Search results for: Chee Wai Woon

Authors: Seong Woon Lee, Soo Min Lim, Sung Sik Jeong, Jung Hoon Park

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The atmospheric concentration of greenhouse gas (GHG, Green House Gas) is increasing continuously as a result of the combustion of fossil fuels and industrial development. In response to this trend, many researches have been conducted on the reduction of GHG. Landfill gas (LFG, Land Fill Gas) is one of largest sources of GHG emissions containing the methane (CH₄) as a major constituent and can be considered renewable energy sources as well. In order to use LFG by connecting to the city pipe network, it required a process for removing impurities. In particular, oxygen must be removed because it can cause corrosion of pipes and engines. In this study, methane oxidation was used to eliminate oxygen from LFG and perovskite-type ceramic catalysts of La-Sr-Co-Fe-O composition was selected as a catalyst. Hollow fiber catalysts (HFC, Hollow Fiber Catalysts) have attracted attention as a new concept alternative because they have high specific surface area and mechanical strength compared to other types of catalysts. HFC was prepared by a phase-inversion/sintering technique using commercial La-Sr-Co-Fe-O powder. In order to measure the catalysts' activity, simulated LFG was used for feed gas and complete oxidation reaction of methane was confirmed. Pore structure of the HFC was confirmed by SEM image and perovskite structure of single phase was analyzed by XRD. In addition, TPR analysis was performed to verify the oxygen adsorption mechanism of the HFC. Acknowledgement—The project is supported by the ‘Global Top Environment R&D Program’ in the ‘R&D Center for reduction of Non-CO₂ Greenhouse gases’ (Development and demonstration of oxygen removal technology of landfill gas) funded by Korea Ministry of Environment (ME).

Keywords: complete oxidation, greenhouse gas, hollow fiber catalyst, land fill gas, oxygen removal, perovskite catalyst

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Authors: Koon Weng Lau, Chee Dhang Chen, Abdul Aziz Azidah, Mohd Sofian-Azirun

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Recently, Sarawak state of Malaysia encounter an outbreak of dengue fever. Aedes albopictus has incriminated as one of the important vectors of dengue transmission. Without an effective vaccine, approaches to control or prevent dengue will be a focus on the vectors. The control of Aedes mosquitoes is still dependent on the use of chemical insecticides and insecticide resistance represents a threat to the effectiveness of vector control. This study was conducted to determine the resistance status of 11 active ingredients representing four major insecticide classes: DDT, dieldrin, malathion, fenitrothion, bendiocarb, propoxur, etofenprox, deltamethrin, lambda-cyhalothrin, cyfluthrin, and permethrin. Standard WHO test procedures were conducted to determine the insecticide susceptibility. Aedes albopictus collected from Kapit (resistance ratio, RR = 1.04–3.02), Kuching (RR = 1.17–4.61), and Sibu (RR = 1.06–3.59) exhibited low resistance toward all insecticides except dieldrin. This study reveled that dieldrin is still effective against Ae. albopictus, followed by fenitrothion, cyfluthrin, and deltamethrin. In conclusion, Ae. albopictus in Sarawak exhibited different resistance levels toward various insecticides and alternative solutions should be implemented to prevent further deterioration of the condition.

Keywords: Aedes albopictus, dengue, insecticide resistance, Malaysia

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Authors: Santiranjan Shannigrahi, Mohit Sharma, Ivan Tan Chee Kiang, Yong Anna Marie

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Polymer-ceramics composites are gaining importance due to their high specific strength, corrosion resistance, and high mechanical properties, as well as low cost. As a result, polymer composites are suitable for various industrial applications, like automobiles, aerospace, and biomedical areas. The present work comprises the development of polymer-ceramic composite films and is tested for the harsh environment including weatherability and UV barrier property. The polymer composite films are kept in weather chamber for a fixed period of time followed by tested for their physical, mechanical and chemical properties. The composite films are fabricated using compounding followed by hot pressing. UV-visible spectroscopy results reveal that the pure polymer polyethylene (PE) films are transparent in the visible range and do not absorb UV. However, polymer ceramic composite films start absorbing UV completely even at very low filler loading amount of 5 wt.%. The changes in tensile properties of the various composite films before and after UV illuminations for 40 hrs at 60 degC are analyzed. The tensile strength of neat PE film has been observed 8% reduction, whereas the remarkable increase in tensile strength has been observed (18% improvement for 10 wt. % filled composites films). The UV exposure leads to strengthen the crosslinking among PE polymer chains in the filled composite films, which contributes towards the incremented tensile strength properties.

Keywords: polymer ceramic composite, processing, harsh environment, mechanical properties

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Authors: Vivien Chee Pei Wei

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Today we live in a scenario in which letters share space with images on screens that vary in size, shape, and style. The popularization of television, then the computer and now the e-readers, tablets, and smartphones made the electronic assume the role that previously was restricted to printed materials. Since the extensive use of new technologies to produce, disseminate, collect and access electronic publications began, the changes to reading has been intensified. To be able to read online, it involves more than just utilizing specific skills, strategies, and practices, but also in negotiating multiple information sources. In this study, different perspectives of digital reading are being explored in order to define the key aspects of the term. The focus is to explore how new technologies affect how undergraduates’ reading behavior, which in turn, gives readers different reading levels and engagement with the text and other support materials in the same media. There is also the importance of the relationship between reading platforms, reading levels and formats of electronic publications. The study looks at the online reading practices of about 100 undergraduates from a local university. The data collected using the survey and interviews with the respondents are analyzed thematically. Findings from this study found that both digital and traditional reading are interrelated, and should not be viewed as separate, but complementary to each other. However, reading online complicates some of the skills required by traditional reading. Consequently, in order to successfully read and comprehend multiple sources of information online, undergraduates need regular opportunities to practice and develop their skills as part of their natural reading practices.

Keywords: concepts, digital reading, literacy, traditional reading

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Authors: Samson Olasunkanmi Oluga, Teh Chee Send, Gerard Sagaya Raj Rajo

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It is ironical that the national anthems of many nations that are in the forefront of the global condemnation of violence of all forms have portions or expressions that propagate various forms of linguistic violence which advocate attacking opponents, going to war, shedding blood and sacrificing lives. These diametrically contradict contemporary yearnings for global tranquility and the ideals of the United Nations established for the maintenance of international peace and harmony aimed at making the world a safe haven for all and sundry. The linguistic violence of many national anthems is manipulatively constructed /presented via the instrumentality of the figurative or rhetorical language. This helps to linguistically embellish the violent ideas communicated and makes them sound somehow better or logical to the target audience with the intention of cognitively manipulating them to accept or rationalize such violent ideas. This paper, therefore, presents the outcome of a linguistic exploration/examination of national anthems which reveals elements or cases manipulative figurative linguistic violence in the anthems of twenty-one (21) nations. The paper details a Socio-Cognitive Critical Discourse Analysis of the manipulative figures of comparison, contrast, indirectness, association and sound used to convey the linguistic violence of the identified national anthems. Finally, the paper advocates the need for linguistic overhaul of affected anthems so that the language of anthems which epitomize nations can be pacific and in tandem with contemporary global trends.

