Search results for: Yuta Nakano

Authors: Yuta Nakano, Kozo Kajiwara, Atsushi Hori, Takeshi Fujita

Abstract:

In association with the wave of electric vehicles (EV), low energy consumption systems have become more and more important. One of the key technologies to realize low energy consumption is a dynamic vision sensor (DVS), or we can call it an event sensor, neuromorphic vision sensor and so on. This sensor has several features, such as high temporal resolution, which can achieve 1 Mframe/s, and a high dynamic range (120 DB). However, the point that can contribute to low energy consumption the most is its sparsity; to be more specific, this sensor only captures the pixels that have intensity change. In other words, there is no signal in the area that does not have any intensity change. That is to say, this sensor is more energy efficient than conventional sensors such as RGB cameras because we can remove redundant data. On the other side of the advantages, it is difficult to handle the data because the data format is completely different from RGB image; for example, acquired signals are asynchronous and sparse, and each signal is composed of x-y coordinate, polarity (two values: +1 or -1) and time stamp, it does not include intensity such as RGB values. Therefore, as we cannot use existing algorithms straightforwardly, we have to design a new processing algorithm to cope with DVS data. In order to solve difficulties caused by data format differences, most of the prior arts make a frame data and feed it to deep learning such as Convolutional Neural Networks (CNN) for object detection and recognition purposes. However, even though we can feed the data, it is still difficult to achieve good performance due to a lack of intensity information. Although polarity is often used as intensity instead of RGB pixel value, it is apparent that polarity information is not rich enough. Considering this context, we proposed to use the timestamp information as a data representation that is fed to deep learning. Concretely, at first, we also make frame data divided by a certain time period, then give intensity value in response to the timestamp in each frame; for example, a high value is given on a recent signal. We expected that this data representation could capture the features, especially of moving objects, because timestamp represents the movement direction and speed. By using this proposal method, we made our own dataset by DVS fixed on a parked car to develop an application for a surveillance system that can detect persons around the car. We think DVS is one of the ideal sensors for surveillance purposes because this sensor can run for a long time with low energy consumption in a NOT dynamic situation. For comparison purposes, we reproduced state of the art method as a benchmark, which makes frames the same as us and feeds polarity information to CNN. Then, we measured the object detection performances of the benchmark and ours on the same dataset. As a result, our method achieved a maximum of 7 points greater than the benchmark in the F1 score.

Keywords: event camera, dynamic vision sensor, deep learning, data representation, object recognition, low energy consumption

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Authors: Osamu Takakuwa, Yuta Mano, Hitoshi Soyama

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This paper reveals the interaction between hydrogen and surface stress in austenitic stainless steel by X-ray diffraction stress measurement and thermal desorption analysis before and after being charged with hydrogen. The surface residual stress was varied by surface finishing using several disc polishing agents. The obtained results show that the residual stress near surface had a significant effect on hydrogen absorption behavior, that is, tensile residual stress promoted the hydrogen absorption and compressive one did opposite. Also, hydrogen induced equi-biaxial stress and this stress has a linear correlation with hydrogen content.

Keywords: hydrogen embrittlement, residual stress, surface finishing, stainless steel

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Authors: Yuta Moriyama, Etsuo Morishita

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Plasma actuator is very effective to recover stall flows over an upper airfoil surface. We first manufacture the actuator, test the stability of the device by trial and error basis and find the conditions for steady operations. We visualize the flow around an airfoil in the smoke tunnel and observe the stall recovery. The plasma actuator is stationary device and has no moving parts, and it might be an ideal device to control a model aircraft. We can use the actuator not only as a stall recovery device but also as a spoiler. We put the actuator near the leading edge of an elevator of a model aircraft as a spoiler, and measure the aerodynamic forces by a three-component balance. We observe the effect of the plasma actuator on the aerodynamic forces and the device effectiveness changes depending on the angle of attack whether it is positive or negative. We also visualize the flow caused by the plasma actuator by a desk-top Schlieren photography which is otherwise very difficult in a low-speed wind tunnel experiment.

Keywords: aerodynamics, plasma actuator, model aircraft, wind tunnel

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Authors: Yuta Tachibana, Ayuko Itsuki

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The three strains of bacteria (Lysinibacillus xylanilyticus, Bacillus kochii, and Enterococcus sp.) were isolated from the fermented Indigo (Polygonum tinctorium) dyeing solution using the dilution plate method and some fermentation conditions were determined. High-Performance Liquid Chromatography (HPLC) was used to determine the indigo concentration. When the isolated bacteria were cultured in the indigo liquid culture containing various sugars, starch, and ethanol, the indigo culture solutions containing galactose, mannose, ribose, and ethanol were remarkably decreased. Comparison of decreasing indigo between three strains showed that Enterococcus sp. had the fastest growth and decrease of indigo. However, decreasing indigo per unit micro biomass did not correspond to the results of decreasing indigo―Bacillus kochii had higher indigo-reducing activity than Enterococcus sp. and Lysinibacillus xylanilyticus.

