Proton transfer pathway in anion channelrhodopsin-1

Anion channelrhodopsin from Guillardia theta (GtACR1) has Asp234 (3.2 angstrom) and Glu68 (5.3 angstrom) near the protonated Schiff base. Here, we investigate mutant GtACR1s (e.g., E68Q/D234N) expressed in HEK293 cells. The influence of the acidic residues on the absorption wavelengths was also analyzed using a quantum mechanical/molecular mechanical approach. The calculated protonation pattern indicates that Asp234 is deprotonated and Glu68 is protonated in the original crystal structures. The D234E mutation and the E68Q/D234N mutation shorten and lengthen the measured and calculated absorption wavelengths, respectively, which suggests that Asp234 is deprotonated in the wild-type GtACR1. Molecular dynamics simulations show that upon mutation of deprotonated Asp234 to asparagine, deprotonated Glu68 reorients toward the Schiff base and the calculated absorption wavelength remains unchanged. The formation of the proton transfer pathway via Asp234 toward Glu68 and the disconnection of the anion conducting channel are likely a basis of the gating mechanism

Platelet Serotonin (5-HT)2A Receptor Binding Sites in Affective Disorders: A Quantitative Receptor Autoradiographic Study with [ 125I ] Lysergic Acid Diethylamide

We used the quantitative receptor autoradiographic method with a radioligand of [ 125I ]lysergic acid diethylamide ([125I]LSD) to quantitate platelet serotonin (5-HT)2A receptors in affective disorders. Specific binding of [125I]LSD to human platelet pellet sections was saturable, and of high affinity and single. Both ketanserin and spiperone, 5-HT2A selective ligands, inhibited [125I]LSD binding to human platelet pellets with high potency (IC50 values of 0.15 and 0.19 nM, respectively), whereas 5-HT and paroxetine, selective 5- HT re-uptake inhibitors, inhibited binding with a very low potency. These data confirmed that binding sites of human platelet pellets specifically labelled by [125I] LSD were 5-HT2A receptors. The number of 5-HT2A receptors (Bmax of [ 125I] LSD binding) of human platelets obtained from drug-free depressed patients was significantly higher than those of healthy volunteers. There were no statistical differences in the number of 5-HT2A receptors between depressed patients with and without suicidal behaviors. The increased number in platelet 5-HT2A receptotrs may indicate a hyperfunction of the central 5-HT2A receptors. The method with human platelets pellet sections we used is simple and sensitive for investigating platelet 5-HT2A receptors, a diagnostic and therapeutic marker in depressive disorders, in the clinical research

Relaxin-3-Deficient Mice Showed Slight Alteration in Anxiety-Related Behavior

Relaxin-3 is a neuropeptide belonging to the relaxin/insulin superfamily. Studies using rodents have revealed that relaxin-3 is predominantly expressed in neurons in the nucleus incertus (NI) of the pons, the axons of which project to forebrain regions including the hypothalamus. There is evidence that relaxin-3 is involved in several functions, including food intake and stress responses. In the present study, we generated relaxin-3 gene knockout (KO) mice and examined them using a range of behavioral tests of sensory/motor functions and emotion-related behaviors. The results revealed that relaxin-3 KO mice exhibited normal growth and appearance, and were generally indistinguishable from wild genotype littermates. There was no difference in bodyweight among genotypes until at least 28 weeks after birth. In addition, there were no significant differences between wild-type and KO mice in locomotor activity, social interaction, hot plate test performance, fear conditioning, depression-like behavior, and Y-maze test performance. However, in the elevated plus maze test, KO mice exhibited a robust increase in the tendency to enter open arms, although they exhibited normal performance in a light/dark transition test and showed no difference from wild-type mice in the time spent in central area in the open field test. On the other hand, a significant increase in the acoustic startle response was observed in KO mice. These results indicate that relaxin-3 is slightly involved in the anxiety-related behavior

