Oral 1.1

Identification and development of novel anti-lymphangiogenic compounds as cancer therapeutics

Kazuhide S. Okuda1, Mei Fong Ng1, Norazwana Samat1, Nur Faizah Ruslan1, Pei Jean Tan1* and Vyomesh Patel1

1Cancer Research Malaysia, Malaysia

Corresponding author: Pei-Jean Tan, peijean.tan@cancerresearch.my


Cancer metastasis is the leading cause of death in cancer patients as it accounts for approximately 90% of human cancer death. It is well reported that blood and/or lymphatic vessels, play key roles in cancer metastasis by providing nutrient and conduits for cancer cell dissemination. Consequently, many anti-angiogenic agents are developed and currently used in clinics. Although promising, these are nevertheless associated with side effects. Alternatively, the intimate relationship between cancer metastasis and lymphangiogenesis has stimulated researchers to seek for novel anti-lymphangiogenic drugs as therapeutic options for cancer metastasis. Though there are few anti-lymphangiogenic agents which are currently under clinical evaluation, there are still no FDA approved selective agents designed to prevent lymphatic vessel growth, highlighting an important clinical niche. In this study, we aim to identify novel anti-lymphangiogenic compounds as novel therapeutic agents for metastatic human cancer.


 Zebrafish is an established in vivo model for vascular biology and had contributed in identifying various novel anti-(lymph)angiogenic compounds. Here, we utilized the Tg(lymphatic vessel endothelial hyaluronic receptor 1b (lyve1b):Discosoma sp. Red fluorescent protein 2 (DsRed2))nz101 transgenic that express DsRed2 in all lymphatic/venous endothelial cells, to screen for anti-(lymph)angiogenic compounds from our compound libraries consisted natural and semi-synthetic compounds. The thoracic duct development in zebrafish was quantified using fluorescent microscope to measure lymphatic vessel development.


Approximately 300 compounds were screened and we identified 2 semi synthetic compounds which inhibited thoracic duct development specifically. Subsequent biochemical analysis revealed inhibition of phosphorylated ERK1/2 expression, the down-stream target of vascular endothelial growth factor (VEGF) pathway in human lymphatic endothelial cells upon compound treatments, suggesting anti-lymphangiogenic activity through inhibition of VEGF pathway.


Novel anti-lymphangiogenic compounds can be identified using the zebrafish model and potential therapeutics can be developed against cancer metastasis, by targeting lymphangiogenesis.  

Keywords: cancer, metastasis, zebrafish, angiogenesis and lymphangiogenesis

Oral 1.2

Exploring the Potential of Andrographolide and Its Derivatives as Anti-Pancreatic Cancer Therapeutics: In silico, In vitro and In vivo approaches

Shun Ying Quah1, Kok Lian Ho1, Nizar Abdul Manan1, Sreenivasa R. Sagineedu2, Pran Kishore Deb3 and Johnson Stanslas1*

¹Department of Medicine, ²Department of Pathology, ³Department of Human Anatomy, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
⁴Department of Pharmaceutical Chemistry, International Medical University, Bukit Jalil, Kuala Lumpur, Malaysia
⁵Faculty of Pharmacy, Philadelphia University, Amman, Jordan

Corresponding author: Johnson Stanslas, rcxjs@upm.edu.my/jstanslas@yahoo.co.uk


Approximately 95% of pancreatic ductal adenocarcinoma (PDAC) patients are found having oncogenic K-Ras, which perturbs mitogen-activated protein kinase (MAPK) cascade critical for cellular processes. Our previous analysis identified andrographolide (AGP) derivatives, SRJ09 and SRJ23, disrupted MAPK activation by binding to K-Ras. Chemical modification of SRJ23 produced S8, which selectively inhibited PDAC cell growth. This study aims to evaluate these compounds computationally on K-Ras, and their in vitro and in vivo anticancer activity against PDAC.


Using Glide (Schrödinger, LLC), molecular docking was performed to identify the binding mode and affinity of AGP and its analogues to wild-type and mutant K-Ras. PANC-1 cells harbouring K-RasG12D, the most predominant K-Ras mutation, were used in cell-based and animal studies. Soft-agar colony formation assay was performed to determine the ability of compounds to inhibit anchorage-independent cell growth. GTP-loading of K-Ras and MAPK activation were assessed by immunoblotting. Anti-tumour activity was assessed in athymic nude mice carrying PANC-1 tumour xenografts. Tumour tissues were evaluated histologically by haematoxylin-eosin staining.


S8 bound stronger to K-Ras mutants, specifically K-RasG12D and K-RasG12V (-76.54 kcal/mol and -80.43 kcal/mol, respectively, versus -69.40 kcal/mol in wild-type K-Ras). Contrary to AGP and SRJ09, SRJ23 and S8 anchored into a binding pocket located between two switches of K-RasG12V, via their lactone ring, where the 14-OH group interacted with aspartate-54 through hydrogen-bonding. SRJ23 and S8 comparably inhibited PANC-1 colony formation in a dose-dependent manner. However, S8 produced greater repression in GTP-loading of K-Ras and suppressed Erk phosphorylation in PANC-1 cells. S8 significantly decelerated tumour growth in PANC-1-xenografted nude mice at a dose of 100 mg/kg, and caused a higher degree of tumour necrosis, in comparison to SRJ23 treatment at the same dose.


S8 likely binds to oncogenic K-Ras, leading to inhibition of the aberrant Ras-MAPK signalling and reduction in pancreatic tumourigenesis in vitro and in vivo.  

Keywords: PDAC, oncogenic K-Ras, andrographolide derivatives, MAPK, tumourigenesis

Oral 1.3

Identification of molecular targets of Tris DBA [Tris(dibenzylideneacetone)dipalladium(0)] for cancer therapy

Loukik Arora¹, Gautam Sethil¹*

¹Department of Pharmacology, YLLSoM, National University of Singapore, Singapore

Corresponding author: Gautam Sethi, phcgs@nus.edu.sg


Cancer is a complex disease involving complex genetic and epigenetic heterogeneity making it the second leading cause of death globally after heart disease. While conventional treatments like surgery, radiotherapy, chemotherapy are principal strategies, the focus is shifting to therapies that can target multiple pathways to treat cancers. We hypothesized that Tris DBA, an organometallic compound, might inhibit proliferation and tumor growth, and induce apoptosis in multiple myeloma (MM) and hepatocellular carcinoma (HCC) cells.   


In silico

Cell Works© (USA)- target pathway and subsequenty affected genes and proteins prediction.

In vitro

MTT assay was used to assess cell viability and thereby effect of the compound. Flowcytometric analysis was used to study the effect on cell cycle progression. Western blotting was performed to check for protein expression as well as activation/deactivation upon treatment. Further, wound healing assay and invasion assay were used to study effect on metastatic potential and con-focal microscopy for protein localisation changes.

In vivo

Acute toxicity as well as efficacy studies were carried out using nude mice and orthotopic HCC mouse model respectively.


The in silico results narrowed down a few key pathways including STAT3 as potential targets. Following up on that,our preliminary results have clearly indicated that Tris DBA could substantially inhibit both constitutive and IL-6 inducible STAT3 activation and abrogate proliferation/survival of MM(IC50-2.6µM) and HCC(IC50-3.6µM) cells. In vivo inhibition of tumour growth in HCC orthothotopic mouse model has also been shown at various clinically relevant doses with no indication of acute toxicity in mice.


Tris DBA exhibits a broad spectrum of anticancer effects in both solid and hematological malignancies. Further elucidation of the molecular mechanism(s) underlying STAT3 inhibitory effects of the drug is underway using chemi proteomic and phospho proteomic approach.

Keywords: Tris DBA, STAT3, signaling pathways, apoptosis

Oral 1.4

Antileukaemic effect of palladium nanoparticles mediated by white tea (Camellia sinensis) extract in WEHI-3B cell-induced murine leukaemic model

Hemn Othman¹,²*, Heshu Rahman²,³, Mohammed Hasan⁴

¹Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
²College of Veterinary Medicine, University of Sulaimani, Sulaimani, Kurdistan Region, Republic of Iraq
³Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
⁴Faculty of Veterinary Medicine, University of Al Basra, 61004, Republic of Iraq

Corresponding author: Hemn Othman, hemn.othman@upm.edu.my


Cancer nanotherapy is progressing rapidly with the introduction of many innovative nanocarriers using herbal extract as the medium. Palladium nanoparticles (Pd@W.tea NPs), in particular, were reported to exert antiproliferative effects against human leukaemic cells (MOLT-4) through induction of apoptosis and G2/M cell-cycle arrest. As part of the effort in confirming the promising role of Pd@W.tea NPs for treatment against cancers, both in vitro and in vivo antileukaemic effects of Pd@W.tea NPs against WEHI-3B cell-induced murine leukaemia were investigated.


Palladium nanoparticles, which was developed by green synthesis using white tea (Camellia sinensis) extract, was characterised by zetasizer, UV–vis spectroscopy, X-ray diffractometry, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The in vitro antileukaemic effect was determined using MTT, Annexin V and cell cycle assays.  The in vivo effects of Pd@W.tea NPs in treated leukaemic mice was evaluated through morphological study, histopathological changes and Immunohistochemistry (IHC).


The results from MTT, Annexin V, and cell cycle assays showed that Pd@W.tea NPs reduced the growth of WEHI-3B leukaemic cells at IC50 of 7.55 µg/mL. Furthermore, outcomes of histopathology and IHC analyses found that the number of leukaemic cells in the spleen of leukaemic mice were significantly lowered to less than 50% after 28 days of oral treatment with of Pd@W.tea NPs at 50 and 100 mg/kg. In addition, western blotting and reverse-transcription quantitative polymerase chain reaction (RT-PCR) assays confirmed the antileukaemic effects of Pd@W.tea NPs through significant (p<0.05) up-regulation of Bax and Cytochrome C genes, with down regulation of Bcl-2 gene.


Pd@W.tea NPs, which induced an intrinsic apoptotic pathway in murine leukemia, could be potentially developed into an anticancer agent.

Keywords: Natural products, green synthesis, nanoparticles, antileukaemic therapy, apoptotic pathway.

Oral 1.5

The Effect of Carica pubescens Leaf Extract on Haematological Profile of Azoxymethane-Induced Colon Cancer Rats

Ainun Rahmasari Gumay¹*, Saekhol Bakri², Eka Meyliana Sugeng³, Maharani Shofa Yudina⁴, Faradilla Nadya Bleizensky³, Indah Saraswati⁴, Muflihatul Muniroh¹, Yosef Purwoko¹, Hardian¹

¹Department of Physiology, Faculty of Medicine, Diponegoro University, Semarang, Indonesia
²Department of Public Health, Faculty of Medicine, Diponegoro University, Indonesia
³Bachelor Student, Faculty of Medicine, Diponegoro University, Semarang, Indonesia
⁴Department of Chemistry, Faculty of Medicine, Diponegoro University, Semarang, Indonesia

Corresponding author: Ainun Rahmasari Gumay. Email: ainungumay@fk.undip.ac.id


Neuroinflammatory and apoptosis mechanisms play an important role in the pathogenesis of colorectal cancer. Carica pubescens (CP) has potential anti-inflammatory, antioxidant and anticancer effects. This study aims to determine the effect of CP leaf extract on haematological profile of azoxymethane (AOM)-induced colon cancer rats.


Sprague Dawley rats (25 male, aged 5-7 weeks) were divided into 5 groups. The AOM groups were induced intraperitoneally by injection of AOM (10 mg/kgBW) once weekly for 2 weeks. CP-100, CP-200, and CP-400 groups were induced by AOM and orally administered by 100, 200, and 400 mg/kgBW CP leaf extract once daily for 2 weeks. The normal control (NC) group was given saline. The haematological profile that was examined includes white blood cell (WBC), lymphocyte, and neutrophil count. One-way Anova and post hoc LSD were used for statistical analysis.


The WBC count of CP-100(4380±715,5/μL) was significantly lower than AOM (7000±2065,2/μL; p=0,002). The lymphocyte count of CP-100 (3260±746,9/μL) was also significantly lower than AOM (5460±1647,1/μL; p=0,001). The neutrophil counts in CP-100 (940±554,9/μL; p=0,005), CP-200 (1220±342,1/μL; p=0,001), and CP-400 (1240±680,4/μL; p=0,008) were significantly lower than the AOM group (2040±270,2/μL)


Carica pubescens leaf extract seem to reduce the number of WBC, lymphocytes, and neutrophil counts in azoxymethane-induced colon cancer rats.

Keywords: Carica pubescens, colon cancer, azoxymethane

Oral 1.6

Chemopreventive and antioxidant potential effects of oleuropein on two-stage skin carcinogenesis mouse model

Dayang Noor Suzliana John¹, Omchit Surien¹, Zariyantey Abdul Hamid¹, Siti Fathiah Masre¹*

¹Biomedical Science Programme, Centre of Health & Applied Sciences, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.

Corresponding author: Siti Fathiah Masre, sitifathiah@ukm.edu.my


Oleuropein is a phenylethanoid, a form of a phenolic compound which found in olive leaf. It is known to have numerous pharmacological roles such as antioxidant, antibacterial, anti-inflammatory and anti-tumour. This study was carried out to investigate the effect of oleuropein on the two-stage skin carcinogenesis mouse model including the initiation and promotion stages.


Skin tumours were initiated by topical application of 7,12-dimethylbenz[a]anthracene (DMBA: 200nmol) and promoted by 12-O-tetradecanoylphorbol-13-acetate (TPA: 20nmol) on the shaved dorsal area of mice.  Total of 30 female ICR mice was randomly divided into five groups (n=6 per group). Group I was treated with DMBA and TPA, group II as a negative control was treated with acetone (70%) alone, group III to V are treatment groups (10mg/kg): oleuropein pre-initiated group (Group III), oleuropein post-initiated group (Group IV) and oleuropein pre- and post-initiated group (Group V).


After 16 weeks, group III showed a significant reduction in the percentage of mouse bearing tumour (p<0.05) and the mean number of tumour per mouse (p<0.05) as compared to group I. Based on histopathological analysis, group III prevented skin carcinogenesis with mild epidermal hyperplasia formation even after 16 weeks of DMBA/TPA induction. Moreover, the level of lipid peroxidation (MDA) was significantly reduced in group III compared to group I (p<0.05). The level of antioxidant superoxide dismutase (SOD) was significantly increased in group III (p<0.05), while the level of glutathione (GSH) was significantly higher in Group V (p<0.05) compared to group I.


These findings indicate that oleuropein may act as a potent chemopreventive agent at the pre-initiation stage against the development of mouse skin carcinogenesis through its antioxidant actions.

Keywords: oleuropein, skin cancer, chemoprevention, antioxidant, carcinogenesis

Oral 1.7

Phytochemical analysis and augmentation of pulmonary adenocarcinoma in BALB/c mice model treated with flaxseed oil

Heshu Rahman¹,²*, Hawsar Mohamad², Hardi Fattah², Hemn Othman²,³, Kawa Amin⁴, Rasedee Abdullah³,*

¹Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
²College of Veterinary Medicine, University of Sulaimani, Sulaimani, Republic of Iraq
³Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
⁴College of Medicine, University of Sulaimani, Sulaimani, Republic of Iraq

Corresponding authors: Heshu Sulaiman Rahman, heshusr@upm.edu.my and Rasedee Abdullah, rasedee@upm.edu.my


Lung cancer is a highly invasive and metastatic tumor that causes more than 1.4 million deaths annually. Thus, the current study was aimed to evaluate the anti-tumor and anti-metastasis effects of flaxseed oil (FSO) in a mice model of pulmonary adenocarcinoma.


The total phenol and flavonoid contents in the FSO were determined using Folin–Ciocalteau and the aluminum chloride colorimetric assays. Later on, 30 male Balb/C mice were randomly allotted into 5 experimental groups of 6 animals each. Lung cancer was induced in all groups except group 1 using nitrosamine 4-(methyl-nitrosamino)-l-(3-pyridyl)-1-butanone (NNK). Lung tumour-induced BALB/c mice were fed with either 150 or 300 mg/kg body weight of FSO, or Erlotinib (50 mg/kg body weight). After 28 days, animals were sacrificed, and biochemical analysis was conducted on collected blood samples to detect the changes of white blood cell (WBC) and lymphocyte counts. On the other hand, collected lung tissues were stained with Hematoxylin and Eosin (H&E) to detect the histopathological abnormalities, whereas immunohistochemistry (IHC) was performed to observe the biomarkers expression related to cancer and inflammation in the lung tissues.


