The actin-binding protein profilin 2 is a novel regulator of iron homeostasis

Cellular iron homeostasis is controlled by the iron regulatory proteins (IRPs) 1 and 2 that bind cis-regulatory iron-responsive elements (IRE) on target messenger RNAs (mRNA). We identified profilin 2 (Pfn2) mRNA, which encodes an actin-binding protein involved in endocytosis and neurotransmitter release, as a novel IRP-interacting transcript, and studied its role in iron metabolism. A combination of electrophoretic mobility shift assay experiments and bioinformatic analyses led to the identification of an atypical and conserved IRE in the 39 untranslated region of Pfn2 mRNA. Pfn2 mRNA levels were significantly reduced in duodenal samples from mice with intestinal IRP ablation, suggesting that IRPs exert a positive effect on Pfn2 mRNA expression in vivo. Overexpression of Pfn2 in HeLa and Hepa1-6 cells reduced their metabolically active iron pool. Importantly, Pfn2-deficient mice showed iron accumulation in discrete areas of the brain (olfactory bulb, hippocampus, and midbrain) and reduction of the hepatic iron store without anemia. Despite low liver iron levels, hepatic hepcidin expression remained high, likely because of compensatory activation of hepcidin by mild inflammation. Splenic ferroportin was increased probably to sustain hematopoiesis. Overall, our results indicate that Pfn2 expression is controlled by the IRPs in vivo and that Pfn2 contributes to maintaining iron homeostasis in cell lines and mice

Non-neoclassical up/down asymmetry of impurity emission on Alcator C-Mod

We demonstrate that existing theories are insufficient to explain up/down asymmetries of argon x-ray emission in Alcator C-Mod ohmic plasmas. Instead of the poloidal variation, ñ[subscript z]/〈n[subscript z]〉, being of order the inverse aspect ratio, ϵ, and scaling linearly with B[subscript t][superscript _ over n][subscript e]/I[2 over p], it is observed over 0.8 < r/a < 1.0 to be of order unity and exhibits a threshold behaviour between 3.5 <B[subscript t][superscript _ over n][subscript e]/I[subscript p] < 4.0 (T10[superscript 20] m[superscript −3] MA[superscript −1]). The transition from a poloidally symmetric to asymmetric impurity distribution is shown to occur at densities just below those that trigger a reversal of the core toroidal rotation direction, thought to be linked to the transition between the linear and saturated ohmic confinement regimes. A possible drive is discussed by which anomalous radial transport might sustain the impurity density asymmetry as the ratio of the perpendicular to parallel equilibration times, τ[subscript ⊥,z]/τ[subscript ∥,z], approaches unity. This explanation requires a strong up/down asymmetry in radial flux which, while not observable on C-Mod, has been measured in TEXT and Tore Supra ohmic plasmas.United States. Dept. of Energy (Contract DE-FC02-99ER54512)United States. Dept. of Energy (Fusion Research Postdoctoral Research Program

Perpendicular momentum injection by lower hybrid wave in a tokamak

The injection of lower hybrid waves for current drive into a tokamak affects the profile of intrinsic rotation. In this article, the momentum deposition by the lower hybrid wave on the electrons is studied. Due to the increase in the poloidal momentum of the wave as it propagates into the tokamak, the parallel momentum of the wave increases considerably. The change of the perpendicular momentum of the wave is such that the toroidal angular momentum of the wave is conserved. If the perpendicular momentum transfer via electron Landau damping is ignored, the transfer of the toroidal angular momentum to the plasma will be larger than the injected toroidal angular momentum. A proper quasilinear treatment proves that both perpendicular and parallel momentum are transferred to the electrons. The toroidal angular momentum of the electrons is then transferred to the ions via different mechanisms for the parallel and perpendicular momentum. The perpendicular momentum is transferred to ions through an outward radial electron pinch, while the parallel momentum is transferred through collisions.Comment: 22 pages, 4 figure

Investigation of the Thomson scattering-ECE discrepancy in ICRF heated plasmas at Alcator C-Mod

This paper reports on new experiments at Alcator C-Mod that were performed in order to investigate the long-standing, unresolved discrepancy between Thomson scattering (TS) and electron cyclotron emission (ECE) measurements of electron temperature in high temperature tokamak plasmas. Ion cyclotron range of frequency (ICRF) heating is used to produce high temperature conditions where the type of TS-ECE discrepancy observed in the past at JET and TFTR should become observable. At Alcator C-Mod, plasmas with Te(0) up to 8 keV are obtained using ion cyclotron resonance heating (ICRH), ICRF mode conversion heating and a combination of the two heating methods in order to explore the hypothesis that the presence of ICRH-generated fast ions may be related to the discrepancy. In all high temperature cases, the TS and ECE measurements of electron temperature agree to within experimental uncertainties. We find no evidence for the type of discrepancy reported at JET and TFTR. These results show that the TS-ECE discrepancy does not depend on high temperatures alone and also that the presence of ICRH-generated fast ions is insufficient to cause the TS-ECE discrepancy.United States. Dept. of Energy (DE-FC02-99ER54512

