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  1. Methylene blue removal by using pectin-based hydrogels extracted from dragon fruit peel waste using gamma and microwave radiation polymerization techniques
    Abdullah MF, Azfaralariff A, Lazim AM
    J Biomater Sci Polym Ed, 2018 10;29(14):1745-1763.
    PMID: 29989528 DOI: 10.1080/09205063.2018.1489023
    This research aims to compare the ability of smart hydrogel in removing the methylene blue prepared by using two different radiation methods. The extracted pectin from the dragon fruit peel (Hylocereus polyrhizus) was used with acrylic acid (AA) to produce a polymerized hydrogel through gamma and microwave radiation. The optimum hydrogel swelling capacity was obtained by varying the dose of radiation, pectin to AA ratio and pH used. From the array of samples, the ideal hydrogel was obtained at pH 8 with a ratio of 2:3 (pectin: AA) using 10 kGy and 400 W radiated gamma and microwave respectively. The performance of both hydrogels namely as Pc/AA(G) (gamma) and Pc/AA(Mw) (microwave) were investigated using methylene blue (MB) adsorption studies. In this study, three variables were manipulated, pH and MB concentration and hydrogel mass in order to find the optimum condition for the adsorption. Results showed that 20 mg of Pc/AA(G) performed the highest MB removal which was about 45% of 20 mg/L MB at pH 8. While 30 mg of Pc/AA(Mw) able to remove up to 35% of 20 mg/L MB at the same pH condition. To describe the adsorption mechanism, both kinetic models pseudo-first-order, pseudo-second-order were employed. The results from kinetic data showed that it fitted the pseudo-first-order as compared to pseudo-second-order model equation. This study provides alternative of green, facile and affective biomaterial for dye absorbents that readily available.
  2. Improving the efficiency of post-digestion method in extracting microplastics from gastrointestinal tract and gills of fish
    Jaafar N, Musa SM, Azfaralariff A, Mohamed M, Yusoff AH, Lazim AM
    Chemosphere, 2020 Dec;260:127649.
    PMID: 32688323 DOI: 10.1016/j.chemosphere.2020.127649
    Post-digestion treatment is an important step during sample preparation to facilitate the removal of undigested materials for better detection of ingested microplastics. Sieving, density separation with zinc chloride solution (ZnCl2), and oil extraction protocol (OEP) have been introduced in separating microplastics from sediments. The clean-up methods are rarely highlighted in previous studies, especially in the separation of microplastics from marine biota. Thus, this study proposed and compared the suitability of three techniques, which can reduce the number of undigested particles from the digestate of GIT and gills. Our result has shown excellent removal of non-plastics materials and reduces the coloration of filter paper in all treated samples. Both sieving and density separation achieved optimum post-digestion efficiencies of >95% for both GIT and gill samples, which former showed no effect on polymer integrity. Additionally, high recovery rate was obtained for the larger size microplastics (>500 μm) with approximately 97.7% (GIT) and 95.7% (gill), respectively. Exposure to the ZnCl2 solution led to a significant loss of smaller size PET and changed the absorption spectrums of all tested polymers. Particle morphology determined by SEM revealed such exposure eroded the surface of PET fragments and elemental analysis has shown detectable peaks of zinc and chlorine appeared. Low microplastics recoveries were achieved through OPE and residue of oil was observed from the infrared spectrum of all tested polymer. The findings demonstrate sieving with size fractioning can provide exceptional removal of non-plastics materials from the digestate of GIT and gill samples.
  3. Occurrence, distribution and characteristics of microplastics in gastrointestinal tract and gills of commercial marine fish from Malaysia
    Jaafar N, Azfaralariff A, Musa SM, Mohamed M, Yusoff AH, Lazim AM
    Sci Total Environ, 2021 Aug 05;799:149457.
