The Effectiveness of Gelatin Extract from Bone of Nile Tilapia Fish (Oreochromis niloticus) to accelerate Wound Healing 1 Student : Isnimyati Binti Joepri Supervisor : Associate Professor Ts. Dr. Reezal Ishak Introduction: Wound healing is a complex process in healthcare, and delay in wound healing continues to be a serious healthcare concern around the world. The Nile Tilapia fish (Oreochromis niloticus) has emerged as a potential alternative source for wound healing treatments, primarily due to its collagen-rich gelatin. Objective: This study aims to evaluate the effectiveness of gelatin extracted from Nile Tilapia fish bones in accelerating wound healing, offering an alternative to standard treatments. Method: The study employed an experimental design with twelve adult male Sprague Dawley rats, divided into three groups: a control group, a standard treatment group (Bepanthen-treated), and a group treated with gelatin extracted from Nile Tilapia fish bones. The process of gelatin extraction involved several steps, including treatment with sodium hydroxide, demineralization with EDTA, and lipid removal. The rat's wounds were assessed daily for nine days, and skin samples were collected for histological analysis. The study utilized statistical methods like ANOVA and t-tests to evaluate the treatment effects. Results: Results showed that on day 3, both bepanthen and gelatin treatments reduced wound size more than the control group. By day 9, the gelatin-treated wounds had the lowest closure percentage, indicating more efficient and complete healing compared to other groups. Histological evaluations supported these findings, with the gelatin-treated group showing higher average scores in wound healing, characterized by better re-epithelialization and keratinization. These results were statistically significant, highlighting the potential of gelatin treatment in wound healing. Conclusion: This study demonstrates the effective wound healing properties of gelatin extracted from Nile Tilapia fish bones when applied to inflicted wounds on the skin of the animal model. The results highlight its potential as an alternative treatment option. However, further investigation is necessary to validate its efficacy, safety, and understand its underlying mechanism of action. Keywords: Oreochromis niloticus, Gelatin, Bones, Wound healing, Reepithelialization YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 42
Acid-solubilized Collagen from Nile Tilapia Bones (Oreochromis nilocticus) Effects in Wound Treatment 1 Student : Muhammad Sabhi Nazmi Bin Mohd Sidek Supervisor : Associate Professor Ts. Dr. Reezal Ishak Introduction: In the medical field, wound healing is a challenging and important process. Collagen is the protein that is most prevalent in the human body. Cells like fibroblasts in the healing wound produce these collagens, which are then altered into complex morphologies. Objective: To promote new studies for more efficient wound treatment methods to have a much better understanding of the physiological processes that involved in normal wound healing using fish collagen. Method: Nine adult male Sprague Dawley rats were used in the experiment, and they were split into three groups: standard treatment (Bepanthen), collagen from Nile Tilapia fish bones, and a control group. The collagen extraction required a few steps, including lipid removal, demineralization with EDTA, and treatment with sodium hydroxide. For nine days, the rats wounds were examined every day, and skin samples were taken for histological examination. The study assessed the treatment effects using statistical techniques such as ANOVA and t-tests. Results: Although there was a tendency toward enhanced wound healing in the Bepanthen and collagen-treated group compared to the control and groups, statistical analysis using ANOVA did not reveal any significant differences between the groups at any time point. The Bepanthen group showed the lowest average wound size by day 9, followed closely by collagen suggesting possible healing improvements. These observations were supported by histological assessments, showing that both Bepanthen and collagen-treated group healed wounds more effectively on average, with better keratinization and re-epithelialization. Conclusion: In conclusion, despite the lack of statistically significant findings, the trends shown suggest that collagen from Nile Tilapia bones may have the ability to accelerate the wound healing. To confirm these initial findings and determine the beneficial effects of bone collagen from Nile Tilapia in wound healing applications, more research with bigger sample sizes and stricter control measures is needed. Keywords: Collagen, wound healing, keratinization, re-epithelialization YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 43
Effect of Black Seed (Nigella sativa) Infused Media on Cell Migration in Wound Healing Assay of Vero Cell 1 Student : Muhammad Syahirzi Bin Samsudin Supervisor : Associate Professor Ts. Dr. Reezal Ishak Introduction: Derived from Nigella sativa, a bioactive compound has shown promise in wound healing due to its antioxidant, anti-inflammatory, antibacterial, and immunomodulatory properties. This study delves into the therapeutic potential of N. sativa itself, examining its impact on Vero cell migration. Objective: This study aims to assess the wound healing potential of raw Nigella sativa-infused media on Vero cells. Specifically, the research investigates the effectiveness of a 24-hour infusion of N. sativa seeds in Dulbecco’s Modified Eagle Medium (DMEM) in promoting gap closure within scratched Vero cells, comparing its efficacy against a control group. The objective is to discern the specific influence of raw N. sativa on in vitro cell migration and its potential contribution to wound healing processes within a 24-hour timeframe. Method: N. sativa seeds underwent sterilization, were infused with DMEM for 24 hours, and then applied to scratched Vero cells in a 12-well plate. After 24 hours, gap closure was observed to evaluate the potential impact of raw N. sativa-infused media on Vero cell wound healing. Results: The 24-hour raw N. sativa-infused DMEM demonstrated superior gap closure compared to the control group, suggesting enhanced cell migration and potential wound healing benefits associated with the raw plant material. Conclusion: In summary, the 24-hour infusion of raw Nigella sativa into media exhibits promising wound healing potential by accelerating gap closure in scratched Vero cells. The inherent anti-inflammatory and antioxidant properties of N. sativa contribute to efficient wound closure, emphasizing the positive impact of raw N. sativa-infused media on cell migration. Further research is needed to optimize applications, shedding light on the potential of this natural compound to enhance wound-healing processes. Keywords: Nigella sativa, cell migration, wound healing, in-vitro, scratch assay, Vero cells YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 44
Exploring the Anxiolytic Potential: Evaluating the Effect of Panax ginseng and Ophiocordyceps sinensis Supplementation on Anxiety-Like Behavior in Animal Models Using Light Dark Box Test 1 Student : Izzah Hazwani Binti Moktar Supervisor : Mdm. Nabilah Hanim Mohd Sabri Introduction: This study explored the effect of supplementation rats with Panax ginseng (ginseng) and Ophiocordyceps sinensis (cordyceps) on anxiety-like behaviors. This study employed the light-dark box test to determine the potetial impact of these supplementation on anxiety-like behavior in rats. Objective: To determine the potetial impact of these supplementation on anxiety-like behavior in rats. Method: 18 rats were randomly divided into three groups: control, ginseng and cordyceps. After a 7-day supplementation period, anxiety-like behavior was evaluated by measuring the duration spent in the light compartments and the overall amount of entries from light to dark in the light-dark box. Results: The results revealed significant differences between the groups in anxiety-like behavior. The average of time spent in light compartment for control group is 21.63 seconds meanwhile it is 24.16 seconds and 24.46 seconds for ginseng and cordyceps respectively. The average of total entries from light to dark compartment were higher in ginseng and cordyceps (1.67 entries) compared to control (1.00 entries) indicating reduced anxiety. These findings suggest that both ginseng and cordyceps possess anxiolytic properties, potentially through their modulatory effects on the hypothalamic-pituitary-adrenal (HPA) axis and neurotransmitter systems. Conclusion: The findings of this study indicate a considerable reduction in anxiety-like behavior in rats supplemented with ginseng and cordyceps, as indicated by a large increase in time spent in the light compartment compared to the control group. Keywords: Panax ginseng, Ophiocordyceps sinensis, anxiety, light-dark box test YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 45
The Effects of Standardized Panax ginseng and Ophiocordyceps sinensis Supplementation on Prothrombin Time (PT) and Activated Partial Thromboplastin Time (APTT) in Rats 1 Student : Liyana Syakirah Binti Hashim Supervisor : Mdm. Nabilah Hanim Mohd Sabri Introduction: Conventional anticoagulant therapy presents safety issues since it increases the risk of bleeding. Ginseng and cordyceps, which are frequently used in traditional medicine for their potential health advantages, have been understudied in terms of their effect on blood coagulation. Objective: This study investigated on the effects of Panax ginseng and Ophiocordyceps sinensis supplementation on rats’ blood coagulation parameters, specifically prothrombin time (PT) and activated partial thromboplastin time (APTT). Method: In this study, rats were categorized into three groups: control, ginseng, and cordyceps, each consisting of six rats. Over a 7-day period, all rats received a 0.5 mL solution via a daily force-feeding procedure. The ginseng group received a solution containing 18 mg of ginseng, while the cordyceps group was given 36 mg. In contrast, the control group received a placebo solution. After the supplementing period, 3 mL of blood was drawn from each rat via heart puncture to collect plasma. Prothrombin time (PT) and activated partial thromboplastin time (APTT) were then measured using the Stago STart Max analyzer. All data are expressed as the mean±standard error (SE). Statistical analysis was performed using SPSS program (SPSS INC, ver. 27.0). A significance test (t-test) was set at a p-value of less than 0.05 (p<0.05). Results: The computed p-value for PT was 0.32, while for APTT it was 0.17, both over the significance level of 0.05. As a result, no statistically significant difference was found between the effects of ginseng and cordyceps on PT and APTT. Conclusion: These results suggest that both ginseng and cordyceps could not significantly extend blood clotting time in PT and APTT tests. Therefore, current results indicate that P. ginseng and O. sinensis supplements do not possess anticoagulation effects and do not have the potential to be an anticoagulation drug. Keywords: anticoagulant, thrombosis, Panax ginseng, Ophiocordyceps sinensis, prothrombin time (PT), activated partial thromboplastin time (APTT) YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 46
Acute Toxicity Effects of Panax ginseng and Cordyceps sinensis Supplementation on Hematological Parameters 1 Introduction: Aromatherapy has been defined, with some emphasizing its holistic approach, while others concentrate on its medicinal properties. Aromatherapy is also considered a complementary therapy or a form of alternative medicine. The ylang-ylang (Cananga odorata) flower is widely known for its captivating scent and is frequently used in aromatherapy and fragrance. Ylang-ylang aromatherapy has a distinctive aroma, full of natural substances, and has relaxing effects. Although it has great potential for aromatherapy, the cumulative effects of repeated inhalation of essential oils on lung tissues have not been adequately studied. Understanding how these organs respond to continuous exposure is crucial for evaluating overall safety. Objective: This study aimed to assess the histology of haematoxylin and eosin (H&E) stained rats’ lung sections following aromatherapy with Cananga odorata. Method: Ten adult male Sprague Dawley rats were randomly divided into the control (non-expose) and Cananga odorata aromatherapy-expose (COA) groups. Rats in the COA group were subjected to one-hour aromatherapy with Cananga odorata (50 μL) for fourteen days. On day 15, all rats were sacrificed, and lungs were collected and preserved in formalin for 48 hours. The well-fixed lungs were grossed and placed in tissue cassettes. Dehydration, xylene clearing, and paraffin infiltration were performed on the wellfixed lungs by using the Leica HistoCore tissue processor. Next, all lung specimens were blocked in paraffin wax, thinly sectioned (4-5 μm), and mounted on frosted glass slides. All lung sections were stained with H&E. Microscopic analysis was performed on all stained lung sections. Results: No histopathological changes were observed in all H&E-stained lung sections indicating that COA did not impose pulmonary toxicity. The H&E-stained lung sections of COA group exhibited normal alveolar sacs, alveoli and thin interalveolar septum lined by normal pneumocytes. Conclusion: Fourteen days of exposure to Cananga odorata aromatherapy did not induce pulmonary toxicity. Keywords: Aromatherapy, Cananga odorata, Ylang-ylang, Lung, Histology, Haematoxylin and Eosin (H&E) staining YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 47 Student : Nurul Fatin Binti Abdul Aziz Supervisor : Mdm. Nabilah Hanim Mohd Sabri
