It has been argued that for the management of complex issues such as sustainability, which transcend traditional policy sectors and require coordination between several different interests and actors, policymaking depends upon collaboration and integration processes between different sectors and tiers of government. The overall aim of this thesis is therefore to study how and why (or why not) policy integration processes are being developed in regional policymaking and what this means for the achievement of sustainable transport. The thesis consists of two separate qualitative case studies of policymaking in two Swedish regions, one representing a least likely case and the other a most likely case of policy integration. The focus has been on the organizational actors involved in policymaking processes for the regional transport system. For the general discussion the theoretical framework of policy integration, complemented by the analytical concepts of policy logics, organizational identities and boundary object are used. The findings are presented in four articles. An overall conclusion is that policy integration processes do not necessarily result in policy for sustainable transport. If policy integration becomes a goal in itself and the same as joint policy, it risks neglecting sustainable values and becoming the smallest common denominator that a number of actors can agree on. For developing sustainable transport solutions, collaboration for the coordination of policy may be beneficial, but the aim of such processes should not be joint policy. / För att beslut och riktlinjer ska kunna utformas så att de leder till lösningar av komplexa frågor, såsom hållbar utveckling, anses de behöva hanteras i samverkan mellan flertalet berörda sektorer och beslutsfattande nivåer. Det är dessa samverkansprocesser, beskrivna som integration under policy processer, som den här avhandlingen analyserar. Syftet är att studera om och hur integrerade regionala policyprocesser förekommer, hur de utvecklas samt deras betydelse för att åstadkomma ett hållbarare transportsystem. Detta undersöks genom kvalitativa fallstudier av två olika svenska regioner som representerar ett minst och ett mest troligt fall av integration av policy. Fallstudierna görs i regionerna Stockholms län och Västra Götalands län. Dessa två fall representerar dessutom två helt olika typer av regionala organisationer, vilket gör att de utgör underlag till, inte bara en diskussion om hållbara transporter, utan också om utvecklingen av den svenska regionala förvaltningsnivån. För analys används teori kring integration av policy och tre huvudsakliga analytiska begreppsansatser: policylogiker, organisationsidentiteter och gränsobjekt. Resultaten presenteras i fyra separata artiklar och dessa diskuteras tillsammans i den inledande kappan. I studien konstateras att integration av policysektorer och förvaltningsnivåer inte nödvändigtvis leder till transportlösningar som är mer hållbara. Beslut om en gemensam policy över sektorer och nivåer riskerar bli urvattnad eftersom det är många aktörer som ska komma överens. Samverkan för att samordna olika mål och intressen visar sig i huvudsak vara viktigt för att styra mot ett hållbart transportsystem, men det innebär inte att gemensam policy bör vara målet. Därutöver belyser studien hur olika organisationsformer på regional nivå påverkar regionala beslutsprocesser och hanteringen av hållbar transportutveckling.

Godstransporter ökar i takt med att utrikeshandeln växer, vilket bidrar till utsläpp av växthusgaser. Hållbar utveckling och miljö är viktiga faktorer i dagens samhälle vilket gör att det ställs krav på nya och innovativa transportlösningar. Ett alternativ att bemöta dessa krav är genom högkapacitetstranporter, som i dagsläget testas, utförs och forskas kring frekvent i Sverige. Högkapacitetstransport avser lastbilsfordon som är längre och/eller tyngre än dagens konventionella lastbil som har fått dispens att trafikera vissa zoner på det svenska vägnätet. Denna studie baseras på att utvärdera två intermodala transportlösningar för transport av gods på sträckan mellan Göteborgs hamn och Viared, Borås. Den ena transportlösningen baseras på högkapacitetstransport och den andra baseras på en järnvägsbaserad transportlösning. De alternativa transportlösningarna utvärderas med avseende på effektivitet omfattande transportkostnad, flexibilitet och tid samt miljöpåverkan omfattande koldioxidutsläpp genom en fallstudie.Slutsatserna har resulterat i att högkapacitetstransport genererar en högre transporteffektivitet jämfört med dagens dragbil som trafikerar den utpekade vägsträckan i fallstudien. I brist på material har inga konkreta kostnader kunnat beräknats för de olika transportlösningarna. Däremot kunde en slutsats dras kring att de alternativa transportlösningarna ej bör uppgå till en högre kostnad än vad dagens transportlösning genererar. Ur miljösynpunkt har koldioxidutsläpp för de olika transportlösningarna beräknats. Implementeras hela vägsträckan med högkapacitetstransport har resultatet gällande koldioxidutsläpp genererat i en minskning med 30,6 procent jämfört med dagens dragbil. Däremot visade studien att järnvägstransporten vinner miljömässiga fördelar då den jämfört med dragbilen som trafikerar vägsträckan idag reducerar utsläppen med 80,4 procent. Dock visar koldioxidutsläppet i den intermodala järnvägslösningen att vägtransporterna bidrar med 98 procent av utsläppen.Denna studie väger för- och nackdelar gentemot varandra för en implementering av högkapacitetstransport. Högkapacitetstransporten jämförs även gentemot järnvägens för- och nackdelar vid transport av containergods. Studien bidrar dessutom till att stärka och reflektera över tidigare publicerade studier som finns inom ämnet. / Freight transport increases as foreign trade grows, which contributes to greenhouse gas emission. Sustainable development and the environment are important factors in today's society, which imposes demands on new and