Institut Perubatan & Pergigian Termaju - Tesis

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Anion Exchanger 1 Gene (AE1) Single Nucleotide Polymorphism (SNP) Analysis Among Malays With Distal Renal Tubular Acidosis (DRTA) And Southeast Asian Ovalocytosis (SAO)
(2018-08)
Anion Exchanger 1 (AE1) encoding the erythroid and kidney isoforms of AE1 protein is located on chromosome 17. Distal renal tubular acidosis (dRTA) is characterized as its inability to acidify urine pH caused by AE1 polymorphisms. Southeast Asian Ovalocytosis (SAO) is a hereditary red blood cell abnormality caused by 27bp deletions in AE1. dRTA and SAO is highly prevalent in Southeast Asian countries. Polymorphism in AE1 is known to cause both SAO and dRTA associated with 27bp deletions which is still unclear.
Anti-Inflammatory Effects Of Mesenchymal Stem Cells Derived Extracellular Vesicle In Rat Model Of Chronic Obstructive Pulmonary Disease
(2022-01)
Noridzzaida Binti Ridzuan
This study aimed to assess the anti-inflammatory effects of human umbilical cord mesenchymal stem cell (hUC-MSC) derived EV in a rat model of COPD. Human UC-MSC-EV were isolated and characterized by using transmission electron microscope, western blot, and nanoparticle tracking analysis
Antiproliferative And Apoptosis Effects Of Clinacanthus Nutans Leaf Extract On Breast Cancer Cell (Mcf7)
(2023-02)
Noor Zafirah Binti Ismail
The objective of this study was to determine the antiproliferative and apoptotic effects of C. nutans leaf extract on the breast cancer cell line, Michigan Cancer Foundation-7 (MCF7).
Association Analysis Of Single Nucleotide Polymorphism Of Opioid Dependence Genes Among Malay Males In Malaysia
(2018-09)
Nagaya, Devaki
Drug addiction is a chronic and relapsing disorder is associated with genetics. There are many studies have been reported the roles of single nucleotide polymorphisms (SNPs) with drug dependence and among those, twelve (12) candidate genes that were associated with drug dependence will be. investigated in this study. The candidate genes are OPRD1 (Delta Opioid Receptor), OPR.K1 (Kappa Opioid Receptor), COMT (Catechol — O-Methyltransferase gene), PDYN (Prodynorphin), DRD4 (Dopamine receptor D4), ABCB1(P glycoprotein), DUSP (Dual Specificity Phosphatase 27) and rs 10494334. The goal of this study was to determine the frequencies of these SNPs rsl042114, rs702764, rs 199774, rs 1022563, rs910080, rs737866, rs 10494334, rsl800955, rsl 128503, rsl045642, and rs2032582 and to study their association with opioid dependence in Malay males in Malaysian population.
Biocompatibility And Toxicity Studies On Heterogeneous Tio2-Zno Polymer Nanocomposite With Enhanced Bacterial Activity
(2022-07)
Nor Hazliana Harun
Healthcare-associated infections (HAIs) are a major safety concern globally as they contribute to mortality rates amongst patients due to pathogens from direct contact with a contaminated biomedical polymer surface from the indwelling or implanted medical devices. Polymer nanocomposites have become a promising solution for HAIs owing to reactive oxygen species (ROS) and free radicals’ mode of action. In the first section, this work revealed the bacteriostatic and bactericidal potentials of TiO2–ZnO polymer nanocomposite films against HAI pathogens, including multidrug-resistant (MDR) and non-MDR strains. In the second section, the initial analysis of the in vitro bio-interaction responses on fibroblast and blood cell line models showed signs of cell membrane integrity disturbance, which might be due to free radicals’ activities, such as the release of intracellular ROS and Zn ions (Zn2+) during the initial cellular adaptation process on the TiO2–ZnO polymer nanocomposite film. Molecular studies revealed that the cell–polymer nanocomposite film interaction possibly triggered the oxidative stress and pro-inflammatory mechanisms through the principal cascades of Nuclear Factor-κB. Further analysis found that cells could maintain the viability and clonogenic potential and were involved in the anti-apoptosis pathway. Findings suggested the transitory oxidative stress responses of polymer nanocomposite films towards treated cells and not harmful to the cells.
Characterisation Of Biogenic Silver Nanoparticles-garcinia Atroviridis Toxicological Effects In Normal And Cancer Human Lung Cells.
