<p>Mast cells are long-lived effector cells of importance for both acute and chronic inflammations. Mast cells can be activated in many different ways, leading to the release of inflammatory mediators. In contrast to most other inflammatory cells, activated mast cells have the capacity to recover, regranulate and thereby be activated again. </p><p>In this thesis I have investigated the mechanisms involved in regulating activation-induced mast cell survival. We have found that cross-linking of FcεRI-bound IgE with an antigen (IgER-CL) induces a survival program in mast cells. Upon IgER-CL, mouse and human mast cells upregulate the pro-survival Bcl-2 family gene A1/Bfl-1. A1^-/- mast cells degranulate upon FcεRI activation but they cannot recover most likely due to the lack of A1. Sensitized and provoked A1^-/- mice exhibit lower amounts of mast cells compared to littermate controls. In contrast to mast cells, no Bfl-1 expression or survival promotion can be detected in basophils after IgER-CL. Another mast cell secretagogue, an adenosine receptor agonist, neither promoted upregulation of A1 nor survival.</p><p>Although it is well established that mast cell survival is dependent on stem cell factor (SCF), it has not been described how this process is regulated. We have found that SCF promotes survival through Akt-mediated inhibition of the forkhead transcription factor FOXO3a and its transcriptional target Bim, a BH3-only pro-apoptotic protein. SCF-treatment prevents upregulation of Bim protein expression and leads to an upregulation of Bim phosphorylation through PI3-kinase and MEK-dependent pathways. Overexpression of FOXO3a causes an upregulation of Bim and induces mast cell apoptosis, even in the presence of SCF. </p><p>Taken together, the work in this thesis demonstrates that A1/Bfl-1 and Bim play key roles in mast cell survival. These findings might be of importance in understanding the mechanisms of mast cell longevity and hence for possible new therapeutics used for mast cell-associated inflammations.</p>

Mast cells are long-lived effector cells of importance for both acute and chronic inflammations. Mast cells can be activated in many different ways, leading to the release of inflammatory mediators. In contrast to most other inflammatory cells, activated mast cells have the capacity to recover, regranulate and thereby be activated again. In this thesis I have investigated the mechanisms involved in regulating activation-induced mast cell survival. We have found that cross-linking of FcεRI-bound IgE with an antigen (IgER-CL) induces a survival program in mast cells. Upon IgER-CL, mouse and human mast cells upregulate the pro-survival Bcl-2 family gene A1/Bfl-1. A1-/- mast cells degranulate upon FcεRI activation but they cannot recover most likely due to the lack of A1. Sensitized and provoked A1-/- mice exhibit lower amounts of mast cells compared to littermate controls. In contrast to mast cells, no Bfl-1 expression or survival promotion can be detected in basophils after IgER-CL. Another mast cell secretagogue, an adenosine receptor agonist, neither promoted upregulation of A1 nor survival. Although it is well established that mast cell survival is dependent on stem cell factor (SCF), it has not been described how this process is regulated. We have found that SCF promotes survival through Akt-mediated inhibition of the forkhead transcription factor FOXO3a and its transcriptional target Bim, a BH3-only pro-apoptotic protein. SCF-treatment prevents upregulation of Bim protein expression and leads to an upregulation of Bim phosphorylation through PI3-kinase and MEK-dependent pathways. Overexpression of FOXO3a causes an upregulation of Bim and induces mast cell apoptosis, even in the presence of SCF. Taken together, the work in this thesis demonstrates that A1/Bfl-1 and Bim play key roles in mast cell survival. These findings might be of importance in understanding the mechanisms of mast cell longevity and hence for possible new therapeutics used for mast cell-associated inflammations.

