Self-renewal of single mouse hematopoietic stem cells is reduced by JAK2V617F without compromising progenitor cell expansion

Recent descriptions of significant heterogeneity in normal stem cells and cancers have altered our understanding of tumorigenesis, emphasizing the need to understand how single stem cells are subverted to cause tumors. Human myeloproliferative neoplasms (MPNs) are thought to reflect transformation of a hematopoietic stem cell (HSC) and the majority harbor an acquired V617F mutation in the JAK2 tyrosine kinase, making them a paradigm for studying the early stages of tumor establishment and progression. The consequences of activating tyrosine kinase mutations for stem and progenitor cell behavior are unclear. In this article, we identify a distinct cellular mechanism operative in stem cells. By using conditional knock-in mice, we show that the HSC defect resulting from expression of heterozygous human JAK2V617F is both quantitative (reduced HSC numbers) and qualitative (lineage biases and reduced self-renewal per HSC). The defect is intrinsic to individual HSCs and their progeny are skewed toward proliferation and differentiation as evidenced by single cell and transplantation assays. Aged JAK2V617F show a more pronounced defect as assessed by transplantation, but mice that transform reacquire competitive self-renewal ability. Quantitative analysis of HSC-derived clones was used to model the fate choices of normal and JAK2-mutant HSCs and indicates that JAK2V617F reduces self-renewal of individual HSCs but leaves progenitor expansion intact. This conclusion is supported by paired daughter cell analyses, which indicate that JAK2-mutant HSCs more often give rise to two differentiated daughter cells. Together these data suggest that acquisition of JAK2V617F alone is insufficient for clonal expansion and disease progression and causes eventual HSC exhaustion. Moreover, our results show that clonal expansion of progenitor cells provides a window in which collaborating mutations can accumulate to drive disease progression. Characterizing the mechanism(s) of JAK2V617F subclinical clonal expansions and the transition to overt MPNs will illuminate the earliest stages of tumor establishment and subclone competition, fundamentally shifting the way we treat and manage cancers

Taming the rugged energy landscape: Techniques for the production, reordering, and stabilization of selected cluster inherent structures

We report our studies of the potential energy surface (PES) of selected binary Lennard-Jones clusters. The effect of adding selected impurity atoms to a homogeneous cluster is explored. Inherent structures and transition states are found by combination of conjugate-gradient and eigenvector-following methods while the topography of the PES is mapped with the help of a disconnectivity analysis. We show that we can controllably induce new structures as well as reorder and stabilize existing structures that are characteristic of higher-lying minima.Comment: 9 pages, 9 figures, accepted for publication in J. Chem. Phy

“It’s my dream to work with Olympic athletes”: Neophyte sport psychologists’ expectations and initial experiences regarding service delivery

We examined trainee practitioners' initial experiences of applied sport psychology practice. Semi-structured interviews (4) were conducted over 6 months with 7 full-time MSc students before, during, and after the applied sport psychology module, when they were working with clients. Participants also kept reflective diaries over an 8-week period whilst working with clients. Findings included: (a) motivations and expectations of an ASP practice career, (b) perceptions of service delivery, (c) emotional demands, and (d) pivotal experiences. Findings extend previous literature on the initial stages of practitioner development, providing micro-level detail on aspects of the intense development process during this pivotal perio

Preclinical single photon emission computed tomography of alpha particle-emitting radium-223

Objective: Dose optimization and pharmacokinetic evaluation of α-particle emitting radium-223 dichloride (223RaCl2) by planar γ-camera or single photon emission computed tomography (SPECT) imaging are hampered by the low photon abundance and injected activities. In this study, we demonstrate SPECT of 223Ra using phantoms and small animal in vivo models. Methods: Line phantoms and mice bearing 223Ra were imaged using a dedicated small animal SPECT by detecting the low-energy photon emissions from 223Ra. Localization of the therapeutic agent was verified by whole-body and whole-limb autoradiography and its radiobiological effect confirmed by immunofluorescence. Results: A state-of-the-art commercial small animal SPECT system equipped with a highly sensitive collimator enables collection of sufficient counts for three-dimensional reconstruction at reasonable administered activities and acquisition times. Line sources of 223Ra in both air and in a water scattering phantom gave a line spread function with a full-width-at-half-maximum of 1.45 mm. Early and late-phase imaging of the pharmacokinetics of the radiopharmaceutical were captured. Uptake at sites of active bone remodeling was correlated with DNA damage from the α particle emissions. Conclusions: This work demonstrates the capability to noninvasively define the distribution of 223RaCl2, a recently approved α-particle-emitting radionuclide. This approach allows quantitative assessment of 223Ra distribution and may assist radiation-dose optimization strategies to improve therapeutic response and ultimately to enable personalized treatment planning

Demonstration of Multi-Gbps Data Rates at Ka-Band Using Software-Defined Modem and Broadband High Power Amplifier for Space Communications

