Search results for: Satoko Fujiwara

Authors: Elsa Reethu, S. Karthik Babu, N. Syed

Abstract:

Background: Studies in the past focused on the additional benefits of action observation in improving upper and lower limb functions and improving activities of daily living when administered along with conventional therapy. Nevertheless, there is a paucity of literature proving the effects of motor imagery of truncal exercise in improving trunk control in patients with stroke. Aims/purpose: To study the effect of motor imagery of truncal exercises on trunk function and balance in early stroke. Methods: A total of 24 patients were included in the study. 12 were included in the experimental group and 12 were included in control group Trunk function was measured using Trunk Control Test (TCT), Trunk Impairment Scale Verheyden (TIS Verheyden) and Trunk Impairment Scale Fujiwara (TIS Fujiwara). The balance was assessed using Brunel Balance Assessment (BBA) and Tinetti POMA. For the experimental group, each session was for 30 minutes of physical exercises and 15 minutes of motor imagery, once a day, six times a week for 3 weeks and prior to the exercise session, patients viewed a video tape of all the trunk exercises to be performed for 15minutes. The control group practiced the trunk exercises alone for the same duration. Measurements were taken before, after and 4 weeks after intervention. Results: The effect of treatment in motor imagery group showed better improvement when compared with control group when measured after 3 weeks on values of static sitting balance, dynamic balance, total TIS (Verheyden) score, BBA, Tinetti balance and gait with a large effect size of 0.86, 1.99, 1.69, 1.06, 1.63 and 0.97 respectively. The moderate effect size was seen in values of TIS Fujiwara (0.58) and small effect size was seen on TCT (0.12) and TIS coordination component (0.13).at the end of 4 weeks after intervention, the large effect size was identified on values of dynamic balance (2.06), total TIS score (1.59) and Tinetti balance (1.24). The moderate effect size was observed on BBA (0.62) and Tinetti gait (0.72). Conclusion: Trunk motor imagery is effective in improving trunk function and balance in patients with stroke and has a carryover effect in the aspects of mobility. The therapy gain that was observed during the time of discharge was seen to be maintained at the follow-up levels.

Keywords: stroke, trunk rehabilitation, trunk function, balance, motor imagery

Procedia PDF Downloads 286

Authors: Satoko Fujiwara, Kiyotaka Obunai, Kazuya Okubo

Abstract:

A skew is one of important problems for designing the conveyor and transmission with frictional flat belt, in which running belt is deviated in width direction due to the transverse force applied to the belt. The skew often not only degrades the stability of the path of belt but also causes some damages of the belt and auxiliary machines. However, the transverse behavior such as the skew has not been discussed quantitatively in detail for frictional belts. The objective of this study is to clarify the transverse behavior of frictional flat belt driven by tapered pulley. Commercially available rubber flat belt reinforced by polyamide film was prepared as the test belt where the thickness and length were 1.25 mm and 630 mm, respectively. Test belt was driven between two pulleys made of aluminum alloy, where diameter and inter-axial length were 50 mm and 150 mm, respectively. Some tapered pulleys were applied where tapered angles were 0 deg (for comparison), 2 deg, 4 deg, and 6 deg. In order to alternatively investigate the transverse behavior, the transverse force applied to the belt was measured when the skew was constrained at the string under driving state. The transverse force was measured by a load cell having free rollers contacting on the side surface of the belt when the displacement in the belt width direction was constrained. The conditions of observed bending stiffness in-plane of the belt were changed by preparing three types of belts (the width of the belt was 20, 30, and 40 mm) where their observed stiffnesses were changed. The contributions of the bending stiffness in-plane of belt and initial inter-axial force to the transverse were discussed in experiments. The inter-axial force was also changed by setting a distance (about 240 mm) between the two pulleys. Influence of observed bending stiffness in-plane of the belt and initial inter-axial force on the transverse force were investigated. The experimental results showed that the transverse force was increased with an increase of observed bending stiffness in-plane of the belt and initial inter-axial force. The transverse force acting on the belt running on the tapered pulley was classified into multiple components. Those were components of forces applied with the deflection of the inter-axial force according to the change of taper angle, the resultant force by the bending moment applied on the belt winding around the tapered pulley, and the reaction force applied due to the shearing deformation. The calculation result of the transverse force was almost agreed with experimental data when those components were formulated. It was also shown that the most contribution was specified to be the shearing deformation, regardless of the test conditions. This study found that transverse behavior of frictional flat belt driven by tapered pulley was explained by the summation of those components of forces.