<p>Validerat; 20130215 (global_studentproject_submitter)</p>

Orientador: Lucilene Delazari Santos / Resumo: O veneno de serpente tem sido utilizado para diversas finalidades terapêuticas; portanto, desde o início do século XX, a criação de serpentes em cativeiro tornou-se uma atividade cada vez mais relevante. Os principais motivos da existência e permanência destes cativeiros se atribuem à produção dos soros antiofídicos, e o uso de seus venenos e seus componentes isolados com potenciais aplicações à saúde animal e humana. Porém, a vida em cativeiro pode resultar na síndrome da má adaptação, a qual o animal sob estresse desenvolve inapetência e, consequentemente há o acometimento da imunidade, podendo desenvolver infecções por micro-organismos como fungos, bactérias e vírus. Desta forma, o controle e prevenção de infecções em serpentes mantidas em cativeiro tornaram-se assuntos muito importantes. Dentre as doenças infecciosas virais, têm-se a infecção pelo Paramixovírus ofídico, atualmente, denominado Ferlavirus reptiliano. Este vírus é altamente devastador em serpentes, pois atua no sistema nervoso central e respiratório, podendo levar à morte e/ou morbidade na fase aguda da doença, sendo que na fase crônica, o animal pode atuar como reservatório do vírus, e disseminar a doença no plantel. Atualmente, não há tratamento específico nem vacinas disponíveis para esta virose. Porém, testes sorológicos estão emergindo para a detecção de anticorpos. Dentre estes testes, encontram-se o ensaio de Inibição de Hemaglutinação (HI) e o ensaio de ELISA de Bloqueio de Fase Líquida (BFL-ELISA), ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Snake venom has been used for several therapeutic purposes. Therefore, breeding snakes in captivity has become an increasingly relevant activity since early20th century. The main reasons for the existence and permanence of these captivities are the production of snake antivenom and the use of venom and its isolate compounds with potential applications in human and animal health. However, living in captivity may result in the maladaptation syndrome, in which the animal under stress develops inappetence and, consequently, a decrease in immunity, becoming more likely to develop infections caused by microorganisms as fungi, bacteria and viruses. Thus, the control and prevention of infections in snakes kept in captivity became very important subjects. The infection by the ophidian paramyxovirus named reptilian ferlavirus is among the viral infectious diseases. This virus is highly devastating in snakes because its action in central and respiratory nervous system and may lead to death and/or morbidity in the acute stage of the disease, consideringthat the animal may act as a reservoir of the virus in the chronic stage and spread the disease in the breeding stock. Nowadays, there isn’t a specific treatment or vaccines available for strike this viral infection, however, serological tests are emerging for antibodies detection. Hemagglutination Inhibition (HI) and Liquid Phase Blocking ELISA (LPB-ELISA) are among these tests, which differs from each other in sensitivity and analytical ... (Complete abstract click electronic access below) / Mestre

Le virus humain T lymphotrope de type 1 (HTLV-1) est l'agent étiologique de la leucémie/lymphome T de l'adulte (ATLL) qui se développe après plusieurs décennies et pour laquelle il n'existe à ce jour pas de traitement efficace. Parmi les protéines virales de HTLV-1, Tax et HBZ jouent un rôle déterminant dans le développement de l'ATLL. Si Tax participe au processus leucémogène dès les étapes précoces, HBZ jouerait plutôt un rôle dans le maintien du phénotype tumoral dans les étapes tardives. Dans ce contexte, là nous nous sommes intéressés à la régulation de l'expression des protéines anti-apoptotiques Bfl-1 et Bcl-xL, par les protéines virales Tax et HBZ. Nous avons montré que Tax induit l'expression des protéines anti-apoptotiques Bfl-1 et Bcl-xL de la famille Bcl-2 via la voie NF-B, alors que HBZ n'a aucun effet sur leur expression. De plus, Tax coopère avec les facteurs de transcription c-Jun et JunD de la voie AP-1 pour augmenter l'expression de ces gènes anti-apoptotiques. En revanche, HBZ module uniquement la trans-activation de bfl-1 induite par Tax. L'ensemble de nos résultats indique donc que Tax joue un rôle prépondérant dans l'activation de l'expression de Bfl-1 et de Bcl-xL et suggère que Bfl-1 et Bcl-xL sont exprimées au cours des étapes précoces et tardives du développement de l'ATLL. Par une stratégie d'ARN interférence, nous avons ensuite montré que Bfl-1 et/ou Bcl-xL sont impliquées dans la survie de lignées cellulaires T infectées par HTLV-1, suggérant que Bfl-1 et Bcl-xL représentent des cibles thérapeutiques potentielles pour traiter l'ATLL. Actuellement, il existe des petites molécules ciblant les membres anti-apoptotiques de la famille Bcl-2, mais aucune ne cible spécifiquement Bfl-1. En collaboration avec la société IMAXIO, nous avons identifié par deux cribles à haut débit 83 molécules capables d'inhiber l'activité anti-apoptotique de Bfl-1. L'une de ces molécules induit spécifiquement la mort de lignées cellulaires T infectées par HTLV-1 pour lesquelles Bfl-1 représente un gène de survie. Ainsi, ce travail doit permettre à terme de développer de futurs médicaments dirigés contre Bfl-1 et de proposer une nouvelle stratégie thérapeutique ciblée contre l'ATLL

