Presenter’s ID: OB02 Category: Non-Communicable Diseases TRADITIONAL CHINESE MEDICINE (TCM) BODY CONSTITUTION, FASTING BLOOD GLUCOSE AND WAIST CIRCUMFERENCE WITH CAPN10 (rs2975760) AND HNF1a (rs735396) Shu Rou Tan, Lai Kuan Teh Department of Allied Health Sciences, Universiti Tunku Abdul Rahman, Kampar, Perak. *Corresponding author email address: [email protected], [email protected] Metabolic syndrome (MetS) is a condition characterized by elevated blood glucose levels and increased waist circumferences, leading to type 2 diabetes mellitus (T2DM) and abdominal obesity. Traditional Chinese Medicine (TCM) categorizes individuals into different body constitutions, offering insights into disease prevention and diagnosis. Genetic variations such as single nucleotide polymorphisms (SNPs) in Calpain-10 (CAPN10) and Hepatocyte Nuclear Factor 1 Alpha (HNF1A) genes have been implicated in T2DM pathogenesis. This study contributes to a deeper understanding of the complex interplay between genetic variants, metabolic health and TCM body constitutions. A total of 80 subjects were recruited and demographic factors (gender, age and ethnicity) were considered. Interview-based TCM body constitution assessment was conducted, venous blood samples were collected for DNA analysis and fasting blood glucose (FBG) levels and waist circumferences were measured. IBM SPSS Statistical 26.0 was used for data analysis. This study found 1.25% and 24.20% prevalence of diabetes and abdominal obesity, respectively. Minor allelic frequency (MAF) for rs2975760 and rs735396 were greater than 0.05, indicating their significance in the study population. A weak positive correlation was found between FBG levels and waist circumferences, reflecting the impact of abdominal obesity on T2DM. Gender-based differences in FBG levels and waist circumferences were observed, possibly due to hormonal influences. FBG levels and waist circumferences differed significantly across five age groups, while no significant differences in FBG levels among ethnicities. There was no significant association between TCM body constitutions and genetic variations, suggesting the challenge of assessing the dynamic nature of body constitutions. Genetic variation like rs2975760 showed no significant differences in relation to FBG levels and waist circumference. The mutant CC allele of rs735396 was associated with significantly lower FBG levels but not for waist circumference. Further research involving larger and more diverse samples is recommended to fully understand the interactions between these variables. Keywords: Metabolic Risk Factors, Traditional Chinese Medicine Body Constitution, Genetic Variations 50
Presenter’s ID: OB03 Category: Non-Communicable Diseases A STUDY TO INVESTIGATE GLYCOLYSIS AND OXIDATIVE PHOSPHORYLATION DYNAMICS IN HEPG2 CELL-DERIVED 3D TUMOURSPHERES Li Qing Keoh , Sakunie Sawai , Thamil Selvee Ramasamy Department of Biomedical Science, Faculty of Medicine, Universiti Malaya, Wilayah Persekutuan Kuala Lumpur, Malaysia. Stem Cell Biology Laboratory, Department of Molecular Medicine, Faculty of Medicine, Universiti Malaya, Wilayah Persekutuan Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected] Hepatocellular carcinoma (HCC) is the most common primary liver cancer and the third leading cause of cancer death worldwide. Metabolic reprogramming is an emerging hallmark whereby cancer cells can alter their metabolism to adapt to various stress stimuli, thus conferring survival advantages. However, the metabolic switch between oxidative phosphorylation (OXPHOS) and glycolysis in HCC progression remains unclear. Therefore, in this study, we investigate the metabolic switch in HepG2 tumoursphere model. Firstly, a tumoursphere model was optimised using 4 different seeding densities (5000, 10000, 15000 and 30000 cells/well) of HepG2 cells in ultra-low attachment (ULA) condition over a 7-day culture course to generate viable and proliferative spheroids. The number of tumourspheres generated corresponded to the seeding density. HepG2 cells seeded at 15000 and 30000 cells/well formed tumourspheres of optimum size ( 150-200 µm), whereas the other 2 densities produced spheroids smaller than 150 µm. On day 7 of the culture, tumourspheres of lower seeding densities (5000-15000 cells/well) were viable with no dead cells detected. Collectively, a viable tumoursphere model was established using the seeding density of 15000 cells/well, concomitant with increasing relative percentage viability throughout the 7-day culture. Gene expression analysis of glycolytic genes (GLUT1, HK2, PFKFB3, PKM2) and OXPHOS genes (PGC1α, NRF1, TFAM, PDK1) was performed at selected timepoints (day 0, 1, 4 and 7) to examine the expression of selected metabolic genes in HepG2 tumourspheres. HK2 expression demonstrated a significant increase during tumoursphere progression, whereas the expression of GLUT1, PFKFB3, PKM2 and all OXPHOS genes were relatively comparable throughout the culture period. The current findings reveal that a 7-day HepG2 tumoursphere model established using the seeding density of 15000 cells/well generates a viable and proliferative 3D culture model, whereby HepG2 tumourspheres exhibit a propensity for glycolysis to adapt and proliferate in a new environment. Keywords: 3D culture, cancer, metabolic reprogramming 51 1 2 2* 1 2
Presenter’s ID: OB04 Category: Non-Communicable Diseases INVESTIGATING THE INTERSECTION: DEMOGRAPHIC PROFILES AND MICRORNA EXPRESSION IN BREAST CANCER Norlizawati Abdul Hamed , Tan Geok Chin , Cheah Yoke Kqueen Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected] Breast cancer (BC) is the predominant and most fatal female neoplasm in Malaysia, and is characterized by diverse molecular subtypes. MicroRNAs (miRNAs) regulate BC pathogenesis; however, their interaction with demographic factors in Malaysian populations remains underexplored. This study aimed to investigate the relationship between demographic profiles and miRNA expression patterns in BC patients from the Hospital Canselor Tuanku Mukhriz UKM (HCTM). Demographic data, including age, ethnicity, state, and clinical characteristics, were collected from 2023 patient records. Additionally, a systematic review was conducted to identify miRNAs associated with BC. Preliminary analysis revealed diverse demographic profiles among Malaysian BC patients, with 22.4% below 50 years old and 77.6% aged 50 and above, with a median age of 62. The ethnicity distribution comprised 57.3% Malay, 33.2% Chinese, 5.4% Indian, and 4.1% classified as 'Others'. BC subtypes included luminal A accounted for 29.5%, followed by luminal B (19.9%), triple-negative (14.1%), and Her-2 enriched (8.7%). In Malaysia's West Coast, the Central Region represented 36.4%, the Southern Region (22.9%), the Northern Region (20.0%), the East Coast (15.9%), and East Malaysia had 1.2%. Specific miRNAs identified in this systematic review, such as miR-27a, miR-944, and miR-125b, exhibited expression patterns predominantly observed in Chinese patients, reflecting findings from studies in China. Diverse demographic profiles emphasize the importance of personalized approaches in diagnosis and treatment. These findings indicate the need for further exploration of the effects of demographic factors and molecular subtypes on BC progression. The identification of specific miRNAs with expression patterns observed in Chinese patients, suggests potential ethnicityspecific differences in BC pathogenesis that require further investigation. In conclusion, this study highlights the complex relationship between demographic profiles and miRNA expression in Malaysian BC, emphasizing the significance of personalized treatment approaches by understanding the differences associated with ethnicity and molecular subtypes. Keywords: Breast cancer, demographic profiles, microRNA 52 1 2 1* 1 2
Presenter’s ID: OB05 Category: Non-Communicable Diseases DRP1 EXPRESSION PATTERNS IN BENIGN AND MALIGNANT BREAST LESIONS: A MALAYSIAN STUDY Leong Wen Kang , Phyu Synn Oo , Lwin Mie Aye , Noor Hasni Shamsudin , Nuramira Syazreen Suhaimi , Purushotham Krishnappa , Thin Thin Win School of Medicine, IMU University, Kuala Lumpur, Malaysia. Department of Pathology, School of Medicine, IMU University, Kuala Lumpur, Malaysia. Public Health and Community Medicine Department, School of Medicine, IMU University, Kuala Lumpur, Malaysia. Pathology Department, Hospital Tuanku Ja’afar, Seremban, Negeri Sembilan, Malaysia. School of Postgraduate Studies, IMU University, Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected] Breast cancer is the most prevalent cancer among Malaysian women. Approximately 70- 75% of invasive carcinoma shows oestrogen receptor (ER) positivity. Dynamin-related Protein 1 (Drp1) plays a crucial role in mitochondrial dynamics and has been implicated in various cancers, including breast cancer. It has been shown that oestrogen and ER enhances mitochondrial fragmentation and cell proliferation via Drp1 post-translational modification. Recent studies suggest Drp1 as a potential prognostic factor in breast cancer. However, there are no studies investigating the prevalence of Drp1 expression in ERpositive breast cancer in Malaysia. The objective of our study is to demonstrate the expression status of Drp1 in ER-positive breast cancer and benign breast lesions in the Malaysian population. A total of 97 breast lesion samples and patients’ data from pathology reports were collected randomly from the Pathology Department, Hospital Tuanku Ja’afar, after NMRR approval. The protein expressions of ER-α and Drp1 were assessed using immunohistochemistry (IHC). Patients’ data were analysed by SPSS. The age range of patients is between 18 – 76 years, and the mean age at diagnosis was 47±16.570 years. Among 97 patients, 75.3% (n=73) are Malay, 11.3% (n=11) are Chinese, and 13.4% (n=13) are Indian. Benign lesions (n=48) include fibroadenoma (n=23), fibrocystic disease (n=8), normal breast tissue (n=3), and other benign lesions (n=6). Malignant lesions include invasive ductal carcinoma (no special type) (n=38, 77.6%), invasive lobular carcinoma (n=6, 12.2%), ductal carcinoma in situ (n=2, 4.1%), mucinous carcinoma (n=2, 4.1%) and solid papillary carcinoma (n=1, 2.0%). Among women under 40 years old (n=34), 17.6% (n=6) present with malignant tumours while 68.3% (n=43) present with malignant tumours among women over 40 years old (n=63). Preliminary findings indicate that high immunoreactivity of Drp1 was correlated with ER-positive invasive breast carcinoma, while normal mammary glands show low immunoreactivity. Keywords: Drp1, breast cancer, prognosis 1 2 3 4 5 1 2* 3 4 2,5 2 2 53
Presenter’s ID: OB06 Category: Non-Communicable Diseases CYTOTOXIC EFFECT OF MALABARICONE C AGAINST HUMAN ORAL CANCER CELLS Nor Hafiza Rosli , Syarifah Nur Syed Abdul Rahman , Muhamad Aqmal Othman , Siti Amalina Inche Zainal Abidin , Nur'ain Salehen Department of Biomedical Science, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, Malaysia. Department of Oral and Craniofacial Sciences, Faculty of Dentistry, Universiti Malaya, Kuala Lumpur, Malaysia. Department of Chemistry, Faculty of Science, Universiti Malaya, Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected] Oral cancer is a non-communicable disease that has emerged in recent trends with significant treatment challenges, necessitating the exploration of new therapeutic strategies. Amidst the current trend going towards natural compounds as alternatives to conventional therapy, there is an opportunity to uncover the potential use of our native natural resources. Malabaricone C, a constituent of Myristica cinnamomea King was assessed for its potential cytotoxic activity against HSC-2 and HSC-3, human oral squamous carcinoma cell lines. The cytotoxic effect of malabaricone C was evaluated using an MTT assay, revealing a significant dose-dependent decrease in cell viability. Malabaricone C exhibited a lower IC50 value of 8.55 ± 2.32 µg/ml against HSC-2 compared to HSC-3 after 72 hours of incubation. Furthermore, safety evaluation in HGF, normal human gingival fibroblasts revealed weak cytotoxic effects of malabaricone C with IC50 value exceeding 100 μg/ml. Cellular and nuclear morphology changes of HSC-2 treated with malabaricone C were further observed via phase contrast microscopy and Hoechst33342/PI fluorescent staining. Morphology changes indicative of apoptosis, including plasma membrane blebbing, formation of apoptotic bodies, cell shrinkage, echinoid spikes, and nuclear fragmentation, were noticed in HSC-2 cells at 24 hours post-treatment. Malabaricone C dose-dependently induced the intrinsic (mitochondrial) apoptotic pathway in the HSC-2 cell line, involving loss of mitochondrial membrane potential (MMP), caspase 3 activation, and the presence of TUNEL-positive cell populations. Furthermore, the investigation into the cytotoxic activity of malabaricone C is currently ongoing, involving the analysis of its effects on cell cycle status to determine the specific phase in which the compound induces arrest as part of this project. These findings are anticipated to illuminate the promising potential of malabaricone C as an alternative medicine against cancer, providing valuable insight into its mechanisms of action at the cellular levels and thus laying a foundation for future research. Keywords: Malabaricone C, cytotoxic effect, oral cancer 54 1 2 3 1 2 3 2 1*
Presenter’s ID: OB07 Category: Non-Communicable Diseases THE EFFECT OF LITHIUM TOWARDS MOTOR SKILLS AND NEUROBEHAVIOURAL CHANGES IN ADULT MICE Bing-Cheng Goh , Mansour Azimzadeh , Pike-See Cheah , King-Hwa Ling Department of Biomedical Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. Department of Human Anatomy, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. *Corresponding author email address: [email protected] Down syndrome (DS) is one of the most common developmental disorders caused by chromosome 21 trisomy. Insulin resistance is reported to affect the cognitive functions in DS due to the dysregulated insulin signalling-related pathways. Lithium can act on the insulin signalling pathways and modulate various homeostatic mechanisms that affect oxidative stress and mitochondrial function. The study aims to examine the effects of lithium treatment (16mg/kg/day) on the motor skills and neurobehavioral changes of 3- month-old postnatal C57BL/6 disomic mice. Three-month-old C57BL/6 mice were randomly assigned into control (n=10) and lithium-treated (n=11) groups. Via oral gavage, the control group was fed with saline, and the lithium-treated group was fed with lithium chloride (16mg/kg/day) for 30 days. Motor skills and neurobehavioural functions were evaluated through hanging wire, rotarod, open field test (OFT), novel object recognition test (NORT) and Morris water maze experiments. Data was collected and analysed using ANY-maze and GraphPad Prism. Lithium-treated mice did not differ significantly from the control group on hanging wire, rotarod and Morris water maze tests. However, the lithiumtreated mice demonstrated a significantly improved behavioural performance in OFT (p<0.05) and NORT (p=0.02). In disomic mice, lithium treatment exerted a significant effect in reducing anxiety-related behaviours, which is consistent with prior studies that reported lithium as a mood-stabilising drug. Excitingly, lithium also showed a positive effect on short-term recognition memory, suggesting lithium possesses neuroenhancement properties. The neuroprotective effect of lithium in modulating homeostatic mechanisms can be further investigated in a trisomic mouse model. In conclusion, administration of lithium chloride to adult disomic mice for 30 days exerts notable positive effects on antianxiety and short-term memory improvement. Keywords: Lithium, neurobehavioural outcome, insulin resistance 55 1 2 1 1 2 1*
