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This report was written in cooperation with Swedavia and aimed to examine and optimize the time before flight departure the chutes should open. Chutes are the last state before the baggage is transported to the plane. Currently, the chutes open two hours before departure. If baggage arrives while its corresponding chute is closed it will go to the Early Baggage System (EBS) where it will stay until two hours before departure. Afterward, the baggage will be transported to its corresponding chute. By reducing the time in the chute, the airport would utilize the EBS more efficiently and make the chutes available to more planes. To examine the resilience of the system it was modeled and simulated in Matlab. The parameters that were used to simulate the model were taken from a data set provided by Swedavia. The conclusion is that there's a possibility for higher utilization of the EBS by lowering the allotted times of chutes per flight and thereby freeing capacity in the makeup sector. This should be further investigated to see the effects of additional processes in the baggage handling system. / Denna rapport skrevs i samarbete med Swedavia och syftar till att undersöka och optimera tiden innan avgång som utsorteringsfickorna bör öppna. Utsorteringsfickorna är det sista stadiet innan bagaget transporteras till planet. I dagsläget öppnar utsorteringsfickorna två timmar innan avgång. Om bagaget anländer mer än två timmar innan avgång så hamnar väskan i Early Baggage System (EBS). Där kommer väskan att stanna tills det är två timmar till avgång och sedan transporteras till den korresponderande utsorteringsfickan. Genom att minska tiden som bagaget spenderar i utsorteringsfickan skulle flygplatsen kunna utnyttja EBS:en mer effektivt och göra utsorteringsfickor mer tillgängliga för fler plan.  För att undersöka systemets resiliens så modellerades systemet för att sedan simuleras i Matlab. Parametrarna som användes för att simulera modellen estimerades ifrån ett data set som Swedavia tillhandahöll. Slutsatsen från simuleringen var att det är möjligt att använda EBS mer effektivt genom att minska tiden i utsorteringsfickorna. Man bör därmed undersöka vidare för att se effekterna på det resterande bagagehanteringssystemet.

The thesis deals with the front part stiffness of modern vehicles, especially for the use in the field of forensic engineering in the traffic accident analysis. During the traffic accident analysis, an inquiry into the collision between vehicles is carried out which is an integral part of determining the energy loss of the vehicle at the impact, or more precisely the deformation energy expressed in the form of Energy Equivalent Speed (EES). In case of known stiffness of given part of the vehicle and based on the depth of deformation, it is possible to calculate the deformation energy, or more precisely EES corresponding with given damage of the vehicle. In the field of forensic engineering, the values of stiffness of individual vehicle components are not known and therefore, alternative methods are used to calculate the EES, they are outlined in the research part of this dissertation. However, the current methods of EES determination have some limitations when it comes to usability, and therefore, new EES calculation for the front part of the vehicle was designed in the research. It was based on the real crash test results using real stiffness characteristics of the front part of a vehicle. The front part of the vehicle is divided into individual areas and each of these parts is characterised by its own stiffness coefficient. The designed EES calculation can thus be also used for collisions with partial overlapping, taking into account the real stiffness of the damaged part of the vehicle, which was not possible with existing methods. In the research part of this dissertation, a computer programme to calculate deformation energy and EES was processed. It works with individual stiffness characteristics in given areas where the input data are entered by the user and include the depth of permanent front part deformation in individual areas, vehicle weight, the direction of an impact force and the friction coefficient on the contact surface. Considering the fact that the vehicle stiffness is also one of the control parameters at solving collisions in the PC-Crash simulation programme, which is used in forensic engineering practice for the analysis of a collision process, a supplementary computer programme was designed. Based on the above-mentioned input data, the supplementary programme can further calculate data for collision solving in the simulation programme, namely stiffness, restitution coefficient and the vehicle damage stated in the output report of the simulation programme. Based on these data, the expert thus has the opportunity to solve the collision of two vehicles in the simulation programme with as much preciseness as if it was a real collision.

