As técnicas baseadas em microfeixe de íons energéticos possuem aplicações em diferentes áreas de conhecimento, tais como materiais, biologia, neurociência, ciências ambientais, entre outros. A microssonda iônica também possui a capacidade de fabricar micro-estruturas com aplicações em biomateriais, micro-fotônica, micro-fluídica, membranas porosas e diversas outras. O Laboratório de Implantação Iônica do Instituto de Física (LII) da UFRGS (Universidade Federal do Rio Grande do Sul) conta com uma linha de microssonda iônica que dispõe de detectores para a realização de diversos tipos de experimentos: Emissão de RaiosX Induzida por Partículas (Particle Induced X-Ray Emission – PIXE), Espectroscopia de Espalhamento de Rutherford (Rutherford Backskattering Spectroscopy – RBS), Reação Nuclear (Nuclear Reaction Analysis – NRA) e Microscopia Iônica de Varredura por Transmissão (Scanning Transmission Ion Microscopy – STIM). Além disso, tem-se aperfeiçoado a técnica de micro-fabricação denominada Litografia por Feixe de Prótons (Proton Beam Writing – PBW), atualmente em substratos poliméricos. Este trabalho apresenta dois focos: um deles é a análise de fígado de peixe pela técnica de micro-PIXE. Estes fígados apresentam lesões associadas à exposição de contaminantes. Essas lesões são acompanhadas de células pigmentosas à base de metais chamadas de centros de melanomacrófagos. Observou-se que o fígado de peixe é uma amostra em potencial para estudos ambientais. Outro foco foi o estudo da técnica de STIM em películas poliméricas micro-fabricadas por PBW. Foram estudadas diferentes configurações geométricas para a técnica de STIM, bem como a utilização de feixes de H+1, He+2 e Li+3. Diferentes amostras foram preparadas, desde estruturas simples (possuindo apenas orifícios) até amostras multi-estruturadas do tipo degrau. As medidas de STIM foram comparadas com medidas de Microscopia Eletrônica de Varredura (MEV) de maneira a confirmar a viabilidade e confiabilidade dos estudos morfológicos das amostras em questão. A técnica de STIM se apresentou viável para o estudo morfológico de microestruturas fabricadas por microfeixe de íons em películas poliméricas, sendo que a melhor configuração é na geometria on-axis utilizando o íon He+2 que provê melhor resolução em energia, sem comprometer a resolução espacial. Em comparação com medidas de MEV, apresenta vantagens como maior rapidez, preservação da amostra e o poder de resolver estruturas enterradas na amostra. A similaridade entre as duas técnicas com relação às demais características faz do STIM uma excelente alternativa à mais comumente utilizada para diversas análises morfológicas de películas poliméricas. / Ion beam techniques have many applications in different areas such as materials, biology, neuroscience and environmental science among others. The ion microprobe has also the ability of fabricating microstructures with applications in the fields of biomaterials, micro-photonics, micro-fluidics, porous membranes and many others. The Ion Implantation Laboratory of the Physics Institute at UFRGS has a microprobe beamline equiped with detectors used in different techniques: Particle Induced X-Ray Emission (PIXE), Rutherford Backscattering Spectroscopy (RBS), Nuclear Reaction Analysis (NRA) and Scanning Transmission Ion Microscopy (STIM). Besides, its scanning capability makes allows the use of the micro-fabrication technique called Proton Beam Writing (PBW). Currently, this technique has been used to write micro-structures over polymers. Several projects are in progress for the development and improvement of such techniques, since the microbeam line is a recent acquisition by the laboratory. This work presents two guidelines: one of them is the analysis of fish liver through the micro-PIXE technique. These livers present injuries associated to the exposure of contaminants. The injuries are characterized by the presence of pigmented cells based on metals called melanomacrophage centers. It was observed that the fish liver is a promising sample for environmental studies. The other guideline is the study of the STIM technique on micro-fabricated polymer foils by PBW. Different geometric configurations for the STIM technique were explored, as well as the use of different ion beams (H+1, He+2 and Li+3). Different samples were prepared, ranging from simple structures (containing only one hole) to step-like multi-structured samples. STIM measurements were compared with Scanning Electron Microscopy (SEM) in order to confirm the viability and reliability of morphological studies of the samples under study. The STIM technique proved to be suitable for the morphological study of micro-structures fabricated by ion microbeam on polymeric foils. The best configuration is the on-axis geometry using He+2 beam, which provides the best energy resolution without compromising the spatial resolution. In comparison with SEM measurements, advantages such as speed of measurement, non-destructiveness and power of resolving buried structures of the sample were observed. Concerning other features, the similarities between the two techniques make STIM an excellent alternative for morphological analysis of polymeric foils.

