Institut Perubatan & Pergigian Termaju - Tesis

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Optimization Of Immunoillstochemistry: Testing For Mlhl And Msh2 Proteins In Colon Tissue
(2005-05)
Muhsain, Siti Nur Fadzilah
Microsatellite instability (MSI) accounts for the development of about 15% of all colorectal cancers. It is characterized by mutations in mismatch repair (MMR) genes that render cells unable to detect errors during DNA replication. Mutations in the MMR genes, MLHl and MSH2 are responsible for the majority of CRC with MSI. These MMR proteins can be detected by immunohistochemistry (lllC). IHC has proved sensitive and specific when compared to molecular analysis of MSI. Nevertheless, generally, there is no published literature as yet on studies of MSI in Malaysia, and specifically using lliC. Before any lliC staining procedure can be done, it has to be optimized. In this study, attempts were made to optimize rnc staining of MLHl and MSH2 proteins using polyclonal rabbit antibody by Biovision on formalin-fixed paraffm-embedded sections of normal colon, taken from resected margins of colectomy specimens. Either Vectastain Elite ABC Kit or DAKO En Vision Kit was used as secondary detection system. The staining procedure was done using different dilutions of the primary antibodies and incubated for different durations at different temperatures. Antigen retrieval procedures were introduced using citrate buffer pH 6 or Tris-EDTA buffer pH 9. The results showed that incubation of sections at room temperature using different dilutions of antibody failed to produce any nuclear staining. Staining with DAKO En Vision was still unsatisfactory, although background staining was reduced. Introduction of antigen retrieval procedure using microwave did not improve results. When incubated at 4°C, non-specific background staining was produced.
Determination Of Epstein-barr Virus In Gastrointestinal Lymphomas (With Special Reference To Non-hodgkin's Lymphoma Of Small Intestine): Study On Malaysian Patients.
(2005-05)
Yunos, Asmah Md
Non-Hodgkin's lymphoma (NHL) is ranked twelfth among all cancers world-wide, in which it is more prevalent in males compared to females. The aim of the present study was to determine the presence and distribution of Epstein-Barr virus (EBV) in non-Hodgkin's lymphomas (NHL) of the gastrointestinal tract (GIT) tissue samples obtained from 25 Malaysian patients diagnosed clinically and histopathologically. The gastrointestinal lymphomas analysed in the present study was divided into NHL of the ileocecal (9 cases), NHL of the small intestine (7 cases), NHL of the stomach (5 cases), NHL of the abdomen (1 case), NHL of the wedge ulcer (1 case), NHL of rectum fotic (1 case) and finally NHL of the hemicolon (1 case). The presence of EBV in all of the above mentioned tissue samples, after formalin fixation and paraffin embedding was carried out by using conventional in situ hybridization technology. Two out of 25 cases of NHL of the GIT analysed in the present study, demonstrated positive signals for EBV/EBER. In the first positive case, EBV/EBER signals were located in transformed lymphocytes in the serosa layer of the small intestine (14.3%). In the second EBV positive case, EBV/EBER signals were located in diffuse B-cell lymphomas of the ileocecal (11.1 %). In conclusion, these finding demonstrate a rare association between EBV and lower gastrointestinal lymphomas (excluding stomach).
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.
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.
Nanoencapsulation Of Thiolated Sodium Alginate: Synthesis, Drug Release And Cellular Uptake Studies
(2018-11)
Oral drug delivery has becoming the most preferred route of administration for chemotherapeutic drugs, however its application is limited due to lack of drug targeting specificity to the localised cancerous cells but not to the normal cells. Polymeric nanoparticulate drug delivery is a potential solution to deliver hydrophobic drugs specifically to colon, thus causing less side effects.
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.
