Effects of Washcoat on Initial PM Filtration Efficiency and Pressure Drop in SiC DPF

The washcoat (W/C) on a catalytic diesel particulate filter (DPF) greatly alters the pore structure and wall surface condition of the original substrate of DPF, which then affects the filtration efficiency and pressure drop behavior. In the present study, we examined this W/C effect on the initial PM filtration efficiency and pressure loss by changing amounts of washcoat on a SiC-DPF. We measured particle number concentration and particle size distribution in the diesel exhaust gas downstream of the DPF by EEPS. High filtration efficiency was achieved quickly when the W/C amount was increased. We introduced new parameters, T90 and T99, which were the filtration efficiencies that reach more than 90% and 99%, respectively, of the initial DPF usage. The PM trapping mechanisms could be classified according to the PM size. Trapping of PM whose diameter is smaller than 30 nm is little affected by the W/C. However, for PM over 30 nm, as amounts of W/C are increased, particle number concentration decreases rapidly and less PM leakage is observed for the initial filtration process. On the other hand, the initial backpressure and the backpressure during soot loading increased in accordance with W/C amount. Since the relationship between the filtration efficiency and the pressure loss has a trade-off, it is important to design DPF by considering W/C effects.Technical Papers presented at SAE 2011 World Congress & Exhibitionresearch repor

固定資産の管理に関する経営的考察

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THERMAL DEPENDENCE OF SPIN-ORBIT TORQUE-INDUCED MAGNETIZATION REVERSAL IN PERMALLOY THIN FILM

The magnetization of a nanosized thin film reverses owing to the spin-orbit torque (SOT) induced via a spin injection from a heavy metal (HM). The magnetization reversal can be used as a magnetic-memory binary digit at temperatures above room temperature. We investigated magnetization reversal by means of SOT under the thermal effect using micromagnetic simulation. The thermal effect was introduced as random magnetic fields. The averaged magnetization reversal time decreased with increasing temperature. However, several samples exhibited exceptional oscillatory behavior of the magnetization during the reversal and the reversal times increased.This work was supported by the Japan Society for the Promotion of Science KAKENHI Grant Number JP20H02607 and the Kansai University Fund for Supporting Outlay Research Centers 2020.(研究課題「高速磁化反転技術の開発と省エネルギー動作デバイス応用」)departmental bulletin pape

Extraskeletal Mesenchymal Chondrosarcoma : An Immunohistochemical and Ultrastructural Study

This report describes an extremely rare case of extraskeletal mesenchymal chondrosarcoma. The tumor, occurring in the right leg of a male aged 35, was composed of undifferentiated mesenchymal cells and interspersed islands of well-differentiated cartilaginous tissue. Immunohistochemically, S-100 protein was detected in transitional forms between undifferentiated mesenchymal cells and well-differentiated cartilaginous cells as well as well-differentiated cartilaginous cells. Ultrastructurally, the undifferentiated mesenchymal cells had a narrow cytoplasm with a sparsity of organelles. The well-differentiated cartilaginous cells showed many features common to chondrocytes, such as abundant rough endoplasmic reticulum, multiple well-developed Golgi complexes, and microvillous and scalloped cytoplasmic membranes. The differentiation toward cartilaginous cells of undifferentiated mesenchymal cells was indicated by immunohistochemistry and electron microscopy.departmental bulletin pape

世界協学。:留学生50％。外国人教員50％。世界50ヵ国から集まった学生たちとともに学ぶ。

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Aberrant autophagosome and autolysosome formation in <i>spns1</i>-mutant zebrafish.