Presenter’s ID: OB08 Category: Non-Communicable Diseases DEVELOPMENT OF HEPG2-DERIVED SPHEROID MODEL FOR STUDYING DRUG RESPONSE IN HEPATOCELLULAR CARCINOMA (HCC) Wee Ling Choe , Ain Zubaidah Ayob , Thamil Selvee Ramasamy Department of Biomedical Science, Universiti Malaya, Kuala Lumpur, Malaysia. Department of Molecular Medicine, Universiti Malaya, Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected] Hepatocellular carcinoma (HCC) is often diagnosed at an advanced stage of the disease. Systemic therapy is a common treatment option for patients with advanced HCC; however, therapeutic resistance remains a challenge. In the discovery of better therapeutics, in vitro drug screening offers a crucial step in identifying drug candidates with the potential for further investigation in animal testing and clinical trials, however, conventional two-dimensional (2D) cell culture systems have major limitations in exploring the true potential of drugs. Therefore, this project aimed to develop a more representative three-dimensional (3D) culture model as a cell-based drug screening system to assess tumour response toward HCC-specific drugs. First, HepG2 cells were cultured at different seeding densities for up to 7 days in ultra-low attachment plates to form spheroids, and their morphology, viability and proliferation were characterized. The IC50 value of doxorubicin was determined using a monolayer culture, and the spheroids were treated with these concentrations to assess the drug response through viability and proliferation assays. We found that a seeding density of 500 cells/well was optimal to generate viable spheroids of 317.4 ± 7.4 µm diameter, with a progressive growth state. Day 4 was determined as the optimal time point for drug treatment. HepG2 spheroids exhibited 72% viability when treated with 12.5µM doxorubicin for 24h, which was 22% higher than that of the 2D culture, indicating lower efficacy of doxorubicin in the 3D culture at 24h. These results demonstrate that the developed HepG2 spheroid model requires a longer time to achieve a 50% reduction in the cell population. This may be due to drug resistance within the spheroids, suggesting that it is a more representative drug screening model that better mimics in vivo tumour responses. Keywords: 3D culture, drug response, hepatocellular carcinoma 56 1 2 1 2 2*
Presenter’s ID: OB09 Category: Non-Communicable Diseases THE CYTOTOXIC AND ANTI-INVASIVE EFFECTS OF AURANOFIN AND LUTEOLIN ON COLORECTAL CANCER CELLS Chang Joe Yan, Nurul Akmaryanti Abdullah, Noraina Muhamad Zakuan, Nur Fariesha Md Hashim Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia. Colorectal cancer (CRC) is the third most common and second most deadliest cancer worldwide. Anticancer drugs have emerged as crucial treatment strategies for CRC. Auranofin is a gold complex that primarily targets thioredoxin reductase (TrxR). However, its inhibition results in the activation of nuclear factor erythroid 2-related factor 2 (Nrf2), which upregulates various antioxidant enzymes to protect cells against oxidative stress. This compensatory mechanism could result in the chemoresistance of tumour cells against auranofin. Luteolin, a dietary flavonoids derived from vegetables and fruits, has been shown as a Nrf2 inhibitor. The dual pharmacological use of auranofin and luteolin is a novel strategy to inhibit antioxidant systems in cancer cells by increasing oxidative stress, ultimately resulting in cancer cell death. This study aimed to investigate the cytotoxic and anti-invasive effects of auranofin and luteolin in HCT116 CRC cells. HCT116 cells were seeded in a 96-well plate and treated with varying concentrations of auranofin and luteolin for 24 h followed by incubation with 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT). The IC20 and IC50 values were determined followed by the calculation of the combination index (CI) values using CompuSyn. Transwell invasion assay was conducted to assess the invasiveness of CRC cells. The percentage of HCT116 cells was significantly increased when treated with a combination of auranofin and luteolin (p=0.003) compared to single-agent treatment (p<0.001). The combination of 2µM auranofin and 5µM luteolin resulted in the lowest CI value (0.47), indicating the highest synergistic killing of cancer cells. The percentage of invading cells was significantly reduced in the combination treatment group (p<0.001). Dual combination treatment with auranofin and luteolin resulted in synergistic cell killing and a reduction in HCT116 cell invasiveness. Synergism of auranofin and luteolin can significantly exert cytotoxic and antiinvasive properties in HCT116 CRC cells compared with a single agent. Keywords: Auranofin, colorectal cancer, luteolin 57
Presenter’s ID: OB10 Category: Non-Communicable Diseases CYTOTOXIC EFFECT OF MALABARICONE B ON HUMAN ORAL CANCER CELLS Dhamirah Irdina Mohammad Azzuddin , Syarifah Nur Syed Abdul Rahman , Muhamad Aqmal Othman , Siti Amalina Inche Zainal Abidin , Nur’ain Salehen Department of Biomedical Science, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, Malaysia. Department of Oral and Craniofacial Sciences, Faculty of Dentistry, Universiti Malaya, Kuala Lumpur, Malaysia. Department of Chemistry, Faculty of Science, Universiti Malaya, Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected] Oral cancer is an emerging non communicable disease that causes significant burden in the healthcare system globally due to the high number of incidence and mortality. Malabaricone B, a phytochemical which is a major constituent of acylphenols in Myristica cinnamomea King, has demonstrated promising anticancer properties in various cancer types. However, the cytotoxic effect of malabaricone B on human oral cancer cells have not been elucidated yet. This study aims to evaluate the cytotoxic effect of malabaricone B against human oral cancer cells. In vitro cytotoxicity of malabaricone B on HSC-3 and HSC-2 human oral squamous carcinoma cells and a normal hGF human gingival fibroblast cells was evaluated using MTT assay. The result showed significant cytotoxicity of malabaricone B against both HSC-2 and HSC-3 cells with the IC50 values of 9.5 ± 2.32 µg/mL and 17.5 ± 2.03 µg/mL respectively. Following that, cellular and nuclear morphological alterations in HSC-2 cells were observed using phase contrast microscopy and Hoechst 33342/PI double staining. The results revealed significant cellular and nuclear morphological changes, supporting the presence of apoptosis. To validate these findings, biochemical changes related to apoptosis were assessed. Specifically, the evaluation focused on measuring the depolarization of mitochondrial membrane potential (MMP) and caspase-3 activity through MMP assay and caspase-3 assay to validate the morphological findings. The results revealed a dose-dependent increase in MMP depolarization, accompanied by the activation of caspase-3. In conclusion, Malabaricone B shows selective cytotoxicity towards oral cancer cells, with a pronounced effect on HSC-2 cells. It induces apoptosis in oral cancer cells through the intrinsic pathway, evidenced by MMP depolarization and caspase-3 activation. Ongoing investigations focusing on DNA fragmentation and cell cycle arrest to further validate these findings. These results may further elucidate the potential use of malabaricone B as a novel chemotherapeutic agent for combating human oral cancer. Keywords: Cytotoxicity, malabaricone B, oral cancer 58 1 2 3 1 2 3 2 1*
Presenter’s ID: OB11 Category: Non-Communicable Diseases THE ANTI-INVASIVE EFFECTS OF 1-METHYLPROPYL 2-IMIDAZOLYL DISULFIDE (PX-12) ON HYPOXIA-INDUCED COLORECTAL CANCER CELLS AND ITS UNDERLYING MECHANISM OF ACTION Chong Qiao Yi, Nurul Akmaryanti Abdullaha, Noraina Muhamad Zakuana, Nur Fariesha Md. Hashim Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia. According to the World Health Organization, colorectal cancer (CRC) is the second most common cancer that leads to fatality globally and holds the fourth-highest mortality rate among the other cancers in Malaysia. Cancer malignancy is characterised by metastasis, which is driven by invadopodia formation and epithelial-mesenchymal transition (EMT). 1-methyl propyl 2-imidazolyl disulfide (PX-12) has been shown to have anti-cancer effects by targeting the thioredoxin (Trx) system. However, the anti-invasive effects and its underlying mechanism against CRC under hypoxia conditions are limited. This study aims to investigate the anti-invasive effect of PX-12 on invadopodia formation and its effects on EMT-related proteins. HCT116 cells were seeded into 96-well plate prior to 3-(4, 5- dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay to determine PX-12’s inhibitory concentrations. Cells in the hypoxia group were treated with a hypoxia mimetic agent, 1mM (Dimethyloxalylglycine) DMOG, to mimic hypoxia condition. In gelatin degradation assay, cells were treated with various concentrations of PX-12 (0μM, 2.5μM, and 5μM) for 24 hours under normoxia and hypoxia conditions followed by seeding on the coverslip coated with Oregon Green 488-gelatin. Western blot was used to determine the expression levels of hypoxia-inducible factor 1-alpha (HIF1- α), Trx and EMT-related proteins. PX-12 significantly reduced the cell viability of HCT116 upon 24- hour and 48-hour incubation with the IC20 values ranging from 1.5µM to 4µM and IC50 values ranging from 11µM to 14µM in normoxic and hypoxic conditions. The gelatin degradation assays showed that PX-12 at 2.5uM and 5uM under hypoxic conditions significantly reduced the number of cells with invadopodia. Western blot analysis demonstrated a significant increase in HIF-1α expression in HCT116 cells treated with 1mM DMOG. The substantial reduction in invadopodia formation in HCT116 cells under hypoxia conditions demonstrated that PX-12 has anti-invasive effects. Keywords: Anti-invasive, colorectal cancer, PX-12 59
Presenter’s ID: OC01 Category: Others IN VITRO AND IN SILICO ANALYSIS OF LAWSONE AND ITS ANALOGUE (2- BROMO-1,4- NAPHTHOQUINONE) AS POTENTIAL THERAPEUTIC AGENTS IN WOUND HEALING Muhammad Faiz bin Abdul Samad , Seri Narti Edayu Sarchio, Siti Farah Md Tohid Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor. *Corresponding author email address: [email protected] Several studies have been conducted to develop wound healing agents from natural resources. Lawsone (2-hydroxy-1,4-naphthoquinone), a natural compound found in Lawsonia inermis or henna, has demonstrated promising wound healing properties in earlier studies by interfering with the healing pathways through interactions with a protein called aryl hydrocarbon receptor (AhR). Besides, one of its synthetic analogue (2-bromo1,4-naphthoquinone), has been previously shown to possess anticancer and wound healing potentials. Treating wounds can be challenging as current treatments have limitations in terms of the efficacy, potency and safety. Therefore, exploration of novel agents like lawsone and its analogue poses a necessity to develop better treatment options. Investigating both compounds for potential wound healing properties may provide valuable insights into alternative, effective and safer therapeutic strategies for treating wounds. This study aims to evaluate the potential of lawsone and 2-bromo-1,4- naphthoquinone as therapeutic agents in wound healing via cell migration assay as well as to assess the toxicity of both compounds via MTT assay in vitro. Besides, molecular interactions with AhR between lawsone and 2-bromo-1,4-naphthoquinone in wound healing process will also be evaluated by performing molecular docking in silico analysis. It is predicted that lawsone and 2-bromo-1,4-naphthoquinone may promote the migration of 3T3 cells in wound healing and exhibit safe toxicological profile for application. It is also hypothesized that lawsone and 2-bromo-1,4-naphthoquinone may exhibit strong binding affinity with AhR. By using 3T3 fibroblast cell line for in vitro studies, MTT assay was carried out to determine the cytotoxicity and safe doses of lawsone and 2-bromo-1,4- naphthoquinone, followed by scratch assay to assess the effects of both compounds on cell migration for wound closure. The in vitro results will be analyzed by using one-way ANOVA, with the Dunnett test as a post-hoc test which is significant at p<0.05. In silico molecular interaction analysis between lawsone or 2-bromo-1,4- naphthoquinone with AhR was utilized using molecular docking studies via AutoDock Vina. Their interactions were analyzed by Pymol software, ProteinPlus and PLIP web servers. The in vitro analysis of this study is currently ongoing. Lawsone and 2-bromo-1,4-naphthoquinone are expected to exhibit enhanced wound healing properties and good cell migrations with safe toxicity profiles. In in silico studies, both compounds demonstrated strong binding affinities to AhR at -7.6 kcal/mol for lawsone and -8.2 kcal/mol for 2-bromo-1,4-naphthoquinone through interactions with the amino acid residues Thr289, His291, Phe295, Ser365 and Gln383. Both results from ProteinPlus and PLIP web server showed that hydrophobic interactions occurred between lawsone and AhR residue Phe295D. Hydrogen bondings were also detected with Gly321D and Ser365D. On the other hand, one hydrophobic interaction was identified between 2-bromo-1,4-naphthoquinone and AhR at the residue Val381D and two hydrogen bondings were recognized with His291D and Gln383D by the PLIP web server. Structure activity relationship profile between lawsone and 2-bromo-1,4- naphthoquinone in enhancing wound healing activities will be established based on in vitro and in silico results. Keywords: Lawsone, 2-bromo-1,4-naphthoquinone, wound healing, scratch assay, AhR, molecular docking 60 *
Presenter’s ID: OC02 Category: Others CHARACTERIZATION OF PERIOSTIN 2 (OSF-2) ON THE ROSETTING PHENOMENON BY MALARIA PARASITES Zhi-Ying Phong , Wenn-Chyau Lee Department of Parasitology, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, Malaysia. Department of Biomedical Science, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, Malaysia. A*STAR Infectious Diseases Labs, Agency for Science, Technology and Research (A*STAR), Singapore. *Corresponding author email address: [email protected] Malaria is a potentially fatal infectious disease that is prevalent in many tropical countries. The pathogenesis of malaria attributes to the cytoadherence properties of the Plasmodiuminfected erythrocytes (IRBC) including rosetting and IRBC-endothelial cytoadherence. Rosetting phenomenon facilitates the IRBC to sequester within the vasculature to escape splenic filtration, hampers phagocytosis of the IRBC by the host phagocytes, and confers anti-malarial resistance to the late-stage-parasites. Previous study reported significant inhibition of rosette formation by antibodies against human periostin 2 (OSF-2). In this study, we aimed to investigate the role of OSF-2 in Plasmodium rosetting, characterize OSF-2 mediated rosetting ligands, and compare OSF-2 mediated rosetting rates under varying human serum concentrations. Several laboratory-adapted P. falciparum and P. knowlesi isolates were cultured and the late stages were purified using the Magnetic Activated Cell Sorter (MACS). The purified late-stages were mixed with the uninfected erythrocytes and OSF-2 for rosetting assay. OSF-2 of 200 ng/ml yielded consistent rosette stimulation on both species. In addition, we demonstrated the serum dependency of OSF-2 mediated rosetting. Furthermore, the rosette-stimulating effect of OSF-2 was completely abrogated after trypsinization of the IRBCs, suggesting a role played by PfEMP1 in the OSF-2 mediated rosetting by P. falciparum. For P. knowlesi, the rosetting ligand involved was highly sensitive to trypsin. Our work characterized the OSF-2 mediated rosetting by both P. knowlesi and P. falciparum. Keywords: Plasmodium, OSF-2 mediated rosetting, ligand 61 1 2 3 1,2 1,3*
