Search results for: D. T. Leong

Authors: Rozita Borhan, Hanif Che Lah, Wee Leong Son

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This paper is about method to produce a stable and accurate constant output pulse width regardless of the amplitude, period and pulse width variation of the input signal source. The pulse generated is usually being used in numerous applications as the reference input source to other circuits in the system. Therefore, it is crucial to produce a clean and constant pulse width to make sure the system is working accurately as expected.

Keywords: amplitude, Constant Pulse Width, frequency divider, pulse generator

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Authors: Fakhreddin Abedi, Wah June Leong

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Exponential stability of stochastic differential equations with non trivial solutions is provided in terms of Lyapunov functions. The main result of this paper establishes that, under certain hypotheses for the dynamics f(.) and g(.), practical exponential stability in probability at the small neighborhood of the origin is equivalent to the existence of an appropriate Lyapunov function. Indeed, we establish exponential stability of stochastic differential equation when almost all the state trajectories are bounded and approach a sufficiently small neighborhood of the origin. We derive sufficient conditions for exponential stability of stochastic differential equations. Finally, we give a numerical example illustrating our results.

Keywords: exponential stability in probability, stochastic differential equations, Lyapunov technique, Ito's formula

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Authors: Cedric Leong, Parth Desai, Parth Patel

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The purpose of this project was to develop a neural network based on qualitative team data to predict alliance scores to determine winners of matches in the FIRST Robotics Competition (FRC). The game for the competition changes every year with different objectives and game objects, however the idea was to create a prediction system which can be reused year by year using some of the statistics that are constant through different games, making our system adaptable to future games as well. Aerial Assist is the FRC game for 2014, and is played in alliances of 3 teams going against one another, namely the Red and Blue alliances. This application takes any 6 teams paired into 2 alliances of 3 teams and generates the prediction for the final score between them.

Keywords: artifical neural network, prediction system, qualitative team data, FIRST Robotics Competition (FRC)

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Authors: Leong Tatt Loh, Ming Kun Yew, Ming Chian Yew, Lip Huat Saw, Jing Han Beh, Siong Kang Lim, Foo Wei Lee

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Oil palm shell (OPS) is the solid waste product from the palm oil sector of the agricultural industry and can be used as alternative coarse aggregates to substitute depleting conventional raw materials. This research aims to investigate the incorporation of various high-performance polypropylene (HPP) fibres with different geometry to enhance the mechanical properties and thermal conductivity of OPS lightweight concrete. The effect of different volume fractions (Vf) (0.05%, 0.10% and 0.15%) were studied for each fibre. The results reveal that the effectiveness of HPP fibres to increase the compressive strength at later ages was more pronounced than at early age. It is found that the use of HPP fibres reinforced OPS lightweight concrete (LWC) induced the advantageous of improving mechanical properties (compressive strength, flexural strength and splitting tensile strength) and thermal conductivity. Hence, this HPP fibres is a promising alternative solution to compensate lower mechanical properties as well as contribute to energy efficiency building material in the construction industry.

Keywords: oil palm shell, high performance polypropylene fibre, lightweight concrete, mechanical properties, thermal conductivity

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Authors: Sie Long Kek, Wah June Leong, Kok Lay Teo

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Linear quadratic Gaussian model is a standard mathematical model for the stochastic optimal control problem. The combination of the linear quadratic estimation and the linear quadratic regulator allows the state estimation and the optimal control policy to be designed separately. This is known as the separation principle. In this paper, an efficient computational method is proposed to solve the linear quadratic Gaussian problem. In our approach, the Hamiltonian function is defined, and the necessary conditions are derived. In addition to this, the output error is defined and the least-square optimization problem is introduced. By determining the first-order necessary condition, the gradient of the sum squares of output error is established. On this point of view, the stochastic approximation approach is employed such that the optimal control policy is updated. Within a given tolerance, the iteration procedure would be stopped and the optimal solution of the linear-quadratic Gaussian problem is obtained. For illustration, an example of the linear-quadratic Gaussian problem is studied. The result shows the efficiency of the approach proposed. In conclusion, the applicability of the approach proposed for solving the linear quadratic Gaussian problem is highly demonstrated.

