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Prevalence and risk factors for positive lymphatic filariasis antibody in Sabah, Malaysia: a cross-sectional study

Zakaria ND, Avoi R

Trans R Soc Trop Med Hyg, 2021 Aug 28.
PMID: 34453834 DOI: 10.1093/trstmh/trab132

BACKGROUND: Lymphatic filariasis (LF) is a public health problem in Sabah, Malaysia. In the subdistrict of Tangkarason, nine rounds of mass drug administration (MDA) were probably not effective in reducing the prevalence of microfilaria to the <1% threshold recommended by the World Health Organization for stopping MDA. This cross-sectional study was conducted to identify the risk factors associated with positive LF antibody in Tangkarason.
METHODS: Eligible adults >18 y of age in seven endemic villages in Tangkarason, Beluran, Sabah, were interviewed and tested for LF antibody using the Brugia Rapid kit. Multivariable logistic regression was employed to analyse the associated factors.
RESULTS: A total of 244 respondents were included in this study. Their median age was 40 y (interquartile range 30-53). The antibody prevalence of brugian filariasis in the study population was 31.1% (95% confidence interval [CI] 25.7 to 37.2). Older age (adjusted odds ratio [aOR] 1.04 [95% CI 1.02 to 1.06]) and outdoor jobs (aOR 2.26 [95% CI 1.05 to 4.85]) were identified as independent risk factors for positive LF antibody. Participating in the MDA program previously (aOR 0.24 [95% CI 0.10 to 0.57]) was found to be a protective factor for LF infection.
CONCLUSIONS: A high prevalence of microfilariae was confirmed in all the study sites, which was above the target of <1%. Important factors associated with positive LF antibody were identified, which could be used as a guide for program managers to design more focused control measures in LF-endemic areas.
Fulltext Synthesis and evaluation of a molecularly imprinted polymer for 2,4-dinitrophenol

Zakaria ND, Yusof NA, Haron J, Abdullah AH

Int J Mol Sci, 2009 Jan;10(1):354-65.
PMID: 19333450 DOI: 10.3390/ijms10010354

Molecular imprinted polymers (MIP) are considered one of the most promising selective and novel separation methods for removal phenolic compound in wastewater treatment. MIP are crosslinked polymeric materials that exhibit high binding capacity and selectivity towards a target molecule (template), purposely present during the synthesis process. In this work MIP were prepared in a bulk polymerization method in acetonitrile using 2,4-dinitrophenol, acrylamide, ethylene glycol dimethacrylate, and benzoyl peroxide as template, functional monomer, cross-linker and initiator, respectively. An adsorption process for removal of nitrophenol using the fabricated MIP was evaluated under various pH and time conditions. The parameters studied for 2,4-dinitrophenol includes adsorption kinetics, adsorption isotherm, and selectivity. The maximum adsorption of nitrophenol by the fabricated MIP was 3.50 mg/g. The adsorption of 2,4-dinitrophenol by the fabricated MIP was found effective at pH 6.0. A kinetics study showed that nitrophenol adsorption follows a second order adsorption rate and the adsorption isotherm data is explained well by the Langmuir model.
Development of rapid gold nanoparticles based lateral flow assays for simultaneous detection of Shigella and Salmonella genera

Yahaya ML, Zakaria ND, Noordin R, Abdul Razak K

Biotechnol Appl Biochem, 2021 Oct;68(5):1095-1106.
PMID: 32935878 DOI: 10.1002/bab.2029

Salmonella and Shigella genera are common pathogens that contaminate foods and beverages. Lateral flow assays (LFA) are commonly used to detect these pathogens. However, most of the developed LFAs are for single detection. Simultaneous detection of pathogens is required to reduce cost and time. In this work, 40 nm gold nanoparticles (AuNPs) were synthesized using the seeding growth method as labeling agent. The AuNPs were characterized and conjugated with mouse anti-Gram negative endotoxin antibody. The nitrocellulose membrane HF135 was immobilized with anti-mouse IgG antibody as a control line and two separate test lines with either anti-Shigella or anti-Salmonella antibody, respectively. Color intensity of test lines was observed for positive samples. A milk sample was used as proof of concept to mimic actual contamination. The limit of detection of the LFA was 3.0 × 106 CFU/mL for multiplex detection of Shigella flexneri and Salmonella Typhi and for both single detections. The result was comparable with the enzyme-linked immunosorbent assay (ELISA) analysis. The produced LFA could differentiate between Shigella flexneri, Shigella boydii, Salmonella Enteritidis, and Salmonella Typhi. The developed LFA was able to identify Shigella flexneri and Salmonella Typhi with good sensitivity in milk samples, thus, beneficial to ensure the safety of food before entering the market.
Fulltext Simultaneous Sensing of Cd(II), Pb(II), and Cu(II) Using Gold Nanoparticle-Modified APTES-Functionalized Indium Tin Oxide Electrode: Effect of APTES Concentration

