Structural analysis of the carboxy terminal PH domain of pleckstrin bound to D-myo-inositol 1,2,3,5,6-pentakisphosphate

<p>Abstract</p> <p>Background</p> <p>Pleckstrin homology (PH) domains are one of the most prevalent domains in the human proteome and represent the major phosphoinositide-binding module. These domains are often found in signaling proteins and function predominately by targeting their host proteins to the cell membrane. Inositol phosphates, which are structurally similar to phosphoinositides, are not only known to play a role as signaling molecules but are also capable of being bound by PH domains.</p> <p>Results</p> <p>In the work presented here it is shown that the addition of commercial <it>myo</it>-inositol hexakisphosphate (IP₆) inhibited the binding of the carboxy terminal PH domain of pleckstrin (C-PH) to phosphatidylinositol 3,4-bisphosphate with an IC₅₀of 7.5 μM. In an attempt to characterize this binding structurally, C-PH was crystallized in the presence of IP₆and the structure was determined to 1.35 Å. Examination of the resulting electron density unexpectedly revealed the bound ligand to be D-<it>myo</it>-inositol 1,2,3,5,6-pentakisphosphate.</p> <p>Conclusion</p> <p>The discovery of D-<it>myo</it>-inositol 1,2,3,5,6-pentakisphosphate in the crystal structure suggests that the inhibitory effects observed in the binding studies may be due to this ligand rather than IP₆. Analysis of the protein-ligand interaction demonstrated that this <it>myo</it>-inositol pentakisphosphate isomer interacts specifically with protein residues known to be involved in phosphoinositide binding. In addition to this, a structural alignment of other PH domains bound to inositol phosphates containing either four or five phosphate groups revealed that the majority of phosphate groups occupy conserved locations in the binding pockets of PH domains. These findings, taken together with other recently reported studies suggest that <it>myo-</it>inositol pentakisphosphates could act to regulate PH domain-phosphoinositide interactions by directly competing for binding, thus playing an important role as signaling molecules.</p

Having a lot of a good thing: multiple important group memberships as a source of self-esteem.

Copyright: © 2015 Jetten et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are creditedMembership in important social groups can promote a positive identity. We propose and test an identity resource model in which personal self-esteem is boosted by membership in additional important social groups. Belonging to multiple important group memberships predicts personal self-esteem in children (Study 1a), older adults (Study 1b), and former residents of a homeless shelter (Study 1c). Study 2 shows that the effects of multiple important group memberships on personal self-esteem are not reducible to number of interpersonal ties. Studies 3a and 3b provide longitudinal evidence that multiple important group memberships predict personal self-esteem over time. Studies 4 and 5 show that collective self-esteem mediates this effect, suggesting that membership in multiple important groups boosts personal self-esteem because people take pride in, and derive meaning from, important group memberships. Discussion focuses on when and why important group memberships act as a social resource that fuels personal self-esteem.This study was supported by 1. Australian Research Council Future Fellowship (FT110100238) awarded to Jolanda Jetten (see http://www.arc.gov.au) 2. Australian Research Council Linkage Grant (LP110200437) to Jolanda Jetten and Genevieve Dingle (see http://www.arc.gov.au) 3. support from the Canadian Institute for Advanced Research Social Interactions, Identity and Well-Being Program to Nyla Branscombe, S. Alexander Haslam, and Catherine Haslam (see http://www.cifar.ca)

Structural characterisation of neutrophil glycans by ultra sensitive mass spectrometric glycomics methodology

Neutrophils are the most abundant white blood cells in humans and play a vital role in several aspects of the immune response. Numerous reports have implicated neutrophil glycosylation as an important factor in mediating these interactions. We report here the application of high sensitivity glycomics methodologies, including matrix assisted laser desorption ionisation (MALDI-TOF) and MALDI-TOF/TOF analyses, to the structural analysis of N- and O-linked carbohydrates released from two samples of neutrophils, prepared by two separate and geographically remote laboratories. The data produced demonstrates that the cells display a diverse range of sialylated and fucosylated complex glycans, with a high level of similarity between the two preparations

Coulomb drag propulsion experiments of ESTCube-2 and FORESAIL-1

This paper presents two technology experiments – the plasma brake for deorbiting and the electric solar wind sail for interplanetary propulsion – on board the ESTCube-2 and FORESAIL-1 satellites. Since both technologies employ the Coulomb interaction between a charged tether and a plasma flow, they are commonly referred to as Coulomb drag propulsion. The plasma brake operates in the ionosphere, where a negatively charged tether deorbits a satellite. The electric sail operates in the solar wind, where a positively charged tether propels a spacecraft, while an electron emitter removes trapped electrons. Both satellites will be launched in low Earth orbit carrying nearly identical Coulomb drag propulsion experiments, with the main difference being that ESTCube-2 has an electron emitter and it can operate in the positive mode. While solar-wind sailing is not possible in low Earth orbit, ESTCube-2 will space-qualify the components necessary for future electric sail experiments in its authentic environment. The plasma brake can be used on a range of satellite mass classes and orbits. On nanosatellites, the plasma brake is an enabler of deorbiting – a 300-m-long tether fits within half a cubesat unit, and, when charged with -1 kV, can deorbit a 4.5-kg satellite from between a 700- and 500-km altitude in approximately 9–13 months. This paper provides the design and detailed analysis of low-Earth-orbit experiments, as well as the overall mission design of ESTCube-2 and FORESAIL-1.Peer reviewe

