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Discordance between fractional flow reserve (FFR) and instantaneous wave-free ratio (iFR) occurs in approximately 20 % of cases. However, no studies have reported the discordance in patients with severe aortic stenosis (AS). We aimed to evaluate the diagnostic discordance between FFR and iFR in patients with severe AS.We examined 140 consecutive patients with severe AS (164 intermediate coronary artery stenosis vessels). FFR and iFR were calculated in four quadrants based on threshold FFR and iFR values of ≤0.8 and ≤0.89, respectively (Group 1: iFR0.89, FFR0.80; Group 2: iFR ≤0.89, FFR0.80; Group 3: iFR0.89, FFR ≤0.80; and Group 4: iFR ≤0.89, FFR ≤0.80). Concordant groups were Groups 1 and 4, and discordant groups were Groups 2 and 3. Positive and negative discordant groups were Groups 3 and 2, respectively.The median (Q1, Q3) FFR and iFR were 0.84 (0.76, 0.88) and 0.85 (0.76, 0.91), respectively. Discordance was observed in 48 vessels (29.3 %). In the discordant group, negative discordance (Group 2: iFR ≤0.89 and FFR0.80) was predominant (45 cases, 93.6 %). Multivariate analysis showed that the left anterior descending artery [odds ratio (OR), 3.88; 95 % confidence interval (CI): 1.54-9.79, p = 0.004] and peak velocity ≥5.0 m/s (OR, 3.21; 95%CI: 1.36-7.57, p = 0.008) were independently associated with negative discordance (FFR0.8 and iFR ≤0.89).In patients with severe AS, discordance between FFR and iFR was predominantly negative and observed in 29.3 % of vessels. The left anterior descending artery and peak velocity ≥5.0 m/s were independently associated with negative discordance.

BACKGROUND/PURPOSE Data on the feasibility of coronary access (CA) through above or outside of the cylindrical shaped-transcatheter heart valve (THV) are very limited. The aims of the present study were to assess the feasibility of CA after transcatheter aortic valve replacement (TAVR) with the LOTUS using multi detector computed tomography (MDCT) and the reliability of algorithm detecting unfavorable CA. METHODS/MATERIALS Post-TAVR MDCT of 41 patients with 82 coronary arteries were evaluated. The relationship and distance between the THV flame and sinotubular junction (STJ) and coronary ostia were assessed. Unfavorable CA was defined as the valve-to-STJ distance < 2-mm or the valve-to-coronary ostia distance < 2-mm if the THV flame was above STJ or coronary ostia. RESULTS MDCT-identified unfavorable CA was observed in 29.3% for the left coronary artery and 41.5% for the right coronary artery. In total, 53.7% of patients had at least one unfavorable CA and 14.6% of those had unfavorable CA for both left and right coronary artery. While patients underwent coronary angiography after TAVR, the success rates of selective coronary cannulation were significantly lower in patients with MDCT-identified unfavorable CA in comparison to those with favorable CA for left (20.0% vs. 100%, P = 0.002) and right coronary artery (0% vs. 100%, P < 0.001). CONCLUSIONS Future CA through above or outside of a cylindrical shaped THV may be challenging with a significant probability. Our algorithm identifying unfavorable CA using post-MDCT seems to be useful for estimating the risk of unsuccessful selective cannulation.

This paper provides a possible solution to the issue of whether functional projections are necessary for secondary predicates. Focusing on manner adverbs and resultative predicates in Japanese because they have the same particles -ni or -ku, I will present several pieces of evidence that these two kinds of phrases are of the same syntactic category but should be given different treatments. As a proposal, I argue that, although in Japanese, resultative predicates are syntactic adjuncts, both manner adverbs and resultative predicates are headed by functional projections that take adjectival phrases in their complements and are responsible for the result meaning. I further claim that particles of Japanese resultative predicates are realizations of the functional projection, while adverbial categorizers in the sense of Distributed Morphology are realized as the particles of manner adverbs. It will also be shown that the functional projection is obligatorily realized as -ni or -ku, but this restriction is not imposed on the adverbial categorizer, which undergoes various realizations. The differences between these two phrases can be accounted for by assuming different syntactic structures for these phrases, leading to the conclusion that functional projections are necessary with respect to resultative predicates in Japanese.

