Extracellular vesicles (EVs) gained significant interest within the last decade as a new source of biomarkers for the early detection of diseases and as a promising tool for therapeutic applications. As a result, a need for new methods for EV analysis and quantification has elevated. In this work, I apply Extracellular Vesicles Quantitative Capillary Electrophoresis (EVqCE) to determine (i) the apparent molecular weight of RNA in EVs, (ii) the number of intact EVs in a sample, and (iii) the degree of EV degradation after sonication, vortexing, freeze-thaw cycles and long storage. This separation method is demonstrated on EVs isolated from conditioned media of three different cancer cell lines and human urine samples. Here, I utilize capillary zone electrophoresis with laser-induced fluorescent detection to separate intact EVs from DNA and RNA impurities present in the sample. YOYO-1 dye is used to stain all DNA and RNA in the sample. After lysis of EVs with a detergent, encapsulated DNA and RNA are released. After additional RNase treatment of the EVs sample, RNA is enzymatically cleaved, leaving residual DNA only, in order to calculate the RNA concentration from EVs. Therefore, the initial concentration of intact EV is calculated based on the gain of a nucleic acid peak in capillary electrophoresis and an RNA calibration curve. EVqCE works in a dynamic range of EV concentrations from 10⁸ to 10¹⁰ particles/mL. The quantification process can be completed in less than one hour and requires minimum optimization for CE separation.

Notre recherche porte sur l’évaluation du « satisficing organisationnel » du système de pilotage des entreprises à vocation sociale (EVS). Pour cela, nous mobilisons une méthodologie qualitative centrée sur deux études cas enchâssés, dont une en longitunale. Il s’agit de Sos Maintien à Domicile et l’Eveil, deux entreprises associatives dans le champs institutionnel du médico-social. Nous mettons en évidence trois niveaux de résultats. Tout d’abord, nous évaluons, à travers les cartes cognitives, le système de représentations partagées permettant d’évaluer le sentiment d’efficacité organisationnelle. Ensuite , à travers les comptes de surplus de productivité global, nous mesurons la performance économique et sociale. La convergence des deux premiers niveaux de résultats permet de statuer sur l’état positif ou négatif du satisficing organisationnel. Enfin, l’analyse des cas et leur confrontation avec le modèle conceptuel et la revue de littérature, nous permet de définir des variables essentielles dont le système de pilotage de l’EVS, représenté par le pouvoir managérial ,peut s’en servir dans les différentes modes de régulation du projet. / Our research focuses on the evaluation of the "organizational satisficing" of social purpose business (EVS). For this, we mobilize a qualitative methodology centered on two embedded case studies, including one in longitunale way. "Sos Maintien à Domicile" and "Eveil" are both two associative enterprises in the institutional medico-social care field. We highlight three levels of results. First, we evaluate, through the cognitive maps, the system of shared representations to assess the sense of organizational effectiveness. Then, through the global productivity surplus accounts, we measure the economic and social performance. The convergence of the first two levels of results allows to rule on the positive or negative state of organizational satisficing. Finally, the analysis of cases and confrontation with the conceptual model and the literature review, allows us to identify key variables with the EVS control system, represented by the managerial authority, can be used in various project control modes.

The security dilemma is a well-known model for describing how states act in certain situations due to the lack of trust in the intentions of other states. Instead of trusting that other states will cooperate, and that their increased military spending are for defense purposes, one might conclude that the wisest course of action would be to increase one’s own defenses as well, thus creating a vicious cycle of an arms race. Whether or not one state trusts another seems to be the fundamental aspect of the dilemma. Can a similar pattern be found in the behavior of individuals when it comes to trust? Are their opinions on the need for a stronger defense related to the extent to which they trust others? This is what this thesis strives to answer by examining the correlation between the trust and people’s opinion on defense spending. The thesis is based on previous studies and theories within the field of trust research and attempts to fill the gaps in an otherwise well researched field through analysis of data collected by the research program EVS. The findings of the study indicate a correlation between the general trust of Swedes and their opinion on the importance of maintaining a strong defense.

