Molecular mechanisms of cancer predisposition in HNPCC/Lynch syndrome

Hereditary non-polyposis colorectal carcinoma (HNPCC; Lynch syndrome) is among the most common hereditary cancers in man and a model of cancers arising through deficient DNA mismatch repair (MMR). It is inherited in a dominant manner with predisposing germline mutations in the MMR genes, mainly MLH1, MSH2, MSH6 and PMS2. Both copies of the MMR gene need to be inactivated for cancer development. Since Lynch syndrome family members are born with one defective copy of one of the MMR genes in their germline, they only need to acquire a so called second hit to inactivate the MMR gene. Hence, they usually develop cancer at an early age. MMR gene inactivation leads to accumulation of mutations particularly in short repeat tracts, known as microsatellites, causing microsatellite instability (MSI). MSI is the hallmark of Lynch syndrome tumors, but is present in approximately 15% of sporadic tumors as well. There are several possible mechanisms of somatic inactivation (i.e. the second hit ) of MMR genes, for instance deletion of the wild-type copy, leading to loss of heterozygosity (LOH), methylation of promoter regions necessary for gene transcription, or mitotic recombination or gene conversion. In the Lynch syndrome tumors carrying germline mutations in the MMR gene, LOH was found to be the most frequent mechanism of somatic inactivation in the present study. We also studied MLH1/MSH2 deletion carriers and found that somatic mutations identical to the ones in the germline occurred frequently in colorectal cancers and were also present in extracolonic Lynch syndrome-associated tumors. Chromosome-specific marker analysis implied that gene conversion, rather than mitotic recombination or deletion of the respective gene locus accounted for wild-type inactivation. Lynch syndrome patients are predisposed to certain types of cancers, the most common ones being colorectal, endometrial and gastric cancer. Gastric cancer and uroepithelial tumors of bladder and ureter were observed to be true Lynch syndrome tumors with MMR deficiency as the driving force of tumorigenesis. Brain tumors and kidney carcinoma, on the other hand, were mostly MSS, implying the possibility of alternative routes of tumor development. These results present possible implications in clinical cancer surveillance. In about one-third of families suspected of Lynch syndrome, mutations in MMR genes are not found, and we therefore looked for alternative mechanisms of predisposition. According to our results, large genomic deletions, mainly in MSH2, and germline epimutations in MLH1, together explain a significant fraction of point mutation-negative families suspected of Lynch syndrome and are associated with characteristic clinical and family features. Our findings have important implications in the diagnosis and management of Lynch syndrome families.

Stabilization of Phospholipid Coatings in Capillary Electrophoresis

The development of a simple method of coating a semi-permanent phospholipid layer onto a capillary for electrochromatography use was the focus of this study. The work involved finding good coating conditions, stabilizing the phospholipid coating, and examining the effect of adding divalent cations, cetyltrimethylammonium bromide, and polyethylene glycol (PEG)-lipids on the stability of the coating. Since a further purpose was to move toward more biological membrane coatings, the capillaries were also coated with cholesterol-containing liposomes and liposomes of red blood cell ghost lipids. Liposomes were prepared by extrusion, and large unilamellar vesicles with a diameter of about 100 nm were obtained. Zwitterionic phosphatidylcholine (PC) was used as a basic component, mainly 1-palmitoyl-2-oleyl-sn-glycero-3-phosphocholine (POPC) but also eggPC and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). Different amounts of sphingomyelin, bovine brain phosphatidylserine, and cholesterol were added to the PC. The stability of the coating in 40 mM N-(2-hydroxyethyl)piperazine-N’-(2-ethanesulfonic acid) (HEPES) solution at pH 7.4 was studied by measuring the electroosmotic flow and by separating neutral steroids, basic proteins, and low-molar-mass drugs. The presence of PC in the coating solution was found to be essential to achieving a coating. The stability of the coating was improved by the addition of negative phosphatidylserine, cholesterol, divalent cations, or PEGylated lipids, and by working in the gel-state region of the phospholipid. Study of the effect on the PC coating of divalent metal ions calcium, magnesium, and zinc showed a molar ratio of 1:3 PC/Ca2+ or PC/Mg2+ to give increased rigidity to the membrane and the best coating stability. The PEGylated lipids used in the study were sterically stabilized commercial lipids with covalently attached PEG chains. The vesicle size generally decreased when PEGylated lipids of higher molar mass were present in the vesicle. The predominance of discoidal micelles over liposomes increased PEG chain length and the average size of the vesicles thus decreased. In the capillary electrophoresis (CE) measurements a highly stable electroosmotic flow was achieved with 20% PEGylated lipid in the POPC coating dispersion, the best results being obtained for disteroyl PEG (3000) conjugates. The results suggest that smaller particles (discoidal micelles) result in tighter packing and better shielding of silanol groups on the silica wall. The effect of temperature on the coating stability was investigated by using DPPC liposomes at temperatures above (45 C) and below (25 C) the main phase transition temperature. Better results were obtained with DPPC in the more rigid gel state than in the fluid state: the electroosmotic flow was heavily suppressed and the PC coating was stabilized. Also dispersions of DPPC with 0−30 mol% of cholesterol and sphingomyelin in different ratios, which more closely resemble natural membranes, resulted in stable coatings. Finally, the CE measurements revealed that a stable coating is formed when capillaries are coated with liposomes of red blood cell ghost lipids.