Keywords: national anthems, linguistic violence, figurative language, cognitive, manipulation, CDA

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Authors: Feng Ji Mervin Goh, Edmund Chee Jian Pua, Stuart Alexander Cook

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Dilated cardiomyopathy (DCM) is a common cause of heart failure. Genetic mutations account for 50% of DCM cases with TTN mutations being the most common, accounting for up to 25% of DCM cases. However, the genetic architecture underlying Asian DCM patients is unknown. We evaluated 68 patients (female= 17) with DCM who underwent follow-up at the National Heart Centre, Singapore from 2013 through 2014. Clinical data were obtained and analyzed retrospectively. Genomic DNA was subjected to next-generation targeted sequencing. Nextera Rapid Capture Enrichment was used to capture the exons of a panel of 169 cardiac genes. DNA libraries were sequenced as paired-end 150-bp reads on Illumina MiSeq. Raw sequence reads were processed and analysed using standard bioinformatics techniques. The average age of onset of DCM was 46.1±10.21 years old. The average left ventricular ejection fraction (LVEF), left ventricular diastolic internal diameter (LVIDd), left ventricular systolic internal diameter (LVIDs) were 26.1±11.2%, 6.20±0.83cm, and 5.23±0.92cm respectively. The frequencies of mutations in major DCM-associated genes were as follows TTN (5.88% vs published frequency of 20%), LMNA (4.41% vs 6%), MYH7 (5.88% vs 4%), MYH6 (5.88% vs 4%), and SCN5a (4.41% vs 3%). The average callability at 10 times coverage of each major gene were: TTN (99.7%), LMNA (87.1%), MYH7 (94.8%), MYH6 (95.5%), and SCN5a (94.3%). In conclusion, TTN mutations are not common in Singaporean DCM patients. The frequencies of other major DCM-associated genes are comparable to frequencies published in the current literature.

Keywords: heart failure, dilated cardiomyopathy, genetics, next-generation sequencing

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Authors: Chee Dhang Chen, Hiroyuki Takaoka, Koon Weng Lau, Poh Ruey Tan, Ai Chdon Chin, Van Lun Low, Abdul Aziz Azidah, Mohd Sofian-Azirun

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Susceptibility status of Simulium (Simulium) nobile (Diptera: Simuliidae) adults obtained from Pahang, Malaysia was evaluated against 11 adulticides representing four major insecticide classes: organochlorines (DDT, dieldrin), organophosphates (malathion, fenitrothion), carbamates (bendiocarb, propoxur) and pyrethroids (etofenprox, deltamethrin, lambdacyhalothrin, permethrin, cyfluthrin). The adult bioassay was conducted according to WHO standard protocol to determine the insecticide susceptibility. Mortality at 24 h post treatment was used as indicator for susceptibility status. The results revealed that S. nobile obtained was susceptible to propoxur, cyfluthrin and bendiocarb with 100% mortality. S. nobile was resistant or exhibited some tolerant against lambdacyhalothrin and deltamethrin with mortality ranged ≥ 90% but < 98%. S. nobile populations in Pahang exhibited different level of resistant against 11 adulticides with mortality ranged from 60.00 ± 10.00 to 100.00 ± 0.00. In conclusion, S. nobile populations in Pahang were susceptible to propoxur, cyfluthrin and bendiocarb. The susceptibility status of S. nobile in descending order was propoxur, cyfluthrin > bendicarb > deltamethrin > lambdacyhalothrin > permethrin > etofenprox > DDT > malathion > fenitrothion > dieldrin. Regular surveys should be conducted to monitor the susceptibility status of this insect vector in order to prevent further development of resistance.

Keywords: black fly, adult bioassay, insecticide resistance, Malaysia

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Authors: Nordin Mamat, Abdul Rahim Razalli, Loy Chee Luen, Abdul Talib Hashim

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This study was designed to investigate preschool environment in multi-ethnic preschool in Malaysia. The objectives are to identify the quality of work environment in multi-ethnic preschools; to investigate the practices of teachers’ role and responsibility; and to identify the quality of human resources. The study involved 2004 respondents who are the staff of multi-ethnic preschool from the government agency who provide preschool service. This study was conducted using a mixed method in which questionnaires and interviews were used to obtain data from respondents. The findings were analysed using mean and used Likert scale to determine the three-stage level such as the high, moderate and low. Findings indicated that the work environment at a moderate level, but the facilities provided insufficient to carry out educational activities with children. The result based on ranking of duties and responsibilities of teachers in multi-ethnic preschool shows the teachers practice daily record of children's development is very little, that only 65 persons are recording the child's development. The poor ratio of teachers and child in multi-ethnic preschool is between 25 to 35 children per class which means the children need a lot of attention. Meanwhile, the work environment is moderate with a mean score of 3.65 and overall mean score for level of staff career development 3.66 also moderate. The findings indicate the facilities provided in their workplace and staff career development requires improvements. Overall, the level of work environment is moderate, and it needs an improvement in term of facilities.

Keywords: environment, human resources, multi-ethnic preschool, quality teacher

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Authors: Jin Hun Choi, So Hyun Ahn, Seong Keun Wang, Ik-Seung Chee, Jung Lan Kim, Sun Woo Lee

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Objectives: The purpose of the study is to investigate the effects of day hospital care in hospitalized schizophrenic patients in terms of treatment adherence and treatment outcomes. Methods: Among schizophrenic patients hospitalized between 2011 and 2012, 23 day hospital care patient and 40 control subjects were included in the study. All candidates underwent Beck Cognitive Insight Scale, Drug Attitude Inventory, World Health Organization Quality of Life Assessment and Psychological Well-Being Scale when their symptoms were stabilized during hospitalization, and after being discharged, 23 patients received day hospital care for two months and then changed to out-patient care while 40 patients received out-patient care immediately after discharge. At the point of two months of out-patient care, the treatment adherence of the two groups was evaluated; tracking observation was performed until February, 2013, and survival rates were compared between the two groups. Results: Treatment adherence was higher in the day hospital care group than in the control group. Kaplan-Meier survival analysis showed a higher survival rate for the day hospital care group compared to the control group. Levels of cognitive insight and quality of life were higher after day hospital care than before day hospital care in the day hospital care group. Conclusions: Through the study, it was confirmed that when hospitalized schizophrenic patients received continuous day hospital care after being discharged, they received further out-patient care more faithfully. The study is considered to aid in the understanding regarding schizophrenic patients’ treatment adherence issues and improvement of treatment outcomes.

