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Fulltext Validation of a simple extraction procedure for bisphenol A identification from human plasma

Wiraagni IA, Mohd MA, Bin Abd Rashid R, Haron DEBM

PLoS One, 2019;14(10):e0221774.
PMID: 31581189 DOI: 10.1371/journal.pone.0221774

The general population is exposed to bisphenol A (BPA) orally, parenterally, transdermally, and environmentally as a result of the use of BPA in food packaging, plastics, and personal care products. The majority of the population nowadays (91-99%) has detectable levels of BPA inside their body. In this study, we successfully performed an inexpensive, rapid, and simple protein precipitation procedure for extraction of BPA from human plasma, followed by analysis by LC-MS/MS. This method was specifically developed for handling large numbers of samples with minimum cost and volume of sample. The developed method was accurate, precise, and reproducible for quantification of BPA from human plasma samples in the concentration range of 10-2000 ng/mL. The method was performed on samples from 150 healthy volunteers who were enrolled in the study. The mean of observed BPA level was 2.22 ± 9.91 ng/mL. Higher BPA levels were observed for females compare to that of males (p-value = 0.002), the BPA levels were higher in participants 33 years of age and older compared to those less than 33 years of age (p-value = 0.000), then the BPA levels higher in subjects with tap water as source of drinking (p-value = 0.005). This method may be valuable for general risk assessment of BPA for a large and varied population because of its efficiency and economical aspects.
Fulltext Validation of a Malay Version of the Smartphone Addiction Scale among Medical Students in Malaysia

Ching SM, Yee A, Ramachandran V, Sazlly Lim SM, Wan Sulaiman WA, Foo YL, et al.

PLoS One, 2015;10(10):e0139337.
PMID: 26431511 DOI: 10.1371/journal.pone.0139337

This study was initiated to determine the psychometric properties of the Smart Phone Addiction Scale (SAS) by translating and validating this scale into the Malay language (SAS-M), which is the main language spoken in Malaysia. This study can distinguish smart phone and internet addiction among multi-ethnic Malaysian medical students. In addition, the reliability and validity of the SAS was also demonstrated.
Fulltext Validation of CHERG'S Verbal Autopsy-Social Autopsy (VASA) tool for ascertaining determinants and causes of under-five child mortality in Pakistan

Siddiqui MB, Ng CW, Low WY, Abid K

PLoS One, 2023;18(12):e0278149.
PMID: 38109305 DOI: 10.1371/journal.pone.0278149

The majority (40%) of the world's under-five mortality burden is concentrated in nations like Nigeria (16.5%), India (16%), Pakistan (8%), and the Democratic Republic of the Congo (6%), where an undetermined number of under-five deaths go unrecorded. In low-resource settings throughout the world, the Verbal Autopsy-Social Autopsy (VASA) technique may assist assess under-five mortality estimates, assigning medical and social causes of death, and identifying relevant determinants. Uncertainty regarding missing data in high-burden nations like Pakistan necessitates a valid and reliable VASA instrument. This is the first study to validate Child Health Epidemiology Reference Group-CHERG's VASA tool globally. In Pakistan, data from such a valid and reliable tool is vital for policy. This paper reports on the VASA tool in Karachi, Pakistan. Validity and reliability of the CHERG VASA tool were tested using face, content, discriminant validation, and reliability tests on one hundred randomly selected mothers who had recently experienced an under-five child death event. Data were computed on SPSS (version-21) and R software. Testing revealed high Item-content Validity Index (I-CVI) (>81.43%); high Cronbach's Alpha (0.843); the accuracy of between 75-100% of the discriminants classifying births to live and stillbirths; and I-CVI (>82.07% and 88.98% respectively) with high accuracy (92% and 97% respectively) for assigning biological and social causes of child deaths, respectively. The CHERG VASA questionnaire was found relevant to the conceptual framework and valid in Pakistan. This valid tool can assign accurate medical and non-medical causes of child mortality cases occurring in Pakistan.
Fulltext Validation of Brunei's Malay EQ-5D Questionnaire in Patients with Type 2 Diabetes