Mohamad Nor N, Nasrul SN, Zakaria ND, Abdul Razak K

ACS Omega, 2023 May 16;8(19):16587-16599.
PMID: 37214679 DOI: 10.1021/acsomega.2c07085

In this work, indium tin oxide (ITO) electrodes were functionalized with varying 3-aminopropyltriethoxysilane (APTES) concentration percentages (0.5, 0.75, 1.0, and 2.0 wt %) to obtain the optimum conditions for the assembly of the as-synthesized gold nanoparticles (AuNPs). The AuNP coverage, wettability, and electrochemical performance of the modified electrodes were evaluated. The AuNP/0.75% APTES-ITO-modified electrode exhibited uniform coverage of AuNPs and high electrochemical performance for the simultaneous detection of Cd(II), Pb(II), and Cu(II) ions. Under the optimum conditions, the AuNP/0.75% APTES-ITO-modified electrode showed a linear detection range of 5-120 ppb and limit of detection of 0.73, 0.90, and 0.49 ppb for the simultaneous detection of Cd(II), Pb(II), and Cu(II) ions, respectively, via square wave anodic stripping voltammetry. The modified electrode demonstrated good anti-interference toward other heavy metal ions, good reproducibility, and suitability for application in environmental sample analysis.
Fulltext Lateral flow test using Echinococcus granulosus native antigen B and comparison of IgG and IgG4 dipsticks for detection of human cystic echinococcosis

Khalilpour A, Sadjjadi SM, Moghadam ZK, Yunus MH, Zakaria ND, Osman S, et al.

Am J Trop Med Hyg, 2014 Nov;91(5):994-9.
PMID: 25200268 DOI: 10.4269/ajtmh.14-0170

Cystic echinococcosis (CE) caused by infection with Echinococcus granulosus is of major concern for humans in many parts of the world. Antigen B was prepared from E. granulosus hydatid fluid, and Western blots confirmed eight batches showing a band corresponding to the 8-/12-kDa subunit with positive serum and no low-molecular mass band (< 15 kDa) with negative serum. The batches were pooled and used to prepare lateral flow immunoglobulin G4 (IgG4) and IgG dipsticks. Diagnostic sensitivity was determined using serum samples from 21 hydatidosis patients, and diagnostic specificity was established using sera from 17 individuals infected with other parasites and 15 healthy people. IgG4 dipstick had a diagnostic sensitivity of 95% (20 of 21) and a specificity of 100% (32 of 32). The IgG dipstick had a sensitivity of 100% (21 of 21) and a specificity of 87.5% (28 of 32). Thus, both IgG and IgG4 dipsticks had high sensitivities, but IgG4 had greater specificity for the diagnosis of human CE.
Fulltext Production of recombinant Entamoeba histolytica pyruvate phosphate dikinase and its application in a lateral flow dipstick test for amoebic liver abscess

Saidin S, Yunus MH, Zakaria ND, Razak KA, Huat LB, Othman N, et al.

BMC Infect Dis, 2014 Apr 04;14:182.
PMID: 24708664 DOI: 10.1186/1471-2334-14-182

BACKGROUND: Amoebic liver abscess (ALA) is the most common clinical manifestation of extraintestinal amoebiasis especially in developing countries, causing up to 100 000 fatal cases annually. Accurate and early diagnosis is important to prevent the disease complications, however its diagnosis still poses many challenges due to the limitations of the available detection tools. Pyruvate phosphate dikinase (PPDK), an excretory-secretory protein of E. histolytica, has been reported as a potential diagnostic marker for ALA, hence it may be exploited in the development of a new test for ALA.
METHODS: Recombinant PPDK (rPPDK) was expressed, purified and evaluated by Western blot. In parallel, recombinant galactose-and-N-acetyl-D-galactosamine inhibitable lectin (Gal/GalNAc lectin) was produced and tested similarly. The protein identity was confirmed by analysis using MALDI-TOF/TOF. A lateral flow dipstick (LFD) test using rPPDK was subsequently developed (rPPDK-LFD) and evaluated for serodiagnosis of ALA.
RESULTS: rPPDK was expressed as soluble protein after 4 hours of induction with 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) at 30°C. Purification using nickel-nitrilotriacetic acid (Ni-NTA) resin yielded 1.5 mg of rPPDK from 1 L of culture with estimated molecular mass of 98 kDa on SDS-PAGE. Western blots using sera from patients with ALA, healthy individuals and other diseases probed with anti-human IgG4-HRP showed the highest sensitivity (93.3%) and specificity (100%); as compared to blots using IgG and IgG1 as secondary antibodies. Moreover, rPPDK showed better specificity when compared to rGal/GalNAc lectin. In the development of the LFD test, the optimum amount of rPPDK was 0.625 μg per dipstick and the optimum working concentration of colloidal gold conjugated anti-human IgG4 was optical density (OD) 5 (1.7 μg of anti-human IgG4). Evaluation of rPPDK-LFD using ALA patients and controls serum samples showed 87% diagnostic sensitivity and 100% specificity.
CONCLUSION: The developed rPPDK-LFD showed good potential for rapid diagnosis of ALA, and merit further multicentre validation using larger number of serum samples.
The effects of size and synthesis methods of gold nanoparticle-conjugated MαHIgG4 for use in an immunochromatographic strip test to detect brugian filariasis

