Poster 1.2

Profiling of Exosomal Circular RNA in Folfox-Resistant

Kha Wai Hon1, Nadiah Abu1*, Nurul-Syakima Ab Mutalib1 and Rahman Jamal1
1UKM Medical Molecular Biology Institute (UMBI), Malaysia

Corresponding author: Dr. Nadiah Abu,


Exosomes are extracellular nanovesicles released by almost all living cells including colorectal cancer cells (CRC) into biological fluids. Exosomes are natural carriers of proteins and nucleic acids that reflects parent cells. Exosomes have been investigated extensively as potential biomarkers and therapeutic targets for various diseases. However, the potential of exosomes as biomarkers to detect Folfox-resistant CRC has not been fully explored and validated. This research aimed to characterise exosomes derived from Folfox-resistant HCT116 CRC cell line as potential targets for further development of biomarkers among CRC patients with chemo-resistance.


Normal CRC cell line HCT116 (HCT116-C) was induced with 5 cycles of drug treatment to develop corresponding Folfox-resistant derivative clones (HCT116-R). HCT116-C and HCT116-R cell lines were compared for cell viability in 5-FU and oxaliplatin, wound healing assay, migration and invasion. Exosomes derived from both cell lines were characterised by transmission electron microscopy, particle size measurement, zeta potential analysis and Western Blot. Total RNA isolated from the exosomes of each cell line was subjected for circular RNA microarray. Selected circular RNAs were validated in clinical samples via RT-PCR.


As compared to normal HCT116-C cell line, HCT116-R cell line had higher IC50 values with 2.5 to 6-fold change. Migration rate of HCT116-R cell line was 19 percent higher than HCT116-C cell line in wound healing assay. Exosomes from both cell lines were morphologically cup-shaped nanovesicles that range from 60 to 370nm in size with zeta potential between -29.1mV and -16.3mV. 105 circular RNAs were upregulated and 34 circular RNAs were downregulated in exosomes derived from HCT116-R against HCT116-C cells.


We developed a stable HCT116-Folfox resistant CRC cells, and the characteristics of exosomes derived from parental and resistant cell lines is in concordance with the literature. We have identified several circular RNAs in exosomes as potential biomarkers to predict Folfox-responsiveness.

Keywords: colorectal cancer, molecular target, FOLFOX, biomarker, Exosomes

Poster 1.3

Efficacy of inhibition of BAD Ser99 phosphorylation by a novel small molecule in cisplatin resistant ovarian cancer

Yanxin Wang1*
1National University of Singapore, Singapore
Corresponding author:  Ms. Yanxin Wang,

Human BAD is a pro-apoptotic Bcl-2 family member, whose apoptotic functions can be inactivated through phosphorylation of specific residues including Ser99. Clinically, BAD phosphorylation has been reported to indicate poor survival and cisplatin resistance in ovarian cancer patients.

NPB, a novel small molecule which specifically inhibits BAD Ser99 phosphorylation, has been developed in our laboratory. Cell function assays performed include Alamar Blue cell viability assay, caspase3/7 assay, PI-Annexin V apoptosis assay, and 3D growth in Matrigel. The CI values are calculated using Chou-Talalay method. The cancer stem cell-like cell population was examined by ALDEFLOUR and sphere formation assays.

The level of BAD Ser99 phosphorylation is negatively correlated with cisplatin sensitivity in a panel of ovarian cancer cell lines and it is increased upon acute cisplatin treatment in ovarian cancer cells. The inhibition of BAD Ser99 phosphorylation by NPB alone increased apoptosis, decreased anchorage-independent growth of both parental and cisplatin resistant ovarian cancer cells, with the combination of NPB and cisplatin showing a synergistic effect. An upstream BAD kinase, AKT has also been reported to mediate cisplatin resistance. Correspondingly, the combination of NPB and AKT inhibitor AZD5363 exhibited strong synergistic effects in both parental and cisplatin resistant cell lines. Both the phosphorylation of BAD and its upstream kinase AKT was increased in this CSC-like population. NPB treatment alone was observed to decrease the CSC-like population, while the combination of AZD5363 and NPB produced a synergistic decrease in the CSC-like population.

NPB, as a novel inhibitor of BAD Ser99 phosphorylation, can potentially be used in combination with cisplatin as a therapy for naïve ovarian cancer patients to increase cisplatin sensitivity. Furthermore, the combination of NPB and AKT inhibitor AZD5363 is a potential therapeutic approach in the treatment of cisplatin resistant ovarian cancer.

Keywords: Bad phosphorylation, cisplatin resistance, ovarian cancer, NPB, Akt inhibitor

Poster 1.4

The Mutagenic Study of Benzimidazole Analogues: Structure-Activity Relationship (SAR)

Nurul Hafizan Azahar1, Siti Soleha Abdullah1, Rozaini Abdullah2, Norizan Ahmat3 and Hasiah Ab Hamid1*
1Department of Biomedical Sciences, Faculty of Medicine and Health sciences, University of Putra Malaysia, Malaysia
2Department of Environmental and Occupational Health, Putra Malaysia University, Malaysia
3School of Chemistry and Environment, University Technology Mara, Malaysia

Corresponding author: Dr. Hasiah Ab Hamid,


Benzimidazole analogues have diverse range of biological activities including anti-ulcer, anti-hypertensive, anti-viral, anti-fungal, anti-inflammatory and anti-cancer. Previous studies revealed some of the analogues can induce mutation despite their biological activities. This study aims to screen for potential mutagenic activities of novel benzimidazole analogues using Ames test and study their structure-activity relationship (SAR).


Ames test was carried out on two strains of Salmonella typhimurium; TA98 and TA100 that carry different mutations in their various genes in histidine operon, which protocols complied with OECD 471 Guideline for Testing of Chemicals. The preliminary tests including growth curve and genetic analysis were carried out prior to Ames test in order to determine the optimum incubation hour and genotypes of the bacterial strains respectively. The bacterial growth curve was carried out by incubating the bacterial suspension for 52 hours. The optical density of the bacteria was recorded for every two hours at 660nm wavelength and the graph of optical density versus time was plotted. The genotypes of the bacterial strains used were determined by genetic analysis which includes tests on dependency on histidine and/or biotin, presence of rfa mutation, urvB deletion as well as pKM101 plasmid. All data were analysed using Two-Way ANOVA, followed by post-hoc Tukey test. The SAR was analysed based on the results of Ames test.


The optimum incubation hour for both strains was 14 hours. Both bacterial strains show dependency on histidine with presence of rfa mutation, urvB deletion and pKM101 plasmid. Besides, all analogues tested; NN-1-5, NN-1-7, NN-1-9 and NN-1-18 show no mutagenic activity.


There was no significant difference in the mutagenic activity among all analogues despite their difference in the number and position of hydroxyl groups.

Keywords: benzimidazole, Ames test, Salmonella typhimurium, Structure-activity relationship (SAR), MUTAGENIC

Poster 1.5

Drug-herb interaction between methanolic and aqueous extracts of Clinacanthus nutans and cisplatin on breast cancer cell line MCF-7

Nurliana Abd Mutalib1, Normala Abd Latip1*, Farah Athirah Mohd Nawawi1 and Nurul Anieza Osman1

1Universiti Teknologi MARA Puncak Alam, Malaysia
Corresponding author:  Dr. NORMALA ABD LATIP,

Drug-herb interactions may affect drug therapy in several ways leading to beneficial or even harmful consequences. Significant numbers of patients all around the world consume natural products as complementary to conventional medicine in the battle against various diseases including cancer. Clinacanthus nutans locally known as Sabah snake grass is popular for its anti-cancer properties and taken as prevention as well as treatment for cancer. It is crucial to evaluate potential antagonistic, additive or synergistic interactions that may result from co-treatment of this plant in chemotherapy. In this study, after preliminary cytotoxicity test of methanolic (MCN) and aqueous (WCN) extracts of C. nutans on a panel of cell lines was conducted. MTT assay was used to test single treatment as well as the combination of extracts and cisplatin (CP) on MCF-7, breast cancer cell line. From the data, combination indexes were calculated and isobolograms were constructed. CP, MCN and WCN decreased the survival of breast cancer cells with IC50 of 43.59 µM, 378.44 µg/mL and 910.92 µg/mL respectively. Combination of WCN and CP exhibited strong antagonism. Most combination of MCN and CP also generated significant antagonistic effect except for combination of CP with high concentration of MCN which is additive (CI=1). MCN has greater cytotoxic affect against MCF-7 compared to WCN and both showed strong antagonism when used in combination with CP.

Keywords: Clinacanthus nutans, Combination index, Isobologram, MCF-7, Cancer

Poster 1.7

Farmed Edible Bird’s Nest in Malaysia: Proximate Analysis, Safety Profile, Epidermal Growth Factor Content and Effect on Cancer Cells

Sin N. Tan1*, Careena Shobana1, Sani Dahiru1, Chee W. Lim1, Ideris Aini1, Stanslas Johnson1 and Thiam S. Christopher Lim1
1Putra Malaysia University, Malaysia
Corresponding author: Dr. Sin N. Tan,


Edible bird’s nest (EBN), a solidified swiftlet’s saliva, is the most expensive animal products consume by human worldwide. EBN exhibits multiple medicinal properties and contains many bioactive compounds such as sialic acid, amino acids, fatty acids, minerals and antioxidants. In recent years, many safety issues which included high nitrate and nitrite contents, presence of heavy metal, adulteration, fungal infection and cancer cell stimulation were found to be associated with EBN. Hence, this study was carried out to determine the proximate analysis, safety profile, epidermal growth factor content and its effect on cancer cells of Malaysia EBN.


Proximate analysis and safety profile were performed by using official AOCA methods according to the Malaysia Standard MS 2509:2012. Coupled Plasma Optical Emission Spectrophotometer was adopted for heavy metals analysis. Enzyme-linked immunosorbent assay was used for quantification of EGF content. Microculture tetrazolium assay was utilized for assessment of growth stimulation by different concentration of human epidermal growth factor (hEGF) in comparison to EBN in 4 different cancer cell lines: MCF-7 (human breast adenocarcinoma cells), Caco-2 (human epithelial colorectal adenocarcinoma cells), HCT116 (human colorectal carcinoma), and A549 (human alveolar adenocarcinoma cells) for 24 hours, 48 hours, and 72 hours.


Our results showed a consistency of high protein (53.03% – 56.37%) and carbohydrate (27.97%-31.68%) with acceptable level of moisture (10.8-14.04%) and ash (2.22%-3.38%). Besides, a good safety profile was obtained with low nitrite and nitrate level, undetectable heavy metals and no significant growth for pathogenic microorganism. The present study found above tolerance level of yeast and moulds in EBN. Haze contaminated EBN exhibited no significant differences in terms of nutritional, heavy metal and microbiology profiles. EFG (30.7pg/ml and 74.5pg/ml) were detected in crude EBN01 and EBN02, respectively but not in all digested EBN samples and even post 10 times concentrated EBN. Cancer cell growths were significantly increased after treatment with hEGF. No significant growth was observed after treatment with EBN.


EBN in Malaysia is free from heavy metals, within tolerance level of nitrate and nitrite, and also microbiology profile. Furthermore, in vitro study indicated that EBN is not associated with cancer cell stimulation.

Keywords: Edible bird nest, nutritional, Safety, Epidermal Growth Factor, Cancer

Poster 1.8

The antioxidant and cytotoxic properties of Morinda citrifolia and Morinda elliptica leaves extracts

Nurulaidah Esa1 and Latifah Saiful Yazan1*
1Department of biomedical science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia

Corresponding author: Prof. Latifah Saiful Yazan,


Cancer is one of the top ten causes of hospitalisation and death in Malaysian hospitals. Standard treatments for cancer patients include surgery, chemotherapy and radiotherapy, but with several adverse effects. Hence, researchers have shifted their focus on natural resources as potential treatment. Morinda citrifolia and M. elliptica of the Rubiaceae family have been traditionally used to treat diabetes, gastric, ulcer, inflammation and cancer. The plants were found to exert these promising therapeutic effects given their antioxidant properties. This study investigated the antioxidant and cytotoxic properties of M. citrifolia and M. elliptica leaves extracts.


Morinda ethanol extracts were prepared by immersing the leaves for one day in 50, 70 or 100% ethanol (1:10, w/v) at room temperature while the aqueous extracts were boiled in distilled water for 0, 5, 15, 30 and 60 minutes (1:100, w/v). The extracts were subjected to antioxidant assays – total phenolic content (TPC), 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay, 2,2-azinobis-3-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and ferric ion reducing antioxidant power (FRAP). The cytotoxicity of the extracts on human colorectal adenocarcinoma cell line (HT-29) were determined using MTT assay.


The M. elliptica aqueous extract has the highest antioxidant activities among all the extracts. M. elliptica, boiled for 30 minutes has the highest TPC (75.98+4.53 mg GAE/g extract) and while at 0 minute, it has the highest FRAP (176.33+4.4 TEAC/g extract). M. elliptica has the highest DPPH (18.22+0.62 mg TEAC/g extract) and ABTS (219.84+2.93 mg TEAC/g extract) radical scavenging activity when boiled for 5 minutes. The M. elliptica extract in 100% ethanol was the most cytotoxic among all the extracts with IC50 of 105.5 + 1.0 µg/mL at 72 hours.


The M. elliptica leaves extracts have better antioxidant and cytotoxic properties when compared to the M. citrifolia leaves extracts.

Keywords: Morinda citrifolia, Morinda elliptica, antioxidant, cytotoxic , HT-29

Poster 1.9

An ethnobotanical survey and in vitro cytotoxicity study of medicinal plants traditionally used in the treatment of cancers in Ugbine, Nigeria

Ogochukwu N. Nwaefulu1, Josephine O. Owolabi2, Sreenivasa R. Sagineedu3, Lim C. Woei1, Nur K. Kassim1 and Johnson Stanslas1*

1Putra Malaysia University, Malaysia
2University of Benin, Nigeria
3International Medical University, Malaysia

Corresponding author: Prof. Johnson Stanslas,


Herbalists in Ugbine, Nigeria are known to use herbs in treatment of cancers amongst other diseases. Therefore, this research was designed to study the medicinal plants used in the treatment of cancers amongst traditional healers in the community and herb sellers, and to validate this claim scientifically.


A cross–sectional survey was conducted between August and September 2017 using a validated structured questionnaire. Plant specimens indicated in the recipes were collected, sent to Forestry Research Institute of Nigeria (FRIN) for identification and authentication, thereafter used for cytotoxicity study. A specimen with voucher number FHI – 111105 was deposited. Dried/powdered leaves 45 g of Petiveria alliacea (PA) was extracted with 900 ml of dichloromethane (DCM), methanol (MeOH) and DCM:MeOH (1:1) by cold maceration for 72h. The solvent extract was filtered and concentrated using rotary evaporator, subsequently was dried in an oven at 400C for 3 days. 3-(4,5-Dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was performed to assess in vitro growth inhibition of extracts against cancer cell lines (HCT-116 (colon), PC-3 (prostate), MCF-7 (breast), PANC-1 (pancreas). Cells were treated with the extracts (0.01 to 100 µg/ml) for 96 h at 5% CO2 and 37oC. An extract was regarded to be active if it had a mean GI50 < 100 µg/ml in any of the four cell lines. Among the active extracts, GI50 < 10 µg /ml in at least one tumour cell line was classified as strong growth inhibitory effect, whereas extracts with GI50 in the range of 10-100 µg /ml were considered to have moderate growth inhibitory effect. Extracts with GI50 > 100 µg/ml were considered to have no activity.


