BackgroundMedical device certification has undergone significant changes in recent years. However, exploration of stakeholder experiences remains relatively limited, particularly in the context of... Show moreBackgroundMedical device certification has undergone significant changes in recent years. However, exploration of stakeholder experiences remains relatively limited, particularly in the context of software as a medical device. This study sought to explore stakeholder experiences of medical device certification across both the UK and EU.MethodsSemi-structured interviews (n = 22) analysed using inductive-thematic analysis, synthesised using activity theory.ResultsInnovators, consultants and notified bodies share more similarities than differences when discussing barriers and enablers to achieving medical device certification. Systemic tensions between existing rules, tools, community understanding and division of labour currently undermine the intended aim of certification processes. Existing rules are considered complex, with small and medium-sized enterprises considered disproportionality affected, resulting in several unintended outcomes including the perceived ‘killing’ of innovation. Existing certification processes are described as unfit for purpose, unethical and unsustainable.ConclusionStakeholder experiences suggest that the intention of establishing a robust and sustainable regulatory framework capable of ensuring a high level of safety whilst also supporting innovation is not yet being realised. Failure to enact desired changes may further jeopardise future innovations, outcomes and care quality. Show less
Daskalakis, N.P.; Meijer, O.C.; Kloet, E.R. de 2022
'You can't roll the clock back and reverse the effects of experiences' Bruce McEwen used to say when explaining how allostasis labels the adaptive process. Here we will for once roll the clock back... Show more'You can't roll the clock back and reverse the effects of experiences' Bruce McEwen used to say when explaining how allostasis labels the adaptive process. Here we will for once roll the clock back to the times that the science of the glucocorticoid hormone was honored with a Nobel prize and highlight the discovery of their receptors in the hippocampus as inroad to its current status as master regulator in control of stress coping and adaptation. Glucocorticoids operate in concert with numerous neurotransmitters, neuropeptides, and other hormones with the aim to facilitate processing of information in the neurocircuitry of stress, from anticipation and perception of a novel experience to behavioral adaptation and memory storage. This action, exerted by the glucocorticoids, is guided by two complementary receptor systems, mineralocorticoid receptors (MR) and glucocorticoid receptors (GR), that need to be balanced for a healthy stress response pattern. Here we discuss the cellular, neuroendocrine, and behavioral studies underlying the MR:GR balance concept, highlight the relevance of hypothalamic pituitary-adrenal (HPA)-axis patterns and note the limited understanding yet of sexual dimorphism in glucocorticoid actions. We conclude with the prospect that (i) genetically and epigenetically regulated receptor variants dictate cell-type-specific transcriptome signatures of stress-related neuropsychiatric symptoms and (ii) selective receptor modulators are becoming available for more targeted treatment. These two new developments to 'restart the clock' with the to resilience. Show less
Bacteria are highly complex and diverse organisms that have adapted to survive in ecological niches ranging from the most extreme to the most heterogeneous environments. Actinobacteria, with their... Show moreBacteria are highly complex and diverse organisms that have adapted to survive in ecological niches ranging from the most extreme to the most heterogeneous environments. Actinobacteria, with their beautiful morphologies and complex multicellular life cycle, are a striking example. These filamentous bacteria produce many enzymes and natural products, including two-thirds of all known antibiotics, which makes them highly relevant for medical, biotechnological and industrial applications. In Streptomyces, complex and intertwining regulatory networks, consisting of pathway-specific and global regulators, control development and antibiotic production in response to environmental stimuli and stresses. This thesis explores how the model organism Streptomyces coelicolor senses environmental signals, related to oxygen stress and the availability of aminosugars, and how the bacterium translates these into appropriate responses. Show less
Vermindering van regeldruk maakt al decennialang deel uit van het Nederlandse overheidsbeleid. Zowel op rijksniveau als op lokaal niveau hebben verschillende programma’s de revue gepasseerd om... Show moreVermindering van regeldruk maakt al decennialang deel uit van het Nederlandse overheidsbeleid. Zowel op rijksniveau als op lokaal niveau hebben verschillende programma’s de revue gepasseerd om het aantal regels terug te brengen, regels efficiënter te maken en het toezicht beter af te stemmen. De voortgangsrapportages van de overheid zeggen dat de aanpak succesvol is: deze zou het bedrijfsleven reeds miljarden aan besparingen hebben opgeleverd. Ondernemers zeggen daar echter weinig van te merken. Ook de Algemene Rekenkamer constateerde in 2006 dat de successen in de samenleving niet als zodanig voelbaar waren. In deze studie onderzoekt Esmeralda Vergeer het fenomeen regeldruk. Ze brengt in beeld hoe de regeldrukaanpak van de Nederlandse overheid zich in de loop der jaren ontwikkelde en hoe er in de wetenschap naar regeldruk wordt gekeken. Verschillende definities van regeldruk passeren de revue, maar geen ervan blijkt onomstreden. Vergeer richt zich op ondernemers en constateert dat zij een eigen, geheel nieuwe definitie toekennen aan regeldruk. Ze onderscheidt drie factoren die van invloed zijn op de ervaring van regeldruk: kosten, werkbaarheid en gepercipieerd nut. Het onderzoek biedt zo nieuwe inzichten om regeldruk voor ondernemers merkbaar te verminderen. Show less