innovative transport solutions. One way of meeting these demands is through high capacity transports, which are currently being tested, performed and researched frequently in Sweden. High capacity transport refers to truck vehicles that are longer and/or heavier than today's conventional truck that has been given the dispensation to traffic certain areas on the Swedish road network. This study is based on evaluating two intermodal transport solutions for the carriage of goods on the route between Port of Gothenburg and Viared, Borås. One transport solution is based on high capacity transport and the other is based on a rail-based transport solution. Through a case study, the alternative transport solutions are evaluated based on efficiency, including transportation costs, flexibility and time, and the environmental impact including CO2 emissions.The conclusions have resulted in high capacity transport generating a higher transport efficiency compared with today's truck that operates the designated road distance in the case study. In lack of material, no concrete costs have been calculated for the various transport solutions. On the other hand, a conclusion could be drawn that alternative transport solutions should not be at a higher cost than today's transport solution results in. From an environmental point of view, CO2 emissions for the various transport solutions have been calculated. Implementing the entire road haulage with high capacity transport has resulted in a reduction of CO2 emissions by 30,6 percent compared to the current truck. On the other hand, the study showed that rail transport wins environmental benefits, as compared to the truck that carries the route today, emissions reduce by 80,4 percent. However, CO2 emissions in the intermodal railroad solution show that road transport contributes 98 percent of emissions.This study weighs strengths and weaknesses towards each other for the implementation of high capacity transport on the designated route. High capacity transport is also set against the railroad with pros and cons of transporting container goods. The study also contributes to strengthening and reflecting the theory from previously published studies within the subject.

No description available.

Background: The business world is in constant change, which means that businesses must be able to adapt to these changes and constantly improve. The research field on how businesses should achieve a smart material supply is small, which emphasizes the importance of studying how a company can change its production layout to improve its internal logistics. Purpose: The purpose of this study is to increase the understanding of the production layout's importance for the material supply and the human aspect in production-related change work. Method: The study has used an abductive, exploratory approach to answer the purpose and the research questions. This means that studies of theories were done in parallel with studies of the empirical material at the case company PEPAB Produktionspartner AB. Theories from the literature and the empirical data have been analyzed and discussed in order to draw conclusions. Findings: The results are a description of the current internal material flow of four representative components and a description of employees´ attitude to change. By mapping the material flow, logistics flow factors and human related factors have been discovered.  Theoretical contribution: When changing the production starting from the logistic factors waiting, unnecessary inventory, unnecessary manufacturing, defects, unnecessary transportations and improperly designed production layout a better logistical flow and better material supply can be achieved. Combining this with a well-planned preparatory work with related training can make a successful change.  Practical contribution: This study contributes practically with a new production layout that enables a better logistical flow. It also emphasizes the importance of a well-thought-out and wellplanned preparatory work to maintain the staff´s sense of participation. / Bakgrund: Företagsvärlden är i konstant förändring vilket betyder att verksamheter måste anpassa sig till dessa förändringar och hela tiden förbättras. Forskningsfältet kring hur verksamheter ska uppnå en smart materialförsörjning är litet, vilket poängterar vikten av att studera hur ett företag kan förändra sin produktionslayout för att förbättra sin interna logistik. Syfte: Syftet med denna studie är att öka förståelsen av produktionslayoutens betydelse för materialförsörjningen samt den mänskliga aspekten i produktionsrelaterat förändringsarbete.  Metod: Studien har använt sig av en abduktiv, explorativ ansats för att besvara syfte och forskningsfrågor. Det betyder att studier av teorier gjorts parallellt med studier av det empiriska materialet på fallföretaget PEPAB Produktionspartner AB. Teorier från litteraturen och det empiriska materialet har analyserats och diskuterats för att slutligen dra slutsatser. Resultat: Resultatet av studien är en nulägesbeskrivning av det nuvarande interna materialflödet av fyra representativa komponenter och en nulägesbeskrivning över anställdas inställning till förändring. Genom kartläggning av materialflödet har logistiska flödesfaktorer och människorelaterade förändringsfaktorer framkommit.      Teoretiskt bidrag: Att vid förändring av produktionen utgå från de logistiska faktorerna väntan, onödig lagerhållning, onödig tillverkning, defekter, onödiga transporter och en felaktigt utformad produktionslayout kan ett bättre logistiskt flöde och en bättre materialförsörjning uppnås. Att kombinera detta med ett välplanerat förarbete med tillhörande utbildning kan leda till att förändringen blir lyckosam. Praktiskt bidrag: Denna studie bidrar praktiskt med en ny produktionslayout som leder till ett bättre logistiskt flöde, den poängterar även vikten av ett genomtänkt och välplanerat förarbete för att upprätthålla personalens känsla av delaktighet.