As técnicas baseadas em microfeixe de íons energéticos possuem aplicações em diferentes áreas de conhecimento, tais como materiais, biologia, neurociência, ciências ambientais, entre outros. A microssonda iônica também possui a capacidade de fabricar micro-estruturas com aplicações em biomateriais, micro-fotônica, micro-fluídica, membranas porosas e diversas outras. O Laboratório de Implantação Iônica do Instituto de Física (LII) da UFRGS (Universidade Federal do Rio Grande do Sul) conta com uma linha de microssonda iônica que dispõe de detectores para a realização de diversos tipos de experimentos: Emissão de RaiosX Induzida por Partículas (Particle Induced X-Ray Emission – PIXE), Espectroscopia de Espalhamento de Rutherford (Rutherford Backskattering Spectroscopy – RBS), Reação Nuclear (Nuclear Reaction Analysis – NRA) e Microscopia Iônica de Varredura por Transmissão (Scanning Transmission Ion Microscopy – STIM). Além disso, tem-se aperfeiçoado a técnica de micro-fabricação denominada Litografia por Feixe de Prótons (Proton Beam Writing – PBW), atualmente em substratos poliméricos. Este trabalho apresenta dois focos: um deles é a análise de fígado de peixe pela técnica de micro-PIXE. Estes fígados apresentam lesões associadas à exposição de contaminantes. Essas lesões são acompanhadas de células pigmentosas à base de metais chamadas de centros de melanomacrófagos. Observou-se que o fígado de peixe é uma amostra em potencial para estudos ambientais. Outro foco foi o estudo da técnica de STIM em películas poliméricas micro-fabricadas por PBW. Foram estudadas diferentes configurações geométricas para a técnica de STIM, bem como a utilização de feixes de H+1, He+2 e Li+3. Diferentes amostras foram preparadas, desde estruturas simples (possuindo apenas orifícios) até amostras multi-estruturadas do tipo degrau. As medidas de STIM foram comparadas com medidas de Microscopia Eletrônica de Varredura (MEV) de maneira a confirmar a viabilidade e confiabilidade dos estudos morfológicos das amostras em questão. A técnica de STIM se apresentou viável para o estudo morfológico de microestruturas fabricadas por microfeixe de íons em películas poliméricas, sendo que a melhor configuração é na geometria on-axis utilizando o íon He+2 que provê melhor resolução em energia, sem comprometer a resolução espacial. Em comparação com medidas de MEV, apresenta vantagens como maior rapidez, preservação da amostra e o poder de resolver estruturas enterradas na amostra. A similaridade entre as duas técnicas com relação às demais características faz do STIM uma excelente alternativa à mais comumente utilizada para diversas análises morfológicas de películas poliméricas. / Ion beam techniques have many applications in different areas such as materials, biology, neuroscience and environmental science among others. The ion microprobe has also the ability of fabricating microstructures with applications in the fields of biomaterials, micro-photonics, micro-fluidics, porous membranes and many others. The Ion Implantation Laboratory of the Physics Institute at UFRGS has a microprobe beamline equiped with detectors used in different techniques: Particle Induced X-Ray Emission (PIXE), Rutherford Backscattering Spectroscopy (RBS), Nuclear Reaction Analysis (NRA) and Scanning Transmission Ion Microscopy (STIM). Besides, its scanning capability makes allows the use of the micro-fabrication technique called Proton Beam Writing (PBW). Currently, this technique has been used to write micro-structures over polymers. Several projects are in progress for the development and improvement of such techniques, since the microbeam line is a recent acquisition by the laboratory. This work presents two guidelines: one of them is the analysis of fish liver through the micro-PIXE technique. These livers present injuries associated to the exposure of contaminants. The injuries are characterized by the presence of pigmented cells based on metals called melanomacrophage centers. It was observed that the fish liver is a promising sample for environmental studies. The other guideline is the study of the STIM technique on micro-fabricated polymer foils by PBW. Different geometric configurations for the STIM technique were explored, as well as the use of different ion beams (H+1, He+2 and Li+3). Different samples were prepared, ranging from simple structures (containing only one hole) to step-like multi-structured samples. STIM measurements were compared with Scanning Electron Microscopy (SEM) in order to confirm the viability and reliability of morphological studies of the samples under study. The STIM technique proved to be suitable for the morphological study of micro-structures fabricated by ion microbeam on polymeric foils. The best configuration is the on-axis geometry using He+2 beam, which provides the best energy resolution without compromising the spatial resolution. In comparison with SEM measurements, advantages such as speed of measurement, non-destructiveness and power of resolving buried structures of the sample were observed. Concerning other features, the similarities between the two techniques make STIM an excellent alternative for morphological analysis of polymeric foils.

