Search results for: Keiko Sakai
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 31

Search results for: Keiko Sakai

31 The Concentration of Formaldehyde in Rainwater and Typhoon Rainwater at Sakai City, Japan

Authors: Chinh Nguyen Nhu Bao, Hien To Thi, Norimichi Takenaka

Abstract:

Formaldehyde (HCHO) concentrations in rainwater including in tropical storms in Sakai City, Osaka, Japan have been measured continuously during rain event by developed chemiluminescence method. The level of formaldehyde was ranged from 15 µg/L to 500 µg/L. The high concentration of HCHO in rainwater is related to the wind direction from the south and west sides of Sakai City where manufactures related to chemicals, oil-refinery, and steel. The in-situ irradiated experiment on rainwater sample was conducted to prove the aqueous phase photo-production of HCHO and the degradation of HCHO. In the daytime, the aqueous phase photolysis is the source of HCHO in rainwater (4.52 ± 5.74 µg/L/h for UV light source in-situ condition, 2.84-8.96 µg/L/h under sunlight). However, in the night time, the degradation is the function of microorganism.

Keywords: chemiluminescence, formaldehyde, rainwater, typhoon

Procedia PDF Downloads 158
30 Smart Container Farming: Innovative Urban Strawberry Farming Model from Japan to the World

Authors: Nishantha Giguruwa

Abstract:

This research investigates the transformative potential of smart container farming, building upon the successful cultivation of Japanese mushrooms at Sakai Farms in Aichi Prefecture, Japan, under the strategic collaboration with the Daikei Group. Inspired by this success, the study focuses on establishing an advanced urban strawberry farming laboratory with the aim of understanding strawberry farming technologies, fostering collaboration, and strategizing marketing approaches for both local and global markets. Positioned within the business framework of Sakai Farms and the Daikei Group, the study underscores the sustainability and forward-looking solutions offered by smart container farming in agriculture. The global significance of strawberries is emphasized, acknowledging their economic and cultural importance. The detailed examination of strawberry farming intricacies informs the technological framework developed for smart containers, implemented at Sakai Farms. Integral to this research is the incorporation of controlled bee pollination, a groundbreaking addition to the smart container farming model. The study anticipates future trends, outlining avenues for continuing exploration, stakeholder collaborations, policy considerations, and expansion strategies. Notably, the author expresses a strategic intent to approach the global market, leveraging the foreign student/faculty base at Ritsumeikan Asia Pacific University, where the author is affiliated. This unique approach aims to disseminate the research findings globally, contributing to the broader landscape of agricultural innovation. The integration of controlled bee pollination within this innovative framework not only enhances sustainability but also marks a significant stride in the evolution of urban agriculture, aligning with global agricultural trends.

Keywords: smart container farming, urban agriculture, strawberry farming technologies, controlled bee pollination, agricultural innovation

Procedia PDF Downloads 48
29 Tardiness and Self-Regulation: Degree and Reason for Tardiness in Undergraduate Students in Japan

Authors: Keiko Sakai

Abstract:

In Japan, all stages of public education aim to foster a zest for life. ‘Zest’ implies solving problems by oneself, using acquired knowledge and skills. It is related to the self-regulation of metacognition. To enhance this, establishing good learning habits is important. Tardiness in undergraduate students should be examined based on self-regulation. Accordingly, we focussed on self-monitoring and self-planning strategies among self-regulated learning factors to examine the causes of tardiness. This study examines the impact of self-monitoring and self-planning learning skills on the degree and reason for tardiness in undergraduate students. A questionnaire survey was conducted, targeted to undergraduate students in University X in the autumn semester of 2018. Participants were 247 (average age 19.7, SD 1.9; 144 males, 101 females, 2 no answers). The survey contained the following items and measures: school year, the number of classes in the semester, degree of tardiness in the semester (subjective degree and objective times), active participation in and action toward schoolwork, self-planning and self-monitoring learning skills, and reason for tardiness (open-ended question). First, the relation between strategies and tardiness was examined by multiple regressions. A statistically significant relationship between a self-monitoring learning strategy and the degree of subjective and objective tardiness was revealed, after statistically controlling the school year and the number of classes. There was no significant relationship between a self-planning learning strategy and the degree of tardiness. These results suggest that self-monitoring skills reduce tardiness. Secondly, the relation between a self-monitoring learning strategy and the reason of tardiness was analysed, after classifying the reason for tardiness into one of seven categories: ‘overslept’, ‘illness’, ‘poor time management’, ‘traffic delays’, ‘carelessness’, ‘low motivation’, and ‘stuff to do’. Chi-square tests and Fisher’s exact tests showed a statistically significant relationship between a self-monitoring learning strategy and the frequency of ‘traffic delays’. This result implies that self-monitoring skills prevent tardiness because of traffic delays. Furthermore, there was a weak relationship between a self-monitoring learning strategy score and the reason-for-tardiness categories. When self-monitoring skill is higher, a decrease in ‘overslept’ and ‘illness’, and an increase in ‘poor time management’, ‘carelessness’, and ‘low motivation’ are indicated. It is suggested that a self-monitoring learning strategy is related to an internal causal attribution of failure and self-management for how to prevent tardiness. From these findings, the effectiveness of a self-monitoring learning skill strategy for reducing tardiness in undergraduate students is indicated.

Keywords: higher-education, self-monitoring, self-regulation, tardiness

Procedia PDF Downloads 132
28 In-situ Observations Using SEM-EBSD for Bending Deformation in Single-Crystal Materials

Authors: Yuko Matayoshi, Takashi Sakai, Yin-Gjum Jin, Jun-ichi Koyama

Abstract:

To elucidate the material characteristics of single crystals of pure aluminum and copper, the respective relations between crystallographic orientations and micro structures were examined, along with bending and mechanical properties. The texture distribution was also analysed. Bending tests were performed in a SEM apparatus while its behaviors were observed. Some analytical results related to crystal direction maps, inverse pole figures, and textures were obtained from electron back scatter diffraction (EBSD) analyses.

Keywords: pure aluminum, pure copper, single crystal, bending, SEM-EBSD analysis, texture, microstructure

Procedia PDF Downloads 363
27 Texture Observation of Bending by XRD and EBSD Method

Authors: Takashi Sakai, Yuri Shimomura

Abstract:

The crystal orientation is a factor that affects the microscopic material properties. Crystal orientation determines the anisotropy of the polycrystalline material. And it is closely related to the mechanical properties of the material. In this paper, for pure copper polycrystalline material, two different methods; X-Ray Diffraction (XRD) and Electron Backscatter Diffraction (EBSD); and the crystal orientation were analyzed. In the latter method, it is possible that the X-ray beam diameter is thicker as compared to the former, to measure the crystal orientation macroscopically relatively. By measurement of the above, we investigated the change in crystal orientation and internal tissues of pure copper.

