PV-1: Leukocyte trafficking molecule and vascular marker

The distinction between lymphatic vessels and blood vessels is a crucial factor in many studies in immunology, vascular biology and cancer biology. They both share several characteristics and perform related, though different functions. They are equally important for the performance of the human immune system with the continuous recirculation of leukocytes from the tissues via lymphatics to the blood vessels and back into the tissue presenting the link between both systems. This study was undertaken to elucidate the differences in the gene expression between primary blood- and lymphatic endothelial cells as well as the two immortalized cell lines HMEC-1 (human microvascular endothelial cell line 1) and TIME (telomerase immortalized microvascular endothelial cell line). Furthermore, we wanted to investigate the mystery surrounding the identity of the antigen recognized by the prototype blood vascular marker PAL-E. In the last step we wanted to study whether the PAL-E antigen would be involved in the process of leukocyte migration from the bloodstream into the surrounding tissue. Our results clearly show that the gene expression in primary blood endothelial cells (BEC), lymphatic endothelial cells (LEC) and the cell lines HMEC-1 and TIME is plastic. Comparison of a large set of BEC- and LEC datasets allowed us to assemble a catalog of new, stable BEC- or LEC specific markers, which we verified in independent experiments. Additionally, several lines of evidence demonstrated that PAL-E recognizes plasmalemma vesicle associated protein 1 (PV-1), which can form complexes with vimentin and neuropilin-1. Finally, numerous in vitro and in vivo experiments identify the first function of the protein PV-1 during leukocyte trafficking, where it acts as regulatory molecule.

Vesicular Trafficking in Osteoclasts

Osteoclasts are multinucleated bone-degrading cells that undergo large changes in their polarisation and vesicular trafficking during the bone resorption cycle. Rab proteins are small GTPases that offer both temporal and spatial regulation to the transport between membranous organelles. Previously the presence and function of only few of the currently known 60 Rab proteins in osteoclasts have been reported. In this study, the expression of 26 Rab genes in bone-resorbing osteoclasts was demonstrated with gene-specific primer pairs. The further analysis of three Rab genes during human osteoclast differentiation revealed that Rab13 gene is highly induced during osteoclastogenesis. The presence of Rab13 protein in the secretory vesicles directed towards the ruffled border and in the endocytotic or transcytotic pathways in resorbing osteoclasts was excluded. The localisation of Rab13 suggests that that it is associated with a previously unknown vesicle population travelling between the trans-Golgi network and the basolateral membrane in bone resorbing osteoclasts. Rab proteins convey their functions by binding to specific effector proteins. We found a novel Rab13 interaction with endospanins-1 and -2 that are yet poorly characterised small transmembrane proteins. The Rab13 subfamily member Rab8 also bound to endospanins, while Rab10 and unrelated Rabs did not. Rab13 and endospanin-2 co-localised in perinuclear vesicles in transfected cells, demonstrating the interaction also in vivo. The inhibition of Rab13 did not interfere with the localisation of endospanin-2 nor did it affect the cell surface expression of growth hormone receptor, as has been previously described for endospanins. The physiological role of this novel protein-protein interaction thus remains to be clarified. The analysis of the transcytotic route in bone resorbing osteoclasts revealed that multiple vesicle populations arise from the ruffled border and transport the bone degradation products for exocytosis. These vesicles are directed to the functional secretory domain that is encircled by an actin-based molecular barrier. Furthermore, the transcytotic vesicles contain abundant Helix pomatia lectin binding sites and represent lipid raft concentrates. Finally, autophagosomal compartments may also be involved in the transcytosis.

Balancing the quotation process by LEAN methods

This thesis studies the possibility to use lean tools and methods in a quotation process which is carried out in an office environment. The aim of the study was to find out and test the relevant lean tools and methods which can help to balance and standardize the quotation process, and reduce the variance in quotation lead times and in quality. Seminal works, researches and guide books related to the topic were used as the basis for the theory development. Based on the literature review and the case company’s own lean experience, the applicable lean tools and methods were selected to be tested by a sales support team. Leveling production, by product categorization and value stream mapping, was a key method to be used to balance the quotation process. 5S method was started concurrently for standardizing the work. Results of the testing period showed that lean tools and methods are applicable in office process and selected tools and methods helped to balance and standardize the quotation process. Case company’s sales support team decided to implement new lean based quotation process model.

Building climate justice : an analysis of how the nexus between climate change and human rights shapes public policy agendas and alternatives

This thesis provides an analysis of how the nexus between climate change and human rights shapes public policy agendas and alternatives. It draws upon seminal work conducted by John Kingdon, whose landmark publication “Agendas, alternatives, and public policy” described how separate streams of problems, solutions, and politics converge to move an issue onto the public policy agenda toward potential government action. Building on Kingdon’s framework, this research explores how human rights contribute to surfacing the problem of climate change; developing alternative approaches to tackling climate change; and improving the political environment necessary for addressing climate change with sufficient ambition. The study reveals that climate change undermines the realization of human rights and that human rights can be effective tools in building climate resilience. This analysis was developed using a mixed methods approach and drawing upon substantial literature review, the researcher’s own participation in international climate policy design; elite interviews with thought leaders dealing with climate change and human rights; and regular inputs from focus groups comprised of practitioners drawn from the fields of climate change, development and human rights. This is a journal based thesis with a total of six articles submitted for evaluation, published in peer‐reviewed publications, over a five year period. Denna avhandling analyserar hur klimatfrågan och mänskliga rättigheter i samverkan formar den politiska agendan och det politiskt möjliga. Den bygger på banbrytande forskning av John Kingdon, vars publikation “Agendas, alternatives, and public policy” beskriver hur en fråga blir politiskt viktig och lyfts upp på den politiska agendan. Med utgångspunkt i Kingdons ramverk, utforskar avhandlingen hur mänskliga rättigheter bidrar till att blottlägga klimatfrågan som problem; utveckla alternativa metoder för att angripa och hantera klimatfrågan; samt skapa ett politiskt klimat nödvändigt för att på ett ambitiöst sätt kunna angripa klimatfrågan. Studien visar att klimatförändringar undergräver mänskliga rättigheterna men att arbete med mänskliga rättigheter kan vara ett effektivt verktyg för att stå emot och hantera effekterna av klimatförändringar. Analysen har genomförts med hjälp av en rad olika metoder vilka inkluderar litteraturstudier, författarens egna observationer under klimatförhandlingar; intervjuer med ledande tänkare inom klimatfrågan och mänskliga rättigheter; samt data insamlad genom fokusgrupper bestående av yrkesverksamma inom klimat, utveckling och mänskliga rättigheter. Avhandlingen är baserad på totalt sex artiklar som publicerats i fackgranskade tidskrifter under en femårsperiod.