    PMID: 34375867 DOI: 10.1016/j.scitotenv.2021.149457
    Microplastics are tiny plastic particles with size below 5 mm, prevalence in marine environments and the occurrence have been reported in commercial marine fish worldwide. Microplastics' abilities to absorb various marine contaminants raised considerable concern on their role as a vector to spread harmful pollutants to the alienated environment. This study focussed on the occurrence of microplastics in gastrointestinal tract (GIT) and gills of 158 fishes across 16 species from two locations in Malaysia coastal waters. Microplastics were detected approximately 86% in the GIT and 92% in the gills of examined fish. High incident of microplastics was detected in fishes from the area that is close to an urban area with average microplastics incident reaching up to 9.88 plastics items/individuals. Meanwhile, only 5.17 microplastics per individual were recorded in fishes from a less urbanised area. Isolated microplastics comprised 80.2% of fibres, 17.7% of fragments and the remaining was derived from filaments (3.1%). Infrared and Raman spectroscopy analysis of selected microplastics revealed the chemical composition of microplastics which comprised of polyethene (PE), polypropylene (PP), acrylonitrile butadiene styrene (ABS), polystyrene (PS) and polyethylene terephthalates (PET). FESEM images indicate, different surface characteristics of microplastics as a result of environmental exposure. Further, elemental analysis using EDX for green PE fragments showed the uneven distribution of chromium (Cr) and iron (Fe) on the surface, suggesting the adherence of heavy metals on the surface of microplastics. Overall findings indicate the widespread distribution of microplastics in commercial marine fishes from Malaysia waters and could potentially lead to human exposure through fish consumption.
  4. Sago starch nanocrystal-stabilized Pickering emulsions: Stability and rheological behavior
    Azfaralariff A, Farahfaiqah F, Joe LS, Fazry S, Mohamed M, Nazar MF, et al.
    Int J Biol Macromol, 2021 Jul 01;182:197-206.
    PMID: 33774073 DOI: 10.1016/j.ijbiomac.2021.03.132
    This study presents the isolation of SNC from sago starch and its performance as proficient particle emulsifier. It highlights the impact of SNC on the stability and rheological properties of oil-in-water (o/w) emulsions. The percentage yield of the SNC obtained was equivalent to 25 ± 0.1% (w/w) with particle diameters ranging from 25 to 100 nm. A series of Pickering emulsion at different ratios of oil (5%-35% v/v) and SNC (1%-4% w/v) was prepared for further investigations. The mean droplet diameter of emulsions obtained was ranged from 19.12 to 35.96 μm, confirming the effects of both SNC and oil content on the droplet's diameter distribution. Formulations with 4.0 wt% of SNC exhibited the maximum stability against coalescence. Results obtained have justified that the SNC can be used as an alternative solid emulsifier in producing stable emulsion with desired properties for various applications.
  5. Mutated Shiitake extracts inhibit melanin-producing neural crest-derived cells in zebrafish embryo
    Mahmood I, Azfaralariff A, Mohamad A, Airianah OB, Law D, Dyari HRE, et al.
    Comp Biochem Physiol C Toxicol Pharmacol, 2021 Jul;245:109033.
    PMID: 33737223 DOI: 10.1016/j.cbpc.2021.109033
    The ability of natural extracts to inhibit melanocyte activity is of great interest to researchers. This study evaluates and explores the ability of mutated Shiitake (A37) and wildtype Shiitake (WE) extract to inhibit this activity. Several properties such as total phenolic (TPC) and total flavonoid content (TFC), antioxidant activity, effect on cell and component profiling were conducted. While having no significant differences in total phenolic content, mutation resulted in A37 having a TFC content (1.04 ± 0.7 mg/100 ml) compared to WE (0.86 ± 0.9 mg/100 ml). Despite that, A37 extract has lower antioxidant activity (EC50, A37 = 549.6 ± 2.70 μg/ml) than WE (EC50 = 52.8 ± 1.19 μg/ml). Toxicity tests on zebrafish embryos show that both extracts, stop the embryogenesis process when the concentration used exceeds 900 μg/ml. Although both extracts showed pigmentation reduction in zebrafish embryos, A37 extract showed no effect on embryo heartbeat. Cell cycle studies revealed that WE significantly affect the cell cycle while A37 not. Further tests found that these extracts inhibit the phosphorylation of Glycogen synthase kinase 3 β (pGSK3β) in HS27 cell line, which may explain the activation of apoptosis in melanin-producing cells. It was found that from 19 known compounds, 14 compounds were present in both WE and A37 extracts. Interestingly, the presence of decitabine in A37 extract makes it very potential for use in the medical application such as treatment of melanoma, skin therapy and even cancer.