Keywords: schizophrenia, day hospital care, adherence, outcomes

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Authors: Hyun Chang Kim, Yong Hun Jung, Kyung Woon Jeung

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Background: From the viewpoint of cardiac pump theory, the area of the left ventricle (LV) subjected to compression increases as the LV lies closer to the sternum, possibly resulting in higher blood flow in patients with LV closer to the sternum. However, no study has evaluated LV position during cardiac arrest or its relationship with hemodynamic parameters during cardiopulmonary resuscitation (CPR). The objectives of this study were to determine whether the position of the LV relative to the anterior-posterior axis representing the direction of chest compression shifts during cardiac arrest and to examine the relationship between LV position and hemodynamic parameters during CPR. Methods: Subcostal view echocardiograms were obtained from 15 pigs with the transducer parallel to the long axis of the sternum before inducing ventricular fibrillation (VF) and during cardiac arrest. Computed tomography was performed in three pigs to objectively observe LV position during cardiac arrest. LV position parameters including the shortest distance between the anterior-posterior axis and the mid-point of the LV chamber (DAP-MidLV), the shortest distance between the anterior-posterior axis and the LV apex (DAP-Apex), and the area fraction of the LV located on the right side of the anterior-posterior axis (LVARight/LVATotal) were measured. Results: DAP-MidLV, DAP-Apex, and LVARight/LVATotal decreased progressively during untreated VF and basic life support (BLS), and then increased during advanced cardiovascular life support (ACLS). A repeated measures analysis of variance revealed significant time effects for these parameters. During BLS, the end-tidal carbon dioxide and systolic right atrial pressure were significantly correlated with the LV position parameters. During ACLS, systolic arterial pressure and systolic right atrial pressure were significantly correlated with DAP-MidLV and DAP-Apex. Conclusions: LV position changed significantly during cardiac arrest compared to the pre-arrest baseline. LV position during CPR had significant correlations with hemodynamic parameters.

Keywords: heart arrest, cardiopulmonary resuscitation, heart ventricle, hemodynamics

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Authors: Hadri Hussain, M.Nasir Ibrahim, Chee-Ming Ting, Mariani Idroas, Fuad Numan, Alias Mohd Noor

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A biometric security system has become an important application in client identification and verification system. A conventional biometric system is normally based on unimodal biometric that depends on either behavioural or physiological information for authentication purposes. The behavioural biometric depends on human body biometric signal (such as speech) and biosignal biometric (such as electrocardiogram (ECG) and phonocardiogram or heart sound (HS)). The speech signal is commonly used in a recognition system in biometric, while the ECG and the HS have been used to identify a person’s diseases uniquely related to its cluster. However, the conventional biometric system is liable to spoof attack that will affect the performance of the system. Therefore, a multimodal biometric security system is developed, which is based on biometric signal of ECG, HS, and speech. The biosignal data involved in the biometric system is initially segmented, with each segment Mel Frequency Cepstral Coefficients (MFCC) method is exploited for extracting the feature. The Hidden Markov Model (HMM) is used to model the client and to classify the unknown input with respect to the modal. The recognition system involved training and testing session that is known as client identification (CID). In this project, twenty clients are tested with the developed system. The best overall performance at 44 kHz was 93.92% for ECG and the worst overall performance was ECG at 88.47%. The results were compared to the best overall performance at 44 kHz for (20clients) to increment of clients, which was 90.00% for HS and the worst overall performance falls at ECG at 79.91%. It can be concluded that the difference multimodal biometric has a substantial effect on performance of the biometric system and with the increment of data, even with higher frequency sampling, the performance still decreased slightly as predicted.

Keywords: electrocardiogram, phonocardiogram, hidden markov model, mel frequency cepstral coeffiecients, client identification

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Authors: Siew Ming Thang, Wong Hoo Keat, Chee Hao Sue, Fung Lan Loo, Ahju Rosalind

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There are numerous studies that explored young learners’ literacy skills in Malaysia but none that uses the eye-tracking device to track their cognitive processes when reading picture storybooks. This study used this method to investigate two groups of prereaders’ cognitive processes in four conditions. (1) A congruent picture was presented, and a matching narration was read aloud by a recorder; (2) Children heard a narration telling about the same characters in the picture but involves a different scene; (3) Only a picture with matching text was present; (4) Students only heard the reading aloud of the text on the screen. The two main objectives of this project are to test which content of pictures helps the prereaders (i.e., young children who have not received any formal reading instruction) understand the narration and whether children try to create a coherent mental representation from the oral narration and the pictures. The study compares two groups of children from two different kindergartens. Group1: 15 Chinese children; Group2: 17 Malay children. The medium of instruction was English. An eye-tracker were used to identify Areas of Interest (AOI) of each picture and the five target elements and calculate number of fixations and total time spent on fixation of pictures and written texts. Two mixed factorial ANOVAs with the storytelling performance (good, average, or weak) and vocabulary level (low, medium, high) as between-subject variables, and the Areas of Interests (AOIs) and display conditions as the within-subject variables were performedon the variables.

Keywords: eye-tracking, cognitive processes, literacy skills, prereaders, visual attention

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Authors: Cher Rong Matthew Kong, Edmund Tian, Seng Poon Ong, Chee Sian Gan

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Gamma-aminobutyric acid (GABA) is a non-protein amino acid that is a functional constituent of certain rice varieties. When consumed, it decreases blood pressure and reduces the risk of hypertension-related diseases. This has led to more research dedicated towards the development of functional food products (e.g. germinated brown rice) with enhanced GABA content, and the development of these functional food products has led to increased demand for instrument-based methods that can efficiently and effectively determine GABA content. Current analytical methods require analyte derivatisation, and have significant disadvantages such as being labour intensive and time-consuming, and being subject to analyte loss due to the increased complexity of the sample preparation process. To address this, an LC-MS-MS method for the determination of GABA in rice products has been developed and validated. This developed method involves a relatively simple sample preparation process before analysis using HILIC LC-MS-MS. This method eliminates the need for derivatisation, thereby significantly reducing the labour and time associated with such an analysis. Using LC-MS-MS also allows for better differentiation of GABA from any potential co-eluting compounds in the sample matrix. Results obtained from the developed method demonstrated high linearity, accuracy, and precision for the determination of GABA (1ng/L to 8ng/L) in a variety of brown rice products. The method can significantly simplify sample preparation steps, improve the accuracy of quantitation, and increase the throughput of analyses, thereby providing a quick but effective alternative to established instrumental analysis methods for GABA in rice.

Keywords: functional food, gamma-aminobutyric acid, germinated brown rice, method development

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Authors: Ji Young Lee, Jong Woon Kim

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It is essential to secure high availability of railway vehicles to realize high quality and efficiency of railway service. Once the availability decreased, planned railway service could not be provided or more cars need to be reserved. additional cars need to be purchased or the frequency of railway service could be decreased. Such situation would be a big loss in terms of quality and cost related to railway service. Therefore, we make various efforts to get high availability of railway vehicles. Because it is a big loss to operators, we make various efforts to get high availability of railway vehicles. To secure high availability, the idle time of the vehicle needs to be reduced and the following methods are applied to railway vehicles. First, through modularization design, exchange time for line replaceable units is reduced which makes railway vehicles could be put into the service quickly. Second, to reduce periodic preventive maintenance time, preventive maintenance with short period would be proceeded test oriented to minimize the maintenance time, and reliability is secured through overhauls for each main component. With such design changes for railway vehicles, modularized components are exchanged first at the time of vehicle failure or overhaul so that vehicles could be put into the service quickly and exchanged components are repaired or overhauled. Therefore, spare components are required for any future failures or overhauls. And, as components are modularized and costs for components are high, it is considerably important to get reasonable quantities of spare components. Especially, when a number of railway vehicles were put into the service simultaneously, the time of overhauls come almost at the same time. Thus, for some vehicles, components need to be exchanged and overhauled before appointed overhaul period so that these components could be secured as spare parts for the next vehicle’s component overhaul. For this reason, components overhaul time and spare parts quantities should be decided at the same time. This study deals with the time of overhauls for repairable components of railway vehicles and the calculation of spare parts quantities in consideration of future failure/overhauls. However, as railway vehicles are used according to the service schedule, maintenance work cannot be proceeded after the service was closed thus it is quite difficult to resolve this situation mathematically. In this study, Simulation software system is used in this study for analyzing the time of overhauls for repairable components of railway vehicles and the spare parts for the railway systems.