A degradação lisossomal de proteínas de membrana endocitadas ocorre por meio do direcionamento destas proteínas para vesículas intralumenais (ILVs), formadas no lúmen dos corpos multivesiculares (MVBs), e subsequente fusão dos MVBs com lisossomos. Apesar de sua importância na degradação de proteínas transmembrana, os MVBs possuem outra importante função, a de produzir e liberar vesículas extracelulares (EVs). Neste processo os MVBs não se fundem com lisossomos, mas sim com a membrana plasmática o que resulta na liberação das vesículas residentes no interior dos MVBs para o meio extracelular. Diversas proteínas participam do direcionamento de cargas para os MVBs. Os estudos que delinearam a via de tráfego mediada por AP1 concentraram-se nos complexos contendo a subunidade ?1 que medeia o transporte de proteínas entre a rede trans-Golgi (TGN) e endossomos. Contudo, o genoma humano codifica uma segunda isoforma desta subunidade, denominada ?2, e evidências presentes na literatura e também observadas por nosso grupo indicam que AP1?2 pode regular uma via de tráfego distinta da via classicamente atribuída a AP1. Utilizando ensaios de uptake de EGF em condições onde foi realizado o KD de ?1 ou ?2, foi observado que o silenciamento de ?2 prejudica a degradação de EGF internalizado por seu receptor. Efeito também observado para o próprio receptor de EGF (EGFR) em ensaios de biotinilação da superfície celular. Demonstrando que a degradação lisossomal do complexo EGF-EGFR pela via canônica dos MVBs requer o complexo AP1?2, mas não AP1?1. Em conjunto com este estudo também foi analisado o mecanismo molecular de direcionamento da proteína Nef do HIV-1 para os MVBs associados a liberação de EVs. A proteína Nef do HIV é determinante na modulação do ambiente intracelular favorecendo a replicação do vírus e progressão à AIDS. Nef é ativamente secretado em EVs e sua liberação pode levar a apoptose de células vizinhas aceptoras dessas vesículas. Nef também medeia a redução dos níveis de CD4 e moléculas de MHC-I em EVs. Ainda não é conhecido o mecanismo molecular utilizado por Nef para ser exportado em EVs, mas sabe-se que Nef interage fisicamente com a proteína II acessória da maquinaria ESCRT, Alix, importante no processo de formação das ILVs e seleção das cargas que serão internalizadas nos MVBs. EVs coletadas de células HeLa e linfócitos T CD4+ silenciados para Alix demostraram reduções significativas na liberação de Nef. Estes resultados indicam que Nef requer Alix para sua eficiente liberação em EVs. / Lysosomal degradation of endocytosed membrane proteins occurs through the targeting of these proteins to intraluminal vesicles (ILVs), formed in the multivesicular bodies (MVBs) lumen, and the subsequent fusion of MVBs with lysosomes. Despite its importance in the degradation of transmembrane proteins, MVBs have another important function, the production and release of extracellular vesicles (EVs). In this process, the MVBs do not fuse with lysosomes, but fuse with the plasma membrane resulting in the release of these vesicles that reside within MVBs to the extracellular environment. Several proteins regulate the targeting of cargo to MVBs. Studies that delineated the functions of AP1 in protein trafficking, focused on complexes containing the ?1 subunit, which mediates transport between trans-Golgi network (TGN) and endosomes. However, the human genome encodes a second isoform of this subunit, named ?2. Evidences from the literature, as well as results from our research group, indicate that AP1?2 regulates transport pathways that are distinct from the pathways classically attributed to AP1. By performing EGF-uptake assays under ?1 or ?2 knockdown (KD) conditions, it was observed that ?2 is required for degradation of internalized EGF, effect also observed for the EGF receptor (EGFR) using cell surface biotinylation assays. These results demonstrate that the lysosomal degradation of the EGFEGFR complexes via the canonical MVBs pathway requires the AP1?2 complex, but not AP1?1. In parallel with this study, we also analyzed the molecular mechanism of HIV-1 Nef targeting to MVBs associated with the EVs release. Nef is an important determinant in the modulation of the intracellular environment for efficient HIV replication and progression to AIDS. Nef is actively secreted via EVs and its release may lead to apoptosis of bystander acceptor cells. Moreover, Nef reduces the levels of CD4 and MHC-I molecules in EVs. Despite the importance of Nef release in EVs, the molecular mechanism used by Nef to be exported via EVs is unknown. Nef physically interacts with the ESCRT machinery accessory protein Alix, an important player in the process of ILVs formation and cargo selection. EVs released from HeLa cells and CD4+ T lymphocytes under Alix KD conditions demonstrated a significant IV reduction in Nef release via EVs. These results indicate that Nef requires Alix for its efficient release in EVs.

