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Fulltext Characterization of α-Glucosidase Inhibitors from Clinacanthus nutans Lindau Leaves by Gas Chromatography-Mass Spectrometry-Based Metabolomics and Molecular Docking Simulation

Murugesu S, Ibrahim Z, Ahmed QU, Nik Yusoff NI, Uzir BF, Perumal V, et al.

Molecules, 2018 Sep 19;23(9).
PMID: 30235889 DOI: 10.3390/molecules23092402

BACKGROUND: Clinacanthus nutans (C. nutans) is an Acanthaceae herbal shrub traditionally consumed to treat various diseases including diabetes in Malaysia. This study was designed to evaluate the α-glucosidase inhibitory activity of C. nutans leaves extracts, and to identify the metabolites responsible for the bioactivity.
METHODS: Crude extract obtained from the dried leaves using 80% methanolic solution was further partitioned using different polarity solvents. The resultant extracts were investigated for their α-glucosidase inhibitory potential followed by metabolites profiling using the gas chromatography tandem with mass spectrometry (GC-MS).
RESULTS: Multivariate data analysis was developed by correlating the bioactivity, and GC-MS data generated a suitable partial least square (PLS) model resulting in 11 bioactive compounds, namely, palmitic acid, phytol, hexadecanoic acid (methyl ester), 1-monopalmitin, stigmast-5-ene, pentadecanoic acid, heptadecanoic acid, 1-linolenoylglycerol, glycerol monostearate, alpha-tocospiro B, and stigmasterol. In-silico study via molecular docking was carried out using the crystal structure Saccharomyces cerevisiae isomaltase (PDB code: 3A4A). Interactions between the inhibitors and the protein were predicted involving residues, namely LYS156, THR310, PRO312, LEU313, GLU411, and ASN415 with hydrogen bond, while PHE314 and ARG315 with hydrophobic bonding.
CONCLUSION: The study provides informative data on the potential α-glucosidase inhibitors identified in C. nutans leaves, indicating the plant's therapeutic effect to manage hyperglycemia.
Fulltext Identification of α-glucosidase inhibitors from Clinacanthus nutans leaf extract using liquid chromatography-mass spectrometry-based metabolomics and protein-ligand interaction with molecular docking

Murugesu S, Ibrahim Z, Ahmed QU, Uzir BF, Nik Yusoff NI, Perumal V, et al.

J Pharm Anal, 2019 Apr;9(2):91-99.
PMID: 31011465 DOI: 10.1016/j.jpha.2018.11.001

The present study used in vitro and in silico techniques, as well as the metabolomics approach to characterise α-glucosidase inhibitors from different fractions of Clinacanthus nutans. C. nutans is a medicinal plant belonging to the Acanthaceae family, and is traditionally used to treat diabetes in Malaysia. n-Hexane, n-hexane: ethyl acetate (1:1, v/v), ethyl acetate, ethyl acetate: methanol (1:1, v/v), and methanol fractions were obtained via partitioning of the 80% methanolic crude extract. The in vitro α-glucosidase inhibitory activity was analyzed using all the fractions collected, followed by profiling of the metabolites using liquid chromatography combined with mass spectrometry. The partial least square (PLS) statistical model was developed using the SIMCA P+14.0 software and the following four inhibitors were obtained: (1) 4,6,8-Megastigmatrien-3-one; (2) N-Isobutyl-2-nonen-6,8-diynamide; (3) 1',2'-bis(acetyloxy)-3',4'-didehydro-2'-hydro-β, ψ-carotene; and (4) 22-acetate-3-hydroxy-21-(6-methyl-2,4-octadienoate)-olean-12-en-28-oic acid. The in silico study performed via molecular docking with the crystal structure of yeast isomaltase (PDB code: 3A4A) involved a hydrogen bond and some hydrophobic interactions between the inhibitors and protein. The residues that interacted include ASN259, HID295, LYS156, ARG335, and GLY209 with a hydrogen bond, while TRP15, TYR158, VAL232, HIE280, ALA292, PRO312, LEU313, VAL313, PHE314, ARG315, TYR316, VAL319, and TRP343 with other forms of bonding.
Highly sensitive and selective acute myocardial infarction detection using aptamer-tethered MoS2 nanoflower and screen-printed electrodes

Vasudevan M, Tai MJY, Perumal V, Gopinath SCB, Murthe SS, Ovinis M, et al.

