Dynamic flow distortion investigation in an S-duct using DDES and SPIV data

The dynamic flow distortion generated within convoluted aero-engine intakes can affect the performance and operability of the engine. There is a need for a better understanding of the main flow mechanisms which promote flow distortion at the exit of S-shaped intakes. This paper presents a detailed analysis of the main coherent structures in an S-duct flow field based on a Delayed Detached Eddy Simulation (DDES). The DDES capability to capture the characteristics of the highly unsteady flow field is demonstrated against high resolution, synchronous Stereoscopic Particle Image Velocimetry (SPIV) measurements at the Aerodynamic Interface Plane (AIP). The flow field mechanisms responsible for the main AIP perturbations are identified. Clockwise and counter-clockwise stream-wise vortices are alternately generated around the separation region at a frequency of St=0.53, which promotes the swirl switching at the AIP. Spanwise vortices are also shed from the separation region at a frequency of St=1.06, and convect downstream along the separated centreline shear layer. This results in a vertical modulation of the main loss region and a fluctuation of the velocity gradient between the high and low velocity flow at the AIP

Advancing haematopoietic stem and progenitor cell biology through single-cell profiling.

Haematopoietic stem and progenitor cells (HSPCs) sit at the top of the haematopoietic hierarchy, and their fate choices need to be carefully controlled to ensure balanced production of all mature blood cell types. As cell fate decisions are made at the level of the individual cells, recent technological advances in measuring gene and protein expression in increasingly large numbers of single cells have been rapidly adopted to study both normal and pathological HSPC function. In this review we emphasise the importance of combining the correct computational models with single-cell experimental techniques, and illustrate how such integrated approaches have been used to resolve heterogeneities in populations, reconstruct lineage differentiation, identify regulatory relationships and link molecular profiling to cellular function.Research in the authors' laboratory is supported by Cancer Research UK, the Biotechnology and Biological Sciences Research Council, Bloodwise, the Leukemia and Lymphoma Society, a Wellcome Trust Strategic Award (Tracing Early Mammalian Lineage Decisions by Single Cell Genomics) and core support grants by the Wellcome Trust to the Cambridge Institute for Medical Research and Wellcome Trust – MRC Cambridge Stem Cell Institute. FKH and SN gratefully acknowledge the MRC for funding of their studentships.This is the author accepted manuscript. The final version is available from Wiley via http://dx.doi.org/10.1002/1873-3468.1223

Single-Cell Sequencing in Normal and Malignant Hematopoiesis.

Funder: Wellcome TrustHematopoiesis is one of the best studied adult stem-cell systems, with a differentiation hierarchy progressing from immature hematopoietic stem cells to over 10 distinct mature cell types. Recent technological breakthroughs now make it possible to define transcriptional profiles in thousands of individual cells. Facilitated by the wealth of prior data on cell purification and analysis strategies, hematopoiesis has been one of the earliest experimental systems to which many of these new single-cell sequencing technologies have been applied. In this review, the authors focus on recent studies, which have shed light on heterogeneity within individual populations as well as the relationships between populations, and also attempt to characterize the differences between normal and disease/perturbed states

“My journey map”: Developing a qualitative approach to mapping young people’s progress in residential rehabilitation

Young people with substance misuse issues are at risk of harm from significant negative health and life events. Contemporary research notes both a historical failure to recognize the unique needs of adolescents, and the ongoing need for dedicated adolescent treatment programs and outcome measures. It is concerning that there is so little literature assessing the quality, availability, and effectiveness of adolescent-focused treatment programs, and no adolescent-specific measurement tools centered on a young person’s progress in residential treatment. This article reports on the process of developing a qualitative approach to mapping progress in treatment over time. The research seeks to develop an approach that captures, at three points in time and from multiple viewpoints, the progress of young people in four residential rehabilitation services located in New South Wales and Western Australia, across several dimensions of the personal and social aspects of life. Our aim is to develop an approach that is accessible to the alcohol and other drug workforce, and that informs the development of a psychometrically robust quantitative measure of progress that is meaningful and useful both to practitioners and to the young people themselves

Index sorting resolves heterogeneous murine hematopoietic stem cell populations.

