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Optimization of total phenolic content, total flavonoid content and anti-gout properties of polyherbal formulation

Yee LS, Abu Bakar MF, Abdullah N, Abu Bakar FI, Fatmawati S

J Complement Integr Med, 2023 Dec 01;20(4):772-778.
PMID: 37561949 DOI: 10.1515/jcim-2020-0141

OBJECTIVES: An increase in gout prevalence has drawn attention among society and this situation drives the exploration of more favourable treatment using traditional medicinal plants which are rich in phenolic and flavonoid to avoid the side effects of modern medication. However, there are only few studies regarding the optimization of phytochemicals and anti-gout properties of medicinal plants and their combinations. The objectives of this study were to determine the optimal formulation of Strobilanthes crispus, Orthosiphon stamineus Benth and Stevia rebaudiana with maximum total phenolic and flavonoid contents as well as minimum IC50 of in vitro xanthine oxidase inhibitory activity and to examine their correlations among the formulations.
METHODS: Plant extracts from hot water infusion were tested for the total phenolic content, total flavonoid content and enzyme inhibition through Folin-ciocalteu assay, aluminium chloride method and xanthine oxidase inhibition assay, respectively. Simplex-centroid mixture design was applied in this study and 13 polyherbal formulations were generated by Design Expert Software.
RESULTS: Linear, special cubic and quadratic models were selected to describe the interaction effect between polyherbal formulations and their responses. Low IC50 value (13.90 μg/mL) of xanthine oxidase activity was found in the binary combination of O. stamineus and S. rebaudiana and this probably related to its high phenolic and flavonoid contents as xanthine oxidase inhibition and phytochemicals were correlated.
CONCLUSIONS: The suggested optimal formulation was comprised of 44.26 % O. stamineus and 55.74 % S. rebaudiana and it could be developed as an alternative treatment for gout.
Fulltext Phytochemical Constituents, Antioxidant and Antiproliferative Properties of a Liverwort, Lepidozia borneensis Stephani from Mount Kinabalu, Sabah, Malaysia

Abu Bakar MF, Abdul Karim F, Suleiman M, Isha A, Rahmat A

Evid Based Complement Alternat Med, 2015;2015:936215.
PMID: 26640502 DOI: 10.1155/2015/936215

The study aimed to investigate the phytochemical contents, antioxidant and antiproliferative activity of 80% methanol extract of Lepidozia borneensis. The total phenolic and total flavonoid contents were analysed using Folin-Ciocalteu and aluminium chloride colorimetric methods. Antioxidant properties were evaluated by using FRAP, ABTS, and DPPH assays while the effects of L. borneensis on the proliferation of MCF-7 cell line were evaluated by using MTT assay. The results showed that the total phenolic and flavonoid contents were 12.42 ± 0.47 mg GAE/g and 9.36 ± 1.29 mg CE/g, respectively. The GC-MS analysis revealed the presence of at least 35 compounds. The extract was found to induce cytotoxicity against MCF-7 cell line with IC50 value of 47.33 ± 7.37 µg/mL. Cell cycle analysis showed that the extract induced significant arrest at G0/G1 at 24 hours of treatment. After 72 hours of treatment, the proportion of cells in G0/G1 and G2-M phases had decreased significantly as compared to their control. Apoptosis occurred during the first 24 hours and significantly increased to 30.8% after 72 hours of treatment. No activation of caspase 3 was observed. These findings suggest that L. borneensis extract has the potential as natural antioxidant and anticancer agents.
Fulltext Secondary Metabolites, Antioxidant, and Antiproliferative Activities of Dioscorea bulbifera Leaf Collected from Endau Rompin, Johor, Malaysia

Mainasara MM, Abu Bakar MF, Md Akim A, Linatoc AC, Abu Bakar FI, Ranneh YKH

Evid Based Complement Alternat Med, 2021;2021:8826986.
PMID: 33505506 DOI: 10.1155/2021/8826986

