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  1. Fulltext Dielectrophoretic manipulation and separation of microparticles using microarray dot electrodes
    Yafouz B, Kadri NA, Ibrahim F
    Sensors (Basel), 2014;14(4):6356-69.
    PMID: 24705632 DOI: 10.3390/s140406356
    This paper introduces a dielectrophoretic system for the manipulation and separation of microparticles. The system is composed of five layers and utilizes microarray dot electrodes. We validated our system by conducting size-dependent manipulation and separation experiments on 1, 5 and 15 μm polystyrene particles. Our findings confirm the capability of the proposed device to rapidly and efficiently manipulate and separate microparticles of various dimensions, utilizing positive and negative dielectrophoresis (DEP) effects. Larger size particles were repelled and concentrated in the center of the dot by negative DEP, while the smaller sizes were attracted and collected by the edge of the dot by positive DEP.
    Matched MeSH terms: Electrophoresis/methods*
  2. Fulltext Spectroscopic (UV/VIS, Raman) and Electrophoresis Study of Cytosine-Guanine Oligonucleotide DNA Influenced by Magnetic Field
    Banihashemian SM, Periasamy V, Boon Tong G, Abdul Rahman S
    PLoS One, 2016;11(3):e0149488.
    PMID: 26999445 DOI: 10.1371/journal.pone.0149488
    Studying the effect of a magnetic field on oligonucleotide DNA can provide a novel DNA manipulation technique for potential application in bioengineering and medicine. In this work, the optical and electrochemical response of a 100 bases oligonucleotides DNA, cytosine-guanine (CG100), is investigated via exposure to different magnetic fields (250, 500, 750, and 1000 mT). As a result of the optical response of CG100 to the magnetic field, the ultra-violet-visible spectrum indicated a slight variation in the band gap of CG100 of about 0.3 eV. Raman spectroscopy showed a significant deviation in hydrogen and phosphate bonds' vibration after exposure to the magnetic field. Oligonucleotide DNA mobility was investigated in the external electric field using the gel electrophoresis technique, which revealed a small decrease in the migration of CG100 after exposure to the magnetic field.
    Matched MeSH terms: Electrophoresis/methods*
  3. Fulltext Screening for intermediate and severe forms of thalassaemia in discarded red blood cells: optimization and feasibility
    George E, Lai MI, Teh LK, Ramasamy R, Goh EH, Asokan K, et al.
    Med J Malaysia, 2011 Dec;66(5):429-34.
    PMID: 22390095 MyJurnal
    Detection and quantification of Hb subtypes of human blood is integral to presumptive identification of thalassaemias. It has been used in neonatal screening of thalassaemia and Hb variants. The use of discarded red blood cells following processing of the cord blood for stem cells provides readily available diagnostic material for thalassaemia screening. In this study, we determined the range of Hb subtypes in 195 consecutive cord blood samples collected for cord blood banking. The 'cord blood samples' analysed were those of the remaining red blood cells after the cord blood was processed for stem cell storage. Quantification of Hb subtypes by high performance liquid chromatography (HPLC) was done on BioRad Variant II Hb testing system. Only 73 (36.5%) of the samples could be analyzed neat without dilution. With a 1:300 dilution with wash solution the acceptable area as recommended by the manufacturer for reading of a C-gram within the 1 to 3 million ranges were achieved in all. Eighteen (9%) 12 showed classical Hb Barts (y4) prerun peaks were confirmed by Sebia Hydrasys automated Hb gel electrophoresis and quantified by Sebia Capillarys 2 capillary electrophoresis. Only 1 (0.5%) was presumptively identified with HbH disease. Due to the limited number of samples no beta-thalassaemia major, Hb E beta-thalassaemia and Hb Barts hydrops fetalis were found. The HPLC assay was possible at a cost US$ 5 per sample and a turnover time of 10 samples per hour without technical difficulties. This study reports an effective and valuable protocol for thalassaemia screening in red blood cells which would otherwise be discarded during cord blood processing. Cord blood with severe and intermediate forms of thalassaemia can be preselected and not stored.