(2024-09)
Muhamad, Musthahimah
Silver nanoparticles (AgNPs) synthesized from plants are increasingly used across various industries, including the pharmaceutical and food sectors. This widespread use has raised concerns about their safety profiles. This study aimed to examine the toxic effects and mechanisms of biogenic silver nanoparticles (AgNPs-GA) synthesized using Garcinia atroviridis leaf extract on human lung cancer cells (A549) and normal lung cells (BEAS-2B) in vitro. Cytotoxicity was evaluated using the MTT assay to determine the IC50 values of AgNPs-GA in both cell types. The IC50 values were applied to treat the respective cells for evaluating reactive oxygen species (ROS) production, apoptosis, DNA damage, silver ion levels, and cellular uptake using CM-H2DCFDA assay, DAPI and AOPI double staining, comet assay, ICP-OES, and TEM, respectively. The mRNA expression and metabolic profiles were further assessed using quantitative real-time PCR and untargeted metabolomics based on ultra-high-performance liquid chromatography-Orbitrap mass spectrometry (UHPLC-Orbitrap-MS), respectively. The results showed that AgNPs-GA inhibited 50% of the growth in A549 and BEAS-2B cells, with IC50 values ranging from 20–28 μg/ml and 12–35 μg/ml, respectively. The selective index (SI) values of AgNPs-GA were below 3, classifying it as a non-selective cytotoxic agent, as it induced general toxicity in both cancerous and non-cancerous cell types. AgNPs-GA were primarily internalized and deposited within lysosomes, nucleus, and mitochondria, leading to an increased release of ionic silver (Ag+) inside both cells.
Development And Validation Of Online Preconcentration Capillary Electrophoresis Methods For The Determination Of Selected Pharmaceutical Compounds In Aqueous Matrices
(2022-01)
Semail, Nadhiratul-Farihin
In this study, several preconcentration strategies in capillary electrophoresis (CE) were developed to improve the sensitivity of detection of pharmaceutical compound in real aqueous matrices. This is due to the core challenge to develop simple and sensitive analytical method for trace analysis study of selected analytes (anticancer, antibiotic and non-steroidal anti-inflammatory drugs) are very limited. Higher interference level from complex matrix (biological, food and water) with analytical instrument poor detection capability directed to development of online preconcentration technique. The first technique termed sequential injection mode stacking was developed for the separation and determination of anticancer agent 5-fluorouracil (5-FU) and its metabolite namely, 5-fluoro-2-deoxyuridine (5-FdUrd) in human plasma obtained from cancer patients. In this technique, large sample volume was introduced through electrokinetic injection followed by hydrodynamic injection. Under the optimized experimental conditions, 614- and 643- fold and 782- and 803- fold sensitivity improvement was obtained for both 5-FU and its metabolite when compared with normal hydrodynamic and electrokinetic injection alone, respectively.
Development Of Antibacterial Chitosan-Based Wound Dressing Incorporated With Zno/Cuo Nanoparticles Using Calotropis Gigantea
(2023-06)
G Ambarasan A/L Govindasamy
In this work, ZnO/CuO nanocomposites incorporated in chitosan (Cs) based biopolymer were studied for physicochemical, antibacterial, cytocompatibility and wound healing profiles. This ZnO/CuO-Cs nanocomposites were obtained by freeze-thawing process and characterized by XRD, SEM, TEM, EDAX, FTIR, AAS, ICP and UV-vis spectroscopy.
Development Of Lead-free Tungsten Carbide-based Polymeric Bricks As Gamma Radiation Shielding
(2022-06)
Abualroos, Nadin J D
Lead bricks are the most widely used form of gamma ray shields in diagnostic radiology, radiotherapy, and nuclear medicine departments. However, lead is heavy, toxic, and dangerous to human health. Polymer-based radiation shields can be utilised in radiation shielding field due to their excellent physical and chemical properties, ease of fabrication, low manufacturing costs, and toughness. Also, their radiation shielding properties can be improved by using high atomic number fillers. The purpose of this research was to develop a lead-free tungsten carbide-based polymeric bricks as gamma radiation shielding. The fabrication of tungsten carbide-based polymeric composites involved mixing tungsten carbide powder and tungsten carbide cobalt powder with epoxy resin at different weight percentages (60, 65, 70, 75, 80, 85, 90) of fillers. Also, addition of bismuth oxide and barium sulphate powder to tungsten carbide-based epoxy matrix were studied. The physical, morphological, chemical, mechanical, and photon shielding properties were investigated for all the fabricated composites. The most optimum powder epoxy combination among all samples were 85 and 90 weight percentage of fillers. The microstructural analysis indicated that the fillers distributed uniformly within the epoxy matrix. Some agglomerations were observed for composites having more than 75 wt% of filler. Fourier transform infrared spectroscopy was used to study the chemical properties of the fabricated composites. FTIR absorption spectrum of each sample analysed in the range of 600-4000 cm-1. The results obtained did not show any spectral change for all samples.