Radikalcyklisering vid syntes av polycykliska indoler och pyrroler

I de senaste två decennierna har radikalcyklisering, intramolekylär radikaladdition, utvecklats till en viktig syntesmetod för polycykliska indoler och pyrroler. De erhållna produktmolekylerna eller deras derivat är ofta naturliga eller syntetiska alkaloider som väckt biologiskt eller medicinskt intresse. Avhandlingen behandlar både intramolekylära radikaladditioner av pyrrolyl-, indolyl- eller indolylacylradikaler till -bindningar och intramolekylära additioner av flera olika radikaler till indolens eller pyrrolens -system. Också cykliseringar som delreaktioner i radikalkaskader behandlas. Radikalreaktioner kan släckas både oxidativt och reduktivt. För att bibehålla heteroarenens aromaticitet måste cykliseringar till pyrrol- eller indolringen släckas oxidativt. Oxidativa radikaladditioner till aromater benämns homolytiska aromatiska substitutioner. Det finns olika sätt att erhålla en reaktantradikal från en radikalprekursor. I vissa fall har radikalprekursorn en mycket labil bindning som kan brytas fotokemiskt eller med hjälp av en initiator. Till exempel kol-svavelbindningen av en O-etyl-S-alkylxanthat kan brytas på detta sätt. Ofta används dock en radikalmediator för att bilda en reaktantradikal från dess prekursor. Mediatorer är ofta föreningar, som under reaktionsförhållandena själv bildar radikaler med en stor affinitet för en prekursors specifika atom eller atomgrupp vilken abstraheras. Således fungerar mediatorn som mellanhand vid bildning av reaktantradikalen. Exempel på mediatorer av detta slag som använts vid syntes av polycykliska pyrroler och indoler är tributyltennhydrid, hexabutylditenn, tris(trimetylsilyl)silan, tributylgermaniumhydrid, dicumylperoxid, trietylboran, natriumarensulfinater (med ättiksyra) och Se-fenyl-p-toluenselenosulfonat. Också dimetylsulfoxid kan ses som mediator då den bildar metylradikaler vid Fentonreaktion i lösningsmedlet. Övergångsmetallsalter kan även bilda reaktantradikalen från prekursorn genom enelektronoxidationer eller -reduktioner. Vid syntes av polycykliska pyrroler och indoler har reaktantradikalen bildats genom enelektronoxidationer med Mn(OAc)3 eller Ag2+ (Miniscireaktion) och elektronreduktioner med ett Ni(I)-komplex eller SmI2. Avhandlingen är indelad enligt reagenset eller reagensen, som åstadkommer bildning av reaktantradikalen vid syntes av polycykliska indoler och pyrroler. Cirka hälften av avhandlingen behandlar tributyltennhydridmedierade cykliseringar då reagenset trots dess toxicitet är det överlägset mest använda. Avhandlingen diskuterar mekanismen för bildning av reaktantradikalen från prekursorn, cykliseringen och dess möjliga regioselektivitet, andra radikalreaktioner vid radikalkaskader och hur produktradikalen släcks.