    Matched MeSH terms: Electrophoresis/methods
  4. Fulltext Separation of sulfated urinary glycosaminoglycans by high-resolution electrophoresis for isotyping of mucopolysaccharidoses in Malaysia
    Nor A, Zabedah MY, Norsiah MD, Ngu LH, Suhaila AR
    Malays J Pathol, 2010 Jun;32(1):35-42.
    PMID: 20614724
    Mucopolysaccharidoses (MPS) are a group of inherited disorders caused by the deficiency of specific lysosomal enzymes involved in glycosaminoglycans (GAGs) degradation. Currently, there are 11 enzyme deficiencies resulting in seven distinct MPS clinical syndromes and their subtypes. Different MPS syndromes cannot be clearly distinguished clinically due to overlapping signs and symptoms. Measurement of GAGs content in urine and separation of GAGs using high-resolution electrophoresis (HRE) are very useful initial screening tests for isotyping of MPS before specific enzyme diagnostics. In this study, we measured total urinary GAGs by a method using dimethylmethylene blue (DMB), and followed by isolation and separation of GAGs using high resolution electrophoresis (HRE) technique. Of 760 urine samples analyzed, 40 have abnormal GAGs HRE patterns. Thirty-five of these 40 cases have elevated urinary GAGs levels as well. These abnormal HRE patterns could be classified into 4 patterns: Pattern A (elevated DS and HS; suggestive of MPS I, II or VII; 16 cases), Pattern B (elevated HS and CS; suggestive of MPS III; 17 cases), and Pattern C (elevated KS and CS; suggestive of MPS IV, 5 cases), and Pattern D (elevated DS; suggestive of MPS VI; 2 cases). Based on the GAGs HRE pattern and a few discriminating clinical signs, we performed selective enzymatic investigation in 16 cases. In all except one case with MPS VII, the enzymatic diagnosis correlated well with the provisional MPS type as suggested by the abnormal HRE pattern. Our results showed that GAGs HRE is a useful, inexpensive and practical first-line screening test when MPS is suspected clinically, and it provides an important guide to further enzymatic studies on a selective basis.
    Matched MeSH terms: Electrophoresis/methods*
  5. Modeling the relationship between the main emulsion components and stability, viscosity, fluid behavior, zeta-potential, and electrophoretic mobility of orange beverage emulsion using response surface methodology
    Mirhosseini H, Tan CP, Hamid NS, Yusof S
    J Agric Food Chem, 2007 Sep 19;55(19):7659-66.
    PMID: 17708646
    The possible relationships between the main emulsion components (namely, Arabic gum, xanthan gum, and orange oil) and the physicochemical properties of orange beverage emulsion were evaluated by using response surface methodology. The physicochemical emulsion property variables considered as response variables were emulsion stability, viscosity, fluid behavior, zeta-potential, and electrophoretic mobility. The independent variables had the most and least significant ( p < 0.05) effect on viscosity and zeta-potential, respectively. The quadratic effect of orange oil and Arabic gum, the interaction effect of Arabic gum and xanthan gum, and the main effect of Arabic gum were the most significant ( p < 0.05) effects on turbidity loss rate, viscosity, viscosity ratio, and mobility, respectively. The main effect of Arabic gum was found to be significant ( p < 0.05) in all response variables except for turbidity loss rate. The nonlinear regression equations were significantly ( p < 0.05) fitted for all response variables with high R (2) values (>0.86), which had no indication of lack of fit. The results indicated that a combined level of 10.78% (w/w) Arabic gum, 0.56% (w/w) xanthan gum, and 15.27% (w/w) orange oil was predicted to provide the overall optimum region in terms of physicochemical properties studied. No significant ( p > 0.05) difference between the experimental and the predicted values confirmed the adequacy of response surface equations.
    Matched MeSH terms: Electrophoresis/methods
  6. Fulltext An alternate method for DNA and RNA extraction from clotted blood
    Zakaria Z, Umi SH, Mokhtar SS, Mokhtar U, Zaiharina MZ, Aziz AT, et al.