Le virus humain T lymphotrope de type 1 (HTLV-1) est l’agent étiologique de la leucémie/lymphome T de l’adulte (ATLL) qui se développe après plusieurs décennies et pour laquelle il n’existe à ce jour pas de traitement efficace. Parmi les protéines virales de HTLV-1, Tax et HBZ jouent un rôle déterminant dans le développement de l’ATLL. Si Tax participe au processus leucémogène dès les étapes précoces, HBZ jouerait plutôt un rôle dans le maintien du phénotype tumoral dans les étapes tardives. Dans ce contexte, là nous nous sommes intéressés à la régulation de l’expression des protéines anti-apoptotiques Bfl-1 et Bcl-xL, par les protéines virales Tax et HBZ. Nous avons montré que Tax induit l’expression des protéines anti-apoptotiques Bfl-1 et Bcl-xL de la famille Bcl-2 via la voie NF-κB, alors que HBZ n’a aucun effet sur leur expression. De plus, Tax coopère avec les facteurs de transcription c-Jun et JunD de la voie AP-1 pour augmenter l’expression de ces gènes anti-apoptotiques. En revanche, HBZ module uniquement la trans-activation de bfl-1 induite par Tax. L’ensemble de nos résultats indique donc que Tax joue un rôle prépondérant dans l’activation de l’expression de Bfl-1 et de Bcl-xL et suggère que Bfl-1 et Bcl-xL sont exprimées au cours des étapes précoces et tardives du développement de l’ATLL. Par une stratégie d’ARN interférence, nous avons ensuite montré que Bfl-1 et/ou Bcl-xL sont impliquées dans la survie de lignées cellulaires T infectées par HTLV-1, suggérant que Bfl-1 et Bcl-xL représentent des cibles thérapeutiques potentielles pour traiter l’ATLL. Actuellement, il existe des petites molécules ciblant les membres anti-apoptotiques de la famille Bcl-2, mais aucune ne cible spécifiquement Bfl-1. En collaboration avec la société IMAXIO, nous avons identifié par deux cribles à haut débit 83 molécules capables d’inhiber l’activité anti-apoptotique de Bfl-1. L’une de ces molécules induit spécifiquement la mort de lignées cellulaires T infectées par HTLV-1 pour lesquelles Bfl-1 représente un gène de survie. Ainsi, ce travail doit permettre à terme de développer de futurs médicaments dirigés contre Bfl-1 et de proposer une nouvelle stratégie thérapeutique ciblée contre l’ATLL / Human T lymphotropic virus type 1 (HTLV-1) is the etiological agent of adult T-cell leukemia/lymphoma (ATLL) that develops after several decades and for which there is no effective treatment. Among the viral proteins of HTLV-1, Tax and HBZ play a major role in the development of ATLL. If Tax participates in the initiation of leukemogenesis from the early stages, HBZ rather plays a role in maintaining the tumor phenotype in the late stages. The aims of our study were to better understand the regulation of Bfl-1 and Bcl-xL anti-apoptotic protein expression by Tax and HBZ viral proteins, as well as their role in the survival of HTLV-1-infected T-cells to propose new therapeutic strategies. We showed that Tax induces Bfl-1 and Bcl-xL expression via the NF-κB pathway, whereas HBZ has no effect on their expression. Tax also cooperates with c-Jun and JunD transcription factors of AP-1 family to increase the expression of these anti-apoptotic genes. By contrast, HBZ modulates the Tax-induced bfl-1 trans-activation. Altogether, our data indicate that Tax plays a key role in activating Bfl-1 and Bcl-xL expression and suggests that Bfl-1 and Bcl-xL are potentially expressed during the early and the late stages of ATLL development. Using short hairpin RNA strategy, we then showed that Bfl-1 and/or Bcl-xL are involved in HTLV-1-infected T-cell line survival, indicating that Bfl-1 and Bcl-xL represent potential therapeutic targets in the case of ATLL. One approach currently being developed in anti-cancer drug discovery is to search for small inhibitory compounds targeting anti-apoptotic proteins of the Bcl-2 family. But so far, no drug specifically targeting Bfl-1 is available. In collaboration with the IMAXIO Company, we have identified 83 molecules able to inhibit Bfl-1 anti-apoptotic activity using two high-throughput screening. One of these molecules specifically induced the death of HTLV-1-infected T-cell for which Bfl-1 represents a survival gene. This work provides new insight for long-term development of future drugs directed against Bfl-1 and should allow us to propose new therapeutic strategy for ATLL treatment

