生体深部精緻観察のための透過型液晶補償光学素子を用いた光音響顕微鏡

佐賀大学博士(工学)学位論文(Thesis)doctoral thesi

Temporal and Spatial Variations of Total Electron Content Enhancements During a Geomagnetic Storm on 27 and 28 September 2017

Temporal and spatial evolutions of total electron content (TEC) and electron density in the ionosphere during a geomagnetic storm that occurred on 27 and 28 September 2017 have been investigated using global TEC data obtained from many Global Navigation Satellite System stations together with the ionosonde, geomagnetic field, Jicamarca incoherent scatter and Super Dual Auroral Radar Network (SuperDARN) radar data. Our analysis results show that a clear enhancement of the ratio of the TEC difference (rTEC) first occurs from noon to afternoon at high latitudes within 1 hr after a sudden increase and expansion of the high‐latitude convection and prompt penetration of the electric field to the equator associated with the southward excursion of the interplanetary magnetic field. Approximately 1–2 hr after the onset of the hmF2 increase in the midlatitude and low‐latitude regions associated with the high‐latitude convection enhancement, the rTEC and foF2 values begin to increase and the enhanced rTEC region expands to low latitudes within 1–2 hr. This signature suggests that the ionospheric plasmas in the F2 region move at a higher altitude due to local electric field drift, where the recombination rate is smaller, and that the electron density increases due to additional production at the lower altitude in the sunlit region. Later, another rTEC enhancement related to the equatorial ionization anomaly appears in the equatorial region approximately 1 hr after the prompt penetration of the electric field to the equator and expands to higher latitudes within 3–4 hr.ファイル公開：2021-01-01journal articl

Temporal and Spatial Variations of Total Electron Content Enhancements During a Geomagnetic Storm on 27 and 28 September 2017

Temporal and spatial evolutions of total electron content (TEC) and electron density in the ionosphere during a geomagnetic storm that occurred on 27 and 28 September 2017 have been investigated using global TEC data obtained from many Global Navigation Satellite System stations together with the ionosonde, geomagnetic field, Jicamarca incoherent scatter and Super Dual Auroral Radar Network (SuperDARN) radar data. Our analysis results show that a clear enhancement of the ratio of the TEC difference (rTEC) first occurs from noon to afternoon at high latitudes within 1 hr after a sudden increase and expansion of the high‐latitude convection and prompt penetration of the electric field to the equator associated with the southward excursion of the interplanetary magnetic field. Approximately 1–2 hr after the onset of the hmF2 increase in the midlatitude and low‐latitude regions associated with the high‐latitude convection enhancement, the rTEC and foF2 values begin to increase and the enhanced rTEC region expands to low latitudes within 1–2 hr. This signature suggests that the ionospheric plasmas in the F2 region move at a higher altitude due to local electric field drift, where the recombination rate is smaller, and that the electron density increases due to additional production at the lower altitude in the sunlit region. Later, another rTEC enhancement related to the equatorial ionization anomaly appears in the equatorial region approximately 1 hr after the prompt penetration of the electric field to the equator and expands to higher latitudes within 3–4 hr.ファイル公開：2021-01-01journal articl

N-terminal HEAT repeats of C. albicans Tor1 are required for specific stress responses and for down-modulating TORC1 signaling activity

Presentation of poster 1890C at TAGC 2020 Online.File includes PDF of the poster (TAGC 2020 virtual poster-1890C_Wanjun Qi.pdf) and a 6min34sec video walkthrough of the poster material (Walk through for poster 1890C.m4a)</div

<i>C</i>. <i>albicans</i> Gdh2 is cytosolic but alkalization is dependent on mitochondrial function.

(A) Wildtype cells (SC5314) from overnight YPD cultures were washed and diluted to either OD600 ≈ 0.1 (top panel) or ≈ 5 (bottom panel) in liquid YNB+CAA with the indicated concentrations of mitochondrial complex III inhibitor antimycin A. Cultures were grown at 37°C under constant aeration for 24 h and 2.5 h, respectively, and photographed. To assess viability after antimycin A treatment, inhibited cells from 24 h old culture (top panel) were harvested, washed, and resuspended in fresh YNB+CAA media and incubated for 24 h at 37°C (middle panel). Images are representative of at least 3 independent experiments. (B) Strains expressing either Gdh2-GFP (CFG273) or Put2-GFP (CFG219) were grown overnight in YPD, harvested, washed, resuspended in SED (with 0.2% glucose) or YNB + CAA and grown for 24 h at 37°C, and stained with 200 nM MTR prior to imaging by differential interference contrast (DIC) and confocal fluorescence microscopy; the scale bar = 5 μm. The inset in the Gdh2-GFP panels (higher magnification) shows that the GFP and MTR signals do not overlap. (C) Fractionation of extracts from cells expressing Gdh2-GFP and Put2-HA (CFG397 or CFG404) or Gdh2-GFP and Put1-RFP (CFG398 or CFG407) (upper left panels). Cells, pre-grown in YPD, were shifted to YNB+CAA (OD600 ≈ 2) and grown for 2 h at 37°C. Cytosolic (C) and mitochondrial (M) fractions from total cell lysate (T) were analyzed by immunoblotting using Put2-HA and Tdh3 (GAPDH) as mitochondrial and cytosolic markers, respectively. Gdh2-GFP colocalizes with Tdh3, whereas Put1-RFP colocalizes with Put2-HA. Blots shown are representative of 3 biological replicates. Confocal microscopy of Gdh2-GFP and Put1-RFP fluorescence confirms that Gdh2-GFP exhibits an extramitochondrial localization (upper right panels); Airyscan-processed images—LSM800 confocal microscope, scale bar = 5 μm. The N-terminal regions of Gdh2, Put1, Put2 and Tdh3 were queried for functioning as a mitochondrial targeting sequence (Mitofates; lower panel). The N-termini of Gdh2 and Tdh3, in contrast to Put1 and Put2, have low probability scores. Predicted cleavage sites for mitochondrial processing peptidase (MPP, red font) and mitochondrial intermediate peptidase (Oct1, blue font).</p

Plasma membrane stress responses did not significantly depend on Tor1 N-terminal HEAT repeats.