    Genet. Mol. Res., 2013;12(1):302-11.
    PMID: 23408417 DOI: 10.4238/2013.February.4.4
    We developed an alternative method to extract DNA and RNA from clotted blood for genomic and molecular investigations. A combination of the TRIzol method and the QIAamp spin column were used to extract RNA from frozen clotted blood. Clotted blood was sonicated and then the QIAamp DNA Blood Mini Kit was used for DNA extraction. Extracted DNA and RNA were adequate for gene expression analysis and copy number variation (CNV) genotyping, respectively. The purity of the extracted RNA and DNA was in the range of 1.8-2.0, determined by absorbance ratios of A(260):A(280). Good DNA and RNA integrity were confirmed using gel electrophoresis and automated electrophoresis. The extracted DNA was suitable for qPCR and microarrays for CNV genotyping, while the extracted RNA was adequate for gene analysis using RT-qPCR.
    Matched MeSH terms: Electrophoresis/methods
  7. Continuous separation of colloidal particles using dielectrophoresis
    Yunus NA, Nili H, Green NG
    Electrophoresis, 2013 Apr;34(7):969-78.
    PMID: 23436439 DOI: 10.1002/elps.201200466
    Dielectrophoresis is the movement of particles in nonuniform electric fields and has been of interest for application to manipulation and separation at and below the microscale. This technique has the advantages of being noninvasive, nondestructive, and noncontact, with the movement of particle achieved by means of electric fields generated by miniaturized electrodes and microfluidic systems. Although the majority of applications have been above the microscale, there is increasing interest in application to colloidal particles around a micron and smaller. This paper begins with a review of colloidal and nanoscale dielectrophoresis with specific attention paid to separation applications. An innovative design of integrated microelectrode array and its application to flow-through, continuous separation of colloidal particles is then presented. The details of the angled chevron microelectrode array and the test microfluidic system are then discussed. The variation in device operation with applied signal voltage is presented and discussed in terms of separation efficiency, demonstrating 99.9% separation of a mixture of colloidal latex spheres.
    Matched MeSH terms: Electrophoresis/methods*
  8. In-Transit Electroextraction of Small-Molecule Pharmaceuticals from Blood
    Nanthasurasak P, See HH, Zhang M, Guijt RM, Breadmore MC
    Angew Chem Int Ed Engl, 2019 03 18;58(12):3790-3794.
    PMID: 30614157 DOI: 10.1002/anie.201812077
    An electrokinetic platform was developed for extracting small-molecule pharmaceuticals from a dried blood spot. Through the exclusion of liquid reagents and use of low field strength (6 V cm-1 ), the electroextraction of a drug from a dried blood spot, deposited on a polymer inclusion membrane (PIM), could be realised while in transit in the mail. In transit sample preparation provides a potential solution to in situ sample degradation and may accelerate the workflow upon arrival of a patient sample at the analytical facility. The electroextraction method was enabled through our discovery of the use of 15-20 μm thin PIMs as electrophoretic separation medium in absence of liquid reagents. Here, a PIM consisting of cellulose triacetate as polymer base, 2-nitrophenyl octyl ether as plasticizer and 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide as carrier was used. The PIM, was packaged with two 12 V batteries to supply the separation voltage. A blood spot containing berberine chloride was deposited and dried before the applying the separation potential, allowing for the electroextraction while the packaged device was shipped in internal mail. Upon arrival in the analytical laboratory, the PIM was analysed using a fluorescence microscope with photon multiplier tube, quantifying the berberine extracted away from the sample matrix. This platform represents a new opportunity for processing clinical samples during transport to the laboratory, saving time and manual handling to accelerate the time to result.
    Matched MeSH terms: Electrophoresis/methods*
  9. Fulltext Dielectrophoresis of Amyloid-Beta Proteins as a Microfluidic Template for Alzheimer's Research
    Al-Ahdal SA, Ahmad Kayani AB, Md Ali MA, Chan JY, Ali T, Adnan N, et al.
    Int J Mol Sci, 2019 Jul 23;20(14).