Ballast som byggnadsmaterial är viktigt för att utveckla vårt samhälle då det bland annat används som beståndsdel i asfalt och betong. Det är därför viktigt att bergkross som bryts i bergtäkter har en fungerande produktion för att möta efterfrågan men också för att Sverige ska kunna nå målet att vara klimatneutralt 2045. Syftet med arbetet är att minska miljöpåverkan i en bergtäkt från de interna transporterna och målet är att ta fram åtgärder för att uppnå detta. De metoder som har använts är litteraturstudier, beräkningar, simuleringar och intervjuer. Genom beräkningarna kunde ett resultat i detta arbete visa att en reducering med 88 % av koldioxidutsläpp kan göras genom att använda biodrivmedlet HVO Diesel 100. Det har med hjälp av intervjuer och litteraturstudier dessutom dragits en slutsats att framtidens bergtäkter antas producera bergkross med elektriska autonoma maskiner. / Ballast as a building material is important for developing our society as it’s used as a component in asphalt and concrete. Hence it is important that crushed rock extracted from a quarry have a functioning production to meet the demand that exists, but also because Sweden should be able to reach the goal of being climate neutral 2045. The purpose of this study is to reduce the environmental impact in a quarry from the internal transports and the aim is to bring out suggestions to achieve this. The methods that have been used are literature studies, calculations, simulations and interviews. Through the calculations, a result in this work showed that a reduction of 88 % of carbon dioxide emissions can be made by using the biofuel HVO Diesel 100. In addition, with the help of interviews and literature studies, conclusions have been drawn that the future quarries are believed to produce crushed rock with electric autonomous machines.

O diabetes mellitus tipo 2 (DM2) é caracterizado pela presença de resistência à insulina, a qual pode ser modulada pelo estrógeno, tanto em fêmeas como em machos. Nesse processo, o transportador de glicose GLUT4 (gene Slc2a4, solute carrier family 2 member 4) desempenha papel importante, pois aumento da expressão do GLUT4 melhora o controle glicêmico. Estradiol (E2) regula a expressão do Slc2a4 por meio do balanço dos efeitos contrários de seus receptores (ERs): ER-alfa estimula e ER-beta inibe a expressão. Efeitos transcricionais dos ERs envolvem a participação de co-reguladores, destacadamente o SP1 (transcription factor Sp1), potente estimulador do Slc2a4. Entretanto, o papel do SP1 na regulação do Slc2a4 mediada pelos ERs é desconhecido; e este foi o objetivo do presente estudo. Investigou-se adipócitos maduros 3T3-L1, tratados por 24 horas com E2, agonista de ER-alfa (PPT) ou agonista de ER-beta (DPN). Avaliou-se: a expressão gênica (RT-qPCR) de Slc2a4 e Sp1; o conteúdo (Western blotting) total de GLUT4 e o nuclear de ER-alfa/beta e SP1; a atividade de ligação do SP1 no Slc2a4 (ensaio de mobilidade eletroforética); e a formação de complexos SP1/ER-alfa (imunoprecipitação). Os resultados confirmaram que E2 aumenta a expressão de Slc2a4/GLUT4 pela ação preponderante do ER-alfa. O agonista PPT aumentou: o conteúdo nuclear de SP1, a interação SP1/ER-alfa e a atividade de ligação do SP1 no Slc2a4. O agonista DPN indicou que a ação repressora do ER-beta não envolve o SP1. Conclui-se que o efeito estimulador do ER-alfa na expressão do Slc2a4 envolve mecanismo de transativação gênica via SP1. Essas observações colocam a cooperação ER-alfa/SP1 como um novo alvo para o desenvolvimento de medidas terapêuticas para resistência à insulina e diabetes mellitus tipo 2 / Type 2 diabetes mellitus (T2DM) is characterized by insulin resistance, which can be modulated by estrogen in both females and males. In this process, the glucose transporter GLUT4 (solute carrier family 2 member 4 gene - Slc2a4) plays an important role, since increasing GLUT4 expression improves glycemic control. Estradiol (E2) regulates the expression of Slc2a4, by a mechanism in which estrogen receptors (ERs) play opposite effects: ER-alpha stimulates, whereas ER-beta inhibits the expression. Transcriptional effects of ERs involve co-regulators, notably the transcription factor SP1, a powerful enhancer of Slc2a4. However, the role of SP1 in the ERs-mediated regulation of Slc2a4 is unknown; and that was the aim of the present study. Differentiated adipocytes 3T3-L1 were treated (24 hours) with E2, ER-alpha agonist (PPT) or ER-beta agonist (DPN). It was analyzed: gene expression (RT-qPCR) of Slc2a4 and Sp1; total content o GLUT4 and nuclear content of ER-alpha/beta and SP1 (Western blotting); binding activity of SP1 into Slc2a4 promoter (electrophoretic mobility shift assay); and content of nuclear SP1/ER-alpha complexes (immunoprecipitation). Results confirmed that E2 increases the expression of Slc2a4/GLUT4, by the dominant effect of ER-alpha. The ER-alpha agonist PPT increased the nuclear content of SP1, the interaction of SP1/ER-alpha, and the binding activity of SP1 into the Slc2a4. The agonist DPN evinced that ER-beta activity does not involve the SP1. In conclusion, the enhancer effect of ER-alpha upon Slc2a4 gene expression involves a transactivation mechanism via SP1. This observation point outs the cooperation of ER-alpha/SP1 as a new target for the development of approaches to treat insulin resistance and T2DM