  6. Fulltext Comprehensive computational target fishing approach to identify Xanthorrhizol putative targets
    Shahid M, Azfaralariff A, Law D, Najm AA, Sanusi SA, Lim SJ, et al.
    Sci Rep, 2021 01 15;11(1):1594.
    PMID: 33452398 DOI: 10.1038/s41598-021-81026-9
    Xanthorrhizol (XNT), is a bioactive compound found in Curcuma xanthorrhiza Roxb. This study aimed to determine the potential targets of the XNT via computational target fishing method. This compound obeyed Lipinski's and Veber's rules where it has a molecular weight (MW) of 218.37 gmol-1, TPSA of 20.23, rotatable bonds (RBN) of 4, hydrogen acceptor and donor ability is 1 respectively. Besides, it also has half-life (HL) values 3.5 h, drug-likeness (DL) value of 0.07, oral bioavailability (OB) of 32.10, and blood-brain barrier permeability (BBB) value of 1.64 indicating its potential as therapeutic drug. Further, 20 potential targets were screened out through PharmMapper and DRAR-CPI servers. Co-expression results derived from GeneMANIA revealed that these targets made connection with a total of 40 genes and have 744 different links. Four genes which were RXRA, RBP4, HSD11B1 and AKR1C1 showed remarkable co-expression and predominantly involved in steroid metabolic process. Furthermore, among these 20 genes, 13 highly expressed genes associated with xenobiotics by cytochrome P450, chemical carcinogenesis and steroid metabolic pathways were identified through gene ontology (GO) and KEGG pathway analysis. In conclusion, XNT is targeting multiple proteins and pathways which may be exploited to shape a network that exerts systematic pharmacological effects.
  7. Formulation and In-Vitro Characterisation of Cross-Linked Amphiphilic Guar Gum Nanocarriers for Percutaneous Delivery of Arbutin
    Bostanudin MF, Salam A, Mahmood A, Arafat M, Kaharudin AN, Sahudin S, et al.
    J Pharm Sci, 2021 Dec;110(12):3907-3918.
    PMID: 34403653 DOI: 10.1016/j.xphs.2021.08.014
    Nano-colloidal systems formulated from amphiphilically-modified polysaccharides (degree of modification 16.6%) are focus of prominent study due to their potential to augment active penetration across the skin. Here we report the synthesis of amphiphilically-modified guar gum (GBE-GG) prepared by grafting with glycidol butyl ether (GBE), which were subsequently formed into nanocarriers and loaded with α-arbutin (22.3% loading). The monodispersed and close-to-spherical nanocarriers (size range 239-297 nm) formed via cross-linking were adequately stable mainly at low temperature (4 °C) under physiological pH condition. α-arbutin was released from GBE-GG NPs in a more sustained manner and the release profiles can be accurately represented by the 1st order kinetic model. In-vitro interactions on immortalised human keratinocytes (HaCaT) cells revealed an increase in biological membrane permeability as well as the absence of cellular toxicity at application pertinent concentrations. No substantial haemolytic activity appeared and flow cytometry analysis revealed effective cellular uptake, suggesting their potential as promising nanocarriers for percutaneous delivery that warrants further comprehensive research.
  8. In silico study of missense variants of FANCA, FANCC and FANCG genes reveals high risk deleterious alleles predisposing to Fanconi anemia pathogenesis
    Shahid M, Azfaralariff A, Zubair M, Abdulkareem Najm A, Khalili N, Law D, et al.
    Gene, 2022 Feb 20;812:146104.
    PMID: 34864095 DOI: 10.1016/j.gene.2021.146104
    Among the 22 Fanconi anemia (FA) reported genes, 90% of mutational spectra were found in three genes, namely FANCA (64%), FANCC (12%) and FANCG (8%). Therefore, this study aimed to identify the high-risk deleterious variants in three selected genes (FANCA, FANCC, and FANCG) through various computational approaches. The missense variant datasets retrieved from the UCSC genome browser were analyzed for their pathogenicity, stability, and phylogenetic conservancy. A total of 23 alterations, of which 16 in FANCA, 6 in FANCC and one variant in FANCG, were found to be highly deleterious. The native and mutant structures were generated, which demonstrated a profound impact on the respective proteins. Besides, their pathway analysis predicted many other pathways in addition to the Fanconi anemia pathway, homologous recombination, and mismatch repair pathways. Hence, this is the first comprehensive study that can be useful for understanding the genetic signatures in the development of FA.