Makhsin SR, Razak KA, Noordin R, Zakaria ND, Chun TS

Nanotechnology, 2012 Dec 14;23(49):495719.
PMID: 23164811 DOI: 10.1088/0957-4484/23/49/495719

This study describes the properties of colloidal gold nanoparticles (AuNPs) with sizes of 20, 30 and 40 nm, which were synthesized using citrate reduction or seeding-growth methods. Likewise, the conjugation of these AuNPs to mouse anti-human IgG(4) (MαHIgG(4)) was evaluated for an immunochromatographic (ICG) strip test to detect brugian filariasis. The morphology of the AuNPs was studied based on the degree of ellipticity (G) of the transmission electron microscopy images. The AuNPs produced using the seeding-growth method showed lower ellipticity (G ≤ 1.11) as compared with the AuNPs synthesized using the citrate reduction method (G ≤ 1.18). Zetasizer analysis showed that the AuNPs that were synthesized using the seeding-growth method were almost monodispersed with a lower polydispersity index (PDI; PDI≤0.079), as compared with the AuNPs synthesized using the citrate reduction method (PDI≤0.177). UV-visible spectroscopic analysis showed a red-shift of the absorbance spectra after the reaction with MαHIgG(4), which indicated that the AuNPs were successfully conjugated. The optimum concentration of the BmR1 recombinant antigen that was immobilized on the surface of the ICG strip on the test line was 1.0 mg ml(-1). When used with the ICG test strip assay and brugian filariasis serum samples, the conjugated AuNPs-MαHIgG(4) synthesized using the seeding-growth method had faster detection times, as compared with the AuNPs synthesized using the citrate reduction method. The 30 nm AuNPs-MαHIgG(4), with an optical density of 4 from the seeding-growth method, demonstrated the best performance for labelling ICG strips because it displayed the best sensitivity and the highest specificity when tested with serum samples from brugian filariasis patients and controls.
Fulltext Synthesis and characterization of molecularly imprinted polymer membrane for the removal of 2,4-dinitrophenol

Yusof NA, Zakaria ND, Maamor NA, Abdullah AH, Haron MJ

Int J Mol Sci, 2013;14(2):3993-4004.
PMID: 23429189 DOI: 10.3390/ijms14023993

Molecularly imprinted polymers (MIPs) were prepared by bulk polymerization in acetonitrile using 2,4-dinitrophenol, acrylamide, ethylene glycol dimethacrylate, and benzoyl peroxide, as the template, functional monomer, cross-linker, and initiator, respectively. The MIP membrane was prepared by hybridization of MIP particles with cellulose acetate (CA) and polystyrene (PS) after being ground and sieved. The prepared MIP membrane was characterized using Fourier transform infrared spectroscopy and scanning electron microscopy. The parameters studied for the removal of 2,4-dinitrophenol included the effect of pH, sorption kinetics, and the selectivity of the MIP membrane. Maximum sorption of 2,4-nitrophenol by the fabricated CA membrane with MIP (CA-MIP) and the PS membrane with MIP (PS-MIP) was observed at pH 7.0 and pH 5.0, respectively. The sorption of 2,4-dinitrophenol by CA-MIP and PS-MIP followed a pseudo-second-order kinetic model. For a selectivity study, 2,4-dichlorophenol, 3-chlorophenol, and phenol were selected as potential interferences. The sorption capability of CA-MIP and PS-MIP towards 2,4-dinitrophenol was observed to be higher than that of 2,4-dichlorophenol, 3-chlorophenol, or phenol.
Fulltext Effect of Supporting Background Electrolytes on the Nanostructure Morphologies and Electrochemical Behaviors of Electrodeposited Gold Nanoparticles on Glassy Carbon Electrode Surfaces

Zakaria ND, Omar MH, Ahmad Kamal NN, Abdul Razak K, Sönmez T, Balakrishnan V, et al.

