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Biomedical subjects

C T Lim

Publications and source records attributed to C T Lim.

At least 19 recordsLinked to original sources

Bead-based microfluidic immunoassays: the next generation.

Microfluidic devices possess many advantages like high throughput, short analysis time, small volume and high sensitivity that fulfill all the important criteria of an immunoassay used for clinical diagnoses, environmental analyses and biochemical studies. These devices can be made from a few different materials, with polymers presently emerging as the most popular choice. Other than being optically clear, non-toxic and cheap, polymers can also be easily fabricated with a variety of techniques. In addition, there are many polymer surface modification methods available to improve the efficiency of these devices. Unfortunately, current microfluidic immunoassays have limited multiplexing capability compared to flow cytometric assays. Flow cytometry employ the use of encoded microbeads in contrast with normal or paramagnetic microbeads applied in current microfluidic devices. The encoded microbead is the key in providing multiplexing capability to the assay by allowing multi-analyte analysis. Using several unique sets of code, different analytes can be detected in a single assay by tracing the identity of individual beads. The same principle could be applied to microfluidic immunoassays in order to retain all the advantages of a fluidic device and significantly improve multiplexing capability.

Animals↗

Characterization of bulk properties of nanofibrous scaffolds from nanomechanical properties of single nanofibers.

This article presents a method of characterizing the macromechanical properties of polymer nanofibrous scaffolds based on the nanomechanical properties obtained for a single nanofiber. Here, we use the example of a poly(L-lactic acid) (PLLA) nanofibrous scaffold that can be used for tissue engineering application. The elastic modulus of a single nanofiber obtained previously from a nanoscale three-point bend test performed using atomic force microscopy was used to model the elastic modulus and elastic collapse stress of the bulk scaffold. Various open-cell solid models are examined for the modeling of bulk scaffold behavior.

Biocompatible Materials↗

Mechanical models for living cells--a review.

As physical entities, living cells possess structural and physical properties that enable them to withstand the physiological environment as well as mechanical stimuli occurring within and outside the body. Any deviation from these properties will not only undermine the physical integrity of the cells, but also their biological functions. As such, a quantitative study in single cell mechanics needs to be conducted. In this review, we will examine some mechanical models that have been developed to characterize mechanical responses of living cells when subjected to both transient and dynamic loads. The mechanical models include the cortical shell-liquid core (or liquid drop) models which are widely applied to suspended cells; the solid model which is generally used for adherent cells; the power-law structural damping model which is more suited for studying the dynamic behavior of adherent cells; and finally, the biphasic model which has been widely used to study musculoskeletal cell mechanics. Based upon these models, future attempts can be made to develop even more detailed and accurate mechanical models of living cells once these three factors are adequately addressed: structural heterogeneity, appropriate constitutive relations for each of the distinct subcellular regions and components, and active forces acting within the cell. More realistic mechanical models of living cells can further contribute towards the study of mechanotransduction in cells.

Animals↗

Coaxial electrospinning of (fluorescein isothiocyanate-conjugated bovine serum albumin)-encapsulated poly(epsilon-caprolactone) nanofibers for sustained release.

As an aim toward developing biologically mimetic and functional nanofiber-based tissue engineering scaffolds, we demonstrated the encapsulation of a model protein, fluorescein isothiocyanate-conjugated bovine serum albumin (fitcBSA), along with a water-soluble polymer, poly(ethylene glycol) (PEG), within the biodegradable poly(epsilon-caprolactone) (PCL) nanofibers using a coaxial electrospinning technique. By variation of the inner flow rates from 0.2 to 0.6 mL/h with a constant outer flow rate of 1.8 mL/h, fitcBSA loadings of 0.85-2.17 mg/g of nanofibrous membranes were prepared. Variation of flow rates also resulted in increases of fiber sizes from ca. 270 nm to 380 nm. The encapsulation of fitcBSA/PEG within PCL was subsequently characterized by laser confocal scanning microscopy, transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS) analysis. In vitro release studies were conducted to evaluate sustained release potential of the core-sheath-structured composite nanofiber PCL-r-fitcBSA/PEG. As a negative control, composite nanofiber PCL/fitcBSA/PEG blend was prepared from a normal electrospinning method. It was found that core-sheath nanofibers PCL-r-fitcBSA/PEG pronouncedly alleviated the initial burst release for higher protein loading and gave better sustainability compared to that of PCL/fitcBSA/PEG nanofibers. The present study would provide a basis for further design and optimization of processing conditions to control the nanostructure of core-sheath composite nanofibers and ultimately achieve desired release kinetics of bioactive proteins (e.g., growth factors) for practical tissue engineering applications.