Effects of Panax ginseng and Ophiocordyceps sinesis Supplementation on Anxiety-Like Behaviour in Animal Models: A Study Using Elevated Plus Maze 1 Student : Nuradriyana Natasya Binti Mohd Rafid Introduction: Anxiety disorder is becoming more common and is affecting 4% of the global population, necessitating the exploration of novel therapeutic interventions. This thesis studies the potential anxiolytic effects of Panax ginseng and Ophiocordyceps sinensis supplementation in animal models, evaluated using the elevated plus maze. Panax ginseng and Ophiocordyceps sinensis, are both traditional medicinal herbs that are well known for their adaptogenic and neuroprotective properties, emerge as promising candidates for anxiety management. Method: In this study, 18 males Sprague-Dawley rats were categorised into three groups: control, ginseng, and cordyceps, with each consisting of six rats. Behavioural observations were performed using elevated plus maze before supplementation with treatment. Over a 7-day period, all rats received a 0.5 mL solution via a daily force-feeding procedure. The ginseng group received a solution containing 18 mg of ginseng, while the cordyceps group was given 36 mg. Post supplementation assessments using the Elevated Plus Maze test were performed again. The behavioural outcomes were evaluated by measuring the number of entrances made by the rats into each arm of the maze, including both closed and open arms. Results: During post-treatment behavioural assessments, rats showed a significant decrease in anxiety-like behaviour after receiving Panax ginseng and Cordyceps sinensis supplements. This finding indicates the efficacy of Panax ginseng and Cordyceps sinensis in reducing anxiety-related behaviours in the experimental subjects. Conclusion: This research could contribute to the formulation of natural supplements for anxiety management, providing valuable insights for both traditional medicine and modern pharmacology. Furthermore, the study may pave the way for future clinical trials, fostering a more comprehensive understanding of the therapeutic potential of these herbal supplements in addressing anxiety-related conditions. Keywords: Panax ginseng, Ophiocordyceps sinensis, Anxiolytic Effects YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 48 Supervisor : Mdm. Nabilah Hanim Mohd Sabri
Aronia berries Extract as Nutrition Remedy for Urinary Tract Infection 1 Student : Marini Nurshuhadah Binti Abdullah Introduction: Aronia berries, or chokeberries, are small, dark fruits that grow on shrubs of the Rosaceae family and are native to North America. Aronia berries can act as a remedy for urinary tract infection (UTI) due to the high quinic acid, preventing the bacteria that cause bladder issues. UTI can affect both the upper and lower urinary tract, which includes the kidneys and bladder infection. In 2019, more than 404.6 million individuals had UTIs globally, and nearly 236,786 people died of UTIs, contributing to 5.2 million DALYs. The most common treatment for a UTI involves a course of antibiotics. Long-term antibiotic therapy is an effective way to treat recurrent UTI, but it has drawbacks, where it can develop antibioticresistance. The potential of Aronia berries, specifically as a nutrition remedy, needs to be explored further. Objective: The purpose of this study is to determine the effectiveness of Aronia berries extract as a nutritional remedy for urinary tract infections. In this study, the UTI bacteria used were Escherichia coli, Staphylococcus aureus, and Proteus mirabilis. Method: The antibacterial activity was tested using the well diffusion method, Minimum Inhibition Concentration (MIC) and Minimum Bactericidal Concentration (MBC). As for the well diffusion process, for each bacterium, there is a zone of inhibition recorded surrounding the impregnated disk with the Aronia berries extract. The Minimum Inhibitory Concentration (MIC) of the extract was assessed, yielding notable findings for bacterial strains: E. coli (p=0.002), S. aureus (p=0.008), and Proteus mirabilis (p=0.003). Results: The MIC results revealed significant differences in susceptibility among bacterial strains, with E. coli, S. aureus, and Proteus mirabilis exhibiting distinct responses (p<0.05). Consequently, the MBC results for each bacteria indicate no growth of bacteria at concentrations of 50%, 25%, 12.5% and 6.25%, demonstrating the efficacy of the tested substance in inhibiting bacterial growth across various concentrations. Conclusion: In conclusion, is Aronia berries extract effective as a nutritional remedy for urinary tract infection. Keywords: Aronia berries, urinary tract infections, antibiotic resistance, Escherichia coli, Staphylococcus aureus, and Proteus mirabilis YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 49 Supervisor : Mr. Hanan Kumar S/O Gopalan
Wound Healing Assessment in Rats Treated with Topical Metformin 1 Student : Muhammad Aiman Hafizi Bin Saiful Azahar Introduction: Wound healing is an intrinsic physiological process in the human body that arises in response to tissue injury. Metformin's therapeutic repertoire extends beyond glycemic control, encompassing diverse antiinflammatory and immunomodulatory properties. Its action is multifaceted, targeting both cellular metabolic pathways and immune cell function. Metformin has recently garnered significant interest for its potential wound- healing properties demonstrated in preclinical studies. Objective: This study aims to address these critical research gaps by conducting a comprehensive investigation into the efficacy and potential use of topical metformin in the context of wound healing. Method: In this study, a total of 12 Sprague-Dawley rats weighing from 200 to 300g and free from any abnormalities were used to create the wound excisional model. A wound excision was performed using an 8 mm diameter, 2 mm depth medical-grade biopsy punch on the dorsum of the rats. Daily topical applications of metformin onto the wound bed were conducted for a duration of 21 days. At the end of the 21-day period, rats were euthanized using an overdose of ketamine, and the wounded area was biopsied for subsequent histological studies. Wound closure measurements and the healing duration data were gathered for descriptive statistical analysis. Results: The investigation revealed that the topical administration of metformin induced a significant improvement in wound healing, accompanied by notable alterations in ultrastructural remodeling post-treatment. Histopathological analysis provided compelling evidence supporting the ability of topical metformin to stimulate collagenogenesis, thereby promoting tissue regeneration. Conclusion: This study concludes that the topical administration of metformin stands as an exceptionally valuable therapeutic strategy for facilitating efficacious wound healing and robust tissue regeneration. Keywords: Wound healing, Skin, Metformin YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 50 Supervisor : Mr. Hanan Kumar S/O Gopalan Co-Supervisor : Dr. Faiza Che Harun , Associate Professor Dr. Yanti Rosli, Dr. Pang Jyh Chyang
Morphological Evaluation of Topical Metformin Efficacy in Treating Atopic Dermatitis in Rats 1 Student : Siti Nurfarrisha Shahira Binti Thajudeen Objectives: This study aims to assess the efficacy of topically administered metformin preparation in the treatment of atopic dermatitis (AD) induced in rats using 1-chloro-2,4-dinitrobenzene (DNCB). Morphological evaluations were systematically documented over a 21-day treatment period to monitor the progression of healing. Method: Rats used in this study were screened for anomalies and subsequently sensitized by the application of 1% DNCB for a minimum of 4 weeks. Upon visual confirmation of AD marked by the manifestation of scaly and keratinous skin eruptions, rats were subjected to the topical application of a metformin preparation. The healing progress facilitated by metformin application was photographically documented at intervals of 1, 7, 14, and 21 days, respectively. Skin biopsies were collected after the 21-day treatment period, followed by processing and staining with H&E and Toluidine Blue for subsequent histological analysis. Results: The findings from the study demonstrated a significant reduction in skin inflammation and an overall enhancement in the healing of skin architecture following the application of metformin. Histological analysis utilizing H&E and Toluidine blue staining revealed pronounced reparative effects associated with metformin application, mitigating the detrimental effects induced by atopic dermatitis (AD). The significant reduction in inflammation and the comprehensive improvement in the healing of skin architecture observed provide substantial support for the potential efficacy of topically applied metformin. Conclusion: These findings yield valuable insights into the reparative effects of metformin at the cellular level, consequently advocating for its exploration in the advancement of dermatological treatment strategies. Keywords: Atopic dermatitis, 1-chloro-2,4-dinitrobenzene, metformin, topical treatment, morphology, inflammation, histopathological analysis YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 51 Supervisor : Mr. Hanan Kumar S/O Gopalan Co-Supervisor : Associate Professor Dr. Yanti Rosli
Assessment of Burn Wound Healing in Rats Treated with Topical Metformin 1 Student : Nurul Izzah Binti Mohd Zaki Introduction: Burn injuries pose a substantial worldwide health concern, contributing to considerable morbidity and mortality. These injuries frequently lead to delayed wound healing, with an elevated risk of bacterial infections. Metformin, a primary medication prescribed as a first-line treatment for type 2 diabetes (T2DM), has garnered extensive interest due to its potential anti-inflammatory properties, bacteriostatic effects, and promotion of cell proliferation. Objectives: The objective of the current study is to assess the therapeutic impact of topically applied metformin on second-degree burn wounds using a rat model.. Method: The study involved 12 male Sprague Dawley rats devoid of anomalies. Superficial second-degree burns, measuring 2.4 cm, were induced by applying a heated coin to the dorsal region of the rats backs. Following burn infliction, the affected areas were treated with topically applied metformin for a duration of 21 days. Upon completion of the treatment period, therats were euthanized, and skin samples were collected for subsequent histological examination. Results: Microscopic assessment revealed remarkable and noticeable changes in tissue regeneration, collagen deposition, and a noteworthy improvement in ultrastructural reparative conditions. Statistical analysis demonstrated a significant reduction in wound size, indicating a positive impact of the topical metformin treatment on the healing process (P<0.05). Conclusion: This study underscores the efficacy of topical metformin preparation as a viable therapeutic option for the treatment of burn wounds. The innovative nature of this treatment presents a promising avenue for alleviating the suffering of burn victims, thereby establishing it as a valuable and advantageous alternative in the realm of burn wound management. Keywords: metformin, burn, wound healing, rat YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 52 Supervisor : Mr. Hanan Kumar S/O Gopalan Co-Supervisor : Associate Professor Dr. Yanti Rosli, Dr. Pang Jyh Chyang, Dr. Faiza Che Harun
Wound Healing Assessment in Diabetic Rats Treated with Topical Metformin 1 Student : Syahidatun Nadhirah Binti Ahmad Amir Introduction: Diabetes constitutes a metabolic anomaly characterized by dysregulated glucose metabolism. Type 2 Diabetes Mellitus (T2DM) exemplifies a protracted metabolic aberration hallmarked by heightened blood glucose levels, denoting hyperglycemia, attributable to compromised insulin utilization. Within the diabetic milieu, individuals routinely contend with impaired wound healing, a phenomenon intricately linked to diverse diabetes-associated factors. Understanding the nuanced interplay of these elements is imperative for elucidating the impediments to the wound healing process in diabetes. Objectives: This investigation seeks to ascertain the potential efficacy of metformin application in fostering wound healing within the context of diabetes mellitus (DM) in rats. Method: In this investigation, 12 healthy male SpragueDawley rats, weighing between 250-350 grams and free of any anomalies, served as experimental subjects. Following a one-week acclimatization period, the rats underwent weighing and subsequent categorization. Diabetes was induced through a single intraperitoneal (IP) injection of 65 mg/kg Streptozotocin (STZ). Fasting blood glucose levels were measured four days’ post-injection, and rats with levels exceeding 15 mmol/L were included in the study. Wound excision was performed using an 8 mm diameter, 2 mm depth medical-grade biopsy punch on the dorsal torso of rats. Daily topical application of metformin onto wound bed were carried out for 21 days. At the end of Day 21 euthanasia was performed by an overdose of Ketamine and biopsy specimens were obtained for subsequent histological staining and analysis. Result and Conclusion: The present study yields compelling evidence indicating that the topical application of metformin exerts a notable influence on the wound healing dynamics in diabetic rats. Observable pathological alterations include a statistically significant reduction in wound diameter is indicative of an accelerated healing process. Additionally, microscopic examination of tissue specimen’s reveals marked improvements in histological architecture,underscoring the beneficial impact of metformin on the reparative mechanisms within the diabetic wound microenvironment. Keywords: Type 2 Diabetic Mellitus (T2DM), Wound Healing, Metformin YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 53 Supervisor : Mr. Hanan Kumar Gopalan Co-Supervisor : Associate Professor Dr. Yanti Rosli, Dr. Faiza Che Harun, Dr. Pang Jyh Chyang.