  9. Fulltext Phytochemicals and Biological Activities of Garcinia atroviridis: A Critical Review
    Shahid M, Law D, Azfaralariff A, Mackeen MM, Chong TF, Fazry S
    Toxics, 2022 Oct 29;10(11).
    PMID: 36355947 DOI: 10.3390/toxics10110656
    Garcinia atriviridis Griff ex T. Anders (G. atroviridis) is one of the well-known species of the genus Garicinia that is native to Thailand, Myanmar, Peninsular Malaysia, and India. G. atroviridis is a perennial medium-sized tree that has a wide range of values, from food to medicinal use. Different parts of G. atroviridis are a great source of bioactive substances that have a positive impact on health. The extracts or bioactive constituents from G. atroviridis have demonstrated various therapeutic functions, including antioxidant, antimicrobial, anticancer, anti-inflammatory, antihyperlipidemic, and anti-diabetic. In this paper, we provide a critical review of G. atroviridis and its bioactive constituents in the prevention and treatment of different diseases, which will provide new insight to explore its putative domains of research.
  10. Fulltext Screening of high-risk deleterious missense variations in the CYP1B1 gene implicated in the pathogenesis of primary congenital glaucoma: A comprehensive in silico approach
    Shahid M, Azfaralariff A, Tufail M, Hussain Khan N, Abdulkareem Najm A, Firasat S, et al.
    PeerJ, 2022;10:e14132.
    PMID: 36518267 DOI: 10.7717/peerj.14132
    BACKGROUND: Primary congenital glaucoma (PCG) is the most common subtype of glaucoma caused by defects in the cytochrome P450 1B1 (CYP1B1) gene. It is developing among infants in more than 80% of cases who exhibit impairments in the anterior chamber angle and the trabecular meshwork. Thus, a comprehensive in silico approach was performed to evaluate the effect of high-risk deleterious missense variations in the CYP1B1 gene.

    MATERIAL AND METHODS: All the information for CYP1B1 missense variants was retrieved from the dbSNP database. Seven different tools, namely: SIFT, PolyPhen-2, PROVEAN, SNAP2, PANTHER, PhD-SNP, and Predict-SNP, were used for functional annotation, and two packages, which were I-Mutant 2.0 and MUpro, were used to predict the effect of the variants on protein stability. A phylogenetic conservation analysis using deleterious variants was performed by the ConSurf server. The 3D structures of the wild-type and mutants were generated using the I-TASSER tool, and a 50 ns molecular dynamic simulation (MDS) was executed using the GROMACS webserver to determine the stability of mutants compared to the native protein. Co-expression, protein-protein interaction (PPI), gene ontology (GO), and pathway analyses were additionally performed for the CYP1B1 in-depth study.

    RESULTS: All the retrieved data from the dbSNP database was subjected to functional, structural, and phylogenetic analysis. From the conducted analyses, a total of 19 high-risk variants (P52L, G61E, G90R, P118L, E173K, D291G, Y349D, G365W, G365R, R368H, R368C, D374N, N423Y, D430E, P442A, R444Q, F445L, R469W, and C470Y) were screened out that were considered to be deleterious to the CYP1B1 gene. The phylogenetic analysis revealed that the majority of the variants occurred in highly conserved regions. The MD simulation analysis exhibited that all mutants' average root mean square deviation (RMSD) values were higher compared to the wild-type protein, which could potentially cause CYP1B1 protein dysfunction, leading to the severity of the disease. Moreover, it has been discovered that CYP1A1, VCAN, HSD17B1, HSD17B2, and AKR1C3 are highly co-expressed and interact with CYP1B1. Besides, the CYP1B1 protein is primarily involved in the metabolism of xenobiotics, chemical carcinogenesis, the retinal metabolic process, and steroid hormone biosynthesis pathways, demonstrating its multifaceted and important roles.

    DISCUSSION: This is the first comprehensive study that adds essential information to the ongoing efforts to understand the crucial role of genetic signatures in the development of PCG and will be useful for more targeted gene-disease association studies.

