I ett mångmiljardprojekt i Stockholms innerstad, Sergelhuset, har ett nytt sätt att arbeta med arbetsplatsens omkostnader (APO) vuxit fram. Istället för att byggentreprenören tar hand om allt temporärt på arbetsplatsen har beställarna valt att upphandla det i en separat, så kallad APO-entreprenad. Det innebär ansvar för bland annat projektorganisation, logistik, säkerhet och tillfälliga etableringar. Denna rapport är en undersökning av hur byggbranschen arbetar medarbetsplatsomkostnader (APO) i produktion och huruvida Ramirents koncept Den Temporära Fabriken kan hjälpa branschens behov av ökad samordning och effektivisering i byggproduktion. I rapportens inledande del framgår att behoven är mycket stora inom APO. Det är känt att området behöver ständig uppföljning och kontroll, men det finns fortfarande stora brister i genomförandet. Vidare redovisas resultat av intervjuer med projektchefer i ett flertal av Sveriges största byggbolag, för att se om det finns ett behov av en samordningstjänst inom APO. Även ledande personer inom byggbranschen har gett utlåtande om detta nya sätt att arbeta. Slutsatsen är att om en effektivisering av byggarbetsplatsen ska kunna ta fart på allvar behöver beställare, entreprenörer och APO-leverantörer samarbeta, gemensamt dela på risker och genomföra innovativa lösningar. Branschen behöver sluta arbeta med en slags stuprörsmentalitet och se att samverkan, långsiktigt förtroende- och värdeskapande är vägen framåt. / In a multibillion project called Sergelhuset in the city center of Stockholm, a new way of handling general overhead costs and general tasks in building projects (APO), was developed. Instead of the construction companies doing it as usual, the purchaser chose to buy the service from a third party. It would amount to project organization, logistics, safety and temporary solutions. The report investigates how the construction business handles general costs and whether Ramirent’s new concept The Temporary Factory can help increase efficiency on the construction site. It also contains information that shows great needs in this area and the needs of continuous follow up and control. The report includes interviews with prominent project managers and other field experts. To reach a higher level of efficiency the purchaser, construction companies and APO-provider need to cooperate, share risks and aim for high value creation in new innovative projects.

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Le but de ma thèse est de conduire et d’utiliser les premières observations atmosphériquesde surface du CO2, de l’O2, du CH4 et de la composition isotopique de la vapeur d’eau et desprécipitations dans la région peu instrumentée du sud du Groenland. Quelles informations cesobservations locales peuvent-elles nous apporter à la fois concernant les processus atmosphériqueslocaux et concernant le transport atmosphérique à grande échelle ? Peut-on en extrairedes informations sur les échanges entre la surface et l’atmosphère, qui constituent une part crucialedes cycles biogéochimiques du carbone et de l’eau ? L’interprétation paléoclimatique desenregistrements de carottes de glace au Groenland peut-elle bénéficier de ces observations ?Après une validation des observations menées à Ivittuut, sur la côte sud-ouest du Groenland,en regard de normes internationales de qualité, confirmant leur représentativité des variationsatmosphériques, j’ai commencé par interpréter ces observations en analysant leur variabilité àdifférentes échelles de temps et découlant de processus spécifiques. Ceci m’a permis de montrerque le site de mesure était très peu influencé par des phénomènes locaux : peu de sources localesde gaz à effet de serre, et de processus locaux agissant sur la composition isotopique de la vapeurd’eau. Cette caractéristique facilite l’attribution des variations observées à des changements detransport atmosphérique de grande échelle en lien avec des sources distantes.Pour comprendre les variations observées, je les ai mises en relation avec des observationsissues d’autres sites ainsi qu’avec des sorties de modèles atmosphériques de grande échelle. D’unepart, j’ai constaté que de