Development of the calibration method for a fast steering antenna for investigating the mode conversion window used in EBW heating in the LHD plasma

In this study, we developed a calibration method for a fast steering antenna for investigating the mode conversion window used in electron Bernstein wave heating in the large helical device. The calibration was carried out in under-dense plasma against a line-of-sight with an optical thickness which varied spatially. Although multi-reflected background radiation becomes dominant in optically thin lines-of-sight, we succeeded in calibrating the fast steering antenna by including the effect of multi-reflected background radiation in the solution of the radiation transfer equation as the constant by which the temperature of the center of the plasma is multiplied. In addition, we report the initial results of experiments investigating the mode conversion window in over-dense plasma using the calibrated antenna

Ethanol Preference and Drinking Behavior Are Controlled by RNA Editing in the Nucleus Accumbens

RNA editing plays critical roles in normal brain function, and alteration of its activity causes various disorders. We previously found that chronic consumption of ethanol was associated with increased levels of RNA editing of serotonin 2C receptor in the nucleus accumbens (NAc). However, it remains unknown whether RNA editing in the NAc modulates alcohol addiction through the brain reward system. To investigate the involvement of NAc RNA editing in alcohol addiction, we generated NAc-specific knockout mice of the double-stranded RNA-specific adenosine deaminase ADAR2 using AAV-GFP/Cre and conducted a battery of behavioral tests including anxiety- and depression-like behaviors. In addition, NAc-specific ADAR2 knockout mice were exposed to ethanol vapor for 20 days, followed by ethanol-drinking and conditioned place preference (CPP) tests. NAc-specific ADAR2 knockout mice showed a significant decrease in locomotor activity in the open field test although they did not develop anxiety- and depression-like behaviors. In addition, the enhancements of ethanol intake and ethanol preference that are usually observed after chronic ethanol vapor exposure were significantly reduced in these mice. These results suggest that ADAR2-mediated RNA editing in the NAc is involved in determination of alcohol preference after chronic alcohol consumption

Safety of Postoperative Administration of Human Urinary Trypsin Inhibitor in Lung Cancer Patients with Idiopathic Pulmonary Fibrosis

Patients with idiopathic pulmonary fibrosis (IPF) undergoing pulmonary resection for lung cancer carry risks of acute exacerbations of IPF (AE) postoperatively. Currently, agents which may attenuate AE are actively sought. Urinary trypsin inhibitor, ulinastatin, is a synthetic glycoprotein which may potentially inhibit various inflammatory factors associated with the development and progression of IPF. The present study was done to evaluate the effects of administration of high dose ulinastatin in lung cancer patients with IPF immediately following lung resection.Patients with IPFs radiologically diagnosed on high resolution CT, and histologically diagnosed resectable lung cancers, were eligible for the study. The effects of escalating doses of ulinastatin 3×10(5), 6×10(5), and 9×10(5) units/body/day, administered postoperatively for 3 days were evaluated. The endpoints were safety and feasibility.Nine patients were evaluated, in cohorts of 3 patients per dosage. Postoperative follow up ranged from 3 to 12 months (median 9 months). The postoperative courses were uneventful in all patients. No subjective adverse events such as abdominal symptoms or skin rashes, or objective adverse events as per serum laboratory tests, such as liver or kidney dysfunctions potentially attributable to ulinastatin administration were observed. AE was seen in one patient at 3 months after surgery, but since this occurred shortly after administration of chemotherapy, it was considered to be attributable to the chemotherapy rather than surgery.Ulinastatin administration after lung resection in lung cancer patients with IPF was considered to be safe and feasible. Further study is planned at the highest dose of this study to evaluate efficacy.UMIN.ac.jp/ctr/UMIN000002410

Improved performance of electron cyclotron resonance heating by perpendicular injection in the Large Helical Device