Breast cancer is among the most commonly diagnosed cancer and the leading cause of cancer-related death among women globally. Malaysia is a country that is rich in medicinal plant species. Hence, this research aims to explore the secondary metabolites, antioxidant, and antiproliferative activities of Dioscorea bulbifera leaf collected from Endau Rompin, Johor, Malaysia. Antioxidant activity was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assays, while the cytotoxicity of D. bulbifera on MDA-MB-231 and MCF-7 breast cancer cell lines was tested using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Cell cycle analysis and apoptosis were assessed using flow cytometry analysis. Phytochemical profiling was conducted using gas chromatography-mass spectrometry (GC-MS). Results showed that methanol extract had the highest antioxidant activity in DPPH, FRAP, and ABTS assays, followed by ethyl acetate and hexane extracts. D. bulbifera tested against MDA-MB-231 and MCF-7 cell lines showed a pronounced cytotoxic effect with IC50 values of 8.96 μg/mL, 6.88 μg/mL, and 3.27 μg/mL in MCF-7 and 14.29 μg/mL, 11.86 μg/mL, and 7.23 μg/mL in MDA-MB-231, respectively. Cell cycle analysis also indicated that D. bulbifera prompted apoptosis at various stages, and a significant decrease in viable cells was detected within 24 h and substantially improved after 48 h and 72 h of treatment. Phytochemical profiling of methanol extract revealed the presence of 39 metabolites such as acetic acid, n-hexadecanoic acid, acetin, hexadecanoate, 7-tetradecenal, phytol, octadecanoic acid, cholesterol, palmitic acid, and linolenate. Hence, these findings concluded that D. bulbifera extract has promising anticancer and natural antioxidant agents. However, further study is needed to isolate the bioactive compounds and validate the effectiveness of this extract in the In in vivo model.
Fulltext Southeast Asian Medicinal Plants as a Potential Source of Antituberculosis Agent

Sanusi SB, Abu Bakar MF, Mohamed M, Sabran SF, Mainasara MM

Evid Based Complement Alternat Med, 2017;2017:7185649.
PMID: 29081822 DOI: 10.1155/2017/7185649

Despite all of the control strategies, tuberculosis (TB) is still a major cause of death globally and one-third of the world's population is infected with TB. The drugs used for TB treatment have drawbacks of causing adverse side effects and emergence of resistance strains. Plant-derived medicines have since been used in traditional medical system for the treatment of numerous ailments worldwide. There were nine major review publications on antimycobacteria from plants in the last 17 years. However, none is focused on Southeast Asian medicinal plants. Hence, this review is aimed at highlighting the medicinal plants of Southeast Asian origin evaluated for anti-TB. This review is based on literatures published in various electronic database. A total of 132 plants species representing 45 families and 107 genera were reviewed; 27 species representing 20.5% exhibited most significant in vitro anti-TB activity (crude extracts and/or bioactive compounds 0-<10 µg/ml). The findings may motivate various scientists to undertake the project that may result in the development of crude extract that will be consumed as complementary or alternative TB drug or as potential bioactive compounds for the development of novel anti-TB drug.
Fulltext Optimization of Extraction Conditions of Phytochemical Compounds and Anti-Gout Activity of Euphorbia hirta L. (Ara Tanah) Using Response Surface Methodology and Liquid Chromatography-Mass Spectrometry (LC-MS) Analysis

Abu Bakar FI, Abu Bakar MF, Abdullah N, Endrini S, Fatmawati S

Evid Based Complement Alternat Med, 2020;2020:4501261.
PMID: 32047524 DOI: 10.1155/2020/4501261

