Construction and characterization of H5N1-recombinant fowlpox viruses co-expressing host cytokines

Possessing a large double stranded DNA genome up to 300 kb, fowlpox virus (FWPV) has been developed to express avian influenza virus (AIV) antigens since the late 1980s. A more advanced approach would be to coexpress host cytokines from such recombinants. This thesis describes the strategy to construct H5N1-recombinant FWPV (rFWPV) coexpressing chicken Interleukin 12 (IL-12) or Interleukin 15 (IL-15), and discusses the immunogenicity of the recombinants following inoculation into specific-pathogen-free (SPF) chickens. Previously cloned and sequenced cDNAs encoding full-length H5 and N1 of influenza strain A/Chicken/Malaysia/5858/2004 genes were amplified by PCR and inserted into plasmid pEFL29, under the control of a copy of the vaccinia virus p7.5 early/late promoter. The expression cassettes were recombined into the genome of the FP9 strain of FWPV at the fpv002 locus. Recombinant viruses were produced by transfection of the plasmid into chicken embryo fibroblasts (CEFs) after infection with FP9, and isolated by six fold plaque purification on CEFs using X-Gal selection. Chicken IL-12 or IL-15 genes, under control of a synthetic/hybrid poxvirus promoter, were inserted into a ‘transient dominant selection’ recombination plasmid, pPC1.X. The cytokine expression cassettes were then recombined, at the fpPC1 (fpv030) locus, into rFWPV already carrying AIV genes. Following three rounds of passage in CEFs in the presence of mycophenolic acid (MPA), recombinant viruses carrying the gpt gene were isolated. These unstable recombinants were plaque-purified in the absence of MPA until they lost the gpt gene spontaneously, verified by their failure to replicate in the presence of MPA. Recombinant proteins were successfully detected using western blotting and indirect immunofluorescence assay (IFAT). Parental and rFWPV (105 PFU) were inoculated subcutaneously into one-day-old SPF chickens. Sera from chickens immunized with rFWPV/H5 and rFWPV/H5/IL-15 demonstrated viral neutralizing activities, based on the haemagglutation inhibition (HI) test, in which reached a peak at Week 3. A competitive enzyme-linked immunosorbent (ELISA) assay detected N1-specific antibodies induced by rFWPV/N1 and rFWPV/N1/IL-12 at Weeks 4 and 5. Non-specific cellular immune responses were assessed by flow cytometric analysis to enumerate CD4+ and CD8+ T-lymphocytes in peripheral blood. Results of Experiment 2 showed chickens vaccinated with rFWPV/H5, rFWPV/H5/IL-15, rFWPV/N1 and rFWPV/N1/IL-12 demonstrated a higher increase in CD8+ than CD4+ T cell population, relative to control and chickens vaccinated with parental FWPV. Weekly weighing showed that chickens vaccinated with rFWPV/H5/IL-15 had the highest body weight compared to other groups, while the rFWPV/N1/IL-12 group showed the significantly lowest body weight. In summary, this study showed diverse immunogenicity of H5N1-rFWPV coexpressing IL-12 or IL-15. It also demonstrated a weight sparing effect of co-expressing IL-15 in rFWPV vaccines. The results provide the basis for future homologous challenge studies, using live H5N1 virus to evaluate the protective efficacy of the rFWPV vaccines

Mobile health applications digital evidence taxonomy with knowledge sharing approach for digital forensics readiness

M-health is the current application that capable to monitor and detect human biological change and used the Internet as a platform to transfer and receive the data from the cloud providers. However, the advancement of Internet of Things (IoT) technology poses a great challenge for digital forensic experts in order to preserve, acquire and analyse digital evidence. Digital evidence taxonomy is one technique in digital forensics that facilitates digital forensics readiness and integration with knowledge sharing approach is necessary to allow digital forensics experts to share their knowledge. Therefore, this research was carried out that consists three phases, namely (1) initial phase, (2) intermediate phase and (3) final phase. In the initial phase, a systematic literature review was conducted to identify any potential gaps from the existing studies. Subsequently, digital evidence taxonomy in the IoT forensics layers was adopted, which consisted of three artefact categories to represent the IoT forensics layers. In the intermediate phase, 34 top rating m-health apps were used as a case study to validate the digital evidence taxonomy. From the analysis of the result, various types of information for forensic investigation were acquired, such as type of outdoor activity, activity timestamp, client IP address and date accessed. In the final phase, the M-Health Digital Evidence Taxonomy System (MDETS) was developed as a proof of concept to demonstrate the integration of digital evidence taxonomy with the knowledge-sharing approach to facilitate digital forensic readiness. Interviews were used as the instrument tool to evaluate knowledge sharing in terms of people, process and technology elements in enabling digital forensic readiness. The results from the interviews support that knowledge sharing facilitates digital forensic readiness in terms of people, process and technology elements. As a conclusion, the integration of digital evidence taxonomy with the knowledge-sharing approach gives the opportunity for the digital forensic community to enhance the existing approach or procedure to increase the findings of a digital forensic investigation and make digital forensic readiness more proactive within the organisation