A degradação lisossomal de proteínas de membrana endocitadas ocorre por meio do direcionamento destas proteínas para vesículas intralumenais (ILVs), formadas no lúmen dos corpos multivesiculares (MVBs), e subsequente fusão dos MVBs com lisossomos. Apesar de sua importância na degradação de proteínas transmembrana, os MVBs possuem outra importante função, a de produzir e liberar vesículas extracelulares (EVs). Neste processo os MVBs não se fundem com lisossomos, mas sim com a membrana plasmática o que resulta na liberação das vesículas residentes no interior dos MVBs para o meio extracelular. Diversas proteínas participam do direcionamento de cargas para os MVBs. Os estudos que delinearam a via de tráfego mediada por AP1 concentraram-se nos complexos contendo a subunidade ?1 que medeia o transporte de proteínas entre a rede trans-Golgi (TGN) e endossomos. Contudo, o genoma humano codifica uma segunda isoforma desta subunidade, denominada ?2, e evidências presentes na literatura e também observadas por nosso grupo indicam que AP1?2 pode regular uma via de tráfego distinta da via classicamente atribuída a AP1. Utilizando ensaios de uptake de EGF em condições onde foi realizado o KD de ?1 ou ?2, foi observado que o silenciamento de ?2 prejudica a degradação de EGF internalizado por seu receptor. Efeito também observado para o próprio receptor de EGF (EGFR) em ensaios de biotinilação da superfície celular. Demonstrando que a degradação lisossomal do complexo EGF-EGFR pela via canônica dos MVBs requer o complexo AP1?2, mas não AP1?1. Em conjunto com este estudo também foi analisado o mecanismo molecular de direcionamento da proteína Nef do HIV-1 para os MVBs associados a liberação de EVs. A proteína Nef do HIV é determinante na modulação do ambiente intracelular favorecendo a replicação do vírus e progressão à AIDS. Nef é ativamente secretado em EVs e sua liberação pode levar a apoptose de células vizinhas aceptoras dessas vesículas. Nef também medeia a redução dos níveis de CD4 e moléculas de MHC-I em EVs. Ainda não é conhecido o mecanismo molecular utilizado por Nef para ser exportado em EVs, mas sabe-se que Nef interage fisicamente com a proteína II acessória da maquinaria ESCRT, Alix, importante no processo de formação das ILVs e seleção das cargas que serão internalizadas nos MVBs. EVs coletadas de células HeLa e linfócitos T CD4+ silenciados para Alix demostraram reduções significativas na liberação de Nef. Estes resultados indicam que Nef requer Alix para sua eficiente liberação em EVs. / Lysosomal degradation of endocytosed membrane proteins occurs through the targeting of these proteins to intraluminal vesicles (ILVs), formed in the multivesicular bodies (MVBs) lumen, and the subsequent fusion of MVBs with lysosomes. Despite its importance in the degradation of transmembrane proteins, MVBs have another important function, the production and release of extracellular vesicles (EVs). In this process, the MVBs do not fuse with lysosomes, but fuse with the plasma membrane resulting in the release of these vesicles that reside within MVBs to the extracellular environment. Several proteins regulate the targeting of cargo to MVBs. Studies that delineated the functions of AP1 in protein trafficking, focused on complexes containing the ?1 subunit, which mediates transport between trans-Golgi network (TGN) and endosomes. However, the human genome encodes a second isoform of this subunit, named ?2. Evidences from the literature, as well as results from our research group, indicate that AP1?2 regulates transport pathways that are distinct from the pathways classically attributed to AP1. By performing EGF-uptake assays under ?1 or ?2 knockdown (KD) conditions, it was observed that ?2 is required for degradation of internalized EGF, effect also observed for the EGF receptor (EGFR) using cell surface biotinylation assays. These results demonstrate that the lysosomal degradation of the EGFEGFR complexes via the canonical MVBs pathway requires the AP1?2 complex, but not AP1?1. In parallel with this study, we also analyzed the molecular mechanism of HIV-1 Nef targeting to MVBs associated with the EVs release. Nef is an important determinant in the modulation of the intracellular environment for efficient HIV replication and progression to AIDS. Nef is actively secreted via EVs and its release may lead to apoptosis of bystander acceptor cells. Moreover, Nef reduces the levels of CD4 and MHC-I molecules in EVs. Despite the importance of Nef release in EVs, the molecular mechanism used by Nef to be exported via EVs is unknown. Nef physically interacts with the ESCRT machinery accessory protein Alix, an important player in the process of ILVs formation and cargo selection. EVs released from HeLa cells and CD4+ T lymphocytes under Alix KD conditions demonstrated a significant IV reduction in Nef release via EVs. These results indicate that Nef requires Alix for its efficient release in EVs.