Koh D, Abdullah AM, Wang P, Lin N, Luo N

PLoS One, 2016;11(11):e0165555.
PMID: 27835652 DOI: 10.1371/journal.pone.0165555

BACKGROUND: The Malay spoken in Brunei a South East Asian country where Malay is the national language is distinctive and different from Malay spoken in Malaysia, Singapore and Indonesia. This study aimed to develop a Brunei Malay version of the 5-level EQ-5D questionnaire (EQ-5D-5L) and to assess its psychometric properties among patients with type 2 diabetes mellitus (T2DM).
METHODS: The Brunei Malay EQ-5D-5L was developed by culturally adapting two existing Malay versions. A total of 154 Bruneians with T2DM completed the questionnaire in two different points of time with one week apart. Known-groups validity of the utility-based EQ-5D-5L index and visual analogue scale (EQ-VAS) was evaluated by comparing subgroups of patients known to differ in health status. Test-retest reliability was assessed using the intraclass correlation coefficient (ICC) or Cohen's kappa.
RESULTS: As hypothesized, patients known to have 'better' health had higher EQ-5D-5L index scores than those having 'worse' health in all 7 known-groups comparisons. The hypothesized difference in the EQ-VAS scores was observed in only 4 of the 7 known-groups comparisons. Kappa values ranged from 0.206 to 0.446 for the EQ-5D-5L items; the ICC value for the EQ-5D-5L index and EQ-VAS was 0.626 and 0.521, respectively.
CONCLUSIONS: The utility-based EQ-5D-5L index appears to be valid and reliable for measuring the health of Brunei patients with T2DM. The validity of the EQ-VAS in Brunei requires further investigation.
Fulltext Validation and the associated factors of the Malay version of systemic lupus erythematosus-specific health-related quality of life questionnaires (SLEQoL and LupusQoL)

Ahmad Pouzi NA, Shaharir SS, Mohd Tamil A, Mustafar R, Ahmad Maulana S, Mageswaren E, et al.

PLoS One, 2023;18(5):e0285461.
PMID: 37186611 DOI: 10.1371/journal.pone.0285461

OBJECTIVES: To assess the reliability and validity of two disease-specific questionnaires that assess the quality of life (QoL) among patients with Systemic Lupus Erythematosus (SLE); SLEQoL and LupusQoL in Malay language. This study also identified the factors affecting each domain of the questionnaires.
METHODS: This cross-sectional study was conducted from June 2021 until April 2022, and SLE patients were recruited to complete the SLEQoL, LupusQoL and Short Form Health Survey (SF-36) in Malay language. Disease activity were recorded using the modified SLE Disease Activity Index (M- SLEDAI) and British Isles Lupus Assessment Group 2004 (BILAG-2004) index. Presence of organ damage was determined using the SLICC Damage index. Cronbach's alpha was calculated to determine internal consistency while exploratory factor analysis was done to determine the construct validity. Concurrent validity was evaluated using correlation with SF-36. Multiple linear regression analysis was deployed to determine the factors affecting each domain of SLEQoL and LupusQoL.
RESULTS: A total of 125 subjects were recruited. The Cronbach's α value for the Malay-SLEQoL (M-SLEQoL) and Malay-LupusQOL (M-LupusQoL) was 0.890 and 0.944 respectively. Exploratory factor analysis found formation of similar number of components with the original version of questionnaires and all items have good factor loading of >0.4. Both instruments also had good concurrent validity with SF-36. M-SLEQoL had good correlations with BILAG-2004 and M-SLEDAI scores. Musculoskeletal (MSK) involvement was independently associated with lower M-SLEQoL in physical function, activity and symptom domains. Meanwhile, MSK and NPSLE were associated with fatigue in M-LupusQoL.
CONCLUSION: Both M-SLEQoL and M-LupusQoL are reliable and valid as disease -specific QoL instruments for Malaysian patients. The M-Lupus QoL has better discriminative validity compared to the M-SLEQoL. SLE patients with MSK involvement are at risk of poor QoL in multiple domains including physical function, activity, symptoms and fatigue.
Fulltext Validation and reliability of the Bahasa Malaysia language version of the Acceptance of Illness Scale among Malaysian patients with cancer