Neste trabalho, o processo de estruturação por microfeixe de íons foi utilizado para a produção de membranas microporosas em folhas comerciais de PET. O processo de estruturação por microfeixe de íons consiste basicamente na interação direta entre um feixe de íons de dimensões micrométricas com o material em questão. As zonas modificadas pelo feixe são removidas do restante do material através de um processo químico. Nesse contexto, durante o desenvolvimento deste trabalho, sistemáticas experimentais para o processo de estruturação foram desenvolvidas. Com o intuito de operacionalizar a linha de microfeixe presente no Laboratório de Implantação Iônica da UFRGS, foi necessário realizar um estudo aprofundado sobre o funcionamento de todo o sistema, verificando problemas e explorando a potencialidade de técnicas não convencionais de análise de materiais. O estudo sobre a sistemática de estruturação ocorreu através da investigação de parâmetros clássicos, como fluência utilizada durante a irradiação e tempo de ataque químico. Para atingir tais objetivos, amostras de poli (tereftalato de etileno) (Mylar) de 12 μm foram irradiadas com microfeixe de íons (H+ e He++) com energias de 3 e 2,2 MeV e fluências que variaram entre 1 x 1011 e 6 x 1015 íons/cm2. Posteriormente à irradiação, as amostras foram submetidas a um ataque químico com solução alcalina de hidróxido de sódio 6 M durante tempos que variaram de 0,5 à 60 minutos. A temperatura do ataque em todos os casos se manteve fixa em 60°C. A caracterização das amostras foi realizada através de microscopia eletrônica de varredura (MEV) e por microscopia de transmissão iônica (STIM). As amostras também foram caracterizadas através de medidas elétricas utilizando um circuito de corrente alternada. O processo de enxertia (grafting) foi testado nas membranas estruturadas, utilizando um hidrogel de PNIPAAm com concentrações de 0,340, 0,450 e 0,700 g/L. Tais resultados também foram analisados através de MEV. O estudo sobre a linha de microfeixe permitiu verificar a existência de problemas relacionados ao registro da carga elétrica durante as irradiações. Além disso, para o ajuste do foco do feixe de íons, foram feitas curvas de calibração de corrente para as lentes magnéticas considerando diferentes energias de feixe. O processo de estruturação através da técnica de microfeixe de íons se mostrou eficaz para a produção de estruturas regulares e definidas em folhas de PET. A fluência ótima de prótons a ser utilizada nos processos de estruturação foi estimada em 6 x 1014 íons/cm2. Para esta fluência, tempos de ataque químico inferiores a 1 minuto já são suficientes para corroer toda a parte irradiada. Entretanto, tempos um pouco mais longos (e.g. 2 minutos) tornam o processo mais reprodutível. Com relação à geometria das estruturas fabricadas, observou-se irregularidades em estruturas que, em princípio, deveriam ser simétricas. Esse problema foi atribuído à assimetria do feixe, proveniente de ajustes dos parâmetros de colimação do feixe. Finalmente, o estudo do processo de enxertia mostrou que o hidrogel adere nas paredes das estruturas, porém não as preenche. Para concentrações elevadas (e.g. 0,7 g/L) o processo não é tão eficiente, sendo que não é verificada a redução da área das microestruturas pela inserção do hidrogel. As medidas elétricas mostraram a existência de regimes distintos e dependentes da frequência da corrente alternada. Os polímeros apresentaram basicamente comportamentos resistivo e capacitivo. / In this work, the process of irradiation of PET foils with ion beams in the micrometer size range was used for the production of microporous membranes. Basically, this process consists on the direct interaction between the ion beam and the material under study. The regions modified by the beam are removed from the material through a chemical process. In this