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The liver X receptors (LXRs) are nuclear receptors that play important roles in the regulation of lipid metabolism. In this study, we demonstrate that receptor-interacting protein 140 (RIP140) is a cofactor for LXR in liver. Analysis of RIP140 null mice and hepatocytes depleted of RIP140 indicate that the cofactor is essential for the ability of LXR to activate the expression of a set of genes required for lipogenesis. Furthermore we demonstrate that RIP140 is required for the ability of LXR to repress the expression of the phosphoenolpyruvate carboxykinase gene in Fao cells and mice. Thus, we conclude that the function of RIP140 as a cofactor for LXR in liver varies according to the target genes and metabolic process, serving as a coactivator in lipogenesis but as a corepressor in gluconeogenesis.

The pollutant load in road runoff is related to traffic densities and road maintenance activities. Inurbanised areas treatment of road runoff is common and often considered necessary. The pollutantsare partitioned between the particulate and dissolved matter. However, the contaminantstend to have an affinity to the particulate material. Sedimentation, the predominant treatmentmethod for road runoff uses various types of ponds. Design tools used for stormwater treatmentsystems are based on extensive data from existing treatment systems. The variations in the empiricaldata make it difficult when attempting to evaluate precise conditions for pollutant removaland thereby minimising the land use for a treatment facility. This is a concern in highly urbanisedareas where land use often is restricted.In this work, field studies were conducted in three separate watersheds along the same motorwaywith an annual average daily traffic exceeding 120,000 vehicles. The aim was to assess treatmentconditions for the removal of contaminants from road runoff.The study of mass transport of total suspended solids used the EU Directive (1991/271/EEC)discharge requirement for urban wastewater treatment: 60 mg/l during winter and summer. Theresults showed that a capture of the total runoff volume was necessary during both seasons. Tenmetals (Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Zn), as dissolved and particulate bound, werestudied in the road runoff during a winter season and the following summer period. The dissolvedpart of Al, Cd, Co, Cr, Mn, and Ni was significantly higher in winter. The mass concentration(mg/kg) for all metals was significantly higher over the summer except for Al and Co, whichshowed a higher mass concentration during the winter. The total metal concentration showed agood correlation to total suspended solids (TSS) during winter with exception for Cd. Good correlationto TSS was also found for the summer period for Al, Cu, Fe, Mn, Ni, and Zn. A simplemodel could describe sedimentation by the initial concentration of TSS, albeit road salt (NaCl)had a significant impact on the sedimentation process during winter. Removal of dissolved metalswas studied by column experiments using water granulated blast furnace slag. The result showedgood removal for Cd, Cu, Ni, and Zn independent of NaCl concentrations. Sediment accumulation(mg sediment/mm precipitation) was relatively consistent for the studied summer seasons asopposed to winter. The sediment differed in metal mass concentrations (mg/kg) between theseasons. Concentrations of Cu and Zn were high in regard to the guidelines for sensitivity ofsediment dwelling organisms and Swedish guidelines for contaminated soils.The findings suggest that the entire runoff volume must be captured for treatment. The reductionof TSS concentration could be estimated for a specific surface load (m/h). This would alsoapply for majority of the studied metals that correlated well to the particulate material. Reactivefilter technology using water granulated blast furnace slag could be applied for treatment of runofffor the reduction of dissolved metals. However, long-term studies are necessary for its practicalimplementation. Furthermore; the work shows that on-line turbidity measurements could beused for expedient process control for treatment facilities in similar watersheds dominated byroads. The work could be used together with existing design methods and models to evaluate andoptimise road runoff treatment. Föroreningsbelastningen i vägdagvatten är beroende av trafikbelastningen och vägunderhållet. Iurbaniserade områden är behandling av dagvatten vanlig och ofta bedömd nödvändig. Föroreningarnaär lösta och partikulära, men har vanligen en affinitet till det partikulära materialet. Denförhärskande behandlingsmetoden för