Elucidation Of The Role Of Long Noncoding Rna Kcnma1-As2 In Colorectal Cancer
(2024-07)
Miao, Xinzhi
The aberrant expression of long noncoding RNAs (lncRNAs) is frequently documented in colorectal cancer (CRC). This anomalous expression is associated with tumourigenesis by promoting detrimental biological behaviours in tumour cells, including proliferation, apoptosis, cell cycle progression, and migration. A growing body of evidence suggests that lncRNAs play pivotal roles in tumour regulation at both molecular and cellular dimensions. Some lncRNAs function as oncogenes and exhibit increased expression, while others act as tumour suppressors and show reduced levels in cancerous tissues. Furthermore, lncRNAs can regulate processes by targeting miRNAs, serving as competing endogenous RNAs. This study seeks to discover novel lncRNA entities and comprehensively examine and validate their roles and underlying molecular mechanisms in CRC. Tissue samples, both tumourous and non-tumourous, from three CRC patients were submitted for sequencing. The sequencing results indicated 148 up-regulated and 146 down-regulated lncRNAs. Following expression validation in samples from ten patients and four CRC cell lines, LINC00668 was identified as an up-regulated lncRNA. An siRNA knockdown was performed on LINC00668 to assess if altering its impact on CRC cell functions. The MTT assay was used to measure cell proliferation. The Annexin-V/PI apoptosis assay was performed to determine cell apoptosis. To analyse the cell cycle, a cell cycle assay was conducted, and a wound healing assay was utilised to evaluate cell migration. However, no significant effects on cell proliferation or apoptosis were observed post-knockdown of LINC00668, leading to its exclusion from subsequent research.
Feasibility Of Complementary Metal Oxide Semiconductor (Cmos)-Based Time-Of-Flight (Tof) Image Sensors In Conjunction With Kilovoltage (Kv) Cone Beam Computed Tomography (Cbct) For Real-Time Tumour Tracking In Radiotherapy
(2022-03)
Lim, Siew Yong @ Siew Eng
Respiratory induced tumour movement is one of the main causes of error in thoracic radiotherapy. The tumour movement may results in sub-optimal delivery of radiation to the target during treatment. The objective of this research is to investigate the feasibility of two Complementary Metal Oxide Semiconductor (CMOS)-based Time-of-Flight (ToF) image sensors in conjunction with kilovoltage (kV) cone-beam computed tomography (CBCT) imaging system for real-time tumour tracking in radiotherapy. A dynamic phantom was designed to simulate the periodic breathing motion of patient. Two ToF image sensors (Argos3D P320 & P330, BECOM Electronics, Austria) of different pixel array size and type of illumination system were employed.
Green Synthesis Of Silver Nanoparticles From Water-Soluble Polysaccharides Of Clinacanthus Nutans Leaf: Extraction, Characterisation, Optimisation And Antibacterial Studies
(2023-07)
Devasvaran, Kogilavanee
C. nutans has gained much attention by researchers due to its pharmacotherapeutic properties. However, the extraction method of C. nutans leaf crude polysaccharide (CNP), CNP synthesising silver nanoparticles (AgNP) and the subsequent antibacterial properties as a DNA synthesis inhibitor, has not been studied. Hence, this research aims to employ a green synthetic method to produce AgNP and assess its potential application as an antibacterial agent. Five extraction methods of CNP were employed, namely hot water (HW_CNP), ultrasound-assisted (UL_CNP), freeze-thaw (FT_CNP), alkaline (ALK_CNP) and acid (ACD_CNP). All five CNP were optimised and characterised, and their compositions were analysed. Three CNP were then selected to synthesise ACD_AgNP, ALK_AgNP, and UL_AgNP, respectively, where ALK_AgNP and UL_AgNP were optimally selected based on their synthesis conditions and their toxicity in brine shrimp was quantified. The antibacterial and potential DNA synthesis inhibition mechanism of action (MOA) of ALK_AgNP was further quantified. The results of CNP showed that ACD_CNP and FT_CNP obtained the highest and lowest extraction yield at 14.85 ± 0.25% and 4.16 ± 0.13%, respectively.