Leong WC, Azmi NA, Wee LH, Rajah HDA, Chan CMH

PLoS One, 2021;16(9):e0256216.
PMID: 34587199 DOI: 10.1371/journal.pone.0256216

Cancer is a life-threatening disease, and the challenges in accepting the diagnosis can bring a devastating emotional impact on the patient's mental and psychological wellbeing. Issues related to illness acceptance among cancer patients are not well studied in Malaysia. To date, the Acceptance of Illness Scale has not been translated to the Malay language (Bahasa Malaysia) nor validated for use in the oncology setting. The objective of the study is to translate, validate and determine the reliability of the Bahasa Malaysia version of the Acceptance of Illness Scale among Malaysian patients with cancer. A total of 129 patients newly diagnosed with cancer were consecutively sampled and the scale was administered via face-to-face interviews. A pilot test (n = 30) was conducted and test-retest reliability was determined. The Bartlett Test of Sphericity was statistically significantly (p<0.001), while the Kaiser-Mayer-Olkin (KMO) measure of sampling adequacy was adequate at 0.84. Scale item mean scores ranged between 3.02 and 4.33, while the item-total correlation ranged between 0.50 to 0.66 (p<0.05). The internal reliability coefficient was 0.84. The test-retest reliability indicated a high correlation, r = 0.94 with p = 0.001. The Bahasa Malaysia version of the Acceptance of Illness Scale is a valid and reliable instrument that is appropriate for use in Malaysian patients with cancer. Use of this scale to assess illness acceptance among the Malay-speaking patients with cancer can act as a guide for delivery of psycho-oncological services to help patients have a better mental wellbeing and life adjustment in living with cancer.
Fulltext Vacuum/compression valving (VCV) using parrafin-wax on a centrifugal microfluidic CD platform

Al-Faqheri W, Ibrahim F, Thio TH, Moebius J, Joseph K, Arof H, et al.

PLoS One, 2013;8(3):e58523.
PMID: 23505528 DOI: 10.1371/journal.pone.0058523

This paper introduces novel vacuum/compression valves (VCVs) utilizing paraffin wax. A VCV is implemented by sealing the venting channel/hole with wax plugs (for normally-closed valve), or to be sealed by wax (for normally-open valve), and is activated by localized heating on the CD surface. We demonstrate that the VCV provides the advantages of avoiding unnecessary heating of the sample/reagents in the diagnostic process, allowing for vacuum sealing of the CD, and clear separation of the paraffin wax from the sample/reagents in the microfluidic process. As a proof of concept, the microfluidic processes of liquid flow switching and liquid metering is demonstrated with the VCV. Results show that the VCV lowers the required spinning frequency to perform the microfluidic processes with high accuracy and ease of control.
Fulltext Utilization of magnesium-rich synthetic gypsum as magnesium fertilizer for oil palm grown on acidic soil

Ayanda AF, Jusop S, Ishak CF, Othman R

PLoS One, 2020;15(6):e0234045.
PMID: 32544208 DOI: 10.1371/journal.pone.0234045

A study was conducted to determine the impact of applying different sources of Mg, namely kieserite, ground magnesium limestone (GML) and Mg-rich synthetic gypsum (MRSG) on an acid tropical soil, oil palm growth and production. Besides high amount of Mg and Ca, MRSG contains S. Exchangeable Ca in the untreated soil of the plantation was 0.64 cmolc kg-1, but its critical level to sustain oil palm growth was 0.9 cmolc kg-1. MRSG was applied in the plantation as Mg-fertilizer; however, since Ca is also a limiting nutrient, oil palm growth was correlated (r = 0.69) with Ca supplied by the MRSG. Mg needed to sustain oil palm production is normally supplied by kieserite. Its requirement can be met at a lower cost compared to that of the kieserite by using MRSG. Due to MRSG treatment, exchangeable Ca in the soil increased steadily to satisfy the requirement of oil palm for fruit bunches production. From the glasshouse and field study, it was observed that MRSG applied at 1.5 times the recommended rate gave results comparable to that of the kieserite. MRSG treatment resulted in the increase of soil pH to >5 that precipitated Al3+ as inert Al-hydroxides, which eventually enhanced oil palm seedlings growth. Thus, MRSG can also replace GML to increase soil pH and satisfy the Ca and Mg requirement of oil palm. It can be concluded that MRSG has the potential to be used as a source of Mg as well as Ca for oil palm grown on acidic soil.
Fulltext Utilization of health care services for childhood morbidity and associated factors in India: a national cross-sectional household survey