ACS Omega, 2021 Sep 28;6(38):24419-24431.
PMID: 34604624 DOI: 10.1021/acsomega.1c02670

Electrodeposition is an electrochemical method employed to deposit stable and robust gold nanoparticles (AuNPs) on electrode surfaces for creating chemically modified electrodes (CMEs). The use of several electrodeposition techniques with different experimental parameters allow in obtaining various surface morphologies of AuNPs deposited on the electrode surface. By considering the electrodeposition of AuNPs in various background electrolytes could play an important strategy in finding the most suitable formation of the electrodeposited AuNP films on the electrode surface. This is because different electrode roughnesses can have different effects on the electrochemical activities of the modified electrodes. Thus, in this study, the electrodeposition of AuNPs onto the glassy carbon (GC) electrode surfaces in various aqueous neutral and acidic electrolytes was achieved by using the cyclic voltammetry (CV) technique with no adjustable CV parameters. Then, surface morphologies and electrochemical activities of the electrodeposited AuNPs were investigated using scanning electron microscopy (SEM), atomic force microscopy (AFM), CV, and electrochemical impedance spectroscopy (EIS). The obtained SEM and 3D-AFM images show that AuNPs deposited at the GC electrode prepared in NaNO3 solution form a significantly better, uniform, and homogeneous electrodeposited AuNP film on the GC electrode surface with nanoparticle sizes ranging from ∼36 to 60 nm. Meanwhile, from the electrochemical performances of the AuNP-modified GC electrodes, characterized by using a mixture of ferricyanide and ferrocyanide ions [Fe(CN6)3-/4-], there is no significant difference observed in the case of charge-transfer resistances (R ct) and heterogeneous electron-transfer rate constants (k o), although there are differences in the surface morphologies of the electrodeposited AuNP films. Remarkably, the R ct values of the AuNP-modified GC electrodes are lower than those of the bare GC electrode by 18-fold, as the R ct values were found to be ∼6 Ω (p < 0.001, n = 3). This has resulted in obtaining k o values of AuNP-modified GC electrodes between the magnitude of 10-2 and 10-3 cm s-1, giving a faster electron-transfer rate than that of the bare GC electrode (10-4 cm s-1). This study confirms that using an appropriate supporting background electrolyte plays a critical role in preparing electrodeposited AuNP films. This approach could lead to nanostructures with a more densely, uniformly, and homogeneously electrodeposited AuNP film on the electrode surfaces, albeit utilizing an easy and simple preparation method.
Fulltext Self-Assembled Iron Oxide Nanoparticle-Modified APTES-ITO Electrode for Simultaneous Stripping Analysis of Cd(II) and Pb(II) Ions

Mohamad Nor N, Arivalakan S, Zakaria ND, Nilamani N, Lockman Z, Abdul Razak K

ACS Omega, 2022 Feb 01;7(4):3823-3833.
PMID: 35128290 DOI: 10.1021/acsomega.1c07158

Carboxyl (-COOH)-stabilized iron oxide nanoparticles (IONPs) synthesized through co-precipitation were used to modify an indium tin oxide (ITO) electrode, which was chemically functionalized with 3-aminopropyltriethoxysilane (APTES) for heavy metal detection. The effect of soaking time (30, 60, 90, and 120 min) of IONP-COOH self-assembled on an APTES-ITO electrode was studied. Cyclic voltammetry and scanning electron microscopy were applied to analyze the electrochemical properties and morphologies of IONP-COOH/APTES-ITO modified electrode. The modified electrodes were then employed for the simultaneous detection of Cd(II) and Pb(II) by using square wave anodic stripping voltammetry. At 90 min of soaking time, excellent electrochemical performance and larger effective surface area (A e) were obtained. The linear range for the simultaneous detection of Cd(II) and Pb(II) ions using the modified electrode was 10-100 ppb with limits of detection of 0.90 and 0.60 ppb, respectively. The interference study revealed a low interference effect from Cr(III), Hg(II), Zn(II), Cu(II), Mg(II), Na(I), and K(I) toward the simultaneous detection of Cd(II) and Pb(II). Finally, the IONP-COOH/APTES-ITO-modified electrode was applied to analyze seawater samples and was able to simultaneously detect Cd(II) and Pb(II) ions.

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