Keywords: overhaul time, rolling stocks, simulation, spare parts

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Authors: Lim Chen Kim, Tan Kian Lam, Chan Yi Chee

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This research presents a multi-modal simulation in the reconstruction of the past and the construction of present in digital cultural heritage on mobile platform. In bringing the present life, the virtual environment is generated through a presented scheme for rapid and efficient construction of 360° panoramic view. Then, acoustical heritage model and crowd model are presented and improvised into the 360° panoramic view. For the reconstruction of past life, the crowd is simulated and rendered in an old trading port. However, the keystone of this research is in a virtual walkthrough that shows the virtual present life in 2D and virtual past life in 3D, both in an environment of virtual heritage sites in George Town through mobile device. Firstly, the 2D crowd is modelled and simulated using OpenGL ES 1.1 on mobile platform. The 2D crowd is used to portray the present life in 360° panoramic view of a virtual heritage environment based on the extension of Newtonian Laws. Secondly, the 2D crowd is animated and rendered into 3D with improved variety and incorporated into the virtual past life using Unity3D Game Engine. The behaviours of the 3D models are then simulated based on the enhancement of the classical model of Boid algorithm. Finally, a demonstration system is developed and integrated with the models, techniques and algorithms of this research. The virtual walkthrough is demonstrated to a group of respondents and is evaluated through the user-centred evaluation by navigating around the demonstration system. The results of the evaluation based on the questionnaires have shown that the presented virtual walkthrough has been successfully deployed through a multi-modal simulation and such a virtual walkthrough would be particularly useful in a virtual tour and virtual museum applications.

Keywords: Boid Algorithm, Crowd Simulation, Mobile Platform, Newtonian Laws, Virtual Heritage

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Authors: Chee Fei Chang, May Yee Ng

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As a result of its geographical location, flood is a natural disaster that happens regularly in Malaysia. Every year, heavy rainfall is brought by the cyclical monsoon to the East coast of Peninsular Malaysia. In recent years, the occurrence of unexpected heavy downpour somehow connected to climate-change phenomena is also on the increasing trend. Ironically, despite that Malaysians have suffered significant monetary losses as a result of the recurring floods in past many decades, little has been done by the government from the perspective of taxation. Perhaps due to political reason or as a populist measure, the federal and local government are more inclined to offer small cash handout then rolling out long-term tax policy or measure in relieving the financial and tax burden of the victims and affected business entities. Except for the one-off tax break granted to affected businesses in 2007, the authors have not found any income tax exemption or deduction order gazetted with regard to flood disaster. Hence, it is imperative for this study to explore the need and challenges of implementing flood inflicted disaster tax relief or credit in Malaysia. This research consists of two major parts. First, the assessment of relevant tax policies/ measures with regard to non-government organisations and other affected parties. Content and thematic analyses will be applied on current tax legislations and orders issued for this part. Second, a comparative analysis will be conducted benchmarking various disaster tax reliefs and credits implemented in developed countries. Resulting from the increasing climate change-related disasters in Malaysia, the findings of this study will shed light on the importance of introducing disaster tax relief measures to assist individual victims as well as the affected businesses.

Keywords: climate-changed related disaster, disaster tax credits, tax relief for victims, tax measures for disaster recovery

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Authors: Kun Hua O., Dong Woon Kim, Won Hyung Lee

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Introduction: Neuropathic pain induced by spinal or peripheral nerve injury is highly resistant to common painkillers, nerve blocks, and other pain management approaches. Recently, several new therapeutic drug candidates have been developed to control neuropathic pain. In this study, we used the spinal nerve L5 ligation (SNL) model to investigate the ability of intrathecal or intravenous Evans blue to decrease pain behavior and to study the relationship between Evans blue and the neural structure of pain transmission. Method: Neuropathic pain (allodynia) of the left hind paw was induced by unilateral SNL in Sprague-Dawley rats(n=10) in each group. Evans blue (5, 15, 50μg/10μl) or phosphate buffer saline(PBS,10μl) was injected intrathecally at 3days post-ligation or intravenously(1mg/200 μl) 3days and 5days post-ligation . Mechanical sensitivity was assessed using Von Frey filaments at 3 days post-ligation and at 2 hours, days 1, 2, 3, 5,7 after intrathecal Evans blue injection, and on days 2, 4, 7, and 11 at 14 days after intravenous injection. In the intrathecal group, microglia and glutaminergic neurons in the dorsal horn and VNUT(vesicular nucleotide transporter) in the dorsal root ganglia were tested to evaluate co-staining with Evans blue. The experimental procedures were performed in accordance with the animal care guideline of the Korean Academy of Medical Science(Animal ethic committee of Chungnam National University Hospital: CNUH-014-A0005-1). Results: Tight ligation of the L5 spinal nerve induced allodynia in the left hind paw 3 days post-ligation. Intrathecal Evans blue most significantly(P＜0.001) alleviated allodynia at 2 days after intrathecal, but not an intravenous injection. Glutaminergic neurons in the dorsal horn and VNUT in the dorsal root ganglia were co-stained with Evans blue. On the other hand, microglia in the dorsal horn were partially co-stained with Evans blue. Conclusion: We confirmed that Evans blue might have an analgesic effect through the central nervous system, not another system in neuropathic pain of the SNL animal model. These results suggest Evans blue may be a potential new drug for the treatment of chronic pain. This research was supported by the National Research Foundation of Korea (NRF-2020R1A2C100757512), funded by the Ministry of Education.

Keywords: neuropathic pain, Evas blue, intrathecal, intravenous

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Authors: Betsy Ng, Seng Chee Tan, Choon Lang Quek, Peter Looker, Jaime Koh

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The purpose of the present study was to determine the particular scale rating that stands out for an instrument. The instrument was designed to assess student perceptions of various learning environments, namely face-to-face, online and blended. The original instrument had a 5-point Likert items (1 = strongly disagree and 5 = strongly agree). Alternate versions were modified with a 6-point Likert scale and a bar scale rating. Participants consisted of undergraduates in a local university were involved in the usability testing of the instrument in an electronic setting. They were presented with the 5-point, 6-point and percentage-bar (100-point) scale ratings, in response to their perceptions of learning environments. The 5-point and 6-point Likert scales were presented in the form of radio button controls for each number, while the percentage-bar scale was presented with a sliding selection. Among these responses, 6-point Likert scale emerged to be the best overall. When participants were confronted with the 5-point items, they either chose 3 or 4, suggesting that data loss could occur due to the insensitivity of instrument. The insensitivity of instrument could be due to the discreet options, as evidenced by response interpolation. To avoid the constraint of discreet options, the percentage-bar scale rating was tested, but the participant responses were not well-interpolated. The bar scale might have provided a variety of responses without a constraint of a set of categorical options, but it seemed to reflect a lack of perceived and objective accuracy. The 6-point Likert scale was more likely to reflect a respondent’s perceived and objective accuracy as well as higher sensitivity. This finding supported the conclusion that 6-point Likert items provided a more accurate measure of the participant’s evaluation. The 5-point and bar scale ratings might not be accurately measuring the participants’ responses. This study highlighted the importance of the respondent’s perception of accuracy, respondent’s true evaluation, and the scale’s ease of use. Implications and limitations of this study were also discussed.

