Investigation of Interactions of the Rubella Virus P150 Replicase Protein with Host Cell Proteins in Infected Cells

Due to their simplicity, viruses require the assistance of host factors for various aspects of their replication cycle. This study investigated the interaction of one of the two non-structural replicase proteins of rubella virus (RUBV), P150, with cell proteins. RUBV forms replication complexes for replicating its RNA in association with membranes of endosomes and lysosomes; the thusly modified endosomes/lysosomes are termed cytopathic vacuoles or CPVs. In the first study, a RUBV expressing a FLAG epitope-tagged P150 was used to co-immunoprecipitate putative interacting cell proteins from an infected cell lysate fraction enriched for CPVs using differential centrifugation. However, the only interacting protein identified was the companion RUBV replicase protein P90. Thus, cell proteins do not bind with either sufficient affinity or in stoichiometric amounts to be detected by this method and may not be a component of the virus holoenzyme. In the second study, a proline-rich region within P150 with three PxxPxR consensus SH3 domain-binding motifs was investigated for its ability to bind cell proteins. Substitution mutations (to alanine) of the two prolines were made in each of these motifs with the finding that mutations in the first two motifs led to lower viral titers and a small plaque phenotype with reversion to the wt sequence within one passage. Mutations in the third motif had a wt phenotype and did not revert. However, these mutations did not affect viral RNA synthesis, suggesting that the importance of these motifs is in a later stage of viral life cycle, e.g. virion assembly and release. To extend these findings, the proline hinge region with either the wt or mutant sequence was expressed as a GST-fusion in human cells. Pulldown experiments revealed specific binding with human p32 protein (gC1qR), which was previously shown to interact with the RUBV capsid protein. Binding of p32 with P150 was confirmed. The function of p32 in the RUBV replication cycle is unclear, but could involve virion assembly and release or induction of apoptosis

History of moneylending during British era: A case study of Chettiars as the major money lenders

This paper aims to study history of moneylending during the British era by focusing on Chettiars as the professional moneylenders. Money lending is synonym with the name of Chettiars or ‘Chetty’ in Malaysia.Notably, Chettiars were the major moneylenders during the era of British from late nineteenth century until Independence of Federation of Malaya in 1957. Chettiars, a Tamil speaking business caste from South India migrated to Straits Settlements during nineteenth century to make fortune in trade.Gradually, their long tradition and experiences as indigenous banking caste and trade on open market created opportunities for them to involve in the money transactions in the Straits Settlements.Lack of credit and bank facilities made the local and foreign traders at the the Straits Settlements to rely on the Chettiar agents. Role of Chettiars became more prominent after the late nineteenth century on wards due to development of commercial economy such as rubber and tin under British laissez faire policy in the Malay states.Yet, neither banking nor British credit facilities could cater/accommodate the fast growing commercial economy in Malay states.Thus, the local and Asian capitalists relied on loans from Chettiar for their investments to open up thick virgin jungles for rubber cultivation and tin mining.Large capital and liberal lending methods resulted Chettiars to be known as leading money lenders in the Malay States.Through moneylending activities, Chettiars also played other roles in the socioeconomy development of Malaya.However, borrowing became rife especially among the small capitalists (farmers, tin miners and petty traders) and low income civil servants during the recession period and reached alarming stage whereby British realized the need to curb money lending activities of Chettiars.Thus, British implemented laws and administrative regulations which were later on proof to be unsuccessful until World War II. After the Japanese occupation, British became more tactful towards Chettiars until more stringent law was enforced to control money lending activities in Malaya. Apparently, The Moneylenders Bill, 1951 was the first of its kind/legislation and created greater dis-satisfactions among the Chettiar moneylenders which they foresee as the end for their money lending activities in Malaya.Within this context, this paper reveals on the history of money lending activities during the British era from glorious period of the Chettiars’ role as the leading money lenders until end of their moneylending activitie