A nefropatia diabética (ND) é uma das principais causas de nefropatia crônica, o que torna o diabetes mellitus (DM) responsável por 44% da prevalência de doença renal crônica (DRC) no mundo. O papel do estresse oxidativo na patogênese da ND está bem estabelecido e genes pertencentes a vias pró- e antioxidantes são possíveis candidatos a conferirem susceptibilidade genética a essa e a outras complicações crônicas. Além do estresse oxidativo, o transporte intracelular de glicose, mediado por transportadores específicos, também parece exercer influência sobre a ND e outras complicações. O objetivo deste trabalho foi avaliar a associação entre ND e alguns polimorfismos de um único nucleotídeo (SNPs) em genes que codificam proteínas transportadoras de glicose (GLUT2 [SLC2A2]), proteínas pró-oxidantes (p22phox [CYBA] e NOX-2 [CYBB]) e proteínas antioxidantes (glutationa peroxidase-4 [GPX4] e catalase [CAT]) em uma coorte brasileira (n=453; 45,8% de pacientes com ND) e três coortes francesas (SURGENE [n=340; 17,7% de pacientes com ND na fase basal], GENEDIAB [n=313; 66,7% de pacientes com ND] e GENESIS [n=636; 49,7% de pacientes com ND]) de pacientes portadores de DM tipo 1. Os SNPs foram genotipados com o uso da técnica de reação em cadeia da polimerase (PCR) em tempo real e os resultados expressos em odds ratio (OR) ou hazard ratio (HR), com seus respectivos intervalos de confiança (IC), determinados em modelos ajustados de regressão logística politômica ou regressão de risco proporcional de Cox, respectivamente. A razão albumina/creatinina urinária (ACR) ou a taxa de excreção urinária de albumina (EUA) foram utilizadas para definir os estágios de ND e os pacientes foram classificados de acordo com a presença ou ausência de ND incipiente (ACR 30 - 300 mg/g de creatinina ou EUA 20 - 200 ?g/min ou 20 - 200 mg/L) e creatinina plasmática <1,7 mg/dL), ND estabilizada (ACR >300 mg/g de creatinina ou EUA > 200 ug/min ou > 200 mg/L e creatinina plasmática < 1,7 mg/dL ) ou ND avançada (ACR > 300 mg/g de creatinina ou EUA > 200 ug/min ou > 200 mg/L e creatinina plasmática > 1,7 mg/dL ou qualquer terapia de reposição renal) e também foram avaliadas associações dos SNPs com a taxa de filtração glomerular estimada (TFGe). O alelo raro A do SNP rs6610650 no gene CYBB foi associado com valores baixos de TFGe em mulheres na coorte brasileira e com a prevalência de ND estabilizada/avançada em mulheres da coorte francesa (OR 1,75; IC 95% 1,11 - 2,78; p=0,016). O alelo raro T do SNP rs713041 no gene GPX4 foi inversamente associado com a prevalência de ND estabilizada/avançada em homens na coorte brasileira (OR 0,30, IC95% 0,13 - 0,68, p=0,004) e com valores elevados de TFGe em homens na coorte francesa. O alelo raro A do SNP rs7947841 no gene CAT foi associado com a prevalência de ND incipiente (OR 2,79; IC95% 1,21 - 6,24; p=0,01) e ND estabilizada/avançada (OR 5,72; IC95% 1,62 - 22,03; p=0,007), bem como com a incidência de eventos renais, definidos como novos casos de microalbuminúria ou progressão para um estágio mais grave de ND durante o seguimento de estudo, na coorte SURGENE (HR 1,82; IC95% 1,13 - 2,81; p=0,01). O mesmo alelo de risco associou-se com a prevalência de ND incipiente (OR 3,13; IC95% 1,42 - 7,24; p=0,004) e com a incidência de insuficiência renal crônica terminal (IRCT) na coorte GENEDIAB (HR 2,11; IC95% 1,23 - 3,60; p=0,008) e com a prevalência de ND incipiente (OR 2,16; IC95% 1,14 - 4,10, p=0,02) e ND estabilizada/avançada (OR 2,71; IC95% 1,38 - 5,42; p=0,004) na coorte brasileira. O alelo raro T do SNP rs9932581 no gene CYBA foi inversamente associado com a prevalência de ND estabilizada/avançada (OR 0,60; IC95% 0,46 - 0,78; p=0,0001) e com valores mais baixos de TFGe nos pacientes de descendência europeia da coorte GENESIS/GENEDIAB. Este mesmo alelo foi associado com a incidência de eventos renais e de IRCT nas coortes SURGENE (HR 0,63; IC95% 0,46 - 0,86; p=0,003) e GENESIS/GENEDIAB (HR 0,51; IC95% 0,31 - 0,78; p=0,002), respectivamente. Entretanto estes resultados não foram replicados na coorte brasileira. O alelo raro T do SNP rs11924032 no gene SLC2A2 foi inversamente associado com a perda da TFGe ao logo do tempo (0,02%/ano vs 2,18%/ano para os pacientes portadores do genótipo GG; p=0,005), na