Gout is a common disease affected most of the people due to the elevation of uric acid in the blood. Flavonoid and phenolic compounds are reported to exert the anti-gout activity of medicinal plants. Hence, this study aimed at optimizing the extraction conditions of phenolic and flavonoid compounds as well as the anti-gout (xanthine oxidase inhibitory activity) in vitro of Euphorbia hirta using response surface methodology (RSM). The plant part used was the whole plant excluding roots. The effects of three independent variables (extraction time, X1; extraction temperature, X2; and solid-to-liquid ratio, X3) on three response variables (total flavonoid content, Y1; total phenolic content, Y2; and xanthine oxidase inhibitory activity, Y3) were determined using central composite design (CCD) while phytochemical profiling of the extracts was determined by liquid chromatography-mass spectrometry (LC-MS). Quadratic models produced a satisfactory fitting of the experimental data with regard to total flavonoid content (r2 = 0.9407, p < 0.0001), total phenolic content (r2 = 0.9383, p < 0.0001), and xanthine oxidase inhibitory activity (r2 = 0.9794, p < 0.0001). The best extraction conditions observed for total flavonoid content, total phenolic content, and xanthine oxidase inhibitory activity were at a temperature of 79.07°C for 17.42 min with solid-to-liquid ratio of 1 : 20 g/ml. The optimum values for total flavonoid, total phenolic, and xanthine oxidase inhibitory activity were 67.56 mg RE/g, 155.21 mg GAE/g, and 91.42%, respectively. The main phytochemical compounds in the optimized E. hirta extract are neochlorogenic acid, quercetin-3β-D-glucoside, syringic acid, caffeic acid, ellagic acid, astragalin, afzelin, and quercetin. As conclusion, this study clearly demonstrated the best conditions to obtain higher xanthine oxidase inhibitory activity and phytochemical compounds which can be further used for the development of anti-gout agents.
Fulltext Phytochemicals from Mangifera pajang Kosterm and their biological activities

Ahmad S, Sukari MA, Ismail N, Ismail IS, Abdul AB, Abu Bakar MF, et al.

BMC Complement Altern Med, 2015;15:83.
PMID: 25887035 DOI: 10.1186/s12906-015-0594-7

Mangifera pajang Kosterm is a plant species from the mango family (Anacardiaceae). The fruits are edible and have been reported to have high antioxidant content. However, the detailed phytochemical studies of the plant have not been reported previously. This study investigates the phytochemicals and biological activities of different parts of Mangifera pajang.
Fulltext Genome Sequence ofSerratia marcescenssubsp.sakuensisStrain K27, a Marine Bacterium Isolated from Sponge (Haliclona amboinensis)

Cheng TH, Saidin J, Danish-Daniel M, Gan HM, Mat Isa MN, Abu Bakar MF, et al.

Genome Announc, 2018 Feb 08;6(6).
PMID: 29439033 DOI: 10.1128/genomeA.00022-18

Serratia marcescens
subsp.sakuensisstrain K27 was isolated from sponge (Haliclona amboinensis). The genome of this strain consists of 5,325,727 bp, with 5,140 open reading frames (ORFs), 3 rRNAs, and 67 tRNAs. It contains genes for the production of amylases, lipases, and proteases. Gene clusters for the biosynthesis of nonribosomal peptides and thiopeptide were also identified.
Fulltext Vitamin D deficiency in Malaysian adolescents aged 13 years: findings from the Malaysian Health and Adolescents Longitudinal Research Team study (MyHeARTs)

Al-Sadat N, Majid HA, Sim PY, Su TT, Dahlui M, Abu Bakar MF, et al.