nombreuses variations de la composition atmosphérique à l’échelle synoptiquepouvaient être expliquées par des modifications du transport atmosphérique de grandeéchelle. J’ai aussi pu identifier la représentativité spatiale de nos observations, et contribuerà l’évaluation de modèles atmosphériques de grande échelle. Enfin, un cas d’étude durant lavague de chaleur ayant affecté la totalité du Groenland durant l’été 2012 m’a permis de suivrele transport atmosphérique de masses d’air et d’observer en plusieurs points ses impacts sur lacomposition isotopique de la vapeur d’eau, notamment sur la conservation du signal d’excès endeutérium, qui n’avait jamais été observée auparavant.Enfin, la combinaison des observations et de la modélisation atmosphérique m’a permis demettre en évidence les contributions de l’observation atmosphérique en terme de documentationdes échanges entre la surface et l’atmosphère : par une attribution des valeurs observées auxsources potentielles identifiées par modélisation des masses d’air affectant la station pour le CH4et la composition isotopique de la vapeur d’eau, et par la comparaison des observations à dessimulations de transport direct de sources connues issues d’inventaires pour le CH4 ou modéliséespour le CO2 et l’O2 océanique. / South Greenland is a key region placed under Arctic and Northern Atlantic influences, whichis poorly documented in terms of atmospheric monitoring. The aim of my thesis is to conductand use the first regional atmospheric observations of CO2, O2, CH4 and isotopic compositionof water vapour and precipitation performed in Ivittuut, a coastal site in south-west Greenland.Which information can we infer from these atmospheric observations, in terms of localatmospheric processes and large scale atmospheric transport variations? Can we use these observationsto document the surface-air exchanges of these compounds, which represent an importantpart of the water and carbon biogeochemical cycles?After validating the observations regarding international precision recommandations, I havebeen able to analyse the variabilities at different time scales, resulting from different specificprocesses. This highlighted the fact the atmospheric composition of our site is weakly influencedby local processes, either for local greenhouse gases sources or small scale atmosphericvariations affecting water vapour isotopic composition. This facilitates the interpretation of ourobservations in terms of large scale atmospheric transport signals.To better understand the observed variations, I related our data series with other observationsoriginating from different sites, and with outputs from different atmospheric models. Ihave first witnessed the links between variations of atmospheric composition and large synopticscale atmospheric transport changes. Then, I have identified the spatial representativity of ourobservations and contributed to the evaluation of atmospheric models. The case study of astrong heat wave covering Greenland during summer 2012 allowed me to study the transport ofwater vapour within an air mass between different stations. It allowed me to provide the firstexperimental observation of deuterium excess conservation during atmospheric transport.Finally, combining observations and atmospheric simulations, I have documented surfaceairexchanges of the observed compounds: first by the attribution of observed values of CH4and water vapour isotopic composition to their simulated potential origins, secondly by thecomparison of observed variations with the direct simulation of atmospheric transport of eitherCH4 sources estimated from inventories or simulated CO2 and O2 air-sea fluxes from an oceanmodel.