Animals↗

Synthesis of "cactus" top-decorated aligned carbon nanotubes and their third-order nonlinear optical properties.

We report a new morphology of "cactus" top-decorated aligned carbon nanotubes grown by the PECVD method using pure C2H2 gas. Unlike most previous reports, no additional carrier gas is used for pretreatment. Carbon nanotubes can still grow and maintain the tubular structure underneath the "cactus" tops. It is proposed that the H atoms produced by the dissociation of C2H2 activate the catalyst nanoparticles. Scanning electron microscopy (SEM) shows that the top "cactus" morphology is composed of a large quantity of small nanosheets. Transmission electron microscopy (TEM) reveals the amorphous carbon nature of these "cactus" structures. The formation of these "cactus" structures is possibly due to covalent absorption and reconstruction of carbon atoms on the broken graphite layers of nanotubes produced by the strong ion bombardment under plasma. The third-order optical nonlinearities and nonlinear dynamics are also investigated. The third-order nonlinear susceptibility magnitude /chi(3)/ is found to be 2.2 x 10(-11) esu, and the relaxation process takes place in about 1.8 ps.

Cactaceae↗

Spectrin-level modeling of the cytoskeleton and optical tweezers stretching of the erythrocyte.

We present a three-dimensional computational study of whole-cell equilibrium shape and deformation of human red blood cell (RBC) using spectrin-level energetics. Random network models consisting of degree-2, 3, ..., 9 junction complexes and spectrin links are used to populate spherical and biconcave surfaces and intermediate shapes, and coarse-grained molecular dynamics simulations are then performed with spectrin connectivities fixed. A sphere is first filled with cytosol and gradually deflated while preserving its total surface area, until cytosol volume consistent with the real RBC is reached. The equilibrium shape is determined through energy minimization by assuming that the spectrin tetramer links satisfy the worm-like chain free-energy model. Subsequently, direct stretching by optical tweezers of the initial equilibrium shape is simulated to extract the variation of axial and transverse diameters with the stretch force. At persistence length p = 7.5 nm for the spectrin tetramer molecule and corresponding in-plane shear modulus mu(0) approximately 8.3 microN/m, our models show reasonable agreement with recent experimental measurements on the large deformation of RBC with optical tweezers. We find that the choice of the reference state used for the in-plane elastic energy is critical for determining the equilibrium shape. If a position-independent material reference state such as a full sphere is used in defining the in-plane energy, then the bending modulus kappa needs to be at least a decade larger than the widely accepted value of 2 x 10(-19) J to stabilize the biconcave shape against the cup shape. We demonstrate through detailed computations that this paradox can be avoided by invoking the physical hypothesis that the spectrin network undergoes constant remodeling to always relax the in-plane shear elastic energy to zero at any macroscopic shape, at some slow characteristic timescale. We have devised and implemented a liquefied network structure evolution algorithm that relaxes shear stress everywhere in the network and generates cytoskeleton structures that mimic experimental observations.

Algorithms↗

Connections between single-cell biomechanics and human disease states: gastrointestinal cancer and malaria.

