Structural and functional properties of ryanodine receptor type 3 in zebrafish tail muscle

The ryanodine receptor (RyR)1 isoform of the sarcoplasmic reticulum (SR) Ca2++ release channel is an essential component of all skeletal muscle fibers. RyR1s are detectable as "junctional feet" (JF) in the gap between the SR and the plasmalemma or T-tubules, and they are required for excitation-contraction (EC) coupling and differentiation. A second isoform, RyR3, does not sustain EC coupling and differentiation in the absence of RyR1 and is expressed at highly variable levels. Anatomically, RyR3 expression correlates with the presence of parajunctional feet (PJF), which are located on the sides of the SR junctional cisternae in an arrangement found only in fibers expressing RyR3. In frog muscle fibers, the presence of RyR3 and PJF correlates with the occurrence of Ca2++ sparks, which are elementary SR Ca2++ release events of the EC coupling machinery. Here, we explored the structural and functional roles of RyR3 by injecting zebrafish (Danio rerio) one-cell stage embryos with a morpholino designed to specifically silence RyR3 expression. In zebrafish larvae at 72 h postfertilization, fast-twitch fibers from wild-type (WT) tail muscles had abundant PJF. Silencing resulted in a drop of the PJF/JF ratio, from 0.79 in WT fibers to 0.03 in the morphants. The frequency with which Ca2++ sparks were detected dropped correspondingly, from 0.083 to 0.001 sarcomere-1 s-1. The few Ca2++ sparks detected in morphant fibers were smaller in amplitude, duration, and spatial extent compared with those in WT fibers. Despite the almost complete disappearance of PJF and Ca2++ sparks in morphant fibers, these fibers looked structurally normal and the swimming behavior of the larvae was not affected. This paper provides important evidence that RyR3 is the main constituent of the PJF and is the main contributor to the SR Ca2++ flux underlying Ca2++ sparks detected in fully differentiated frog and fish fibers. © 2015 Perni et al

Dynamics of the G-excess illusion

The G-excess illusion is increasingly recognized as a cause of aviation mishaps especially when pilots perform high-speed, steeply banked turns at low altitudes. Centrifuge studies of this illusion have examined the perception of subject orientation and/or target displacement during maintained hypergravity with the subject's head held stationary. The transient illusory perceptions produced by moving the head in hypergravity are difficult to study onboard centrifuges because the high angular velocity ensures the presence of strong Coriolis cross-coupled semicircular canal effects that mask immediate transient otolith-organ effects. The present study reports perceptions following head movements in hypergravity produced by high-speed aircraft maintaining a banked attitude with low angular velocity to minimize cross-coupled effects. Methods: Fourteen subjects flew on the NASA KC-135 and were exposed to resultant gravity forces of 1.3, 1.5, and 1.8 G for 3 minute periods. On command, seated subjects made controlled head movements in roll, pitch, and yaw at 30 second intervals both in the dark and with faint targets at a distance of 5 feet. Results: head movement produced transient perception of target displacement and velocity at levels as low as 1.3 G. Reports of target velocity without appropriate corresponding displacement were common. At 1.8 G when yaw head movements were made from a face down position, 4 subjects reported oscillatory rotational target displacement with fast and slow alternating components suggestive of torsional nystagmus. Head movements evoked symptoms of nausea in most subjects, with 2 subjects and 1 observer vomiting. Conclusions: The transient percepts present conflicting signals, which introduced confusion in target and subject orientation. Repeated head movements in hypergravity generate nausea by mechanisms distinct from cross-coupled Coriolis effects

Development of innovative fuelling systems for fusion energy science

The development of innovative fueling systems in support of magnetic fusion energy, particularly the International Thermonuclear Experimental Reactor (ITER), is described. The ITER fuelling system will use a combination of deuterium-tritium (D-T) gas puffing and pellet injection to achieve and maintain ignited plasmas. This combination will provide a flexible fuelling source with D-T pellets penetrating beyond the separatrix to sustain the ignited fusion plasma and with deuterium-rich gas fuelling the edge region to meet divertor requirements in a process called isotopic fuelling. More advanced systems with potential for deeper penetration, such as multistage pellet guns and compact toroid injection, are also described

Extraction of poloidal velocity from charge exchange recombination spectroscopy measurements

A novel approach has been implemented on DIII-D to allow the correct determination of the plasma poloidal velocity from charge exchange spectroscopy measurements. Unlike usual techniques, the need for detailed atomic physics calculations to properly interpret the results is alleviated. Instead, the needed atomic physics corrections are self-consistently determined directly from the measurements, by making use of specially chosen viewing chords. Modeling results are presented that were used to determine a set of views capable of measuring the correction terms. We present the analysis of a quiescent H-mode discharge, illustrating that significant modifications to the velocity profiles are required in these high ion temperature conditions. We also present preliminary measurements providing the first direct comparison of the standard cross-section correction to the atomic physics calculations

Chromophore switch from 11-cis-dehydroretinal (A2) to 11-cis-retinal (A1) decreases dark noise in salamander red rods