Meaningful Effect Sizes, Intraclass Correlations, and Proportions of Variance Explained by Covariates for Planning Two- and Three-Level Cluster Randomized Trials of Social and Behavioral Outcomes

BACKGROUND: There is a need for greater guidance regarding design parameters and empirical benchmarks for social and behavioral outcomes to inform assumptions in the design and interpretation of cluster randomized trials (CRTs). OBJECTIVES: We calculated the empirical reference values on critical research design parameters associated with statistical power for children's social and behavioral outcomes, including effect sizes, intraclass correlations (ICCs), and proportions of variance explained by a covariate at different levels (R 2). SUBJECTS: Children from kindergarten to Grade 5 in the samples from four large CRTs evaluating the effectiveness of two classroom- and two school-level preventive interventions. MEASURES: Teacher ratings of students' social and behavioral outcomes using the Teacher Observation of Classroom Adaptation-Checklist and the Social Competence Scale-Teacher. RESEARCH DESIGN: Two types of effect size benchmarks were calculated: (1) normative expectations for change and (2) policy-relevant demographic performance gaps. The ICCs and R 2 were calculated using two-level hierarchical linear modeling (HLM), where students are nested within schools, and three-level HLM, where students were nested within classrooms, and classrooms were nested within schools. RESULTS AND CONCLUSIONS: Comprehensive tables of benchmarks and ICC values are provided to inform prevention researchers in interpreting the effect size of interventions and conduct power analyses for designing CRTs of children's social and behavioral outcomes. The discussion also provides a demonstration for how to use the parameter reference values provided in this article to calculate the sample size for two- and three-level CRTs designs

Hydrazones and Thiosemicarbazones Targeting Protein-Protein-Interactions of SARS-CoV-2 Papain-like Protease

The papain-like protease (PLpro) of SARS-CoV-2 is essential for viral propagation and, additionally, dysregulation of the host innate immune system. Using a library of 40 potential metal-chelating compounds we performed an X-ray crystallographic screening against PLpro. As outcome we identified six compounds binding to the target protein. Here we describe the interaction of one hydrazone (H1) and five thiosemicarbazone (T1-T5) compounds with the two distinct natural substrate binding sites of PLpro for ubiquitin and ISG15. H1 binds to a polar groove at the S1 binding site by forming several hydrogen bonds with PLpro. T1-T5 bind into a deep pocket close to the polyubiquitin and ISG15 binding site S2. Their interactions are mainly mediated by multiple hydrogen bonds and further hydrophobic interactions. In particular compound H1 interferes with natural substrate binding by sterical hindrance and induces conformational changes in protein residues involved in substrate binding, while compounds T1-T5 could have a more indirect effect. Fluorescence based enzyme activity assay and complementary thermal stability analysis reveal only weak inhibition properties in the high micromolar range thereby indicating the need for compound optimization. Nevertheless, the unique binding properties involving strong hydrogen bonding and the various options for structural optimization make the compounds ideal lead structures. In combination with the inexpensive and undemanding synthesis, the reported hydrazone and thiosemicarbazones represent an attractive scaffold for further structure-based development of novel PLpro inhibitors by interrupting protein-protein interactions at the S1 and S2 site

Inference of Experimental Radial Impurity Transport on Alcator C-Mod: Bayesian Parameter Estimation and Model Selection

We present a fully Bayesian approach for the inference of radial profiles of impurity transport coefficients and compare its results to neoclassical, gyrofluid and gyrokinetic modeling. Using nested sampling, the Bayesian Impurity Transport InferencE (BITE) framework can handle complex parameter spaces with multiple possible solutions, offering great advantages in interpretative power and reliability with respect to previously demonstrated methods. BITE employs a forward model based on the pySTRAHL package, built on the success of the well-known STRAHL code [Dux, IPP Report, 2004], to simulate impurity transport in magnetically-confined plasmas. In this paper, we focus on calcium (Ca, Z=20) Laser Blow-Off injections into Alcator C-Mod plasmas. Multiple Ca atomic lines are diagnosed via high-resolution X-ray Imaging Crystal Spectroscopy and Vacuum Ultra-Violet measurements. We analyze a sawtoothing I-mode discharge for which neoclassical and turbulent (quasilinear and nonlinear) predictions are also obtained. We find good agreement in diffusion across the entire radial extent, while turbulent convection and density profile peaking are estimated to be larger in experiment than suggested by theory. Efforts and challenges associated with the inference of experimental pedestal impurity transport are discussed.Comment: 38 pages, 19 figures, submitted for publication in Nuclear Fusio

