A Budding-Defective M2 Mutant Exhibits Reduced Membrane Interaction, Insensitivity To Cholesterol, And Perturbed Interdomain Coupling

Influenza A M2 is a membrane-associated protein with a C-terminal amphipathic helix that plays a cholesterol-dependent role in viral budding. An M2 mutant with alanine substitutions in the C-terminal amphipathic helix is deficient in viral scission. With the goal of providing atomic-level understanding of how the wild-type protein functions, we used a multipronged site-directed spin labeling electron paramagnetic resonance spectroscopy (SDSL-EPR) approach to characterize the conformational properties of the alanine mutant. We spin-labeled sites in the transmembrane (TM) domain and the C-terminal amphipathic helix (AH) of wild-type (WT) and mutant M2, and collected information on line shapes, relaxation rates, membrane topology, and distances within the homotetramer in membranes with and without cholesterol. Our results identify marked differences in the conformation and dynamics between the WT and the alanine mutant. Compared to WT, the dominant population of the mutant AH is more dynamic, shallower in the membrane, and has altered quaternary arrangement of the C-terminal domain. While the AH becomes more dynamic, the dominant population of the TM domain of the mutant is immobilized. The presence of cholesterol changes the conformation and dynamics of the WT protein, while the alanine mutant is insensitive to cholesterol. These findings provide new insight into how M2 may facilitate budding. We propose the AH–membrane interaction modulates the arrangement of the TM helices, effectively stabilizing a conformational state that enables M2 to facilitate viral budding. Antagonizing the properties of the AH that enable interdomain coupling within M2 may therefore present a novel strategy for anti-influenza drug design

A Titanium Nitride Absorber for Controlling Optical Crosstalk in Horn-Coupled Aluminum LEKID Arrays for Millimeter Wavelengths

We discuss the design and measured performance of a titanium nitride (TiN) mesh absorber we are developing for controlling optical crosstalk in horn-coupled lumped-element kinetic inductance detector arrays for millimeter-wavelengths. This absorber was added to the fused silica anti-reflection coating attached to previously-characterized, 20-element prototype arrays of LEKIDs fabricated from thin-film aluminum on silicon substrates. To test the TiN crosstalk absorber, we compared the measured response and noise properties of LEKID arrays with and without the TiN mesh. For this test, the LEKIDs were illuminated with an adjustable, incoherent electronic millimeter-wave source. Our measurements show that the optical crosstalk in the LEKID array with the TiN absorber is reduced by 66\% on average, so the approach is effective and a viable candidate for future kilo-pixel arrays.Comment: 7 pages, 5 figures, accepted for publication in the Journal of Low Temperature Physic

Horn-Coupled, Commercially-Fabricated Aluminum Lumped-Element Kinetic Inductance Detectors for Millimeter Wavelengths

We discuss the design, fabrication, and testing of prototype horn-coupled, lumped-element kinetic inductance detectors (LEKIDs) designed for cosmic microwave background (CMB) studies. The LEKIDs are made from a thin aluminum film deposited on a silicon wafer and patterned using standard photolithographic techniques at STAR Cryoelectronics, a commercial device foundry. We fabricated twenty-element arrays, optimized for a spectral band centered on 150 GHz, to test the sensitivity and yield of the devices as well as the multiplexing scheme. We characterized the detectors in two configurations. First, the detectors were tested in a dark environment with the horn apertures covered, and second, the horn apertures were pointed towards a beam-filling cryogenic blackbody load. These tests show that the multiplexing scheme is robust and scalable, the yield across multiple LEKID arrays is 91%, and the noise-equivalent temperatures (NET) for a 4 K optical load are in the range 26\thinspace\pm6 \thinspace \mu \mbox{K} \sqrt{\mbox{s}}

The Detector System for the Stratospheric Kinetic Inductance Polarimeter (SKIP)