Keywords: iteration procedure, least squares solution, linear quadratic Gaussian, output error, stochastic approximation

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Authors: Adam Kok Wey Leong

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Special operations were supposedly a new way of irregular warfare that was officially formed during World War 2. For example, the famous British Special Operations Executive (SOE) and the Americans’ Office for Strategic Services (OSS) – the forerunners of modern day CIA were born in World War 2. These special operations units were tasked with the conduct of sabotage and subversion activities behind enemy lines, placing great importance in forming Fifth Column activities and supporting resistance movements. This pointed to a paradoxical argument that modern day special operations is a product of Western modern military innovation but utilizing Eastern ways of ‘ungentlemanly’ warfare. This thesis is superfluous as special operations had been well practised by both ancient Western empires such as the Greeks and Romans, and around the same time in the East, such as in China, and Japan. This paper will describe the practice of special operations, first from the Western military history of the Greeks during the Peloponnesian war. It will then highlight the similar practice of special operations by the Near Eastern Assassins and Eastern militaries by using examples from the Chinese and the Japanese. This paper propounds that special operations, or ways of warfare as a whole, has no cultural and geographical divide, but rather very similarly practiced by men from all over the world. Ideas of fighting, killing and ultimately winning a war have similar undertones – attempts to find ways to win economically and at the least time.

Keywords: special operations, strategic culture, ways of warfare, Sun Tzu, Frontinus

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Authors: Adam Kok Wey Leong

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The comparison between the Western ways of war and Eastern ways of war has raised contemporary debates on the validity of these arguments. The Western way of war is popularly propounded by Victor Davis Hanson as originating from the Greek hoplite tactics, direct military maneuvers, democratic principles and social freedom and cohesion that has continued to yield military success for the Western powers for centuries. On the other hand, the Eastern way of war has been deemed as relying on indirect tactics, deception, and ruses. This often accepted notion of the divide between Western and Eastern style does not sustain in view of the available classical strategic texts from both sides from the same period that has proposed similar principles of warfare. This paper analyses the similarities between classical strategic texts on war from the Eastern perspective namely Sun Tzu’s Art of War with a similar temporal strategic text from the West which is Sextus Iuluis Frontinus’s Stratagematon, and deduces answers to this core research question - Does the hypothesis of the existence of distinctive Western and Eastern ways of warfare stands? The main thesis advanced by this research is that ways of warfare share universal principles, and it transcends cultural and spatial boundaries. Warfare is a human endeavour, and the same moral actions guide humans from different geo-cultural spheres in warfare’s objectives, which are winning over an enemy in the most economical way and serve as a mean to an end.

Keywords: ways of warfare, strategic culture, strategy, Sun Tzu, frontinus

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Authors: Y. K. Leong, J. C. W. Lan, H. S. Loh, P. L. Show

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Being biodegradable, non-toxic, renewable and have similar or better properties as commercial plastics, polyhydroxy alkanoates (PHAs) can be a potential game changer in the polymer industry. PHAs are the biodegradable polymer produced by bacteria, which are in interest as a sustainable alternative to petrochemical-derived plastics; however, its commercial value has significantly limited by high production and recovery cost of PHA. Aqueous two-phase system (ATPS) offers different chemical and physical environments, which contains about 80-90% water delivers an excellent environment for partitioning of cells, cell organelles and biologically active substances. Extractive bioconversion via ATPS allows the integration of PHA upstream fermentation and downstream purification process, which reduces production steps and time, thus lead to cost reduction. The ability of Ralstonia eutropha to grow under different ATPS conditions was investigated for its potential to be used in a bioconversion system. Changes in tie-line length (TLL) and a volume ratio (Vr) were shown to have an effect on PHA partition coefficient. High PHA recovery yield of 65% with a relatively high purity of 73% was obtained in PEG 6000/Sodium sulphate system with 42.6 wt/wt % TLL and 1.25 Vr. Extractive bioconversion via ATPS is an attractive approach for the combination of PHA production and recovery process.

