Malaysian Journal of Biochemistry
& Molecular Biology
(E-ISSN: 2600-9005)
The Official Publication of the Malaysian Society for Biochemistry & Molecular Biology (MSBMB)
Indexed by SCOPUS and Malaysian Citation Index (MYCITE)
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December 2023
Malay. J. Biochem. Mol. Biol. (2023) 26 (2)
TABLE OF CONTENTS
Page 1- 5
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Aisyah Mohd Ismail, Wan Rozianoor Mohd Hassan, Abd Razzif Abd Razak and Farida Zuraina Mohd Yusof
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GENETIC TYPING OF THREE MALAYSIAN DURIAN VARIETIES USING SIMPLE SEQUENCE REPEAT (SSR) MARKERS
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Abstract
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There have been few studies on the molecular aspects of durian up to this point. Although morphological classification is practical, simple, and quick, it suffers from phenotypic plasticity due to environmental influences and age. Hence, the genetic characterization of durian varieties needs to be carried out. This study was performed to evaluate the effectiveness of three simple sequence repeat (SSR) markers to identify and discriminate three durian types: MK (Musang King), D24 and D101. Successful amplification of SSR regions were observed in the durian DNA samples. A total of 8 alleles were generated by all primers. Both DZ01 and DZ04 primers showed the highest number of polymorphic fragments. Among the three SSR primers, DZ03 is the least informative with only 2 number of alleles produced. The most important finding of this study was that each primer is unique and specific to one type of durian sample. This preliminary study showed that the SSR loci could be used as genetic markers to assist future durian breeding program.
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Page 6 - 30
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Hui Shi Saw, Bernard Kok Bang Lee, Hwei-San Loh
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GENE EXPRESSION MINING AND in silico ANALYSIS OF DIFFERENTIALLY EXPRESSED GENES IN HEAD AND NECK CANCER
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Abstract
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The advancement of high-throughput transcriptome profiling techniques, such as next-generation sequencing and microarray, has led to the development of bioinformatics tools and databases for functional genomics. Integrated bioinformatics analysis has emerged as a promising strategy to address the major cause of morbidity and death globally: cancer. In this study, we aimed to use an integrated bioinformatics pipeline to identify potential molecular biomarkers for diagnosis and prognosis in cancer studies. Specifically, we focused on head and neck squamous cell carcinoma (HNSCC). To achieve this, we performed a meta-analysis on expression datasets from the Gene Expression Omnibus (GEO) using GEO2R to derive differentially expressed genes (DEGs). Subsequently, we conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Protein-protein interaction networks of the up-regulated and down-regulated genes were constructed using the STRING database, and the top ten hub genes for each group were identified using cytoHubba. The relative mRNA expression of the identified DEGs was validated with GEPIA2, and their correlation with the overall survival of HNSCC patients was assessed using Kaplan-Meier analysis. Combining our findings with published evidence, we observed that the up-regulated genes primarily function in the extracellular matrix and cell cycle regulation. In contrast, the down-regulated genes are involved in muscle contraction. Our results suggest that six down-regulated genes (MYL1, MYL3, MYH6, MYLPF, ACTA1, TTN) and five up-regulated genes (CDC20, CCNB1, MAD2L1, TOP2A, MMP9) have the potential to serve as diagnosis biomarkers. In contrast, five up-regulated genes (FN1, CDK1, PLK1, AURKA, CD44) could be used for prognosis and diagnosis in the clinical analysis of HNSCC. This study demonstrated the effectiveness of an integrated bioinformatics approach in identifying clinically relevant biomarkers for HNSCC, and the pipeline could be applied to other cancer datasets. Further investigation of the identified biomarkers will enrich our understanding of their involvement in the molecular mechanism of carcinogenesis and provide potential therapeutic targets for HNSCC.
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Page 31 - 38
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Yusnaini Md Yusoff, Ahmad Firdhaus Arham, Noor Sharizad Rusly, Muhammad Firdaus Aziz, Mohd Rosly Shaari, Jasni Sabri, Shanmugavelu Sithambaram, Noordin Mohd Mustapha, Tan Sheau Wei, Nursyuhada Haron, Nurul Huda Mohd Zairi, Mohd Hakimi Mohd Kassim and Hazilawati Hamzah
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MODULATING THE BCL2 TO BAX RATIO: TURMERIC’S POTENTIAL IN LEUKAEMIA THERAPY
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Abstract
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Leukaemia ranks among the ten most prevalent types of cancer in Malaysia, exhibiting a death rate of 4.4%. Therefore, the objective of this study was to assess the preventative effects of a high dose of dried Curcuma longa (C. longa) rhizomes supplementation in rats with N-Methyl-N-Nitrosourea (MNU)-induced leukaemia. Two weeks were spent acclimatizing 64 mature male Sprague Dawley rats. At week 0, all rats were separated into A, B, C, and D groups. MNU was intraperitoneally given to Group C and D rats at 240 mg/kg. C. longa rhizomes were dried and given to Groups B and D rats at 5000 mg/kg. Group A rats were controls. The rats were killed in week 20. Blood samples were examined for the presence of leukaemic cells and underwent RNA extraction for quantitative real-time polymerase chain reaction (RT-PCR). By blast cell appearance, all rats in Group C had 100% leukaemia in the blood smear, while Group D had 88%. The ratio of Bcl-2 to Bax transcripts in blood was 3.3-fold (10.50±1.26) higher in Group C compared to the control rats (3.17±1.07), indicating high levels of anti-apoptotic cells caused by leukaemia. Interestingly, the ratio of Bcl-2 to Bax transcripts in Group D rats (3.58±0.82) was similar to the control rats. A quantitative RT-PCR experiment found that adding dried C. longa rhizome to a meal significantly reduced Bcl2 to Bax ratio in leukaemia rats, which significantly reduced the incidence of leukaemia.