Keywords: usability, interpolation, sensitivity, Likert scales, accuracy

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Authors: Asfa Hussain, Chee Tang, Mien Nguyen

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Introduction: The safe usage of medications is dependent on clear, well-documented prescribing. Medical drug charts should be regularly checked to ensure that they are fit for purpose. Aims: The purpose of this study was to evaluate whether the Isabel Hospice drug charts were effective or prone to medical errors. The aim was to create a comprehensive palliative care drug chart in line with medico-legal guidelines and to minimise drug administration and prescription errors. Methodology: 50 medical drug charts were audited from March to April 2020, to assess whether they complied with medico-legal guidelines, in a hospice within East of England. Meetings were held with the larger multi-disciplinary team (MDT), including the pharmacists, nursing staff and doctors, to raise awareness of the issue. A preliminary drug chart was created, using the input from the wider MDT. The chart was revised and trialled over 15 times, and each time feedback from the MDT was incorporated into the subsequent template. In the midst of the COVID-19 pandemic in September 2020, the finalised drug chart was trialled. 50 new palliative drug charts were re-audited, to evaluate the changes made. Results: Prescribing and administration errors were high prior to the implementation of the new chart. This improved significantly after introducing the new drug charts, therefore improving patient safety and care. The percentage of inadequately documented allergies went down from 66% to 20% and incorrect oxygen prescription from 40% to 16%. The prescription drug-drug interactions decreased by 30%. Conclusion: It is vital to have clear standardised drug charts, in line with medico-legal standards, to allow ease of prescription and administration of medications and ensure optimum patient-centred care. This closed loop audit demonstrated significant improvement in documentation and prevention of possible fatal drug errors and interactions.

Keywords: palliative care, drug chart, medication errors, drug-drug interactions, COVID-19, patient safety

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Authors: Ralene Sim, Stefan Mueller, N. Gopalakrishna Iyer, Ngian Chye Tan, Khee Chee Soo, R. Shetty Mahalakshmi, Hiang Khoon Tan

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Background: Regional failure in nasopharyngeal carcinoma (NPC) is managed by salvage treatment in the form of neck dissection. Radical neck dissection (RND) is preferred over modified radical neck dissection (MRND) since it is traditionally believed to offer better long-term disease control. However, with the advent of more advanced imaging modalities like high-resolution Magnetic Resonance Imaging, Computed Tomography, and Positron Emission Tomography-CT scans, earlier detection is achieved. Additionally, concurrent chemotherapy also contributes to reduced tumour burden. Hence, there may be a lesser need for an RND and a greater role for MRND. With this retrospective study, the primary aim is to ascertain whether MRND, as opposed to RND, has similar outcomes and hence, whether there would be more grounds to offer a less aggressive procedure to achieve lower patient morbidity. Methods: This is a retrospective study of 66 NPC patients treated at Singapore General Hospital between 1994 to 2016 for histologically proven regional recurrence, of which 41 patients underwent RND and 25 who underwent MRND, based on surgeon preference. The type of ND performed, primary treatment mode, adjuvant treatment, and pattern of recurrence were reviewed. Overall survival (OS) was calculated using Kaplan-Meier estimate and compared. Results: Overall, the disease parameters such as nodal involvement and extranodal extension were comparable between the two groups. Comparing MRND and RND, the median (IQR) OS is 1.76 (0.58 to 3.49) and 2.41 (0.78 to 4.11) respectively. However, the p-value found is 0.5301 and hence not statistically significant. Conclusion: RND is more aggressive and has been associated with greater morbidity. Hence, with similar outcomes, MRND could be an alternative salvage procedure for regional failure in selected NPC patients, allowing similar salvage rates with lesser mortality and morbidity.

Keywords: nasopharyngeal carcinoma, neck dissection, modified neck dissection, radical neck dissection

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Authors: Sofiya An, Akbota Kanderzhanova, Assel Akhmetova, Faye Foster, Chee K. Chan

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Healthcare, education and social support for children with autism in Kazakhstan has been evolving and transforming over the last three decades. There is still limited knowledge of the use of complementary and alternative medicine by families caring for autistic children in this post-Soviet region. An exploratory qualitative focus group study of Kazakhstani families was carried out to capture and understand their experiences of using complementary and alternative (CAM) medicine. A total of six focus groups were conducted in five cities across the country including Nur-Sultan, Almaty, Kyzylorda, Karaganda and Taraz. The perceived factors driving the availability, choice, and use of complementary and alternative medicine by families of autistic children in the country were distilled and evaluated. The data collected was analyzed using a framework analysis and themes and subthemes were developed. Two major themes stood out. The first was the “unmet needs”, which relates to the predisposing factors that motivate parents to CAM uptake, and the second was the “chasing hope”, which relates to the enabling factors that facilitate parents’ uptake of CAM. Fear of missing out (FOMO) is a latent underlying motivation underscoring these two themes as well. Parents of autism spectrum disorder (ASD) children in Kazakhstan have to deal with many challenges when seeking treatment for their children with ASD. They are prepared and resort to try out whatever CAM interventions available. The motivation and rationale of choice of use is driven by the lack of options and the hope of any potential positive outcome rather than from rational decisions based on efficacy or the evidence-based data of CAM. Parents get desperate and are willing to try CAM regardless of and independent of their cultural and belief systems and they do not want to miss out just in case it might work. This study also gives an international and cross-cultural perspective on the motives, choice and practice of parents with ASD children using CAM in Kazakhstan, a Central Asian country.

Keywords: autism spectrum disorder, Central Asia, complementary and alternative medicine, cross-cultural perspective, qualitative research

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Authors: Lynn Louis, Bor Shin Chee, Marion McAfee, Michael Nugent

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This article aims to develop a sustained release formulation of microspheres containing the mTOR inhibitor Everolimus (EVR) using Polyvinyl alcohol (PVA) to enhance the bioavailability of the drug and to overcome poor solubility characteristics of Everolimus. This paper builds on recent work in the manufacture of microspheres using the sessile droplet technique by freezing the polymer-drug solution by suspending the droplets into pre-cooled ethanol vials immersed in liquid nitrogen. The spheres were subjected to 6 freezing cycles and 3 freezing cycles with thawing to obtain proper geometry, prevent aggregation, and achieve physical cross-linking. The prepared microspheres were characterised for surface morphology by SEM, where a 3-D porous structure was observed. The in vitro release studies showed a 62.17% release over 12.5 days, indicating a sustained release due to good encapsulation. This result is comparatively much more than the 49.06% release achieved within 4 hours from the solvent cast Everolimus film as a control with no freeze-thaw cycles performed. The solvent cast films were made in this work for comparison. A prolonged release of Everolimus using a polymer-based drug delivery system is essential to reach optimal therapeutic concentrations in treating SEGA tumours without systemic exposure. These results suggest that the combination of PVA and Everolimus via a rheological synergism enhanced the bioavailability of the hydrophobic drug Everolimus. Physical-chemical characterisation using DSC and FTIR analysis showed compatibility of the drug with the polymer, and the stability of the drug was maintained owing to the high molecular weight of the PVA. The obtained results indicate that the developed PVA/EVR microsphere is highly suitable as a potential drug delivery system with improved bioavailability in treating Subependymal Giant cell astrocytoma (SEGA).