Keywords: bending, electron backscatter diffraction, X-ray diffraction, microstructure, IPF map, orientation distribution function

Procedia PDF Downloads 320
26 Emergency Condition Discrimination for Single People Using a CO2 Sensor and Body Detectors

Authors: Taiyo Matsumura, Kota Funabashi, Nobumichi Sakai, Takashi Ono

Abstract:

The purpose of this research is to construct a watching system that monitors human activity in a room and detects abnormalities at an early stage to prevent unattended deaths of people living alone. In this article, we propose a method whereby highly urgent abnormal conditions of a person are determined by changes in the concentration of CO2 generated from activity and respiration in a room. We also discussed the effects the amount of activity has on the determination. The results showed that this discrimination method is not dependent on the amount of activity and is effective in judging highly urgent abnormal conditions.

Keywords: abnormal conditions, multiple sensors, people living alone, respiratory arrest, unattended death, watching system

Procedia PDF Downloads 134
25 SEM-EBSD Observation for Microtubes by Using Dieless Drawing Process

Authors: Takashi Sakai, Itaru Kumisawa

Abstract:

Because die drawing requires insertion of a die, a plug, or a mandrel, higher precision and efficiency are demanded for drawing equipment for a tube having smaller diameter. Manufacturing of such tubes is also accompanied by problems such as cracking and fracture. We specifically examine dieless drawing, which is less affected by these drawing-related difficulties. This deformation process is governed by a similar principle to that of reduction in diameter when pulling a heated glass tube. We conducted dieless drawing of SUS304 stainless steel microtubes under various conditions with three factor parameters of heating temperature, area reduction, and drawing speed. We used SEM-EBSD to observe the processing condition effects on microstructural elements. As the result of this study, crystallographic orientation of microtube is clear by using SEM-EBSD analysis.

Keywords: microtube, dieless drawing, IPF (inverse pole figure), GOS (grain orientation spread), crystallographic analysis

Procedia PDF Downloads 243
24 LaPEA: Language for Preprocessing of Edge Applications in Smart Factory

Authors: Masaki Sakai, Tsuyoshi Nakajima, Kazuya Takahashi

Abstract:

In order to improve the productivity of a factory, it is often the case to create an inference model by collecting and analyzing operational data off-line and then to develop an edge application (EAP) that evaluates the quality of the products or diagnoses machine faults in real-time. To accelerate this development cycle, an edge application framework for the smart factory is proposed, which enables to create and modify EAPs based on prepared inference models. In the framework, the preprocessing component is the key part to make it work. This paper proposes a language for preprocessing of edge applications, called LaPEA, which can flexibly process several sensor data from machines into explanatory variables for an inference model, and proves that it meets the requirements for the preprocessing.

Keywords: edge application framework, edgecross, preprocessing language, smart factory

Procedia PDF Downloads 137
23 Study on the Suppression of Hydrogen Generation by Aluminum-Containing Waste Incineration Ash and Water

Authors: Hideyuki Onodera, Ryoji Imai, Masahiro Sakai

Abstract:

Explosions have occurred in incineration plants in conveyors, ash pits, and other locations. The cause of such explosions is thought to be the reaction of metallic aluminum contained in the ash with water used to cool the ash and prevent scattering, resulting in the generation of hydrogen. Given this background, conveyors and other equipment have been damaged by explosions, which has hindered the stable operation of incineration plants. In addition, workers may be injured by equipment explosions, creating an unsafe situation. To remedy these problems, it is necessary to devise a way to prevent hydrogen explosions from occurring. To overcome this problem, we conducted a hydrogen generation reaction experiment using simulated incinerator ash powder containing aluminum, calcium oxide, and water and confirmed that conditions exist to stop the hydrogen generation reaction. The results of this research may contribute to the suppression of hydrogen explosions at incineration plants.

Keywords: waste incinerated ash, aluminum, water, hydrogen, suppression of hydrogen generation, incineration plant

Procedia PDF Downloads 23
22 BECOME: Body Experience-Based Co-Operation between Juveniles through Mutually Excited Team Gameplay

Authors: Tsugunosuke Sakai, Haruya Tamaki, Ryuichi Yoshida, Ryohei Egusa, Etsuji Yamaguchi, Shigenori Inagaki, Fusako Kusunoki, Miki Namatame, Masanori Sugimoto, Hiroshi Mizoguchi

Abstract:

We aim to develop a full-body interaction game that could let children cooperate and interact with other children in small groups. As the first step for our aim, the objective of the full-body interaction game developed in this study is to make interaction between children. The game requires two children to jump together with the same timing. We let children experience the game and answer the questionnaires. The children using several strategies to coordinate the timing of their jumps were observed. These included shouting time, watching each other, and jumping in a constant rhythm as if they were skipping rope. In this manner, we observed the children playing the game while cooperating with each other. The results of a questionnaire to evaluate the proposed interactive game indicate that the jumping game was a very enjoyable experience in which the participants could immerse themselves. Therefore, the game enabled children to experience cooperation with others by using body movements.

Keywords: children, cooperation, full-body interaction game, kinect sensor

Procedia PDF Downloads 365
21 Identity-Based Encryption: A Comparison of Leading Classical and Post-Quantum Implementations in an Enterprise Setting

Authors: Emily Stamm, Neil Smyth, Elizabeth O'Sullivan

Abstract:

In Identity-Based Encryption (IBE), an identity, such as a username, email address, or domain name, acts as the public key. IBE consolidates the PKI by eliminating the repetitive process of requesting public keys for each message encryption. Two of the most popular schemes are Sakai-Kasahara (SAKKE), which is based on elliptic curve pairings, and the Ducas, Lyubashevsky, and Prest lattice scheme (DLP- Lattice), which is based on quantum-secure lattice cryptography. In or- der to embed the schemes in a standard enterprise setting, both schemes are implemented as shared system libraries and integrated into a REST service that functions at the enterprise level. The performance of both schemes as libraries and services is compared, and the practicalities of implementation and application are discussed. Our performance results indicate that although SAKKE has the smaller key and ciphertext sizes, DLP-Lattice is significantly faster overall and we recommend it for most enterprise use cases.

