Differential Regulation of c-FLIP Isoforms Through Post-translational Modifications

Cells are constantly responding to signals from the surrounding tissues and the environment. To dispose of infected and potentially dangerous cells, to ensure the optimal execution of developmental processes and to maintain tissue homeostasis, a multicellular organism needs to tightly control both the number and the quality of its cells. Apoptosis is a form of active cellular self-destruction that enables an organism to regulate its cell number by deleting damaged or potentially dangerous cells. Apoptosis can be induced by death ligands, which bind to death receptors on the cell surface. Ligation of the receptors leads to the formation of an intracellular death inducing signaling complex (DISC). One of the DISC components is caspase-8, a protease that triggers the caspase cascade and is thereby a key initiator of programmed cell death. The activation of caspase-8 is controlled by the cellular FLICE-inhibitory proteins (c-FLIPs). Consequently, sensitivity towards receptor-mediated apoptosis is determined by the amount of c-FLIP, and the c-FLIP levels are actively regulated for example during erythroid differentiation of K562 erythroleukemia cells and by hyperthermia in Jurkat leukemia cells. The aim of my thesis was to investigate how c-FLIP is regulated during these processes. We found that c-FLIP isoforms are short-lived proteins, although c-FLIPS had an even shorter half-life than c-FLIPL. In both experimental models, increased death receptor sensitivity correlated with induced ubiquitylation and consequent proteasomal degradation of c-FLIP. Furthermore, we elucidated how phosphorylation regulates the biological functions and the turnover of c-FLIP, thereby contributing to death receptor sensitivity. We mapped the first phosphorylation sites on c-FLIP and dissected how their phosphorylation affects c-FLIP. Moreover, we demonstrated that phosphorylation of serine 193, a phosphorylated residue common to all c-FLIPs, is primarily mediated by the classical PKC. Furthermore, we discovered a novel connection between the phosphorylation and ubiquitylation of c-FLIP: phosphorylation of S193 protects c-FLIP from ubiquitylation. Surprisingly, although all c-FLIP isoforms are phosphorylated on this conserved residue, the biological outcome is different for the long and short isoforms, since S193 specifically prolongs the half-lives of the short c-FLIP isoforms, but not c-FLIPL. To summarize, we show that c-FLIP proteins are modified by ubiquitylation and phosphorylation, and that the biological outcomes of these modifications are isoform-specifically determined.

Om nyttan och nödwändigheten för en präst, at äga insikt i medicine

Painovuosi nimekkeestä.

Informationsbeteende och inlärningsmiljöer : en kvalitativ studie av medicine studerandes informationsbeteende i två olika utbildningsprogram

Avhandlingens övergripande syfte är att granska relationerna mellan olika undervisningsmetoder och studenters informationsbeteende, vilket i denna undersökning inbegriper även deras informationskompetens. Vikten av att undersöka dessa förhållanden kan motiveras med att både kunskap om de faktorer som påverkar utvecklandet av informationskompetens och forskning som tar fram olika mönster i studenternas informationsbeteende behövs för att sådana inlärningsmiljöer, informationssystem och -tjänster som stöder studenternas inlärning skall kunna utvecklas. I avhandlingen söks svar på följande frågor: 1. Vilka faktorer i inlärningsmiljöerna, dvs. problembaserad inlärningsmiljö (pbl) och traditionell inlärningsmiljö, påverkar informationsbeteendet och hur påverkar dessa faktorer? 2. Hurdan information behövs i inlärningsprocessen? Hur anskaffas informationen? Vilka informationskanaler och -källor används och hur används de? 3. Hur används information i samband med inlärningen? I undersökningen används en kvalitativ forskningsansats och det huvudsakliga undersökningsmaterialet består av intervjuer med 16 medicine studerande som studerar enligt en problembaserad inlärningsmetod och 15 studerande som studerar i ett traditionellt ämnesbaserat utbildningsprogram. Den empiriska delen av undersökningen utfördes i slutet av 1990-talet. Resultaten indikerar att en problembaserad inlärningsmiljö utvecklar förståelsen av kunskap, aktiverar informationsanskaffningen och informationsanvändningen, samt främjar utvecklingen av studenternas informationskompetens såsom den definierades i denna undersökning. Högre nivå av informationskompetens och aktiv informationsanvändning förekom emellertid i båda utbildningsprogrammen även bland studenter som hade påbörjat de fördjupade studiernas slutarbete, vilket framhäver motivationens och de verkliga informationsbehovens roll i informationsbeteendet och i utvecklandet av informationskompetensen.

Translational research : pharmaceutical industry view

Translationaalinen lääketutkimus ja -kehitys.

Translational models of coronary artery disease, myocardial infarction and heart failure. Development, validation and in vivo imaging studies using positron emission tomography

Coronary artery disease is an atherosclerotic disease, which leads to narrowing of coronary arteries, deteriorated myocardial blood flow and myocardial ischaemia. In acute myocardial infarction, a prolonged period of myocardial ischaemia leads to myocardial necrosis. Necrotic myocardium is replaced with scar tissue. Myocardial infarction results in various changes in cardiac structure and function over time that results in “adverse remodelling”. This remodelling may result in a progressive worsening of cardiac function and development of chronic heart failure. In this thesis, we developed and validated three different large animal models of coronary artery disease, myocardial ischaemia and infarction for translational studies. In the first study the coronary artery disease model had both induced diabetes and hypercholesterolemia. In the second study myocardial ischaemia and infarction were caused by a surgical method and in the third study by catheterisation. For model characterisation, we used non-invasive positron emission tomography (PET) methods for measurement of myocardial perfusion, oxidative metabolism and glucose utilisation. Additionally, cardiac function was measured by echocardiography and computed tomography. To study the metabolic changes that occur during atherosclerosis, a hypercholesterolemic and diabetic model was used with [18F] fluorodeoxyglucose ([18F]FDG) PET-imaging technology. Coronary occlusion models were used to evaluate metabolic and structural changes in the heart and the cardioprotective effects of levosimendan during post-infarction cardiac remodelling. Large animal models were used in testing of novel radiopharmaceuticals for myocardial perfusion imaging. In the coronary artery disease model, we observed atherosclerotic lesions that were associated with focally increased [18F]FDG uptake. In heart failure models, chronic myocardial infarction led to the worsening of systolic function, cardiac remodelling and decreased efficiency of cardiac pumping function. Levosimendan therapy reduced post-infarction myocardial infarct size and improved cardiac function. The novel 68Ga-labeled radiopharmaceuticals tested in this study were not successful for the determination of myocardial blood flow. In conclusion, diabetes and hypercholesterolemia lead to the development of early phase atherosclerotic lesions. Coronary artery occlusion produced considerable myocardial ischaemia and later infarction following myocardial remodelling. The experimental models evaluated in these studies will enable further studies concerning disease mechanisms, new radiopharmaceuticals and interventions in coronary artery disease and heart failure.