A total of 11 respondents were interviewed. They were mainly herb sellers (64%) and the rest were traditional healers. They were predominantly females (90.9%). Majority (63.6%) of them aged between 61-90 years, while 36.4% were between 31 – 60 years old. The level of illiteracy amongst them is high (63.6%). The tribe that predominates amongst the respondents was Yoruba (45.5%), followed by Bini (36.4), Calabar (9.1%) and others (9.1%). Majority (54.5%) of them practised African traditional religion, followed by Islam (27.3%) and Christianity (18.2%). Although majority (81.8%) of the respondents have knowledge of the disease called cancer, the treatment of this disease with herbs was only practised by 9.1% of them. The in vitro cytotoxicity test showed moderate growth inhibitory effect of DCM:MeOH extract against HCT-116 (GI50 55 ± 10.4 µg/ml), PC-3 (GI50 65 ± 5.0 µg/ml) but failed to show activity against PANC-I and MCF-7 cells. The DCM extract also showed moderate activity against PC-3 (GI50 45 ± 5.0 µg/ml). However, the MeOH extract was inactive.


PA extracts produced varying degrees of cytotoxic activities against cancer cell lines. This observation therefore supports the potential of this plant as a source of new anticancer agents. Further studies are required to identify the active constituents found in this plant.

Keywords: Petiveria alliacea, Cell viability, cell growth, Cancer, MTT assay

Poster 1.11

Targeting ALK for cancer therapy: high throughput screening assays

Mohamad Fairus Abdul Kadir1, Pratheev Alagapan1, Yang Jin Lim1, Wai Mun Kong1, Dharvind Balan1, Mathura Subangari Dorasamy1, Wesley Roy Balasubramanian2, Sivapriya Marappan2, Umarani Subramaniam1*, Kavitha Nellore2 and Thomas Anthony2
1Aurigene Discovery Technologies Sdn Bhd, Malaysia
2Aurigene Discovery Technologies (India), India

Corresponding author: Dr. Umarani Subramaniam,


Drug discovery is a multi-step, interdisciplinary process which involves target identification, lead optimization, pre-clinical and clinical studies prior to first human exposure. Thousands of chemical compounds designed to interact with/block a specified target are synthesized and isolated at the early stage of a drug development. Subsequently, combinatorial chemistry and assay development are involved to screen selected compounds in a high throughput screening platform. Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase which plays a critical role in proliferation, survival, and differentiation of normal cells. However, continuous activation of ALK signaling results from overexpression and mutation has emerged as a novel tumorigenic player in several human cancers. Crizotinib is the first ALK inhibitor approved by Food and Drug Administration (FDA) followed by Ceritinib and Alectinib. Nevertheless, the development of resistance towards Crizotinib treatment due to the mutations in ALK (L1196M, C1156Y, and F1174L) triggered the development of alternative pharmacologic agents that inhibit the mutant forms as well. Therefore, in vitro ALK assays using Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) and In-Cell Western (ICW) technology were developed to identify potential compounds that could inhibit the activity of both the ALK wild-type and its mutant form.


In TR-FRET assay, enzyme was added to a reaction mix containing buffer, substrate and ATP in a 384-well plate. For ICW assay, Karpas-299 cells were seeded in the microtiter plates followed by fixation and permeabilization. The inhibitory potency of the standard (NVP-TAE684, Crizotinib and Staurosporine) and test compounds was detected by the phospho-ALK with state-specific antibodies and conjugated secondary antibodies. The intensity of light emission is proportional to the level of substrate/ALK phosphorylation and the percentage inhibition of the compounds was then determined.


In general, the standard and test compounds were more potent for WT ALK as compared to the L1996M mutant. The biochemical potency for WT and mutant ALKs correlated well with the cell potency in Karpas-299 cells.


The TR-FRET and ICW assays established with standard inhibitors can be used to screen compounds for their potential to inhibit ALK activity. The assays are simple, cost effective and ideal for high throughput screening to develop potential ALK inhibitors for cancer therapy.

Keywords: TR-FRET, In-cell Western, kinase, Drug Discovery, Cancer

Poster 1.12

Neural networks approach to optimization of aerosolized nanocolloidal carrier system containing quercetin for pulmonary delivery of lung cancer

Mohd Basyaruddin Abdul Rahman1*, Noor Hafizah Arbain1, Norazlinaliza Salim1 and Wong Tin Hui2
1Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, Malaysia
2Non-Destructive Biomedical and Pharmaceutical Research Centre, Integrative Pharmacogenomics Institute, Universiti Teknologi MARA, Malaysia

Corresponding author: Prof. Mohd Basyaruddin Abdul Rahman,


Globally, lung cancer has become the most common type of cancer cases. Quercetin (QT) has been extensively investigated for its pharmacological effects on lung cancer. However, clinical applications of QT are limited due to poor solubility and low stability in aqueous medium. Hence, this study focused on the development of nanocolloidal carrier system to enhance the solubility of QT by pulmonary administration.


The artificial neural networks (ANNs) was carried out for development of a predictive model of aerosolized quercetin (QT) nanoemulsions to achieve minimum volume median diameter for targetting lung cancer by pulmonary delivery. The effects of the amount of the mixture of palm oil esters:ricinoleic acid (1.5–4.5 wt.%), lecithin (1.5–2.5 wt.%), Tween 80 (0.5–1 wt.%), glycerol (1.5–3 wt.%), and water (88–94.95 wt.%) on the volume median diameter were considered as inputs to the network. The volume median diameter of the samples with various compositions was measured as an output. The incremental back propagation (IBP), batch back propagation (BBP), quick propagation (QP), and genetic algorithm (GA) were used in the network.


It was found that the optimal algorithm and topology were the genetic algorithm (GA) and the configuration with 5 inputs, 14 hidden, and 1 output nodes, respectively. The model obtained indicated the high-quality performance of the neural network and its capability to identify the critical composition factors for the nanoemulsion. The formulation containing quercetin was then successfully prepared using optimum composition and volume median diameter of 4.266 μm was obtained. The QT-loaded nanoemulsion induced cytotoxic response with an IC50 value of 300 μg/mL against A549 lung cancer cells at 48 h.


These results suggest that the nanocolloidal formulation containing QT could be successful carrier system for pulmonary drug delivery application.

Keywords: Aerosols, Quercetin, Nanoemulsions, Pulmonary delivery, Arti cial neural network

Poster 1.13

Semisynthetic Derivatives of Andrographolide Inhibit Growth of Pancreatic Cancer Cells via Induction of G1 Phase Cell Cycle Arrest and Apoptosis

Sopna Devi Revindran1 and Johnson Stanslas1*
1Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia

Corresponding author: Prof. Johnson Stanslas,


Pancreatic ductal adenocarcinoma, is currently the fourth leading cause of cancer death globally due to poor prognosis and late diagnosis. Mutation of Kirsten-Ras (K-Ras) oncogene is prevalent in pancreatic cancer patient as more than 90% of patients inherit it. Andrographolide (AGP) and its benzylidene derivatives were found to bind to K-Ras transient pockets resulting in inhibition of K-Ras-MAPK oncogenic signaling. In the current investigation, growth inhibition, cell cycle arrest and apoptosis inducing potential of semisynthetic benzylidene derivatives of AGP, namely 3,19-(2-bromobenzylidene) andrographolide (SRJ09), 3,19-(3-chloro-4-fluorobenzylidene) andrographolide (SRJ23) and 3,19-(2-bromobenzylidene)-14-acetylandrographolide (SRS07) were evaluated in Capan-2 (K-Ras G12V mutation) and PANC-1 (K-Ras G12D mutation) pancreatic cancer cell lines.


In vitro cytotoxic activity of AGP derivatives was assessed at 96 hours using MTT cell viability assay. Non-treated and treated cells were fixed in 70% ethanol and stained with propidium iodide before being subjected to quantification of cellular DNA content at different cell cycle stages by BD FACSCANTO II. Annexin V in combination with 7-Aminoactinomycin D (7-AAD) dye were used to stain cells, followed by analysis using Muse Cell Analyser that utilise fluorescent detection to detect phosphatidylserine (PS) on the surface of apoptotic cells.


All three AGP derivatives were equally potent in Capan-2 and PANC-1 cells with IC50 values ranging from 3.2 µM to 4.9 µM. Cell cycle analysis showed that AGP analogue induced cell cycle arrest in G1 phase. Apoptosis assay revealed a decrease in viable cells, with concomitant increase in early and late apoptotic cells within 48 hours of exposure to AGP analogue.


The benzylidene derivatives of AGP inhibited growth of pancreatic cancer cells by inducing G1 phase cell cycle arrest and apoptosis.

Keywords: Cell Cycle, Apoptosis, MTT assay, Pancreatic Cancer, andrographolide

Poster 1.14

Assessment of Growth Inhibitory, Cell Cycle Arrest, and Apoptosis Inducing Potential of RAS-MAPK Inhibitors in Gemcitabine-Sensitive Pancreatic Cancer Cells

Nityaa Selvarajoo1 and Johnson Stanslas1*
1Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia

Corresponding author: Prof. Johnson Stanslas,


Pancreatic cancer is an aggressive disease. Although its occurrence rate remains low in Malaysia, it is the third leading cause of cancer-related death in the United States of America because of limited treatment options. Gemcitabine is a standard chemotherapy for pancreatic cancer. Most pancreatic cancers harbour K-Ras mutations that lead to constitutive activation of mitogen-activated protein kinase (MAPK) signalling pathway. The aim of this study was to determine the cell cycle and apoptosis inducing potential of RAS-MAPK inhibitors such as B-Raf kinase inhibitor (vemurafenib), a Mek1/2 kinase inhibitor (selumetinib) and Erk1/2 kinase inhibitor (ulixertinib) in gemcitabine-sensitive pancreatic cancer cell lines.


3-(4,5-Dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used in assessing in vitro growth inhibition of RAS-MAPK inhibitors against pancreatic cancer cell lines (Panc-1 – mutated KRAS, wild-type BRAF, MEK, and ERK; Colo-357 – mutated KRAS and BRAF, wild-type MEK and ERK; Capan-2 – mutated KRAS, wild-type BRAF, MEK, and ERK); Miapaca-2 – mutated KRAS, wild-type BRAF, MEK, and ERK); BxPC-3- wild-type KRAS, BRAF, MEK and ERK). Cells were treated with 0.1-100 µM of RAS-MAPK inhibitors alone or with gemcitabine and the 50% cell viability- inhibition values were obtained from dose-response curves at 96 hours time-point. Subsequently, flow cytometry was utilised to analyse cell cycle arrest, while Muse® Annexin V & Dead Cell Kit (Merck) was performed to determine apoptosis.


Ulixertinib exhibited strongest inhibitory effect on Capan-2 with an IC50 value of 0.64 µM, while it showed moderate activity (3.2 µM – 7.25 µM) in other pancreatic cancer cell lines. Vemurafenib and selumetinib were less active against all cell lines. Ulixertinib at 1 µM induced G1 cell cycle arrest in Capan-2. In apoptosis assay, treatment with 1 µM induced 29% and 20.3% of early and late apoptotic cells population, respectively.


The results suggested that Ulixertinib inhibited growth of Capan-2 cells by inducing G1 arrest and apoptosis. It is presented itself as the most active RAS-MAPK inhibitor against the molecularly selected pancreatic cancer cell lines.

Keywords: RAS-MAPK inhibitors, gemcitabine, Apoptosis, Pancreatic Cancer, Cell Cycle

Poster 1.15

Development of novel inorganic strontium nanoparticles for delivery of pDNA and siRNA to breast cancer cell

Athirah Bakhtiar1*, Md Ezharul H. Chowdhury2, Iekhsan Othman3 and Anuar Zaini3

1Faculty of Pharmacy, Mahsa University, Malaysia
2Monash University Malaysia, Malaysia
3Monash University, Australia

Corresponding author: Dr. Athirah Bakhtiar,


Recent studies have been focused on developing smart nanoparticles for efficient delivery of transgenes and siRNAs into cancerous cells of an animal model through active and passive targeting. Precipitation reaction is one of the facile and convenient ways to generate inorganic strontium nanoparticles, in which an insoluble salt is formed after mixing two water-soluble salts. Advantages in comparison with other methods include requiring only simple equipment, ability to prepare and control particle size and composition in near ambient temperature and pressure. The approach would pave a new way for facile, low toxicity and large-scale synthesis of inorganic nanoparticles with well-controlled dimensions and properties.


The study involved the formation of 5 different types of inorganic strontium nanocrystals via precipitation reactions, with various observational studies involving pH, temperature, salt concentration and time of incubation. Strontium particle sizes were determined using zetasizer and scanning electron microscope prior to binding studies. The nanoparticles were then subjected to various cellular studies including cellular uptake and cytotoxicity studies through incorporation of p53 and MAPK siRNA, forming gene-nanoparticle complexes onto human and mice mammary carcinoma cells, MCF-7 and 4T1 cells, respectively. Unloaded strontium nanoparticles formed were also observed for toxicity effect against both cells.


Studies showed that strontium fluoride and strontium carbonate were smaller in size in comparison to other strontium salt particles, averaging in 100-300nm in diameter. Additionally, cellular studies revealed that gene-loaded strontium sulfite and strontium fluoride were responsible for greater cytotoxicity effect, in comparison naked gene treatment. Low toxicity effect was seen with unloaded strontium nanoparticles. Studies were compared against established inorganic carbonate apatite nanoparticles.


Nano-sized strontium particles showed great potential to be applied for in vivo studies, ideally seen with strontium sulfite and strontium fluoride, via the gene delivery system in the cellular level.

Keywords:  breast cancer, pDNA delivery, siRNA, Strontium, Nanoparticles

Poster 1.16

Tongue Cancer in Elderly Patients: Case Report of A 69 Year Old Patient With Dental Pain Complain

Mahibub M. Kanakal1*, Shah Kamal Khan Jamal Din1, Amin Malik Shah Abdul Majid2, 3, KHAIRULANWAR HUSAIN1 and Mohammed Amir Shah Abdul Aziz4

1Quest International University Perak, Malaysia
2Universiti Sains Islam Malaysia, Malaysia
3Department of Pharmacology, School of Pharmaceutical Sciences, University of Science Malaysia, Malaysia
4Penang General Hospital, Malaysia

Corresponding author:  Dr. Mahibub M. Kanakal, Quest International University Perak, Ipoh, Malaysia,

 Less than 15-25% of those who visit a dentist regularly report having had an oral cancer screening. Only approximately 2% of patients are diagnosed with oral cancer before the age of 35. Current study reports the case of a 69-year-old man who visited his dentist with initial complains of chronic dental pain for 9 months, leading to diagnosis of squamous cell carcinoma of the tongue. Patient was unable to chew his food on both sides of his jaws. He habitually chews more on one side due to occasional pain referred to his Left Temporal Mandibular Joint (LTMJ) for a period of 9 months. Patient received a routine dental treatment after full examination. A dental bridge was constructed on both sides of his molar region as 3 and 4 unit porcelain bridge and discharged. Six months later patient had returned to clinic complaining of discomfort on the right lateral side of his tongue which shifted to the right side of the tongue margin which became sore and painful. During close examination a whitish rough area on his tongue surface with ulceration was noticeable. Biopsy was done under local anaesthetic using Scandonest and Mepivicaine HCl. Biopsy results confirmed a diagnosis of ventral lateral side of right tongue Squamous Cell carcinoma. Squamous cell carcinoma is possibly under diagnosed in the elderly patients and patients with dental pain complains especially relating to the tongue area should be closely assessed and monitored.