Keywords: fermentation condition, high-performance liquid chromatography (HPLC), indigo dyeing solution, indigo-reducing activity

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Authors: Chanmoly Or, Kyuro Sasaki, Yuichi Sugai, Masanori Nakano, Motonao Imai

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Cold drainage mechanism of oil production is a complicated process which involves with solubility and foaming processes. Laboratory experiments were carried out to investigate the CO2 gas solubility in hexadecane (as light oil) and the effect of depressurization processes on microbubble generation. The experimental study of sensitivity parameters of temperature and pressure on CO2 gas solubility in hexadecane was conducted at temperature of 20 °C and 50 °C and pressure ranged 2.0–7.0 MPa by using PVT (RUSKA Model 2370) apparatus. The experiments of foamy hexadecane were also prepared by depressurizing from saturated pressure of 6.4 MPa and temperature of 50 °C. The experimental results show the CO2 gas solubility in hexadecane linearly increases with increasing pressure. At pressure 4.5 MPa, CO2 gas dissolved in hexadecane 2.5 mmol.g-1 for temperature of 50 °C and 3.5 mmol.g-1 for temperature of 20 °C. The bubbles of foamy hexadecane were observed that most of large bubbles were coalesced shortly whereas the small one keeps presence. The experimental result of foamy hexadecane indicated large depressurization step (∆P) produces high quality of foam with high microbubble distribution.

Keywords: CO2 gas solubility, depressurization process, foamy hexadecane, microbubble distribution

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Authors: D. Furuya, H. Ryujin, S. Takahira, Y. Sekine, Y. Oya, K. Sonoda, H. Ogawa, Y. Nomura, R. Maruyama, H. Kim, T. Kudo, A. Nakano, T. Saruta, S. Sugita, M. Nemoto, N. Tanahashi

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Purpose: To study the clinical characteristics of hypoglycemic coma in adult patients with type 1 or type 2 diabetes mellitus (DM). Methods: Participants in this retrospective study comprised 91 patients (54 men, 37 women; mean age ± standard deviation, 71.5 ± 12.6 years; range, 42-97 years) brought to our emergency department by ambulance with disturbance of consciousness in the 7 years from April 2007 to March 2014. Patients with hypoglycemia caused by alcoholic ketoacidosis, nutrition disorder, malignancies and psychological disorder were excluded. Results: Patients with type 1 (8 of 91) or type 2 DM (83 of 91) were analyzed. Mean blood sugar level was 31.6 ± 10.4 in all patients. A sulfonylurea (SU) was more commonly used in elderly (>75 years old; n=44)(70.5%) than in younger patients (36.2%, p < 0.05). Cases showing prolonged unconsciousness (range, 1 hour to 21 days; n=30) included many (p < 0.05) patients with dementia (13.3%; 0.5% without dementia) and fewer (p < 0.05) patients with type 1 DM (0%; 13.1% in type 2 DM). Specialists for DM (n=33) used SU less often (24.2%) than general physicians (69.0%, p < 0.05). Conclusion: In cases of hypoglycemic coma, SU was frequently used in elderly patients with DM.

Keywords: hypoglycemic coma, Diabetes mellitus, unconsciousness, elderly patients

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Authors: Yuta Moriyama, Tsuyoshi Yamazaki, Etsuo Morishita

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Several aerodynamic devices currently attract engineers and research students. The plasma actuator is one of them, and it is very effective to control the flow. The actuator recovers a separated flow to an attached one. The actuator is also inversely applied to a spoiler. The model aircraft might be controlled by this actuator. We develop a model aircraft with the plasma actuator. Another interesting device is the Wells turbine which rotates in one direction. The present authors propose a bicycle with the Wells turbine in the wheels. Power reduction is measured when the turbine is driven by an electric motor at the exit of a wind tunnel. Several Watts power reduction might be possible. This means that the torque of the bike can be augmented by the turbine in the cross wind. These devices are tested in the wind tunnel with a three-component balance and the aerodynamic forces and moment are obtained. In this paper, we introduce these devices and their aerodynamic characteristics. The control force and moment of the plasma actuator are clarified and the power reduction of the bicycle is quantified.

Keywords: aerodynamics, model aircraft, plasma actuator, Wells turbine

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Authors: Nur Syarafina Binti Othman, Tsubasa Jindo, Makato Yamada, Miho Tsuyama, Hitoshi Nakano

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A novel method to produce a fast high voltage solid states switch using Insulated Gate Bipolar Transistors (IGBTs) is presented for discharge-pumped gas lasers. The IGBTs are connected in series to achieve a high voltage rating. An avalanche transistor is used as the gate driver. The fast pulse generated by the avalanche transistor quickly charges the large input capacitance of the IGBT, resulting in a switch out of a fast high-voltage pulse. The switching characteristic of fast-high voltage solid state switch has been estimated in the multi-stage series-connected IGBT with the applied voltage of several tens of kV. Electrical circuit diagram and the mythology of fast-high voltage solid state switch as well as experimental results obtained are presented.

Keywords: high voltage, IGBT, solid state switch, bipolar transistor

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Authors: Kittipong Tripetch, Nobuhiko Nakano

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Two types of floating active resistors based on a complementary regulated cascode topology with cross-coupled regulated transistors are presented in this paper. The first topology is a high swing complementary regulated cascode active resistor. The second topology is a complementary common gate with a regulated cross coupled transistor. The small-signal input resistances of the floating resistors are derived. Three graphs of the input current versus the input voltage for different aspect ratios are designed and plotted using the Cadence Spectre 0.18-µm Rohm Semiconductor process. The total harmonic distortion graphs are plotted for three different aspect ratios with different input-voltage amplitudes and different input frequencies. From the simulation results, it is observed that a resistance of approximately 8.52 MΩ can be obtained from supply voltage at  ±0.9 V.