Sreeramareddy CT, Sathyanarayana TN, Kumar HN

PLoS One, 2012;7(12):e51904.
PMID: 23284810 DOI: 10.1371/journal.pone.0051904

BACKGROUND: Information about utilization of health services and associated factors are useful for improving service delivery to achieve universal health coverage.
METHODS: Data on a sample of ever-married women from India Demographic and Health survey 2005-06 was used. Mothers of children aged 0-59 months were asked about child's illnesses and type of health facilities where treatment was given during 15 days prior to the survey date. Type of health facilities were grouped as informal provider, public provider and private provider. Factors associated with utilization of health services for diarrhea and fever/cough was assessed according to Andersen's health behavior model. Multinomial logistic regression analyses were done considering sampling weights for complex sampling design.
RESULTS: A total of 48,679 of ever-married women reported that 9.1% 14.8% and 17.67% of their children had diarrhea, fever and cough respectively. Nearly one-third of the children with diarrhea and fever/cough did not receive any treatment. Two-thirds of children who received treatment were from private health care providers (HCPs). Among predisposing factors, children aged 1-2 years and those born at health facility (public/private) were more likely to be taken to any type of HCP during illness. Among enabling factors, as compared to poorer household, wealthier households were 2.5 times more likely to choose private HCPs for any illness. Children in rural areas were likely to be taken to any type of HCP for diarrhea but rural children were less likely to utilize private HCP for fever/cough. 'Need' factors i.e. children having severe symptoms were 2-3 times more likely to be taken to any type of HCP.
CONCLUSION: Private HCPs were preferred for treatment of childhood illnesses. Involvement of private HCPs may be considered while planning child health programs. Health insurance scheme for childhood illnesses may to protect economically weaker sections from out-of-pocket health expenditure during child illness.
Fulltext Using simulation model as a tool for analyzing bus service reliability and implementing improvement strategies

Moosavi SMH, Ismail A, Yuen CW

PLoS One, 2020;15(5):e0232799.
PMID: 32379848 DOI: 10.1371/journal.pone.0232799

Bus services naturally tend to be unstable and are not always capable of adhering to schedules without control strategies. Therefore, bus users and bus service providers face travel time variation and irregularity. After a comprehensive review of the literature, a significant gap was recognized in the field of public transportation reliability. According to literature, there is no consistency in reliability definition and indicators. Companies have their own definition of bus service reliability, and they mostly neglect the passengers' perspective of reliability. Therefore, four reliability indicators were selected in this study to fill the gap in the literature and cover both passengers' and operators' perceptions of reliability: waiting time and on-board crowding level from passengers' perspective, and headway regularity index at stops (HRIS) and bus bunching/big gap percentage from operators' perspective. The primary objective of this research is to improve the reliability of high frequency of bus service and simulation tools currently being used by the public transportation companies. Therefore, a simulation model of bus service was developed to study the strategies to alleviate it. Four different types of strategies were selected and implemented according to Route U32 (Kuala Lumpur) specifications. Model out-put showed that control strategies such as headway-based dispatching could significantly improve headway regularity by almost 62% and the waiting time by 51% on average. Both holding strategies at key stops (previous and Prefol holding) have shown an almost similar impact on reliability indicators. Waiting time was reduced by 44% and 43% after the previous and Prefol Headway strategies were adopted, respectively. However, the implementation of the component of headway-based strategies at the terminal and key stops showed the best impact on reliability, in terms of passenger waiting time. Waiting time and excess waiting time were both significantly reduced by 52.86% and 81.44%, respectively. Nevertheless, the strategies did not show any significant positive effect on the level of crowding during morning peak hours.
Fulltext Using satellite-measured relative humidity for prediction of Metisa plana's population in oil palm plantations: A comparative assessment of regression and artificial neural network models