Keywords: squamous cell cancer, case report, elderly patient, Dental pain, Cancer


Poster 2.2

Differentiation of mitogen-activated protein kinase-dependent deregulated myokine production in inflamed skeletal muscle cells

Divya Malathy Ravinath1 and Ren-In You1*
1Tzu Chi University, Taiwan

Corresponding author: Dr. Ren-In You,


Skeletal muscles, when encountered with damage due to exercise, stress or myotrauma triggers regeneration process which includes the destructive/inflammatory phase, repair phase and remodeling phase. During the inflammatory phase, some myokines are produced which are involved in anti-inflammatory and immuno-modulatory roles. Deregulated expression of myokines due to abnormal inflammatory response serves as an important cause of inflammatory myopathies or severe muscle weakness.


In order to understand the underlying mechanism of inflammatory myopathies, the model system used was C2C12 myoblast cells cultured in low serum condition. Many inflammatory cytokines were used to stimulate the cells, thereby mimicking the inflammatory response. The morphology and fusion of myoblasts into multi-nuclear myotubes were demonstrated using Hoechst 33342 under fluorescent microscope. In addition, kinase inhibitors, SB203580, PD98059 and U0126 were used to investigate the involvement of mitogen-activated protein kinase (MAPK) signaling pathway during muscle inflammation condition. The gene expression variation of myokines was analyzed using qPCR. Finally western blot was performed to analyze the corresponding variation in protein expression of downstream kinases of MAPK pathway.


We found that, certain pro-inflammatory cytokines triggers the downstream MAPK signaling in C2C12 skeletal muscle cells. The inhibition of MAPK during inflammation stimulatory condition led to a significant increase of the myokines but inhibited the differentiation of myoblasts. The downstream of MAPK were also found to be selectively expressed in different time frames.


The MAPK pathway plays an important role in the regulation of inflamed muscle cells. In this study, the role of mitogen-activated protein kinase kinase (MEK) in altering the myokine expression during inflammatory conditions will be depicted.

Keywords: Inflammatory myopathy, Myokines, mitogen activate protein kinase, skeletal muscle, signaling

Poster 2.3

Celastrol alleviates gamma irradiation-induced damage by modulating diverse inflammatory mediators

Hong Wang1* and Gautam Sethi1*
1National University of Singapore, Singapore
Corresponding authors: Dr. Hong Wang, and Prof. Gautam Sethi

The present study was aimed to explore the possible radioprotective effects of celastrol and relevant molecular mechanisms in an in vitro cell and an in vivo mouse model exposed to gamma radiation. Human keratinocyte HaCaT and BJ skin fibroblast cells were exposed to gamma radiation of 20Gy, followed by treatment with celastrol for 24h. Cell viability, reactive oxygen species (ROS), nitric oxide (NO), and glutathione (GSH) production, lipid peroxidation, DNA damage, inflammatory cytokine levels, and NF-κB pathway activation were examined. The survival rate, levels of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) in blood, and p65 and phospho-p65 expression were also evaluated in mice after exposure to gamma radiation and celastrol treatment. The gamma irradiation of HaCaT cells induced decreased cell viability, but treatment with celastrol significantly blocked this cytotoxicity. Gamma irradiation also increased free radical production, e.g. ROS and NO, decreased the level of GSH, and enhanced oxidative DNA damage and lipid peroxidation in cells, which were effectively reversed by celastrol treatment. Moreover, inflammatory responses induced by gamma irradiation, as demonstrated by increased levels of IL-6, TNF-α, and IL-1β, were also blocked by celastrol. The increased activity of NF-κB DNA binding following gamma radiation was significantly attenuated after celastrol treatment. In the irradiated mice, treatment with celastrol significantly improved overall survival rate, reduced the excessive inflammatory responses, and decreased NF-κB activity. As an NF-κB pathway blocker and antioxidant, celastrol may represent a promising pharmacological agent with protective effects against gamma irradiation-induced injury.

Keywords: Celastrol, gamma irradiation, Inflammation, ROS, NF-κB

Poster 2.4

In vitro antipsoriatic activity and mechanism of action of semisynthetic andrographolide compounds

Audrey C. Yong1*, Chai Sia Lim1, Ahmad Zubir Jangsi1, Hon Liong Soo2, Ibrahim Sulaiman2, 3, Yuan Han Teh2, Chee-Woei Lim2 and Johnson Stanslas2
1Faculty of Pharmacy, Mahsa University, Malaysia
2Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia
3Bayero University Kano, Nigeria

Corresponding author: Dr. Audrey C. Yong,


Psoriasis is a chronic disease that is characterised by epidermal keratinocytes hyperproliferation, abnormal differentiation and inflammatory infiltration. Andrographolide (AGP) and 14-deoxy-11,12-didehydroandrographolide (DDAG) were shown to have anti-inflammatory effects. These natural diterpenoid lactones were used as a template for semi-synthesis of a series of new compounds. This research aimed to screen the compounds for in vitro antipsoriatic activity, to study the anti-inflammatory potential and mechanism as well as to analyse the mode of cell death induced by the selected antipsoriatic compounds.


Natural and three semi-synthesised andrographolide derivatives (AGP, DDAG, SRS49, SRS121 and SRS140) were evaluated for antipsoriatic activity in vitro against HaCaT (keratinocytes) cell line. The safety profile was investigated using RAW 264.7 mouse macrophage and Vero monkey kidney normal cell lines. Both assays were evaluated by measuring the formazan absorbance at 550 nm using methylthiazol tetrazolium (MTT) assay. Cell death analysis was performed on selected compounds against HaCaT using the MUSE cell analyser. Griess assay and Western blot were used to study the anti-inflammatory activity and mechanism (respectively) of SRS49 against HaCaT cell line.


SRS49 and SRS140 exerted antiproliferative activity against HaCaT cell line with IC50 value of 57.73 µM and 55.76 µM, respectively. However, SRS140 exhibited moderate cytotoxicity against RAW 264.7 and strong cytotoxicity against Vero. SRS49 compound displayed selective inhibitory properties as it has no inhibitory activity against RAW 264.7 and Vero normal cell lines. Antiproliferative activity of SRS49 against HaCaT could be attributed to apoptosis. SRS49 showed better inhibition (p<0.05) against nitric oxide production when compared to AGP and DDAG post-induction with TNF-α. It is highly probable that the NF-κB transcription factor is also involved in the inflammation pathway.


SRS49 possess in vitro antipsoriatic and anti-inflammatory properties with safer profile. Apoptosis is one of the mechanisms underlying inhibition of HaCaT cell growth by SRS49. This makes SRS49 a suitable candidate that can be developed as an antipsoriatic agent.

Keywords: Psoriasis, Andrographolide (AGP), 14-deoxy-11, 12-didehydroandrographolide , HaCat (Human Keratinocyte), NF-ĸB

Poster 2.5

Phytochemical screening and evaluation of anti-inflammatory of activity of Gardenia carinata leaves

Velaga V. Appalaraju1*, Nahlah E. Ismail2 and senthil Adimoolam2
1Mahsa University, Malaysia
2Faculty of Pharmacy, Mahsa University, Malaysia

Corresponding author: Dr. Velaga V. Appalaraju,


Gardenia carinata plant has wide range of therapeutic uses such as anti-HIV, cytotoxic, antimicrobial and mosquito larvicidal activities. The purpose of this study is to determine phytochemical screening and anti-inflammatory activity of different organic extracts of G. carinata leaves.


The leaf extracts were prepared using different organic solvents in the order of increasing polarity chloroform, methanol and distilled water by using soxhlet extraction method. The extract was characterized by Shinoda test, Salkowski test, Ferric chloride test, Folin-Ciocalteau test and foam test. The anti-inflammatory activity of the entire G. carinata leaf extracts were evaluated at a dose level of 100 mg/kg (bw), orally by using Carrageenan-induced paw edema method. Indomethacin 10 mg/kg (bw) was used as positive control. Paw thickness of rats were monitored every hour post edema induction for 5 hours and the percentage of inhibited edema were calculated.


The inhibitory percentage exhibited by Gardenia carinata water extract (GCWE) was 32.66%, G. carinata methanol extract (GCME) was 27.98% and G. carinata chloroform extract (GCCE) 24.41% respectively. All these groups are significantly differenced in inhibition when compared with Indomethacin inhibition percentage 39.33% at the fifth hour (p < 0.05). Extracts of G. carinata leaves revealed the presence of flavonoids, terpenoids, tannins, phenolic compounds and saponins.


Gardenia carinata leaves showed the anti-inflammatory activity in carrageenan-induced rat paw edema model.

Keywords: Gardenia carinata, anti-inflammatory activity, Albino rats, Intraplantar administration, Carrageenan-induced paw edema model

Poster 2.6

Anti-Inflammatory Activity of Topical Polyherbal Formulation Containing Pereskia bleo and Camellia sinensis Leaves Extract

Javed Qureshi1*, sheryar Afzal1 and Lee C. Kin1
1Faculty of Pharmacy, Mahsa University, Malaysia

Corresponding author: Dr. Javed Qureshi,


Non-steroid anti-inflammatory drugs (NSAIDS) are the most prescribed drugs for the treatment of inflammatory diseases. Furthermore, prolonged use of NSAIDS is associated with severe side effects particularly on gastric mucosa, renal and cardiovascular system. Therefore, searching for new drug candidates in the treatment of chronic inflammation has great importance. Alternative to these drugs are traditional medicines and natural products, which offer a great hope in the identification of bioactive lead compounds and their development into drugs for treating inflammatory diseases. With this regard, the present study was aimed to formulate and evaluate a novel polyherbal ointment containing leaves extract of Pereskia bleo (PB) and Camellia sinensis (CS) for their anti-inflammatory activities.


Fresh air dried leaves of PB and CS were extracted with dichloromethane using Soxhlet apparatus and subjected to phytochemical analysis. Commercially available simple ointment BP (Vaseline®) was used as a base to prepare eight polyherbal ointment formulations containing different concentrations of PB (5% and 10% w/w) and CS (1% and 2% w/w) extracts. Prepared formulations (F1 to F8) were evaluated for physical parameters along with anti-inflammatory activity by using model carrageenan induce paw edema in rats using Diclofenac gel as positive control.


The pH of all formulations ranged between 5.50 to 6.25 which were within acceptable range of pH for topical application. The paw thickness for each group was measured every hour for a total studied period of five hours. The formulation F8 has equivalently reduced the inflammation induced by carrageenan as compared to standard drug Diclofenac. The F8 reduced the inflammation by 36.79% compared to the control treated group (39.5%).


Based on the above observations and results, formulation F8 was found to be a promising anti-inflammatory formulation and further extensive in-vitro and in-vivo studies are warranted to evaluate its biological potency and which may be useful for further clinical applications.

Keywords: Pereskia Bleo, Leaves extract, Antiinflammatory activity, herbal formu

Poster 2.7

Interaction of Palm Kernel Oil Esters with Ibuprofen in Nanoemulsion Drug by Molecular Dynamics Simulation

Nur Hana Faujan1*, Mohd Basyaruddin Abdul Rahman1, Mahiran Basri1 and Roghayeh Abedi Karjiban1
1Department of Chemistry, Faculty of Science, Putra Malaysia University, Malaysia

Corresponding author: Dr. Nur Hana Faujan,

 Palm kernel oil esters have been used to encapsulate lipophilic ingredient in various products such as foods, cosmetics, pharmaceutical, cosmeceuticals and etc. These esters have better physicochemical properties than palm kernel oil, which display non-toxic, non-irritation, low viscosity and less greasy to the skin. Palm kernel oil esters have been applied in the oil phase of nanoemulsion due to its good solubilizing capacity and excellent skin permeation. The hydrophobicity of these molecules could control the self-assembly process and the stability of palm kernel oil esters-based nanoemulsion containing ibuprofen. Relatively, there is limited understanding on the influence of palm kernel oil esters and its interaction with ibuprofen. Here, we focused on the interactions between palm kernel oil esters and ibuprofen in colloidal delivery systems. The palm kernel oil esters-based nanoemulsion system was simulated with 27 molecules of ibuprofen in SPC water for 50 ns. Molecular dynamics simulation was utilized at all-atomic level using GROMACS version 4.5.5. The interatomic interaction between these esters and ibuprofen were analyzed by the radial distribution function, bond interaction and interatomic distance. Oleyl stearate with C18 structure produced strong interaction with ibuprofen due its longest chain in palm kernel oil esters. This allows a higher possibility to interact with ibuprofen molecules. Simulation results provide better understanding on the interaction mechanism between palm kernel oil esters with ibuprofen molecules in nanoemulsion model for drug delivery application.

Keywords: Palm kernel oil esters, Ibuprofen, Molecular Dynamics Simulation, Nanoemulsion, Drug delivery

Poster 2.8

Evaluation of T-ARMS-PCR for identification of −130G>A in metformin transporter gene SLC47A2/MATE2 using saliva-derived DNA

Hazwanie Hashim1*, Choo Chee Chuan1, Esther Teo Yong Xuan1, Hong Siew Ching1 and Jack Pua Wee Kiet1

1International Medical University, Malaysia

Corresponding author: Dr. Hazwanie Hashim,


Interindividual responses of metformin, a widely used drug for type 2 diabetes mellitus (T2D) have been reported in several studies. SLC47A2 gene that encodes a major transporter protein MATE2 is profoundly associated with the pharmacokinetic and pharmacodynamic variations of drugs including metformin. Of relevance, a single nucleotide polymorphism (SNP) known as −130G>A (rs12943590) is significantly associated with altered glycemic response in T2D patients with metformin. Hence, a reliable and efficient method is essential to identify −130G>A (rs12943590) in clinical laboratory for implementation of personalised medicine in future.


Tetra-primer amplification refractory mutation system polymerase chain reaction (T-ARMS-PCR), a rapid and simple genotyping assay was developed to detect −130G>A (rs12943590). The concentration and purity of saliva-derived DNA were evaluated using a nanophotometer. The absorbance ratio of A260/280 and A260/230 was used to determine DNA quality and purity. Subsequently, the validated method was used to determine the genotype distribution and allele frequencies of −130G>A (rs12943590) in 150 of healthy volunteers.


The assay demonstrated 100% concordance between the direct sequencing and T-ARMS PCR detection. The genotypes distribution of −130G>A namely; AA, AG and GG were 24.7%, 47.3% and 28.0%, respectively in accordance to the Hardy-Weinberg Equilibrium. The lowest detection limit of T-ARMS-PCR assay in this study was 40ng.


In summary, a diagnostic validation of T-ARMS PCR for detection of −130G>A (rs12943590) was successfully performed. T-ARMS-PCR is a rapid and ideal assay for genotype screening in clinical laboratory settings.

Keywords: type 2 diabetes mellitus, T-ARMS-PCR, SLC47A2/MATE2, Single nucleotide polymorphism, Metformin

Poster 2.9

Pro-Inflammatory Cytokines Expression of Carrageenan-Induced Acute Inflammation in Rats Pre-Administered with Freeze-Dried Carica papaya Leaf Juice and Citropten

Norzahirah Ahmad1*, Bee Ping Teh1, Nor Azrina Norahmad1, Siti Khuzaimah Maarof1, Nurul Azreen Mohd Ismail1, Umi Rubiah Sastu@Zakaria1, Nor Azlina Zolkifli1, Azlina Zulkapli2, Tiffiny Ho Chau Dee1, Nabilah Md Razak1, Amirrudin Muhammad1, Bazilah Jusoh1, Wan Abdul Hakim Wan Lokman1 and Mohd Ridzuan Mohd Abdul Razak1
1Herbal Medicine Research Centre, Institute for Medical Research, Ministry of Health (Malasiya), Malaysia
2Medical Research Resource Centre, Institute for Medical Research, Ministry of Health (Malasiya), Malaysia

Corresponding author: Ms. Norzahirah Ahmad,


Immunomodulatory properties of Carica papaya leaf have been previously highlighted in vitro and in vivo. However, repeated dose pre-treatment effect of C. papaya leaf juice and its compounds in inflammatory model is scarce in the literature. Therefore, this study aims to evaluate the anti-inflammatory effects of freeze-dried C. papaya leaf juice (FD-CPLJ) and citropten pre-administrations in a rat model of carrageenan-induced inflammation.


