 Role of dimerization on the selective packaging 109 4.2.2.1 Characterization of the dimerization mutants 111 4.2.2.2 Selective vgRNA packaging and dimerization 114 4.2.2.3 PFV vgRNA secondary structure (prediction) and dimerization potential 118 4.2.2.4 Dimerization and RTr 124 5 DISCUSSION 129 5.1 Establishment of methodologies for vgRNA selective packaging determination 129 5.1.1 E-assay 129 5.1.2 C-assay 130 5.2 Viral factors involved in the FV vgRNA selective packaging 132 5.2.1 Selective packaging in PFV 132 5.2.2 Role of the structural viral proteins on the vgRNA selective packaging 133 5.2.2.1 Env 133 5.2.2.2 Pol 136 5.2.2.3 Gag 136 5.2.3 Role of vgRNA dimerization on its selective packaging 139 5.2.3.1 vgRNA DS-II secondary structure and dimerization 141 5.2.3.2 vgRNA dimerization and RTr 142 6 CONCLUSION 145 7 REFERENCES 146 8 APPENDICES 162 8.1 Abbreviations list 162 8.2 Anlage 1 165 8.3 Anlage 2 166

An influence of variations in human genome is perceptible at a first glance on human itself to see differences between the individuals and entire populations. Also, behavior or probability of certain diseases are influenced in large way by differences at genome's level. This work presents methods for detecting variations in the human genome that were developed after an arose of the second-generation sequencing technologies. A new tool that combines read pair and split read methods, with information about a depth of coverage was also designed and implemented. The tool was tested on simulated and real data and compared with a reference outputs.

I det här arbetet undersöks olika sätt att arbeta med fysik utifrån litteratur. Metoden har varit att intervjua förskollärare och en kontaktperson ifrån projektet Lilla bokbryggan, samt att genomföra litteraturanalyser. Resultatet visar på olika arbetssätt, fysikaliska fenomen som finns tillgänglig i barnens litteratur, vad pedagogerna anser vara lämplig litteratur och exempel på hur litteraturen kan göras interaktiv. I diskussionen så diskuteras vad som egentligen kan räknas som litteratur, olika sätt att använda litteraturen i pedagogiskt arbete med fysisk, likheter mellan dessa arbetssätt, lämplig litteratur och genus, samt hur allt knyter an till syftet. / In this study different ways to work with physics through literature are researched. The method was to interview preschool teachers and a representative from the project Lilla bokbryggan, and literature analyzes. The result shows different approaches, Physical phenomena available in children’s literature, what the teachers consider suitable literature and examples on how to make literature interactive. In the discussion I discuss what is actually considered literature, different ways of using the literature in an educational approach in regards to physics, similarities between the different approaches, suitable literature and gender equality, and also how it all ties together with the purpose.

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<p>This essay tries to show how The Swedish Museum of Architecture, Stockholm, with the exibition Architecture in Sweden - Function, Design and Aesthetic through the Ages, presents and represents architecture. It is stated that this is done in a multi-perspectival, multimedial fashion, with the aid of, for example, photography, models and mixed material surrounding the wide concept of ’architecture’. One chapter discusses the relationships between the exhibition on site, the exhibition catalog and the museum´s website. Another chapter argues that there can be no essential or perfect representation of architecture, although this utopian wish most certainly exists even today. A general idea is that the problems regarding architectural representation has something to do with its double position within art history as well as its multi-dimensional characteristics. In addition to this, our daily encounters with architecture is also said to contribute. Furthermore, it is argued that exhibitions understood as site-specific and temporal events, need further research.</p>

This essay tries to show how The Swedish Museum of Architecture, Stockholm, with the exibition Architecture in Sweden - Function, Design and Aesthetic through the Ages, presents and represents architecture. It is stated that this is done in a multi-perspectival, multimedial fashion, with the aid of, for example, photography, models and mixed material surrounding the wide concept of ’architecture’. One chapter discusses the relationships between the exhibition on site, the exhibition catalog and the museum´s website. Another chapter argues that there can be no essential or perfect representation of architecture, although this utopian wish most certainly exists even today. A general idea is that the problems regarding architectural representation has something to do with its double position within art history as well as its multi-dimensional characteristics. In addition to this, our daily encounters with architecture is also said to contribute. Furthermore, it is argued that exhibitions understood as site-specific and temporal events, need further research.

I denna studie undersöker jag barn i förskolan mellan fyra och fem år. Vad de har för tankar i ett möte med för dem ett okänt djur, vattenloppan. Aktiviteten där jag även ställt frågor till barnen angående bl.a. vattenloppans utseende och livsmiljö. Samtidigt som jag filmat har jag kunnat observera två grupper som möter dessa djur antingen med hjälp av bild och video eller med verkliga djur. Genom att barnen fick dokumentera sitt möte med att måla vad de varit med om har jag kunnat få en bild om hur barnen uppfattat djuren. Resultatet visa att flera barn i gruppen som fick uppleva de riktiga djuren kunde detaljera sina upplevelser via deras teckningar. Bildgruppen visade dessutom sämre perceptionsförmåga då de hade en vag bild om vattenloppans storlek och att flera barn såg vattenloppans antenner som vassa och farliga tänder. / In this study, I examine children in preschool between four and five years old. What they have in mind in a meeting with them an unknown animal, water flea. The activity where I asked the children about such as the water fleas appearance and habitat while I was filming, I have been able to do an observation in two groups that could watch these animals either by image and video or as living animals. The children got afterwards documenting his meeting with painting, I was able to get a picture of how the children understood the animals. The results show that several children in the group who got to experience the real animals could detail their experiences through their artwork. The Picture group also showed poorer perception since they had a vague idea about the water flues size and that several children misunderstood the water flues antennas for sharp and dangerous teeth.

The complete sequence of mitochondrial DNA from Diuraphis noxia was obtained and characterized. The mitogenome contains a standard set of 13 protein-coding genes, 19 tRNA genes, 2 ribosomal RNA genes. A+T-rich and ?repets? regions in the same order as those of the other analyzed aphids. Comparison to mtDNAs from other Sternorrhyncha species obtained from GenBank revealed possible markers for studies on population differentiation. Phylogenetic analysis using parsimony and maximum likelihood confirmed the classification of Diuraphis noxia into the Aphididae.