Today’s heavy duty vehicles are equipped with safety and comfort systems, e.g. ABS and ESP, which totally or partly take over the vehicle in certain risk situations. When these systems become more and more autonomous more robust positioning is needed. In the right conditions the GPS system provides precise and robust positioning. However, in harsh environments, e.g. dense urban areas and in dense forests, the GPS signals may be affected by multipaths, which means that the signals are reflected on their way from the satellites to the receiver. This can cause large errors in the positioning and thus can give rise to devastating effects for autonomous systems. This thesis evaluate different methods to enhance a low cost GPS in harsh environments, with focus on mitigating multipaths. Mainly there are four different methods: Regular Unscented Kalman filter, probabilistic multipath mitigation, Unscented Kalman filter with vehicle sensor input and probabilistic multipath mitigation with vehicle sensor input. The algorithms will be tested and validated on real data from both dense forest areas and dense urban areas. The results show that the positioning is enhanced, in particular when integrating the vehicle sensors, compared to a low cost GPS.

Cell-cell junctions are essential for tissue homeostasis. Prominent among these junctions are adherens junctions and tight junctions. Adherens junctions mediate adhesion between adjacent cells while tight junctions are responsible for establishing apical-basolateral polarity and limiting paracellular permeability. Loss or disruption of either adherens junctions or tight junctions leads to a myriad of disease states, thus these junctions need to be tightly regulated to prevent dysfunction. A unique property of tight junctions is their dependence on adherens junctions for proper assembly and maintenance. Loss or disruption of adherens junction leads to abnormal tight junctions. Understanding the mechanisms that mediate tight junction coupling to adherens junctions is important for treating diseases that arise from disrupted cell-cell junctions. Currently, two controversial models exist for how tight junctions are coupled to adherens junctions. In the first model, the adherens junction protein α-catenin is critical for tight junction assembly. The second model suggests that a second adherens junction protein, nectin is critical for tight junction assembly through binding the tight junction protein ZO-1, and disruption of tight junction assembly is independent of E-cadherin. α-catenin also binds ZO-1, but the consequences of this interaction are unknown. I hypothesized that α-catenin binding to ZO-1 plays a critical role in coupling tight junctions to adherens junctions. To test this, I mapped the ZO-1 binding site on α-catenin and engineered a point mutant of α-catenin that failed to bind ZO-1. Expression of this point mutant in epithelial cells showed that ZO-1 binding to α-catenin is essential for tight junction assembly and maintenance, while adherens junctions were unaffected. These findings established a role for ZO-1 binding to α-catenin in coupling tight junctions to adherens junctions during junction assembly, as well as at steady-state conditions. After discovering the importance of ZO-1 binding to α-catenin in coupling tight junctions to adherens junctions, I wanted to study whether this interaction is critical in a physiological setting. Tight junctions and adherens junctions are both strengthened in response to mechanical force; however the mechanisms responsible for tight junction strengthening were unknown. Using the system I previously developed, I show that ZO-1 binding to α-catenin is essential for increased tight junction integrity in response to mechanical force, coupling changes in tight junctions to increased stability of adherens junctions. Together, these findings identify a novel interaction that is critical for coupling tight junctions to adherens junctions under several conditions, and provide mechanistic insight into the cellular response to mechanical force.

Neisseria meningitidis ist mit jahrlich etwa 700.000 Erkrankungsfallen weltweit und einer Mortalitat von circa 7% einer der häufigsten Ausloser der bakteriellen Hirnhautentzündung. Der entscheidende Schritt zur Auslosung einer Meningitis ist die Uberwindung der Blut-Hirn-Schranke. Diese im menschlichen Korper einmalig dichte Barriere wird maßgeblich durch Tight-Junctions spezialisierter Endothelzellen der Hirnkapillaren aufrecht erhalten. Ob N. Meningitidis diese Barriere auf einem parazellulären oder transzellulärem Weg uberwindet, ist nicht vollstandig geklart. In dieser Arbeit wurde der Einfluss von N. meningitidis auf die Tight-Junction Proteine Occludin und ZO-1 unter Nutzung des HBMEC Zellkulurmodelles untersucht. Neben einer verminderten Genexpression von Occludin zeigte sich dabei eine Abspaltung eines 50 kDa Fragmentes von Occludin. Gleichzeitig konnte eine Umverteilung von Occludin von den Zellgrenzen in das Zytoplasma beobachtet werden. ZO-1 hingegen wurde weder in seiner Exprimierung, noch in seiner intrazellularen Verteilung beeinflusst. Mittels eines in dieser Arbeit etablierten Assays zur Bestimmung der Permeabilitat eines HBMEC-Monolayer als vereinfachtes in-vitro Modell der Blut-Hirn-Schranke konnte bestatigt werden, dass durch die Beeinflussung von Tight-Junction Proteinen die parazellulare Permeabilitat steigt. In weiteren Analysen konnten diese Prozesse auf eine gesteigerte Aktivitat von Matrixmetalloproteinase 8 zurückgefuhrt werden. Die Ergebnisse dieser Arbeit zeigen einen neuen Mechanismus auf, durch den N. meningitidis im Stande ist, die-Hinr-Schranke auf einem parazellulärem Weg zu überwinden. / Neisseria meningitidis is a leading cause of bacterial meningitis with about 700.000 cases per year and a mortality of approximately 7%. A crucial step in the pathogenesis of meningococcal meningitis is the crossing of the blood-brain barrier (BBB). This barrier mainly consists of tight junctions of the microvascular endothelial cells. It remains unclear whether N. meningitidis passes the BBB by a paracellular or transcellular route. In this work the influence of N. meningitidis on the tight junction proteins occludin and ZO-1 was investigated by using the HBMEC cell-culture model. Using QRT-PCR a reduced expression of occludin was seen, western-blot analyses showed a cleavage of occludin into a fragment of 50 kDA. Immonulourescence further showed a complete dissociation of occludin from the cell membrane, whereas ZO-1 was not influenced. By establishing a permeability-assay an increased permeability of the HBMEC monolayer after infection with N. meningitidis was demonstrated. Further work showed, that these effects are dependent of an increased activity of matrix metalloproteinase-8. These results reveal a new mechanism that could enable N. meningitidis to cross the BBB by a paracellular route.