Rats in four treatment groups were pre-administered with distilled water (negative control), FD-CPLJ (2,000 mg/kg body weight (BW)), citropten (20 mg/kg BW) and indomethacin as a positive control (10 mg/kg BW) respectively as a single dose for five consecutive days. On Day 5, inflammation was induced by carrageenan injection to the sub-plantar region of the rat’s left hind paw in the treatment groups. Non-carrageenan stimulated groups were pre-administered with distilled water, FD-CPLJ and citropten. Paw volumes were measured using digital water plethysmometer at certain time intervals (one, three, six and 24 hours) post-induction. The paw tissues were harvested and the expression levels of NOS3, CCL2 and TNF genes were quantitated by quantitative reverse transcription PCR. The expression levels of MCP-1 and TNF-α proteins were determined by Western blot analysis.


Paw volume evaluation shows some reduction in the first and third hour post-induction in groups treated with FD-CPLJ and citropten. The TNF-α expression was slightly downregulated in the citropten pre-treated rats but not in the FD-CPLJ pre-treated rats. The expression of NOS3 gene and the expression of MCP-1 protein was downregulated in FD-CPLJ and citropten pre-treated rats. In contrast to MCP-1 result, the expression of CCL-2 was upregulated in the FD-CPLJ pre-treated rats.


Citropten administration shows anti-inflammatory potential which warrants further investigation at different dosages to further elicit its potential. The contrary results in the gene and protein level for FD-CPLJ-treated group requires further investigation into its mechanism of action.

Keywords: cytokine, Carica papaya, citropten, Inflammation, TNF – α

Poster 2.10

SRS143, A Potential Semi-Synthetic Diterpenoid Lactone for Treatment of Asthma

CheeYI Tan1, Jonathan Chee Woei Lim1*, Hon Liong Soo1, Ibrahim Sulayman1, Screenivasa R. Sagineedu2, Grrace Hui Suan Ng3 and Johnson Stanslas1
1Putra Malaysia University, Malaysia
2International Medical University, Malaysia
3UCSI University, Malaysia

Corresponding author: Dr. Jonathan Chee Woei Lim,


Allergic asthma is one of the most common form of asthma, triggered by inhaling allergens. Histamine, a bioactive compound released by mast cells in response to the allergens, plays an important role in allergic asthma by causing contraction of airway smooth muscles and dilation of capillaries. Our unpublished in-vitro study indicated an Andrographolide analogue (SRS143) potently inhibited histamine release in in vitro mast cell models. Unfortunately, the mechanism of action of SRS143 against the release of histamine and the effect of SRS143 in mouse asthma model have yet to be determined. The aim of our study was to elucidate the antihistamine mechanism and to determine the in vivo anti-inflammatory effect of the analogue in a mouse model of allergic asthma.


In DNP-IgE mediated mast cells (RBL-2H3) models, SRS143 potently inhibit mast cell degranulation and the release of histamine. Subsequently, western blot was performed to investigate the signaling pathway recruiting PI3k, PKC, PLC and AKT. On the other hand, the analogue was also evaluated in a mouse model of House Dust Mite (HDM)-induced allergic asthma. Pulmonary eosinophilia, airway hyper-responsiveness and inflammatory cytokines such as IL-4,-9 and -13 in BAL fluid were measured.


Our protein study shows interesting result whereby AKT and PLC pathway are both targeted by SRS143. We strongly believe that SRS143 targets either or both AKT and PLC pathways to prevent mast cells activation and degranulation. Our in vivo study shows potent antiasthma effect in lung tissue of SRS143 treated allergic mouse asthma model with great decreases in eosinophilia in BAL fluid and improved airway hyper-responsiveness.


SRS143 is a promising Andrographolide analogue to be developed for treatment of allergic asthma.

Keywords: Andrographolide analogues, mast cell activation, Allergic Asthma, degranulation, airway hyper-responsiveness


Poster 3.1

Dexamethasone causes dose-dependent decrease in insulin sensitivity in Wistar albino rats

Rajasekhar Chinta1*, Kokila B Nagaraju2, Raghu Jetti3 and NagendraNayak IM4
1Department of Pharmacology, Melaka Manipal Medical College, Manipal Academy of Higher Education, India
2Department of Pharmacology, KVG Medical College & Hospital,
3Department of Basic Medical Sciences, College of Applied Medical Sciences, King Khalid University, Saudi Arabia
4Department of Pharmacology, Mount Zion Medical College Hospital, India

Corresponding author: Dr. Rajasekhar Chinta,


Long term administration of dexamethasone induces adverse effects such as muscle catabolism, hyperplasia, increased adiposity, and insulin resistance (IR). In view of these data, many authors tried to induce IR with different individual dose ranges with varied induction period. In this study, dexamethasone was used to find the dose to induce maximum insulin resistance in rodents.


A sum of 42 healthy male Wistar albino rats were categorized into six groups of dexamethasone treatment and one control group (n=6/group). In a six-day study period, all treatment groups received respective graded doses of dexamethasone starting from low- (0.5mg/kg and 1mg/kg), intermediate- (2mg/kg and 4mg/kg) and high-dose (8mg/kg and 16mg/kg).


Graded doses of dexamethasone treatment for six days produced hyperglycemia and hyperinsulinemia in a dose-dependent manner and the maximum effect was noted with high doses of dexamethasone compared to intermediate and low (p<0.05). The profound reduction in insulin sensitivity was caused by high-doses of dexamethasone treatment evidenced by sustained elevation of homeostatic model assessment of insulin resistance (HOMA-IR) which was strongly associated with declined homeostatic model assessment of insulin sensitivity (HOMA-IS), The peripheral sensitivity indices, Gutt and Matsuda, were markedly elevated in high dexamethasone groups compared to intermediate- and low-dose groups (p<0.05). Serum lipids and creatinine (p<0.05), whereas high density lipoprotein cholesterol (HDL-CH) was markedly reduced and resulted in the subsequent rise in atherogenic index (AGI) (p<0.05). Moderate to severe glycosuria and ketonuria were noted in intermediate- and high-dose treatment groups only. However, there is no significant difference in metabolic effects between 8mg/kg and 16mg/kg treatments (p>0.05).


Administration of 8mg/kg dexamethasone for six days would be sufficient for the induction of maximum insulin resistance in Wistar albino rats.

Keywords: HOMA, Insulin Resistance, Hyperglycemia, hyperinsulinemia, Metabolic effects

Poster 3.2

Modulation of glucose uptake and insulin sensitivity in insulin resistant in vitro Alzheimer’s Disease model by vitamin D and E

Nafissa Nadia Ibrahim1*, Huzwah Khaza’ai1, Amirah Salwani Zaulkaffli1 and Norshariza Nordin1
1Department of Biomedical Sciences, Faculty of Medicine and Health sciences, University of Putra Malaysia, Malaysia

Corresponding author: Miss. Nafissa Nadia Ibrahim,


Currently, there is a widening recognition that impaired insulin signaling which causes a brain-specific form of diabetes contributes to the pathogenesis of Alzheimer’s Disease (AD). Previous study has reported that high concentration of insulin can alter expression of insulin signal transduction cascade such as insulin receptor (IR), glucose transporter (GLUT4), phosphatidylinositol 3-kinase (PI3K) and glycogen synthase kinase 3 (GSK3) which lead to inhibition of glucose uptake into the cell. Progressive derangements of brain glucose utilization due to insulin insensitivity cause the impairment of glucose homeostasis, hence increasing the oxidative stress which leads to the pathogenesis of AD. Vitamin D has been recognized to have a direct beneficial effect on insulin action by stimulating the expression of insulin receptors. Meanwhile, vitamin E possesses a neuroprotective effect by scavenging oxidative stress effectively. This study aims to elucidate the effect of vitamin D and E in modulating insulin signaling and AD markers in insulin resistant in vitro AD model.


SK-N-SH (neuroblastoma) was induced with high insulin prior to assess the mRNA expression of insulin signaling markers (IR, GLUT4 and PI3K), AD markers (TAU and GSK3β) and glucose uptake level. Treatment of vitamin D, E and a combination treatment were introduced for 24 hours upon completion of insulin induction.


Improvement of insulin signaling pathway was observed upon treatment of vitamin D alone by significantly increase the level of IR, GLUT4 and PI3K, and glucose uptake level. Reduced expression of both AD markers was observed upon treatment with vitamin D. However, vitamin E and a combination treatment of both vitamins showed insignificant changes in insulin signaling and AD markers.


Vitamin D and E may demonstrate as an agent that slows the progression of Alzheimer Disease caused by insulin resistance in the brain.

Keywords: Insulin Resistance, Alzheimer’s disease, Vitamin E, Vitamin D, neurodegenerative

Poster 3.3

Evaluation of Total Phenolic, Flavonoids, and Antioxidant Activities of Crude and Fractionated Extracts of Selected Local Medicinal Plants

Phaik Har Yong1*
1Mahsa University, Malaysia

Corresponding author: Dr. Phaik Har Yong,


Recent studies have suggested that the development of metabolic syndrome complications could be reduced by the intake of natural antioxidants from plants. Medicinal plants have been used as traditional medicine for the treatment of various diseases related to oxidative stress by local populations in Malaysia. Hence, this study hypothesizes that the selected medicinal plant extracts would be able to demonstrate significant levels of phenolic and flavonoid compounds which possess anti-oxidant activities. The objective of this study was to evaluate the antioxidant potential of Centella asiatica, Phyllanthus urinaria, Folium mori, Rhinacanthus nasutus, Alternanthera sessilis and Strobilanthes crispus.


Selected local plants were processed and macerated with absolute methanol. The solvent extracts were rotary evaporated, subjected to fractionation with different solvents in the following order: water, hexane, chloroform, ethyl acetate and n-butanol. The antioxidant activities of extract and fractions were evaluated through Ferric Reducing Antioxidant Power (FRAP), 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) free radicals scavenging assays. Folin-Ciocalteu and aluminium chloride methods were used to quantify the phenolic, flavonoid contents of crude extracts and fractions.


P. urinaria demonstrated considerable levels of phenolic, flavonoids and antioxidant activities across all extract fractions. In contrast, only a few selected fractions of other plant samples showed high levels of activities. These were particularly of ethyl acetate, n-butanol and chloroform fractions which possessed high levels of phenolic, flavonoids and antioxidant activities. Ethyl acetate fraction of C. asiatica displayed the highestphenolic, flavonoid contents and antioxidant activities when compared to the other plant samples. In general, the phenolic (r = 0.875, p<0.001) and flavonoid contents (r = 0.527, p<0.001) were positively correlated with total antioxidant activities.


The study showed that the selected medicinal plants, particularly P. urinaria possess high antioxidant potential and could be a potential natural source of antioxidant.

Keywords: Antioxidants, medicinal plants, Phenolic, Flavonoids, Fractionation

Poster 3.4

Hyperglycaemia in pregnancy disturbs maternal-foetal well-being and alters placental function: role of inflammatory markers fkn and rank/rankl/opg system

Noor E. Hashim1*, Mardhiah Tahir1, Intan S. Zulkafli1, Rasheeda Mohd. Zamin1 and Normadiah M Kassim1
1University of Malaya, Malaysia

Corresponding author:  Dr. Noor E. Hashim,


Inflammation have been implicated for various diabetic complications. Fractalkine (FKN) and receptor activator of nuclear factor-kappaB ligand (RANKL)/RANK/osteoprotegerin (RANKL/RANK/OPG) systems are inflammatory markers suspected to be responsible for the changes seen in diabetic pregnancy. This study aimed to examine the effects of hyperglycaemia on pregnancy and investigate the role of these markers in the disease process.


Pregnant female Sprague-Dawley rats were injected with 45mg/kg BW of Streptozocin on gestational day (GD) 7 to induce diabetes followed by 10% glucose drink for 24h to prevent hypoglycaemic fatality. Control animals were injected with 1ml of citrate buffer vehicle. Diabetes was confirmed on GD15. Maternal weight and food intake were regularly monitored. Blood (maternal & foetal) and placenta separated into labyrinth and junctional zones were collected for morphological, biochemical and molecular analyses at GD15 and GD21.


Maternal hyperglycaemia induced hyperphagia (p<0.05) and impeded maternal weight gain (p<0.05). Both groups were able to carry pregnancy to term but the hyperglycaemic group (HG) demonstrated trend towards increased foetal resorption at GD21 (13.6%, p=0.08) and 13.2% reduction of foetal weight (p<0.05). Although placenta weights were unchanged in all groups, feto-placental ratio dropped 15.8% in the HG group at GD21 (p<0.05). Morphologically, all placenta showed no significant differences in weight or thickness at both time points. Placenta zonal variations were observed in the mRNA expression of RANK, OPG, FKN and Vascular Endothelial Growth Factor A (VEGFa) but no hyperglycaemic effect was found.


This study demonstrated that maternal hyperglycaemia is detrimental in pregnancy through disruption of foetal survival (increased foetal resorption), disturbed maternal well-being (via altered eating pattern) and placental insufficiency through reduction of feto-placental ratio. Although expression of the inflammatory markers showed no significant role in the altered physiological milieu, more detailed studies are needed to delineate the exact role of these markers in hyperglycaemic pregnancy.

Keywords: Maternal hyperglycaemia, Placenta, Pregnancy, Inflammatory markers, rat model

Poster 3.5

Design, Synthesis, In vitro Biological Evaluation, and Molecular Docking Studies of a Series of p-Toluene Sulfonylurea-Based Hybrids as a New Class of α-Glucosidase Inhibitors

Vasudeva Rao Avupati1*, Yap W. Sheng1, Diong Q. Cyrus1, Wong Z. Feng1 and Mallikarjuna Rao Pichika1
1International Medical University, Malaysia

Corresponding author: Dr. Vasudeva Rao Avupati,


α-Glucosidase is an intestinal membrane-bound enzyme that catalyze the final step in the digestion of carbohydrates. It cleaves the glycosidic bond of the oligosaccharides to release glucose and its inhibition results in a delayed glucose absorption and thereby inhibiting postprandial hyperglycemia and hyperinsulinemia in type 2 diabetic patients. α-Glucosidase inhibitors have an additive effect on sulfonylureas when used in combination, and also diminish the insulinotropic and weight-increasing effects of sulfonylureas.


A series of novel p-toluene sulfonylurea-based hybrids have been synthesized and characterized by FTIR, 1H NMR, 13C NMR and LC mass spectral analysis. All the compounds were evaluated for their in vitro α-glucosidase inhibitory potential. Molecular docking studies have also been performed using the Schrödinger software “Glide module”.


Among the tested compounds for α-glucosidase inhibitory potential, compounds (E)-N-((4-(3-(3-bromophenyl)acryloyl)phenyl)carbamoyl)-4-methylbenzenesulfonamide and (E)-N-((4-(3-(3-fluorophenyl)acryloyl)phenyl)carbamoyl)-4-methylbenzenesulfonamid exhibited significant inhibition at IC50 value 1.89 µM and 2.10 µM respectively.