Keywords: aqueous two-phase system, extractive bioconversion, polyhydroxy alkanoates, purification

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Authors: Aliana Leong, T. C. Huan

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The purpose of Nostalgic themed tourism product is becoming popular in many countries. This study intends to investigate the role of nostalgia in destination image, experiential value and their effect on subsequent behavioral intention. The survey used stratified sampling method to include respondents from all the nearby Asian regions. The sampling is based on the data of inbound tourists provided by the Statistics and Census Service (DSEC) of government of Macau. The questionnaire consisted of five sections of 5 point Likert scale questions: (1) nostalgia, (2) destination image both before and after experience, (3) expected value, (4) experiential value, and (5) future visit intention. Data was analysed with structural equation modelling. The result indicates that nostalgia plays an important part in forming destination image and experiential value before individual had a chance to experience the destination. The destination image and experiential value share a bidirectional causal relationship that eventually contributes to future visit intention. The study also discovered that while experiential value is more effective in generating destination image, the later contribute more to future visit intention. The research design measures destination image and experiential value before and after respondents had experience the destination. The distinction between destination image and expected/experiential value can be examined because the longitudinal design of research method. It also allows this study to observe how nostalgia translates to future visit intention.

Keywords: nostalgia, destination image, experiential value, future visit intention

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Authors: Yin-Yee Leong, Jack C. Yue, Hsin-Chung Wang

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Human longevity has been experiencing its largest increase since the end of World War II, and modeling the mortality rates is therefore often the focus of many studies. Among all mortality models, the Lee–Carter model is the most popular approach since it is fairly easy to use and has good accuracy in predicting mortality rates (e.g., for Japan and the USA). However, empirical studies from several countries have shown that the age parameters of the Lee–Carter model are not constant in time. Many modifications of the Lee–Carter model have been proposed to deal with this problem, including adding an extra cohort effect and adding another period effect. In this study, we propose a spatial modification and use clusters to explain why the age parameters of the Lee–Carter model are not constant. In spatial analysis, clusters are areas with unusually high or low mortality rates than their neighbors, where the “location” of mortality rates is measured by age and time, that is, a 2-dimensional coordinate. We use a popular cluster detection method—Spatial scan statistics, a local statistical test based on the likelihood ratio test to evaluate where there are locations with mortality rates that cannot be described well by the Lee–Carter model. We first use computer simulation to demonstrate that the cluster effect is a possible source causing the problem of the age parameters not being constant. Next, we show that adding the cluster effect can solve the non-constant problem. We also apply the proposed approach to mortality data from Japan, France, the USA, and Taiwan. The empirical results show that our approach has better-fitting results and smaller mean absolute percentage errors than the Lee–Carter model.

Keywords: mortality improvement, Lee–Carter model, spatial statistics, cluster detection

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Authors: H. Z. M. Chong, M. E. Y. Leong, G. L. Khor, S. C. Loke

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Vitamin D insufficiency is reported to be prevalent among women living in different altitudes including the equator where sunshine is available throughout the year. Multiple factors for vitamin D insufficiency include poor intake of vitamin D rich food and inadequate sun exposure, especially among women working indoor with a sedentary lifestyle. Furthermore, Muslim women in Malaysia whose attire covers the entire body are likely to receive poor sun exposure. This research determined serum vitamin D status, vitamin D intake and sun exposure of women aged 20-45 years of different ethnicity in Kuala Lumpur, Malaysia. Blood samples were collected from 106 women for determination of serum 25(OH)D levels. Information about vitamin D intake and sun exposure were obtained by interviewing the subjects using pre-tested questionnaires. The overall mean serum 25(OH)D was found to be 29.9 ± 14 nmol/L. Vitamin D deficiency and insufficiency was prevalent and highest among the Malay women. Less than ten percent of the subjects in this study met the sufficient vitamin D level recommendation of ≥50 nmol/L. Intake of vitamin D rich food such as oily fishes was poor across the different ethnicity. Other dietary sources of vitamin D in the diet were fortified bread and skim milk. On the other hand, the median sunlight exposure of the subjects was 3.9 hours per week. The Malay women reported to have the highest duration being exposed to the sun. Nevertheless, due to cultural clothing practices, these women had the least body surface area exposed to sunlight, resulting in the lowest calculated sun index score compared to the Chinese and the Indians. Low intake of vitamin D rich foods and sun exposure were negatively correlated with serum 25(OH)D level. In conclusion, intake of food sources rich in vitamin D and adequate body surface area exposed to the sun are essential to ensure healthy vitamin D level. Supplementation of vitamin D may be recommended to women whom unable to meet these recommendations.