Ruslan SA, Muharam FM, Zulkafli Z, Omar D, Zambri MP

PLoS One, 2019;14(10):e0223968.
PMID: 31626637 DOI: 10.1371/journal.pone.0223968

Metisa plana (Walker) is a leaf defoliating pest that is able to cause staggering economical losses to oil palm cultivation. Considering the economic devastation that the pest could bring, an early warning system to predict its outbreak is crucial. The state of art of satellite technologies are now able to derive environmental factors such as relative humidity (RH) that may influence pest population's fluctuations in rapid, harmless, and cost-effective manners. This study examined the relationship between the presence of Metisa plana at different time lags and remote sensing (RS) derived RH by using statistical and machine learning approaches. Metisa plana census data of cumulated larvae instar 1, 2, 3, and 4 were collected biweekly in 2014 and 2015 in an oil palm plantation in Muadzam Shah, Pahang, Malaysia. Relative humidity values derived from Moderate Resolution Imaging Spectroradiometer (MODIS) satellite images were apportioned to 6 time lags; 1 week (T1), 2 weeks (T2), 3 week (T3), 4 weeks (T4), 5 week (T5) and 6 weeks (T6) and paired with the respective census data. Pearson's correlation was carried out to analyse the relationship between Metisa plana and RH at different time lags. Regression analyses and artificial neural network (ANN) were also conducted to develop the best prediction model of Metisa plana's outbreak. The results showed relatively high correlations, positively or negatively, between the presences of Metisa plana with RH ranging from 0.46 to 0.99. ANN was found to be superior to regression models with the adjusted coefficient of determination (R2) between the actual and predicted Metisa plana values ranging from 0.06 to 0.57 versus 0.00 to 0.05. The analysis on the best time lags illustrated that the multiple time lags were more influential on the Metisa plana population than the individual time lags. The best Metisa plana prediction model was derived from T1, T2 and T3 multiple time lags modelled using the ANN algorithm with R2 value of 0.57, errors below 1.14 and accuracies above 93%. Based on the result of this study, the elucidation of Metisa plana's landscape ecology was possible with the utilization of RH as the predictor variable in consideration of the time lag effects of RH on the pest's population.
Fulltext Using cascade CNN-LSTM-FCNs to identify AI-altered video based on eye state sequence

Saealal MS, Ibrahim MZ, Mulvaney DJ, Shapiai MI, Fadilah N

PLoS One, 2022;17(12):e0278989.
PMID: 36520851 DOI: 10.1371/journal.pone.0278989

Deep learning is notably successful in data analysis, computer vision, and human control. Nevertheless, this approach has inevitably allowed the development of DeepFake video sequences and images that could be altered so that the changes are not easily or explicitly detectable. Such alterations have been recently used to spread false news or disinformation. This study aims to identify Deepfaked videos and images and alert viewers to the possible falsity of the information. The current work presented a novel means of revealing fake face videos by cascading the convolution network with recurrent neural networks and fully connected network (FCN) models. The system detection approach utilizes the eye-blinking state in temporal video frames. Notwithstanding, it is deemed challenging to precisely depict (i) artificiality in fake videos and (ii) spatial information within the individual frame through this physiological signal. Spatial features were extracted using the VGG16 network and trained with the ImageNet dataset. The temporal features were then extracted in every 20 sequences through the LSTM network. On another note, the pre-processed eye-blinking state served as a probability to generate a novel BPD dataset. This newly-acquired dataset was fed to three models for training purposes with each entailing four, three, and six hidden layers, respectively. Every model constitutes a unique architecture and specific dropout value. Resultantly, the model optimally and accurately identified tampered videos within the dataset. The study model was assessed using the current BPD dataset based on one of the most complex datasets (FaceForensic++) with 90.8% accuracy. Such precision was successfully maintained in datasets that were not used in the training process. The training process was also accelerated by lowering the computation prerequisites.
Fulltext Using bootstrapped quantile regression analysis for small sample research in applied linguistics: Some methodological considerations