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Page 39 - 44
Hao Ing Yeoh, Md Salzihan Md Salleh, Maya Mazuwin Yahya, Andee Dzulkarnaen Zakaria, Wan Mohd Nazri Wan Zainon, Ahmad Aizat Abdul Aziz
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GENETIC ASSOCIATION OF ABCB1 1236 C>T POLYMORPHISM ON MALAY TRIPLE NEGATIVE BREAST CANCER (TNBC) SUSCEPTIBILITY RISK
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Abstract
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Triple-negative breast cancer (TNBC) is characterized by the absence of estrogen receptors (ER), progesterone receptors (PR), and the lack of amplification of the human epidermal growth factor 2 (HER2) receptor. Adenosine triphosphate-binding cassette (ABC) subfamily B member 1 (ABCB1) serves as a drug efflux transporter, facilitating the translocation of various substrates (xenobiotics, toxins, carcinogens, etc.) across the membrane in an ATP-dependent manner. Genetic variations on ABCB1 may lead to reduced substrate specificity, stability, and gene expression, thereby influencing the efflux activity of the protein. These variations can ultimately impact an individual’s susceptibility to cancer. The present study aims to investigate the association of polymorphisms in ABCB1 (1236 C > T, 2677 G > T/A, and 3435 C > T) in modulating the individual susceptibility risk to TNBC. DNA was extracted from blood samples collected from 75 TNBC patients and 100 healthy controls. Genotyping was performed using the PCR-RFLP technique, and the resulting genotype patterns were categorized into homozygous wild type, heterozygous, and homozygous variants. The association between genotype and TNBC and clinicopathological variables was assessed using the independent χ2 test. The strength of the association was determined by calculating the odds ratio (OR) with a 95% confidence interval. Subsequently, linkage disequilibrium and haplotype association analyses were performed to evaluate the association of the ABCB1 haplotype with TNBC susceptibility. Overall, carriers of TT genotype and T allele of ABCB1 1236 C > T exhibited an increased OR of 2.750 (95% CI: 1.054–7.175) and 1.545 (95% CI: 1.001–2.385) for developing TNBC. Specifically, the ABCB1 1236 C > T variant is significantly associated with early age at diagnosis, advanced TNM staging, and the metaplastic/medullary subtype of carcinoma. Lastly, the haplotype 1236C/3435T/2677G was also associated with a reduced risk of TNBC. In summary, ABCB1 1236 C > T polymorphism was associated with an increased risk of TNBC susceptibility and correlated with early age diagnosis, high tumor staging, and the metaplastic/medullary subtype in TNBC patients. This suggests a potential predictive role in TNBC susceptibility and development for this polymorphism.
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Page 45 - 54
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Nik Ida Mardiana Nik-Pa, Mohamad Farhan Mohamad Sobri, Nurhasliza Zolkefli, Suraini Abd-Aziz, Mohamad Faizal Ibrahim, Noorjahan Banu Mohamed Alitheen, Norhayati Ramli
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DECIPHERING THE MOLECULAR LANDSCAPE: IN-SILICO ANALYSIS OF CYCLODEXTRIN GLYCOSYLTRANSFERASE FOR ENHANCED ENZYME FUNCTIONALITY AND CYCLODEXTRIN SYNTHESIS
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Abstract
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Cyclodextrin glycosyltransferase (CGTase) has always played a significant role in the production of cyclodextrin through the cyclization reaction. The wide application of this valuable protein demands a better understanding, leading to a comprehensive in-silico analysis of CGTase. The analysis focused on the functional domain composition of the recombinant CGT-BS protein by comparing it with several other CGTase proteins from different Bacillus spp. A three-dimensional (3-D) model was constructed to predict the active, substrate binding and cyclization sites of the CGT-BS protein. Structural function prediction revealed the active site within domain A at the wide end of the (β/α)8-barrel, with Asp 268, Glu 296 and Asp 357 as the catalytic residues. Additionally, the reaction site for cyclization was identified in domain B at Tyr 234. In comparison to maltose binding sites (MBS) 1 and 2 which are associated with raw starch binding activity, a comparable role is deduced for the MBS identified on the surface of the Domain E protein. We additionally observed that the residues Tyr 139, Arg 266 and Asp 367, located at the substrate binding cleft of the catalytic site, exhibited heightened hydrophobicity and concurrent cyclization activity. The successful extracellular expression of the CGT-BS protein is also anticipated to be facilitated by the presence of a functional signal peptide. In conclusion, our in-depth in-silico analysis unveils critical insights into the structural and functional aspects of CGT-BS protein, laying the groundwork for further exploration of its catalytic mechanisms and potential applications in cyclodextrin synthesis.