Keywords: floating active resistor, complementary common gate, complementary regulated cascode, current mirror

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Authors: Masashi Kanazawa, Takaaki Nakano, Masaaki Takemoto, Tomonori Imamura, Mamiko Sugimura, Toshitaka Ito

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Introduction: We experienced a massive disaster due to inhalation of water-soluble paint. Sixteen patients were brought to our emergency room, and pulmonary edema was revealed on the CT images of 12 cases. Purpose: Transition of chest CT-scan findings in cases with pulmonary edema was examined. Method: CT-scans were performed on the 1st, 2nd, 5th, and 19th days after the inhalation event. Patients whose pulmonary edema showed amelioration or exacerbation were classified into the improvement or the exacerbation group, respectively. Those with lung edema findings appearing at different sites after the second day were classified into the changing group. Results: Eight, one and three patients were in the improvement, exacerbation and changing groups, respectively. In all cases, the pulmonary edema had disappeared from CT images on the 19th day after the inhalation event. Conclusion: Inhalation of water-soluble paints is considered to be relatively safe. However, our observations in these emergency cases suggest that, even if pulmonary edema is not severe immediately after the exposure, new lesions may appear later and existing lesions may worsen. Follow-up imaging is thus necessary for about two weeks.

Keywords: CT scan, intoxication, pulmonary edema, water-soluble paint

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Authors: Mei Xutao, Nakano Kimihiko

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In order to enhance the energy harvesting efficiency, this paper presents a novel tri-stable energy harvesting system (TEHS), which is realized by the effect of magnetic force, in rotational motion to scavenge vibration energy. The device is meant to provide the power supply for wireless autonomous systems in low-frequency environment. The nonlinear TEHS is composed of the cantilever beam which is mounted on a rotating hub and partially covered by piezoelectric patch, a tip mass magnet in the end and two fixed magnets. A theoretical investigation using the Lagrangian formulation is derived to describe the motion of the energy harvesting system and the output voltage. Additionally, several numerical simulations were carried out to characterize the system under different external excitations and to validate its performance. The results demonstrated that TEHS owns a wide range of frequency of snap-through and high output voltage compared with the bi-stable energy harvesting system (BEHS). Moreover, some sets of experimental validations will be performed in the future work because the experimental setup is in the configuration now.

Keywords: piezoelectric beam, rotational motion, snap-through, tri-stable energy harvester

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Authors: Katsuyuki Umezawa, Makoto Nakazawa, Manabu Kobayashi, Yutaka Ishii, Michiko Nakano, Shigeichi Hirasawa

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This research aims to develop a self-study system equipped with an artificial teacher who gives advice to students by detecting the learners and to evaluate language learning in a unified framework. 'Detecting the learners' means that the system understands the learners' learning conditions, such as each learner’s degree of understanding, the difference in each learner’s thinking process, the degree of concentration or boredom in learning, and problem solving for each learner, which can be interpreted from learning behavior. In this paper, we propose a system to efficiently collect brain waves from learners by focusing on only the brain waves among the biological information for 'detecting the learners'. The conventional Electroencephalograph (EEG) measurement method during learning using a simple EEG has the following disadvantages. (1) The start and end of EEG measurement must be done manually by the experiment participant or staff. (2) Even when the EEG signal is weak, it may not be noticed, and the data may not be obtained. (3) Since the acquired EEG data is stored in each PC, there is a possibility that the time of data acquisition will be different in each PC. This time, we developed a system to collect brain wave data on the server side. This system overcame the above disadvantages.

Keywords: artificial teacher, e-learning, self-study system, simple EEG

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Authors: Keisuke Fujioka, Yuta Itoh, Masaru Minagawa, Shunji Kusayanagi

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In this study, building information models of the underground temporary structures and adjacent embedded pipes were constructed to show the importance of the information on underground pipes adjacent to the structures to enhance the productivity of execution of construction. Next, the bar chart used in actual construction process were employed to make the Gantt chart, and the critical pass analysis was carried out to show that accurate information on the arrangement of underground existing pipes can be used for the enhancement of the productivity of the construction of underground structures. In the analyzed project, significant construction delay was not caused by unforeseeable existence of underground pipes by the management ability of the construction manager. However, in many cases of construction executions in the developing countries, the existence of unforeseeable embedded pipes often causes substantial delay of construction. Design change based on uncertainty on the position information of embedded pipe can be also important risk for contractors in domestic construction. So CPM analyses were performed by a project-management-software to the situation that influence of the tasks causing construction delay was assumed more significant. Through the analyses, the efficiency of information management on underground pipes and BIM analysis in the design stage for workability improvement was indirectly confirmed.

Keywords: building-information modelling, construction information modelling, design improvement, infrastructure

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Authors: Kittipong Tripetch, Nobuhiko Nakano

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This paper proposes one stage and two stage CMOS Complementary Regulated Cascode Distributed Amplifier (CRCDA) design based on Inductive and Transformer coupling techniques. Usually, Distributed amplifier is based on inductor coupling between gate and gate of MOSFET and between drain and drain of MOSFET. But this paper propose some new idea, by coupling with differential primary windings of transformer between gate and gate of MOSFET first stage and second stage of regulated cascade amplifier and by coupling with differential secondary windings transformer of MOSFET between drain and drain of MOSFET first stage and second stage of regulated cascade amplifier. This paper also proposes polynomial modeling of Silicon Transformer passive equivalent circuit from Nanyang Technological University which is used to extract frequency response of transformer. Cadence simulation results are used to verify validity of transformer polynomial modeling which can be used to design distributed amplifier without Cadence. 4 parameters of scattering matrix of 2 port of the propose circuit is derived as a function of 4 parameters of impedance matrix.

