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Fulltext The validity and reliability of motion analysis in patellar tendon reflex assessment

Tham LK, Abu Osman NA, Wan Abas WA, Lim KS

PLoS One, 2013;8(2):e55702.
PMID: 23409022 DOI: 10.1371/journal.pone.0055702

The deep tendon reflex assessments that are essential to the accurate diagnosis of neurological or neuromuscular disorders are conducted subjectively in clinical neurology. Our aim was to assess deep tendon reflexes objectively with a new reflex quantification method.
A novel dengue fever (DF) and dengue haemorrhagic fever (DHF) analysis using artificial neural network (ANN)

Ibrahim F, Taib MN, Abas WA, Guan CC, Sulaiman S

Comput Methods Programs Biomed, 2005 Sep;79(3):273-81.
PMID: 15925426

Dengue fever (DF) is an acute febrile viral disease frequently presented with headache, bone or joint and muscular pains, and rash. A significant percentage of DF patients develop a more severe form of disease, known as dengue haemorrhagic fever (DHF). DHF is the complication of DF. The main pathophysiology of DHF is the development of plasma leakage from the capillary, resulting in haemoconcentration, ascites, and pleural effusion that may lead to shock following defervescence of fever. Therefore, accurate prediction of the day of defervescence of fever is critical for clinician to decide on patient management strategy. To date, no known literature describes of any attempt to predict the day of defervescence of fever in DF patients. This paper describes a non-invasive prediction system for predicting the day of defervescence of fever in dengue patients using artificial neural network. The developed system bases its prediction solely on the clinical symptoms and signs and uses the multilayer feed-forward neural networks (MFNN). The results show that the proposed system is able to predict the day of defervescence in dengue patients with 90% prediction accuracy.
An integrated lateral flow assay for effective DNA amplification and detection at the point of care

Choi JR, Hu J, Gong Y, Feng S, Wan Abas WA, Pingguan-Murphy B, et al.

Analyst, 2016 05 10;141(10):2930-9.
PMID: 27010033 DOI: 10.1039/c5an02532j

Lateral flow assays (LFAs) have been extensively explored in nucleic acid testing (NAT) for medical diagnostics, food safety analysis and environmental monitoring. However, the amount of target nucleic acid in a raw sample is usually too low to be directly detected by LFAs, necessitating the process of amplification. Even though cost-effective paper-based amplification techniques have been introduced, they have always been separately performed from LFAs, hence increasing the risk of reagent loss and cross-contaminations. To date, integrating paper-based nucleic acid amplification into colorimetric LFA in a simple, portable and cost-effective manner has not been introduced. Herein, we developed an integrated LFA with the aid of a specially designed handheld battery-powered system for effective amplification and detection of targets in resource-poor settings. Interestingly, using the integrated paper-based loop-mediated isothermal amplification (LAMP)-LFA, we successfully performed highly sensitive and specific target detection, achieving a detection limit of as low as 3 × 10(3) copies of target DNA, which is comparable to the conventional tube-based LAMP-LFA in an unintegrated format. The device may serve in conjunction with a simple paper-based sample preparation to create a fully integrated paper-based sample-to-answer diagnostic device for point-of-care testing (POCT) in the near future.
Paper-based point-of-care testing for diagnosis of dengue infections

Choi JR, Hu J, Wang S, Yang H, Wan Abas WA, Pingguan-Murphy B, et al.

Crit Rev Biotechnol, 2017 Feb;37(1):100-111.
PMID: 26912259

Dengue endemic is a serious healthcare concern in tropical and subtropical countries. Although well-established laboratory tests can provide early diagnosis of acute dengue infections, access to these tests is limited in developing countries, presenting an urgent need to develop simple, rapid, and robust diagnostic tools. Point-of-care (POC) devices, particularly paper-based POC devices, are typically rapid, cost-effective and user-friendly, and they can be used as diagnostic tools for the prompt diagnosis of dengue at POC settings. Here, we review the importance of rapid dengue diagnosis, current dengue diagnostic methods, and the development of paper-based POC devices for diagnosis of dengue infections at the POC.
Evaluation of postural steadiness in below-knee amputees when wearing different prosthetic feet during various sensory conditions using the Biodex® Stability System

Arifin N, Abu Osman NA, Ali S, Gholizadeh H, Wan Abas WA

Proc Inst Mech Eng H, 2015 Jul;229(7):491-8.
PMID: 26019139 DOI: 10.1177/0954411915587595