Using phytochemical analysis, we found that the FSO is rich in tannin and gallic acid, which are the main phenolic compounds in the oil. Regarding the animal study, the FSO significantly (P < 0.05) increased WBC and lymphocyte counts. After IHC staining, we realized that the FSO reduced the epidermal growth factor receptor (EGFR), lung adenocarcinoma biomarker, suppressed the cyclooxygenase-2 (COX-2), inflammatory biomarker and enhanced the phosphatase and tensin homolog (PTEN), a tumor suppressor gene consequently.


We found that the FSO showed promising potential as a complementary therapeutic dietary supplement which was effective in suppressing lung adenocarcinoma.

Keywords: Flaxseed oil, Lung adenocarcinoma, antitumor, antimetastatic, epidermal growth factor receptor.

Oral 1.8

Enhanced anticancer effect of aerosolized nanoemulsion system containing docetaxel and curcumin in lung carcinoma cells via synergistic approach

Azren Aida Asmawi¹,², Norazlinaliza Salim¹,², Mas Jaffri Masarudin³ and Mohd Basyaruddin Abdul Rahman¹,²*

¹Integrated Chemical BioPhysics Research, Faculty of Science, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
²Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
³Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia

Corresponding author: Mohd Basyaruddin Abdul Rahman, basya@upm.edu.my


Lung cancer is the most common cause of cancer-related deaths worldwide. Synergistic anticancer effect of docetaxel and curcumin may emerge as an attractive therapeutic candidate in the lung cancer treatment. However, lacks of their optimal bioavailability due to poor solubility, low stability and high toxicity have limited their clinical success. Hence, attention has been focused on the use of inhalable nanoemulsion system for localized drug delivery to overcome the challenge.


In this study, nanoemulsion formulations containing biocompatible lipid excipients were prepared by ultrasonic-assisted emulsification technique. The efficiency and feasibility of the formulations were characterized physicochemically and aerodynamically with an emphasis on the required criteria for pulmonary delivery. In addition, investigations on the anticancer effect in A549 lung carcinoma cells were also evaluated.


The formulated docetaxel and curcumin-loaded nanoemulsion system was observed to be in nano-sized, homogenously dispersed along with the pH, osmolality and viscosity values met the ideal requirement for pulmonary application. Moreover, the formulation exhibited sustained drug release and excellent physical stability against extreme conditions. The aerosolized nanoemulsion system was able to be deposited on the deep lung region with aerodynamic size less than 5 µm. Interestingly, cytotoxicity assay of the combined drugs formulation showed higher cytotoxic effect in A549 cells with reduced cytoxicity in normal lung cells (MRC-5) compared to single drug formulation.


Overall, this synergistic drugs combination using nanoemulsion formulation for inhalation mode has significant promise in lung cancer therapy.

Keywords: nanoemulsion, docetaxel, curcumin, pulmonary delivery and lung cancer

Oral 1.9

Photocytotoxic Effects of Strobilanthes crispus Extracts Towards Human Skin Squamous Carcinoma

Shi Yun Lim¹, Chui Fung Loke¹*, Sheri-Ann Tan¹, Tze Ven Poh², Wan Zuhainis Saad³, Saila Ismail³, Khatijah Mohamad Yusoff ³

¹Department of Bioscience, Faculty of Applied Sciences, Tunku Abdul Rahman University College, 53300 Setapak, Kuala Lumpur, Malaysia
²Department of Computer Science and Embedded Systems, Faculty of Computing and Information Technology, Tunku Abdul Rahman University College, 53300 Setapak, Kuala Lumpur, Malaysia
³Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Selangor, Malaysia

Corresponding author: Chui Fung Loke, lokecf@tarc.edu.my


The killing mechanism by potential photosensitizing chlorophyll-metabolites from Strobilanthes crispus in human skin squamous carcinoma (A431) cells was investigated. The investigation focused on the mechanisms of cell death by the mitochondrial pathway involving death receptors/caspase activation; photodamage of Bcl-2 family in photodynamic therapy (PDT) response, as well as the photosensitivity efficacy of the plant-extracts.


Cell viability and reactive oxygen species (ROS) generation after PDT were measured using MTT and DCFH-DA assays respectively. Specific morphological and biochemical changes including cell shrinkage, membrane blebbing, DNA fragmentation, and regulation of anti-/pro-apoptotic proteins such as Bcl-2, Bax and caspase-3 were examined.


No dark toxicity was observed at concentrations up to 500 μg/ mL (for crude extract) and 5.0 μg/mL (for sub-fraction containing pheophorbidea) in A431 cells. Surprisingly, upon 10 min PDT, apoptotic morphologies appeared in S. crispus-treated A431 at mild concentration ≤ 2 μg/ mL of sub-fraction, while autophagic morphologies occurred in high concentration of ≥ 4 μg/ mL sub-fraction-treated A431. The reduction in cellular viability dramatically charting an IC50 of 9.61 μg/mL for crude extract (containing 0.004 μg/mL of pheophorbide-a) and 2.20 μg/mL for sub-fraction (containing 0.001 μg/mL of pheophorbide-a) respectively. However, IC50 of pheophorbide-a (> 90% purity) on A431 upon PDT was 23-fold (0.085 μg/mL) lower than sub-fraction of S. crispus leave extract. ROS increased remarkably with the descending of A431 cell viability upon PDT. Fragmented DNA was observed in the S. crispus sub-fraction-treated A431 after PDT. Western blot analysis indicated the expression of Bax and caspase-3 proteins was upregulated, while Bcl-2 was downregulated upon PDT.


The results in the present study suggest that S. crispus could be served as a potential source of photosensitizing agent for treatment of cancer via PDT.

Keywords: Strobilanthes crispus, photocytotoxic, PDT, pheophorbide-a, human skin squamous carcinoma (A431)

Oral 1.10

Clausenidin suppresses the production of VEGF and inhibits angiogenesis of HepG2 cells

Peter M Waziri¹*, Rasedee Abdullah², Nur Kartinee Kassim² and Jaafar Sani Muh’d¹

¹Kaduna State University, Kaduna Nigeria
²University Putra Malaysia, Selangor, Malaysia

Corresponding author: Peter M Waziri, petermwaziri@gmail.com


Metastasis of tumor cells is responsible for over 90% of cancer deaths globally. The production of vascular endothelial growth factor (VEGF) promotes metastasis and immortality of cancer cells. Up till date, there is still no clinical drug that effectively prevents the production of VEGF in tumor cells. Clausena excavata Burm f (local name: cherek hitam) is a shrub that is used locally for the treatment of cancer tumors in Malaysia and other countries of South East Asia. The current study evaluated the effect of clausenidin isolated from the chloroform extract of Clausena excavata Burm. f on the production of VEGF in liver cancer (HepG2) cells.


The anti-angiogenic effect (production of VEGF) in the clausenidin-treated cells was monitored using Western blot assay and the anti-apoptotic effect of the pure compound was monitored using transmission electron microscopy (TEM). Prior to the Western blot analysis, cells were treated with 5, 15, 30 and 40 µg/mL of clausenidin and in the TEM analysis, cells were treated with the IC50 (7.7 µg/mL) of clausenidin at 24, 48 and 72 h.


The Western blot analysis showed that clausenidin treatment significantly reduced (p<0.05) the production of VEGF that causes metastasis of liver cancer cells in a dose-dependent manner. In addition, we observed apoptosis to be one of the routes of cell death in the clausenidin-treated cells. The TEM analysis revealed margination and condensation of chromatin, nuclear fragmentation, formation of lipid droplets and convolution of nuclear outline (at all treatment points) which confirmed apoptosis of the clausenidin-treated HepG2 cells.


Clausenidin can be used as an anti-angiogenic and anti-apoptotic agent for the treatment of liver cancer.

Keywords: Clausenidin, VEGF, apoptosis, cancer.

Oral 1.11

Morphological Analysis of MCF-7 cells Treated with Chalcone Derivatives

Fatimah Hashim¹,²*, Wan Mohd Khairul Wan Mohamed Zin¹, Mas Mohamad¹, Nurul Shazwani Mohd Norhadi Shah¹, Syed Ahmad Tajudin Tuan Joharil³, Adibah Izzati Daud¹,, and Rafizah Rahamatullah¹,

¹School of Fundamental Science, Universiti Malaysia Terengganu, 21030 Kuala Nerus, Terengganu, Malaysia
²Institute of Marine Biotechnology, Universiti Malaysia Terengganu, 21030 Kuala, Terengganu, Malaysia
³Faculty of Bioresources and Food Industry, Universiti Sultan Zainal Abidin, 22200 Besut, Terengganu, Darul Iman, Malaysia
⁴Faculty of Engineering Technology, Universiti Malaysia Perlis, Level 1, Block S2, UniCITI Alam Kampus, Sungai Chuchuh, 02100 Padang Besar, Perlis, Malaysia

Corresponding author: Dr. Fatimah Hashim, fatimah.h@umt.edu.my


Three chalcone derivatives substituted with nitro, trifluomethyl and cyano chalcone have been synthesised and characterised. They were then evaluated for their cytotoxicity potential in inhibiting the growth of MCF-7 cells. This is based on the presence of conjugated double bond within the molecular framework by having delocalised electrons system on both aromatic rings.


Fifty percent inhibition concentration (IC50) of all chalcone derivatives were obtained via the MTT assay. The morphological changes of the MCF-7 were then observed under light microscopy.  Apoptosis in cells were examined through fluorescence acridine orange and propidium iodide staining. Phosphatidylserine (PS) exposure were determined by Annexin V staining and observed under fluorescence microscopy.


Nitro and trifluoromethyl chalcone compounds showed active activity against MCF7 with IC50 values 14.75 and 13.75 µg/ml, respectively. Cyano chalcone was, however, found to be inactive (IC50 > highest tested concentration). Both active compounds have triggered morphological changes to the MCF-7 cells which included cell shrinkage, rounded and clumping. The compounds also induced apoptosis with cell blebbing. Other apoptosis indicator included PS localisation on the external phospholipid bilayer.


Based on cell morphological changes observed in the present study, nitro and trifluoromethyl chalcone compounds could potentially cause apoptosis in MCF-7 cells.

Keywords: Chalcone, conjugated rings, MCF-7 cells, apoptosis, Phosphatidylserine

Oral 1.12

Cell Cycle Arrest by Cowanin on T47d Breast Cancer Cell Line

Fatma S. Wahyuni1*, Dessy Arisanty1 and Lusiana N. Yusra1

1Andalas University, Indonesia

Corresponding author: Prof. Fatma S. Wahyuni, fatmasriwahyuni@gmail.com


Breast cancer is one of the most common cause of cancer deaths each year. The incidence of cancer is very closely related to cell cycle. Cancer is a disease in which there are irregularities in the regulation of  cell cycle. Treatment of cancer depends mainly on chemotherapy. Nevertheless, anticancer agents lack selectivity and are often compromised by onset of resistance. The search for new drugs from natural sources is required. This study investigated the effect of cowanin from the stem bark of asam kandis (Garcinia cowa Roxb.) against T47D breast cancer cell cycle by flow cytometry method. Previous study showed that cowanin induced cytotoxic effect at IC50 11.68 µg/mL against T47D breast cancer cells. This study involved three groups, namely the control group, treatment group 1 (cowanin at 1x IC50), and treatment group 2 (2x IC50). Flow cytometry analysis showed that cowanin inhibited T47D breast cancer at G0-G1 phase. Concentration of 1x IC50 yielded an average percentage of inhibition 54.12% whereas the concentration of 2x IC50 produced an average percentage of inhibition 67.72%. Statistical analysis with one way ANOVA showed that there is significant difference between each treatment group in the percentage of inhibition on G0-G1 phase. The results indicated that cowanin induced cell cycle arrest in T47D cells, suggesting its potential as a new chemotherapeutic agent

Keywords : cowanin, cell cycle, Garcinia cowa, T47D, flow cytometry


Oral 2.1

CRISPR/Cas9 Mediated BFL-1 Knock-out in Nasopharyngeal Carcinoma (NPC) Cell Lines

Siti Fairus Abdul Rahman1*, Nethia Mohana-Kumaran1, Mohd Ghows Mohd Azzam1, Nur Adelina Mohd Norudin2 and Kalaivani Muniandy1

1University of Science, Malaysia, Malaysia
2Malaysian Institute of Pharmaceuticals and Nutraceuticals, NIBM, Malaysia

Corresponding author: Nethia Mohana-Kumaran, nethiakumaran@usm.my


BCL-2 family proteins which are divided into pro- and anti-apoptotic proteins are critical regulators of the intrinsic apoptosis pathway. The anti-apoptotic proteins are upregulated in many cancers and have become attractive therapeutic targets. ABT-263, which inhibit anti-apoptotic proteins BCL-2, BCL-XL and BCL-w, has little effect on Nasopharyngeal carcinoma (NPC) cells, made it clear that other anti-apoptotic proteins such as MCL-1 or BFL-1/A1 are important in survival of NPC cells. Previous studies show that Epstein-Barr virus proteins LMP1 and EBV nuclear antigen 2 (EBNA2) increase the level of BFL-1 mRNA in Burkitts lymphoma cell lines, promoting its survival. Given that NPC is closely associated with EBV infection and most NPC patients in Malaysia are positive for EBV infection, we believe that BFL-1 together with other anti-apoptotic proteins could contribute to NPC survival in a collaborative manner. In this study, the functional role of BFL-1 is determined by knocking-out the gene using the CRISPR/Cas9 technology. The long-term goal of the study is to test the sensitivity of the BFL-1 knockout NPC cells to specific BCL-2 inhibitors to dissect the contribution of each of the anti-apoptotic proteins for NPC survival.


The BFL-1 gene was knocked-out in two NPC cell lines, HK1 and C666-1 cell line using the CRISPR/Cas9 genome editing tool.


Two BFL-1 guide RNAs were designed and cloned into vector PX458 (pSpCas9(BB)-2A-GFP) and successful ligation was confirmed with PCR and Sanger sequencing. BFL-1 recombinant plasmids were transfected into NPC cell lines HK-1 and C666-1 using the TransIT-X2 Dynamic Delivery System from MirusTM. Single cell clones of the BFL-1 knockout NPC cells were generated through serial dilution.


To this end, targeting BFL-1 by CRISPR/Cas9 did not cause rapid cell death indicating that NPC cells do not solely depend on BFL-1 for survival and require inhibition of other anti-apoptotic proteins to mediate cell death.

Keywords: Nasopharyngeal carcinoma, CRISPR/Cas9, BFL-1, anti-apoptotic proteins and apoptosis

Oral 2.2

Functional characterization of FBXW7 mutations in colorectal cancer 

Najwa F. Md Yusof1, Nurul-Syakima Ab Mutalib1*, Luqman Mazlan2 and Rahman Jamal1

1UKM Medical Molecular Biology Institute (UMBI), Malaysia
2Department of Surgery, Faculty of medicine, University of Malaya, Malaysia

Corresponding author: Nurul-Syakima Ab Mutalib, syakima@ppukm.ukm.edu.my


The somatic mutational landscape of colorectal cancer (CRC) has been widely profiled, yet is inadequately characterized functionally. One of the most frequent somatic mutations in CRC is in the F-box WD repeat domain-containing-7 (FBXW7) gene. FBXW7 encodes for F-box protein, the substrate-recognition subunit of the SKP1–Cullin1–F-box (SCF) E3 ubiquitin ligase. Mutations in the FBXW7 could inhibit the ubiquitin-mediated degradation of various oncoproteins, leading to the accumulation of its oncogenic substrates and, subsequently, cancer progression. Those mutations are also associated with chemotherapy resistance, although the mechanisms are poorly understood.


FBXW7 mutations were detected via whole genome sequencing (WGS) and validated using Sanger sequencing. FBXW7 expression in matched tumour-normal samples was analysed using real-time quantitative PCR (RT-qPCR). DDK-tagged expression plasmids of FBXW7 wild-type (wt), R479Q and G654fs mutations were introduced into 293T, HCT116, and SW1463 cell lines, and the transfected cells were assayed for FBXW7 gene expression, cell viability, response to 5-fluorouracil (5-FU) treatment, and protein expression using targeted antibody arrays.


FBXW7 expression is down-regulated in CRC tissues compared to normal tissues. Introduction of R479Q and G654fs mutations into all the cell lines resulted in higher cell proliferation than in wt-FBXW7-transfected cells. On the other hand, restoring wt-FBXW7 in SW1463 cell line carrying the R479Q mutation inhibited cell proliferation and increased sensitivity to 5-FU treatment. Furthermore, ubiquitination levels of 38 proteins were decreased in both R479Q and G654fs transfected samples compared to the wild-type. The mutations were also observed to cause dysregulation of cancer-related proteins that are related to ubiquitin-mediated degradation.


Our results demonstrate that restoration of wt-FBXW7 in SW1463 influences cell viability and sensitizes SW1463 to 5-FU. The results also suggest FBXW7 inactivation caused by R479Q and G654fs mutations may contribute to the decreased levels of protein ubiquitination and dysregulation of cancer-related proteins, which could be the putative targets of FBXW7.