Presenter’s ID: OC03 Category: Others CHARACTERISATION OF Staphylococcus aureus FROM THE NASOPHARYNX OF HEALTHY CHILDREN ATTENDING NURSERIES IN KLANG VALLEY Chew Man Fei, Tony Khoo Cheng How , Abeer Saeed Y Alsomali , Norfazlina Mohamad, Maitasha Alia Meor Yahaya, Nurshahira Binti Sulaiman , Mohd Nasir Bin Mohd Desa Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. Staphylococci are Gram-positive bacteria that possess a round shape and tend to aggregate in clumps. Staphylococci are part of the normal flora that colonise the human nasal passages. Staphylococcus aureus (S. aureus) is an opportunistic pathogen capable of causing a variety of clinical manifestations when the host's immune system is compromised. Overuse of antibiotics in humans causes the development of drug-resistant pathogens. Methicillin-resistant Staphylococcus aureus (MRSA) has been found to escape from the hospital settings and disseminate into community. This study aims to characterise the phenotypic and molecular aspects of S. aureus isolates obtained from the nasopharynx of healthy children. A cohort of 124 healthy children of 5 years old and less attending nurseries in Klang Valley were previously sampled in a surveillance study involving nasopharyngeal swap. This current study further screened the samples for S. aureus using standard biochemical tests, followed by antimicrobial susceptibility profiling against erythromycin, vancomycin cefoxitin ciprofloxacin and gentamicin and mecA gene detection by PCR for identifying MRSA. Selected MRSA isolates will be subjected to multilocus sequence typing (MLST). 46 out of 124 individuals were found to carry S. aureus (37.10%). Among these 46 isolates, antimicrobial susceptibility analysis revealed that 15.22% (7/46) and 8.70% (4/46) were resistant and intermediate susceptibility to erythromycin respectively. Additionally, 2 isolates (4.35%) demonstrated resistance to ciprofloxacin and 1 isolate (2.17%) showed resistance to cefoxitin. The presence of the mecA gene was detected in 3 isolates (6.52%) which indicate the presence of MRSA. The occurrence rate of S. aureus at 37% exceeds the range reported by Mohamed et al. (2016) which is 18% - 30%. The detection of MRSA in the cohort population underscores the persistent challenge posed by resistant strains, which can complicate clinical management and escalate healthcare costs. The presence of the mecA gene highlights the need for vigilant surveillance and infection control measures to prevent the spread of MRSA among community. Despite the relatively low rate of MRSA, the risk of its spread remains significant, especially in the context of poor hygiene practices. Nurturing awareness about the risks of S. aureus infections and advocating for hygienic lifestyles among both children and parents is crucial. Keywords: Staphylococcus aureus, characterisation, antibiotic 62
Presenter’s ID: OC04 Category: Others INVESTIGATION OF MICROBIOTA COMMUNITY IN THE ADULT STAGES OF Aedes aegypti MOSQUITOES Zi Xian Voon, Min Jie Chung, Lai Yee Ho Department of Allied Health Sciences, Faculty of Science, Universiti Tunku Abdul Rahman, Kampar, Perak, Malaysia. *Corresponding author email address: [email protected], [email protected], [email protected] Chikungunya and dengue fever are two of the most typical mosquito-borne endemic diseases present nowadays, especially in Malaysia. In the first month of year 2024, the number of dengue cases had increased to 18,247 cases in Malaysia. Even though these diseases existed for decades, an effective control method that can minimize their transmissions by their vectors, Aedes aegypti and Aedes albopictus is not yet well established. This research aims to investigate the internal microbiota present in the Aedes aegypti. It is a preliminary study to investigate the possibility of disrupting their vectorial capacity through the modification of their internal microbiota. Lab strain Ae. aegypti eggs were purchased from the Vector Control Research Unit (VCRU), Universiti Sains Malaysia (USM), Penang. Following a series of morphological and biochemical identifications, a total of three isolates (M1-M3) were obtained from the male Ae. aegypti, and three isolates (F1-F3) were obtained from the female Ae. aegypti. Subsequently, DNA extraction using a commercial kit for obtained isolates was performed, followed by 16S polymerase chain reaction (PCR), then purified and sent for DNA sequencing. These isolates obtained are all catalase positive which also shows a variety of Gram stain characteristics and the different ability to form spores. The unique colony morphology of isolates and their biochemistry characteristics also further visualize the identity of isolates. The results showed that the isolates obtained were Bacillus spp., Enterobacter spp., Pantoea spp., and Phytobacter spp. Further study of these bacteria should be done to understand their relationship with the mosquitoes and how they would influence the mosquitoes’ capacity as vector. Keywords: Internal microbiota, Aedes aegypti, Bacteria identification 63
Presenter’s ID: OC05 Category: Others CHARACTERIZATION OF COAGULASE-NEGATIVE STAPHYLOCOCCUS (CONS) FROM THE NASOPHARYNX OF HEALTHY CHILDREN ATTENDING NURSERIES IN KLANG VALLEY Tony Khoo Cheng How, Chew Man Fei, Abeer Saeed Alsomali, Norfazlina Mohamad, Maitasha Alia Meor Yahaya, Nurshahira Sulaiman, Mohd Nasir Mohd Desa Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, University Putra Malaysia, Selangor, Malaysia. Coagulase-negative Staphylococci (CoNS) are a group of bacteria that belongs to the genus Staphylococcus and lacks the ability to produce coagulase enzymes. Concurrently, a prominent issue has emerged concerning the role of drug-resistant CoNS, particularly methicillin-resistant CoNS (MRCoNS), as the reservoir for transmitting the antibioticresistant traits to other Staphylococcus spp. via conjugative plasmids. It is also believed that methicillin-resistant Staphylococcus aureus (MRSA), which is better suited for causing infections and diseases in humans, has served as the recipient and disseminator among human populations. Consequently, investigating the occurrence of MRCoNS in community settings has become the primary focus of this study. This research aims to identify and characterize a collection of secondary bacterial stock obtained through nasopharyngeal swab from healthy children for coagulase-negative Staphylococci and their antimicrobial susceptibility features. In a previous surveillance study, a cohort comprising 161 healthy children aged 5 years or younger, attending nurseries in the Klang Valley, previously underwent nasopharyngeal swab sampling. In this current study, the samples were further analysed for CoNS through standard biochemical tests. Antimicrobial susceptibility profiling was conducted using disc diffusion methods against erythromycin, vancomycin, cefoxitin, ciprofloxacin, gentamicin, and penicillin G. Additionally, the mecA gene was detected using PCR to identify and confirm MRCoNS. Selected MRCoNS isolates were further subjected to multilocus sequence typing (MLST) to determine their sequence type (ST). Among the 161 subjects, 54 of them (33.54%) were identified for CoNS. Out of 54 CoNS, 14 of them (25.93%) showed methicillin resistance (MRCoNS) based on the results of cefoxitin disc and all of these methicillin resistant strains tested positive for the mecA gene by PCR. Among the 14 MRCoNS, 5 samples were chosen and sent for sequencing at 1st BASE, Malaysia. Based on the analysis of the sequencing results, one strain (TPJ 14/02) is classified as ST130 while the remaining 4 samples have no exact match to either of the ST available in the databases. Since 2007, ST130, originating from Europe, has been uncommonly found in Asian countries, primarily isolated through skin swabs from patients with unidentified diseases. Conversely, additional analysis and studies should be conducted on the remaining 4 samples exhibiting unmatched ST, as there is a possibility of new ST emergence among MRCoNS. While the prevalence of MRCoNS among children is comparatively low, its impact should not always be underestimated. Parents play a crucial role in educating their children about the significance of maintaining a hygienic lifestyle, as there remains a risk of MRCoNS spreading within the child population. Failure to properly control the spread of MRCoNS could greatly diminish the effectiveness of current treatments against Staphylococcal infections. Keywords: Coagulase-negative Staphylococci (CoNS), antimicrobial susceptibility profiling, mecA gene, multilocus sequence typing (MLST), sequence type (ST) 64
Presenter’s ID: OC06 Category: Others PRODUCTION AND CHARACTERIZATION OF CHITOSAN FROM Mucor circineloides Nur Arisya Qamarina, Nur Syahmina School of Health Science, Health Campus, University Sains Malaysia, 16150 Kubang Kerian, Kelantan. *Corresponding author email address: [email protected] The aim of this study was to determine the optimization of the factors that affects the quality and quantity of fungi-based chitosan (Mucor circinelloides) which are incubation temperature and incubation period applied during alkaline deacetylation by measuring the dry weight and degree of deacetylation of chitosan. Mucor circinelloides was cultivated on Potato Dextrose agar (PDA) and nutrient broth before the extraction of mycelium of chitosan. The chitosan in dried powder form then was treated with alkaline, Sodium Hydroxide (NaOH) and heated for deacetylation purposes at different incubation period and incubation temperature which were 90°C, 98°C, 110°C and 121°C for 15, 20, 25 and 30 minutes. Alkaline insoluble material obtained then was treated with acetic acid to remove unwanted materials. The obtained chitosan pellet was characterized by degree of deacetylation through acid-base titration method and the result obtained was analysed by Kruskal-Wallis test from IBM SPSS Statistics 24. From the analysation of result, when incubation period increased, it only showed a slight increase in terms of dry weight and degree of deacetylation which attribute to no significant differences (p>0.05) of incubation period on both variables. Meanwhile, when incubation temperature increased, dry weight and degree of deacetylation became decreases, whereas the incubation temperature also did not show significant differences on dry weight of chitosan but exhibited significant differences on degree of deacetylation which may happen due to unwanted chain termination or degradation of chitin or chitosan chain. The findings contribute valuable insights into their potential use but also exhibits limitations like variability of the parameters like degree of deacetylation. Therefore, further research is warranted to optimize the extraction process like by using deep eutectic solvents. Overall, it lays the groundwork for future studies of chitosan production from fungi, emphasizing the importance of sustainable and eco-friendly approaches in their development. Keywords: Mucor circineloides, Chemical Extraction, Degree of Deacetylation 65
Presenter’s ID: OC07 Category: Others THE SENSITISATION PROFILE OF MALAYSIAN ATOPIC POPULATION TO THREE MAJOR HOUSE DUST MITE ALLERGENS AND ITS ASSOCIATION WITH ALLERGIC DISEASES AND GEOGRAPHIC LOCATION Ng Yong Yee , Abosede Temitope Olajide , Intan Hakimah Ismail , Tham Chau Ling Cell and Signalling Laboratory, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. Clinical Immunology Unit, Department of Paediatrics, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. *Corresponding author email address: [email protected] The predominant allergenic house dust mites (HDM) species Dermatophagoides pteronyssinus (Der p), Dermatophagoides farinae (Der f), and Blomia tropicalis (Blo t) are prevalent in Malaysia due to the favourable climate and urbanisation trends. HDM triggers allergic diseases such as asthma, rhinitis, and atopic dermatitis, contributing significantly to the healthcare and economic burden. However, limited information exists on the sensitisation of these major HDM across different regions in Malaysia. This crosssectional study reported the epidemiological status of three HDM allergens among atopic populations and their association with allergic diseases in two regions of Malaysia. HDM allergy data of 366 atopic patients, aged 7-60 from four hospitals in the Klang Valley and East Coast were collected through questionnaires whereas their sensitization profiles were obtained through skin prick test (SPT) with Der p, Der f, Blo t, positive (histamine) and negative (glycerin saline) controls. The results were recorded after 15 min, with a wheal diameter >3 mm considered positive. The chi-square test was used to compare frequencies, with significance set at p<0.05. Of 354 patients, 257 (70.2%) were females and 109 (29.8%) were males. Sensitization rates to Der p, Der f and Blo t were 55.65%, 55.93% and 51.41% respectively, with 40.4% of the patients sensitized to all three HDM allergens. HDM sensitization was positively correlated with allergic diseases (p=0.009) and geographical distribution (p=0.035). Among the atopic population in Malaysia, the prevalence of the three types of house dust mites was relatively similar, with Der f being slightly higher. Patients with allergic rhinoconjunctivitis exhibited greater sensitivity to Blo t (p=0.02). Additionally, HDM sensitization was higher among patients residing in the Klang Valley (71.89%) than among those residing in the East Coast (59.17%). This study has enhanced the knowledge on HDM allergy epidemiology in Malaysian urban areas, serving as preliminary findings for specific diagnostic tools for HDM allergens. Keywords: prevalence, house dust mite (HDM), allergic diseases 66 1 2 1 1 2 1*
Presenter’s ID: OC08 Category: Others EXPRESSION, PURIFICATION AND CRYSTALLIZATION OF LEPTOSPIRA PROTEIN Noraishah Hairul Nizam , Mohd Anuar Jonet Biomedicine Programme, School of Health Sciences, Health Campus, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan. Malaysia Genome and Vaccine Institute (MGVI), National Institute of Biotechnology Malaysia (NIBM), Jalan Bangi, 43000 Kajang, Selangor. *Corresponding author email address: [email protected] Immunoglobulin-like protein LigB, is a surface-exposed protein found in pathogenic Leptospira species, which are the causative agents of leptospirosis. Leptospirosis is a globally significant zoonotic disease. With the study of the potential structure of LigB, researchers were able to study the targets for vaccine development. The protein expression was started by streaking the protein onto LB agar to isolate a single colony, from which a starter culture was prepared. Subsequently, a subculture was generated, and the protein expression was induced using IPTG. After harvesting and lysing the protein, Immobilized Metal Ion Affinity Chromatography (IMAC) purification was carried out using an AKTA System with centrifuged (8,000 rpm, 10 mins, 4°C) supernatant treated with binding buffer, (25 mM Tris, 150 mM NaCl, 20 mM Imidazole and 20% glycerol). Then, followed by size exclusion chromatography for further purification. Buffer exchange with 25 mM Tris was performed, and crystal screens (Crystal Screen 1 and 2 Research Hampton) with 96 different precipitants were conducted to identify suitable conditions for proteins to grow crystals. The screening was monitored regularly (daily and then weekly after a week of screening) to observe the crystals progress. 7.8 ml of protein sample was obtained from the expression procedure, which was then used for the purification part. A single band on SDS-PAGE analysis after size exclusion chromatography was obtained suggesting a successful separation based on size exclusion principles. The crystals that formed from screening were needle crystals, whisker crystals and plate crystals (stack or multilayer crystals). The SDS-PAGE that was performed on the crystal screen indicates the crystal was LigB protein. A hanging drop and sitting drop plates were performed using 0.1 M HEPES pH 7.5, 20% w/v Polyethylene glycol 10,000 as the precipitant. These findings from the crystal screen indicate that the diffraction X-ray studies will be promptly conducted. Good crystals are characterized by their shiny surfaces and uniform shapes indicating a high level of purity and ordered growth. A good precipitate in crystallization is a solid that forms quickly throughout the solution, typically appearing cloudy and less defined compared to crystals. Keywords: Leptospira, LigB protein, Crystallization 67 1 2 1* 2
