Accurate and efficient genome editing is primarily dependent on the generation of a sequence-specific, genomic double-stranded DNA break (DSB) combined with the introduction of an exogenous DNA... Show moreAccurate and efficient genome editing is primarily dependent on the generation of a sequence-specific, genomic double-stranded DNA break (DSB) combined with the introduction of an exogenous DNA template into target cells. The exogenous template, called donor DNA, normally contains the foreign sequences flanked by DNA regions sharing sequence identity ("homologous") to those bracketing the target site. The strategies for mediating the formation of DSBs at the predefined genomic loci, have been undergoing intense investigation since the introduction of sequence-customizable zinc-finger nuclease (ZFN) technology. More recently, prokaryotic protein-based transcription activator-like effector nucleases (TALENs) and RNA-guided nucleases (RGNs) derived from CRISPR-associated protein (Cas9) complexes have substantially broadened the availability and applicability of designer nuclease-mediated genome editing. A potential alternative research line to the use of designer nucleases, is to investigate whether specific DNA structures can, by themselves, serve as triggers of the DNA damage response and, in doing so, elicit targeted gene repair. Such an approach would simplify genome editing protocols, such as, by reducing the number of reagents needed to be introduced into target cells. In this thesis, the roles of these secondary structures as well as designer nucleases and donor-DNA templates, delivered via adenoviral vectors, is described. Show less
Evaluation of patients with ischemic heart disease using echocardiography is indispensable. Both in the acute setting of STEMI, at follow-up and in the chronic phase during the possible development... Show moreEvaluation of patients with ischemic heart disease using echocardiography is indispensable. Both in the acute setting of STEMI, at follow-up and in the chronic phase during the possible development of heart failure, routine echocardiographic assessment is an essential part of daily clinical practice. Show less
This thesis discusses the negotiation of custom in the Landraad, a judicial forum set up by the Dutch East India Company (VOC), in eighteenth-century Sri Lanka. The Landraad had a majority of VOC... Show moreThis thesis discusses the negotiation of custom in the Landraad, a judicial forum set up by the Dutch East India Company (VOC), in eighteenth-century Sri Lanka. The Landraad had a majority of VOC officials and a subordinate minority of local headmen. It was a space in which agency was multifarious. The nuances of everyday practice as studied here reveal both rejection and manipulation by local actors. This in turn informs us of everyday life in early-modern colonialism. A choice of laws came into play, that choice being significant at varying degrees for different areas of the law such as evidence, inheritance, land and marriage law. Based on over a hundred cases brought forward by men and women from the Galle district in southern Sri Lanka, the lived experience of legal pluralism is emphasised. This thesis adds an empirical study and the insights of socio-legal studies of legal pluralism to the existing literature on Roman-Dutch law in Sri Lanka. While there was inevitable conflict, in practice the local normative order was as much a social fact for the early colonial rulers as Roman-Dutch law. Show less
The research in this thesis was aimed at identifying and understanding mechanisms underlying modulating factors for and consequences of cortical spreading depression (CSD), the pathophysiological... Show moreThe research in this thesis was aimed at identifying and understanding mechanisms underlying modulating factors for and consequences of cortical spreading depression (CSD), the pathophysiological substrate for migraine aura that occurs in one-third of migraine patients. In this thesis, experimental studies on CSD were performed in wild-type (WT) and transgenic migraine mice, which express CaV2.1 Ca2+ channels with a mutated _1 subunit that contains the R192Q missense mutation. The R192Q mutation was previously identified in patients with familial hemiplegic migraine type 1 (FHM1) and causes gain-of-function effects in terms of neuronal Ca2+ influx, neurotransmission, and susceptibility to experimentally induced CSD. Using various experimental strategies, in this thesis, the FHM1 R192Q mouse model was used to study pathophysiological mechanisms of the initiation and modulation of CSD as well as of neurobiological and molecular changes that accompany CSD events. Show less
