Zeb2 DNA-Binding Sites in Neuroprogenitor Cells Reveal Autoregulation and Affirm Neurodevelopmental Defects, Including in Mowat-Wilson Syndrome

Functional perturbation and action mechanism studies have shown that the transcription factor Zeb2 controls cell fate decisions, differentiation, and/or maturation in multiple cell lineages in embryos and after birth. In cultured embryonic stem cells (ESCs), Zeb2’s mRNA/protein upregulation is necessary for the exit from primed pluripotency and for entering general and neural differentiation. We edited mouse ESCs to produce Flag-V5 epitope-tagged Zeb2 protein from one endogenous allele. Using chromatin immunoprecipitation coupled with sequencing (ChIP-seq), we mapped 2432 DNA-binding sites for this tagged Zeb2 in ESC-derived neuroprogenitor cells (NPCs). A new, major binding site maps promoter-proximal to Zeb2 itself. The homozygous deletion of this site demonstrates that autoregulation of Zeb2 is necessary to elicit the appropriate Zeb2-dependent effects in ESC-to-NPC differentiation. We have also cross-referenced all the mapped Zeb2 binding sites with previously obtained transcriptome data from Zeb2 perturbations in ESC-derived NPCs, GABAergic interneurons from the ventral forebrain of mouse embryos, and stem/progenitor cells from the post-natal ventricular-subventricular zone (V-SVZ) in mouse forebrain, respectively. Despite the different characteristics of each of these neurogenic systems, we found interesting target gene overlaps. In addition, our study also contributes to explaining developmental disorders, including Mowat-Wilson syndrome caused by ZEB2 deficiency, and also other monogenic syndromes.</p

Scientific maps should reach everyone: The cblindplot R package to let colour blind people visualise spatial patterns

Maps represent powerful tools to show the spatial variation of a variable in a straightforward manner. A crucial aspect in map rendering for its interpretation by users is the gamut of colours used for displaying data. One part of this problem is linked to the proportion of the human population that is colour blind and, therefore, highly sensitive to colour palette selection. The aim of this paper is to present the cblindplot R package and its founding function - cblind.plot() - which enables colour blind people to just enter an image in a coding workflow, simply set their colour blind deficiency type, and immediately get as output a colour blind friendly plot. We will first describe in detail colour blind problems, and then show a step by step example of the function being proposed. While examples exist to provide colour blind people with proper colour palettes, in such cases (i) the workflow include a separate import of the image and the application of a set of colour ramp palettes and (ii) albeit being well documented, there are many steps to be done before plotting an image with a colour blind friendly ramp palette. The function described in this paper, on the contrary, allows to (i) automatically call the image inside the function without any initial import step and (ii) explicitly refer to the colour blind deficiency type being experienced, to further automatically apply the proper colour ramp palette

Targeted chromatin conformation analysis identifies novel distal neural enhancers of ZEB2 in pluripotent stem cell differentiation

The transcription factor zinc finger E-box binding protein 2 (ZEB2) controls embryonic and adult cell fate decisions and cellular maturation in many stem/progenitor cell types. Defects in these processes in specific cell types underlie several aspects of Mowat-Wilson syndrome (MOWS), which is caused by ZEB2 haplo-insufficiency. Human ZEB2, like mouse Zeb2, is located on chromosome 2 downstream of a ±3.5 Mb-long gene-desert, lacking any protein-coding gene. Using temporal targeted chromatin capture (T2C), we show major chromatin structural changes based on mapping in-cis proximities between the ZEB2 promoter and this gene desert during neural differentiation of human-induced pluripotent stem cells, including at early neuroprogenitor cell (NPC)/rosette state, where ZEB2 mRNA levels increase significantly. Combining T2C with histone-3 acetylation mapping, we identified three novel candidate enhancers about 500 kb upstream of the ZEB2 transcription start site. Functional luciferase-based assays in heterologous cells and NPCs reveal co-operation between these three enhancers. This study is the first to document in-cis Regulatory Elements located in ZEB2's gene desert. The results further show the usability of T2C for future studies of ZEB2 REs in differentiation and maturation of multiple cell types and the molecular characterization of newly identified MOWS patients that lack mutations in ZEB2 protein-coding exons

Airborne Alternaria and Cladosporium Fungal Spores in Europe: Forecasting Possibilities and Relationships with Meteorological Parameters

Airborne fungal spores are prevalent components of bioaerosols with a large impact on ecology, economy and health. Their major socioeconomic effects could be reduced by accurate and timely prediction of airborne spore concentrations. The main aim of this study was to create and evaluate models of Alternaria and Cladosporium spore concentrations based on data on a continental scale. Additional goals included assessment of the level of generalization of the models in space and description of the main meteorological factors influencing fungal spore concentrations. Aerobiological monitoring was carried out at 18 sites in six countries across Europe over 3 to 21 years depending on site. Quantile random forest modelling was used to predict spore concentrations values. Generalization of the Alternaria and Cladosporium models was tested using (i) one model for all the sites, (ii) models for groups of sites, and (iii) models for individual sites. The study revealed the possibility of reliable prediction of fungal spore levels using gridded meteorological data. The classification models also showed the capacity for providing larger scale predictions of fungal spore concentrations. Regression models were distinctly less accurate than classification models due to several factors, including measurement errors and distinct day-to-day changes of concentrations. Temperature and vapour pressure proved to be the most important variables in the regression and classification models of Alternaria and Cladosporium spore concentrations. Accurate and operational daily-scale predictive models of bioaerosol abundances contribute to the assessment and evaluation of relevant exposure and consequently more timely and efficient management of phytopathogenic and of human allergic diseases