Keywords: serum 25-OH, sun exposure, vitamin D food frequency, vitamin D deficiency

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Authors: Narash Narasimman, Calvin Leong Jia Jun, Raksha Karthik, Paul Englert

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Background: Project Grit, a 12-week youth resilience program implemented by Impart and Spartans Boxing Club, aimed to help at-risk adolescents develop resilience through psychoeducation and mental health techniques for dealing with everyday stressors and adversity. The programme consists of two parts-1.5 hours of group therapy followed by 1 hour of boxing. Due to the novelty of the study, 6 male participants, aged 13 to 18, were recruited to participate in the study. Aim: This case study aims to examine the effectiveness of Project Grit in building resilience among at-risk adolescents. Methods: A case study design was employed to capture the complexity and uniqueness of the intervention, without oversimplifying or generalizing it. A 15-year-old male participant with a history of behavioural challenges, delinquency and gang involvement was selected for the study. Teacher, parent and child versions of the Strengths and Difficulties Questionnaire (SDQ) were administered to the facilitators, parents and participants respectively before and after the programme. Relevant themes from the qualitative interviews will be discussed. Results: Scores from all raters revealed improvements in most domains of the SDQ. Total difficulties scores across all raters improved from “very high” to “close to average”. High interrater reliability was observed (κ= .81). The participant reported learning methods to effectively deal with his everyday concerns using healthy coping strategies, developing a supportive social network, and building on his self efficacy. Themes from the subject’s report concurred with the improvement in SDQ scores. Conclusions: The findings suggest that Project Grit is a promising intervention for promoting resilience among at-risk adolescents. The teleological behaviourism framework and the combination of sports engagement and future orientation may be particularly effective in fostering resilience among this population. Further studies need to be conducted with a larger sample size to further validate the effectiveness of Project Grit.

Keywords: resilience, project grit, adolescents, at-risk, boxing, future orientation

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Authors: May Khei Hu, Kevin Leong, Fu Siong Ng, Nicholas Peter

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Introduction: Sympathetic stimulation has been recognised as a potent stimulus of arrhythmogenesis in various cardiac pathologies, possibly by augmenting dispersion of repolarisation. The effects of sympathetic stimulation in healthy subjects however remain unclear. It is, therefore, crucial to first establish the effects of physiological stress on dispersion of repolarisation in healthy subjects before understanding these effects in pathological cardiac conditions. We hypothesised that activation-recovery interval (ARI; which is a surrogate of action potential duration) and dispersion of repolarisation decrease on sympathetic stimulation. Methods: Eight patients aged 18-55 years with structurally normal hearts underwent head-up tilt test (HUTT) and exercise tolerance test (ETT) while wearing the electrocardiographic imaging (ECGi) vest. Patients later underwent CT scan and the epicardial potentials are reconstructed using the ECGi software. Activation and recovery times were determined from the acquired electrograms. ARI was calculated and later corrected using Bazett’s formula. Global and regional dispersion of repolarisation were determined from standard deviation of the corrected ARI (ARIc). One-way analysis of variance (ANOVA) and Wilcoxon test were used to evaluate statistical significance. Results: Global ARIc increased significantly [p<0.01] when patients were tilted upwards but decreased significantly after five minutes [p<0.01]. A subsequent post- hoc analysis revealed that the decrease in R-R was more substantial compared to the change in ARI, resulting in the observed increase in ARIc. Global ARIc decreased on peak exercise [p<0.01] but increased on recovery [p<0.01]. Global dispersion increased significantly on peak exercise [p<0.05] although there were no significant changes in regional dispersion. There were no significant changes in both global and regional dispersion during tilt. Conclusion: ARIc decreases upon sympathetic stimulation in healthy subjects. Global dispersion of repolarisation increases upon exercise although there were no changes in global or regional dispersion during orthostatic stress.