Immunoproteomic Analysis And Serodiagnosis Of Salmonella Enteretica Typhi And Invasive Non-Typhoidal Salmonella Infections Using Differentially Extracted Whole Cell Bacterial Protein
(2023-04)
Abdul Lateef Khan, Khairunnisa
Typhoid fever is a potentially fatal infectious disease caused by Salmonella Typhi. The disease is endemic in developing countries, but alarming number of cases linked to domestic sources have been reported in industrialized countries. The clinical presentation of typhoid fever is highly variable and often overlap with other acute undifferentiated febrile illnesses (AUFI). Hence, accurate identification of isolates is important for effective epidemiological intervention. The development of rapid, costeffective diagnostic test is also crucial for early life saving treatment. Therefore, comparative analysis was performed to characterize protein profiles of two differentially extracted Salmonella serovars by sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE). Separation of protein into three different extractions revealed discriminative protein profiles between the two strains which provided an effective adjunct that can be used as a tool for protein serotyping. A standardized protocol of the dot Enzyme Immunoassay (dot EIA) with differentially extracted antigens derived from S.Typhi and S.spp was established for definitive and differential diagnosis of typhoid fever. The performance of the test was encouraging with high sensitivity and specificity. Following the successful development of 'TYPHOIDYNE μSpot M-Ag Arrays, the antigenic components of the differentially extracted proteins were further characterized by western blot (WB) assay.
In Silico And In Vivo Analysis Of Otu Deubiquitinases Otub1, Otub2 And Otulin Protein-protein Interactions
(2023-09)
Zulkifli, Nur Wahida
In this post-genomic era, proteomic and interactomic data are important sources for understanding the molecular basis of cell functional diversity. Research in these fields is still progressing, with at least 90% of proteins annotated so far and many more interactions to be uncovered to complete the proteome and interactome maps. The purpose of this study is to contribute to the expansion of human interactome data by focusing on the deubiquitinating enzymes (DUBs) protein-protein interaction (PPI) using several in silico and in vivo approaches. First, a DUBs interactome was built on Cytoscape ver 3.9.1 using data from IMEx database and analysed using various graph theory algorithms. The DUBs interactome consisted of 3,406 nodes and 4,982 edges, whereas a cancer protein subnetwork extracted from the interactome revealed that DUBs are significant in cancer biology. Concluding the interactome study, OTUB1, one of the proteins with strong network characteristics, along with its closest homologue OTUB2, and the newest family member OTULIN, were selected for yeast two-hybrid (Y2H) screening against human cDNA library derived from HEK293. Y2H bait and cDNA library prey vectors were generated using Gateway technology, first in donor vector TOPO/pDONR222 before being shuffled into destination vector pDEST32/22. The bait vector is sequence-verified, and the prey vector is of sufficient quality and quantity to represent human cDNA with total clones of 6.8×106 cfu. Both vectors were transformed into MaV203 S. cerevisiae strain for Y2H screening to identify putative interacting proteins
Investigation Of Time-Of-Flight (TOF) Imaging System During Deep Inspiration Breath-Hold Radiotherapy (DIBH-RT) For Left Breast Cancer Patients
(2022-04)
Auwal, Abu Bakar
Deep inspiration breath-hold radiotherapy (DIBH-RT) pushes the heart away from treatment field thereby reducing the cardiac dose by over 50%. However, poor reproducibility and stability of the breath-hold during DIBH-RT treatment delivery could jeopardise the treatment success. The purpose of this work was to study the application of a Time-of-Flight (ToF) imaging system for monitoring breath-hold during DIBH-RT. Argos P330 3D ToF Camera (Bluetechnix Austria) was characterised by imaging an ISOCube phantom to assess the ToF measurement stability in conjunction with CBCT. Subsequently, the ToF imaging was performed on a total of 13 DIBH-RT patients during setup verification with CBCT imaging. Chest surface displacements were estimated from each modality and compared to validate the ToF measurement.