<p>(<b>A</b>) Yolk opaqueness and LC3 puncta formation in <i>spns1</i>-mutant zebrafish embryos. For EGFP-LC3 transgenic <i>spns1</i>-mutant [<i>Tg(CMV:EGFP-LC3);spns1^hi891/hi891</i>] fish siblings, bright-field and fluorescence images of wild-type (<i>wt</i>) control (upper) and <i>spns1</i> mutant (<i>spns1^−/−</i>) (lower) embryos at 84 hpf are shown. The black arrow indicates the yolk-opaqueness phenotype in the <i>spns1</i> mutant. The gross expression of EGFP-LC3 at head and trunk in the <i>spns1</i>-mutant animal is relatively stronger than in the <i>wt</i> animal. Occasionally, however, a high intensity signal can be observed at the liver region in the mutant (as seen in <b>D</b>). Scale bar, 250 µm. (<b>B</b>) EGFP-LC3 punctate compartments in the liver cells of the <i>spns1</i> mutant. Through high magnification (×600) confocal microscopy, intracellular EGFP-LC3 puncta were visualized in live animals at 84 hpf. Nuclei were counterstained with Hoechest 33342 (blue), and peri-nuclear EGFP-LC3 puncta were evident in the <i>spns1</i> mutant, but not in <i>wt</i> animals. Scale bar, 10 µm. (<b>C</b>) Immunoblotting to detect the conversion of LC3-I to LC-II. Using an anti-LC3 antibody, both endogenous LC3 and transgenic (exogenous) EGFP-LC3 expression was detected and an increase of LC3-II conversion/accumulation was seen in the <i>spns1</i> mutant compared with <i>wt</i> fish at 84 hpf. (<b>D</b>–<b>F</b>) Identification of autophagosome and autolysosome/lysosome formation in the <i>spns1</i> mutant. (D, E) LysoTracker (DND-99; red) staining of EGFP-LC3 transgenic <i>spns1</i>-mutant [<i>Tg(CMV:EGFP-LC3); spns1^hi891/hi891</i>] embryos was performed at 84 hpf. At the whole animal levels (D), the EGFP-LC3 signal is relatively higher throughout in the <i>spns1</i> mutant than in wild type, and a particularly strong signal can be seen in the liver, as shown in (A). In the head and trunk portions of the animals (D), a distinctive increase in the intensity of LysoTracker can be observed in the <i>spns1</i> mutant. At the intracellular level (E), several small LC3 spots and largely diffuse green signal in the cells and cytosolic LysoTracker staining is seen. A number of enlarged LC3- and LysoTracker-positive yellow punctate structures can be seen in the <i>spns1</i> mutant by confocal microscopy at a higher magnification (inset; enlarged from dotted square area). (F) EGFP-LC3 and mCherry-LC3 double-transgenic [<i>Tg(EGFP-LC3:mCherry-LC3)</i>] zebrafish were used to monitor autolysosome formation in <i>spns1</i> MO-injected embryos at 84 hpf. A number of enlarged yellow LC3 puncta were detected in the <i>spns1</i> morphant, while only small yellow LC3 spots can be seen in control-injected embryos. Nuclei were counterstained with 4′, 6-diamidino-2-phenylindole, dihydrochloride (DAPI). Scale bar, 250 µm in (D). Scale bar, 10 µm in (E, F). Quantification of data presented in D (n = 12), E (n = 6), and F (n = 6) is shown in the right graph; the number (n) of animals is for each genotype. Three independent areas (periderm or basal epidermal cells above the eye) were selected from individual animals. (<b>G</b>) Transgenic expression of mCherry-Lamp1 in <i>wt</i> [<i>Tg(CMV:EGFP-LC3)</i>] and <i>spns1</i>-mutant [<i>Tg(CMV:EGFP-LC3);spns1^hi891/hi891</i>] animals 84 hpf. Scale bar, 10 µm. (<b>H</b>) Transgenic expression of EGFP-Vector (<i>vector</i>), EGFP-wild-type Spns1 (<i>spns1 WT</i>), or EGFP-mutant Spns1 (<i>spns1 E153K</i>) in [<i>Tg(CMV:mCherry-LC3);spns1^hi891/hi891</i>] animals at 84 hpf. Scale bar, 10 µm. Quantification of data presented in H is shown for ratio of yolk opaqueness phenotype (n = 48), mCherry intensity (red) (n = 6), and merge intensity of EGFP and mCherry (yellow) (n = 6) in the right graphs; the number (n) of animals is for each genotype. Three independent areas (periderm or basal epidermal cells above the eye) were selected from individual animals. Error bars represent the mean ± standard deviation (S.D.), *<i>p</i><0.005; ns, not significant.</p

Schematic model for Spns1 function under the control of the network module of autophagy-senescence signaling cascades differentially regulated through Beclin 1 and p53.