dagvatten är sedimentering, vanligen i dammar. Designmodellernabygger på data från olika befintliga dagvattenanläggningar. Det varierande ursprungettill det empiriska underlaget medför svårighet att precist värdera designförutsättningarna och såledesminimera behandlingsanläggningens storlek. I förtätad stadsmiljö, där tillgång på mark ärbegränsad, kan detta vara ett problem.I detta arbete har fältförsök genomförts i tre avrinningsområden vilka domineras av en motorledmed en årlig dygnstrafik större än 120,000 fordon för att utvärdera behandlingsförutsättningar förvägdagvatten.Masstransporten av suspenderat material (SS) utvärderades utifrån EU Direktivet(1991/271/EEC) och gränsvärdet för avloppsvatten på 60 mg/l under vinter och sommar. Studienvisade att hela avrinningsvolymen bör behandlas oberoende av säsong. Fördelningen mellanpartikulärt och löst material studerades för tio metaller (Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Zn)under vinter och sommar. Den lösta delen av Al, Cd, Co, Cr, Mn och Ni var signifikant högreunder vintern. Den partikulära koncentrationen (mg/kg) för samtliga metaller var högre undersommaren med undantag för Al och Co vilka förekom i högre halter under vintern. Totalhalten(μg/l) av metallerna korrelerade väl med SS under vintern med undantag för Cd. Likaledes uppvisaderesultaten en god korrelation mellan SS och Al, Cu, Fe, Mn, Ni och Zn under sommaren.Sedimenteringsegenskaperna kunde beskrivas med en enkel modell utifrån koncentration av SS,men förhöjda halter av vägsalt (NaCl) befanns påverka sedimenteringen under vintern. Reduktionav lösta metaller studerades i pilotförsök med vattenkyld granulerad masugnsslagg. God avskiljningerhölls för Cd, Cu, Ni och Zn oberoende av förhöjda halter av vägsalt. Ackumulering avsediment (mg sediment/mm nederbörd) befanns vara konstant under sommaren i motsats tillstuderade vinterperioder. Sedimentkoncentrationerna av Cu och Zn var förhöjda med avseendepå riktlinjer för känslighet hos sedimentlevande organismer samt för återanvändning av slam.Resultaten visar att hela avrinningsvolymen måste behandlas. Reduktion av SS samt huvuddelenav metallerna, vilka visade god korrelation till det partikulära materialet, kan skattas utifrån enspecifik ytbelastning (m/h). Reduktion av lösta metaller kan ske med granulerad masugnsslagg,dock bör långtidsstudier genomföras. Vidare visar studien på möjligheten att nyttja kontinuerligturbiditetsmätning för en effektiv och praktisk processkontroll i reningsanläggningar för liknandeavrinningsområden med hög trafikbelastning. Resultaten av arbetet kan användas för bedömningoch optimering av vägdagvattenbehandling tillsammans med existerande designmetoder och modeller. QC 20100709

It is known that traffic is a source of pollutants and that pollutant loads increase with elevated traffic densities. Studies executed in Stockholm, Sweden advocate that highway runoff from roads with an annual average daily traffic (AADT) exceeding 30,000 vehicles need treatment before discharge to the receiving water. It is common knowledge that sedimentation is the most expedient method for stormwater treatment. However, sedimentation units are area demanding and in highly urbanised watersheds the land use is often restricted. Studies have implied the occurrence of first flush, i.e. an initially higher pollutant load in the beginning of the runoff event, in highway watersheds. With an emphasized first flush it would be possible to treat only a part of the total runoff volume reducing the area needed for a sedimentation basin. In general two methods are used to design stormwater treatment ponds. One method is based on the reduced catchment area and pond surface and the other is based on an average runoff volume and a permanent pond volume. The methods are relaying on data from routine monitoring of various treatment systems and suggest removal efficiencies for pollutants. Applying general removal efficiencies for design it can be intricate to estimate an outlet concentration when the specific removal efficiency may be dependent on the initial concentration of the pollutant. Consequently, knowledge of the removal efficiencies dependence on initial concentration would be helpful to optimise stormwater treatment systems. This research has studied runoff from highly trafficated watersheds. The aim has been to evaluate the mass transport, stormwater quality and sedimentation behaviour and their implications for stormwater treatment. The study sites, Eugenia and Fredhäll, are located along the six-lane highway E4 through Stockholm that has an AADT load of 120,000 vehicles and a speed limit of 70 km/h. In lack of a unified definition