Keywords: FBXW7, colorectal cancer, 5-fluorouracil, gene expression, protein expression

Oral 2.3

MG63 human osteoblast tumour cells exhibit different behaviour when treated with encapsulated simvastatin PLGA microspheres: A morphological studies 

Nur Mohamed1, Nur A. Mohamed1, 2*, ANDREW MORRIS2, NASHIRU BILLA2 and Kevin Shakesheff2

1Universiti Teknologi MARA, Malaysia
2University of Nottingham, United Kingdom

Corresponding author: Miss. Nur A. Mohamed, , Universiti Teknologi MARA, Shah Alam, Malaysia, alianahidayah@salam.uitm.edu.my


Simvastatin (SIM), a widely used cholesterol-lowering drug, also exhibits tumor-suppressive potentials in several types of malignancy. The statin family of drugs preferentially triggers tumor cell apoptosis by depleting mevalonate pathway metabolites farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP), which are used for protein prenylation, including the oncoproteins of the RAS superfamily. Direct treatment of simvastatin potentially showed toxicity in bone tumor. By encapsulating SIM in microspheres, the drug delivery with sustained release and less toxicity can be achieved.


The preparation of SIM-loaded microparticles was based on solvent evaporation technique. Subsequently, the MG63 human osteosarcoma cells were cultured in the presence of SIM-loaded microparticles for 24, 48 and 72 hours, and their proliferation was assessed by MTT assay and further analysed with flow cytometry. Cells grown in blank microparticles served as the control. Upon the treatment, cells from the culture were prepared for light, fluorescent and scanning electron microscopy studies.


Interestingly, cultured MG63 cells showed apoptotic effects from 0.097mg/ml to 0.4 mg/ml however starting to increase proliferation from 0.4 mg/ml to 12.0 mg/ml. Results showed spheroid morphology with numerous secretion vesicles accumulated on the surface, observing no cytoplasmic projections with intercellular connections. Cells cultured with simvastatin encapsulated microparticles between the concentration of apoptotic effects had a polygonal and spindle-shaped morphology, with cytoplasmic projections that interconnected cells.


The apoptotic effect of SIM-loaded microparticles was significant at the early treatment and increased as the concentration of the microparticles increased. The morphologically destructed MG 63 was reverted by SIM-loaded microparticles. This may suggest the potential of SIM-loaded microparticles in cell stress and stabilizing the osteogenic renewal processes. However, as the concentration increased, the cellular activities was less sensitive and contributed to the proliferation of MG 63 cells. Interestingly, these proliferated cells produce an accumulation of cytosolic lipid droplets which indicate antiproliferative effects in malignant cells.

Keywords: Simvastatin-loaded microparticles, MG63, proliferation, human osteosarcoma, bone metabolism and cancer

Oral 2.4

Controversial Truth: Human Pancreatic Cancer Cell Line Homes Cancer Stem Cells

Yuan Han Teh1, Rajesh Ramasamy2 and Johnson Stanslas1*

¹Pharmacotherapeutics Unit, Department of Medicine, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia
²Immunology Unit, Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia

Corresponding author: Johnson Stanslas, jstanslas@yahoo.co.uk


Pancreatic cancer stem cells (pCSCs) are the culprit of pancreatic cancer recurrence. Disputable presence of pCSCs in continuous cell line has led to an extensive application of patient-derived pCSC culture for in vitro evaluation of anti-pCSC agents. Nevertheless, the procurement of patients’ tumour biopsies can be challenging to some researchers. Here, we investigated the presence of CSCs in pancreatic cancer cell lines (PANC-1, MIA PaCa-2, Capan-2, and BxPC-3).  


Dose-response effect of gemcitabine and vismodegib on pancreatic cancer cell lines was evaluated by MTT cell viability assay. Immunophenotyping was achieved via flow cytometry. Limiting dilution spheroid formation assay and Extreme Limiting Dilution Analysis (ELDA) software determined in vitro tumourigenicity of Capan-2 cells. Tumourspheres were treated with gemcitabine and vismodegib, followed by counting intact tumourspheres.


Capan-2 was the most resistant cell line to gemcitabine (IC50 > 100 µM). This profound chemoresistance coincided with the presence of a cell subpopulation (3.8%) that co-expressed pCSC surface markers (CD44, CD24, and CD133), which was undetectable in other cell lines. ELDA software estimated that 1.45 − 4.15% of Capan-2 cells were tumourigenic. These findings collectively imply that CD44+CD24+CD133+ subpopulation is potentially pCSCs, which survive chemotherapy and initiate recurrence. Capan-2 tumourspheres demonstrated molecular features of pCSCs as reported previously. GLI1 expression was down-regulated in these tumourspheres, suggesting an independence of canonical Hedgehog signaling. This finding explains an unexpectedly moderate response of CD44+CD24+CD133+ cells (p = 0.738) and tumourspheres (IC50 > 20 µM) to vismodegib treatment that targets canonical Hedgehog signaling. The integrity of tumourspheres was remarkably irresponsive to gemcitabine (IC50 > 100 µM), which is in line with an enrichment of CD44+CD24+CD133+ subpopulation (p = 0.002).


Capan-2 human pancreatic cancer cell line harbours CD44+CD24+CD133+ subpopulation that behaves like pCSCs, thus is an attractive substitute for patients’ tumour biopsies for anti-pCSC drug discovery.

Keywords: Pancreatic cancer stem cells, Pancreatic cancer cell lines, Chemoresistance, Tumourigenicity and Hedgehog signaling

Oral 2.5

Transcriptional Gene Expression Profiles on Nasopharyngeal Carcinoma/HK1 Cell Line with siRac1-Mediated Knockdown

Rabiatul Basria S.M.N. Mydin¹*, Simon I. Okekpa¹,², Emmanuel Jairaj Moses³, Adam Azlan³, Gurjeet Kaur4 and Yusri Musa¹

¹Oncological and Radiological Sciences Cluster, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Kepala Batas, Pulau Pinang, Malaysia.
²Department of Medical Laboratory Science, Faculty of Health Sciences, Ebonyi State University, Abakaliki, Ebonyi State, Nigeria.
³Regenerative Medicine Cluster, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Kepala Batas, Pulau Pinang, Malaysia.
4Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Pulau Pinang, Malaysia.

Corresponding author: Dr. Rabiatul Basria S. M. N. Mydin, rabiatulbasria@usm.my


In Asia, nasopharyngeal carcinoma (NPC) is one of the leading head and neck cancer and usually diagnosed at late stage with poor prognosis. The critical regulatory pathways in NPC malignancy, resistance behavior and recurrence cases are still not clear. Rac1 has become a promising therapeutic target for cancer associated with reactive oxygen species-mediated cell killing.


This work profiled a set gene expression associated in NPC malignancy phenotype via siRac1 (Small interfering RNA for Rac1 gene) activities. HK1 cell line was given by Professor George Tsao from University of Hong Kong represents the differentiated squamous nasopharynx carcinoma.


Findings from RT-qPCR results revealed gene regulations kindred in cell cycle and apoptotic regulators such as CDC42, BCL 2, TGF-β1, CDH1, BAX, MAPK14 and PIK3CA. Furthermore, significant regulation of increased AKT1 expression at 2.15-fold (p-value ˂0.01) and decreased KRAS expression at 0.66-fold (p-value ˂0.043) were observed and might involve in suppressing the malignancy phenotype.


This study suggests that NPC/HK1 cells with siRac1-mediated knockdown may be involved in anti-metastatic role and pro-apoptotic pathways. Further understanding on these mechanisms are crucial in developing an efficient NPC therapeutic target.

Keywords: Nasopharyngeal carcinoma, siRac1, RNAi therapeutics, Reactive oxygen Species-Mediated Cell Killing, Malignancy Phenotype

Oral 2.6

Elucidating the role of microRNA expression as a post-transcriptional mechanism in the carcinogenesis of air pollution-related lung cancer

Maizatul Syafinaz Shahadin¹, Nurul Syakima Ab Mutalib¹*, Mohd Talib Latif², Mohamed Faisal Abd Hamid³, Rahman Jamal¹ and Tidi Maharani Hassan4

¹UKM Medical Molecular Biology, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia
²Institute for Environmental and Development, Universiti Kebangsaan Malaysia, Selangor, Malaysia
³UKM Medical Centre, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia
4Beaumont Hospital and Our Lady of Lourdes Hospital, Dublin, Ireland

Corresponding author: Nurul Syakima Ab Mutalib, syakima@ppukm.ukm.edu.my


Previous epidemiological studies have reported positive associations between ambient air pollution and lung cancer, and henceforth, air pollution has been classified as group 1 carcinogen in humans. However, there are limited studies pertaining the molecular pathogenesis of air-pollution lung cancer tumourigenesis, including the role of noncoding RNA called microRNA (miRNA). Dysregulation of miRNAs expression is shown to be correlated with cancer, suggesting their potentials as the biomarker and therapeutic agents.


miRNA expression profiling was performed using real-time PCR in 10 non-smoker lung adenocarcinoma patients with low (n=5) and high (n=5) exposure to air pollution. The patients were stratified based on the residential address, level of particulate matter and anthropometric activities such as occupation, transportation and daily ambient exposure. We further validated the selected miRNAs in 22 lung adenocarcinomas (low, n=10; high, n=12), followed by in silico analyses to identify the putative targets and pathways regulated by the differentially expressed miRNAs.


The profiling analyses revealed 27 upregulated miRNAs in the high exposure group. miR-15b-5p (p=0.0269), let-7a-5p (p=0.0426), miR-151a-5p (p=0.0449), miR-222-3p (p=0.0446), and let-7f-5p (p=0.0333) were the most significantly upregulated miRNAs. miR-15b-5p (p=0.0244) and miR-222-3p (0.0378) remained significant in the validation set. Pathway enrichment analyses based on target prediction by TargetScan and microT-CDS showed that all of the significantly upregulated miRNAs were associated with MAPK signalling and Wnt signalling pathway. Several miR-15b-5p targets in MAPK signalling pathway have not been validated yet, suggesting a novel finding in air pollution-related lung cancer.


The upregulation of miR-15b-5p and miR-222-3p may be crucial in the tumourigenesis of air pollution-related lung cancer via the dysregulation of multiple cancer related pathways.

Keywords: Air pollution, non-smoker lung cancer, molecular mechanism, biomarkers, microRNA

Oral 2.7

Identification of Fusion Genes in Pediatric Relapsed AML: A Preliminary Finding

Nadiah Abu¹, Siti Hawa Osman¹, Habsah Aziz¹, Hamidah Alias², Rahman Jamal¹*

¹UKM Medical Molecular Biology Institute (UMBI), UKM Medical Center, Universiti Kebangsaan Malaysia
²Department of Pediatrics, UKM Medical Center, Universiti Kebangsaan Malaysia

Corresponding author: Rahman Jamal, rahmanj@ppukm.ukm.edu.my


Pediatric Acute Myeloid Leukemia (AML) is a highly complex and heterogenous disease. To date, the mechanisms of its regulatory network, particularly during relapse, have not been elucidated. One of the causes of its pathogenesis is through chromosomal rearrangements, which may result in fusion genes and transcripts. The pattern of expression of fusion genes in relapsed AML is unclear, especially as predictive biomarkers for response or relapse. The best approach is to profile the patients’ cells at diagnosis, remission and relapse.  


Total RNA was extracted from the bone-marrow derived mononuclear cells of three pediatric AML patients at different stages; diagnosis, remission and relapse. One patient (PAML1) had a normal karyotype, while the other two patients (PAML2 and PAML3) were positive for the t(8 t(8;21) (q;22 q;22) translocation. We performed deep, paired-end RNA sequencing on these AML trios. Putative fusion transcripts were identified and prioritized using the SOAPFuse algorithm. Further visualization of the fusion genes was conducted using Circosplot.


PAML1 had several novel fusion transcripts, among which, the fusion transcript NTUM2A-AS1>>MINPP1 was persistent at all three stages of diagnosis, remission and relapse. Commonly reported fusion genes involving the MLL gene (MLL>>MLLT4) were also identified in the diagnosis and the relapsed stages. In PAML2, there were no persistent fusion transcripts identified in all three stages. However, as predicted, the RUNX1>>RUNXT1 fusion transcripts were detected in the diagnosis stage and later, reappeared in the relapsed stage. This was also observed for the RRN3P3>>DARS fusion gene. In PAML3, the fusion gene PARP6-JHD1MD persisted in all three stages. Similarly, the RUNX1>>RUNXT1 fusion genes were also detected in PAML3. Interestingly, the fusion gene C15orf57>>CBX3 was found to be expressed in the relapsed stage of all three samples. However, no common fusion genes were detected in the diagnosis and remission stage of all three samples.


Based on our preliminary findings, fusion genes are selectively expressed at different stages of the disease and in different patients. However, further validation as well as understanding the true biological function of the fusion genes should be investigated. This should be further correlated with the respective gene expression profile, as well as its mutational status.

Keywords: Pediatric AML, Relapse, Fusion Genes, Transcriptome, RNA-Seq

Oral 2.8

Combined use of epithelial membrane antigen and nuclear matrix protein 52 as sensitive biomarkers for detection of bladder cancer

Attallah AM¹, El-Far M², Abdallah SO³, El-Waseef AM², Omran MM4, Abdelrazek MA¹, Attallah AA¹, Saadh MJ¹, Radwan M¹, El-waffaey KA¹, and Abol-Enei H⁵.

¹Research and Development Department, Biotechnology Research Center, New Damietta City – Egypt.
²Chemistry Department, Faculty of Science, Mansoura University, Mansoura – Egypt.
³Chemistry Department, Faculty of Science, Cairo University, Giza – Egypt.
4Chemistry Department, Faculty of Science, Helwan University, Cairo – Egypt.
Department of Urology, Urology-Nephrology Center, Mansoura University, Mansoura – Egypt.

Corresponding author: Dr. Mohammed Saadeh, msaadeh@philadelphia.edu.jo; mjsaadh@yahoo.com


The advent of noninvasive urine-based markers as well as other novel modalities has yielded improved diagnostic accuracy. However, the new markers failed to reach higher sensitivity and specificity. We therefore evaluated the potential role of epithelial membrane antigen (EMA) and nuclear matrix protein 52 (NMP-52) singly and combined as noninvasive biomarkers for the detection of bladder cancer (BC).


A total of 160 individuals including 66 patients with BC, 54 patients with benign urologic disorders and 40 healthy volunteers were investigated. Urinary EMA at 130 kDa and NMP at 52 kDa were identified, purified and quantified by Western blot, electroelution and enzyme-linked immunosorbent assay (ELISA). The diagnostic performance of each biomarker and their combination were compared using area under receiver operating characteristic curves (AUC).


Mean urinary EMA, 2.42 µg/mL, and NMP-52, 17.85 µg/mL, were significantly elevated in patients with BC compared to controls, 1.18 and 3.44 µg/mL, respectively (p < 0.0001). The combined use of these markers yielded values which were increased 4.4- and 13.7-fold in the benign and malignant disease groups, respectively, with respect to the normal group. The values of EMA and NMP-52 were significantly higher in patients with higher-grade tumors than those with lower-grade tumors (p < 0.0001). Moreover, this combination could predict all BC stages and grades with 0.91 AUC, 94% sensitivity and 80% specificity.


EMA and NMP-52 in combination could be promising noninvasive biomarkers for BC detection.

Keywords: Bladder cancer, Biomarker, nuclear matrix protein 52, epithelial membrane antigen.

Oral 2.9

Epigenome analysis of colorectal cancer: A genome wide approach

Muhiddin Ishak¹, Nurul Syakima Ab Mutalib¹*, Najwa Farhah Mohd Yusof¹, Sazuita Saidin¹, Isa Mohamed Rose², Luqman Mazlan³, Ismail Sagap³, Rahman Jamal¹

¹UKM Medical Molecular Biology Institute (UMBI), National University of Malaysia, Kuala Lumpur, Malaysia
²Department of Pathology, Faculty of Medicine, National University of Malaysia, Kuala Lumpur, Malaysia
³Department of Surgery, Faculty of Medicine, National University of Malaysia, Kuala Lumpur, Malaysia

Corresponding author: Nurul Syakima Ab Mutalib, syakima@ppukm.ukm.edu.my


Colorectal cancer (CRC) contributes to around 1.36 million of the total cases worldwide. In Malaysia, the incidence rate of CRC is 21.3 cases per 100,000 population. Many molecular pathways are involved in carcinogenesis which includes DNA methylation. Genes with high levels of 5-methylcytosine (hypermethylation) in the promoter regions are associated with gene silencing and may alter the signalling pathway that contribute to CRC tumourigenesis. Subsequently, genes with low levels of 5-methylcytosine (hypomethylation) have also been implicated in CRC progression through tumour-suppressor genes or act as oncogenes. Many studies have also revealed the involvement of enhancer regions in the regulation of gene expression, hence the demand for a more comprehensive tool for methylation profiling.