Keywords: skew, frictional flat belt, transverse force, tapered pulley

Procedia PDF Downloads 135

Authors: Shunsuke Fujiwara, Takashi Kaburagi, Kazuyuki Kobayashi, Kajiro Watanabe, Yosuke Kurihara

Abstract:

This paper describes a novel sensor device, a pressure pulse wave meter, which uses a bidirectional condenser microphone. The microphone work as a microphone as well as a sensor with high gain over a wide frequency range; they are also highly reliable and economical. Currently aging is becoming a serious social issue in Japan causing increased medical expenses in the country. Hence, it is important for elderly citizens to check health condition at home, and to care the health conditions through daily monitoring. Given this circumstances, we developed a novel pressure pulse wave meter based on a bidirectional condenser microphone. This novel pressure pulse wave meter device is used as a measuring instrument of health conditions.

Keywords: bidirectional microphone, pressure pulse wave meter, health condition, novel sensor device

Procedia PDF Downloads 530

Authors: Shogo Kato, Daisuke Okamoto, Satoko Tsuru, Yoshinori Iizuka, Ryoko Shimono

Abstract:

Healthcare safety has been perceived important. It is essential to prevent troubles in healthcare processes for healthcare safety. Trouble prevention is based on trouble prediction using accumulated knowledge on processes, troubles, and countermeasures. However, information on troubles has not been accumulated in hospitals in the appropriate structure, and it has not been utilized effectively to prevent troubles. In the previous study, though a detailed knowledge acquisition process for trouble prediction was proposed, the knowledgebase for countermeasures was not involved. In this paper, we aim to propose the structure of the knowledgebase for countermeasures in the knowledge acquisition process for trouble prediction in healthcare process. We first design the structure of countermeasures and propose the knowledge representation form on countermeasures. Then, we evaluate the validity of the proposal, by applying it into an actual hospital.

Keywords: trouble prevention, knowledge structure, structured knowledge, reusable knowledge

Procedia PDF Downloads 350

Authors: Nathaniel C. Villanueva, Ian K. H. Chun, Alyssa S. Fujiwara, Emily R. Leibovitch, Brennan E. Yamamoto, Loren G. Yamamoto

Abstract:

Introduction: In previous studies of high school sports, over 15% of concussions were attributed to contact with the playing surface. While artificial turf fields are increasing in popularity due to lower maintenance costs, artificial turf has been associated with more ankle and knee injuries, with inconclusive data on concussions. In this study, natural grass and artificial football fields were compared in terms of deceleration on fall impact. Methods: Accelerometers were placed on the forehead, apex of the head, and right ear of a Century Body Opponent Bag (BOB) manikin. A Riddell HITS football helmet was secured onto the head of the manikin over the accelerometers. This manikin was dropped onto natural grass (n = 10) and artificial turf (n = 9) high school football fields. The manikin was dropped from a stationary position at a height of 60 cm onto its front, back, and left side. Each of these drops was conducted 10 times at the 40-yard line, 20-yard line, and endzone. The net deceleration on impact was calculated as a net vector from each of the three accelerometers’ x, y, and z vectors from the three different locations on the manikin’s head (9 vector measurements per drop). Results: Mean values for the multiple drops were calculated for each accelerometer and drop type for each field. All accelerometers in forward and backward falls and one accelerometer in side falls showed significantly greater impact force on synthetic turf compared to the natural grass surfaces. Conclusion: Impact force was higher on synthetic fields for all drop types for at least one of the accelerometer locations. These findings suggest that concussion risk might be higher for athletes playing on artificial turf fields.