Keywords: drug delivery system, everolimus, freeze-thaw cycles, polyvinyl alcohol

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Authors: Chee Young Kim, Tae Hyun Kim, Anbok Lee, Hyun-Ah Kim, Woosung Lim, Ku Sang Kim, Jinsun Lee, Yoo Seok Kim, Beom Seok Ko

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Background: CollatampTM is Gentamicin-containing collagen sponge well known for its hemostatic effect, commonly utilized in surgeries. We inserted CollatempTM wrapped by SurgicelTM (oxidized cellulose polymer) to fill up the defect after breast conserving surgery. The purpose of this study is to verify the furthermore cosmetic value of CollatampTM in breast conserving surgery conducted in breast cancer patients. Methods: 17 patients were enrolled in this study, underwent breast conserving surgery with CollatampTM wrapped by SurgicelTM insertion, in Inje University Busan Paik Hospital from October 2015 to September 2016. Patient satisfaction, cosmetic outcome, results at 6 months from operation was analyzed to verify the effectiveness and usefulness of CollatampTM for cosmetics. Patient satisfaction was investigated through interviews on a scale of good, fair, poor, and the cosmetic outcome was investigated through physical examination by a surgeon who did not participate in the operations. Results: Among 17 patients, nine of them gave ‘good’ for patient satisfaction, eight gave ‘fair’ and none of them ‘poor’. Also, cosmetic outcome came out with 11 ‘good’s, six ‘fair’s, no ‘poor’. In ‘good’ patient satisfaction group, the mean value of resection to breast volume ratio was 16%, compared to 24% of ‘fair’ group. The mean value of actual resection volume was 100.6cm3, 102.7cm3 each. In ‘good’ cosmetic outcome group, the mean value of resection to breast volume ratio was 18%, compared to 23% of ‘fair’ group. The mean value of actual resection volume was 99.2cm3, 105.9cm3 respectively. According to these results, patient satisfaction and cosmetic outcome after surgeries were more reliable on the resection to breast volume ratio, rather than the actual resection volume. There were eight cases of postoperative complications, consisting of a lymphedema, a seroma, and six patients had mild pain. Conclusions: Cosmetic effect of CollatampTM in breast conserving surgery was more reliable on the resection to breast volume ratio, rather than the actual resection volume. In this short term survey, patients were tend to be satisfied with the cosmetics, all giving either good or fair scores. However, long term outcomes should be further assessed.

Keywords: breast cancer, breast conserving surgery, collatamp, cosmetics

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Authors: Joseph Shanks, Matthew Taylor, Foong Kiew Ooi, Chee Keong Chen

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Introduction: Evidences of relationship between bone, muscle and standard of competition among young British population is limited in literature. The current literature recognises the independent and synergistic effects of fat free and fat mass as the stimulus for osteogenesis. This study assessed the extent to which physical activity (PA) and standard of competition (CS) influences quantitative ultrasound (QUS) measurements of bone on a cross-sectional basis accounting for muscular strength and anthropometrics in British young males. Methods: Pre-screening grouped 66 males aged 18-25 years into controls (n=33) and district level athletes (DLAs) (n=33) as well as low (n=21), moderate (n=23) and high (n=22) physical activity categories (PACs). All participants underwent QUS measurements of bone (4 sites, i.e. dominant distal radius (DR), dominant mid-shaft tibia (DT), non-dominant distal radius (NR) and non-dominant mid-shaft tibia (NT)), isokinetic strength tests (dominant and non-dominant knee flexion and extension) and anthropometric measurements. Results: There were no significant differences between any of the groups with respect to QUS measurements of bone at all sites with regards to PACs or CS. Significant higher isokinetic strength values were observed in DLAs than controls (p < 0.05), and higher than low PACs (p < 0.05) at 60o.s-1 of concentric and eccentric measurements. No differences in subcutaneous fat thickness were found between all the groups (CS or PACs). Percentages of body fat were significantly higher (p < .05) in low than high PACs and CS groups. There were significant positive relationships between non dominant radial speed of sound and fat free mass at both DR (r=0.383, p=0.001) and NR (r=0.319, p=0.009) sites in all participants. Conclusion: The present study findings indicated that muscular strength and body fat are closely related to physical activity level and standard of competition. However, bone health status reflected by quantitative ultrasound (QUS) measurements of bone is not related to physical activity level and standard of competition in British young males.

Keywords: bone, muscular strength, physical activity, standard of competition

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Authors: Foong Kiew Ooi, Aidi Naim Mohamad Samsani, Chee Keong Chen, Mohamed Saat Ismail

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Introduction: Prolonged exercise in a hot and humid environment can result in glycogen depletion and associated with loss of body fluid. Carbohydrate contained in sports beverages is beneficial for improving sports performance and preventing dehydration. Carbohydrate contained in honey is believed can be served as an alternative form of carbohydrate for enhancing sports performance. Objective: To investigate the effectiveness of honey drink compared to sports drink as a recovery aid for running performance and physiological parameters in the heat. Method: Ten male recreational athletes (age: 22.2 ± 2.0 years, VO2max: 51.5 ± 3.7 ml.kg-1.min-1) participated in this randomized cross-over study. On each trial, participants were required to run for 1 hour in the glycogen depletion phase (Run-1), followed by a rehydration phase for 2 hours and subsequently a 20 minutes time trial performance (Run-2). During Run-1, subjects were required to run on the treadmill in the heat (31°C) with 70% relative humidity at 70 % of their VO2max. During rehydration phase, participants drank either honey drink or sports drink, or plain water with amount equivalent to 150% of body weight loss in dispersed interval (60 %, 50 % and 40 %) at 0 min, 30 min and 60 min respectively. Subsequently, time trial was performed by the participants in 20 minutes and the longest distance covered was recorded. Physiological parameters were analysed using two-way ANOVA with repeated measure and time trial performance was analysed using one-way ANOVA. Results: Result showed that Acacia honey elicited a better time trial performance with significantly longer distance compared to water trial (P<0.05). However, there was no significant difference between Acacia honey and sport drink trials (P > 0.05). Acacia honey and sports drink trials elicited 249 m (8.24 %) and 211 m (6.79 %) longer in distance compared to the water trial respectively. For physiological parameters, plasma glucose, plasma insulin and plasma free fatty acids in Acacia honey and sports drink trials were significantly higher compared to the water trial respectively during rehydration phase and time trial running performance phase. There were no significant differences in body weight changes, oxygen uptake, hematocrit, plasma volume changes and plasma cortisol in all the trials. Conclusion: Acacia honey elicited greatest beneficial effects on sports performance among the drinks, thus it has potential to be used for rehydration in athletes who train and compete in hot environment.