Although researchers have been able to print small, simple, and avascular tissues, they have been unsuccessful in creating large, complex and vascularized organs. Printing large and complex three-dimensional tissues or organs involves utilizing a large quantity of cellular spheroids and layer-by-layer addition of spheroids. In this study, an in-house cell spheroid fabrication system was developed to produce cell spheroids with human liver cells (hepG2), human endothelial cells (hEC), human neural stem cells (hNSC), and induced pluripotent stem cells (iPSC). It offers the ability of fabricating uniform-sized spheroids repeatedly, which is essential when large and complex structures need to be produced. In order to test the spheroids’ ability to fuse, hEC spheroids were placed in line with one another and revealed successful fusion. Overall, the results indicate the in- house developed cell spheroid fabrication system can play a major role in bioprinting by providing researchers with uniform-sized spheroids in large quantities, consistently.

Ebselen (Ebs) is considered as a glutathione peroxidase (GPx) mimetic and primarily thought to function by scavenging intracellular reactive oxygen species (ROS). Previous to our work, Deng et al. (2010a) demonstrated complete block of ICl,swell with 15 microM Ebs following endothelin-1 (ET-1) induced activation of the current in cardiomyocytes. This block was presumed to take effect mainly via the quenching of ROS. Nonetheless, our work with DI TNC1 astrocytes strongly emphasizes that Ebs might function by an alternative mechanism based on its kinetic profile in blocking ICl,swell. Our experiments showed that 45 nM Ebs can fully block ICl,swell thus suggesting an apparent IC50 result, we predicted Ebs to possess a high kon with a low koff close to zero. As predicted, Ebs failed to washout in the timescale covered by our patch-clamp experiments. The block was also distal to H2O2, previously considered as the most proximate regulator of ICl,swell. And based on further evidence demonstrating irreversible block of ICl,swell distal to H2O2 with Ebs congeners, complete suppression of native ICl,swell with MTS reagents, and failure of Ebs to block ICl,swell from the cytosol, we concluded that Ebs and its congeners can covalently modify important –SH groups required for current activation while functioning as sulfhydryl reagents. Complete irreversible block of ICl,swell with 110 mM cell impermeant MTSES in native DI TNC1 astrocytes contrasts sharply to SWELL1 (Qiu et al., 2014) or LRRC8A (Voss et al., 2014), the latest molecular entity presumably responsible for ICl,swell, where 3.33 mM MTSES failed to demonstrate block of ICl,swell in the wild-type stably expressing SWELL1 (Qiu et al., 2014). Our data with Ebs, its congeners, and MTS reagents indicate the existence of a common extracellular binding site which involves a selenenylsulfide (Se-S) bond that critically modulates ICl,swell. We, therefore, synthesized a derivative of Ebs called ebselen-para-yne (Ebs-p-yne), which provided an even higher affinity for blocking ICl,swell with a presumed IC50 ~picomolar range. Ebs-p-yne is a promising novel molecule that may serve as a tag in identifying the molecular fingerprint ultimately responsible for ICl,swell. Furthermore, we can take advantage of click chemistry to ultimately pull out the channel or channel component which has remained elusive for greater than two decades.