In recent years, computerized posturography has become an essential tool in quantitative assessment of postural steadiness in the clinical settings. The purpose of this study was to explore the ability of the Biodex(®) Stability System (BSS) to quantify postural steadiness in below-knee amputees. A convenience sample of 10 below-knee amputees participated in the study. The overall (OSI), anterior-posterior (APSI) and medial-lateral (MLSI) stability indexes as well as the percentage of time spent in left and right quadrants and four concentric zones were measured under altered sensory conditions while standing with solid ankle cushion heel (SACH), single-axis (SA) and energy storage and release (ESAR) feet. Significant difference was found between sensory conditions in SACH and ESAR feet for OSI (SACH, p = 0.002; ESAR, p = 0.005), APSI (SACH, p = 0.036; ESAR, p = 0.003) and MLSI (SACH, p = 0.008; ESAR, p = 0.05) stability indexes. The percentage of time spent in Zone A (0°-5°) was significantly greater than the other three concentric zones (p < 0.01). The loading time percentage on their intact limb (80%-94%) was significantly longer than the amputated limb (20%-6%) in all conditions for all three prosthetic feet. Below-knee amputees showed compromised postural steadiness when visual, proprioceptive or vestibular sensory input was altered. The findings highlight that the characteristics of postural stability in amputees can be clinically assessed by utilizing the outcomes produced by the BSS.
Paper-based sample-to-answer molecular diagnostic platform for point-of-care diagnostics

Choi JR, Tang R, Wang S, Wan Abas WA, Pingguan-Murphy B, Xu F

Biosens Bioelectron, 2015 Dec 15;74:427-39.
PMID: 26164488 DOI: 10.1016/j.bios.2015.06.065

Nucleic acid testing (NAT), as a molecular diagnostic technique, including nucleic acid extraction, amplification and detection, plays a fundamental role in medical diagnosis for timely medical treatment. However, current NAT technologies require relatively high-end instrumentation, skilled personnel, and are time-consuming. These drawbacks mean conventional NAT becomes impractical in many resource-limited disease-endemic settings, leading to an urgent need to develop a fast and portable NAT diagnostic tool. Paper-based devices are typically robust, cost-effective and user-friendly, holding a great potential for NAT at the point of care. In view of the escalating demand for the low cost diagnostic devices, we highlight the beneficial use of paper as a platform for NAT, the current state of its development, and the existing challenges preventing its widespread use. We suggest a strategy involving integrating all three steps of NAT into one single paper-based sample-to-answer diagnostic device for rapid medical diagnostics in the near future.
Cryopreservation of Human Mesenchymal Stem Cells for Clinical Applications: Current Methods and Challenges

Yong KW, Wan Safwani WK, Xu F, Wan Abas WA, Choi JR, Pingguan-Murphy B

Biopreserv Biobank, 2015 Aug;13(4):231-9.
PMID: 26280501 DOI: 10.1089/bio.2014.0104

Mesenchymal stem cells (MSCs) hold many advantages over embryonic stem cells (ESCs) and other somatic cells in clinical applications. MSCs are multipotent cells with strong immunosuppressive properties. They can be harvested from various locations in the human body (e.g., bone marrow and adipose tissues). Cryopreservation represents an efficient method for the preservation and pooling of MSCs, to obtain the cell counts required for clinical applications, such as cell-based therapies and regenerative medicine. Upon cryopreservation, it is important to preserve MSCs functional properties including immunomodulatory properties and multilineage differentiation ability. Further, a biosafety evaluation of cryopreserved MSCs is essential prior to their clinical applications. However, the existing cryopreservation methods for MSCs are associated with notable limitations, leading to a need for new or improved methods to be established for a more efficient application of cryopreserved MSCs in stem cell-based therapies. We review the important parameters for cryopreservation of MSCs and the existing cryopreservation methods for MSCs. Further, we also discuss the challenges to be addressed in order to preserve MSCs effectively for clinical applications.
Re: Re: "Quantitative and Qualitative Comparison of a New Prosthetic Suspension System with Two Existing Suspension Systems for Lower Limb Amputees"

Eshraghi A, Osman NA, Karimi MT, Gholizadeh H, Ali S, Wan Abas WA

Am J Phys Med Rehabil, 2015 Jul;94(7):e60.
PMID: 25888670 DOI: 10.1097/PHM.0000000000000304
Investigation to predict patellar tendon reflex using motion analysis technique

Tham LK, Osman NA, Lim KS, Pingguan-Murphy B, Abas WA, Zain NM

Med Eng Phys, 2011 May;33(4):407-10.
PMID: 21146440 DOI: 10.1016/j.medengphy.2010.11.002