We exposed p-toluene sulfonylurea-based hybrids as a new class of α-glucosidase inhibitors. The results revealed the positive contribution of halogen substituents on the phenyl ring towards the observed activity. The observed activities may also be due to the sulfonamido group and α,β-unsaturated ketone moieties forming part of the basic structure of these molecules.

Keywords: Hybrid molecules, p-Toluene sulfonylureas, α-Glucosidase inhibitors, Molecular docking analysis, Schrödinger software suite

Poster 3.6

A Pilot Study on the Association between SLCO1B1 rs4363657 Polymorphism and Muscle Adverse Events in Adults with Newly Diagnosed Dyslipidaemia who were Prescribed a Statin: The Malaysian Primary Health Care Cohort

Meor Fairuz Rizal Meor Anuar Shuhaili1, Boon How Chew2, Intan Nureslyna Samsudin1, Hejar Abdul Rahman3, Johnson Stanslas4, Shariful Hasan4, Zalinah Ahmad1 and Subashini C. Thambiah1*
1Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia
2Department of Family Medicine, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia
3Department of Community Health, Faculty of Medicine and Health Sciences, Putra Malaysia University, Malaysia
4Department of Medicine, Faculty of Medicine and Health Sciences, University Putra Malaysia, Malaysia

Corresponding author: Dr. Subashini C. Thambiah,


Statin, the first-line treatment for dyslipidaemia, may have suboptimal adherence due to its associated muscle adverse events. This data however, remains limited, due to under-reporting, mis/underdiagnoses or statin-associated muscle adverse events are generally rare. The aim of this study is to determine the association of serum creatine kinase (CK) and rs4363657 polymorphism of SLCO1B1 gene with statin-associated muscle adverse events among dyslipidaemia subjects.


This was a prospective cohort study at government health clinics involving newly diagnosed adults with dyslipidaemia. Muscle adverse events were recorded based on the patient’s complaint after a month on statin (lovastatin or simvastatin). Biochemical analyses including CK, fasting lipid profile, apo A1, apo B were taken at baseline and follow-up. Genetic profiling was performed for rs4363657 polymorphism of SLCO1B1 gene.


A total of 118 subjects participated. Majority were Malay (72%) males (61%) with a mean age of 49 ± 12.2 years old and prescribed lovastatin (61.9%). There was a significant association between types of statin and muscle adverse events (p = 0.0327); frequency of muscle adverse events was higher with lovastatin (15.2%). No significant association noted between CK and muscle adverse events (p = 0.5637). The rs4363657 polymorphism of SLCO1B1 gene was significantly associated with muscle adverse events (p < 0.0001). The frequency of muscle adverse events was highest in CC genotype (100%); TC and TT (wild type) genotypes were 42.31% and 3.57%, respectively. Subjects with TC and CC genotypes were 26.24 and 357.96 times more likely to exhibit muscle adverse events compared to subjects with normal SLCO1B1 gene, respectively (p < 0.0001).


Statin-associated muscle adverse events were significantly higher in subjects on lovastatin compared to simvastatin. rs4363657 polymorphism of SLCO1B1 gene was a significant risk factor for statin-associated muscle adverse events in this study.

Keywords: Dyslipidemia, statin, muscle adverse events, SLCO1B1, rs4363657, Creatine Kinase, Simvastatin, Lovastatin


Poster 4.1

Therapeutic effect of Phaleriamacrocarpa (leaf) methanolic extract in management of neurodegenerative diseases on SH-SY5Y cells and Javanese medaka (Oryziasjavanicus) model

Adamu Abdul Abubakar1, and Hassan Maina Ibrahim2*
1Department of Veterinary Surgery and Radiology, Usmanu Danfodiyo University Sokoto, Nigeria
2Department of Veterinary Physiology and Biochemistry, Usmanu Danfodiyo University Sokoto, Nigeria

Corresponding author:  Adamu Abdul Abubakar,


Neurodegenerative diseases refer to physiological or biochemical changes leading to the malfunction or death of nerve cells present in the brain and spinal cord that are characterized by loss of function or alteration of sensory functions. Several diseases that affect neurons such as Parkinson’s disease (PD), Alzheimer’s disease (AD), Multiple Sclerosis (MS) and amyotrophic lateral sclerosis (ALS) may be grouped under this abnormal condition in which the etiological causes may differ but commonly associated with mitochondrial dysfunctions, excite-toxicity and apoptosis. Current treatments for those neurodegenerative diseases are not effective and caused many side effects. Thus, the search for alternative medicines is in high demand.


The aim of this study was to evaluate the reactive oxygen inhibitory and neuroprotective properties of Phaleriamacrocarpa (leaf) extract on SH-SY5Y cells and adult Javanese medaka (Oryziasjavanicus).


Antioxidant using dichlorofuoresence diacetate (DCF-DA) assay on SH-SY5Y cells revealed high activities of Phaleriamacrocarpa (leaf) at IC50 of 146.66±0.1 µg/mL. Sub-acute and chronic toxicity test of the plant extract on adult Javanese medaka (Oryziasjavanicus) demonstrate low effect with LC50 of 154.38 ± 0.6 and 41.29 ± 0.9 mg/mL. Neuroprotective test using acetyl-cholinesterase, butyryl- cholinesterase and propionyl-cholinesterase assays revealed significant differences (P<0.05) between the treated and non-treated groups. Present of bioactive compounds such as the flavonoid may be among the major secondary metabolite that contributed in increase activities of this plant extract.


High antioxidant activities and low toxicity effect of this plant indicate its high potential benefit to be used as and natural medicine/supplements.

Keywords: Antioxidant, Phaleriamacrocarpa, Javanese medaka, Neuroprotective

Poster 4.2

Thymoquinone-loaded nanostructured lipid carrier improved spatial learning, memory and exploratory behaviour in Alzheimer’s disease animal model

Fatin Hannani Zakarial Ansar1, Latifah Saiful Yazan2*, Ng Wei Keat1, How Chee Wun3, Rasedee Abdullah3, Mohamad Aris Mohd Moklas4 and Cheah Pike See4
1Laboratory of Molecular Biomedicine, Institute of Bioscience, University Putra Malaysia, Malaysia
2Department of Biomedical Sciences, Faculty of Medicine and Health sciences, University of Putra Malaysia, Malaysia
3Universiti Putra Malaysia, Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience, University Putra Malaysia, Malaysia
4Department of Human Anatomy, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia

Corresponding author:  Dr. Latifah Saiful Yazan,


Alzheimer’s disease (AD) is a devastating neurodegenerative disease with progressive and irreversible damage to thinking, memory and language. Its viable cure remains unavailable. Research on bioactive compounds from natural products with neuroprotective effects and low toxicity is on demand. Thymoquinone (TQ), a compound from Nigella sativa possesses remarkable antioxidant properties is able to protect cells from degradation. However, TQ has low solubility in blood and poor oral bioavailability. Hence, nanostructured lipid carrier (NLC) has been developed as a drug delivery vehicle to overcome the limitations (herein referred as TQ-NLC). This study aimed to evaluate the neuroprotective effects of TQ-NLC as a potential candidate drug for the management of AD in vivo.


Sprague Dawley rats were fed with high fat cholesterol diet as a model of Alzheimer’s disease, treated with 12.5, 25 and 50mg/kg of TQ-NLC and TQ, were subjected to spatial learning and memory using Morris water maze. Lipid profile level was also determined using an automated hematology analyzer.


The animals treated with 25mg/kg of TQ-NLC showed an increase in the time spent at the targeted quadrant and reduced in the total cholesterol compared to the negative control (untreated) (p<0.05). In addition, the animals treated with 25mg/kg of TQ-NLC showed shorter escape latency in comparison to the negative control (untreated) but it was not statistically significant.


Treatment with TQ-NLC improved spatial learning and memory of the animals and their cholesterol level.

Keywords: Alzheimer’s disease, Thymoquinone, NLC (nanostructured lipid carriers), TQ-NLC, high fat cholesterol diet

Poster 4.3

Neuro-recovery of vitamin E in high glutamate-induced oxidative stress neural cells derived from embryonic stem cell culture model

Nurliyana Najwa Md Razip1, Afifah Abd Jalil1, Norshariza Nordin1, 2, Amirah Salwani Zaulkffli3, Nur’Izzati Mansor1, 2 and Huzwah Khaza’ai1*
1Department of Biomedical Sciences, Faculty of Medicine and Health sciences, University of Putra Malaysia, Malaysia
2Genetics and Regenerative Medicine Research Centre, Faculty of Medicine and Health Sciences, Putra Malaysia University, Malaysia
3Department of Nutrition and Dietetics, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia

Corresponding author: Dr. Huzwah Khaza’ai,


Excitatory glutamatergic of neurotransmitter is essential for neuronal plasticity in the brain. However, excessive glutamate-binding activity over-stimulates the postsynaptic receptors in neuron cause excitotoxic and oxidative stress, which is associated with the pathogenesis of Alzheimer’s Disease (AD). High level of reactive oxygen species explicates neuro-injury due to lack of antioxidant. Further study elucidates the potential of vitamin E against the neuronal injury mechanisms.


Neural derived embryonic stem cells were exposed to a toxic concentration of glutamate (60 mM) enough to demonstrate the injured neuronal model. The potential of vitamin E [tocotrienol rich fraction (TRF) and alpha-tocopherol (alpha-TCP)] in modulating oxidative stress and recovery process upon glutamate toxicity was elucidated. The expression of glutamate receptors (N-methyl-d-aspartate receptors, NMDAR and kainate receptors) and neuron injury marker (neuron-specific enolase, NSE) were quantified with real-time polymerase chain reaction (RT-PCR) as a biomarker of the recovery process.


Supplementation of vitamin E at 100, 200 and 300 ng/mL reduced 90% rate of overstimulation NMDA-1 (GluN-1) compared to positive control with fold ratio 0.347±0.03, 0.195±0.04 and 0.083±0.01, respectively. Gluk1 is a kainate receptor reduced with the fold ratio of 0.594±0.16, 0.320±0.02 and 0.291±0.07 respectively compared to positive control. NSE decreased significantly with the fold ratio at 0.525±0.03, 0.514±0.02 and 0.605±0.11 respectively. Meanwhile, alpha-TCP treatment at concentration of 100, 200 and 300 ng/ml on glutamate toxicity model indicated the significant reduction of GluN-1 (0.646+0.06, 0.583+0.01 and 0.530+0.09) respectively, Gluk1 indicates significant reduction at 100 and 200 ng/ml at fold ratio of 0.614±0.09 and 0.502±0.04 and NSE (0.468±0.08, 0.460±0.1 and 0.782±0.12) fold ratio.


Both isomers of vitamin E in the form of tocotrienols and tocopherols are suggested as a potent antioxidant that quenching radicals hence counter-regulate the excessive glutamate stimulated receptors which help to reduce AD risk.

Keywords: Glutamate, neuronal injury, Oxidative Stress, Alzheimer ‘ s disease, Vitamin E

Poster 4.4

Neuroprotective effect of Zingiber zerumbet ethyl acetate extract in paraquat-induced Parkinsonism rats via its anti-inflammatory activity

Farah Wahida Ibrahim1*, Nur Nabila Zakaria1, Umi Noraashikin Zainudin1, Mazlyzam Abdul Latif1 and Asmah Hamid1
1National University of Malaysia, Malaysia

Corresponding author: Dr. Farah Wahida Ibrahim,


The progressive loss of dopaminergic neurons in Parkinson’s disease is associated with exposure to environmental toxins such as herbicide. In this study, the neuroprotective effect of Zingiber zerumbet extract was tested in paraquat-induced Parkinsonism rats.


Fifty male Sprague-Dawley rats (180-240g) were divided into five groups: normal control, positive control (N-acetyl cysteine + Paraquat, PQ), negative control (PQ only), 200mg/kg Zingiber zerumbet + PQ, and 400mg/kg Zingiber zerumbet + PQ. Ethyl acetate extract of Zingiber zerumbet were given orally for 19 consecutive days, and PQ was administered intraperitoneally (10mg/kg/day) on day 8th-12th of the treatment regime. The level of nuclear factor Kappa β (NFκβ), interleukin 1β (IL-1β), inducible nitric oxide synthase (iNOS), and α-synuclein proteins of the striatum were determined via enzyme-linked immunosorbent assay. The total protein content in serum and striatum homogenate was measured via the Bradford method.


In general, the levels of total protein contents in serum and striatum homogenate, as well as the level of iNOS, were significantly lower in the Zingiber zerumbet-treated groups (200 and 400mg/kg) compared to the negative control group (p<0.05). The results also revealed that the level of NFκβ was significantly lower in rats receiving 400mg/kg of Zingiber zerumbet extract compared to the negative control rats (p<0.05). There was also a significantly higher level of α-synuclein in both normal control and 400mg/kg Zingiber zerumbet-treated groups compared to the negative control group (p<0.05). The substantia nigra of Zingiber zerumbet-treated rats at both doses showed significantly higher dopaminergic neuronal number compared to the negative control group (p<0.05). Tremor scores of rats in Zingiber zerumbet-treated groups (200 and 400mg/kg) showed progressive reductions at PQ post-induction phase (day 13th – 19th).


Ethyl acetate extract of Zingiber zerumbet able to provide neuroprotection through its anti-inflammatory activity in PQ-induced Parkinsonism rats.

Keywords: Zingiber zerumbet, Paraquat, Parkinson’s disease, Neuroprotection, anti-inflammatory activity

Poster 4.5

Influence of Gestational Diabetes on Cognitive Function of the Adolescent Male/Female Offsprings

Cini M. John1, 2, 3, Atish Prakash4, Nur Intan S. Yusof2, Aishah Adam2, Rajesh Ramasamy3 and Fazlin M. Fauzi2*
1University of Calgary, Canada
2Universiti Teknologi MARA Puncak Alam, Malaysia
3Putra Malaysia University, Malaysia
4Johns Hopkins Medicine, United States

Corresponding author: Dr. Fazlin M. Fauzi,


Maternal gestational diabetes has been proposed to negatively affect the cognitive abilities of the offspring. The underlying pathogenic mechanisms behind the abnormal cognitive risk profile in offsprings are unknown, but epigenetic changes induced by exposure to maternal hyperglycaemia and insulin resistance during gestation are implicated.


In this current study, 10-11 weeks old male/female offsprings of GDM rats and normal Sprague Dawley rats were used. Learning and memory functions, serum BDNF levels, structural changes in hippocampus were examined. To understand the immunological changes contributing to the functional effects in the brain, we analyzed subsets of T and B lymphocytes from splenocytes, circulating cytokines and anti/pro-inflammatory markers from brain.


Our results show that mild impairment in learning and memory functions were observed in GDM offsprings, where they clocked a longer escape latency time and shorter time spent in target quadrant than the control groups however no significant difference were observed between male and female groups. Low BDNF levels were observed only in male offsprings. No structural changes were found in the normal layer organization in the hippocampus. Immune cell characterization revealed no significant changes in T or B cell populations, besides FOXP3+ regulatory T cells, which was high in the female offsprings along with anti-inflammatory cytokines IL-5, IL-10 and TGF-b. Pro/anti inflammatory markers IL-1beta, IL-12, IFN-γ and TNF-α, primary transcription factors NF-Kb didn’t show any changes between groups. There was a high expression of phosphoprotein p53 and IL-6 in male and female groups in the brain tissue of GDM offsprings.