The paper presents the first ever research and experimental results regarding the combination of a software-defined multi-Gbps modem and a broadband high power space amplifier when tested with an extended form of the industry standard DVB-S2 and LDPC rate 9/10 FEC codec. The modem supports waveforms including QPSK, 8-PSK, 16-APSK, 32-APSK, 64-APSK, and 128-QAM. The broadband high power amplifier is a space qualified traveling-wave tube (TWT), which has a passband greater than 3 GHz at 33 GHz, output power of 200 W and efficiency greater than 60 percent. The modem and the TWTA together enabled an unprecedented data rate at 20 Gbps with low BER of 10(exp -9). The presented results include a plot of the received waveform constellation, BER vs. E(sub b)/N(sub 0) and implementation loss for each of the modulation types tested. The above results when included in an RF link budget analysis show that NASA s payload data rate can be increased by at least an order of magnitude (greater than 10X) over current state-of-practice, limited only by the spacecraft EIRP, ground receiver G/T, range, and available spectrum or bandwidth

Peak Satellite-to-Earth Data Rates Derived From Measurements of a 20 Gbps Bread-Board Modem

A prototype data link using a Ka-band space qualified, high efficiency 200 W TWT amplifier and a bread-board modem emulator were created to explore the feasibility of very high speed communications in satellite-to-earth applications. Experiments were conducted using a DVB-S2-like waveform with modifications to support up to 20 Gbps through the addition of 128-Quadrature Amplitude Modulation (QAM). Limited by the bandwidth of the amplifier, a constant peak symbol rate of 3.2 Giga-symbols/sec was selected and the modulation order was varied to explore what peak data rate might be supported by an RF link through this amplifier. Using 128-QAM, an implementation loss of 3 dB was observed at 20 Gbps, and the loss decreased as data rate or bandwidth were reduced. Building on this measured data, realistic link budget calculations were completed. Low-Earth orbit (LEO) missions based on this TWTA with reasonable hardware assumptions and antenna sizing are found to be bandwidth-limited, rather than power-limited, making the spectral efficiency of 9/10-rate encoded 128-QAM very attractive. Assuming a bandwidth allocation of 1 GHz, these computations indicate that low-Earth orbit vehicles could achieve data rates up to 5 Gbps-an order of magnitude beyond the current state-of-practice, yet still within the processing power of a current FPGA-based software-defined modem. The measured performance results and a description of the experimental setup are presented to support these conclusions

IL-21 production by CD4+ effector T cells and frequency of circulating follicular helper T cells are increased in type 1 diabetes patients.

AIMS/HYPOTHESIS: Type 1 diabetes results from the autoimmune destruction of insulin-secreting pancreatic beta cells by T cells. Despite the established role of T cells in the pathogenesis of the disease, to date, with the exception of the identification of islet-specific T effector (Teff) cells, studies have mostly failed to identify reproducible alterations in the frequency or function of T cell subsets in peripheral blood from patients with type 1 diabetes. METHODS: We assessed the production of the proinflammatory cytokines IL-21, IFN-γ and IL-17 in peripheral blood mononuclear cells from 69 patients with type 1 diabetes and 61 healthy donors. In an additional cohort of 30 patients with type 1 diabetes and 32 healthy donors, we assessed the frequency of circulating T follicular helper (Tfh) cells in whole blood. IL-21 and IL-17 production was also measured in peripheral blood mononuclear cells (PBMCs) from a subset of 46 of the 62 donors immunophenotyped for Tfh. RESULTS: We found a 21.9% (95% CI 5.8, 40.2; p = 3.9 × 10(-3)) higher frequency of IL-21(+) CD45RA(-) memory CD4(+) Teffs in patients with type 1 diabetes (geometric mean 5.92% [95% CI 5.44, 6.44]) compared with healthy donors (geometric mean 4.88% [95% CI 4.33, 5.50]). Consistent with this finding, we found a 14.9% increase in circulating Tfh cells in the patients (95% CI 2.9, 26.9; p = 0.016). CONCLUSIONS/INTERPRETATION: These results indicate that increased IL-21 production is likely to be an aetiological factor in the pathogenesis of type 1 diabetes that could be considered as a potential therapeutic target.This work was supported by the JDRF UK Centre for Diabetes - Genes, Autoimmunity and Prevention (D-GAP; 4-2007-1003) in collaboration with M. Peakman and T. Tree at King’s College London, the JDRF, the Wellcome Trust (WT; WT061858/091157 and 083650/Z/07/Z) and the National Institute for Health Research Cambridge Biomedical Research Centre (CBRC). The Cambridge Institute for Medical Research (CIMR) is in receipt of a Wellcome Trust Strategic Award (100140). RCF is funded by a JDRF post-doctoral fellowship (3-2011-374). CW is funded by the Wellcome Trust (088998). The funding organisations had no involvement with the design and conduct of the study; collection,management, analysis, and interpretation of the data; and preparation, review, or approval of the manuscript.This is the final published version. It first appeared at http://link.springer.com/article/10.1007%2Fs00125-015-3509-8