    PMID: 31340481 DOI: 10.3390/ijms20143595
    We employed dielectrophoresis to a yeast cell suspension containing amyloid-beta proteins (Aβ) in a microfluidic environment. The Aβ was separated from the cells and characterized using the gradual dissolution of Aβ as a function of the applied dielectrophoretic parameters. We established the gradual dissolution of Aβ under specific dielectrophoretic parameters. Further, Aβ in the fibril form at the tip of the electrode dissolved at high frequency. This was perhaps due to the conductivity of the suspending medium changing according to the frequency, which resulted in a higher temperature at the tips of the electrodes, and consequently in the breakdown of the hydrogen bonds. However, those shaped as spheroidal monomers experienced a delay in the Aβ fibril transformation process. Yeast cells exposed to relatively low temperatures at the base of the electrode did not experience a positive or negative change in viability. The DEP microfluidic platform incorporating the integrated microtip electrode array was able to selectively manipulate the yeast cells and dissolve the Aβ to a controlled extent. We demonstrate suitable dielectrophoretic parameters to induce such manipulation, which is highly relevant for Aβ-related colloidal microfluidic research and could be applied to Alzheimer's research in the future.
    Matched MeSH terms: Electrophoresis/methods*
  10. Serum free light chains: diagnostic and prognostic value in multiple myeloma
    Sthaneshwar P, Nadarajan V, Maniam JA, Nordin N, Gin Gin G
    Clin Chem Lab Med, 2009;47(9):1101-7.
    PMID: 19728852 DOI: 10.1515/CCLM.2009.260
    Measurement of serum free light chains (FLCs) has recently become available for the diagnosis and monitoring of patients with plasma cell dyscrasias. The aim of this study was to investigate the performance of the serum FLC assay as a tumour marker by comparing FLC concentrations with serum protein electrophoresis (PE) results in the diagnosis of multiple myeloma (MM). In addition, we also evaluated the prognostic value of the baseline serum FLC ratio in patients with MM.
    Matched MeSH terms: Blood Protein Electrophoresis/methods*
  11. Dielectrophoretic deformation of breast cancer cells for lab on a chip applications
    Chan JY, Ahmad Kayani AB, Md Ali MA, Kok CK, Ramdzan Buyong M, Hoe SLL, et al.
    Electrophoresis, 2019 10;40(20):2728-2735.
    PMID: 31219180 DOI: 10.1002/elps.201800442
    This paper presents the development and experimental analysis of a curved microelectrode platform for the DEP deformation of breast cancer cells (MDA-MB-231). The platform is composed of arrays of curved DEP microelectrodes which are patterned onto a glass slide and samples containing MDA-MB-231 cells are pipetted onto the platform's surface. Finite element method is utilised to characterise the electric field gradient and DEP field. The performance of the system is assessed with MDA-MB-231 cells in a low conductivity 1% DMEM suspending medium. We applied sinusoidal wave AC potential at peak to peak voltages of 2, 5, and 10 Vpp at both 10 kHz and 50 MHz. We observed cell blebbing and cell shrinkage and analyzed the percentage of shrinkage of the cells. The experiments demonstrated higher percentage of cell shrinkage when cells are exposed to higher frequency and peak to peak voltage electric field.
    Matched MeSH terms: Electrophoresis/methods
  12. Fulltext Performance comparison of EasyFix G26 and HYDRASYS 2 SCAN for the detection of serum monoclonal proteins
    Amin Nordin FD, Mohd Khalid MKN, Abdul Aziz SM, Mohamad Bakri NA, Ahmad Ridzuan SN, Abdul Jalil J, et al.
    J Clin Lab Anal, 2020 Jun;34(6):e23254.
    PMID: 32141626 DOI: 10.1002/jcla.23254
    BACKGROUND: Serum protein electrophoresis (SPE) is a widely used laboratory technique to diagnose patients with multiple myeloma (MM) and other disorders related to serum protein. In patients with MM, abnormal monoclonal protein can be detected by SPE and further characterized using immunofixation electrophoresis (IFE). There are several semi-automated agarose gel-based systems available commercially for SPE and IFE. In this study, we sought to evaluate the analytical performance of fully automated EasyFix G26 (EFG26) and semi-automated HYDRASYS 2 SCAN (H2SCAN) for both SPE and IFE.