Keywords: identity-based encryption, post-quantum cryptography, lattice-based cryptography, IBE

Procedia PDF Downloads 120
20 The Intervention Effect of Gratitude Skills Training on the Reduction of Loneliness

Authors: T. Sakai, A. Aikawa

Abstract:

This study defined 'gratitude skills training' as a social skills training which would become a new intervention method about gratitude intervention. The purpose of this study was to confirm the intervention effect of gratitude skills training on the reduction of loneliness. The participants in this study were university students (n = 36). A waiting list control design was used, in which the participants were assigned either to a training group (n = 18) or a waiting list control group (n = 18); the latter group took the same training after the first group had been trained. The two-week gratitude skills training comprised of three sessions (50 minutes per each of sessions). In the three sessions, the guidebook and the homework developed in this study were used. Results showed that gratitude skills training improved the participants’ gratitude skills. The results also indicated the intervention effect of gratitude skills training on the reduction of loneliness during the follow-up after three weeks. This study suggests that gratitude skills training can reduce loneliness. The gratitude skills training has a possibility of becoming a new treatment to reduce loneliness.

Keywords: gratitude skills, loneliness, social skills training, well-being

Procedia PDF Downloads 193
19 Analysis of Sediment Distribution around Karang Sela Coral Reef Using Multibeam Backscatter

Authors: Razak Zakariya, Fazliana Mustajap, Lenny Sharinee Sakai

Abstract:

A sediment map is quite important in the marine environment. The sediment itself contains thousands of information that can be used for other research. This study was conducted by using a multibeam echo sounder Reson T20 on 15 August 2020 at the Karang Sela (coral reef area) at Pulau Bidong. The study aims to identify the sediment type around the coral reef by using bathymetry and backscatter data. The sediment in the study area was collected as ground truthing data to verify the classification of the seabed. A dry sieving method was used to analyze the sediment sample by using a sieve shaker. PDS 2000 software was used for data acquisition, and Qimera QPS version 2.4.5 was used for processing the bathymetry data. Meanwhile, FMGT QPS version 7.10 processes the backscatter data. Then, backscatter data were analyzed by using the maximum likelihood classification tool in ArcGIS version 10.8 software. The result identified three types of sediments around the coral which were very coarse sand, coarse sand, and medium sand.

Keywords: sediment type, MBES echo sounder, backscatter, ArcGIS

Procedia PDF Downloads 80
18 Learning Management System Technologies for Teaching Computer Science at a Distance Education Institution

Authors: Leila Goosen, Dalize van Heerden

Abstract:

The performance outcomes of first year Computer Science and Information Technology students across the world are of great concern, whether they are being taught in a face-to-face environment or via distance education. In the face-to-face environment, it is, however, somewhat easier to teach and support students than it is in a distance education environment. The face-to-face academic can more easily gauge the level of understanding and participation of students and implement interventions to address issues, which may arise. With the inroads that Web 2.0 and Web 3.0 technologies are making, the world of online teaching and learning are rapidly expanding, bringing about technologies, which allows for similar interactions between online academics and their students as available to their face-to-face counter parts. At the University of South Africa (UNISA), the Learning Management System (LMS) is called myUNISA and it is deployed on a SAKAI platform. In this paper, we will take a look at some of the myUNISA technologies implemented in the teaching of a first year programming course, how they are implemented and, in some cases, we will indicate how this affects the performance outcomes of students.

Keywords: computer science, Distance Education Technologies, Learning Management System, face-to-face environment

Procedia PDF Downloads 488
17 Formulating Rough Approximations in Information Tables with Possibilistic Information

Authors: Michinori Nakata, Hiroshi Sakai

Abstract:

A rough set, which consists of lower and upper approximations, is formulated in information tables containing possibilistic information. First, lower and upper approximations on the basis of possible world semantics in the same way as Lipski did in the field of incomplete databases are shown in order to clarify fundamentals of rough sets under possibilistic information. Possibility and necessity measures are used, as is done in possibilistic databases. As a result, each object has certain and possible membership degrees to lower and upper approximations, which degrees are the lower and upper bounds. Therefore, the degree that the object belongs to lower and upper approximations is expressed by an interval value. And the complementary property linked with the lower and upper approximations holds, as is valid under complete information. Second, the approach based on indiscernibility relations, which is proposed by Dubois and Prade, are extended in three cases. The first case is that objects used to approximate a set of objects are characterized by possibilistic information. The second case is that objects used to approximate a set of objects with possibilistic information are characterized by complete information. The third case is that objects that are characterized by possibilistic information approximate a set of objects with possibilistic information. The extended approach create the same results as the approach based on possible world semantics. This justifies our extension.

Keywords: rough sets, possibilistic information, possible world semantics, indiscernibility relations, lower approximations, upper approximations

Procedia PDF Downloads 318
16 Control of a Plane Jet Spread by Tabs at the Nozzle Exit

Authors: Makito Sakai, Takahiro Kiwata, Takumi Awa, Hiroshi Teramoto, Takaaki Kono, Kuniaki Toyoda

Abstract:

Using experimental and numerical results, this paper describes the effects of tabs on the flow characteristics of a plane jet at comparatively low Reynolds numbers while focusing on the velocity field and the vortical structure. The flow visualization and velocity measurements were respectively carried out using laser Doppler velocimetry (LDV) and particle image velocimetry (PIV). In addition, three-dimensional (3D) plane jet numerical simulations were performed using ANSYS Fluent, a commercially available computational fluid dynamics (CFD) software application. We found that the spreads of jets perturbed by large delta tabs and round tabs were larger than those produced by the other tabs tested. Additionally, it was determined that a plane jet with square tabs had the smallest jet spread downstream, and the jet’s centerline velocity was larger than those of jets perturbed by the other tabs tested. It was also observed that the spanwise vortical structure of a plane jet with tabs disappeared completely. Good agreement was found between the experimental and numerical simulation velocity profiles in the area near the nozzle exit when the laminar flow model was used. However, we also found that large eddy simulation (LES) is better at predicting the developing flow field of a plane jet than the laminar and the standard k-ε turbulent models.