Effects Of Human Umbilical Cord-Derived Mesenchymal Stem Cells On Human Corneal Epithelial Cells’ Growth
(2019-12)
Ex vivo cultivated corneal limbal stem cell (LSC) transplantation is the contemporary therapy for comeal epithelial regeneration. The success rate of this therapy is dependent on the cell culture conditions and the carrier system. Human umbilical cord mesenchymal stem cells (hUC-MSCs) has regenerative ability and are rich with paracrine factors. Cells cultivated in spherical scaffolds could provide a similar in vivo environment to the cell niche and may positively alter the stem cells properties. This study aims to improve the cellular quality of the comeal epithelial transplantation graft using a robust cell population method. Human telomere modified comeal epithelial cells (hTCECs) were co-cultured in direct and indirect cell culture with hUC-MSCs. Characterisation of corneal markers p63, ABCG2, ABCB5, and differentiation marker K3 were performed.
Generation Of RNA Aptamer Against Progesterone Receptor DNA Binding Domain And Its Potential In Diagnostic And Therapeutic Applications In Breast Cancer
(2021)
Ravinderan, Presela
Aptamers are a new class of molecular recognition element that exhibit high binding affinity and specificity against the target. In this study, an RNA aptamer was generated against Progesterone Receptor DNA binding domain (PR DBD) that acts as a diagnostic biomarker and potential therapeutic target for breast cancer. Firstly, PR DBD gene was amplified and isolated from the total RNA of MCF-7 cells and cloned into pET15b plasmid. Protein expression and native purification was performed using E. coli Rosetta 2(DE3)pLysS bacterial strain. The identity of the protein was confirmed by western blot using polyclonal anti-PR antibody and by protein sequencing using MALDI-TOF/TOF mass spectrometry. The purified protein was subjected to SELEX in order to isolate RNA aptamer. A total of eight SELEX cycles were executed and the resulting nucleic acid pool from cycle 8 showed enrichment against PR DBD. The cycle 8 nucleic acid pool was sequenced using direct sequencing and crush and soak elution-based sequencing methods. Both sequencing methods revealed the presence of three different classes of sequences, with one class termed, PRapt-3 showed the strongest binding against PR DBD. The dissociation constant of PRapt-3 RNA aptamer was estimated at 380 nM ± 35 nM. PRapt-3 was successfully used to develop aptamer-based diagnostic assays such as ALISA, aptamer-based dot blot, aptamer-based western blot, aptacytostaining and aptahistostaining. PRapt-3 detected PR DBD in direct ALISA with a LOD of 69.44 nM. In the aptamer-based dot blot assay, PRapt-3 detected up to 6.25 pmol of PR DBD. PRapt-3 also successfully detected recombinant PR DBD and endogenous PR DBD in MCF-7 and HeLa cells on aptamer-based western blot assay. PRapt-3 demonstrated nuclear staining in aptacytostaining and with better penetration in aptahistostaining using the formalin fixed paraffin embedded breast cancer cell and tissue blocks. Apart from diagnostic application, the functionality of the PRapt-3 RNA aptamer was also investigated in the therapeutic applications. First, the antagonistic property of the PRapt-3 was investigated on the cell cycle analysis. Flow cytometry analysis showed that PRapt-3 RNA reduced the number of cells in S phase of MCF-7 cell cycle while reciprocally increased the percentage of cells in G0/G1. PRapt-3 can also act as a promising apoptotic inducing agent. In this study, it was found that PRapt-3 downregulated the anti-apoptotic gene, BCL-2 and upregulated the pro-apoptotic gene, BAX. PRapt-3 mitigated expression of the proliferationrelated genes. The expression level of proliferation-related genes, PI3K and AKT were reduced by PRapt-3 RNA. The downregulation of these genes showed that PRapt-3 was successfully employed to antagonize the interaction between PR and progesterone. In this entirety, PRapt-3 is a promising diagnostic and therapeutic agent.
