Two Amino Acid Residues Contribute to a Cation-π Binding Interaction in the Binding Site of an Insect GABA Receptor

Cys-loop receptor binding sites characteristically possess an "aromatic box," where several aromatic amino acid residues surround the bound ligand. A cation-π interaction between one of these residues and the natural agonist is common, although the residue type and location are not conserved. Even in the closely related vertebrate GABA_A and GABA_C receptors, residues in distinct locations perform this role: in GABA_A receptors, a Tyr residue in loop A forms a cation-π interaction with GABA, while in GABA_C receptors it is a loop B residue. GABA-activated Cys-loop receptors also exist in invertebrates, where they have distinct pharmacologies and are the target of a range of pesticides. Here we examine the location of GABA in an insect binding site by incorporating a series of fluorinated Phe derivatives into the receptor binding pocket using unnatural amino acid mutagenesis, and evaluating the resulting receptors when expressed in Xenopus oocytes. A homology model suggests that two aromatic residues (in loops B and C) are positioned such that they could contribute to a cation-π interaction with the primary ammonium of GABA, and the data reveal a clear correlation between the GABA EC_(50) and the cation-π binding ability both at Phe206 (loop B) and Tyr254 (loop C), demonstrating for the first time the contribution of two aromatic residues to a cation-π interaction in a Cys-loop receptor

A Chromosomal Deletion and New Frameshift Mutation Cause ARSACS in an African-American

Autosomal Recessive Spastic Ataxia of Charlevoix-Saguenay (ARSACS) is a rare, progressive, neurodegenerative disease characterized by ataxia, spasticity and polyneuropathy. First described in the French-Canadian population of Quebec in 1978, ARSACS has since been identified in multiple patients worldwide. In this clinical case report, we describe the evaluation of an 11-years-old African-American male who presented to neuromuscular clinic for assessment of a gait abnormality. He had a history of gross motor delay since early childhood, frequent falls and a below average IQ. Chromosomal microarray revealed a 1.422 megabase loss in the 13q12.12 region, which includes the SACS gene. Next Generation Sequencing then showed a novel, predicted to be pathogenic missense mutation (c.11824dup) of this gene. His clinical presentation and neurological imaging further confirmed the diagnosis of ARSACS. To our knowledge, this is the first reported case of this disease in the African-American population of the United States. This case report further highlights the growing trend of identifying genetic diseases previously restricted to single, ethnically isolated regions in many different ethnic groups worldwide

ALMA observations of the supergiant B[e] star Wd1-9

Mass-loss in massive stars plays a critical role in their evolution, although the precise mechanism(s) responsible – radiatively driven winds, impulsive ejection and/or binary interaction – remain uncertain. In this Letter, we present Atacama Large Millimetre/Submillimeter Array line and continuum observations of the supergiant B[e] star Wd1-9, a massive post-main-sequence object located within the starburst cluster Westerlund 1 (Wd1). We find it to be one of the brightest stellar point sources in the sky at millimetre wavelengths, with (serendipitously identified) emission in the H41α radio recombination line. We attribute these properties to a low velocity (∼100 km s-1 ) ionized wind, with an extreme mass-loss rate ≳6.4 × 105(d/5 kpc)1.5 Mȯyr-1. External to this is an extended aspherical ejection nebula indicative of a prior phase of significant mass-loss. Taken together, the millimetre properties of Wd1-9 show a remarkable similarity to those of the highly luminous stellar source MWC349A. We conclude that these objects are interacting binaries evolving away from the main sequence and undergoing rapid case-A mass transfer. As such they – and by extension the wider class of supergiant B[e] stars – may provide a unique window into the physics of a process that shapes the life-cycle of ∼70 per cent of massive stars found in binary systems

Study of hose and connectors in life support systems Final report, 23 Feb. 1968 - 1 Mar. 1969

Nonflammable materials capable of fabrication into flexible oxygen hose for spacecraft life support syste

The Liquidus Temperature for Methanol-Water Mixtures at High Pressure and Low Temperature, with Application to Titan

Methanol is a potentially important impurity in subsurface oceans on Titan and Enceladus. We report measurements of the freezing of methanol-water samples at pressures up to 350~MPa using a volumetric cell with sapphire windows. For low concentrations of methanol, the liquidus temperature is typically a few degrees below the corresponding ice freezing point, while at high concentrations it follows the pure methanol trend. In the Ice-III regime, we observe several long-lived metastable states. The results suggest that methanol is a more effective antifreeze than previously estimated, and might have played an important role in the development of Titan's subsurface ocean