The Past, Present and Future of Diagnostic Imaging in Ovarian Cancer

Ovarian cancers (OC) include a group of diseases with variable prognoses. While most conventional imaging techniques rely on the detection of tumour burden and distant spread to identify treatment plans, more emphasis is now being placed on screening for early detection and also for more accurate staging using molecular imaging techniques. It is generally accepted that there are some incremental benefits of using serum CA125 levels coupled with cross-sectional diagnostic imaging to aid in the diagnosis, staging and treatment planning of OC. This chapter provides a review of tests and diagnostic imaging modalities that aid in the detection and staging of OC with a particular focus on F18-Fluorodeoxyglucose positron emission tomography/computed tomography (F18-FDG PET/CT) imaging. This chapter also proposes a diagnostic algorithm for the management of ovarian cancer. F18-FDG PET/CT imaging can act as a catalyst for the development of personalised medicine by stimulating advancements in targeted therapy. In conclusion, diagnostic imaging with particular focus in molecular imaging has the potential for altering management plans, which can ultimately help improve the prognosis of ovarian cancer

Lack of Processing of the Expressed ORF1 Gene Product of Hepatitis E Virus

Background: Proteolytic processing is a common mechanism among plus strand RNA viruses and the replicases of all plus strand RNA viruses of animals thus far characterized undergo such processing. The replicase proteins of hepatitis E virus (HEV) are encoded by ORF1. A previous report published by our group [1] provided data that processing potentially occurred when ORF1 (Burma strain; genotype 1) was expressed using a vaccinia virus-based expression system. Findings: To further test for processing and to rule out artifacts associated with the expression system, ORF1 was re-expressed using a plasmid-based expression vector with the result that the previous processing profile could not be confirmed. When ORF1 from an HEV infectious cDNA clone (US swine strain; genotype 3) was expressed using the plasmid-based system, the only species detected was the 185 kDa precursor of ORF1. A putative papain-like cysteine protease [2] had been predicted within ORF1 using the original HEV genomic sequence. However, analysis of subsequent ORF1 sequences from a large number of HEV isolates reveals that this protease motif is not conserved. Conclusions: The expressed HEV ORF1 gene product does not undergo proteolytic processing, indicating that the replicase precursor of HEV is potentially unique in this regard

Conflict Management Styles Among Public Sector Managers in Malaysia.

This study examined the competence-based approach to conflict management from the Malaysian perspective especially on the effectiveness and appropriateness of the conflict management styles as perceived by dyads of superiors-subordinates. In the process, this study set out to determine the conflict management styles used by Administrative and Diplomatic Services (ADS) officers in handling interpersonal conflicts with their subordinates. In addition, it looked at the moderating effects of individual attribute, professional profile and management styles on the effectiveness and appropriateness of the conflict management styles used as reported by the superiors and subordinates. Self-administered questionnaires were used to gather data. The sample consisted of 383 dyads of superiors-subordinates selected randomly from the 28 Ministries/Department. The quantitative data were analysed using various statistical and exploratory data analysis methods, descriptive tests, ANOVAs, Chi-square test of independence, and MANOVA.The findings indicated that ADS officers used integrating, compromising and dominating styles to handle interpersonal conflicts with their subordinates. The least used style was the obliging style. The 9,9 management style was the most frequently used style followed by 1,9 management style. The least used style was the 1 ,I management style. Both the superiors and subordinates rated the integrating style as the most effective, most relationally appropriate and most situationally appropriate style. The obliging style was rated as the least effective and least situationally appropriate style by both the superiors and subordinates while the dominating style was reported as the least relationally appropriate. The findings revealed that the subordinates were relatively satisfied with the level of supervision. However, there were no significant findings on the moderating effects of individual attribute, professional profile and management styles on the effectiveness and appropriateness of the conflict management styles used as reported by superiors and subordinates. Further research was recommended, in particular on the competence-based approach to conflict management involving a social setting within an Eastern environment

Impact of Electronic Flight Bag on Pilot Workload

The increase of automation in the aviation industry pose challenges to human performance. To attest this point, studies about aircraft accidents reveal that pilots’ response to automated systems are always not coherent. Research findings suggests that pilots’ interaction with automated systems in highly demanding tasks situations results in the increase in workload and if they are unable to resolve it in time, it will compromise flight safety. Therefore, in the interest to further explore the impact of automation on human factor constructs, the study aimed to investigate the impact of Electronic Flight Bag (EFB) on pilot workload. The study measured the workload experienced by pilots in a visual flight rule approach in expected and unexpected situations with the use of EFB and paper chart displays. The National Aeronautics and Space Administration -Task Load Index was used to measure pilot workload. The results showed a significant difference in pilot workload between expected and unexpected approach indicating the influence of pilot workload during highly demanding tasks. However, there was no significant difference in pilot workload between the use EFB and paper at approach. There was also no significant interaction between approach and display. It is suggested that future studies to increase the sample size and explore more demanding flight situations that allows further use of EFB functionalities

