Institut Penyelidikan Perubatan Molekul - Tesis
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Browsing Institut Penyelidikan Perubatan Molekul - Tesis by Type "master thesis"
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- PublicationAntiaging Activity Of Polyphenol Rich Calophyllum Inophyllum Fruit Extract In Aging Mutants Of S. Cerevisiae(2022-12)S. Venugopal, KavilashaCalophyllum inophyllum L. is an important medicinal plant with many ethnomedicinal values and exhibited significant anti-oxidant activity. Hence, this study was conducted to determine the in vitro anti-aging property of C. inophyllum fruit extract (CIFE) against Saccharomyces cerevisiae BY611 Strain with the molecular mechanism of the anti-aging process. The anti-aging effect of CIFE against the yeast was evaluated through chronological lifespan assay, anti-oxidative stress assays, real-time quantitative polymerase chain reaction (RT-PCR) to study the regulation of superoxide dismutase (sod) and sirtuin 1 (sirt1) genes and superoxide dismutase enzyme (SOD) and sirtuin 1(SIRT1) protein activity assays were conducted at 1 mg/mL of CIFE. After administrating CIFE, the lifespan of the yeast was significantly prolonged in comparison with the untreated group (P<0.05) and also increases the yeast cell viability under oxidative stress by 4 mM H₂O₂ by spot assay. The Phloxine B stain method also further proves yeast cells' viability increased under oxidative stress by CIFE treatment. The reactive oxygen species (ROS) assay revealed that CIFE notably reduces the oxidative stress in treated yeast cells (P<0.05) comparing to untreated yeast cells.
- PublicationCharacterization Of Astaxanthinrich Xanthophyllomyces Dendrorhous Extract From A Hyperproducing Mutant And Its Effects On Breast Cancer Cells(2022-12)Khaw, Shin YuanAstaxanthin is a carotenoid with multiple health benefits including antioxidant and anticancer properties. An astaxanthin-hyperproducing X. dendrorhous mutant was generated due to the growing preference for natural pigment. This work aimed to study the astaxanthin-rich X. dendrorhous extracts and its effect on MCF-7 and MDA-MB- 231 cells. Spectrometric, TLC and HPLC analysis evidenced that mutant M34 produced astaxanthin in higher quantity and purity than the wild type. Mutant astaxanthin extract showed a higher scavenging activity compared to wild type astaxanthin extract in DPPH assay. MTT assays proved that wild type and M34 astaxanthin extracts were not toxic towards the non-cancer origin MCF-10A cells. Wild type and mutant astaxanthin extracts exhibit growth-inhibitory effect in dosedependent manner in MCF-7 and MDA-MB-231 cells. Mutant astaxanthin extract showed a lower IC50 than wild type extract and was more effective in inhibiting both MCF-7 and MDA-MB-231 cells. The IC50 values of wild type and mutant astaxanthin extracts did not exceed the required upper limit of 30 μg/ml.
- PublicationDeciphering S-Phase Protein Kinase 2 (Skp2) Mediated Regulation Of Telomerase Reverse Transcriptase (Tert) In Kasumi-1, T (8;21) Acute Myeloid Leukemia (Aml) Cell Line(2023-06)Rosli, Nur Aliaa ArinaAcute myeloid leukemia (AML) is the most common acute leukemia in adults and is characterized by immature myeloid cell proliferation. S-Phase Protein Kinase 2 (SKP2) is a cell cycle regulator and shown to be overexpressed in AML patients. SKP2 potentiates to cause cell proliferation and division. Studies shown prolonged SKP2 knockdown suppress telomerase reverse transcriptase (TERT) expression in t(8;21) AML cells in vitro. Nevertheless, the molecular mechanism of TERT regulation by SKP2 remain unclear. Therefore, the aim of this study was to investigate the TERT mechanism by SKP2 in AML.SKP2 was suppressed in Kasumi-1 and THP-1 via siRNA mediated gene knockdown. In this study, TERT expression reduced at gene and protein level after SKP2 suppression in non t(8;21) AML cells. Accordingly, telomerase activity was also reduced in non t(8;21) AML cells. Result obtained show that c-Myc and FOXO3 did not play a direct role in SKP2 mediated TERT regulation in t(8;21) and non-t(8;21) AML cells at gene level. However, different pattern in c-Myc protein expression was observed in non t(8;21) AML cells. Another observation were made in t(8;21) AML cells after AML1/ETO knockdown where c-Myc was up-regulated at gene and protein level. Due to increase in c-Myc expression levels, chromatin immunoprecipitation (ChIP) was carried out and increased observed binding of c-Myc to the TERT promoter was observed after AML1/ETO down-regulation. However, c-Myc binding to the TERT promoter failed to induce TERT transcription in t(8;21) AML cells.Other proteins related to TERT mechanism xxii observed were Rb, E2F1 and CDKN1B. Hypophosphorylation of Rb was observed up-regulated in non t(8;21) AML cells after SKP2 knockdown yet no significant difference in E2F1 protein expression was observed. Accumulation of CDKN1B was markedly related with suppression of SKP2 in this studies.
- PublicationDevelopment Of A Flexible Stable Mammalian Antibody Expression Pipeline(2023-02)Jacqueline Mark Kar KeiMammalian antibodies are promising tools for biopharmaceutical use because of their high specificity and low toxicity. These proteins are favorable for both therapeutic and diagnostic purposes and its ever-growing demand calls for high productivity cell lines. Mammalian cell lines can express antibodies transiently or stably. Although the transient expression system has the benefits of rapid production, it is more suitable for short-term protein production and the initial phase of antibody testing. Meanwhile, a stable expression system is the go-to for long-term mass protein production because of its ability to express proteins in a reproducible, scalable, and reliable manner with no batch-to-batch variation.