The investigation of patellar tendon reflex involves development of a reflex hammer holder, kinematic data collection and analysis of patellar reflex responses using motion analysis techniques. The main aim of this research is to explore alternative means of assessing reflexes as a part of routine clinical diagnosis. The motion analysis system was applied to provide quantitative data which is a more objective measure of the patellar tendon reflex. Kinematic data was collected from 28 males and 22 females whilst subjected to a knee jerk test. Further analysis of kinematic data was performed to predict relationships which might affect the patellar tendon reflex. All subjects were seated on a high stool with their legs hanging freely within the capture volume of the motion analysis system. Knee jerk tests were applied to all subjects, on both sides of the leg, by eliciting hypo, hyper, and normal reflexes. An additional reinforcement technique called the Jendrassik manoeuvre was also performed under the same conditions to elicit a normal patellar tendon reflex. The comparison of reflex response between genders showed that female subjects generally had a greater response compared to males. However, the difference in reflex response between the left leg and the right leg was not significant. Tapping strength to elicit a hyper-reflex produced greater knee-jerk compared to the normal clinical tapping strength. All results were in agreement with clinical findings and results found by some early researchers.
In vitro comparative study of white and dark polycaprolactone trifumarate in situ cross-linkable scaffolds seeded with rat bone marrow stromal cells

Muhammad KB, Abas WA, Kim KH, Pingguan-Murphy B, Zain NM, Akram H

Clinics (Sao Paulo), 2012;67(6):629-38.
PMID: 22760903

OBJECTIVE: Dark poly(caprolactone) trifumarate is a successful candidate for use as a bone tissue engineering scaffold. Recently, a white polymeric scaffold was developed that shows a shorter synthesis time and is more convenient for tissue-staining work. This is an in vitro comparative study of both the white and dark scaffolds.
METHODS: Both white and dark poly(caprolactone) trifumarate macromers were characterized via Fourier transform infrared spectroscopy before being chemically cross-linked and molded into disc-shaped scaffolds. Biodegradability was assessed by percentage weight loss on days 7, 14, 28, 42 and 56 (n = 5) after immersion in 10% serum-supplemented medium or distilled water. Static cell seeding was employed in which isolated and characterized rat bone marrow stromal cells were seeded directly onto the scaffold surface. Seeded scaffolds were subjected to a series of biochemical assays and scanning electron microscopy at specified time intervals for up to 28 days of incubation.
RESULTS: The degradation of the white scaffold was significantly lower compared with the dark scaffold but was within the acceptable time range for bone-healing processes. The deoxyribonucleic acid and collagen contents increased up to day 28 with no significant difference between the two scaffolds, but the glycosaminoglycan content was slightly higher in the white scaffold throughout 14 days of incubation. Scanning electron microscopy at day 1 [corrected] revealed cellular growth and attachment.
CONCLUSIONS: There was no cell growth advantage between the two forms, but the white scaffold had a slower biodegradability rate, suggesting that the newly synthesized poly(caprolactone) trifumarate is more suitable for use as a bone tissue engineering scaffold.
Impact of low oxygen tension on stemness, proliferation and differentiation potential of human adipose-derived stem cells

Choi JR, Pingguan-Murphy B, Wan Abas WA, Noor Azmi MA, Omar SZ, Chua KH, et al.

Biochem Biophys Res Commun, 2014 May 30;448(2):218-24.
PMID: 24785372 DOI: 10.1016/j.bbrc.2014.04.096

Adipose-derived stem cells (ASCs) have been found adapted to a specific niche with low oxygen tension (hypoxia) in the body. As an important component of this niche, oxygen tension has been known to play a critical role in the maintenance of stem cell characteristics. However, the effect of O2 tension on their functional properties has not been well determined. In this study, we investigated the effects of O2 tension on ASCs stemness, differentiation and proliferation ability. Human ASCs were cultured under normoxia (21% O2) and hypoxia (2% O2). We found that hypoxia increased ASC stemness marker expression and proliferation rate without altering their morphology and surface markers. Low oxygen tension further enhances the chondrogenic differentiation ability, but reduces both adipogenic and osteogenic differentiation potential. These results might be correlated with the increased expression of HIF-1α under hypoxia. Taken together, we suggest that growing ASCs under 2% O2 tension may be important in expanding ASCs effectively while maintaining their functional properties for clinical therapy, particularly for the treatment of cartilage defects.
Transtibial prosthetic suspension: less pistoning versus easy donning and doffing

Gholizadeh H, Abu Osman NA, Eshraghi A, Ali S, Sævarsson SK, Wan Abas WA, et al.