Presenter’s ID: OC09 Category: Others PREVALENCE OF MICROBIAL SPECIES IN AIR, SURFACES AND SKIN AMONG CHILDREN IN TASKA CAHAYA MATA, FAKULTI PERUBATAN DAN SAINS KESIHATAN AND TASKA CAHAYA MATA, HOSPITAL SULTAN ABDUL AZIZ SHAH Muhammad Firdaus Bin Abd Rahman , Elysha Nur Binti Ismail , Juliana Binti Jalaludin , Mohd Nasir Bin Mohd Desa , Reezal Bin Ishak , Zulkefley Bin Othman Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. Department of Environmental and Occupational Health, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. Universiti Kuala Lumpur Institute of Medical Science Technology, Kajang, Selangor, Malaysia. *Corresponding author email address: [email protected] Microbes are present in our surroundings including surface, air, and skin. The surroundings, including indoor environments, greatly influence human microbial exposure and colonization. Nevertheless, microbes are also found to be common in the human body which is known as normal flora. However, some strains of microbes can be pathogenic to humans, especially in children. This study aims to investigate the microbial composition of air, surfaces, and skin sources among children within Taska Cahaya Mata FPSK and Taska Cahaya Mata, HSAAS. The samples were collected from three parameters which were air, surfaces, and skin of the children ages 1 to 4 years old that attend the daycare center. 41 samples of surfaces and 17 samples of children’s skin were collected by swabbing method using cotton swabs while 10 samples of air were taken by using SAS Super 100 microbiological air sampler. All the samples taken were cultured on mannitol salt agar (MSA) and Macconkey agar respectively. The antimicrobial susceptibility of the isolates from the sample were tested by using Kirby-Bauer disc diffusion method. Further analysis for molecular confirmation was carried out by using the monoplex PCR method to amplify the mecA gene for detection of methicillin-resistant Staphylococcus aureus. Among the total of 68 samples that were collected from both daycare centers, 35.3% (24/68) were mannitol fermenters and 44.12% (30/68) were nonmannitol fermenters on mannitol salt agar. The bacterial culture on Macconkey agar showed 7.35% (5/68) were lactose fermenters and 14.71% (10/68) were non-lactose fermenters. Three samples were found to be resistant towards oxacillin and cefoxitin and show presence of mecA gene. These findings show that the environment of daycare centers can influence the colonization of the microbes on the skin of children that attend the daycare centers despite limited sample size and collection sample period. Keywords: microbial species, skin of children, environment of daycare center’s 68 1 2 3 1* 1 2 1 3 1
Presenter’s ID: OC10 Category: Others IN SILICO AND ANTIBACTERIAL EFFECTS OF ANDRICIN 01 AGAINST Staphylococcus aureus Jesther Chong See Mun , Anis Rageh Al-Maleki Department of Biomedical Science, Universiti Malaya, Kuala Lumpur, Malaysia. Department of Medical Microbiology, Universiti Malaya, Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected] Staphylococcus aureus are Gram-positive cocci in clusters that can cause various diseases with their virulence factors. The emergence of antibiotic resistant strains has threatened the benefits of antibiotics, which lead to poor treatment outcomes and contribute to the global burden. Therefore, antimicrobial peptides (AMPs) can be a feasible alternative treatment to combat antimicrobial resistant infections as their efficacies have been demonstrated in recent papers. Andricin 01 is a novel AMP that is found in the mucus of a Chinese giant salamander (Andrias davidianus). This research aims to investigate the antibacterial effect of Andricin 01 against S. aureus in silico and in vitro. In this study, the interactions between Andricin 01 and various virulence gene receptors of S. aureus were investigated using molecular docking. Among all receptors, Andricin 01 bound best with 1N67 with the lowest binding affinity of -8.1 kcal/mol. It was found that Andricin 01 bound to the 1N67 receptor at ASN-343, ASN-284, LEU-285 and GLY-313. S. aureus was identified via culture, catalase test, tube coagulase test, and 16S rRNA polymerase chain reaction (PCR). The identity of S. aureus was confirmed by their culture characteristics, biochemical test and molecular approaches. S. aureus exhibited small, round, golden or yellow colour colonies on blood nutrient agar. It was positive for both the catalase test and tube coagulase test. Gradient PCR with annealing temperatures between 50°C to 60°C found that the best annealing temperature for S. aureus was at 50.7°C. The minimum inhibitory concentration (MIC) of Andricin 01 against S. aureus following the Clinical and Laboratory Standards Institute (CLSI) broth microdilution was 62.5 µg/mL. Therefore, Andricin 01 showed good interaction stability with 1N67 and promising antibacterial effects on S. aureus. Thus, it can be considered as a drug candidate for antimicrobial infections. Keywords: Molecular docking, Staphylococcus aureus, Andricin 01 1 2 1 2* 69
Presenter’s ID: OC11 Category: Others EXPLORING THE MODULATORY EFFECTS OF LAWSONE AND ITS DERIVATIVE MENADIONE ON CELL MIGRATION IN WOUND HEALING: AN IN VITRO AND IN SILICO STUDIES Hariz Syahmi Hazandy, Seri Narti Edayu Sarchio, Siti Farah Md Tohid Department of Biomedical Science, Faculty of Medicine and Health Sciences, UPM, Selangor, Malaysia. *Corresponding author email address: [email protected] Wound healing is a complex biological process that results in the recovery of tissue integrity after an injury. Fibroblasts are cells that play a central role in all phases of wound closure. The search for an ideal treatment for wounds is still constantly evolving. Currently, there are many modern treatments available to treat wounds, however, they are still hindered by various problems of unfavourable side effects and lack of potency and efficacy. Hence, numerous studies have been performed to investigate the potential woundhealing properties of natural products obtained from plants as alternative medicine. For instance, henna which is derived from the leaves of Lawsonia inermis has been traditionally prescribed in Ayurvedic medicine as a topical treatment for various skin and hair conditions. Lawsone (2-hydroxy-1,4-naphthoquinone) is the natural pigment found in the henna leaves which is responsible for the red-orange dyeing properties of henna. Scientifically, lawsone possesses various pharmacological properties including antifungal, antibacterial, antiviral, anti-inflammatory, anticancer, antispasmodic, and wound healing. While menadione (2-methyl-1,4-naphthoquinone), is a derivative of lawsone which has been reported to possess anticancer, antiparasitic, antichagasic, antimalarial and antimicrobial potentials. This study aims to investigate the cell migratory effects of lawsone and its derivative menadione on 3T3 fibroblast cells as well as assess their cytotoxicity effects in vitro. Their mechanistic ability to interact with TGF-β1 in silico will also be investigated. For the in vitro study, the IC20 and IC50 of lawsone and menadione on 3T3 fibroblast cells will be determined through an MTT assay. The MTT assay results will then be employed to determine the treatment doses, which will subsequently be utilised to assess the effects of both compounds on the cell migration rate of 3T3 fibroblast cells through cell migration assay. The in vitro results will be analysed by using one-way ANOVA, with the Dunnet test as a post-hoc test which is significant at p<0.05. Subsequently, the in-silico studies which involve the application of AutoDock Vina software to predict the interactions between lawsone and menadione towards TGF-β1 protein were performed. The interactions of lawsone, menadione and TGF-β1 were analysed by Pymol software, PLIP and ProteinPlus webservers. For in silico studies, lawsone and menadione showed strong binding affinities to the TGF-β1 protein at -7.9 and -8.4 kcal/mol respectively. Results from the PLIP webserver illustrated several hydrophobic and hydrogen bonding interactions. The hydrophobic interactions between lawsone with five protein residues; Val219A, Tyr249A, Leu260A, Phe262A, and Leu278A, while the hydrophobic interactions between menadione were with six protein residues; Val219A, Lys232A, Tyr249A, Leu260A, Phe262A, and Leu278A. Additionally, for lawsone, there were three hydrogen bonding with Ala230A (3.19Å), Lys232A (3.21Å) and Asp351A (3.10Å). Meanwhile, menadione had two hydrogen bonding with Lys232A (3.25Å) and Asp351A (3.07Å). Although, in vitro studies are currently underway and will be reported in due course. It is expected that lawsone and menadione will have significant effects on cell migration in the cell scratch assay. Additionally, the IC20 and IC50 of both compounds are expected to show relatively low cytotoxicity effects on 3T3 fibroblast. Both the in silico and in vitro results will assist in constructing a structure-activity relationship of lawsone and menadione in accelerating wound healing processes. Keywords: Wound healing, Lawsone, Menadione, Molecular docking, Scratch assay 70 *
Presenter’s ID: OC12 Category: Others REVITALIZING PULP: BMP-7'S INFLUENCE ON WJMSC AND DHDP THROUGH HISTOLOGY AND VIABILITY Nur Athirah Ahmad Shuhaimi , Farinawati Yazid , Dalia Abdullah , Ng Sook Luan , Nurul Inaas Mahamad Apandi Department of Family Oral Health, Faculty of Dentistry, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300, Kuala Lumpur, Malaysia. Department of Restorative Dentistry, Faculty of Dentistry, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300, Kuala Lumpur, Malaysia. Department of Craniofacial Diagnostic and Biosciences, Faculty of Dentistry, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300, Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected], [email protected], [email protected], [email protected], [email protected] Dental pulp contains an extracellular matrix that provides the nutrients and nerve supply for dentin formation, and maintains healthy teeth. However, the pulp can be damaged by inflammation and infection caused by bacteria in caries formation. Tissue engineering (TE) is an alternative treatment to restore pulp tissue function. However, the study of using Wharton’s Jelly mesenchymal stem cell (WJMSC) that can provide the same functionality as the dentin-pulp complex is still limited. In this study, different concentrations of BMP-7 will be used on the Wharton’s Jelly mesenchymal stem cell (WJMSC), and cultured in decellularized human dental pulp (DHDP) to investigate the histology and viability of the regenerated pulp. To compare normal pulp (NP), decellularized pulp (DHDP), and WJMSC treated with BMP-7, cultured in DHDP (WJMSC + BMP-7 + DHDP) based on morphology and histological staining. After staining with hematoxylin-eosin (HE) and Masson’s trichrome, the tissues were observed under the inverted microscope. Cell proliferation of WJMSC + BMP-7 + DHDP were determined using Alamar blue to count the viable cells in the culture. Based on histological staining, there is no difference in the nuclei presence between WJMSC treated with 50ng/ml BMP-7 and 25ng/ml BMP-7. However, both treatments were better than those without treatment with BMP-7. WJMSC treated with 25ng/ml of BMP-7 showed higher cell viability compared to 50ng/ml of BMP7 and without BMP-7 treatment. This indicated that 25ng/ml of BMP-7 is an appropriate dose to enhance WJMSC proliferation in DHDP. Hence, the addition of BMP-7 can influence the performance of WJMSC and DHDP in revitalizing dental pulp for tissue engineering. Keywords: Bone morphogenetic protein 7, Wharton’s Jelly mesenchymal stem cells, Decellularized human dental pulp 71 1 2 3 1 1 2 3 3
Presenter’s ID: OD01 Category: Natural Product ANTIMICROBIAL ACTIVITY OF MALABARICONE C AGAINST ORAL BACTERIA Nur Fathihah binti Nordin , Syarifah Nur Syed Abdul Rahman , Wan Himratul Aznita Wan Harun , Nur'ain Salehen Department of Biomedical Science, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, Malaysia. Department of Oral and Craniofacial Sciences, Faculty of Dentistry, Universiti Malaya, Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected] The emergence of antibiotic resistance among oral bacteria emphasizes the importance of discovering new antimicrobial agents. This study aimed to investigate the antimicrobial activity of malabaricone C, from Myristica cinnamomea, against primary oral bacteria colonizers such as Streptococcus mitis and Streptococcus sanguinis, as well as a secondary colonizer, Streptococcus mutans, and mixed species. The antimicrobial and anti-biofilm potential of malabaricone C, was evaluated by preparing oral bacteria suspensions and subjected to disc diffusion, broth microdilution, crystal violet assay, MTT assay, Field Emission Scanning Electron Microscopy (FESEM) analysis and LIVE/DEAD BacLight Bacterial Viability Kits using fluorescence microscope. Malabaricone C showed significant inhibition zones against all species, with the highest inhibition observed against S. mitis (30.27 ± 1.97 mm), followed by S. sanguinis (12.71 ± 1.27 mm), mixed species (10.98 ± 0.85 mm) and S. mutans (10.56 ± 1.35 mm) in disc diffusion assay. Malabaricone C exhibited minimum inhibitory concentration (MIC) of 3.13 mg/ml against all species, and a minimum bactericidal concentration (MBC) of below 0.39 mg/ml for S. mitis and S. sanguinis, while 6.25 mg/ml for S. mutans and mixed species. The crystal violet and MTT assays revealed that malabaricone C caused a dose-dependent decrease in biofilm formation and cell viability respectively across all species. S. mitis exhibited the highest susceptibility to malabaricone C, while S. mutans showed the lowest susceptibility. Hence, it is shown that malabaricone C demonstrated potent antibacterial activity against oral bacteria, as evidence by significant inhibition zones, low MIC values, dose-dependent reduction in the crystal violet and MTT assays. These current findings highlight the potential of malabaricone C as a promising antimicrobial agent against oral pathogens. In addition, this study is still ongoing with current efforts focused on conducting FESEM analysis and live and dead staining to provide more comprehensive insight into the antimicrobial activity of malabaricone C. Further studies are also needed to explore its potential applications in oral health. Keywords: malabaricone C, antimicrobial activity, oral bacteria 72 1 2 1 2 2 1*
Presenter’s ID: OD02 Category: Natural Products THE ANTIMALARIAL ACTIVITY OF THE PEEL EXTRACT OF Garcinia mangostana AND THE COMPOUNDS, GONIOTHALAMIN FROM Goniothalamus tapis AND ACETOXYCHAVICOL ACETATE FROM Alpinia conchigera AGAINST THE MALARIA PARASITE, Plasmodium falciparum Nurul Amanina Binti Abdullah, Ahmad Firdaus Bin Ahmad Sukri School of Health Sciences, Health Campus, Universiti Sains Malaysia, Kelantan, Malaysia. *Corresponding author email address: [email protected], [email protected] Malaria is a severe and fatal disease caused by Plasmodium spp. and remains one of the leading global causes of morbidity and mortality. The emergence of drug-resistance P. falciparum in various countries has necessitated an effort to discover new antimalarial drugs targeting different pathways. The antimalarial activity of the peel extract of G. mangostana and the compound goniothalamin isolated from G. tapis have been reported in a few studies, offering insights into potential new antimalarial drug research. While acetoxychavicol acetate has been reported to possess various biological activities, its potential as an antimalarial agent has remained undiscovered. Therefore, this study aimed to fill this knowledge gap by investigating the antimalarial activity of acetoxychavicol acetate isolated from A. conchigera. The antimalarial potential of the peel extract, goniothalamin and acetoxychavicol acetate against the chloroquine-sensitive strain (3D7) of P. falciparum was assessed by using a malarial SYBR Green Ӏ fluorescence-based (MSF) assay. The peel extract and acetoxychavicol acetate exhibited promising antimalarial activity with an IC50 value of 12.49 and 10.08 μg/mL, respectively. Meanwhile, goniothalamin showed a moderate activity with an IC50 value of 17.98 μg/mL. A further investigation focused on the effect of acetoxychavicol acetate towards the pH of the mid trophozoite stage parasite’s digestive vacuole, employing a flow cytometry-based technique with fluorescein xx isothiocyanate-dextran (FITC-dextran) as a pH ratiometric probe. The results revealed a concentration-dependent increase in pH following acetoxychavicol acetate treatment, similar to the effect observed with the parasite treated with a standard proton pump inhibitor, concanamycin A. This suggests that acetoxychavicol acetate might have altered the digestive vacuole’s pH through the inhibition of V-type H+ -ATPase that regulates the acidification of the vacuole. Overall, this study provides crucial evidence of acetoxychavicol acetate capability as a promising antimalarial candidate. Keywords: Malaria, antimalarial drugs, acetoxychavicol acetate 73