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Page 55 - 65
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Gayathiri Verasoundarapandian, Chiew Yen Wong, Zheng Syuen Lim, Khadijah Nabilah Mohd Zahri, Syazani Darham, Nur Nadhirah Zakaria and Siti Aqlima Ahmad
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OPTIMISATION OF ANTARCTIC FILAMENTOUS ALGA GROWTH IN THE PRESENCE OF MOLYBDENUM
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Abstract
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Elevated concentrations of heavy metals have been identified in Antarctica due to growing anthropogenic activities in recent years. Molybdenum (Mo) is a trace element that has not been extensively studied in terms of its toxicity towards the environment, especially in extremely cold weather. The algae communities in the Antarctic were less focused and explored, unlike indigenous bacteria consortia in their response to heavy metals. The study aims to optimise the physicochemical conditions for optimal growth of an Antarctic algal, Klebsormidium sp. in the presence of Mo via conventional one‒factor‒at‒a‒time (OFAT) and growth kinetics analysis. Algal cultures with aeration showed a higher growth rate (μ = 0.2352 d-1) than those without aeration (μ = 0.1976 d-1). Based on the optimised parameter, the overall biomass yields with and without aeration systems correspond to each other (P > 0.05). It was discovered that the Klebsormidium sp. showed maximal growth in terms of biomass at 20 g/L of sucrose, 2 g/L of ammonium nitrate, 4 g/L NaCl concentration and pH 7.5. The overall optimised conditions were further analysed using the Exponential growth model, which demonstrated no significant difference (P > 0.05) in the algae growth rate with aeration (0.020 ± 0.0018 h-1) and without aeration (0.020 ± 0.0015 h-1). The Antarctic filamentous algae exhibited the ability to grow in heavy metal, Mo at optimal growth conditions, but the aeration systems did not affect the algae growth significantly. Therefore, this study could help in understanding the capability of algae to grow in the presence of heavy metal through various manipulations of growth parameters and act as a preliminary study for bioremediation of Mo in Antarctic polluted sites.
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Page 66 - 75
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Jye Ping Fam, Suriana Sabri, Syarul Nataqain Baharum, Lorrine Eseoghene Okojie, Siti Nur Hazwani Oslan,
Abu Bakar Salleh, Siti Nurbaya Oslan
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FATTY ACID AS THE POTENTIAL INDUCER FOR RECOMBINANT LIPASE EXPRESSION IN Meyerozyma guilliermondii STRAIN SO
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Abstract
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High amount of methanol concentration in the cultivation medium during the production of enzymes in methylotrophic yeast could inhibit the cell growth, level of enzyme expressed as well as limit the use of the enzyme for certain foods and pharmaceutical production. A recombinant T1 lipase was expressed using Meyerozyma guilliermondii strain SO as a host under the regulation of alcohol oxidase promoter without methanol induction. Thus, this study aimed to decipher an alternate innate inducer of the expression host by determining the metabolites present in the recombinant strain SO and investigate the expression of the bacterial lipase using the significant selected metabolite (fatty acids). The media from M. guilliermondii strain SO (wild type) and SO2 (recombinant strain) (at 0 and 60 h) were extracted using methanol extraction protocols followed by gas chromatography-mass spectrometry (GC-MS). A multivariate statistical analysis; principle component analysis (PCA) and partial least square discriminant analysis (PLSDA) were implemented to determine the relationship between the metabolites present in strain SO and SO2. The results showed that the primary metabolites; amino acids, organic acids and particularly, copious amounts of fatty acids, were significantly present in strain SO2 compared to strain SO. Further analysis of the identified fatty acids was conducted and the results showed that hexadecanoic acid (C16) showed an increase of 1.45 fold of T1 lipase expression in SO2 compared to the control experiment. This finding suggested that the fatty acid could be used as an alternative inducer for T1 lipase expression to reduce and/or eliminate the application of methanol.
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Page 76 - 85
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Nor Azlan Nor Muhammad, Chong Sheng Cheah, Nor Syafinaz Yaakob
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IN SILICO CHARACTERISATION OF A Polygonum minus SEQUENCE AS PUTATIVE BEACH DOMAIN PROTEIN
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Abstract
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Polygonum minus has been widely studied due to its significant content of flavonoid, phenolic and terpenoid compounds which are well-associated with medicinal properties. However, most of the pathways that regulate the development and synthesis of secondary metabolites of P. minus remained unknown. Identifying candidate genes and proteins that are involved in the biosynthetic pathways would contribute to a better understanding of the bioactive compound synthesis of P. minus. This study analysed and characterised a large 2151 amino acid hypothetical protein in P. minus using bioinformatic tools and databases. Sequence homology search, conserved domain prediction, protein hierarchical clustering, structure prediction and sub-cellular localisation prediction were done. Results from sequence homology search showed that it is a BEACH domain-containing protein. Analysis of its conserved domain revealed the presence of Concanavalin A-like lectin domain (ConA), Pleckstrin homology-like domain (PH), BEACH domain and WD40 domain repeats. Hierarchical clustering of protein supported that it has a close relationship with BEACH protein family members. The structure of the hypothetical protein was proposed and revealed the presence of a small pocket that functions for the binding of dsRNA. Sub-cellular localisation analysis suggested that the hypothetical protein is localized in the endoplasmic reticulum. It is proposed that the hypothetical protein is involved in the autophagy process in P. minus. This study serves as one of the keys to understanding the role of the hypothetical protein in the plant autophagy process which is important in immunity defence and stress tolerance of P. minus. Manipulation of the autophagy process may result in a better yield of secondary metabolites and promote the survival rate of P. minus.