The research considers structures and practices of regulatory inspections and enforcement, and particularly the relevance and effects of purported “smarter inspections” methods, e.g. risk... Show more The research considers structures and practices of regulatory inspections and enforcement, and particularly the relevance and effects of purported “smarter inspections” methods, e.g. risk-based ones, and how they compare with previously existing practices. Specifically, it considers efficiency and effectiveness of inspections in terms of achieving public welfare, and their economic impact. The first part considers the history of regulatory inspections and how specific inspection institutions started to appear from the 19th century onwards. Of particular interest is how path dependency resulted in important differences in structures and methods between different fields and countries, and how these in turn produced different effects for the public. The second part looks at existing research on the interconnected issues of regulatory compliance drivers, regulatory discretion and risk and regulation. It concludes to the importance of combining several compliance drivers to achieve better results, to the appropriateness of structuring discretion in inspections and enforcement, and to the adequacy of “risk” as an instrument to do so. The third part looks at several case studies (in particular occupational safety and health in Britain and Germany) to consider whether risk-based, ‘smart’ approaches effectively deliver better results with lower burden, and tentatively concludes to the affirmative. Show less
In this thesis, we have addressed aspects of two main arms of the adaptive immune system; the B cell and antibody arm and the T cell arm. This led to a division in the presentation of the... Show more In this thesis, we have addressed aspects of two main arms of the adaptive immune system; the B cell and antibody arm and the T cell arm. This led to a division in the presentation of the results described in this thesis into two sections. In the first section, we present the results regarding the characterization of ACPA responses, B cells and ACPA secreting plasmablasts/-cells in RA as well as autoantibody responses and their regulation by an effective anti-rheumatic drug, abatacept, in the arthritis mouse model; Collagen Induced Arthritis (CIA). The second section is compiled of results obtained from studies examining the regulatory and other aspects of CD49b+CD4+ T cells on proinflammatory responses involved in the pathogenesis of arthritis. Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by inflammation of the synovial membrane of the joints, culminating in destruction of cartilage and deformity of the joints if remains untreated. Infiltration of inflammatory immune cells such as B cells and T cells into the inflamed joints is a characteristic feature of RA. These immune cells are in continuous interaction with each other and create a viscous circle that sustains persistent synovitis and damage to articular cartilage. Show less
Belo, N.A.H.; Driel, J.H. van; Veen, K. van; Verloop, N. 2014
Members of the order Nidovirales (Coronaviridae, Arteriviridae, and Roniviridae) employ genomes with mRNA polarity (plus-strand) and encode one of the most complex RNA virus replicases currently... Show moreMembers of the order Nidovirales (Coronaviridae, Arteriviridae, and Roniviridae) employ genomes with mRNA polarity (plus-strand) and encode one of the most complex RNA virus replicases currently known. This replicase is expressed from the viral genome by translation of two large 5__-proximal ORFs into two polyproteins, which are processed by virus-encoded proteases in 13-16 individual nonstructural proteins (nsps). The nsps direct the formation of an RNA-synthesizing complex that mediates viral genome replication, as well as the synthesis of a 3__-coterminal nested set of subgenomic (sg) mRNAs, from which the viral genes located downstream of the replicase gene are expressed. Arteriviruses and coronaviruses presumably employ a unique mechanism of discontinuous minus-strand extension to generate subgenome-length templates for sg mRNA synthesis. This thesis focused on the functional characterization of two replicase subunits and their roles in coupling different processes in the replicative cycle of equine arteritis virus (EAV), the arterivirus prototype. The biological importance of a conserved domain unique to nidoviruses (NendoU), mapping to arterivirus nsp11 and proposed to function as an endoribonuclease, was addressed. We demonstrated the recombinant nsp11 exhibits pyrimidine-specific endoribonuclease activity, and showed the critical importance of the NendoU domain for EAV RNA synthesis. In addition, we identified a multidomain replicase subunit, EAV nsp1, as a key coordinator of EAV genome replication, sg mRNA synthesis, and virus production. Our results reveal that the relative abundance of EAV mRNAs is tightly controlled by nsp1 and is critical for efficient production of new virus particles. The protein was implicated in modulating the accumulation of full-length and subgenome-length minus-strand templates for EAV mRNA synthesis. A protocol for purification of soluble recombinant nsp1, which can be used in future research on the molecular mechanisms of nsp1 function, is described. Show less