    METHODS: Both instruments were operated according to manufacturer's instructions. Samples used include a commercially available normal control serum (NCS) and patients' specimens. The following were evaluated: precision and comparison studies for SPE, and reproducibility and comparison studies for IFE. Statistical analyses were performed using Microsoft Excel.

    RESULTS: For SPE repeatability study, our results showed that EFG26 has higher coefficient of variation (%CV) compared with H2SCAN for both samples except for monoclonal component with %CV of 0.97% and 1.18%, respectively. Similar results were obtained for SPE reproducibility study except for alpha-1 (4.16%) and beta (3.13%) fractions for NCS, and beta fractions (5.36%) for monoclonal sample. Subsequently, reproducibility for IFE was 100% for both instruments. Values for correlation coefficients between both instruments ranged from 0.91 to 0.98 for the five classic bands.

    CONCLUSION: Both instruments demonstrated good analytical performance characterized by high precision, reproducibility and correlation.

    Matched MeSH terms: Blood Protein Electrophoresis/methods
  13. Fulltext Cell patterning for liver tissue engineering via dielectrophoretic mechanisms
    Yahya WN, Kadri NA, Ibrahim F
    Sensors (Basel), 2014 Jul 02;14(7):11714-34.
    PMID: 24991941 DOI: 10.3390/s140711714
    Liver transplantation is the most common treatment for patients with end-stage liver failure. However, liver transplantation is greatly limited by a shortage of donors. Liver tissue engineering may offer an alternative by providing an implantable engineered liver. Currently, diverse types of engineering approaches for in vitro liver cell culture are available, including scaffold-based methods, microfluidic platforms, and micropatterning techniques. Active cell patterning via dielectrophoretic (DEP) force showed some advantages over other methods, including high speed, ease of handling, high precision and being label-free. This article summarizes liver function and regenerative mechanisms for better understanding in developing engineered liver. We then review recent advances in liver tissue engineering techniques and focus on DEP-based cell patterning, including microelectrode design and patterning configuration.
    Matched MeSH terms: Electrophoresis/methods*
  14. Gelsolin and ceruloplasmin as potential predictive biomarkers for cervical cancer by 2D-DIGE proteomics analysis
    Lokamani I, Looi ML, Md Ali SA, Mohd Dali AZ, Ahmad Annuar MA, Jamal R
    Pathol Oncol Res, 2014 Jan;20(1):119-29.
    PMID: 23925487 DOI: 10.1007/s12253-013-9670-9
    This study aimed to identify candidate proteins which may serve as potential biological markers for cervical cancer using 2D-DIGE. Serum samples of controls, patients with cervical intraepithelial neoplasia grade 3 (CIN 3), squamous cell carcinoma of early (SCC I and II) and late (SCC III and IV) stage were subjected to 2D-DIGE. Differentially expressed spots were identified by tandem mass spectrometry. Validation of candidate proteins in serum and tissue samples were then performed by ELISA and immunohistochemistry (IHC) analysis respectively. A total of 20 differentially expressed proteins were identified. These proteins were found to play key roles in the apoptosis pathway, complement system, various types of transportation such as hormones, fatty acids, lipid, vitamin E and drug transportation, coagulation cascade, regulation of iron and immunologic response. Based on their functional relevancy to the progression of various cancers, 4 proteins namely the complement factor H, CD5-like antigen, gelsolin and ceruloplasmin were chosen for further validation using ELISA. Biological network analysis showed that ceruloplasmin and gelsolin are closely interacted with the oncogene NF-κb. These two proteins were further validated using the IHC. Gelsolin and ceruloplasmin may serve as potential predictive biomarkers for the progression of high grade lesions.