<p>(<b>A</b>) Beclin 1 is essential for the early stage of autophagy and its depletion suppresses the Spns1 defect by blocking the ‘autophagic process’ and its progression. BafA can decelerate ‘lysosomal biogenesis’, which subsequently presumably prevents autophagosome-lysosome fusion, through the inhibition of the v-ATPase, and contributes to amelioration of the Spns1 defect at least temporarily. Basal p53 activity may suppress the intersection between the ‘autophagic progress’ and ‘lysosomal biogenesis’ where the Beclin 1 depletion was not sufficient, but the v-ATPase inhibition was still effective enough, to compete with the p53 loss to suppress the Spns1 deficiency. By switching the basal p53 state to the activated version with UV irradiation, p53 can promote autophagy. Spns1 might be a gatekeeper of autolysosomal maturation followed by lysosomal biogenesis. It remains unknown how p53 can mechanistically be linked to the lysosomal ‘efflux’ function of Spns1 as well as the lysosomal ‘influx’ function of v-ATPase, and further investigations will be required to explore this connection. (<b>B</b>) Roles of Spns1, p53 and Beclin 1 in senescence equilibrium. Loss of Spns1 leads to an imbalance in homeostasis and increased senescence. This effect can be ameliorated by concurrent knockdown of Beclin 1. p53 has a comparatively less dramatic impact on Spns1-loss-induced embryonic senescence. When in the “basal” state, p53 helps retain equilibrium. When p53 is “activated” by UV irradiation, a modest increase in senescence is observed. The higher level of senescence is seen during loss of Spns1 in the absence of basal p53 or in the presence of activated p53. During loss/knockdown of all three genes (<i>spns1</i>, <i>p53</i> and <i>beclin 1</i>), a state of moderate senescence is observed. An increase in senescence is accompanied by a p53-dependent decrease in cellular proliferation.</p

Knockdown of <i>beclin 1</i> suppresses abnormal autolysosomal puncta formation and embryonic senescence caused by Spns1 deficiency in zebrafish.

<p>(<b>A</b>) Effect of <i>beclin 1</i> knockdown on EGFP-LC3 puncta formation in <i>spns1</i>-depleted zebrafish embryos. Injection of control (water) injection, <i>spns1</i> MO (4 ng/embryo) or coinjection of <i>spns1</i> MO (4 ng/embryo) and <i>beclin 1</i> MO (12 ng/embryo) into <i>Tg(CMV:EGFP-LC3)</i> fish was performed to assess whether the <i>beclin 1</i> knockdown reduces or eliminates aggregated LC3 puncta induced by Spns1 depletion at 84 hpf. Scale bar, 10 µm. Quantification of data presented in panel A (n = 12) is shown in the right graph; the number (n) of animals is for each morphant or water-injected control. Three independent areas (periderm or basal epidermal cells above the eye) were selected from individual animals. (<b>B</b>) Effect of <i>beclin 1</i> knockdown on EGFP-GABARAP as well as mCherry-LC3 puncta formation in <i>spns1</i>-depleted zebrafish embryos. Injection of control (water), <i>spns1</i> MO or coinjection of <i>spns1</i> MO and <i>beclin 1</i> MO into <i>Tg(CMV:EGFP-GABARAP;mCherry-LC3)</i> fish was performed to evaluate whether the <i>beclin 1</i> knockdown reduces or eliminates the aggregation of GFP-GABARAP puncta in comparison with those of LC3 caused by the Spns1 depletion at 84 hpf. Scale bar, 10 µm. Quantification of data presented in the top row (green; EGFP) (n = 9), middle row (red; mCherry) (n = 12), and bottom row (yellow; merge of EGFP and mCherry) (n = 9) in panel B is shown in the right graphs; the number (n) of animals is for each morphant or water-injected control. Three independent areas (periderm or basal epidermal cells above the eye) were selected from individual animals. (<b>C</b>) Effect of <i>beclin 1</i> knockdown on embryonic senescence in <i>spns1</i> morphant. By using the same injection samples [injection of control (water), <i>spns1</i> MO or coinjection of <i>spns1</i> MO and <i>beclin 1</i> MO into <i>Tg(CMV:EGFP-GABARAP;mCherry-LC3)</i> fish], SA-β-gal staining was performed to assess whether the <i>beclin 1</i> knockdown has any impact on the embryonic senescence caused by Spns1 depletion at 84 hpf. Representative images of individual fish by bright field (BF, live samples) and SA-β-gal (SABG) staining are shown in the upper and middle panels, respectively. Scale bar, 250 µm. Lower panels are larger magnification images of corresponding SA-β-gal samples shown in the middle panels and the fluorescence images of nuclei counterstained with DAPI. Scale bar, 10 µm. Quantification of data presented in the middle row (SABG) in panel C (n = 12) is shown in the right graph; the number (n) of animals is for each morphant or water-injected control. Error bars represent the mean ± S.D., *<i>p</i><0.005.</p

Acidity-dependent lysosomal biogenesis is rate limiting in <i>spns1</i>-mutant zebrafish.