The topic of this thesis is the molecular regulation of Hypoxia-Inducible Factors (HIFs) by sumoylation. Hypoxia-Inducible Factors are transcriptional regulators that mediate cell survival during... Show moreThe topic of this thesis is the molecular regulation of Hypoxia-Inducible Factors (HIFs) by sumoylation. Hypoxia-Inducible Factors are transcriptional regulators that mediate cell survival during reduced oxygen conditions (hypoxia). HIFs are extensively controlled by post-translational modifications. The aim of this thesis was to obtain novel insight in the posttranslational modification of HIFs by sumoylation. Chapters 1 and 2 are introductions on sumoylation and Hypoxia-Inducible Factors, respectively. A summary of current knowledge is provided based on published literature on both systems. Chapter 3 describes the regulation of HIF-1_ transcriptional activity by sumoylation via a novel SUMO acceptor site. Chapter 4 describes crosstalk between sumoylation and ubiquitination, two similar but distinct post-translational modifications that act in concert to regulate the proteasomal degradation of a subset of protein targets. Chapter 5 shows that sumoylation of HIF-2_ marks it for proteasomal degradation during hypoxia, facilitated by E3 ubiquitin ligases RNF4 and VHL. Chapter 6 presents data on the formation of SUMO chains made up of a mix of SUMO-1 and SUMO-2/3 and we show that HIF-1_ is a target for SUMO chains in vitro. Chapter 7 provides a summary and overall discussion of the thesis. Show less
he creation of European Union agencies is arguably one of the most prominent institutional innovations at the EU level in recent history. Especially since the early 1990s, the EU and its member... Show morehe creation of European Union agencies is arguably one of the most prominent institutional innovations at the EU level in recent history. Especially since the early 1990s, the EU and its member states delegated a wide range of (semi-)regulatory, monitoring, and coordination tasks to a quickly growing number of agencies. Most existing research focuses on the creation of these agencies. As a result, we do not know much about how agencies develop after their creation. EU agencies are formally independent, but do they also behave autonomously in practice? How does actual autonomy vary across EU agencies and how does this affect the role these agencies play in the multi-level system of European governance? This study addresses these questions theoretically and empirically by comparing six EU agencies – the European Medicines Agency (EMEA), the European Food Safety Authority (EFSA), the European Environm ent Agency (EEA), the European Monitoring Centre for Racism and Xenophobia (EUMC), Europol and Eurojust. It shows how some of these agencies develop into relatively autonomous entities by acquiring a distinct organisational character and by generating support from actors in their environment, whereas other agencies do so to a much lesser extent or not at all. Show less
The European natural gas market is in a state of flux. In order to better secure the public service obligations supply security, competitiveness and sustainability this market is currently being... Show moreThe European natural gas market is in a state of flux. In order to better secure the public service obligations supply security, competitiveness and sustainability this market is currently being liberalized. This requires significant structural and regulatory reform. This reform process is not yet finished; the European gas market finds itself in a transition phase from the traditional managed market structure towards an internal European gas market which is governed by competitive forces. In such a transition phase, problems and tensions are inevitable. This study assesses the adequacy of the current approach towards structural and regulatory reform of the European gas market in light of the changes that are taking place on this market. These changes create a new context for gas regulation. Given the vital role of investments in securing any of the public service obligations, the analysis is conducted from the perspective of ensuring sufficient investments. The theoretical part criticizes the current neoclassical approach towards reform and argues that the transaction cost perspective will result in better outcomes in terms of market behavior in the new regulatory context. The empirical part undertakes case studies on two recent amendments to gas regulation the possibility for certain investments to be exempted from the provisions to provide third party access, and the enlarged scope for concluding downstream long-term gas supply contracts in order to assess whether the theoretical critique is also valid in practice. Both case studies confirm the theoretical critique. The current regulatory approach appears to be stuck between the neoclassical and the transaction cost perspective. Recommendations are provided in order to better align current regulation with the insights provided by Transaction Cost Economics. Show less
Plants produce a variety of secondary metabolites. In Catharanthus roseus, several have pharmaceutical applications, including the monomeric alkaloids serpentine and ajmalicine, which are used as a... Show morePlants produce a variety of secondary metabolites. In Catharanthus roseus, several have pharmaceutical applications, including the monomeric alkaloids serpentine and ajmalicine, which are used as a tranquillizer and to reduce hypertension, respectively, and the dimeric alkaloids vincristine and vinblastine, which are potent antitumour drugs. Jasmonic acid (JA) is a key defense hormone, which controls the expression of several alkaloid biosynthesis genes in Catharanthus. The JA-responsive expression is controlled by the AP2-domain transcription factors ORCA2 and ORCA3. The results in this thesis demonstrate that JA-responsive ORCA expression is controlled by an up-stream positive regulator, whose activities are affected by a family of repressors. Understanding the regulation mechanism may help to generate a very valuable tool for engineering the production of valuable secondary metabolites. Show less