Coulomb drag propulsion experiments of ESTCube-2 and FORESAIL-1

This paper presents two technology experiments – the plasma brake for deorbiting and the electric solar wind sail for interplanetary propulsion – on board the ESTCube-2 and FORESAIL-1 satellites. Since both technologies employ the Coulomb interaction between a charged tether and a plasma flow, they are commonly referred to as Coulomb drag propulsion. The plasma brake operates in the ionosphere, where a negatively charged tether deorbits a satellite. The electric sail operates in the solar wind, where a positively charged tether propels a spacecraft, while an electron emitter removes trapped electrons. Both satellites will be launched in low Earth orbit carrying nearly identical Coulomb drag propulsion experiments, with the main difference being that ESTCube-2 has an electron emitter and it can operate in the positive mode. While solar-wind sailing is not possible in low Earth orbit, ESTCube-2 will space-qualify the components necessary for future electric sail experiments in its authentic environment. The plasma brake can be used on a range of satellite mass classes and orbits. On nanosatellites, the plasma brake is an enabler of deorbiting – a 300-m-long tether fits within half a cubesat unit, and, when charged with - 1 kV, can deorbit a 4.5-kg satellite from between a 700- and 500-km altitude in approximately 9–13 months. This paper provides the design and detailed analysis of low-Earth-orbit experiments, as well as the overall mission design of ESTCube-2 and FORESAIL-1.</p

Multiorgan MRI findings after hospitalisation with COVID-19 in the UK (C-MORE): a prospective, multicentre, observational cohort study

Introduction: The multiorgan impact of moderate to severe coronavirus infections in the post-acute phase is still poorly understood. We aimed to evaluate the excess burden of multiorgan abnormalities after hospitalisation with COVID-19, evaluate their determinants, and explore associations with patient-related outcome measures. Methods: In a prospective, UK-wide, multicentre MRI follow-up study (C-MORE), adults (aged ≥18 years) discharged from hospital following COVID-19 who were included in Tier 2 of the Post-hospitalisation COVID-19 study (PHOSP-COVID) and contemporary controls with no evidence of previous COVID-19 (SARS-CoV-2 nucleocapsid antibody negative) underwent multiorgan MRI (lungs, heart, brain, liver, and kidneys) with quantitative and qualitative assessment of images and clinical adjudication when relevant. Individuals with end-stage renal failure or contraindications to MRI were excluded. Participants also underwent detailed recording of symptoms, and physiological and biochemical tests. The primary outcome was the excess burden of multiorgan abnormalities (two or more organs) relative to controls, with further adjustments for potential confounders. The C-MORE study is ongoing and is registered with ClinicalTrials.gov, NCT04510025. Findings: Of 2710 participants in Tier 2 of PHOSP-COVID, 531 were recruited across 13 UK-wide C-MORE sites. After exclusions, 259 C-MORE patients (mean age 57 years [SD 12]; 158 [61%] male and 101 [39%] female) who were discharged from hospital with PCR-confirmed or clinically diagnosed COVID-19 between March 1, 2020, and Nov 1, 2021, and 52 non-COVID-19 controls from the community (mean age 49 years [SD 14]; 30 [58%] male and 22 [42%] female) were included in the analysis. Patients were assessed at a median of 5·0 months (IQR 4·2–6·3) after hospital discharge. Compared with non-COVID-19 controls, patients were older, living with more obesity, and had more comorbidities. Multiorgan abnormalities on MRI were more frequent in patients than in controls (157 [61%] of 259 vs 14 [27%] of 52; p&lt;0·0001) and independently associated with COVID-19 status (odds ratio [OR] 2·9 [95% CI 1·5–5·8]; padjusted=0·0023) after adjusting for relevant confounders. Compared with controls, patients were more likely to have MRI evidence of lung abnormalities (p=0·0001; parenchymal abnormalities), brain abnormalities (p&lt;0·0001; more white matter hyperintensities and regional brain volume reduction), and kidney abnormalities (p=0·014; lower medullary T1 and loss of corticomedullary differentiation), whereas cardiac and liver MRI abnormalities were similar between patients and controls. Patients with multiorgan abnormalities were older (difference in mean age 7 years [95% CI 4–10]; mean age of 59·8 years [SD 11·7] with multiorgan abnormalities vs mean age of 52·8 years [11·9] without multiorgan abnormalities; p&lt;0·0001), more likely to have three or more comorbidities (OR 2·47 [1·32–4·82]; padjusted=0·0059), and more likely to have a more severe acute infection (acute CRP &gt;5mg/L, OR 3·55 [1·23–11·88]; padjusted=0·025) than those without multiorgan abnormalities. Presence of lung MRI abnormalities was associated with a two-fold higher risk of chest tightness, and multiorgan MRI abnormalities were associated with severe and very severe persistent physical and mental health impairment (PHOSP-COVID symptom clusters) after hospitalisation. Interpretation: After hospitalisation for COVID-19, people are at risk of multiorgan abnormalities in the medium term. Our findings emphasise the need for proactive multidisciplinary care pathways, with the potential for imaging to guide surveillance frequency and therapeutic stratification

