Les petites protéines G Arf ont besoin d'un facteur d'échange nucléotidique (GEF) afin de passer de leur forme inactive liée au GDP à leur forme active liée au GTP. GBF1 est la GEF pour Arf1 qui assure, notamment, le recrutement du complexe manteau COPI impliqué dans le transport entre le Golgi précoce et le réticulum endoplasmique. Il a été récemment montré que GBF1 est impliqué dans la livraison de l'Adipose TriGlycéride Lipase (ATGL) sur les corps lipidiques (LDs). ATGL est une enzyme qui catalyse l'hydrolyse des triglycérides en diglycérides. Les travaux présentés dans cette thèse ont eu pour objectif d'étudier et de caractériser l'interaction entre GBF1 et la lipase ATGL. Par des expériences de co-immunoprécipitation dans les cellules de mammifère, les domaines des deux protéines impliquées dans l'interaction ont été identifiés. Par des expériences de pulldown utilisant les protéines exprimées chez E. coli, j'ai montré que ces interactions sont directes. Afin d'approfondir l'étude de l'interaction entre GBF1 et ATGL, j'ai construit des outils permettant l'étude biochimique de GBF1 en purifiant plusieurs de ses domaines. J'ai tout d'abord cherché à mettre au point un test d'activité pour GBF1 afin de tester l'influence de protéines partenaires, dont ATGL, sur son activité. Malgré la purification de différents fragments de GBF1 contenant le domaine Sec7, aucun n'a présenté une activité avec Arf1Δ17 en solution. Le domaine N-terminal de la protéine, avec et sans une mutation empêchant une interaction intramoléculaire, ainsi que les domaines HDS1 et HDS2 de GBF1 ont également été purifiés / Small G proteins Arf require assistance from a Guanine nucleotide exchange factor (GEF) in order to switch between GDP- and GTP-bound forms. GBF1 is the Arf1 GEF that mediates COPI coat complex recruitment to early secretory pathway membranes. COPI is a protein that coats vesicles transporting proteins from the cis side of the Golgi complex back to the rough endoplasmic reticulum. GBF1 was recently shown to mediate delivery of Adipose TriGlyceride Lipase (ATGL) to the surface of lipid droplets (LDs). ATGL is an enzyme catalyzing the initial step in triglyceride hydrolysis in LDs. Thus, the aim of this work was to study interactions between GBF1 and ATGL. By co-immunoprecipitation experiments in mammalian cells, the domains of two proteins involved in the interaction have been identified. By pulldown assays using proteins expressed in bacteria, I showed that these interactions are direct. To further study of the GBF1-ATGL interaction, I developed tools for the biochemical study of GBF1, by purifying several of its domains. I first tried to develop a kinetic essay for GBF1 to test the influence of interacting partners, including ATGL, on its activity. Despite the purification of various GBF1 fragments containing the Sec7 domain, none have activity with Arf1Δ17 in solution. The N-terminal domain of the protein, with and without a mutation disrupting an intramolecular interaction, and the HDS1 and HDS2 domains of GBF1 were also purified.

Background: Compare Goldmann applanation tonometer (GAT) prism and correcting applanation tonometry surface (CATS) prism to intracameral intraocular pressure (IOP), in vivo and in vitro. Methods: Pressure transducer intracameral IOP was measured on fifty-eight (58) eyes undergoing cataract surgery and the IOP was modulated manometrically to 10, 20, and 40 mmHg. Simultaneously, IOP was measured using a Perkins tonometer with a standard GAT prism and a CATS prism at each of the intracameral pressures. Statistical comparison was made between true intracameral pressures and the two prism measurements. Differences between the two prism measurements were correlated to central corneal thickness (CCT) and corneal resistance factor (CRF). Human cadaver eyes were used to assess measurement repeatability. Results: The CATS tonometer prism measured closer to true intracameral IOP than the GAT prism by 1.7+/-2.7 mmHg across all pressures and corneal properties. The difference in CATS and GAT measurements was greater in thin CCT corneas (2.7+/-1.9 mmHg) and low resistance (CRF) corneas (2.8+/-2.1 mmHg). The difference in prisms was negligible at high CCT and CRF values. No difference was seen in measurement repeatability between the two prisms. Conclusion: A CATS prism in Goldmann tonometer armatures significantly improve the accuracy of IOP measurement compared to true intracameral pressure across a physiologic range of IOP values. The CATS prism is significantly more accurate compared to the GAT prism in thin and less rigid corneas. The in vivo intracameral study validates mathematical models and clinical findings in IOP measurement between the GAT and CATS prisms.