Presenter’s ID: OD03 Category: Natural Products WOUND HEALING PROPERTY OF GREEN SYNTHESIZED ZINC OXIDE NANOPARTICLE, COPPER OXIDE NANOPARTICLE, AND ZINC/COPPER NANO COMPOSITE FROM Citrus sinensis PEEL EXTRACT ON HUMAN SKIN CELLS Jia Hong Chin, Sinouvassane Djearamane Department of Allied Health Sciences, Universiti Tunku Abdul Rahman, Kampar, Perak. *Corresponding author email address: [email protected]; [email protected] Wound management imposed a significant burden on global healthcare systems. Various established healing therapies come with drawbacks that hinder their efficiency in wound healing. Green synthesis of nanoparticles is gaining attention for their promising wound healing activities and lower toxicity than chemically synthesized nanoparticles. The objectives of this study are to synthesize zinc oxide nanoparticles (ZnO NP), copper oxide nanoparticles (CuO NP), and zinc/copper nanocomposites (ZnO/CuO NC) using Citrus sinensis peel and to characterize them, followed by accessing their wound healing property through wound scratch assay. UV-Visible spectroscopy (UV-Vis) analysis showed absorption peak of ZnO NP, CuO NP, and ZnO/CuO NC at 378 nm, 302 nm, and 376 nm, with energy bandgap at 3.73 eV, 2.71 eV, and 2.31 eV respectively. The ZnO NPs were spherical, while both CuO NP and ZnO/CuO NC exhibited irregular shapes. Additionally, ZnO NP exhibited a hexagonal wirzite crystalline structure, contrasting with the monoclinic crystalline shape of CuO NP, while ZnO/CuO NC displayed a combination of both structures. The crystalline size of ZnO NP, CuO NP, and ZnO/CuO NC was 20.27 nm, 12.40 nm, and 23.46 nm respectively. Fourier transform infrared spectroscopy (FT-IR) analysis revealed distinct peaks corresponding to Zn-O stretching in ZnO NPs, Cu-O stretching in CuO NP, and a combination of Zn-O and Cu-O stretching in ZnO/CuO NC. Wound scratch assay was performed using HaCat cells and the study results revealed the percentage of wound closure ranging from 37.26 to 56.46 % for CuO/ZnO NC, followed by 26.13 to 46.25 % for ZnO NPs, and 22.18 to 35.72 % for CuO NPs, at 24 h for the concentration ranging from 1 to 5 µg/mL respectively. However, all the tested concentration of NPs and NC showed complete (100%) wound healing at 48 h. Overall, ZnO/CuO NCs exhibited the greatest wound healing activity, followed by ZnO NP and CuO NPs. Keywords: Nanoparticles, Nanocomposite, Wound healing 74
Presenter’s ID: OD04 Category: Natural Products THE EFFECTS OF Plukenetia volubilis AGAINST MEMORY DEFICITS IN LIPOPOLYSACCHARIDE-INDUCED FEMALE RAT MODEL OF ALZHEIMER’S DISEASE Lee Choon Yung, Zuraidah binti Abdullah, Sabreena binti Safuan School of Health Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan, Malaysia. Alzheimer’s disease (AD) is an irreversible neurodegenerative disorder that causes decline in the six cognitive domains and affects the overall well-being of individuals afflicted with AD. Current medications for AD include lecanemab, donepezil, and memantine HCl. However, these drugs do not cure the disease and individuals may develop side effects such as headache and confusion with some developing heart failure and liver failure. This study investigates the effects of Plukenetia volubilis aqueous extract as an alternative treatment in treating AD in lipopolysaccharide (LPS) induced rat model. In silico analysis were performed to determine the viability of the phytochemicals to penetrate the blood-brain barrier (BBB) and exert effect on the CDK5/P25 complex associated with AD. Learning and memory were assessed using the Morris water maze (MWM) and novel object recognition (NORT). The blood pressure of the rats was assessed for blood pressurelowering effects. Toxicity studies were assessed by using haematoxylin and eosin (H&E) to study the effects of the aforementioned treatment on brain, liver, lung, and kidney. Results showed that LPS-induced rats treated with 500mg/kg P. volubilis aqueous extract group have the highest spatial learning in the trend of escape latency compared to other groups during the training phase in MWM, however, the test results showed statistically insignificant differences between groups. The LPS-induced rat treated with 500mg/kg P. volubilis aqueous extract group also shows lower mean blood pressure compared to normal group. Furthermore, H&E staining shows that the P. volubilis aqueous extract reduced the formation of neurofibrillary tangles (NFTs) to normal in LPS-induced brain, reduced inflammation in the lung proposing anti-inflammatory effects from the treatment whereas no toxicity effects were seen on liver and kidney. In conclusion, P. volubilis aqueous extract could exert improvement in learning and memory by reducing NFTs in AD brain, antiinflammatory effects, and blood pressure-lowering effects without causing toxicity effects in vital organs. Keywords: Alzheimer’s disease, Plukenetia volubilis, neurofibrillary tangles 75
Presenter’s ID: OD05 Category: Natural Product PRELIMINARY INVESTIGATION ON THE IMMUNOMODULATORY MECHANISM OF SANG HUANG (Phellinus linteus LiGNO-SH02) EXTRACT (xSH ) ON MACROPHAGES Phoebe Tee Yon Ern , Tan Chong Seng , Ng Szu Ting , Fung Shin Yee , Adeline Chia Yoke Yin School of Biosciences, Taylor’s University, No. 1 Jalan Taylor’s, 47500 Subang Jaya, Malaysia. LiGNO Biotech Sdn Bhd., No. 1, Jalan Perindustrian Balakong Jaya 2/2, 43300 Balakong Jaya, Malaysia. Department of Molecular Medicine, Universiti Malaya, 50603 Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected], [email protected] Extracts from the medicinal mushroom Phellinus linteus (PL) have been used in traditional medicine against various diseases and claimed to possess immunomodulatory properties, probably through their interaction with immune cells such as macrophages. Previous studies have reported varying results, with some suggesting its anti-inflammatory activities and others proposing its immune-boosting effects. Hence, the current study was carried out to further investigate the immunomodulatory properties of PL extract on macrophages. MTT assay was performed on RAW264.7 and phorbol-12-myristate 13- acetate (PMA)-differentiated THP-1 cells treated with 50 to 500 μg/ml of PL extract. PL extracts did not exhibit cytotoxic effects at concentrations of up to 300 μg/ml, thus, concentrations of 50, 150, and 300 μg/ml were chosen for further analysis. Macrophages were stimulated with lipopolysaccharide and treated with or without 50, 150, or 300 μg/ml of PL extract for 24 hours. Subsequent assays were performed to measure radical scavenging activities, nitric oxide (NO) production, reactive oxygen species (ROS) generation, and cytokine expression. The PL extract itself did not display DPPHscavenging activities, and showed no significant modulation of macrophage NO and ROS production. However, gene expression analysis revealed significant enhancement of inflammatory cytokines interleukin (IL)-1β, IL-6, and tumour necrosis factor (TNF)-α. Thus, despite the absence of radical-scavenging activities, the pro-inflammatory effect of the PL extract suggests its potential as an immune enhancer for use in conditions such as immuno-deficiency disorders and chemotherapy. Nevertheless, further studies should be performed to determine the underlying mechanisms behind the pro-inflammatory properties exhibited by PL. Keywords: Immunomodulatory, medicinal mushroom, Phellinus linteus 76 1 2 3 1 2 2 1* 3* TM
Presenter’s ID: OD06 Category: Natural Products BACTERICIDAL ACTION OF STINGLESS BEE HONEY AGAINST PATHOGENIC Escherichia coli Kang Chiao Sim Department of Allied Health Sciences, Universiti Tunku Abdul Rahman, Kampar, Malaysia. *Corresponding author email address: [email protected] Despite the antimicrobial activity of stingless bee honey has been well investigated, the mechanism underlying the bactericidal action against pathogenic bacteria is still understudied. Thus, the main objective of present study aimed to investigate the bactericidal action of stingless bee honey against Escherichia coli. The study started with the comparison of antimicrobial potency of Malaysian stingless bee honeys, the Geniotrigona thoracica and Heterotrigona itama honeys. Both agar well diffusion and broth dilution assays showed that G. thoracica honey exhibited superior inhibitory and bactericidal activities against both susceptible and multidrug-resistant strains of E. coli compared to H. itama honey, as evidenced by the formation of larger inhibition zones (7.0 – 12.0 mm), lower MIC and lower MBC values (6.25 – 12.5% w/v). Next, the bactericidal action of the G. thoracica honey on the morphological integrity of E. coli was investigated using Bradford assay, DiBAC4(3) and propidium iodide fluorescence staining methods and scanning electron microscopy (SEM). The outcomes showed that stingless bee honey exhibited stronger bactericidal activities than the membrane-targeting antibiotic polymyxin B, as reflected by the significantly higher protein leakage in a time-independent manner (p > 0.05). Higher fluorescence intensities were observed in stingless bee honey treated E. coli isolates, suggested the superior membrane depolarizing and permeabilizing capabilities of the honey. Remarkable membrane deformations and the appearance of elongated aseptate cells were also evidenced in SEM micrographs after E. coli was treated with the honey. The findings of this study suggested that the bactericidal effect of stingless bee honey was mainly attributed to its membrane alteration and even cell division inhibiting properties. Keywords: stingless bee honey, antibiotic resistant, bactericidal 77
Presenter’s ID: OD07 Category: Natural Products EVALUATION OF ANTIOXIDANT PROPERTIES AND CHEMICAL COMPOSITION OF SPIRULINA AND CHLORELLA EXTRACTS FOR FACE CREAM FORMULATION Dhanaluxmi Pillay Ramasawmy , Ming-Li Teoh , Sook Wah Chan , Nallammai Singaram School of Biosciences, Faculty of Health and Medical Sciences, Taylor’s University, Subang Jaya 47500, Malaysia. Clean Technology Impact Lab, Taylor’s University, Subang Jaya 47500, Malaysia. Food Security & Nutrition Impact Lab, Taylor’s University, Subang Jaya 47500, Malaysia. *Corresponding author email address: [email protected] Currently, with the rise of fame in the green market, the beauty and cosmetic industry aims to acquire sustainability while providing natural and non-toxic products to the public. Microalgae have garnered significant interest due to their sustainable status and the bioactive compounds they contain. These compounds show little to no risk of inducing skin allergenic effects and exhibit anti-inflammatory, anti-oxidative, anti-ageing, photoprotective and immunomodulatory properties. Therefore, this study aims to formulate face creams from Chlorella and Spirulina and to characterise their antioxidant and chemical properties. The bioactive compounds of each strain were extracted using three solvents, namely hexane, water, and ethanol. The antioxidant activity of the ethanolic and aqueous extracts was determined using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) tests while their phytochemical content was assessed using the total phenolic assay (TPC). The microalgae extracts were subjected to gas chromatography-mass spectrometry (GCMS) analysis. The face creams containing each strain’s ethanolic and aqueous extracts were formulated, and their properties were tested. Spirulina extract exhibited higher antioxidant activities (IC50 values of 0.70 ± 0.03 to 20.67 ± 1.21 mg/mL for DPPH and 6.22 ± 0.14 to 7.32 ± 0.07 mg/mL for ABTS) compared to Chlorella extract (IC50 values of 4.11 ± 0.26 to 37.07 ± 3.80 mg/mL for DPPH and 10.58 ± 0.22 to 47.07 ± 3.92 mg/mL for ABTS). For the TPC, at 6 mg/mL, the ethanol extracts of both Spirulina and Chlorella strains were the highest with reported values of 0.02 ± 0.00 mg GAE/g dry weight. GCMS results identified 39 compounds in the Spirulina extract compared to 44 in the Chlorella extract. The results indicated that the ethanol extract for both Chlorella and Spirulina strains possessed a significant quantity of bioactive compounds. The physical characteristics of the face creams formulated from these extracts were stable and optimal. Keywords: Chlorella, Spirulina, antioxidant properties, face cream 78 1 1,2* 1,2 1,3 1 2 3
Presenter’s ID: OD08 Category: Natural Products PHYTOCHEMICAL ANALYSIS AND ANTIBACTERIAL ACTIVITY OF Myristica fragrans FRUIT EXTRACTS AGAINST HUMAN PATHOGENIC BACTERIA Yong Darren Seow Tien , Nam Weng Sit Department of Allied Health Sciences, Faculty of Science, Universiti Tunku Abdul Rahman, Kampar, Perak, Malaysia. *Corresponding author email address: [email protected] The emergence of antibiotic resistant bacteria has become a clinical concern due to the ineffectiveness of conventional antibiotics in infection treatment. Myristica fragrans has been used in traditional medicine to treat infectious diseases. The objective of this study is to detect and quantify the phytoconstituents and to compare the antibacterial efficacy of different M. fragrans extracts against human pathogenic bacteria. Myristica fragrans extracts were obtained using sequential maceration technique. Nine phytochemicals were screened through a qualitative phytochemical test. The total phenolic content (TPC) and total flavonoid content (TFC) were determined using Folin-Ciocâlteu and aluminium chloride colourimetric assays, respectively. Colourimetric broth microdilution method was adopted to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of M. fragrans pericarp and seed extracts against nine bacterial strains. Phytochemicals such as alkaloids, carotenoids, flavonoids, glycosides, phenolics, quinones, saponins, terpenoids and tannins were detected in the extracts at variable degree. The ethyl acetate and ethanolic extracts of the seed had the highest TPC recorded at 46.719 ± 1.675 and 46.338 ± 3.388 μg GAE/mg extract, respectively. Whereas the seed ethanolic extract had the highest total flavonoid content recorded at 166.153 ± 22.799 μg QE/mg extract. The MIC of pericarp and seed extracts were recorded at 1.25 to 2.50 mg/mL and 0.16 to 2.50 mg/mL respectively, whereas the MBC of pericarp and seed extracts were recorded at 2.50 mg/mL and 1.25 to 2.50 mg/mL, respectively. The seed extracts exhibited stronger antibacterial activity against tested bacterial strains as compared to the pericarp extracts. Extracts which are semi-polar in nature, and with higher total phenolic and flavonoid contents exhibited greatest antibacterial activity. Gram-positive bacteria were more susceptible to the treatment of M. fragrans extract as compared to Gram-negative bacteria. The antibacterial efficacy of M. fragrans extracts did not differ significantly between the reference and clinical strain of Escherichia coli and Klebsiella pneumoniae. Keywords: Nutmeg, TPC and TFC, Antibiotic Resistant Bacteria 79 *
Presenter’s ID: OD09 Category: Natural Products THE EFFECTS OF Plukenetia volubilis ON DEHYDROEPIANDROSTERONEINDUCED POLYCYSTIC OVARY SYNDROME IN SPRAGUE DAWLEY RATS Muhammad Syarif Bin Zainal , Zuraidah Binti Abdullah, Sabreena Binti Safuan School of Health Sciences, Health Campus, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia. *Corresponding author email address: [email protected] Polycystic Ovary Syndrome (PCOS) is an endocrine disorder associated with various health issues including insulin resistance, obesity and an elevated risk of developing type 2 diabetes. This disorder is typically characterised by the presence of cysts in the ovaries, hyperandrogenism, infertility and menstrual cycle irregularities. The actual etiology of PCOS is still unclear, resulting in difficulty of planned treatment. Currently, no medication is available to specifically treat PCOS. Metformin is the go-to standard, but it only reduces PCOS symptoms without treating the underlying causes. Plus, side effects of metformin consumption include gastrointestinal symptoms such as nausea, vomiting and diarrhoea. The objective of this study is to investigate the effects of Plukenetia volubilis on PCOS-induced rats. 15 female Sprague Dawley Rats were divided into 5 groups of 3 animals each; normal, PCOS, 150 mg/kg metformin, 1000 mg/kg and 500 mg/kg P. volubilis. PCOS was chemically induced in rats using dehydroepiandrosterone (DHEA) intraperitoneally for 21 days. Treatments were given orally for 28 days after the induction period. At the end of treatment, serum hormonal profile (testosterone, estradiol, FSH and LH) was carried out. Ovary, liver and kidney were stained with haematoxylin and eosin, scored and analysed. PCOS induction was successful based on the significant testosterone level elevation (p=0.030) of PCOS group compared to normal. Upon treatment, elevation of corpus luteum number in 500 mg/kg (p=0.199) and 1000 mg/kg (p=0.071) P. volubilis groups could be observed compared to PCOS group. There were significant differences in estradiol level between PCOS group with normal (p=0.015) and 500 mg/kg P. volubilis group (p=0.001). Normal morphology of kidney and liver of P. volubilis groups suggested that P. volubilis was safe to be consumed. In conclusion, P. volubilis can reduce PCOS characteristics in rat models and further studies are needed to better understand P. volubilis’s mechanism in treating PCOS. Keywords: PCOS, P.volubilis, infertility 80 *