Persistent infections with high-risk type human papillomaviruses (hrHPVs) can progress to cancer. HrHPVs infect keratinocytes (KCs) and successfully suppress host immunity for up to two years... Show morePersistent infections with high-risk type human papillomaviruses (hrHPVs) can progress to cancer. HrHPVs infect keratinocytes (KCs) and successfully suppress host immunity for up to two years despite the fact that KCs are well equipped to detect and initiate immune responses to invading pathogens. HrHPV interferes with the innate immune response by affecting several signaling pathways that otherwise would prompt anti-viral mechanisms in the host cell. Furthermore, hrHPV interferes with the production of cytokines that are involved in the attraction of immune cells to the infected epithelium. In addition, hrHPV hides itself from the immune system by suppressing the antigen presentation machinery and employs means to hamper the response of KC__s to signals from adaptive immune cells. In this thesis we show that hrHPV attenuates innate immune signaling (Chapter 2) and CD40-mediated (Chapter 3) and IFN_ and/or TNF_-induced (Chapter 4) adaptive immune signaling. HrHPV exploits the cellular proteins UCHL1 (Chapter 2) and IFRD1 (Chapter 4) that act on multiple points in the IRF and NF_B signaling pathways. Moreover, hrHPV downregulates cellular IFITM1 to resist the growth inhibitory effects of IFN_ and/or TNF_ (Chapter 5). Our data provide important new insights on how hrHPV can persist in the face of host immunity. Show less
In this thesis the epidemiology of suicidal ideation and suicide attempts (together referred to as 'suicidality') in Huntington's disease (HD) is investigated and coping styles and support... Show moreIn this thesis the epidemiology of suicidal ideation and suicide attempts (together referred to as 'suicidality') in Huntington's disease (HD) is investigated and coping styles and support strategies that may serve to help suicidal HD mutation carriers are explored. Suicidality frequently occurs in HD, with a 1-month prevalence of up to 20%. Mutation carriers who were most likely to currently experience suicidal ideation or suicidality had a shorter disease duration, were anxious, aggressive, previously attempted suicide, used antidepressants, and had a depressed mood. The presence of a depressed mood and use of benzodiazepines were the only significant independent predictors of incident suicidal ideation or suicidality. Biological parameters, in particular functioning of the immune system and the hypothalamus-pituitary-adrenal axis were not found to be associated with suicidality in HD. In a qualitative study it was shown that suicidal HD mutation carriers generally used four strategies to cope with suicidality: talking about suicidality, employing self-management activities, using medication, and discussing end-of-life wishes. Lastly, in a meta-analysis, we examined whether the expression of suicidal ideation predicted subsequent completed suicide. Effect estimates differed substantially among different populations, but none of the included studies investigated this association in HD. Show less
“Transitional justice” is a field of practice, policy and study that focuses on the ways that societies respond to legacies of large-scale atrocities though tribunals, truth commissions,... Show more“Transitional justice” is a field of practice, policy and study that focuses on the ways that societies respond to legacies of large-scale atrocities though tribunals, truth commissions, reparations, and other mechanisms. Over the last thirty years, transitional justice has become the globally dominant lens through which we grapple with such legacies. Yet transitional justice practice has rarely reflected the diversity of peace and justice traditions around the world, bound up as it has been with largely liberal and Western conceptions of justice. This thesis examines the frictions and blindspots these dynamics have created, arguing that the liberal transitional justice lens has significantly biased our conceptions of what it means to “do justice” as well as the modalities for bringing justice about. If we are to make transitional justice into a true global project, I argue, we need to revisit and deconstruct the field’s core normative metanarratives and assumptions as a prelude to seeking a more emancipatory ground for transitional justice policy and practice that is true to human rights ideals while becoming more open-textured and attuned to local needs and context. Show less
Asthma is a common non-communicable respiratory disease. In this thesis we analysed three different management strategies for adult patients with asthma in primary care. In the first, we targeted... Show moreAsthma is a common non-communicable respiratory disease. In this thesis we analysed three different management strategies for adult patients with asthma in primary care. In the first, we targeted the currently recommended aim of __Controlled asthma__, which means patients experience hardly any symptoms. The second targeted __Partly Controlled asthma__, which allowed some symptoms. The third additionally used a FeNO-measurement to assess current control. Aiming for Partly Controlled asthma significantly reduced medication usage compared to Controlled asthma, while exacerbation-rate, quality of life and asthma control were similar. FeNO-guided Controlled asthma also significantly reduced medication usage compared to Controlled asthma and additionally significantly improved asthma control compared to Partly Controlled asthma. Aiming for FeNO-guided Controlled asthma is therefore the optimal strategy. In this thesis we also analysed daily measurements of symptoms and peak-flow, which are recorded in Written Asthma Action Plans, or in apps. Hereby we were able to predict future asthma exacerbations several days in advance with reasonable accuracy. Finally, we compared an online assessment of asthma control by a questionnaire with the results of the same questionnaire assessed by a practice nurse. We showed that asthma control is perceived as better when interacting with a caregiver than by online self-assessment. Show less