An Electrically Detonated Downhole Seismic Gun

An electrically detonated downhole seismic gun (EDG) that will fire blank 8-guage shells underwater has been constructed and tested to 80m depth (hydrostatic pressures of 130 psi or 8.9 x 105 Pa). Although other engineering seismic guns which fire blank or projectile sources are available, they are for near-surface shots and are not meant to be used for downhold seismic surveys in water-filled boreholes. The EDG was designed primarily for checkshot surveys (well travel-time surveys) and high-quality reflection/refraction tests, but potential applications include shallow vertical seismic profiling and borehole to borehole or borehole to surface tomography, as well as optimum offset and common midpoint seismic reflection surveys. The EDG consists off four steel subassemblies: (1) chamber; (2) breech; (3) pipe; and (4) hanger. A blank 8-gauge electrical shell is held by the chamber and is detonated by an electrode located inside the breech. O-ring seals prevent water from entering the breech and causing short circuits. The breech is screwed into a pipe which is also fitted with o-ring seals to keep the internal wiring dry. A hanger subassembly provides a convenient attaching point for the hoist cable. Arming of the EDG with an explosives blaster occurs only after lowering to operating depth. The EDG has been tested with various size black powder loads up to 750 grains. Frequency bandwidth and repeatability tests were carried out under saturated conditions in a fluvial and lacustrine sedimentary section. These preliminary tests show significant frequency content in the 25-200 Hz band (peak near 100 Hz) for reflections from depths of 150-300 m, and acceptable waveform repeatability for different shot records obtained with identical geometry and acquisition parameters

Structural Requirements in the Transmembrane Domain of GLIC Revealed by Incorporation of Noncanonical Histidine Analogs

The cyanobacterial pentameric ligand-gated ion channel GLIC, a homolog of the Cys-loop receptor superfamily, has provided useful structural and functional information about its eukaryotic counterparts. X-ray diffraction data and site-directed mutagenesis have previously implicated a transmembrane histidine residue (His234) as essential for channel function. Here, we investigated the role of His234 via synthesis and incorporation of histidine analogs and α-hydroxy acids using in vivo nonsense suppression. Receptors were expressed heterologously in Xenopus laevis oocytes, and whole-cell voltage-clamp electrophysiology was used to monitor channel activity. We show that an interhelix hydrogen bond involving His234 is important for stabilization of the open state, and that the shape and basicity of its side chain are highly sensitive to perturbations. In contrast, our data show that two other His residues are not involved in the acid-sensing mechanism

The Evaluation of the Effective Diameter (Deff) Calculation and Its Impact on the Size-Specific Dose Estimate (SSDE)

Information on the effective diameter (Deff) is essential for estimating the dose for patients undergoing CT examinations. The purpose of this study was to calculate the effective diameter using the maximum values of lateral (LAT) and anterior-posterior (AP) diameters (Deff,m) and using LAT and AP diameters taken from the center of the image (Deff,c), and compared both estimates to the effective diameter calculated directly from the cross-sectional area of the patient (Deff,A). We evaluated 164 patients who underwent the four most frequent CT examinations, namely pelvic, abdominal, thoracic, and head examinations, using a multi-detector CT (MDCT), the Toshiba Aquilion 128. We calculated the percentage differences between Deff,c and Deff,m from Deff,A. We also used the Wilcoxon-Mann-Witney U test to statistically determine whether differences were significant. While Deff,m is statistically no different (p > 0.05) from Deff,A, Deff,c is statistically different (p ˂ 0.05) from Deff,A except for head examinations.Received: 31 May 2016; Revised: 14 December 2016; Accepted: 20 January 201

A Cation-π Interaction in the Binding Site of the Glycine Receptor Is Mediated by a Phenylalanine Residue

Cys-loop receptor binding sites characteristically contain many aromatic amino acids. In nicotinic ACh and 5-HT3 receptors, a Trp residue forms a cation-{pi} interaction with the agonist, whereas in GABAA receptors, a Tyr performs this role. The glycine receptor binding site, however, contains predominantly Phe residues. Homology models suggest that two of these Phe side chains, Phe159 and Phe207, and possibly a third, Phe63, are positioned such that they could contribute to a cation-{pi} interaction with the primary amine of glycine. Here, we test this hypothesis by incorporation of a series of fluorinated Phe derivatives using unnatural amino acid mutagenesis. The data reveal a clear correlation between the glycine EC50 value and the cation-{pi} binding ability of the fluorinated Phe derivatives at position 159, but not at positions 207 or 63, indicating a single cation-{pi} interaction between glycine and Phe159. The data thus provide an anchor point for locating glycine in its binding site, and demonstrate for the first time a cation-{pi} interaction between Phe and a neurotransmitter