In conclusion, these data demonstrate that mild cognitive impairment observed in the offsprings of GDM rats and were similar in male and female, however female offsprings are favoured while maintaining an anti-inflammatory environment in the brain tissues and circulation with expanded Treg cell population. It is important to further define and investigate sex differences in GDM influenced offspring’s metabolic and immune functions, which help to improve and optimize cognitive health and aid future research.

Keywords: cognitive functions, GDM, male/female offspring, BDNF, Immune changes

Poster 4.6

The Effect of Mindfulness Meditation on Lowering Stress Level in Medical Students

Dian Wulandari1*, Darmawati A. Indraswari2, Yosef Purwoko2 and Ainun R. Gumay2
1Faculty of Medicine, Diponegoro University, Indonesia
2Department of Physiology, Faculty of Medicine, Diponegoro University, Indonesia

Corresponding author: Miss. Dian Wulandari,


Many studies have reported that the prevalence of psychological stress among medical students during education was high. However, there has not been an effective and efficient method to help medical students coping with stress in that particular situation. Mindfulness meditation has been practiced worldwide in any level of society on lowering stress. This study aims to determine the effect of mindfulness meditation on lowering stress level in medical students.


This is an experimental study with pre- and post-test control group design. Thirty medical students of first year were included in the study were divided into two groups and filled the cognitive test anxiety scale (CTAS) and Hamilton anxiety rating scale (HARS) as the pretest. The intervention group of fifteen students did a mindfulness meditation program in fourteen days using meditation application in android/ios. While the control group of fifteen students did not do a mindfulness meditation program in fourteen days or any similar activities. Post-test for both groups were applied after the intervention with CTAS and HARS. The stress level were calculated using the T test and Wilcoxon test.


There was a significant decrease in both CTAS and HARS after doing mindfulness meditation in intervention group compared to before doing mindfulness meditation. Meanwhile there was not a significant decrease in both CTAS and HARS in control group.


Mindfulness meditation helps lowering the stress level in medical students.
Keywords: stress, mindfulness meditation, medical students, CTAS, HARS.

Keywords: stress, mindfulness meditation, Medical student, CTAs, HAR


Poster 5.1

Solution Enhanced Dispersion by Supercritical Carbon Dioxide (SEDS) Co-precipitation as a Propitious Strategy for “Bottom-up” Andrographis paniculata Powder Production with Improved Dissolution

Sin Yee Lee1, Luqman Chuah Abdullah1, Russly Abdul Rahman1, Faridah Abas1 and Gun Hean Chong1*
1Putra Malaysia University, Malaysia

Corresponding author:  Dr. Gun Hean Chong,


Plants have been the primary source of natural product drug discovery besides being home remedies for various ailments. Nevertheless, poor aqueous solubility and dissolution rate of andrographolide from Andrographis paniculata have become the liabilities to their therapeutic effects.


Two main strategies, (a) reducing crystallinity and (b) increasing surface area available for dissolution had been applied through Solution Enhanced Dispersion by Supercritical Carbon Dioxide (SEDS) co-precipitation for improving the dissolution of andrographolide from A. paniculata extract in Simulated Intestinal Fluid (SIF), pH 7.4. Sticky crude A. paniculata extract containing 16 % of andrographolide had been co-precipitated with Eudragit L100-55 with the SEDS working parameters set constant as follows: 150 bar, 40 ̊C, 15 L/min of CO2 flow rate (1 bar, 25 ̊C), 0.5 mL/min of liquid feed flow rate, and 25 mg/mL of crude A. paniculata extract.


Through SEDS co-precipitation with mass ratio of Eudragit L100-55 to A. paniculata at 2:25, dissolution rate of andrographolide in target environment, SIF, pH 7.4 had been greatly improved (0.06 mg/mL release in 45 min) compared to SEDS precipitated A. paniculata extract powder (0.06 mg/mL release in 90 min). Increment of mass ratio of Eudragit L100-55 to A. paniculata from 2:25 to 6:25 had resulted in higher release and dissolution rate of andrographolide in SIF, pH 7.4 (0.09 mg/mL release in 30 min). SEDS co-precipitates with mass ratio of Eudragit L100-55 to A. paniculata at 6:25 was also found to have the highest cytotoxicity (LC50 = 46.46 µg/mL) as compared to crude A. paniculata extract (LC50 = 150.45 µg/mL) and SEDS precipitated A. paniculata extract powder (LC50 = 160.06 µg/mL).


SEDS co-precipitation had improved dissolution of andrographolide from A. paniculata that resulted in increment of its bioactivity or cytotoxicity.

Keywords: SEDS, co-precipitation, andrographolide, Eudragit L100-55, dissolution

Poster 5.2

Computational Studies of Steroidal 5α-Reductase Inhibitors using QSAR, Molecular Docking and Pre-ADME Prediction

Richa Dhingra1, Neelima D Passi1, Manav Malhotra2, Monika1, Tilak Raj Bhardwaj3

1University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh-160014, India
2M.K Drugs, F-10 Focal point, Industrial Area, Derrabassi-140587, India.

3School of Pharmacy and Emerging Sciences, Baddi University, Baddi, H.P.-173205, India


5α-Reductase (5AR) an NADPH dependent enzyme is expressed in most of the epithelial cells in prostate gland. Significant efforts have been made in order to develop 5AR inhibitors (5ARI) to treat Benign Prostatic Hyperplasia (BPH). We have made an attempt to design molecules against 5AR enzyme using Quantitative structure activity relationship (QSAR), docking and Pre-ADME (Absorption, distribution, metabolism and excretion) etc.


In the present study, two and three dimensional (2D and 3D) QSAR models have been developed for steroidal 5α-Reductase Inhibitors (5ARI) using Vlife MDS 4.6 with extensive internal and external validation process. Data set of 3β-substituted amides of 17a-Aza-D-homo-4-androsten-17-one derivatives were used with their reported IC50 value. pIC50 was considered as dependent variable and the remaining descriptors as independent variables to elucidate the structural properties required for 5ARI.  Compounds were synthesized utilizing relevant information from QSAR and were characterised spectrophotometrically. Molecular docking studies of synthesized steroids was carried out to identify the binding orientation and the receptor ligand interactions responsible for exhibited activity with protein having PDB ID: 4AT0 using Biopredicta tool in Vlife software. To gain more insights into the molecular mechanism of action of these steroidal derivatives, in silico ADME studies were also performed.


Four featured 2D QSAR pharmacophore PLS based model for 5AR correlated the topological interactions (SsOHE-index); semi empirical (Quadrupole2) and physicochemical descriptors (Mol. Wt, Bromines Count, Chlorines Count) with 5AR inhibitory activity with  high correlation coefficient (r2=0.98, q2=0.84; F=57.87, pred r2=0.88). 3D-QSAR studies revealed the importance of less bulkier (steric interactions) and more electronegative group at different positions on steroidal nucleus for cytotoxicity activity. Ligand receptor interaction indicated the hydrophobic and hydrogen bonding of the ligands (newly synthesied molecules) with amino acid residues ALA 63A, THR 60A, and ARG 456A of 4AT0 protein at the hinge region. All the novel compounds showed good absorption, plasma protein binding and followed Lipinski rule of five.


The well validated 2D PLS, 3D kNN models provided us with the important structural physicochemical descriptors for identification of more potent 5ARI inhibitors with different chemical scaffold. Molecular docking and in silico ADME studies showed mechanistic scope for further lead optimization that helps in designing of novel 5ARIs.


5α-reductase inhibitor, Benign Prostatic Hyperplasia, Ligands, Molecular Docking, QSAR.

Poster 5.3


Farah Dayana Rosli1*, Noor Hashida Hashim2, Yusmin Mohd Yusuf3, Khairul Osman4, Siti Fatimah Ibrahim5 and Nurul Kabir6
1Institute of Graduate Studies, University of Malaya, Malaysia
2Biology Division, Center for Foundation Studies in Science, University of Malaya, Malaysia
3Centre for Research in Biotechnology for Agriculture (CEBAR), University of Malaya, Malaysia
4Faculty of Allied Health Sciences, National University of Malaysia, Malaysia
5Faculty of Medicine, National University of Malaysia, Malaysia
6Institute of Biological Sciences, University of Malaya, Malaysia

Corresponding author: Miss. Farah Dayana


Thymoquinone (TQ), the main constituent of the volatile oil derived from Nigella sativa has demonstrated therapeutic potentials against various diseases while nicotine is an active component in cigarette known to be detrimental towards sperm quality. This study was conducted to assess the effects of TQ in ameliorating the injurious state in sperm of nicotine treated rats.


Adult male Sprague-Dawley rats were divided into four equal groups: control group received normal saline orally for 60 days; nicotine group was subcutaneously injected with 5 mg/kg/day nicotine for 30 days and then given normal saline for next 30 days; TQ group was given normal saline for 30 days followed by TQ 5 mg/kg/day for 30 days; nicotine-TQ group, received 5 mg/kg nicotine for 30 days, followed by TQ 5mg/kg for 30 days. The sperm count, motility and mitochondrial function were evaluated.


Sperm count was significantly lower (p<0.05) in the nicotine group (26.72 ± 1.64) in comparison to the control (31.75 ± 0.80). Whereas the number of sperm was significantly higher in the nicotine-TQ group (30.97 ± 0.88) compared to the nicotine group. Motility evaluation demonstrated similar pattern with number of motile sperm was higher in the nicotine-TQ group (85.02 ± 2.24 %) compared to the nicotine group (66.24 ± 1.01 %). In mitochondrial function assessment, nicotine-TQ group (79.08 ± 0.74 %) showed higher (p<0.05) MitoTracker positive sperm percentage compared to nicotine group.


Thus, the present study suggests the potential of TQ in ameliorating nicotine-induced sperm damage.

Keywords: Thymoquinone, Nicotine, Sperm Count, motility, Mitochondria

Poster 5.4


Amirah Baharin1*
1University of Malaya, Malaysia

Corresponding author: Mrs. Amirah Baharin,

Morphine is one of the opioid drugs that have been long used in the field of medicine to treat severe pain. Excessive or repeated use of the drug can contribute to infertility. On the other hand, dates or P. dactylifera contains carotenoid, cholesterol, vitamins and energy that possesses an antioxidant and anti-infertility effects. The aim of this study was to evaluate the effects of P. dactylifera on the histomorphometric of male reproductive organs (testis, seminal vesicle (SV) and prostate gland (PG) of morphine-treated rat. Adult male Sprague Dawley rats age 7-9 weeks old, 200-250g body weight (BW) were randomly divided into 4 groups: i. Group 1(control group) was force-fed with distilled water, 1ml/kg BW for 35 days, ii. Group 2 was intramuscularly (i.m) injected with Morphine, 20mg/kg BW for 7 days followed by distilled water for 28 days, iii. Group 3 was force-fed with distilled water for 7 days followed by crude P. dactylifera extract, 200mg/kg for 28 days and iv. Group 4 was injected (i.m.) with morphine, 20mg/kg BW for 7 days followed by force-fed of crude P. dactylifera extract, 200mg/kg for 28 days. On day 36, the rats were sacrificed. The testis, SV and PG were extracted for histomorphometric study. The results indicated that morphine exposure induced apoptosis in the testicular germ cells and reduction in the height of columnar epithelial cells of the SV and PG as compared to other treatment groups. The effects were however, noted to be counteracted by the P. dactylifera whereby an improvement in the histomorphometric of the testis, SV and PG was noted similar to that of found in the control group. This ameliorating effect indicated that dates contains a potential therapeutic agent which could be used in treating male infertility.

Keywords: Phoenix dactylifera, Morphine, testis, seminal vesicle, Prostate gland

Poster 5.5

Trigonella-Foenum Graecum L. Seed Mucilage-Based Diclofenac Sodium Mucoadhesive Microspheres

Senthil Adimoolam1* and Venkata S. Appala Raju Velaga1
1Mahsa University, Malaysia

Corresponding author:  Dr. Senthil Adimoolam,


The study was aimed to discover the properties of Trigonella foenum graecum L. seed mucilage (TFGSM) in the formulation of mucoadhesive microspheres of diclofenac sodium to prolong the residence time at the site of absorption.


TFGSM-based mucoadhesive microspheres of diclofenac sodium were prepared by ionotropic gelation technique by using different polymer ratios of TFGSM and sodium alginate. Eight formulations (F1 to F8) of TFGSM-based mucoadhesive microspheres of diclofenac sodium was formulated and characterized by its percentage yield, drug entrapment efficiency, mucoadhesivity, swelling properties and percentage drug release.


The effects of polymer ratios (TFGSM to sodium alginate) on drug entrapment efficiency, mucoadhesivity, swelling index and percentage drug release over 8 h were investigated. The drug entrapment efficiency of all eight formulations was within the range of 78 to 98% with sustained in vitro release of over 8 h. The in vitro drug release of these microspheres followed controlled release (zero-order & Fickian diffusion). The microspheres possessed good swelling properties and mucoadhesive properties. The FTIR spectrum suggested there were no interactions between drug and excipient used in formulation.


The study shows that the TFGSM-based microspheres formulated were able to prolong gastric residence time by swelling slowly in stomach and adhering to the stomach mucosa which then will subsequently move to the intestine and swells more allowing the release of drug. Thus, TFGSM serves as a potential mucoadhesive excipient in the formulation of controlled-release mucoadhesive microsphere.

Keywords: microspheres, Trigonella foenum graecum L, Seed mucilage, mucoadhesion, diclofenac sodium

Poster 5.6

Composition and In Vitro Study of Alocasia denudata Engler Stem Juice as a Wound Healing Agent

Siti Aisyah Arshad1, Asmah Hamid1, Farah Wahida Ibrahim1 and Mazlyzam Abdul Latif1*
1Biomedical Science Programme, Centre for Health & Applied Sciences, Faculty of Health Sciences, National University of Malaysia, Malaysia

Corresponding author: Dr. Mazlyzam Abdul Latif,


The stem juice of Alocasia denudata Engler is known to possess wound healing properties. This study aimed to analyze the elemental composition and the presence of active compounds in A. denudata stem juice extract and to investigate in vitro wound healing regeneration of A. denudataon human dermal fibroblast cell lines.


The proximate element composition was assessed from freeze-dried sample of A. denudata stem juice extracts using scanning electron microscope (SEM). The active compounds of A. denudata stem juice were analyzed using Gas Chromatography-Mass Spectrometry (GCMS) and Liquid Chromatography-Mass Spectrometry (LCMS). The cytotoxicity of the juice was determined using MTT assay and the cell migration rate was determined by using scratch assay.


Elemental screening showed the presence of oxygen, carbon, potassium, chlorine and sodium and GCMS analysis of the aqueous extract showed the presence of 9-octadecanamide (Z)-, octadecanoic acid-methyl ester, benzyl methyl disulphide as the major components. LCMS analysis revealed the presence of propanoic acid 2-hydroxy-, methyl ester as the major components. All the identified components listed are reported to be conducive for wound healing process. A. denudata extract was proven not cytotoxic to the cells and the highest cell viability was obtained at 62.5 μg/ml. The cells migration towards the wound area also increased compared to the control which shows the highest migration rate at 12-hour. The migration rate was 18 % higher compared to the control suggesting its potential in enhancing wound healing processes.


A. denudata stem juice extract has the potential to be developed as a wound healing agent.

Keywords: Alocasia denudata Engler, Wound Healing, fibroblast, active compounds, cell migration

Poster 5.7

Pharmacokinetic modeling of single dose Kratom (mitragynine) in rats

Yuvenesan Jagabalan1, Hadzliana Zainal1*, Ammar Imad Hazim Al Ganaby2, Vikneswaran Murugaiyah1 and Surash Ramanathan2
1School of Pharmaceutical Sciences, University of Science Malaysia, Malaysia
2Center for drug research, University of Science, Malaysia, Malaysia
Corresponding author: Hadzliana Zainal,


Mitragynine the principal alkaloid of M. speciosa leaf has been reported to be responsible for most of its pharmacological effects. Earlier studies have reported the pharmacokinetic properties of mitragynine (MG) following single oral dose administration. However, the selection of MG doses employed in these studies was derived empirically without proper scientific basis. In view of this, the present study was undertaken to determine MG dose-proportionality properties after repeated oral administration in rats.