Aza-Wittigringslutningar vid syntes av sammansmälta kväveheterocykler

Denna pro gradu avhandling är en litteraturstudie av intramolekylära aza-Wittigringslutningar vid syntes av sammansmälta kväveheterocykler. I arbetet behandlas material huvudsakligen från 1980 och framöver. Aza-Wittigreaktionen påminner om den analoga Wittigreaktionen. Aza-Wittigreaktioner har nästan uteslutande gjorts mellan karbonylgrupper och iminofosforaner. Reaktionsmekanismen sker enligt en tvåstegsaddition, som inleds av iminokvävets nukleofila attack till karbonylkolet och avslutas av att en zwitterjonisk betain bildar en azoxafosfetanintermediär. Intermediären sönderfaller spontant till en iminoprodukt och fosfinoxid. Reaktionen görs ofta under milda betingelser d.v.s. vid rumstemperatur och dessutom utan dyr arbetsutrustning Ett vanligt sätt att framställa iminofosforanen är från azid genom Staudingerreaktion och ofta kombineras Staudinger- och aza-Wittigreaktionerna så att iminofosforanen inte isoleras före ringslutningen. Alla andra ringslutningsreaktioner än aza-Wittig, såsom elektrocykliska ringslutningar, har uteslutits ur detta arbete. Materialet i litteraturstudien har indelats enligt vilken typ av karbonylgrupp iminofosforanen reagerar med. På så vis åskådliggörs hurudana typiska produktmolekyler som erhållits med ringslutning till en viss typ av karbonylgrupp. Det har visat sig att det är förmånligt om karbonylkolet har elektronunderskott och iminofosforanens kväve har elektronöverskott. Den ringslutande molekylens entropi och närbelägna substituenters elektroniska och steriska natur samt produktmolekylens termodynamiska fördelaktighet inverkar tillsammans på reaktionens gång. Ifall reaktionen förväntas ske långsamt är det bättre att använda sig av alkyliminofosforaner än aryliminofosforaner. Valet av lösningsmedel har nästan uteslutande lämnats oförklarat i de behandlade publikationerna men i de flesta fall har ortoxylen eller toluen varit goda lösningsmedel. Oönskad tetrazolbildning av aziden kan minimeras genom användning av opolärt lösningsmedel. Likaså kan förmånligt placerade kväveskyddsgrupper hindra intramolekylära vätebindningar. På senare tid har aza-Wittigreaktionen allt mera tillämpats vid framställning av farmakologiska produkter, vilket ökat intresset för att framställa stora mängder närbesläktade produktmolekyler. Dylika molekylbibliotek har med fördel framställts i fastfas varvid reningen av produkten underlättats märkbart. Ett nytt område inom aza-Wittigsyntetiken är asymmetriska reaktioner, vilka säkert kommer att få mera uppmärksamhet i framtiden. I denna litteraturstudie framkom det att många av de utförda synteserna på området kunde upprepas med större variation och systematik gällande reagens och reaktionsbetingelser.

Measuring and tracking quality factors in Free and Open Source Software projects

Free and Open Source Software (FOSS) has gained increased interest in the computer software industry, but assessing its quality remains a challenge. FOSS development is frequently carried out by globally distributed development teams, and all stages of development are publicly visible. Several product and process-level quality factors can be measured using the public data. This thesis presents a theoretical background for software quality and metrics and their application in a FOSS environment. Information available from FOSS projects in three information spaces are presented, and a quality model suitable for use in a FOSS context is constructed. The model includes both process and product quality metrics, and takes into account the tools and working methods commonly used in FOSS projects. A subset of the constructed quality model is applied to three FOSS projects, highlighting both theoretical and practical concerns in implementing automatic metric collection and analysis. The experiment shows that useful quality information can be extracted from the vast amount of data available. In particular, projects vary in their growth rate, complexity, modularity and team structure.

Förutsättningarna för psykiatrisk vård oberoende av patientens vilja och patientens rättsskydd vid fattandet av vårdbeslutet