Genomic DNA was extracted from 12 paired matched samples collected from UKM Medical Centre (UKMMC). Bisulfite conversion was performed and the bisulfite converted DNA was subjected to microarray using Human Infinium Epic Beadchip Array. The data was analyzed using Bioconductor-ChAMP V2.8.1 and Genome Studio V1.8.


There were 25170 significantly differentially methylated probes (p≤ 0.05), with 7254 being hypermethylated and 17916 being hypomethylated. ADHFE1, C1orf70 and CMTM3 were the top three hypermethylated genes whereas AMPH, CHST10 and MYBPC3 were the top three hypomethylated genes. Pathway enrichment analysis revealed that majority of the hypermethylated genes were involved in neuroactive ligand receptor, cancer pathways, calcium pathway, and cAMP signalling, amongst others. Meanwhile, the hypomethylated genes were involved in olfactory transduction, neuroactive ligand receptor, PI3K-Akt signalling and focal adhesion. Out of this differently methylated genes, 715 genes were found in Phantom4 enhancers, 355 genes in Phantom5 enhancers and 2214 genes at open chromatin.


This is the first study that investigated the methylation profile of local CRC patients using the latest platform assay. The new knowledge from this study can be utilized to increase our understanding of CRC methylomics further.

Keywords: Colorectal cancer; epigenetics; DNA methylation; enhancer; open chromatin; MethylationEPIC

Oral 2.10

Transcriptome Profiling of Pediatric Relapse Acute Myeloid Leukemia (AML)

Hawa Osman¹, Nadiah Abu¹, Hamidah Alias¹, Wan Fahmi Wan Mohamad Nazarie¹ and Rahman Jamal¹*

¹UKM Medical Molecular Biology Institute (UMBI), Kuala Lumpur, Malaysia
²Department of Paediatric, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia

Corresponding author: Rahman Jamal, rahmanj@ppukm.ukm.edu.my


Acute myeloid leukemia (AML) is the second most common leukemia type among children, and nearly 50% of pediatric AML patients will relapse and succumb to the disease. Clinically, relapsed AML patients respond poorly to the standard chemotherapy regimens, therefore there is a need to investigate what are the genes that could define relapsed AML in order to predict and improve patients’ survival via targeted therapy.


Transcriptome sequencing (RNA-seq) was performed on trio diagnosis, remission and relapse samples of three pediatric AML patients. The differential gene expressions in all three samples were analyzed via the edgeR method. The gene ontology and pathway analysis were performed using the KEGG and GO database. Furthermore, the expression data for several selected genes were validated using real-time quantitative PCR in 26 diagnosis samples and 7 relapse samples. The expression ratios were calculated using the ΔΔCT method after prior validation of the method for each target. The functional activity of the selected gene was further validated in an in vitro setting.


The regulation of the differentially expressed genes varied between patients. A set of differentially expressed genes for the relapse vs diagnosis comparison was then obtained. These genes were enriched in the cell cycle-related pathway. Several up-regulated genes were selected for validation. Validation results of two selected genes (ADGRL3 and SELIL2) showed that both were up-regulated in relapse samples as compared with diagnosis samples, which was in concordance with the RNA-seq results.


Since little is known about the molecular mechanism(s) underpinning relapsed AML, this study has allowed better understanding on the gene expression pattern in relapse pediatric AML. We are also able to further comprehend the molecular pathways contributing towards relapse pediatric AML.

Keywords: pediatric AML, transcriptome, RNA-seq, relapse, differentially expressed genes

Oral 2.11

Annona muricata (Soursop) Leaves Extract Increased Caspase 3 Expression in Patient-derived WHO III Nasopharyngeal Cancer Cell

Ersty Istyawati¹, Pudji Rahaju¹, and Ahmad Dian Wahyudiono¹*

¹Department of Otorhinolaryngology Head and Neck Surgery, Medical Faculty of Brawijaya University – Saiful Anwar General Hospital, Malang Indonesia.

Corresponding author: Ahmad Dian Wahyudiono, dian_w.fk@ub.ac.id


Nasopharyngeal cancer is the most common head and neck cancer in Indonesia. Annona muricata leaves extract contains annonaceous acetogenin, which have emerged as potentially promising anti cancer drugs. The purpose of this study is to investigate the effect of soursoup (Annona muricata) leaves fraction  in inducing caspase 3 expression in  cell cultured from biopsy tissue of nasopharyngeal cancer WHO III.


This study is a true experimental study that use post test only control group design. This study uses cell culture and divided into 5 group treatments. Soursop leaves extraction uses ethanol 95%. Bioactive agents used are fraction materials from crude extract using hexane and methanol. There are 5 group dosage 0, 31.25 µg/ml, 62.5 µg/ml, 125 µg/ml dan 250 µg/ml . The caspase 3 expression was detected by caspase 3 Elisa assay kit observed in 6, 12 and 24 hours.


Expression of caspase 3 in treatment group is higher than control group. The expression increased by time and the highest is in 24 hours after treatment. Increased in dosage was also followed by increased in caspase 3 expression especially at doses of 125 µg/ml and 250 µg/ml.


Annona muricata leaves extract induced caspase 3 expression in  WHO III nasopharyngeal cancer cultured cells

Keywords: Annona muricata, acetogenin, WHO III nasopharyngeal cancer cells.

Oral 2.12

The Development of a Synthetic scFv Monoclonal Antibody Targeting Pro-oncogenic AGR2

Aiman Mohtar¹,²*, Low Teck Yew¹, Rahman Jamal¹, Borek Vojtesek³, and Ted R. Hupp ²,³,4

¹National University of Malaysia, UKM Medical Molecular Biology Institute (UMBI), Kuala Lumpur, Malaysia
²University of Edinburgh, Institute of Genetics and Molecular Medicine, Edinburgh, Scotland, United Kingdom
³Regional Centre for Applied Molecular Oncology, Masaryk Memorial Cancer Institute, Brno, Czech Republic
4University of Gdansk, International Centre for Cancer Vaccine Science, Gdansk, Poland

Corresponding author:M. Aiman Mohtar, m.aimanmohtar@ppukm.ukm.edu.my


AGR2 is an endoplasmic reticulum (ER)-resident protein that belongs to the protein disulphide isomerase (PDI) family. These enzymes facilitate the rearrangement of disulphide bonds during protein maturation in the ER. AGR2 has emerged as a clinically relevant anti-tumour target since it possesses pro-oncogenic features, promotes metastasis and is overexpressed in diverse types of cancer. One unique molecular function of AGR2 is to drive certain oncogenic client proteins maturation in the ER, such as receptor EpCAM, via a specialised docking site.  

Methods and Results 

We developed synthetic single-chain variable fragment (scFv) monoclonal antibodies targeting AGR2. First, recombinant AGR2 protein was used as an antigen for screening a naïve canine phage-scFv library for specific scFvs. After four rounds of screening, nine AGR2-binding scFvs were identified; while subsequent epitope mapping highlighted the presence of four different classes of scFvs. We next selected one scFv that binds strongly to the N-terminal of AGR2 (scFv4) for further studies. We re-designed scFv4 such that the new scFv4 construct contains a C-terminal CD20 epitope tag, and ER postcodes for entry into ER as tools to manipulate the proteins inside the ER.  Although all synthetic scFv scaffolds were expressed in AGR2-negative cells, the AGR2-binding scFv with an ER-postcode was selectively degraded in AGR2-positive cells. These data suggest that the expression of scFv scaffolds can be impacted in an AGR2-dependent manner and the presence of AGR2 in cells can suppress the production of a high-affinity binding protein when it is destined for the ER.


This study demonstrates the successful construction of a novel AGR2-binding scFv scaffold that can be expressed in mammalian cells. These tools were used to show the key determinants that regulate the ability of AGR2 to mediate client protein maturation in the ER and can be potentially manipulated for the development of human monoclonal antibodies for therapeutic treatment of cancer.

Keywords: anti-cancer, endoplasmic reticulum, chaperone, secretory pathway, protein disulphide isomerase


Oral 3.1

The Stepping Force Method: Assessment of Pain in Rat Models

Mun Fei Yam¹,²*, Siew Li Khoo², Yean Chun Loh², Rusliza Basir¹

¹Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia
²School of Pharmaceutical Sciences, Universiti Sains Malaysia, 11800 Minden, Pulau Pinang, Malaysia

Corresponding author: Mun Fei Yam, yammunfei@yahoo.com


Pain research has been widely performed in recent years to measure the pain intensity for drug development. Among the in vivo pain model, pain intensity study based on normal behaviours of the animal imposed the least biased, most reliable and  more ethical approach in conducting pain research.


A new stepping force analgesic meter equipped with data acquisition and automatic stepping force identification function was fabricated. Intra-day and inter-day precision and accuracy of the analgesic meter were evaluated. In in vivo studies, carrageenan and CFA were used to induce ipsilateral hyperalgesia and arthritic

pain in rats, respectively. In hyperalgesia study, the stepping force was measured before and 4 days after induction (day 0) and 4, 7, 11 and 14 days after treatments onset. Hind paw thickness of the rats were also measured in conjunction to the stepping force measurement for validating the inflammatory state of the rats.


The validation studies for each detection sensors of stepping force analgesic meter also showed convincing results whereby intra-day and inter-day precisions were less than 2% and accuracy was 99.33–100.35. In hyperalgesia model, the ratio of stepping force of ipsilateral to contralateral of hind paw under treatment of indomethacin (2.5, 5 and 10 mg/kg p.o.) was significantly different at the 4 hours post induction as compared to the control rats. Meanwhile, in arthritic pain model, the ratio of stepping force between ipsilateral and contralateral of hind paw was significantly different at day 4 and 7 after treatment of prednisolone (2.5 mg/kg p.o.) as compared to the control rats. The ipsilateral paw thickness in CFA-induced arthritic model was correlated to the stepping force.


This study indicates that the stepping force analgesic meter is an effective tool for the assessment of pain within a more realistic setting, and may be useful for discerning therapeutic approaches considered worthy of further investigation using other arthritic animal models.

Keywords: stepping force, carrageen, arthritis, and CFA

Oral 3.2

Phytochemical Screening of White Oyster Mushroom (Pleurotus Ostreatus) Preparations

Santun Bhekti Rahimah¹,*, Dhiah Dianawaty Djunaedi², Arto Yuwono Soeroto³, Tatang Bisri⁴

¹Doctoral Programe, Padjadjaran University, Bandung, Indonesia
²Biochemistry Departemen, Padjadjaran University, Bandung, Indonesia
³Internal Medicine Departemen, Padjadjaran University, Bandung, Indonesia
⁴Anesthesia Departemen, Padjadjaran University, Bandung, Indonesia
Pharmacology Departemen, Bandung Islamic University, Bandung, Indonesia

Corresponding author: Santun Bhekti Rahimah, santunbr@gmail.com


Pleurotus ostreatus a popular commercial mushroom contains very high nutrients and bioactive compounds with therapeutic effects. The ethanolic extracts seem to be to the most active preparation. This study is aimed to compare the phytochemical analysis of fresh, dry and ethanolic extracts of  Pleurotus ostreatus..


The fresh (FPM), dry (DPM), ethanolic extract macerated with 70%  (EE70) and 96% ethanol (EE96) of Pleurotus ostreatus were used in the study. The phytochemical analysis was conducted using Dragendorf and Meyer reagents for alkaloids, FeCl3 for polyphenolic and tannins, Sawkosky method for steroids, Lieberman for triterpenoid, amyl alcohol reagent for flavonoids, foam test for saponin, NaOH reagent for Quinone and gelatin test for tannins and phenolic compounds.


The phytochemical screening showed that flavonoid, phenolic compounds, tannin, saponin, alkaloids, and steroids were detected in FPM, DPM, EE70 and also EE96. Alkaloid however, was not identified by Meyer Reagent in FPM and DPM. DPM and EE70 seemed to have the highest amount of saponin based on the foam formed. Meanwhile, steroids and flavonoids were detected at a higher level in EE96, based on strength of visible colour. However, triterpenoid and quinones could not be identified.


The process of drying Pleurotus ostreatus did not reduce the content of active substances. The slight differences in the phytochemical screening obtained in all samples may be due to the influence of water content and polarity of the active substances. The polar active substances seem to be more soluble in EE70 than EE96. The higher the bioactive substances in the preparation, the more significant the bio-therapeutic effects.

Keywords: dry plants materials, fresh plant materials, Pleurotus ostreatus, ethanolic extract phytochemical.

Oral 3.4

A Comparative Analysis of Mucosal Associated Invariant T Cells and Invariant Natural Killer T Cells

Zarina Zainudeen¹,²*, Aman Shah Majid³, Chris Willberg4

¹Nuffield Department of Clinical Neurosciences, John Radcliffe Hospital, University of Oxford, UK
²Advanced Medical and Dental Institute (AMDI), Universiti Sains Malaysia, Penang, Malaysia
³Centre for Natural Product and Angiogenesis Research/Department of Pharmacology, Faculty of Medicine, Quest International University, Ipoh, Malaysia
4Experimental Medicine Division, John Radcliffe Hospital, Headington, Oxford, UK

Corresponding author: Zarina Zainudeen, zarina.zainudeen@ndcn.ox.ac.uk


CD1d-restricted invariant natural killer T (iNKT) cells and MR-1-restricted mucosal associated invariant T (MAIT) cells are two subsets of non-conventional T cells, with conserved T-cell receptor (TCR) repertoire, functions, homing and cytokine production. Both cell types show remarkable differences in their abundance in humans and mice; iNKT cells are found at a higher frequency in mice than human, whereas the opposite situation is true for MAIT cells. This suggests that the T cell subsets can potentially replace the other. Since MAIT cells dominate CD161++CD8+T cell population, the aim of this study is to compare the common phenotypes and roles between CD161++CD8+ T and iNKT cells.


CD161++CD8+ T and iNKT cells were characterized in terms of chemokine and cytokine receptor expression and functionality. This study compared peripheral blood iNKT cells and MAIT cells from healthy individuals. Since CD161 is widely expressed by lymphocytes, flow cytometry was employed to analyse the two study populations at the single cell level, and distinguish them from other CD161 expressing cells.


Both cell types displayed preferential tissue-homing molecules, and different subsets of CD4/CD8 cell. Some of the Th17 characteristics were also found in iNKT cells, like IL23R and CCR6. In the present study, both the cells expressed granzyme A, K and perforin, with lower/undetectable expression of granzyme B. CD161++CD8+T cells were also observed to express higher levels of D prostanoid receptor (DP) 1 at the cell surface and at the mRNA level when compared to those seen in iNKT cells.


CD161++CD8+ T and iNKT cells are distinctive from conventional T cells.  Both these cells are selectively recruited to the site of inflammation and exhibit antimicrobial activity. Although some overlapping functions were observed between these populations, differences were detected in the cytokine expressions and cytolytic granule contents.

Keywords:  CD161++CD8+ T cells, iNKT cells, CCR6, IL23R,  DP1

Oral 3.5

Maslinic acid suppresses macrophage foam cell formation via inhibition of LDL oxidation and cholesterol efflux

Su Wen Phang ¹, Bee Kee Ooi ¹, Nafees Ahemad², and Wei Hsum Yap ¹*

¹School of Biosciences, Taylor’s University, Subang Jaya, Selangor, Malaysia
²School of Pharmacy, Monash University Malaysia, Petaling Jaya, Selangor, Malaysia

Corresponding author: Wei Hsum Yap, WeiHsum.Yap@taylors.edu.my


Accumulation of lipid-loaded macrophages, also known as foam cells, contributes to the development of atherosclerotic lesion. Development of novel therapeutic interventions targeting the process of macrophage foam cell formation may help to alleviate atherosclerosis. Maslinic acid (MA) is a novel natural pentacyclic triterpene that possess cardioprotective and anti-inflammatory properties. This study evaluates the potential of MA in targeting the process of foam cell formation, including monocyte recruitment, macrophage lipid uptake and accumulation, as well as cholesterol efflux.


An in vitro model of monocyte recruitment using THP-1 human monocytic cell line and human umbilical vascular endothelial (HUVEC) cells was developed for testing the effects of MA on TNF-α-induced monocyte adhesion and transendothelial migration. Meanwhile, foam cells were generated by incubating PMA-differentiated THP-1 macrophages with oxidized LDL (oxLDL) and MA was determined for its ability to reduce the uptake of ox-LDL, as quantified by Oil Red O staining and flow cytometric analysis. The study on cholesterol efflux on the other hand was investigated using fluorescently-tagged cholesterol (3-hexanoyl-NBD cholesterol) to determine the ability of MA in mediating cellular cholesterol efflux.