coorte SURGENE. Este mesmo alelo foi inversamente associado com a incidência de IRCT nas coortes GENESIS/GENEDIAB (HR 0,53; IC95% 0,29 - 0,89; p=0,01). Os resultados observados para o gene SLC2A2 não forneceram fortes indícios para afirmarmos que este gene exerça um papel relevante no desenvolvimento da ND nos pacientes com DM tipo 1 nas coortes francesas estudadas. Em contrapartida, os SNPs nos genes que codificam as proteínas pró-oxidantes CYBA e CYBB e as proteínas antioxidantes GPX-4 e CAT foram capazes de modular o risco para doença renal em pacientes portadores de DM tipo 1, sendo que os SNPs presentes nos genes CYBB, GPX4 e CAT tiveram seus resultados replicados em coortes independentes, o que corrobora a importância destes genes e, consequentemente, do estresse oxidativo, na patogênese da ND / Diabetic nephropathy (DN) is a major cause of chronic nephropathy, with diabetes mellitus (DM) accounting for 44% of the prevalence of chronic kidney disease (CKD) in the world. The role of oxidative stress in the pathogenesis of DN is well established and genes belonging to pro- and antioxidant pathways are possible candidates to confer genetic susceptibility to this and other chronic complications. Besides oxidative stress, intracellular glucose transport mediated by specific transporters, also appears to influence DN and other complications. The aim of this study was to evaluate the association between DN and some single nucleotide polymorphisms (SNPs) present in genes encoding glucose transport proteins (GLUT2 [SLC2A2]), pro- (p22phox [CYBA] and NOX-2 [CYBB]) and antioxidants (glutathione peroxidase-4 [GPX4] and catalase [CAT]) proteins, in a Brazilian cohort [n= 453; 45.8% f patients with DN], and three French cohorts (SURGENE [n=340; 17.7% of patients with DN at baseline], GENEDIAB [n=313; 66.7% of patients with DN], and GENESIS [n=636; 49.7% of patients with DN]) of patients with type 1 DM. The SNPs were genotyped using the technique of real time polymerase chain reaction (PCR) and results expressed as odds ratio (OR) and hazard ratio (HR), with their respectively 95% confidence intervals (CI), determined by adjusted models of polytomic logistic regression and Cox proportional hazard regression, respectively. The albumin/creatinine ratio (ACR) or the urinary albumin excretion (UAE) rate were used to define the DN stages and the patients were classified according to the presence or absence of incipient DN (ACR 30 - 300 mg/g of creatinine or UAE 20 - 200 ug/min or 20 - 200 mg/L) and plasmatic creatinine < 1,7 mg/dL), established DN (ACR > 300 mg/g of creatinine or EUA > 200 ug/min or > 200 mg/L and plasmatic creatinine <1,7 mg/dL) or advanced DN (ACR >300 mg/g of creatinine or UAE > 200 ug/min or > 200 mg/L and plasmatic creatinine > 1,7 mg/dL or any renal replacement therapy). Associations for the estimated glomerular filtration rate (eGFR) were also evaluated. The rare allele A of the SNP rs6610650 in CYBB gene was associated with low values of eGFR in women in the Brazilian cohort and with the prevalence of established/advanced DN in women in the French cohort (OR 1.75, 95%CI 1.11 - 2.78, p=0.016). The rare allele T of the SNP rs713041 in GPX4 gene was inversely associated with the prevalence of established/advanced DN in men in the Brazilian cohort (OR 0.30, 95%CI 0.13 - 0.68, p=0.004) and with higher values of eGFR in men in the French cohort. The rare allele A of the SNP rs7947841 in CAT gene was associated with the prevalence of incipient DN (OR 2.79, 95%CI 1.21 - 6.24, p=0.01) and established/advanced DN (OR 5.72; 95%CI 1.62 - 22.03, p=0.007) as well as the incidence of renal events, defined as new cases of microalbuminuria or progression to a more severe stage during the follow-up study, in SURGENE cohort (HR 1.82, 95%CI 1.13 - 2.81, p=0.01). The same risk allele was associated with the prevalence of incipient DN (OR 3.13, 95%CI 1.42 - 7.24, p=0.004), the incidence of end-stage renal disease (ESRD) in the cohort GENEDIAB (HR 2.11, 95%CI 1.23 - 3.60, p=0.008) and with the prevalence of incipient DN (OR 2.16, 95%CI 1.14 - 4.10, p=0.02) and established/advanced DN (OR 2.71, 95%CI 1.38 - 5.42, p=0.004) in the Brazilian cohort. The rare T allele of the SNP rs9932581 in CYBA gene was inversely associated with the prevalence of established/advanced DN (OR: 0.60, 95%CI: 0.46 - .78, p=0.0001) and associated with lower values of eGFR in patients of GENESIS/GENEDIAB cohort. The same allele was inversely associated with the incidence of renal events and ESRD in SURGENE (HR 0.63, 95%CI 0.46 - 0.86, p=0.003) and GENESIS/GENEDIAB (HR 0.51, 95%CI 0.31 - 0.78, p=0.002) cohorts. However, these results were not replicated in the Brazilian cohort. The rare T allele of the SNP rs11924032 in SLC2A2 gene was inversely associated with the loss of eGFR during