Differentiation has evolved into a systematic feature of European integration. Still, EU member states have been eager to maintain unity and not let differentiated integration (DI) be reflected in... Show moreDifferentiation has evolved into a systematic feature of European integration. Still, EU member states have been eager to maintain unity and not let differentiated integration (DI) be reflected in institutions and processes of EU governance. The sovereign debt crisis was a turning point, triggering an unprecedented reinforcement of euro area specific institutions and policies. So far, few contributions have studied the institutional implications of DI in the EU. This dissertation examines whether and, if so, how and with what implications DI has been institutionalised in EMU governance. It analyses the organisational inclusiveness of crisis management structures, the evolution of the Eurogroup and Eurogroup Working Group, procedural norms and elite ideas. The findings suggest that the distinction between euro area insiders and outsiders has become embedded in how member state representatives perceive political reality, determine mutual role expectations and organise governance processes. While this institutionalisation of DI in EMU was facilitated by particular circumstances of the sovereign debt crisis, its implications are evident in EMU governance and reform processes until today. The dissertation makes theoretical and empirical contributions to account for DI in institutional and political developments in EMU and, thus, aims to enrich institutionalist debates in EU studies. Show less
Emilie Prast en Marian Hickendorff van de Universiteit Leiden verdiepen in hun keynote het thema differentiatie vanuit een meer vakdidactische insteek. Vertrekkend vanuit algemene principes van... Show moreEmilie Prast en Marian Hickendorff van de Universiteit Leiden verdiepen in hun keynote het thema differentiatie vanuit een meer vakdidactische insteek. Vertrekkend vanuit algemene principes van differentiatie voor verschillende doelgroepen zoomen zij in op differentiatie voor sterke rekenaars. Je krijgt inzicht in het belang van differentiatie voor sterke rekenaars, manieren waarop in de praktijk al wordt gedifferentieerd voor deze groep, en kansen om differentiatie voor sterke rekenaars te verbeteren. Daarbij krijg je tips om je rekenonderwijs systematisch af te stemmen op de onderwijsbehoeften van deze groep leerlingen. Show less
Van Geel, M.; Prast, E.J.; Keuning, T.; Luyten, H. 2023
Differentiated instruction concerns adapting education to students’ needs. However, relatively little is known about how students experience the degree to which their teachers fulfill their... Show moreDifferentiated instruction concerns adapting education to students’ needs. However, relatively little is known about how students experience the degree to which their teachers fulfill their individual educational needs. In the current study, the Differentiated Instruction from Students’ Perspective (DISP) questionnaire was developed and administered among 458 students of grade 4 to 6 of 22 primary school teachers in the Netherlands. The internal consistency of the DISP instrument was considered good. Students in general were moderately positive about the level of differentiation they experienced and few differences between DISP-ratings of students with varying mathematics achievement levels were identified. It is concluded that the DISP questionnaire is a suitable instrument to study students’ perspectives and can be used by researchers as well as by teachers who would like to gain insights in and improve their differentiation practice. Show less
Differentiation and achievement grouping are frequently implemented practices to adapt education to students’ varying educational needs based on achievement level. Potential didactical and... Show moreDifferentiation and achievement grouping are frequently implemented practices to adapt education to students’ varying educational needs based on achievement level. Potential didactical and socioemotional advantages and disadvantages of these practices have been discussed in the literature. However, little is known about the perspective of students themselves. This study examined how students (N = 428) perceived differentiation and within-class homogeneous achievement grouping in primary mathematics education, with attention for potential differences between students of diverse achievement levels. Students of Grades 1, 3 and 5 completed a questionnaire about various differentiated mathematics activities and (if applicable) within-class achievement grouping. In line with the didactical perspective on differentiation, extended instruction and less difficult tasks were appreciated most by low-achieving students whereas more difficult tasks were appreciated most by high-achieving students. Students of all achievement groups had largely positive attitudes about achievement grouping and about their own achievement group. However, some differences between achievement groups were found, with less favourable results for students placed in low achievement groups. Students’ responses to open-ended questions provided additional insights into the reasons behind students’ evaluations of differentiation and achievement grouping. Differences between grade levels were also explored. Show less
Adapting education to students’ diverse educational needs is widely recognised as an important, but also complex aspect of effective teaching. In this chapter, we provide insight into how Dutch... Show moreAdapting education to students’ diverse educational needs is widely recognised as an important, but also complex aspect of effective teaching. In this chapter, we provide insight into how Dutch primary school teachers implement differentiation based on students’ current mathematics achievement level. We review evidence from four independent samples in which the same teacher self-assessment questionnaire was administered (N = 907 teachers in total), supplemented with qualitative data from various perspectives: external observers, students, and teachers. Based on these sources of information, we identify the following general patterns. Teachers generally implement achievement-based differentiation at least to some extent. That is, student achievement is monitored, and efforts are taken to adapt instruction or practice to students’ current achievement level. This is often organised using within-class homogeneous achievement groups. While low-achieving students regularly receive additional instruction, specific instruction for high-achieving students is uncommon. Refined, qualitative strategies to diagnose students’ individual educational needs and to adapt education to these individual needs are also used relatively infrequently. These relatively infrequently used strategies point to areas for improvement. Furthermore, the flexibility of within-class achievement groups seems to vary and deserves more attention in future research and practice. Show less