Electroconvulsive therapy (ECT) is the transcutaneous application of small electrical stimuli to the brain to produce generalized seizure for the treatment of selected psychiatric disorders, mostly... Show moreElectroconvulsive therapy (ECT) is the transcutaneous application of small electrical stimuli to the brain to produce generalized seizure for the treatment of selected psychiatric disorders, mostly treatment resistant depression, acute mania, and schizophrenic syndromes. Safety of ECT increases the efficacy of therapy and provides fulfillment of a series of required treatments resulting in longer treatment effect of ECT. During the last few decades, researchers have been attempting to improve the effectiveness of ECT, to learn how and why it works, and to understand its risks and adverse side effects. As a result, the safety and efficacy of ECT has been improved and its indications have been relatively defined to increase the efficiency of outcome of therapy. While there has been considerable improvement in safety features, further investigation have been done to promote both the safety and efficacy of the treatment. Such efforts could increase our knowledge on the biological mechanisms involved in effectiveness of ECT that might result in discovery of new treatments. In this dissertation, we investigate how preprocedural medications could improve the safety and efficacy of ECT and further investigated the potential role of pharmacogenetics in the efficacy of ECT and procedural side effects such as cognitive disorders. Show less
Super-Resolution Microscopy is an optical fluorescence technique. In this thesis we focus on single molecule super-resolution, where the position of single molecules is determined. Typically these... Show moreSuper-Resolution Microscopy is an optical fluorescence technique. In this thesis we focus on single molecule super-resolution, where the position of single molecules is determined. Typically these molecules can be localized with a 10 to 30nm precision. This technique is applied in four different studies. To determine the spatial distribution of Ras-protein in live cells, Ripley's analysis is used on localization data to quantify size an diffusion parameters of nanodomains. Particle image correlation spectroscopy (PICS) is a second order spatial distribution analysis to determine diffusion properties of single molecule populations. A mathematical framework to correct the data when applied on 3D diffusion is presented in the second chapter. In the fourth chapter the uptake of alpha-synuclein aggregates by the cells is observed using super-resolution microscopy. Their partial degradation was followed and showed the importance of the lysosome-dependent mechanism for protecting cells from exposure to potentially toxic a-synuclein. In the fifth chapter we correlate the number of vinculin proteins in a focal adhesion protein complex, to the local force generated by the cell via this complex. A method was developed to determine the local stoichiometry of molecules by their correlated distances as obtained from SMLM. Show less
In the Netherlands, levying tax on inheritances at death has been a fact of life for centuries. The presumed double taxation and its sad precursor are elements of the discussion about the levy of... Show moreIn the Netherlands, levying tax on inheritances at death has been a fact of life for centuries. The presumed double taxation and its sad precursor are elements of the discussion about the levy of this inheritance tax, but so too are the possibilities to reduce the tax through ingenious legal constructions. In its zeal to counter such clever plans the legislature has turned the law into a complex body without parrying all the avoidance behaviour. On top of that, the effects of the current law can be very unreasonable. The thesis uncovers the root cause of this problem by retracing the levy of inheritance tax to the Republic of the Seven United Netherlands, a little-described chapter in legal history. It is discovered how the current problems would never have occurred under the simple statutory provision applicable in that period. The obvious conclusion is that the inheritance tax should be reformed based on that model. How such amendment can best be designed is elaborated further based on insights gained from legal history and other legal systems. This has resulted in a complete bill with comments, so the legislature now has the solution at its fingertips. Show less