Keywords: CMOS regulated cascode distributed amplifier, silicon transformer modeling with polynomial, low power consumption, distribute amplification technique

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Authors: Ryota Kuzuki, Mitsuhiro Kohara, Noboru Kizuki, Satoshi Yoshida, Hidetaka Hirai, Yuta Nezasa

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About fifty years have passed since Japan's gas supply industry became the first in the world to switch from coal and oil to LNG as a city gas feedstock. Since the Japanese government target of net-zero carbon emission in 2050 was announced in October 2020, it has now entered a new era of challenges to commit to the requirement for decarbonization. This paper describes the situation that synthetic methane, produced from renewable energy-derived hydrogen and recycled carbon, is a promising national policy of transition toward net-zero society. In November 2020, the Japan Gas Association announced the 'Carbon Neutral Challenge 2050' as a vision to contribute to the decarbonization of society by converting the city gas supply to carbon neutral. The key technologies is methanation. This paper shows that methanation is a realistic solution to contribute to the decarbonization of the whole country at a lower social cost, utilizing the supply chain that already exists, from LNG plants to burner chips. The challenges during the transition period (2030-2050), as CO2 captured from exhaust of thermal power plants and industrial factories are expected to be used, it is proposed that a system of guarantee of origin (GO) for H2 and CO2 should be established and harmonize international rules for calculating and allocating greenhouse gas emissions in the supply chain, a platform is also needed to manage tracking information on certified environmental values.

Keywords: synthetic methane, recycled carbon fuels, methanation, transition period, environmental value transfer platform

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Authors: Hiroaki Nasu, Ryota Miyamoto, Yuta Kodera, Yasuyuki Nogami

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To promote Industry 4.0 and Society 5.0 and so on, it is important to connect and share data so that every member can trust it. Blockchain (BC) technology is currently attracting attention as the most advanced tool and has been used in the financial field and so on. However, the data collaboration using BC has not progressed sufficiently among companies on the supply chain of manufacturing industry that handle sensitive data such as product quality, manufacturing conditions, etc. There are two main reasons why data utilization is not sufficiently advanced in the industrial supply chain. The first reason is that manufacturing information is top secret and a source for companies to generate profits. It is difficult to disclose data even between companies with transactions in the supply chain. In the blockchain mechanism such as Bitcoin using PKI (Public Key Infrastructure), in order to confirm the identity of the company that has sent the data, the plaintext must be shared between the companies. Another reason is that the merits (scenarios) of collaboration data between companies are not specifically specified in the industrial supply chain. For these problems this paper proposes a Business to Business (B2B) collaboration system using homomorphic encryption and BC technique. Using the proposed system, each company on the supply chain can exchange confidential information on encrypted data and utilize the data for their own business. In addition, this paper considers a scenario focusing on quality data, which was difficult to collaborate because it is a top secret. In this scenario, we show a implementation scheme and a benefit of concrete data collaboration by proposing a comparison protocol that can grasp the change in quality while hiding the numerical value of quality data.

Keywords: business to business data collaboration, industrial supply chain, blockchain, homomorphic encryption

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Authors: Yuta Chishima, Tatsuya Murakami, Michael McKay

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Temporal focus is described as one component of an individual’s time perspective and defined as the attention individuals devote to thinking about the past, present, and future. It affects how people incorporate perceptions about past experiences, current situations, and future expectations into their attitudes, cognitions, and behavior. The 12-item Temporal Focus Scale (TFS) is comprised of three-factors (past, current and future focus). The purpose of this study was to examine the reliability and validity of TFS scores in Japanese adolescents and young adults. The TFS was translated into Japanese by a professional translator, and the original author confirmed the back translated items. Study 1 involved 979 Japanese university students aged 18-25 years old in a questionnaire-based study. The hypothesized three-factor structure (with reliability) was confirmed, although there were problems with item 10. Internal consistency estimates for scores without item 10 were over .70, and test-retest reliability was also adequate. To verify the concurrent and convergent validity, we tested the relationship between TFS scores and life satisfaction, time perspective, self-esteem, and career efficacy. Results of correlational analyses supported our hypotheses. Specifically, future focus was strongly correlated to career efficacy, while past and current focus was not. Study 2 involved 1030 Japanese junior and junior high school students aged 12-18 years old in a questionnaire-based study, and results of multigroup analyses supported the age invariance of the TFS.

Keywords: Japanese, reliability, scale, temporal focus, validity

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Authors: Frank Jaksoni Mweta, Nozomu Adachi, Yoshikazu Todaka, Hirokazu Sato, Yuta Sato, Hiromi Miura, Masakazu Kobayashi, Chihiro Watanabe, Yoshiteru Aoyagi