In this study, eighteen rats were randomly assigned into three experimental groups (n = 6, each). The first group rats were orally treated with a single dose of 20 mg/kg MG. The second group was given a single oral dose of 40 mg/kg MG whereas the third group of rats received the 80 mg/kg MG dose orally. Blood was collected from the tail vein before the dosing and thereafter at specific time-points after drug administration. Blood samples were centrifuged and separated plasma samples were frozen prior to analysis using HPLC.


The pharmacokinetics was best described by two-compartment distribution models with first order absorption. Estimated typical clearance (CL/F) value was 2.21 L.h-1 , absorption rate (Ka) of of 0.82 h-1 and volume of distribution (Vd) of 30.8 L. The stability of the model and predicted performance were confirmed by bootstrap method and numerical predictive check.


Based on the single dosing experimental rat data, the model provides a useful tool to quantify the pharmacokinetic parameters to propose an optimal dosing regimen in rats. Subsequently, the pharmacokinetic parameter can be modeled to the pharmacodynamics of MG for extrapolation into human use.

Keywords: mitragynine, single dosing study, Two-compartment, Pharmacodynamic (PD), pharmacokinetic

Poster 5.8

In vitro anti-toxoplasmal activities of ethanolic extracts from Andrographis paniculata and Tinospora crispa against Toxoplasma gondii parasite

Sharif A. Abdullahi1, 2, Roslaini Abd Majid1*, Ngah Z. Unyah1, Norshariza Nordin3, Rusliza Basir4, Mohammed N. Wana1, 5, Ashraf A. Alapid1, 6, Hassan Yahaya1, 7 and Tijjani Mustatapha1, 8
1Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia
2Department of Medical Microbiology, Faculty of Clinical Sciences, Bayero University Kano, Nigeria
3Department of Biomedical Sciences, Faculty of Medicine and Health sciences, University of Putra Malaysia, Malaysia
4Department of Human Anatomy, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia
5Department of Biological Sciences, Abubakar Tafawa Balewa University, Nigeria
6Department of Zoology, Faculty of Science, University of Gharyan, Libya
7Department of Medical Laboratory Sciences, Faculty of Allied Health Sciences, Bayero University Kano, Nigeria
8Department of Biological Sciences, Faculty of Science, Yobe State University, Nigeria

Corresponding author: Dr. Roslaini Abd Majid,


Toxoplasma gondii is an intracellular protozoan parasite and one of the infective causes of high morbidity and mortality rates among immunocompromised patients. Drug agents used against toxoplasmosis are associated with treatment failures, attributed to host drug intolerance with severe side effects and possible development of resistance. The need for new treatment modality is prominent especially that there is no available human vaccine against this infection. In this study, we evaluated the effects of A. paniculata and T. crispa extracts on T. gondii in Vero cells.


Ethanolic extracts were obtained through maceration and underwent phytochemical screening. MTT assay was conducted for both cytotoxicity and anti-parasitic activities of both plants. Vero cells were infected and treated with extracts after 4h post infection. We used microscopy to assess parasite cell invasion and intracellular replication in relation to negative control after 24h and 48h. We used clindamycin as a positive control.


The MTT assay revealed that both plants and clindamycin are safe to host cells with IC50 of 142.4, 179±1.5 and 116±2.3 µg/ml for A. paniculata, T. crispa, and clindamycin respectively. Extracts also showed good anti-parasitic activities against T. gondii withIC50 (A. paniculata = 4.36µg/ml, T. crispa = 6.31µg/ml in comparison with clindamycin (8.33±1.04µg/mL). After 24 and 48h of treatment 4h post-infection, the ethanolic extracts were able to cause >60% and > 50% reduction in infection index and intracellular proliferation as compared to clindamycin with >30% in relation to negative control (P<0.001). After 48h, ethanolic extracts caused >70% and >85% reduction of infection index and intracellular proliferation as compared to clindamycin, >25% in relation to negative control (P<0.001).


Our study showed that the ethanolic extracts of both plants contained promising drug candidates effective against T. gondii and safe to the host cells and can possibly be used in future to develop potent anti-toxoplasma compound.

Keywords: Toxoplasma gondii, A. paniculata, T. crispa, Anti-parasitic, Cytotoxicity, cell invasion, Intracellular proliferation, Vero Cells

Poster 5.9

Synergistic effects of gelam honey and fluorouracil (5-FU) on human colorectal cancer HT-29 cells

Syed Ahmad Tajudin Tuan Johari1*, Fatimah Hashim1, wan iryani wan ismail1 and Abdul Manaf Ali2
1Universiti Malaysia Terengganu, Malaysia
2Sultan Zainal Abidin University, Malaysia

Corresponding author: Mr. Syed Ahmad Tajudin Tuan Johari,


Gelam or Melaleuca Honey (GH) is the honey of Apis mellifera from the forest of Melalueca spp in Malaysia. Though it is widely being studied but combinatorial effects with drugs is still lacking. Thus, this study was carried out to determine combinatorial effects of Gelam honey and 5-FU on human colorectal cancer HT-29 cells.


In this study, MTT assay was done to determine the cytotoxic dose. Cells membrane changes was observed in AO/PI assay. Early event of apoptosis detection were done by annexin-v FITC and mitochondria membrane potential. TUNEL assay was performed to examine the fragmented DNA.


The results for MTT assay showed that the IC50 for gelam honey and 5-FU alone were 36.2 mg/ml and 15.5 µg/ml. Meanwhile, the IC50 of combination treatment was 13.2 mg/ml for gelam honey. The combination index was < 1 suggesting the synergism. Early event of apoptosis was occurred in all treatment groups but with different percentage. Combinatorial treatment in apoptosis detection assay showed the highest percentage of apoptotic cells among other treatment groups.


In conclusion, combinatorial treatment of gelam honey and 5-FU may be an effective chemotherapy strategy in fighting colon cancer cells.

Keywords: Gelam honey, Fluorouracil, Apoptosis, cell dealth, Synergistic

Poster 5.10

Anti-angiogenic effect of Ardisia crispa root hexane extract mediated via its angiogenic signalling cascades

Roslida Abd Hamid1*, Lim Wen Jun1 and Chan Pit Foong1
1Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia

Corresponding author:  Dr. Roslida Abd Hamid,


Angiogenesis, defined as the neovascularization from the pre-existing vasculature which involves the endothelial cells proliferation, migration and differentiation, is a vital process that occurs throughout different life stages. However, excessive angiogenesis leads to various diseases, including malignancy and non-neoplastic diseases. Ardisia crispa Thunb A.DC (Myrsinaceae), locally known as “hen’s eyes” in Malaysia, is a local medicinal plant with anti-inflammatory and anti-tumor promoting properties. This study aims to investigate the anti-angiogenic effect of the hexane extract of the plant’s root (ACRH) and its isolated benzoquinonoid (AC2) and its potential pathway(s).


Various angiogenesis assays in vitro against HUVECs, including zebra fish in vivo model were used in this study, followed by multiplex immunoassay in determining the pathway(s) involved.


IC50 for ACRH and AC2 after 24 hours were reported to be 2.49 ± 0.036 and 1.35 ± 0.046µg/mL respectively. ACRH and AC2 showed significant inhibition in a concentration dependent manner (0.1, 1.0 and 10.0µg/mL) in comparison with the negative control (p<0.05) via cell migration, invasion and tube formation assays. In gelatin zymography, pro-MMP2 expression was reduced significantly in HUVEC after treated with ACRH and AC2 in a concentration dependent manner (p<0.05). Angiopoietin-2 (Ang-2), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF)-C and VEGF-D expression were reduced significantly (p<0.05) in a concentration dependent manner, whilst fibroblast growth factor (FGF)-1, FGF-2, Follistatin and heparin binding epidermal growth factor (HB-EGF) were significantly reduced at higher concentration upon ACRH and AC2 treatments, respectively via magnetic bead-based immunoassay. ACRH at 5ug/mL showed significant reduction of intersegmental vessels (ISV) sprouts whilst AC2 showed significant ISV development inhibition at 12.5ug/mL in zebra fish assay.


The present study evidenced the anti-angiogenic effect of ACRH and AC2 through both in vitro and in vivo assays which mediated via angiogenic signaling cascades.

Keywords: Ardisia crispa, Angiogenesis, HUVEC, cell invasion, zebra fish model

Poster 5.11

The effect of administration of an equal dose of different classes of dietary chemicals on heme oxygenase-1 gene expression in mice liver

Azman Abdullah1*, Nadia S. Alrawaiq1 and Ahmed Elbadri2
1Department of Pharmacology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Malaysia
2University of Tripoli, Libya
Corresponding author:  Dr. Azman Abdullah,


Heme oxygenase-1 (HO-1) is an enzyme that possesses anti-oxidant, anti-inflammatory and cytoprotective functions. lnduction of HO-1 occurs as an adaptive and beneficial response to various injurious stimuli such as oxidative stress. This study is aimed at monitoring the effects of equal doses (50 mg/kg) of sulforaphane, curcumin, quercetin, indole-3-carbinol and butylated hydroxyanisole on the expression levels of HO-1 gene.


48 adult male ICR white mice (25–30 g) were divided into 8 groups: normal control group (n=6), sulforaphane treated group (n=6), quercetin treated group (n=6), curcumin treated group (n=6), butylated hydroxyanisole treated group (n=6), indole-3-carbinol treated group (n=6), vehicle 1 control group (n=6) and vehicle 2 control group (n=6). All chemicals were administered intraperitoneally at a dose of 50 mg/kg for 14 days. Vehicle 1 (DMSO, Tween 20 and normal saline at a ratio of 0.05:0.1:0.85) was used to dissolve sulforaphane, quercetin and curcumin. Vehicle 2 (corn oil) was used to dissolve indole-3-carbinol and butylated hydroxyanisole. At day 15, the animals were sacrificed and their livers were isolated. From the liver, total RNA was extracted, reverse transcribed and subjected to quantitative real‐time PCR to detect HO-1 gene expression. Agarose gel electrophoresis was also performed to verify the specificity of the amplification.


Our results showed that there were 4.6, 3.6, 3.6, 3.3 and 3-fold significant increases in HO-1 gene expression levels after treatment with 50 mg/kg of sulforaphane, indole-3-carbinol, butylated hydroxyanisole, curcumin and quercetin respectively, compared to controls (p<0.05).


At a dose of 50mg/kg, sulforaphane administration for 14 days resulted in the highest induction of HO-1 gene expression level in mice liver, and quercetin the lowest.

Keywords: Heme Oxygenase-1, Sulforaphane, Quercetin, Curcumin, Indole-3-carbinol, Butylated Hydroxyanisole, Gene Expression

Poster 5.12

Quantitative Estimation of Residual Solvents in anti-diabetic tablets manufactured by Multinational and National companies in Pakistan

Abida Latif1*, Rukhsana Anwar1 and Asif Raza1
1University of the Punjab, Pakistan

Corresponding author: Prof. Abida Latif,


Residual solvents (RS) are used in pharmaceutical products manufacturing processes from synthesis of active pharmaceutical ingredients/ excipients to the final formulation. RS remain in the final products in the form of volatile impurities can be toxic to health.


An efficient and sensitive headspace gas chromatography (HS-GC) method was developed and validated for determination of twelve RS in thirteen coated anti-diabetes tablets. In order to increase the method sensitivity and efficiency, dimethyl sulfoxide was utilized as diluent owing to its high dissolving capacity for drug substance, stability and boiling point. Headspace auto-sampler was set to multiple extraction mode with 80°C, 90°C and 105°C as oven, loop and transfer line temperatures, respectively. Separation was achieved on HP- Innowax polyethylene glycol gas chromatographic column (30 m × 250 µm × 0.25 µm) using nitrogen as a carrier gas. GC inlet temperature was set at 140°C with split ratio 10:1 and 250°C flame ionization detector temperature.


The method showed good linearity (r2≥ 0.9990) in the range of 50-1000 μg/mL with 5μg/mL as limit of detection. Accuracy and precision at three different concentrations was 90-113% and less than 15% respectively. All tested tablets contained methylene chloride, methanol, n-hexane, toluene and cyclohexane within permissible exposure limits. But in two products (15% of total products tested), the amount of RS were approaching upper limits.


Daily administration of tablets with high RS amount can lead to severe health hazards. It is strongly recommended to include RS analysis in regular pharmaceutical quality control testing protocols.

Keywords: Headspace gas chromatography, Residual solvent, Anti-diabetes tablets, multinational, national

Poster 5.13

Analytical and Biological Studies of Seed Extracts of Daucus carota L. (Black Carrot)

Abida Latif1*
1University of the Punjab, Pakistan

Corresponding author:  Dr. Abida Latif,


 Natural medicinal plants are considered an important approach in search of new therapeutic agents. Even today they were used for the treatment of numerous ailments due to their cost-effective properties, less side effects and high efficacy. Daucus carota L. subsp. sativus (Hoffm.) Arcang. var. vavilovii Mazk. (Family: Apiaceae), is locally known as Kali gajar in Pakistan. Literature has shown that therapeutically important phytochemicals present which strengthen it a suitable candidate for further research. The present study aimed to standardize the seed powder, extracts and then evaluate for in vitro antioxidant, in vitro and in vivo anti-inflammatory activities.


n-hexane, chloroform and methanol extracts were prepared by hot extraction method. The extracts were screened for their antioxidant potential using different in vitro models such as DPPH, FRAP, H2O2, Phosphomolybdenum assay. The in vitro anti-inflammatory activity was carried out by inhibition of protein denaturation method taking diclofenac sodium as standard. The in vivo anti-inflammatory activity was executed using carrageenan induced rat paw edema taking aspirin as standard.


 The proximate analysis revealed that total ash, moisture content present within the stated limits, higher amount of fats (49.1 ± 0.23) than proteins (13.3 ± 0.30) and carbohydrates (31.917 ± 0.50. The minerals detected in mg/g were iron (293), manganese (80 ), zinc (59.5), copper (23.5), nickel (20), lead (6.0) and chromium (5.0). The antioxidant studies revealed that all extracts had significant antioxidant potential but methanol extract exhibited the highest percentage. The antioxidant activity (%) of methanol extract (DPPH, 97; FRAP, 95; H2O2, 89; phosphomolybdenum assay, 67). Anti- inflammatory analysis showed that at higher doses (200 and 400 mg/kg) methanol extract had significant anti-inflammatory activity.


The ability of the extract to scavenge different free radicals indicated that it may be a useful therapeutic agent for treating radical related pathological damage as in rheumatoid arthritis, acute and chronic neurodegenerative ailments, diabetes and inflammatory bowel disease in future.