A. Dilutions of cells of indicated genotypes were spotted on YPD medium without or with doxycycline (300 Doxy) and plasma membrane stress was induced with 0.005% SDS. (TOR1/TOR1, JKC1713; tor1/TOR1, JKC1345, JKC1346, JKC1347; tetO-TOR1-Del381, JKC1442, JKC1445, JKC1441; tetO-TOR1, JKC1543, JKC1546, JKC1549). B. Cells of indicated genotypes were grown in YPD medium containing Vehicle (Veh, H2O) or 0.05% SDS, without or with 300 ng/ml doxycycline. Upper panel shows the actual growth curves and lower panel shows the corresponding area under the curve (AUC) for each strain. ** is p = 0.0076; * is p = 0.029 (Veh, 0 Doxy) and p = 0.0129 (0.05% SDS, 300 Doxy); ns is p = 0.2, error bars show SD of 2 biological replicates (TOR1/TOR1, JKC1713; tor1/TOR1, JKC1347; tetO-TOR1-Del381, JKC1441; tetO-TOR1, JKC1549). C. Cells of indicated genotypes were pre-grown in YPD medium with 5 ng/ml doxycycline for 3.5 h, then diluted into fresh YPD medium containing either Vehicle (Veh, H2O) or 0.01% SDS and incubated for the indicated times. Total protein extract was probed with antibody to phosphorylated Rps6 (P-S6) and tubulin (Tub) as loading control. Dens: signal intensity ratio of P-S6 to Tub; strains as in panel B. D. Cells were grown as in B but treated with 1 μg/ml Fluconazole (Fluc) or 0.2 μg/ml Amphotericin B (AmB).</p

Supplementary Text.

Tables. Table A. C. albicans strains used in this study. Table B. Plasmids used in this study. Table C. Oligonucleotides used in this study. Table D. Antibodies used in this study. (PDF)</p

Comparison of gene set expression changes during repression of <i>TOR1</i> and <i>TOR1-Del381</i> in FL and Del381 cells exposed to doxycycline.

A. Enrichment map showing the major categories of genes identified by Gene Set Enrichment Analysis (GSEA) in the transcript profiles of tor1/tetO-TOR1-Del381 cells (JKC1441, left side) and tor1/tetO-TOR1 cells (JKC1549, right side) compared to heterozygous tor1/TOR1 cells (-/+, JKC1347) following tetO repression in YPD with doxycycline for 2 h. Nodes represent gene sets (FDR q B. Comparison of gene expression changes (≥2-fold up and down) in tor1/tetO-TOR1-Del381 (JKC1441) and tor1/tetO-TOR1 (JKC1549) cells exposed to doxycycline for 8 h. Six genes induced in tor1/tetO-TOR1-Del381 cells were suppressed in tor1/tetO-TOR1 cells, including UME6, ECE1, HYR1 and HWP1. C. Heatmap showing expression of genes that encode Complex I of the mitochondrial electron transport chain in tor1/tetO-TOR1-Del381 (JKC1441) relative to the heterozygote tor1/TOR1 (column 1) and in tor1/tetO-TOR1 (JKC1549) relative to the heterozygote tor1/TOR1 (column 2). Expression values are Log2 values relative to tor1/TOR1 (JKC1347).</p

Genes showing decreased expression in D381 following 2h exposure to Doxycycline.

(XLSX)</p

Tor1 N-terminal HEAT repeats were required for oxidative stress-induced delay of translation initiation.

A. Cells expressing GFP from tetO (ON) (strain TETG25B) were grown in YPD medium for 15 h, then inoculated into fresh YPD and pre-grown for 3.5 h (Time 0, 0 min). GFP expression was induced with 50 μg/ml doxycycline in SC medium with Vehicle (DMSO) or 15 μM Plumbagin (Plum). Negative control (NC) cells were grown in SC medium without doxycycline (with DMSO) for 120 min. Total protein extracts were probed with antibody to GFP, and tubulin (Tub) as loading control. Dens: signal intensity ratio of GFP to Tub. Representative of 2 biological replicates. B. Cells expressing pMAL2-GFP in backgrounds with distinct TOR1 alleles were pre-grown in YPD medium with 10 ng/ml doxycycline for 15 h (+/+, JKC2616; -/+, JKC2620; Del381, JKC2624; FL, JKC2628). GFP expression was induced by inoculation into YP-Maltose medium with 5 ng/ml doxycycline, containing 13 μM Plumbagin (Plum) or DMSO as vehicle (Veh). (0 h, JKC2616 after pre-growth; N, JKC2616 inoculated into YPD instead of YP-Maltose with 5 ng/ml doxycycline, Plumbagin or DMSO, grown for 9 h). Total protein extracts were probed with antibody to GFP, and tubulin (Tub) as loading control. Dens: signal intensity ratio of GFP to Tub.</p