I arbetet analyseras de i mentalvårdslagen (1990/1116) stadgade förutsättningarna för psykiatrisk sjukhusvård oberoende av patientens vilja gällande myndiga patienter för vilka inte har utsetts en intressebevakare före fattandet av vårdbeslutet, samt bakgrunden till och en del av de svårigheter som förknippas med denna form av vård och härtill anknutet beslutsfattande. Problemen tillspetsas framförallt i situationer av intressekonflikt mellan patientens intressen respektive samhällets intressen. Utgångspunkten ligger främst i finländsk mentalvårdslagstiftning och rättspraxis, men för att på ett ändamålsenligt sätt kunna redogöra för bakgrunden till och förutsättningarna för dylikt förvaltningsrättsligt beslutsfattande görs även relevanta kopplingar till medicinska (psykiatriska) yrkesetiska riktlinjer och mentalvårdslagstiftning inom övriga Norden. Analysen omfattar även den Europeiska människorättsdomstolens praxis i fall som gäller frihetsberövande på basis av psykiatrisk vård, närmare bestämt artikel 5 (1)(e) i den europeiska människorättskonventionen. Patientens självbestämmanderätt, frihet och integritet är utgångspunkterna för alla slags vårdförhållanden. Dessa inbegriper bl.a. frivillighet och informerat samtycke då en person söker vård. Möjligheten att förordna en person till psykiatrisk vård oberoende av dennes vilja är ett lagstadgat undantag till dessa principer. Beslutsfattandet är bundet till strikta lagstadgade förutsättningar, vilkas bedömning kräver juridisk, men framförallt medicinsk kunskap, vilket också skapar gränser och utmaningar för de yrkespersoner som arbetar med dylika ärenden. Kombinationen av psykiatri, etik och juridik är nödvändig, men inte alltid enkel att göra, vilket märks inte minst på terminologin som tillämpas i mentalvårdslagstiftningen. I arbetet analyseras en del av den terminologi som tillämpas inom finsk och övrig nordisk mentalvårdslagstiftning. Syftet är att visa att definitionerna, tolkningarna, terminologin och synsättet på psykisk ohälsa som tillämpas inom den finländska mentalvårdslagstiftningen jämförelsevis kanske inte är de mest lämpliga eller tidsenliga. En annan svår uppgift, som framförallt lagstiftaren ställ(t)s inför, är utformningen av mentalvårdslagstiftning som är tillräckligt flexibel för att tillåta beaktande av särdragen i varje enskilt ärende, men som samtidigt är strikt nog för att förhindra godtycke och missbruk. En person som lider av allvarlig psykisk ohälsa befinner sig oftast i en ytterst utsatt position, och är ofta begränsat eller inte alls kapabel att göra en korrekt bedömning av sig själv och sin situation. Således sker den starka betoningen av patientens självbestämmanderätt på både gott och ont. Det är kanske inte alltid i en psykiatrisk patients bästa att få bestämma själv, vilket också är en av frågorna som omfattas av analysen i arbetet. Eftersom det psykiatriska vårdbeslutet till karaktären är ett (skriftligt) förvaltningsbeslut, skiljer det sig samtidigt från övrigt beslutsfattande inom vårdsektorn, vilket till största delen utgör faktisk förvaltningsverksamhet. För patienten innebär vårdbeslutet också ett administrativt frihetsberövande. Ingreppet i patientens grundläggande fri- och rättigheter är särskilt stort och upplevs ofta som mycket kränkande, och därför ställs särskilt strikta krav på iakttagande av det lagstadgade beslutsförfarandet och vårdförutsättningarna samt tillgodoseendet av patientens rättsskydd. Den offentliga hälso- och sjukvården är verksamhet som är underställd den offentliga förvaltningen, vilket innebär att de allmänna förvaltningsrättsliga rättsskyddsgarantierna är tillämpliga. I och med vårdbeslutets karaktär av förvaltningsbeslut, kan ändring i beslutet sökas genom besvär hos förvaltningsdomstolen. Dessa är också några av de frågor som behandlas i arbetet.

Större frihet - bättre lokalt anpassad service? : Förvaltningsreformer och kommunernas service för utvecklingsstörda 1994-2000

In the beginning of the 1990s the legislation regarding the municipalities and the system of central government transfers were reformed in Finland. This resulted in a move from detailed governmental control to increased municipal autonomy. The purpose of this decentralization was to enable the municipalities to better adapt their administration and service supply to local needs. The aim of this study was to explore the effects of the increased municipal autonomy on the organization of services for people with intellectual disabilities. Did the increased autonomy cause the municipalities to alter their service supply and production and did the services become more adapted to local needs? The data consists of statistical information on service use and production, and also of background data such as demographics, economics and political elections on 452 municipalities in Finland from the years 1994 and 2000. The methods used are cluster analysis, discriminant analysis and factor analysis. The municipalities could be grouped in two categories: those which offered mainly one kind of residential services and others which had more varied mixes of services. The use of institutional care had decreased and municipalities which used institutional care as their primary form of service were mostly very small municipalities in 2000. The situation had changed from 1994, when institutional care was the primary service for municipalities of all sizes. Also the service production had become more differentiated and the municipalities had started using more varied ways of production. More municipalities had started producing their own services and private production had increased as well. Furthermore, the increase in local autonomy had opened up possibilities for local politics to influence both the service selection and methods of production. The most significant motive for changes in the service structure was high unemployment and an increasing share of elderly people in the population, particularly in sparsely populated areas. Municipalities with a low level of resources had made more changes in their service organization while those with more resources had been able to carry on as before. Key words: service structure, service for people with intellectual disabilities, municipalities, contingency theory, New Public Management