Keywords: concussion, football, biomechanics, sports

Procedia PDF Downloads 137

Authors: Yukiko Fujiwara

Abstract:

As a language learning methodology, Content and Language Integrated Learning (CLIL) has become increasingly prevalent in Japan. Most CLIL classroom practice and its research are conducted in EFL fields. However, much less research has been done in the Japanese language learning setting. Therefore, there are still many issues to work out using CLIL in the Japanese language teaching (JLT) setting. it is expected that more research will be conducted on both authentically and academically. Under such circumstances, this is one of the few classroom-based CLIL researches experiments in JLT and aims to find an effective course design for a class with students at different proficiency levels. The class was called ‘Japanese culture A’. This class was offered as one of the elective classes for International exchange students at a Japanese university. The Japanese proficiency level of the class was above the Japanese Language Proficiency Test Level N3. Since the CLIL approach places importance on ‘authenticity’, the class was designed with materials and activities; such as books, magazines, a film and TV show and a field trip to Kyoto. On the field trip, students experienced making traditional Japanese desserts, by receiving guidance directly from a Japanese artisan. Through the course, designated task sheets were used so the teacher could get feedback from each student to grasp what the class proficiency gap was. After reading an article on Japanese culture, students were asked to write down the words they did not understand and what they thought they needed to learn. It helped both students and teachers to set learning goals and work together for it. Using questionnaires and interviews with students, this research examined whether the attempt was effective or not. Essays they wrote in class were also analyzed. The results from the students were positive. They were motivated by learning authentic, natural Japanese, and they thrived setting their own personal goals. Some students were motivated to learn Japanese by studying the language and others were motivated by studying the cultural context. Most of them said they learned better this way; by setting their own Japanese language and culture goals. These results will provide teachers with new insight towards designing class materials and activities that support students in a multilevel CLIL class.

Keywords: authenticity, CLIL, Japanese language and culture, multilevel class

Procedia PDF Downloads 231

Authors: Houda Mnif Sellami, Toshi Umehara, Yuriko Yamazaki, Reie Matoba, Anna Sakashita, Yoshimi Abe, Hiroyuki Otake, Satoko Morita, Yoshitaka Akiyama, Chieko Morisawa, Eiji Omura, Masako Yazawa, Yoshie Koike, Mitsugu Tokunaga, Seiki Wada, Shinya Minagawa, Masafumi Matsuda

Abstract:

Childhood obesity has been, for decades, a very serious public health problem worldwide. Some Asian countries have already reached alarming rates, as lifestyle changed dramatically in this part of the world. In many concerned countries, strategies including educational, promotional and awareness-raising activities have been established to combat obesity within kids. Objective: To estimate the obesity and also the underweight trends of Japanese kids from 5 to 17 years, by single year of age and by gender, over the last decade. Methods We used the data from the cross-sectional annual Nationwide surveys (National Nutrition Survey, Japan, Ministry of education, culture, sports, science and technology) conducted from 2007 to 2016. We compared trajectories of obesity prevalence, with the data on sex and age groups. We also analyzed energy and macronutrients intakes of Japanese kids using Ministry of Health, Labor and Welfare-Japan annual data, from 2007 to 2014. Results: From 2007 to 2016, Boys obesity was higher than Girls obesity for the over 6 YO participants. Both Boys and Girls obesity trends had 2 peaks of prevalence at (11-13 YO) and then at (15-16 YO). From 2007 to 2012, Kids obesity decreased considerably in both sex and all year of age; then obesity decline was more modest till 2016.On the other side, Kids underweight prevalence increased in both sexes. The macronutrients analyze couldn’t show an evident association between obesity trends and foods intake. Conclusion: Japanese kids’ obesity has been decreased since 2007, in opposition to some other countries reports. We didn’t find an observed association with food intake using Health Ministry data; we need further investigation to estimate energy intake, lifestyle and physical activity by year of age to know whether there is any possible relation.