Background: Goldmann applanation tonometry (GAT) error relative to intracameral intraocular pressure (IOP) has not been examined comparatively in both human cadaver eyes and in live human eyes. Futhermore, correlations to biomechanical corneal properties and positional changes have not been examined directly to intracameral IOP and GAT IOP. Methods: Intracameral IOP was measured via pressure transducer on fifty-eight (58) eyes undergoing cataract surgery and the IOP was modulated manometrically on each patient alternately to 10, 20, and 40 mmHg. IOP was measured using a Perkins tonometer in the supine position on 58 eyes and upright on a subset of 8 eyes. Twenty one (21) fresh human cadaver globes were Intracamerally IOP adjusted and measured via pressure transducer. Intracameral IOP ranged between 5 and 60 mmHg. IOP was measured in the upright position with a Goldmann Applanation Tonometer (GAT) and supine position with a Perkins tonometer. Central corneal thickness (CCT) was also measured. Results: The Goldmann-type tonometer error measured on live human eyes was 5.2 +/- 1.6 mmHg lower than intracameral IOP in the upright position and 7.9 +/- 2.3 mmHg lower in the supine position (p <.05). CCT also indicated a sloped correlation to error (correlation coeff. = 0.18). Cadaver eye IOP measurements were 3.1+/-2. 5 mmHg lower than intracameral IOP in the upright position and 5.4+/- 3.1 mmHg in the supine position (p <.05). Conclusion: Goldmann IOP measures significantly lower than true intracameral IOP by approximately 3 mmHg in vitro and 5 mmHg in vivo. The Goldmann IOP error is increased an additional 2.8 mmHg lower in the supine position. CCT appears to significantly affect the error by up to 4 mmHg over the sample size.

In Sweden there are many ice halls and ice halls climate differ from the usual buildings' climate, because temperatures are lower indoors than outdoors during the summer months. This results in a higher vapor content outdoors compared with indoors during the course of the year. Moisture transport takes place from the outside towards the ice halls, which is contrary to what happens in a common building. During the critical period, the vapor transport direction is inward. When the hot air hits the cold surface of the wall structure, it leads to a risk of condensation in the wall that leads to moisture damage. Moisture damage can impair the wall structure and lead to unwanted consequences. The purpose of this study was to dimension a number of walls that meet climate conditions, which are done by dynamic humidity calculation in WUFI Pro 5.1 with adapted climate data and real climate data and then mold analysis in WUFI Bio. The aim of the bachelor thesis work was that the results from this study can then be used as a guide to the future building of ice skating walls. The results from WUFI Pro 5.1 and WUFI Bio have shown that a number of walls designed during this master thesis are judged to work as well as revealed how an ice hallway wall should not be constructed. The conclusion shows that climate conditions control the design of the wall structure. / I Sverige finns det många ishallar och ishallsklimatet skiljer sig från vanliga byggnaders klimat, detta på grund av att temperaturen är lägre inomhus än utomhus under sommarhalvåret. Detta resulterar i en högre ånghalt utomhus jämfört med inomhus under en del av året. Fukttransporten sker då utifrån och in mot ishallen, det vill säga tvärtemot vad som sker i en vanlig byggnad. Under den kritiska perioden är ångtransportsriktningen inåt. När den varma luften träffar den kalla ytan i väggkonstruktionen, medför det en risk för kondens i väggen som leder till fuktskador. Fuktskador kan försämra väggkonstruktionen och leda till oönskade konsekvenser som exempelvis mögelpåväxt och rötskador. Syftet med studien har varit att dimensionera ett antal väggar som klarar av klimatförutsättningarna, detta gjordes genom dynamisk fuktberäkning i WUFI Pro 5.1 med anpassade klimatdata samt verkliga klimatdata och sedan mögelanalys i WUFI Bio. Målet med examensarbetet var att resultaten från denna studie sedan kan komma att användas som en vägledning till framtida byggen av ishallsväggar. Resultaten från WUFI Pro 5.1 och WUFI Bio har visat att ett antal väggar som konstruerats under detta examensarbete bedöms att fungera samt kommit fram till hur en ishallsvägg inte skall konstrueras. Slutsatesen visar att klimatförutsättningarna styr dimensioneringen av väggkonstruktionen.

The topic of this diploma thesis is systems of energy efficient buildings with a focus on the building envelope. The first theoretical part describes the quantities that we consider when designing the building envelope. The second half of the theoretical part is with reference to the experimental part of the thesis focused on a special type of vertical perimeter structure, namely walls with live vegetation installed from the outside of the structure. The second, practical part of this work, deals with the design of heating and hot water supply for a multifunctional building. The design is elaborated in two variants - heating by means of radiators in the first variant and in the second variant by means of floor heating for residential units of the solved building. The third part define an experiment, which had the task of comparing the measured quantities on two types of vertical perimeter structures, namely a structure with growing greenery from the outside and an identical structure only without a layer of greenery. At the same time, the task was to check the possible influence of the applied greenery on heat losses, and thus the design of the heating system.