Utility of syntenic relationships of VDAC1 pseudogenes for not only an understanding of the phylogenetic divergence history of rodents, but also ascertaining possible pseudogene candidates as genuine pseudogenes

Rodent and human genomes were screened to identify pseudogenes of the type 1 voltage-dependent anion channel (VDAC1) in mitochondria. In addition to the 16 pseudogenes of rat VDAC1 identified in our recent study, 15 and 13 sequences were identified as pseudogenes of VDAC1 in mouse and human genome, respectively; and 4, 2, and 1 sequences, showing lower similarities with the VDAC1 sequence, were identified as “possible pseudogene candidates” in rat, mouse, and human, respectively. No syntenic combination was observed between rodent and human pseudogenes, but 2 and 1 possible pseudogene candidates of VDAC1 of rat and mouse, respectively, were found to have syntenic counterparts in mouse and rat genome, respectively; and these syntenic counterparts were genuine VDAC1 pseudogenes. Therefore, syntenic combinations of pseudogenes of VDAC1 were useful not only for a better understanding of the phylogenetic divergence history of rodents but also for ascertaining possible pseudogene candidates as genuine pseudogenes

Glutathione adducts induced by ischemia and deletion of glutaredoxin-1 stabilize HIF-1α and improve limb revascularization

Reactive oxygen species (ROS) are increased in ischemic tissues and necessary for revascularization; however, the mechanism remains unclear. Exposure of cysteine residues to ROS in the presence of glutathione (GSH) generates GSH-protein adducts that are specifically reversed by the cytosolic thioltransferase, glutaredoxin-1 (Glrx). Here, we show that a key angiogenic transcriptional factor hypoxia-inducible factor (HIF)-1α is stabilized by GSH adducts, and the genetic deletion of Glrx improves ischemic revascularization. In mouse muscle C2C12 cells, HIF-1α protein levels are increased by increasing GSH adducts with cell-permeable oxidized GSH (GSSG-ethyl ester) or 2-acetylamino-3-[4-(2-acetylamino-2-carboxyethylsulfanyl thiocarbonylamino) phenylthiocarbamoylsulfanyl] propionic acid (2-AAPA), an inhibitor of glutathione reductase. A biotin switch assay shows that GSSG-ester-induced HIF-1α contains reversibly modified thiols, and MS confirms GSH adducts on Cys520 (mouse Cys533). In addition, an HIF-1α Cys520 serine mutant is resistant to 2-AAPA–induced HIF-1α stabilization. Furthermore, Glrx overexpression prevents HIF-1α stabilization, whereas Glrx ablation by siRNA increases HIF-1α protein and expression of downstream angiogenic genes. Blood flow recovery after femoral artery ligation is significantly improved in Glrx KO mice, associated with increased levels of GSH-protein adducts, capillary density, vascular endothelial growth factor (VEGF)-A, and HIF-1α in the ischemic muscles. Therefore, Glrx ablation stabilizes HIF-1α by increasing GSH adducts on Cys520 promoting in vivo HIF-1α stabilization, VEGF-A production, and revascularization in the ischemic muscle

Measurement of electrostatic potential fluctuation using heavy ion beam probe in large helical device

Heavy ion beam probe (HIBP) for large helical device (LHD) has been improved to measure the potential fluctuation in high-temperature plasmas. The spatial resolution is improved to about 10 mm by controlling the focus of a probe beam. The HIBP is applied to measure the potential fluctuation in plasmas where the rotational transform is controlled by electron cyclotron current drive. The fluctuations whose frequencies change with the time constant of a few hundreds of milliseconds and that with a constant frequency are observed. The characteristics of the latter fluctuation are similar to those of the geodesic acoustic mode oscillation. The spatial profiles of the fluctuations are also obtained