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Advances in the consumption of hydrogen fuel increase demands of high strength steel pipes and storage tanks. However, high strength steels are highly sensitive to hydrogen embrittlement. Because the introduction of hydrogen into steel during the fabrication process or from the environment is unavoidable, it is essential to improve hydrogen embrittlement resistance of high strength steels through microstructural control. In the present study, the heterogeneous nanostructure with a tensile strength of about 1.8 GPa and the homogeneous nanostructure with a tensile strength of about 2.0 GPa of 316LN steels were generated after 92% heavy cold rolling and high-pressure torsion straining, respectively. The heterogeneous nanostructure is composed of twin domains, shear bands, and lamellar grains. The homogeneous nanostructure is composed of uniformly distributed ultrafine nanograins. The influence of heterogeneous and homogenous nanostructures on the hydrogen embrittlement resistance was investigated. The specimen for each nanostructure was electrochemically charged with hydrogen for 3, 6, 12, and 24 hours, respectively. Under the same hydrogen charging time, both nanostructures show almost the same concentration of the diffusible hydrogen based on the thermal desorption analysis. The tensile properties of the homogenous nanostructure were severely affected by the diffusible hydrogen. However, the diffusible hydrogen shows less impact on the tensile properties of the heterogeneous nanostructure. The difference in embrittlement behavior between the heterogeneous and homogeneous nanostructures was elucidated based on the mechanism of the cracks' growth observed in the tensile fractography. The hydrogen embrittlement was suppressed in the heterogeneous nanostructure because the twin domain became an obstacle for crack growth. The homogeneous nanostructure was not consisting an obstacle such as a twin domain; thus, the crack growth resistance was low in this nanostructure.

Keywords: diffusible hydrogen, heterogeneous nanostructure, homogeneous nanostructure, hydrogen embrittlement

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Authors: Haruka Ito, Toru Maruyama, Michihiro Ito, Chuya Shinzato, Hiroyuki Fujimura, Yoshikatsu Nakano, Shoichiro Suda, Sachiyo Aburatani, Haruko Takeyama

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Clarifying symbiotic relationships is one of the most important theme for understanding the marine eco-system. Coral reef has been regarded as an important environmental resource. Coral holobiont composed by coral, symbiotic microalgae zooxanthellae, and bacteria have complexed relationship. Zooxanthellae mainly supply organic matter to the host corals through their photosynthetic activity. The symbiotic relationship is indispensable for corals but may easily collapses due to the rise of seawater temperature. However, the molecular mechanism how seawater temperature influences their relationships still remain unclear. In this study, the transcriptomic analysis has applied to elucidate the coral-zooxanthellae relationships under high seawater temperature stress. To observe reactions of corals and zooxanthellae against the rise of seawater temperature, meta-gene expression in coral have been analyzed. The branches from six different colonies of a stony coral, Acropora tenuis, were sampled at nine times by 2016 at two locations, Ishikawabaru and South of Sesoko Island, Okinawa, Japan. The mRNAs extracted from the branches including zooxanthellae were sequenced by illumina HiSeq. Gene Set Enrichment Analysis (GSEA) based on hyper geometric distribution was performed. The seawater temperature at 2016 summer was unusually high, which was caused by El Niño event, and the number of zooxanthellae in coral was decreased in August. GSEA derived the several specific genes expressed in A. tenuis under heat stress conditions. The upregulated genes under heat stress highly related with infection immunity. The downregulated genes significantly contained cell cycle related genes. Thu, it is considered that heat stress cause disorder in cell metabolism of A. tenuis, resulting in serious influence to coral holobiont.

Keywords: coral, symbiosis, thermal stress response, transcriptome analysis

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Authors: Takaharu Hirai, Yuta Mitobe, Hiromi Hirai

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Introduction: Eating disorders (ED) are serious psychiatric disorders that affect individuals, especially adolescents. It has been suggested that nonclinical ED-like characteristics are related to sleep problems. However, studies exploring the association between potential ED and sleep disorders have primarily been conducted in Europe and the United States. We conducted a survey of Japanese adolescents to investigate this claim. Method: In this cross-sectional study, 398 school-aged adolescents, aged 12–18 years old, matched for gender ratio, responded to a self-administered questionnaire survey. We used the Eating Attitudes Test-26 (EAT-26) and the Athens Insomnia Scale (AIS) to measure potential ED and sleep problems, respectively. In this study, participants with an EAT-26 total score of 0–19 points were classified as non-ED, while those with scores of 20 points or higher were classified as potential ED. Result: Of the 398 participants, 17 (4.3%) had an EAT-26 total score of 20 or higher. Among boys, the rate was 6 of 199 participants (3%), and among girls, the rate was 11 of 182 participants (6%). There were 89 participants (22.4%) with an AIS score of 6 points or higher, of which 36 (17.6%) were boys, and 53 (27.5%) were girls. Adolescents with potential ED had significantly higher rates of daytime sleep problems than those without ED. Further, while examining the types of sleep problems, adolescents with potential ED had greater problems with a sense of well-being and physical and mental functioning during the day. In contrast, no significant associations were found between potential ED and sleep initiation, awakenings during the night, early morning awakening, total sleep duration, or overall quality of sleep. Finally, nocturnal and daytime sleep scores were significantly associated with dieting, bulimia, and oral control EAT-26 sub-scores. Discussion: While Japanese adolescents with possible ED do not experience nighttime sleep problems, they do experience problems related to well-being and mental and physical functioning, which are indicators of daytime sleep problems. This may assist with early detection of disordered eating in adolescents. The study suggested that professionals working towards adolescent mental health issues need an approach that comprehensively integrates both sleep problems and potential ED.

