3,232 research outputs found
An Experimental Investigation Into the Performance of a Flush Water-Jet Inlet
An experimental investigation of the flow within a generic flush type water-jet inlet has
been carried out to identify the principal flow features and provide a basis for development
of computational fluid dynamics (CFD) models. Tests were performed in a
cavitation tunnel with the model inlet fitted to the test section ceiling, and effects of
thickening the ingested tunnel wall boundary layer were investigated. The model was
fitted with a range of instrumentation to investigate the ramp pressure distribution and
boundary layer development, lip incidence, and pump face flow properties. Observations
of lip and duct cavitation inception and behavior were also made. The results
showed the inlet performance to be generally improved with the ingestion of a thicker
boundary layer. The thickened boundary layer significantly reduced ramp boundary
layer separation and distortion of flow at the notional pump face. However, a greater
range of lip incidence occurred with the thickened boundary layer with consequent
greater likelihood of lip separation and cavitation occurrence. Ideal lip incidence and
pump face flow uniformity occurred at flow parameters significantly different from
those for ideal pump face pressure recovery. Large developed cavities on the inlet lip
were observed for a range of conditions typical of conventional high-speed vessel
operation
Xist and Tsix transcription dynamics is regulated by the X-to-autosome ratio and semistable transcriptional states
In female mammals, X chromosome inactivation (XCI) is a key process in the control of gene dosage compensation between Xlinked genes and autosomes. Xist and Tsix, two overlapping antisense-transcribed noncoding genes, are central elements of the X inactivation center (Xic) regulating XCI. Xist upregulation results in the coating of the entire X chromosome by Xist RNA in cis, whereas Tsix transcription acts as a negative regulator of Xist. Here, we generated Xist and Tsix reporter mouse embryonic stem (ES) cell lines to study the genetic and dynamic regulation of these genes upon differentiation. Our results revealed mutually antagonistic roles for Tsix on Xist and vice versa and indicate the presence of semistable transcriptional states of the Xic locus predicting the outcome of XCI. These transcriptional states are instructed by the X-t
Wnt3a protein reduces growth factor-driven expansion of human hematopoietic stem and progenitor cells in serum-free cultures
Abstract
Ex vivo expansion of hematopoietic stem and progenitor cells (HSPC) is a promising approach
to improve insufficient engraftment after umbilical cord blood stem cell transplantation
(UCB-SCT). Although culturing HSPC with hematopoietic cytokines results in robust
proliferation, it is accompanied with extensive differentiation and loss of self-renewal capacity.
Wnt signaling has been implicated in regulating HSPC fate decisions in vivo and in promoting
HSPC self-renewal by inhibition of differentiation, but the effects of Wnt on the ex
vivo expansion of HSPC are controversial. Here, we demonstrate that exogenous Wnt3a
protein suppresses rather than promotes the expansion of UCB-derived CD34+ cells in
serum free expansion cultures. The reduced expansion was also observed in cultures initiated
with LinCD34+
CD38lowCD45RACD90+
cells which are highly enriched in HSC and
was also observed in response to activation of beta-catenin signaling by GSK3 inhibition.
The presence of Wnt3a protein during the culture reduced the frequency of multilineage
CFU-GEMM and the long-term repopulation ability of the expanded HSPC. These data suggest
that Wnt signaling reduces expansion of human HSPC in growth factor-driven expansion
cultures by promoting differentiation of HSPC
Sparsest factor analysis for clustering variables: a matrix decomposition approach
We propose a new procedure for sparse factor analysis (FA) such that each variable loads only one common factor. Thus, the loading matrix has a single nonzero element in each row and zeros elsewhere. Such a loading matrix is the sparsest possible for certain number of variables and common factors. For this reason, the proposed method is named sparsest FA (SSFA). It may also be called FA-based variable clustering, since the variables loading the same common factor can be classified into a cluster. In SSFA, all model parts of FA (common factors, their correlations, loadings, unique factors, and unique variances) are treated as fixed unknown parameter matrices and their least squares function is minimized through specific data matrix decomposition. A useful feature of the algorithm is that the matrix of common factor scores is re-parameterized using QR decomposition in order to efficiently estimate factor correlations. A simulation study shows that the proposed procedure can exactly identify the true sparsest models. Real data examples demonstrate the usefulness of the variable clustering performed by SSFA
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Lipid-mediated Wnt protein stabilization enables serum-free culture of human organ stem cells
Wnt signalling proteins are essential for culture of human organ stem cells in organoids, but most Wnt protein formulations are poorly active in serum-free media. Here we show that purified Wnt3a protein is ineffective because it rapidly loses activity in culture media due to its hydrophobic nature, and its solubilization requires a detergent, CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), that interferes with stem cell self-renewal. By stabilizing the Wnt3a protein using phospholipids and cholesterol as carriers, we address both problems: Wnt activity remains stable in serum-free media, while non-toxic carriers allow the use of high Wnt concentrations. Stabilized Wnt3a supports strongly increased self-renewal of organ and embryonic stem cells and the serum-free establishment of human organoids from healthy and diseased intestine and liver. Moreover, the lipophilicity of Wnt3a protein greatly facilitates its purification. Our findings remove a major obstacle impeding clinical applications of adult stem cells and offer advantages for all cell culture uses of Wnt3a protein
Insights from Amphioxus into the Evolution of Vertebrate Cartilage
Central to the story of vertebrate evolution is the origin of the vertebrate head, a problem difficult to approach using paleontology and comparative morphology due to a lack of unambiguous intermediate forms. Embryologically, much of the vertebrate head is derived from two ectodermal tissues, the neural crest and cranial placodes. Recent work in protochordates suggests the first chordates possessed migratory neural tube cells with some features of neural crest cells. However, it is unclear how and when these cells acquired the ability to form cellular cartilage, a cell type unique to vertebrates. It has been variously proposed that the neural crest acquired chondrogenic ability by recruiting proto-chondrogenic gene programs deployed in the neural tube, pharynx, and notochord. To test these hypotheses we examined the expression of 11 amphioxus orthologs of genes involved in neural crest chondrogenesis. Consistent with cellular cartilage as a vertebrate novelty, we find that no single amphioxus tissue co-expresses all or most of these genes. However, most are variously co-expressed in mesodermal derivatives. Our results suggest that neural crest-derived cartilage evolved by serial cooption of genes which functioned primitively in mesoderm
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