Background Difficulties are often encountered while controlling atrial fibrillation (AF), especially in hemodialysis (HD) patients. Previous data revealed that cryoballoon ablation (CBA) for treating paroxysmal atrial fibrillation (PAF) was not inferior to radiofrequency ablation (RFA); however, HD patients were excluded in this prior trial. Thus, the efficacy of CBA for HD patients is still unknown. Methods This retrospective study analyzed HD patients who underwent catheter ablation (CA) for AF from August 2011 to June 2019. Patients who received CBA (CBA group) and those who received RFA (RFA group) were compared. The primary endpoint was defined as freedom from a composite outcome (a documented recurrence of any atrial tachyarrhythmia or a prescription of antiarrhythmic drugs) at one year after CA. Results The RFA and CBA groups were composed of 21 and 23 patients, respectively. Freedom from a composite outcome was 58.4% in the RFA group and 68.2% in the CBA group (Log-rank: p = 0.571). Conclusion Our results suggest that patients on HD with AF who were treated with CBA tended to have better outcomes than patients treated with RFA. Therefore, CBA could be a suitable ablation method for HD patients.

We herein report the cytokine expression at different stages for three patients who developed cardiac complications after immune checkpoint inhibitor (ICI) therapy. Case 1 with biopsy-proven myocarditis showed increased levels of interleukin (IL)-8, monocyte chemotactic and activating factor, and granulocyte macrophage colony-stimulating factor (GM-CSF) when he developed Takotsubo cardiomyopathy. Case 2 with subclinical myocarditis showed predominant activation of IL-8 during the progressive clinical course. Case 3 with cytokine-releasing syndrome showed substantial activations of IL-6, IL-8, GM-CSF, and interferon-γ. Our data suggest the development of unique cytokine activation in individual patients with cardiac complications after ICI therapy.

We have been using sandwich freezing of living yeast and bacteria followed by freeze-substitution for observing close-to-native ultrastructure of cells. Recently, sandwich freezing of glutaraldehyde-fixed cultured cells and human tissues have been found to give excellent preservation of ultrastructure of cells and tissues. These studies, however, have been conducted using a handmade sandwich freezing device and have been limited in a few laboratories. To spread the use of this method to other laboratories, we fabricated and commercialized a new sandwich freezing device. The new device is inexpensive, portable and sterilizable. It can be used to rapid-freeze viruses, bacteria, yeast, cultured cells and animal and human tissues to a depth of 0.2 mm if tissues are prefixed with glutaraldehyde. The commercial availability of this device will expand application of rapid freezing to wide range of biological materials.

Many advances in PET/CT technology can potentially improve image quality and the ability to detect small lesions. A new digital TOF-PET/CT scanner based on silicon photomultipliers (SiPM) integrated with a Bayesian penalized likelihood (BPL) PET reconstruction algorithm (Q.Clear; GE Healthcare) has been introduced into clinical practice. The present study aimed to quantify the ability of a digital TOF-PET/CT scanner combined with BPL reconstruction to detect small lesions, and to determine the optimal penalization factor (β) in BPL to accurately detect such lesions. All PET data were acquired from a NEMA body phantom using a Discovery MI (DMI) PET/CT system (GE Healthcare). The phantom included six spheres with diameters of 4, 5, 6, 8, 10, and 13 mm, and contained a background activity level of 5.3 kBq/mL, with target-to-background ratios (TBR) of 4:1 and 8:1. Images were reconstructed using a baseline OSEM algorithm, with OSEM + PSF, OSEM + TOF, OSEM + PSF + TOF, and BPL + PSF + TOF (β: 50–400). The matrix size was 192 × 192 and 384 × 384. Data acquired in 100-min list mode were re-binned into acquisition times ranging from 2 to 100 min. The quantitative accuracy and detectability of small hot spheres were evaluated by physical assessment of a recovery coefficient (RC) and a detectability index (DI), as well as visual assessment of PET images at each acquisition time. The RC and DI of sub-centimeter spheres were improved, because the digital TOF-PET/CT scanner has a larger TOF performance gain due to better timing resolution. The RC and DI were higher with BPL in sub-centimeter spheres, than with other OSEM-based types of reconstruction. The BPL for an 8-mm sphere overestimated uptake due to edge artifact overshoot induced by PSF modeling. The variability of RC and DI for acquisition times and TBR differed considerably according to β values. The RC for ~ 8-mm spheres were > 1 at β values between 50 and 100, but were close to 1 at β value of 200. The visual scores for β = 200 in BPL were maximal, whereas those for spheres that were ≥ 6 mm exceeded the criterion of 3. The BPL in the digital TOF-PET/CT scanner improved the quantitation and detectability of sub-centimeter spheres compared with OSEM-based reconstruction. Optimization of the β value in BPL might allow the detection of lesions ≤ 6 mm, although detectability depended on the TBR of lesions. A β value of 200 seemed optimal for detecting sub-centimeter lesions.