Our findings demonstrated that MA significantly suppressed monocyte adhesion to TNFα-induced HUVEC but did not affect transendothelial migration. We further showed that MA has the capability to inhibit macrophage foam cell formation which may result by inhibiting LDL oxidation, reducing uptake of ox-LDL and enhancing cholesterol efflux.


The findings of this study showed that MA targeted several points in the foam cell formation process, including its ability in reducing TNF-α-mediated monocyte adhesion to endothelial cells and LDL oxidation as well as enhancing cholesterol efflux.

Keywords: Foam cells, Maslinic acid, antioxidant, LDL oxidation, cholesterol efflux

Oral 3.6

Anti-arthritic Potential of Ardisia crispa Root (Myrsinaceae) In vitro and In vivo

Joan A. Blin1, Roslida Abd Hamid1*, Yoke Kqueen Cheah1 and Razana Mohd Ali2

1Department of Biomedical Sciences, Faculty of Medicine and Health sciences, University of Putra Malaysia, Malaysia
2Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia

Corresponding author: Dr. Roslida Abd Hamid, roslida@upm.edu.my


Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory joint disease characterised by excessive angiogenesis. Anti-angiogenesis could be a promising therapeutic strategy against RA. Ardisia crispa or “Mata Itik” is a local plant traditionally used for treating various illnesses. Its properties like anti-tumour, anti-inflammatory and anti-angiogenic have been reported in previous studies. However, its anti-angiogenic role against RA has yet to be investigated. Thus, the present study investigated the anti-angiogenic potential of hexane extract (ACRH), quinone rich fraction (QRF), and benzoquinone (BQ) isolated from the roots of the plant using cell-based assays in vitro. ACRH was also tested against in vivo model.


Cytotoxicity, tube formation, cell invasion, and cell apoptosis assays were performed using human umbilical vein endothelial cells (HUVEC) and human fibroblast-like synoviocytes for rheumatoid arthritis cells (HFLS-RA). In vivo study was performed using collagen-induced arthritis (CIA) in rats.


ACRH, QRF and BQ were highly cytotoxic against HUVEC (IC50 at 1.9 ± 0.2, 2.5 ± 0.8 and 1.7 ± 0.2 µg/mL, respectively). Nevertheless, ACRH and QRF were found to be less cytotoxic against HFLS-RA cell (4.0 ± 2.3 and 5.7 ± 0.9 µg/mL, respectively) after 24 hours of exposure. Interestingly, BQ remained highly cytotoxic against HFLS-RA cells with IC50 at 1.6 ± 0.5 µg/mL. All samples (0.05, 0.5 and 5 µg/mL) significantly (P<0.05) inhibited VEGF-induced HUVEC tube formation and IL-1β-induced HFLS-RA cell invasion in a concentration-dependent manner. Both ACRH and BQ exhibited late apoptotic activity in IL-1β-induced HFLS-RA cells at highest concentration (5 µg/mL). For CIA in rats, all doses of ACRH (10, 30, 100 mg/kg) showed insignificance reduction of ankle joint diameter, paw volume, arthritic scoring, and serum prostaglandin E 2 in treated group.


This study had uncovered the anti-angiogenic effect of Ardisia crispa root in suppressing angiogenesis against both HUVEC and HFLS-RA cells, even though its role in modulating angiogenic biomarkers in RA animal model has yet to be determined.

Keywords: Ardisia crispa root, Human Umbilical Vein Endothelial Cells (HUVEC), human fibroblast-like synoviocytes for rheumatoid arthritis cells (HFLS-RA), Rheumatoid arthritis, Angiogenesis, Collagen-induced arthritis rat

Oral 3.7

Probiotic supplementation during pregnancy and early childhood in the prevention of atopic dermatitis: A literature review

Kang Nien How¹*, Zee Wei Lai², Pui Ling Thong³

¹Department of Internal Medicine, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
²School of Biosciences, Faculty of Health and Medical Sciences, Taylor’s University Lakeside Campus, Subang Jaya, Selangor, Malaysia.
³Department of Paediatric, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.

Corresponding author: Kang Nien How, hkangnien@upm.edu.my


Atopic dermatitis (AD) is an increasingly prevalent chronic skin disorder that results in progression to atopic march. The hygiene theory has been postulated to have caused the rise in the incidence of atopic diseases among urban living. Early alteration of gut microbiota has been postulated to have immuno-regulatory effects, causing reduction in the incidence of atopic dermatitis. A literature search was therefore performed on randomized controlled trials that evaluate the effects of probiotic supplementation during pregnancy and early infancy in the prevention of atopic dermatitis.


To assess the effects of probiotic supplementation during pregnancy and early infancy in the prevention of AD in children, PubMed database was searched for randomized controlled trials that were published in English language.


Based on the search string, a total of 236 articles were identified. Two studies were further identified after listing the references of existing meta-analysis or systematic review. After going through the inclusion and exclusion criteria, a total of 14 studies comprising of 4712 subjects were included in the final analysis. Lactobacillus spp were used in 5 studies, Bifidobacterium spp. in 2 studies and a combination of probiotic strains in 8 studies. Probiotics were mainly initiated for 4-6 weeks before delivery and continued postnatally for an average of 6 months. Eight out of the 14 articles demonstrated a significant reduction in the incidence of atopic dermatitis in the probiotic arm. A combination of probiotic strains appeared to be superior when compared to a single strain probiotic in the prevention of atopic dermatitis.


Probiotic appeared to have a protective role in AD prevention if initiated during pregnancy and early infancy. Further analysis of the current data is however required.

Oral 3.8

Biotransformation of DYN 1-17 in inflamed nasal tissues: potential immunotherapeutics for chronic rhinosinusitis

Siti Sarah Fazalul Rahiman¹*, Michael Morgan², Paul Gray³, Paul Nicholas Shaw³,⁴ and Peter John Cabot⁴

¹School of Pharmaceutical Sciences, Universiti Sains Malaysia, Penang, Malaysia.
²School of Biomedical Sciences, The University of Melbourne, Victoria, Australia.
³Faculty of Medicine, The University of Queensland, Brisbane, Australia.
School of Pharmacy, The University of Queensland, Brisbane, Australia.

Corresponding author: Siti Sarah Fazalul Rahiman, sarahfazalul@usm.my


Dynorphin 1-17 (DYN 1-17) has a significant implication in inflammation. Upon release at inflammatory sites, this endogenous neuropeptide is subject to rapid metabolism, yielding an array of smaller fragments. DYN 1-17 and its fragments have been previously demonstrated to regulate diverse inflammatory responses during tissue injury. To date, biotransformation of DYN 1-17 in human disease condition has not yet been reported. This study aimed to explore the biotransformation of DYN 1-17 in nasal biopsies from chronic rhinosinusitis (CRS) patients and the fragmentation patterns were compared with those observed previously in inflamed rat paw tissues.  


DYN 1-17 (150 μM) was incubated with human nasal tissue specimens at 37°C in different matrices (pH 7.4 and pH 5.5) over a range of incubation periods. The resultant fragments were then separated using a C4 column. Detection and characterization of the fragments were performed by mass spectrometry using total ion current mode. The identities of some major fragments identified in the tissue homogenates were confirmed using commercially available peptides, with respect to their retention time and mass spectra.


Incubation of DYN 1-17 in inflamed nasal biopsies occasioned 22 fragments at pH 5.5 and 14 fragments at pH 7.4. Upon comparison, a similar range of fragments was observed following metabolism of DYN 1-17 in inflamed rat paw and human nasal tissues, suggesting that similar processing enzymes are present in both tissue homogenates. Intriguingly, DYN 3-14 was the most prevalent and stable fragment produced throughout the incubation period, highlighting a rationale for its activity during an inflammatory response.


Differential biotransformation fragments of DYN 1-17 are produced in different inflamed tissues, suggesting that disease pathophysiology may influence the fragmentation patterns of this peptide. The major hydrolysis fragment, DYN 3-14, possesses important roles in immunoregulation and may be developed into a potential immunotherapeutic for CRS.

Keywords: inflammation, dynorphin, biotransformation, chronic rhinosinusitis, immunotherapy

Oral 3.9

Identification of Selective Inhibitors of Phosphodiesterase 4B using Pharmacophore-Based Virtual Screening and Molecular Docking Approaches

Mayasah Al-Nema¹, Anand Gaurav¹*

¹Faculty of Pharmaceutical Sciences, UCSI University, Kuala Lumpur, Malaysia

Corresponding author: Anand Gaurav, anand.pharma@gmail.com


Phosphodiesterase 4B (PDE4B) catalyzes the inactivation of cyclic adenosine monophosphate (cAMP) which is a pivotal second messenger responsible for various biological processes. This enzyme is expressed in immune, inflammatory, and airway smooth muscle cells, thus its inhibition is considered as a treatment option for inflammatory pulmonary disorders i.e. asthma and chronic obstructive pulmonary disease. In contrast to PDE4B, PDE4D is expressed in the area postrema and nucleus of the solitary tract, which are responsible for the emetic response. In this regard, a PDE4B inhibitor selective over PDE4D is expected to be useful anti-inflammatory agent.


A shared feature pharmacophore model for both ligand and structure-based pharmacophore models for PDE4B inhibitors was developed and used as a query in virtual screening of two libraries to identify novel scaffolds that selectively inhibit PDE4B. The hit compounds were filtered according to Lipinski’s rule of five and pharmacophore fit score. Finally, the molecular docking study was applied to evaluate the affinity and selectivity of the hit compounds to PDE4B over PDE4D.


The resulted shared feature pharmacophore model consists of three features: a hydrogen bond acceptor, a hydrophobic unit, and an aromatic ring. The pharmacophore model was used in the virtual screening of Maybridge and SPECS databases. The hit compounds were filtered based on Lipinski’s rule of five and pharmacophore fit score higher than 38.90. Nine compounds were identified as final hits, where four compounds showed higher affinity to PDE4B over PDE4D in molecular docking.  


In this study, four selective PDE4B inhibitors were identified using a combination of ligand-based virtual screening and molecular docking. These inhibitors can be further evaluated for their PDE4B inhibition activity in vitro and anti-inflammatory effect in vivo.

Keywords: Phosphodiesterase 4B, Asthma, Chronic obstructive pulmonary disease, COPD, Pharmacophore, Docking

Oral 3.10

Synthetic Cardamonin Inhibits Migration of Serum-Induced Human Bronchial Smooth Muscle Cells (HBSMCs)

Nazmi Firdaus Musa¹, Daud Ahmad Israf Ali¹*, Tham Chau Ling¹, Hanis Hazeera Harith¹, Manraj Singh Cheema¹

¹Faculty of Medicine and Health Sciences, University Putra Malaysia

Corresponding author: Daud Ahmad Israf Ali, daudaia@upm.edu.my


Increased smooth muscle mass is a prominent features of airways remodelling. This leads to persistent airflow limitation and impaired lung function which contribute to asthma. Inhaled corticosteroid, a primary treatment in asthma management, is unable to inhibit airway remodelling. Hence, effective treatments targeting airway remodelling in asthma are urgently needed. Increased smooth muscle mass may be attributed to both the proliferation and migration of smooth muscle cells. Cardamonin is a chalcone analog that has been reported to inhibit migration of vascular smooth muscle cells. However, its effects upon the migratory capability of bronchial smooth muscle cells have yet to be determined. The ability of synthetic cardamonin to suppress serum-induced Human Bronchial Smooth Muscle Cells (HBSMCs) migration and the mechanisms involved were therefore be evaluated.


Briefly, HBSMCs were treated with the cardamonin (0.75-12.5uM) in a complete media for 48 hours. The effects of cardamonin on HBSMCs migration were assessed through scratch and transwell assays. Meanwhile the effects of cardamonin (12.5uM) on the expression of proteins associated with cell migration (RhoA and phospho-cofillin) were determined by western blot.


The scratch and transwell assays demonstrated that synthetic cardamonin (0.78-12.5uM) reduced the HBSMCs migration in a dose-dependent manner. The inhibitory effect of cardamonin  on migration of serum-induced HBSMCs is associated with a reduced expression of RhoA and increased phosphorylation of cofillin.


Synthetic cardamonin is able to reduce the migration of serum-induced HBSMCs via downregulation of RhoA and upregulation of phosphorylation of cofillin.

Keywords: airway remodelling, smooth muscle migration, serum-induced Human Bronchial Smooth Muscle Cells, cardamonin

Oral 3.11

Anti-inflammatory effect of celery squeezed juice on carrageenan induced rat paw edema

Syarief Hudaya1* and Hardian Hardian2

1Universitas Wahid Hasyim, Indonesia
2Diponegoro University, Indonesia

Corresponding author: Dr. Syarief Hudaya, drsyariefhudaya@yahoo.co.id 


Celery (Apium graveolens L.) contains several active compounds with some anti-inflammatory activities.  The squeezed juice of celery is commonly used in the community to treat various diseases including inflammation. The aim of this study is to determine the anti-inflammatory effect of celery squeeze juice on acute inflammation.


The study design was two-groups pre- and post-test design. Samples were 16 male Wistar rats, age 12 weeks. Rats were randomly allocated into 2 groups: celery group (n=8) received 54 mg squeezed juice of celery per oral; control groups (n=8) received NaCl 0.9% per oral. One hour after celery or NaCl administration, the left paws were injected with carrageenan 1 % dissolved in 0.05 mL NaCl 0.9% intraplantar. Paw volumes were determined by plethysmography at 0 hour (before) until 4 hours with 1 hour interval. Repeated measure ANOVA and unpaired t-test were used for data analysis.


The average paw volume at 0, 1, 2, 3 and 4 hours in celery group were 0.068±0.0085, 0.099±0.0136, 0.078±0.0131, 0.074±0.0115 and 0.073±0.0085 mL respectively (p<0.001); in control group were 0.078±0.0183, 0.100±0.0104, 0.089±0.0125, 0.087±0.0116 and 0.081±0.0099 mL (p<0.001). The differences of paw volume between celery group and control group at 0, 1 and 2 hours were not significantly different (p=0.1, p=0.8 and 0.2 respectively) but at 3 and 4 hours was significantly different (p=0.04 and 0.04 respectively)


Squeezed juice of celery has potency to inhibit acute inflammation

Keywords: celery, Apium graveolens, carrageenan, rat, inflammation, edema

Oral 3.12

Synthesis, in vitro thymidine phosphorylase inhibitory potential and molecular docking study of 3-formylcoumarin analogs

Syed Adnan Ali Shah¹,²*, Muhammad Taha³, Muhammad Afifi¹,², Sadia Sultan¹,²

¹Faculty of Pharmacy, Universiti Tecknologi MARA Puncak Alam Campus, 42300 Bandar Puncak Alam, Selangor D. E., Malaysia
²Atta-ur-Rahman Institute for Natural Product Discovery (AuRIns), Universiti Teknologi MARA (UiTM), Puncak Alam Campus, 42300 Bandar Puncak Alam, Selangor D. E., Malaysia
³Department of Clinical Pharmacy, Institute for Research and Medical Consultations (IRMC), Imam Abdulrahman Bin Faisal University, P.O. Box 1982, Dammam 31441, Saudi Arabia

Corresponding author: Syed Adnan Ali Shah, benzene301@yahoo.com/ syedadnan@salam.uitm.edu.my


Over expression of thymidine phosphorylase (TP) plays a role in the pathogenesis of several conditions such as rheumatoid arthritis, chronic inflammatory diseases, psoriasis and tumor angiogenesis. Hence, finding inhibitors against TP is important. In this regard, a series of seventeen analogs of 3-formylcoumarin (117) were synthesized and screened for thymidine phosphorylase inhibitory potential.


The starting material, 4-chloro-2-oxo-2H-chromene-3-carbaldehyde (1 mmol) was allowed to react with various known benzohydrazides (1 mmol) in the presence of 1 ml acetic acid as catalyst. Methanol (50 ml) was added as the solvent. The reaction was refluxed for 6 h and reaction mixture was checked for completion using TLC. Upon completion, the reaction mixture was rotavapor and residue collected was rinsed with diethyl ether to produce pure products (117). Thymidine phosphorylase/PD-ECGF (human and E. coli) activity was determined by measuring the absorbance at 290 nm spectrophotometrically. AutoDock 4.2 was used to identify the binding modes responsible for the activity.