the follow-up (0.02%/year vs. 2.18%/year for patients with the GG genotype, p=0.005) in the SURGENE cohort. The same allele was inversely associated with the incidence of ESRD in the GENESIS/GENEDIAB cohorts (HR 0.53, 95%CI 0.29 - 0.89, p=0.01). The results observed for the SLC2A2 gene, in this study, did not provide strong evidence to state that this gene exerts a relevant role in the development of DN in patients with type 1 DM in the studied cohorts. However, SNPs in genes encoding the pro-oxidant proteins CYBA and CYBB, and the antioxidants proteins GPX-4 and CAT were able to modulate the risk of renal disease in patients with type 1 DM. The studied SNPs in CYBB, GPX4 and CAT genes had their results replicated in independent cohorts, which confirms the importance of these genes and, hence, of the oxidative stress in the pathogenesis of DN

Der Monocarboxylattransporter 8 (Mct8) ist ein spezifischer Schilddrüsenhormon (SDH)-Transporter. MCT8-Mutationen führen zu einer psychomotorischen Retardierung in Kombination mit abnormalen SDH-Serumkonzentrationen. Das konstitutiv Mct8-defiziente Mausmodell repliziert den endokrinologischen, jedoch nicht den humanen neurologischen Phänotyp. Um die Hypothese eines stark beeinträchtigten T3-Transportes speziell in Neuronen als Ursache zu untersuchen, wurde das Neuron-spezifische Mct8-defiziente Mausmodell (CamK-Cre;Mct8fl/fl) generiert. Neben einer funktionalen, Mct8-exprimierenden Blut-Hirn-Schranke liegt eine funktionale Hypophysen-Hypothalamus-Schilddrüsen Achse vor. NMR-Analysen des zerebralen Energiestoffwechsels von CamK-Cre;Mct8fl/fl-Mäusen zeigen nach [1-13C] Glukoseinfusion verringerte Laktatintensitäten sowie eine reduzierte Laktatdehydrogenase-Aktivität. Zudem sind Astrozyten-spezifische Transporter und Enzyme des Neurotransmitterstoffwechsels und deren Biosynthese in ihrer Genexpression reduziert. Somit führt der neuronale Mct8-Verlust zu einem verlangsamten zerebralen Metabolismus sowie einer reduzierten neuronalen Aktivität. Die Rolle von Mct8 im Energiestoffwechsel wurde außerdem in primären Mct8-defizienten Astrozyten- und Neuronkulturen mittels Seahorse Flux Analyzer untersucht. In Mct8-defizienten Neuronen kommt es zu einer verringerten SDH-Aufnahme, was in einer verringerten Expression von OXPHOS-relevanten Proteinen sowie in einer verringerten Sauerstoffverbrauchsrate resultiert. Somit stützen die in vitro Daten die des CamK-Cre;Mct8fl/fl-Mausmodelles bezüglich einer reduzierten neuronalen Aktivität sowie eines verlangsamten zerebralen Stoffwechsels. Zusammenfassend zeigen die Ergebnisse, dass grundlegende Mechanismen des zerebralen Stoffwechsels bei neuronaler Mct8-Defizienz beeinträchtigt sind und die Rolle von Mct8 mit Hilfe weiterer konditioneller Mausmodelle (Astrozyten-spezifisch) und primären Ko-Kulturmodellen untersucht werden muss. / The monocarboxylate transporter 8 (Mct8) is the most specific thyroid hormone (TH) transporter. Mutations lead to a severe form of psychomotor retardation in combination with abnormal TH concentrations in sera. The global Mct8-deficient mouse model was intensively studied and it replicates the endocrine, but not the human neurological phenotype. To test the hypothesis, that a disturbed uptake of T3 especially into neurons is responsible for the phenotype, we generated a neuron-specific Mct8-deficient mouse model (CamK-Cre;Mct8fl/fl). CamK-Cre;Mct8fl/fl mice exhibit a functional Mct8-expressing blood-brain-barrier and a functional hypothalamus pituitary thyroid axis. NMR analyses of the cerebral energy metabolism of CamK-Cre;Mct8fl/fl mice after [1-13C] glucose injection revealed less enrichment of lactate and a reduced lactate dehydrogenase activity. Moreover, especially astrocyte-specific expressed transporter and enzymes of neurotransmitter metabolism and their biosynthesis are significantly reduced in comparison to control mice. These results point to a decelerated cerebral metabolism as well as a reduced neuronal activity caused by the neuronal loss of Mct8. In addition, we studied the impact of Mct8 on the energy metabolism in primary wildtype and Mct8-deficient astrocyte and neuron cultures by use of the Seahorse Flux Analyzer. Mct8-deficient neurons show a reduced uptake of TH, which results in a reduced expression of OXPHOS relevant proteins as well as a reduced oxygen consumption rate. Therefore, the in vitro raised data provide the observed changes of the CamK-Cre;Mct8fl/fl mice regarding a reduced synaptic activity as well as a reduced cerebral metabolism. Taken together, the data clearly shows that basic mechanisms of the cerebral metabolism are hampered in neuronal Mct8 deficiency. The role of Mct8 in this context needs further analyses with the help of conditional mouse models (astrocyte-specific) and primary co-culture models.