Accuracy Of 131i Activity Quantification And Dosimetry In Thyroid Phantom Using 3d-image Based Spect/CT
(2021)
Mohd Yahya, Mohd Hizwan
Quantitative imaging gives a better understanding of the distribution of radioactivity that can be used in the estimation of absorbed dose in the specific target organ. The goal of this study was to obtain the accuracy of 131I activity quantification and dosimetry in thyroid uptake phantom using 3-dimensional (3D) SPECT/CT imaging. In this study, a series of SPECT/CT acquisitions of the thyroid uptake was performed. Images of different voxel sizes 64, 128, and 256 with several iterations numbers were reconstructed using ordered-subsets expectation maximization (OSEM). The quantification of the activity in the phantom was determined using a Medical Image Data Examiner (AMIDE) software based on the mean, maximum, 0.7 maximum and 0.6 maximum value. The cumulated activity in the phantom was determined by the calculating area under the curve (AUC) in the time-activity curve. The results showed that the mean value of 128 and 256 voxel show a better result with the error obtained between 4.1% and 18.2% whereas for the maximum value, it gives an overestimate count with the error up to 78%. The cumulated activity derived from SPECT/CT showed an error margin of 15%. The dosimetry result of organ absorbed dose using 3D imaged based dosimetry was approximately 16.1% lower than the absorbed dose measure using Optical Stimulated Luminescence (OSL) dosimeter. This research shows that the 3D image based SPECT/CT with good quantification accuracy capable of estimating organ absorbed dose in131I therapy.
The Effectiveness Of Applied Muscle Tension In Reducing Vasovagal Reaction Among Young Blood Donors In Kelantan
(2021-05)
Alam Faizli, Afif
Background: The vasovagal reaction (VVR) incidence is the most common adverse donor reaction poses a major risk to the donor's safety and a disincentive for repeat donation. This study's main objective is to determine the efficacy of applied muscle tension (AMT) in reducing the VVR incidence among young blood donors in Kelantan. Methods: A prospective comparative interventional study was conducted among 306 young blood donors that were equally assigned to AMT and non-AMT groups. Donors were assessed with Blood Donation Reactions Inventory at 30 minutes and 48 hours post blood donation. The subjects will be followed-up on whether they make at least one donation attempt in the next six months. Results: Out of 306 young blood donors, the incidence of VVR was 24.8%. The AMT group showed a significant reduction in the VVR incidence rate compared to the non-AMT group (p<0.001). There was no significant increment of donor return rate among the AMT group than nonAMT group upon follow-up in six months. In addition, female (p=0.002), low body weight (p=0.002), low estimated blood volume (p=0.033) and low systolic blood pressure (p=0.005), are important predictors of VVR. Conclusions: This study showed that AMT was an effective strategy to reduce the VVR incidence among young blood donors in Kelantan. Implementation of AMT is recommended to donors with high risk of developing VVR
A Five Year (2013-2017) Retrospective Study On The Data Of Laboratory Testing For Biocompatibility Of Medical Device Implant Registered With Malaysia Medical Device Authority
(2021-11)
Sunthara Murthi A/L Anamalai
The study focused on the implantable medical devices which require most number of biocompatibility data. Data collection form reflecting the MeDC@St data entry pages is prepared to manually retrieve the deposited information for registration between July 2013 to December 2017, amounting to 1925 registration with some exceeds 100 pages content per registration.