J Rehabil Res Dev, 2012;49(9):1321-30.
PMID: 23408214

Poor suspension increases slippage of the residual limb inside the socket during ambulation. The main purpose of this article is to evaluate the pistoning at the prosthetic liner-socket interface during gait and assess patients' satisfaction with two different liners. Two prostheses with seal-in and locking liners were fabricated for each of the 10 subjects with transtibial amputation. The Vicon motion system was used to measure the pistoning during gait. The subjects were also asked to complete a Prosthesis Evaluation Questionnaire. The results revealed higher pistoning inside the socket during gait with the locking liner than with the seal-in liner (p < 0.05). The overall satisfaction with the locking liner was higher (p < 0.05) because of the relative ease with which the patients could don and doff the device. As such, pistoning may not be the main factor that determines patients' overall satisfaction with the prosthesis and other factors may also contribute to comfort and satisfaction with prostheses. The article also verifies the feasibility of the Vicon motion system for measuring pistoning during gait.
The effects of hypoxia and serum-free conditions on the stemness properties of human adipose-derived stem cells

Wan Safwani WK, Wong CW, Yong KW, Choi JR, Mat Adenan NA, Omar SZ, et al.

Cytotechnology, 2016 Oct;68(5):1859-72.
PMID: 26728363 DOI: 10.1007/s10616-015-9939-9

The need to have a better and safer culture condition for expansion of human mesenchymal stem cells (MSCs) is crucial particularly to prevent infection and immune rejection. This is normally associated with the use of animal-based serum in the culture media for cell expansion. The aim of this study is to investigate alternative culture conditions which may provide better and safer environment for cell growth. In the present study, human adipose-derived stem cells (ASCs) at passage 3 were subjected to treatment in 4 conditions: (1) 21 % O2 with fetal bovine serum (FBS), (2) 21 % O2 without FBS, (3) 2 % O2 with FBS and (4) 2 % O2 without FBS followed by subsequent analysis of their phenotype, viability and functionality. We observed that ASCs cultured in all conditions present no significant phenotypic changes. It was found that ASCs cultured in 2 % O2 without serum showed an increase in viability and growth to a certain extent when compared to those cultured in 21 % O2 without serum. However, ASCs cultured in 2 % O2 without serum displayed a relatively low adipogenic and osteogenic potential. On the other hand, interestingly, there was a positive enhancement in chondrogenic differentiation of ASCs cultured in 21 % O2 without serum. Our findings suggest that different culture conditions may be suitable for different indications. In summary, ASCs cultured in serum-free condition can still survive, proliferate and undergo subsequent adipogenic, osteogenic and chondrogenic differentiation. Therefore, FBS is feasible to be excluded for culture of ASCs, which avoids clinical complications.
Fulltext In situ normoxia enhances survival and proliferation rate of human adipose tissue-derived stromal cells without increasing the risk of tumourigenesis

Choi JR, Pingguan-Murphy B, Wan Abas WA, Yong KW, Poon CT, Noor Azmi MA, et al.

PLoS One, 2015;10(1):e0115034.
PMID: 25615717 DOI: 10.1371/journal.pone.0115034

Adipose tissue-derived stromal cells (ASCs) natively reside in a relatively low-oxygen tension (i.e., hypoxic) microenvironment in human body. Low oxygen tension (i.e., in situ normoxia), has been known to enhance the growth and survival rate of ASCs, which, however, may lead to the risk of tumourigenesis. Here, we investigated the tumourigenic potential of ASCs under their physiological condition to ensure their safe use in regenerative therapy. Human ASCs isolated from subcutaneous fat were cultured in atmospheric O2 concentration (21% O2) or in situ normoxia (2% O2). We found that ASCs retained their surface markers, tri-lineage differentiation potential, and self-renewal properties under in situ normoxia without altering their morphology. In situ normoxia displayed a higher proliferation and viability of ASCs with less DNA damage as compared to atmospheric O2 concentration. Moreover, low oxygen tension significantly up-regulated VEGF and bFGF mRNA expression and protein secretion while reducing the expression level of tumour suppressor genes p16, p21, p53, and pRb. However, there were no significant differences in ASCs telomere length and their relative telomerase activity when cultured at different oxygen concentrations. Collectively, even with high proliferation and survival rate, ASCs have a low tendency of developing tumour under in situ normoxia. These results suggest 2% O2 as an ideal culture condition for expanding ASCs efficiently while maintaining their characteristics.
An integrated paper-based sample-to-answer biosensor for nucleic acid testing at the point of care

Choi JR, Hu J, Tang R, Gong Y, Feng S, Ren H, et al.