The Stratospheric Kinetic Inductance Polarimeter (SKIP) is a proposed balloon-borne experiment designed to study the cosmic microwave background, the cosmic infrared background and Galactic dust emission by observing 1133 square degrees of sky in the Northern Hemisphere with launches from Kiruna, Sweden. The instrument contains 2317 single-polarization, horn-coupled, aluminum lumped-element kinetic inductance detectors (LEKID). The LEKIDs will be maintained at 100 mK with an adiabatic demagnetization refrigerator. The polarimeter operates in two configurations, one sensitive to a spectral band centered on 150 GHz and the other sensitive to 260 and 350 GHz bands. The detector readout system is based on the ROACH-1 board, and the detectors will be biased below 300 MHz. The detector array is fed by an F/2.4 crossed-Dragone telescope with a 500 mm aperture yielding a 15 arcmin FWHM beam at 150 GHz. To minimize detector loading and maximize sensitivity, the entire optical system will be cooled to 1 K. Linearly polarized sky signals will be modulated with a metal-mesh half-wave plate that is mounted at the telescope aperture and rotated by a superconducting magnetic bearing. The observation program consists of at least two, five-day flights beginning with the 150 GHz observations.Comment: J Low Temp Phys DOI 10.1007/s10909-013-1014-3 The final publication is available at link.springer.co

A LEKID-based CMB instrument design for large-scale observations in Greenland

We present the results of a feasibility study, which examined deployment of a ground-based millimeter-wave polarimeter, tailored for observing the cosmic microwave background (CMB), to Isi Station in Greenland. The instrument for this study is based on lumped-element kinetic inductance detectors (LEKIDs) and an F/2.4 catoptric, crossed-Dragone telescope with a 500 mm aperture. The telescope is mounted inside the receiver and cooled to $<\,4$ K by a closed-cycle $^4$He refrigerator to reduce background loading on the detectors. Linearly polarized signals from the sky are modulated with a metal-mesh half-wave plate that is rotated at the aperture stop of the telescope with a hollow-shaft motor based on a superconducting magnetic bearing. The modular detector array design includes at least 2300 LEKIDs, and it can be configured for spectral bands centered on 150~GHz or greater. Our study considered configurations for observing in spectral bands centered on 150, 210 and 267~GHz. The entire polarimeter is mounted on a commercial precision rotary air bearing, which allows fast azimuth scan speeds with negligible vibration and mechanical wear over time. A slip ring provides power to the instrument, enabling circular scans (360 degrees of continuous rotation). This mount, when combined with sky rotation and the latitude of the observation site, produces a hypotrochoid scan pattern, which yields excellent cross-linking and enables 34\% of the sky to be observed using a range of constant elevation scans. This scan pattern and sky coverage combined with the beam size (15~arcmin at 150~GHz) makes the instrument sensitive to $5 < \ell < 1000$ in the angular power spectra

Welding fume nanoparticles from solid and flux-cored wires: Solubility, toxicity, and role of fluorides

Welding fume particles are hazardous. Their toxicity likely depends on their composition and reactivity. This study aimed at exploring the role of sodium or other fluorides (NaF), which are intentionally added to flux-cored wire electrodes for stainless steel welding, on the solubility (in phosphate buffered saline) and toxicity of the generated welding fume particles. A multi-analytical particle characterization approach along with in-vitro cell assays was undertaken. The release of Cr(VI) and Mn from the particles was tested as a function of fluoride solution concentration. The welding fume particles containing NaF released significantly higher amounts of Cr(VI) compared with solid wire reference fumes, which was associated with increased cytotoxicity and genotoxicity in-vitro. No crystalline Na or potassium (K) containing chromates were observed. Cr(VI) was incorporated in an amorphous mixed oxide. Solution-added fluorides did not increase the solubility of Cr(VI), but contributed to a reduced Mn release from both solid and flux-cored wire fume particles and the reduction of Cr(VI) release from solid wire fume particles. Chemical speciation modeling suggested that metal fluoride complexes were not formed. The presence of NaF in the welding electrodes did not have any direct, but possibly an indirect, role in the Cr(VI) solubility of welding fumes