Die Identifizierung von Apoptose-modifizierten Proteinen erfolgte durch Proteomanalyse per 2D-Gelelektrophorese und Massenspektrometrie. Zunächst wurde eine 2DE-Datenbank errichtet, die im Internet zugänglich ist. Die Identifikation von Proteinen durch Peptidmassenfingerabdruck konnte durch Verwendung einer zweiten Matrix verbessert werden. Die Analyse ausgelassener tryptischer Spaltstellen, N-terminaler Pyroglutamatbildung und Oxidation von Tryptophan konnte die Identifikation von Proteinen mittels Peptidmassenfingerabdruck verbessern. Die Apoptose wurde über den Fas-Rezeptor-Signalweg oder durch DNA-Schädigung mittels Cis-Platin eingeleitet und das Totallysat oder die Kompartimente von Jurkat T-Zellen der Proteomanalyse unterworfen. Große Übereinstimmungen der beiden Prozesse wurden festgestellt. 95 Apoptose-modifizierte Proteine wurden identifiziert, wovon 78 Proteine bisher unbekannt für den Apoptoseprozess waren. Auffällig war, dass 40 % der Proteine RNA-Bindungsmotive enthielten und das 21 Onkoprotein oder Onkoprotein-interagierende Proteine identifiziert wurden. Für 39 Proteine konnten bisher proteolytische Spaltungen vorausgesagt werden. Eine Fülle von Informationen wurde über putative Translokationen der Proteine erhalten. Für das Protein p54nrb wurden drei Caspase-3 Spaltstellen durch die Einführung von Mutationen und die Abhängigkeit der Caspase-3 Spaltung von RNA bewiesen. Mitochondriale ribosomale Proteine von Mensch, Maus und Ratte wurden durch Abgleichung von EST-Datenbanken mit partiellen Aminosäuresequenzen aus dem Rind bestimmt. Die Konservierung der Sequenzen der Säugetierproteine der mitochondrialen ribosomalen Proteinen war geringer als von den bekannten cytosolischen ribosomalen Proteinen. Weiterhin wurden unterschiedliche Ergebnisse bzgl. der mitochondrialen Signalsequenzen der Proteine gefunden. RNA-Protein-Wechselwirkungen im Ribosom wurden nach Quervernetzung auf einzelne Aminosäuen bzw. Nukleotide bestimmt. Die Daten wurden zur Verbesserung von ribosomalen Modellen verwendet. Die mittlerweile erhaltenen Kristallstrukturen des Ribosoms zeigten, dass die Ergebnisse der Quervernetzungsexperimente mit den tatsächlichen RNA-Protein-Wechselwirkungen weitgehend übereinstimmen. Die Affinität von verschiedenen Komponenten zu einem Zielmolekül zur Bildung von nicht-kovalenten RNA-Peptid-Wechselwirkungen wurde mit Hilfe von MALDI-MS ermittelt. Die Interaktionen sind stark abhängig von der Anzahl der Arginine. / Apoptosis-modified proteins were identified by proteome analysis via 2D gel electrophoresis and mass spectrometry. First, a internet-accessible 2DE database was rendered. The identification of the proteins by peptide mass fingerprinting was improved using a second matrix. The analysis of missed tryptic cleavage sites, the formation of N-terminal pyroglutamine and oxidation of tryptophan could improve the identification of proteins by peptide mass fingerprinting. Apoptosis was induced via the Fas-receptor signaling pathway or by means of DNA damage by cis-platin. The total lysate and the compartments of Jurkat T cells were analyzed by proteome analysis. High similarities between both processes were observed. 95 apoptosis-modified proteins were identified, 78 of these were until now unknown to be involved in apoptosis. Noticeable, 40% of the proteins include a RNA-binding motif and 21 oncogene or oncogene-interacting proteins were identified. A proteolytic cleavage could be predicted for 39 proteins. Some information was received about the putative translocation of the proteins. Three caspase-3 cleavage sites were shown for the protein p54nrb with the incorporation of mutations. Furthermore, the caspase-3 cleavage was dependent on the occurrence of RNA. Mitochondrial ribosomal proteins of human, mouse and rat were determined by screening of EST-databases with partial amino acid sequences from bovine. The conservation of sequences of mammalian proteins of the mitochondrial ribosomal proteins was less than for known cytosolic ribosomal proteins. Furthermore, different results were obtained considering mitochondrial signal sequences. RNA-protein interaction within the ribosome were determined on single amino acids and nucleotides, respectively, after cross-linking. These data were used to improve models of the ribosome. The in the meantime obtained 3D-structures of the ribosome showed high consistency with the revealed RNA-protein interaction sites after cross-linking. The affinity of different components to a target molecule to form RNA-peptide interactions was determined by MALDI-MS. The interactions were strongly dependent on the number of arginines.