Lab Chip, 2016 Feb 7;16(3):611-21.
PMID: 26759062 DOI: 10.1039/c5lc01388g

With advances in point-of-care testing (POCT), lateral flow assays (LFAs) have been explored for nucleic acid detection. However, biological samples generally contain complex compositions and low amounts of target nucleic acids, and currently require laborious off-chip nucleic acid extraction and amplification processes (e.g., tube-based extraction and polymerase chain reaction (PCR)) prior to detection. To the best of our knowledge, even though the integration of DNA extraction and amplification into a paper-based biosensor has been reported, a combination of LFA with the aforementioned steps for simple colorimetric readout has not yet been demonstrated. Here, we demonstrate for the first time an integrated paper-based biosensor incorporating nucleic acid extraction, amplification and visual detection or quantification using a smartphone. A handheld battery-powered heating device was specially developed for nucleic acid amplification in POC settings, which is coupled with this simple assay for rapid target detection. The biosensor can successfully detect Escherichia coli (as a model analyte) in spiked drinking water, milk, blood, and spinach with a detection limit of as low as 10-1000 CFU mL(-1), and Streptococcus pneumonia in clinical blood samples, highlighting its potential use in medical diagnostics, food safety analysis and environmental monitoring. As compared to the lengthy conventional assay, which requires more than 5 hours for the entire sample-to-answer process, it takes about 1 hour for our integrated biosensor. The integrated biosensor holds great potential for detection of various target analytes for wide applications in the near future.
Fulltext Phenotypic and functional characterization of long-term cryopreserved human adipose-derived stem cells

Yong KW, Pingguan-Murphy B, Xu F, Abas WA, Choi JR, Omar SZ, et al.

Sci Rep, 2015;5:9596.
PMID: 25872464 DOI: 10.1038/srep09596

Cryopreservation represents an effective technique to maintain the functional properties of human adipose-derived stem cells (ASCs) and allows pooling of cells via long-term storage for clinical applications, e.g., cell-based therapies. It is crucial to reduce freezing injury during the cryopreservation process by loading the ASCs with the optimum concentration of suitable cryoprotective agents (CPAs). In this study, human ASCs were preserved for 3 months in different combinations of CPAs, including 1) 0.25 M trehalose; 2) 5% dimethylsulfoxide (DMSO); 3) 10% DMSO; 4) 5% DMSO + 20% fetal bovine serum (FBS); 5) 10% DMSO + 20% FBS; 6) 10% DMSO + 90% FBS. Interestingly, even with a reduction of DMSO to 5% and without FBS, cryopreserved ASCs maintained high cell viability comparable with standard cryomedium (10% DMSO + 90% FBS), with normal cell phenotype and proliferation rate. Cryopreserved ASCs also maintained their differentiation capability (e.g., to adipocytes, osteocytes and chondrocytes) and showed an enhanced expression level of stemness markers (e.g., NANOG, OCT-4, SOX-2 and REX-1). Our findings suggest that 5% DMSO without FBS may be an ideal CPA for an efficient long-term cryopreservation of human ASCs. These results aid in establishing standardized xeno-free long-term cryopreservation of human ASCs for clinical applications.
Fulltext Paracrine Effects of Adipose-Derived Stem Cells on Matrix Stiffness-Induced Cardiac Myofibroblast Differentiation via Angiotensin II Type 1 Receptor and Smad7

Yong KW, Li Y, Liu F, Bin Gao, Lu TJ, Wan Abas WA, et al.

Sci Rep, 2016 10 05;6:33067.
PMID: 27703175 DOI: 10.1038/srep33067

Human mesenchymal stem cells (hMSCs) hold great promise in cardiac fibrosis therapy, due to their potential ability of inhibiting cardiac myofibroblast differentiation (a hallmark of cardiac fibrosis). However, the mechanism involved in their effects remains elusive. To explore this, it is necessary to develop an in vitro cardiac fibrosis model that incorporates pore size and native tissue-mimicking matrix stiffness, which may regulate cardiac myofibroblast differentiation. In the present study, collagen coated polyacrylamide hydrogel substrates were fabricated, in which the pore size was adjusted without altering the matrix stiffness. Stiffness is shown to regulate cardiac myofibroblast differentiation independently of pore size. Substrate at a stiffness of 30 kPa, which mimics the stiffness of native fibrotic cardiac tissue, was found to induce cardiac myofibroblast differentiation to create in vitro cardiac fibrosis model. Conditioned medium of hMSCs was applied to the model to determine its role and inhibitory mechanism on cardiac myofibroblast differentiation. It was found that hMSCs secrete hepatocyte growth factor (HGF) to inhibit cardiac myofibroblast differentiation via downregulation of angiotensin II type 1 receptor (AT1R) and upregulation of Smad7. These findings would aid in establishment of the therapeutic use of hMSCs in cardiac fibrosis therapy in future.

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