Actions of Agonists, Fipronil and Ivermectin on the Predominant In Vivo Splice and Edit Variant (RDLbd, I/V) of the Drosophila GABA Receptor Expressed in Xenopus laevis Oocytes

Ionotropic GABA receptors are the targets for several classes of insecticides. One of the most widely-studied insect GABA receptors is RDL (resistance to dieldrin), originally isolated from Drosophila melanogaster. RDL undergoes alternative splicing and RNA editing, which influence the potency of GABA. Most work has focussed on minority isoforms. Here, we report the first characterisation of the predominant native splice variant and RNA edit, combining functional characterisation with molecular modelling of the agonist-binding region. The relative order of agonist potency is GABA> muscimol> TACA> β-alanine. The I/V edit does not alter the potency of GABA compared to RDLbd. Docking calculations suggest that these agonists bind and activate RDLbdI/V through a similar binding mode. TACA and β-alanine are predicted to bind with lower affinity than GABA, potentially explaining their lower potency, whereas the lower potency of muscimol and isoguvacine cannot be explained structurally from the docking calculations. The A301S (resistance to dieldrin) mutation reduced the potency of antagonists picrotoxin, fipronil and pyrafluprole but the I/V edit had no measurable effect. Ivermectin suppressed responses to GABA of RDLbdI/V, RDLbd and RDLbdI/VA301S. The dieldrin resistant variant also showed reduced sensitivity to Ivermectin. This study of a highly abundant insect GABA receptor isoform will help the design of new insecticides

Dynamo constraints on the long-term evolution of Earth's magnetic field strength

Elucidating the processes in the liquid core that have produced observed palaeointensity changes over the last 3.5 Gyr is crucial for understanding the dynamics and long-term evolution of Earth’s deep interior. We combine numerical geodynamo simulations with theoretical scaling laws to investigate the variation of Earth’s magnetic field strength over geological time. Our approach follows the study of Aubert et al., adapted to include recent advances in numerical simulations, mineral physics and palaeomagnetism. We first compare the field strength within the dynamo region and on the core–mantle boundary (CMB) between a suite of 314 dynamo simulations and two power-based theoretical scaling laws. The scaling laws are both based on a Quasi-Geostropic (QG) force balance at leading order and a Magnetic, Archimedian, and Coriolis (MAC) balance at first order and differ in treating the characteristic length scale of the convection as fixed (QG-MAC-fixed) or determined as part of the solution (QG-MAC-free). When the data set is filtered to retain only simulations with magnetic to kinetic energy ratios greater than at least two we find that the internal field together with the root-mean-square and dipole CMB fields exhibit power-law behaviour that is compatible with both scalings within uncertainties arising from different heating modes and boundary conditions. However, while the extrapolated intensity based on the QG-MAC-free scaling matches Earth’s modern CMB field, the QG-MAC-fixed prediction shoots too high and also significantly overestimates palaeointensities over the last 3.5 Gyr. We combine the QG-MAC-free scaling with outputs from 275 realizations of core–mantle thermal evolution to construct synthetic true dipole moment (TDM) curves spanning the last 3.5 Gyr. Best-fitting TDMs reproduce binned PINT data during the Bruhnes and before inner core nucleation (ICN) within observational uncertainties, but PINT does not contain the predicted strong increase and subsequent high TDMs during the early stages of inner core growth. The best-fitting models are obtained for a present-day CMB heat flow of 11–16 TW, increasing to 17–22 TW at 4 Ga, and predict a minimum TDM at ICN

Utility of the FebriDx point-of-care assay in supporting a triage algorithm for medical admissions with possible COVID-19: an observational cohort study