We investigate connections between single-cell mechanical properties and subcellular structural reorganization from biochemical factors in the context of two distinctly different human diseases: gastrointestinal tumor and malaria. Although the cell lineages and the biochemical links to pathogenesis are vastly different in these two cases, we compare and contrast chemomechanical pathways whereby intracellular structural rearrangements lead to global changes in mechanical deformability of the cell. This single-cell biomechanical response, in turn, seems to mediate cell mobility and thereby facilitates disease progression in situations where the elastic modulus increases or decreases due to membrane or cytoskeleton reorganization. We first present new experiments on elastic response and energy dissipation under repeated tensile loading of epithelial pancreatic cancer cells in force- or displacement-control. Energy dissipation from repeated stretching significantly increases and the cell's elastic modulus decreases after treatment of Panc-1 pancreatic cancer cells with sphingosylphosphorylcholine (SPC), a bioactive lipid that influences cancer metastasis. When the cell is treated instead with lysophosphatidic acid, which facilitates actin stress fiber formation, neither energy dissipation nor modulus is noticeably affected. Integrating recent studies with our new observations, we ascribe these trends to possible SPC-induced reorganization primarily of keratin network to perinuclear region of cell; the intermediate filament fraction of the cytoskeleton thus appears to dominate deformability of the epithelial cell. Possible consequences of these results to cell mobility and cancer metastasis are postulated. We then turn attention to progressive changes in mechanical properties of the human red blood cell (RBC) infected with the malaria parasite Plasmodium falciparum. We present, for the first time, continuous force-displacement curves obtained from in-vitro deformation of RBC with optical tweezers for different intracellular developmental stages of parasite. The shear modulus of RBC is found to increase up to 10-fold during parasite development, which is a noticeably greater effect than that from prior estimates. By integrating our new experimental results with published literature on deformability of Plasmodium-harbouring RBC, we examine the biochemical conditions mediating increases or decreases in modulus, and their implications for disease progression. Some general perspectives on connections among structure, single-cell mechanical properties and biological responses associated with pathogenic processes are also provided in the context of the two diseases considered in this work.

Animals↗

Tensile testing of a single ultrafine polymeric fiber.

Due to the difficulty in handling micro and nanoscale fibers and measuring the small load required for deformation, mechanical properties of these fibers have not been widely characterized. In this study, tensile test of a single-strand polycaprolactone electrospun ultrafine fiber was performed using a nano tensile tester. The tested fiber exhibited the characteristic low strength and low modulus but high extensibility at room temperature. The mechanical properties were also found to be dependent on fiber diameter. Fibers with smaller diameter had higher strength but lower ductility due to the higher 'draw ratio' that was applied during the electrospinning process.

Biocompatible Materials↗

Nanoindentation study of human premolars subjected to bleaching agent.

Bleaching of teeth is gaining popularity due to cosmetic reasons. However, the effect it has on teeth is still largely unknown. This paper seeks to evaluate the effect of a bleaching agent, 30% hydrogen peroxide, on the nanomechanical properties of dentin and enamel using the nanoindentation technique. The Young's modulus and hardness obtained from nanoindentation before and after bleaching were compared. Five newly extracted human premolars were used. Nanoindentation was first done on the sliced enamel and dentin regions to determine their mechanical properties. One batch of samples was kept in Hank's balanced salt solution as control while the other was bleached in 30% hydrogen peroxide for 24h. The same number of nanoindentations was then done near the previously indented regions for both the control and bleached samples and the results compared. Using paired sample t-tests with alpha=0.05, it was found that there were no significant differences in both the Young's modulus and hardness of dentin and enamel kept in control. However, the mechanical properties of the bleached dentin were significantly decreased. For intertubular dentin, the mean hardness decreased by 29-55% and the mean Young's modulus decreased by 19-43%. For enamel, the mean hardness decreased by 13-32% while the mean Young's modulus decreased by 18-32%. The exact mechanism by which hydrogen peroxide affects the dentin and enamel has yet to be fully elucidated. However, it is observed to have an undermining effect on the nanomechanical properties of teeth.

Bicuspid↗

Factors affecting nutritional status of Malaysian primary school children.