The advent of the technology to isolate or generate human pluripotent stem cells provided the potential to develop a wide range of human models that could enhance understanding of mechanisms... Show moreThe advent of the technology to isolate or generate human pluripotent stem cells provided the potential to develop a wide range of human models that could enhance understanding of mechanisms underlying human development and disease. These systems are now beginning to mature and provide the basis for the development of in vitro assays suitable to understand the biological processes involved in the multi-organ systems of the human body, and will improve strategies for diagnosis, prevention, therapies and precision medicine. Induced pluripotent stem cell lines are prone to phenotypic and genotypic changes and donor/clone dependent vari-ability, which means that it is important to identify the most appropriate characterization markers and quality control measures when sourcing new cell lines and assessing differentiated cell and tissue culture preparations for experimental work. This paper considers those core quality control measures for human pluripotent stem cell lines and evaluates the state of play in the development of key functional markers for their differentiated cell derivatives to promote assurance of reproducibility of scientific data derived from pluripotent stem cell-based systems. Show less
Grosch, M.; Ittermann, S.; Shaposhnikov, D.; Drukker, M.E. 2020
Membrane-free intracellular biocondensates are enclosures of proteins and nucleic acids that form by phase separation. Extensive ensembles of nuclear "membraneless organelles" indicate their... Show moreMembrane-free intracellular biocondensates are enclosures of proteins and nucleic acids that form by phase separation. Extensive ensembles of nuclear "membraneless organelles" indicate their involvement in genome regulation. Indeed, nuclear bodies have been linked to regulation of gene expression by formation of condensates made of chromatin and RNA processing factors. Important questions pertain to the involvement of membraneless organelles in determining cell identity through their cell-type-specific composition and function. Paraspeckles provide a prism to these questions because they exhibit striking cell-type-specific patterns and since they are crucial in embryogenesis. Here, we outline known interactions between paraspeckles and chromatin, and postulate how such interactions may be important in regulation of cell fate transitions. Moreover, we propose long non-coding RNAs (lncRNAs) as candidates for similar regulation because many form foci that resemble biocondensates and exhibit dynamic patterns during differentiation. Finally, we outline approaches that could ascertain how chromatin-associated membraneless organelles regulate cellular differentiation. Show less
Berg, S.P.H. van den; Pardieck, I.N.; Lanfermeijer, J.; Sauce, D.; Klenerman, P.; Baarle, D. van; Arens, R. 2019
Upon cytomegalovirus (CMV) infection, large T-cell responses are elicited that remain high or even increase over time, a phenomenon named memory T-cell inflation. Besides, the maintained robust T... Show moreUpon cytomegalovirus (CMV) infection, large T-cell responses are elicited that remain high or even increase over time, a phenomenon named memory T-cell inflation. Besides, the maintained robust T-cell response, CMV-specific T cells seem to have a distinctive phenotype, characterized by an advanced differentiation state. Here, we will review this special differentiation status by discussing the cellular phenotype based on the expression of CD45 isoforms, costimulatory, inhibitory and natural killer receptors, adhesion and lymphocyte homing molecules, transcription factors, cytokines and cytotoxic molecules. In addition, we focus on whether the differentiation state of CMV-specific CD8 T cells is unique in comparison with other chronic viruses and we will discuss the possible impact of factors such as antigen exposure and aging on the advanced differentiation status of CMV-specific CD8 T cells. Show less
The immune system of mammals is responsible for protecting our body against pathogensand foreign substances (antigens), and it consists of two discrete lines of defense. The firstline called innate... Show moreThe immune system of mammals is responsible for protecting our body against pathogensand foreign substances (antigens), and it consists of two discrete lines of defense. The firstline called innate immunity and provide a quick and nonspecific defense. The innate immunityincludes different cells types, such as mast cells, macrophages, neutrophils, eosinophils,dendritic cells and natural killer (NK) cells. The second line of defense called adaptive immunityresponds in an antigen-specific manner, and comprised of B and T lymphocyte cells.This thesis focuses on one of the signals which are known to play an important role duringHSC repopulation and T cell development that is Wnt signaling pathway. Dependingon the tissue/cell types (microenvironment) and specific class of Wnt proteins binding tothe corresponding receptors on the developing lymphocytes, two discrete downstreampathways will be activated namely canonical or non-canonical Wnt pathway. The main aimof this thesis is to dissect the roles of these two distinct pathways during hematopoiesisand lymphocyte development in murine as a physiologically relevant animal model. Show less