Redox-linked proton transfer by cytochrome c oxidase

The respiratory chain is found in the inner mitochondrial membrane of higher organisms and in the plasma membrane of many bacteria. It consists of several membrane-spanning enzymes, which conserve the energy that is liberated from the degradation of food molecules as an electrochemical proton gradient across the membrane. The proton gradient can later be utilized by the cell for different energy requiring processes, e.g. ATP production, cellular motion or active transport of ions. The difference in proton concentration between the two sides of the membrane is a result of the translocation of protons by the enzymes of the respiratory chain, from the negatively charged (N-side) to the positively charged side (P-side) of the lipid bilayer, against the proton concentration gradient. The endergonic proton transfer is driven by the flow of electrons through the enzymes of the respiratory chain, from low redox-potential electron donors to acceptors of higher potential, and ultimately to oxygen. Cytochrome c oxidase is the last enzyme in the respiratory chain and catalyzes the reduction of dioxygen to water. The redox reaction is coupled to proton transport across the membrane by a yet unresolved mechanism. Cytochrome c oxidase has two proton-conducting pathways through which protons are taken up to the interior part of the enzyme from the N-side of the membrane. The K-pathway transfers merely substrate protons, which are consumed in the process of water formation at the catalytic site. The D-pathway transfers both substrate protons and protons that are pumped to the P-side of the membrane. This thesis focuses on the role of two conserved amino acids in proton translocation by cytochrome c oxidase, glutamate 278 and tryptophan 164. Glu278 is located at the end of the D-pathway and is thought to constitute the branching point for substrate and pumped protons. In this work, it was shown that although Glu278 has an important role in the proton transfer mechanism, its presence is not an obligatory requirement. Alternative structural solutions in the area around Glu278, much like the ones present in some distantly related heme-copper oxidases, could in the absence of Glu278 support the formation of a long hydrogen-bonded water chain through which proton transfer from the D-pathway to the catalytic site is possible. The other studied amino acid, Trp164, is hydrogen bonded to the ∆-propionate of heme a3 of the catalytic site. Mutation of this amino acid showed that it may be involved in regulation of proton access to a proton acceptor, a pump site, from which the proton later is expelled to the P-side of the membrane. The ion pair that is formed by the ∆-propionate of heme a3 and arginine 473 is likely to form a gate-like structure, which regulates proton mobility to the P-side of the membrane. The same gate may also be part of an exit path through which water molecules produced at the catalytically active site are removed towards the external side of the membrane. Time-resolved optical and electrometrical experiments with the Trp164 to phenylalanine mutant revealed a so far undetected step in the proton pumping mechanism. During the A to PR transition of the catalytic cycle, a proton is transferred from Glu278 to the pump site, located somewhere in the vicinity of the ∆-propionate of heme a3. A mechanism for proton pumping by cytochrome c oxidase is proposed on the basis of the presented results and the mechanism is discussed in relation to some relevant experimental data. A common proton pumping mechanism for all members of the heme-copper oxidase family is moreover considered.

Population dynamics of blue mussels in a variable environment at the edge of their range

Spatial and temporal variation in the abundance of species can often be ascribed to spatial and temporal variation in the surrounding environment. Knowledge of how biotic and abiotic factors operate over different spatial and temporal scales in determining distribution, abundance, and structure of populations lies at the heart of ecology. The major part of the current ecological theory stems from studies carried out in central parts of the distributional range of species, whereas knowledge of how marginal populations function is inadequate. Understanding how marginal populations, living at the edge of their range, function is however in a key position to advance ecology and evolutionary biology as scientific disciplines. My thesis focuses on the factors affecting dynamics of marginal populations of blue mussels (Mytilus edulis) living close to their tolerance limits with regard to salinity. The thesis aims to highlight the dynamics at the edge of the range and contrast these with dynamics in more central parts of the range in order to understand the potential interplay between the central and the marginal part in the focal system. The objectives of the thesis are approached by studies on: (1) factors affecting regional patterns of the species, (2) long-term temporal dynamics of the focal species spaced along a regional salinity gradient, (3) selective predation by increasing populations of roach (Rutilus rutilus) when feeding on their main food item, the blue mussel, (4) the primary and secondary effects of local wave exposure gradients and (5) the role of small-scale habitat heterogeneity as determinants of large-scale pattern. The thesis shows that populations of blue mussels are largely determined by large scale changes in sea water salinity, affecting mainly recruitment success and longevity of local populations. In opposite to the traditional view, the thesis strongly indicate that vertebrate predators strongly affect abundance and size structure of blue mussel populations, and that the role of these predators increases towards the margin where populations are increasingly top-down controlled. The thesis also indicates that the positive role of biogenic habitat modifiers increases towards the marginal areas, where populations of blue mussels are largely recruitment limited. Finally, the thesis shows that local blue mussel populations are strongly dependent on high water turbulence, and therefore, dense populations are constrained to offshore habitats. Finally, the thesis suggests that ongoing sedimentation of rocky shores is detrimental for the species, affecting recruitment success and post-recruit survival, pushing stable mussel beds towards offshore areas. Ongoing large scale changes in the Baltic Sea, especially dilution processes with attendant effects, are predicted to substantially contract the distributional range of the mussel, but also affect more central populations. The thesis shows that in order to understand the functioning of marginal populations, research should (1) strive for multi-scale approaches in order to link ecosystem patterns with ecosystem processes, and (2) challenge the prevailing tenets that origin from research carried out in central areas that may not be valid at the edge.