Phytochemical Analysis of Citrus hystrix DC. Essential Oil and Modulation of Cognitive Performance by Aromatherapy in Rats 1 Student : Muhammad Danial Haziq Bin Zulkifli Introduction: Aromatherapy is a non-pharmacological approach that utilizes essential oils (EO) to enhance the quality of life. Studies have claimed that the essential oil of Citrus hystrix DC. peel, or kaffir lime peel contains bioactive compounds with neuroprotective properties. Aromatherapy with Citrus hystrix DC has not been properly tested on cognitive performance. Objective: This study aims to analyze the phytochemical constituents of Citrus hystrix DC. peel essential oil (CHEO) and investigate the modulation of cognitive performance by aromatherapy of CHEO in rats. Method: Hydrodistillation and gas chromatography-mass spectrometry were used to extract and analyze CHEO. Rats were exposed to one hour of CHEO aromatherapy (50 μL) daily for two weeks. The novel object recognition test (NORT) was performed, and the discrimination index (DI) was calculated to evaluate recognition memory in rats. Brain-derived neurotrophic factor (BDNF), a biomarker of neuroplasticity and brain health, was measured using enzyme-linked immunosorbent assay (ELISA). Histological analysis of neuronal structure in the hippocampal region was performed using hematoxylin and eosin staining. Results: GC-MS analysis identified 20 volatile chemicals, with β-pinene (32.96%), limonene (24.34%), and sabinene (21.14%) being the most abundant. The exposed group (56.3 ± 35.3; p<0.005) outperformed the control group (-13.7 ± 39.7) in the NORT with a significantly higher DI. A positive DI suggests novel object selection and intact recognition memory. Additionally, the CHEO-exposed rats had higher BDNF levels (685.4 ± 57.5 pg/mg protein; p<0.005) than control rats (586.6 ± 21.3 pg/mg protein). Histological examination of neurons in both the control and CHEO-exposed groups revealed healthy neurons in the hippocampal regions. Conclusion : In summary, the study indicated that the CHEO mainly consists of β-pinene, limonene, and sabinene. This study also proved that the aromatherapy of CHEO positively modulates cognitive performance and has no detrimental effect on the hippocampus.This essential oil’s potential to cure neurologicalillnesses, particularly Alzheimers dementia,should be further studied. Keywords: Citrus hystrix DC. Peel Essential Oil (CHEO), Neuroprotective, BrainDerived Neurotrophic Factor (BDNF), Novel Object Recognition Test (NORT) YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 54 Supervisor : Dr. Teh Rasyidah Ismail Co-Supervisor : Dr. Dwiarso Rubiyanto
Cananga odorata Essential Oil: Unravelling Phytochemical Components and Its Cognitive Effects in Rats 1 Student : Muhammad Faqhrul Fahmy Bin Arbain Introduction: Cognitive health has emerged as prominent concern in the field of global public health. The essential oil of Cananga odorata (EO-CO), derived from fragrant blossoms of the ylang-ylang tree and containing variety of bioactive compounds, gaining recognition for its potential impact on memory and cognition. Despite its potential neuroprotective properties, minimal research has been conducted into the efficacy of EO-CO in sustaining neuronal health. Objective: This research aimed to identify the phytochemical components of EO-CO and investigate the cognitive effects of EO-CO in Sprague-Dawley rats. Method: The EO-CO was extracted using hydrodistillation, and its components were identified using gas chromatography-mass spectrometry (GC-MS). For 14 days, rats were given EO-CO (50 μL) by inhalation (1 hour each day). The rats' recognition memory was evaluated by conducting the novel object recognition test (NORT) and determining the discrimination index. A positive DI score indicates that the recognition memory is intact. The enzyme-linked immunosorbent assay (ELISA) evaluated BDNF levels in the hippocampus. Histological study of neuronal structure in hippocampal areas was also performed using hematoxylin and eosin staining. Results: The GC-MS analysis detected 25 phytochemical components, with linalool (38.37%), trans caryophyllene (23.82%), and germacrene D (13.69%) being the most prevalent. The EO-CO exposed group had significantly higher discriminating index (80.0 ± 56.73; p<0.005) than the control group (-13.7 ± 39.7). The EO-CO exposed group exhibited significantly higher levels of BDNF (635.4 ± 27.1 pg/mg protein; p<0.005) than the control group (586.6 ± 21.3 pg/mg protein). Histological investigation revealed no significant neuronal damage in the EO-COexposed group, implying that EO-CO did not produce neurotoxicity. Conclusion : Therefore, this study discovered that essential oil extracted from Cananga odorata flowers has neuroprotective properties, signifying potential improvements in cognitive function. Future research should focus on the essential oil's possible therapeutic effects in neurological conditions such as Alzheimer's disease. Keywords: Cananga odorata, Essential Oil, Brain-Derived Neurotrophic Factor, Novel Object Recognition Test, Gas Chromatography-Mass Spectrometry, Neuroprotective YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 55 Supervisor : Dr. Teh Rasyidah Ismail Co-Supervisor : Dr. Dwiarso Rubiyanto
Characterization of Bioactive Constituents in Ylang-ylang (Cananga odorata) Essential Oil and Their Potential Anxiolytic Effects in Rats 1 Student : Muhammad Nazrel Bin Zulkifli Introduction: This study focuses on the exploration of potential anxiolytic effects stemming from the inhalation of Ylang-ylang essential oil (YEO) in rats. Extracted from the fresh and ripe blossoms of Cananga odorata, YEO is widely employed in the fragrance industry. As awareness grows regarding the efficacy of plantderived essential oils in addressing mood disorders and enhancing sleep, aromatherapy stands out as a promising non-pharmacological approach for managing anxiety. Objective: This research aims to investigate the anxiolytic properties of YEO when administered through aromatherapy. Method: The meticulous extraction of YEO involved a hydro-distillation process, followed by a detailed analysis using gas chromatography–mass spectrometry (GC–MS). Over a span of fourteen days, rats were exposed to one hour of YEO (50 μL) aromatherapy in an inhalation chamber. The anxiolytic impact of YEO was then assessed through the light-dark box (LDB) test. Post-experiment, rat brains were collected for serotonin quantification in the cerebral cortex via the ELISA method. Additionally, histological analysis was conducted on the collected brains. Statistical analysis, utilizing an independent samples t-test, was employed to discern significant differences. Results: GC-MS analysis revealed that Linalool constituted the highest bioactive composition of YEO at 38.37%. The LDB test demonstrated that rats exposed to YEO spent significantly more timein light areas (38.90 ± 9.21%), indicating lower anxiety compared to control rats (8.84 ± 3.97%). Moreover, serotonin levels in YEO-exposed rats were significantly higher (2788 ± 224 ng/mg protein) than in control rats (1910 ± 184 ng/mg protein). Histological observations showed no discernible difference from the control group, suggesting the absence of neurotoxic effects induced by Ylang-ylang essential oil. Conclusion: This study revealed that the main bioactive composition of Ylangylang (Cananga odorata) essential oil was Linalool. This study also proved that the Ylang-ylang (Cananga odorata) essential oil has potential anxiolytic effects. Keywords: Ylang-ylang, Cananga odorata, Essential oil, Aromatherapy, Linalool, Mood Disorders YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 56 Supervisor : Dr. Teh Rasyidah Ismail Co-Supervisor : Dr. Dwiarso Rubiyanto
Chemical Composition and Anxiolytic-like effect of Citrus hystrix DC. Peel Essential Oil in Rats 1 Student : Sharifah Suhana Syed Abd Rahman Introduction: Citrus hystrix DC., known as kaffir lime, is a prominent citrus species in Southeast Asia. Its fruit and leaves play a significant role in Asian cuisine, and its peel can be distilled to extract essential oil. Despite centuries of traditional use in treating illnesses, there is a research gap in understanding its anxiolytic-like effects. Current research emphasizes antibacterial properties primarily, neglecting the comprehensive exploration of its anxiolytic potential. Objective: This study aims to identify key bioactive compounds of Citrus hystrix DC. peel and investigate its anxiolytic-like effect in rats. Method: The Citrus hystrix DC. peel essential oil (CEO) was obtained by hydrodistillation and analyzed by Gas Chromatography/Mass Spectrometry (GC-MS) to identify its volatiles. Ten adult male Sprague Dawley rats were randomly assigned into two groups: Control and CEO-exposed. The CEO-exposed rats were subjected to 1 hour/day CEO (50 μL) inhalation. After 14 days of inhalation, the light-dark box (LDB) test was conducted to evaluate the anxiolytic-like effect of CEO in rats. Enzyme-linked immunosorbent assay (ELISA) was performed to measure the serotonin levels in the cerebral cortex homogenate. The neuronal structure in the prefrontal cortex was visualized using hematoxylin and eosin (H&E) staining. Results: GC-MS analysis identified 20 compounds with CEO predominated by βpinene (32.96%), limonene (24.34%), sabinene (21.14%), and citronellal (13.31%). The LDB test showed CEO-exposed group (41.14 ± 17.34 %, p<0.05) had a higher percentage of light-to-dark ratio than the control group (8.84 ± 3.97 %). The serotonin level was significantly higher in the CEO-exposed group (2563 ± 215 ng/mg protein, p<0.05) than in the control group (1910 ± 184 ng/mg protein). Additionally, H&E-stained sections of the prefrontal cortex revealed no histopathological changes in both groups. Conclusion: The Citrus hystrix DC. peel essential oil is predominantly composed of β-pinene, limonene, sabinene, and citronellal. It exhibits anxiolytic-like properties with no detrimental effects on neuronal cells in the prefrontal cortex. Keywords: Hydrodistillation, essential oil, Citrus hystrix DC., anxiolytic-like effect, serotonin, light-dark box test YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 57 Supervisor : Dr. Teh Rasyidah Ismail Co-Supervisor : Dr. Dwiarso Rubiyanto
Psychoactive Potential: Evaluating The Anxiolytic Properties of Cananga odorata and Citrus hystrix DC. Essential Oil Blends in Rats 1 Student : Siti Fariha Binti Ghazali Introduction: Cananga odorata and Citrus hystrix DC. are native to tropical Asia and are believed to alleviate depression and anxiety symptoms. However, there is a research gap on the synergistic effects of blended essential oils. Evaluating the anxiolytic effect in rats is crucial for developing effective therapeutic strategies. Objective: This study aims to investigate the possible anxiolytic effect of essential oil blends of Cananga odorata flowers and Citrus hystrix DC. peel (EOB) in rats. Method: Both EOs were extracted by hydrodistillation technique and were subjected to Gas Chromatography/Mass Spectrometry (GC/MS) for identification of their volatile compounds. Sprague Dawley rats (n=10, ♂ ) were randomly divided into control and EOB-exposed groups. Rats in the EOB-exposed group were subjected to 1 hour/day EOB (50 μL: 50 μL) inhalation for fourteen days. The light-dark box (LDB) test was conducted to evaluate the anxiolytic-like effect of EOB. The study utilized ELISA to measure serotonin levels in cerebral cortex homogenate, while H&E staining was used to visualize the morphology of prefrontal cortex neurons. Results: The GC/MS evaluation reported the main volatile of Cananga odorata EO was Linalool (38.37%) and the main volatile of Citrus hystrix DC. EO was Beta Pinene (32.96%). The LDB test revealed that EOB-exposed rats had a higher percentage of light-to-dark ratio (37.1 ± 16.1%; p<0.05) than control rats (8.8 ± 4.0%; p < 0.05). The study found that rats exposed to EOB spent more time in the light area, indicating a decrease in anxiety levels. The EOB- exposed rats had higher serotonin levels (3227 ± 457 ng/mg protein; p<0.05) than control rats (1910 ± 184 ng/mg protein). The study found no histopathological changes in prefrontal cortex sections in both groups, indicating that EOB did not cause neurotoxicity. Conclusion: The study reveals that Cananga odorata and Citrus hystrix DC. essential oil blends have anxiolytic properties, suggesting a potential mood-enhancing method. Keywords: Cananga odorata, Citrus Hystrix DC., Anxiolytic, Essential Oil Blends, Linalool, Beta Pinene YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 58 Supervisor : Dr. Teh Rasyidah Ismail Co-Supervisor : Dr. Dwiarso Rubiyanto
