Nuclear membrane disassembly and rupture

The nuclear envelope consists of two membranes traversed by nuclear pore complexes. The outer membrane is continuous with the endoplasmic reticulum. At mitosis nuclear pore complexes are dismantled and membranes disperse. The mechanism of dispersal is controversial: one view is that membranes feed into the endoplasmic reticulum, another is that they vesiculate. Using Xenopus egg extracts, nuclei have been assembled and then induced to breakdown by addition of metaphase extract. Field emission scanning electron microscopy was used to study disassembly. Strikingly, endoplasmic reticulum-like membrane tubules form from the nuclear surface after the addition of metaphase extracts, but vesicles were also observed. Microtubule inhibitors slowed but did not prevent membrane removal, whereas Brefeldin A, which inhibits vesicle formation, stops membrane disassembly, suggesting that vesiculation is necessary. Structures that looked like coated buds were observed and buds were labelled for β-COP. We show that nuclear pore complexes are dismantled and the pore closed prior to membrane rupturing, suggesting that rupturing is an active process rather than a result of enlargement of nuclear pores

Corporate Inversions: New challenges, New Opportunities

With a wave of recent tax inversion and corporate reorganization discussions, corporate tax strategy has begun to move to the forefront of media, public and Congressional attention. These high-profile inversion strategies have gained momentum and achieved heightened attention, becoming a matter of public policy matter in 2014. While corporate international tax strategies have existed since the dawn of the U.S. federal income tax, inversions in their current form have been active only since the 1980s. Using three predominate inversion cases as a lens, this research intends to fill a gap in the existing literature relating to corporate inversions. By combining existing case law, tax legislation, and Treasury regulations, this paper develops a framework for supporting strategic global tax initiatives. The conclusions and recommendations reached are generalizable and appropriate for use in developing best practice solutions

The Immune Adaptor SLP-76 Binds to SUMO-RANGAP1 at Nuclear Pore Complex Filaments to Regulate Nuclear Import of Transcription Factors in T Cells

While immune cell adaptors regulate proximal T cell signaling, direct regulation of the nuclear pore complex (NPC) has not been reported. NPC has cytoplasmic filaments composed of RanGAP1 and RanBP2 with the potential to interact with cytoplasmic mediators. Here, we show that the immune cell adaptor SLP-76 binds directly to SUMO-RanGAP1 of cytoplasmic fibrils of the NPC, and that this interaction is needed for optimal NFATc1 and NF-κB p65 nuclear entry in T cells. Transmission electron microscopy showed anti-SLP-76 cytoplasmic labeling of the majority of NPCs in anti-CD3 activated T cells. Further, SUMO-RanGAP1 bound to the N-terminal lysine 56 of SLP-76 where the interaction was needed for optimal RanGAP1-NPC localization and GAP exchange activity. While the SLP-76-RanGAP1 (K56E) mutant had no effect on proximal signaling, it impaired NF-ATc1 and p65/RelA nuclear entry and in vivo responses to OVA peptide. Overall, we have identified SLP-76 as a direct regulator of nuclear pore function in T cells

The major inducible small heat shock protein HSP20-3 in the tardigrade Ramazzottius varieornatus forms filament-like structures and is an active chaperone.

The tardigrade Ramazzottius varieornatus has remarkable resilience to a range of environmental stresses. In this study, we have characterised two members of the small heat shock protein (sHSP) family in R. varieornatus, HSP20-3 and HSP20-6. These are the most highly upregulated sHSPs in response to a 24 h heat shock at 35 C of adult tardigrades with HSP20-3 being one of the most highly upregulated gene in the whole transcriptome. Both R. varieornatus sHSPs and the human sHSP, CRYAB (HSPB5), were produced recombinantly for comparative structure-function studies. HSP20-3 exhibited a superior chaperone activity than human CRYAB in a heat-induced protein aggregation assay. Both tardigrade sHSPs also formed larger oligomers than CRYAB as assessed by size exclusion chromatography and transmission electron microscopy of negatively stained samples. Whilst both HSP20-3 and HSP20-6 formed particles that were variable in size and larger than the particles formed by CRYAB, only HSP20-3 formed filament-like structures. The particles and filament-like structures formed by HSP20-3 appear inter-related as the filament-like structures often had particles located at their ends. Sequence analyses identified two unique features; an insertion in the middle region of the N-terminal domain (NTD) and preceding the critical-sequence identified in CRYAB, as well as a repeated QNTN-motif located in the C-terminal domain of HSP20-3. The NTD insertion is expected to affect protein-protein interactions and subunit oligomerisation. Removal of the repeated QNTN-motif abolished HSP20-3 chaperone activity and also affected the assembly of the filament-like structures. We discuss the potential contribution of HSP20-3 to protein condensate formation. [Abstract copyright: Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.

