Search results for: Alyssa S. Fujiwara
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 13

Search results for: Alyssa S. Fujiwara

13 Impact Force Difference on Natural Grass Versus Synthetic Turf Football Fields

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

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12 Effect of Motor Imagery of Truncal Exercises on Trunk Function and Balance in Early Stroke: A Randomized Controlled Trial

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

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11 Evaluation Using a Bidirectional Microphone as a Pressure Pulse Wave Meter

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

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10 Herbicide Resistant Weeds: Contrasting Perspectives of Actors in the Agricultural Sector

Authors: Bruce Small, Martin Espig, Alyssa Ryan

Abstract:

In the agricultural sector, the rapid expansion of herbicide resistant weeds is a major threat to the global sustainability of food and fibre production. Efforts to avoid herbicide resistance have primarily focused on new technologies and farmer education. Yet, despite decades of advice to growers from agricultural scientists and extension professionals of the need for management strategies for herbicide use, herbicide resistance continues to increase. Technological options are running out and current extension efforts to change farmer behaviour are failing to curb the problem. As part of a five-year, government funded, research programme to address herbicide resistance in New Zealand, social science theory and practice are being utilised to investigate the complexities of managing herbicide use and controlling resistance. As an initial step, we are utilising a transdisciplinary, multi-level systems approach to examine the problem definition, knowledge beliefs, attitudes and values of different important actors in the agri-business sector. In this paper, we report early project results from qualitative research examining the similarities and contrasts in the perceptions of scientists, farmer/growers, and rural professionals.

Keywords: behaviour change, herbicide resistant weeds, knowledge beliefs, systems perspective

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9 Teaching Method for a Classroom of Students at Different Language Proficiency Levels: Content and Language Integrated Learning in a Japanese Culture Classroom

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

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8 Learning and Teaching Styles of Student Nurses

Authors: Jefferson S. Galanza, Jewel An Mischelle R.Camcam, Alyssa Karryl C. Co, Stephanie P. De Guzman, Jet Jet K. Dongui-is, Rodolfo Dane C. Frias, Ovelle C. Jueco, Harvey L. Matbagan, Victoria Luzette T. Rillon, Christelle Romyna H. Saruca, Jeanette Roma M. Villasper

Abstract:

Background: Amidst numerous studies conducted on learning styles of students from a variety of courses, levels and school, a recent study recommended a great need for research on learning styles of student nurses. Moreover, related literatures have not been found exploring both the learning and teaching style of student nurses. Aims: The study aimed to determine the learning and teaching styles of student nurses and if there is an association between them. It also intended to discover whether student nurses are unimodal or multimodal in their styles and identified which faculty teaching style affords maximum outcome for student’s learning styles. Methods: Quantitative Descriptive-Correlational design was used. Participants were randomly selected 312 student nurses at School of Nursing X, Baguio City, Philippines. The questionnaire utilized a modified version of an adopted tool from Fleming’s VARK learning style version 7.2 (Visual, Auditory, Reader/Writer, Kinaesthetic) and Grasha’s teaching styles (Formal Authority, Demonstrator, Facilitator, Delegator). SPSS 19 was used for statistical treatment of data, where Chi square was used for the correlation of unimodal learning and teaching styles. Results/Finding: Majority of student nurses’ learning style is Kinesthetic and their teaching style is Demonstrator, which was also found to be significantly associated. Moreover, 8 out of 10 students are Unimodal in their learning and teaching modalities. In general, their preferred faculty teaching style is similar to their teaching style, which supports the concept, that teachers teach the way they learn. Conclusion: Study concludes that student nurses’ learning styles and teaching styles are varied, which exemplifies the uniqueness of every learner.This diversity in styles provided more evidence that a variety of mode of teaching and learning should be used by faculty and students to increase learning outcome and academic achievement. Recommendation: Future studies could be carried out in various schools of nursing utilizing faculty as respondents. Conduct assessment of learning style at the onset of classes/clinical placements so that faculty will become aware of the diversity of learners leading them to deliver diverse teaching methods.