Inom EU pågår ett ständigt arbete med harmonisering av lagar i syfte att utforma och införa lagstiftning av högsta kvalitet, samtidigt som hänsyn tas till att de administrativa bördorna står i proportion till deras nytta. Med detta som bakgrund har EU arbetat med att främja lättnader för små och medelstora företag på internationell såväl som nationell nivå, vilket resulterade i en revision av de egna direktiven. Som en följd av Europaparlamentet och Rådets direktiv av 2013 gällande årsbokslut, koncernredovisning och rapporter i vissa typer av företag, tillsattes i slutet av 2012 en utredning av regeringen. Utredningens uppgift var att lämna förslag till de lagbestämmelser som i svensk rätt skulle krävas för att genomföra direktivet. Utöver själva direktivet behandlade utredningen även frågan om de befintliga årsredovisningslagarna bör utformas på ett mer överskådligt och tydligt sätt. Utredningen resulterade i ett delbetänkande som behandlade genomförandet av EU:s nya redovisningsdirektiv, samt ett slutbetänkande som innefattade en översyn av årsredovisningslagarna. Remiss på slutbetänkandet skickades ut till 57 instanser och svaren på denna skulle vara Justitiedepartementet tillhanda senast den 1 december 2015. Totalt inkom det svar från 36 instanser gällande slutbetänkandet.Syftet med studien är att undersöka och redogöra för de attityder de olika remissinstanserna har till de förslag som angivits av utredningen. Genom att skapa en modifierad intressentmodell kategoriserar vi instanserna i olika intressentgrupper och undersöker om det finns samband i remissvaren baserat på vilken intressentgrupp respektive instans är kategoriserad i, samt vilka frågor som är av störst vikt för instanserna. Studien är av kvalitativ karaktär och empirin är baserad på de inkomna remissvaren.Studien visar att de flesta remissinstanserna ser positivt på ett införande av en ny företagskategori, mikroföretag. Då den svenska utredningen föreslår lägre gränsvärden för denna kategori än vad EU-direktivet gör, gav detta upphov till diskussion. Utredningen menar att det underlättar att ha samma gräns som för revisionsplikt, medan de som förespråkar högre gränsvärden anser att det medför mer svårigheter för svenska företag jämfört med deras europeiska jämlikar. Även den kostnadsbesparing som presenterats av utredningen ifrågasätts av ett flertal instanser då de anser att kalkylen är för snävt beräknad, och menar att kostnaderna kommer att bli högre både för det enskilda företaget och för samhället. Gällande moderniseringen av ÅRL och BFL är alla instanser överens om att det är något som ligger i tiden, baserat på att lagarna idag är allt för komplexa och saknar logisk struktur. Förslagen gällande fastställandet av årsredovisningen samt fastställelseintyget har renderat i minst yttranden från remissinstanserna. De instanser som svarat är också de som påverkas mest av de nya reglerna.De slutsatser vi drar är att de flesta instanserna är positiva till de förslag som presenterats av utredningen. Debatten angående gränsdragningen kommer med stor sannolikhet att fortgå även efter det att kategorin mikroföretag har införts i lagstiftningen. Vi anser att det är av vikt att Sverige tillämpar de gränsvärden som EU-direktivet förespråkar, bland annat av harmoniseringsskäl. I kostnadsfrågan ser vi att det i det långa loppet är viktigare att lättnader införs för de mindre företagen än att det på kort sikt kommer innebära merkostnader för myndigheterna. Gällande förslaget till en modernisering av ÅRL och BFL så är vi precis som remissinstanserna positiva till det då vi anser det vara av största vikt att förbättra tydligheten i lagtexten. / Within the EU, the work with harmonization of laws is an on-going process. The aim is to design and implement legislation of the highest quality, while taking into account that the administrative burdens are proportionate to their advantage. Against this background, the EU has worked to promote relief for small and medium-sized entities at the international as well as the national level, resulting in an audit of its own directives. As a result of the European Parliament and Council Directive of 2013 concerning the annual accounts, consolidated financial statements and related reports of certain types, an investigation was added by the government at the end of 2012. The investigators task was to submit proposals to the legislative provisions of Swedish law that would be required to implement the Directive. In addition to the Directive itself, the investigation treated the question whether the existing annual accounts legislation should be redesigned in a more transparent and clear manner. The investigation resulted in an interim report that dealt with the implementation of the new EU accounting directives, as well as a final report, that handled a review of the annual accounts act. The consultation on the final report was sent to 57 agencies and the answer to this was to be at the Ministry of Justice no later than 1 December 2015. Answers were received from 36 agencies.The purpose of this study is to examine and report on the attitudes of the various consultative agencies regarding the proposals set forth by the investigation. By creating a modified stakeholder model, we categorize instances of the various stakeholders and examine whether there is a correlation in the responses based on the stakeholder group each instance