Spatial ecology of a specialist insect herbivore : the leaf-mining moth Tischeria ekebladella on the pedunculate oak Quercus robur

Herbivorous insects comprise a major part of terrestrial biodiversity, and their interactions with their host plants and natural enemies are of vast ecological importance. A large body of research demonstrates that the ecology and evolution of these insects may be affected by trophic interactions, by abiotic influences, and by intraspecific processes, but so far research on these individual aspects has rarely been combined. This thesis uses the leaf-mining moth Tischeria ekebladella and the pedunculate oak (Quercus robur) as a case study to assess how spatial variation in trophic interactions and the physical distribution of host trees jointly affect the distribution, dynamics and evolution of a host-specific herbivore. With respect to habitat quality, Tischeria ekebladella experiences abundant variation at several spatial scales. Most of this variation occurs at small scales notably among leaves and shoots within individual trees. While hypothetically this could cause moths to evolve an ability to select leaves and shoots of high quality, I did not find any coupling between female preference and offspring performance. Based on my studies on temporal variation in resource quality I therefore propose that unpredictable temporal changes in the relative rankings of individual resource units may render it difficult for females to predict the fate of their developing offspring. With respect to intraspecific processes, my results suggest that limited moth dispersal in relation to the spatial distribution of oak trees plays a key role in determining the regional distribution of Tischeria ekebladella. The distribution of the moth is aggregated at the landscape level, where local leaf miner populations are less likely to be present where oaks are scarce. A modelling exercise based on empirical dispersal estimates revealed that the moth population on Wattkast an island in south-western Finland is spatially structured overall, but that the relative importance of local and regional processes on tree-specific moth dynamics varies drastically across the landscape. To conclude, my work in the oak-Tischeria ekebladella system demonstrates that the local abundance and regional distribution of a herbivore may be more strongly influenced by the spatial location of host trees than by their relative quality. Hence, it reveals the importance of considering spatial context in the study of herbivorous insects, and forms a bridge between the classical fields of plant-insect interactions and spatial ecology.

Lymphatic vascular maturation : Roles of FOXC2, NFATc1 and liprin ß1

The blood vascular system is a closed circulatory system, responsible for delivering oxygen and nutrients to the tissues. In contrast, the lymphatic vascular system is a blind-ended transport system that collects the extravasated tissue fluid from the capillary beds, and transports it back to the blood circulation. Failure in collecting or transporting the lymph, due to defects in the lymphatic vasculature, leads to accumulation of extra fluid in the tissues, and consequently to tissue swelling lymphedema. The two vascular systems function in concert. They are structurally related, but their development is regulated by separate, however overlapping, molecular mechanisms. During embryonic development, blood vessels are formed by vasculogenesis and angiogenesis, processes largely mediated by members of the vascular endothelial growth factor (VEGF) family and their tyrosine kinase receptors. The lymphatic vessels are formed after the cardiovascular system is already functional. This process, called lymphangiogenesis, is controlled by distinct members of the VEGF family, together with the transcription factors Prox1 and Sox18. After the primary formation of the vessels, the vasculature needs to mature and remodel into a functional network of hierarchically organized vessels: the blood vasculature into arteries, capillaries and veins; and the lymphatic vasculature into lymphatic capillaries, responsible for the uptake of the extravasated fluid from the tissues, and collecting vessels, responsible for the transport of the lymph back to the blood circulation. A major event in the maturation of the lymphatic vasculature is the formation of collecting lymphatic vessels. These vessels are characterized by the presence of intraluminal valves, preventing backflow of the lymph, and a sparse coverage of smooth muscle cells, which help in pumping the lymph forward. In our study, we have characterized the molecular and morphological events leading to formation of collecting lymphatic vessels. We found that this process is regulated cooperatively by the transcription factors Foxc2 and NFATc1. Mice lacking either Foxc2 or active NFATc1 fail to remodel the primary lymphatic plexus into functional lymphatic capillaries and collecting vessels. The resulting vessels lack valves, display abnormal expression of lymphatic molecules, and are hyperplastic. Moreover, the lymphatic capillaries show aberrant sprouting, and are abnormally covered with smooth muscle cells. In humans, mutations in FOXC2 lead to Lymphedema-Distichiasis (LD), a disabling disease characterized by swelling of the limbs due to insufficient lymphatic function. Our results from Foxc2 mutant mice and LD patients indicate that the underlying cause for lymphatic failure in LD is agenesis of collecting lymphatic valves and aberrant recruitment of periendothelial cells and basal lamina components to lymphatic capillaries. Furthermore, we show that liprin β1, a poorly characterized member of the liprin family of cytoplasmic proteins, is highly expressed in lymphatic endothelial cells in vivo, and is required for lymphatic vessel integrity. These data highlight the important role of FOXC2, NFATc1 and liprin β1 in the regulation of lymphatic development, specifically in the maturation and formation of the collecting lymphatic vessels. As damage to collecting vessels is a major cause of lymphatic dysfunction in humans, our results also suggest that FOXC2 and NFATc1 are potential targets for therapeutic intervention.