This paper investigates the nutritional status of a randomly selected cohort of school children and the factors affecting it. This random survey was conducted in the state of Selangor, involving 1,405 primary students (aged 9-10 years from 54 national primary schools). Physical examination was carried out on all the students. Information on the students was also obtained from the parents. Blood samples were taken by using the finger pricking technique. Body mass index (BMI) was used as a measure of physical growth. The students were mainly from urban areas (82.9%). The mean age was 9.71 years and a higher proportion was females (51%). Malays constituted 83.6%, Indians 11.6% and Chinese 4.2% of the study population. The mean weight and height were 32.30 kg and 135.18 cm respectively. The mean BMI was 17.42 kg/m2, with 1.2% of the students underweight, 76.3% normal BMI, 16.3% overweight and 6.3% were obese. Nutritional status was significantly related to blood pressure, history of breast feeding, eating fast food, taking canned/bottled drinks, income and educational level of parents. Significant differences in nutritional status between sexes and locations (rural/urban) were also found. The prevalence of overweight and obese children was of concern. There is thus an urgent need for the School Health Program to periodically monitor the school children's eating habits and physical growth. Appropriate counselling on nutritional intake and physical activities should be given not only to schoolchildren but also to their teachers and parents or caregivers.

Child↗

Effects of nutritional status on academic performance of Malaysian primary school children.

Numerous factors are known to affect the academic performance of students. These include prenatal conditions, birth conditions, postnatal events, nutritional, socio-economic factors and environmental factors. This paper examines the nutritional status and its relationship with academic performance of 9-10 years old primary school children recruited randomly in Selangor, Malaysia. A standard self-administered questionnaire was utilized to obtain pertinent information and a face-to-face interview was also conducted with the parents. Results of the academic performances were extracted from the students' report cards. The intellectual performance was assessed using Raven's Coloured Progressive Matrices. Physical examination was also conducted on these students by doctors. Overall 1,405 students and 1,317 parents responded to the survey. Of these 83.6% were Malays, 11.6% Indians, and 4.2% Chinese. The majority of them (82.9%) were from urban areas. The female: male ratio was 51:49; mean age was 9.71 years. The mean height and weight were 32.3 kg and 135.2 cm respectively. Their mean BMI was 17.42 kg/cm2, with 0.9% underweight, 76.3% normal BMI, 16.3% overweight, and 6.3% obese. Academic performance was significantly correlated with breast feeding, income and educational level of their parents, BMI, and whether they have been taking breakfast. There was a weak correlation between presence of anaemia and intellectual performance. Improving the socio-economic status of the parents will lend a helping hand in the academic performance of the students. Since breast feeding is associated with better academic and intellectual performance it must be emphasized, particularly to expectant mothers in the antenatal clinics.

Child↗

Biophysical approaches for studying the integrity and function of tight junctions.

Cell-cell adhesion is an extremely important phenomenon as it influences several biologically important processes such as inflammation, cell migration, proliferation, differentiation and even cancer metastasis. Furthermore, proteins involved in cell-cell adhesion are also important from the perspective of facilitating better drug delivery across epithelia. The adhesion forces imparted by proteins involved in cell-cell adhesion have been the focus of research for sometime. However, with the advent of nanotechnological techniques such as the atomic force microscopy (AFM), we can now quantitatively probe these adhesion forces not only at the cellular but also molecular level. Here, we review the structure and function of tight junction proteins, highlighting some mechanistic studies performed to quantify the adhesion occurring between these proteins and where possible their association with human diseases. In particular, we will highlight two important experimental techniques, namely the micropipette step pressure technique and the AFM that allow us to quantify these adhesion forces at both the cellular and molecular levels, respectively.

Biophysics↗

Effect of hydrogen peroxide on intertubular dentine.