Keywords: adolescents, potential eating disorders, sleep problems, eating attitudes test-26

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Authors: Harunori Kawabe, Hideyuki Aoshima, Koji Murakami, Minoru Kawakami, Yuka Nakano, David D. Ordinario, C. W. Crawford, Iri Sato-Baran

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With the recent advances in gene editing technologies allowing the rewriting of genetic sequences, additional market growth in the global floriculture market beyond previous trends is anticipated through increasingly sophisticated plant breeding techniques. As a prerequisite for gene editing, the gene sequence of the target plant must first be identified. This necessitates the genetic analysis of plants with unknown gene sequences, the extraction of RNA, and comprehensive expression analysis. Consequently, a technology capable of consistently and effectively extracting high-purity DNA and RNA from plants is of paramount importance. Although model plants, such as Arabidopsis and tobacco, have established methods for DNA and RNA extraction, floricultural species such as roses present unique challenges. Different techniques to extract DNA and RNA from various floricultural species were investigated. Upon sampling and grinding the petals of several floricultural species, it was observed that nucleic acid extraction from the ground petal solutions of low viscosity was straightforward; solutions of high viscosity presented a significant challenge. It is postulated that the presence of substantial quantities of polysaccharides and polyphenols in the plant tissue was responsible for the inhibition of nucleic acid extraction. Consequently, attempts were made to extract high-purity DNA and RNA by improving the CTAB method and combining it with commercially available nucleic acid extraction kits. The quality of the total extracted DNA and RNA was evaluated using standard methods. Finally, the effectiveness of the extraction method was assessed by determining whether it was possible to create a library that could be applied as a suitable template for a next-generation sequencer. In conclusion, a method was developed for consistent and accurate nucleic acid extraction from high-viscosity floricultural samples. These results demonstrate improved techniques for DNA and RNA extraction from flowers, help facilitate gene editing of floricultural species and expand the boundaries of research and commercial opportunities.

Keywords: floriculture, gene editing, next-generation sequencing, nucleic acid extraction

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Authors: Akimitsu Kugimiya, Kouhei Iwato, Toru Saito, Jiro Kohda, Yasuhisa Nakano, Yu Takano

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The measurement of amino acid content is reported to be useful for the diagnosis of several types of diseases, including lung cancer, gastric cancer, colorectal cancer, breast cancer, prostate cancer, and diabetes. The conventional detection methods for amino acid are high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS), but they have several drawbacks as the equipment is cumbersome and the techniques are costly in terms of time and costs. In contrast, biosensors and biosensing methods provide more rapid and facile detection strategies that use simple equipment. The authors have reported a novel approach for the detection of each amino acid that involved the use of aminoacyl-tRNA synthetase (aaRS) as a molecular recognition element because aaRS is expected to a selective binding ability for corresponding amino acid. The consecutive enzymatic reactions used in this study are as follows: aaRS binds to its cognate amino acid and releases inorganic pyrophosphate. Hydrogen peroxide (H₂O₂) was produced by the enzyme reactions of inorganic pyrophosphatase and pyruvate oxidase. The Trinder’s reagent was added into the reaction mixture, and the absorbance change at 556 nm was measured using a microplate reader. In this study, an amino acid-sensing method using histidyl-tRNA synthetase (HisRS; histidine-specific aaRS) as molecular recognition element in combination with the Trinder’s reagent spectrophotometric method was developed. The quantitative performance and selectivity of the method were evaluated, and the optimal enzyme reaction and detection conditions were determined. The authors developed a simple and rapid method for detecting histidine with a combination of enzymatic reaction and spectrophotometric detection. In this study, HisRS was used to detect histidine, and the reaction and detection conditions were optimized for quantitation of these amino acids in the ranges of 1–100 µM histidine. The detection limits are sufficient to analyze these amino acids in biological fluids. This work was partly supported by Hiroshima City University Grant for Special Academic Research (General Studies).

Keywords: amino acid, aminoacyl-tRNA synthetase, biosensing, enzyme reaction

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Authors: Keigo Ide, Toru Maruyama, Michihiro Ito, Hiroyuki Fujimura, Yoshikatu Nakano, Shoichiro Suda, Sachiyo Aburatani, Haruko Takeyama

Abstract:

Understanding environmental system is one of the important tasks. In recent years, conservation of coral environments has been focused for biodiversity issues. The damage of coral reef under environmental impacts has been observed worldwide. However, the casual relationship between damage of coral and environmental impacts has not been clearly understood. On the other hand, structure/diversity of marine bacterial community may be relatively robust under the certain strength of environmental impact. To evaluate the coral environment conditions, it is necessary to investigate relationship between marine bacterial composition in coral reef and environmental factors. In this study, the Time Scale Network Analysis was developed and applied to analyze the marine environmental data for investigating the relationship among coral, bacterial community compositions and environmental factors. Seawater samples were collected fifteen times from November 2014 to May 2016 at two locations, Ishikawabaru and South of Sesoko in Sesoko Island, Okinawa. The physicochemical factors such as temperature, photosynthetic active radiation, dissolved oxygen, turbidity, pH, salinity, chlorophyll, dissolved organic matter and depth were measured at the coral reef area. Metagenome and metatranscriptome in seawater of coral reef were analyzed as the biological factors. Metagenome data was used to clarify marine bacterial community composition. In addition, functional gene composition was estimated from metatranscriptome. For speculating the relationships between physicochemical and biological factors, cross-correlation analysis was applied to time scale data. Even though cross-correlation coefficients usually include the time precedence information, it also included indirect interactions between the variables. To elucidate the direct regulations between both factors, partial correlation coefficients were combined with cross correlation. This analysis was performed against all parameters such as the bacterial composition, the functional gene composition and the physicochemical factors. As the results, time scale network analysis revealed the direct regulation of seawater temperature by photosynthetic active radiation. In addition, concentration of dissolved oxygen regulated the value of chlorophyll. Some reasonable regulatory relationships between environmental factors indicate some part of mechanisms in coral reef area.