<p>(<b>A</b>) Effect of bafilomycin A₁ (BafA) on the yolk opaque phenotype (BF; bright field) and embryonic senescence (SABG; SA-β-gal) in the <i>spns1</i> mutant in the presence or absence of <i>p53</i> at 48 hpf. Normal wild-type (<i>spns1^+/+;p53^+/+</i>), <i>tp53^zdf1/zdf1</i> (<i>p53^m/m</i>), <i>spns1^hi891/hi891</i> (<i>spns1^−/−</i>) and <i>spns1^hi891/hi891;tp53^zdf1/zdf1</i> (<i>spns1^−/−;p53^m/m</i>) embryos at 36 hpf were incubated with BafA (200 nM) for 12 h, and stained with LysoTracker at 48 hpf, followed by SA-β-gal staining at 60 hpf. Scale bar, 250 µm. (<b>B</b>) Quantification of the SA-β-gal intensities shown in (A). Quantification of data presented in panel A (n = 12) is shown; the number (n) of animals is for each genotype with DMSO or BafA. (<b>C</b>) Gross morphology, EGFP-LC3 and LysoTracker intensities in wild-type (<i>wt</i>) and <i>spns1</i>-mutant animals treated with BafA shown at 48 hpf (12 h treatment starting at 36 hpf). Scale bar, 250 µm. (<b>D</b>) Quantification of the EGFP-LC3 and LysoTracker fluorescence intensities shown in (C). Quantification of data presented in the middle and bottom rows (green; EGFP, red; mCherry) in panel C (n = 12) is shown; the number (n) of animals is for each genotype with DMSO or BafA. (<b>E</b>) Intracellular autolysosome formation and lysosomal biogenesis in the BafA-treated <i>spns1</i> mutant. The samples analyzed in (C) were observed by using confocal microscopy at high magnification (×600). Scale bar, 10 µm. (<b>F</b>) Quantification of the EGFP-LC3 and LysoTracker fluorescence intensities shown in (E). Quantification of data presented for EGFP (green) and mCherry (red) signals in panel E (n = 6) is shown; the number (n) of animals is for each genotype with DMSO or BafA. Three independent areas (periderm or basal epidermal cells above the eye) were selected from individual animals. (<b>G</b>) Insufficient intracellular acidity constituent in the <i>spns1</i> mutants. Using two different acidic-sensitive probes, LysoSensor 189 and neutral-sensitive LysoSensor 153 (green), in combination with LysoTracker (red), <i>wt</i> and <i>spns1</i>-mutant animals showed detectable signals when stained at 72 hpf. In <i>spns1</i>-mutant animals, autolysosomal and/or lysosomal compartments were more prominently detectable by LysoSensor 153 than by LysoSensor 189, at the cellular level with enhanced signal intensity of these enlarged compartments. In stark contrast, the cellular compartments in <i>wt</i> fish treated with pepstatin A and E-64-d (P/E) (12 h treatment from 60 hpf through 72 hpf) were more prominently detectable by LysoSensor 189 than by LysoSensor 153 under the identical LysoTracker staining conditions. Of note, these autolysosomal and lysosomal compartments in <i>spns1</i> mutants, as well as in <i>wt</i> animals treated with pepstatin A and E-64-d, may still retain some weak (higher pH) and strong (lower pH) acidity, respectively, as short-term BafA treatment (for 1 h between 71 and 72 hpf) can abolish the acidic compartments stained by both LysoSensor and LysoTracker (<b><a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1004409#pgen.1004409.s017" target="_blank">Figure S17C and D</a></b>). Scale bar, 10 µm. (<b>H</b>) Quantification of the LysoSensor (189 and 153) and LysoTracker fluorescence intensities shown in (G). Quantification of data presented for LysoSensor (green) and LysoTracker (red) signals in panel G (n = 6) is shown; the number (n) of animals is for each genotype with DMSO or pepstatin A and E-64-d (P/E). Three independent areas (periderm or basal epidermal cells above the eye) were selected from individual animals. Error bars represent the mean ± S.D., *<i>p</i><0.005; ns, not significant.</p

p53 depletion does not suppress but rather exacerbates Spns1 deficiency.