BMJ Open, 2016 08 18;6(8):e010689.
PMID: 27540095 DOI: 10.1136/bmjopen-2015-010689

OBJECTIVE: To determine the prevalence of vitamin D deficiency (<37.5 nmol/L) among young adolescents in Malaysia and its association with demographic characteristics, anthropometric measures and physical activity.
DESIGN: This is a cross-sectional study among Form 1 (year 7) students from 15 schools selected using a stratified random sampling design. Information regarding sociodemographic characteristics, clinical data and environmental factors was collected and blood samples were taken for total vitamin D. Descriptive and multivariable logistic regression was performed on the data.
SETTING: National secondary schools in Peninsular Malaysia.
PARTICIPANTS: 1361 students (mean age 12.9±0.3 years) (61.4% girls) completed the consent forms and participated in this study. Students with a chronic health condition and/or who could not understand the questionnaires due to lack of literacy were excluded.
MAIN OUTCOME MEASURES: Vitamin D status was determined through measurement of sera 25-hydroxyvitamin D (25(OH)D). Body mass index (BMI) was classified according to International Obesity Task Force (IOTF) criteria. Self-reported physical activity levels were assessed using the validated Malay version of the Physical Activity Questionnaire for Older Children (PAQ-C).
RESULTS: Deficiency in vitamin D was seen in 78.9% of the participants. The deficiency was significantly higher in girls (92.6%, p<0.001), Indian adolescents (88.6%, p<0.001) and urban-living adolescents (88.8%, p<0.001). Females (OR=8.98; 95% CI 6.48 to 12.45), adolescents with wider waist circumference (OR=2.64; 95% CI 1.65 to 4.25) and in urban areas had higher risks (OR=3.57; 95% CI 2.54 to 5.02) of being vitamin D deficient.
CONCLUSIONS: The study shows a high prevalence of vitamin D deficiency among young adolescents. Main risk factors are gender, ethnicity, place of residence and obesity.
Unravelling the adaptation strategies employed by Glaciozyma antarctica PI12 on Antarctic sea ice

Bharudin I, Abu Bakar MF, Hashim NHF, Mat Isa MN, Alias H, Firdaus-Raih M, et al.

Mar Environ Res, 2018 Jun;137:169-176.
PMID: 29598997 DOI: 10.1016/j.marenvres.2018.03.007

Glaciozyma antarctica PI12, is a psychrophilic yeast isolated from Antarctic sea. In this work, Expressed Sequence Tags (EST) from cells exposed to three different temperatures; 15 °C, 0 °C and -12 °C were generated to identify genes associated with cold adaptation. A total of 5376 clones from each library were randomly picked and sequenced. Comparative analyses from the resulting ESTs in each condition identified several groups of genes required for cold adaptation. Additionally, 319 unique transcripts that encoded uncharacterised functions were identified in the -12 °C library and are currently unique to G. antarctica. Gene expression analysis using RT-qPCR revealed two of the unknown genes to be up-regulated at -12 °C compared to 0 °C and 15 °C. These findings further contribute to the collective knowledge into G. antarctica cold adaptation and as a resource for understanding the ecological and physiological tolerance of psychrophilic microbes in general.
Fulltext De novo transcriptome resources of the lichens, Dirinaria sp. UKM-J1 and UKM-K1 collected from Jerantut and Klang, Malaysia

Bharudin I, Abdul Rahim SN, Abu Bakar MF, Ibrahim SN, Kamaruddin S, Latif MT, et al.