  11. Mini review of microplastic pollutions and its impact on the environment and human health
    Azfaralariff A, Mat Lazim A, Amran NH, Mukhtar NH, Bakri ND, Azrihan NN, et al.
    Waste Manag Res, 2023 Jul;41(7):1219-1226.
    PMID: 36883418 DOI: 10.1177/0734242X231155395
    In recent years, the environmental pollution of microplastics (MPs) has increasingly drawn our attention. MPs are small fragments of plastics that are commonly dispersed in the environment. The accumulation of environmental MPs is due to population growth and urbanization, while natural disasters such as hurricanes, flooding and human activity may influence their distribution. The leaching of chemicals from MPs raises a significant safety problem and environmental approaches aimed at reducing the use and recycling of plastics, with the replacement by bioplastics and wastewater treatment developments are called for. This summary also helps in demonstrating the connection between terrestrial and freshwater MPs and wastewater treatment plants as the major contributors to environmental MPs by discharges of sludge and effluent. More research on the classification, detection, characterization and toxicity of MPs are essential to enable greater options and solutions. Control initiatives need to intensify the comprehensive study of MP waste control and management information programmes in the fields of institutional engagement, technological research and development, legislation and regulation. A comprehensive quantitative analysis approach for MPs should be created in the future, and more reliable traceability analysis methods should be built to examine further its environmental activity and existence, where this should be done to improve scientific research on MP pollution in terrestrial, freshwater and marine environments and hence, develop more scientific and rational control policies.
  12. Formation of Antihyperlipidemic Nano-Ezetimibe from Volatile Microemulsion Template for Enhanced Dissolution Profile
    Saleem MA, Yasir Siddique M, Nazar MF, Khan SU, Ahmad A, Khan R, et al.
    Langmuir, 2020 07 14;36(27):7908-7915.
    PMID: 32551692 DOI: 10.1021/acs.langmuir.0c01016
    Nanostructures play an important role in targeting sparingly water-soluble drugs to specific sites. Because of the structural flexibility and stability, the use of template microemulsions (μEs) can produce functional nanopharmaceuticals of different sizes, shapes, and chemical properties. In this article, we report a new volatile oil-in-water (o/w) μE formulation comprising ethyl acetate/ethanol/brij-35/water to obtain the highly water-dispersible nanoparticles of an antihyperlipidemic agent, ezetimibe (EZM-NPs), to enhance its dissolution profile. A pseudoternary phase diagram was delineated in a specified brij-35/ethanol ratio (1:1) to describe the transparent, optically isotropic domain of the as-formulated μE. The water-dilutable μE formulation, comprising an optimum composition of ethyl acetate (18.0%), ethanol (25.0%), brij-35 (25.0%), and water (32.0%), showed a good dissolvability of EZM around 4.8 wt % at pH 5.2. Electron micrographs showed a fine monomodal collection of EZM-loaded μE droplets (∼45 nm) that did not coalesce even after lyophilization, forming small spherical EZM-NPs (∼60 nm). However, the maturity of nanodrug droplets observed through dynamic light scattering suggests the affinity of EZM to the nonpolar microenvironment, which was further supported through peak-to-peak correlation of infrared analysis and fluorescence measurements. Moreover, the release profile of the as-obtained EZM-nanopowder increased significantly >98% in 30 min, which indicates that a reduced drug concentration will be needed for capsules or tablets in the future and can be simply incorporated into the multidosage formulation of EZM.
  13. Fulltext Anti-breast cancer synthetic peptides derived from the Anabas testudineus skin mucus fractions
    Najm AAK, Azfaralariff A, Dyari HRE, Othman BA, Shahid M, Khalili N, et al.
    Sci Rep, 2021 11 30;11(1):23182.