All analogs showed a variable degree of thymidine phosphorylase inhibition with IC50 values ranging between 0.90 ± 0.01 and 53.50 ± 1.20 M. The standard inhibitor, 7-Deazaxanthine, showed an IC50 value of 38.68 ± 1.12 M. The different IC50 values shown by the analogues can be attributed to the different substituents on the phenyl ring of 3-formylcoumarin. Molecular docking study was carried out to understand the binding interaction of the most active analogs. Docking for the most active derivative, compound 16, showed that it interacts with enzyme through a number of hydrogen bonds and hydrophobic interactions. The docking results also revealed that compound 16 occupies the phosphate binding site with a total of four hydrogen bond interactions.


We have synthesized seventeen analogs of 3-formylcoumarin (117) and screened for thymidine phosphorylase inhibitory activity. Out of seventeen analogs, fifteen analogs, 112 and 1517, showed good inhibition which is many folds better than the standard 7-Deazaxanthine.  Compound 16 exhibited promising inhibitory potential among all analogs. Molecular docking study of compound 16 showed several hydrogen bonds and hydrophobic interactions with thymidine phosphorylase enzyme.

Keywords: 3-formylcoumarin, thymidine phosphorylase inhibition, molecular docking, structure-activity relationship


Oral 4.1

Evaluation of the Anti-Oxidant and Anti-Glycemic Activity of Selected Traditional Medicinal Plants After In Vitro Digestion

Zhi Xiang Ng¹*, Ann Na See ¹

¹School of Biosciences, Faculty of Science, University of Nottingham, Semenyih, Selangor, Malaysia

Corresponding author: Zhi Xiang Ng, ZhiXiang.Ng@nottingham.edu.my


Although there were numerous studies on the therapeutic potential of local traditional medicinal plant (TMP) in South East Asia, relatively few studies have evaluated their bio-accessibility. The changes in the phenolic content, anti-oxidant, and anti-glycemic activities of TMP following the digestion process is yet to be addressed. This study aimed to investigate the effect of in vitro digestion on the anti-oxidant and anti-glycemic activities of twelve selected local TMP.


The moisture content of each TMP was determined before subjected to in vitro oral, gastric and duodenal phases of digestion. The raw and digested TMPs were compared for their anti-oxidant and anti-glycemic activities with seven in vitro mechanism based assays, namely the ferric reducing anti-oxidant power, trolox equivalent anti-oxidant capacity, 2,2-diphenyl-1-picryl-hydrazyl radical scavenging activity, total phenolic and flavonoid content, anti-α glucosidase and anti-α amylase activity.


Centella asiatica showed the highest total anti-oxidant activity (reducing power and radical scavenging), phenolic and flavonoid contents while Morus alba and Dioscorea polystachya possessed the highest anti-α glucosidase and anti-α amylase potential. In vitro digestion significantly (p<0.05) increased the total anti-oxidant activity of four TMPs. Similar trend was observed in the anti-glycemic activity of all TMPs. However, the phenolic and flavonoid contents were significantly (p<0.05) reduced in selected TMPs after the digestion except for Angelica sinensis, Ginkgo biloba and Lycium barbarum. The TMP phenolic content was positively correlated with the total anti-oxidant activity (r=0.840, p<0.001).  Principle component analysis showed that 47.2% variation in the anti-oxidant and anti-glycemic activities of TMP was attributed to the digestion process while 37.3% was due to the different variety of TMP.


In summary, Centella asiatica has the overall highest anti-oxidant index among the twelve TMP. The digestion process could improve the anti-oxidant and anti-glycemic activities of selected TMP, especially Dioscorea polystachya, Ginkgo biloba and Lycium barbarum.

Keywords: Antioxidant, anti-glycemic, in vitro digestion, phenolic content, traditional medicinal plants

Oral 4.2

Nanoemulsified Rice Bran Wax Policosanol Improves Cardiovascular Disease Risk Markers by Regulating Lipid Metabolism in Hyperlipidemic Rats

Aminu Ishaka¹, Mustapha Umar Imam², Maznah Ismail³ and Zayyanu Umar Usman⁴

¹Department of Medical Biochemistry, College of Health Sciences, Usmanu Danfodiyo University, Sokoto, Nigeria
²Precision Nutrition Innovation Institute, College of Public Health, Zhengzhou University, Zhengzhou, 450001, Henan Province, China.
³Laboratory of Molecular Biomedicine, Institute of Bioscience, University Putra Malaysia, 43400 Serdang, Selangor Malaysia
Department of Physiology, College of Health Sciences, Usman Danfodiyo University Sokoto, Nigeria


Policosanol is a long-chain alcohols mixture present in animal and plant waxes. It has been shown to have several biological effects such as lipid-lowering, antiplatelet aggregation, and relief of intermittent claudication. However, it has low bioavailability. We have developed and characterized rice bran wax policosanol nanoemulsion (NPOL) previously. To investigate the effects of NPOL on cardiovascular disease (CVD) risk markers and lipid metabolism, hyperlipidemia was induced in Sprague Dawley rats using high-fat diet containing 2.5% cholesterol. The hyperlipidemic rats were treated with NPOL and rice bran wax policosanol (POL) in comparison with normal diet (ND), high-cholesterol diet (HCD) and simvastatin (SMV)-treated rat groups. Weight, lipid profile, plasma oxidized low-density lipoprotein (ox-LDL), homocysteine and F2-isoprostane were determined. Transcriptional regulation of selected hepatic lipid metabolism genes was also evaluated. NPOL reduced weight gain and improved lipid profile, plasma ox-LDL, homocysteine, F2-isoprostane; and regulated hepatic mRNA expressions of ABCA1, ApoA1, ApoE, and LDLR compared to POL. The results suggest that NPOL could improve CVD risk through ameliorating its markers and modulating lipid metabolism.

Keywords: Cardiovascular disease, Rice bran wax policosanol, nanoemulsion, lipid metabolism.

Oral 4.3

Chronic Administration of Apocynin and Catalase Ameliorates The Blood Pressure and Renal Functional Parameters Attributed By L-NAME Induced Systemic Arterial Hypertension in Wistar-Kyoto Rats: The Role of Oxidative Stress

Tan Yong Chia¹*, Munavvar Zubaid Abdul Sattar¹, Nurzalina Binti Abdul Karim Khan¹, Ashfaq Ahmed², Mohammed Hadi Abdullah³, Ho Yoke Mei¹, Gurjeet Kaur Chatar Singh⁴, Edward James Johns³

¹Cardiovascular and Renal Physiology Research Laboratory, School of Pharmaceutical Sciences, University Sains Malaysia.
²Department of Pharmacology and Toxicology, School of Medicine, Virginia Commonwealth University, Richmond, United States.
³Department of Physiology, University College Cork, Cork, Ireland.
Institute for Molecular Medicine Research, University Sains Malaysia.

Corresponding author: Tan Yong Chia, samual_84@hotmail.com


It is believed that hypertension is associated with increased vascular oxidative stress. There is still a debate whether oxidative stress is a cause or a result of hypertension. Animal studies have generally supported the hypothesis that increased blood pressure is associated with increased oxidative stress. The present study investigates the effect of Apocynin-NADPH oxidase inhibitor and catalase in scavenging the free radical and oxidative stress contributed by L-NAME, a nitric oxide synthase inhibitor, induced hypertension model.  


Forty eight male Wistar-Kyoto rats (200-250g) were randomly assigned into 8 groups (n=6 per group). L-NAME (15mg/kg/day p.o) was used to induce hypertension in selected groups. Apocynin (2.5 mmol/L p.o) and catalase (10000 U•kg-1•day-1, i.p. bolus) were administered into all groups for 14 days. Renal functional parameters and non-invasive blood pressure were studied weekly. Renal hemodynamics parameters such as renal cortical blood perfusion and pulse wave velocity were also recorded. In addition to that, oxidative stress markers i.e. plasma malondialdehyde, superoxide dismutase, nitric oxide and total antioxidant capacity were used to determine the degree of oxidative stress. Kidney samples were used for histopathological studies. All the data were presented in mean±SEM and were subjected to Two-way ANOVA analysis followed by Bonferroni post hoc test with significant level at 5%.


The elevation of blood pressure was observed in L-NAME treated rats on day 7 onwards. Similarly, the deterioration of renal functional parameters and oxidative stress markers were also reported (all P<0.05). Renal histological studies showed that L-NAME treated rats experienced mild arteriolar congestion with minor inflammatory cells in cortical region. Administration of apocynin and catalase reduced blood pressure, improved renal functions and up-regulated the antioxidant defense mechanisms.


This study suggests that apocynin and catalase can be considered as a potential therapeutic option in future hypertension management.  

Keywords: Apocynin, Catalase, L-NAME, Hypertension and Oxidative Stress

Oral 4.4

Obesity, insulin resistance and the rice you eat: Any link?

Bilyaminu Abubakar¹,²*, Norhasnida Zawawi³, Abdul Rahman Omar¹, Maznah Ismail¹

¹Department of Pharmacology and Toxicology, Usmanu Danfodiyo University, Sokoto, Nigeria.
²Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
³Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.

Corresponding author: Bilyaminu Abubakar, abu3163abu@yahoo.co.uk


Type 2 diabetes is a metabolic disorder with established, well-defined precursors. Obesity and insulin resistance are amongst most important factors in predisposition to diabetes. Rice is a staple for about half the global population and its consumption has been strongly linked with diabetogenesis. We assert that tackling the prevalence of predisposing factors by modifying certain rice cultivars could reduce the global burden of obesity and insulin resistance, and by extension type 2 diabetes.


Several rice cultivars with various properties were fed to nulliparous rats (five weeks old at the start of the experiment) for 90 days. They were then returned to a diet of standard pellets and mated with males on a standard diet. The resulting pups and dams were investigated for obesity and insulin resistance markers.


We found that germination caused reduce predisposition to obesity and insulin resistance than high amylose content. The combined reducing effect of germination and high amylose content on predisposition to obesity and insulin resistance was greater than the sum of their independent effects. Polished (white) rice with a low amylose content predisposed dams on a high-fat diet to markers of insulin resistance and obesity and this predisposition was inherited by their F1 offspring.


Overall, the results suggest that harnessing the beneficial properties of germination and amylose in rice would reduce the burden of obesity and insulin resistance, which are known to be key risk factors for development of type 2 diabetes.

Keywords: Obesity, Insulin resistance, Germinated brown rice, amylose content

Oral 4.5

Effects Of Morinda Citrifolia On Diabetes Induced Renal Tissue Apoptosis

Indranila K. Samsuria1* and Hardian Hardian1

1Diponegoro University, Indonesia

Corresponding author: Dr. Indranila K. Samsuria, nila_fkundip@yahoo.com


Diabetic nephropathy (DN) is a microvascular complication due to chronic hyperglycemia. Apoptosis of nephron cells are the major feature of DN. The aim of this study is to investigate the effects of Morinda citrifolia (MC) on nephron cells apoptosis due to DN.


Subjects were 30 male Sprague Dawley rats. All rats were given streptozotocin (STZ) 40 mg/kg BW to induce diabetes. The rats were randomly allocated into 5 groups based on the dose of MC extract/kg BW (MC 10 mg, 20 mg, 40 mg, 80 mg and zero extract acts as a control group). MC extracts were given at 8 weeks after STZ induction, once a day for 2 weeks. Apoptosis index (AI) was measured using TUNEL assay. Caspase-3 activities were measured using immunohistochemistry.


AI of glomerular and tubular cells of MC 20, 40 and 80 mg were significantly lower than control group. AI of glomerular cells in control groups, MC10, MC20, MC40 and MC80 were 9.7±0.44, 8.9±0.89, 4.3±0.31, 6.1± 0.51, and 7.4±0.47, respectively. AI of tubular cells was 9.7± 0.47, 9.3±0.80, 4.6±0.28, 6.3±0.40, 7.6± 0.47 respectively. Caspace-3 expression of glomerular of MC20 and MC40 and tubular cells of MC 20, 40 and 80 mg also significantly lower than control. Caspace-3 expressions of glomerular cells were 3.2±0.46, 2.8±0.43, 1.4±0.27, 2.3±0.33 and 3.2±0.27, respectively. Caspace-3 expressions of tubular cells were 13.0± 0.79, 9.8±1.02, 4.8±0.72, 8.1±0.70 and 10.8±0.2 respectively. The most effective dose MC to inhibit apoptosis of nephron cells was 20 mg/kg BW.


MC extracts have a potency to inhibit nephron cells apoptosis due to DN.

Keywords: Morinda citrifolia, Diabetic Neuropathy, apop´tosis, TUNEL, Caspace-3

Oral 4.6

Changes of ghrelin, insulin and leptin in obese young adult following diet plus exercise

Etika Ratna Noer¹*, Luthfia Dewi¹, Martha Ardiaria¹, Darmawati Ayu Indraswari²

¹Nutrition Department, Diponegoro University, 50275, Indonesia.
²Anatomy and Physiology Department, Diponegoro University, 50275, Indonesia.

Corresponding author: Etika Ratna Noer, etikaratna@fk.undip.ac.id


Weight loss improves ghrelin, insulin, leptin levels and is correlated with energy intake reduction in obese young adult. Thus, diet plus exercise contribute to dependent effect of appetite hormones. This study aimed to examine the effect of diet plus exercise on acyl ghrelin, insulin and leptin levels.


Thirty-three obese young adult with BMI >25 kg/m2, aged (19-20 years) were randomized into two groups: diet plus exercise (n=18) or placebo (n=15). The diet plus exercise program was given for four-weeks. Subjects received isocaloric containing high fiber and aerobic exercise twice a week. Blood samples for acyl ghrelin, insulin, and leptin analysis were obtained pre-post intervention. The biomarkers were assessed by enzyme-linked immunosorbent assay.


The diet plus exercise group (DEG) showed a significantly decrease in acyl ghrelin and insulin (-93.08±171.73 pg/mL; -9.72±14.75 ng/mL, P<0.001) compared to control group (CG) (-15.55±27.49 pg/mL; -0.37±8.70 pg/mL, P<0.042). Furthermore, both groups revealed an increase of leptin (14.69±27.87 pg/mL for DEG; 5.35±37.98 pg/mL for CG).


Diet plus exercise promote weight loss in obese young adult by improving appetite hormones.

Keywords: acyl ghrelin, insulin, leptin, diet-exercise, obese

Oral 4.7

Aqueous extract of Garcinia atroviridis fruit improves oxidative status of high fat diet-induced hyperlipidemic rats

Majed A. Al-Mansoub¹*, Mohd. Z. Asmawi¹ and Vikneswaran Murugaiyah¹*

¹Department of Pharmacology, School of Pharmaceutical Sciences, University of Science Malaysia, Malaysia

Corresponding authors: Dr. Majed A. Al-Mansoub, majed@usm.my and Dr. Vikneswaran Murugaiyah, vicky@usm.my

Garcinia atroviridis, is a well-known plant in South East Asia, commonly used in cooking and traditional medicine. The present study was undertaken to evaluate the effect of aqueous extract of ripe fruits (AqE-RFS) on the oxidative status of high-fat diet (HFD) induced hyperlipidemic rats.

Male Sprague-Dawley (SD) rats are divided into six groups of 6 rats per group. Control group rats received normal control diet for a period of six weeks. The rest of animals were fed with HFD and simultaneously treated orally with different doses of AqE-RFS (250, 500 and 1000mg/kg body weight/day).

Oral administration of the AqE-RFS for six weeks significantly reduced the liver and serum lipid peroxidation by decreasing malondialdehyde (MDA) levels at both doses of 500mg/kg (p<0.05) and 1000mg/kg (p<0.01) when compared to HFD control rats. The levels of hepatic glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) were significantly (p<0.01) increased in animals treated with AqE-RFS at 1000mg/kg. Similarly, in serum of HFD treated rats with 1000mg/kg, there was a significant increase (p<0.001) in GSH and SOD levels as well as significant in CAT, GPx and GR levels (p<0.01) as compared to the HFD rats. Strong negative correlations were observed between hepatic and serum MDA contents and antioxidant markers.

Administration of AqE-RFS attenuates the oxidative stress and increased the antioxidant enzymes in a dose-dependent manner. The findings revealed that AqE-RFS has potential protective effect against oxidative damage in HFD-induced hyperlipidemic rats.

Keywords: Garcinia atroviridis, high-fat diet, Hyperlipidemia, Lipid Peroxidation, antioxidant

Oral 4.8

Tissue-specific regulation of insulin-PI3K/Akt, KEAP1-NRF2 and PKC/NF-κB signaling in high fat diet-fed and nicotinamide/streptozotocin-induced diabetic rats

Der Jiun Ooi¹,²* and Maznah Ismail²

¹Faculty of Dentistry, Mahsa University, Malaysia
²Putra Malaysia University, Malaysia

Corresponding author: Dr. Der Jiun Ooi, ooiderjiun@gmail.com


Type 2 diabetes mellitus (T2DM), a heterogeneous metabolic disorder, is due primarily to complex interplay of multiple inherited and acquired factors that adversely impair glucose metabolism. In particular, impaired insulin actions at multiple target tissues lead to eccentric glucose, lipid and redox homeostasis. While it is widely recognized that insulin resistance underlies the pathophysiology of T2DM, the complicated tissue-specific gene expression regulation and phenotypic association of the disease, particularly in diabetic animal model, remain largely under-studied. The present study aims to determine the correlated tissue-specific gene expression changes in experimental diabetic rat model.