Data Brief, 2018 Aug;19:2416-2419.
PMID: 30229114 DOI: 10.1016/j.dib.2018.07.020

Lichen is a symbiotic organism that exists as a single composite body consisting of a mycobiont (fungus) and a photobiont (algae or a cyanobacterium). Many lichen species are considered as extremophiles due to their tolerance to radiation, desiccation, temperature and pollution. However, not all lichen species are tolerant to harsh environmental conditions as several species are sensitive for example to nitrogen, sulphur, acidity, heavy metals, halogens (e.g. fluoride) and ozone. Thus, to better understand why some lichens can withstand exposure to pollutants as opposed to those that are susceptible, we focused on the lichen species of Dirinaria known for their wide distribution in the tropics, subtropics and pantropical, and moderate tolerance to air pollution. Their moderate tolerance to air pollution affords them to thrive in good air quality environments as well as polluted air environments. Lichen samples of Dirinaria sp., UKM-J1 and UKM-K1, were respectively collected from two areas with different levels of air quality based on Air Pollutant Index or API (with index pollutant criteria of PM10, carbon monoxide, ozone, nitrogen dioxide and sulfur dioxide) in the outskirt of Jerantut (UKM-J1), a rural area in the middle of Peninsular Malaysia and the township of Klang (UKM-K1), in a busy area of the Klang Valley, Malaysia. API was monitored throughout 2012-2013 whereby the sample collection site in Klang showed markedly higher concentrations of pollutants in all the index pollutant criteria as compared to that of Jerantut. We performed transcriptome sequencing using Illumina RNA-seq technology and de novo assembly of the transcripts from the lichen samples. Raw reads from both libraries were deposited in the NCBI database with the accession number SRP138994.
Fulltext Transcriptome landscape of Rafflesia cantleyi floral buds reveals insights into the roles of transcription factors and phytohormones in flower development

Amini S, Rosli K, Abu-Bakar MF, Alias H, Mat-Isa MN, Juhari MA, et al.

PLoS One, 2019;14(12):e0226338.
PMID: 31851702 DOI: 10.1371/journal.pone.0226338

Rafflesia possesses unique biological features and known primarily for producing the world's largest and existing as a single flower. However, to date, little is known about key regulators participating in Rafflesia flower development. In order to further understand the molecular mechanism that regulates Rafflesia cantleyi flower development, RNA-seq data from three developmental stages of floral bud, representing the floral organ primordia initiation, floral organ differentiation, and floral bud outgrowth, were analysed. A total of 89,890 transcripts were assembled of which up to 35% could be annotated based on homology search. Advanced transcriptome analysis using K-mean clustering on the differentially expressed genes (DEGs) was able to identify 12 expression clusters that reflect major trends and key transitional states, which correlate to specific developmental stages. Through this, comparative gene expression analysis of different floral bud stages identified various transcription factors related to flower development. The members of WRKY, NAC, bHLH, and MYB families are the most represented among the DEGs, suggesting their important function in flower development. Furthermore, pathway enrichment analysis also revealed DEGs that are involved in various phytohormone signal transduction events such as auxin and auxin transport, cytokinin and gibberellin biosynthesis. Results of this study imply that transcription factors and phytohormone signalling pathways play major role in Rafflesia floral bud development. This study provides an invaluable resource for molecular studies of the flower development process in Rafflesia and other plant species.
Patient profile and antibiotic use in a dedicated upper respiratory tract infection clinic based in a primary healthcare setting during COVID-19 pandemic in Malaysia: A cross sectional study

Ooi ZY, Mohd Ghazali NA, Nik Zahari NJ, Chan HK, Md Noor N, Harun NL, et al.

Malays Fam Physician, 2022 Nov 30;17(3):74-83.
PMID: 36606160 DOI: 10.51866/oa.38

INTRODUCTION: Upper respiratory tract infection (URTI) is commonly encountered at the primary care level. Its management is particularly challenging due to the similarity of its symptoms to coronavirus disease 2019 (COVID-19) infection. Our study evaluated the profiles and antibiotic use of patients seeking care from a dedicated community-based URTI clinic during the COVID-19 pandemic.
METHOD: A cross-sectional study was conducted. Data were obtained from the medical records of patients visiting the URTI clinic at the Alor Setar Primary Healthcare Centre between March and April 2020.
RESULTS: Overall, 587/4388 (13.3%) patients received treatment at the URTI clinic. Most patients were male (60.6%) and aged between 20 and 39 years (35.5%). Their most common symptoms were cough (68.4%), fever (31.6%), runny nose (24.6%), and sore throat (24.1%). Most patients were diagnosed with acute nasopharyngitis (52.5%), acute pharyngitis (18.6%), or acute tonsillitis (5.3%). The symptomatic medication prescription rate was 96.5%. Only 26 of the 435 patients diagnosed with URTI received antibiotics, yielding an antibiotic use rate of only 6.0% for URTI relative to overall drug use. Acute tonsillitis was more common in children <12 years old (p<0.001), while a cough and runny nose were more commonly indicative of acute nasopharyngitis than other conditions (p<0.001). Sore throat was more likely to be a symptom of acute pharyngitis (p<0.001) and acute tonsillitis (p<0.001).
CONCLUSION: Despite the challenges faced during the COVID-19 pandemic, the findings suggest that patients with URTI-like symptoms were properly managed, and the rate of antibiotic usage remained reasonable.
Fulltext Transcriptome analysis of Rafflesia cantleyi flower stages reveals insights into the regulation of senescence

