Decay rate measurement of the first vibrationally excited state of MgH+^+ in a cryogenic Paul trap

We present a method to measure the decay rate of the first excited vibrational state of simple polar molecular ions being part of a Coulomb crystal in a cryogenic linear Paul trap. Specifically, we have monitored the decay of the $|\nu$=$1,J$=$1 \rangle_X$ towards the $|\nu$=$0,J$=$0 \rangle_X$ level in MgH$^+$ by saturated laser excitation of the $|\nu$=$0,J$=$2 \rangle_X$-$|\nu$=$1,J$=$1 \rangle_X$ transition followed by state selective resonance enhanced two-photon dissociation out of the $|\nu$=$0,J$=$2 \rangle_X$ level. The technique enables the determination of decay rates, and thus absorption strengths, with an accuracy at the few percent level.Comment: 5 pages, 4 figure

Imaging of cell membrane topography using Tamm plasmon coupled emission

Imaging of the cell membrane topography is important for a clear understanding of various biological activities of cells. We propose a technique for imaging the cell membrane topography that uses a metal-photonic crystal structure instead of a glass-water interface used in conventional polarized total internal reflection fluorescence microscopy (pTIRFM) techniques. Through the metal-photonic crystal of the proposed technique, the fluorophore labels on the cell membrane can be excited by both the p- and s-polarized excitation light, and in each case, the p- and s-polarized radiation from the excited fluorophores can be separated to form an image. We calculate the images of the cell membrane topography that is fusing a granule using the proposed technique and pTIRFM. The image obtained by the proposed technique shows a much greater contrast with respect to the background than that of the image obtained by pTIRFM. We also find that the structural similarity index of the image obtained by the proposed technique to a reference image is ~77%, which is only ~16% for the image obtained by pTIRFM. The proposed technique will help to obtain a clearer and more accurate image of the cell membrane topography, and hence, a deeper understanding of different biological activities

The unacknowledged legacy

This paper presents a critical discussion of the treatment of mimetic art, and particularly poetry and the theatre, in the work of the Athenian philosopher Plato (427-347 BC). It centres on Plato's discussion of the corrupting powers of the arts in the Republic, and the implications that his fierce attack on poetry and theatre have for his construction of the ideal polity. The legacy of Platonic ideas in later elaborations of the corrupting power of the arts is discussed. Furthermore, the paper investigates the relationship between current debates on cultural policy and the Platonic idea that the transformative powers of the arts ought to be harnessed by the state to promote a just society. The conclusion thus reached is that “instrumental cultural policy”, rather then being a modern invention, was in fact first theorized precisely in Plato's Republic

Photoaffinity cross-linking and unnatural amino acid mutagenesis reveal insights into calcitonin gene-related peptide binding to the calcitonin receptor-like receptor/receptor activity-modifying protein 1 (CLR/RAMP1) complex

Calcitonin gene-related peptide (CGRP) binds to the complex of the calcitonin receptor-like receptor (CLR) with receptor activity-modifying protein 1 (RAMP1). How CGRP interacts with the transmembrane domain (including the extracellular loops) of this family B receptor remains unclear. In this study, a photoaffinity cross-linker, p-azido l-phenylalanine (azF), was incorporated into CLR, chiefly in the second extracellular loop (ECL2) using genetic code expansion and unnatural amino acid mutagenesis. The method was optimized to ensure efficient photolysis of azF residues near the transmembrane bundle of the receptor. A CGRP analogue modified with fluorescein at position 15 was used for detection of ultraviolet-induced cross-linking. The methodology was verified by confirming the known contacts of CGRP to the extracellular domain of CLR. Within ECL2, the chief contacts were I284 on the loop itself and L291, at the top of the fifth transmembrane helix (TM5). Minor contacts were noted along the lip of ECL2 between S286 and L290 and also with M223 in TM3 and F349 in TM6. Full length molecular models of the bound receptor complex suggest that CGRP sits at the top of the TM bundle, with Thr6 of the peptide making contacts with L291 and H295. I284 is likely to contact Leu12 and Ala13 of CGRP, and Leu16 of CGRP is at the ECL/extracellular domain boundary of CLR. The reduced potency, Emax, and affinity of [Leu16Ala]-human α CGRP are consistent with this model. Contacts between Thr6 of CGRP and H295 may be particularly important for receptor activation

Estimation of the prevalence of cholesteryl ester storage disorder in a cohort of patients with clinical features of familial hypercholesterolaemia

Background and aim Familial hypercholesterolaemia is caused by variants in the low-density lipoprotein cholesterol metabolic pathway involving LDLR, APOB and PCSK9 genes. A national genetic testing service in Wales, UK has observed that no familial hypercholesterolaemia variant is found in almost 80% patients with the familial hypercholesterolaemia phenotype. It has recently been suggested that some adult patients with a familial hypercholesterolaemia phenotype may have cholesteryl ester storage disease which can also present as a mixed hyperlipidaemia. The commonest genetic cause of cholesteryl ester storage disease is an exon 8 splice junction variant in the LIPA gene (rs116928232, c.894G>A; E8SJM) previously found to have an allele frequency of 0.0011 (1 in 450 individuals) in a large European population. This study investigated the prevalence of the E8SJM in patients with a familial hypercholesterolaemia phenotype in Wales, UK. Method A total of 1203 patients with a clinical suspicion of familial hypercholesterolaemia but no familial hypercholesterolaemia variant were invited to participate. Of these, 668 patients provided informed written consent. Stored DNA samples from 663 patients were genotyped for the E8SJM variant. Results Three heterozygotes were identified (allele frequency 0.0023). Whole gene sequencing of the LIPA gene was undertaken in these three individuals, but no other variants were found. Therefore, there were no cholesteryl ester storage disease patients (homozygote or compound heterozygote) identified in this cohort. Conclusion The allele frequency 0.0023 (1 in 221 individuals) for the E8SJM variant was more prevalent in this cohort than in a European population study; however, no cholesteryl ester storage disease homozygotes were identified. We found no evidence to support routine testing for cholesteryl ester storage disease in adult patients with a familial hypercholesterolaemia phenotype