Detecting The Expression Of C-Jun In Glucose-Induced Schwann Cells 1 Student : Muhammad Redwan bin Safie Supervisor : Dr. Jalilah binti Idris Co-Supervisor : Dr. Muhammad Fauzi bin Daud Background: Diabetic peripheral neuropathy (DPN), a complication of diabetes mellitus has been proven to cause denervation of the Schwann cells. Schwann cells will stimulate myelin and non-myelin (Remak) that will change into a specialised cell phenotype that supports damage healing. Axon regeneration, the survival of injured neurons, and the reinnervation of the target distal to the lesion are all made possible by the signals and location cues supplied by the repair Schwann cells. Schwann cells will undergo de-differentiation to repair itself, with the help of transcription factor c-Jun. Currently, there are lack of studies to understand about the role of c-Jun in nerve injury caused by high glucose concentration. Objective: This project aims to investigate the co-relation between level of expression of c-Jun in the glucose level in the Schwann cells. Method: Primers targeting c-Jun gene were designed by using the Primer-BLAST software. Cell pellets from Schwann cells cultured in different concentration of glucose were lysed, followed by RNA extraction and cDNA synthesis. The expression of c-Jun was to be monitored using the quantitative polymerase chain reaction (PCR), and differential CT value was calculated. One-way ANOVA from GraphPad Prism was used for statistical analysis Results: The RNA yield for Schwann cells cultured in 25 mM (control) and 50 mM were less than 9 ng, which results in low value for A260/230. qPCR result show that the expression of c-Jun was elevated by two-fold at high concentration of glucose (50 mM), compared to the control (25 mM) (n=2). The high expression of c-Jun gene suggests that Schwann cells de-differentiate due to the damage inflicted from the DPN. Conclusion: The c-Jun gene expression appears to increase at higher glucose concentration, however, more study needs to be repeated in order to conduct statistical analysis. Keywords: Schwann cells; De-differentiation; c-Jun; qPCR YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 59
Optimization on the PCR Conditions for The Amplification of eGFP 1 Student : Muna Zulaikha Binti Mohd Zamri Supervisor : Dr. Jalilah binti Idris Introduction: Enhanced green fluorescent protein (eGFP) is a genetically modified version of the green fluorescent protein (GFP) that was originally found in A. victoria jellyfish. It is frequently employed as a reporter gene in the field of cloning. eGFP enables researchers to observe the subcellular distribution of the protein of interest without the need for immunohistochemical staining. Objective: Thus, as this project serves as a preliminary investigation, its objective is to determine the most optimal setting for the amplification of the eGFP human gene using the polymerase chain reaction (PCR). Method: Firstly, primers specific to the human eGFP gene were identified using the NCBI Primer-BLAST software. The eGFP sample was obtained from the pUC57 plasmid. EcoRI restriction enzyme was used to cut 1 ug of the DNA plasmid. The PCR reaction was performed using 1 ug of DNA, using the following parameters: 94°C for 30 seconds, 94°C for 30 seconds, 60°C for 1 minute, 68°C for 1 minute, 68°C for 5 minutes, followed by gel electrophoresis. Gel Doc was utilised to detect amplicons on the gel. Results: The outcome of the digestion process produced a DNA concentration of 95.9 ng/ul, indicating a somewhat low level of purity. A single prominent band measuring 720 base pairs was detected on the gel, indicating that the primers used may possess a high degree of specificity for the eGFP gene. Additionally, two other unidentified bands were observed. Conclusion: In conclusion, the ideal PCR conditions were effectively determined. However, alternative primers could be employed as a result of the presence of primer dimer. Keywords: eGFP, optimization, PCR, cloning YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 60
Detecting the Expression of Krox20 gene in GlucoseInduced Schwann Cells 1 Student : Nooratiqah Binti Mohamad Azmi Supervisor : Dr. Jalilah binti Idris Co-Supervisor : Dr. Muhammad Fauzi Daud Introduction: Diabetic peripheral neuropathy (DPN) is the most common compilation of diabetes that caused by nerve injury from high blood glucose levels, primarily affecting the legs, feet, hands and arms. DPN is distinguished by the absence of myelination on neurons, resulting in decreased nerve conduction velocity, and axonal atrophy. Schwann cells function as the myelinating cell of the peripheral nervous system and contribute to the development of DPN. Schwann cells rely on the presence of a specific protein known as Krox20, which acts as a transcription factor for controlling cell death and proliferation, promoting myelin formation, and determining the level of maturity of Schwann cells. Objective: DPN induces downregulation of Krox20 expression, leading to the suppression of myelin production, increased Schwann cell proliferation, and death. Method: Primer targeting Krox20 was designed using the Primer-BLAST software and purchased from IDT company. Next, RNA was extracted from glucose-induced Schwann cell culture (25 mM [control] and 50 mM of glucose), and the concentration and purity were assessed using nanophotometer. Then, cDNA was synthesized and used in quantitative polymerase chain reaction (qPCR). Results: Low number of RNA was extracted from cultured Schwann cells. The qPCR analysis indicates that the expression of the Krox20 gene is reduced in high glucose conditions (n=2). This suggests that elevated glucose levels (50 mM) cause damage to Schwann cells, leading to a decrease in the expression of the Krox20 gene, which in turn promotes the occurrence of demyelination and nerve repair. Conclusion: The expression of Krox20 seems to be diminished when exposed to high levels of glucose. However, it is necessary to duplicate this study in order to do a statistical analysis. Keywords: Diabetic peripheral neuropathy, Schwann cells, Krox20, glucoseinduced Schwann cell culture, qPCR YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 61
Determining The Expression Of CNOT3 Gene in an InVitro Spinal Cord Injury Model 1 Student : Nor Jannatul Husna Binti Jamaludin Supervisor : Dr. Jalilah binti Idris Introduction: More than 350,000 people worldwide are permanently disabled as a result of spinal cord injury (SCI). SCI occurs when the spinal cord is injured, thereby triggering cell death. The severity of the SCI increases with the extent of cell death. Thus, therapeutic intervention is critical for stopping the loss of spinal cord cells. The necroptotic cell death pathway was strongly activated in the cell model of SCI. CNOT3 is one of few RNA-binding protein that regulates necroptosis. Objective: Therefore, this study aimed to assess the expression of the CNOT3 gene in a cellular model of SCI using quantitative polymerase chain reaction (qPCR). Method: Primers against mouse CNOT3 gene were designed using the National Centre for Biotechnology Information (NCBI) website. Subsequently, RNA samples from two conditions were subjected to conversion into cDNA transcripts using the LunaScript kit. The conditions included: i) injured differentiated NSC34 cells; and ii) differentiated NSC34 cells without injury (negative control). qPCR procedure was carried out utilising Luna Universal qPCR Master Mix. The differential CT value was calculated followed by statistical analysis via one-way ANOVA using GraphPad Prism. Results: The expression level of the CNOT3 gene remains unaltered in the cellular model of spinal cord injury (SCI), despite significant activation of necroptosis. This indicates that CNOT3 does not have a role in controlling necroptosis in the SCI. Conclusion: Our findings indicate that CNOT3 does not have a role in controlling cell survival in the SCI model. The fact that it is not involved in regulating cell survival in this particular injury model indicates the necessity to investigate other molecular targets. Keywords: CNOT3, cell death, necroptosis, spinal cord injury, quantitative polymerase chain reaction (qPCR) YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 62
Determining The Expression of Smn1 Gene in an In-Vitro Spinal Cord Injury Model 1 Student : Norain Raihana Risma Binti Mohd Salleh Supervisor : Dr. Jalilah binti Idris Introduction: Spinal cord injuries (SCIs) is trauma to the spinal cord which might lead to motor impairment such as paralysis. SCI inflict cellular damage such as ischemia, excitotoxicity, and extensive cell death. Survival motor neuron (SMN) is highly expressed is spinal cord, and is associated with regulating cell survival. Preliminary study shows that Smn1 gene, coding for SMN protein, was elevated in animal SCI model. Objective: Therefore, this study aimed to determine the expression of Smn1 gene in an in-vitro SCI model. Method: Primers targeting for Smn1 gene were identified from NCBI Primer-Blast software. Next, RNA samples (control, 1, 0.5 and 1 uM) from kainic acid-treated mouse motor neuron-like hybrid cell line, NSC34 cells, were reverse transcribed into cDNA. The expression of Smn1 was measured against beta-actin using quantitative polymerase chain reaction (qPCR). Results: The expression of Smn1 was significantly elevated by two-fold in 0.1 mM kainic acid. This suggests that SMN, encoded by Smn1, play a role in SCI. Conclusion: This research contributes to our understanding of the molecular landscape of spinal cord injuries, indicating that SMN1 may be a key player in the cellular response to such traumatic events. Unraveling the intricacies of SMN1 expression post-SCI moves us closer to developing innovative strategies to modulate this protein, potentially improving outcomes and quality of life for individuals affected by spinal cord injuries. Keywords: Spinal cord injuries, Survival Motor Neuron 1 (SMN1), In-vitro model, Quantitative polymerase chain reaction (qPCR), Neurological damage YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 63
In Vitro Study Of Anticoagulant and Thrombolytic Activity of Nephelium lappaceum Peel Extract on Human Blood 1 Student : Nur Atira Binti Azmie Supervisor : Dr. Norhaida Binti Che Azmi Introduction: In Malaysia, venous thromboembolism (VTE) is the main factor contributing to maternal mortality. Conventional anticoagulants such as low molecular weight heparin, edoxaban, and rivaroxaban that are used to treat VTE can cause unwanted side effects. Plants can be an alternative for the new natural anticoagulant. The peel of Nephelium lappaceum (Rambutan) possesses antibacterial, antioxidant, antidiabetic, and anticancer activities. Objective: This study aims to investigate the anticoagulant and thrombolytic activity of Nephelium lappaceum peel extract, as previous research has demonstrated that plants with antioxidant properties also possess anticoagulant effects. Method: The anticoagulant activity of Nephelium lappaceum peel extract is evaluated by measuring the prothrombin time (PT) and activated partial thromboplastin time (aPTT) at three different concentrations (5 mg/ml, 10 mg/ml, and 20 mg/ml). Meanwhile, thrombolytic activity of Nephelium lappaceum peel extract is observed through clot lysis at concentration of 20 mg/ml. Results: In PT test, all three concentrations of Nephelium Lappaceum peel extract (5 mg/ml, 10 mg/ml, and 20 mg/ml) exhibit no coagulation even after passing the end point, which is 20 minutes. This show that they significantly prolonged the PT (p<0.05). No coagulation within the end point is recorded in aPTT test at concentrations of 10 and 20 mg/ml. However,clotting can be observed at concentration of 5 mg/ml (18.37 ± 0.30) which is significantly different compared to the negative control (0.40 ± 0.02). All three concentrations of Nephelium lappaceum peel extract (5, 10, and 20 mg/ml) can prolong aPTT significantly (p<0.05). In thrombolytic assay, Nephelium lappaceum peel extract at 20 mg/ml has a poor mean of clot lysis with just 32.51% compared to the positive control's, 36.37%. This indicates that it does not have significant thrombolytic activity (p= 0.793). Conclusion: Therefore, we can conclude that Nephelium lappaceum peel extract exhibit strong anticoagulant activity, but lack of thrombolytic property. Keywords: Activated partial thromboplastin time (aPTT), Anticoagulant activity, Methanolic extract, Nephelium lappaceum (rambutan), Prothrombin time (PT), Thrombolytic activity, Venous thromboembolism (VTE) YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 64
In Vitro Study of Anticoagulant and Thrombolytic Activity of Citrus hystrix Leaf Extract on Human Blood 1 Student : Nur Atira Binti Riza Effendi Supervisor : Dr. Norhaida Binti Che Azmi Introduction: The normal hemostasis indicates that the body has a balance natural occurring procoagulant and anticoagulant mechanisms. The shifting or interference of these balance naturally occurring mechanisms leads to increasing risk of bleeding or thrombosis. Citrus hystrix which also known as “kaffir lime” have the properties that can prevent from the occurrence of thrombosis. Objective: To investigate the anticoagulation and thrombolytic activities of Citrus hystrix leaf methanolic extract through in vitro study. Method: Citrus hystrix leaf methanolic extract was produced. Blood samples were withdrawn from ten healthy adult human volunteers. The anticoagulation activity of C. hystrix methanolic extract was identified through PT and aPTT test with concentration of 5, 10 and 20 mg/mL while thrombolytic activity was tested through clot lysis assay with concentration of 20 mg/mL. Results: Citrus hystrix leaf methanolic extract have a significant (p<0.05) anticoagulant activity with increasing prolongation clotting time at concentration of 20 mg/mL compared to control groups. However, it also recorded that this methanolic extract have insignificant (p>0.05) thrombolytic activity compared to the control groups. Conclusion: From this research, kaffir lime leaf extract proved to have a remarkable anticoagulant properties and yet its thrombolytic activity needed to be further studied. Keywords: Citrus hystrix, anticoagulant, thrombolytic, PT, aPTT, p value YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 65