Airborne Alternaria and Cladosporium Fungal Spores in Europe: Forecasting Possibilities and Relationships with Meteorological Parameters

Airborne fungal spores are prevalent components of bioaerosols with a large impact on ecology, economy and health. Their major socioeconomic effects could be reduced by accurate and timely prediction of airborne spore concentrations. The main aim of this study was to create and evaluate models of Alternaria and Cladosporium spore concentrations based on data on a continental scale. Additional goals included assessment of the level of generalization of the models in space and description of the main meteorological factors influencing fungal spore concentrations. Aerobiological monitoring was carried out at 18 sites in six countries across Europe over 3 to 21 years depending on site. Quantile random forest modelling was used to predict spore concentrations values. Generalization of the Alternaria and Cladosporium models was tested using (i) one model for all the sites, (ii) models for groups of sites, and (iii) models for individual sites. The study revealed the possibility of reliable prediction of fungal spore levels using gridded meteorological data. The classification models also showed the capacity for providing larger scale predictions of fungal spore concentrations. Regression models were distinctly less accurate than classification models due to several factors, including measurement errors and distinct day-to-day changes of concentrations. Temperature and vapour pressure proved to be the most important variables in the regression and classification models of Alternaria and Cladosporium spore concentrations. Accurate and operational daily-scale predictive models of bioaerosol abundances contribute to the assessment and evaluation of relevant exposure and consequently more timely and efficient management of phytopathogenic and of human allergic diseases

Germinal center B cells recognize antigen through a specialized immune synapse architecture

B cell activation is regulated by B cell antigen receptor (BCR) signaling and antigen internalization in immune synapses. Using large-scale imaging across B cell subsets, we show that in contrast to naive and memory B cells, which gathered antigen towards the synapse center before internalization, germinal center (GC) B cells extracted antigen by a distinct pathway using small peripheral clusters. Both naive and GC B cell synapses required proximal BCR signaling, but GC cells signaled less through the protein kinase C-β (PKC-β)–NF-κB pathway and produced stronger tugging forces on the BCR, thereby more stringently regulating antigen binding. Consequently, GC B cells extracted antigen with better affinity discrimination than naive B cells, suggesting that specialized biomechanical patterns in B cell synapses regulate T-cell dependent selection of high-affinity B cells in GCs

Clonal replacement sustains long-lived germinal centers primed by respiratory viruses

Germinal centers (GCs) form in secondary lymphoid organs in response to infection and immunization and are the source of affinity-matured B cells. The duration of GC reactions spans a wide range, and long-lasting GCs (LLGCs) are potentially a source of highly mutated B cells. We show that rather than consisting of continuously evolving B cell clones, LLGCs elicited by influenza virus or SARS-CoV-2 infection in mice are sustained by progressive replacement of founder clones by naive-derived invader B cells that do not detectably bind viral antigens. Rare founder clones that resist replacement for long periods are enriched in clones with heavily mutated immunoglobulins, including some with very high affinity for antigen, that can be recalled by boosting. Our findings reveal underappreciated aspects of the biology of LLGCs generated by respiratory virus infection and identify clonal replacement as a potential constraint on the development of highly mutated antibodies within these structures

Cytoskeletal control of B cell responses to antigens.

The actin cytoskeleton is essential for cell mechanics and has increasingly been implicated in the regulation of cell signalling. In B cells, the actin cytoskeleton is extensively coupled to B cell receptor (BCR) signalling pathways, and defects of the actin cytoskeleton can either promote or suppress B cell activation. Recent insights from studies using single-cell imaging and biophysical techniques suggest that actin orchestrates BCR signalling at the plasma membrane through effects on protein diffusion and that it regulates antigen discrimination through the biomechanics of immune synapses. These mechanical functions also have a role in the adaptation of B cell subsets to specialized tasks during antibody responses

Państwo, gospodarka, społeczeństwo w integrującej się Europie TOM 3

Ze wstępu: "1 maja 2004 przyniesie radykalną zmianą sytuacji dotychczasowych kandydatów do Unii Europejskiej. Z roli aplikanta i petenta przekształcą się we współdecydenta. Już dziś z przyszłymi członkami konsultuje się większość kwestii wymagających strategicznych decyzji. Przez ostatnie dziesięć lat wysiłek polityczny i intelektualny był skierowany na uzyskanie członkostwa Unii, a w ostatnim okresie negocjacji - na osiągnięcie najlepszych według polityków i ekonomistów warunków akcesji. 1 ten etap mamy już za sobą. Pora zacząć patrzeć przed siebie, lecz niejako petent, ale kraj współodpowiedzialny za dalsze funkcjonowanie i rozwój powiększonej Unii. Z tej perspektywy istotnajest analiza gospodarki europejskiej, z którąjuż dziś gospodarka państw kandydackich, także Polski, jest silnie powiązana. Wiedza na ten temat jest uboga i ograniczona do przeglądu bieżących wskaźników makroekonomicznych. Zarówno w ośrodkach rządowych, jak i pozarządowych dominuje podejście analizujące, co z konkretnego wydarzenia w innym kraju wynika dla gospodarki polskiej. Stanowczo nie wystarczy to do pełnienia odpowiedzialnej roli współdecydenta. Potrzebna jest pogłębiona wiedza na temat gospodarki europejskiej jako całości i poszczególnych krajów, a także najważniejszych partnerów handlowych i gospodarczych zjednoczonej Europy. Konieczne są pogłębione prace studialne dotyczące mechanizmów międzynarodowych, gdyż organy unijne będą się zajmować w najbliższych latach dalszym rozwojem europejskiego jednolitego Rynku, rywalizacją gospodarczą z USA i krajami azjatyckimi, liberalizacjąhandlu światowego."(...