This paper explores a theoretical account for depictive constructions in Japanese. Some previous analyses claim that depictive predicates in Japanese and their semantic DPs form a constituent, based on ellipsis phenomena. However, this paper provides several pieces of evidence showing that Japanese depictive predicates are independent of their semantic subjects. Therefore, an independent analysis should be applied to depictive constructions in Japanese. This paper argues that adjunction sites of depictive predicates are different, depending on the types of the predicates; object-oriented depictive predicates are adjoined to VP, and subject-oriented depictive predicates are either to vP or to TP. With this proposal, this paper demonstrates that all of the syntactic properties of depictive constructions can be accounted for, showing the plausibility of an independent constituent analysis of depictive constructions.

The uptake of cholesterol from the host is closely linked to the proliferation of pathogenic fungi and protozoa during infection. For some pathogenic fungi

Background Giant cell myocarditis (GCM) is a rare cause of fulminant heart failure (HF). The most common presentation is progressive hemodynamic deterioration, and a few cases present with idiopathic complete atrioventricular block (cAVB). The prognosis of GCM is poor, and GCM patients usually die of HF and ventricular arrhythmia unless cardiac transplantation is performed. Few reports have described the effects of treatments such as immunosuppression and detailed reverse remodelling in GCM patients. Case summary A 69-year-old female presented with cAVB. Transvenous pacemaker was implanted via the left subclavian vein. One and a half months later, she exhibited left ventricular dyssynchrony and lower left ventricular ejection fraction (LVEF), resulting in hospitalization for HF. She received cardiac resynchronization therapy; however, this had no apparently positive effects on her cardiac function. To investigate the cause of the lower LVEF, an endomyocardial biopsy was taken from the right ventricular septum. She was diagnosed with GCM and immediately received immunosuppression therapy with prednisolone and ciclosporin. This resulted in the functional recovery of the right ventricle; on the other hand, the left ventricle had still not recovered based on transthoracic echocardiography. Fortunately, she successfully recovered from severe HF without recurrence. Discussion This is a case of fulminant HF due to GCM which initially presented as cAVB. Moreover, this case demonstrates the quite difference of the functional recovery between the left ventricle and the right ventricle with immunosuppression therapy.

Chemical fixation has been used for observing the ultrastructure of cells and tissues. However, this method does not adequately preserve the ultrastructure of cells; artifacts and extraction of cell contents are usually observed. Rapid freezing is a better alternative for the preservation of cell structure. Sandwich freezing of living yeast or bacteria followed by freeze-substitution has been used for observing the exquisite natural ultrastructure of cells. Recently, sandwich freezing of glutaraldehyde-fixed cultured cells or human tissues has also been used to reveal the ultrastructure of cells and tissues. These studies have thus far been carried out with a handmade sandwich freezing device, and applications to studies in other laboratories have been limited. A new sandwich freezing device has recently been fabricated and is now commercially available. The present paper shows how to use the sandwich freezing device for rapid freezing of biological specimens, including bacteria, yeast, cultured cells, isolated cells, animal and human tissues, and viruses. Also shown is the preparation of specimens for ultrathin sectioning after rapid freezing and procedures for freeze-substitution, resin embedding, trimming of blocks, cutting of ultrathin sections, recovering of sections, staining, and covering of grids with support films.