Development and evaluation of ergonomics and psycosocial assessment method for assessing work-related upper limb disorders

The observational methods have been considered as the most widely used for assessing work-related upper limb disorders (WULDs) because it is inexpensive, practical to use, and flexible to use in a wide range of workplaces. However, the existing methods have a specific purposes and do not universally ideal for all risk factors. Most of the existing methods only focus on assessing physical risk factors of WULDs especially in body postures and do not considered other factors of WULDs such as psychosocial, work organizational, and individual risk factors. Nowadays, psychosocial work characteristics are recognized as important risk factors. Besides that, during the tool development process, it was found that some of the existing observational methods have limited work perform the reliability, usability, and validity analysis. In order to improve these limitations, this study aims to develop a new observational method known as Upper Limb Risk Assessment (UPLIRA) tool and to determine the reliability and validity of the UPLIRA tool. The initial development of UPLIRA tool was conducted in three stages including the development of the UPLIRA system components, development of the UPLIRA checklist (prototype), and psychometric properties evaluation (reliability and validity). In reliability trials, the results of inter and intra rater reliability shows good (K = 0.802 - 0.806) to very good (K = 0.811 - 0.833) agreement among the observers. Meanwhile, the usability trials shows that the UPLIRA tool was applicable to wide range of jobs/tasks, valuable at works, and action level is easy to understand (mean 4.17 ± 0.753). Whereas, in validity trials the highest risk scores were obtained by cashier workers contrast to office workers according to the UPLIRA assessment. Based on the results so far, it can be concluded that the UPLIRA tool was found to be practically reliable and applicable to a wide range of workplaces

Assessment of ecosystem integrity of lowland dipterocarp forest ecosystem using remote sensing

Ecosystem Integrity Index (EII) is a concept to determine the quality or the health of an ecosystem. The EII development can assist forest managers and decision makers in the conservation effort and forest management in Malaysia through the development of a simple and easy-to-adopt index. The aim of this study is to assess and evaluate the EII through the development of forest structure empirical models from remotely sensed data for lowland dipterocarp forest in Malaysia. The objectives of this study are: (i) to assess the structure and composition of lowland dipterocarp forest in Malaysia, (ii) to develop empirical model for estimating stand structure from remotely sensed data, and (iii) to derive the ecosystem integrity index for lowland dipterocarp forest. Tree Basal Area (BA), aboveground biomass (AGB) and volume plot from plot data were used as dependent variables, while remote sensing data from Landsat, Pleiades and LiDAR were used as independent variables for model development. Tree plot census was carried out from 17 to 19 May 2016, while remote sensing data acquisition dates for Landsat, Pleiades and LiDAR were 13 March 2016, 24 January 2015 and April 2015 respectively. Forest Structure Modeling was carried out by means of a correlation analysis with the calibration of dependent and independent data to select the most significant and accurate remote sensing variables to derive empiric equation (model), fitting stage to select the best model with the highest coefficient of determination (R2) and the lowest root mean square error ( RMSE) validation of the final selected. The Ecosystem Integrity Index was developed by the average percentage of the predicted BA, AGB and model volume. The EII was categorised at five integrity levels as high (81–100%), medium high (61–80%), moderate (41–60%), medium low (21–40%) and low (0–20%). A total of 1035 trees with diameter at breast height (DBH) of 5.0 cm and above were recorded in 69.115 ha sampling areas. The total trees recorded represented 150 species from 87 genera and 34 families. Shorea macroptera (Dipterocarpaceae), S. leprosula (Dipterocarpaceae) and S. parviflora (Dipterocarpaceae) are three dominant species, with Species Important Value Index (SIVi) of 6.49%, 6.23% and 5.51%, respectively. Dipterocarpaceae is the most dominant with Family Important Value Index (FIVi) of 33.54%. The developed final model is robust and consistent with high R2 with range of 0.84 to 0.87. The final models constructed for AGB, BA and volume value of R2 are 0.85, 0.84 and 0.87 respectively. The RMSE of AGB, BA and volume model are 53.1 Mg/ha, 3.54 m2/ha and 46.4 m3/ha, respectively. The overall stand AGB, BA and volume for Sungai Menyala Forest Reserve is 282.29 Mg/ha, 17.68 m2/ha and 239.51 m3/ha. An Ecosystem Integrity Index (EII) assessment has been successfully demonstrated by this study with production of practical, multi-scaled, flexible, adjustable and policy-relevant index. The overall EII of Sungai Menyala Forest Reserve is in Category 3, which shows that the area is within the medium value