Keywords: plane jet, flow control, tab, flow measurement, numerical simulation

Procedia PDF Downloads 331
15 Morphology of the Acetabular Cartilage Surface in Elderly Cadavers Analyzing the Contact between the Acetabulum and Femoral Head

Authors: Keisuke Akiyama, Takashi Sakai, Junichiro Koyanagi, Hideki Yoshikawa, Kazuomi Sugamoto

Abstract:

The geometry of acetabular cartilage surface plays an important role in hip joint biomechanics. The aim of this study was to analyze the morphology of acetabular articular cartilage surface in elderly subjects using a 3D-digitizer. Twenty hemipelves from 12 subjects (mean ages 85 years) were scanned with 3D-digitizer. Each acetabular surface model was divided into four regions: anterosuperior (AS), anteroinferior (AI), posterosuperior (PS), and posteroinferior (PI). In the global acetabulum and each region, the acetabular sphere radius and the standard deviation (SD) of the distance from the acetabular sphere center to the acetabular cartilage surface were calculated. In the global acetabulum, the distance between the acetabular surface model and the maximum sphere which did not penetrate over the acetabular surface model was calculated as the inferred femoral head, and then the distribution was mapped at intervals of 0.5 mm. The SD in AS was significantly larger than that in AI (p = 0.006) and PI (p = 0.001). The SD in PS was significantly larger than that in PI (p = 0.005). The closest region (0-0.5 mm) tended to be distributed at anterior or posterosuperior acetabular edge. The contact between the femoral head and acetabulum might start at the periphery of the lunate surface, especially in the anterior or posterosuperior region. From viewpoint of acetabular morphology, the acetabular articular cartilage in the anterior or posterosuperior edge could be more vulnerable due to direct contact mechanism.

Keywords: acetabulum, cartilage, morphology, 3D-digitizer

Procedia PDF Downloads 340
14 Association between Severe Acidemia before Endotracheal Intubation and the Lower First Attempt Intubation Success Rate

Authors: Keiko Naito, Y. Nakashima, S. Yamauchi, Y. Kunitani, Y. Ishigami, K. Numata, M. Mizobe, Y. Homma, J. Takahashi, T. Inoue, T. Shiga, H. Funakoshi

Abstract:

Background: A presence of severe acidemia, defined as pH < 7.2, is common during endotracheal intubation for critically ill patients in the emergency department (ED). Severe acidemia is widely recognized as a predisposing factor for intubation failure. However, it is unclear that acidemic condition itself actually makes endotracheal intubation more difficult. We aimed to evaluate if a presence of severe acidemia before intubation is associated with the lower first attempt intubation success rate in the ED. Methods: This is a retrospective observational cohort study in the ED of an urban hospital in Japan. The collected data included patient demographics, such as age, sex, and body mass index, presence of one or more factors of modified LEMON criteria for predicting difficult intubation, reasons for intubation, blood gas levels, airway equipment, intubation by emergency physician or not, and the use of the rapid sequence intubation technique. Those with any of the following were excluded from the analysis: (1) no blood gas drawn before intubation, (2) cardiopulmonary arrest, and (3) under 18 years of age. The primary outcome was the first attempt intubation success rates between a severe acidemic patients (SA) group and a non-severe acidemic patients (NA) group. Logistic regression analysis was used to test the first attempt success rates for intubations between those two groups. Results: Over 5 years, a total of 486 intubations were performed; 105 in the SA group and 381 in the NA group. The univariate analysis showed that the first attempt intubation success rate was lower in the SA group than in the NA group (71.4% vs 83.5%, p < 0.01). The multivariate logistic regression analysis identified that severe acidemia was significantly associated with the first attempt intubation failure (OR 1.9, 95% CI 1.03-3.68, p = 0.04). Conclusions: A presence of severe acidemia before endotracheal intubation lowers the first attempt intubation success rate in the ED.

Keywords: acidemia, airway management, endotracheal intubation, first-attempt intubation success rate

Procedia PDF Downloads 242
13 A Challenge to Acquire Serious Victims’ Locations during Acute Period of Giant Disasters

Authors: Keiko Shimazu, Yasuhiro Maida, Tetsuya Sugata, Daisuke Tamakoshi, Kenji Makabe, Haruki Suzuki

Abstract:

In this paper, we report how to acquire serious victims’ locations in the Acute Stage of Large-scale Disasters, in an Emergency Information Network System designed by us. The background of our concept is based on the Great East Japan Earthquake occurred on March 11th, 2011. Through many experiences of national crises caused by earthquakes and tsunamis, we have established advanced communication systems and advanced disaster medical response systems. However, Japan was devastated by huge tsunamis swept a vast area of Tohoku causing a complete breakdown of all the infrastructures including telecommunications. Therefore, we noticed that we need interdisciplinary collaboration between science of disaster medicine, regional administrative sociology, satellite communication technology and systems engineering experts. Communication of emergency information was limited causing a serious delay in the initial rescue and medical operation. For the emergency rescue and medical operations, the most important thing is to identify the number of casualties, their locations and status and to dispatch doctors and rescue workers from multiple organizations. In the case of the Tohoku earthquake, the dispatching mechanism and/or decision support system did not exist to allocate the appropriate number of doctors and locate disaster victims. Even though the doctors and rescue workers from multiple government organizations have their own dedicated communication system, the systems are not interoperable.

Keywords: crisis management, disaster mitigation, messing, MGRS, military grid reference system, satellite communication system

Procedia PDF Downloads 234
12 Thoughts Regarding Interprofessional Work between Nurses and Speech-Language-Hearing Therapists in Cancer Rehabilitation: An Approach for Dysphagia

Authors: Akemi Nasu, Keiko Matsumoto

Abstract:

Rehabilitation for cancer requires setting up individual goals for each patient and an approach that properly fits the stage of cancer when putting into practice. In order to cope with the daily changes in the patients' condition, the establishment of a good cooperative relationship between the nurses and the physiotherapists, occupational therapists, and speech-language-hearing therapists (therapists) becomes essential. This study will focus on the present situation of the cooperation between nurses and therapists, especially the speech-language-hearing therapists, and aim to elucidate what develops there. A semi-structured interview was conducted targeted at a physical therapist having practical experience in working in collaboration with nurses. The contents of the interview were transcribed and converted to data, and the data was encoded and categorized with sequentially increasing degrees of abstraction to conduct a qualitative explorative factor analysis of the data. When providing ethical explanations, particular care was taken to ensure that participants would not be subjected to any disadvantages as a result of participating in the study. In addition, they were also informed that their privacy would be ensured and that they have the right to decline to participate in the study. In addition, they were also informed that the results of the study would be announced publicly at an applicable nursing academic conference. This study has been approved following application to the ethical committee of the university with which the researchers are affiliated. The survey participant is a female speech-language-hearing therapist in her forties. As a result of the analysis, 6 categories were extracted consisting of 'measures to address appetite and aspiration pneumonia prevention', 'limitation of the care a therapist alone could provide', 'the all-inclusive patient- supportive care provided by nurses', 'expand the beneficial cooperation with nurses', 'providing education for nurses on the swallowing function utilizing videofluoroscopic examination of swallowing', 'enhancement of communication including conferences'. In order to improve the team performance, and for the teamwork competency necessary for the provision of safer care, mutual support is essential. As for the cooperation between nurses and therapists, this survey indicates that the maturing of the cooperation between professionals in order to improve nursing professionals' knowledge and enhance communication will lead to an improvement in the quality of the rehabilitation for cancer.