Background The Bayesian penalized likelihood (BPL) algorithm Q.Clear (GE Healthcare) allows fully convergent iterative reconstruction that results in better image quality and quantitative accuracy, while limiting image noise. The present study aimed to optimize BPL reconstruction parameters for 18F-NaF PET/CT images and to determine the feasibility of 18F-NaF PET/CT image acquisition over shorter durations in clinical practice. Methods A custom-designed thoracic spine phantom consisting of several inserts, soft tissue, normal spine, and metastatic bone tumor, was scanned using a Discovery MI PET/CT scanner (GE Healthcare). The phantom allows optional adjustment of activity distribution, tumor size, and attenuation. We reconstructed PET images using OSEM + PSF + TOF (2 iterations, 17 subsets, and a 4-mm Gaussian filter), BPL + TOF (β = 200 to 700), and scan durations of 30–120 s. Signal-to-noise ratios (SNR), contrast, and coefficients of variance (CV) as image quality indicators were calculated, whereas the quantitative measures were recovery coefficients (RC) and RC linearity over a range of activity. We retrospectively analyzed images from five persons without bone metastases (male, n = 1; female, n = 4), then standardized uptake values (SUV), CV, and SNR at the 4th, 5th, and 6th thoracic vertebra were calculated in BPL + TOF (β = 400) images. Results The optimal reconstruction parameter of the BPL was β = 400 when images were acquired at 120 s/bed. At 90 s/bed, the BPL with a β value of 400 yielded 24% and 18% higher SNR and contrast, respectively, than OSEM (2 iterations; 120 s acquisitions). The BPL was superior to OSEM in terms of RC and the RC linearity over a range of activity, regardless of scan duration. The SUVmax were lower in BPL, than in OSEM. The CV and vertebral SNR in BPL were superior to those in OSEM. Conclusions The optimal reconstruction parameters of 18F-NaF PET/CT images acquired over different durations were determined. The BPL can reduce PET acquisition to 90 s/bed in 18F-NaF PET/CT imaging. Our results suggest that BPL (β = 400) on SiPM-based TOF PET/CT scanner maintained high image quality and quantitative accuracy even for shorter acquisition durations.

We previously reported that Candida albicans responded to mild heat stress in a range of temperature elevations simulating fever, and concluded that mild heat stress increases susceptibility to antifungal drugs. In this study, we show that mild heat stress causes a morphological change in hyphae during the process of biofilm formation. We found that mild heat stress extended the period of hyphal stage maintenance in C. albicans biofilm. Although the rate of hyphal change from yeast form to hyphal form reached the maximum within 3 hr, later, almost every cell quickly reverted to the yeast growth phase within 6 hr at 37°C but not at 39°C, or under mild heat stress. Electron microscopy using a smart specimen preparation technique revealed that mild heat stress significantly increased the thickness of the inner cell wall accompanied by a decrease in density of the outer cell wall in the hyphae of C. albicans biofilm. To identify the gene responsible for the morphological changes associated with mild heat stress, we performed microarray gene expression analysis. Eleven genes were upregulated and 17 genes were downregulated under mild heat stress in biofilm cells. The increased PHR1 gene expression in response to mild heat stress was confirmed in quantitative RT-PCR analysis. The mutant upregulated PHR1 expression showed the same sensitivity against antifungal drug micafungin as dependent on mild heat stress. Our findings point to possible therapeutic effects of hyperthermia as well as to the effect of fever during infections.

A calibration scheme of position and orientation errors of rotary axis average lines based on touch-trigger probing is widely available on many commercial five-axis machine tools. Such a measurement is influenced by error motions of both linear and rotary axes. This paper proposes a novel scheme to separately identify linear and rotary axis geometric errors by using a touch-trigger probe and an uncalibrated test piece. Whereas the proposed scheme is based on well-developed self-calibration schemes for the circularity measurement, an original contribution is that it is applied to separate linear and rotary axis geometric errors. Two case studies are presented. First, the proposed tests are performed on the same five-axis machine tool for half a year to observe a long-term change in linear and rotary axis geometric errors. The second case study investigates the influence of room temperature change on linear axis and rotary axis error motions.