In Vitro Study of Anticoagulant and Thrombolytic Activity of Labisia pumila (Kacip Fatimah) Stem and Root Extract on Human Blood 1 Student : Nur Batrisyia Binti Mat Ruzi Supervisor : Dr. Norhaida Che Azmi Introduction: Thrombolytic agents still have significant shortcomings, including the need for large doses to be maximally effective, limited fibrin specificity, and bleeding tendency. Objective: Hence, this research aims to investigate the anticoagulant and thrombolytic activity of Labisia pumila stem and root. Method: The methanolic extract of Labisia pumila stem and root was evaluated by measuring their effect on the prothrombin time (PT) and activated partial thromboplastin time (aPTT) tests. The thrombolytic activity was measured with clot lysis test on venous blood drawn from 10 healthy volunteers. Statistical analysis of the PT and aPTT results was performed using a Statistical Package for the Social Sciences (SPSS) version 27 by applying mean values using one way analysis of variance (ANOVA) and Dunnett’s post-hoc tests. Results: Based on the results obtained, Labisia pumila stem and root extract did not show significant prolonged anticoagulant activity in the PT test. The aPTT test showed increased anticoagulant activities through mean ± standard deviation (SD) as the extract concentrations were raised at 5 mg/mL (68.33 ± 4.06 sec), 10 mg/mL (82.33 ± 11.59 sec), and 20 mg/mL (89.67 ± 22.59 sec) respectively. The Thrombolytic Assay also depicted notable clot lysis activity with 20 mg/mL extract concentration through mean ± SD, 60.70% ± 20.25 and the p-value is significant compare to positive and negative controls (0.018, 0.005). Conclusion: This study suggests that L. pumila could be a potent source for developing anticoagulant and thrombolytic drugs to treat diseases related to blood clot disorder. Keywords: anticoagulant, thrombolytic, Labisia pumila, methanolic, aPTT, PT YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 66
In Vitro Study of Anticoagulant and Thrombolytic Activity of Murayya koenigii Extract on Human Blood 1 Student : Wan Arisha Dania binti Wan Mohd Adha Supervisor : Dr. Norhaida Binti Che Azmi Introduction: Abnormalities within coagulation cascade contributes to several vascular diseases: venous thromboembolism (VTE), pulmonary embolism (PE) and myocadiac ischemia. Anticoagulants and thrombolytics are common treatments, however due to the narrow therapeutic index, adverse effects are possible. Objective: Hence, this research focuses on herbal alternatives by investigating the anticoagulant and thrombolytic activity of Murraya koenigii methanolic extract using in vitro study. Method: Prothrombin time (PT) and activated partial thromboplastin time (aPTT) test were utilized to study the anticoagulant activity of M. koenigii methanolic extract at varied doses (5mg/mL, 10mg/mL, and 20mg/mL). Thrombolytic assay used fresh blood from 10 healthy volunteers and 20mg/mL M. koenigii methanolic extract. Clotting time and clot lysis (%) were analysed using ANOVA and T-Test. Results: The results yield significant results (p<0.05) for anticoagulant activity of M. koenigii methanolic extract utilising PT and aPTT test, with highest clotting time of 24.97s and 53.81s at 20mg/mL extract respectively. In contrast, thrombolytic activity of M. koenigii was insignificant as extract yield the lowest clot lysis (%) of 31.39% when compared with control group. Conclusion: M. koenigii do possess anticoagulant activity, however subtle thrombolytic effect was observed on human blood through in vitro study. Keywords: Murraya koenigii, anticoagulant, PT, aPTT, blood coagulation, thrombolytic YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 67
In Vitro Study of Anticoagulant and Thrombolytic Activity of Capsicum frutescens’ Extract on Human Blood 1 Student : Nur Jannah Binti Edy Murad Supervisor : Dr. Norhaida Binti Che Azmi Introduction: An unregulated blood coagulation caused excessive clotting, which then led to Thromboembolic diseases such as stroke, myocardial infarction, pulmonary embolism, and deep vein thrombosis (DVT). Numerous plants used in various systems of traditional medicine for the treatment of thromboembolic diseases demonstrated anticoagulant or antithrombotic activity. Capsicum frutescens, also known as bird’s eye chilli, chilli padi, Piri-Piri, and Hawaiian pepper, was used to treat arthritis, inflammation, cardiovascular diseases, and was also used as a pain reliever. However, its thrombolytic and/or anticoagulant properties were not sufficiently studied. Objective: This in vitro study aimed to explore the anticoagulant and thrombolytic activities of Capsicum frutescens’ methanolic extract. Method: Concentrations of 5mg/mL, 10mg/mL, and 20mg/mL were assessed for anticoagulation using prothrombin time (PT) and activated partial thromboplastin time (aPTT) tests, while thrombolytic activity was evaluated through clot lysis studies at 20mg/mL. Results: Significant anticoagulant activity was observed at 20mg/mL when compared to the negative control (P<0.05). At 5 and 10mg/mL, the plant extract exhibited some anticoagulant effect but there was no statistical significance (P>0.05). In the thrombolytic assay, substantial activity was noted at 20mg/mL compared to the negative control (P<0.05), although significance was not achieved against the positive control (P>0.05). Nonetheless, the observed significance in thrombolytic activity against the negative control underscores the extract's potential to lyse blood clots. Conclusion: In conclusion, our analysis reveals significant anticoagulant and thrombolytic activities in Capsicum frutescens’ , suggesting its potential therapeutic role in blood coagulation disorders. Keywords: Activated partial thromboplastin time, Anticoagulant activity, Capsicum frutescens, Methanolic extract, Prothrombin time, Thrombolytic activity, Thromboembolic diseases. YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 68
The Effects of Panax ginseng and Ophiocordyceps sinensis Supplementation on Spatial Learning and Memory Assessment in Animal Models Using Radial Arm Maze 1 Student : Nur Kamalia Ezza binti Bahrin Supervisor : Mdm. Nabilah Hanim Binti Mohd Sabri Introduction: The Radial Arm Maze is a well-known instrument for testing spatial cognitive ability in animal models. This study employed the Radial Arm Maze to thoroughly evaluate the effects of Panax ginseng and Ophiocordyceps sinensis supplementation on spatial learning and memory in animal models. Objective: The objective was to identify any differences in effects and determine whether one supplement produced higher cognitive benefits over the other. Method: In this study, 18 male Sprague-Dawley rats were categorized into three groups: control, ginseng, and cordyceps, with each consisting of six rats. Behavioral observations were performed using radial arms to evaluate changes in working and reference memory before supplementation with treatment. Over a 7-day period, all rats received a 0.5 mL solution via a daily force-feeding procedure. The ginseng group received a solution containing 18 mg of ginseng, while the cordyceps group was given 36 mg. Post supplementation assessments using the Radial Arm Maze (RAM) test were performed again. Results: This study's findings demonstrated a significant decline in working memory, as evidenced by reduced arm re-entries during the post-test across all groups. This observed reduction indicates a potential improvement in cognitive performance post supplementation. Furthermore, all groups demonstrated a decline in reference memory, as measured by the frequency of entry into baited arms, following supplementation. These findings suggest that the treatments have a beneficial effect on cognitive function, notably in the areas of working and reference memory, emphasizing the potential cognitive benefits of the experimental interventions. Conclusion: The findings suggest that there is a potential influence of Panax ginseng and Ophiocordyceps sinensis on spatial-learning and memory enhancement. However, it is noted that the length of supplementation may be a critical factor and further adjustments might yield more prominent results. Keywords: Radial Arm Maze, Panax ginseng, Ophiocordyceps sinensis, Sprague-Dawley rats YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 69
Synthesis of Sabinene Loaded Nanocellulosic Film as an Antimicrobial Wound Dressing Material 1 Student : Nur Liyana Afiera Binti Mohd Fuad Supervisor : Dr. Tong Woei Yenn Introduction: A large part of the world’s population has been impacted with chronic wounds. There has been up to 13 million people globally suffered from chronic wound annually. This issue has called for a thorough wound care where wound dressings have been the popularized in protecting chronic wounds from being infected. Objective: Despite the existence of various types of wound dressings, the limitation in every type of dressings is still greatly discussed. Sabinene, which is a type of monoterpene compound that can be extracted from various plants possess antimicrobial and antifungal properties have been used in developing sabinene loaded nanocellulosic film to be used as wound dressing material. Method: In this study, cellulose nanocrystal was successfully extracted from medical grade cotton fibres by acid hydrolysis. Then, the antimicrobial activity of the developed film was tested using agar diffusion assay, Hohenstein Challenge test and Wash Durability test. Results: The nanocellulosic film loaded with sabinene was white in colour, flexible and soft. The needle-like structures of nanocellulose fibres were observed by using field emission scanning electron microscope. Sabinene release test was performed and the results showed a slow and gradual release of sabinene from the nanocellulosic film in the span of 48 hours without any burst effect. The film also showed significant inhibitory activity on 2 Gram positive bacteria, 1 Gram negative bacteria and 2 yeasts. In Hohenstein challenge test, the film has shown remarkable bacteria reduction (99%) on all test microorganisms. In addition, even after 15 washes in Wash Durability Test, the film had high endurance. Conclusion: The results have stipulated the efficacy of sabinene loaded nanocellulosic film to be utilized as an antimicrobial wound dressing material. Further investigations should be done regarding the use of nanocellulose incorporated with other antimicrobial agent in the development of wound dressing. Keywords: Nanocellulose crystal, Sabinene, Cellulose, Wound dressing, Chronic wounds YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 70
The Comparison of Helminths DNA Detection between Conventional PCR and Real-Time PCR (RT-PCR) Using Modified DNA Extraction Method 1 Student : Nur Nabilah Binti Azman Supervisor : Dr. Mehru Nisha Co-Supervisor : Dr. Pang Jyh Chyang Introduction: Helminthic infections, including the nematode A. lumbricoides, are today known as a global threat, as 1.5 billion people worldwide are infected with higher vulnerabilities. Rapid and accurate detection of helminth infections is crucial for disease control, environmental monitoring, and surveillance. Due to insufficient gold standards, the detection has relied on insensitive microscopybased techniques. There is a low significance of specificity and sensitivity to detect intestinal helminths compared to molecular assays. PCR efforts can provide absolute quantification and simultaneously reduce helminth burdens. Since helminth eggs are hard to lyse, the physical egg lyses method could greatly help yield more DNA. Hence, this study aimed to compare conventional PCR and real-time PCR by a modified extraction method. Objective: To investigate DNA helminth detection between conventional PCR and real-time PCR via a modified DNA extraction method. Method: Random stool samples (n=20) with suspected helminths A. lumbricoides were investigated via microscopy and concentration techniques. The mechanical treatment of freeze-thaw and boiling-microwaving methods was used to disrupt helminth eggs. Next, DNA was extracted using a commercial kit. The extracted DNA was subjected to conventional PCR and realtime PCR screening. Results: Around 20 stool samples, which were positive in microscopy for Ascaris lumbricoides, were used for conventional PCR and realtime PCR. Meanwhile, 16 stool samples (80%) were evaluated as positive outcomes on DNA amplification by conventional PCR, and only five stool samples (25%) were successfully obtained by real-time PCR. Conclusion: Microscopy techniques were effective due to their simplicity and precision in helminth detection. Contrarily, molecular assays such as real-time PCR have explicitly high limitations in the quantification of cDNA due to the presence of possible contaminants in stool DNA. Keywords: conventional PCR, real-time PCR, soil-transmitted helminths, molecular, DNA amplification, DNA extraction YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 71