Dual-thresholding based sinogram interpolation To reduce metal artefact In Computed tomography imaging
(2021-11)
Mohamad sobri, Nurul Fathin
Metal artefacts can degrade the image quality of computed tomography(CT)images which lead to difficulties and errors in diagnostic analysis. Metal artefactsappear as bright and dark streaks around the metal object. It is caused bybeamhardening effects, photons starvation, non-linear partial volume, photonscatter, undersampling and inappropriate correction algorithm during reconstructionofimage. The aim of this study is to improve the sinogram interpolation techniqueformetal artefacts reduction (MAR) in CT images. The MAR algorithmof this studywas developed using MATLAB platform. The first step of the algorithmwas theacquisition of virtual sinogram from the CT image using Radon transformfunction. Then, the thresholding technique is used to detect and isolate the metal part withinthe sinogram of the CT image. Two interpolation methods, cubic spline (CS) andLaplace interpolation (LI) were applied to replace the missing sinogramdata. Theinterpolated sinogram was then reconstructed using backprojection (byinverseRadon function) and finally the thresholded and interpolated images were fusedtoproduce corrected image. A total of seven phantom and six clinical CTimages withdifferent artefacts severity were selected for MAR correction. Qualitativeandquantitative evaluation of the corrected CT images was performed to evaluatetheeffectiveness of the proposed MAR technique. The findings showed metal artefact was significantly reduced by LI method (p-value = 0.02) as compared to CSmethod(p-value = 0.17). From quantitative analysis, the artefact index (AI) for LI-correctedimages is lower (mean AI = 66.07), but CS-corrected images yielded higher AI values (mean AI = 141.35) than the original images (mean AI = 99.31). The resultsfrom qualitative analysis showed LI-corrected images received higher scores (meanscore ± SD of 1.75 ± 1.22) from radiologist compared to original image (1.08±1.08)and CS-corrected images (1.33 ± 1.23). In conclusion, LI method workmoreeffectively in artefact reduction on both phantom and clinical images of different degrees of artefact severity.
Effect Of Nipa (Nypa Fruticans Wurmb.) Vinegar On The Inhibition Of Intestinal Glucose Absorption
(2021-12)
Farjana Yasmin
Nipa palm vinegar is a traditional vinegar widely used in Southeast Asia as a condiment and a home remedy for several ailments. Previous studies have reported the antihyperglycemic activity of aqueous extract of nipa palm vinegar. In this study, the effects of consuming nipa palm vinegar on postprandial glucose levels were scrutinized.
Dual-Thresholding Based Sinogram Interpolation To Reduce Metal Artefact In Computed Tomography Imaging
(2021-12)
Nurul Fathin Binti Mohamad Sobri
Metal artefacts can degrade the image quality of computed tomography (CT) images which lead to difficulties and errors in diagnostic analysis. Metal artefacts appear as bright and dark streaks around the metal object. It is caused by beam hardening effects, photons starvation, non-linear partial volume, photon scatter, undersampling and inappropriate correction algorithm during reconstruction of image. The aim of this study is to improve the sinogram interpolation technique for metal artefacts reduction (MAR) in CT images.
In Silico Selection And Validation Of Dna Aptamer Against Progesterone Receptor Dna Binding Domain
(2022)
Tholasi Nadhan, Navien
Progesterone receptor plays an important role in the progression of breast cancer. Currently, antibody-based Immunohistochemistry is used in pathological assessment of PR levels for the detection of breast cancer. The shortcomings associated with antibodies pave the path to use aptamers as the alternatives. Aptamers are single-stranded DNA or RNA oligonucleotides generated by SELEX that are capable of binding to their cognate target molecules with high affinity and specificity based on their unique structural folding capacity.The tediousness and rigor associated with certain steps of the conventional SELEX intensify the efforts to select DNA aptamers using in silico-docking approach. That said, we report an in silico selection and validation of DNA aptamer to the progesterone receptor DNA binding domain (PR DBD) using ssDNA sequences derived from human progesterone response elements (PREs).Firstly, a library of sixty-four different nearnative ssDNA analogs of the corresponding PRE sequences was designed and subjected to secondary and tertiary structural determination. After that, docking between the ssDNA tertiary structures with the PR DBD was carried out using PatchDock. The sequence with the highest docking score was chosen as the aptamer candidate and further validated by in vitro direct ELASA.Among the candidates, we selected the ssDNA sequence (PRDBDapt17; 5′- AGAACAGCGTGTTCT -3′), which showed the highest docking scores of 11334 as a promising PR DBD binding aptamer. In addition, the PRDBDapt17 detected recombinant PR DBD in direct ELASA with a limit of detection of 3.91 nM. The dissociation constant was estimated at 366.6 nM. Therefore, PRDBDapt17 is a potential aptamer that can be used in the diagnosis of breast cancer.