Keywords: dispersion of repolarisation, sympathetic stimulation, Head-up tilt test (HUTT), Exercise tolerance test (ETT), Electrocardiographic imaging (ECGi)

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Authors: Eugene Y. J. Aw, J. W. Koh, S. H. Chew, K. E. Chua, Lionel L. J. Ang, Algernon C. S. Hong, Danette S. E. Tan, Grace H. B. Foo, K. Q. Hong, L. M. Cheng, M. L. Leong

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This paper presents a novel rapid soil classification technique that combines computer vision with four-probe soil electrical resistivity method and cone penetration test (CPT), to improve the accuracy and productivity of on-site classification of excavated soil. In Singapore, excavated soils from local construction projects are transported to Staging Grounds (SGs) to be reused as fill material for land reclamation. Excavated soils are mainly categorized into two groups (“Good Earth” and “Soft Clay”) based on particle size distribution (PSD) and water content (w) from soil investigation reports and on-site visual survey, such that proper treatment and usage can be exercised. However, this process is time-consuming and labour-intensive. Thus, a rapid classification method is needed at the SGs. Computer vision, four-probe soil electrical resistivity and CPT were combined into an innovative non-destructive and instantaneous classification method for this purpose. The computer vision technique comprises soil image acquisition using industrial grade camera; image processing and analysis via calculation of Grey Level Co-occurrence Matrix (GLCM) textural parameters; and decision-making using an Artificial Neural Network (ANN). Complementing the computer vision technique, the apparent electrical resistivity of soil (ρ) is measured using a set of four probes arranged in Wenner’s array. It was found from the previous study that the ANN model coupled with ρ can classify soils into “Good Earth” and “Soft Clay” in less than a minute, with an accuracy of 85% based on selected representative soil images. To further improve the technique, the soil strength is measured using a modified mini cone penetrometer, and w is measured using a set of time-domain reflectometry (TDR) probes. Laboratory proof-of-concept was conducted through a series of seven tests with three types of soils – “Good Earth”, “Soft Clay” and an even mix of the two. Validation was performed against the PSD and w of each soil type obtained from conventional laboratory tests. The results show that ρ, w and CPT measurements can be collectively analyzed to classify soils into “Good Earth” or “Soft Clay”. It is also found that these parameters can be integrated with the computer vision technique on-site to complete the rapid soil classification in less than three minutes.

Keywords: Computer vision technique, cone penetration test, electrical resistivity, rapid and non-destructive, soil classification

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Authors: Eugene Y. J. Aw, J. W. Koh, S. H. Chew, K. E. Chua, P. L. Goh, Grace H. B. Foo, M. L. Leong

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This paper presents the evaluation of various soil testing methods such as the four-probe soil electrical resistivity method and cone penetration test (CPT) that can complement a newly developed novel rapid soil classification scheme using computer vision, to improve the accuracy and productivity of on-site classification of excavated soil. In Singapore, excavated soils from the local construction industry are transported to Staging Grounds (SGs) to be reused as fill material for land reclamation. Excavated soils are mainly categorized into two groups (“Good Earth” and “Soft Clay”) based on particle size distribution (PSD) and water content (w) from soil investigation reports and on-site visual survey, such that proper treatment and usage can be exercised. However, this process is time-consuming and labor-intensive. Thus, a rapid classification method is needed at the SGs. Four-probe soil electrical resistivity and CPT were evaluated for their feasibility as suitable additions to the computer vision system to further develop this innovative non-destructive and instantaneous classification method. The computer vision technique comprises soil image acquisition using an industrial-grade camera; image processing and analysis via calculation of Grey Level Co-occurrence Matrix (GLCM) textural parameters; and decision-making using an Artificial Neural Network (ANN). It was found from the previous study that the ANN model coupled with ρ can classify soils into “Good Earth” and “Soft Clay” in less than a minute, with an accuracy of 85% based on selected representative soil images. To further improve the technique, the following three items were targeted to be added onto the computer vision scheme: the apparent electrical resistivity of soil (ρ) measured using a set of four probes arranged in Wenner’s array, the soil strength measured using a modified mini cone penetrometer, and w measured using a set of time-domain reflectometry (TDR) probes. Laboratory proof-of-concept was conducted through a series of seven tests with three types of soils – “Good Earth”, “Soft Clay,” and a mix of the two. Validation was performed against the PSD and w of each soil type obtained from conventional laboratory tests. The results show that ρ, w and CPT measurements can be collectively analyzed to classify soils into “Good Earth” or “Soft Clay” and are feasible as complementing methods to the computer vision system.