Electric Sail and Plasma Brake Deployment System Prototype

This thesis will detail the prototyping and results of the main and peripheral functions of a deployment system for electric sail and plasma brake technologies. The aim of this project is to provide a fully functioning prototype that may be developed further into engineering qualification models, and finally culminating in a flight model. First the thesis will clarify cubesat technology, the science of these technologies, followed by covering the schematic and subsystems for the deployment system. The study utilises some FORESAIL-1 mission specifications as a basis for development. The prototype was implemented using standard retail components and a STM32 microcontroller embedded on an evaluation board. Programming of the STM32 was accomplished with Keil integrated development environment using C language. Command communication, for simulation of the commands an On-Board Computer would send to payloads, is achieved with YAT Terminal and a universal asynchronous receiver-transmitter interface. Measurements were executed with a digital multimeter, oscilloscope and the STM32s built-in analog-to-digital converters. For the results, the functionality of the prototype is analysed and assessed. The results are compared with the calculations. Finally this report will discuss the improvements that can be applied for future iterations.Thesis also consists of an electronic prototype realised on a printed circuit board

Forssman Synthetase Expression Results in Diminished Shiga Toxin Susceptibility: a Role for Glycolipids in Determining Host-Microbe Interactions

Forssman glycolipid (FG), the product of Forssman synthetase (FS), is widely expressed among nonprimate mammalian species. Here, we describe a molecular and genetic relationship between FG expression and Shiga toxin (Stx) susceptibility. We have isolated the FS cDNA from human, canine, and murine cells. Whereas the murine and canine FS genes express a functional enzyme, the human FS cDNA was found to express a protein that lacks FS activity, despite a high degree of sequence identity with the enzymatically active murine and canine FS genes. In order to examine the relationship between FG expression and Stx susceptibility, Vero cells were transfected with the three FS orthologues or a vector control. Complementation with the human FS cDNA had no effect on Stx susceptibility, whereas stable expression of the canine and murine FS resulted in markedly decreased susceptibility to toxin. Among individual cells, an inverse correlation between FG expression and Stx binding was demonstrated. Moreover, only strongly FG-reactive cells were capable of growing in the presence of Stx. These cells were found to have high levels of FG expression and a correspondingly diminished GbO(3) content. We conclude that expression of a functionally active FS modifies Stx receptor glycolipids to FG and results in markedly decreased susceptibility to toxin. We speculate that inactivation of the FS gene during primate evolution may account, at least in part, for the marked susceptibility of human cells to Stx

The socio-spatial politics of royalties and their distribution: A case study of the Surat Basin, Queensland

This paper is concerned with the socio-spatial and ethical politics of redistribution, specifically the allocation of natural resources rents from political and economic cores to the economic and geographical peripheries whence the resource originated. Based on a case study of the coal seam gas sector in Queensland's Surat Basin, this paper focuses on the operation of the Queensland State Government's regional development fund for mining and energy extraction-affected regions. Employing an environmental justice framework, we critically explore the operation of these funds in ostensibly helping constituent communities in becoming resilient to the worst effects of the 'staples trap'. Drawing on secondary demographic and housing data for the region, as well as primary information collected from key respondents from mid-2018 to early 2019, we show that funds were distributed across all of the local government areas, and allocated to projects and places primarily on a perceived economic needs basis. However, concerns were raised with the probity of the funds' administration. In terms of recognition justice, the participation of smaller and more remote towns and local Indigenous communities was hampered by their structural marginalisation. Procedurally, the funds were criticised for the lack of local consultation taken in the development and approval of projects. While spatially concentrated expenditure may be the most cost-effective use of public monies, we argue that grant application processes should be open, transparent and inclusive, and the outcomes cognisant of the developmental needs of smaller communities, together with the need to foster regional solidarity and coherence