Effects Of Different Concentrations Of Bisphenol A (BPA) Exposure On Yeast Growth 1 Student : Nazmin Binti Mohamad Zaki Supervisor: Ts. Dr. Noor Arniwati Mat Daud Co-Supervisor : Mr. Ammarul Zaki Rahman Introduction: In recent years, concerns have risen about the potential environmental and health impacts of bisphenol A (BPA), a common chemical found in plastics and resins. While numerous studies have focused on its effects on animals and humans, there is a lack of comprehensive understanding regarding how BPA affects simpler biological organisms like yeast. Yeast, due to its cellular similarities with more complex organisms, can serve as a model for studying cellular responses to chemical exposures. Objective: This research objective is to investigate the effects of various concentrations of BPA on yeast growth, providing insights into the physiological impacts of BPA on yeast. Method: In this study, different concentrations of BPA (0.1 ppm and 0.3 ppm) were used to investigate the impact on the yeast growth by determining the optical densities of the yeast cells over a 4-hour period at 600 nm using a spectrophotometer. Results: The optical density of the control group increases in the first hour, indicating normal growth. This initial increase is due to the lag phase, where yeast cells adapt to their new environment and prepare for division. After the lag phase, they enter the exponential phase, where they multiply and reach their maximum growth rate. As they continue to grow and consume nutrients, they enter the stationary phase, where they maintain a constant optical density. In contrast, the BPA-exposed group experiences a temporary adaptive response, followed by BPA-induced toxicity, which negatively impacts yeast cell growth and proliferation over time. Conclusion: In summary, the use of optical density measurements via spectrophotometer to evaluate the toxicity of BPA on yeast growth appears to be a valid method as the changes in optical density over time provide insights into the growth dynamics of the yeast cells in response to BPA exposure. Keywords: Bisphenol A, yeast, viability, oxidative stress, control group, BPAexposed group YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 72
Protective Potential Effects of Curcumin on AspirinInduced Kidney Function in Rats 1 Student : Batrisyia Balqis Binti Mohd Asri Supervisor : Associate Professor Ts. Dr. Reezal Ishak Introduction: Aspirin is frequently used due to its well-established beneficial effects including treatment for prevention of cardiovascular disease (CVD), and protect against cancer when prescribed at preventative doses. However, excessive consumption of aspirin can cause hepatotoxicity, nephrotoxicity, and life threatening. Curcumin is a yellow-colored plant polyphenol has been shown to possess a wide spectrum of biological actions including its protection against drug-induced toxicity. Objective: Therefore, the current study aims to investigate the effect of curcumin extracted from turmeric (Curcuma longa) against aspirin nephrotoxicity in rats through determination of kidney function parameters. Method: To achieve this aim, twelve adult male Sprague Dawley rats were divided into equal three groups as following, Group I served as control, Group II served as positive control (Aspirin 300 mg/kg b.wt/orally), and group III (Aspirin 300 mg/kg b.wt/orally and curcumin 400 mg/kg b.wt/orally). After 14 days and at the end of experiment (after 28 days), blood were collected for biochemical, and histopathological analysis. Results: After a two weeks of observation in both untreated and treated aspirin groups, no significant changes were observed in the levels of serum creatinine and uric acid. However, after four weeks, a marked increase was noted in the untreated aspirin group, which was statistically significant. In addition, the histopathological examination also revealed a significant retraction of the glomerular tuft and loss of brush border. Interestingly, the administration of curcumin over a four weeks demonstrated a dose-dependent reduction in the elevated levels of kidney function biomarkers, thereby attenuating the harmful effects of aspirin. Conclusion: In conclusion, the obtained results indicated that curcumin exhibits protective effects against aspirin-induced renal toxicity through its antioxidant properties, particularly evident after four weeks of regular consumption. Additionally, this suggests that the observed changes in renal biomarkers take some time to manifest, becoming evident only after an extended duration of the study. Keywords: Aspirin, Curcumin, Nephrotoxicity, Creatinine, Uric acid. YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 73
The Effect of Soursop (Annona muricata) Leaves Extract on Blood Glucose and Pancreas Histology in Streptozotocin Induced Diabetic Rats 1 Student : Nur Uzma Damia Binti Mohd Nasir Supervisor : Mdm. Zainie Aboo Bakkar Introduction: Diabetes mellitus is a metabolic illness that involve chronic hyperglycemia triggered by either an insufficient production of insulin, resistance to insulin or both that may harm the central nervous system, resulting in neurodegeneration and many alterations in the brain’s structure and biological activities. Recent studies have shown that the leaves of Annona muricata can be used as an alternative traditional treatment in treating diabetes mellitus due to the presence of chemicals of acetogenins which provide various health benefits. Objective: The aim of this research is to study the effects of Annona muricata extract on blood glucose levels and brain histology in streptozotocin-induced diabetic rats. Method: A total of 16 male Sprague-Dawley rats used in this study were divided into 4 groups. Group 1 was the control, Group 2 Annona muricata treated group, Group 3 was streptozotocin with treated Annona muricata group and Group 4 was untreated Streptozotocin group. All of the animals were monitored for two weeks. Daily force feeding of 200 mg/kg Annona muricata was administered to group 3 and group 4 for two weeks. On day 15, the brain was harvested to observe any pathology or anomalies that affected the brain. Results: The data obtained were analysed with descriptive analysis using one-way annova. The result of the body weight that affects Annona muricata indicates that, followed the administration of STZ, there was no statistically significant (p<0.05) difference observed amongst the four animal groups. In the meantime, following the administration of STZ, the blood glucose result was considerably (p<0.05) different. During the brief 14-day course of treatment, Annona muricata's brain histology shows some encouraging results. Conclusion: In conclusion, the study demonstrated that Annona muricata possesses anti-hyperglycemic activities. Keywords: Annona muricata, Diabetes Mellitus, Streptozotocin, Blood Glucose Level, Brain YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 74
The Effect of Annona muricata (Soursop) Leaves Extract on Wound Healing and Skin Histology in StreptozotocinInduced Diabetic Rats 1 Student : Nur Wafa Sorfina Binti Ahmad Shukri Supervisor : Mdm. Zainie Aboo Bakkar Co-Supervisor : Dr. Ahmad Najib Hasan Introduction: Diabetes mellitus (DM) is a chronic metabolic disorder characterized by elevated blood glucose levels, known as hyperglycemia. It occurs when the body cannot produce enough insulin or effectively use the insulin significantly impacting the normal process of wound healing. Objective: Recognizing the challenges posed by impaired wound healing in diabetes, this study aims to evaluate the effectiveness of Annona muricata leaves extract on wound healing and skin histology in streptozotocin-induced diabetic rats. Method: The Annona muricata leaves were soaked in 70% ethanol solvents and were extracted using a rotary evaporator at 45 ℃ with 150rpm. A total of 12 male Sprague Dawley rats were used in this study by randomly divided into four types of groups which are (A) diabetic control, (B) vehicle control, (C) oral treated with Annona muricata (200mg/kg once a day) and (D) topical treated with 10% w/w of Annona muricata ointment. Diabetics were induced in all groups by a single intraperitoneal injection of 65mg/kg streptozotocin dissolved in sterile water injection. Every rat was assessed for blood glucose level, body weight, wound closure percentage, and skin histological changes for 14 days. Results: The results in this study indicate that there were no significant differences (p>0.05) recorded in fasting blood glucose levels and body weight measurements of treated diabetic rats and non-treated diabetic rats. Meanwhile, the macroscopic and microscopic analysis of the diabetic wound has demonstrated a complete epidermal regeneration with the presence of matured granulation tissue and less inflammatory cells. Thus, revealed that both oral and topical treatments with Annona muricata have led to a significant improvement in wound closure percentages compared to the control diabetic rats’ group (p<0.001). Conclusion: These findings have shown that there are therapeutic benefits of Annona Muricata of both topical and oral treatment intervention in enhancing the healing process. Keywords: Diabetes Mellitus, Annona muricata, streptozotocin, wound healing, wound closure percentage YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 75
The Effect of the Soursop Leaves (Annona muricata) on Blood Glucose Level and Liver Histology in Streptozotocin-Induced Diabetic Rats. 1 Student : Wan Athirah Binti Wan Shukri Supervisor : Mdm. Zainie Aboo Bakkar Introduction: Diabetes mellitus is one of the major chronic metabolic diseases that can give more effect on our body. Previous study stated that Annona muricata can inhibit α-glucosidase and α-amylase activity which prevent an increase blood glucose level and commonly used to treat diabetes traditionally. Objective: Thus, this study aims to investigate the effect of the Annona muricata leaves extract on liver histology in streptozotocin induced diabetic rats. Method: 16 male Spargue-Dawley rats were randomly divided into four groups (n=4 per group): Control group received no intervention (A); Normal group treated with Annona muricata 50mg/kg for 14 days (B); Diabetic group treated with Annona muricata using the same dosage through the oral force feeding for 14 days; Untreated diabetic group (D).The blood glucose, body weight and liver histology were observed for each rat of the groups after 14 days. Data were presented as mean ± SD and statistically analysed by ANOVA. Results: The results showed a significant increase (p<0.05) in blood glucose level after the streptozotocin (STZ) administration at days 6 and days 15. There was not statistically significant (p<0.05) different in the body weight after STZ administration. The liver histology shows the involvement of the hepatocytes with features of inflammation in diabetic group. Conclusion: These findings suggested that Annona muricata treatment can be taken for a prolonged period for more effective results. Keywords: Diabetes, Streptozotocin, Annona muricata, Blood Glucose Level, Liver YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 76
The Effect of Annona muricata Extract (Soursop Leaves) on Blood Glucose Levels and Kidney Histology on Streptozotocin (Stz) – Induced Diabetic Rats 1 Student : Nurin Zulaikha Binti Zain Supervisor : Mdm. Zainie Aboo Bakkar Introduction: Diabetes mellitus (DM) is a chronic metabolic condition characterized by hyperglycemia caused by deficiencies in insulin production, action, or both. Several studies have looked at the possible benefits of natural products to treat diabetic symptoms and complications, one of which being Annona muricata, often known as soursop. Objective: The purpose of this study is to examine whether Annona muricata extract (AME) will have an impact on blood glucose levels and kidney histology in streptozotocin-induced diabetic rats. Method: 16 Sprague-Dawley rats were divided into four groups of rats (n= 4 per group); Group A served as “Normal control” received no intervention; Group B served as “Normal rats with AME treatment”; Group C served as “AME treated diabetic rats”; Group D served as “Untreated diabetic rats”. Diabetes was induced in Groups C and Group D by a single intraperitoneal injection of STZ (65 mg/kg) that was dissolved in distilled water. Group B and C were treated with AME 200 mg/kg daily for 14 days via oral route. After 14 days, all the rats were sacrificed, and organs were harvested. The data obtained were analysed using SPSS Statistic version 27. Results: The result showed a mean body weight of 331.75±23.91g, 313.25±38.98g, 249.00±21.21g, and 238.00±108.89g respectively for the control group, normal rat with AME treatment group, treated diabetic group and untreated diabetic group. The mean blood glucose level of 5.78±0.37 mmol/L, 6.35±0.58 mmol/L, 19.95±0.21 mmol/L, and 13.10±11.17 mmol/L for the control group, normal rats with AME treatment group, treated diabetic groups and untreated diabetic group respectively. ANOVA analysis showed a significant difference (p<0.05) between the blood glucose concentrations of untreated diabetic groups and AME treated diabetic groups. Conclusion: The findings from this study suggest that ethanol leaf extract of Annona muricata demonstrated anti-diabetic effect in rats and may be beneficial in the treatment of diabetes. Keywords: Keywords: Diabetes mellitus, Annona muricata extract, Blood glucose level, Kidney histology, Body weight, Rats YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 77