Automated macrophage counting in DLBCL tissue samples: a ROF filter based approach

BackgroundFor analysis of the tumor microenvironment in diffuse large B-cell lymphoma (DLBCL) tissue samples, it is desirable to obtain information about counts and distribution of different macrophage subtypes. Until now, macrophage counts are mostly inferred from gene expression analysis of whole tissue sections, providing only indirect information. Direct analysis of immunohistochemically (IHC) fluorescence stained tissue samples is confronted with several difficulties, e.g. high variability of shape and size of target macrophages and strongly inhomogeneous intensity of staining. Consequently, application of commercial software is largely restricted to very rough analysis modes, and most macrophage counts are still obtained by manual counting in microarrays or high power fields, thus failing to represent the heterogeneity of tumor microenvironment adequately.MethodsWe describe a Rudin-Osher-Fatemi (ROF) filter based segmentation approach for whole tissue samples, combining floating intensity thresholding and rule-based feature detection. Method is validated against manual counts and compared with two commercial software kits (Tissue Studio 64, Definiens AG, and Halo, Indica Labs) and a straightforward machine-learning approach in a set of 50 test images. Further, the novel method and both commercial packages are applied to a set of 44 whole tissue sections. Outputs are compared with gene expression data available for the same tissue samples. Finally, the ROF based method is applied to 44 expert-specified tumor subregions for testing selection and subsampling strategies.ResultsAmong all tested methods, the novel approach is best correlated with manual count (0.9297). Automated detection of evaluation subregions proved to be fully reliable. Comparison with gene expression data obtained for the same tissue samples reveals only moderate to low correlation levels. Subsampling within tumor subregions is possible with results almost identical to full sampling. Mean macrophage size in tumor subregions is 152.5111.3 m(2).ConclusionsROF based approach is successfully applied to detection of IHC stained macrophages in DLBCL tissue samples. The method competes well with existing commercial software kits. In difference to them, it is fully automated, externally repeatable, independent on training data and completely documented. Comparison with gene expression data indicates that image morphometry constitutes an independent source of information about antibody-polarized macrophage occurence and distribution

Divertor heat flux challenge and mitigation in SPARC

Owing to its high magnetic field, high power, and compact size, the SPARC experiment will operate with divertor conditions at or above those expected in reactor-class tokamaks. Power exhaust at this scale remains one of the key challenges for practical fusion energy. Based on empirical scalings, the peak unmitigated divertor parallel heat flux is projected to be greater than 10 GW m-2. This is nearly an order of magnitude higher than has been demonstrated to date. Furthermore, the divertor parallel Edge-Localized Mode (ELM) energy fluence projections (∼11-34 MJ m-2) are comparable with those for ITER. However, the relatively short pulse length (∼25 s pulse, with a ∼10 s flat top) provides the opportunity to consider mitigation schemes unsuited to long-pulse devices including ITER and reactors. The baseline scenario for SPARC employs a ∼1 Hz strike point sweep to spread the heat flux over a large divertor target surface area to keep tile surface temperatures within tolerable levels without the use of active divertor cooling systems. In addition, SPARC operation presents a unique opportunity to study divertor heat exhaust mitigation at reactor-level plasma densities and power fluxes. Not only will SPARC test the limits of current experimental scalings and serve for benchmarking theoretical models in reactor regimes, it is also being designed to enable the assessment of long-legged and X-point target advanced divertor magnetic configurations. Experimental results from SPARC will be crucial to reducing risk for a fusion pilot plant divertor design

Radiative heat exhaust in Alcator C-Mod I-mode plasmas

In order to more completely demonstrate the I-mode regime as a compelling fusion reactor operating scenario, the first dedicated attempts at I-mode radiative heat exhaust and detachment were carried out on Alcator C-Mod. Results conclusively show that within the parameter space explored, an I/L back-transition is triggered prior to meaningful reductions in parallel heat flux, q||, target temperature, Te;tar, and target pressure, pe;tar, at the outer divertor. The exact mechanism for the I/L trigger remains uncertain, but a multi-diagnostic investigation suggests the pedestal regulation physics is impacted promptly by small amounts of N2 seeded into the private flux region. The time delay between when N2 contacts the plasma and the I/L transition is triggered varied from 30-120 ms, approximately 0.7-3 x tE, and the delay varied inversely with I-mode pedestal-top pressure, pe;95. Power and nitrogen influx scans indicate that the I/L transitions are not linked to excessive bulk-plasma impurity radiation. It is also shown that in the subsequent L-mode following nitrogen seeding, q|| and Te;tar can be reduced by factors of ~10. The I/L transition and L-mode exhaust results using N2 are compared to similar attempts using Ne where such q|| and Te;tar reductions in L-mode are limited to factors of 2-3. Implications for the I-mode regime are discussed, including needs for follow-up experiments on other facilities