Nikitina L, Paidi R, Furuoka F

PLoS One, 2019;14(1):e0210668.
PMID: 30640925 DOI: 10.1371/journal.pone.0210668

Quantitative applied linguistics research often takes place in restricted settings of an intact language classroom, workplace, phonetics laboratory or longitudinal sample. In such settings the samples tend to be small, which raises several methodological problems. The main aim of the current paper is to give a detailed explanation of methodological and practical implications inherent in a robust statistical method called bootstrapped quantile regression (BQR) analysis. Importantly for applied linguistics research, the BQR method could help to deal with methodological difficulties inherent in small sample studies. The current study employed a moderately small sample (N = 27) of students learning the Japanese language in a Malaysian public university. It examined the relationships between the students' language learning motivation (specifically, integrative orientation), the students' images or stereotypes about Japan and their global attitudes toward the target language country and its people. The findings indicated that there was a statistically significant relationship between the students' attitudes toward the target language country and their integrative orientation. In addition, these attitudes were found to be the most constant determinant of the integrative orientation. Besides the applied linguistics research, the BQR method can be used in a variety of the human sciences research where a sample size is small.
Fulltext Using bioconductor package BiGGR for metabolic flux estimation based on gene expression changes in brain

Gavai AK, Supandi F, Hettling H, Murrell P, Leunissen JA, van Beek JH

PLoS One, 2015;10(3):e0119016.
PMID: 25806817 DOI: 10.1371/journal.pone.0119016

Predicting the distribution of metabolic fluxes in biochemical networks is of major interest in systems biology. Several databases provide metabolic reconstructions for different organisms. Software to analyze flux distributions exists, among others for the proprietary MATLAB environment. Given the large user community for the R computing environment, a simple implementation of flux analysis in R appears desirable and will facilitate easy interaction with computational tools to handle gene expression data. We extended the R software package BiGGR, an implementation of metabolic flux analysis in R. BiGGR makes use of public metabolic reconstruction databases, and contains the BiGG database and the reconstruction of human metabolism Recon2 as Systems Biology Markup Language (SBML) objects. Models can be assembled by querying the databases for pathways, genes or reactions of interest. Fluxes can then be estimated by maximization or minimization of an objective function using linear inverse modeling algorithms. Furthermore, BiGGR provides functionality to quantify the uncertainty in flux estimates by sampling the constrained multidimensional flux space. As a result, ensembles of possible flux configurations are constructed that agree with measured data within precision limits. BiGGR also features automatic visualization of selected parts of metabolic networks using hypergraphs, with hyperedge widths proportional to estimated flux values. BiGGR supports import and export of models encoded in SBML and is therefore interoperable with different modeling and analysis tools. As an application example, we calculated the flux distribution in healthy human brain using a model of central carbon metabolism. We introduce a new algorithm termed Least-squares with equalities and inequalities Flux Balance Analysis (Lsei-FBA) to predict flux changes from gene expression changes, for instance during disease. Our estimates of brain metabolic flux pattern with Lsei-FBA for Alzheimer's disease agree with independent measurements of cerebral metabolism in patients. This second version of BiGGR is available from Bioconductor.
Fulltext Using GIS and stakeholder involvement to innovate marine mammal bycatch risk assessment in data-limited fisheries

Verutes GM, Johnson AF, Caillat M, Ponnampalam LS, Peter C, Vu L, et al.