    PMID: 34848729 DOI: 10.1038/s41598-021-02007-6
    Previous study has shown the antimicrobial activities of mucus protein extracted from Anabas testudineus. In this study, we are interested in characterizing the anticancer activity of the A. testudineus antimicrobial peptides (AMPs). The mucus was extracted, fractioned, and subjected to antibacterial activity testing to confirm the fish's AMPs production. The cytotoxic activity of each fraction was also identified. Fraction 2 (F2), which shows toxicity against MCF7 and MDA-MB-231 were sent for peptide sequencing to identify the bioactive peptide. The two peptides were then synthetically produced and subjected to cytotoxic assay to prove their efficacy against cancer cell lines. The IC50 for AtMP1 against MCF7 and MDA-MB-231 were 8.25 ± 0.14 μg/ml and 9.35 ± 0.25 μg/ml respectively, while for AtMP2 it is 5.89 ± 0.14 μg/ml and 6.97 ± 0.24 μg/ml respectively. AtMP1 and AtMP2 treatment for 48 h induced breast cancer cell cycle arrest and apoptosis by upregulating the p53, which lead to upregulate pro-apoptotic BAX gene and downregulate the anti-apoptotic BCL-2 gene, consequently, trigger the activation of the caspase-3. This interaction was supported by docking analysis (QuickDBD, HPEPDOCK, and ZDOCK) and immunoprecipitation. This study provided new prospects in the development of highly effective and selective cancer therapeutics based on antimicrobial peptides.
  14. Marantodes pumilum: Systematic computational approach to identify their therapeutic potential and effectiveness
    Azfaralariff A, Farahfaiqah F, Shahid M, Sanusi SA, Law D, Mohd Isa AR, et al.
    J Ethnopharmacol, 2022 Jan 30;283:114751.
    PMID: 34662662 DOI: 10.1016/j.jep.2021.114751
    ETHNOPHARMACOLOGICAL RELEVANCE: Marantodes pumilum (MP) herbs, locally known as Kacip Fatimah, are widely used traditionally to improve women's health. The herb is frequently used for gynecological issues such as menstrual problems, facilitating and quickening delivery, post-partum medication, treats flatulence and dysentery, and. MP extracts are thought to aid in the firming and toning of abdominal muscles, tighten breasts and vaginal muscles, and anti-dysmenorrhea. It also was used for the treatment of gonorrhea and hemorrhoids. As MP product has been produced commercially recently, more in-depth studies should be conducted. The presence of numerous active compounds in MP might provide a synergistic effect and potentially offer other health benefits than those already identified and known.

    AIM OF THE STUDY: This study aimed to use a computational target fishing approach to predict the possible therapeutic effect of Marantodes pumilum and evaluated their effectivity.

    MATERIALS AND METHODS: This study involves a computational approach to identify the potential targets by using target fishing. Several databases were used: PubChem database to obtain the chemical structure of interested compounds; Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) server and the SWISSADME web tool to identify and select the compounds having drug-likeness properties; PharmMapper was used to identify top ten target protein of the selected compounds and Online Mendelian Inheritance in Man (OMIM) was used to predict human genetic problems; the gene id of top-10 proteins was obtained from UniProtKB to be analyzed by using GeneMANIA server to check the genes' function and their co-expression; Gene Pathway established by Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) of the selected targets were analyzed by using EnrichR server and confirmed by using DAVID (The Database for Annotation, Visualization and Integrated Discovery) version 6.8 and STRING database. All the interaction data was analyzed by Cytoscape version 3.7.2 software. The protein structure of most putative proteins was obtained from the RCSB protein data bank. Thedocking analysis was conducted using PyRx biological software v0.8 and illustrated by BIOVIA Discovery Studio Visualizer version 20.1.0. As a preliminary evaluation, a cell viability assay using Sulforhodamine B was conducted to evaluate the potential of the predicted therapeutic effect.

    RESULTS: It was found that four studied compounds are highly correlated with three proteins: EFGR, CDK2, and ESR1. These proteins are highly associated with cancer pathways, especially breast cancer and prostate cancer. Qualitatively, cell proliferation assay conducted shown that the extract has IC50 of 88.69 μg/ml against MCF-7 and 66.51 μg/ml against MDA-MB-231.

    CONCLUSIONS: Natural herbs are one of the most common forms of complementary and alternative medicine, and they play an important role in disease treatment. The results of this study show that in addition to being used traditionally to maintain women's health, the use of Marantodes pumilum indirectly has the potential to protect against the development of cancer cells, especially breast cancer. Therefore, further research is necessary to confirm the potential of this plant to be used in the development of anti-cancer drugs, especially for breast cancer.

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