context, experimental procedures for the production process of the membranes were developed during the course of this work. In order to make the microbeam station of the Ion Implantation Laboratory of the Federal University of Rio Grande do Sul (UFRGS), it was necessary to perform a thorough study of the operational parameters of the system, thus allowing a proper identification of problems and providing grounds for pushing the technique to the frontier of materials science. To achieve such objectives, foils of polyethylene terephtalate (Mylar®) 12 μm thick were irradiated with H+ and He++ ions with 3 e 2,2 MeV respectively. Fluencies varied from 1 x 1011 and 6 x 1015 ions/cm2. After the irradiation, the foils were submitted to an etching procedure using alkaline solution of sodium hydroxide at 6 M during periods of time varying from 0,5 to 60 minutes. In all cases, the temperature of the etching was fixed at 60°C. The characterization of the samples was performed through scanning electron microscopy (SEM) and scanning transmission ion microscopy (STIM). The samples also were characterized by electric measurements using an AC current circuit. The process of grafting was tested on the structured membranes using a PNIPAAm hydrogel with concentrations of 0,340, 0,450 and 0,700 g/L. The results of this study were also analyzed through MEV. With the present study, it was possible to pinpoint problems related to the integration and recording of the charge during the irradiations. Besides that, calibration curves were obtained relating the electric currents needed on the magnetic lenses for an optimal ion beam focus and the beam energy. The irradiation process with ion beam proved itself efficient for the production of regular patterns on PET foils. The optimum dose of prótons to be used on the patterning processes was estimated in 6 x 1014 ions/cm2. For this dose, etching times smaller than 1 minute were enough to remove all the irradiated area. However, times slightly longer (e.g. 2 minutes) make the process more reproducible. Regarding the geometry of the patterns generated by the ion irradiation, asymmetries were observed on structures that were supposed to be symmetric. This problem was attributed to the asymmetry of the beam spot on the target due to the settings of the objective slits that collimates the beam. The study of the grafting process showed that the hydrogel adheres to the structures walls, but does not fill it. For high concentrations (e.g. 0,7 g/L), the process is not efficient, since no reduction of the area of the microstructures by the insertion of the hydrogel was observed. The electric measurements showed the existence of distinct regimes as a function of the frequency of the alternate current. Basically, the polymer foils present resistive and capacitive behaviors.

Denna uppsats problematiserar hur kollektiva förvaltningsorganisationer fördelar pengar insamlade från offentligt framförd klubbmusik samt undersöker hur etablerade dj’s och producenter ser på dagens system och tror sig påverkas av detta. Med djupintervjuer av fyra personer undersöker uppsatsen även hur dessa ser på rapportering av den spelade musiken, vilka problem som kan uppstå och om det finns tekniska möjligheter att underlätta för korrekt fördelning. Resultaten visar att intervjupersonerna skiljer sig åt i vilken typ av karriär de har och därmed hur man förhåller sig till dagens system. Ingen ställde sig dock positiv till att rapportera in spellistor med vad de spelat som dj’s, man lyfte även fram problemet med att ersätta musik som spelas men ännu inte släppts officiellt. Det framkom också att vi idag har tekniska möjligheter att övervaka vad som spelas genom användandet av akustiska fingeravtryck. Jag diskuterade att det är först när ett sådant system är på plats som vi kan se hur mycket fel dagens pengar fördelas.