Die Öffnung der Blut-Nerven-Schranke ist ein wichtiger Baustein in der Pathogenese neuropathischer Schmerzen. Die Blut-Nerven-Schranke schützt das periphere Nervensystem vor externen Einflüssen, wahrt die endoneurale Homöostase und trägt zur Aufrechterhaltung der neuronalen Signalweiterleitung bei. Sie wird durch die Pars epitheloidea des Perineuriums und endoneurale Gefäßzellen gebildet. Essentieller Bestandteil der Blut-Nerven-Schranke sind zwischen Perineural- und Gefäßzellen exprimierte Tight Junctions. Im Rahmen dieser Dissertation wurden die Tight Junction Proteine ZO-1, Claudin-1, -5, -19 und Occludin sowohl in einem Tiermodell neuropathischer Schmerzen, der Chronic Constriction Injury, als auch in Nervenbiopsien (N. suralis) von an Polyneuropathien erkrankten Patienten mittels Immunfluoreszenzfärbungen qualitativ und quantitativ untersucht. / The opening of the blood nerve barrier is an important in the pathogenesis of neuropathic pain. The blood nerve barrier protects the peripheral nervous system from external influences, preserves endoneurial homeostasis, and helps maintain neuronal signal transduction. It is formed by the pars epitheloidea of the perineurium and endoneural vascular cells. An essential component of the blood-nerve barrier are tight junctions expressed between perineural and vascular cells. In this thesis, the tight junction poteins ZO-1, claudin-1, -5, -19 and occludin were examined qualitatively and quantitatively in an animal model of neuropathic pain, the Chronic Constriction Injury, as well as in human nerve biopsies (N. suralis) by patients suffering from polyneuropathy using immunofluorescence stainings.

The Sertoli cell tight junction (TJ) of the seminiferous epithelium is important for the developmental process of spermatogenesis as it separates germ cells in the seminiferous tubules from the general circulation in the testicular interstitium. Absence of the TJ leads to spermatogenic arrest and infertility. TJs form at puberty as circulating gonadotrophins luteinising hormone/testosterone and follicle stimulating hormone increase. Several studies have demonstrated hormonal regulation of the two major TJ proteins, claudin-11 and occludin, and also of TJ function in vitro and in vivo. Men with low levels of circulating gonadotrophins exhibit an immature and dysfunctional TJ phenotype, which is reversed upon the exogenous application of gonadotrophins. This thesis hypothesises that claudin-11 and occludin are the major contributors to TJ function, and that gonadotrophins regulate TJ function and structure via these two proteins in several species including humans. This PhD was divided into four separate studies to address these hypotheses. The first study selectively silenced the genetic expression of claudin-11 and occludin with small interfering RNA (siRNA) in cultured immature rat Sertoli cells to determine their contribution to Sertoli cell TJ function in vitro. siRNA treatment against either protein significantly (p less than 0.01) reduced TJ function by ~50% as assessed by transepithelial electrical resistance. Immunocytochemistry displayed marked reductions in the localisation of these proteins to the TJ after siRNA treatment. It was concluded that both proteins significantly contributed to TJ function in vitro. The second and third studies then aimed to study hormonal regulation of the TJ in vivo. Weekly injections of the gonadotrophin releasing hormone antagonist acyline were used to suppress circulating gonadotrophins and spermatogenesis in adult rats. Acyline treatment disrupted i) the localisation of occludin to the TJ and ii) TJ function as shown by permeability to a biotin tracer, which was impermeable to TJs in controls. Short-term hormone replacement partially restored the effects of gonadotrophin suppression. It was concluded that gonadotrophins regulate the maintenance of the TJ in rats in vivo. The third study used the hypogonadal (hpg) mouse, which is a naturally occurring model of gonadotrophin deficiency with inactive spermatogenesis. Claudin-11 in hpg mice was not localised at the TJs, and these were dysfunctional as shown by permeability to biotin. Following hormone treatment, TJs were structurally and functionally competent, demonstrating that gonadotrophins also regulate the formation of TJs in vivo. The fourth study subsequently analysed TJs in gonadotrophin suppressed men, and it was found that claudin-11 staining was reduced from continuous bands in control men, to punctate staining in gonadotrophin-suppressed men, demonstrating that gonadotrophins also regulate the localisation of claudin-11 to the TJ in men in vivo. In summary, it is concluded that the Sertoli cell TJ is hormonally regulated, and that the major contributors to TJ function in vivo and in vitro are claudin-11 and occludin. It is hypothesised that the reduction of claudin-11 localisation to the TJ in men may also result in a loss of human Sertoli cell TJ function, suggesting that the TJ may be a potential target of hormonal contraception in men.