Keywords: childhood, Japan, obesity, underweight

Procedia PDF Downloads 259

Authors: Susumu Fukita, Hiromi Kawasaki, Satoko Yamasaki, Kotomi Yamashita, Tomoko Iki

Abstract:

The increase in the number of workers with mental health problems has become an issue. Many workers work in small- and medium-sized enterprises, which often support local employment and economy, especially in underpopulated mountainous areas. It is important for managers to take mental health measures for employees since there is no budget to hire health staff in small- and medium-sized enterprises. It is necessary to understand the manager's attitude toward the mental health of employees and to publicly support the manager in promoting mental health measures for employees. The purpose of this study was to study the awareness of managers of small- and medium-sized enterprises regarding the mental health of employees and to consider support for managers to take measures for the mental health of employees. Semi-structured interviews were conducted with six managers of small- and medium-sized enterprises in underpopulated mountainous areas in November 2019. Managers were asked about their awareness of the mental health of their employees. Qualitative descriptive analysis was used, and subcategories and categories were extracted. Four categories emerged. Regarding the mental health of employees, the managers acknowledged that if the appearance and behavior of the employees do not interfere with their lives, the manager judges that the employees’ mental health is normal. It was also found that the managers acknowledged that there is a comfortable working environment due to the characteristics of the underpopulated mountainous area. On the other hand, the managers acknowledged that employees are dissatisfied with salaries and management systems. In addition, it was found the manager acknowledged that some employees retire due to mental health problems. Although managers recognized that employees may be dissatisfied with salaries, they also recognized that there was a comfortable working environment due to the characteristics of the areas, with good interpersonal relationships. Economic challenges are difficult to solve in underpopulated mountainous areas. It is useful to consider measures that take advantage of the characteristics of the areas where it is easy to work because of good relations with each other, for example, to create a family-like workplace culture where managers and employees can engage in daily conversation. The managers judged that the employees were in good health if there was no interference with their lives. However, it is too late to take measures at the stage when it becomes an obstacle to life. Therefore, it is necessary to provide training for managers to learn observation techniques by which they quickly notice changes in the situation of employees and give appropriate responses; and to set up a contact point for managers to consult. Local governments should actively provide public support such as training for managers and establishing consultation desks to maintain valuable employment and local economics in underpopulated mountainous areas.

Keywords: employer, mental health, small- and medium- sized enterprises, underpopulated areas

Procedia PDF Downloads 121

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 122

Authors: Junwen Chen, Celina Feleppa, Tingyue Sun, Satoko Sasagawa, Michael Smithson

Abstract:

School refusal behaviours refer to behaviours to avoid school attendance, chronic lateness in arriving at school, or regular early dismissal. Poor attendance in schools is highly correlated with anxiety, depression, suicide attempts, delinquency, violence, and substance use and abuse. Poor attendance is also a strong indicator of lower achievement in school, as well as problematic social-emotional development. Long-term consequences of school refusal behaviours include fewer opportunities for higher education, employment, and social difficulties, and high risks of later psychiatric illness. Given its negative impacts on youth educational outcomes and well-being, a thorough understanding of factors that are involved in the development of this phenomenon is warranted for developing effective management approaches. This study investigated parental and adolescent factors that may contribute to school refusal behaviours by specifically focusing on the role of parental and adolescents’ anxiety and depression, emotion dysregulation, and parental rearing style. Findings are expected to inform the identification of both parental and adolescents’ factors that may contribute to school refusal behaviours. This knowledge will enable novel and effective approaches that incorporate these factors to managing school refusal behaviours in adolescents, which in turn improve their school and daily functioning. Results are important for an integrative understanding of school refusal behaviours. Furthermore, findings will also provide information for policymakers to weigh the benefits of interventions targeting school refusal behaviours in adolescents. One-hundred-and-six adolescents aged 12-18 years (mean age = 14.79 years old, SD = 1.78, males = 44) and their parents (mean age = 47.49 years old, SD = 5.61, males = 27) completed an online questionnaire measuring both parental and adolescents’ anxiety, depression, emotion dysregulation, parental rearing styles, and adolescents’ school refusal behaviours. Adolescents with school refusal behaviours reported greater anxiety and depression, with their parents showing greater emotion dysregulation. Parental emotion dysregulation and adolescents’ anxiety and depression predicted school refusal behaviours independently. To date, only limited studies have investigated the interplay between parental and youth factors in relation to youth school refusal behaviours. Although parental emotion dysregulation has been investigated in relation to youth emotion dysregulation, little is known about its role in the context of school refusal. This study is one of the very few that investigated both parental and adolescent factors in relation to school refusal behaviours in adolescents. The findings support the theoretical models that emphasise the role of youth and parental psychopathology in school refusal behaviours. Future management of school refusal behaviours should target adolescents’ anxiety and depression while incorporating training for parental emotion regulation skills.