<p>Det här är en studie som belyser programmet Europeisk korttidsvolontärtjänst (St EVS) samt granskar om programmet kan vara lämpligt vid vägledning av unga i behov av särskilt stöd.</p><p>Litteraturstudier och intervjuer med tre personer som har erfarenhet av arbetssättet har genomförts. Forskningsfrågorna ger svar på vilka argument som finns för att använda programmet, hur arbetet med St EVS går till samt vilka ungdomar som kan delta.</p><p>Tidigare forskning inom området är näst in till obefintlig, vidare saknas kännedom om St EVS bland många yrkesgrupper som arbetar med unga i behov av särskilt stöd.</p><p>I resultatet framkommer att St EVS ökar ungdomarnas självförtroende och främjar deras personliga utveckling. Resultatet tyder också på att det ger motivation för arbete eller studier.</p><p>Andra fördelar är att programmet är gratis för ungdomarna och erbjuder mycket trygghet.</p><p>Resultaten pekar dock mot att det är viktigt med förberedelser och att sändande organisationen känner till och har förtroende för mottagande organisationen. Förutsatt att mottagande organisationen kan ge ungdomen det individuella stöd som han/hon behöver, kan ungdomar med olika förutsättningar och behov delta i programmet.</p><p>Analys av resultaten indikerar att programmet skulle kunna innebära en ekonomisk vinst både på kort och på lång sikt för kommuner. En annan slutsats är att fler samarbeten borde finnas där olika yrkesgrupper arbetar tillsammans med St EVS.</p><p>Vidare kan programmet anses lämpligt att använda i vägledning av ungdomar i behov av särskilt stöd. Då ensamt arbete med programmet tar mer tid och engagemang är samarbete att föredra. Förutsatt att ett samarbete finns är det troligt att studie- och yrkesvägledare både kan tillföra mycket och själva tjäna på ett sådant samarbete.</p><p>Resultatet av studien kan vara till nytta för studie- och yrkesvägledare på gymnasium, särgymnasium och arbetsförmedlingar, samt för kommuner och för övriga yrkesgrupper som arbetar med ungdomar i behov av särskilt stöd.</p>

Det här är en studie som belyser programmet Europeisk korttidsvolontärtjänst (St EVS) samt granskar om programmet kan vara lämpligt vid vägledning av unga i behov av särskilt stöd. Litteraturstudier och intervjuer med tre personer som har erfarenhet av arbetssättet har genomförts. Forskningsfrågorna ger svar på vilka argument som finns för att använda programmet, hur arbetet med St EVS går till samt vilka ungdomar som kan delta. Tidigare forskning inom området är näst in till obefintlig, vidare saknas kännedom om St EVS bland många yrkesgrupper som arbetar med unga i behov av särskilt stöd. I resultatet framkommer att St EVS ökar ungdomarnas självförtroende och främjar deras personliga utveckling. Resultatet tyder också på att det ger motivation för arbete eller studier. Andra fördelar är att programmet är gratis för ungdomarna och erbjuder mycket trygghet. Resultaten pekar dock mot att det är viktigt med förberedelser och att sändande organisationen känner till och har förtroende för mottagande organisationen. Förutsatt att mottagande organisationen kan ge ungdomen det individuella stöd som han/hon behöver, kan ungdomar med olika förutsättningar och behov delta i programmet. Analys av resultaten indikerar att programmet skulle kunna innebära en ekonomisk vinst både på kort och på lång sikt för kommuner. En annan slutsats är att fler samarbeten borde finnas där olika yrkesgrupper arbetar tillsammans med St EVS. Vidare kan programmet anses lämpligt att använda i vägledning av ungdomar i behov av särskilt stöd. Då ensamt arbete med programmet tar mer tid och engagemang är samarbete att föredra. Förutsatt att ett samarbete finns är det troligt att studie- och yrkesvägledare både kan tillföra mycket och själva tjäna på ett sådant samarbete. Resultatet av studien kan vara till nytta för studie- och yrkesvägledare på gymnasium, särgymnasium och arbetsförmedlingar, samt för kommuner och för övriga yrkesgrupper som arbetar med ungdomar i behov av särskilt stöd.