Keywords: cancer rehabilitation, nurses, speech-language-hearing therapists, interprofessional work

Procedia PDF Downloads 131
11 Postmortem Genetic Testing to Sudden and Unexpected Deaths Using the Next Generation Sequencing

Authors: Eriko Ochiai, Fumiko Satoh, Keiko Miyashita, Yu Kakimoto, Motoki Osawa

Abstract:

Sudden and unexpected deaths from unknown causes occur in infants and youths. Recently, molecular links between a part of these deaths and several genetic diseases are examined in the postmortem. For instance, hereditary long QT syndrome and Burgada syndrome are occasionally fatal through critical ventricular tachyarrhythmia. There are a large number of target genes responsible for such diseases, the conventional analysis using the Sanger’s method has been laborious. In this report, we attempted to analyze sudden deaths comprehensively using the next generation sequencing (NGS) technique. Multiplex PCR to subject’s DNA was performed using Ion AmpliSeq Library Kits 2.0 and Ion AmpliSeq Inherited Disease Panel (Life Technologies). After the library was constructed by emulsion PCR, the amplicons were sequenced 500 flows on Ion Personal Genome Machine System (Life Technologies) according to the manufacture instruction. SNPs and indels were analyzed to the sequence reads that were mapped on hg19 of reference sequences. This project has been approved by the ethical committee of Tokai University School of Medicine. As a representative case, the molecular analysis to a 40 years old male who received a diagnosis of Brugada syndrome demonstrated a total of 584 SNPs or indels. Non-synonymous and frameshift nucleotide substitutions were selected in the coding region of heart disease related genes of ANK2, AKAP9, CACNA1C, DSC2, KCNQ1, MYLK, SCN1B, and STARD3. In particular, c.629T-C transition in exon 3 of the SCN1B gene, resulting in a leu210-to-pro (L210P) substitution is predicted “damaging” by the SIFT program. Because the mutation has not been reported, it was unclear if the substitution was pathogenic. Sudden death that failed in determining the cause of death constitutes one of the most important unsolved subjects in forensic pathology. The Ion AmpliSeq Inherited Disease Panel can amplify the exons of 328 genes at one time. We realized the difficulty in selection of the true source from a number of candidates, but postmortem genetic testing using NGS analysis deserves of a diagnostic to date. We now extend this analysis to SIDS suspected subjects and young sudden death victims.

Keywords: postmortem genetic testing, sudden death, SIDS, next generation sequencing

Procedia PDF Downloads 354
10 Differences in Patient Satisfaction Observed between Female Japanese Breast Cancer Patients Who Receive Breast-Conserving Surgery or Total Mastectomy

Authors: Keiko Yamauchi, Motoyuki Nakao, Yoko Ishihara

Abstract:

The increase in the number of women with breast cancer in Japan has required hospitals to provide a higher quality of medicine so that patients are satisfied with the treatment they receive. However, patients’ satisfaction following breast cancer treatment has not been sufficiently studied. Hence, we investigated the factors influencing patient satisfaction following breast cancer treatment among Japanese women. These women underwent either breast-conserving surgery (BCS) (n = 380) or total mastectomy (TM) (n = 247). In March 2016, we conducted a cross-sectional internet survey of Japanese women with breast cancer in Japan. We assessed the following factors: socioeconomic status, cancer-related information, the role of medical decision-making, the degree of satisfaction regarding the treatments received, and the regret arising from the medical decision-making processes. We performed logistic regression analyses with the following dependent variables: extreme satisfaction with the treatments received, and regret regarding the medical decision-making process. For both types of surgery, the odds ratio (OR) of being extremely satisfied with the cancer treatment was significantly higher among patients who did not have any regrets compared to patients who had. Also, the OR tended to be higher among patients who chose to play a wanted role in the medical decision-making process, compared with patients who did not. In the BCS group, the OR of being extremely satisfied with the treatment was higher if, at diagnosis, the patient’s youngest child was older than 19 years, compared with patients with no children. The OR was also higher if patient considered the stage and characteristics of their cancer significant. The OR of being extremely satisfied with the treatments was lower among patients who were not employed on full-time basis, and among patients who considered the second medical opinions and medical expenses to be significant. These associations were not observed in the TM group. The OR of having regrets regarding the medical decision-making process was higher among patients who chose to play a role in the decision-making process as they preferred, and was also higher in patients who were employed on either a part-time or contractual basis. For both types of surgery, the OR was higher among patients who considered a second medical opinion to be significant. Regardless of surgical type, regret regarding the medical decision-making process decreases treatment satisfaction. Patients who received breast-conserving surgery were more likely to have regrets concerning the medical decision-making process if they could not play a role in the process as they preferred. In addition, factors associated with the satisfaction with treatment in BCS group but not TM group included the second medical opinion, medical expenses, employment status, and age of the youngest child at diagnosis.