A real-time interlock system for power injection in electron cyclotron resonance heating(ECRH) was developed to be applied to Large Helical Device (LHD) plasma. This systemenabled perpendicular injection, thus improving the performance of ECRH more than has everbeen achieved before in LHD. Perpendicular propagation of the electron cyclotron wave at77 GHz became more insensitive to the effect of refraction in comparison to the conventionaloblique propagation. The achieved central electron temperature in the case of perpendicularinjection was approximately 2 keV higher than that in the case of standard oblique injectionfor a central electron density of 1 × 1019 m−3 by 1 MW injection.With such improvedperformance of ECRH, high-density ECRH plasma of 8 × 1019 m−3 was successfullysustained after the injection of multiple hydrogen ice pellets for the first time in LHD

Extended capability of the integrated transport analysis suite, TASK3D-a, for LHD experiment

The integrated transport analysis suite, TASK3D-a (Analysis), has been developed to be capable for routine whole-discharge analyses of plasmas confined in three-dimensional (3D) magnetic configurations such as the LHD. The routine dynamic energy balance analysis for NBI-heated plasmas was made possible in the first version released in September 2012. The suite has been further extended through implementing additional modules for neoclassical transport and ECH deposition for 3D configurations. A module has also been added for creating systematic data for the International Stellarator–Heliotron Confinement and Profile Database. Improvement of neutral beam injection modules for multiple-ion species plasmas and loose coupling with a large-simulation code are also highlights of recent developments

Development and application of a ray-tracing code integrating with 3D equilibrium mapping in LHD ECH experiments

The central electron temperature has successfully reached up to 7.5 keV in large helical device(LHD) plasmas with a central high-ion temperature of 5 keV and a central electron density of1.3×1019 m−3. This result was obtained by heating with a newly-installed 154 GHz gyrotronand also the optimisation of injection geometry in electron cyclotron heating (ECH). Theoptimisation was carried out by using the ray-tracing code ‘LHDGauss’, which was upgradedto include the rapid post-processing three-dimensional (3D) equilibrium mapping obtainedfrom experiments. For ray-tracing calculations, LHDGauss can automatically read the relevantdata registered in the LHD database after a discharge, such as ECH injection settings (e.g.Gaussian beam parameters, target positions, polarisation and ECH power) and Thomsonscattering diagnostic data along with the 3D equilibrium mapping data. The equilibrium mapof the electron density and temperature profiles are then extrapolated into the region outsidethe last closed flux surface. Mode purity, or the ratio between the ordinary mode and theextraordinary mode, is obtained by calculating the 1D full-wave equation along the directionof the rays from the antenna to the absorption target point. Using the virtual magnetic fluxsurfaces, the effects of the modelled density profiles and the magnetic shear at the peripheralregion with a given polarisation are taken into account. Power deposition profiles calculatedfor each Thomson scattering measurement timing are registered in the LHD database. Theadjustment of the injection settings for the desired deposition profile from the feedbackprovided on a shot-by-shot basis resulted in an effective experimental procedure

Isotope effects on energy, particle transport and turbulence in electron cyclotron resonant heating plasma of the Large Helical Device

Positive isotope effects have been found in electron cyclotron resonant heating plasma of the Large Helical Device (LHD). The global energy confinement time (τE) in deuterium (D) plasma is 16% better than in hydrogen (H) plasma for the same line-averaged density and absorption power. The power balance analyses showed a clear reduction in ion energy transport, while electron energy transport does not change dramatically. The global particle confinement time (τp) is degraded in D plasma; τp in D plasma is 20% worse than in H plasma for the same line-averaged density and absorption power. The difference in the density profile was not due to the neutral or impurity sources, but rather was due to the difference in the transport. Ion scale turbulence levels show isotope effects. The core turbulence (ρ  =  0.5–0.8) level is higher in D plasma than in H plasma in the low collisionality regime and is lower in D plasma than in H plasma. The density gradient and collisionality play a role in the core turbulence level