Biotechnol Appl Biochem, 2021 Dec;68(6):1386-1395.
PMID: 33140493 DOI: 10.1002/bab.2060

Acute myocardial infarction (AMI) is one of the leading causes of death worldwide. Cardiac troponin I (cTn1) is a commonly used biomarker for the diagnosis of AMI. Although there are various detection methods for the rapid detection of cTn1 such as optical, electrochemical, and acoustic techniques, electrochemical aptasensing techniques are commonly used because of their ease of handling, portability, and compactness. In this study, an electrochemical cTn1 biosensor, MoS2 nanoflowers on screen-printed electrodes assisted by aptamer, was synthesized using hydrothermal technique. Field emission scanning electron microscopy revealed distinct 2D nanosheets and jagged flower-like 3D MoS2 nanoflower structure, with X-ray diffraction analysis revealing well-stacked MoS2 layers. Voltammetry aptasensing of cTn1 ranges from 10 fM to 1 nM, with a detection limit at 10 fM and a sensitivity of 0.10 nA µM-1 cm-2 . This is a ∼fivefold improvement in selectivity compared with the other proteins and human serum. This novel aptasensor retained 90% of its biosensing activity after 6 weeks with a 4.3% RSD and is a promising high-performance biosensor for detecting cTn1.
Gold-Nanohybrid Biosensors for Analyzing Blood Circulating Clinical Biomacromolecules: Current Trend toward Future Remote Digital Monitoring

Letchumanan I, Gopinath SCB, Md Arshad MK, Mohamed Saheed MS, Perumal V, Voon CH, et al.

Crit Rev Anal Chem, 2020 Sep 08.
PMID: 32897761 DOI: 10.1080/10408347.2020.1812373

Mortality level is worsening the situation worldwide thru blood diseases and greatly jeopardizes the human health with poor diagnostics. Due to the lack of successful generation of early diagnosis, the survival rate is currently lower. To overcome the present hurdle, new diagnostic methods have been choreographed for blood disease biomarkers analyses with the conjunction of ultra-small ideal gold nanohybrids. Gold-hybrids hold varieties of unique features, such as high biocompatibility, increased surface-to-volume ratio, less-toxicity, ease in electron transfer and have a greater localized surface plasmon resonance. Gold-nanocomposites can be physically hybrid on the sensor surface and functionalize with the biomolecules using appropriate chemical conjugations. Revolutionizing biosensor platform can be prominently linked for the nanocomposite applications in the current research on medical diagnosis. This review encloses the new developments in diagnosing blood biomarkers by utilizing the gold-nanohybrids. Further, the current state-of-the-art and the future envision with digital monitoring for facile telediagnosis were narrated.
Fulltext In vitro antioxidant and, α-glucosidase inhibitory activities and comprehensive metabolite profiling of methanol extract and its fractions from Clinacanthus nutans

Alam MA, Zaidul IS, Ghafoor K, Sahena F, Hakim MA, Rafii MY, et al.

BMC Complement Altern Med, 2017 Mar 31;17(1):181.
PMID: 28359331 DOI: 10.1186/s12906-017-1684-5

BACKGROUND: This study was aimed to evaluate antioxidant and α-glucosidase inhibitory activity, with a subsequent analysis of total phenolic and total flavonoid content of methanol extract and its derived fractions from Clinacanthus nutans accompanied by comprehensive phytochemical profiling.
METHODS: Liquid-liquid partition chromatography was used to separate methanolic extract to get hexane, ethyl acetate, butanol and residual aqueous fractions. The total antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazy (DPPH) radical scavenging and ferric reducing antioxidant power assay (FRAP). The antidiabetic activity of methanol extract and its consequent fractions were examined by α-glucosidase inhibitory bioassay. The chemical profiling was carried out by gas chromatography coupled with quadrupole time-of-flight mass spectrometry (GC Q-TOF MS).
RESULTS: The total yield for methanol extraction was (12.63 ± 0.98) % (w/w) and highest fractionated value found for residual aqueous (52.25 ± 1.01) % (w/w) as compared to the other fractions. Significant DPPH free radical scavenging activity was found for methanolic extract (63.07 ± 0.11) % and (79.98 ± 0.31) % for ethyl acetate fraction among all the fractions evaluated. Methanol extract was the most prominent in case of FRAP (141.89 ± 0.87 μg AAE/g) whereas most effective reducing power observed in ethyl acetate fraction (133.6 ± 0.2987 μg AAE/g). The results also indicated a substantial α-glucosidase inhibitory activity for butanol fraction (72.16 ± 1.0) % and ethyl acetate fraction (70.76 ± 0.49) %. The statistical analysis revealed that total phenolic and total flavonoid content of the samples had the significant (p
Fulltext Laser-scribed graphene nanofiber decorated with oil palm lignin capped silver nanoparticles: a green biosensor

Tai MJY, Perumal V, Gopinath SCB, Raja PB, Ibrahim MNM, Jantan IN, et al.