Keywords: coral environment, marine microbiology, network analysis, omics data analysis

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Authors: Yuta Kurashina, Chikahiro Imashiro, Kiyoshi Ohnuma, Kenjiro Takemura

Abstract:

Research objectives and goals: To realize clinical applications of regenerative medicine, a mass cell culture is highly required. In a conventional cell culture, trypsinization was employed for cell detachment. However, trypsinization causes proliferation decrease due to injury of cell membrane. In order to detach cells using an enzyme-free method, therefore, this study proposes a novel cell detachment method capable of detaching adherent cells using acoustic radiation pressure exposed to the dish by the assistance of serum-free medium with ITS liquid medium supplement. Methods used In order to generate acoustic radiation pressure, a piezoelectric ceramic plate was glued on a glass plate to configure an ultrasonic transducer. The glass plate and a chamber wall compose a chamber in which a culture dish is placed in glycerol. Glycerol transmits acoustic radiation pressure to adhered cells on the culture dish. To excite a resonance vibration of transducer, AC signal with 29-31 kHz (swept) and 150, 300, and 450 V was input to the transducer for 5 min. As a pretreatment to reduce cell adhesivity, serum-free medium with ITS liquid medium supplement was spread to the culture dish before exposed to acoustic radiation pressure. To evaluate the proposed cell detachment method, C2C12 myoblast cells (8.0 × 104 cells) were cultured on a ø35 culture dish for 48 hr, and then the medium was replaced with the serum-free medium with ITS liquid medium supplement for 24 hr. We replaced the medium with phosphate buffered saline and incubated cells for 10 min. After that, cells were exposed to the acoustic radiation pressure for 5 min. We also collected cells by using trypsinization as control. Cells collected by the proposed method and trypsinization were respectively reseeded in ø60 culture dishes and cultured for 24 hr. Then, the number of proliferated cells was counted. Results achieved: By a phase contrast microscope imaging, shrink of lamellipodia was observed before exposed to acoustic radiation pressure, and no cells remained on the culture dish after the exposed of acoustic radiation pressure. This result suggests that serum-free medium with ITS liquid inhibits adhesivity of cells and acoustic radiation pressure detaches cells from the dish. Moreover, the number of proliferated cells 24 hr after collected by the proposed method with 150 and 300 V is the same or more than that by trypsinization, i.e., cells were proliferated 15% higher with the proposed method using acoustic radiation pressure than with the traditional cell collecting method of trypsinization. These results proved that cells were able to be collected by using the appropriate exposure of acoustic radiation pressure. Conclusions: This study proposed a cell detachment method using acoustic radiation pressure by the assistance of serum-free medium. The proposed method provides an enzyme-free cell detachment method so that it may be used in future clinical applications instead of trypsinization.

Keywords: acoustic radiation pressure, cell detachment, enzyme free, ultrasonic transducer

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Authors: Misa Nakao, Yuta Kurashina, Chikahiro Imashiro, Kenjiro Takemura

Abstract:

The Research Goal: With the growing demand for regenerative medicine, an effective mass cell culture process is required. In a repetitive subculture process for proliferating cells, preparing single cell suspension which does not contain any cell aggregates is highly required because cell aggregates often raise various undesirable phenomena, e.g., apoptosis and decrease of cell proliferation. Since cell aggregates often occur in cell suspension during conventional subculture processes, this study proposes a single cell sorter driven by a resonance vibration of a cell culture substrate. The Method and the Result: The single cell sorter is simply composed of a cell culture substrate and a glass pipe vertically placed against the cell culture substrate with a certain gap corresponding to a cell diameter. The cell culture substrate is made of biocompatible stainless steel with a piezoelectric ceramic disk glued to the bottom side. Applying AC voltage to the piezoelectric ceramic disk, an out-of-plane resonance vibration with a single nodal circle of the cell culture substrate can be excited at 5.5 kHz. By doing so, acoustic radiation force is emitted, and then cell suspension containing only single cells is pumped into the pipe and collected. This single cell sorter is effective to collect single cells selectively in spite of its quite simple structure. We collected C2C12 myoblast cell suspension by the single cell sorter with the vibration amplitude of 12 µmp-p and evaluated the ratio of single cells in number against the entire cells in the suspension. Additionally, we cultured the collected cells for 72 hrs and measured the number of cells after the cultivation in order to evaluate their proliferation. As a control sample, we also collected cell suspension by conventional pipetting, and evaluated the ratio of single cells and the number of cells after the 72-hour cultivation. The ratio of single cells in the cell suspension collected by the single cell sorter was 98.2%. This ratio was 9.6% higher than that collected by conventional pipetting (statistically significant). Moreover, the number of cells cultured for 72 hrs after the collection by the single cell sorter yielded statistically more cells than that collected by pipetting, resulting in a 13.6% increase in proliferated cells. These results suggest that the cell suspension collected by the single cell sorter driven by the resonance vibration hardly contains cell aggregates whose diameter is larger than the gap between the cell culture substrate and the pipe. Consequently, the cell suspension collected by the single cell sorter maintains high cell proliferation. Conclusions: In this study, we developed a single cell sorter capable of sorting and pumping single cells by a resonance vibration of a cell culture substrate. The experimental results show the single cell sorter collects single cell suspension which hardly contains cell aggregates. Furthermore, the collected cells show higher proliferation than that of cells collected by conventional pipetting. This means the resonance vibration of the cell culture substrate can benefit us with the increase in efficiency of mass cell culture process for clinical applications.