Une hyperactivation de la prolifération des cellules musculaires lisses vasculaires (CMLV) contribue à la pathogenèse des maladies des vaisseaux. Des travaux antérieurs suggèrent que l’augmentation de l'adénosine monophosphate cyclique (AMPc) inhibe la prolifération des CMLV. Provoquer une augmentation d’AMPc préviendrait aussi certaines maladies vasculaires qui sont associées à des altérations dans sa signalisation impliquant l'activité de la protéine kinase A (PKA). Des études ont démontré la contribution du facteur de transcription « early growth response protein-1» (Egr-1) dans la pathogenèse des maladies vasculaires et une surexpression d’Egr-1 a été rapportée dans des modèles d'athérosclérose et d'hyperplasie intimale. Divers agents vasoactifs contrôlent l'expression d’Egr-1 suivant des mécanismes qui ont fait l’objet de plusieurs études mais demeurent incomplètement élucidés. L'angiotensine-II (Ang-II) est l'un des principaux peptides vasoactifs impliqués dans la pathogenèse des maladies vasculaires. Une des voies de signalisation induite par l’Ang-II implique l’augmentation du calcium (Ca2+) intracellulaire. Celle-ci se produit par l’activation de l'entrée de calcium opérée par la relâche des réserves (SOCE) de Ca2+ réticulaire suite à l’activation du récepteur à l’inositol-3-phosphate (IP3R) et le recrutement ultérieur du complexe conducteur formé par la molécule d'interaction stromale 1 (STIM-1) et le canal Orai-1. Bien qu’il ait déjà été démontré que l’expression de l'Egr-1 est régulée par la signalisation calcique en réponse à plusieurs stimuli, l'implication du complexe STIM-1/Orai-1 dans l'expression d'Egr-1 dans la CMLV n’a jamais été étudiée. De même, la question de savoir si la signalisation induite par l'Ang-II conduisant à l'expression d'Egr-1 est modulée par l'AMPc n’a jamais été explorée. Par conséquent, les travaux menés dans cette thèse ont consisté à examiner le rôle de la signalisation du Ca2+ dans l'expression d'Egr-1 induite par l’Ang-II dans la CMLV avec une attention particulière portée sur le rôle joué par STIM-1 et Orai-1. En outre, nous avons examiné l'effet de l’augmentation de l’AMPc sur l'expression d'Egr-1 induite par l’Ang-II et étudié les voies de signalisation associées. Nos données montrent que l’inhibition du récepteur IP3R et du SOCE par le 2-aminoéthoxydiphénylborate atténue la libération de Ca2+ induite par l’Ang-II et ceci s’accompagne d’une baisse des niveaux d’expression de protéine et d’ARN messager de l’Egr-1. La stimulation de l’expression de l'Egr-1 a également été supprimée à la suite du blocage de la calmoduline et de la protéine kinase CaMKII. De plus, le blocage par interférence d’ARN de l’expression de STIM-1 et Orai-1 a atténué l'expression d'Egr-1 induite par l’Ang-II ainsi que la phosphorylation des protéines ERK et CREB. Par ailleurs, l'isoproterenol (ISO) et la forskoline (FSK), deux activateurs de l'adénylate cyclase ont atténué de manière dose-dépendante l'expression d'Egr-1 induite par l’Ang-II. Des réponses similaires ont été observées en utilisant des analogues non spécifique (dibutyryl-cAMP) et PKA-spécifique (Benzoyl-cAMP) de l’AMPc, ainsi qu'un inhibiteur à large spectre de l'activité phosphodiesterase intracellualaire (isobutylméthylxanthine). L'inhibition de l'expression d'Egr-1 induite par l’Ang-II s’accompagne d'une augmentation de l’activité de la PKA mesurée par la phosphorylation de la « phosphoprotéine activée par les vasodilatateurs (VASP) », et d’une diminution concomitante de la phosphorylation de la protéine ERK. Le blocage pharmacologique de la PKA a réduit la phosphorylation de VASP et restauré la phosphorylation de la protéine ERK ainsi que l'expression d'Egr-1 en présence de l’Ang-II. En résumé, nos données démontrent que la voie STIM-1/Orai-1 /Ca2+ médie l'expression de l'Egr-1 induite par l'Ang-II dans la CMLV et suggèrent que la suppression de la réponse à l’Ang-II menant à l’expression de l'Egr-1 peut expliquer les effets vasoprotecteurs de l’AMPc. En outre, ces travaux montrent que les mécanismes moléculaires de régulation de l’expression d’Egr-1 en réponse aux signaux externes culminent vers la modulation des cascades de signalisation en aval de la protéine ERK dans les CMLV. / Aberrant vascular smooth muscle cell (VSMC) proliferative responses contribute to the development of neointimal lesions. Cyclic adenosine monophosphate (cAMP) is believed to inhibit VSMC proliferation, and vascular diseases are associated with impairments in cAMP-induced signalling responses involving protein kinase A (PKA) signaling. An enhanced expression of the early growth response protein-1 (Egr-1), a zinc finger transcription factor, has been reported in models of vascular diseases and, a crucial role of Egr-1 in regulating the expression of genes implicated in neointimal formation leading to atherogenesis has been suggested. Various vasoactive factors have been shown to modulate Egr-1 expression