Building trustworthy e-Commerce wesite

The process of building consumer trust in E-Commerce is based on the presence of trust features or trust attributes in the websites. Consumer may examine e-Commerce websites for the existence of trust attributes. However, to date, which trust attributes contribute to the website’s trustworthiness and which trust attributes give more value to consumer has not been adequately explored. Therefore, the purpose of the paper is to look for the relevant trust attributes for e-Commerce websites and to identify the importance ranking of trust attributes that contribute significantly to the trustworthiness of e-Commerce website. Various journal papers and articles related to e-Commerce field have been referred in order to identify the trust attributes. An online survey that received 1230 respondents was carried out to investigate the importance ranking of ten trust attributes. The paper contributes to the discussion on how to build trust in e-Commerc

Removal of endocrine disrupting chemicals using low pressure reverse osmosis membrane

Endocrine disrupting chemicals (EDCs) are one of the major focuses of contaminants in current environmental issues, as they can cause adverse health effects on animals and human, particularly to endocrine function. The objective of this study was to remove a specific group of EDCs (i.e molecular weight range 228 to 288 g/mol) using low pressure reverse osmosis membrane (LPROM). A multi-layer thin-film composite of aromatic polyamide (ES20) membrane and a C10-T cross flow module of LPROM manufactured by Nitto Denko Company was used in this study. The effects of operating parameters, i.e. pH, operating pressure, concentration and temperature were observed using a design of experiment based on MINITABTM software. The analysis of results was conducted by factorial analysis (FA) and response surface analysis (RSA). It was found that LPROM has been effectively applied to remove pentachlorophenol (PCP) (more than 83%), 17ß-estradiol (more than 87%) and bisphenol-A (BPA) (more than 87%). For permeate flux, both PCP and 17ß-estradiol tests produce excellent flux rate; i.e. 23.8 L/m2.h and 22.9 L/m2.h, respectively. For BPA, the permeate flux produced was slightly lower (19.1 L/m2.h) due to its physical-chemical properties effect at various levels of the recovery rate. In this study, the percentage of rejection was increased with the increased of pH and concentration of compounds. The flux was observed to increase with the increase of operating pressure. This study also investigated the interaction effects between operating parameters involved. In addition, statistical models were developed to represent the performance of LPROM under two response parameters, i.e. percentage of EDCs rejection and permeate flux. Statistical models were then validated using One-Factor-At-a-Time (OFAT) design of experiments and comparisons were made to better understand the trend of EDCs rejection and permeate flux

Beach response due to the pressure equalization modules (PEM) system

Coastal erosion is a significant problem with dramatic effects on the coastline. There is an urgent need to introduce new and cost-effective measures that can mitigate the impacts on the shoreline. This study has been initiated to investigate the response of the beach at Teluk Cempedak due to the beach nourishment and Pressure Equalization Modules (PEM) system. The objectives of this study are the determination of closure depth and effectiveness of the system in treating the erosion process. The depth of closure was examined using both data from a series of beach profile surveys and from empirical formulae. The widely accepted Fixed Depth Change (FDC) method was explored and the hc before and after the installation of PEM system was investigated. The research found that multiple closure points can occur along the profile lines. The closure depth after the installation of PEM system was found to be deeper and the closure point is further seaward at the southern part of the beach. The Hellemeier’s equation over predict hc by 76 %, however it reveals that the equation is still robust in determining an upper limit of hc. The simplified equation was developed at Teluk Cempedak beach in predicting closure depth and can be equated to 0.98 times H0.137. From the survey data, it is found that after three years, the total sand volume and beach elevation are significantly higher in PEM areas. Generally, the result presented indicates the decreasing value of rate of erosion. Thus it revealed that PEM system is able to stimulate accretion of sand and yet slow down the erosion process. However, based on the sand volume distribution pattern, after three years, it is obviously seen that the accretion of sand occurring at the northern part while erosion process is taking place in the southern part of the beach. Based on the distribution pattern of bed elevation over the chainage, overall, the upper part of the beach is convex unlike earlier i.e before the installation of PEM system, where the beach was low and concave. This phenomena indicates that the system contribute to a significant accretion of sand and thus created a higher beach level at about 10 m to 55 m towards the sea. However, this trend only can be seen at a certain chainage. The PEM efficiency in terms of increment in bed elevation can only be observed at CH 400 till CH 800 while at CH 900 towards the south, the efficiency is decreasing. This shows that the accretion of sand is only occurring at the northern part and the beach is eroding at the southern part. Therefore, based on the available four years record of data, there is a certain part of the beach benefiting from the PEM system. However, some parts are still experiencing the erosion process

Innovative technologies for industrial wastes

This article is intended to provide several case studies of successful waste management of a few selected industries in their attempts to become “environmental-conscious” firms. In particular, this article examines the innovative waste-reduction and waste reuse processes undertaken by certain firms in the following industries - asphalt cement and concrete, ferrous metals, Portland cement and concrete, and some other that on the face of it somewhat isolated innovative technologies. For each case, the driver, the waste management technology or processes involved, as well as the associated economic benefits of the adjustments was highlighted. It is hoped that the findings of this article will provide the motivation or continue to motivate engineers and scientists to further explore processes that will help towards better management of industrial wastes