Glucose is a monosaccharide and fuel for body, it cannot pass through membrane by simple diffusion so, integral transmembrane proteins named glucose transporters (Gluts) are involved in the regulation of the movement of glucose between the extracellular and intracellular spaces within the body. GLUT1 and GLUT3 have previously been shown by cold detergent extraction methods to reside in distinct plasma membrane domains in non-polarized mammalian cells, with GLUT1, but not GLUT3, residing  in detergent-resistant membrane (DRM) domains. To confirm this observation under less invasive conditions, molecular fusion tags are inserted in the first external loop in Glut1 with biotin ligase acceptor peptide (BLAP) between Ser-55 and Ile-56 and in Glut3 with Acyl carrier peptide (ACP) in between Val-57 and Leu-58 respectively. These Glut fusion proteins will be used in order to confirm these observations by fluorescence recovery after photobleaching (FRAP) and single molecule fluorescence microscopy in live cells. hGLUT1-EGFP, hGLUT1 (AgeI)-EGFP recombinants were constructed and transfected human embryonic kidney cells (HEK-293) quantum dot images supports the fact that EGFP transfected cells uniformly and is distributed in the cell cytoplasm, hGLUT1-EGFP transfected cells and is localized to the cell membrane and hGLUT1 (AgeI)-EGFP transfected cells and located to the plasma membrane with high intensity.

<p>Glucose is a monosaccharide and fuel for body, it cannot pass through membrane by simple diffusion so, integral transmembrane proteins named glucose transporters (Gluts) are involved in the regulation of the movement of glucose between the extracellular and intracellular spaces within the body. GLUT1 and GLUT3 have previously been shown by cold detergent extraction methods to reside in distinct plasma membrane domains in non-polarized mammalian cells, with GLUT1, but not GLUT3, residing  in detergent-resistant membrane (DRM) domains. To confirm this observation under less invasive conditions, molecular fusion tags are inserted in the first external loop in Glut1 with biotin ligase acceptor peptide (BLAP) between Ser-55 and Ile-56 and in Glut3 with Acyl carrier peptide (ACP) in between Val-57 and Leu-58 respectively. These Glut fusion proteins will be used in order to confirm these observations by fluorescence recovery after photobleaching (FRAP) and single molecule fluorescence microscopy in live cells. hGLUT1-EGFP, hGLUT1 (<em>AgeI</em>)-EGFP recombinants were constructed and transfected human embryonic kidney cells (HEK-293) quantum dot images supports the fact that EGFP transfected cells uniformly and is distributed in the cell cytoplasm, hGLUT1-EGFP transfected cells and is localized to the cell membrane and hGLUT1 (<em>AgeI</em>)-EGFP transfected cells and located to the plasma membrane with high intensity.</p>

<p>Abstract</p><p>G-protein coupled receptors (GPCRs) are having the high medical importance since almost half of the medicinal drugs are designed as modulators of receptor molecules. Crystal structure or NMR structures of GPCRs are very difficult to determine because all GPCRs are typically bound to the cell membrane and thus their molecular activation mechanism is still unclear. The recent publication of the crystal structure of the 2-adrenoreceptor will provide new insights in the field of GPCR research.</p><p>Ghrelin is a peptide growth hormone which binds to the growth hormone secretagogue receptor (GHS-R) and stimulates the release of growth hormone. Based on the known ghrelin receptor binding core sequences wFwLL (upper letter and lower letter representative for L-form and D-form of the amino acids respectively), we prepared two novel peptide analogs with terminal S-(2-aminoethylsulfenyl) cysteine residues. These peptides were tested for their ability to suppress the binding of ghrelin to transfected COS7 cell-line (Kidney fibroblast line from the green African monkey) cells expressing the ghrelin wild-type receptor or certain mutants thereof. As a result we observed a significant reduction of the total number of binding sites accessible for ghrelin, which increased with the time the cells were incubated with our test compounds. This observations support our hypothesis that the peptides we tested form a covalent bond with free thiols located closely to the ligand binding-site of the receptor protein by disulfide thiol exchange which is an interesting target for development of anti-obesity drugs.</p>

Abstract G-protein coupled receptors (GPCRs) are having the high medical importance since almost half of the medicinal drugs are designed as modulators of receptor molecules. Crystal structure or NMR structures of GPCRs are very difficult to determine because all GPCRs are typically bound to the cell membrane and thus their molecular activation mechanism is still unclear. The recent publication of the crystal structure of the 2-adrenoreceptor will provide new insights in the field of GPCR research. Ghrelin is a peptide growth hormone which binds to the growth hormone secretagogue receptor (GHS-R) and stimulates the release of growth hormone. Based on the known ghrelin receptor binding core sequences wFwLL (upper letter and lower letter representative for L-form and D-form of the amino acids respectively), we prepared two novel peptide analogs with terminal S-(2-aminoethylsulfenyl) cysteine residues. These peptides were tested for their ability to suppress the binding of ghrelin to transfected COS7 cell-line (Kidney fibroblast line from the green African monkey) cells expressing the ghrelin wild-type receptor or certain mutants thereof. As a result we observed a significant reduction of the total number of binding sites accessible for ghrelin, which increased with the time the cells were incubated with our test compounds. This observations support our hypothesis that the peptides we tested form a covalent bond with free thiols located closely to the ligand binding-site of the receptor protein by disulfide thiol exchange which is an interesting target for development of anti-obesity drugs.