PLoS One, 2020;15(8):e0237835.
PMID: 32817725 DOI: 10.1371/journal.pone.0237835

Fisheries bycatch has been identified as the greatest threat to marine mammals worldwide. Characterizing the impacts of bycatch on marine mammals is challenging because it is difficult to both observe and quantify, particularly in small-scale fisheries where data on fishing effort and marine mammal abundance and distribution are often limited. The lack of risk frameworks that can integrate and visualize existing data have hindered the ability to describe and quantify bycatch risk. Here, we describe the design of a new geographic information systems tool built specifically for the analysis of bycatch in small-scale fisheries, called Bycatch Risk Assessment (ByRA). Using marine mammals in Malaysia and Vietnam as a test case, we applied ByRA to assess the risks posed to Irrawaddy dolphins (Orcaella brevirostris) and dugongs (Dugong dugon) by five small-scale fishing gear types (hook and line, nets, longlines, pots and traps, and trawls). ByRA leverages existing data on animal distributions, fisheries effort, and estimates of interaction rates by combining expert knowledge and spatial analyses of existing data to visualize and characterize bycatch risk. By identifying areas of bycatch concern while accounting for uncertainty using graphics, maps and summary tables, we demonstrate the importance of integrating available geospatial data in an accessible format that taps into local knowledge and can be corroborated by and communicated to stakeholders of data-limited fisheries. Our methodological approach aims to meet a critical need of fisheries managers: to identify emergent interaction patterns between fishing gears and marine mammals and support the development of management actions that can lead to sustainable fisheries and mitigate bycatch risk for species of conservation concern.
Fulltext Use of the waxworm Galleria mellonella larvae as an infection model to study Acinetobacter baumannii

Ten KE, Muzahid NH, Rahman S, Tan HS

PLoS One, 2023;18(4):e0283960.
PMID: 37018343 DOI: 10.1371/journal.pone.0283960

Galleria mellonella larvae have been increasingly used in research, including microbial infection studies. They act as suitable preliminary infection models to study host-pathogen interactions due to their advantages, such as the ability to survive at 37°C mimicking human body temperature, their immune system shares similarities with mammalian immune systems, and their short life cycle allowing large-scale studies. Here, we present a protocol for simple rearing and maintenance of G. mellonella without requiring special instruments and specialized training. This allows the continuous supply of healthy G. mellonella for research purposes. Besides, this protocol also provides detailed procedures on the (i) G. mellonella infection assays (killing assay and bacterial burden assay) for virulence studies and (ii) bacterial cell harvesting from infected larvae and RNA extraction for bacterial gene expression studies during infection. Our protocol could not only be used in the studies of A. baumannii virulence but can also be modified according to different bacterial strains.
Fulltext Use of evidence-based practices in pregnancy and childbirth: South East Asia Optimising Reproductive and Child Health in Developing Countries project

SEA-ORCHID Study Group, Laopaiboon M, Lumbiganon P, McDonald SJ, Henderson-Smart DJ, Green S, et al.

PLoS One, 2008 Jul 09;3(7):e2646.
PMID: 18612381 DOI: 10.1371/journal.pone.0002646

BACKGROUND: The burden of mortality and morbidity related to pregnancy and childbirth remains concentrated in developing countries. SEA-ORCHID (South East Asia Optimising Reproductive and Child Health In Developing countries) is evaluating whether a multifaceted intervention to strengthen capacity for research synthesis, evidence-based care and knowledge implementation improves adoption of best clinical practice recommendations leading to better health for mothers and babies. In this study we assessed current practices in perinatal health care in four South East Asian countries and determined whether they were aligned with best practice recommendations.
METHODOLOGY/PRINCIPAL FINDINGS: We completed an audit of 9550 medical records of women and their 9665 infants at nine hospitals; two in each of Indonesia, Malaysia and The Philippines, and three in Thailand between January-December 2005. We compared actual clinical practices with best practice recommendations selected from the Cochrane Library and the World Health Organization Reproductive Health Library. Evidence-based components of the active management of the third stage of labour and appropriately treating eclampsia with magnesium sulphate were universally practiced in all hospitals. Appropriate antibiotic prophylaxis for caesarean section, a beneficial form of care, was practiced in less than 5% of cases in most hospitals. Use of the unnecessary practices of enema in labour ranged from 1% to 61% and rates of episiotomy for vaginal birth ranged from 31% to 95%. Other appropriate practices were commonly performed to varying degrees between countries and also between hospitals within the same country.
CONCLUSIONS/SIGNIFICANCE: Whilst some perinatal health care practices audited were consistent with best available evidence, several were not. We conclude that recording of clinical practices should be an essential step to improve quality of care. Based on these findings, the SEA-ORCHID project team has been developing and implementing interventions aimed at increasing compliance with evidence-based clinical practice recommendations to improve perinatal practice in South East Asia.
Fulltext Use of chronic kidney disease to enhance prediction of cardiovascular risk in those at medium risk