<p>Proteins are among the most abundant biological macromolecules. The cellular machinery is coupled to exact structural shape and properties of the more than 100 000 different proteins. Still, proteins can sometimes completely change their character and as a result trigger neuro degenerative disease. Exactly what happens is yet poorly understood but misfolding and aggregation leading to toxic gain of function is probable causes, i.e. the protein adopts new noxious properties. In 1993 the protein superoxide dismutase (SOD) was found to be associated with the neuro degenerative disease ALS. Up to date more than 100 mutations in SOD have been associated with ALS. However, the mutations are scattered all over the structure and no common denominator for the disease mechanism has been found. </p><p>This work has been focused on the molecular mechanism of the toxic gain-of - function of mutant SOD from the perspective of protein folding and structural stability. To facilitate the studies of SOD and its ALS associated mutations, an expression system resulting in increased copper content was developed. Coexpression with the copper chaperone for superoxide dismutase (yCCS) leads to increased expression levels, especially for the destabilised ALS mutants. Through thermodynamic studies, I show that with the exception of the most disruptive mutations the holo protein is only marginally destabilised, whereas all mutations show a pronounced destabilisation on the apo protein. Kinetic studies suggest further that the dimeric apoSOD folds via a three-state process where the dimerisation proceeds via a marginally stable monomer. The apoSOD monomer folds by a two-state process. The disulphide bond is not critical for the folding of the apoSOD monomer although it contributes significantly to its stability. Interestingly, in the absence of metals, reduction of the disulphide bond prevents the formation of the dimer. A mutation can affect the protein stability in various ways: either from destabilisation of the monomer (case 1), weakening of the dimer interface (case 2) or, in the worst case, from a combination of both (case 1+2). Thus, therapeutic strategies to prevent the noxious effects of mutant SOD must include both mechanisms. An important finding in this study is that we can see a correlation between the stability for each mutation and the mean survival time. This could be an opening in the development of therapeutic substances that counteract the defect in SOD upon mutation.</p>

Proteins are among the most abundant biological macromolecules. The cellular machinery is coupled to exact structural shape and properties of the more than 100 000 different proteins. Still, proteins can sometimes completely change their character and as a result trigger neuro degenerative disease. Exactly what happens is yet poorly understood but misfolding and aggregation leading to toxic gain of function is probable causes, i.e. the protein adopts new noxious properties. In 1993 the protein superoxide dismutase (SOD) was found to be associated with the neuro degenerative disease ALS. Up to date more than 100 mutations in SOD have been associated with ALS. However, the mutations are scattered all over the structure and no common denominator for the disease mechanism has been found. This work has been focused on the molecular mechanism of the toxic gain-of - function of mutant SOD from the perspective of protein folding and structural stability. To facilitate the studies of SOD and its ALS associated mutations, an expression system resulting in increased copper content was developed. Coexpression with the copper chaperone for superoxide dismutase (yCCS) leads to increased expression levels, especially for the destabilised ALS mutants. Through thermodynamic studies, I show that with the exception of the most disruptive mutations the holo protein is only marginally destabilised, whereas all mutations show a pronounced destabilisation on the apo protein. Kinetic studies suggest further that the dimeric apoSOD folds via a three-state process where the dimerisation proceeds via a marginally stable monomer. The apoSOD monomer folds by a two-state process. The disulphide bond is not critical for the folding of the apoSOD monomer although it contributes significantly to its stability. Interestingly, in the absence of metals, reduction of the disulphide bond prevents the formation of the dimer. A mutation can affect the protein stability in various ways: either from destabilisation of the monomer (case 1), weakening of the dimer interface (case 2) or, in the worst case, from a combination of both (case 1+2). Thus, therapeutic strategies to prevent the noxious effects of mutant SOD must include both mechanisms. An important finding in this study is that we can see a correlation between the stability for each mutation and the mean survival time. This could be an opening in the development of therapeutic substances that counteract the defect in SOD upon mutation.

Les études épidémiologiques ont montré qu'une consommation régulière en tomate et ses produits dérivés de tomate permet de lutter contre diverses pathologies dégénératives associées notamment au stress oxydant (maladies cardiovasculaires, cancers etc..). Les effets bénéfiques pourraient être dus au lycopène pigment rouge de la tomate et/ou à ses métabolites qui interviendraient dans ce processus soit de part leurs capacités antioxydantes, soit au travers de la régulation de l'expression de gènes. Dans ce contexte, quatre familles de molécules dérivées du lycopène, pouvant être des métabolites potentiels, ont été ciblées pour la synthèse organique : les apo-11-lycopénoïdes, les apo- 14'-lycopénoïdes, les apo-12'-lycopénoïdes et les apo-10'-lycopénoïdes. Chacune des familles a été synthétisée, via des réactions de couplages tels que Wittig et Horner-Wadsworth-Emmons, avec quatre fonctions chimiques terminales : ester, acide carboxylique, alcool et aldéhyde. Par la suite deux types de propriétés physico-chimiques des composés synthétisés ont été étudiés : mesure du pouvoir antioxydant dans des conditions expérimentales mimant un stress oxydant dans le compartiment gastro-intestinal (inhibition de la peroxydation lipidique initiée par la metmyoglobine en milieu micellaire) et une étude d'interaction avec l'albumine de sérum humain, protéine impliquée dans le transport des acides gras dans le plasma.