We analyse the performance of a gas time projection chamber (TPC) as a high-performance gamma-ray telescope and polarimeter in the e$^+$e$^-$ pair creation regime. We use data collected at a gamma-ray beam of known polarisation. The TPC provides two orthogonal projections $(x,z)$ and $(y,z)$ of the tracks induced by each conversion in the gas volume. We use a simple vertex finder in which vertices and pseudo-tracks exiting from them are identified. We study the various contributions to the single-photon angular resolution using Monte Carlo simulations and compare them with the experimental data and find that they are in excellent agreement. The distribution of the azimutal angle of pair conversions shows a bias due to the non-cylindrical-symmetric structure of the detector. This bias would average out for a long duration exposure on a space mission, but for this pencil-beam characterisation we have ensured its accurate simulation by a double systematics control scheme, data taking with the detector rotated at several angles with respect to the beam polarisation direction and systematics control with a non-polarised beam. We measure, for the first time, the polarisation asymmetry of a linearly polarised gamma-ray beam in the low energy pair creation regime. This sub-GeV energy range is critical for cosmic sources as their spectra are power laws which fall quickly as a function of increasing energy. This work could pave the way to extending polarised gamma-ray astronomy beyond the MeV energy regime., Comment: Submitted to Astroparticle Physics

Tyrosine kinase inhibitors (TKI), including imatinib (IM), improve the outcome of CML therapy. However, TKI treatment is long-term and can induce resistance to TKI, which often leads to a poor clinical outcome in CML patients. Here, we examined the effect of continuous IM exposure on intracellular energy metabolism in K562 cells, a human Philadelphia chromosome-positive CML cell line, and its subsequent sensitivity to anti-cancer agents. Contrary to our expectations, we found that continuous IM exposure increased sensitivity to TKI. Cancer energy metabolism, characterized by abnormal glycolysis, is linked to cancer cell survival. Interestingly, glycolytic activity was suppressed by continuous exposure to IM, and autophagy increased to maintain cell viability by compensating for glycolytic suppression. Notably, increased sensitivity to TKI was not caused by glycolytic inhibition but by altered intracellular signaling, causing glycolytic suppression and increased autophagy, as evidenced by suppression of p70 S6 kinase 1 (S6K1) and activation of AMP-activated protein kinase (AMPK). Using another human CML cell line (KCL22 cells) and BCR/ABL+ Ba/F3 cells (mimicking Philadelphia chromosome-positive CML cells) confirmed that suppressing S6K1 and activating AMPK increased sensitivity to TKI. Furthermore, suppressing S6K1 and activating AMPK had a synergistic anti-cancer effect by inhibiting autophagy in the presence of TKI. The present study provides new insight into the importance of signaling pathways that affect cellular energy metabolism, and suggests that co-treatment with agents that disrupt energy metabolic signaling (using S6K1 suppressors and AMPK activators) plus blockade of autophagy may be strategies for TKI-based CML therapy.