Keywords: honey drink, rehydration, sports performance, plasma glucose, plasma insulin, plasma cortisol

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Authors: Chee Wee Gan, Anthony Herr Cheun Ng, Yee Shan Wong, Subbu Venkatraman, Lynne Hsueh Yee Lim

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Otitis media with effusion (OME) is the leading cause of hearing loss in children worldwide. Surgery to insert grommet tube into the eardrum is usually indicated for OME unresponsive to antimicrobial therapy. It is the most common surgery for children. However, current commercially available grommet tubes are non-bioresorbable, not drug-treated, with unpredictable duration of retention on the eardrum to ventilate middle ear. Their functionality is impaired when clogged or chronically infected, requiring additional surgery to remove/reinsert grommet tubes. We envisaged that a novel fully bioresorbable grommet tube with sustained antibiotic release technology could address these drawbacks. In this study, drug-loaded bioresorbable poly(L-lactide-co-ε-caprolactone)(PLC) copolymer grommet tubes were fabricated by microinjection moulding technique. In vitro drug release and degradation model of PLC tubes were studied. Antibacterial property was evaluated by incubating PLC tubes with P. aeruginosa broth. Surface morphology was analyzed using scanning electron microscopy. A preliminary animal study was conducted using guinea pigs as an in vivo model to evaluate PLC tubes with and without drug, with commercial Mini Shah grommet tube as comparison. Our in vitro data showed sustained drug release over 3 months. All PLC tubes revealed exponential degradation profiles over time. Modeling predicted loss of tube functionality in water to be approximately 14 weeks and 17 weeks for PLC with and without drug, respectively. Generally, PLC tubes had less bacteria adherence, which were attributed to the much smoother tube surfaces compared to Mini Shah. Antibiotic from PLC tube further made bacteria adherence on surface negligible. They showed neither inflammation nor otorrhea after 18 weeks post-insertion in the eardrums of guinea pigs, but had demonstrated severe degree of bioresorption. Histology confirmed the new PLC tubes were biocompatible. Analyses on the PLC tubes in the eardrums showed bioresorption profiles close to our in vitro degradation models. The bioresorbable antibiotic-loaded grommet tubes showed good predictability in functionality. The smooth surface and sustained release technology reduced the risk of tube infection. Tube functional duration of 18 weeks allowed sufficient ventilation period to treat OME. Our ongoing studies include modifying the surface properties with protein coating, optimizing the drug dosage in the tubes to enhance their performances, evaluating their functional outcome on hearing after full resoption of grommet tube and healing of eardrums, and developing animal model with OME to further validate our in vitro models.

Keywords: bioresorbable polymer, drug release, grommet tube, guinea pigs, otitis media with effusion

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Authors: Chee Wai Ku, Pui See Chin

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As an emerging alternative to hysterectomy, uterine artery embolization (UAE) has been widely used in the management of fibroids and in controlling postpartum hemorrhage (PPH) unresponsive to other therapies. Research has shown UAE to be a safe, minimally invasive procedure with few complications and minimal effects on future fertility. We present two cases highlighting the use of UAE in preventing PPH in a patient with a large fibroid at the time of cesarean section and in the treatment of secondary PPH refractory to other therapies in another patient. We present a 36-year primiparous woman who booked at 18+6 weeks gestation with a 13.7 cm subserosal fibroid at the lower anterior wall of the uterus near the cervix and a 10.8 cm subserosal fibroid in the left wall. Prophylactic internal iliac artery occlusion balloons were placed prior to the planned classical midline cesarean section. The balloons were inflated once the baby was delivered. Bilateral uterine arteries were embolized subsequently. The estimated blood loss (EBL) was 400 mls and hemoglobin (Hb) remained stable at 10 g/DL. Ultrasound scan 2 years postnatally showed stable uterine fibroids 10.4 and 7.1 cm, which was significantly smaller than before. We present the second case of a 40-year-old G2P1 with a previous cesarean section for failure to progress. There were no antenatal problems, and the placenta was not previa. She presented with term labour and underwent an emergency cesarean section for failed vaginal birth after cesarean. Intraoperatively extensive adhesions were noted with bladder drawn high, and EBL was 300 mls. Postpartum recovery was uneventful. She presented with secondary PPH 3 weeks later complicated by hypovolemic shock. She underwent an emergency examination under anesthesia and evacuation of the uterus, with EBL 2500mls. Histology showed decidua with chronic inflammation. She was discharged well with no further PPH. She subsequently returned one week later for secondary PPH. Bedside ultrasound showed that the endometrium was thin with no evidence of retained products of conception. Uterotonics were administered, and examination under anesthesia was performed, with uterine Bakri balloon and vaginal pack insertion after. EBL was 1000 mls. There was no definite cause of PPH with no uterine atony or products of conception. To evaluate a potential cause, pelvic angiogram and super selective left uterine arteriogram was performed which showed profuse contrast extravasation and acute bleeding from the left uterine artery. Superselective embolization of the left uterine artery was performed. No gross contrast extravasation from the right uterine artery was seen. These two cases demonstrated the superior efficacy of UAE. Firstly, the prophylactic use of intra-arterial balloon catheters in pregnant patients with large fibroids, and secondly, in the diagnosis and management of secondary PPH refractory to uterotonics and uterine tamponade. In both cases, the need for laparotomy hysterectomy was avoided, resulting in the preservation of future fertility. UAE should be a consideration for hemodynamically stable patients in centres with access to interventional radiology.

Keywords: fertility preservation, secondary postpartum hemorrhage, uterine embolization, uterine fibroids

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Authors: Yi H. Wong, Li L. Chan, Chee O. Leong, Stephen Ambu, Joon W. Mak, Priyadashi S. Sahu

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Background: Acanthamoeba is a free-living opportunistic protist which is ubiquitously distributed in the environment. Virulent Acanthamoeba can cause fatal encephalitis in immunocompromised patients and potential blinding keratitis in immunocompetent contact lens wearers. Approximately 24 species have been identified but only the A. castellanii, A. polyphaga and A. culbertsoni are commonly associated with human infections. Until to date, the precise molecular basis for Acanthamoeba pathogenesis remains unclear. Previous studies reported that Acanthamoeba virulence can be diminished through prolonged axenic culture but revived through serial mouse passages. As no clear explanation on this reversible pathogenesis is established, hereby, we postulate that the epigenetic regulators, DNA-methyltransferases (DNMT) and histone-deacetylases (HDAC), could possibly be involved in granting the virulence plasticity of Acanthamoeba spp. Methods: Four rounds of mouse passages were conducted to revive the virulence potential of the virulence-attenuated Acanthamoeba castellanii strain (ATCC 50492). Briefly, each mouse (n=6/group) was inoculated intraperitoneally with Acanthamoebae cells (2x 105 trophozoites/mouse) and incubated for 2 months. Acanthamoebae cells were isolated from infected mouse organs by culture method and subjected to subsequent mouse passage. In vitro cytopathic, encystment and gelatinolytic assays were conducted to evaluate the virulence characteristics of Acanthamoebae isolates for each passage. PCR primers which targeted on the 2 members (DNMT1 and DNMT2) and 5 members (HDAC1 to 5) of the DNMT and HDAC gene families respectively were custom designed. Quantitative real-time PCR (qPCR) was performed to detect and quantify the relative expression of the two gene families in each Acanthamoeba isolates. Beta-tubulin of A. castellanii (Genbank accession no: XP_004353728) was included as housekeeping gene for data normalisation. PCR mixtures were also analyzed by electrophoresis for amplicons detection. All statistical analyses were performed using the paired one-tailed Student’s t test. Results: Our pathogenicity tests showed that the virulence-reactivated Acanthamoeba had a higher degree of cytopathic effect on vero cells, a better resistance to encystment challenge and a higher gelatinolytic activity which was catalysed by serine protease. qPCR assay showed that DNMT1 expression was significantly higher in the virulence-reactivated compared to the virulence-attenuated Acanthamoeba strain (p ≤ 0.01). The specificity of primers which targeted on DNMT1 was confirmed by sequence analysis of PCR amplicons, which showed a 97% similarity to the published DNA-methyltransferase gene of A. castellanii (GenBank accession no: XM_004332804.1). Out of the five primer pairs which targeted on the HDAC family genes, only HDAC4 expression was significantly difference between the two variant strains. In contrast to DNMT1, HDAC4 expression was much higher in the virulence-attenuated Acanthamoeba strain. Conclusion: Our mouse passages had successfully restored the virulence of the attenuated strain. Our findings suggested that DNA-methyltransferase (DNMT1) and histone deacetylase (HDAC4) expressions are associated with virulence potential of Acanthamoeba spp.