Recent advances in the cellular and molecular biology of single stem cells have uncovered significant heterogeneity in the functional properties of stem cell populations. This has prompted the development of approaches to study single cells in isolation, often performed using multiparameter flow cytometry. However, many stem cell populations are too rare to test all possible cell surface marker combinations, and virtually nothing is known about functional differences associated with varying intensities of such markers. Here we describe the use of index sorting for further resolution of the flow cytometric isolation of single murine hematopoietic stem cells (HSCs). Specifically, we associate single-cell functional assay outcomes with distinct cell surface marker expression intensities. High levels of both CD150 and EPCR associate with delayed kinetics of cell division and low levels of differentiation. Moreover, cells that do not form single HSC-derived clones appear in the 7AAD(dim) fraction, suggesting that even low levels of 7AAD staining are indicative of less healthy cell populations. These data indicate that when used in combination with single-cell functional assays, index sorting is a powerful tool for refining cell isolation strategies. This approach can be broadly applied to other single-cell systems, both to improve isolation and to acquire additional cell surface marker information.This work was supported by grants from Leukaemia and Lymphoma Research, the Medical Research Council, the National Institute for Health Research Cambridge Biomedical Research Centre, and core support grants by the Wellcome Trust to the Cambridge Institute for Medical Research and Wellcome Trust–MRC Cambridge Stem Cell Institute. DGK is the recipient of a Canadian Institutes of Health Research Postdoctoral Fellowship and a European Hematology Association non-clinical advanced research fellowship. The authors declare that they have no conflict of interest.This is the author accepted manuscript. The final version will be available from Elsevier at http://dx.doi.org/10.1016/j.exphem.2015.05.006

Delayed detached-eddy simulation and particle image velocimetry investigation of S-Duct flow distortion

The dynamic flow distortion generated within convoluted aeroengine intakes can affect the performance and operability of the engine. There is a need for a better understanding of the main flow mechanisms that promote flow distortion at the exit of S-shaped intakes. This paper presents a detailed analysis of the main coherent structures in an S-duct flowfield based on a delayed detached-eddy simulation. The capability of this numerical approach to capture the characteristics of the highly unsteady flowfield is demonstrated against high-resolution, synchronous stereoscopic particle image velocimetry measurements at the aerodynamic interface plane. The flowfield mechanisms responsible for the main perturbations at the duct outlet are identified. Clockwise and counterclockwise streamwise vortices are alternately generated around the separation region at a frequency of St=0.53 St=0.53 , which promote the swirl switching at the duct outlet. Spanwise vortices are also shed from the separation region at a frequency of St=1.06 St=1.06 and convect downstream along the separated centerline shear layer. This results in a vertical modulation of the main loss region and a fluctuation of the velocity gradient between the high- and low-velocity flow at the aerodynamic interface plane

Gata3 targets Runx1 in the embryonic haematopoietic stem cell niche.

Runx1 is an important haematopoietic transcription factor as stressed by its involvement in a number of haematological malignancies. Furthermore, it is a key regulator of the emergence of the first haematopoietic stem cells (HSCs) during development. The transcription factor Gata3 has also been linked to haematological disease and was shown to promote HSC production in the embryo by inducing the secretion of important niche factors. Both proteins are expressed in several different cell types within the aorta-gonads-mesonephros (AGM) region, in which the first HSCs are generated; however, a direct interaction between these two key transcription factors in the context of embryonic HSC production has not formally been demonstrated. In this current study, we have detected co-localisation of Runx1 and Gata3 in rare sub-aortic mesenchymal cells in the AGM. Furthermore, the expression of Runx1 is reduced in Gata3 -/- embryos, which also display a shift in HSC emergence. Using an AGM-derived cell line as a model for the stromal microenvironment in the AGM and performing ChIP-Seq and ChIP-on-chip experiments, we demonstrate that Runx1, together with other key niche factors, is a direct target gene of Gata3. In addition, we can pinpoint Gata3 binding to the Runx1 locus at specific enhancer elements which are active in the microenvironment. These results reveal a direct interaction between Gata3 and Runx1 in the niche that supports embryonic HSCs and highlight a dual role for Runx1 in driving the transdifferentiation of haemogenic endothelial cells into HSCs as well as in the stromal cells that support this process.This work was supported by an Intermediate Fellowship (K.O.) and a Junior Fellowship (S.R.F.) from the Kay Kendall Leukaemia Fund, a British Society for Haematology Early Stage Investigator Fellowship (K.O.) as well as funding from Bloodwise (N.K.W. and B.G.), MRC (N.K.W. and B.G.) and the Wellcome Trust (N.K.W. and B.G.). MdB is funded by a programme in the MRC Molecular Hematology Unit Core award (Grant number: MC_UU_12009/2). Core facilities are supported by Strategic Award WT100140, equipment grant 093026 and centre grant MR/K017047/1