Glycomacropeptide:long-term use and impact on blood phenylalanine, growth and nutritional status in children with PKU

In phenylketonuria, casein glycomacropeptide (CGMP) requires modification with the addition of some essential and semi essential amino acids to ensure suitability as a protein substitute. The optimal amount and ratio of additional amino acids is undefined.AimA longitudinal, parallel, controlled study over 12months evaluating a CGMP (CGMP-AA2) formulation compared with phenylalanine-free L-amino acid supplements (L-AA) on blood Phe, Tyr, Phe:Tyr ratio, biochemical nutritional status and growth in children with PKU. The CGMP-AA2 contained 36mg Phe per 20g protein equivalent.MethodsChildren with PKU, with a median age of 9.2 y (5-16y) were divided into 2 groups: 29 were given CGMP-AA2, 19 remained on Phe-free L-AA. The CGMP-AA2 formula gradually replaced L-AA, providing blood Phe concentrations were maintained within target range. Median blood Phe, Tyr, Phe:Tyr ratio and anthropometry, were compared within and between the two groups at baseline, 26 and 52weeks. Nutritional biochemistry was studied at baseline and 26weeks only.ResultsAt the end of 52weeks only 48% of subjects were able to completely use CGMP-AA2 as their single source of protein substitute. At 52weeks CGMP-AA2 provided a median of 75% (30-100) of the total protein substitute with the remainder being given as L-AA. Within the CGMP-AA2 group, blood Phe increased significantly between baseline and 52weeks: [baseline to 26weeks; baseline Phe 270mol/L (170-430); 26weeks, Phe 300mol/L (125-485) p=0.06; baseline to 52weeks: baseline, Phe 270mol/L (170-430), 52weeks Phe 300mol/L (200-490),

Receptor activity-modifying proteins 2 and 3 generate adrenomedullin receptor subtypes with distinct molecular properties

Adrenomedullin (AM) is a peptide hormone with numerous effects in the vascular systems. AM signals through the AM1 and AM2 receptors formed by the obligate heterodimerization of a G protein-coupled receptor, the calcitonin receptor-like receptor (CLR), and receptor activity-modifying proteins (RAMP) 2 and 3, respectively. These different CLR-RAMP interactions yield discrete receptor pharmacology and physiological effects. The effective design of therapeutics that target the individual AM receptors is dependent on understanding the molecular details of the effects of RAMPs on CLR. To understand the role of RAMPs 2 and 3 on the activation and conformation of the CLR subunit of AM receptors we mutated 68 individual amino acids in the juxtamembrane region of CLR, a key region for activation of AM receptors and determined the effects on cAMP signalling. Sixteen CLR mutations had differential effects between the AM1 and AM2 receptors. Accompanying this, independent molecular modelling of the full-length AM-bound AM1 and AM2 receptors predicted differences in the binding pocket, and differences in the electrostatic potential of the two AM receptors. Druggability analysis indicated unique features that could be used to develop selective small molecule ligands for each receptor. The interaction of RAMP2 or RAMP3 with CLR induces conformational variation in the juxtamembrane region, yielding distinct binding pockets, probably via an allosteric mechanism. These subtype-specific differences have implications for the design of therapeutics aimed at specific AM receptors and for understanding the mechanisms by which accessory proteins affect G protein-coupled receptor function

Distinct Patterns of Internalization of Different Calcitonin GeneRelated Peptide Receptors

Calcitonin gene-related peptide (CGRP) is a neuropeptide that is involved in the transmission of pain. Drugs targeting CGRP or a CGRP receptor are efficacious in the treatment of migraine. The canonical CGRP receptor is a complex of a G protein-coupled receptor, the calcitonin-like receptor (CLR), with an accessory protein, receptor activity-modifying protein 1 (RAMP1). A second receptor, the AMY1 receptor, a complex of the calcitonin receptor with RAMP1, is a dual high-affinity receptor for CGRP and amylin. Receptor regulatory processes, such as internalization, are crucial for controlling peptide and drug responsiveness. Given the importance of CGRP receptor activity in migraine we compared the internalization profiles of both receptors for CGRP using novel fluorescent probes and a combination of live cell imaging, fixed cell imaging, and ELISA. This revealed stark differences in the regulation of each receptor with the AMY1 receptor unexpectedly showing little internalization.Peer Reviewe

THE THREE-DIMENSIONAL EVOLUTION OF ION-SCALE CURRENT SHEETS: TEARING AND DRIFT-KINK INSTABILITIES IN THE PRESENCE OF PROTON TEMPERATURE ANISOTROPY

We present the first three-dimensional hybrid simulations of the evolution of ion-scale current sheets, with an investigation of the role of temperature anisotropy and associated kinetic instabilities on the growth of the tearing instability and particle heating. We confirm the ability of the ion cyclotron and firehose instabilities to enhance or suppress reconnection, respectively. The simulations demonstrate the emergence of persistent three-dimensional structures, including patchy reconnection sites and the fast growth of a narrow-band drift-kink instability, which suppresses reconnection for thin current sheets with weak guide fields. Potential observational signatures of the three-dimensional evolution of solar wind current sheets are also discussed. We conclude that kinetic instabilities, arising from non-Maxwellian ion populations, are significant to the evolution of three-dimensional current sheets, and two-dimensional studies of heating rates by reconnection may therefore over-estimate the ability of thin, ion-scale current sheets to heat the solar wind by reconnection