Keywords: acoustic radiation force, cell proliferation, regenerative medicine, resonance vibration, single cell sorter

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Authors: Natsuki Kishizawa, Keiko Nakano, Hussam Organji, Amer Shaiban, Mohammad Albeirutty

Abstract:

One of the most important tasks in operating desalination plants using a reverse osmosis (RO) method is preventing RO membrane fouling caused by foulants found in seawater. Optimal design of the pre-treatment process of RO process for plants enables the reduction of foulants. Therefore, a quantitative evaluation of the fouling risk in pre-treated water, which is fed to RO, is required for optimal design. Some measurement methods for water quality such as silt density index (SDI) and total organic carbon (TOC) have been conservatively applied for evaluations. However, these methods have not been effective in some situations for evaluating the fouling risk of RO feed water. Furthermore, stable management of plants will be possible by alerts and appropriate control of the pre-treatment process by using the method if it can be applied to the inline monitoring system for the fouling risk of RO feed water. The purpose of this study is to develop a method to evaluate the fouling risk of RO feed water. We applied a quartz crystal microbalance (QCM) to measure the amount of foulants found in seawater using a sensor whose surface is coated with polyamide thin film, which is the main material of a RO membrane. The increase of the weight of the sensor after a certain length of time in which the sample water passes indicates the fouling risk of the sample directly. We classified the values as “FP: Fouling Potential”. The characteristics of the method are to measure the very small amount of substances in seawater in a short time: < 2h, and from a small volume of the sample water: < 50mL. Using some RO cell filtration units, a higher correlation between the pressure increase given by RO fouling and the FP from the method than SDI and TOC was confirmed in the laboratory-scale test. Then, to establish the correlation in the actual bench-scale RO membrane module, and to confirm the feasibility of the monitoring system as a control tool for the pre-treatment process, we have started a long-term test at an experimental desalination site by the Red Sea in Jeddah, Kingdom of Saudi Arabia. Implementing inline equipment for the method made it possible to measure FP intermittently (4 times per day) and automatically. Moreover, for two 3-month long operations, the RO operation pressure among feed water samples of different qualities was compared. The pressure increase through a RO membrane module was observed at a high FP RO unit in which feed water was treated by a cartridge filter only. On the other hand, the pressure increase was not observed at a low FP RO unit in which feed water was treated by an ultra-filter during the operation. Therefore, the correlation in an actual scale RO membrane was established in two runs of two types of feed water. The result suggested that the FP method enables the evaluation of the fouling risk of RO feed water.

Keywords: fouling, monitoring, QCM, water quality

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Authors: Sundaram Arulmozhiraja, Kanade Shimizu, Yuta Yamamoto, Satoshi Ichikawa, Maenaka Katsumi, Hiroaki Tokiwa

Abstract:

Drug discovery is shifting from small molecule based drugs targeting local active site to middle molecules (MM) targeting large, flat, and groove-shaped binding sites, for example, protein-protein interface because at least half of all targets assumed to be involved in human disease have been classified as “difficult to drug” with traditional small molecules. Hence, MMs such as peptides, natural products, glycans, nucleic acids with various high potent bioactivities become important targets for drug discovery programs in the recent years as they could be used for ‘undruggable” intracellular targets. Cell membrane permeability is one of the key properties of pharmacodynamically active MM drug compounds and so evaluating this property for the potential MMs is crucial. Computational prediction for cell membrane permeability of molecules is very challenging; however, recent advancement in the molecular dynamics simulations help to solve this issue partially. It is expected that MMs with high membrane permeability will enable drug discovery research to expand its borders towards intracellular targets. Further to understand the chemistry behind the permeability of MMs, it is necessary to investigate their conformational changes during the permeation through membrane and for that their interactions with the membrane field should be studied reliably because these interactions involve various non-bonding interactions such as hydrogen bonding, -stacking, charge-transfer, polarization dispersion, and non-classical weak hydrogen bonding. Therefore, parameters-based classical mechanics calculations are hardly sufficient to investigate these interactions rather, quantum mechanical (QM) calculations are essential. Fragment molecular orbital (FMO) method could be used for such purpose as it performs ab initio QM calculations by dividing the system into fragments. The present work is aimed to study the cell permeability of middle molecules using molecular dynamics simulations and FMO-QM calculations. For this purpose, a natural compound syringolin and its analogues were considered in this study. Molecular simulations were performed using NAMD and Gromacs programs with CHARMM force field. FMO calculations were performed using the PAICS program at the correlated Resolution-of-Identity second-order Moller Plesset (RI-MP2) level with the cc-pVDZ basis set. The simulations clearly show that while syringolin could not permeate the membrane, its selected analogues go through the medium in nano second scale. These correlates well with the existing experimental evidences that these syringolin analogues are membrane-permeable compounds. Further analyses indicate that intramolecular -stacking interactions in the syringolin analogues influenced their permeability positively. These intramolecular interactions reduce the polarity of these analogues so that they could permeate the lipophilic cell membrane. Conclusively, the cell membrane permeability of various middle molecules with potent bioactivities is efficiently studied using molecular dynamics simulations. Insight of this behavior is thoroughly investigated using FMO-QM calculations. Results obtained in the present study indicate that non-bonding intramolecular interactions such as hydrogen-bonding and -stacking along with the conformational flexibility of MMs are essential for amicable membrane permeation. These results are interesting and are nice example for this theoretical calculation approach that could be used to study the permeability of other middle molecules. This work was supported by Japan Agency for Medical Research and Development (AMED) under Grant Number 18ae0101047.

Keywords: fragment molecular orbital theory, membrane permeability, middle molecules, molecular dynamics simulation

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