La dystrophie myotonique de type 1 (DM1), ou maladie de Steinert, est une maladie qui touche plusieurs tissus, dont le système nerveux central (SNC). L’atteinte neurologique est variable et inclut des troubles de la fonction exécutive, des changements de comportement et une hypersomnolence dans la forme adulte, ainsi qu’une déficience intellectuelle marquée dans la forme congénitale. Dans leur ensemble, les symptômes neurologiques ont un fort impact sur le parcours académique, professionnel et les interactions sociales. Aujourd’hui aucune thérapie n’existe pour cette maladie. La DM1 est due à une expansion anormale d’un triplet CTG non-codant dans le gène DMPK. Les ARN messagers DMPK, porteurs de l’expansion, s’accumulent dans le noyau des cellules (sous forme de foci) et perturbent la localisation et la fonction de protéines de liaison à l’ARN, notamment des familles MBNL et CELF, ce qui entraîne des défauts d’épissage alternatif, d’expression, de polyadenylation et de localisation d’autres ARN cibles. Malgré le progrès récent dans la compréhension des mécanismes de la maladie, les aspects cellulaires et moléculaires de l’atteinte neurologique restent méconnus: nous ne connaissons ni la contribution de chaque type cellulaire du cerveau, ni les voies moléculaires spécifiquement dérégulées dans chaque type cellulaire. L’objectif de ma thèse a été de répondre à ces deux questions importantes en utilisant un modèle de souris transgéniques et des cellules primaires dérivées de celui-ci. Pour mon projet, j’ai utilisé les souris DMSXL générées par mon laboratoire. Ces souris reproduisent des caractéristiques importantes de la DM1, notamment l’accumulation des ARN toxiques et la dérégulation de l’épissage alternatif dans plusieurs tissus. L’impacte fonctionnel des transcrits DMPK toxiques dans le SNC des souris DMSXL se traduit par des problèmes comportementaux et cognitifs et par des défauts de la plasticité synaptique. Afin d’identifier les mécanismes moléculaires associés à ces anomalies, une étude protéomique globale a montré une dérégulation de protéines neuronales et astrocytaires dans le cerveau des souris DMSXL. De plus, l’étude de la distribution des foci d’ARN dans les cerveaux des souris et des patients a montré un contenu plus élevé dans les astrocytes par rapport aux neurones. Ensemble, ces résultats suggèrent une contribution à la fois neuronale et gliale dans la neuropathogenèse de la DM1. L’étude protéomique globale des cerveaux des souris DMSXL, a aussi montré des défauts de protéines synaptiques spécifiques des neurones, que nous avons par la suite validés dans le cerveau des patients. SYN1 est hyperphosphorylée d’une façon CELF-dépendante et RAB3A est surexprimé en réponse à l’inactivation de MBNL1. Les protéines MBNL et CELF régulent l’épissage alternatif d’un groupe de transcrits au cours du développement, et leur dérégulation dans la DM1 entraîne l’expression anormale d’isoformes d’épissage embryonnaires dans le tissu adulte. Dans ce contexte, j’ai étudié si les défauts des protéines RAB3A et SYN1 sont associés à une dérégulation d’épissage, et si les anomalies des protéines synaptiques identifiées dans la DM1 reproduisent des évènements embryonnaires de la régulation de RAB3A et SYN1. Mes résultats indiquent que les défauts de ces protéines dans les cerveaux adultes ne sont pas dus à une altération de l’épissage alternatif des transcrits et ne recréent pas des évènements embryonnaires. La neuropathogenèse de la DM1 va, donc, au delà de la dérégulation de l’épissage et d’autres voies moléculaires restent à explorer dans les cerveaux DM1. Afin d’identifier des sous-populations cellulaires susceptibles à l’accumulation des ARN toxiques, nous avons étudié la distribution des foci dans plusieurs régions cérébrales. (...) / Myotonic dystrophy type 1 (DM1) is a severe disorder that affects many tissues, including the central nervous system (CNS). The degree of brain impairment ranges from executive dysfunction, attention deficits, low processing speed, behavioural changes and hypersomnia in the adult form, to pronounced intellectual disability in the congenital cases. The neurological manifestations have a tremendous impact on the academic, professional, social and emotional aspects of daily life. Today there is no cure for this devastating condition. DM1 is caused by the abnormal expansion of a CTG trinucleotide repeat in the 3’UTR of the DMPK gene. Expanded DMPK transcripts accumulate in RNA aggregates (or foci) in the nucleus of DM1 cells, disrupting the activity of important RNA-binding proteins, like the MBNL and CELF families, and leading to abnormalities in alternative splicing, gene expression, RNA polyadenylation, localisation and translation. In spite of recent progress, fundamental gaps in our understanding of the molecular and cellular mechanisms behind the neurological manifestations still exist: we do not know the contribution of each cell type of the CNS to brain dysfunction, or the molecular pathways specifically deregulated in response to the CTG expansion. The aim of my PhD project has been to gain insight into these two important questions using a relevant transgenic mouse model of DM1 and cell cultures derived thereof. In my studies I used the DMSXL mice, previously generated in my host laboratory. The DMSXL mice express expanded DMPK mRNA with more than 1,000 CTG repeats. They recreate relevant DM1 features, such as RNA foci and missplicing in multiple tissues. The