Objective: To evaluate a triage algorithm used to identify and isolate patients with suspected COVID-19 among medical patients needing admission to hospital using simple clinical criteria and the FebriDx assay. Design:: Retrospective observational cohort. Setting Large acute National Health Service hospital in London, UK. Participants: All medical admissions from the emergency department between 10 August 2020 and 4 November 2020 with a valid SARS-CoV-2 RT-PCR result. Interventions: Medical admissions were triaged as likely, possible or unlikely COVID-19 based on clinical criteria. Patients triaged as possible COVID-19 underwent FebriDx lateral flow assay on capillary blood, and those positive for myxovirus resistance protein A (a host response protein) were managed as likely COVID-19. Primary outcome measures: Diagnostic accuracy (sensitivity, specificity and predictive values) of the algorithm and the FebriDx assay using SARS-CoV-2 RT-PCR from nasopharyngeal swabs as the reference standard. Results: 4.0% (136) of 3443 medical admissions had RT-PCR confirmed COVID-19. Prevalence of COVID-19 was 46% (80/175) in those triaged as likely, 4.1% (50/1225) in possible and 0.3% (6/2033) in unlikely COVID-19. Using a SARS-CoV-2 RT-PCR reference standard, clinical triage had sensitivity of 96% (95% CI 91% to 98%) and specificity of 61.5% (95% CI 59.8% to 63.1%), while the triage algorithm including FebriDx had sensitivity of 93% (95% CI 87% to 96%) and specificity of 86.4% (95% CI 85.2% to 87.5%). While 2033 patients were deemed not to require isolation using clinical criteria alone, the addition of FebriDx to clinical triage allowed a further 826 patients to be released from isolation, reducing the need for isolation rooms by 9.5 per day, 95% CI 8.9 to 10.2. Ten patients missed by the algorithm had mild or asymptomatic COVID-19. Conclusions: A triage algorithm including the FebriDx assay had good sensitivity and was useful to ‘rule-out’ COVID-19 among medical admissions to hospital

Full vector archaeomagnetic records from Anatolia between 2400 and 1350 BCE: Implications for geomagnetic field models and the dating of fires in antiquity

Anatolia, as one of the busiest crossroads of ancient civilizations, provides an ideal platform for archaeomagnetic studies. Previous results from the Middle East have suggested the occurrence of a strong peak in geomagnetic intensity at ∼1000 BCE associated with dramatic field strength variations that could require a radical rethinking of geodynamo theory. The behavior of the field in the centuries preceding this peak remains poorly constrained, however. Here we present the results of full-vector archaeomagnetic experiments performed on 18 sets of samples from three archaeological sites belonging to Assyrian Trade Colony and Hittite periods. Associated rock magnetic analyses showed that the major magnetic carrier is magnetite chemically stable up to 700 °C and the magnetic mineral assemblage is composed mostly of non-interacting PSD grains. The directional results are compared with existing data and with the most recent global geomagnetic field models pfm9k.1b and SHA.DIF.14k. The directions are in remarkably good agreement with SHA.DIF.14k which is based on archaeomagnetic and lava flow data. Together with our earlier results from Anatolia, we triple the existing database of directions for the 700 year long period 2200–1500 BCE, over a large region from Greece to Azerbaijan, and from Moldavia/Ukraine to Egypt. Three archaeointensity methods: thermal IZZI-Thellier, microwave Thellier and the multi-specimen protocol (MSP) produced virtual axial dipole moment estimates () that are somewhat higher than contemporaneous (regional and global) data and model predictions suggesting that the field was already substantially stronger than today more than 800 years prior to the reported peak. In addition to constraining geomagnetic variability, our data also allow us to assign relative dates to inferred fire events in the Assyrian Trade Colony Period sites. This allows us to conclude that the fire events at the largest site, Kültepe, were not all contemporaneous with one another and with the abandonment of the site as has been previously hypothesized

Paleoproterozoic Geomagnetic Field Strength From the Avanavero Mafic Sills, Amazonian Craton, Brazil

A recent hypothesis has suggested that Earth's inner core nucleated during the Mesoproterozoic, as evidenced by a rapid increase in the paleointensity (ancient geomagnetic field intensity) record; however, paleointensity data during the Paleoproterozoic and Mesoproterozoic period are limited. To address this problem, we have determined paleointensity from samples from three Paleoproterozoic Avanavero mafic sills (Amazonian Craton, Brazil): Cotingo, 1782 Ma, Puiuà 1788, and Pedra Preta, 1795 Ma. We adopted a multi-protocol approach for paleointensity estimates combining Thellier-type IZZI and LTD-IZZI methods, and the non-heating Preisach protocol. We obtained an average VDM value of 1.3 ± 0.7 × 1022Am2 (Cotingo) of 2.0 ± 0.4 × 1022Am2 (Puiuà) and 6 ± 4 × 1022Am2 (Pedra Preta); it is argued that the Cotingo estimate is the most robust. Our results are the first data from the upper Paleoproterozoic for South America and are comparable to data available from other regions and similar periods. The new data do not invalidate the hypothesis of that Earth's inner core nucleated during the Mesoproterozoic