Sci Rep, 2021 Mar 09;11(1):5475.
PMID: 33750861 DOI: 10.1038/s41598-021-85039-2

Tuberculosis (TB), caused by Mycobacterium tuberculosis (M. tuberculosis), requires a high level of attention and is one of the most infectious diseases in the air. Present methods of diagnosing TB remain ineffective owing to their low sensitivity and time consumption. In this study, we produced a green graphene nanofiber laser biosensor (LSG-NF) decorated with oil palm lignin-based synthetic silver nanoparticles (AgNPs). The resulting composite morphology was observed by field-emission scanning electron microscopy and transmission electron microscopy, which revealed the effective adaptation of the AgNPs to the LSG-NF surface. The successful attachment of AgNPs and LSG-NFs was also evident from X-ray diffraction and Raman spectroscopy studies. In order to verify the sensing efficiency, a selective DNA sample captured on AgNPs was investigated for specific binding with M.tb target DNA through selective hybridisation and mismatch analysis. Electrochemical impedance studies further confirmed sensitive detection of up to 1 fM, where a detection limit of 10-15 M was obtained by estimating the signal-to-noise ratio (S/N = 3:1) as 3σ. Successful DNA immobilisation and hybridisation was confirmed by the detection of phosphorus and nitrogen peaks based on X-ray photoelectron spectroscopy and Fourier-transform infrared spectroscopy. The stability and repeatability of the analysis were high. This approach provides an affordable potential sensing system for the determination of M. tuberculosis biomarker and thus provides a new direction in medical diagnosis.
Fulltext Characterization of Gold-Sputtered Zinc Oxide Nanorods-a Potential Hybrid Material

Perumal V, Hashim U, Gopinath SC, Rajintra Prasad H, Wei-Wen L, Balakrishnan SR, et al.

Nanoscale Res Lett, 2016 Dec;11(1):31.
PMID: 26787050 DOI: 10.1186/s11671-016-1245-8

Generation of hybrid nanostructures has been attested as a promising approach to develop high-performance sensing substrates. Herein, hybrid zinc oxide (ZnO) nanorod dopants with different gold (Au) thicknesses were grown on silicon wafer and studied for their impact on physical, optical and electrical characteristics. Structural patterns displayed that ZnO crystal lattice is in preferred c-axis orientation and proved the higher purities. Observations under field emission scanning electron microscopy revealed the coverage of ZnO nanorods by Au-spots having diameters in the average ranges of 5-10 nm, as determined under transmission electron microscopy. Impedance spectroscopic analysis of Au-sputtered ZnO nanorods was carried out in the frequency range of 1 to 100 MHz with applied AC amplitude of 1 V RMS. The obtained results showed significant changes in the electrical properties (conductance and dielectric constant) with nanostructures. A clear demonstration with 30-nm thickness of Au-sputtering was apparent to be ideal for downstream applications, due to the lowest variation in resistance value of grain boundary, which has dynamic and superior characteristics.
Fulltext Electrochemical biosensor detection on respiratory and flaviviruses

Ang PC, Perumal V, Ibrahim MNM, Adnan R, Mohd Azman DK, Gopinath SCB, et al.

Appl Microbiol Biotechnol, 2023 Mar;107(5-6):1503-1513.
PMID: 36719432 DOI: 10.1007/s00253-023-12400-y

Viruses have spread throughout the world and cause acute illness or death among millions of people. There is a growing concern about methods to control and combat early-stage viral infections to prevent the significant public health problem. However, conventional detection methods like polymerase chain reaction (PCR) requires sample purification and are time-consuming for further clinical diagnosis. Hence, establishing a portable device for rapid detection with enhanced sensitivity and selectivity for the specific virus to prevent further spread becomes an urgent need. Many research groups are focusing on the potential of the electrochemical sensor to become a key for developing point-of-care (POC) technologies for clinical analysis because it can solve most of the limitations of conventional diagnostic methods. Herein, this review discusses the current development of electrochemical sensors for the detection of respiratory virus infections and flaviviruses over the past 10 years. Trends in future perspectives in rapid clinical detection sensors on viruses are also discussed. KEY POINTS: • Respiratory related viruses and Flavivirus are being concerned for past decades. • Important to differentiate the cross-reactivity between the virus in same family. • Electrochemical biosensor as a suitable device to detect viruses with high performance.
Toxicity study on Clinacanthus nutans leaf hexane fraction using Danio rerio embryos

Murugesu S, Khatib A, Ahmed QU, Ibrahim Z, Uzir BF, Benchoula K, et al.