The high fat diet-fed and nicotinamide/streptozotocin-induced mild diabetic (DIA) rats were used in the present study. Normal diet- (NOD) and high fat diet-fed (HFD) rats served as controls. The expressions of key genes involved in insulin-PI3K/Akt, KEAP1-NRF2 and PKC/NF-κB signaling, as well as correlation between gene expression changes, were profiled from white adipose, skeletal muscle, and liver tissues. The differential and non-differential molecular network interactions were further identified.


Consistent with previous literature, statistical analyses revealed severe inflammation and perturbed metabolic functions in both HFD and DIA groups. Interestingly, potential tissue-specific compensatory antioxidant mechanisms were also observed across all the target tissues. Further dynamical analyses on differential and non-differential expression networks demonstrated tissue-specific modulations of inflammatory and antioxidant responses in relation to deranged insulin signaling.


The present study suggested tissue-specific and inter-tissue gene expression interactions in the mild diabetic rat model. Further studies and association with tissue-specific gene expression changes in human diabetic patients may further validate the use of present animal model to be verified fit for diabetes translational research purpose.

Keywords: type 2 diabetes mellitus, tissue-specific gene expression, insulin-PI3K/Akt signaling, KEAP1-NRF2 signaling, PKC/NF-κB signaling

Oral 4.9

Isolation of Scopoletin from Paederia foetida and its Antidiabetic Potential Using In silico Model

Dai Chuan Tan¹*, Ku Idayu Idris¹, Nur Kartinee Kassim¹*, Pei Cee Lim¹, Intan Safinar Ismail¹, Muhajir Hamid¹ and Rou Chian Ng¹

¹Universiti Putra Malaysia , Malaysia

Corresponding authors: Dai Chuan Tan, tandaichuan@yahoo.com and Dr. Nur Kartinee Kassim, kartinee@upm.edu.my


Paederia foetida L. (Rubiaceae) is an edible plant widely distributed in India, Japan, Malaysia, and other Asian countries. This study investigates the enzyme inhibition and of P. foetida twig extracts obtained from two different locations namely Johor (PFJ) and Pahang (PFP) and its chemical constituents.


The isolation of phytoconstituents using column chromatography methods and in silico study of isolated compound with receptor (α-glucosidase and α-amylase) was carried out using molecular docking.


The IC50 of α-glucosidase and α-amylase inhibition activity of the PFJ chloroform extract were 245.6 ± 0.01 and 9.595 ± 0.01 µg/mL, respectively. The IC50 of α-glucosidase and α-alpha amylase inhibition activity of PFP chloroform extract were 2457.2 ± 0.01 and 14.83 ± 0.05 µg/mL, respectively. Separation of PFP chloroform extract afforded ergost-5-en-3-ol (1), stigmasterol (2), γ-sitosterol (3) and scopoletin (4) meanwhile PFJ chloroform extract gave (2) and (4). Scopoletin was isolated for the first time from this species. In silico molecular interaction study between scopoletin and α-glucosidase and α-amylase showed that α-glucosidase-scopoletin inhibitor complex had better binding affinity with less energy value (-5.4 kcal/mol) than α-amylase-scopoletin inhibitor complex (-4.9 kcal/mol).


PFJ chloroform extract showed potent inhibitory activity toward α-glucosidase. The in silico assessment of scopoletin as antidiabetic agent indicated that α-glucosidase-scopoletin complex has good stability with binding affinity of -5.4 kcal/mol. P. foetida can be considered as potential resources for natural herbal antidiabetic drugs which can serve as alternative to synthetic oral hypoglycemic drugs with minimum side effects.

Keywords: Paederia foetida, Scopoletin, alpha-Amylase, Alpha-glucosidase, molecular docking

Oral 4.10

Potential Clinically Significant Drug and Gene Interactions Involving Cytochrome P450 Family 2 Subfamily D Member 6 (CYP2D6) Relevant to Opioids Used for Chronic Pain in Community-dwelling Older Australians

Thilani H. Dias¹, Mohitosh Biswas¹, Nilofar Daneshi¹, Elizabeth Holliday², Stephen Hancock², Karen P. Kerr¹, Irene Munro¹, John Attia¹, Rodney Scott¹ and Liz Milward¹*

¹School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Australia
²School of Medicine and Public Health, Faculty of Health and Medicine, University of Newcastle, Australia

Corresponding author: Dr. Liz Milward, liz.milward@newcastle.edu.au


Certain clinically significant genetic variants requiring change of drug or dose or monitoring can alter CYP2D6 enzyme activity in opioid metabolism, constituting a simple drug-gene interaction (simple DGI). Drug-drug interactions (DDIs) involving concomitant use of CYP2D6 inhibitors can also alter opioid efficacy. Hence investigation of DGIs or DDIs or combinations of these (multifactorial DGIs) may help optimise patient responses to opioids This study used genotype data previously obtained in a community study of older Australians to investigate poor or intermediate metaboliser genotypes relevant to opioid therapeutic failure.


Prevalence of selected CYP2D6 poor or intermediate genotypes (DGIs) for opioids of interest (codeine, tramadol, oxycodone) were assessed for 2121 older Australians from the Hunter community study (55 years or older), using Affymetrix Kaiser Axiom arrays or imputed from reference panels. Data for other genotypes were not available. Potential DDIs were assessed from self-reported medication data.


Approximately 17% of participants (95% CI:15%-18%) had the selected clinically significant poor or intermediate metaboliser genotypes. Approximately 23% (95% CI: 15%-31%) of all codeine users (n=96) were at risk of at least one clinically significant DDI involving CYP2D6 inhibitors. Approximately 15% (95% CI:5%-24%) of the 55 participants taking codeine with genotype data, but no participants taking oxycodone or tramadol, had the genotypes investigated, with ~13% (7/55, 95% CI:4%-22%) and ~2% (1/55, 95% CI:-2%-5%) at risk of simple or multifactorial DGIs respectively.


Approximately 1 in 6 participants had clinically significant genotypes relevant to codeine, with over 10% of codeine users having relevant clinically significant DGIs or DDIs. Considering use of non-opioid medications or morphine may improve therapeutic outcomes for such patients. Future studies are planned to investigate ultra-rapid metaboliser genotypes that may increase opioid toxicity.

Keywords: precision medicine, pharmacogenomics, CYP2D6, opioid metabolism, community study


Oral 5.1

The Effect of Listening to the Quran Surah Al-Mulk with Translation on Anxiety and Sleep Quality in Medical Students

Cindar F. Sari¹*, Hana Pertiwi¹, Darmawati A. Indraswari² and Yuriz Bakhtiar²

¹Faculty of Medicine, Diponegoro University, Indonesia
²Department of Physiology, Faculty of Medicine, Diponegoro University, Indonesia

Corresponding author: Cindar F. Sari, cindarfatiha@gmail.com


Reciting Quran, the holy book of Islam or listening to its recitation has been demonstrated worldwide to improve psychological and physiological conditions. Al-Mulk (sovereignty, kingdom) is one of its Surah with supposed benefits. Listening to the Arabic recitation continued with understanding the meaning may result in neural and hormonal enhancement to reduce anxiety and improve sleep quality. This study is to determine the effect of listening to the Quran Surah Al-Mulk on anxiety and sleep quality in medical students.


This is an experimental study with pre- and post-test design. Forty nine medical students of various years were included in the study. The first group of 27 students, the group 1, filled the Hamilton Anxiety Rating Scale (HARS). While the rest 22 students as the group 2 filled the Pittsburgh Sleep Quality Index (PSQI).. The quran recitation, Surah Al Mulk (30 verses) in Arabic followed by Indonesian translation was played using MP3 player once daily for 14 days to both groups. Posttest for both anxiety and sleep quality were applied after the interventions. The Wilcoxon test was used to determine the differences between HARS scores or the PSQI score before and after listening to Al-Mulk and its translation in each group respectively.


There was a significant decrease in HARS score after listening to Al-Mulk with translation in group 1 (p<0.001) compared to before listening. There was also a significant decrease in PSQI in group 2 after listening (p<0.05) compared to before listening.


Listening to Quran Surah Al-Mulk and its translation reduces anxiety and sleep quality in medical students.

Keywords: quran, anxiety, sleep quality, medical students, HARS, PSQI

Oral 5.2

The effect of zinc supplementation on the improvement of clinical symptoms and the quality of life of persistent moderate severe allergic rhinitis patients

Anna M. Dewi1*, Dian I. Setyorini1 and Suprihati1

¹Diponegoro University, Indonesia

Corresponding author: Dr. Anna M. Dewi, anna_drtht@fk.undip.ac.id


The aim of Allergic Rhinitis (AR) therapy is to overcome the symptoms in the early and late phases allergic reactions. The ARIA 2008 guideline recommends intranasal corticosteroids and antihistamine for 2 to 4 weeks for persistent moderate severe AR patients. Zinc as an anti-inflammatory is expected to be an alternative for intranasal corticosteroid replacement therapy. The aims of this study is to determine the effect of zinc supplementation on the improvement of clinical symptoms and the quality of life of persistent moderate severe AR patients.


It was an interventional study with pretest and posttest design. Administration of 40mg/day zinc tablets in patients with persistent moderate severe AR treated with cetirizine (treatment group) was compared with cetirizine only (control group). Assessment was done by Total Symptom Score and Quality of Life Score questionnaires at the beginning of therapy and second week after therapy. Statistical tests used were t test, Wilcoxon, and Mann Whitney test.


There were 34 persistent moderate severe AR patients. Administration of zinc 40 mg/day for two weeks showed a significant reduction of mean total symptom score (treatment group 2,65 ± 2,499 and control group 5,12 ± 2,956, p: 0,012) and mean quality of life score (treatment group 9,12 ± 11,230 and control group 24,24 ± 14,237, p: 0,002).


Zinc supplements is useful to improve clinical symptoms and quality of life for moderate severe persistent AR patients.

Keywords: zinc supplementation, Moderate Severe Persistent, allergic rhinitis, quality of life score (QOLS), Total symptom score

Oral 5.3

Evaluation of cellular antioxidant activity and chemical constituents of Melicope latifolia (DC.) T. G. Hartley

Pei Cee Lim1, Nur Kartinee Kassim1*, Zulfiqar Ali2*, Ikhlas Khan2, 3, Shabana I. Khan2, 3, Khozirah Shaari1 and Amin Ismail4

1Department of Chemistry, Faculty of Science, Putra Malaysia University, Malaysia
2National Center for Natural Products Research, School of Pharmacy, University of Mississippi, United States
3Division of Pharmacognosy, Department of BioMolecular Sciences, School of Pharmacy, University of Mississippi, United States
4Department of Nutrition and Dietetics, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia

Corresponding author: Dr. Nur Kartinee Kassim, kartinee@upm.edu.my


Melicope latifolia (DC.) T. G. Hartley, a Rutaceae family species, known as ‘kisampang’ and ‘pepau’ are distributed in Southeast Asia – Malaysia, Indonesia, Philippines to Papua New Guinea in the Pacific. Traditionally, Melicope latifolia leaves are used as remedy for fever and cramps. In Indonesia, the leaves are used in folk medicine to cure jaundice and malaria. Other Melicope species were reported to possess antiviral, anti-inflammatory and antioxidant activities. However, to date, there are limited reports on chemical constituents and biological potential of this species.


Powdered leaves of Melicope latifolia were extracted successively using hexanes, ethyl acetate and methanol. Total phenolic contents (TPC) and antioxidant capacities namely 2,2ʹ-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, oxygen radical absorbance capacity (ORAC) and copper reducing antioxidant capacity assay (CUPRAC) of the extracts were evaluated. Phytochemical study on the methanol extract yielded five flavonoids and flavonoid glycosides which were then subjected to cellular antioxidant activity (CAA) assay.


Melicope latifolia methanol extract exhibited the highest phenolic content with 57mg GAE/g extract compared to other extracts. Maximum DPPH scavenging activity and ORAC values were also observed in methanol extract with 70mg TE/g extract and 1830µmol TE/g extract respectively. The methanol extract also possessed significant reducing ability on CUPRAC with the value of 187mg TE/g extract. Chemical constituents named taxifolin, quercetin 3-O-robinobioside, kaempferol 3-O-rutinoside, kaempferol 3-O-glucoside and kaempferol 3-O-arabinoside were isolated for the first time from this species. These compounds were subjected to CAA assay. Taxifolin was the most effective in decreasing the cellular oxidative stress compared to other compounds.


Methanol extract of Melicope latifolia appears to have the most significant antioxidant capacity. Isolated phenolic compounds from the extract certainly contribute to its anti-oxidative properties to some extent.

Keywords: Rutaceae, Flavonoid, Melicope latifolia, Flavonoid glycosides, antioxidant

Oral 5.4

Role of red fruit (pandanus conoideus) towards photoaging inhibition in mice skin

Renni Yuniati1*, Sylvia C. Sugianto2, Saekhol Bakri3, Dwi Retnoningrum4 and Farmaditya Mundhofir5
1Department of Dermatology and Venereology, Diponegoro University, Indonesia
2Diponegoro University, Indonesia
3Department of Public Health, Diponegoro University, Indonesia
4Department of Clinical Pathology, Diponegoro University, Indonesia
5Department of Histology, Diponegoro University, Indonesia

Corresponding author: Renni Yuniati, renniyuniati@yahoo.com


Early aging is caused by sunlight exposure and can be prevented by antioxidant administration. Lately, people tend to use natural ingredients that contain antioxidants. Red fruit from Papua contains a lot of antioxidants and can be used as one of the alternatives to prevent early aging. This study aimed to know the role of red fruit towards photoaging inhibition, which was assessed from total number of type I procollagen in mice skin.


This was an animal experimental study, mice was used as the model animal and was divided into 3 groups. Group 1 was control group, group 2 was exposed with ultraviolet ray from artificial source, group 3 was exposed with ultraviolet ray and recieved red fruit orally. Biopsy of the skin was done after treatment, checked using immunohistochemistry with staining of type I procollagen and then evaluated using computer program.


This study showed an increased expression of type I procollagen in group 3 compared with group 2.


Red fruit can be used to prevent photoaging.

Oral 5.5

A novel modifier gene of the p53-mediated nucleolar stress response activated by ribosomal protein depletion

Mei S. Wong1, 2, 3, Sheren J. Al-Obaidi1, 2, Katherine M. Hannan1, 2, 3, Nadine Hein1, 2, Kaylene J. Simpson3, 4, Richard B. Pearson2, 3, Amee J. George1, 2, 5* and Ross D. Hannan1, 2, 3, 5
1ACRF Department of Cancer Biology and Therapeutics, John Curtin School of Medical Research, Australian National University, Australia
2Oncogenic Signalling and Growth Control Program, Peter MacCallum Cancer Centre, Australia
3Sir Peter MacCallum Department of Oncology, The University of Melbourne, Australia
4Victorian Centre for Functional Genomics, Peter MacCallum Cancer Centre, Australia
5ANU Centre for Therapeutic Discovery, John Curtin School of Medical Research, Australian National University, Australia

Corresponding author: Dr. Amee J. George, amee.george@anu.edu.au


The nucleolar stress response (NSR) is an important mechanism for maintenance of cellular integrity, which has been implicated in human diseases, particularly in cancer and ribosomopathies (e.g. Diamond Blackfan Anaemia, DBA). Perturbations in ribosome biogenesis (e.g. due to ribosome insufficiency, DNA damage) can activate NSR, where by ribosomal proteins (e.g. RPL5 and RPL11) bind to and inhibit MDM2, thus preventing p53 degradation and causing accumulation of p53. The activated p53 subsequently induces cell cycle arrest, senescence or cell death depending on the cellular background. However, the precise molecular mechanism underlying this process is not well defined and warrants further investigation.


We performed a genome-wide loss of function (RNAi) screen to identify novel modulators of the p53-mediated NSR induced by ribosomal protein insufficiency (RPS19 depletion) and further interrogated a number of these candidates to verify their importance in this process.


Validation of the screening data yielded a selection of candidates that were ranked as high, medium and low confidence; a selection of the high and medium confidence candidates were further validated using in vitro assays. A number of our high confidence candidates were observed to block p53 stabilisation induced by RPS19 knockdown (which are potential therapeutic targets for the treatment of DBA), while a number also enhance p53 stabilisation, which could be utilised to enhance the NSR response (could be used as the basis of p53 induction in cancer therapy). In particular, we have identified a novel NSR modifier that is required for ribosome biogenesis and our current work is to investigate the specific mechanism by which this candidate acts to modulate the NSR.