Myeloid protein tyrosine phosphatase 1B (PTP1B) deficiency protects against atherosclerotic plaque formation in the ApoE-/- mouse model of atherosclerosis with alterations in IL10/AMPKa pathway

Objective: Cardiovascular disease (CVD) is the most prevalent cause of mortality among patients with Type 1 or Type 2 diabetes, due to accelerated atherosclerosis. Recent evidence suggests a strong link between atherosclerosis and insulin resistance due to impaired insulin receptor (IR) signaling. Moreover, inflammatory cells, in particular macrophages, play a key role in pathogenesis of atherosclerosis and insulin resistance in humans. We hypothesized that inhibiting the activity of protein tyrosine phosphatase 1B (PTP1B), the major negative regulator of the IR, specifically in macrophages, would have beneficial anti-inflammatory effects and lead to protection against atherosclerosis and CVD. Methods: We generated novel macrophage-specific PTP1B knockout mice on atherogenic background (ApoE−/−/LysM-PTP1B). Mice were fed standard or pro-atherogenic diet, and body weight, adiposity (echoMRI), glucose homeostasis, atherosclerotic plaque development, and molecular, biochemical and targeted lipidomic eicosanoid analyses were performed. Results: Myeloid-PTP1B knockout mice on atherogenic background (ApoE−/−/LysM-PTP1B) exhibited a striking improvement in glucose homeostasis, decreased circulating lipids and decreased atherosclerotic plaque lesions, in the absence of body weight/adiposity differences. This was associated with enhanced phosphorylation of aortic Akt, AMPKα and increased secretion of circulating anti-inflammatory cytokine interleukin-10 (IL-10) and prostaglandin E2 (PGE2), without measurable alterations in IR phosphorylation, suggesting a direct beneficial effect of myeloid-PTP1B targeting. Conclusions: Here we demonstrate that inhibiting the activity of PTP1B specifically in myeloid lineage cells protects against atherosclerotic plaque formation, under atherogenic conditions, in an ApoE−/− mouse model of atherosclerosis. Our findings suggest for the first time that macrophage PTP1B targeting could be a therapeutic target for atherosclerosis treatment and reduction of CVD risk

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Reflections and Experiences of a Co-Researcher involved in a Renal Research Study

Background Patient and Public Involvement (PPI) is seen as a prerequisite for health research. However, current Patient and public involvement literature has noted a paucity of recording of patient and public involvement within research studies. There have been calls for more recordings and reflections, specifically on impact. Renal medicine has also had similar criticisms and any reflections on patient and public involvement has usually been from the viewpoint of the researcher. Roles of patient and public involvement can vary greatly from sitting on an Advisory Group to analysing data. Different PPI roles have been described within studies; one being a co-researcher. However, the role of the co-researcher is largely undefined and appears to vary from study to study. Methods The aims of this paper are to share one first time co-researcher's reflections on the impact of PPI within a mixed methods (non-clinical trial) renal research study. A retrospective, reflective approach was taken using data available to the co-researcher as part of the day-to-day research activity. Electronic correspondence and documents such as meeting notes, minutes, interview thematic analysis and comments on documents were re-examined. The co-researcher led on writing this paper. Results This paper offers a broad definition of the role of the co-researcher. The co-researcher reflects on undertaking and leading on the thematic analysis of interview transcripts, something she had not previously done before. The co-researcher identified a number of key themes; the differences in time and responsibility between being a coresearcher and an Advisory Group member; how the role evolved and involvement activities could match the co-researchers strengths (and the need for flexibility); the need for training and support and lastly, the time commitment. It was also noted that it is preferable that a co-researcher needs to be involved from the very beginning of the grant application. Conclusions The reflections, voices and views of those undertaking PPI has been largely underrepresented in the literature. The role of co-researcher was seen to be rewarding but demanding, requiring a large time commitment. It is hoped that the learning from sharing this experience will encourage others to undertake this role, and encourage researchers to reflect on the needs of those involved.Peer reviewedFinal Published versio