Presenter’s ID: OD10 Category: Natural Products BIOCHEMICAL COMPOSITION AND ANTIOXIDANT PROPERTIES OF CHLORELLA STRAINS FROM POLAR AND TROPICAL REGIONS Chloe Zi En Wong , Ming-Li Teoh , Sook-Wah Chan , Yin-Yin Ooi School of Biosciences, Faculty of Health and Medical Sciences, Taylor’s University, Subang Jaya 47500, Malaysia Clean Technology Impact Lab, Taylor’s University, Subang Jaya 47500, Malaysia Food Security & Nutrition Impact Lab, Taylor’s University, Subang Jaya 47500, Malaysia *Corresponding author email address: [email protected] With the rise of interest in acquiring sustainability in the pharmaceutical industry, there has been an emphasis on finding natural resources to replace the use of synthetic compounds used in products. Microalgae have garnered significant attention owing to their natural and sustainable capability to produce a diverse array of bioactive compounds. Therefore, this study aimed to screen for the possible biochemical composition and antioxidant properties of Chlorella strains from a tropical region (Chlorella UMACC 051 and Chlorella UMACC 038) and a polar region (Chlorella UMACC 250 and Chlorella UMACC 234). The microalgae cultures were grown for ten days. At the end of the experiment, the specific growth rate, chlorophyll-a content, carotenoid content, biomass, and biochemical composition such as carbohydrate, protein and lipid content were determined. In addition, the phytochemical properties were determined using a total phenolic assay, while the antioxidant activities were determined using 2,2-Diphenyl-1-picrylhydrazyl (DPPH). Of all Chlorella strains tested, the tropical Chlorella UMACC 051 showed the fastest growth rate and most biomass while the polar Chlorella UMACC 234 contained the highest pigment content and tropical Chlorella 038 had the highest total phenolic content. The biochemical composition analysis showed all strains to have a high lipid content ranging from 45.36 to 60.30 % Dry weight (DW) which does not correlate to the general expectation as microalgae is usually seen to have high protein contents, hence it is speculated that this is a form of adaptation or metabolic shift towards lipid production in these strains. All Chlorella strains exhibited a small amount of antioxidant activity (15.42 to 30.15 %) and total phenolic content ranging from 1.91 ± 0.04 to 4.43 ± 0.10 mg GAE/g DW. From the results, it can be concluded that polar Chlorella UMACC 234 has the most potential in containing significant amounts of bioactive compounds. Keywords: Chlorella, biochemical composition, antioxidant. 81 1 1,2* 1,3 1 1 2 3
Presenter’s ID: PA01 Category: Therapeutic and Diagnostic SCREENING FOR ANTIBIOFILM EFFECT OF PEPTIDE PAM-6 ON Pseudomonas aeruginosa Adeline Tiaw Yuen Shan, Yuen Hawk Leong Department of Allied Health Sciences, Universiti Tunku Abdul Rahman, Kampar, Perak *Corresponding author email address: [email protected], [email protected] Pseudomonas aeruginosa poses a significant health threat as it can cause acute infections through planktonic bacteria and transition into chronic infections by forming biofilms, which enhances resistance to conventional antibiotics. Recognizing the need for alternative antimicrobial strategies, recent research has introduced the potential of antibacterial peptides (ABPs). This study aimed to explore the efficacy of PAM-6 in inhibiting biofilm formation and eradicating established biofilms of Pseudomonas aeruginosa. Both biofilm inhibition and eradication assays were conducted. In the biofilm inhibition assay, microtiter plates were pre-coated with PAM-6 at concentrations ranging from 2 µg/mL to 256 µg/mL, followed by incubation with P. aeruginosa for 24 hours. Biofilm biomass was quantified using crystal violet staining, and inhibition percentages were calculated. For the biofilm eradication assay, the bacterium was allowed to form biofilms on the wells for 48 hours before treatment with PAM-6 at the same concentrations. Biofilm biomass was assessed using crystal violet staining, and the metabolic activity of bacterial cells was determined using the MTT assay. The results revealed that PAM-6 inhibited more than 50% of biofilm formation at a concentration of 8 µg/mL in the biofilm inhibition assay. In the biofilm eradication assay, concentrations of 8 µg/mL and 32 µg/mL resulted in the eradication of more than 50% of biofilm biomass, as determined by the crystal violet and MTT methods, respectively. Overall, PAM-6 demonstrated promising potential in reducing, inhibiting, and eradicating biofilmembedded bacteria of P. aeruginosa. The efficacy increased with higher concentrations of PAM-6, as evidenced by escalating biofilm inhibition and eradication percentages. Keywords: Antibacterial peptide (ABP), PAM-6, Biofilm 82
Presenter’s ID: PA02 Category: Therapeutic and Diagnostic EVALUATION OF CYTOTOXICITY OF CARBON MONOXIDE-RELEASING MOLECULE 2 (CORM-2) ON A549 LUNG EPITHELIAL CELLS Nafiessa Alya Mohd Najib , Ahmad Zaidi Ismail , Hanis Hazeera Harith Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. *Corresponding author email address: [email protected] Acute lung injury (ALI) is a respiratory disorder that is due to persistent lung inflammation. Lung epithelial cells contribute to ALI by releasing pro-inflammatory mediators upon exposure to inflammatory stimuli. A549 lung epithelial cell line has been primarily used to screen for agents with therapeutic potential against ALI. The antiinflammatory properties of carbon monoxide (CO) have led to the development of COreleasing molecules (CORMs). However, their therapeutic effect against ALI is yet to be fully elucidated. This study aims to determine the non-cytotoxic concentrations of CORM-2 and inactive CORM-2 (iCORM-2) in A549 cells for application in subsequent assays. CORM-2 solution was prepared by dissolving in DMSO, while iCORM-2 was prepared by leaving CORM-2 solution at room temperature for 24 hours to release CO and confirmed by FTIR spectroscopy. MTT cytotoxicity assay was carried out by treating cells with increasing concentrations of CORM-2 or iCORM-2 ranging between 1.56 to 200 µM and analyzed after 24 hours. At 200 µM, CORM-2 significantly decreased A549 cell viability by 38% compared to the untreated group, while concentrations at or below 100 µM maintained at least 80% viability. Similarly, iCORM-2 significantly reduced viability by 33% at 25 µM and above, with 12.5 µM or below showing minimal cytotoxicity. iCORM-2 showed heightened cytotoxicity at levels that are non-toxic for CORM-2 in A549 cells. This could be attributed to the modified chemical composition of the complex upon CO release, wherein DMSO substitutes CO forming Ru(Cl)2(DMSO)4, along with other complexes which may be toxic towards A549 cells. The concentrations considered non-cytotoxic for CORM-2 ranges from 1.56 to 100 µM, and from 1.56 to 12.5 µM for iCORM-2. Since iCORM-2 represents the control for CORM-2, concentrations between 1.56 and 12.5 µM will be applied to evaluate the antiinflammatory properties of CORM-2 in A549 cells. Keywords: CORMs, A549, acute lung injury 83 1 2 1 2 1*
Presenter’s ID: PA03 Category: Therapeutic and Diagnostic STUDY OF MECHANISMS OF ACTIONS OF ANTIBACTERIAL PEPTIDE PAM-6 ON Escherichia coli AND Pseudomonas aeruginosa Cheng Jia Ai, Yuen Hawk Leong Department of Allied Health Sciences, Universiti Tunku Abdul Rahman, Kampar, Perak. *Corresponding author email address: [email protected], [email protected] Antibiotic resistance is a global crisis that is mainly caused by the ineffectiveness of conventional antibiotics that possess only specific mechanisms, which lead to a higher possibility of mutational resistance to happen in the bacteria. One of the proposed solutions for this situation is that novel antibacterial agents that are effective and will not lead to resistance issue be developed. Antibacterial peptides (ABPs) are having the potential for becoming the novel antibacterial agents as they have multiple mechanisms and are unlikely to have the bacterial resistance be developed. A synthetic ABP known as PAM-6, was previously found to have bactericidal effect towards Escherichia coli and Pseudomonas aeruginosa. However, the actions of mechanisms of PAM-6 are yet to be elucidated. Hence, this study was conducted to screen for the membrane disruptive and DNA-binding effects of PAM-6 on E. coli and P. aeruginosa. For the determination of bacterial membrane disruptive effect of PAM-6, scanning electron microscopy (SEM) analysis was carried out to compare the surface morphology between the PAM-6-treated and untreated bacteria. Furthermore, electrophoretic mobility shift assay (EMSA) was also executed for the investigation of the DNA-binding effect of PAM-6 towards bacterial DNA. The bacterial DNAs were treated with different concentrations of peptide and incubated at room temperature for one hour before proceeding to gel electrophoresis. The experimental results shown that PAM-6 can cause bacterial outer membrane distortion in both bacteria as membranes blebbing were observed. Besides, PAM-6 also able to bind to the DNA of both bacteria as DNA retardations were observed in the EMSA at higher concentrations of PAM-6. These findings support the hypothesis that PAM-6 have multiple mechanisms of actions and may help to avoid the bacterial mutational resistance evolvement. Therefore, PAM-6 could be a potential novel antibacterial agent for the infections caused by E. coli and P. aeruginosa. Keywords: Antibacterial peptide, PAM-6, Mechanisms of actions 84
Presenter’s ID: PA04 Category: Therapeutic and Diagnostic INVESTIGATING THE ACTIVITY OF CRISPR/Cas9 RIBONUCLEOPROTEIN COMPLEX (RNP) TARGETING CD73 FOR CANCER IMMUNOTHERAPY Zi Jian Chong , Farid Nazer Faruqu Department of Biomedical Science, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, 50603, Malaysia. Department of Pharmacology, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, 50603, Malaysia. *Corresponding author email address: [email protected] CD73 is a GPI-anchored ecto-enzyme encoded by NT5E gene that mediates extracellular ATP metabolism to generate adenosine. CD73 is found to be overexpressed in various cancers, consequently increasing adenosine production, mainly driving an immunosuppressive tumour microenvironment thus promoting tumour survival. Therefore, CD73 has emerged as a novel immune checkpoint target with the potential to create a more immunostimulatory tumour microenvironment upon its inhibition. Currently, drugs developed against CD73 under clinical trials consist of small molecule inhibitors and monoclonal antibodies which only offer transient inhibition. The CRISPR/Cas9 system however can provide a permanent and precise gene editing. Thus, this study is aimed to synthesize a functional CRISPR/Cas9 RNP complex targeting CD73 for cancer immunotherapy. Firstly, 3 sgRNAs targeting CD73 (sgCD73_1, _2 & _3) are designed from the N-terminus, central and C-terminus of the NT5E gene, respectively. Subsequently, the sgCD73 is cloned into a Cas9-expressing plasmid (pX330). Next, the sgCD73-pX330 plasmid is transfected into HEK293T cells to determine its gene editing activity. Based on the in-silico gene editing efficiency prediction, it shows that sgCD73_1 which is designed near the N-terminus of NT5E gene would give the best CD73 knockout efficiency. However, the gene knockout analysis performed on genomic DNA extracted from HEK293T cells following transfection with sgCD73-px330 plasmid using ICE analysis tool showed that only sgCD73_2 is the only sgRNA effective to knockout CD73. To conclude, a CRISPR/Cas9 targeting CD73 has been successfully designed and validated. Given that CRISPR/Cas9 is a form of biologic and therefore suffers from delivery limitations in vivo, we will next clone work on having this CD73-targeting CRISPR/Cas9 system loaded into and delivered via extracellular vesicles (EVs) as a more efficient means to deliver CRISPR/Cas9 in vitro and in vivo for cancer immunotherapy. Keywords: CD73, Cancer, CRISPR/Cas9 85 1 2 1 2*
86 Presenter’s ID: PA05 Category: Therapeutic and Diagnostic MOLECULAR DETECTION OF MICROSPORIDIA SPP. AMONG ROHINGYA SCHOOL CHILDREN IN SELANGOR Muhammad Fitri Hakimi Bin Zulkifli , Nur Raihana Ithnin , Nurshahira Sulaiman Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia. Department of Medical Microbiology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia. *Corresponding author email address: [email protected], [email protected], [email protected] Microsporidia are obligate intracellular spore-forming protozoan parasites, which over 17 species have been discovered to infect humans. Two species that are commonly reported as the etiological agents in humans are Enterocytozoon bieneusi (E. bieneusi) and Encephalitozoon intestinalis (E.intestinalis). In Malaysia, infection cases of microsporidiosis have been reported among the high-risk groups of hospitalised patients and the Orang Asli community in rural areas. However, recent concern has emerged for the Rohingya community in Malaysia for their susceptibility to contract with emerging disease. As a marginalised community in Malaysia, there is less concern regarding their health status which resulted in the neglect of health surveillance in the Rohingya community especially the children who are prone to contract infectious diseases as well as parasitic infection. In order to diagnose microsporidiosis in Rohingya school children, a molecular approach has been used to facilitate the detection of microsporidia up to the species level. This study aims to determine the molecular epidemiological characteristics of microsporidia among Rohingya school children in Selangor. A total of 91 stool samples have been collected from Rohingya school children in Selangor and undergo DNA extraction followed by optimization of the PCR amplification. The multiplex Polymerase Chain Reaction (PCR) technique was performed to detect E. bieneusi and E.intestinalis in Rohingya children stool samples. Later, the amplified DNA was visualised onto gel electrophoresis for detection of E. bieneusi and E.intestinalis at 607 bp and 570 bp respectively. Preliminary results indicated positive presence of E. bieneusi and E.intestinalis among the samples with higher infection of E.intestinalis compared to E. bieneusi. Molecular approaches in detection of microsporidiosis prove to be effective in this study and the presence of E. bieneusi and E.intestinalis in this target population can be related to their hygienic lifestyles and exposure to their environment condition. There are still challenges to diagnose for microsporidiosis using molecular technique. Therefore, in order to achieve comprehensive understanding in detection of microsporidiosis, a thorough molecular studies of microsporidia species is required to improve the accuracy and sensitivity of this diagnosis. Keywords: Microsporidia, Polymerase Chain Reaction (PCR), Molecular Detection 1 2 1 2 1