is categorized into, and what issues are of most importance to the instances. The study is qualitative in nature and the empirical data is based on the submitted consultation responses.The study shows that most agencies welcome the introduction of a new business category, micro-entities. There are some concerns regarding the limits for this category, since the limits suggested by the Swedish investigation are lower than the limits suggested by the EU. The investigation believes that it is helpful to have the same limit as for auditing duty, while those who advocate higher levels believes it will cause more problems for Swedish companies compared to their European peers. The cost saving estimated by the investigation are believed to be too narrowly calculated by several agencies, who argue that the costs will be higher for both the individual entity as well as for society. Regarding the modernization of the annual accounts acts all agencies agree that it is something that needs to be done, based on that the laws are too complex and lack logical structure. The proposals regarding the establishment of the annual report and approval certificate has rendered the least opinions of the consulted agencies. The replying agencies are the ones most affected by these new rules.The conclusions that we draw are that most of the agencies are positive towards the proposals presented by the investigation. The debate over the limits for micro-entities will likely continue after the category has been introduced to legislation. We believe that it is important for Sweden to apply the values that the EU directive advocates, for harmonization reason among other things. Regarding the cost issue we see that it is more important to promote relief for the smaller companies, than the meaning of increased costs in the short term. Regarding the modernization of the annual account acts we are, just as the consultative agencies, in favour of it because we believe it to be of utmost importance to improve the clarity of the legal text.(This essay is written in Swedish)

I studien undersöks användningen av digital portfolio i bildämnet och hur den digitala portfolion ingår i att synliggöra lärandeprocessen, samt hur den kan skapa en pågående dialog mellan elev och lärare. Vidare undersöks den digitala portfolions roll i elevers delaktighet i bedömningsprocessen. Metoden är kvalitativ och stödjer sig på halvstrukturerade intervjuer med pedogog och elever från högstadiet som använder sig av digital portfolio i bildämnet. Resultatet visar att både pedagog och elever ser möjligheterna med den digitala portfolion vad det gäller synliggörande av elevernas lärprocess i bild. Ett multimodalt arbetssätt är en framgångsfaktor som bidrar både till synligheteten av lärprocess och bedömningsprocess, samt främjar alternativa kommunikationsformer. Hinder i att uppnå en synlighet och delaktighet i lärandeprocessen och bedömningsprocessen kan dels bero på att den digitala portfolion kan betraktas av eleverna som för abstrakt och därmed också negligeras, eller att eleverna inte ser eller har förstått värdet i att dokumentera. Ytterligare en faktor som är nödvändig för att eleverna ska känna sig delaktiga i, bland annat bedömningsprocessen, är att pedagogens feedback måste vara kontinuerlig och effektiv. Ur ett socialkulturellt perspektiv, med fokus på design för lärande, visar sig den digitala portfolion ha stor potential för synlighet och delaktighet i både lär- och bedömningsprocessen. / The study examines the usage of digital portfolio in the art subject and how the digital portfolio is a part of making the learning process visible. It also investigates how it facilitates an ongoing dialogue between student and teacher. In addition, the study aims to investigate the digital portfolio’s roll in making students involved in the assessment proceqss. The method used in the study is qualitative and relies on half structured interviews with a teacher and students from a Swedish högstadie (8th and 9th grade) who use digital portfolio in the art subject. The result indicate that both the teacher and students alike see the possibilities with the digital portfolio concerning visibility of the students’ learning process in the art subject. A factor for success is a multimodal working practice which contributes both to the visibility of the learning process and assessment process, while also encouraging alternative communication channels. Obstacles in accomplishing visibility in the learning and assessment process can be attributed to the students’ perceiving the digital portfolio as too abstract, consequently neglecting it, or that the students do not see the value in documenting. Yet another factor necessary for students to feel involved, especially in the assessment process, is that the teacher’s feedback needs to be continuous and effective. From a sociocultural perspective, with a focus on design for learning, the digital portfolio has proved to have great potential for visibility and participation in both the learning and assessment process.