Keywords: learning, learning styles, teaching styles, student nurses

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7 Acupuncture Reduces Pain Disability, Stress, and Depression in United States Military Veterans with Chronic Pain

Authors: Christine Eickhoff, Alyssa Adams, Alaine Duncan

Abstract:

The Washington, DC Veterans Affairs Medical Center (DC VAMC) offers complementary and integrative health (CIH) services such as acupuncture, yoga, meditation, and nutrition education through a coordinated outpatient clinic. The primary population utilizing CIH services are veterans with chronic pain. Acupuncture is one of the most popular of the CIH services available at the DC VAMC. As interest and availability grows, it is important to measure health outcomes associated with CIH service utilization. The purpose of this study was to investigate pain and mental health outcomes for veterans with chronic pain enrolled in individual acupuncture services in the DC VAMC. Veterans at the DC VAMC with self-identified chronic pain and no prior acupuncture experience were recruited for the study (n=70). Veterans were referred for services by a medical provider and completed baseline assessments at the program orientation prior to participating in any CIH services. Veterans received four individual, full-body acupuncture appointments within four weeks of study enrollment. After the first month, participants were scheduled for six appointments that occurred every two weeks and then eight more sessions that were scheduled one month apart. Follow-up assessments were administered at 2, 4, 6, 8, and 12 months. The findings reported will include completed time points at two and four months. Measures include a demographics survey, the Measure Yourself Medical Outcome Profile-2 (MYMOP-2), The Beck Depression Inventory (BDI-II), the Defense Veterans Pain Rating Scale (DVPRS), and the Pain Disability Questionnaire (PDQ). In this sample, 67% identified a pain condition as their primary health concern. Between baseline and two-month follow-up, there were significant improvements in participants’ primary health concern (MYMOP-2 p=0.010), general wellbeing (MYMOP-2 p=0.011), and a significant decrease in the use of medication (MYMOP-2 p<0.000). Between 2 and 4-month follow-up, pain disability (PDQ p=0.035), pain rating (DVPRS p=0.027), and depression (BDI-II p=0.003) significantly improved. Preliminary findings indicate that individual acupuncture therapy can be effective at improving health outcomes, well-being, and decreasing medication use in U.S. military veterans with chronic pain. Findings also suggest that individual acupuncture therapy can improve pain ratings, pain disability, and depression in veterans with chronic pain.

Keywords: acupuncture, chronic pain, depression, integrative health, medication use, military, pain, veterans, wellbeing

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6 Transverse Behavior of Frictional Flat Belt Driven by Tapered Pulley -Change of Transverse Force Under Driving State–

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

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5 Alcohol and Soda Consumption of University Students in Manila

Authors: Alexi Colleen F. Lim, Inna Felicia I. Agoncillo, Quenniejoy T. Dizon, Jennifer Joyce T. Eti, Carlota Aileen H. Monares, Neil Roy B. Rosales, Joshua F. Santillan, Alyssa Francesca D. S. Tanchuling, Josefina A. Tuazon, Mary Joan Therese C. Valera-Kourdache

Abstract:

Majority of leading causes of mortality in the Philippines are NCDs, which are preventable through control of known risk factors such as smoking, obesity, physical inactivity, and alcohol. Sugar-sweetened beverages such as soda and energy drinks also contribute to NCD risk and are of concern particularly for youth. This study provides baseline data on beverage consumption of university students in Manila with the focus on alcohol and soda. It further aims to identify factors affecting consumption. Specific objectives include: (1) to describe beverage consumption practices of university students in Manila; and (2) to determine factors promoting excessive consumption of alcohol and soda including demographic characteristics, attitude, interpersonal and environmental variables. Methods: The study employed correlational design with randomly selected students from two universities in Manila. Students 18 years or older who agreed to participate were included after obtaining ethical clearance. The study had two instruments: (1) World Health Organization’s Alcohol Use Disorders Identification Test (AUDIT) was used with permission, to determine excessive alcohol consumption; and (2) a questionnaire to obtain information regarding soda and energy drink consumption. Results: Out of 400 students surveyed, 70% were female and 78.75% were 18-20 years old (mean=19.79; SD=3.76). Among them, 51.50% consumed alcohol, with 30.10% excessive drinkers. Soda consumption is 91.50% with 37.70% excessive consumers. For energy drinks, 36.75% consume this and only 4.76% drink excessively. Using logistic regression, students who were more likely to be excessive alcohol drinkers belonged to non-health courses (OR=2.21) and purchased alcohol from bars (OR=7.84). Less likely to drink excessively are students who do not drink due to stress (OR=0.05) and drink when it is accessible (OR=0.02). Excessive soda consumption was less likely for female students (OR=0.28), those who drink when it is accessible (OR=0.14), do not drink soda during stressful situations (OR=0.19), and do not use soda as hangover treatment (OR=0.15). Conclusion: Excessive alcohol consumption was greater among students in Manila (30.10%) than in US (20%). Drinking alcohol with friends was not related to excessive consumption but availability in bars was. It is expected that health sciences students are less likely to engage in excessive alcohol as they are more aware of its ill effects. Prevalence of soda consumption in Manila (91.50%) is markedly higher compared to 24.5% in the US. These findings can inform schools in developing appropriate health education interventions and policies. For greater understanding of these behaviors and factors, further studies are recommended to explore knowledge and other factors that may promote excessive consumption.

Keywords: alcohol consumption, beverage consumption, energy drinks consumption, soda consumption, university students

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4 Laparoscopic Resection Shows Comparable Outcomes to Open Thoracotomy for Thoracoabdominal Neuroblastomas: A Meta-Analysis and Systematic Review

Authors: Peter J. Fusco, Dave M. Mathew, Chris Mathew, Kenneth H. Levy, Kathryn S. Varghese, Stephanie Salazar-Restrepo, Serena M. Mathew, Sofia Khaja, Eamon Vega, Mia Polizzi, Alyssa Mullane, Adham Ahmed

Abstract:

Background: Laparoscopic (LS) removal of neuroblastomas in children has been reported to offer favorable outcomes compared to the conventional open thoracotomy (OT) procedure. Critical perioperative measures such as blood loss, operative time, length of stay, and time to postoperative chemotherapy have all supported laparoscopic use rather than its more invasive counterpart. Herein, a pairwise meta-analysis was performed comparing perioperative outcomes between LS and OT in thoracoabdominal neuroblastoma cases. Methods: A comprehensive literature search was performed on PubMed, Ovid EMBASE, and Scopus databases to identify studies comparing the outcomes of pediatric patients with thoracoabdominal neuroblastomas undergoing resection via OT or LS. After deduplication, 4,227 studies were identified and subjected to initial title screening with exclusion and inclusion criteria to ensure relevance. When studies contained overlapping cohorts, only the larger series were included. Primary outcomes include estimated blood loss (EBL), hospital length of stay (LOS), and mortality, while secondary outcomes were tumor recurrence, post-operative complications, and operation length. The “meta” and “metafor” packages were used in R, version 4.0.2, to pool risk ratios (RR) or standardized mean differences (SMD) in addition to their 95% confidence intervals in the random effects model via the Mantel-Haenszel method. Heterogeneity between studies was assessed using the I² test, while publication bias was assessed via funnel plot. Results: The pooled analysis included 209 patients from 5 studies (141 OT, 68 LS). Of the included studies, 2 originated from the United States, 1 from Toronto, 1 from China, and 1was from a Japanese center. Mean age between study cohorts ranged from 2.4 to 5.3 years old, with female patients occupying between 30.8% to 50% of the study populations. No statistically significant difference was found between the two groups for LOS (SMD -1.02; p=0.083), mortality (RR 0.30; p=0.251), recurrence(RR 0.31; p=0.162), post-operative complications (RR 0.73; p=0.732), or operation length (SMD -0.07; p=0.648). Of note, LS appeared to be protective in the analysis for EBL, although it did not reach statistical significance (SMD -0.4174; p= 0.051). Conclusion: Despite promising literature assessing LS removal of pediatric neuroblastomas, results showed it was non-superior to OT for any explored perioperative outcomes. Given the limited comparative data on the subject, it is evident that randomized trials are necessary to further the efficacy of the conclusions reached.