<p>(<b>A</b>) Effect of <i>p53</i> knockdown on embryonic senescence and autolysosome formation in <i>spns1</i> morphants. The impact of transient <i>p53</i> knockdown on SA-β-gal (SABG) induction, as well as on EGFP-LC3 and LysoTracker (LysoT) puncta, was determined in <i>spns1</i> morphants at 84 hpf, followed by the MO (4 ng/embryo) injections. Inverse-sequence <i>p53</i> MO (inv. <i>p53</i> MO) was used as a negative control for the original <i>p53</i> MO. Scale bar, 250 µm in the SABG images. Scale bar, 10 µm in the fluorescence images. (<b>B</b>) Quantification of the SA-β-gal intensities in MO-injected animals, as shown for the SABG images in (A). Quantification of data presented in the top row (SABG) in B (n = 12) is shown; the number (n) of animals is for each morphant. (<b>C</b>) Quantification of EGFP-LC3 and LysoTracker puncta in MO-injected animals shown in (A) (n = 9); the number (n) of animals is for each morphant. Three independent areas (periderm or basal epidermal cells above the eye) were selected from individual animals. (<b>D</b>) Effect of a <i>p53</i> mutation on embryonic SA-β-gal activity in the <i>spns1</i> mutant. The heritable impact of p53 and Spns1 on SA-β-gal induction was tested in each single gene mutant [<i>spns1^hi891/hi891</i> (<i>spns1^−/−</i>) or <i>tp53^zdf1/zdf1</i> (<i>p53^m/m</i>)] and double mutant <i>spns1^hi891/hi891;tp53^zdf1/zdf1</i> (<i>spns1^−/−;p53^m/m</i>) compared with wild-type (<i>wt</i>) animals at 84 hpf. Scale bar, 250 µm. (<b>E</b>) Quantification of the SA-β-gal intensities in <i>wt</i>, <i>tp53^zdf1/zdf1</i>, <i>spns1^hi891/hi891</i> and <i>spns1^hi891/hi891;tp53^zdf1/zdf1</i> animals, shown in (D). Quantification of data presented in panel D (n = 12) is shown; the number (n) of animals is for each genotype. (<b>F</b>) Quantitative RT-PCR analyses of senescence marker and/or mediator expression as well as p53-downstream target genes in <i>wt</i>, <i>tp53^zdf1/zdf1</i>, <i>spns1^hi891/hi891</i> and <i>spns1^hi891/hi891;tp53^zdf1/zdf1</i> at 72 hpf. Data are mean ±SD [n = 4 samples (3 embryos/sample) per genotype]. Asterisks denote significant changes compared to <i>wt</i> values. *<i>p</i><0.05. (<b>G</b>) LC3 conversions in <i>p53</i> and <i>spns1</i>-mutant animals. Protein detection for the conversion/accumulation of LC3-I to LC-II was performed in the described mutant background animals in comparison with <i>wt</i> fish at 84 hpf. Western blot analysis using anti-LC3 antibody shows endogenous LC3 protein levels, which can confirm an increase of the total amount of LC3 in the <i>p53</i> mutant compared with <i>wt</i> fish. Increased LC3-II conversion/accumulation was detected in <i>p53</i> and <i>spns1</i> double-mutants as well as in <i>spns1</i> single-mutant fish. (<b>H</b>) The blotting band intensities of LC3-I, LC3-II and β-actin were quantified (n = 6), and the relative ratios between LC3-II/actin and LC3-I/actin are shown in the bar graph; the number (n) of animals is for each genotype. (<b>I</b>) <i>wt</i>, <i>tp53^zdf1/zdf1</i>, <i>spns1^hi891/hi891</i> and <i>spns1^hi891/hi891;tp53^zdf1/zdf1</i> embryos injected with <i>beclin 1</i> MO or control MO (12 ng/embryo) were assayed for SA-β-gal at 84 hpf. <i>beclin 1</i> MO-mediated suppression of SA-β-gal in <i>spns1^hi891/hi891</i> animals was attenuated in the <i>p53</i> mutant background. Scale bar, 250 µm. (<b>J</b>) Quantification of the SA-β-gal intensities shown in (I). Quantification of data presented in H (n = 12) is shown; the number (n) of animals is for each genotype with MO. Error bars represent the mean ± S.D., *<i>p</i><0.005; ns, not significant.</p