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April 2023
Malay. J. Biochem. Mol. Biol. (2023) 26 (1)
TABLE OF CONTENTS
Page 1- 9
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Anh Cam Ha, Tan Minh Le & Chinh Duc Nguyen Pham
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OPTIMIZATION OF Artocarpus altilis L. EXTRACT FOR XANTHINE OXIDASE INHIBITORY ACTIVITIES USING RESPONSE SURFACE METHODOLOGY (RSM)
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Abstract
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Artocarpus altilis is widely used in traditional medicine, especially in Vietnam. Many studies have reported the bioactivities of A. altilis in anti-diabetes, while its anti-gout activity has not received much interest. In this study, extraction conditions of A. altilis were comprehensively optimized by the response surface methodology (RSM) for the xanthine oxidase inhibitory activity. Furthermore, the effects of the maturity stage on phytochemicals and the xanthine oxidase inhibitory activity of A. altilis leaves were assessed. The results indicated that the old leaves were appropriate materials for anti-xanthine oxidase. Under the optimal conditions at ethanol concentration 79.74 %, time extraction 76.68 min, and solid-to-liquid ratio 1:9.56 g:mL, the value of IC50 in xanthine oxidase inhibitory activity reached 2.67 μg/mL, nine times less than that of crude extract (24.14 μg/mL). The biological activity of the optimal sample also showed the potentiality of inhibition of α-glucosidase enzyme and antioxidant with IC50 values of 64.83 μg/mL and 1.88 μg/mL, respectively.
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Page 10 - 21
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Anjana Chamilka Thuduhenage Dona, Nurul Hammizah Hamidon and Nurhidanatasha Abu Bakar
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A REVIEW OF THE ANTIPARASITIC EFFECTS OF ELLAGIC ACID AND OTHER PHENOLIC COMPOUNDS ISOLATED FROM MEDICINAL PLANTS
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Abstract
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The pursuit of alternative sources for antiparasitic drugs is crucial due to the frequent inadequacy of current treatments and the potential development of resistance among parasites towards synthetic therapies. Conversely, medicinal plants have garnered prominent attention as they produce various natural compounds with intriguing biological properties that can be beneficial in treating parasitic infections. This article presents a comprehensive overview of past literature regarding the topic. It specifically focuses on the antiparasitic effects of ellagic acid and other phenolic compounds against parasites, such as Plasmodium, Trypanosoma, Leishmania, and Helminths. The findings hope to contribute further understanding of the utilisation of ellagic acid and other phenolic compounds as innovative antiparasitic drugs for effectively managing parasitic infections. It also highlights potential limitations and sets the direction for future research.
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Page 22 - 30
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Elisha Apatewen Akanbong, Alparslan Kadir Devrim, Ali Åženol, Mert Sudagidan
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COMPARISON OF SOME COPPER COMPOUNDS IN TERMS OF THEIR INTERACTIONS WITH DNA USING AGAROSE GEL ELECTROPHORESIS, UV-ABSORBANCE AND FLUORESCENCE SPECTROPHOTOMETRY TECHNIQUES
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Abstract
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In this study, to investigate the interaction of genomic DNA with some copper compounds (CuSO4, CuCO3, and CuCl2) at various concentrations (1000 μM, 500 μM, 250 μM, 125 μM and 62.5 μM); UV-absorbance spectrophotometry, agarose gel electrophoresis, and fluorescence spectrophotometry techniques were used. When the UV-spectrophotometry data were examined within the wavelength range of 220-320 nm, the hyperchromic effect of CuCl2 was evaluated to be proportional to its concentrations. Within the wavelength range, the copper compounds produced their strongest hyperchromic effect on the DNA at 1000 μM. When the fluorescent spectrophotometry data were evaluated, the hypochromic effect of CuCl2 within the wavelength range of 400-700 nm was determined at 1000 μM. CuSO4 showed a hypochromic effect at concentrations above 62.5 μM. Also, it was observed that CuCO3 did not interact with DNA. According to the agarose gel electrophoresis findings, the copper compounds investigated decreased DNA band intensity as their concentrations increased. It was observed that the most significant decrease in band intensity was caused by CuCl2 at 1000 μM. Again, it was observed that the copper compounds did not cause any cleavage in the genomic DNA within the concentration range of 62.5-1000 μM. Consequently, the copper compounds bind to DNA, most probably by non-intercalative mode. In this regard, they could have the potential to be used in the development of new therapeutic agents. Therefore, conducting further studies on the interaction of the copper compounds with DNA, their possible DNA uncoiling activities, and also the investigation of these compounds in cancer cell lines will provide useful results.
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Page 31 - 41
Syubbanul Wathon, Ratis Nour Sholichah, Rike Oktarianti, Kartika Senjarini
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REDESIGNING PRIMER OF CYTOCHROME OXIDASE SUBUNIT 1 (CO1) GENE FOR SPECIFIC IDENTIFICATION OF MALARIA VECTOR Anopheles sp.