- PublicationElucidation Of Erk1/2/C-Myc/P53 Signaling Pathways Involved In Andrographolide-Induced Antiproliferative Activity In Human Glioblastoma Dbtrg-05mg Cell Line(2023-03)Nurul Syamimi Binti OthmanThe whole study evaluates andrographolide's anticancer effectiveness and potential molecular pathways using a glioblastoma multiforme (GBM) cell line.
- PublicationInhibitory Effects Of Andrographolide In Pc-3 Cell Line And The Induction Of Apoptosis Via The Involvement Of Caspases 3, 8 And 9(2023-04)Manimaran, JananyAndrographolide is a labdane diterpenoid isolated from the plant Andrographis paniculata. This substance has numerous medicinal uses, notably anticancer effects. A previous study has revealed that andrographolide inhibits the growth of lung, brain, colon, and breast cancer cells. Due to a lack of research, the knowledge of andrographolide's anti-cancer effects on prostate cancer cells is relatively poor. In the current study, andrographolide was assessed on PC-3 cells, an aggressive androgen-independent prostate cancer cell line. Cytotoxicity analysis is vital in drug discovery research for assessing the biocompatibility of the drug being used on cancer cells. This work used the WST-1 assay to determine the cell survival of PC-3 cancer cells and Hs27 normal cells exposed to varying doses of andrographolide (0-200 μM). The results indicate that andrographolide dose-dependently suppresses the viability of PC-3 cells but not Hs27 cells. The NCI considers the LC50 value of 26.42 μM (after 48 hours of incubation) is acceptable. For the first time in this study, three different concentrations of andrographolide were used: control, half LC50, and LC50 (0, 13.21, and 26.42 μM) in all subsequent analyses. Metastasis is essential for the disease to progress; hence, the scratch assay and the transwell invasion assay were used to test andrographolide on PC-3 cells.
- PublicationModifications Of Nk-92mi Cell Line Through Third Generation Lentiviral Vector(2023-01)Chin Ding ShengIn this study, NK-92MI cell line was transduced with RRL lentivirus carrying the genes for either intracellular GFP or surface CD16 receptor, with the addition of polybrene polycation. Results showed that GFP expression averaged at 3.49 % without polybrene and averaged at 7.18 % when polybrene was added. Extracellular CD16 expression averaged at 1.03 % without polybrene and increased to an average of 3.12 % with polybrene. This study demonstrated the feasibility of using the third generation RRL lentiviral vector for NK cell transduction.
- PublicationRole Of Polysaccharides From Pleurotus Sajor-caju In The Immunomodulation Of Thp-1 Human Macrophage Cells(2022-06)Mokhtar, MunirahMedicinal mushrooms have a diverse range of biological effects, which are excellent sources for pharmaceutical and nutraceutical use. They possess various valuable medicinal properties, including antitumor, hypocholesterolemic, anti-atherogenic, and anti-oxidative properties. Pleurotus sajor-caju is one of the Pleurotus spp. belonging to phylum Basidiomycota, well-known as oyster shaped mushroom (basidiocarps) and generally called oyster mushroom (OM). The present study was aimed to determine the ability of the water-soluble polysaccharides extract from Pleurotus sajor-caju extracts (PSC) in inducing the proliferation, immunostimulating and anti-inflammatory effects on THP-1 macrophage cells. First, the total carbohydrate and β-glucan contents were determined using the individual assay test. Next, the cell proliferation response was evaluated using the MTS assay. To understand the immunostimulating effect of PSC extract at the gene level, the expressions of selected cytokine genes in THP-1 cells were evaluated using real-time PCR. Nitric Oxide (NO) production and inducible nitric oxide synthase (iNOS) expression level were measured in the LPS-stimulated macrophages. The results obtained showed that 95% of total carbohydrate and 89% of β-glucan contents were detected in 50 mg PSC extracts, respectively. PSC extract was found to induce a marked increase in proliferative response on macrophage THP-1 cell in a dose-dependent manner, ranging from 10 μg/ml up to 80 μg/ml. At transcription level, PSC stimulated the expression of tumor necrosis factor-alpha (TNF-α), interleukin 1β (IL-1β), and interleukin 6 (IL-6) significantly (p<0.05) at various dosages of PSC extract.
- PublicationSynergistic Activity Of Ceftriaxone And Polyalthia Longifolia Leaves Extract Against Methicillin Resistant Staphylococcus Aureus (Mrsa)(2021-09)Ranjutha A/P ValiappanMethicillin‐resistant Staphylococcus aureus (MRSA) infection has become one of the most significant pathogenic bacterial infection health issues in developing countries. Furthermore, the most important mecA gene that is encodes a novel penicillin-binding protein PBP2a contributes to the methicillin resistance in MRSA strains. Therefore, new alternative strategies are needed to address this issue by developing a new antimicrobial agent, modifying the existing antibiotic activity with a combination of plant extracts as resistance modifying agents, or using the plant extract combined with existing antibiotics against resistant bacteria. Consequently, the current study was conducted to evaluate the antibiotic modifying activity and synergistic effects of methanol extract Polyalthia longifolia (Sonn.) Thwaites (Annonaceae family) against MRSA in combination with ceftriaxone. The antibiotic modifying activity and synergistic effects of P. longifolia leaf methanol extract (PLLME) were evaluated in this study by disc diffusion method, broth microdilution method, checkerboard microdilution method, and detection of the mecA gene suppression by multiplex Polymerase chain reaction (PCR).