High throughput functional screening has identified a selection of candidates which modulate the p53-dependent NSR, and these candidates could serve as potential therapeutic targets for the treatment of diseases including ribosomopathies and cancer.

Keywords: High throughput screening, nucleolar stress response, Ribosomal protein (RP)-p53 pathway, Ribosome biogenesis, therapeutic targets

Oral 5.6

Chemical and molecular diversity analysis of Curcuma longa L. across the Indian subcontinent

KAMRAN ASHRAF1, 2, 3* and Mohammad Mujeeb3
1Department of Pharmacology & Chemistry, Faculty of Pharmacy, Universiti Teknologi MARA Puncak Alam, Malaysia
2Atta-ur-Rahman Institute for Natural Product Discovey, Universiti Teknologi MARA, Malaysia
3Bioactive Natural Product Laboratory, School of Pharmaceutical Education & Research, Jamia Hamdard University, India

Corresponding author:  Dr. KAMRAN ASHRAF, kamran1368@puncakalam.uitm.edu.my


Curcuma longa (turmeric) is a valued medicinal plant belonging to the family Zingiberaceae which comprises more than 80 species of rhizomatous perennial herbs and has extensive occurrence in the tropics of Asia, Africa and Australia. Previous studies of turmeric did not show any data of molecular and chemo profiling side by side. As this study could generate good information for better reproducibility and cultivation of this plant. So the aim of the research was to find out chemical and molecular diversity among accessions of turmeric collected across the Indian subcontinent.


Phytochemical constituent curcuminoids of eight accessions of turmeric were evaluated by ultra-performance liquid chromatography – tandem mass spectrometer (UPLC-MS/MS) method. Molecular diversity analysis was done by polymerase chain reaction (PCR) based random amplification polymorphism DNA(RAPD) method.


Results of chemo profiling by ultra-performance liquid chromatography – tandem mass spectrometer (UPLC-MS/MS) showed large variations in the content of curcuminoids (1.408 – 5.027% w/w) and RAPD based molecular diversity analysis also showed excellent genetic variations (44.44% to 100%) among samples of C. longa. Results also showed that Erode (southern part) of India produced higher yield of curcuminoids (50.27 mg/g). These results revealed that C. longa has undergone both chemical as well as genetic variations.


From these results it is concluded that chemical and molecular diversity occurred in C. longa and Erode (southern province) contains the highest quantity of curcuminoids. Erode variety could be superior and can be used for large scale production and cultivation. These variations in turmeric may be due to wide range of ecological conditions within distribution area of its population in India. These outcomes would provide an important input into determining efficient management strategies for the cultivation and improvement program of C. longaglobally and in India in particular.

Keywords: Curcuma longa, turmeric, UPLC-MS/MS, RAPD, India

Oral 5.7

Molecular Characterization of blaTEM and blaCTX-M in ESBLs producing Escherichia coli and Salmonella spp. in Poultry farm and Klebsiella pneumoniae and Proteus spp. in Klang River in Malaysia

Nagaraja Suryadevara1*, Kai Boon Yong1, Sharanya Rajan1, Yoke Ing Kwan1 and Michelle Fong Ting Lim1
1Mahsa University, Malaysia

Corresponding author: Dr. Nagaraja Suryadevara, nagaraja@mahsa.edu.my


Multidrug resistance bacteria (MDR) has emerged as a public health threat nowadays. High level of resistance to beta-lactam antibiotics among Enterobacteriaceae was conferred by Extended Spectrum Beta-Lactamase (ESBL) in the poultry farm. The waste material of the livestock could pollute the river, leading to the aquatic environment as a reservoir of antimicrobial resistance.


The physiochemical properties of water samples were determined. Gram-negative microorganisms were identified using differential media and biochemical testing. Confirmed bacterial isolates were tested for multidrug resistance by using Kirby Bauer method. The ESBL strains were determined phenotypically using Combination Disc Test (CDT) and Double Disc Synergy Test (DDST). Bacterial genomic DNA was extracted from positive ESBL strains, blaTEM and blaCTX-M genes were amplified using Polymerase Chain Reaction (PCR). The amplified products were sequenced to identify the mutated genes in MDR strains.


Out of 88 samples collected from poultry farm, 73 (83.0%) E.coli and 59 (67.0%) Salmonella spp. were identified. Of 50 water samples from Klang River, 31 (62.0%) Klebsiella pneumoniae and 13 (26.0%) Proteus spp. were found. In total, 11 (15.1%) E.coli, 14 (23.7%) Salmonella spp., 12 (38.7%) Klebsiella pneumoniae, and 7 (53.8%) Proteus spp. were MDR strains. From there, 8 (72.7%) E.coli, 7 (50.0%) Salmonella spp., 5 (16.1%) Klebsiella pneumoniae and 5 (38.5%) Proteus spp. were ESBL positive strains. The genes blaTEM and blaCTX-M were seen exhibited in 3 (42.9%) and 7 (87.5%) E.coli , 1(14.3%) and 5(71.4%) Salmonella spp., 1 (20.0%) and 4(80.0%) Klebsiella pneumoniae and 1 (20.0%) and 4 (80.0%) Proteus spp., respectively.


Classic blaTEM and blaCTX-M genes were present in E.coli and Salmonella spp. from poultry samples; and Klebsiella pneumoniae and Proteus spp. from river water samples. However, blaCTX-M gene was seen more prominent compared to blaTEM gene in the isolated ESBL strains.

Keywords: MDR – (multi drug resistant), ESBL – Extended-spectrum beta-lactamase, Bla TEM, bla CTX -M1, DDST, CDT

Oral 5.8

Quantitative analysis of andrographolide, a biomarker of Andrographis paniculata, using qNMR spectroscopy

Sreenivasa Rao Sagineedu1*, Ka Ling Kong2, Sze Ying Phang2, Boon Ling Toh2, Vun San Chu2, Ahmad Sazali Hamzah3 and Johnson Stanslas4
1Department of Pharmaceutical Chemistry, School of Pharmacy, International Medical University, Bukit Jalil, 57000, Malaysia
2School of Pharmacy, International Medical University, Bukit Jalil, 57000, Malaysia
3Organic Synthesis Laboratory, Institute of Science, Universiti Teknologi MARA (UiTM), 40450, Malaysia
4Department of Medicine, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400, Malaysia


Corresponding author: Dr. Sreenivasa Rao Sagineedu, sreenivasarao@imu.edu.my


Andrographis paniculata also known as “king of bitters”, is a well-known traditional medicinal herb which has been widely used in Asian countries. Andrographolide (AGP) is the major bioactive constituent of A. paniculata and a potent biomarker which has been shown to have a wide range of pharmacological properties including anti-inflammatory, antitumor and hypoglycemic activities. In this study, 1H NMR spectroscopy has been successfully introduced to quantify the amount of AGP present in herbal preparations.


1H NMR based quantification approach was established via optimisation of experimental conditions, including NMR pulse sequence and NMR relaxation delay time. The NMR spectra were obtained in dimethylsulfoxide-d6 using maleic acid as the internal standard.


The results indicate that qNMR spectroscopy is a powerful analytical tool for rapid and accurate determination of AGP in herbal preparations. The developed method was found to be linear (r2 = 0.9983) and the percentage recovery for the accuracy was 98.88% – 99.38%. The %RSD for precision, robustness and stability studies was found to be less than 2%. The method has also been successfully applied to test few commercial preparations of A. paniculata sold in the local market.


The qNMR spectroscopy is found to be a rapid and versatile tool for the qualitative and quantitative determination of AGP as well as impurities in herbal preparations.

Keywords: Andrographis panicualta, andrographolide, quantification, NMR, Herbal

Oral 5.9

The Effect Of Autonomous Sensory Meridian Response Stimulation On Sleep Quality

Sari S. Febriani1, Hardian Hardian1* and Tanjung A. Sumekar1
1Diponegoro University, Indonesia

Corresponding author:  Dr. Hardian Hardian, dokterhardian@gmail.com


Sleep is time for brain to recover of biochemistry and physiologic function of the brain after standby during day time. Stress may cause sleeping disturbance and reduce sleep quality. Stimulus Autonomous Sensory Meridian Response (ASMR) can create a feeling of relaxation and comfort that helps one to fall asleep. The aim of this study is to determine the effect of ASMR on sleep quality.


Study subjects were undergraduate students of Medical Faculty Diponegoro University academic year 2017 (n=30). Subjects were randomly allocated into two groups. Group I was given ASMR stimulation (n = 15) and group 2 was not given any treatment (n = 15). ASMR stimulation was delivered by watching a 15 minutes of ASMR video from a laptop before going to sleep at night. The stimulation were given for 7 consecutive days.
Sleep quality was measured using the Pittsburgh Sleep Quality Index (PSQI) questionnaire. Measurements are made before receiving the stimulus and after receiving the stimulus. Data was analysed using Mann Whitney test and Wilcoxon test.


In Group I, PSQI score before ASMR stimulation was 7.0 ± 1.69 and after was significantly reduced to 3.9 ± 0.9 (p<0.001). In Group II, before was 6.7 ± 1.45 and after was increased to 6.8 ± 1.61(p=0.6). PSQI score before of Group I was not significantly different than group II (p=0.6). PSQI score after of Group I was significantly lower than Group II (p<0.001)


ASMR stimulation can improve sleep quality.

Keywords: ASMR, PSQI (Pitzburg Sleep Quality Index), Sleep, nervous, video

Oral 5.10

Sodium Caseinate Nanomicelles as a Novel Drug Delivery System for Doxorubicin

Farah Rehan1*, Manish Gupta1* and Nafees Ahemad1
1Monash University Malaysia, Malaysia

Corresponding author: Ms. Farah Rehan,  farah.rehan@monash.edu
Dr. Manish Gupta, manish.gupta@monash.edu
Dr. Nafees Ahemad, Nafees.Ahemad@monash.edu


Breast cancer is the most common cancer amongst women in both the developed and less developed world. It impacts over 1.5 million women worldwide each year. Doxorubicin (DOX) is widely used drug for breast cancer, however, researchers are still looking for appropriate delivery system that can overcome acquired resistance and toxicity associated with DOX. Casein nanomicelles, the major fraction of milk protein, are emerging as a novel drug delivery system owing to their various structural and functional properties.


Doxorubicin (DOX) was successfully loaded into sodium caseinate nanomicelles (NaCNs) which were formulated and optimised through magnetic stirring. Zeta size and zetapotential of formulations were measured and then formulations were further characterized through FESEM, TEM, FTIR and XRD. In-vitro drug release study was also assessed by using dialysis membrane.


NaCNs yielded sizes that ranged from 348.8 nm (blank micelles) to 604 nm (drug-loaded micelles) and, negative zeta potential of -22.1 (blank micelles) to -21.9 mV (drug-loaded micelles). DOX showed significant encapsulation efficiency (71.57% ± 4.855) and drug loading (2.01±0.095). Morphological characterisation through FESEM, TEM revealed the spherical morphology of micelles of both unloaded and loaded micelles. XRD data confirmed the amorphous nature of the formulated drug-loaded micelles. FTIR spectra indicated the proper entrapment of the drug and confirmed no chemical interaction between the drug and the micelles. Drug release profile of the loaded micelles revealed that the colloidal drug carrier could release the drug in a slow manner under normal physiological conditions.


The present findings implied the potential of overcoming the limitations associated with DOX by formulating DOX-loaded NaCNs against breast cancer.

Keywords: breast cancer, Nanomicelles, Doxorubicin, FESEM, Encapsulation efficiency

Oral 5.11

Evaluation of lung cancer awareness among pharmacy students in Malaysia: a cross-sectional study

Sohail Ahmad1*, Muhammad Qamar1, Wong Y. Zhi1 and Nurul I. Hassan Bashri1
1Faculty of Pharmacy, Mahsa University, Malaysia

Corresponding author: Dr. Sohail Ahmad, sohail@mahsa.edu.my


Cigarette smoking at an early age and the increasing trend of using e-cigarettes and vaping devices are potential risk factors for developing lung cancer among Malaysians. It accounts for 19.8% of all cancer mortality and is the third most common cancer in Malaysia. This study was conducted to assess the awareness of lung cancer among pharmacy students in Malaysia.


This cross-sectional study recruited a total of 225 pharmacy students enrolled at three private universities situated in Selangor, Malaysia. Post-approval from Cancer Research UK, a validated Cancer Awareness Measure (CAM) was used. CAM consisted of five main sections: socio-demographic data (n=6), level of confidence (n=1), lung cancer age (n=1), warning signs (n=14), and risk factors (n=9) of lung cancer. The data were extracted from completed questionnaires and analyzed descriptively and inferentially using SPSS® version 22.


Majority of the enrolled students were females (75.5%), and Malay (40.8%). The descriptive analyses of correct responses suggested that the students possessed a high level of awareness regarding warning signs (n=139, 61.7%) and risk factors (n=145, 64.4%) of lung cancer. Chi-square test showed that the age categories exhibited statistically significant associations with warning signs, χ2 (4)=14.256, p=0.007, as well as with risk factors, χ2 (4)=13.431, p=0.010. In addition, educational level, χ2 (4)=11.396, p=0.022, and smoking status, χ2 (8)=22.251, p=0.044, showed statistically significant associations with risk factors. The students’ score of warning signs and risk factors were positively correlated as suggested by Spearman’s correlation (r=0.42, p=0.001).


The pharmacy students in Malaysia possess high level of lung cancer awareness. Pharmacy students therefore can contribute actively in community-based lung cancer awareness campaigns after adequate training and under the supervision of healthcare professionals.

Keywords: lung cancer, Malaysian, Awareness, Students, Pharmacy

Oral 5.12

Apoptotic Evaluation and Cytotoxic Effects of Doxorubicin Conjugated CaCO3-nanoparticles on Breast Cancer Cell-line

Hamidu Ahmed1*, Mokrish Ajat2, Rozaihan Mansor2, Intan Shameha Abdul Razak2, Abubakar Danmaigoro2, Alhaji Zubair Jaji2, Jaafaru Sani Mohammed3 and Md Zuki Abu Bakar2*
1Laboratory of Molecular Biomedicine, Institute of Bioscience, University Putra Malaysia, Malaysia
2Faculty of Veterinary Medicine, Putra Malaysia University, Malaysia
3MAKNA Cancer Research, Institute of Bioscience, University Putra Malaysia, Malaysia


Corresponding author: Mr. Hamidu Ahmed, boderel@yahoo.com

Prof. Md Zuki Abu Bakar, zuki@upm.edu.my


Breast cancer is the most common malignant disease which is a serious burden to women worldwide. Aragonite calcium carbonate Nanoparticles loaded with doxorubicin (DOX) has recently emerged as an effective treatment of breast cancer. The aim of this study was to evaluate the efficacy of Doxorubicin-Calcium Carbonate Nanoparticles (DOX-Ar-CC-NPs) on MCF-7 breast cancer cells and to elucidate the mechanism of action, aiming at giving an insight on how DOX-Ar-CC-NPs influence chemo-resistivity and drug target delivery limiting undesirable side effects.


DOX-Ar-CC-NPs was synthesized using novel biomolecules from cheaply available natural sea water Cockle shells and characterized for particle geometry using electron microscopy. The anticancer activity of DOX-Ar-CC-NPs was determined using superoxide dismutase commercial ELISA kit for cell membrane integrity, flow cytometry, fluorescent imaging and electron microscopy for programmed cells death evaluation.


Treatment of MCF-7 cells with DOX-Ar-CC-NPs and DOX showed a dose dependent effect on cell viability. The DOX-loaded-Ar-CC-NPs had significant inhibitory effect on cell viability compared to DOX alone (p<0.05). Similar trend was noticed in cellular apoptosis, oxidative stress markers and cellular uptake evaluation. In addition, the results clearly showed that DOX-Ar-CC-NPs induced necrosis through endocytosis. However, treatment with DOX-Ar-CC-NPs significantly decreased the elevated level of superoxide dismutase 2 (SOD2) compared to untreated MCF-7 cells (control group). Furthermore, scanning electron micrographs revealed membrane blebbing and apoptotic bodies on DOX-Ar-CC-NPs treated cells, signifying that DOX-Ar-CC-NPs induces apoptosis in breast cancer cells.


Our findings revealed the ability of DOX-Ar-CC-NPs to induce apoptosis in MCF-7 cells, which indicates the high potency of the Ar-CC-NPs understudy in drug delivery.

Keywords: Apoptosis, MCF-7 cell lines, Doxorubicin-Calcium Carbonate Nanoparticles, Drug delivery, Cytotoxicity