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Abstract
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Malaria remains a serious global health issue. Due to the absence of a malaria vaccine, vector control is deemed one of the most effective preventive strategies. Anopheles sp., with 20 species, is a vector of malaria in endemic areas across Indonesia. Identifying the vector of Anopheles sp. helps determine an effective and efficient vector control strategy. It is also important to determine disease epidemiology because different species have distinctive characteristics as malaria vectors. Several siblings and cryptic species are difficult to identify based on their morphological characteristics. These limitations have prompted the development of methods for species identification through DNA barcoding techniques. One of the molecular markers widely used in DNA barcoding is the Cytochrome Oxidase Subunit 1 (CO1) gene derived from mitochondrial DNA. This study aimed to redesign the CO1 primer with a database from Anopheles sp. for a more precise and accurate interpretation of Anopheles sp. The design of CO1 primer was developed through a bioinformatics approach using the Anopheles sp. in NCBI database system. The redesigned primer was applied to the genomic DNA sample of Anopheles sp. obtained from the landing collection which had been morphologically identified in advance. The molecular identification on the new CO1 primer was done by CO1 sequence amplification, purification of PCR results, sequencing, and data analysis. The redesigned CO1 primer was named sra-CO1 with a forward sequence of 5' CCCGGAGCATTTATTGGGGA 3' and a reverse sequence of 5' AACCTGTTCCTGCCCCATTT 3' with a product length of 247 bp. The sra-CO1 primer has been successfully used in the molecular identification of Anopheles vagus and Anopheles vagus limosus, with the accession number NCBI Anopheles vagus MK628547.1. These results prove that the sra-CO1 primer can be used in the molecular identification of the genus Anopheles sp.
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Page 42 - 50
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Mariyam Shabeea Ahmed, Siti Aqlima Ahmad, Mohd Yunus Shukor and Mohd Termizi Yusof
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EFFECT OF ACRYLAMIDE ON THE DEGRADATION OF PALM-BASED USED COOKING OIL BY SINGLE AND CO-CULTURE ISOLATES OF Alcaligenes sp. AQ5-02 AND Serratia sp. AQ5-03
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Abstract
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Acrylamide is found in the environment, food, and waste products such as palm-based used cooking oil (UCO). The presence of acrylamide is a threat due to its neurotoxic, genotoxic, teratogenic, and carcinogenic characteristics. These negative impacts have sparked an interest in microbe-mediated bioremediation of acrylamide. The aim of this study is to investigate the effect of various concentrations of acrylamide on the degradation of UCO as a sole carbon source by single and co-culture isolates of Alcaligenes sp. AQ5-02 and Serratia sp. AQ5-03. Using gravimetric analysis, different acrylamide concentrations ranging from 10-200 mg/L were shown to significantly reduce palm oil UCO degradation and bacterial growth in single and co-culture formulations. In the co-culture, there were significant differences between all acrylamide concentrations in terms of UCO degradation. The single and co-culture isolates were able to withstand 10 mg/L of acrylamide while degrading more than 50% of UCO. However, beyond 10 mg/L, there was a significant reduction in the degradation of UCO in both single and co-culture methods. Two-way ANOVA reveals a significant difference in the degradation and bacterial growth between the single and co-culture isolates of Alcaligenes sp. AQ5-02 and Serratia sp. AQ5-03 with varying acrylamide concentrations (p < 0.0001). In conclusion, the co-culture has greater degradation and better tolerance at all concentrations of acrylamide compared to single isolate cultures of Alcaligenes sp. AQ5-02 and Serratia sp. AQ5-03. The study provides insightful knowledge on the presence of xenobiotics in the bioremediation of hydrocarbons by single and co-culture.
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Page 51 - 58
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Fathmath Shaman Fareed, Nallammai Singaram
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A MODIFIED RNA EXTRACTION PROTOCOL FOR SECONDARY METABOLITE-RICH AMOMUM SPECIES (ZINGIBERACEAE)
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Abstract
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High-quality and pure RNA are necessary for transcriptomic studies using next-generation sequencing technologies (NGS). Peninsular Malaysian ginger species are well known for their therapeutic properties and high levels of secondary metabolites such as alkaloids, flavonoids, and aromatic compounds. The Amomum genus of this family is particularly well-known for its significant antioxidant and antimicrobial properties. Although many plant RNA extraction protocols have been developed in the past, the efficacy varies depending on the species and the plant and plant parts utilized. Therefore, this study aimed to compare different RNA extraction protocols and determine a protocol for the secondary-metabolite-rich ginger species of the Zingiberaceae family. RNA was extracted from fresh leaves, stems, roots, and rhizomes of Amomum genus using four different methods: a commercial kit, CTAB-LiCl, SDS-LiCl and SDS-Phenol. Based on the findings, the modified commercial kit produced high-quality, pure RNA for each plant tissue utilized, surpassing the other approaches. The findings of this study emphasize the importance of PVP, and β-ME in the extraction process, particularly in Zingiberaceae species with high secondary metabolites.
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Page 59 - 65
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Nor Azlianie A@A, Zunoliza A, Nurhanan MY, Nur Vicky B, Nor Jannah S and Mohd Hafidz Hadi A
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ANTI-PROLIFERATIVE ACTIVITIES OF Tectona grandis LEAVES AGAINST SELECTED CANCER CELL LINES
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Abstract
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Tectona grandis Linn (Teak), known locally as Sagwan, is a member of the Verbenaceae family. The leaves of this plant have been reported to contain various groups of compounds including anthraquinones, lignin derivatives, anthratectone and naphthatectone which are made up of several compounds such as verbascoside, isoverbascoside, abeograndinoic acid, tectoquinone, lapachol and deoxylapachol. Pharmacologically, the plant leaves have been studied for in vitro anticancer, antibacterial, antifungal, antiviral, antiprotozoal, insecticidal, anti-inflammatory and antipyretic properties. In this study, the methanol extract (TGM) of T. grandis leaves was subjected to tannin removal to yield 5 fractions consisting of TGF1 (100% water), TGF2 (50% methanol), TGF3 (100% methanol), TGF4 (5% acetic acid) and TGF5 (0.1N NaOH). The extract and fractions were assessed using the sulforhodamine B assay for anti-proliferative activities against ovarian, breast and colorectal cancer cell lines. The TGM extract showed moderately active anti-proliferative activities against A2780 (IC50 34.38 ± 0.94 μg/ml), SKOV-3 (IC50 37.73 ± 1.11 μg/ml), MCF-7 (IC50 50.94 ± 0.78 μg/ml) and HT-29 (IC50 49.67 ± 0.78 μg/ml) cancer cell lines. TGF4 fraction also showed moderately active anti-proliferative activity against A2780 (IC50 25.21 ± 0.52 μg/ml), SKOV-3 (IC50 33.49 ± 0.074 μg/ml), MCF-7 (IC50 32.80 ± 0.17 μg/ml) and HT-29 (IC50 33.05 ± 0.20 μg/ml) cancer cell lines as compared to other fractions, TGF1, TGF2, TGF3 and TGF5, which were not active. The chemical compounds present in the extract and fractions obtained were characterized and identified by chromatography analysis which are caffeic acid derivatives and flavonoid compounds.