Anti-inflammatory Effects Of Mesenchymal Stem Cells Derived Extracellular Vesicle In Rat Model Of Chronic Obstructive Pulmonary Disease
(2022-01)
Ridzuan Noridzzaida
Chronic obstructive pulmonary disease (COPD) is a chronic lung disease characterized by progressive airflow obstruction associated with chronic inflammation. The current treatment strategies are focusing on improving the symptoms and quality of life but do not provide cure for the underlying caused. Recently mesenchymal stem cells (MSC)-derived extracellular vesicles (EV) is actively being investigated as a potential source of new cell-free based therapy for COPD due to it’s ability to ameliorate inflammation, however no research has been conducted to study the anti-inflammatory effects of MSC-EV in COPD model.Thus, 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. Male sprague dawley rats (n=66) age 8-9 weeks were divided into 11 groups; Naïve, Cigarette Smoke (CS), Self-healing (SH), treatment groups (CS-hUC-MSC-EV, CS-hUC-MSC, CS-hUC-MSC-CM), vehicle groups (culture media alone (MD), and phosphate buffered saline (PBS)), and control (C) group (C-hUC-MSC-EV, C-hUC-MSC, and C-hUC-MSC-conditioned media (CM)). Five groups (CS, SH, CS-hUC-MSC, CS-hUC-MSC-EV, and CS-hUC-MSC-CM) were exposed to CS from 3 cigarettes for approximately 15 minutes per session, 2 times a day at 2 hours interval, 7 days a week, for 12 weeks. Meanwhile, Naïve, and control groups were left to breathe normal air. The treatments (hUC-MSC, hUC-MSC-EV, and hUC-MSC-CM) and PBS and MD were administered at week 13. Naïve and injury group were euthanized at week 13, while treatment groups, vehicle groups, and self-healing group were euthanized at week 15. Lungs from all groups were then subjected to histological analysis by using hematoxylin and eosin (H&E) staining, alcian blue-periodic acid Schiff (AB-PAS) staining, immunofluorescence staining, and microarray analysis. Increased lymphocytes count, inflammation in peribronchial and perivascular area, as well as parenchyma area, increased goblet cells count, increased emphysema, and increased p65 expression were observed in CS group as compared to Naïve group.Self-healing for two weeks did not reduce the inflammation in peribronchial and perivascular area, as well as parenchyma area. Self-healing for two weeks also did not reduce goblet cells count, emphysema, and p65 expression. In treatment groups, reduction of inflammation in peribronchial and perivascular area, as well as parenchyma area, reduced goblet cells count, and emphysema, reduced p65 expression were observed as compared to CS and SH groups. Meanwhile, the treatments did not induce inflammation or increased goblet cells count, and did not induced emphysema in rat exposed to normal air. Microarray analysis showed regulation of COPD related pathways and genes in CS, hUC-MSC-EV, hUC-MSC groups. hUC-MSC-EV, and hUC-MSC significantly regulating many genes expression including NFKB1, MAPK1, MAP2K1, JUN, PRKCZ, and P65. In conclusion, hUC-MSC-EV effectively ameliorating the CS induced inflammation and could potentially serve as a new cell-free based therapy for the treatment of COPD.
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.
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
Proteomic Analyses Of Streblus Asper Root Extract On Cervical Cancer
(2022-02)
Roslan, Muhammad Nabil Fikri
Cervical cancer is the third most prevalent cancer in women. Despite recent advances in cancer treatment, there were 569,847 incidents and 311,365 deaths worldwide in 2018. The current treatment options such as radiotherapy and chemotherapy have been typically found to cause adverse side effects that hinder the treatment efficacy. In contrast, natural products have been found to provide better treatment efficacy. Therefore, this study was conducted to explore the potential of one natural product known as Streblus asper root extract to be an alternative prospect to the current treatment practise.
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.

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