functional impact of expanded DMPK transcripts in the CNS of DMSXL mice translates into behavioural and cognitive abnormalities and defective synaptic plasticity. To identify the molecular mechanisms behind these abnormalities, a global proteomics analysis revealed changes in both neuron-specific and glial-specific proteins in DMSXL brain. We also investigated RNA foci in DMSXL and human DM1 brains and found non-homogenous distribution between cell types, with a higher foci content in astrocytes relative to neurons. Together these results suggest that both neuronal and glial defects contribute to DM1 neuropathogenesis. The global proteomics analysis of DMSXL brains also identified abnormalities in neuronal synaptic proteins that we have validated in human brain samples. SYN1 is hyperphosphorilated in a CELF-dependent manner while RAB3A is upregulated in association with MBNL1 depletion. CELF and MBNL proteins regulate the alternative splicing of a subset of transcripts throughout development, and their deregulation in DM1 leads to abnormal expression of fetal splicing isoforms in adult DM1 brains. In this context, I have studied if RAB3A and SYN1 deregulations observed in adult brains are associated with splicing abnormalities or if they recreated embryonic expression and phosphorylation events. My results indicate that the synaptic proteins abnormalities observed in adult DMSXL brains are not caused by defective alternative splicing and do not recreate embryonic events. Thus, DM1 neuropathogenesis goes beyond missplicing and other molecular pathways must be explored in DM1 brains. To better understand the cellular sub-populations susceptible of accumulating toxic RNA foci we have studied foci distribution in different brain regions. We identified pronounced accumulation of toxic RNAs in Bergman astrocytes of DMSXL mice cerebellum and DM1 patients, associated with neuronal hyperactivity of Purkinje cells. A quantitative proteomics analysis revealed a significant downregulation of GLT1 – a glial glutamate transporter expressed by the Bergmann cell in the cerebellum. I have confirmed the GLT1 downregulation in other brain regions of mouse and human brain. (...)

Repetitive Störungen manifestieren sich als Leitsymptom in der Zwangsstörung und dem Tourette-Syndrom. Die Symptome werden als enthemmte Stereotypien eines desinhibierten Basalganglien-thalamo-kortikalen (BGTC) Regelkreises verstanden. Überdies wird als neurochemisches Korrelat ein dysregulatives Dopamin (DA)-System innerhalb dieser Kerngebiete nahegelegt, welches über ein überaktives Dopamintransporter (DAT)-System erklärt werden könnte. In der Induktion repetitiver Erkrankungen ist die Interaktion des BGTC Regelkreises und des DA-Systems dennoch unklar. In der vorliegenden Arbeit wurden daher anhand von zwei Pathologiemodellen (Ratte) mit unterschiedlich induzierter Dysregulation des DA-Systems (transgen versus pharmakologisch) die dysfunktionalen Einheiten im BGTC Regelkreises vergleichend untersucht. Im transgenen Modell führte die zentralnervöse DAT-Überexpression: (1) zu einer verstärkten Genexpression des vesikulären Monoamintransporter 2 (VMAT2) sowie des DA-Rezeptors 1 und DA–Rezeptors 2 (DRD1, DRD2), (2) zu einem reduzierten DA-Spiegel mit erhöhter DA-Umsatzrate und veränderten serotonergen- und GABAergen-System, und (3) zu perserverativen Verhalten. Im Gegensatz dazu zeigte die chronische Applikation mit dem D2-Agonisten Quinpirol im pharmakologischen Modell: (1) eine Reduktion des DAT, VMAT2 und DRD2, (2) eine reduzierte DA-Umsatzrate und (3) zwanghaftes Kontrollverhalten. Die Ergebnisse legen nahe, dass die unterschiedlichen klinischen Subtypen der Zwangsstörung unterschiedlichen neurobiologischen Veränderungen zugrunde liegen könnten. Ferner bietet das hier vorgestellte transgene Modell erfolgsversprechende Ansatzpunkte um als neues valides Tiermodell der repetitiven Störungen etabliert zu werden. / Repetitive disorders manifest as the cardinal symptom in obsessive-compulsive disorder and Tourette syndrome. The symptoms are understood as disinhibited stereotypies of a basal ganglia-thalamo-cortical (BGTC) circuit. Furthermore, it is suggested that a dysregulated dopamine (DA) system within this circuit is the underlying neurochemical correlate which could be explained by an overactive dopamine transporter (DAT). At this point, it is still unclear how the BGTC circuit and the DA system interact in the induction of repetitive disorders. Therefore we investigated the dysfunctional unities within the BGTC circuit by comparing two pathological rat models (transgenic versus pharmacologic) with different induced dopaminergic dysregulation. The DAT overexpressing rat model showed: (1) increased gene expression of the vesicular monoamine transporter 2 (VMAT2), DA receptor D1 (DRD1) and DA receptor D2 (DRD2), (2) lower levels of DA with an increased DA metabolism and alterations in the serotonin- and GABA system, and (3) perseverative behavior. In contrast, the chronic application of the D2 receptor agonist quinpirole resulted in the pharmacologic model in: (1) lower gene expressions of the DAT, VMAT2 and DRD2, (2) reduced DA-turnover and (3) compulsive control behavior. These results suggest that different clinical subtypes of obsessive-compulsive disorder caused by different neurobiological alterations. In addition, the presented transgenic model provides the opportunity to be established as a new valid animal model of repetitive disorders.