Humoral immunity in colorectal cancer : evaluation of the anti-p53 and anti-hTERT auto-antibody responses

Carcinogenesis is a multi-factorial and multi-aetiological process involving suppressions, alterations and re-activation of key biomolecular markers. Some of these changes are recognised by the humoral system and are known as Tumour Associated Antigens (TAA) against which the humoral system is able to mount an auto-antibody response. Cancer cells are subject to two key mortality barriers (M1 & M2) as described in the "2-hit" hypothesis which are overcome by p53 dysfunction (M1) and hTERT re-expression (M2). These two molecular events generate TAA which are recognised by the humoral immune system with a corresponding auto-antibody response. The aim of this thesis was to investigate the significance of the humoral anti-p53 auto-antibody and anti-hTERT auto-antibody responses in colorectal cancer (CRC). This was performed by evaluating all published literature (1979-2009) on anti-p53 auto-antibody response and its association with p53 mutation to provide the largest cumulative sample size to date spanning 30 years. A critical review was performed of all anti-p53 auto-antibody studies in CRC, followed by an investigation into the long-term prognostic significance (minimum 5 years follow-up) of anti-p53 auto-antibody in CRC. The second aim of this thesis was to optimise a method of detecting anti-hTERT in CRC patients and correlate this with anti-p53 auto-antibody in order to investigate the significance of a joint humoral response against the two key TAA responsible for CRC immortality.The overall prevalence of anti-p53 auto-antibody was 18.4% (3292/17,859) in all cancers and 2.2% (88/3,946) in normal/benign disease controls. The anti-p53 autoantibody presence in all published cancers reports was plotted against the reported p53 mutational rates in individual cancers and showed partial correlation (R2=0.5, correlation=0.7) between anti-p53 auto-antibody presence and p53 mutation. Anti-p53 was present in 21.5% (786/3,653) in all CRC only studies, and 19.9% (479/2,409) in CRC studies using ELISA. Anti-p53 was not associated with clinico-pathological factors or prognosis in majority of the studies. Only 4 studies associate anti-53 autoantibody with adverse clinico-pathological parameters, mostly in selective groups. The weaknesses of these studies are discussed. This association leads to anti-p53 association with adverse prognosis but only in selective analysis. The prognostic significance is observed in univariate analysis but lost in multivariate analysis when stronger traditional prognostic factors are incorporated.This thesis initially compared serum with plasma anti-p53 auto-antibody titres and excluded plasma titres from further analysis due to the potential contamination by non-specific binding leading to falsely elevated levels (17-73% variation) of anti-p53 auto-antibody. Serum anti-p53 auto-antibody was present in 21.7% (20/92) CRC patients and 0% (0/20) controls. There was no association with age (p=0.750), sex (p-0.468), Dukes' / TNM stage (p=1.000), T- (p=0.900), N- (p=0.912), M-stage (p=0.632), location (p=0.175), differentiation (p=0.117) or mucinous component (p=0.699). The median follow-up was 97 months with median DPS and OS of 73 months and 62 months respectively. Dukes' / TNM stage, T-, N-, M-stage were prognostic indicators in univariate DFS and OS analysis. Only Dukes'/TNM stage remained an independent prognostic indicators in multivariate analysis (p=0.001). Anti-p53 auto-antibody did not display prognostic significance in univariate or multivariate analysis of OS or DFS.Anti-hTERT auto-antibody has only been reported once in the literature, using molecular recombination to develop hTERT antigen. This thesis optimisation processes aimed develop a method of detecting anti-hTERT using less restrictive technology, and further development of a WB or ELISA to allow mass detection of serum anti-hTERT. The first step aimed to isolate hTERT using a streptavidin immune-affinity column with biotinylated anti-hTERT to capture hTERT from cancer cell lysates. This was unsuccessful and further attempts were made at identifying hTERT using Western blot (WB). Five different anti-hTERT antibodies and a multitude of WB conditions were trialled in duplicate (>100 WB), each with multiple ECL exposures. hTERT was not identified. The reason for this was narrowed down in the final experiments to the lack of specificity of the primary antibodies available. The raising of a sufficiently specific anti-hTERT antibody is required to isolate hTERT antigen.Early CRC detection is vital in improving outcomes. The humoral response to TAA incarcinogenesis could enable earlier identification of CRC and impact prognosis