    Matched MeSH terms: Two-Dimensional Difference Gel Electrophoresis/methods
  15. Fulltext Identification of actinomycete communities in Antarctic soil from Barrientos Island using PCR-denaturing gradient gel electrophoresis
    Learn-Han L, Yoke-Kqueen C, Shiran MS, Vui-Ling CM, Nurul-Syakima AM, Son R, et al.
    Genet. Mol. Res., 2012;11(1):277-91.
    PMID: 22370930 DOI: 10.4238/2012.February.8.3
    The diversity of specific bacteria taxa, such as the actinomycetes, has not been reported from the Antarctic island of Barrientos. The diversity of actinomycetes was estimated with two different strategies that use PCR-denaturing gradient gel electrophoresis. First, a PCR was applied, using a group-specific primer that allows selective amplification of actinomycete sequences. Second, a nested-PCR approach was used that allows the estimation of the relative abundance of actinomycetes within the bacterial community. Molecular identification, which was based on 16S rDNA sequence analysis, revealed eight genera of actinomycetes, Actinobacterium, Actinomyces, an uncultured Actinomycete, Streptomyces, Leifsonia, Frankineae, Rhodococcus, and Mycobacterium. The uncultured Actinomyces sp and Rhodococcus sp appear to be the prominent genera of actinomycetes in Barrientos Island soil. PCR-denaturing gradient gel electrophoresis patterns were used to look for correlations between actinomycete abundance and environmental characteristics, such as type of rookery and vegetation. There was a significant positive correlation between type of rookery and abundance of actinomycetes; soil samples collected from active chinstrap penguin rookeries had the highest actinomycete abundance. Vegetation type, such as moss, which could provide a microhabitat for bacteria, did not correlate significantly with actinomycete abundance.
    Matched MeSH terms: Denaturing Gradient Gel Electrophoresis/methods
  16. Fabrication and mechanical properties of Al2O3/SiC/ZrO2 functionally graded material by electrophoretic deposition
    Askari E, Mehrali M, Metselaar IH, Kadri NA, Rahman MM
    J Mech Behav Biomed Mater, 2012 Aug;12:144-50.
    PMID: 22732480 DOI: 10.1016/j.jmbbm.2012.02.029
    This study describes the synthesis of Al(2)O(3)/SiC/ZrO(2) functionally graded material (FGM) in bio-implants (artificial joints) by electrophoretic deposition (EPD). A suitable suspension that was based on 2-butanone was applied for the EPD of Al(2)O(3)/SiC/ZrO(2), and a pressureless sintering process was applied as a presintering. Hot isostatic pressing (HIP) was used to densify the deposit, with beneficial mechanical properties after 2 h at 1800 °C in Ar atmosphere. The maximum hardness in the outer layer (90 vol.% Al(2)O(3)+10 vol.% SiC) and maximum fracture toughness in the core layer (75 vol.% Al(2)O(3)+10 vol.% SiC + 15 vol.% ZrO(2)) composite were 20.8±0.3 GPa and 8±0.1 MPa m(1/2), respectively. The results, when compared with results from Al(2)O(3)/ZrO(2) FGM, showed that SiC increased the compressive stresses in the outer layers, while the inner layers were under a residual tensile stress.
    Matched MeSH terms: Electrophoresis/methods*
  17. Isozymes of a rattan species and their genetic interpretation
    Bon MC
    Electrophoresis, 1996 Jul;17(7):1248-52.
    PMID: 8855412
    A combination of a modified Feret' (Silvae Genet. 1971, 20, 46-50) extraction buffer and two types of electrophoresis with acrylamide and starch gels were used to characterize allozymes in mature vegetative tissue of a commercially high value species of rattans (Calamus subinermis). From the analysis of allelic segregation from single maternal rattans and their offspring, genetic control of the 16 observed banding zones, which were consistently scorable, was assumed. Seventeen gene loci were identified. The percentage of polymorphic loci within Calamus subinermis was much higher (70.5%) than expected levels of genetic diversity for tropical woody and non-woody species. It is thought that the protocol described may be applied to the analysis of the genetic diversity of all the endangered Calamus species.
    Matched MeSH terms: Electrophoresis/methods*
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