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Page 66 - 76
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Nur Farhani Binti Mohd Ghani, Gurmeet Kaur Surindar Singh, Nurul Aqmar Mohd Nor Hazalin
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NEUROPATHOLOGY OF STREPTOZOTOCIN-INDUCED RODENT MODELS OF ALZHEIMER’S DISEASE: A REVIEW ON BEHAVIORAL AND HISTOLOGICAL EVIDENCE
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Abstract
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Alzheimer's disease (AD) is an incurable neurodegenerative disease with significant research efforts focused on developing effective treatments. Various rodent models, including transgenic and non-transgenic models, have been established to investigate AD and potential therapeutic interventions. Streptozotocin (STZ), a naturally alkylating antineoplastic agent with a diabetogenic effect on mammals is the widely used sporadic rodent AD model due to its ability to mimic certain aspects of sporadic AD observed in humans. Recent evidence has highlighted a correlation between STZ administration and AD-like neuropathology, characterized by exacerbated neuroinflammation and the manifestation of AD hallmarks in animals. However, certain key characteristics of STZ-induced AD pathology remain poorly described. Therefore, this review aims to summarize the neuropathological hallmarks of AD in rodents following STZ administration. STZ injection chronically produces multiple effects resembling AD's behavioural and pathological aspects. Rodents that received injections of STZ developed long-term progressive deficits of memory, learning and cognitive behaviour. Histologically, STZ affects neurons and synapses in the brain, accompanied by the presence of amyloid-beta (Aβ) plaques, tau hyperphosphorylation, white matter atrophy, and myelin damage. Understanding the connection between behavioural and neuropathological alterations following STZ administration and their relevance to AD pathology in rodents would significantly contribute to the field of AD animal models.
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Page 77 - 82
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Nur Azizah Nadhirah Binti Azni, Yong Qi Leong, Chooi Ling Lim, Khuen Yen Ng, Soi Moi Chye, Anna Pick Kiong Ling, Yin Yin Ooi, Rhun Yian Koh
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THE ROLES OF ANTIFIBROTIC THERAPIES IN POST-SARS-CoV-2 INFECTION
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Abstract
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Coronavirus Disease 2019 (COVID-19) pandemic, caused by the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), has affected over a hundred million people worldwide. Other than acute symptoms after infection, the patients are also suffering from long-term health complications associated with COVID-19 called long COVID or post-COVID conditions by which pulmonary fibrosis (PF) was observed in one-third of the patients. Since post-COVID-19-associated PF (PCPF) shares similar symptoms with idiopathic pulmonary fibrosis (IPF), the ongoing studies focus on the potential use of therapeutic approaches for IPF in PCPF patients. Hence, this study summarises the potential roles of these therapies to treat the PCPF including antifibrotic drugs, mesenchymal stem cell (MSC) therapy and inhaled curcumin nanoformulations by assessing their efficacies for the PCPF treatment.
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Page 83 - 89
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Sri Widyarti, Bayu Dwi Prakoso, Shifaaun Najihah, Syahputra Wibowo, Zakia Hasna Fajar, Sofy Permana, Sutiman Bambang Sumitro
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PHYSICOCHEMICAL CHARACTERIZATION OF ASTAXANTHIN-Cu COMPLEXES
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Abstract
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Astaxanthin (ASX) is a natural biological antioxidant acceptable as a dietary supplement and food colorant. ASX has unique molecular characteristics which form stable formations in the presence of metal ions. This study aims to analyze the astaxanthin-Cu complex using FTIR, SEM-EDS, and XRD. The synthesis of the complex carried on ratios 1:1, 1:2, and 2:1. Complex formation was determined using UV-Vis spectrum and then confirmed using FTIR. Although the profile spectrum UV-Vis has no differences between ratios, FTIR analysis showed an interaction between ASX with Cu. The SEM-EDS microstructure analysis showed that the ASX-Cu complex is a more regular form than astaxanthin alone. The analysis results based on X-ray diffraction showed that the crystal of astaxanthin, Cu, and each complex had different structures, asymmetric, and atomic arrangements. In conclusion, the ASX-Cu complex with a ratio of 1:1, 1:2, and 2:1 has been successfully made at the incubation time of 5 minutes at 20 degrees C.