Presenter’s ID: PA06 Category: Therapeutic and Diagnostic SUITABILITY OF HIGH-RESOLUTION MELT (HRM) ANALYSIS FOR SINGLE NUCLEOTIDE POLYMORPHISM DETECTION IN THE PATTANI MALAY POPULATION Zuliana Anisah Zulkifli , Mat Ghani Siti Nor Assyuhada , Nuur Athirah Mohd Daud , Noorzalifah Mazuki , Nurul Atika Musa , Sharifah Nany Rahayu Karmilla Syed Hassan , Ayunni Zainul Bahri , Naam Bahjat Ahmed Adeeb , Nik Norliza Nik Hassan, Nazihah Mohd Yunus , Mohamad Ros Sidek , Mohamad Zain Musa , Muhammadfahmee Talek , Farid Mat Zain , Numan Hayimasae , Md Salleh Yaapar , Bin Alwi Zilfalil Human Genome Centre, School of Medical Sciences, Health Campus, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan. School of Health Sciences, Health Campus, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan. Malaysian Node of the Human Variome Project, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan. Institute of the Malay World and Civilization, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor. Faculty of Nursing, Prince Songkla University, Pattani Campus, 94000 Pattani, Thailand. Center for Arabic Language and Islamic Civilization, Faculty of Islamic Studies, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor. Malay and Malay Studies Program, Faculty of Humanities and Social Sciences, Prince of Songkla University, 181, M.6, Charoen Pradit Rd., T.Rusamilae, A.Muang, Pattani, 94000 Thailand. School of Humanities, Universiti Sains Malaysia, 11800 Pulau Pinang. Faculty of Medicine, Universiti Sultan Zainal Abidin, Medical Campus, Jalan Sultan Mahmud, 20400, Kuala Terengganu, Terengganu. * Corresponding author email address: [email protected] This study investigates the genetic polymorphisms of the Pattani Malay population, emphasizing the distribution of genetic variations and their potential implications for their health and susceptibility to region-specific diseases. The study utilizes T-ARMS PCR, High-Resolution Melt (HRM) analysis, and Sanger sequencing to explore genetic traits and variations within the population. The HRM method effectively genotyped samples for the SNP rs961378, revealing three distinct variants, predominantly exhibiting a heterozygote genotype with genotype frequency of 0.60. Sanger sequencing verified the genetic traits of variants classification from HRM analysis, which include heterozygous, homozygous T and homozygous C. The study adhered to Hardy Weinberg Equilibrium with chi-squared value 0.40, providing insights into the distribution of genotypes. However, the limitation of a small sample size with 10 samples requires caution in interpretation, as it may lead to false positives and lack of representativeness. Validation in a larger sample size is recommended to ensure the reliability and generalizability of the findings. The study identifies challenges associated with the T-ARMS PCR method, emphasizing the need for improvements in primer design and contamination prevention. Despite offering valuable insights, the study highlights the importance of cautious interpretation due to the limitations. It suggests further research involving a larger, more diverse sample size and methodological enhancements to establish a better understanding of genetic associations in the Pattani Malay population, contributing to broader knowledge in the field. Keywords: T-ARMS PCR, HRM analysis, Sanger sequencing, Genotype frequency 87 1 2 3 4 5 6 7 8 9 1* 1,2 8 1 3 3 3 5 6 7 1 4 1 3 3,9 2 1
Presenter’s ID: PA07 Category: Therapeutic and Diagnostic THE STUDY OF NUDT15 GENOTYPES IN MALAYSIAN PATIENTS WITH CROHN’S DISEASE Huei Yiun Lee, Sheh Wen Kuan, Suat Moi Puah, Kek Heng Chua, Boon Pin Kee Department of Biomedical Science, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected] Crohn’s disease (CD) is a type of inflammatory bowel disease that leads to chronic inflammation of the gastrointestinal tract. Its prevalence has increased rapidly among the Asian populations recently, including Malaysia. Azathioprine, an immunosuppressive thiopurines drug, is commonly used in CD treatment. It suppresses the immune response by incorporating active metabolites into lymphocytes DNA to induce apoptosis. Genetic polymorphisms in Nudix hydrolase 15 (NUDT15) gene were recently discovered and consistently reported among Asians to be associated with impaired thiopurine metabolism, increasing the risk of adverse effects during thiopurine treatment. The study of NUDT15 variations in Malaysian population is lacking. Therefore, this study aims to determine the frequencies of NUDT15 genotypes in the Malaysian CD (N = 138) and control (N = 276) cohorts. The genotyping of four main polymorphisms in NUDT15 gene (rs746071566, rs186364861, rs116855232 and rs147390019) that contribute to reduced metabolising activities of NUDT15 was carried out using conventional PCR and tetra-primer amplification refractory mutation system (ARMS)-PCR approaches. In this study, all four polymorphisms were found among the Malaysian population (MAF: 0.0012 - 0.0978). A total of 9.78% of the patients and 11.41% of the control cohort carried the metabolismreducing NUDT15 alleles (NUDT15*2-NUDT15*6). Amongst, majority of them carried NUDT15*3 (7.49%) and NUDT15*2 (2.29%) alleles. Notably, NUDT15*4-*6 alleles are rare among Malaysian population (<1%). Based on their diplotypes, 19.32% is expected to exhibit intermediate activity (*1/*2, *1/*3, *1/*4 and *1/*6), whereas 1.21% is expected to exhibit low activity (*2/*3, *2/*5, *2/*6 and *3/*3). In conclusion, the NUDT15 genetic variations present at a substantial level (~10%) among the Malaysian population and it is expected that 20% of Malaysian population are at risk of developing thiopurine-related adverse effects. This warrants the need for thiopurine-related toxicity prediction screening prior to thiopurine treatment to reduce the risk of adverse effects among the Malaysian population. Keywords: NUDT15, Azathioprine, Crohn’s disease 88 *
Presenter’s ID: PA08 Category: Therapeutic and Diagnostic CHARACTERIZATION OF HUMAN MICROGLIAL MODEL INVOLVED IN METHAMPHETAMINE-INDUCED NEUROINFLAMMATION Tham Xuan Yao , Yang Kun , Lam Kok Wai , Tham Chau Ling Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia. Faculty of Pharmacy, Universiti Kebangsaan Malaysia. The abuse of a highly addictive drug, methamphetamine (Meth) due to the rewarding effects of elevated dopamine in the brain has caused serious neurological disorders. The production of proinflammatory cytokines acts as a hallmark of microglial activation. BV-2 cells, a murine microglial cell line, have been used as the main model to study Methinduced neuroinflammation. Human microglial clone 3 (HMC3) cells have been reported as a good model by exhibiting original antigenic properties and microglial markers, however, have not been employed to study Meth-induced neuroinflammation. This study aims to establish a Meth-induced neuroinflammation model using HMC3 cells by identifying the non-cytotoxic concentration of Meth that will induce neuroinflammation. HMC3 cells were treated with Meth concentrations (0-1000 µM) along with lipopolysaccharide (LPS) to induce neuroinflammation. MTT (3-(4,5-Dimethylthiazol-2- yl)-2,5-diphenyltetrazolium bromide) assay was performed to identify the non-cytotoxic concentrations of Meth. The degree of neuroinflammation induced by Meth and LPS based on nitrite production and level of interleukin (IL)-6 was determined using Griess assay and enzyme-linked immunosorbent assay (ELISA) respectively. All Meth concentrations were not significantly toxic to HMC3 cells. Both Meth/LPS-induced HMC3 cells failed to increase nitrite production, indicating that nitric oxide (NO) was not inducible by Meth and LPS. However, IL-6 level was significantly increased in Meth/LPSinduced HMC3 cells at all tested concentrations. Previous studies have proven that LPSinduced BV-2 cells significantly produced nitrite and IL-6, in contrast with HMC3 cells that only produced IL-6. Similar to HMC3 cells, Meth-induced BV-2 cells only showed an increase in IL-6 level but not nitrite. These findings have partially established that HMC3 cells may be a suitable human microglial model to study Meth-induced neuroinflammation, however, its gene and cytokine profiles should be further examined to fully establish the model. Keywords: methamphetamine (Meth), neuroinflammation, human microglial clone 3 (HMC3) cells 89 1 2 1 1 2 1
Presenter’s ID: PA09 Category: Therapeutic and Diagnostic IMMUNOHISTOCHEMISTRY (IHC) EXPRESSION OF ENDOTHELIAL CELL SPECIFIC MOLECULE-1 (ESM-1) AS PREDICTIVE BIOMARKERS IN TRIPLE NEGATIVE BREAST CANCER (TNBC) PATIENTS Amar Harris Arifin , Siti Norasikin Mohd Nafi Biomedicine Programme, School of Health Sciences, Universiti Sains Malaysia, Health Campus. 16150 Kubang Kerian, Kelantan. Histopathology & Cytology Laboratory, School of Medical Sciences, Universiti Sains Malaysia, Health Campus. 16150 Kubang Kerian, Kelantan. *Corresponding author email address: [email protected] Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer characterized by the absence of both estrogen (ER) and progesterone (PR) receptors, as well as the lack of human epidermal growth factor receptor 2 (HER2) expression. Despite its clinical relevance, TNBC lacks targeted therapeutics necessitating a better understanding of its molecular features. Endothelial Cell- Specific Molecule-1 (ESM-1) is a proteoglycan largely produced by endothelial cells participating in angiogenesis, tumor growth and metastasis by providing essential oxygen and nutrients essential for tumor cell growth. Due to its role in tumor growth, there is considerable interest in exploring endothelial cell-specific molecule-1 (ESM-1) as a potential predictive biomarker for TNBC aggresiveness. Therefore, the study aims to examine the expression of endocan of ESM-1 in TNBC utilizing immunohistochemistry (IHC). The study utilizes a cohort of 36 TNBC patient samples, analyzing ESM-1 expression patterns in tumor tissues. Immunohistochemical staining assesses the presence and localization of ESM-1 within the tumor microenvironment. Additionally, correlations between ESM-1 expression and histological features, including patient outcomes and disease progression explored. Preliminary findings indicate a distinctive pattern of ESM-1 expression in TNBC, with 52.8% of TNBC showing high level of IHC ESM-1 expression, suggesting its potential involvement in the pathogenesis of this aggressive breast cancer subtype. Furthermore, correlations between ESM-1 expression and clinical parameters, specifically tumor sizes, grades, stages and lymphovascular invasion shed light on its predictive value. These findings contribute to the growing understanding of TNBC biology and may inform the development of targeted treatments or prognostic markers. Moreover, this study enhances our understanding of the molecular landscape of TNBC, potentially paving the way for novel therapeutic strategies to improve patient outcomes in this challenging breast cancer subtype. Therefore, ESM-1 holds promise as the potential predictive biomarker for TNBC. Keywords: Triple-negative breast cancer (TNBC), Endothelial Cell-Specific Molecule-1 (ESM-1), Immunohistochemistry (IHC) 90 1 2 1* 2
Presenter’s ID: PB01 Category: Therapeutic and Diagnostic EXPLORING THE EFFECTS OF ZERUMBONE ON THE EXPRESSION OF INVADOPODIA-RELATED PROTEINS IN COLON CANCER CELLS: A MOLECULAR INVESTIGATION Nur Izzah Muhammad Kamil, Nur Fariesha Md Hashim, Nurul Akmaryanti Abdullah, Noraina Muhammad Zakuan* Physiology Laboratory, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia. *Corresponding author email address: [email protected] Colorectal cancer (CRC) ranks as the third most prevalent cancer and the second leading cause of cancer deaths due to its metastatic potential. Treatments like radiotherapy and chemotherapy are compromised due to tumor-induced hypoxic microenvironments, fostering cancer cell invasiveness. Matrix Metalloproteinase 2 (MMP-2), cortactin, and Flightless I (FliI) are pivotal in the invadopodia formation which is crucial for cancer cell metastasis. Zerumbone (ZER) exhibits anticancer properties by reducing migration and invadopodia formation in HCT116 cells under normoxia and hypoxia conditions. However, studies on the effects of ZER on invadopodia-related protein in hypoxia-induced colon cancer cells are limited. The study aims to explore the effects of ZER on MMP-2, cortactin, and FliI expressions in hypoxia-induced HCT116 colon cancer cells. HCT116 cells were treated with ZER at IC20 (16 µM) and IC50 (35 µM) under normoxia, and at IC20 (20 µM) and IC50 (44 µM) under hypoxia for 24 hours. Western Blot Analysis was performed, followed by two-way ANOVA and Tukey post hoc test. HIF-1α is detected in the control group, IC20, and IC50 of ZER under hypoxia. MMP-2 expressions decreased in IC20 and IC50 of ZER under hypoxia compared to normoxia and the control groups. At IC20 of ZER, cortactin and FliI levels are similar between hypoxia and normoxia groups. However, at IC50 of ZER, there is a slight increase in cortactin and FliI levels in the hypoxia compared to the normoxia groups. HIF-1α is a key protein involved in cellular adaptation to hypoxia conditions. HIF-1α detection confirms effective induction by DMOG, suggesting a hypoxia model was established. No significant difference in MMP-2, cortactin, and FliI expressions was observed in cells treated with different concentrations of ZER. However, a trend of reduced MMP-2 expressions suggests ZER’s potential in inhibiting invadopodia formation, consistent with previous study. ZER demonstrates no significant effects on MMP-2, cortactin, and FliI expression levels. Keywords: Zerumbone, invadopodia-related proteins, hypoxia 91
Presenter’s ID: PB02 Category: Non-Communicable Disease MELATONIN REGULATES PYROPTOSIS BY TARGETING THE CASPASE1/CASPASE-4/CASPASE-5/GSDMD/HMGB1 PATHWAY IN HEP G2 CELL Yong Qiao Ru School of Health Sciences, International Medical University, Kuala Lumpur, Malaysia. *Corresponding author email address: [email protected] Hepatocellular carcinoma is the sixth most diagnosed cancer, and the fourth leading cause of cancer-related death in the world. Melatonin has beneficial properties such as chemopreventive, oncostatic and tumor inhibitory action. Pyroptosis is characterized by the activation of caspases and the formation of gasdermin channel. This study aims to investigate the melatonin-induced pyroptotic cell death in HepG2 cells by targeting caspase-1/caspase-4/caspase-5/GSDMD/HMGB1 pathway. HepG2 cells were cultured with DMEM medium with 10% FBS under 5% CO2 and treated with melatonin in different concentrations. Cell viability was measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5- Diphenyltetrazolium Bromide (MTT) assay. Cellular structures were stained by using Hoechst 33342 and propidium iodide (PI) dyes. Protein expressions were measured by Western blot analysis. Melatonin significantly (p<0.05) decreased the cell viability in a dose-dependent manner. The treated cells had poor morphology with shrunken size and small nuclei. No nuclear fragmentation was observed. Caspase 1, caspase 4, caspase 5, HMGB1 and GSDMD protein expressions decrease in a dose-dependent manner if compare with control. Melatonin has reduced the cell viability of Hep G2 cells due to its ability to induce cytotoxicity. Some cells swelled, indicating that the cells are undergoing pyroptosis. The production of caspases and GSDMD decrease in the dose-dependent manner, showing that melatonin inhibits pore formation and pyroptosis in HepG2 cell lines. The concentration of HMGB1 decreases in a dose dependent manner due to the anti-inflammatory action of melatonin. Melatonin induces cytotoxicity in HepG2 cells via regulating pyroptosis in a dose-dependent manner by targeting caspase 1/caspase 4/caspase 5/GSDMD/HMGB1 pathway. Keywords: Liver cancer, melatonin, pyroptosis 92
Presenter’s ID: PB03 Category: Non-Communicable Disease ANTICANCER PROPERTIES OF Limosilactobacillus reuteri 29B STRAIN IN HELA CELL LINE Leong Khei Min , Leslie Than Thian Lung , Elysha Nur Ismail Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor 43400 UPM, Malaysia. Department of Medical Microbiology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor 43400 UPM, Malaysia. *Corresponding author email address: [email protected] Cervical cancer has been ranked as one of the most common cancers worldwide with high incidence and mortality rate (54.56% as of 2018). Several lactic acid bacteria had been proven to exert anticancer effects on different types of cancer. Limosilactobacillus reuteri, a potential anticancer source, has been proven to stimulate antiproliferation and apoptotic cytokines such as TNF- and IFN- in cancerous cells. In recent research, newly isolated Limosilactobacillus reuteri 29B (L29B) from anogenital region has not been studied for its anticancer properties. Previously, it has been documented for its antimicrobial activity in in vitro and in vivo models. This study aims to assess the anticancer properties of L29B using HeLa cancer cells. Anticancer properties of L29B which were isolated from healthy Malaysian women vagina were studied using L29B cellfree supernatant (CFS) on the HeLa cell line. Cell-free supernatant of L29B was prepared by centrifugation of bacterial culture concentration OD600=0.1. The results were evaluated via proliferative, apoptotic and reactive oxygen species (ROS) assay. The CFS exhibited inhibitory effects of cancer cell growth via MTT assay after 24 hours treatment (p<0.05). Induction of apoptosis was caused by an increment in nitric oxide (NO). Higher levels of NO were measured in CFS treated cells. L29B CFS was able to prevent proliferation of HeLa cells by inducing oxidative stress on the cells. Proposed mechanism of action by researchers has included secretion of metabolites such as short-chain fatty acid, butyrate, bacteriocin and triggering of pro-apoptotic pathways of cancerous cells. Further studies needed to be conducted to assess the specific pathway or molecules of L29B postbiotic. Overall, CFS of L29B has been tested and proven as one of the potential supplements to prevent cervical cancer by the efficacy shown on human cervical cancer cell lines. Keywords: anticancer, postbiotics, HeLa cells 93 1 2 1* 2 1