// Shigeki Aoki 1, * , Michie Morita 1, * , Takuya Hirao 1 , Masashi Yamaguchi 2 , Reika Shiratori 1 , Megumi Kikuya 1 , Hiroji Chibana 2 and Kousei Ito 1 1 Laboratory of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba-city, Chiba 260-8675, Japan 2 Medical Mycology Research Center, Chiba University, Chiba-city, Chiba 260-8673, Japan * These authors have contributed equally to this work Correspondence to: Shigeki Aoki, email: aokishigeki@chiba-u.jp Keywords: ALL, autophagy, glucocorticoids, glycolysis, oxidative phosphorylation Received: June 16, 2017 Accepted: September 21, 2017 Published: October 09, 2017 ABSTRACT Acute lymphoblastic leukemia (ALL) is the most common childhood malignancy. Treatments include glucocorticoids (GCs) such as dexamethasone (Dex) and prednisolone, which may be of value when used alongside cytotoxic anti-cancer drugs. To predict therapeutic efficacy of GCs, their activity against ALL cells is usually examined prior to chemotherapy; however, few studies have examined their effects when used in combination with other drugs. The paradox is that cytotoxic anti-cancer drugs that are effective against proliferating cancer cells show synergistic effects when used with GCs that prevent cell proliferation. To address this point, we investigated intracellular energy metabolism in ALL CCRF-CEM cell clones classified according to their sensitivity to Dex and cytotoxic anti-cancer drugs in bulk cultures of mixed cells. We found that Dex suppressed glycolysis, the most important metabolic system in cancer cells, in cells that were damaged by etoposide (a cytotoxic anti-cancer drug), and the cells showed a concomitant increase in mitochondrial oxidative phosphorylation. Furthermore, autophagy, an intracellular bulk degradation system, regulated mitochondrial viability. We also found that mitochondria, whose function is enhanced by Dex, were susceptible to anti-cancer drugs that inhibit respiratory complexes (e.g., etoposide and daunorubicin), resulting in increased production of reactive oxygen species and subsequent cytotoxicity. Taken together, the present study points the way toward a more accurate prediction of the sensitivity of ALL cells to the combined action of anti-cancer drugs and GCs, by taking into consideration the shift in intracellular energy metabolism caused by GCs: namely, from glycolysis to mitochondrial oxidative phosphorylation mediated by autophagy.

In planarization processes of sapphire, lapping process takes a long time because sapphire is a hard material. In contrast, superfinishing, which involves fixed abrasive machining, can substitute for lapping, and it would be possible to shorten the amount of processing time. In this work, vitrified-bonded diamond superabrasive stones with different grain diameters are developed. Then, multistage superfinishing is investigated by combining these stones. Results indicate that the multistage process is capable of producing a 2 nmRa surface, equivalent of a lapped surface in less than 10 min. To improve the process of multistage superfinishing, a removal amount estimation method is developed based on the real contact pressure calculation. The working area ratio of the stone was calculated by considering elastic deformation during superfinishing. The contact ratio of sapphire is calculated considering the roughness of the pre-finished surface and grain depth of cut. Accordingly, the real contact pressure is calculated to estimate the removal amount during superfinishing and finished surface roughness was expected.

Current {\gamma}-ray telescopes based on photon conversions to electron-positron pairs, such as Fermi, use tungsten converters. They suffer of limited angular resolution at low energies, and their sensitivity drops below 1 GeV. The low multiple scattering in a gaseous detector gives access to higher angular resolution in the MeV-GeV range, and to the linear polarisation of the photons through the azimuthal angle of the electron-positron pair. HARPO is an R&D program to characterise the operation of a TPC (Time Projection Chamber) as a high angular-resolution and sensitivity telescope and polarimeter for {\gamma} rays from cosmic sources. It represents a first step towards a future space instrument. A 30 cm cubic TPC demonstrator was built, and filled with 2 bar argon-based gas. It was put in a polarised {\gamma}-ray beam at the NewSUBARU accelerator in Japan in November 2014. Data were taken at different photon energies from 1.7 MeV to 74 MeV, and with different polarisation configurations. The electronics setup is described, with an emphasis on the trigger system. The event reconstruction algorithm is quickly described, and preliminary measurements of the polarisation of 11 MeVphotons are shown., Comment: Proceedings VCI2016

