The Leukemia-Associated Mllt10/Af10-Dot1l Are Tcf4/β-Catenin Coactivators Essential for Intestinal Homeostasis

The leukemia-associated Mllt10/Af10 and its partner the histone methyltransferase Dot1l are identified as Tcf4/β-catenin co-activators and shown to be essential for Wnt-driven endogenous gene expression, intestinal development and homeostasis

ENDOGLIN is dispensable for vasculogenesis, but required for vascular endothelial growth factor-induced angiogenesis

ENDOGLIN (ENG) is a co-receptor for transforming growth factor-β (TGF-β) family members that is highly expressed in endothelial cells and has a critical function in the development of the vascular system. Mutations in Eng are associated with the vascular disease known as hereditary hemorrhagic telangiectasia type l. Using mouse embryonic stem cells we observed that angiogenic factors, including vascular endothelial growth factor (VEGF), induce vasculogenesis in embryoid bodies even when Eng deficient cells or cells depleted of Eng using shRNA are used. However, ENG is required for the stem cell-derived endothelial cells to organize effectively into tubular structures. Consistent with this finding, fetal metatarsals isolated from E17.5 Eng heterozygous mouse embryos showed reduced VEGF-induced vascular network formation. Moreover, shRNA-mediated depletion and pharmacological inhibition of ENG in human umbilical vein cells mitigated VEGF-induced angiogenesis. In summary, we demonstrate that ENG is required for efficient VEGF-induced angiogenesis

Quantitative Impact Assessment of Sewer Condition on Health Risk

Due to a variety of contaminants in floodwater, exposure to urban pluvial flooding may pose a health risk to humans. In-sewer defects may cause increased pluvial flooding, possibly increasing health risks. This paper addresses the impact of in-sewer defects on urban pluvial flooding and, subsequently, on infection probabilities for humans. As such, it provides a necessary input for risk-informed sewer maintenance strategies in order to preserve the hydraulic performance of a sewer system. Critical locations in sewer networks can be safeguarded through detecting changes in hydraulic properties of the sewer system, by using monitoring equipment or alternative inspection methods. Two combined sewer systems in The Netherlands with different characteristics are studied. The catchment-wide average infection probability was calculated using Quantitative Microbial Risk Assessment (QMRA) and flooding frequencies from Monte Carlo simulations with a hydrodynamic model. For the studied catchments, it is concluded that the occurrence of flooding is significantly affected by sediment deposits and, consequently, the infection probability as well. The impact of sediment deposits on infection probabilities depends on sewer systems characteristics and varies within the catchment. The results in this paper also demonstrate that further research on the relationship between flood duration and infection probabilities is required

A method to identify the weakest link in urban drainage systems

Urban drainage systems are composed of subsystems. The ratio of the storage and discharge capacities of the subsystems determines the performance. The performance of the urban water system may deteriorate as a result of the change in the ratio of storage to discharge capacity due to aging, urbanisation and climate change. We developed the graph-based weakest link method (GBWLM) to analyse urban drainage systems. Flow path analysis from graph theory is applied instead of hydrodynamic model simulations to reduce the computational effort. This makes it practically feasible to analyse urban drainage systems with multi-decade rainfall series. We used the GBWLM to analyse the effect of urban water system aging and/or climate scenarios on flood extent and frequency. The case study shows that the results of the hydrodynamic models and the GBWLM are similar. The rainfall intensities of storm events are expected to increase by approximately 20% in the Netherlands due to climate change. For the case study, such an increase in load has little impact on the flood frequency and extent caused by gully pots and surface water. However, it could lead to a 50% increase in the storm sewer flood frequency and an increase in the extent of flooding. HIGHLIGHTS An analysis of urban water systems with multiyear rainfall series.; A combined analysis of subsystems of urban water subsystems.; A sensitivity analysis of urban water systems in consequence of aging or climate change.; Comparing flood frequency and flood extent caused by capacity reduction of urban water subsystems.; Flow path analysis from graph theory instead of hydrodynamic model simulations.

Severe nasal clefting and abnormal embryonic apoptosis in Alx3/Alx4 double mutant mice

A group of mouse aristaless-related genes has been implicated in functions in the development of the craniofacial skeleton. We have generated an Alx3 mutant allele in which the lacZ coding sequence is inserted in-frame in the Alx3 gene and the sequences encoding the conserved protein domains are deleted. Mice homozygous for this null allele are indistinguishable from wild-type mice. Compound mutants of Alx3 and Alx4, however, show severe craniofacial abnormalities that are absent in Alx4 single mutants. Alx3/Alx4 double mutant newborn mice have cleft nasal regions. Most facial bones and many other neural crest derived skull elements are malformed, truncated or even absent. The craniofacial defects in Alx3/Alx4 double mutant embryos become anatomically manifest around embryonic day 10.5, when the nasal processes appear to be abnormally positioned. This most probably leads to a failure of the medial nasal processes to fuse in the facial midline and subsequently to the split face phenotype. We detected a significant increase in apoptosis localised in the outgrowing frontonasal process in embryonic day 10.0 double mutant embryos, which we propose to be the underlying cause of the subsequent malformations

Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele

Cycling Lgr5+ stem cells fuel the rapid turnover of the adult intestinal epithelium. The existence of quiescent Lgr5+ cells has been reported, while an alternative quiescent stem cell population is believed to reside at crypt position +4. Here, we generated a novel Ki67RFP knock-in allele that identifies dividing cells. Using Lgr5-GFP;Ki67RFP mice, we isolated crypt stem and progenitor cells with distinct Wnt signaling levels and cell cycle features and generated their molecular signature using microarrays. Stem cell potential of these populations was further characterized using the intestinal organoid culture. We found that Lgr5high stem cells are continuously in cell cycle, while a fraction of Lgr5low progenitors that reside predominantly at +4 position exit the cell cycle. Unlike fast dividing CBCs, Lgr5low Ki67- cells have lost their ability to initiate organoid cultures, are enriched in secretory differentiation factors, and resemble the Dll1 secretory precursors and the label-retaining cells of Winton and colleagues. Our findings support the cycling stem cell hypothesis and highlight the cell cycle heterogeneity of early progenitors during lineage commitment

Mouse and human urothelial cancer organoids: A tool for bladder cancer research

Bladder cancer is a common malignancy that has a relatively poor outcome. Lack of culture models for the bladder epithelium (urothelium) hampers the development of new therapeutics. Here we present a long-term culture system of the normal mouse urothelium and an efficient culture system of human bladder cancer cells. These so-called bladder (cancer) organoids consist of 3D structures of epithelial cells that recapitulate many aspects of the urothelium. Mouse bladder organoids can be cultured efficiently and genetically manipulated with ease, which was exemplified by creating genetic knockouts in the tumor suppressors Trp53 and Stag2. Human bladder cancer organoids can be derived efficiently from both resected tumors and biopsies and cultured and passaged for prolonged periods. We used this feature of human bladder organoids to create a living biobank consisting of bladder cancer organoids derived from 53 patients. Resulting organoids were characterized histologically and functionally. Organoid lines contained both basal and luminal bladder cancer subtypes based on immunohistochemistry and gene expression analysis. Common bladder cancer mutations like TP53 and FGFR3 were found in organoids in the biobank. Finally, we performed limited drug testing on organoids in the bladder cancer biobank