The Association between Sweet Dessert Intake and Fasting Blood Glucose Levels Among Health Science and Engineering Science Student 1 Student : Nurul Hidayah Binti Norul Azman Supervisor : Mdm. Nur Shukriyah Mohammad Hazir Co-Supervisor : Dr. Azizah Mat Hussin Introduction: Food and emotions are strongly related they can cause our bodies to experience a wide range of feelings. It has been discovered that while some foods, such as those high in fat, carbohydrates, and sugar, tend to make individuals feel bad, most people associate sweet desserts and meals high in sugar with making them feel good and content. Objective: Blood glucose levels are monitored, and the amount of sugar that the blood is carrying at any given time is calculated. This sugar can be consumed by people through their eating styles. Method: A cross-sectional study design will be conducted at UniKL Mestech Kajang and UniKL MFI Bangi in Hulu Langat, Selangor, Malaysia. The questionnaire was used to collect data on the reading blood glucose level demographic data, Frequency of Food, Physical activity, General Health, and Lifestyle. Data will be analyzed by using Statistical Package for the Social Sciences (SPSS). Results: The response rate for the questionnaire was 96% (N=100). From the target high glucose readings are higher among engineering students at 20.8% compared to health science students at 14.6% In contrast, health science students show a relatively lower percentage of high glucose readings at 14.6%. Conclusion: The study found the pattern of sweet desert food and fasting blood sugar profile of study participants health science and engineering sciences student Keywords: Sweet dessert, glucose, Health Science (MESTECH), Engineering Science (MFI) YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 78
Antibacterial Effects of Cymbopogon citratus & Mentha piperita Ethanolic Extracts Mixture on Streptoccocus pyogenes and Staphylococcus aureus 1 Student : Nurul Huda Izzah Binti Ahmad Khalil Supervisor : Mrs. Nur Shukriyah Mohamad Hazir Introduction: Pharyngitis (strep throat) is an inflammation of pharynx that is caused by Group A Streptococcal (GAS). However, as we move forward to 2023, the rise of multidrug resistance microbes threatens the therapeutic efficacy of commonly used antibiotics. It is draw to the inference that Cymbopogon citratus (lemongrass) & Mentha piperita (peppermint) have a strong antibacterial activity, however the state of antibacterial activity of the plant mixtures is still unexplored. Objective: Thus, the significance of this study is to investigate the antibacterial effects of lemongrass and peppermint mixture on Streptococcus pyogenes and S. aureus through in vitro study. Method: The ethanol extract of lemongrass and peppermint were prepared with the concentrations of 200mg/ml respectively for the antibacterial testing. Agar well diffusion method was done and the zone inhibition for each of the bacteria upon experiment was recorded. A serial dilution method for Minimum Inhibitory Concentration and Minimum Bactericidal Concentration were also done towards ethanolic extracts of lemongrass, peppermint, and the mixtures. Results: Results showed lemongrass ethanol extracts was more effective against Streptococcus pyogenes than S. aureus where the inhibition zone for both microbes is ranging from 15-20mm and 0-9mm respectively where this might be due to variances in the morphological structures of these organisms. The inhibition zone of peppermint extracts for S. aureus and Streptococcus pyogenes is ranging from 12-18mm and 14-20m respectively. The inhibition zone of the mixtures towards S. aureus and Streptococcus pyogenes is ranging from 7-11mm and 14-19mm. According to the results obtained, S. aureus and Streptococcus pyogenes were found to be more resistance against the plant mixtures compared to the peppermint ethanolic extracts alone. The statistical analysis showed that there is statistically significant between positive control and the plant mixtures which p-value was <0.05. Conclusion: Therapeutic potency of lemongrass and peppermint ethanolic extracts was proved as they do exhibit antimicrobial effects on S. aureus and Streptococcus pyogenes. However, the mixtures of both extracts do not produce a better antimicrobial effect. Keywords: Cymbopogon citratus, Mentha piperita, Staphylococcus aureus, Streptococcus pyogenes, Mean Inhibition Zone, Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 79
Effects of Carrot (Daucus carota subsp. Sativus) Extract on Candida albicans and Streptococcus pyogenes 1 Student : Nurul Iman Jamiah Binti Mohamed Ishak Supervisor : Mdm. Nur Shukriyah Mohamad Hazir Introduction: Carrot (Daucus carota subsp. Sativus) exhibit antimicrobial and antifungicidal properties against various pathogens. The exploration of natural and sustainable alternatives creates a possible therapeutic agent for Streptococcus pyogenes and Candida albicans to address infections and human diseases towards people who suffered unwanted side effects and allergic reaction of drugs. Objective: Hence, the aim of our study is to explore the potential of carrot extract against the growth of Streptococcus pyogenes and Candida albicans. Method: Carrot constituents were extracted using 70% ethanol and tested for anti-susceptibility activity against Streptococcus pyogenes and Candida albicans at varying concentrations (31.25-500 mg/mL). Agar well diffusion method and broth microdilution for detecting zone of inhibition and minimum inhibitory concentration (MIC) were used. Results: The carrot extract against the two organisms were observed with no zone of inhibition for anti-susceptibility test. Optical Density (OD) reading in MIC for Candida albicans and Streptococcus pyogenes also showed negative result. Conclusion: Carrot extract does not show inhibitory effect as compared to control (p<0.05). Both organisms do not display inhibition towards carrot extract. Keywords: carrot, Streptococcus pyogenes, Candida albicans, zone of inhibition, minimum inhibitory concentration. YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 80
The Effect of Lawsonia inermis and Calendula officinalis Combined Formulation on Laceration Wound Healing 1 Student : Nurul Najwa Binti Nasri Supervisor : Mdm. Nur Shukriyah Mohamad Hazir Introduction: Wound healing is a complex physiological process that involves various cellular and molecular events. The exploration of natural remedies for wound care has garnered considerable interest due to their potential therapeutic benefits. Among these remedies, henna (Lawsonia inermis) and calendula (Calendula officinalis) have been individually recognized for their traditional uses in promoting skin health and wound healing. Objective: This study aimed to investigate the effect of Lawsonia inermis and Calendula officinalis combined formulation on laceration wound healing. Method: Rats with standardized laceration wounds were divided into three experimental, positive, and negative control groups. The experimental groups were received 5% ointment topical application of the henna, calendula, and combined formulation. The positive control group were treated with a commercial wound care product while the negative control group were treated using petroleum jelly. This study was span for 14 days and monitored the visual examination, wound closure rate, and histological analysis. Results: Results reveal that the combined formulation group demonstrates a wound-healing effect on laceration wounds in rats. By day 14 post-treatment, it was observed increased wound contraction and reduced wound length through macroscopic examination and histological analyses showed inflammatory cells, hair follicle formations, and collagen depositions. Statistical results showed that there was no significant difference between combined formulation group with a positive group. Conclusion: Combined formulation of henna and calendula give effect on laceration wound healing. Thus, this finding may serve as cost-effective alternative for wound care. Keywords: wound healing, henna, calendula, rat, combined formulation, laceration wound YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 81
Comparison Between Garlic (Allium sativum) And Ginger (Zingiber officinale) Aqueous Extract on The Growth of Candida spp. 1 Student : Nurulhuda Binti Mohd Suhairi Supervisor : Mdm. Nur Shukriyah Mohamad Hazir Introduction: Fungal infections, Candida albicans, are a serious health issue, especially in immunocompromised patients. Known to be present in 30% to 60% of healthy individuals, these species are the most frequent hospitalizations in the general American population. Non-albicans Candida (NAC) species become more pressing due to resistance to antifungal medications. Medicinal plants like garlic and ginger have long been utilized for medicinal and antimicrobial properties, but research on their use against pathogens is limited. Objective: This study aims to compare the effectiveness of garlic and ginger extracts on the growth of Candida spp. Method: The methods used were the agar well diffusion method, minimum inhibitory concentration, and minimum fungicidal concentration. Results: The results showed the mean zone of inhibition of Candida albicans for 100%, 50% and 25% of aqueous garlic extract concentration is 62.67±2.517 mm, 54.67±5.033 mm and 45.67±3.215 mm while for Candida tropicalis is 63.33±2.517 mm, 55.33±2.517 mm and 47.67±4.041 mm. No inhibition zone observed for aqueous ginger concentration. MIC and MFC of aqueous garlic extract for both Candida spp were found to be effective at 12.5%. The statistical analysis showed that when compared with positive control and aqueous garlic extract was statistically significant which p-value was <0.05. This study found that aqueous garlic extracts showed higher inhibition zones than clotrimazole. Conclusion: Allicin, a bioactive compound in garlic, inhibits Candida albicans growth by damaging fungi’s cell wall. However, aqueous ginger extracts showed no effect against Candida albicans and Candida tropicalis. From this research, the results obtained confirm that garlic shows potential antifungal against Candida spp compared to ginger while ginger shows no potential antifungal. Keywords: garlic, ginger, Candida spp, Candida albicans, Candida tropicalis, antifungal, plants, clotrimazole YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 82
The Synergistic Effect of Camellia sinensis with Gentamicin Against Methicillin-Resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa 1 Student : Nur Nisa Amalin binti Mohd Nazri Supervisor : Mdm. Noor Eszrezad Juferi Introduction: The skin plays an essential function in preserving general health by acting as a barrier against microbial attackers. However, certain pathogens pose a formidable challenge to this protective shield, leading to skin infections of significant concern. Methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa are particularly notable offenders, exhibiting a concerning capacity to avoid conventional therapeutic approaches and cultivate resistant infections. Camellia sinensis (green tea) plant leaves have a lot of benefits, such as having a wide spectrum of antioxidant, anti-inflammatory, anticarcinogenic, and antibacterial properties against numerous pathogens. Objective: The aim of this study is to investigate the synergistic effect of Camellia sinensis with Gentamicin against Methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. Method: The antimicrobial sensitivity test of Camellia sinensis extract at 100 mg/mL was determined on a disc diffusion assay. Then, the Minimum Inhibitory Concentration (MIC) of Camellia sinensis extract was determined by a broth microdilution assay, which then proceeded with the Minimum Bactericidal Concentration (MBC) test. The synergistic antimicrobial activity of Camellia sinensis extract with Gentamicin was assessed using disc diffusion by comparing ZOI alone. Results: The MICs of Camellia sinensis extract alone for MRSA and Pseudomonas aeruginosa were 6.25 mg/mL and 25.00 mg/mL, respectively. Next, the Minimum Bactericidal Concentration (MBC) of Camellia sinensis extract was 50.00 mg/ml, where the MBC result was higher than the MIC. However, the results for the MIC and MBC of Gentamicin were 10 mcg. Conclusion: The findings conclude that the combination of Camellia sinensis with Gentamicin does exhibit synergistic interaction against MRSA and Pseudomonas aeruginosa. Keywords: Synergistic effect, Synergism, Camellia sinensis, Gentamicin, MRSA, Pseudomonas aeruginosa YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 83
The Synergistic Effect of Camellia sinensis with Ciprofloxacin Against Methicillin-Resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa 1 Student : Nur Syahirah Binti Khairul Nizam Supervisor : Mdm. Noor Eszrezad Juferi Introduction: Food is a basic need for human survival. Consumption of contaminated food by microbes like Methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa leads to increased cases of Foodborne diseases (FBD) resulting in high morbidity and mortality rates across the world. Emergence of antimicrobial resistance (AMR) also causes difficulty in treating diseases as the microbes develop resistance over the past year causing alternative treatments using plants either alone or in combination with common antibiotics to curb the issue. Camellia sinensis or green tea (GT) was selected for this study due to its high bioactive compound contents like catechin responsible for exhibiting antimicrobial properties. Objective: This study aims to investigate the synergistic effect of Camellia sinensis with Ciprofloxacin against Methicillinresistant Staphylococcus aureus and Pseudomonas aeruginosa. Method: By using disc diffusion assay, the antimicrobial activity of methanolic GT extract at 100 mg/ml and Ciprofloxacin on MRSA and Pseudomonas aeruginosa was assessed by measuring the zone of inhibition (ZOI). Then, the Minimum Inhibitory Concentration (MIC) of GT at concentration from 50 to 0.78 mg/ml was determined by broth microdilution method which then proceeded with Minimum Bactericidal Concentration (MBC) test. The synergistic antimicrobial activity of GT with Ciprofloxacin was then assessed using disc diffusion by comparing ZOI alone. Results: The MIC of GT alone on MRSA was 6.25 mg/ml was lower compared to MIC of GT alone on Pseudomonas aeruginosa were 25.00 mg/ml. Both bacteria recorded a similar value of MBC at 50.00 mg/ml. The MIC and MBC result of Ciprofloxacin recorded were 5 µg. The combination of GT extract with Ciprofloxacin displayed a significant synergism effect by increased ZOI against MRSA and Pseudomonas aeruginosa. Conclusion: The findings conclude that the combination of Camellia sinensis with Ciprofloxacin does exhibits synergism interaction against MRSA and Pseudomonas aeruginosa. Keywords: Camellia sinensis, green tea, ciprofloxacin, MRSA, Pseudomonas aeruginosa, synergism, synergistic effect, YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 84