Present Status in the Development of 6 MeV Heavy Ion Beam Probe on LHD

In order to measure the potential in Large Helical Device (LHD), we have been developing a heavy ion beam probe (HIBP). For probing beam, gold beam is used, which is accelerated by a tandem accelerator up to the energy of 6 MeV. The experiments for calibration of beam orbit were done, and experimental results were compared with orbit calculations. The experimental results coincided fairly with the calculation results. After the calibration of the beam orbit, the potential in plasma was tried to measure with the HIBP. The experimental data showed positive potential in a neutral beam heating phase on the condition of ne ? 5 × 10^18 m^-3, and the increase of potential was observed when the additional electron cyclotron heating was applied to this plasma. The time constant for this increase was about a few tens ms, which was larger than a theoretical expectation. In the spatial position of sample volume, we might have an ambiguity in this experiment

Acute Activation of AMP-Activated Protein Kinase Prevents H2O2-Induced Premature Senescence in Primary Human Keratinocytes

We investigated the effects of AMPK on H2O2-induced premature senescence in primary human keratinocytes. Incubation with 50 µM H2O2 for 2 h resulted in premature senescence with characteristic increases in senescence-associated ß-galactosidase (SA-gal) staining 3 days later and no changes in AMPK or p38 MAPK activity. The increase in SA-gal staining was preceded by increases in both p53 phosphorylation (S15) (1 h) and transactivation (6 h) and the abundance of the cyclin inhibitor p21CIP1 (16 h). Incubation with AICAR or resveratrol, both of which activated AMPK, prevented the H2O2-induced increases in both SA-Gal staining and p21 abundance. In addition, AICAR diminished the increase in p53 transactivation. The decreases in SA-Gal expression induced by resveratrol and AICAR were prevented by the pharmacological AMPK inhibitor Compound C, expression of a DN-AMPK or AMPK knock-down with shRNA. Likewise, both knockdown of AMPK and expression of DN-AMPK were sufficient to induce senescence, even in the absence of exogenous H2O2. As reported by others, we found that AMPK activation by itself increased p53 phosphorylation at S15 in embryonic fibroblasts (MEF), whereas under the same conditions it decreased p53 phosphorylation in the keratinocytes, human aortic endothelial cells, and human HT1080 fibrosarcoma cells. In conclusion, the results indicate that H2O2 at low concentrations causes premature senescence in human keratinocytes by activating p53-p21CIP1 signaling and that these effects can be prevented by acute AMPK activation and enhanced by AMPK downregulation. They also suggest that this action of AMPK may be cell or context-specific

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

Recent Results from LHD Experiment with Emphasis on Relation to Theory from Experimentalist’s View

he Large Helical Device (LHD) has been extending an operational regime of net-current free plasmas towardsthe fusion relevant condition with taking advantage of a net current-free heliotron concept and employing a superconducting coil system. Heating capability has exceeded 10 MW and the central ion and electron temperatureshave reached 7 and 10 keV, respectively. The maximum value of β and pulse length have been extended to 3.2% and 150 s, respectively. Many encouraging physical findings have been obtained. Topics from recent experiments, which should be emphasized from the aspect of theoretical approaches, are reviewed. Those are (1) Prominent features in the inward shifted configuration, i.e., mitigation of an ideal interchange mode in the configuration with magnetic hill, and confinement improvement due to suppression of both anomalous and neoclassical transport, (2) Demonstration ofbifurcation of radial electric field and associated formation of an internal transport barrier, and (3) Dynamics of magnetic islands and clarification of the role of separatrix

Extension of operational regime in high-temperature plasmas and effect of ECRH on ion thermal transport in the LHD

A simultaneous high ion temperature (Ti) and high electron temperature (Te) regime was successfully extended due to an optimized heating scenario in the LHD. Such high-temperature plasmas were realized by the simultaneous formation of an electron internal transport barrier (ITB) and an ion ITB by the combination of high power NBI and ECRH. Although the ion thermal confinement was degraded in the plasma core with an increase of Te/Ti by the on-axis ECRH, it was found that the ion thermal confinement was improved at the plasma edge. The normalized ion thermal diffusivity ${{\chi}_{\text{i}}}/T_{\text{i}}^{1.5}$ at the plasma edge was reduced by 70%. The improvement of the ion thermal confinement at the edge led to an increase in Ti in the entire plasma region, even though the core transport was degraded