Keywords: medical decision making, breast-conserving surgery, total mastectomy, Japanese

Procedia PDF Downloads 144
9 Developing Methodology of Constructing the Unified Action Plan for External and Internal Risks in University

Authors: Keiko Tamura, Munenari Inoguchi, Michiyo Tsuji

Abstract:

When disasters occur, in order to raise the speed of each decision making and response, it is common that delegation of authority is carried out. This tendency is particularly evident when the department or branch of the organization are separated by the physical distance from the main body; however, there are some issues to think about. If the department or branch is too dependent on the head office in the usual condition, they might feel lost in the disaster response operation when they are face to the situation. Avoiding this problem, an organization should decide how to delegate the authority and also who accept the responsibility for what before the disaster. This paper will discuss about the method which presents an approach for executing the delegation of authority process, implementing authorities, management by objectives, and preparedness plans and agreement. The paper will introduce the examples of efforts for the three research centers of Niigata University, Japan to arrange organizations capable of taking necessary actions for disaster response. Each center has a quality all its own. One is the center for carrying out the research in order to conserve the crested ibis (or Toki birds in Japanese), the endangered species. The another is the marine biological laboratory. The third one is very unique because of the old growth forests maintained as the experimental field. Those research centers are in the Sado Island, located off the coast of Niigata Prefecture, is Japan's second largest island after Okinawa and is known for possessing a rich history and culture. It takes 65 minutes jetfoil (high-speed ferry) ride to get to Sado Island from the mainland. The three centers are expected to be easily isolated at the time of a disaster. A sense of urgency encourages 3 centers in the process of organizational restructuring for enhancing resilience. The research team from the risk management headquarters offer those procedures; Step 1: Offer the hazard scenario based on the scientific evidence, Step 2: Design a risk management organization for disaster response function, Step 3: Conduct the participatory approach to make consensus about the overarching objectives, Step 4: Construct the unified operational action plan for 3 centers, Step 5: Simulate how to respond in each phase based on the understanding the various phases of the timeline of a disaster. Step 6: Document results to measure performance and facilitate corrective action. This paper shows the result of verifying the output and effects.

Keywords: delegation of authority, disaster response, risk management, unified command

Procedia PDF Downloads 120
8 Evaluation Method for Fouling Risk Using Quartz Crystal Microbalance

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

Procedia PDF Downloads 208
7 Suggestion of Methodology to Detect Building Damage Level Collectively with Flood Depth Utilizing Geographic Information System at Flood Disaster in Japan

Authors: Munenari Inoguchi, Keiko Tamura

Abstract:

In Japan, we were suffered by earthquake, typhoon, and flood disaster in 2019. Especially, 38 of 47 prefectures were affected by typhoon #1919 occurred in October 2019. By this disaster, 99 people were dead, three people were missing, and 484 people were injured as human damage. Furthermore, 3,081 buildings were totally collapsed, 24,998 buildings were half-collapsed. Once disaster occurs, local responders have to inspect damage level of each building by themselves in order to certificate building damage for survivors for starting their life reconstruction process. At that disaster, the total number to be inspected was so high. Based on this situation, Cabinet Office of Japan approved the way to detect building damage level efficiently, that is collectively detection. However, they proposed a just guideline, and local responders had to establish the concrete and infallible method by themselves. Against this issue, we decided to establish the effective and efficient methodology to detect building damage level collectively with flood depth. Besides, we thought that the flood depth was relied on the land height, and we decided to utilize GIS (Geographic Information System) for analyzing the elevation spatially. We focused on the analyzing tool of spatial interpolation, which is utilized to survey the ground water level usually. In establishing the methodology, we considered 4 key-points: 1) how to satisfy the condition defined in the guideline approved by Cabinet Office for detecting building damage level, 2) how to satisfy survivors for the result of building damage level, 3) how to keep equitability and fairness because the detection of building damage level was executed by public institution, 4) how to reduce cost of time and human-resource because they do not have enough time and human-resource for disaster response. Then, we proposed a methodology for detecting building damage level collectively with flood depth utilizing GIS with five steps. First is to obtain the boundary of flooded area. Second is to collect the actual flood depth as sampling over flooded area. Third is to execute spatial analysis of interpolation with sampled flood depth to detect two-dimensional flood depth extent. Fourth is to divide to blocks by four categories of flood depth (non-flooded, over the floor to 100 cm, 100 cm to 180 cm and over 180 cm) following lines of roads for getting satisfaction from survivors. Fifth is to put flood depth level to each building. In Koriyama city of Fukushima prefecture, we proposed the methodology of collectively detection for building damage level as described above, and local responders decided to adopt our methodology at typhoon #1919 in 2019. Then, we and local responders detect building damage level collectively to over 1,000 buildings. We have received good feedback that the methodology was so simple, and it reduced cost of time and human-resources.

Keywords: building damage inspection, flood, geographic information system, spatial interpolation

Procedia PDF Downloads 120
6 UKIYO-E: User Knowledge Improvement Based on Youth Oriented Entertainment, Art Appreciation Support by Interacting with Picture

Authors: Haruya Tamaki, Tsugunosuke Sakai, Ryuichi Yoshida, Ryohei Egusa, Shigenori Inagaki, Etsuji Yamaguchi, Fusako Kusunoki, Miki Namatame, Masanori Sugimoto, Hiroshi Mizoguchi

Abstract:

Art appreciation is important as part of children education. Art appreciation can enrich sensibility and creativity. To enrich sensibility and creativity, the children have to learning knowledge of picture such as social and historical backgrounds and author intention. High learning effect can acquire by actively learning. In short, it is important that encourage learning of the knowledge about pictures actively. It is necessary that children feel like interest to encourage learning of the knowledge about pictures actively. In a general art museum, comments on pictures are done through writing. Thus, we expect that this method cannot arouse the interest of the children in pictures, because children feel like boring. In brief, learning about the picture information is difficult. Therefore, we are developing an art-appreciation support system that will encourage learning of the knowledge about pictures actively by children feel like interest. This system uses that Interacting with Pictures to learning of the knowledge about pictures. To Interacting with Pictures, children have to utterance by themselves. We expect that will encourage learning of the knowledge about pictures actively by Interacting with Pictures. To more actively learning, children can choose who talking with by information that location and movement of the children. This system must be able to acquire real-time knowledge of the location, movement, and voice of the children. We utilize the Microsoft’s Kinect v2 sensor and its library, namely, Kinect for Windows SDK and Speech Platform SDK v11 for this purpose. By using these sensor and library, we can determine the location, movement, and voice of the children. As the first step of this system, we developed ukiyo-e game that use ukiyo-e to appreciation object. Ukiyo-e is a traditional Japanese graphic art that has influenced the western society. Therefore, we believe that the ukiyo-e game will be appreciated. In this study, we applied talking to pictures to learn information about the pictures because we believe that learning information about the pictures by talking to the pictures is more interesting than commenting on the pictures using only texts. However, we cannot confirm if talking to the pictures is more interesting than commenting using texts only. Thus, we evaluated through EDA measurement whether the user develops an interest in the pictures while talking to them using voice recognition or by commenting on the pictures using texts only. Hence, we evaluated that children have interest to picture while talking to them using voice recognition through EDA measurement. In addition, we quantitatively evaluate that enjoyed this game or not and learning information about the pictures for primary schoolchildren. In this paper, we summarize these two evaluation results.