Investigation On Antiproliferative Mechanisms Of Alstonia Angustilobasilver Nanoparticles In Skin Squamous Cell Carcinoma
(2023-08)
Ab Rahim, Nurhidayah
Traditional methods of nanoparticles synthesis frequently use toxics chemicals and substantial energy, which may have detrimental effects on the environment and human health. Recently, plant-based synthesis of nanoparticles (AgNPs) has emerged as a promising green synthesis method. Herein, we reported the green synthesis of AgNPs using the aqueous extract of A. angustiloba leaves and assessed its antiproliferative mechanisms. Initially, the physiochemical properties of A. angustiloba-AgNPs were characterised using UV–Vis spectrophotometry, FT-IR, FESEM, EDX, TEM, XRD and zeta sizer analyses. The cytotoxicity of A. angustiloba-AgNPs was examined by MTT assays against the A431 cancer cell line. The morphology of the treated cells was examined using fluorescence microscopy and the antiproliferative mechanisms of the nanoparticles in A431 cells were investigated by annexin-FITC/propidium iodide (PI) staining and DNA cell cycle analysis using flow cytometry. The intracellular ROS levels were measured using a commercially available kit. The expression of apoptosis and cell cycle-related proteins was determined by Western blotting.
Investigation On Cancer Chemopreventive Activity Of Fabricated Tq Encapsulated Plga-Pf68 Nanoparticles On Selected Breast Cancer Cell Lines
(2022-02)
Nurul Shahfiza Binti Noor
Drug resistance is remains as a major obstacle in breast cancer therapy. This has attracted considerable interest to overcome drug resistance with bioactive compounds. Free thymoquinone (TQ) has been extensively studied for its anticancer properties. In the clinical setting, its therapeutic potential has not been explored due to its volatile nature. Therefore, the study aimed to develop TQ polymeric nanoparticles and evaluate its effects on drug resistant breast cancer cells.
Isolation And Characterization Of Aptamers Against Leptospira Lipl32
(2023-06)
Yeoh Tzi Shien
In this study, both DNA and RNA aptamers were developed against LipL32. LepRapt-11 was shown to be the most potent RNA aptamer, with dissociation constant and limit of detection of 350 ± 47.45 and 100 nM as determined by direct Enzyme-linked Aptasorbent Assay (ELASA), respectively.
Molecular Toxico-Pathology Responses Of EUDRILUS EUGENIAE TO 2- AMINO-l-METHYL-6-PHENYLIMIDAZO[4,5- bJPYRIDINE (PhIP) AND 2-AMINO-3,8- DIMETHYLIMIDAZO [4,5-f]QUINOXALINE (MeIQx)
(2019-01)
Balogun, Wasiu Gbolahan
The carcinogenic risks of highly heated proteinaceous foods are at large associated with the formation of heterocyclic amines (HCAs), evidence in their substantial presence in the biota. The earthworm Eiidrilus eugeniae (E. eugeniae) is a terrestrial ubiquitous animal, susceptible to toxicity from environmental contamination, and a suitable model for understanding the mechanism of toxic agents such as HCAs. Thus, this study elucidated the vulnerability of E. eugeniae to HCAs induced toxicity using a modified OECD TG207 approach.
Optimisation Of Induced Pluripotent Stem Cells Derivation From Human Cord Blood And Its Application For Cardiac Regeneration In Cryoinjured Rat Model
(2024-08)
Universiti Sains Malaysia
Cardiomyocyte therapy was considered the only option to repopulate myocardial loss, but challenges existed in achieving effective replenishment of functional cardiomyocytes that could translate into clinical benefits. This study aimed to examine the use and function of pre-epicardial cells in cardiomyocyte therapy in vivo. Cryopreserved cord blood was used to generate human-induced pluripotent stem cells (hiPSCs) to harness the ‘youthfulness’ of foetal cells. The generated hiPSCs were also used to produce cardiomyocytes (CMs) and pre-epicardial cells (PECs) with high differentiation efficiency using established protocols. To confirm the effects of PECs on cardiomyocytes, both differentiated CMs and PECs were co-cultured for 6 days, and the proliferation and role of Hippo signaling were assessed. PECs (1x10⁶ cells/100 μl) were also combined with CMs (1x10⁶ cells/100 μl) to treat cryoinjured rat hearts after 1 month. Cardiac functions were assessed using Millar Pressure Volume Loop cardiac hemodynamics analysis. Differences between groups were considered significant when p < 0.05 using ANOVA. Frozen CD34 cells were successfully reprogrammed into hiPSCs with consistent expression of pluripotent markers. The generated hiPSCs demonstrated multilineage differentiation ability, forming cardiomyocytes, dopamine neurons, and early endodermal cells.

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