Diabetes mellitus is amongst the leading causes of morbidity and mortality worldwide. Insulin-producing pancreatic β-cells are central in establishing adequate glucose regulation and loss of... Show moreDiabetes mellitus is amongst the leading causes of morbidity and mortality worldwide. Insulin-producing pancreatic β-cells are central in establishing adequate glucose regulation and loss of functional β-cells results in the development of diabetes. Although it was previously thought that fully differentiated cells cannot change phenotype, murine studies recently indicated that mature β-cells can change identity into other islet cells under conditions of (metabolic) stress.We present a novel agarose based microwell culture system that can be used for aggregate formation of human or rodent islet cells. We show that this platform provides reproducible results to study aggregation of primary human islet cells. Using this culture system together with β-cell specific lineage tracing, we find that mature human β-cells can spontaneously lose their identity and convert into glucagon-containing α-cells. We then used human pancreatic tissue from donors with T2DM and matched controls to explore loss of β-cell identity in T2DM. We report that cells indicative of loss of β-cell identity are found more frequently in tissue samples from donors with a history of T2DM. Finally, we show that Pax4 and GLP-1 receptor agonists can partially prevent loss of identity β-cell in our ex vivo model. Show less
The research described in this thesis focuses on the use of both embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) to understand cardiac lineage development and disease. To... Show moreThe research described in this thesis focuses on the use of both embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) to understand cardiac lineage development and disease. To investigate the possibility of studying inherited cardiac diseases, we compared pluripotent stem cell-derived cardiomyocytes by investigating both a mouse and human model of a complex cardiac overlap syndrome caused by a mutation in the gene SCN5A. We demonstrated that both ESC- and iPSC-derived cardiomyocytes can recapitulate the characteristics of the disease. Furthermore this thesis describes a detailed protocol to differentiate human pluripotent stem cells to cardiomyocytes that was applied in the study comparing hESC- and hiPSC- derived cardiomyocytes at several time points during cardiac differentiation. We targeted fluorescent marker GFP to one allele of NKX2-5 in a human iPSC line that now matched a similar human ESC reporter line previously generated in the laboratory. This offered the opportunity to obtain cardiomyocytes and their precursors at different time points during the differentiation and determine the true degree of similarity between both pluripotent stem cell sources. Additionally the same hESC- and hiPSC-derived cardiomyocytes were compared to a unique set of foetal heart samples. Show less
It is critical to gain knowledge in the underlying mechanisms that control human cardiovascular developm ent, which helps us to understand the onset of congenital cardiovascular diseases, and to... Show moreIt is critical to gain knowledge in the underlying mechanisms that control human cardiovascular developm ent, which helps us to understand the onset of congenital cardiovascular diseases, and to develop optimal culture methods for efficient in vitro cardiomyocyte differentiation from hPSCs, which are of interest for final translational applications including screening and efficacy assays for disease modelling, drug discovery and development, personalized medicine, and perhaps the regeneration of cardiovascular tissues for therapeutic purposes. In this thesis, we show how genetic manipulation of human pluripotent stem cells (hPSCs), resulting in the genomic integration of a fluorescent protein encoding sequence at the locus of a key cardiac transcription factor, allows us to visualize and isolate early pre-cardiac progenitors subpopulations, and to study the molecular mechanisms involved in their further differentiation to cells of the cardiac lineage, including smooth muscle cells, endothelial cells, and cardiomyocyte subtypes. Show less
Prast, E.J.; Weijer-Bergsma, E. van de; Kroesbergen, E.H.; Luit, J.E.H. van 2016
Since the first reports of human induced pluripotent stem cells (hiPSC), the field of pluripotent stem cell (PSC) research has grown in leap and bounds, particularly in the area of (cardiac)... Show moreSince the first reports of human induced pluripotent stem cells (hiPSC), the field of pluripotent stem cell (PSC) research has grown in leap and bounds, particularly in the area of (cardiac) disease modeling. This is in part because it is fairly easy to produce cardiomyocytes from hPSC and also there are now more assays available which can determine phenotypic behavior. This thesis describes and discusses improvements to reprogramming technology as well as its use in cardiac development and disease modeling Show less
Many of the recognition molecules and mechanisms involved in immune responses have no bias towards external stimuli, but also sense and respond to pathological and physiological changes of non... Show moreMany of the recognition molecules and mechanisms involved in immune responses have no bias towards external stimuli, but also sense and respond to pathological and physiological changes of non infectious origin taking place within the body. Aiming at defining potential immuno-therapeutic strategies to treat human atherosclerosis, the focus of this work was the modulation of immune processes determinant of atherosclerosis lesion progression or cessation in mice, such as hematopoiesis, diapedesis and intravasation, leukocyte differentiation, cholesterol uptake apoptosis and cell survival. Modulation of these processes, by using bone marrow transplantation of hematopoietic stem cells with genetic deficiencies or over-expressing human or mouse engineered genes, demonstrated to alter the fate of atherosclerotic lesions at the balance between macrophage accumulation and lesion vulnerability versus resolution of inflammation and wound healing. This thesis demonstrates that processes responsible for the development and progression of atherosclerosis are dynamic and can be modulated to induce lesion stabilization and disease resolution. These results are promising for the development of novel therapeutics and challenge the current notion that atherosclerosis has a predetermined fate towards lesion vulnerability to rupture, which in humans results in thrombosis and clinical manifestations such myocardial infarction or stroke and sudden death. Show less