Keywords: actively learning, art appreciation, EDA, Kinect V2

Procedia PDF Downloads 280
5 Acid Soil Amelioration Using Coal Bio-Briquette Ash and Waste Concrete in China

Authors: Y. Sakai, C. Wang

Abstract:

The decrease in agricultural production due to soil deterioration has been an urgent task. Soil acidification is a potentially serious land degradation issue and it will have a major impact on agricultural productivity and sustainable farming systems. In China, acid soil is mainly distributed in the southern part, the decrease in agricultural production and heavy metal contamination are serious problems. In addition, not only environmental and health problems due to the exhaust gas such as mainly sulfur dioxide (SO₂) but also the generation of a huge amount of construction and demolition wastes with the accelerating urbanization has emerged as a social problem in China. Therefore, the need for the recycling and reuse of both desulfurization waste and waste concrete is very urgent and necessary. So we have investigated the effectiveness as acid soil amendments of both coal bio-briquette ash and waste concrete. In this paper, acid soil (AS1) in Nanjing (pH=6.0, EC=1.6dSm-1) and acid soil (AS2) in Guangzhou (pH=4.1, EC=0.2dSm-1) were investigated in soil amelioration test. Soil amendments were three coal bio-briquette ashes (BBA1, BBA2 and BBA3), the waste cement fine powders (CFP) ( < 200µm (particle diameter)), waste concrete particles (WCP) ( < 4.75mm ( < 0.6mm, 0.6-1.0mm, 1.0-2.0mm, 2.0-4.75mm)), and six mixtures with two coal bio-briquette ashes (BBA2 and BBA3), CFP, WCP( < 0.6mm) and WCP(2.0-4.75mm). In acid soil amelioration test, the three BBAs, CFP and various WCPs based on exchangeable calcium concentration were added to two acid soils. The application rates were from 0 wt% to 3.5 wt% in AS1 test and from 0 wt% to 6.0 wt% in AS2 test, respectively. Soil chemical properties (pH, EC, exchangeable and soluble ions (Na, Ca, Mg, K)) before and after mixing with soil amendments were measured. In addition, Al toxicity and the balance of salts (CaO, K₂O, MgO) in soil after amelioration was evaluated. The order of pH and exchangeable Ca concentration that is effective for acid soil amelioration was WCP(0.6mm) > CFP > WCP(2.0-4.25mm) > BB1 > BB2 > BB3. In all AS 1 and AS 2 amelioration tests using three BBAs, the pH and EC increased slightly with the increase of application rate and reached to the appropriate value range of both pH and EC in BBA1 only. Because BBA1 was higher value in pH and exchangeable Ca. After that, soil pH and EC with the increase in the application rate of BBA2, BBA3 and by using CFP, WC( < 0.6mm), WC(2.0-4.75mm) as soil amendment reached to each appropriate value range, respectively. In addition, the mixture amendments with BBA2, BBA3 CFP, WC( < 0.6mm), and WC(2.0-4.75mm) could ameliorate at a smaller amount of application rate in case of BBA only. And the exchangeable Al concentration decreased drastically with the increase in pH due to soil amelioration and was under the standard value. Lastly, the heavy metal (Cd, As, Se, Ni, Cr, Pb, Mo, B, Cu, Zn) contents in new soil amendments were under control standard values for agricultural use in China. Thus we could propose a new acid soil amelioration method using coal bio-briquette ash and waste concrete in China.

Keywords: acid soil, coal bio-briquette ash, soil amelioration, waste concrete

Procedia PDF Downloads 179
4 Correlation between Body Mass Index and Blood Sugar/Serum Lipid Levels in Fourth-Grade Boys in Japan

Authors: Kotomi Yamashita, Hiromi Kawasaki, Satoko Yamasaki, Susumu Fukita, Risako Sakai

Abstract:

Lifestyle-related diseases develop from the long-term accumulation of health consequences from a poor lifestyle. Thus, schoolchildren, who have not accumulated long-term lifestyle habits, are believed to be at a lower risk for lifestyle-related diseases. However, schoolchildren rarely receive blood tests unless they are under treatment for a serious disease; without such data on their blood, the impacts of their young lifestyle could not be known. Blood data from physical measurements can help in the implementation of more effective health education. Therefore, we examined the correlation between body mass index (BMI) and blood sugar/serum lipid (BS/SL) levels. From 2014 to 2016, we measured the blood data of fourth-grade students living in a city in Japan. The present study reported on the results of 281 fourth-grade boys only (80.3% of total). We analyzed their BS/SL levels by comparing the blood data against the criteria of the National Center for Child Health and Development in Japan. Next, we examined the correlation between BMI and BS/SL levels. IBM SPSS Statistics for Windows, Version 25 was used for analysis. A total of 69 boys (24.6%) were within the normal range for BMI (18.5–24), whereas 193 (71.5%) and 8 boys (2.8%) had lower and higher BMI, respectively. Regarding BS levels, 280 boys were within the normal range (70–90 mg/dl); 1 boy reported a higher value. All the boys were within the normal range for glycated Hemoglobin (HbA1c) (4.6–6.2%). Regarding SL levels, 271 boys were within the normal range (125–230 mg/dl) for total cholesterol (TC), whereas 5 boys (1.8%) had lower and 5 boys (1.8%) had higher levels. A total of 243 boys (92.7%) were within the normal range (36-138mg/dL) for triglycerides (TG), whereas 19 boys (7.3%) had lower and 19 boys (7.3%) had higher levels. Regarding high-density lipoprotein cholesterol (HDL-C), 276 boys (98.2%) were within the normal range (40-mg/dl), whereas 5 boys (1.8%) reported lower values. All but one boy (280, 99.6%) were within the normal range (-170 mg/dl) for low-density lipoprotein cholesterol (LDL-C); the exception (0.4%) had a higher level. BMI and BS didn’t show a correlation. BMI and HbA1c were moderately positively correlated (r = 0.139, p=0.019). We also observed moderate positive correlations between BMI and TG (r = 0.328, p < 0.01), TC (r=0.239, p< 0.01), LDL-C (r = 0.324, p < 0.01), respectively. BMI and HDL-C were low correlated (r = -0.185, p = 0.002). Most of the boys were within the normal range for BS/SL levels. However, some boys exceeded the normal TG range. Fourth graders with a high TG may develop a lifestyle-related disease in the future. Given its relation to TG, food habits should be improved in this group. Our findings suggested a positive correlation between BMI and BS/SL levels. Fourth-grade schoolboys with a high BMI may be at high risk for developing lifestyle-related diseases. Lifestyle improvement may be recommended to lower the BS/SL levels in this group.