Mast cells have secretory granules containing chemical mediators such as histamine and play important roles in the immune system. Polyamines are essential factors for cellular processes such as gene expression and translation. It has been reported that secretory granules contain both histamine and polyamines, which have similar chemical structures and are produced from the metabolism of cationic amino acids. We investigated the effect of polyamine depletion on mast cells using bone marrow-derived mast cells (BMMCs). Polyamine depletion was induced using α-difluoromethylornithine (DFMO), an irreversible inhibitor of ornithine decarboxylase. DFMO treatment resulted in a significant reduction of cell number and abnormal secretory granules in BMMCs. Moreover, the cells showed a 2.3-fold increase in intracellular histamine and up-regulation of histidine decarboxylase (HDC) at the transcriptional level during BMMC differentiation. Levels of the transcription factor kruppel-like factor 4 (KLF4) greatly decreased upon DFMO treatment; however, Klf4 mRNA was expressed at levels similar to controls. We determined the translational regulation of KLF4 using reporter genes encoding Klf4-luc2 fusion mRNA, for transfecting NIH3T3 cells, and performed in vitro translation. We found that the efficiency of KLF4 synthesis in response to DFMO treatment was enhanced by the existence of a GC-rich 5'-untranslated region (5'-UTR) on Klf4 mRNA, regardless of the recognition of the initiation codon. Taken together, these results indicate that the enhancement of histamine synthesis by DFMO depends on the up-regulation of Hdc expression, achieved by removal of transcriptional suppression of KLF4, during differentiation.

Most cancer cells rely on glycolysis to generate ATP, even when oxygen is available. However, merely inhibiting the glycolysis is insufficient for the eradication of cancer cells. One main reason for this is that cancer cells have the potential to adapt their metabolism to their environmental conditions. In this study, we investigated how cancer cells modify their intracellular metabolism when glycolysis is suppressed, using PANC-1 pancreatic cancer cells and two other solid tumor cell lines, A549 and HeLa. Our study revealed that glycolytically suppressed cells upregulated mitochondrial function and relied on oxidative phosphorylation (OXPHOS) to obtain the ATP necessary for their survival. Dynamic changes in intracellular metabolic profiles were also observed, reflected by the reduced levels of TCA cycle intermediates and elevated levels of most amino acids. Glutamine and glutamate were important for this metabolic reprogramming, as these were largely consumed by influx into the TCA cycle when the glycolytic pathway was suppressed. During the reprogramming process, activated autophagy was involved in modulating mitochondrial function. We conclude that upon glycolytic suppression in multiple types of tumor cells, intracellular energy metabolism is reprogrammed toward mitochondrial OXPHOS in an autophagy-dependent manner to ensure cellular survival.

Lysine-specific demethylases 1 and 2 (LSD1 and LSD2) are flavoenzyme demethylases, and their inhibitors are considered as potential chemical tools and anticancer agents. Here we report polyamine-based inhibitors of LSD1 and LSD2. In the initial screening, partially constrained polyamine 2 which contains three trans-cyclopentane units with a total of six stereogenic centers, showed the most potent LSD1-inhibitory activity. We then prepared a set of optical isomers of 2 and evaluated their inhibitory activities toward LSD1, LSD2, monoamine oxidases A and B (MAO-A and MAO-B). Optical isomers of 2 showed LSD1-inhibitory activity with Ki values of 2.2 to 6.4 μM, and LSD2-inhibitory activity with Ki values of 4.4 to 39 μM; there was a general preference for LSD1 to LSD2. All of them showed weak to negligible inhibition of MAO-A and MAO-B. This selectivity seemed to reflect the differences in the size and shape of the catalytic cavity of target enzymes, and our strategy of employing a set of optical isomers appears to be an effective approach for exploring the structural features of this family of enzymes. Polyamine 9 showed most potent LSD1-inhibitory activity (Ki = 2.2 μM in vitro), and it also inhibited the proliferation of HL-60 cells (IC50 = 49 μM). On the other hand, 12 was the most potent inhibitors of LSD2 with in vitro Ki values of 4.4 μM.