Macroseismology as a Component of Seismicity Assessments in an Intraplate Region: Studies of Northern Europe with Emphasis on Finland

This work focuses on the role of macroseismology in the assessment of seismicity and probabilistic seismic hazard in Northern Europe. The main type of data under consideration is a set of macroseismic observations available for a given earthquake. The macroseismic questionnaires used to collect earthquake observations from local residents since the late 1800s constitute a special part of the seismological heritage in the region. Information of the earthquakes felt on the coasts of the Gulf of Bothnia between 31 March and 2 April 1883 and on 28 July 1888 was retrieved from the contemporary Finnish and Swedish newspapers, while the earthquake of 4 November 1898 GMT is an example of an early systematic macroseismic survey in the region. A data set of more than 1200 macroseismic questionnaires is available for the earthquake in Central Finland on 16 November 1931. Basic macroseismic investigations including preparation of new intensity data point (IDP) maps were conducted for these earthquakes. Previously disregarded usable observations were found in the press. The improved collection of IDPs of the 1888 earthquake shows that this event was a rare occurrence in the area. In contrast to earlier notions it was felt on both sides of the Gulf of Bothnia. The data on the earthquake of 4 November 1898 GMT were augmented with historical background information discovered in various archives and libraries. This earthquake was of some concern to the authorities, because extra fire inspections were conducted in three towns at least, i.e. Tornio, Haparanda and Piteå, located in the centre of the area of perceptibility. This event posed the indirect hazard of fire, although its magnitude around 4.6 was minor on the global scale. The distribution of slightly damaging intensities was larger than previously outlined. This may have resulted from the amplification of the ground shaking in the soft soil of the coast and river valleys where most of the population was found. The large data set of the 1931 earthquake provided an opportunity to apply statistical methods and assess methodologies that can be used when dealing with macroseismic intensity. It was evaluated using correspondence analysis. Different approaches such as gridding were tested to estimate the macroseismic field from the intensity values distributed irregularly in space. In general, the characteristics of intensity warrant careful consideration. A more pervasive perception of intensity as an ordinal quantity affected by uncertainties is advocated. A parametric earthquake catalogue comprising entries from both the macroseismic and instrumental era was used for probabilistic seismic hazard assessment. The parametric-historic methodology was applied to estimate seismic hazard at a given site in Finland and to prepare a seismic hazard map for Northern Europe. The interpretation of these results is an important issue, because the recurrence times of damaging earthquakes may well exceed thousands of years in an intraplate setting such as Northern Europe. This application may therefore be seen as an example of short-term hazard assessment.