Toxicol Rep, 2019;6:1148-1154.
PMID: 31993329 DOI: 10.1016/j.toxrep.2019.10.020

Clinacanthus nutans, an herbal shrub belonging to the Acanthaceae family, is traditionally used as a functional food to treat various ailments in Malaysia and Indonesia. Although the polar fraction of this plant shows non-toxic effect, the toxicity of the non-polar extract is not reported so far. The present study aimed to assess the toxic effect and determine the lethal concentration of this non-polar fraction using zebrafish embryos. The n-hexane fraction was partitioned from the crude extract of C. nutans obtained using 80% methanolic solution. After spawning of the adult male and female zebrafish, the eggs were collected, transferred into a 96-well plate and incubated with the n-hexane fraction at concentrations of 15.63 μg/ml, 31.25 μg/ml, 62.5 μg/ml, 125 μg/ml, 250 μg/ml and 500 μg/ml in 2% DMSO. The survival and sublethal endpoint were assessed, the mortality and hatchability rates were calculated based on microscopic observation, while the heartbeat rate was measured using DanioScope software. The median lethal concentration (LC50) of the C. nutans n-hexane fraction, which was determined using probit analysis, was calculated to be 75.49 μg/mL, which is harmful. Moreover, gas chromatography-mass spectrometry (GC-MS) analysis revealed the presence of palmitic acid, phytol, hexadecanoic acid, 1-monopalmitin, stigmast-5-ene, pentadecanoic acid, heptadecanoic acid, 1-linolenoylglycerol and stigmasterol in the n-hexane fraction.
A quadruplet 3-D laser scribed graphene/MoS2, functionalised N2-doped graphene quantum dots and lignin-based Ag-nanoparticles for biosensing

Vasudevan M, Perumal V, Raja PB, Ibrahim MNM, Lee HL, Gopinath SCB, et al.

Int J Biol Macromol, 2023 Dec 31;253(Pt 2):126620.
PMID: 37683754 DOI: 10.1016/j.ijbiomac.2023.126620

Troponin I is a protein released into the human blood circulation and a commonly used biomarker due to its sensitivity and specificity in diagnosing myocardial injury. When heart injury occurs, elevated troponin Troponin I levels are released into the bloodstream. The biomarker is a strong and reliable indicator of myocardial injury in a person, with immediate treatment required. For electrochemical sensing of Troponin I, a quadruplet 3D laser-scribed graphene/molybdenum disulphide functionalised N2-doped graphene quantum dots hybrid with lignin-based Ag-nanoparticles (3D LSG/MoS2/N-GQDs/L-Ag NPs) was fabricated using a hydrothermal process as an enhanced quadruplet substrate. Hybrid MoS2 nanoflower (H3 NF) and nanosphere (H3 NS) were formed independently by varying MoS2 precursors and were grown on 3D LSG uniformly without severe stacking and restacking issues, and characterized by morphological, physical, and structural analyses with the N-GQDs and Ag NPs evenly distributed on 3D LSG/MoS2 surface by covalent bonding. The selective capture of and specific interaction with Troponin I by the biotinylated aptamer probe on the bio-electrode, resulted in an increment in the charge transfer resistance. The limit of detection, based on impedance spectroscopy, is 100 aM for both H3 NF and H3 NS hybrids, with the H3 NF hybrid biosensor having better analytical performance in terms of linearity, selectivity, repeatability, and stability.
Microwave welding with SiCNW/PMMA nanocomposite thin films: enhanced joint strength and performance

Foong PY, Voon CH, Lim BY, Teh PL, Yeoh CK, Parmin NA, et al.

Nanotechnology, 2025 Jan 20;36(11).
PMID: 39787627 DOI: 10.1088/1361-6528/ada7ff

Most previously reported susceptors for microwave welding are in powder form. In this study, a thin-film susceptor was employed due to its uniform heating rate and ease of handling. Silicon carbide nanowhisker (SiCNW) were incorporated into a poly(methyl methacrylate) (PMMA) matrix to create a nanocomposite thin film, which served as the susceptor. The microwave welding process involved three straightforward steps: fabrication of the PMMA/SiCNW nanocomposite thin film, application of the nanocomposite film to the target area, and subsequent microwave heating. Upon cooling, a robust microwave-welded joint was formed. The mechanical properties and microstructure of the welded joints were characterized using single-lap shear tests, three-point bending tests, and scanning electron microscopy. Results demonstrated that the shear strength and elastic modulus of the welded joints were optimized with increased heating time and SiCNW filler loading. This optimization is attributed to the formation of a SiCNW-filled polypropylene (PP) nanocomposite layer of increasing thickness at the welded joint interface. However, the incorporation of SiCNW also constrained the mobility of the PP chains, reducing the joint's flexibility. Furthermore, the welded joint formed with the PMMA/SiCNW nanocomposite thin-film susceptor exhibited an 18.82% improvement in shear strength compared to joints formed with a powdered SiCNW susceptor. This study not only demonstrates the potential of PMMA/SiCNW nanocomposite thin films as efficient susceptors for microwave welding but also paves the way for developing high-performance polymer-based composite joints with improved mechanical properties for applications in the automotive, aerospace, and construction industries.

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