Analysis of CMB polarization on an incomplete sky

The full sky cosmic microwave background polarization field can be decomposed into 'electric' and 'magnetic' components. Working in harmonic space we construct window functions that allow clean separation of the electric and magnetic modes from observations over only a portion of the sky. Our construction is exact for azimuthally symmetric patches, but should continue to perform well for arbitrary patches. From the window functions we obtain variables that allow for robust estimation of the magnetic component without risk of contamination from the probably much larger electric signal. For isotropic, uncorrelated noise the variables have a very simple diagonal noise correlation, and further analysis using them should be no harder than analysing the temperature field. For an azimuthally-symmetric patch, such as that obtained from survey missions when the galactic region is removed, the exactly-separated variables are fast to compute allowing us to estimate the magnetic signal that could be detected by the Planck satellite in the absence of non-galactic foregrounds. We also discuss the sensitivity of future experiments to tensor modes in the presence of a magnetic signal generated by weak lensing, and give lossless methods for analysing the electric polarization field in the case that the magnetic component is negligible.Comment: 27 pages, 8 figures. New appendix on weak signal detection and revised plots using a better statistic. Other changes to match version accepted by PRD. Sample source code now available at http://cosmologist.info/pola

Generating natural language specifications from UML class diagrams

Early phases of software development are known to be problematic, difficult to manage and errors occurring during these phases are expensive to correct. Many systems have been developed to aid the transition from informal Natural Language requirements to semistructured or formal specifications. Furthermore, consistency checking is seen by many software engineers as the solution to reduce the number of errors occurring during the software development life cycle and allow early verification and validation of software systems. However, this is confined to the models developed during analysis and design and fails to include the early Natural Language requirements. This excludes proper user involvement and creates a gap between the original requirements and the updated and modified models and implementations of the system. To improve this process, we propose a system that generates Natural Language specifications from UML class diagrams. We first investigate the variation of the input language used in naming the components of a class diagram based on the study of a large number of examples from the literature and then develop rules for removing ambiguities in the subset of Natural Language used within UML. We use WordNet,a linguistic ontology, to disambiguate the lexical structures of the UML string names and generate semantically sound sentences. Our system is developed in Java and is tested on an independent though academic case study

Ultrahigh-Energy Neutrino-Nucleon Cross Sections and Perturbative Unitarity

Unitarity relates the total cross section for neutrino-nucleon scattering to the neutrino-nucleon forward scattering amplitude. Assuming the validity of the perturbative expansion of the forward amplitude in the {\em weak} coupling constant, we derive a unitarity bound on the inelastic cross section. The inelastic cross section saturates this bound at a typical neutrino energy $E_\nu \simeq 10^8 {\rm GeV}$. This implies that calculations of the inelastic cross section that use current parton distribution functions and lowest order weak perturbation theory are unreliable above this energy.Comment: 11 pages, 3 figures, RevTeX, additional reference

Fate of lesion-related side branches after coronary artery stenting

AbstractObjectives. The aim of this study was to assess the immediate and long-term patency of lesion-associated side branches after coronary artery stenting.Background. The possible adverse effects related to implantation of coronary stents are not completely known. An important potential complication of stenting is side branch occlusion due to mechanical obstruction or thrombosis.Methods. Serial coronary angiography was performed in 153 patients (167 lesions) at baseline, after conventional balloon angioplasty, immediately after Palmaz-Schatz stent placement and at 6 months. The patency of side branches, where present, was analysed at each of these points.Results. Of 167 lesions stented, 57 stent placements spanned 66 side branches with a diameter ≥1 mm. Twenty-seven (41%) of these side branches had ≥50% ostial stenosis before standard balloon angioplasty. Six side branches became occluded after standard balloon angioplasty and remained occluded after stenting. Of the 60 side branches patent after conventional angioplasty, 57 (95%) remained patent immediately after stenting. All three side branches that became occluded after stenting had ≥50% ostial stenosis at baseline. All 60 side branches, including the 3 initially occluded after stenting, were patent at 6-month follow-up.Conclusions. These findings demonstrate that 1) acute side branch occlusion due to coronary stenting occurs infrequently; 2) when side branch occlusion occurs, it is associated with intrinsic ostial disease; and 3) the patency of side branch ostia is well maintained at long-term follow-up