Keywords: laparoscopy, neuroblastoma, thoracoabdominal, thoracotomy

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3 The Budget Impact of the DISCERN™ Diagnostic Test for Alzheimer’s Disease in the United States

Authors: Frederick Huie, Lauren Fusfeld, William Burchenal, Scott Howell, Alyssa McVey, Thomas F. Goss

Abstract:

Alzheimer’s Disease (AD) is a degenerative brain disease characterized by memory loss and cognitive decline that presents a substantial economic burden for patients and health insurers in the US. This study evaluates the payer budget impact of the DISCERN™ test in the diagnosis and management of patients with symptoms of dementia evaluated for AD. DISCERN™ comprises three assays that assess critical factors related to AD that regulate memory, formation of synaptic connections among neurons, and levels of amyloid plaques and neurofibrillary tangles in the brain and can provide a quicker, more accurate diagnosis than tests in the current diagnostic pathway (CDP). An Excel-based model with a three-year horizon was developed to assess the budget impact of DISCERN™ compared with CDP in a Medicare Advantage plan with 1M beneficiaries. Model parameters were identified through a literature review and were verified through consultation with clinicians experienced in diagnosis and management of AD. The model assesses direct medical costs/savings for patients based on the following categories: •Diagnosis: costs of diagnosis using DISCERN™ and CDP. •False Negative (FN) diagnosis: incremental cost of care avoidable with a correct AD diagnosis and appropriately directed medication. •True Positive (TP) diagnosis: AD medication costs; cost from a later TP diagnosis with the CDP versus DISCERN™ in the year of diagnosis, and savings from the delay in AD progression due to appropriate AD medication in patients who are correctly diagnosed after a FN diagnosis.•False Positive (FP) diagnosis: cost of AD medication for patients who do not have AD. A one-way sensitivity analysis was conducted to assess the effect of varying key clinical and cost parameters ±10%. An additional scenario analysis was developed to evaluate the impact of individual inputs. In the base scenario, DISCERN™ is estimated to decrease costs by $4.75M over three years, equating to approximately $63.11 saved per test per year for a cohort followed over three years. While the diagnosis cost is higher with DISCERN™ than with CDP modalities, this cost is offset by the higher overall costs associated with CDP due to the longer time needed to receive a TP diagnosis and the larger number of patients who receive a FN diagnosis and progress more rapidly than if they had received appropriate AD medication. The sensitivity analysis shows that the three parameters with the greatest impact on savings are: reduced sensitivity of DISCERN™, improved sensitivity of the CDP, and a reduction in the percentage of disease progression that is avoided with appropriate AD medication. A scenario analysis in which DISCERN™ reduces the utilization for patients of computed tomography from 21% in the base case to 16%, magnetic resonance imaging from 37% to 27% and cerebrospinal fluid biomarker testing, positive emission tomography, electroencephalograms, and polysomnography testing from 4%, 5%, 10%, and 8%, respectively, in the base case to 0%, results in an overall three-year net savings of $14.5M. DISCERN™ improves the rate of accurate, definitive diagnosis of AD earlier in the disease and may generate savings for Medicare Advantage plans.

Keywords: Alzheimer’s disease, budget, dementia, diagnosis.

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2 Aerobic Biodegradation of a Chlorinated Hydrocarbon by Bacillus Cereus 2479

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

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1 Redox-labeled Electrochemical Aptasensor Array for Single-cell Detection

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

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