Keywords: adolescents, school refusal behaviors, parental factors, anxiety and depression, emotion dysregulation

Procedia PDF Downloads 100

Authors: Satoko Yamasaki, Hiromi Kawasaki, Kotomi Yamashita, Susumu Fukita, Kei Sounai

Abstract:

People are susceptible to rumors when the cause of their health problems is unknown or invisible. In order for individuals to be unaffected by rumors, they need basic knowledge and correct information. Community health nursing classes use cases where basic knowledge of radiation can be utilized on a regular basis, thereby teaching that basic knowledge is important in preventing anxiety caused by rumors. Nursing students need to learn that preventive activities are essential for public health nursing care. This is the same methodology used to reduce COVID-19 anxiety among individuals. This study verifies the learning effect concerning the basic knowledge of radiation necessary for case consultation by emergency distance learning. Sixty third-year nursing college students agreed to participate in this research. The knowledge tests conducted before and after classes were compared, with the chi-square test used for testing. There were five knowledge questions regarding distance lessons. This was considered to be 5% significant. The students’ reports which describe the results of responding to health consultations, were analyzed qualitatively and descriptively. In this case study, a person living in an area not affected by radiation was anxious about drinking water and, thus, consulted with a student. The contents of the lecture were selected the minimum amount of knowledge used for the answers of the consultant; specifically hot spots, internal exposure risk, food safety, characteristics of cesium-137, and precautions for counselors. Before taking the class, the most correctly answered question by students concerned daily behavior at risk of internal exposure (52.2%). The question with the fewest correct answers was the selection of places that are likely to be hot spots (3.4%). All responses increased significantly after taking the class (p < 0.001). The answers to the counselors, as written by the students, were 'Cesium is strongly bound to the soil, so it is difficult to transfer to water' and 'Water quality test results of tap water are posted on the city's website.' These were concrete answers obtained by using specialized knowledge. Even in emergency distance learning, the students gained basic knowledge regarding radiation and created a document to utilize said knowledge while assuming the situation concretely. It was thought that the flipped classroom method, even if conducted remotely, could maintain students' learning. It was thought that setting specific knowledge and scenes to be used would enhance the learning effect. By changing the case to concern that of the anxiety caused by infectious diseases, students may be able to effectively gain the basic knowledge to decrease the anxiety of residents due to infectious diseases.

Keywords: effect of class, emergency distance learning, nursing student, radiation

Procedia PDF Downloads 99

Authors: Hiromi Kawasaki, Satoko Yamasaki, Mika Iwasa, Tomoko Iki, Akiko Takaki

Abstract:

In basic nursing, the conditions necessary for maintaining human health -temperature, humidity, illumination, distance from others, noise, moisture, meals, and excretion- were explained. Nursing students often think of these conditions in the context of a hospital room. In order to make students think of these conditions in terms of an environment necessary for maintaining health and preventing illness for residents, in the third year of community health nursing, students learned how to assess and improve the environment -particularly via the case of shelters in the event of a disaster. The importance of environmental management has increased in 2020 as a preventive measure against COVID-19 infection. We verified the effect of the lessons, which was decided to be conducted through distance learning. Sixty third-year nursing college students consented to participate in this study. Environmental standard knowledge for conducting environmental assessment was examined before and after class, and the percentage of correct answers was compared. The χ² test was used for the test, with a 5% significance level employed. Measures were evaluated via a report submitted by the students after class. Student descriptions were analyzed both qualitatively and descriptively with respect to expected health problems and suggestions for improvement. Students have already learned about the environment in terms of basic nursing in their second year. The correct answers for external environmental values concerning interpersonal distance, illumination, noise, and room temperature (p < 0.001) increased significantly after taking the class. Humidity was registered 83.3% before class and 93.3% after class (p = 0.077). Regarding the body, the percentage of students who answered correctly was 70% or more, both before and after the class. The students’ reports included overcrowding, high humidity/high temperature, and the number of toilets as health hazards. Health disorders to be prevented were heat stroke, infectious diseases, and economy class syndrome; improvement methods were recommended for hyperventilation, stretching, hydration, and waiting at home. After the public health nursing class, the students were able to not only propose environmental management of a hospital room but also had an understanding of the environment in terms of the lives of individuals, environmental assessment, and solutions to health problems. The response rate for basic items learned in the second year was already high before and after class, and interpersonal distance and ventilation were described by students. Students were able to use what they learned in basic nursing about the standards of the human mind and body. In the external environment, the memory of specific numerical values was ambiguous. The environment of the hospital room is controlled, and interest in numerical values may decrease. Nursing staff needs to maintain and improve human health as well as hospital rooms. With COVID-19, it was thought that students would continue to not only consider this point in reference to hospital rooms but also in regard to places where people gather. Even in distance learning, students were able to learn the important issues and lessons.

Keywords: environmental assessment, evacuation center, nursing education, nursing students

Procedia PDF Downloads 82

Authors: Srijata Mitra, Mobina Parveen, Pranab Roy, Narayan Chandra Chattopadhyay

Abstract:

Chlorinated hydrocarbon can be a major pollution problem in groundwater as well as soil. Many people interact with these chemicals on daily accidentally or by professionally in the laboratory. One of the most common sources for Chlorinated hydrocarbon contamination of soil and groundwater are industrial effluents. The wide use and discharge of Trichloroethylene (TCE), a volatile chlorohydrocarbon from chemical industry, led to major water pollution in rural areas. TCE is an mainly used as an industrial metal degreaser in industries. Biotransformation of TCE to the potent carcinogen vinyl chloride (VC) by consortia of anaerobic bacteria might have role for the above purpose. For these reasons, the aim of current study was to isolate and characterized the genes involved in TCE metabolism and also to investigate the in silico study of those genes. To our knowledge, only one aromatic dioxygenase system, the toluene dioxygenase in Pseudomonas putida F1 has been shown to be involved in TCE degradation. This is first instance where Bacillus cereus group being used in biodegradation of trichloroethylene. A novel bacterial strain 2479 was isolated from oil depot site at Rajbandh, Durgapur (West Bengal, India) by enrichment culture technique. It was identified based on polyphasic approach and ribotyping. The bacterium was gram positive, rod shaped, endospore forming and capable of degrading trichloroethylene as the sole carbon source. On the basis of phylogenetic data and Fatty Acid Methyl Ester Analysis, strain 2479 should be placed within the genus Bacillus and species cereus. However, the present isolate (strain 2479) is unique and sharply different from the usual Bacillus strains in its biodegrading nature. Fujiwara test was done to estimate that the strain 2479 could degrade TCE efficiently. The gene for TCE biodegradation was PCR amplified from genomic DNA of Bacillus cereus 2479 by using todC1 gene specific primers. The 600bp amplicon was cloned into expression vector pUC I8 in the E. coli host XL1-Blue and expressed under the control of lac promoter and nucleotide sequence was determined. The gene sequence was deposited at NCBI under the Accession no. GU183105. In Silico approach involved predicting the physico-chemical properties of deduced Tce1 protein by using ProtParam tool. The tce1 gene contained 342 bp long ORF encoding 114 amino acids with a predicted molecular weight 12.6 kDa and the theoretical pI value of the polypeptide was 5.17, molecular formula: C559H886N152O165S8, total number of atoms: 1770, aliphatic index: 101.93, instability index: 28.60, Grand Average of Hydropathicity (GRAVY): 0.152. Three differentially expressed proteins (97.1, 40 and 30 kDa) were directly involved in TCE biodegradation, found to react immunologically to the antibodies raised against TCE inducible proteins in Western blot analysis. The present study suggested that cloned gene product (TCE1) was capable of degrading TCE as verified chemically.