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Page 90 - 97
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Aliyya Suci Arizona, Wira Eka Putra, Hendra Susanto, Sustiprijatno, Arief Hidayatullah, Muhammad Fikri Heikal, Diana Widiastuti
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IN SILICO INVESTIGATION OF MUNG BEAN (Vigna radiata L.) ACTIVE COMPOUNDS AS POTENTIAL NATURAL INHIBITOR AGAINST HEPATITIS C VIRUS E2 ENVELOPE GLYCOPROTEIN
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Abstract
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The hepatitis C virus (HCV) is the causative agent of hepatitis C, an infection of the liver. The liver may suffer long-term damage as a result of this illness. A significant number of people in Indonesia are infected with hepatitis C. Because of the restrictions and potential side effects of their hepatitis C treatment, a large number of people decide to stop receiving medication. Traditional remedies made from plants have started to become more popular in recent years due to the fact that they are less expensive and have fewer adverse effects. Thus, in this present study, we aimed to evaluate the Vigna radiata active constituent as a possible anti-viral drug against Hepatitis C Virus Envelope E2 Glycoprotein through in silico approaches. The Lipinski rule of five is used in the process of virtually screening compounds to identify those that have properties similar to candidates of the drug. Following that, the target chemicals and proteins were adjusted in preparation for additional molecular docking. In order to get binding affinity scores, chemical interactions, ligand sites, and amino acid residues, visualization and data analysis were carried out. Based on our investigation, we found the top five molecules with lower binding energy values than IFN-α as control drugs including isovitexin, orientin, vitexin, caffeic acid, and p-coumaric acid. In addition, the findings demonstrated that the chemical bonds that facilitate the contact process are present in every substance that forms a binding interaction with the target protein. As a result, our findings provide preliminary evidence that the V. radiata plant has the potential to act as an anti-viral medicine that is effective against hepatitis C virus E2 envelope glycoprotein.
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Page 98 - 112
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Muhammad Syahmi Abd Rahman, Zixin Hong and Nadiya Akmal Baharum
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IN SILICO CHARACTERIZATION OF DEFENSIN IN Musa acuminata DH PAHANG (MaDef) PROVIDES INSIGHT INTO POTENTIAL DEFENCE-RELATED PROTEIN
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Abstract
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Plant defensins are expressed in response to phytopathogens and various defence-related signalling molecules. They possess diverse biological properties such as antifungal, antibacterial and proteinase inhibitory, as well as playing roles in plant growth and development. Multiple defensin copies are identified in numerous plant species, such as Arabidopsis, Brassica oleracea, Zea mays and Medicago truncatula. To our knowledge, the multigene family of defensin has never been reported in bananas. In addition, specific banana defensin genes involved in the defence and stress responsiveness are yet to be identified. Thus, this study predicts specific copies of Musa acuminata DH Pahang (wild banana) defensins that are potentially involved with defence and biotic stress response using in silico analysis. A total of 6 defensin copies from Musa acuminata DH Pahang (wild banana) (MaDef) were identified and categorised under the Knottin_1 clan (CL0054). All of them except Ma07_t03680.1 carry conserved sequences of the gamma-thionin domain (PF00304). A total of 8 cysteines forming 4 disulfide bridges are found across all six MaDef peptide sequences. Using phylogenetic analysis, wild banana defensins are categorised under three clades with a predicted molecular weight of 8 to 9 kDa. Gene ontology (GO) revealed that all MaDefs except for Ma07_t03680.1 are involved in defence response. Furthermore, analysis of the promoter regions through PlantCARE shows Ma04_t36140.1 is associated with defence and stress responsiveness. Overall, this study contributes to a deeper understanding of defensins characteristics and functional predictions, which are critical for future crop advances against biotic challenges, notably in bananas.
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Page 113 - 121
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Sien-Yei Liew, Noraini Abd-Aziz, Eric J Stanbridge, Khatijah Yusoff, Norazizah Shafee, Suhaili Mustafa
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ESTABLISHMENT OF A HIGHLY SENSITIVE CELL-BASED ASSAY FOR SCREENING OF HYPOXIA-INDUCIBLE FACTOR REGULATORS
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Abstract
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Cell-based assay is a powerful tool in the field of drug discovery. One of the main targets of drug development is transcription factors. Hypoxia-inducible factors (HIFs) are transcription factors involved in cellular oxygen homeostasis and associated with disease pathophysiology. Currently, a highly sensitive cell-based assay to measure HIF activity is still lacking. Using luciferase reporter constructs, we developed a highly sensitive, stable reporter cell line for monitoring the activity of HIF. Four copies of hypoxia response element (HRE) of the erythropoietin (EPO) gene, which is one of the targets of HIF, were used to drive the expression of the luciferase reporter. This stable reporter cell line gave a 500-fold increase in the luciferase signal in hypoxia compared to a normoxia condition. This robust increase in hypoxia response is crucial to ensure consistency and reproducibility of HIF assay results while reducing the overall assay cost. The establishment of this highly sensitive cell-based HIF assay may help expedite research in the discovery of HIF regulatory drugs.