<p>In phylogenetic analysis one study the relationship between different species. By comparing DNA from two different species it is possible to get a numerical value representing the difference between the species. For a set of species, all pair-wise comparisons result in a dissimilarity matrix <em>d</em>.</p><p>In this thesis I present a few methods for constructing a phylogenetic tree from <em>d</em>. The common denominator for these methods is that they do not generate a tree, but instead give a connected graph. The resulting graph will be a tree, in areas where the data perfectly matches a tree. When <em>d</em> does not perfectly match a tree, the resulting graph will instead show the different possible topologies, and how strong support they have from the data.</p><p>Finally I have tested the methods both on real measured data and constructed test cases.</p>

In phylogenetic analysis one study the relationship between different species. By comparing DNA from two different species it is possible to get a numerical value representing the difference between the species. For a set of species, all pair-wise comparisons result in a dissimilarity matrix d. In this thesis I present a few methods for constructing a phylogenetic tree from d. The common denominator for these methods is that they do not generate a tree, but instead give a connected graph. The resulting graph will be a tree, in areas where the data perfectly matches a tree. When d does not perfectly match a tree, the resulting graph will instead show the different possible topologies, and how strong support they have from the data. Finally I have tested the methods both on real measured data and constructed test cases.

Time-rate analysis and time-rate-pressure analysis methods are available to estimate reserves and study flow performance of wells in unconventional gas reservoirs. However, these tools are often incorrectly used or the analysis can become difficult because of the complex nature of the reservoir system. Conventional methods (e.g., Arps' time-rate relations) are often used incorrectly to estimate reserves from such reservoirs. It was only recently that a serious study was conducted to outline the limitations of these relations and to set guidelines for their correct application. New time-rate relations, particularly the Duong and logistic growth model, were introduced to estimate reserves and forecast production from unconventional reservoirs. These new models are being used with limited understanding of their characteristics and limitations. Moreover, well performance analyses using analytical/semi-analytical solutions (time-rate-pressure) are often complicated from non-uniqueness that arises when estimating well/formation properties. In this work, we present a detailed study of the Duong model and logistic growth model to investigate the behaviors and limitations of these models when analyzing production data from unconventional reservoirs. We consider production data generated from numerical simulation cases and data obtained from unconventional gas reservoirs to study the quality of match to specific flow regimes and compare accuracy of the reserve estimates. We use the power-law exponential model (PLE), which has been shown to model transient, transition and boundary-dominated flow regimes reliably, as a benchmark to study performance of Duong and logistic growth models. Moreover, we use the "continuous EUR" approach to compare these models during reserve estimation. Finally, we develop four new time-rate relations, based on characteristics of the time-rate data on diagnostic plots. Using diagnostic plots we show that the new time-rate relations provide a quality match to the production data across all flow regimes, leading to a reliable reserve estimate. In a preliminary study, we integrated time-rate model parameters with fundamental reservoir properties (i.e., fracture conductivity (Fc) and 30 year EUR (EUR30yr)), by studying 15 numerical simulation cases to yield parametric correlations. We have demonstrated a methodology to integrate time-rate model parameters and reservoir properties. This method avoids the non-uniqueness issues often associated with model-based production data analysis. This study provides theoretical basis for further demonstration of the methodology using field cases.

This thesis study is about the effects of tight budgetary control on managerial behaviour in the Swedish public sector. Managerial behaviour represents the four variables; motivation, organizational commitment, stress, and satisfaction. As previous research mostly investigated the effects of tight budgetary control on organizational performance in the private sector, this study can be seen as one of the first that focus on managerial behaviour and is based on the public sector. For the data collection process, an online questionnaire was created that has been distributed via email to the different participants. Moreover, the study is based on a sample of 62 managers from different municipalities and universities throughout Sweden. The result of the thesis research study revealed that the majority of the local managers in the Swedish public sector actually experience tight budgetary control. Furthermore, the statistical test of the created hypotheses showed that the behavioural factors, both, organizational commitment and stress were respectively negatively and positively related with tight budgetary control. However, for the behavioural variables of motivation and satisfaction no significant results have been found.

It is known that a highly ordered medium allows certain wave functions to move unhindered throughout and in this manner achieve delocalization. It is also known that if one introduces disorder into a medium, wave packets will not be able to move as freely and will instead be trapped or localized. In this thesis, I have simulated a medium in which the amount of disorder can be modified and using this I have shown that the shape of the localization can be altered.