Keywords: Daucus carota L., seeds, DPPH,, antioxidant activity, anti-inflammatory activity, Inflammation

Poster 5.14

Peroxisome Proliferator-Activated Receptor (Ppar)-ϒ Agonist, Irbesartan and Adiponectin Ameliorate Blood Pressure, Renal Excretory Function and Vasoactive Responses to Adrenergic Agonists and Angiotensin IIin Spontaneously Hypertensive Rats

Sheryar Afzal1*, Munavvar Z. Abdul Sattar1, Javed Qureshi1 and Venkata Sathya S. Raju1
1Faculty of Pharmacy, Mahsa University, Malaysia

Corresponding author: Dr. Sheryar Afzal,


Adiponectin (Adp) possesses vasodilatory effect through nitric oxide release. Irbesartan (Irb) possesses natriuretic properties, causes enhanced plasma adiponectin concentration. This study explores the effect of irbesartan alone and in combination with adiponectin on blood pressure, renal haemodynamicparameters and vasoactive responses to angiotensin II (Ang II) and adrenergic agonists in spontaneously hypertensive rats (SHRs).


Irbesartan treated groups (SHR+Irb) and SHR+Irb+Adp groups received irbesartan (30mg/kg) orally for 4 weeks, whereas SHR+Adp and SHR+Irb+Adp groups were administered adiponectin intra-peritoneally at 2.5µg/kg/day, for last one week. A group of Wistar- Kyoto (WKY) rats served as normotensive control,(n=6). Metabolic data and plasma samples were collected at day 0, 21, 28 and were analyzed for creatinine and adiponectin concentrations.


In acute study, the renal vasoconstrictor actions of Ang II, noradrenaline (NA), phenylephrine (PE) and methoxamine (ME) were determined, thereafter animals were anaesthetized. Data, mean±SEM were analysed using ANOVA with significance at P<0.05. SHR control rats had higher mean arterial blood pressure (132.0 ±7 vs. 98.0±2 mmHg), lower plasma and urinary adiponectin, creatinine clearance & urinary sodium excretion compared to WKY. Responses to NA, PE, ME and Ang II were higher than WKY (SHR vs. WKY—50±4 vs. 38±3%, 60±3 vs. 49±2%, 48±4 vs. 33±3%, 41±3 vs. 55±3%). Irbesartan had greater responses to NA and PE while adiponectin had lesser responses to NA, PE, ME and Ang II compared to SHR (SHR+Irb, SHR+Adp vs. SHR, 63±3, 24±2 vs. 50±4; 72±3, 34±3 vs. 60±3; 50±3, 48±4 vs. 21±3; 30±2, 25±4 vs. 55±3). SHR+Irb+Adp further decreased responses to Ang II compared to SHR+Adp and SHR+Irb (18±4, 30±4, 25±2), (P <0.05).


Data indicates that an interactive relationship exists between Ppar-ϒ, alpha adrenoceptors and Ang II in renal vasculature of SHRs. Adiponectin with irbesartan ameliorates renal cortical blood perfusion and normalized kidney function compared with either compound alone.

Keywords: Irbesartan, Adiponectin, Angiotensin II., agonists, adrenergic

Poster 5.15

Impact of Hyperuricemia on the Progression of Renal Disease among Patient in a tertiary renal center

Mohammad Zulkarnain Bidin1, Christopher T. Lim1, 2*, Anim Md Shah1, 2, Johnson Stanslas1 and Bak Leong Goh2
1Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia
2Serdang Hospital, Malaysia

Corresponding author: Dr. Christopher T. Lim,


Chronic Kidney Disease (CKD) is associated with the estimation of 188 million cases of catastrophic health expenditure in the low and middle-income countries, thus seriously affecting a country health and productivity. Studies have reported association of hyperuricemia and other comorbidities with the progression of CKD, but their conclusions often contradict with one another. We analyzed the association between the serum uric acid, etiologies of CKD, co-morbidities, laboratory parameters with the rate of decline in eGFR.


This is an ambispective study conducted in the nephrology clinic, Serdang Hospital. A preliminary 100 out of an intended cohort of 400 patients that were referred to nephrology were followed up for a minimum of 6 months from 2007 until June 2018. Data was collected via proforma sheet and extracted from the Hospital eHIS system. This research is registered with NMRR (NMRR-18-866-41231). The Kaplan-Meier survival curve were used to analyze the renal outcomes, namely the decrease of 50% rate of eGFR decline, doubling serum creatinine and need for renal replacement therapy (RRT) with regards to the effects of hyperuricemia. Cox regression model was used to assess the influence of hyperuricemia, age, gender, weight, height, smoking and laboratory parameters on the renal outcomes. A P-value < 0.05 was considered as significant.


There is no association of renal outcome (decline in eGFR (p = 0.427), doubling serum creatinine (p = 0.117), RRT initiation (p = 0.332) with the presence of hyperuricemia. Moreover, multivariate cox regression analysis did not show any significant association with the renal outcome (50% reduction in GFR (p= 0.855, HR: 1.073), doubling serum creatinine (p= 0.095, HR: 0.354), and initiation of RRT (p= 0.087, HR: 0.548) with hyperuricemia.


Hyperuricemia is not associated with worsening of renal function in our interim analysis. A larger cohort study which is on-going will be able to provide more information on the relationship of uric acid with renal function.

Keywords: Hyperuricemia, CKD, EGFR, serum creatinine., RRT – renal replacement therapy

Poster 5.16

Investigation of anti hypertensive activity of standardized Colebrookea oppositifolia methanolic extract and elucidation of possible mechanism of action

Rukhsana Anwar1*, Muhammad U. Khalil1 and Abida Latif1
1University of the Punjab, Pakistan

Corresponding author:  Dr. Rukhsana Anwar,


Colebrookea oppositifolia belongs to family Lamiaceae and locally known as ‘bansa siah’ in Pakistan. Traditionally, it is used as analgesic, anti-inflammatory, to treat asthma and epilepsy. No study is available for its anti hypertensive activity. Following study was designed to investigate in vivo acute and sub-acute anti hypertensive effect of methanolic extract of Colebrookea oppositifolia leaves in hypertension-induced rats. Possible mechanism of anti hypertensive action was also elucidated by estimating angiotensin conversing enzyme activity.


Hypertension was induced by 10% glucose administration orally for 21 days. In vivo acute anti hypertensive effect was investigated by two doses (250 & 500mg/kg) of methanolic extract as a single dose. Methanolic extract of Colebrookea oppositifolia 250mg/kg administered for 14 days to investigate sub-acute anti hypertensive effect. Captopril (20mg/kg) administered as control group. Rat blood pressure was measured using non-invasive blood pressure (NIBP). The angiotensin converting enzyme activity was assayed by monitoring the rate of production of hippuric acid from the hydrolysis of Hippuryl-L-Histidyl-L-Leucine (HHL).


Ten percent glucose administration increases systolic blood pressure, diastolic blood pressure, mean arterial blood pressure and heart rate significantly. Acute study results showed that test doses of extract significantly (p<0.05) decreased systolic blood pressure 96.75±3.9 & 104±3.8, diastolic blood pressure 73.12±5.2 & 65.37±8.2 mean arterial blood pressure 81±4.1 & 78.25±6.7 and heart rate 370±33.9 & 298.25±19.1 respectively as compared to hypertensive rats. Sub-acute study results showed that the extract significantly decreased systolic blood pressure 97.33±3 diastolic blood pressure by 68.6±4.6 mean arterial blood pressure 78.17±3.7 and heart rate 335.8±27.5 as compared to hypertensive control rats. Moreover, methanolic extract inhibited 45% ACE activity in hypertensive rats as compare to normal control group.


 This research concluded that methanolic extract of Colebrookea oppositifolia leave has potential to reverse the hypertension and its anti hypertensive action may be due to inhibition of ACE.

Keywords: Hypertension, Colebrookea oppositifolia, Noninvasive blood pressure (NIBP), Diastolic blood pressure, Angiotensin – converting enzyme (ACE)

Poster 5.17

Alleviating Effect of Mitragynine on Microsomal UDP-Glucuronosyltransferase Activity and UGT1A6 Expression in STZ-Induced Diabetic Rat

Rukhsana Anwar1, 2*, Sabariah Ismail2, Sharif M. Mansur2 and Tan M. Lan2, 3
1Punjab University College of Pharmacy, University of the Punjab, Pakistan
2Centre for Drug Research, University of Science, Malaysia, Malaysia
3Center for drug research, University of Science, Malaysia, Malaysia

Corresponding author: Dr. Rukhsana Anwar,


Mitragyna speciosa (MS) Korth is a member of the Rubiaceae family, found in Peninsular Malaysia and Thailand. MS leaves are being used for opium withdrawal, as analgesic and to treat diarrhea. Mitragynine (MTG) is a major indole alkaloid of MS leaves. Its analgesic, antipyretic and behavioral effects have been studied. Efficacy and safety are essential parameters during herbal derived drug development. It is only possible by evaluation of types of enzymes involved in its metabolism. Till now, no in vivo study has been done about the effect of mitragynine on UDP-glucuronosyltransferase (UGT) phase II drug metabolizing enzyme activity. Herbal derived compounds modulate the expression of hepatic enzymes that is an important mechanism determines the bioavailability of co-administered drug.


The following research was undertaken to investigate effect of MTG on UGT activity in healthy and diabetic male Sprague Dawley (SD) rats. Diabetes was induced by streptozotocin. For sub-acute in vivo study, 50 mg/ kg mitragynine (single oral daily dose for 14 days) administered to healthy and diabetic rats. Pooled liver microsomes were prepared by differential centrifugation technique. The p-nitrophenol was used as a probe substrate for estimation of UGT activity in rat liver microsomes. Effect of MTG on the UGT1A6 protein expression rat liver microsomes were measured in three independent immunobloting assays. The protein band was quantified by densitometry and the level of control was set at 1.


The sub-acute administration of mitragynine to normal male rats showed significant (p < 0.05) 189% increase in UGT specific activity and diabetic rats showed 172% induction in UGT specific activity as compared to respective control group. The fold change in UGT1A6 protein expression in both groups was more than 2-fold indicating that MTG was a significant UGT1A6 inducer.


Mitragynine significantly induced activity of UGT & expression of UGT1A6 in normal and diabetic rats.

Keywords: UGT, mitragynine, Microsomes, UGT1A6, Immunobloting

Poster 5.18

Formulation and In-Vitro Evaluation of Amoxicillin Mucoadhesive Microcapsules for the Treatment of Helicobacter pylori Infection

Senthil Adimoolam1* and Bolai Paul1
1Mahsa University, Malaysia

Corresponding author:  Dr. Senthil Adimoolam,


Amoxicillin (α-amino-p-hydroxybenzyl-penicillin) is a semi-synthetic, orally absorbed and widely prescribed β-lactam antibiotic. It is widely used for eradication of gastric Helicobacter pylori infection combined with a second antibiotic and an acid‐suppressing agent despite its short elimination half-life of one hour. The purpose of this study was to develop and evaluate amoxicillin loaded HPMC K15M mucoadhesive microcapsules for sustained drug release at the gastric mucosa to prolong residence time of dosage form in stomach to achieve more effective H. pylori eradication.


Amoxicillin mucoadhesive microcapsules were formulated by ion gelation technique by using different mucoadhesive polymers HPMC K15M and sodium alginate with varying concentration. A 32 full factorial design was applied to derive statistical equation, ANOVA analysis, contour plots and 3D response surface plots. The nine formulations were subjected to evaluation of percentage yield, entrapment efficiency, swelling index, mucoadhesion and in vitro drug release.


Among nine formulations (F1 – F9), F9 exhibited a promising drug entrapment efficiency (92.50±0.03%), percentage yield (97.50±0.02%), percentage mucoadhesion (65±0.03 after 8 h) and in vitro drug release more than 14 h. The kinetics of the drug release of microcapsules (F9) showed good fitting with zero order and Korsmeyer-Peppas model followed by a non-Fickian release. The ANOVA analysis shows highly significant effects on depended variable.


Therefore, amoxicillin mucoadhesive microcapsules can be effectively used in the treatment of H. pylori infection.

Keywords: microcapsule, Amoxicillin, mucoadhesion, Entrapment efficiency, Percentage yield

Poster 5.19

Demystifying Aspirin Toxicity Through Pharmacometabonomics

Abubakar Sha’Aban1*, Hadzliana Zainal1, Muhamad Ali S. Abdul Kader2 and Baharudin Ibrahim1
1Discipline of Clinical Pharmacy, School of Pharmaceutical Sciences, 11800 Universiti Sains Malaysia, Malaysia
2Cardiology Department, Hospital Pulau Pinang,, Malaysia

Corresponding author:  Mr. Abubakar Sha’Aban,


Just like many nations, cardiovascular diseases (CVDs) remains the principal cause of death for more than a decade in Malaysia. Among the CVDs, Coronary Artery Disease (CAD) is the leading cause of such mortalities. In 2016, CAD accounted for 13.2% of all recorded deaths (162,201 deaths) in Malaysia. Low Dose Aspirin (LDA) is the cornerstone of secondary prevention in CAD. Despite its established efficacy, it has a prime limitation arising from its gastric toxicity. This may manifest as either dyspepsia, ulcer, and/or bleeding. The reason some patients develop clinically significant toxicities while others develop no or minor toxicities is yet to be demystified. This study aims to personalise LDA therapy by identifying biomarkers for LDA-induced gastric toxicity in CAD patients using Pharmacometabonomics approach.


Patients that satisfy the study inclusion criteria were recruited after informed consent. Urine samples of recruited patients were obtained at baseline (Pre) and after one month (Post) of initiating LDA therapy. The Pre- and Post-LDA urine were analyzed using Proton nuclear magnetic resonance (1H NMR) spectroscopy to identify the biomarkers that may distinguish between those with and without gastric toxicity.


Thirty-eight percent (38%) of the patients that meet up with the inclusion criteria had a history of gastric toxicity. Preliminary results of the 1H NMR pharmacometabolomic analysis show some low molecular weight metabolites that are potential biomarkers for LDA-induced gastric toxicity in CAD patients. The key feature here is that pre-dose urine samples were exploited for the presence of discriminating metabolites that may predict the occurrence of LDA-induced gastric toxicity. Based on this, LDA can safely be avoided in such patients with confirmed presence of such metabolites.


This study clearly demonstrates the gastric toxicity of LDA. Multivariate statistical analysis and validation to indicate the discriminating metabolites that may be linked to the gastric toxicity is ongoing.

Keywords: Pharmacometabonomics, Low dose aspirin, biomarkers, Coronary Artery Disease, Gastric toxicity

Poster 5.20

Chromosomal Mutation in Isolated Hypospadias Patients Using
High-Resolution Melting Curve (HRM)

Darmawati Ayu Indraswari1, Ahmad Zulfa Juniarto2, Andrew Sinclair3, Sultana MH.  Faradz2
1Faculty of Medicine, Diponegoro University, Semarang Indonesia 50275
2Centre for Biomedical Research, Faculty of Medicine, Diponegoro University, Semarang Indonesia 50275
3Murdoch Childrens Research Institute, Victoria Australia 3052


Hypospadias, one of the most common disorders of male genitalia is thought to have an increased trend in prevalence worldwide. An important gene involved in the development of male external genitalia is WNT5A. Besides, the increased frequency in the last 20 years has been thought to be associated with environmental changes especially in the increased risk of exposure to chemical compounds. Currently there was no study using high-resolution melting curve (HRM) for hypospadias. Aim: To identify mutations in hypospadias patients using HRM.


Samples are 17 Indonesian and 27 Pakistani DNA with a clinical diagnosis of hypospadias. HRM was employed, continued with Sanger sequencing if aberrant patterns are found.


Aberrant patterns were found using HRM in two samples and further analysis using Sanger sequencing resulted in translocations of chromosome 8: 143516989-143517080 (92 nt) and 143516981-143517080 (100 nt) in two samples consecutively.


Translocations of chromosome 8 were found in two samples of hypospadias patients.

Keywords: Hypospadias, HRM, WNT5A