Minor histocompatibility antigens (MiHA) are structures that can be recognized by T cells. In the setting of allogeneic stem cell transplantation MiHA presented by patient cells can be recognized... Show moreMinor histocompatibility antigens (MiHA) are structures that can be recognized by T cells. In the setting of allogeneic stem cell transplantation MiHA presented by patient cells can be recognized by donor T cells. If MiHA that are targeted by donor T cells are selectively expressed by leukemic cells, beneficial graft-versus-leukemia reactivity can be observed. However, if MiHA are presented by normal tissue cells, graft-versus-host disease can be a serious complication. The thesis describes methods that were applied to characterize the molecular structure of MiHA. T cells that recognized MiHA recognized on malignant hematopoietic and solid tumor cells were identified. MiHA recognition on both normal and malignant cells was measured. We finally investigated the differences of the donor T cell response of patients with a clinical graft-versus-leukemia reactivity who did or did not develop graft-versus-host disease. Show less
The concept of topology, having played a perpetual center stage role in the field of mathematics and physics for already a few centuries, heuristically pertains to the study of properties of... Show moreThe concept of topology, having played a perpetual center stage role in the field of mathematics and physics for already a few centuries, heuristically pertains to the study of properties of geometrical objects that are preserved under smooth deformations. Recently, it was uncovered that such topological structures may in particular emerge in the quantum world of electron systems that are subjected to the presence of discrete symmetries such as time reversal symmetry, particle hole symmetry and chiral symmetry. Nonetheless, the role of the most prominent and necessarily present symmetry in this regard, being that of the underlying lattice, has remained elusive. In this work we signify the profound interplay between topological order in electronic band theory and the associated crystal symmetries by exploring the physics of defects, with the emphasis on dislocations. We find that in addition to a plethora of interesting mechanisms, which are related to the defects themselves and culminate, inter alia, in helical propagating fermionic modes bound to freely deformable channels and exotic isospinless graphene-like states that can be linked to illustrious field theoretical anomalies, these results in turn expose an extension of the classification of topological electrons systems. Show less
This thesis focused on the development of creative thinking across adolescence and into adulthood. To this end, a range of creativity tasks, both with and without an fMRI scanner, and before and... Show moreThis thesis focused on the development of creative thinking across adolescence and into adulthood. To this end, a range of creativity tasks, both with and without an fMRI scanner, and before and after training paradigms has been used to ex_amine both age- and experience-related effects on creative thinking performance during functional brain develop_ment. Chapter 1 provides a theoretical background for the research described in chapters 2 to 6. In Chapter 2, developmental trajectories of creative cognition across adolescence and early adulthood were examined using a set of tasks gauging both insight and divergent thinking in the verbal and visual domain. In Chapter 3, behavioral and neural differences for creative problem solving in middle-adolescents and adults were examined. Chapter 4 examined the neural correlates of divergent thinking in adults and adolescents. Chapter 5 focused on the effectiveness of creative ideation training in adolescents and adults. In Chapter 6, the benefits of training creativity in adolescents were examined using fMRI techniques. Finally, Chapter 7 summarizes the main results of the empirical studies presented in this thesis. Here, implications of the results are discussed and suggestions for future research are presented. Show less
Rheumatoid arthritis (RA) is a chronic, systemic autoimmune disease that principally affects synovial joints. RA is present in 0,5% to 1% of the global population. The incidence of RA is higher in... Show moreRheumatoid arthritis (RA) is a chronic, systemic autoimmune disease that principally affects synovial joints. RA is present in 0,5% to 1% of the global population. The incidence of RA is higher in women than in men and increases with age (1). RA can affect any joint but preferably small joints in hands and feet (2). The symptoms of RA include pain, swelling, stiffness, redness, warmth and can finally lead to loss of joint functions (2). The systemic symptoms of RA include fatigue, malaise, loss of appetite and muscle ache. Next to the joints RA can affect other organs such as skin, lungs, heart and blood vessels (3). RA can be classified using the 2010 American College of Rheumatology/European League Against Rheumatism classification criteria for rheumatoid arthritis (2010 ACR/EULAR criteria) (4). This is a quantitative system in which scores can be obtained from: joint involvement, serologic markers, inflammation markers and duration of symptoms (4). Show less