Presenter’s ID: PB04 Category: Non-Communicable Disease UNLOCKING THERAPEUTIC INSIGHTS: CYTOTOXIC EVALUATION OF MALAYSIAN AGARWOOD LEAF EXTRACTS ON MELANOMA CELLS Ashlyn Yau Wen Ning , Looi Chung Yeng School of Biosciences, Taylor’s University, 47500 Subang Jaya, Selangor Darul Ehsan, Malaysia. Digital Health and Medical Advancement Impact lab, Taylor’s University, 47500 Subang Jaya, Selangor Darul Ehsan, Malaysia. *Corresponding author email address: [email protected] Melanoma is a highly lethal disease with increasing incidence rates. Natural anticancer agents are of great research interest to circumvent issues from current treatment strategies, such as adverse effects, chemoresistance and unaffordability. Aquilaria spp., commonly known as agarwood trees, yield natural compounds steeped in traditional and cultural practices, notably their leaves’ usage in health teas. This study aimed to evaluate the cytotoxicity of Aquilaria spp. methanolic leaf extracts on murine melanoma cells (B16-F10) and human keratinocytes (HaCaT). The leaves from two of the most common Aquilaria trees, Aquilaria malaccensis and Aquilaria sinensis, were extracted using maceration in methanol, then dried through a rotary evaporator, yielding dried crude methanolic extracts. MTT assay and real-time cell analysis (RTCA) were conducted to evaluate the cytotoxicity of the crude methanolic extracts through determination of the IC50 values after treatment for 24, 48 and 72 hours. The MTT assay indicated cytotoxicity of the extracts towards B16-F10 cells (IC50 ranging from 45.920 to 110.425 µg/mL). The extracts were however cytotoxic towards HaCaT cells at all time points. RTCA provided real-time data, showing immediate effect towards B16-F10 cells. Microscopic analysis of the cells after treatment indicated cell death. The results showed A. malaccensis to be slightly more cytotoxic than A. sinensis against B16-F10 cells, with the optimal candidate for treatment identified to be A. malaccensis at the 24-hour time point due to less pronounced toxicity against HaCaT cells, and the IC50 values towards B16-F10 cells being 56.028 µg/mL and 63.536 µg/mL respectively from the MTT assay and RTCA. The cytotoxicity of Aquilaria spp. leaf extracts on melanoma cells and keratinocytes was indicated in the study, emphasizing the importance of further studies into potential applications in cancer therapeutics, potentially providing therapeutic insights to mitigate issues with current treatment strategies. Keywords: Aquilaria malaccensis, Aquilaria sinensis, Melanoma 94 1 2 1 1,2*
Presenter’s ID: PB05 Category: Non-Communicable Disease THE CYTOTOXIC EFFECTS OF MALAYSIAN TUALANG ROCK HONEY AND TRIGONA HONEY ON MDA-MB-231 BREAST CANCER CELL LINE Kelly Chin Yu Han , Siti Norasikin Mohd Nafi , Nor Hayati Othman , Nik Norliza Nik Hassan Department of Biomedical Sciences, School of Health Sciences, Universiti Sains Malaysia, Health Campus, 16150 Kubang Kerian, Kelantan, Malaysia. Department of Pathology, School of Medical Sciences, Universiti Sains Malaysia, Health Campus, 16150 Kubang Kerian, Kelantan, Malaysia. *Corresponding author email address: [email protected] Tualang Rock honey and Trigona honey are two types of honey often found in Malaysia. Tualang Rock honey is obtained from Apis dorsata bees, while Trigona honey is produced from stingless Trigona bees. These honey subtypes include bioactive compounds with reported cytotoxic effects against various cancer cell lines, including breast cancer. The objective of this study was to assess the cytotoxic effects of Tualang Bitter Rock (TBR) honey, Tualang Sweet Rock (TSR) honey, and Trigona honey on the triple-negative breast cancer (TNBC) cell lines MDA-MB-231, using the MTT test. The MDA-MB-231 cell lines were cultured in complete Dulbecco's Modified Eagle Medium (DMEM) containing 100 units/mL of penicillin and 100 mg/mL of streptomycin, supplemented with 10% Fetal Bovine Serum (FBS), and maintained in humidified 37 °C incubator with 5% CO2. Cells were seeded at different densities and incubated for 24 hours to reach 70–80% confluence for all experiments. Then, cells were incubated at varying w/v concentrations of 0.01, 0.05, 0.1, and 0.5% w/v for 24 and 72 hours. Cell viability after incubation with the honey was determined by MTT assay. Trigona honey, TBR honey, and TSR honey showed no significant cytotoxic effects on MDA-MB-231 cells at various concentrations following a 24-hour treatment. Incubation to Trigona honey for 72 hours resulted in a significant reduction in cell viability, starting from lower to higher concentrations. After 72 hours of incubation, TSR honey exhibited a significant cytotoxic effect at a lower concentration of 0.10% w/v, but TBR honey demonstrated a significant cytotoxic effect at a higher concentration of 0.50% w/v. Our tested honey was shown to effectively inhibit the progression of TNBC cell lines when used at greater doses over a longer period of time. This study emphasizes the potential significance of honey in preventing the advancement of TNBC. Keywords: Tualang Rock honey and Trigona honey, breast cancer, cytotoxicity 95 1 2 1 2* 1 2
Presenter’s ID: PB06 Category: Non-Communicable Disease ANTI-OBESITY EFFECT OF DAIDZEIN ON METABOLIC-SYNDROME SPRAGUE DAWLEY RATS ACCOMPANIED WITH BENIGN PROSTATIC HYPERPLASIA Maizatul Harnani Mazlan , Siti Saleha Masrudin , Hasni Idayu Saidi , Nur Izzah Athirah Razwan Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. Department of Human Anatomy, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. *Corresponding author email address: [email protected]; [email protected] Scientific evidence has indicated that obesity in males contributes to the development of benign prostatic hyperplasia (BPH). It is due to high oestrogen levels in obese individuals, causing overstimulation activity of oestrogen receptor alpha. Since obesity and BPH involve oestrogen activity, research has been done on using daidzein as a treatment to treat both conditions individually, but not when they happen simultaneously. Therefore, this study is done to determine the anti-obesity effect of daidzein on metabolic syndrome rats accompanied with BPH. This is a 12-week study where 42 male Sprague Dawley rats are randomly assigned into 7 groups (n=6): normal control group, BPH control group, high-fat diet (HFD) control group, HFD-BPH control group, HFD-BPH-finasteride group (1 mg/kg), HFD-BPH-daidzein group (10 mg/kg), HFD-BPH-daidzein group (100 mg/kg). From week 9 until week 12, rats are injected subcutaneously with testosterone propionate to induce BPH, while the assigned treatments are given by oral gavage. At the end, all rats are euthanized, and targeted samples are collected. There are no significant differences in body weight and prostate index, but adipose tissue weights of the daidzeintreated rats showed significant differences compared to the HFD-BPH control group. Besides, daidzein-treated rats show a reduction in adipocyte size and prostate tissues when compared to the HFD-BPH control group. Although the food consumption and body weights of normal-fed rats and HFD-fed rats look similar, more adipose tissues are deposited in HFD-fed rats. This is due to different fat contents in the food. The targeted effects of daidzein are seen more clearly in the histopathology of adipocytes and prostates. Hence, relying solely on the body weight and prostate weight may lead to an unreliable finding of treatment. Daidzein does have potential for exhibiting anti-obesity and antiproliferative prostate effects on obese-BPH rats. Keywords: benign prostatic hyperplasia (BPH), high-fat diet (HFD), daidzein 96 2 1 1 2 1 2
Presenter’s ID: PB07 Category: Non-Communicable Disease THE THERAPEUTIC POTENTIAL OF Inonotus obliquus, Antrodia camphorata, AND Phellinus linteus MUSHROOM EXTRACTS ON BREAST CANCER CELLS Thiiben A/L Krishnan Sami , Looi Chung Yeng School of Biosciences, Taylor’s University, 47500 Subang Jaya, Selangor Darul Ehsan, Malaysia. Digital Health and Medical Advancement Impact lab, Taylor’s University, 47500 Subang Jaya, Selangor Darul Ehsan, Malaysia. *Corresponding author email address: [email protected] Breast cancer is known to be the most common cancer globally, with over 2 million cases and 670,000 deaths in the year 2022. Despite advances in breast cancer therapeutics, it still remains a major global concern. The current anticancer chemotherapeutic agents are associated with systematic toxicities, posing several side effects and clinical management complications. Cancer therapeutics based on natural substances have recently gained traction due to their efficacy and low toxicity. The present work focused on studying the therapeutic effects of Inonotus obliquus, Antrodia camphorata and Phellinus linteus (Sang Hwang) mushroom extracts on MCF-7 breast cancer cells and their respective toxicities on HaCaT epidermal keratinocytes by determining the half maximal inhibitory concentration (IC50) values. The cell viabilities were measured at 24, 48 and 72-hour time points using 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Further analysis was carried out for MCF-7 cells using Real-time Cell Analysis (RTCA) to verify the IC50 values obtained from the MTT assay and study the growth patterns of the cells exposed to the treatments. The results obtained from both assays revealed that all 3 mushroom extracts exerted cytotoxic effects on MCF-7 cells in a dose-dependent manner with Inonotus obliquus being the most potent, followed by Antrodia camphorata and Phellinus linteus, with distinct growth patterns obtained from RTCA. However, toxicity was observed at all the time points for HaCaT cells in all the treatments. In this study, A. camphorata appeared as the optimal candidate at the 24-hr time point with IC50 values of 325.00 and 225.52 ug/mL in MTT assay and RTCA respectively, where it recorded significant cytotoxicity on MCF-7 cells with minimal toxicity on HaCaT cells. This study yields preliminary data to demonstrate the promising therapeutic potential of medicinal mushrooms to possibly revolutionise treatments for breast cancer and advance the field of oncology as a whole. Keywords: Inonotus obliquus, Antrodia camphorata, Phellinus linteus 97 1 2 1 1,2*
Presenter’s ID: PB08 Category: Non-Communicable Disease THE EFFECTS OF ECZEFOLIA™ ON THE MIGRATION OF HCT116 COLON CANCER CELLS Nur Sorfeena Sufi Mara Ridhuan, Nur Fariesha Md Hashim, Nurul Akmaryanti Abdullah, Noraina Muhamad Zakuan Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. *Corresponding author email address: [email protected] Colon cancer ranks third as most common cancer globally and its primary cause of death was often metastasis. Therapeutic interventions such as radiotherapy, chemotherapy and surgery utilised in treating colon cancer are receiving drawbacks due to manifestation of toxic side effects, drug resistance and recurrence. Eczefolia™ is an enriched extract of Melicope ptelefolia (MP), commonly known as ‘Tenggek Burung’ in Malaysia, which recently shown to have anticancer effects. It is enriched with 2,4,6-Trihydroxy-3-geranyl acetophenone (tHGA), a compound demonstrating a range of pharmacological properties such as anti-inflammatory, antioxidant, anti-allergic and anti-cancer properties. Hence, the study on of Eczefolia™ may provide some preliminary insights on its anticancer properties, specifically on the inhibition of migration on colon cancer cells. This study aims to determine the cytotoxicity and migratory effects of Eczefolia™ on HCT116 colon cancer cells. Colon cancer cells (HCT116) were exposed to various concentrations of Eczefolia™ (3.9 µg/mL to 1000 µg/mL) to determine the cytotoxicity of the extract using cell viability (MTT) assay. Scratch migration assay was performed to determine the migratory effect of colon cancer cells following Eczefolia™ treatment. Based on the MTT assay result, IC20 and IC50 could not be obtained. The scratch migration assay showed a significant reduction in the migratory rate of HCT116 cells treated with higher concentration of Eczefolia™ (1000 µg/mL ) (p <0.05). Based on the findings, Eczefolia™ did not exert cytotoxic properties at any given concentration, suggesting that its cytotoxic concentration is >1000 µg/mL. The study of migration showed that Eczefolia™ can reduce migratory rate of HCT116 at concentration of 1000 µg/mL, suggesting the potential effects of Eczefolia™ in reducing cancer cell migration, thus invasion. Eczefolia™ does not exhibit cytotoxic properties towards HCT116 cells but shows anti-migratory properties. Therefore, it could be one of the potential candidates to reduce cell invasion and metastasis. Keywords: Eczefolia™, Melicope ptelefolia, HCT116 98 *
Presenter’s ID: PC01 Category: Others EFFECTS OF HCoV-229E ON HUMAN PLATELET AGGREGATION INDUCED BY EPINEPHRINE Nur Tha’iyyah Kamaruzaman , Farah Syazwanie Mohd Zahiruddin , Nadia Nabilah Mohd Marphy , Mohammad Azri Faiq Mohammad , Zetty Nadia Mohd Zain , Zulkefley Othman Department of Biomedical Science, Faculty of Medicine and Health Sciences, University Putra Malaysia, Serdang, Selangor, Malaysia. Department of Medicine, Faculty of Medicine and Health Sciences, Islamic University Science Malaysia, Nilai, Negeri Sembilan, Malaysia. *Corresponding author email address: [email protected], [email protected], [email protected] Human coronaviruses, specifically SARS-CoV-2 that causes COVID-19, are linked to thrombosis, which can result in serious consequences. In COVID-19 patients, the cytokine storm and the interaction of SARS-CoV-2 with ACE-2 receptors lead to persistent inflammation and the development of thrombosis. This study aims to determine the effects of Human Coronavirus 229E (HCoV-229E) on human platelet aggregation induced by epinephrine. HCoV-229E was cultured in MRC-5 cells. Platelet rich plasma (PRP) and platelet poor plasma (PPP) were obtained from healthy individuals (N=3). The PRP then pre-incubated with HCoV-229E (MOI 0.001 and MOI 0.1) for 30 minutes before being added into a 96-well plate containing various concentration of epinephrine (0.001 μM, 0.01 μM, 0.1 μM, 1.0 μM, 10 μM and 100 μM). Modified light transmission aggregometry (LTA) is used to measure aggregation. Data were analyzed by using GraphPad Prism 10.2, The result is expressed as mean ± SEM and an independent t-test was performed to compare each group. In low titre of HCoV-229E, platelet aggregation of control at concentrations 1.0 μM, 10 μM and 100 μM is high (7.17 ± 4.94%; 10.3 ± 7.54%; 16.83 ± 5.9%) as compared to HCoV-229E infected platelet (0.83 ± 2.6%; 7.83 ± 2.85%; 15.67 ± 1.87%). However, in high titre of HCoV-229E, infected platelet showed an increase in platelet aggregation (32.0 ± 23%; 64.0 ± 23%; 68.5 ± 17.5%) as compared to control (1.0 ± 2%; 2 ± 7.5%; 7.5 ± 3.5%). The experiment demonstrated that platelet aggregation is decreased in the presence of low titers of Human Coronavirus 229E (HCoV-229E) when induced by epinephrine. However, when exposed to high concentrations of the virus, platelet aggregation induced by epinephrine is increased. As a conclusion, early coronavirus infection may cause an inhibition of platelet activation but exacerbate procoagulation state when virus infection is high. Keywords: Epinephrine, SARS-Cov-2, aggregation 99 2 1 1* 1 1* 1 2 2*