Mohd-Elias NA, Rosli K, Alias H, Juhari MA, Abu-Bakar MF, Md-Isa N, et al.

Sci Rep, 2021 Dec 08;11(1):23661.
PMID: 34880337 DOI: 10.1038/s41598-021-03028-x

Rafflesia is a unique plant species existing as a single flower and produces the largest flower in the world. While Rafflesia buds take up to 21 months to develop, its flowers bloom and wither within about a week. In this study, transcriptome analysis was carried out to shed light on the molecular mechanism of senescence in Rafflesia. A total of 53.3 million high quality reads were obtained from two Rafflesia cantleyi flower developmental stages and assembled to generate 64,152 unigenes. Analysis of this dataset showed that 5,166 unigenes were differentially expressed, in which 1,073 unigenes were identified as genes involved in flower senescence. Results revealed that as the flowers progress to senescence, more genes related to flower senescence were significantly over-represented compared to those related to plant growth and development. Senescence of the R. cantleyi flower activates senescence-associated genes in the transcription activity (members of the transcription factor families MYB, bHLH, NAC, and WRKY), nutrient remobilization (autophagy-related protein and transporter genes), and redox regulation (CATALASE). Most of the senescence-related genes were found to be differentially regulated, perhaps for the fine-tuning of various responses in the senescing R. cantleyi flower. Additionally, pathway analysis showed the activation of genes such as ETHYLENE RECEPTOR, ETHYLENE-INSENSITIVE 2, ETHYLENE-INSENSITIVE 3, and ETHYLENE-RESPONSIVE TRANSCRIPTION FACTOR, indicating the possible involvement of the ethylene hormone response pathway in the regulation of R. cantleyi senescence. Our results provide a model of the molecular mechanism underlying R. cantleyi flower senescence, and contribute essential information towards further understanding the biology of the Rafflesiaceae family.
Fulltext The Glaciozyma antarctica genome reveals an array of systems that provide sustained responses towards temperature variations in a persistently cold habitat

Firdaus-Raih M, Hashim NHF, Bharudin I, Abu Bakar MF, Huang KK, Alias H, et al.

PLoS One, 2018;13(1):e0189947.
PMID: 29385175 DOI: 10.1371/journal.pone.0189947

Extremely low temperatures present various challenges to life that include ice formation and effects on metabolic capacity. Psyhcrophilic microorganisms typically have an array of mechanisms to enable survival in cold temperatures. In this study, we sequenced and analysed the genome of a psychrophilic yeast isolated in the Antarctic region, Glaciozyma antarctica. The genome annotation identified 7857 protein coding sequences. From the genome sequence analysis we were able to identify genes that encoded for proteins known to be associated with cold survival, in addition to annotating genes that are unique to G. antarctica. For genes that are known to be involved in cold adaptation such as anti-freeze proteins (AFPs), our gene expression analysis revealed that they were differentially transcribed over time and in response to different temperatures. This indicated the presence of an array of adaptation systems that can respond to a changing but persistent cold environment. We were also able to validate the activity of all the AFPs annotated where the recombinant AFPs demonstrated anti-freeze capacity. This work is an important foundation for further collective exploration into psychrophilic microbiology where among other potential, the genes unique to this species may represent a pool of novel mechanisms for cold survival.

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