OBJECTIVES: To evaluate the effect of 30% hydrogen peroxide on the surface changes and nanomechanical properties of intertubular dentine. METHODS: Five freshly extracted human premolars were sectioned and two specimens were obtained from each tooth. Atomic force microscopy (AFM) images of dentine were taken. Baseline hardness and Young's modulus of intertubular dentine were determined using nanoindentation testing and recorded. The specimens were then treated with either 0.5 ml of Hank's balanced salt solution (control group) or 30% hydrogen peroxide at 24 degrees C for 24 h. AFM imaging and determination of hardness and Young's modulus of intertubular dentine were then repeated. The images were compared and statistical analyses of hardness and Young's modulus were carried out using paired sample t-tests. RESULTS: The AFM images showed recession of the intertubular dentine surface post-bleaching. For the control group, there was no significant change in hardness (p=0.124) and Young's modulus (p=0.438) of intertubular dentine. However, a significant decrease in hardness (p=0.002) and Young's modulus (p=0.001) of intertubular dentine was observed for the experimental group. CONCLUSIONS: Exposure to 30% hydrogen peroxide for 24 h caused surface changes to intertubular dentine and significantly decreased the hardness and Young's modulus of intertubular dentine.

Dentin↗

Flexural strength of dental composite restoratives: comparison of biaxial and three-point bending test.

This study compared two test methods used to evaluate the flexural strength of resin-based dental composites. The two test methods evaluated were the three-point bending test4 and the biaxial flexural test. Materials used in this investigation were from the same manufacturer (3M ESPE) and included microfill (A110), minifill (Z100 and Filtek Z250), polyacid modified (F2000), and flowable [Filtek Flowable (FF)] composites. Flexural strength was determined with the use of both test methods after 1 week of conditioning in water at 37 degrees C. Data were analyzed with the use of an ANOVA/Scheffe test and an independent-samples t test at significance level 0.05. Mean flexural strength (n = 7) ranged from 66.61 to 147.21 and 67.27 to 182.81 MPa for three-point bending and ball-on-three-ball biaxial test methods, respectively. In both test methods, Z100 was significantly stronger than all other composites evaluated. In the three-point bending test, flexural strength of Z250 was significantly higher than A110, F2000 and FF, and FF was significantly stronger than A110 and F2000. The biaxial test method arrived at the same conclusions except that there was no significant difference between Z250 and FF. Pearson's correlation revealed a significantly (p < 0.01) positive and good correlation (R2 = 0.72) in flexural strength between the two test methods. Although the biaxial test has the advantage of utilizing small specimens, the low reproducibility of this test method does not support the proposition that it is a more reliable test method when compared to the ISO three-point bending test.

Algorithms↗

Nonlinear elastic and viscoelastic deformation of the human red blood cell with optical tweezers.

Studies of the deformation characteristics of single biological cells can offer insights into the connections among mechanical state, biochemical response and the onset and progression of diseases. Deformation imposed by optical tweezers provides a useful means for the study of single cell mechanics under a variety of well-controlled stress-states. In this paper, we first critically review recent advances in the study of single cell mechanics employing the optical tweezers method, and assess its significance and limitations in comparison to other experimental tools. We then present new experimental and computational results on shape evolution, force-extension curves, elastic properties and viscoelastic response of human red blood cells subjected to large elastic deformation using optical tweezers. Potential applications of the methods examined here to study diseased cells are also briefly addressed.

Elasticity↗

Retrospective review of serologic rubella activity in University Hospital Kuala Lumpur.

A retrospective review of rubella serological results carried out in the Virus Diagnostic Unit, University Hospital Kuala Lumpur (UHKL) from January 1993 to September 1999 showed the presence of rubella infection annually which appeared to increase periodically every two to three years. There was no statistical significant difference in the rubella positive rate between male and female population aged 14 to 48 years. Congenital rubella infections (CRI) occurred in babies delivered in UHKL yearly from 1993 to 1998 with an average incidence rate of 48 per 100,000 deliveries. Peaks of rubella cases appeared to be followed by an increase in incidence of CRI cases 6 to 9 months later. The study showed that only 50.8% clinically diagnosed rubella was confirmed by laboratory finding. This study also showed an increase of rubella activity for the months of July, August and September 1999 and this may herald an increase of CRI cases in the coming millennium.

Adolescent↗