Streptomycetes are Gram-positive soil-dwelling bacteria, in appearance similar to filamentous fungi. The SsgA-like proteins or SALPs, of which streptomycetes typically have at least five paralogues... Show moreStreptomycetes are Gram-positive soil-dwelling bacteria, in appearance similar to filamentous fungi. The SsgA-like proteins or SALPs, of which streptomycetes typically have at least five paralogues, control specific steps of sporulation-specific cell division in streptomycetes. The expression level of SsgA, the best studied SALP, has a rather dramatic effect on septation and on hyphal morphology, which is not only of relevance for our understanding of (developmental) cell division but has also been succesfully applied in industrial fermentation, to improve growth and production of filamentous actinomycetes. The different regulation of ssgA transcription in different Streptomyces species, is at least one of the reasons why some strains are able to produce spores in liquid-grown mycelium, while others cannot. By far the most conserved SALP is SsgB, which is most likely is the archetypal SALP, with only SsgB orthologues occurring in all morphologically complex actinomycetes. SsgB is essential for sporulation of Streptomyces, and orthologues have a universally conserved function in actinomycete morphogenesis. Show less
Microtubules are highly dynamic protein polymers that and are essential for intracellular organization and fundamental processes like transport and cell division. In cells, a wide family of... Show moreMicrotubules are highly dynamic protein polymers that and are essential for intracellular organization and fundamental processes like transport and cell division. In cells, a wide family of microtubule-associated proteins (MAPs) tightly regulates microtubule dynamics. The work presented in this thesis gives a high-resolution perspective on the microtubule assembly process and on the regulation mechanisms employed by representative MAPs. We studied dynamic microtubules outside cells, in a reconstituted minimal system. To follow microtubule growth with near molecular resolution, we developed a high-resolution technique that integrates optical tweezers, micro-fabricated rigid barriers and high-resolution video tracking of microbeads. Using this technique we found, for example, that microtubule assembly does not always occur by addition of single protein subunits, but multiple subunits could be incorporated at once at the growing end. XMAP215, a protein known to dramatically enhance microtubule growth, altered these molecular details. Another intriguing protein studied here is Mal3, a protein that is able to track growing microtubule ends. We found that Mal3 interacts differentially at the growing tip and on the rest of the microtubule, influencing all the parameters describing microtubule dynamics. In conclusion, our results give new insights into the microtubule assembly process in the absence and in the presence of regulators. Show less
Saccharomyces cerevisiae is one of the few yeast species that can grow equally well without molecular oxygen (anaerobic) as with this compound present (aerobic). This property has made it one of... Show moreSaccharomyces cerevisiae is one of the few yeast species that can grow equally well without molecular oxygen (anaerobic) as with this compound present (aerobic). This property has made it one of the most abundantly used yeasts in industry, since anaerobic incubation plays a major part in alcohol and bread industry. With the experiments described in this thesis it has been shown that apart from metabolic changes, an adaptation of the cell wall and the plasma membrane is very important for anaerobic growth. There seems to be a connection between this adaptation and the import of sterols, which are essential when no molecular oxygen is present. The importance of these adaptations becomes clear when the genomes of the facultative anaerobic Saccharomyces cerevisiae and the obligatory aerobic Kluyveromyces lactis are compared. K. lactis does not have any genes that encode for a sterol import system.The adaptation of the cell wall and the plasma membrane to anaerobic conditions is extensive and regulated in a complex way, as is apparent from the transcriptome data. Our experiments show that Snf7 is, at least in part, responsible for these changes. Show less
This work covers the evolution of satellite based Aeronautical Public Correspondence and the operational environment in which services are being offered. Followed by an examination of applicable... Show moreThis work covers the evolution of satellite based Aeronautical Public Correspondence and the operational environment in which services are being offered. Followed by an examination of applicable rules, including the relevant institutions from which they emanate, attention is devoted to questions on liability and the effect of State Sovereignty, applicable in territorial airspace, the Arctic, Antarctica, and the High Seas as well as, to activities such as telecommunications, air transport, copyright protection and trade in services. Particular attention is given to the ITU Radio Regulations; ITU Sector Recommendations; ICAO Council Regulations; International Copyright Laws; National Operational Procedures and Statutes. Show less