Keywords: acanthamoeba, DNA-methyltransferase, histone deacetylase, virulence-associated proteins

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Authors: Yi-Hao Wong, Li-Li Chan, Chee-Onn Leong, Stephen Ambu, Joon-Wah Mak, Priyasashi Sahu

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Background: Acanthamoeba is a genus of amoebae which lives as a free-living in nature or as a human pathogen that causes severe brain and eye infections. Virulence potential of Acanthamoeba is not constant and can change with growth conditions. DNA methylation, an epigenetic process which adds methyl groups to DNA, is used by eukaryotic cells, including several human parasites to control their gene expression. We used qPCR, siRNA gene silencing, and RNA sequencing (RNA-Seq) to study DNA-methyltransferase gene family (DNMT) in order to indicate the possibility of its involvement in programming Acanthamoeba virulence potential. Methods: A virulence-attenuated Acanthamoeba isolate (designation: ATCC; original isolate: ATCC 50492) was subjected to mouse passages to restore its pathogenicity; a virulence-reactivated isolate (designation: AC/5) was generated. Several established factors associated with Acanthamoeba virulence phenotype were examined to confirm the succession of reactivation process. Differential gene expression of DNMT between ATCC and AC/5 isolates was performed by qPCR. Silencing on DNMT gene expression in AC/5 isolate was achieved by siRNA duplex. Total RNAs extracted from ATCC, AC/5, and siRNA-treated (designation: si-146) were subjected to RNA-Seq for comparative transcriptomic analysis in order to identify the genome-wide effect of DNMT in regulating Acanthamoeba gene expression. qPCR was performed to validate the RNA-Seq results. Results: Physiological and cytophatic assays demonstrated an increased in virulence potential of AC/5 isolate after mouse passages. DNMT gene expression was significantly higher in AC/5 compared to ATCC isolate (p ≤ 0.01) by qPCR. si-146 duplex reduced DNMT gene expression in AC/5 isolate by 30%. Comparative transcriptome analysis identified the differentially expressed genes, with 3768 genes in AC/5 vs ATCC isolate; 2102 genes in si-146 vs AC/5 isolate and 3422 genes in si-146 vs ATCC isolate, respectively (fold-change of ≥ 2 or ≤ 0.5, p-value adjusted (padj) < 0.05). Of these, 840 and 1262 genes were upregulated and downregulated, respectively, in si-146 vs AC/5 isolate. Eukaryotic orthologous group (KOG) assignments revealed a higher percentage of downregulated gene expression in si-146 compared to AC/5 isolate, were related to posttranslational modification, signal transduction and energy production. Gene Ontology (GO) terms for those downregulated genes shown were associated with transport activity, oxidation-reduction process, and metabolic process. Among these downregulated genes were putative genes encoded for heat shock proteins, transporters, ubiquitin-related proteins, proteins for vesicular trafficking (small GTPases), and oxidoreductases. Functional analysis of similar predicted proteins had been described in other parasitic protozoa for their survival and pathogenicity. Decreased expression of these genes in si146-treated isolate may account in part for Acanthamoeba reduced pathogenicity. qPCR on 6 selected genes upregulated in AC/5 compared to ATCC isolate corroborated the RNA sequencing findings, indicating a good concordance between these two analyses. Conclusion: To the best of our knowledge, this study represents the first genome-wide analysis of DNA methylation and its effects on gene expression in Acanthamoeba spp. The present data indicate that DNA methylation has substantial effect on global gene expression, allowing further dissection of the genome-wide effects of DNA-methyltransferase gene in regulating Acanthamoeba pathogenicity.

Keywords: Acanthamoeba, DNA methylation, RNA sequencing, virulence

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Authors: Mei Xuan Tan, Gim-Yang Maggie Pee, Mei Chee Tan

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Academic performance, in the form of scoring high grades in enrolled subjects, is not the only significant trait in achieving success. Engineering graduates with experience in working on hands-on projects in a team setting are highly sought after in industry upon graduation. Such projects are typically real world problems that require the integration and application of knowledge and skills from several disciplines. In a traditional university setting, subjects are taught in a silo manner with no cross participation from other departments or disciplines. This may lead to knowledge compartmentalization and students are unable to understand and connect the relevance and applicability of the subject. University instructors thus see this integration across disciplines as a challenging task as they aim to better prepare students in understanding and solving problems for work or future studies. To improve students’ academic performance and to cultivate various skills such as critical thinking, there has been a gradual uptake in the use of an active learning approach in introductory science and engineering courses, where lecturing is traditionally the main mode of instruction. This study aims to discuss the implementation and experience of a hands-on, interdisciplinary project that involves all the four core subjects taught during the term at the Singapore University of Technology Design (SUTD). At SUTD, an interdisciplinary design activity, named 2D, is integrated into the curriculum to help students reinforce the concepts learnt. A student enrolled in SUTD experiences his or her first 2D in Term 1. This activity. which spans over one week in Week 10 of Term 1, highlights the application of chemistry, physics, mathematics, humanities, arts and social sciences (HASS) in designing an engineering product solution. The activity theme for Term 1 2D revolved around “work and play”. Students, in teams of 4 or 5, used a scaled-down model of a chemical launcher to launch a projectile across the room. It involved the use of a small chemical combustion reaction between ethanol (a highly volatile fuel) and oxygen. This reaction generated a sudden and large increase in gas pressure built up in a closed chamber, resulting in rapid gas expansion and ejection of the projectile out of the launcher. Students discussed and explored the meaning of play in their lives in HASS class while the engineering aspects of a combustion system to launch an object using underlying principles of energy conversion and projectile motion were revisited during the chemistry and physics classes, respectively. Numerical solutions on the distance travelled by the projectile launched by the chemical launcher, taking into account drag forces, was developed during the mathematics classes. At the end of the activity, students developed skills in report writing, data collection and analysis. Specific to this 2D activity, students gained an understanding and appreciation on the application and interdisciplinary nature of science, engineering and HASS. More importantly, students were exposed to design and problem solving, where human interaction and discussion are important yet challenging in a team setting.

Keywords: active learning, collaborative learning, first year undergraduate, interdisciplinary, STEAM

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