Quantum Chemical Studies on Tropospheric Nucleation Mechanisms Involving Sulfuric Acid

Nucleation is the first step of the process by which gas molecules in the atmosphere condense to form liquid or solid particles. Despite the importance of atmospheric new-particle formation for both climate and health-related issues, little information exists on its precise molecular-level mechanisms. In this thesis, potential nucleation mechanisms involving sulfuric acid together with either water and ammonia or reactive biogenic molecules are studied using quantum chemical methods. Quantum chemistry calculations are based on the numerical solution of Schrödinger's equation for a system of atoms and electrons subject to various sets of approximations, the precise details of which give rise to a large number of model chemistries. A comparison of several different model chemistries indicates that the computational method must be chosen with care if accurate results for sulfuric acid - water - ammonia clusters are desired. Specifically, binding energies are incorrectly predicted by some popular density functionals, and vibrational anharmonicity must be accounted for if quantitatively reliable formation free energies are desired. The calculations reported in this thesis show that a combination of different high-level energy corrections and advanced thermochemical analysis can quantitatively replicate experimental results concerning the hydration of sulfuric acid. The role of ammonia in sulfuric acid - water nucleation was revealed by a series of calculations on molecular clusters of increasing size with respect to all three co-ordinates; sulfuric acid, water and ammonia. As indicated by experimental measurements, ammonia significantly assists the growth of clusters in the sulfuric acid - co-ordinate. The calculations presented in this thesis predict that in atmospheric conditions, this effect becomes important as the number of acid molecules increases from two to three. On the other hand, small molecular clusters are unlikely to contain more than one ammonia molecule per sulfuric acid. This implies that the average NH3:H2SO4 mole ratio of small molecular clusters in atmospheric conditions is likely to be between 1:3 and 1:1. Calculations on charged clusters confirm the experimental result that the HSO4- ion is much more strongly hydrated than neutral sulfuric acid. Preliminary calculations on HSO4- NH3 clusters indicate that ammonia is likely to play at most a minor role in ion-induced nucleation in the sulfuric acid - water system. Calculations of thermodynamic and kinetic parameters for the reaction of stabilized Criegee Intermediates with sulfuric acid demonstrate that quantum chemistry is a powerful tool for investigating chemically complicated nucleation mechanisms. The calculations indicate that if the biogenic Criegee Intermediates have sufficiently long lifetimes in atmospheric conditions, the studied reaction may be an important source of nucleation precursors.

Pengarna eller livet? : En kvalitativ och longitudinell studie om långtidsarbetslösa unga i ett aktörsperspektiv

Your money or your life? A qualitative follow-up study of the young unemployed from an actor perspective is a qualitative and longitudinal study following 36 unemployed young people in Helsinki over a span of ten years. The purpose of the study is to shed light on how a few young people view employment/unemployment and their lives and future, how they as unemployed perceive their encounters with society, and how society supports them. Four so-called key informants were followed at a finer level of empirical detail. They were chosen for the thematic interviews because of their different personalities, starting points and preferences. Although some differences were expected, what the results show is quite striking. The individual stories raise a number of questions about differences between young people, about society s view of the young unemployed, and about the principles behind the so-called activation policy and how society s support is distributed. The key informants descriptions underline that the group young unemployed does not consist of individuals who are alike but that life is complex, that paid work and unemployment can be perceived very differently, and that background and unofficial support can have consequences for self-perception and for ways of looking at the future, vocational choices, paid work and activation policy. Margaret S. Archer s theory of Morphogenesis and Barbara Cruikshank s theory of constructing democracies compose the study s theoretical framework. The key informants stories give a picture of a formal support system that, even though it puts part of the responsibility for unemployment on the individuals themselves, in the name of fairness and equality, treats them in an impersonal way, not giving their personal situation and wishes much weight. As a consequence, those who share the dominant values of society do well, while others who do not are faced with difficulties. The bigger the gap between society s and the individual s values, the bigger the risk to be met by little understanding and by penalties. And vice versa: Those who initially have the right values and know how to deal with authorities get heard and their opinions get accepted. The informants ask for a more personal encounter, which could improve both the atmosphere and the clients experiences of being heard. Still the risk of having a more individualistic system should be addressed, as a new system might generate new winners, but just as well give new losers. Finally, we have to ask if the so-called activation policy is looking for answers primarily to a macro-level problem on the micro-level. If it does not produce more jobs, its support for the unemployed will be insignificant. It is not enough to think about what to do at the grassroots level to make the system more functional and support job-seeking. If the current rate of unemployment endures, the quality of life of the unemployed should be addressed. A first step could be taken by placing less guilt on the unemployed. Instead of talking about activating the unemployed, discussion should be targeted at removing structural impediments to employment. If we want to have less polarisation between the those with paid work and those without, who often struggle with low incomes, we need to include the macro-level in the discussion. What does high unemployment mean in a work-based society, where the individual s self-perception and important social forms of support are linked to labour income? And what can be done at the macro-level to change this undesirable condition at the micro-level? Keywords: Unemployment, Youth, Public interventions, Activation policy, Individual actors, Qualitative, Longitudinal, Holistic, Helsinki, Finland