of first flush the mass transport was studied using the EU directive 1991/271/EEC discharge demand for TSS of 60 mg/l. It was found that for the majority of the runoff events during winter the event mean concentration exceeded 60 mg/l suggesting that the complete runoff volume should be captured during winter. The dissolved concentration of metals showed significant variations between winter and summer, as did the concentration in the particulate matter (mg/kg). It was possible to correlate total metal concentration to total suspended solids with good correlation (r2 >0.90) for the majority of studied metals in winter and summer. The findings would imply that a successful treatment of the studied metal pollutants could be carried out by sedimentation. However, depending on discharge criteria, the elevated levels of dissolved matter, especially during winter, have to be considered with regards to the selection of the appropriate water treatment process. The sedimentation process could be described by a logarithmical function and initial turbidity. Good correlation (r2 >0.90) was indicated between turbidity and TSS. The sedimentation process of the studied highway runoff varied significantly (p

The yeast Mediator complex is required for transcriptional regulation both in vivo and in vitro and the identification of similar complexes from metazoans indicates that its function is conserved through evolution. Mediator subunit composition and structure is well characterized both by biochemical, genetic and biophysical methods. In contrast, little is known about the mechanisms by which Mediator operates and how the complex is regulated. The aim of my thesis was to elucidate how Mediator functions at the molecular level and to investigate functional interactions within Mediator. It is possible to recruit RNA polymerase II to a target promoter and thus to activate transcription by fusing Mediator subunits to a DNA binding domain. In order to investigate functional interactions within Mediator, we made such fusion proteins where different Mediator subunits were fused to the DNA binding domain of lexA. The expression of a reporter gene containing binding sites for lexA was subsequently measured in both a wild type strain and in strains where genes encoding specific Mediator subunits had been disrupted. We found that lexA-Med2 and lexA-Gal11 are strong activators that function independently of all Mediator subunits tested. On the other hand, lexA-Srb10 is a weak activator that depends on Srb8 and Srb11 and lexA-Med1 and lexA-Srb7 are both cryptic activators that become active in the absence of Srb8, Srb10, Srb11, or Sin4. Both lexA-Med1 and lexA-Srb7 proteins showed a stable association with the Mediator subunits Med4 and Med8 in wild type cells and in all deletion strains tested, indicating that they were functionally incorporated into the Mediator complex. We also showed that both Med4 and Med8 exist in two forms that differed in electrophoretic mobility and that these forms differed in their ability to associate with Mediator immuno-purified from the LEXA-SRB7 and LEXA-MED1 strains. Dephosphorylation assays of purified Mediator indicated that the two mobility forms of Med4 corresponded to the phosphorylated and unphosphorylated forms of the Med4 protein respectively. Some of the data presented in this study as well as previous genetic and biochemical data obtained in our lab suggested a functional link between the Med1, Med2, Srb10 and Srb11 proteins. We extended these findings by showing that the Srb10 kinase phosphorylates the Med2 protein at residue serine 208, both in vitro and in vivo. We also showed that a point mutation of the single phosphorylation site to an alanine or to an aspartic acid residue altered the gene expression of a specific set of genes. Taken together, these data indicate that posttranslational modification of Mediator subunits is a so far uncharacterized mechanism for regulation of gene expression. In order to study the function of the Srb7 subunit of Mediator, we isolated a temperature sensitive strain where the amino acids 2 to 8 of srb7 were deleted. The Mediator subunits Nut2 and Med7 were isolated as high copy suppressor of srb7-∆(2-8) and we were also able to show that Srb7 interacted with Nut2 and Med7 both in a 2-hybrid system and in co-immuno precipitation experiments using recombinantly expressed proteins. Interestingly, a deletion of amino acids 2 to 8 of Srb7 abolishes its interaction with both Med7 and Nut2 in vitro. Med4 also interacted with Srb7 in the 2-hybrid system and surprisingly, the first eight amino acids of Srb7 were shown to be sufficient for this interaction.