Role of the Cys loop and transmembrane domain in the allosteric modulation of α4β2 nicotinic acetylcholine receptors

​Allosteric modulators of pentameric ligand gated ion channels (pLGICs) are thought to act on elements of the pathways that couple agonist binding to channel gating. Using α4β2 nicotinic acetylcholine receptors (nAChRs) and the α4β2-selective positive modulators 17β-estradiol (βEST) and desformylflustrabromine (dFBr), we have identified pathways that link the binding sites for these modulators to the Cys loop, a region that is critical for channel gating in all pLGICs. Previous studies have shown that the binding site for potentiating βEST is in the C-terminal (post-M4 region) of the α4 subunit. Here, using homology modelling in combination with mutagenesis and electrophysiology, we identified the binding site for potentiating dFBr on the top-half of a cavity between the third (M3) and fourth transmembrane (M4) α-helices of the α4 subunit. We found that the binding sites for βEST and dFBr communicate with the Cys loop, through interactions between the last residue of post-M4 and F170 of the conserved FPF sequence of the Cys loop, and that these interactions affect potentiating efficacy. In addition, interactions between a residue in M3 (Y309) and F167, a residue adjacent to the Cys loop FPF motif, also affect dFBr potentiating efficacy. Thus, the Cys loop acts as a key control element in the allosteric transduction pathway for potentiating βEST and dFBr. Overall, we propose that positive allosteric modulators that bind the M3-M4 cavity or post-M4 region increase the efficacy of channel gating through interactions with the Cys loop

Magnetic resonance imaging pattern recognition in childhood bilateral basal ganglia disorders

Bilateral basal ganglia abnormalities on MRI are observed in a wide variety of childhood disorders. MRI pattern recognition can enable rationalization of investigations and also complement clinical and molecular findings, particularly confirming genomic findings and also enabling new gene discovery. A pattern recognition approach in children with bilateral basal ganglia abnormalities on brain MRI was undertaken in this international multicentre cohort study. Three hundred and five MRI scans belonging to 201 children with 34 different disorders were rated using a standard radiological scoring proforma. In addition, literature review on MRI patterns was undertaken in these 34 disorders and 59 additional disorders reported with bilateral basal ganglia MRI abnormalities. Cluster analysis on first MRI findings from the study cohort grouped them into four clusters: Cluster 1—T2-weighted hyperintensities in the putamen; Cluster 2—T2-weighted hyperintensities or increased MRI susceptibility in the globus pallidus; Cluster 3—T2-weighted hyperintensities in the globus pallidus, brainstem and cerebellum with diffusion restriction; Cluster 4—T1-weighted hyperintensities in the basal ganglia. The 34 diagnostic categories included in this study showed dominant clustering in one of the above four clusters. Inflammatory disorders grouped together in Cluster 1. Mitochondrial and other neurometabolic disorders were distributed across clusters 1, 2 and 3, according to lesions dominantly affecting the striatum (Cluster 1: glutaric aciduria type 1, propionic acidaemia, 3-methylglutaconic aciduria with deafness, encephalopathy and Leigh-like syndrome and thiamine responsive basal ganglia disease associated with SLC19A3), pallidum (Cluster 2: methylmalonic acidaemia, Kearns Sayre syndrome, pyruvate dehydrogenase complex deficiency and succinic semialdehyde dehydrogenase deficiency) or pallidum, brainstem and cerebellum (Cluster 3: vigabatrin toxicity, Krabbe disease). The Cluster 4 pattern was exemplified by distinct T1-weighted hyperintensities in the basal ganglia and other brain regions in genetically determined hypermanganesemia due to SLC39A14 and SLC30A10. Within the clusters, distinctive basal ganglia MRI patterns were noted in acquired disorders such as cerebral palsy due to hypoxic ischaemic encephalopathy in full-term babies, kernicterus and vigabatrin toxicity and in rare genetic disorders such as 3-methylglutaconic aciduria with deafness, encephalopathy and Leigh-like syndrome, thiamine responsive basal ganglia disease, pantothenate kinase-associated neurodegeneration, TUBB4A and hypermanganesemia. Integrated findings from the study cohort and literature review were used to propose a diagnostic algorithm to approach bilateral basal ganglia abnormalities on MRI. After integrating clinical summaries and MRI findings from the literature review, we developed a prototypic decision-making electronic tool to be tested using further cohorts and clinical practice