Keywords: computer vision technique, cone penetration test, electrical resistivity, rapid and non-destructive, soil classification

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Authors: V. K. Rajaletchumy, S. L. Chia, M. I. Setyawati, M. S. Muthu, S. S. Feng, D. T. Leong

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Triple negative breast cancers (TNBC) can be classified as one of the most aggressive with a high rate of local recurrences and systematic metastases. TNBCs are insensitive to existing hormonal therapy or targeted therapies such as the use of monoclonal antibodies, due to the lack of oestrogen receptor (ER) and progesterone receptor (PR) and the absence of overexpression of human epidermal growth factor receptor 2 (HER2) compared with other types of breast cancers. The absence of targeted therapies for selective delivery of therapeutic agents into tumours, led to the search for druggable targets in TNBC. In this study, we developed a targeted micellar system of cetuximab-conjugated micelles of D-α-tocopheryl polyethylene glycol succinate (vitamin E TPGS) for targeted delivery of docetaxel as a model anticancer drug for the treatment of TNBCs. We examined the efficacy of our micellar system in xenograft models of triple negative breast cancers and explored the effect of the micelles on post-treatment tumours in order to elucidate the mechanism underlying the nanomedicine treatment in oncology. The targeting micelles were found preferentially accumulated in tumours immediately after the administration of the micelles compare to normal tissue. The fluorescence signal gradually increased up to 12 h at the tumour site and sustained for up to 24 h, reflecting the increases in targeted micelles (TPFC) micelles in MDA-MB-231/Luc cells. In comparison, for the non-targeting micelles (TPF), the fluorescence signal was evenly distributed all over the body of the mice. Only a slight increase in fluorescence at the chest area was observed after 24 h post-injection, reflecting the moderate uptake of micelles by the tumour. The successful delivery of docetaxel into tumour by the targeted micelles (TPDC) exhibited a greater degree of tumour growth inhibition than Taxotere® after 15 days of treatment. The ex vivo study has demonstrated that tumours treated with targeting micelles exhibit enhanced cell cycle arrest and attenuated proliferation compared with the control and with those treated non-targeting micelles. Furthermore, the ex vivo investigation revealed that both the targeting and non-targeting micellar formulations shows significant inhibition of cell migration with migration indices reduced by 0.098- and 0.28-fold, respectively, relative to the control. Overall, both the in vivo and ex vivo data increased the confidence that our micellar formulations effectively targeted and inhibited EGF-overexpressing MDA-MB-231 tumours.

Keywords: biodegradable polymers, cancer nanotechnology, drug targeting, molecular biomaterials, nanomedicine

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Authors: Antonio Ruiz Gonzalez, Kwang-Leong Choy