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Page 122 - 134
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Tran Thi Phuong Nhung, Le Pham Tan Quoc
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ASSESSMENT OF TOXIC EFFECTS OF Hedyotis capitellata WALL. LEAVES ETHANOL EXTRACT VIA BIOLOGICAL ASSAYS IN MICE (Mus musculus)
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Abstract
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Hedyotis capitellata Wall. is a traditional plant known for its various significant pharmacological effects, such as gastric protection, pain relief, anti-inflammatory properties, etc. Despite its wide usage in traditional medicine, no scientific studies have been published on its toxicity. The current study evaluated the potential toxicity of ethanol leaf extract of H. capitellata (EtHC) through acute and sub-chronic oral administration in mice following guidelines 423 and 408 of the OECD for chemical testing. To assess the acute toxicity of EtHC, single doses of 1000, 3000, and 5000 mg/kg body weight were administered to mice and continuously observed for 14 days. Additionally, daily doses of 100, 300, and 500 mg/kg body weight were also given to mice for 90 days to investigate sub-chronic toxicity. At the end of the study, blood, urine, and vital organs were collected for haematological, biochemical, urine analysis, and histopathological studies. The results demonstrated no cases of mortality or signs of toxicity observed in the acute toxicity test. Throughout the 90-day study period, all tested extract doses exhibited no harmful effects on the organs. Haematological and biochemical parameters in the acute toxicity (5000 mg/kg) and subchronic toxicity (500 mg/kg) groups (e.g., red blood cell count: 6.59 ± 0.41 × 106 cells/mm3, aspartate aminotransferase (AST) level: 16.62 ± 0.31 U/L, alanine aminotransferase (ALT) level: 18.15 ± 0.58 U/L, urine (pH 7.05 ± 0.09, specific gravity 1.17 ± 0.11) did not significantly differ from the control group (p > 0.05). However, individual differences were observed in the white blood cell count (EtHC5000 group) and glucose level (EtHC500 group) compared to the control group (p < 0.05). These changes were considered individual variations and not biologically significant. Histopathological examination of the organs also revealed no abnormalities in the extract-treated group. This study demonstrates that the ethanol leaf extract of H. capitellata is non-toxic and could be considered for treating various diseases, such as pain relief, anti-inflammatory effects, gastric ulcer treatment, etc.
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Page 135 - 151
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Nur Athirah Zabidi, Muhajir Hamid, Mohd. Ezuan Khayat, Mohd Badrin Hanizam Abdul Rahim, Shafinaz Abd Gani
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TOWARDS A NOVEL BIO-INSPIRED DRUG DELIVERY SYSTEM: EXOSOMES LOADED WITH POLYPHENOLS AS NATURAL DRUG CARRIERS IN DIABETES TREATMENT
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Abstract
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Polyphenols have been used as active ingredients in pharmaceuticals, healthcare goods, and dietary supplements to improve the functional properties of food. They have been widely investigated as an antidiabetic agent in vitro and in vivo studies. Polyphenols are known to exert antidiabetic properties as a potent inhibitor or as an activator to minimize β-cell dysfunction and insulin resistance in Diabetes Mellitus (DM). Nevertheless, due to some impaired characteristics such as low bioavailability, rapid elimination from the body, high-rate metabolisms, poor absorption due to hydrophobic properties, and less stability, these compounds are prevented from performing their optimal therapeutic activities. Exosomes, nanoparticles made from different types of cells, have a great deal of potential to be one of the promising carriers in drug delivery systems to address these problems (DDS). To date, exosome exhibits particular benefits as a natural drug delivery agent in terms of specificity, stability, and safety that mediate the synergistic effect between polyphenols and diabetes. Exosomes have recently been used as a novel medication delivery method, increasing the effectiveness and efficiency of the drug delivery system. According to certain studies, exosome-nano encapsulating polyphenols could improve their biological effect in terms of bioavailability and bioaccessability. In addition, accumulated evidence illustrates that nanoencapsulation of polyphenols with exosomes enhances metabolic pathways in DM progression. In this review, we explore the effect of polyphenols encapsulated with exosomes concerning their beneficial effect on DM management.
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Page 152 - 162
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Vi Sion Chang, Yong Hui Wong, Kumar Veerapen, Zoe Yi Ng, Wei Min Hon, Renee Lay Hong Lim
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ASSOCIATION OF TGFB1 rs1800470 AND TGIF rs2229333 VARIANTS WITH MYOPIA IN MALAYSIAN ADOLESCENTS
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Abstract
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Myopia is a global disease with high prevalence in school children. This study aimed to investigate the prevalence of myopia and its association with variants TGFB1 rs1800470 and TGIF rs2229333 among Malaysian adolescents. A total of 565 Malaysian adolescents (206 Chinese, 220 Malay, and 139 Indian) were recruited and refractive error measurements were used for grouping subjects to normal or myopes (≤-0.50D). Concurrently, genomic DNA from buccal cells was genotyped using conventional PCR-RFLP. Allelic and genotype frequency in association with myopia prevalence were statistically analysed. High myopia prevalence (83.0%) was detected in the study population, with females (85.8%) being more myopic than males (78.8%). The highest prevalence of myopia was observed in Chinese (87.4%) followed by Malay (82.3%) and Indian (77.7%) (p<0.001) adolescents. In TGFB1, T allele was the minor allele in the Chinese and Malays but not Indians, whereas in TGIF, T was the minor allele in all ethnic groups. Interestingly, myopes compared to normal showed higher frequency for CT or combined (CT+CC) genotype for TGFB1, but only Malay males showed significantly higher CT (p=0.033, relative risk=1.23) and (CT+CC) (p=0.009, OR (95% CI) = 4.23) respectively. Despite the higher frequency of combined (CT+TT) genotype for T risk allele of TGIF observed for myopes, no significant association with myopia was detected. The genotype and allele frequency of both variants differed based on sex and ethnicity. This is the first study demonstrating a significant association of the TGFB1 variant with myopic Malay male adolescents, and no association between TGIF variant and myopia in this study population.
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