This diploma thesis is focused to systems used for automatic control of civil aircraft along an precision approach trajectory. It describes precision approach systems used in the past and their evolutional steps leading to their current and future configuration. The thesis does not put the goal to be a detailed study but to be a publication providing to pilots and students of Aeronautical traffic the introduction to the field of precision approach systems in context to another fields of aeronautical traffic, describing its current status and infer its further development.

With the ever-pressing threat of a climate crisis, the EU has decided to become the first climate-neutral continent by 2050. This in turn will require the road transportation sector to make a transition from fossil dependent to fossil-free vehicles. Sweden has the objective to become net positive in GHG emissions by 2045. To be on track to reach this goal, the GHG emissions of the domestic transport sector must be reduced by 70% by 2030 compared to 2017’s levels. Electric vehicles (EVs) are leading the way in the transition to fossil-free vehicles. To further springboard the diffusion of EVs, the development of a fully functional EV charging network is required. In order to assist the transition to electric vehicles, this report aims to analyse the development of the public fast charging infrastructure in Norrland and Svealand from now to 2030. Additionally, identify geographical areas where an expansion of the public EV fast charging network is needed to cover the future demand of electrified passenger cars. However, there are two major hurdles in building a fast charging network with full coverage. The first is the high monthly costs of providing fast charging which needs a certain utilisation rate to cover the expenses. The second hurdle is the difficulty to receive a grid connection, in certain areas, at the required power output to be able to provide EV fast charging. Therefore, a semi-mobile battery solution used for EV charging is analysed through a business model perspective. The semi-mobile battery solution requires a lower grid connection hence it could be possible to implement public EV fast charging at a lower monthly cost and to develop the public EV fast charging network in otherwise technical difficult areas. A mixed-method approach including both quantitative and qualitative elements was utilised. Primarily, a study of 10 interviews with respondents from a range of different fields connected to EV charging and batteries was performed in combination with a literature review and document analysis. In addition, existing traffic flow data and data of fast-charging infrastructure, were converged via ArcGIS Pro to illustrate the coverage of the fast charging network. Furthermore, projections of the development of the EV fleet were used in order to forecast the flow of EVs in Norrland and Svealand by 2030. Based on these forecasts the future demand of public EV fast charging was analysed. Resulting in a map showing areas of interest, where there will arise a need to expand the charging infrastructure. These areas are Umeå to Piteå, Lycksele with proximity, Bollnäs to Ljusdal and Leksand to Älvdalen. Additionally, the exiting public fast charging infrastructure was identified to require expansion of existing charging stations due to the increased traffic flow of EVs by 2030. The upgrade of existing stations was further assessed to be required to meet both a permanent and seasonal demand, hence making semi-mobile battery supported charging an attractive solution. Furthermore, the design of a semi-mobile battery supporting public EV fast charging was identified to be influenced by situational aspects and that the location-specific conditions were vital in determining profitability for a specific case. For example, the power output in the EV chargers should be adapted to the specifications of the geographical location and the customer segment identified. The energy storage capacity of the battery should also be designed based on the conditions of the location. A connection to the electricity grid exceeding 0.1 MW was also important since it enables the semi-mobile battery to provide additional services to the electricity grid and hence increase revenue streams. Furthermore, FCR-D Up was determined to be the most suitable complementary service to integrate into the system. One major challenge for the semi-mobile battery, based on a business model perspective, is the high costs for semi-mobile batteries and EV fast charging station hardware. However, these costs are projected to continue to decrease and consequently, improve the opportunities for semi-mobile lithium-ion batteries.

No description available.