in VSMC via mechanisms which remain to be completely understood. Angiotensin-II (Ang-II) is one of the key vasoactive peptides implicated in the pathogenesis of vascular diseases. Ang-II elevates intracellular calcium (Ca2+) through activation of voltage-gated calcium channels as well as store-operated calcium channels. The store-operated calcium entry (SOCE) involves an inositol-3-phosphate receptor (IP3R)-coupled depletion of endoplasmic reticular Ca2+ and a subsequent activation of the stromal interaction molecule 1 (STIM-1) /Orai-1 complex. Although Egr-1 has been demonstrated to be upregulated in a Ca2+-dependent fashion in response to several stimuli, the involvement of STIM-1/Orai-1-dependent signaling in Egr-1 expression in VSMC has never been addressed. Besides, whether Ang-II-induced signaling leading to Egr-1 expression is modulated by cAMP-dependent signaling pathway remains unexplored. Therefore, in the present studies, we have examined the role of Ca2+ signaling in Ang-II-induced Egr-1 expression in VSMC and investigated the contribution of STIM-1 or Orai-1. Additionnaly, we have examined the effect of cAMP on Ang-II-induced expression of Egr-1 and have investigated the associated signalling pathways. Pharmacological blockade of IP3R and SOCE by 2-aminoethoxydiphenylborate (2-APB) decreased Ang-II-induced Ca2+ release and attenuated Ang-II-induced enhanced expression of Egr-1 protein and mRNA levels. Egr-1 upregulation was also suppressed following blockade of calmodulin and CaMKII. Furthermore, RNA interference-mediated depletion of STIM-1 or Orai-1 attenuated Ang-II-induced Egr-1 expression, as well as Ang-II-induced phosphorylation of ERK1/2 and CREB. Moreover, isoproterenol (ISO) and forskolin (FSK), two respective receptor and non-receptor activators of adenylate cyclase, attenuated Ang-II-induced Egr-1 expression in a dose-dependent fashion. Similar responses were observed using non-specific (dibutyryl-cAMP) and PKA-specific (Benzoyl-cAMP) analogs of cAMP, as well as a broad spectrum inhibitor of intracellular phosphodiesterase activity (isobutylmethylxanthine). The inhibition of Ang-II-induced Egr-1 expression was accompanied by an increase in serine 157 phosphorylation of the vasodilator-activated phosphoprotein (VASP), a marker of PKA activity, and this was associated with a concomitant decrease in ERK phosphorylation. Pharmacological blockade of PKA using H89 decreased VASP phosphorylation, restored Ang-II-induced ERK phosphorylation and abolished ISO- and FSK-mediated inhibition of Ang-II-induced Egr-1 expression. In summary, our data demonstrate that STIM-1/Orai-1/Ca2+-dependent signaling pathways mediate Ang-II-induced Egr-1 expression in A-10 VSMC and suggest that PKA-mediated suppression of Ang-II-induced Egr-1 expression and phosphorylation of ERK may be among the mechanisms by which cAMP exerts its vasoprotective effects. In addition, our data supports the notion that stimuli-induced regulation of Egr-1 expression involves the participation of signaling cascades downstream of ERK in VSMC.

No description available.

<p>Recent development of the internet encompasses elements such as social networks, blogs and wikis. With the help of these elements, popularly gathered under the umbrella term web 2.0, the pirates supporting file sharing have made the file sharing debate a national concern in Sweden.</p><p>This thesis studies the pirates’ and anti-pirates’ use of PR by qualitative case studies of organizations representing the two different sides in the debate. The use of PR by the different organizations is compared, the differences and similarities are considered and characteristics of the new PR are brought to attention.</p><p>The study finds that PR is taking a new direction and that these new ways to use PR are important to achieve success in influencing opinion. The two sides use PR in very different ways where the anti-pirates use more traditional ways of PR while the pirates make use of new PR methods virtually exclusively.</p>

Recent development of the internet encompasses elements such as social networks, blogs and wikis. With the help of these elements, popularly gathered under the umbrella term web 2.0, the pirates supporting file sharing have made the file sharing debate a national concern in Sweden. This thesis studies the pirates’ and anti-pirates’ use of PR by qualitative case studies of organizations representing the two different sides in the debate. The use of PR by the different organizations is compared, the differences and similarities are considered and characteristics of the new PR are brought to attention. The study finds that PR is taking a new direction and that these new ways to use PR are important to achieve success in influencing opinion. The two sides use PR in very different ways where the anti-pirates use more traditional ways of PR while the pirates make use of new PR methods virtually exclusively.