Certaines protéines, appelées protéines amphitropiques, sont solubles en milieux aqueux et capables de se lier aux membranes pour leurs activités biologiques. Ces protéines apparaissent comme d'excellents modèles pour la compréhension des principes généraux de stabilité et de repliement des protéines membranaires intrinsèques. L'α-lactalbumine et l'apo-myoglobine ont été choisies comme modèles de protéines amphitropiques. Nous proposons une étude exhaustive des facteurs qui influencent la liaison aux membranes de ces deux protéines telles que le pH, la présence de cofacteurs, la force ionique, la charge et la courbure des membranes. Une description macroscopique des mécanismes de liaison a été entreprise par des expériences de centrifugation et de fluorescence. Enfin, la structure et la stabilité des états liés aux membranes ont été sondées par des expériences d'échanges hydrogène/deutérium (H/D).<br />L'état conformationnel de l'α-lactalbumine liée aux membranes s'est révélé très dépendant de la courbure membranaire. Nous avons identifié deux hélices amphiphiles qui sont directement impliquées dans les interactions α-lactalbumine-lipides. D'autre part, nos expériences d'échanges H/D permettent une description thermodynamique de la liaison à la membrane. Dans le cas de l'apo-myoglobine, nos résultats suggèrent un mécanisme commun d'insertion membranaire pour les protéines qui possèdent une topologie de type globine. Par ailleurs, la liaison de l'apo-myoglobine aux membranes ne semble pas nécessaire pour la fonction biologique de cette protéine. Une description des structures membranaires de l'apo-myoglobine a été entreprise par des expériences d'échanges H/D suivies par spectrométrie de masse.

Obesity is a worldwide problem affecting both children and adults. Genetic, physiological, environmental, psychological, social and economic factors interact in varying degrees, influencing body weight and fat distribution and the progress of obesity. Moreover, some anthropogenic chemicals have proven to be endocrine disrupting chemicals (EDCs) with the potential to interfere with different actions of hormones in the body. EDCs may thereby disrupt homeostasis, modifying developmental, behavioral and immune functions in humans and animals, and also promoting adiposity. Because hormones generally act at low concentrations, small changes in the endocrine system may lead to extensive effects. Based on data from experimental and epidemiological studies this thesis elucidates the relationship between a large number of environmental contaminants and obesity. The experimental studies demonstrated that fructose supplementation in the drinking water resulted in unfavorable metabolic alterations such as a higher liver somatic index (LSI), an increase in plasma triglycerides and increased plasma levels of apo A-I. Fructose in combination with exposure to bisphenol A (BPA) increased liver fat content and plasma levels of apo A-I in juvenile female Fischer 344 rats. The experimental studies also showed that the retro-peritoneal fat, which in rats is a distinct fat depot easy to distinguish and dissect, correlated well with the measurements of total fat mass analyzed with MRI, and could therefore be used as a substitute for total fat mass in rats. The epidemiological studies showed that circulating levels of persistent organic pollutants (POPs) were related to fat mass measured by DXA. OCDD, HCB, TNC, DDE and the less chlorinated PCBs were positively related to fat mass, while the more highly chlorinated PCBs showed a negative association. Further, circulating levels of BPA were positively associated with levels of the hormones adiponectin and leptin, but negatively related with ghrelin, hormones which are involved in the regulation of hunger and satiety. However, serum BPA levels were not related to measures of fat mass in the elderly individuals in the PIVUS cohort. This thesis concludes that environmental contaminants such as BPA and POPs most likely are contributors, along with genetic, social and behavioral factors, to the development of obesity.