DOT1L Methyltransferase Regulates Calcium Influx in Erythroid Progenitor Cells in Response to Erythropoietin

Erythropoietin (EPO) signaling plays a vital role in erythropoiesis by regulating proliferation and lineage-specific differentiation of murine hematopoietic progenitor cells (HPCs). An important downstream response of EPO signaling is calcium (Ca²⁺) influx, which is regulated by transient receptor potential channel (TRPC) proteins, particularly TRPC2 and TRPC6. While EPO induces Ca²⁺ influx through TRPC2, TRPC6 inhibits the function of TRPC2. Thus, interactions between TRPC2 and TRPC6 regulate the rate of Ca²⁺ influx in EPO-induced erythropoiesis. In this study, we observed that the expression of TRPC6 in KIT-positive erythroid progenitor cells was regulated by DOT1L. DOT1L is a methyltransferase that plays an important role in many biological processes during embryonic development including early erythropoiesis. We previously reported that Dot1l knockout (Dot1l^KO) HPCs in the yolk sac failed to develop properly, which resulted in lethal anemia. In this study, we detected a marked downregulation of Trpc6 gene expression in Dot1l^KO progenitor cells in the yolk sac compared to the wild type (WT). The promoter and the proximal regions of the Trpc6 gene locus exhibited an enrichment of H3K79 methylation, which is mediated solely by DOT1L. However, the expression of Trpc2, the positive regulator of Ca²⁺ influx, remained unchanged, resulting in an increased TRPC2/TRPC6 ratio. As the loss of DOT1L decreased TRPC6, which inhibited Ca²⁺ influx by TRPC2, Dot1l^KO HPCs in the yolk sac exhibited accelerated and sustained elevated levels of Ca²⁺ influx. Such heightened Ca²⁺ levels might have detrimental effects on the growth and proliferation of HPCs in response to EPO.     

Designing to Facilitate Browsing: A Look Back at the Hyperties Workstation Browser

Since browsing hypertext can present a formidable cognitive challenge, user interface design plays a major role in determining acceptability. In the Unix workstation version of Hyperties, a research-oriented prototype, we focussed on design features that facilitate browsing. We first give a general overview of Hyperties and its markup language. Customizable documents can be generated by the conditional text feature that enables dynamic and selective display of text and graphics. In addition we present: — an innovative solution to link identification: pop-out graphical buttons of arbitrary shape.

— application of pie menus to permit low cognitive load actions that reduce the distraction of common actions, such as page turning or window selection.

— multiple window selection strategies that reduce clutter and housekeeping effort. We preferred piles-of-tiles, in which standard-sized windows were arranged in a consistent pattern on the display and actions could be done rapidly, allowing users to concentrate on the contents.

     

Algorithmic issues for three-invariant hyperplastic Critical State models

Implicit stress integration and the consistent tangents are presented for Critical State hyperplasticity models which include a dependence on the third invariant of stress. An elliptical deviatoric yielding criterion [43] is incorporated within the family of geotechnical models first proposed by Collins and Hilder [8]. An alternative expression for the yield function is proposed and the consequences of different forms of that function are revealed in terms of the stability and efficiency of the stress return algorithm. Errors associated with the integration scheme are presented. It is shown how calibration of the two new material constants is achieved through examining one-dimesional consolidation tests and undrained triaxial compression data. Material point simulations of drained triaxial compression tests are then compared with established experimental results. Strain probe analyses are used to demonstrate the concepts of energy dissipation and stored plastic work along with the robustness of the integration method. Over twenty finite element boundary value problems are then simulated. These include single three-dimensional element tests, plane strain footing analyses and cavity expansion tests. The rapid convergence of the global Newton–Raphson procedure using the consistent tangent is demonstrated in small strain and finite deformation simulations.     

Quality assurance of PASADENA hyperpolarization for 13C biomolecules

Object  Define MR quality assurance procedures for maximal PASADENA hyperpolarization of a biological ¹³C molecular imaging reagent. Materials and methods  An automated PASADENA polarizer and a parahydrogen generator were installed. ¹³C enriched hydroxyethyl acrylate, 1-¹³C, 2,3,3-d₃ (HEA), was converted to hyperpolarized hydroxyethyl propionate, 1-¹³C, 2,3,3-d₃ (HEP) and fumaric acid, 1-¹³C, 2,3-d₂ (FUM) to hyperpolarized succinic acid, 1-¹³C, 2,3-d₂ (SUC), by reaction with parahydrogen and norbornadiene rhodium catalyst. Incremental optimization of successive steps in PASADENA was implemented. MR spectra and in vivo images of hyperpolarized ¹³C imaging agents were acquired at 1.5 and 4.7 T. Results  Application of quality assurance (QA) criteria resulted in incremental optimization of the individual steps in PASADENA implementation. Optimal hyperpolarization of HEP of P = 20% was achieved by calibration of the NMR unit of the polarizer (B ₀ field strength ± 0.002 mT). Mean hyperpolarization of SUC, P = [15.3 ± 1.9]% (N = 16) in D ₂O, and P = [12.8 ± 3.1]% (N = 12) in H ₂O, was achieved every 5–8 min (range 13–20%). An in vivo ¹³C succinate image of a rat was produced. Conclusion  PASADENA spin hyperpolarization of SUC to 15.3% in average was demonstrated (37,400 fold signal enhancement at 4.7 T). The biological fate of ¹³C succinate, a normally occurring cellular intermediate, might be monitored with enhanced sensitivity.     

Thermal management of power electronics modules packaged by a stacked-plate technique

The research presented in this paper is part of a multidisciplinary research program of the Center for Power Electronics Systems at Virginia Tech. The program supported by the Office of Naval Research focuses on the development of innovative technologies for packaging power electronics building blocks. The primary objective of this research is to improve package performance and reliability through thermal management, i.e., reducing device temperatures for a given power level. The task of thermal management involves considering trade-offs in the electrical design, package layout and geometry, materials selection and processing, manufacturing feasibility, and production cost. Based on the electrical design of a simple building block, samples of packaged modules, rated at 600 V and 3.3 kW, were fabricated using a stacked-plate technique, termed metal posts interconnected parallel plate structure (MPIPPS). The MPIPPS technique allows the power devices to be interconnected between two direct-bond copper substrates via the use of metal posts. Thermal modeling results on the MPIPPS packaged modules indicate that the new packaging technique offers a superior thermal management means for packaging power electronics modules.