Detta examensarbete är skrivet som en del i programmet industriell organisation och ekonomi, med inriktning logistik och ledning vid Tekniska Högskolan i Jönköping. Rapporten är skriven på uppdrag av Skövde flygplatsbolag AB, som är ett kommunalt flygplatsbolag. Skövde flygplats är belägen ett par mil utanför Skövde centrum. Flygplatsen trafikeras idag mestadels av transportflyg men även enstaka mindre passagerarflyg. Idag är Kinnarps, Volvo Powertrain, Räddningstjänsten och klubbverksamhet deras största aktörer.   Syftet med examensarbetet har varit att beskriva Skövde flygplats nuvarande verksamhet, samt att utreda flygplatsens miljöledningssystem (MLS). Flygplatsen har även efterfrågat en översyn av det nuvarande miljöledningssystemet på Skövdes flygplats. Rapporten avspegla därför den genomförda undersökningen, men även ge förslag på förbättringar som kan göras med avseende på flygplatsens miljö och verksamhet.   För att uppfylla examensarbetets syfte, har ett antal delmål skapats. Delmålen är till för att förbättra rapportens resultat och syfte. I studiens teoriavsnitt beskrivs därför vilka delar som ingår i ett MLS, standarder och förenklade MLS. Studien har även kartlagt Skövde flygplats verksamhet, eftersom verksamheten idag är otydlig och ofullständig.   Eftersom flygplatsens verksamhet idag saknar information och dokumentation kring flygplatsen, har ett antal studiebesök genomförts. Under studiebesöken har studenterna observerat och intervjuat personal, samt fått ta del av dokumentation kring flygplatsen. Därför har studien valt att genomföra en dokumentstudie. Resultatet av dokumentstudien redovisas sedan i rapportens kapitel 4, ”Nulägesbeskrivning”, 5 ”Nulägesanalys”.   Under arbetets gång har ett antal brister påträffats på Skövde flygplats. Idag saknar till exempel flygplatsen information och dokumentation om sin verksamhet och MLS. Flygplatsen är till exempel idag miljödiplomerad, men saknar samtidigt stöd från studiens teori att få kalla sig miljödiplomerad. Verksamheten är idag komplex och det finns många olika uppfattningar kring flygplatsens verksamhet. Resultatet är baserat på examensarbetets dokumentstudie tillsammans med studiens teoriunderlag. Resultatet presenteras i det avslutande kapitlet i form av förslag och rekommendationer till Skövde flygplats. / This report was written as a part of the bachelor program, Industrial Organization and Economics, focusing on logistics and management, at the School of Engineering in Jonkoping.  The report was written on behalf of Skövde Airport Company, which is a municipally owned airport company.  Skövde Airport is located within a few miles outside the city of Skövde, Sweden.   The airport serves mainly cargo planes but occasionally serves smaller passenger flights as well.  Today, companies like Kinnarps AB and Volvo Powertrain, are their biggest customers.  The airport also includes a fire station service and offices for these larger customers.   The purpose of this study is to provide an assessment of Skövde Airport’s surrounding issues. Our goal is to provide suggestions as to what the airport can implement as well as continue working on to improve the airport environment.  Therefore the study also examines other options in terms of ways to prepare to work with the environment; these systems are called “Environment Management Systems (EMS).”  Consequently, the study has taken parts of these EMS: s to identify and describe what parts are most critical from an environmental perspective.   The report includes a critical examination of the airport and explanation on how the airport works with the environment. The investigation portion of this thesis presents our mapping of the Skövde Airport Company’s current EMS.  The analysis describes everyday business activities and how these everyday activities are impacting the environment. Finally, the report concludes our reflection of the outcome of implementing these suggestions we have presented.   During this process the study found a number of deficiencies at Skövde airport. Today Skövde airport is missing vital information and documentation on its activities and the EMS. The airport currently holds a diploma for their EMS, but according to this study it’s missing vital information and documentation. Skövde airport is a complex organization to describe and even today there are many different opinions and views about its function and business.   The result of this study is therefore based on a document study. During the project time information had been collected from documentation, visits at the airport, interviews and communication with employees.  The result is presented in the final chapter in the form of proposals and recommendations to Skövde Airport.