1 Bachelor of Environmental Health (Honours) ABSTRACTS YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 85
Awareness Levels in Fresh Produce Preferences Between Organic and Commercial Sources Among Public in Semenyih, Selangor 1 Student : Aqina Syafira Binti Azman Supervisor : Dr. Nurul Shyha Md Yatim Co-Supervisor : Ts. Dr. Noor Arniwati Mat Daud Introduction: Pesticides, chemical substances employed to combat organisms that harm crops, including insects and rodents, are commonly used in farming. Most fresh produce originates from farms where pesticides, especially chemical ones, are applied. The use of pesticides can impact human health, leading to acute effects like dizziness and chronic effects such as asthma. Objective: This cross-sectional study aimed to evaluate the awareness levels regarding the effects of pesticide application on fresh produce on human health. Additionally, it sought to explore public preferences for either commercial or organic fresh produce. Method: The study, conducted in Semenyih, Selangor, involved 336 participants (90.81% of the public population). A Google form was utilised as a tool, and the questionnaire comprised three sections. The survey was distributed online and in grocery markets. Data were analysed using descriptive and Spearman correlation tests in SPSS version 27. Results: The study encompassed 336 participants in Semenyih, Selangor. Results revealed that 72.9% of respondents exhibited a high level of awareness regarding pesticides and their health impacts. Additionally, 64.3% preferred commercial fresh produce, with freshness (84.8%) being the primary criterion for purchase. Among those selecting commercial produce, 28.3% cited ease of availability as the main reason, while 35.7% of organic produce advocates emphasized the lower health risk (26.2%). Significantly, a difference was observed between public preferences and awareness levels (p-value=0.005). Conclusion: Notably, a significant difference exists between public preferences and awareness levels. Nevertheless, future research could benefit from employing experimental methods to determine pesticide concentrations in fresh produce. Keywords: Pesticides, Fresh produces, Awareness, Public preference YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 86
Pesticide Knowledge and Safety Practices Among Smallholder Farmers in Muar, Johor 1 Student : Arisya Hanim binti Sharol Hisam Supervisor : Dr. Nurul Shyha Md Yatim Introduction: Globally, there are 759 chemical and organic pesticides utilized in agriculture and the medical sector. Pesticides in agriculture confer numerous benefits, including the control of agricultural pests, thereby ensuring food safety. The absence of pesticides could result in an annual loss of up to 30% in agricultural production. In response to the limitations of traditional plantation systems in curbing pest infections, Malaysia's agriculture heavily relies on the common practice of pesticide use. Despite the undeniable advantages of pesticides, production workers and farm labourers, face a heightened risk of pesticide poisoning, with 5 million cases reported annually. Objective: This crosssectional study aims to assess the knowledge, attitude, and practices (KAP) regarding pesticide handling among 234 smallholder farmers in Muar, Johor. Method: The structured self-administered questionnaire comprises four sections: Section A (sociodemographic), Section B (knowledge of pesticide handling), Section C (attitude toward pesticide handling), and Section D (safety practices in pesticide handling). Results: The findings reveal that smallholder farmers in Muar, Johor exhibit a high level of knowledge (50%) and a positive attitude (50.5%) towards pesticide handling. However, the practical implementation of safety measures remains at a lower level (45.3%). Significantly, there is an association between knowledge levels and the number of years spent in the agriculture sector (x2=13.39; p-value=0.001). Additionally, this study investigates the connection between attitude and practice levels concerning sociodemographic factors among smallholder farmers. The results indicate significant associations between both attitude and practice levels with sociodemographic factors, except for the type of plantation, which lacks a significant association with practice level (x2=5.56; p-value=0.135). Conclusion: In conclusion, this study emphasizes the imperative for smallholder farmers in Muar, Johor, to possess not only knowledge and a positive attitude but also to implement sound work practices to mitigate the risk of pesticide poisoning and serious health effects. Keywords: Pesticide handling, Smallholder farmers, Knowledge, Attitude, Practices, Muar, Johor, Cross-sectional study. YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 87
Identification of Risk Factors that Contribute to Food Premise Rating at Parliament Segambut, Kuala Lumpur 1 Student : Muhammad Akmal Shah Bin Saim Supervisor : Mr Zakaria Hamid Co-Supervisor : Ts. Arvind Bala Krishnan Introduction: Foodborne diseases, such as food poisoning, typhoid, and cholera, are a major concern in Malaysia, with an estimated 30% of individuals becoming ill each year due to water and food diseases. High rates are attributed to poor hand hygiene practices and lack of public knowledge. The increasing demand for food services and the rise in food festival events also contribute to foodborne illnesses. The government is implementing medication and awareness programs to address these issues, and the Ministry of Health is appointing enforcement authorities to monitor and improve food safety. Objective: The purpose of this study was to identify the risk factors that contribute to food premise rating at Parliament Segambut. Method: The food premises grading ‘A’ and ‘B’ are included in the study, the food premises grading ‘C’ are excluded in the study. In compliance with the Food Hygiene Regulations of 2009, a cross-sectional descriptive study employing a quantitative approach was conducted on one hundred (N=100) food establishments, and the results were analysed. Results: Overall, the risk factor contribute to non-A grading compliant are use of aprons and head covering, cleanliness of food container, displaying food ingredients in a perfect way and protected from contamination, Using a grease trap (grease trap) and it well function and maintenance of grease trap. In conclusion, the p-value was less than the standard (p<0.05). Thus, there is a significant difference the component of risk factor and food premises grading. Conclusion: Therefore, the food industry should be concerned about this issue to ensure that their employees follow the law and to prevent future outbreaks of food poisoning linked to poor hygiene and food safety. Keywords: Risk factor, Food premises grading YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 88
Association Between Level of Knowledge towards Food Safety and Personal Hygiene Practices Among Food Handlers in Segambut 1 Student : Nor Asiah Binti Mohd Zain Supervisor : Mr. Zakaria Hamid Introduction: This research aims to investigate the knowledge towards food safety and personal hygiene practices among food handlers in Segambut. The research will assess the food handlers' understanding of food safety regulations, their attitudes towards implementing proper food safety measures, and their dayto-day practices in maintaining food hygiene and safety standards. Food can become contaminated by shippers, processors, distributors, and other parties throughout the process. Incorrect food handling procedures are the primary cause of food contamination, which can lead to various kinds of food-borne illnesses. Objective: The general objective of this research is to determine the association between level of knowledge towards food safety and hygiene and personal hygiene practices among food handlers in Segambut, Kuala Lumpur. Method: A quantitative research had been done to 80 food handlers across various food establishments. All the data then run for statistical analysis to identify their association by conducting Chi Square Test and descriptive using SPSS. Results: The respondent’s knowledge level was majority in a moderate level (68%) while the level of personal hygiene practices is not much different which also in moderate level (76%) respectively. There is no significant association between level of knowledge and personal hygiene practices since the p value (0.085) is more than α. Conclusion: Language barrier is the most common issue faced by food handlers in Segambut. Authorities could develop efficient strategies to improve communication together with their knowledge and personal hygiene practices. Keywords: Food safety, Food hygiene, Food handler, Language barrier YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 89
The Association Between the Level of Knowledge of Food Handlers and Level of Hygiene of Food Premises in Segambut 1 Student : Nur Fatihah Binti Abdul Razak Supervisor : Mr Zakaria Hamid Introduction: Food poisoning and many other foodborne illness is a serious issue due that may cost lives. Unhygienic preparation of food and unclean condition of food premises may be one of the factor for a foodborne illness to be easily transmitted. Food handlers need to have good knowledge in food safety as they hold accountability to maintain good hygiene practice and cleanliness of food premises. Objective: The objectives of this study is to identify the association between level of knowledge of food handlers and the level of hygiene of food premises. Method: 80 food premises from two grades, A and B are identified in Segambut, Kuala Lumpur. The data on socio-demographic and knowledge of food handlers of those food premises in Segambut are collected through questionnaires. Results: The result indicated that 57.5% of the respondents has high knowledge of food handling and safety. However, there is no significant association between the education background and involved years in food industry and level of hygiene of food premises (p=0.427, p=0.291). Meanwhile, there is significant association between the presence of food handling training of food handlers and the level of hygiene of food premises with p-value of <0.001. There is significant association between the level of knowledge of food handlers and level of hygiene of food premises in Segambut, with p-value of 0.007. Conclusion: From this study, it can be concluded that food handlers need to have good knowledge in food handling and safety in order to sustain a hygienic condition of food premises. Keywords: Food handlers, Food premises, Food hygiene, Knowledge YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 90
Evaluation of The Relationship Between Fractional Exhaled Nitric Oxide (FeNO) with House Environment in Urban and Suburban Areas 1 Student : Nurin Nadhirah Binti Abdullah Supervisor : Dr. Khairul Nizam Mohd Isa Introduction: Fractional exhaled nitric oxide (FeNO) levels were reliable indicators of airway inflammation, and the home environment is a crucial field of research since it contributes to the rising prevalence of respiratory illnesses. Objective: This research aimed to measure the FeNO levels in exhaled breath samples and evaluate the conditions or environment of the house in urban and suburban areas. Method: This comparative cross-sectional study design involved 470 secondary students (270 urban, 200 suburban) aged 14 from 8 schools located in urban and suburban areas in Hulu Langat district, Selangor. The respiratory symptoms related to the house characteristics were assessed using a standardized questionnaire from the International Study of Asthma and Allergies in Childhood (ISAAC). Then, FeNO levels were measured using the NIOXVERO® 12-1200 airway inflammation monitoring device, and atopy classification was determined by allergy skin prick tests. Multiple logistic regression analyses were used to determine the association. Results: Overall, atopy was found in 59.4% and 40.6% of students in urban and suburban, respectively. About 90.9% of suburban students reside in wooden houses and 58.2% of urban students had floor deterioration in the past 12 months. The nasal infection showed a significant association with the occurrence of water problems (OR=2.14, 95% CI=1.24-3.71) and mouldy floors, walls, or ceilings (OR=3.03, 95% CI=1.22-7.56) in urban and correlation with floor renovation (OR=5.15, 95% CI=1.65-16.10) in suburban. Water-leaking mould shows strong associations with weekly rhinitis (OR=8.60, 95% CI=1.52-48.81) in suburban and wheezing (OR=8.29, 95% CI=3.65-18.86) in urban. A significant association between elevated FeNO more than 35 ppb with weekly ocular symptoms in urban (OR=4.30, 95% CI=1.32-14.03) and with wheezing in suburban (OR=5.99, 95% CI=2.24-15.99). Conclusion: There was a significant relationship between Fractional Exhaled Nitric Oxide (FeNO) with house environment in urban and suburban areas. Keywords: FeNO, House environments, Urban, Suburban YOUNGSCIENTISTSYMPOSIUMUniKLMESTECH2023 91