Keywords: blood sugar level, lifestyle-related diseases, school students, serum lipid level

Procedia PDF Downloads 135
3 Application of Fatty Acid Salts for Antimicrobial Agents in Koji-Muro

Authors: Aya Tanaka, Mariko Era, Shiho Sakai, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

Abstract:

Objectives: Aspergillus niger and Aspergillus oryzae are used as koji fungi in the spot of the brewing. Since koji-muro (room for making koji) was a low level of airtightness, microbial contamination has long been a concern to the alcoholic beverage production. Therefore, we focused on the fatty acid salt which is the main component of soap. Fatty acid salts have been reported to show some antibacterial and antifungal activity. So this study examined antimicrobial activities against Aspergillus and Bacillus spp. This study aimed to find the effectiveness of the fatty acid salt in koji-muro as antimicrobial agents. Materials & Methods: A. niger NBRC 31628, A. oryzae NBRC 5238, A. oryzae (Akita Konno store) and Bacillus subtilis NBRC 3335 were chosen as tested. Nine fatty acid salts including potassium butyrate (C4K), caproate (C6K), caprylate (C8K), caprate (C10K), laurate (C12K), myristate (C14K), oleate (C18:1K), linoleate (C18:2K) and linolenate (C18:3K) at 350 mM and pH 10.5 were used as antimicrobial activity. FASs and spore suspension were prepared in plastic tubes. The spore suspension of each fungus (3.0×104 spores/mL) or the bacterial suspension (3.0×105 CFU/mL) was mixed with each of the fatty acid salts (final concentration of 175 mM). The mixtures were incubated at 25 ℃. Samples were counted at 0, 10, 60, and 180 min by plating (100 µL) on potato dextrose agar. Fungal and bacterial colonies were counted after incubation for 1 or 2 days at 30 ℃. The MIC (minimum inhibitory concentration) is defined as the lowest concentration of drug sufficient for inhibiting visible growth of spore after 10 min of incubation. MICs against fungi and bacteria were determined using the two-fold dilution method. Each fatty acid salt was separately inoculated with 400 µL of Aspergillus spp. or B. subtilis NBRC 3335 at 3.0 × 104 spores/mL or 3.0 × 105 CFU/mL. Results: No obvious change was observed in tested fatty acid salts against A. niger and A. oryzae. However, C12K was the antibacterial effect of 5 log-unit incubated time for 10 min against B. subtilis. Thus, C12K suppressed 99.999 % of bacterial growth. Besides, C10K was the antibacterial effect of 5 log-unit incubated time for 180 min against B. subtilis. C18:1K, C18:2K and C18:3K was the antibacterial effect of 5 log-unit incubated time for 10 min against B. subtilis. However, compared to saturated fatty acid salts to unsaturated fatty acid salts, saturated fatty acid salts are lower cost. These results suggest C12K has potential in the field of koji-muro. It is necessary to evaluate the antimicrobial activity against other fungi and bacteria, in the future.

Keywords: Aspergillus, antimicrobial, fatty acid salts, koji-muro

Procedia PDF Downloads 547
2 A High-Throughput Enzyme Screening Method Using Broadband Coherent Anti-stokes Raman Spectroscopy

Authors: Ruolan Zhang, Ryo Imai, Naoko Senda, Tomoyuki Sakai

Abstract:

Enzymes have attracted increasing attentions in industrial manufacturing for their applicability in catalyzing complex chemical reactions under mild conditions. Directed evolution has become a powerful approach to optimize enzymes and exploit their full potentials under the circumstance of insufficient structure-function knowledge. With the incorporation of cell-free synthetic biotechnology, rapid enzyme synthesis can be realized because no cloning procedure such as transfection is needed. Its open environment also enables direct enzyme measurement. These properties of cell-free biotechnology lead to excellent throughput of enzymes generation. However, the capabilities of current screening methods have limitations. Fluorescence-based assay needs applicable fluorescent label, and the reliability of acquired enzymatic activity is influenced by fluorescent label’s binding affinity and photostability. To acquire the natural activity of an enzyme, another method is to combine pre-screening step and high-performance liquid chromatography (HPLC) measurement. But its throughput is limited by necessary time investment. Hundreds of variants are selected from libraries, and their enzymatic activities are then identified one by one by HPLC. The turn-around-time is 30 minutes for one sample by HPLC, which limits the acquirable enzyme improvement within reasonable time. To achieve the real high-throughput enzyme screening, i.e., obtain reliable enzyme improvement within reasonable time, a widely applicable high-throughput measurement of enzymatic reactions is highly demanded. Here, a high-throughput screening method using broadband coherent anti-Stokes Raman spectroscopy (CARS) was proposed. CARS is one of coherent Raman spectroscopy, which can identify label-free chemical components specifically from their inherent molecular vibration. These characteristic vibrational signals are generated from different vibrational modes of chemical bonds. With the broadband CARS, chemicals in one sample can be identified from their signals in one broadband CARS spectrum. Moreover, it can magnify the signal levels to several orders of magnitude greater than spontaneous Raman systems, and therefore has the potential to evaluate chemical's concentration rapidly. As a demonstration of screening with CARS, alcohol dehydrogenase, which converts ethanol and nicotinamide adenine dinucleotide oxidized form (NAD+) to acetaldehyde and nicotinamide adenine dinucleotide reduced form (NADH), was used. The signal of NADH at 1660 cm⁻¹, which is generated from nicotinamide in NADH, was utilized to measure the concentration of it. The evaluation time for CARS signal of NADH was determined to be as short as 0.33 seconds while having a system sensitivity of 2.5 mM. The time course of alcohol dehydrogenase reaction was successfully measured from increasing signal intensity of NADH. This measurement result of CARS was consistent with the result of a conventional method, UV-Vis. CARS is expected to have application in high-throughput enzyme screening and realize more reliable enzyme improvement within reasonable time.

Keywords: Coherent Anti-Stokes Raman Spectroscopy, CARS, directed evolution, enzyme screening, Raman spectroscopy

Procedia PDF Downloads 135