Chia YC, Lim HM, Ching SM

PLoS One, 2015;10(10):e0141344.
PMID: 26496190 DOI: 10.1371/journal.pone.0141344

Based on global cardiovascular (CV) risk assessment for example using the Framingham risk score, it is recommended that those with high risk should be treated and those with low risk should not be treated. The recommendation for those of medium risk is less clear and uncertain. We aimed to determine whether factoring in chronic kidney disease (CKD) will improve CV risk prediction in those with medium risk. This is a 10-year retrospective cohort study of 905 subjects in a primary care clinic setting. Baseline CV risk profile and serum creatinine in 1998 were captured from patients record. Framingham general cardiovascular disease risk score (FRS) for each patient was computed. All cardiovascular disease (CVD) events from 1998-2007 were captured. Overall, patients with CKD had higher FRS risk score (25.9% vs 20%, p = 0.001) and more CVD events (22.3% vs 11.9%, p = 0.002) over a 10-year period compared to patients without CKD. In patients with medium CV risk, there was no significant difference in the FRS score among those with and without CKD (14.4% vs 14.6%, p = 0.84) However, in this same medium risk group, patients with CKD had more CV events compared to those without CKD (26.7% vs 6.6%, p = 0.005). This is in contrast to patients in the low and high risk group where there was no difference in CVD events whether these patients had or did not have CKD. There were more CV events in the Framingham medium risk group when they also had CKD compared those in the same risk group without CKD. Hence factoring in CKD for those with medium risk helps to further stratify and identify those who are actually at greater risk, when treatment may be more likely to be indicated.
Use of a Diagnostic Score to Prioritize Computed Tomographic (CT) Imaging for Patients Suspected of Ischemic Stroke Who May Benefit from Thrombolytic Therapy

Hwong WY, Bots ML, Selvarajah S, Kappelle LJ, Abdul Aziz Z, Sidek NN, et al.

PLoS One, 2016;11(10):e0165330.
PMID: 27768752 DOI: 10.1371/journal.pone.0165330

A shortage of computed tomographic (CT) machines in low and middle income countries often results in delayed CT imaging for patients suspected of a stroke. Yet, time constraint is one of the most important aspects for patients with an ischemic stroke to benefit from thrombolytic therapy. We set out to assess whether application of the Siriraj Stroke Score is able to assist physicians in prioritizing patients with a high probability of having an ischemic stroke for urgent CT imaging.
Fulltext Use of FabV-Triclosan Plasmid Selection System for Efficient Expression and Production of Recombinant Proteins in Escherichia coli

Ali SA, Chew YW, Omar TC, Azman N

PLoS One, 2015;10(12):e0144189.
PMID: 26642325 DOI: 10.1371/journal.pone.0144189

Maintenance of recombinant plasmid vectors in host bacteria relies on the presence of selection antibiotics in the growth media to suppress plasmid -free segregants. However, presence of antibiotic resistance genes and antibiotics themselves is not acceptable in several applications of biotechnology. Previously, we have shown that FabV-Triclosan selection system can be used to select high and medium copy number plasmid vectors in E. coli. Here, we have extended our previous work and demonstrated that expression vectors containing FabV can be used efficiently to express heterologous recombinant proteins in similar or better amounts in E. coli host when compared with expression vectors containing β-lactamase. Use of small amount of non-antibiotic Triclosan as selection agent in growth medium, enhanced plasmid stability, applicability in various culture media, and compatibility with other selection systems for multiple plasmid maintenance are noteworthy features of FabV-Triclosan selection system.

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