Syftet med det examensarbete som genomförts har varit att kartlägga processen för att på egen hand lansera sin musik i form av ett fullängdsalbum tillgängligt att köpas både i form av digitala nedladdningar och som traditionell CD-skiva. I arbetet har vi bekantat oss med viktiga aspekter angående distributionsavtal, upphovsrätt, marknadsföring, design, foto, logotyper, texter och koder man bör känna till. En stor del av arbetet har även ägnats åt att förklara och utreda den sista delen i ljudbearbetningen - mastring. Dessutom har vi satsat på marknadsförning och exponering på Internet samt olika betalningslösningar som möjliggör skivans försäljning. För att erhålla de nödvändiga delarna i arbetsprocessen har kontakt upprättats med bland andra: IFPI, Grammotex, N©B och Dicentia. I rapporten får vi följa den progressiva rockgruppen Cap Outrun och deras väg till målet i form av en fallstudie.

Les polynucléaires neutrophiles représentent 50-70% des leucocytes sanguins et possèdent un rôle majeur dans la défense de l'organisme contre les pathogènes. Le Ca2+ est un second messager qui joue un rôle primordial dans le chimiotactisme, la phagocytose, la dégranulation et la production de formes réactives de l'oxygène (FRO) afin de neutraliser l'agent pathogène. Dans ces cellules, l'influx calcique de type SOCE est essentiel pour l'homéostasie calcique. Il est peu étudié en raison du manque d'outils pharmacologiques spécifiques d'où l'importance dans un premier temps de chercher de nouvelles molécules. Les cellules T Jurkat dont le SOCE est largement caractérisé servent de modèle pour la caractérisation initiale de ces molécules. Le 2-APB est parmi les molécules les plus largement utilisées dans la caractérisation du SOCE en raison de sa double activité sur le SOCE avec une potentialisation à [1-10 μM] et une inhibition à [> 20 μM]. En revanche, ce produit manque de spécificité et agit sur d'autres cibles cellulaires comme les récepteurs à l'inositol (1,4,5)-trisphosphate (InsP3Rs). La 1ère étape est de sélectionner à partir d'analogues commerciaux du 2-APB (Methoxy-APB, Dimethoxy-APB, Cyclic-APB, Benzothienyl-APB, Thienyl-APB et MDEB), des composés plus spécifiques et également plus efficaces que la molécule mère. Deux molécules se sont distinguées : le MDEB comme uniquement potentialisant du SOCE et le Benzothienyl-APB comme un puissant inhibiteur. En revanche, tous les analogues du 2-APB inhibent les InsP3Rs à l'exception du MDEB qui semble plus spécifique du SOCE. L'effet du MDEB sur le courant calcique, ICRAC, a été étudié grâce à la technique du patch-clamp. Il augmente d'environ 4 fois l'amplitude de ICRAC par rapport à celle enregistrée dans les cellules contrôle. Par ailleurs, le MDEB ralentie l'inactivation rapide de ICRAC due au Ca2+. Sur le plan physiologique, le MDEB à des concentrations croissantes inhibe la synthèse de l'IL-2 dans les cellules Jurkat stimulées et ceci malgré son effet potentialisant du SOCE. Cette activité est liée à son effet pro-apoptotique dans les cellules Jurkat stimulées. Le MDEB et le Benzothienyl-APB caractérisés dans la 1ère partie nous ont servi d'outils potentiels afin d'étudier le SOCE des cellules PLB-985 différenciées en cellules proches de neutrophiles. Le SOCE a été induit soit par un traitement des cellules avec la thapsigargine (Tg) soit de manière physiologique avec les peptides fMLF et le WKYMVm deux chimioattractants, ligands des récepteurs aux peptides formylés FPR et FPRL1 respectivement. En plus, le SOCE induit par la Tg est modulable par le 2-APB, potentialisé par le MDEB et inhibé par le Benzothienyl-APB. La phagocytose des levures par les cellules PLB-985 différenciées ainsi que la production de FRO intraphagosomales ont été inhibées par le MDEB et le Benzothienyl-APB. Les FRO extracellulaires ont été également inhibées par Benzothienyl-APB en revanche à cause de la forte interférence du MDEB avec la technique de mesure nous n'avons pas pu étudier ses activités. En conclusion, le MDEB et le Benzothienyl-APB sont de nouveaux outils pharmacologiques potentialisant ou inhibant le SOCE des leucocytes, qui nous permettront dans l'avenir une meilleure compréhension de l'entrée calcique et ses rôles dans ces cellules.