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The measurement of electrolytes has a high value in the clinical routine. Ions are present in all body fluids with variable concentrations and are involved in multiple pathologies such as heart failures and chronic kidney disease. In the case of dissolved potassium, although a high concentration in the blood (hyperkalemia) is relatively uncommon in the general population, it is one of the most frequent acute electrolyte abnormalities. In recent years, the integration of thin films technologies in this field has allowed the development of highly sensitive biosensors with ultra-low limits of detection for the assessment of metals in liquid samples. However, despite the current efforts in the miniaturization of sensitive devices and their integration into portable systems, only a limited number of successful examples used commercially can be found. This fact can be attributed to a high cost involved in their production and the sustained degradation of the electrodes over time, which causes a signal drift in the measurements. Thus, there is an unmet necessity for the development of low-cost and robust sensors for the real-time monitoring of analyte concentrations in patients to allow the early detection and diagnosis of diseases. This paper reports a thin film ion-selective sensor for the evaluation of potassium ions in aqueous samples. As an alternative for this fabrication method, aerosol assisted chemical vapor deposition (AACVD), was applied due to cost-effectivity and fine control over the film deposition. Such a technique does not require vacuum and is suitable for the coating of large surface areas and structures with complex geometries. This approach allowed the fabrication of highly homogeneous surfaces with well-defined microstructures onto 50 nm thin gold layers. The degradative processes of the ubiquitously employed poly (vinyl chloride) membranes in contact with an electrolyte solution were studied, including the polymer leaching process, mechanical desorption of nanoparticles and chemical degradation over time. Rational design of a protective coating based on an organosilicon material in combination with cellulose to improve the long-term stability of the sensors was then carried out, showing an improvement in the performance after 5 weeks. The antifouling properties of such coating were assessed using a cutting-edge quartz microbalance sensor, allowing the quantification of the adsorbed proteins in the nanogram range. A correlation between the microstructural properties of the films with the surface energy and biomolecules adhesion was then found and used to optimize the protective film.

Keywords: hyperkalemia, drift, AACVD, organosilicon

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Authors: Shirley S. Ho, Alisius X. L. D. Leong, Jiemin Looi, Agnes S. F. Chuah

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Nuclear energy is a contentious technology that has attracted much public debate over the years. The prominence of nuclear energy in Southeast Asia (SEA) has burgeoned due to the surge of interest and plans for nuclear development in the region. Understanding public perceptions of nuclear energy in SEA is pertinent given the limited number of studies conducted. In particular, five SEA nations – Singapore, Malaysia, Indonesia, Thailand, and Vietnam are of immediate interest as that they are amongst the most economically developed or developing nations in the SEA region. High energy demands from economic development in these nations have led to considerations of adopting nuclear energy as an alternative source of energy. This study aims to explore whether differences in the nuclear developmental stage in each country affects public perceptions of nuclear energy. In addition, this study seeks to find out about the type and importance of communication credibility as a judgement heuristic in facilitating message acceptance across these five countries. Credibility of a communication channel is a crucial component influencing public perception, acceptance, and attitudes towards nuclear energy. Aside from simply identifying the frequently used communication channels, it is of greater significance to understand public perception of source and media credibility. Given the lack of studies conducted in SEA, this exploratory study adopts a qualitative approach to elicit a spectrum of opinions and insights regarding the key communication aspects influencing public perceptions of nuclear energy. Specifically, the capitals of each of the abovementioned countries - Kuala Lumpur, Bangkok, and Hanoi - were selected, with the exception of Singapore, an island city-state, and Yogyakarta, the most populous island of Indonesia to better understand public perception towards nuclear energy. Focus group discussions were utilized as the mode of data collection to elicit a wide variety of viewpoints held by the participants, which is well-suited for exploratory research. In total, 156 participants took part in the 13 focus group discussions. The participants were either local citizens or permanent residents aged between 18 and 69 years old. Each of the focus groups consists of 8-10 participants, including both male and female participants. The transcripts from each focus group were analysed using NVivo 10, and the text was organised according to the emerging themes or categories. The general public in all the countries was familiar but had no in-depth knowledge with nuclear energy. Four dimensions of nuclear energy communication were identified based on the focus group discussions: communication channels, perceived credibility of sources, circumstances for discussion, and discussion style. The first dimension, communication channels refers to the medium through which participants receive information about nuclear energy. Four types of media emerged from the discussions. They included online and social media, broadcast media, print media, and word-of- mouth (WOM). Collectively, across all five countries, participants were found to engage in different types of knowledge acquisition and information seeking behavior depending on the communication channels used.

Keywords: nuclear energy, public perception, communication, Southeast Asia, source credibility

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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

Abstract:

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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