Le cancer papillaire de la thyroïde (CPT) est la tumeur endocrine la plus fréquente. Des anomalies moléculaires activant la voie des MAPK (Mitogen-Activated Protein Kinases) sont identifiées, de façon mutuellement exclusive, dans environ 70% des cas. Il s’agit de réarrangements chromosomiques, le plus souvent de type RET/PTC (10%), de mutations ponctuelles activatrices des trois isoformes de l’oncogène RAS (H, N et K-RAS) (10%), ou de l’oncogène B-RAF (50%). La mutation « hot spot » B-RAFV600E est la plus fréquemment identifiée, elle est associée à une plus grande agressivité clinique (diagnostic à un stade tardif, risque de récidives et de décès accru). Ces évènements moléculaires ont pour conséquence commune l’activation de la voie des MAPK, se traduisant en aval par la phosphorylation de MEK (Mitogen-activated Extracellular signal-Regulated Kinase) puis de ERK (Extracellular signal-Regulated Kinase). Cette dernière est régulée négativement par des phosphatases, appartenant à la famille des Dual Specificity Phosphatases (DUSPs), d’expression ubiquitaire, et en particulier de deux phosphatases spécifiques de ERK, l’une cytoplasmique (DUSP6) et l’autre nucléaire (DUSP5). Nous avons fait l’hypothèse que ces phosphatases pouvaient être soit des gènes suppresseurs de tumeurs (leur perte d’expression conduisant à une augmentation de phosphorylation de ERK et une prolifération accrue), soit des marqueurs du degré d’activation de la voie MAPK dans le cadre d’une boucle de rétrocontrôle négatif. Ceci nous a conduits à analyser la régulation et l’expression de ces phosphatases dans trois modèles : la lignée cellulaire PCCL3 (thyroïde de rat), exprimant l’un des trois principaux oncogènes mutés dans les CPT (RET/PTC3 ou H-RASV12 ou B-RAFV600E) sous le contrôle d’un promoteur inductible par la doxycycline, des lignées cellulaires humaines dérivant de CPT et des CPT humains. (...) / Papillary thyroid cancer (PTC) is the most common endocrine malignancy. Mutually exclusive and activating alterations of the MAPK pathway (Mitogen-Activated Protein Kinases) are identified in 70% of cases. Common mutations found in PTCs are point mutation of the B-RAF (50%) and RAS genes (10%) as well as RET/PTC chromosomal rearrangements (10%). The hot spot B-RAFV600E mutation is the most frequently alteration identified and is connected with agressive clinical characteristics (high stage at diagnosis, high recurrence risk and death). These molecular events lead to constitutive activation of the MAPK pathway, resulting in MEK (Mitogen-activated Extracellular signal-Regulated Kinase) and ERK (Extracellular signal-Regulated Kinase) phosphorylation. ERK is negatively regulated by phosphatases and among them, Dual Specificity Phosphatases (DUSPs), ubiquitary expressed, in particular two ERK-specific phosphatases DUSP5 (nuclear) and DUSP6 (cytosolic). We hypothesized that these phosphatases could have tumor supressor properties (i.e. their loss would be associated with an increase in MAPK pathway activation) or may serve as a surrogate marker of MAPK pathway activation in the context of a negative feedback loop. We analysed regulation and expression of both phosphatases in 3 models: three PCCL3 cell lines (rat thyroid cells) expressing one of the most common oncogene identified in PTCs (RET/PTC3 or H-RASV12 or B-RAFV600E) under the control of a doxycycline-inducible promoter, human PTC-derived cell lines and human PTC. We demonstrated that MAPK pathway activation was correlated with induction of DUSP5 and DUSP6. These phosphatases are involved in a negative feedback loop that contributes to a tight regulation of phospho-ERK levels. DUSP5 and DUSP6 mRNA are overexpressed in human PTCs, especially in B-RAF mutated tumors suggesting a higher MAPK signaling output in these agressive PTCs. Silencing of DUSP5 and/or DUSP6 by small interfering RNA does not affect proliferation of human B-RAFV600E thyroid carcinoma-derived cell lines, suggesting the lack of tumor suppressor gene role. Compensatory changes in expression of DUSPs when a specific one is inactivated may explain this lack of effect. On the opposite, a DUSP6 pharmacological inhibitor induced a concentration dependent decrease in proliferation of human B-RAFV600E cells, suggesting « off-target » effect of this inhibitor. In a second part, we analysed the regulation of DUSP5 expression, which is a target of the MAPK pathway activation. We demonstrated, using pharmacological inhibitors, that DUSP5 is an early response gene, regulated mostly by the MAPK pathway, at the transcriptional level. Two contiguous CArG boxes that bind serum response factor (SRF) were found in a 1Kb promoter region, as well as several E twenty-six transcription factor family binding sites (EBS). These sites potentially bind Elk-1, a transcription factor activated by ERK1/2. Using wild type or mutated DUSP5 promoter reporters, we demonstrated that SRF plays a crucial role in serum induction of DUSP5 promoter activity, the proximal CArG box being important for SRF binding in vitro and in living cells. Moreover Elk-1 was bound in vitro to a promoter region containing the proximal CArG box and a putative EBS. Its specific binding to SRF was necessary to elicit promoter response to dominant positive Elk-VP16 and to enhance the response to serum stimulation. Altogether our results suggest that the MAPK pathway is more active in B-RAFV600E PTC than in PTC with other genetic alteration and could explain their clinical agressivity. DUSP5 and DUSP6, as well as phosphorylated MEK, are markers of activation of the MAPK pathway. Neither phosphatase has tumor suppressor properties in our thyroid cancer cell models. Our results suggest redundancy and functional compensation among DUSPs. (...)