Dark noise, light-induced noise and responses to brief flashes of light were recorded in the membrane current of isolated rods from larval tiger salamander retina before and after bleaching most of the native visual pigment, which mainly has the 11-cis-3,4-dehydroretinal (A2) chromophore, and regenerating with the 11-cis-retinal (A1) chromophore in the same isolated rods. The purpose was to test the hypothesis that blue-shifting the pigment by switching from A2 to A1 will decrease the rate of spontaneous thermal activations and thus intrinsic light-like noise in the rod. Complete recordings were obtained in five cells (21◦C). Based on the wavelength of maximum absorbance, λmax,A1 =502 nm and λmax,A2 =528 nm, the average A2 : A1 ratio determined from rod spectral sensitivities and absorbances was ∼0.74 : 0.26 in the native state and ∼0.09 : 0.91 in the final state. In the native (A2) state, the single-quantum response (SQR) had an amplitude of 0.41±0.03 pA and an integration time of 3.16±0.15 s (mean±S.E.M.). The low-frequency branch of the dark noise power spectrum was consistent with discrete SQR-like events occurring at a rate of 0.238±0.026 rod−1 s−1. The corresponding values in the final state were 0.57±0.07 pA (SQR amplitude), 3.47±0.26 s (SQR integration time), and 0.030±0.006 rod−1 s−1 (rate of dark events). Thus the rate of dark events per rod and the fraction of A2 pigment both changed by ca 8-fold between the native and final states, indicating that the dark events originated mainly in A2 molecules even in the final state. By extrapolating the linear relation between event rates and A2 fraction to 0% A2 (100% A1) and 100% A2 (0% A1), we estimated that the A1 pigment is at least 36 times more stable than the A2 pigment. The noise component attributed to discrete dark events accounted for 73% of the total dark current variance in the native (A2) state and 46% in the final state. The power spectrum of the remaining ‘continuous’ noise component did not differ between the two states. The smaller and faster SQR in the native (A2) state is consistent with the idea that the rod behaves as if light-adapted by dark events that occur at a rate of nearly one per integration time. Both the decreased level of dark noise and the increased SQR amplitude must significantly improve the reliability of photon detection in dim light in the presence of the A1 chromophore compared to the native (A2) state in salamander rods

New Pellet Injection Schemes on DIII-D

The pellet fueling system on DIII-D has been modified for injection of deuterium pellets from two vertical ports and two inner wall locations on the magnetic high-field side (HFS) of the tokamak. The HFS pellet injection technique was first employed on ASDEX-Upgrade with significant improvements reported in both pellet penetration and fueling efficiency. The new pellet injection schemes on DIII-D required the installation of new guide tubes. These lines are {approx_equal}12.5 m in total length and are made up of complex bends and turns (''roller coaster'' like) to route pellets from the injector to the plasma, including sections inside the torus. The pellet speed at which intact pellets can survive through the curved guide tubes is limited ({approx_equal}200-300 m/s for HFS injection schemes). Thus, one of the three gas guns on the injector was modified to provide pellets in a lower speed regime than the original guns (normal speed range {approx_equal}500 to 1000 m/s). The guide tube installations and gun modifications are described along with the injector operating parameters, and the latest test results are highlighted

A high-resolution map of human evolutionary constraint using 29 mammals.

The comparison of related genomes has emerged as a powerful lens for genome interpretation. Here we report the sequencing and comparative analysis of 29 eutherian genomes. We confirm that at least 5.5% of the human genome has undergone purifying selection, and locate constrained elements covering ∼4.2% of the genome. We use evolutionary signatures and comparisons with experimental data sets to suggest candidate functions for ∼60% of constrained bases. These elements reveal a small number of new coding exons, candidate stop codon readthrough events and over 10,000 regions of overlapping synonymous constraint within protein-coding exons. We find 220 candidate RNA structural families, and nearly a million elements overlapping potential promoter, enhancer and insulator regions. We report specific amino acid residues that have undergone positive selection, 280,000 non-coding elements exapted from mobile elements and more than 1,000 primate- and human-accelerated elements. Overlap with disease-associated variants indicates that our findings will be relevant for studies of human biology, health and disease

Human Wavelength Discrimination of Monochromatic Light Explained by Optimal Wavelength Decoding of Light of Unknown Intensity

We show that human ability to discriminate the wavelength of monochromatic light can be understood as maximum likelihood decoding of the cone absorptions, with a signal processing efficiency that is independent of the wavelength. This work is built on the framework of ideal observer analysis of visual discrimination used in many previous works. A distinctive aspect of our work is that we highlight a perceptual confound that observers should confuse a change in input light wavelength with a change in input intensity. Hence a simple ideal observer model which assumes that an observer has a full knowledge of input intensity should over-estimate human ability in discriminating wavelengths of two inputs of unequal intensity. This confound also makes it difficult to consistently measure human ability in wavelength discrimination by asking observers to distinguish two input colors while matching their brightness. We argue that the best experimental method for reliable measurement of discrimination thresholds is the one of Pokorny and Smith, in which observers only need to distinguish two inputs, regardless of whether they differ in hue or brightness. We mathematically formulate wavelength discrimination under this wavelength-intensity confound and show a good agreement between our theoretical prediction and the behavioral data. Our analysis explains why the discrimination threshold varies with the input wavelength, and shows how sensitively the threshold depends on the relative densities of the three types of cones in the retina (and in particular predict discriminations in dichromats). Our mathematical formulation and solution can be applied to general problems of sensory discrimination when there is a perceptual confound from other sensory feature dimensions