Keywords: cloning, Bacillus cereus, in silico analysis, TCE

Procedia PDF Downloads 378

Authors: Shuo Li, Yannick Coffinier, Chann Lagadec, Fabrizio Cleri, Katsuhiko Nishiguchi, Akira Fujiwara, Soo Hyeon Kim, Nicolas Clément

Abstract:

The need for single cell detection and analysis techniques has increased in the past decades because of the heterogeneity of individual living cells, which increases the complexity of the pathogenesis of malignant tumors. In the search for early cancer detection, high-precision medicine and therapy, the technologies most used today for sensitive detection of target analytes and monitoring the variation of these species are mainly including two types. One is based on the identification of molecular differences at the single-cell level, such as flow cytometry, fluorescence-activated cell sorting, next generation proteomics, lipidomic studies, another is based on capturing or detecting single tumor cells from fresh or fixed primary tumors and metastatic tissues, and rare circulating tumors cells (CTCs) from blood or bone marrow, for example, dielectrophoresis technique, microfluidic based microposts chip, electrochemical (EC) approach. Compared to other methods, EC sensors have the merits of easy operation, high sensitivity, and portability. However, despite various demonstrations of low limits of detection (LOD), including aptamer sensors, arrayed EC sensors for detecting single-cell have not been demonstrated. In this work, a new technique based on 20-nm-thick nanopillars array to support cells and keep them at ideal recognition distance for redox-labeled aptamers grafted on the surface. The key advantages of this technology are not only to suppress the false positive signal arising from the pressure exerted by all (including non-target) cells pushing on the aptamers by downward force but also to stabilize the aptamer at the ideal hairpin configuration thanks to a confinement effect. With the first implementation of this technique, a LOD of 13 cells (with5.4 μL of cell suspension) was estimated. In further, the nanosupported cell technology using redox-labeled aptasensors has been pushed forward and fully integrated into a single-cell electrochemical aptasensor array. To reach this goal, the LOD has been reduced by more than one order of magnitude by suppressing parasitic capacitive electrochemical signals by minimizing the sensor area and localizing the cells. Statistical analysis at the single-cell level is demonstrated for the recognition of cancer cells. The future of this technology is discussed, and the potential for scaling over millions of electrodes, thus pushing further integration at sub-cellular level, is highlighted. Despite several demonstrations of electrochemical devices with LOD of 1 cell/mL, the implementation of single-cell bioelectrochemical sensor arrays has remained elusive due to their challenging implementation at a large scale. Here, the introduced nanopillar array technology combined with redox-labeled aptamers targeting epithelial cell adhesion molecule (EpCAM) is perfectly suited for such implementation. Combining nanopillar arrays with microwells determined for single cell trapping directly on the sensor surface, single target cells are successfully detected and analyzed. This first implementation of a single-cell electrochemical aptasensor array based on Brownian-fluctuating redox species opens new opportunities for large-scale implementation and statistical analysis of early cancer diagnosis and cancer therapy in clinical settings.

Keywords: bioelectrochemistry, aptasensors, single-cell, nanopillars

Procedia PDF Downloads 89