L'ivoire d'éléphant est un matériau biologique composé de fibres de collagène (CF) à 30 % massique et de particules d'hydroxyapatite carbonatées et enrichies en Mg à 70 % massique (Mg-carb-HAP). Il présente une structure hiérarchique complexe de la macro à la nano-échelle. La relation entre le motif macroscopique de Schreger observé à la surface des sections transverses des défenses et la micro-morphologie de l'ivoire en 3D (réseau tubulaire et orientations secondaires des CF) a été établie. Les marqueurs chimiques (Ca, P, Mg, Sr) et structuraux (épaisseurs et organisation des particules de Mg-carb-HAP) témoins des processus de formation de l'ivoire ont été déterminés. La diagenèse précoce en milieu marin a ensuite été étudiée par une approche physico-chimique combinant les analyses MEB, PIXE/RBS-EBS et SAXS. Les mécanismes d'altération identifiés sont les adsorptions des ions du milieu extérieur (Cl, Sr, Fe, Cu) à la surface des défenses, les échanges entre les ions exogènes et endogènes de l'ivoire et l'augmentation de la cristallinité des Mg-carb-HAP. Bien qu'immergées dans le même environnement diagénétique, les trois défenses du site des Poulins présentent différents états d'altération. Un bon état de préservation macroscopique ne reflète pas forcément un bon état de conservation de la dentine à l'échelle moléculaire. Finalement, l'ancienne polychromie et la dorure d'origine des ivoires d'Arslan Tash (Syrie, 800 av. J.C.) ont été restituées par des analyses non-invasives par FX en plein champ et PIXE/RBS-EBS. Les couleurs identifiées sont: le bleu et le vert égyptiens (Cu), avec des teintes plus ou moins claires (Pb), le rouge et l'orange (Fe). / Elephant ivory is a biological material composed of collagen fibers (CF) at 30 wt. % and Mg-enriched carbonated hydroxyapatite particles at 70 wt. % (Mg-carb-HAP). It has a complex hierarchical structure from macro- down to nano-scale. The relationship between the macroscopic Schreger patterns observed on the surface of transverse sections of tuks and the 3D micro-morphology of ivory (tubular network and CF secondary orientations) has been established. Chemical (Ca, P, Mg, Sr) and structural markers (thickness and organization of particles Mg-carb-HAP) which control the formation of ivory have been determined. Early diagenesis in the marine environment was then studied by means of SEM, PIXE/RBS-EBS and SAXS analyses. Diagenetic mechanisms were identified, as ionic adsorptions from marine environment to the tusk surfaces, ionic substitutions between exogenous and endogenous ivory ions and increased crystallinity of Mg-carb-HAP. Different states of preservation were observed among three tusks coming from the same submarine archaeological site. Good macroscopic preservation states of the surface does not necessarily reflect good preservation states of the dentin at the molecular level. Finally, the former polychromy and gilding of ivories from Arslan Tash (Syria, 800 BC.) have been reconstructed by non-invasive FF-FX and PIXE/RBS-EBS analyses. Egyptian blue and green (Cu) with different shades (Pb), as well as red and orange (Fe) have been identified. The gilding technique consisted of applying a 2 µm thick gold leaf. Over time, these decorations altered ivory surfaces inducing, among others, the formation of Au nanoparticles derived from the weathering of the gold leafs.