The present paper describes the applicability of the active flow control device, mini electromagnetic flap actuators attached on the leading edge of an airfoil, for the flow separation under both the steady and the unsteady flow conditions in the low Reynolds number region. At first, lift and drag have been measured for a wide variety of the wind speed Reynolds numbers and the angles of attack for the steady flow condition. Then, effects of some simple feedback flow controls, where the time-dependent signal of the lift-drag ratio have been used to detect the stall and served as a trigger to start the actuation, have been explored under the unsteady flow condition for evading the stall. In every low Reynolds number ranging from 30 000 to 80 000, the present actuators worked quite well to delay the stall, increasing in the lift and delaying the stall angle of attack. These aerodynamic modifications by the flap actuators obtained from the steady flow were found to be available even if the manipulation of the actuators started after the stall. Activation threshold of the lift-drag ratio as the input for the feedback control was determined from a stall classification map obtained under the steady flow experiment. Effectiveness of this feedback control was then demonstrated under the condition of the wind speed decrease (Reynolds number from 80 000 to 40 000) keeping the angle of attack constant at 11°, at which the stall occurs without the active control. Immediately after the sudden velocity decrease, the decrease in the lift-drag ratio were detected and the dynamic actuations were successfully started, resulting in evading the stall and keeping high and stable lift. An additional operation of the feedback, in which the running actuation is turned off when the lift-drag ratio shows lower than the second threshold value after operation, was revealed to be effective to keep the high lift force under the condition combined with the wind speed increase and decrease within the low Reynolds number range treated in this study.

Our basic cell biology research was aimed at investigating the effect on eukaryotic cells of the sudden loss of the F-actin cytoskeleton. Cells treated with latrunculin A (LA) in yeast extract peptone dextrose (YEPD) medium were examined using phase-contrast and fluorescent microscopy, freeze-substitution, transmission and scanning electron microscopy, counted using a Bürker chamber and their absorbance measured. The cells responded to the presence of LA, an F-actin inhibitor, with the disappearance of actin patches, actin cables and actin rings. This resulted in the formation of larger spherical cells with irregular morphology in the cell walls and ultrastructural disorder of the cell organelles and secretory vesicles. Instead of buds, LA-inhibited cells formed only 'table-mountain-like' wide flattened swellings without apical growth with a thinner glucan cell-wall layer containing β-1,3-glucan microfibrils. The LA-inhibited cells lysed. Actin cables and patches were required for bud formation and bud growth. In addition, actin patches were required for the formation of β-1,3-glucan microfibrils in the bud cell wall. LA has fungistatic, fungicidal and fungilytic effects on the budding yeast Saccharomyces cerevisiae.

Observing cells and cell components in three dimensions at high magnification in transmission electron microscopy requires preparing serial ultrathin sections of the specimen. Although preparing serial ultrathin sections is considered to be very difficult, it is rather easy if the proper method is used. In this paper, we show a step-by-step procedure for safely obtaining serial ultrathin sections of microorganisms. The key points of this method are: 1) to use the large part of the specimen and adjust the specimen surface and knife edge so that they are parallel to each other; 2) to cut serial sections in groups and avoid difficulty in separating sections using a pair of hair strands when retrieving a group of serial sections onto the slit grids; 3) to use a 'Section-holding loop' and avoid mixing up the order of the section groups; 4) to use a 'Water-surface-raising loop' and make sure the sections are positioned on the apex of the water and that they touch the grid first, in order to place them in the desired position on the grids; 5) to use the support film on an aluminum rack and make it easier to recover the sections on the grids and to avoid wrinkling of the support film; and 6) to use a staining tube and avoid accidentally breaking the support films with tweezers. This new method enables obtaining serial ultrathin sections without difficulty. The method makes it possible to analyze cell structures of microorganisms at high resolution in 3D, which cannot be achieved by using the automatic tape-collecting ultramicrotome method and serial block-face or focused ion beam scanning electron microscopy.

Tolvaptan (TLV) is a new vasopressin type 2 receptor antagonist effective in patients with heart failure (HF). We herein describe the case of an 84-year-old woman who developed acute renal injury induced by hypersensitivity to TLV. The patient had received an implanted pacemaker and was diagnosed with exacerbation of chronic HF due to atrial fibrillation, mitral regurgitation, tricuspid regurgitation and left ventricular dyssynchrony. Treatment with tolvaptan increased the urine volume, improved the dyspnea and decreased the edema. However, the patient's renal function and hyperkalemia worsened, and the blood eosinophil count increased without signs of dehydration or hypotension. Positive findings on a drug-induced lymphocyte stimulation test for TLV were consistent with this diagnosis.