Effects of Cognitive Stress on the Temporal Anticipation of a Timing Motor Task

The study was designed to investigate the effects of cognitive stress on the temporal anticipation of a timing motor task. The experimental design applied was a repeated measures design with two independent variables (cognitive stress and levels of difficulty - easy, intermediate and difficult). The participants of this study consist of 18 male and 19 female undergraduates of the Physical Education programme of University Putra Malaysia. The participants were required to perform a timing motor across three levels of difficulty and under two conditions i.e. without cognitive stress and under cognitive stress. Cognitive stress was induced via the continuous subtraction of two from a two-digit number. Participants performed the task individually and the sequence of the experimental task was counterbalanced. A two-way within subject's ANOVA was performed to ascertain the effects of cognitive stress on the temporal anticipation of the timing motor task. The data analysis revealed a significant difference in means for the stress main effect (A = 64, E (1.35) = 19.89, P = .00); and the task main effect (A = .84, E (2,34) = 3.35, p < .05). Post hoc comparisons produced a significant difference in the means of the performance of the timing motor task at all three levels of difficulty. This showed that cognitive stress had an effect on the temporal anticipation of the timing motor task. A two way mixed design ANOVA was conducted was performed to ascertain the effects of cognitive stress on the temporal anticipation of the timing motor task with regard to gender. The task main effect was significant (A = .59, E (5, 170) = 6.70, P = .01). Follow up comparisons revealed significant difference in the performance of the experimental task at the easy level without cognitive stress, intermediate level without cognitive stress and the easy level under cognitive stress. This showed that there was a significant difference in the performance of the timing motor at the three levels mentioned

Gorham's disappearing bone disease: a rare cause of extensive bone destruction

Osteolytic disease of the bones have a myriad range of aetiology. One rare cause is Gorham’s disease or disappearing bone disease. This disease is a diagnosis by exclusion using correlation made with clinical presentation, radiological findings and histopathological confirmation. Although many different therapies have been advocated, none have been successful in fully controlling this disease. We present a case that was detected in a Chinese lady using diagnostic imaging, confirmed with computed tomography guided biopsy and successfully treated with joint reconstruction using endoprosthesis

Upper Limb Movement Recognition utilising EEG and EMG Signals for Rehabilitative Robotics

Upper limb movement classification, which maps input signals to the target activities, is a key building block in the control of rehabilitative robotics. Classifiers are trained for the rehabilitative system to comprehend the desires of the patient whose upper limbs do not function properly. Electromyography (EMG) signals and Electroencephalography (EEG) signals are used widely for upper limb movement classification. By analysing the classification results of the real-time EEG and EMG signals, the system can understand the intention of the user and predict the events that one would like to carry out. Accordingly, it will provide external help to the user. However, the noise in the real-time EEG and EMG data collection process contaminates the effectiveness of the data, which undermines classification performance. Moreover, not all patients process strong EMG signals due to muscle damage and neuromuscular disorder. To address these issues, this paper explores different feature extraction techniques and machine learning and deep learning models for EEG and EMG signals classification and proposes a novel decision-level multisensor fusion technique to integrate EEG signals with EMG signals. This system retrieves effective information from both sources to understand and predict the desire of the user, and thus aid. By testing out the proposed technique on a publicly available WAY-EEG-GAL dataset, which contains EEG and EMG signals that were recorded simultaneously, we manage to conclude the feasibility and effectiveness of the novel system.Comment: 20 pages, 11 figures, 2 tables; Thesis for Undergraduate Research Project in Computing, NUS; Accepted by Future of Information and Communication Conference 2023, San Francisc