Institut Penyelidikan Perubatan Molekul - Tesis

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In Vitro And In Vivo Effects Of Bisphenol A On Cell Growth, Lipid Accumulation And Inflammation In Liver Cell Line, Rat Adipose And Colorectal Tissues
(2024-03)
Ismael, Ismael Layla Qasim
Bisphenol A (BPA) is an exogenous endocrine disruptor that mimics hormones closely associated with health complications, such as cancer progression. BPA is also linked to an increase in the prevalence of obesity and obesity-related diseases due to its obesogenic action. The present study aimed to evaluate the effects of BPA on human liver cells and animal tissues. The BPA-treated THLE-2 cell viability was first evaluated using an MTT assay. Cytoplasmic lipid accumulation was then examined in the BPA-treated THLE-2 cells using Oil Red O staining. The expression of various adipogenesis genes in lipid metabolism, including clusterin, PPARs, CYPs, UCP1, and BRS3 in BPA-treated THLE‑2 cells, was investigated using real-time PCR. Subsequently, flow cytometry measured the DNA synthesis in BPA-treated THLE-2 cells was performed using BrdU/PI staining. The pro-inflammatory proteins IL6 and CCL2 in the conditioned media and cytoplasmic proteins of the BPA-treated THLE-2 cells were examined by ELISA. After that, various adipogenesis genes’ expression in lipid metabolism, including clusterin, PPARs, CYPs, UCP1 and BRS3, in adipose and colorectal tissues of the rats fed a diet (normal or high fat) supplemented with and without BPA was investigated using real-time PCR.
The Antiaging Activity Of Polyalthia Longifolia (Sonn.) Thwaites Methanolic Extracts On Saccharomyces Cerevisiae By611 Yeast Cells
(2024-03)
Manisekaran, Hemagirri
The pursuit of natural-based antiaging agents has gained momentum in recent years. Polyalthia longifolia, a valuable indigenous medicinal plant, is widely used in traditional medicines due to its high polyphenol concentration. This study assessed the anti-aging potential of P. longifolia leaf methanolic extracts (PLME) using Saccharomyces cerevisiae strain BY611 as a model organism. In this research, an integrated approach, encompassing in vitro, in situ assays and RNA sequencing techniques was employed to investigate the anti-aging properties of PLME. The methanolic extract was prepared, and rutin quantification yielded 0.013 μg/mL (1.30%) rutin content in 1000 μg of PLME. Furthermore, an optimal concentration of 1 mg/mL PLME was determined for achieving maximum anti-aging effects. Intriguingly, PLME treatment significantly (p < 0.05) extended both replicative and chronological lifespans and total lifespan while enhancing cellular proliferative potential compared to the vehicle control group. Corroborated by light microscopy, PLME also promoted longevity in BY611 yeast cells by mitigating the progression of large apoptotic-like Type 3 vacuoles.
Development Of Tcr-like Single Domain Antibody By Incorporating Immunoglobulin Isotypes For Latent Tuberculosis Diagnostics And Therapeutics Application Against Heat Shock Protein 16 Kda Peptide Presented By Hla-a2
(2024-08)
Liu, Hua Qiang
Heat shock protein 16 kDa (HSP 16 kDa) is essential for the survival of latent Mycobacterium tuberculosis. Therefore, a peptide-MHC presentation of HSP 16 kDa could be a potential diagnostic and therapeutic target for latent tuberculosis (LTB). This study aimed to generate different immunoglobulin isotype pairings on a TCR-like single-domain antibody (sDAb) and subsequently investigate its diagnostic and therapeutic potential in LTB, utilizing a model cell presenting the target peptide. A previously generated TCR-like sDAb that can bind to HSP 16 kDa was first fused to a human IgG1, IgG2, IgG4, IgA, IgD, IgE, and IgM Fc-receptor via a linker. The fusion product, sDAb-Fc, was expressed in HEK293-F cells and subsequently purified. Its diagnostic potential was investigated via cell-based ELISA utilizing MCF-7 cells peptide-pulsed with HSP 16 kDa peptides. Antibody-dependent cell-mediated cytotoxicity (ADCC) of MCF-7 cells was also conducted to investigate its therapeutic potential. Finally, TCR-like sDAb-IgG1, IgG2, and IgG4 were successfully produced transiently in HEK293 F cells and purified using protein A chromatography.
Elucidating The In Vitro Antiproliferative Properties And Associated Mechanisms Of Momordica Cochinchinensis Spreng (Gac Fruit) Aqueous Extract Using Colorectal Cancer Origin Cell Lines
(2024-07)
Thavamany, Priscilla Jayanthi
Momordica cochinchinensis S., also known as gac fruit, is a tropical fruit native to Southeast Asia. However, it has not yet been fully explored in Malaysia. The extracts obtained were aril water extract (AW), pulp water extract (PW) and seed extracts (SW), and its ethanolic counterpart, namely aril extract (AE), pulp extract (PE) and seed extract (SE). The focus of this study in terms of their phytochemical composition, antioxidant, antimicrobial, antiproliferative, and wound healing properties. Both water and ethanolic extracts of the aril, pulp and seed contain alkaloids, flavonoids, saponins, volatile oil and reducing sugars. However, glycosides were only present in water extracts (AW, PW, SW), meanwhile tannins were detected only in SW. The PW exhibited an increased level of total phenolic content (TPC); 0.0215 ± 0.00060 mg GAE/g whereas, total flavonoid content (TFC) was quantitated at 0.083 ± 0.022 mg QE/g FW (TFC), respectively. Apart from that, the PW extract also exhibited potent antibacterial activity, with MIC values between 5 and 20 mg/ml and MBC values between 10 and 20 mg/ml against E. coli, P. aeruginosa, S. flexneri, and B. cereus. The concentrations ranged between 1μg/ml and 10 μg/ml of PE and SW extracts showed positive effects in the wound healing experiment. The aril of the fruit is known for its high concentration of carotenoids, such as beta-carotene and lycopene. These compounds are powerful antioxidants and have been shown to have an anti-proliferative effect on various types of cancer-causing cells.
Development And Characterisation Of Nutraceutical Herbal Tea From Polyalthia Longifolia Leaves
(2024-01)
Alkatib, Huda Hisham Sultan
The development of nutraceutical herbal tea from native medicinal plants has gained popularity due to its numerous health benefits. Polyalthia longifolia is well documented for its valuable ethnomedicinal properties, but no commercial products are available based on this plant. Therefore, the new MyAyush® nutraceutical tea was developed in this study from P. longifolia leaves. Initially, to establish the standards for the P. longifolia leaves, which can be used to correctly identify the plant in the future and ensure the quality of MyAyush® tea, the evaluation of proximate, phytochemical, FTIR and heavy metals analyses were conducted. Subsequently, the sensory evaluation, in vitro antioxidant, anticancer, and antitumorigenic activities of the MyAyush® tea were also assessed. The proximate, phytochemical, and heavy metals analyses revealed the presence of crucial proximate composition (6.9% ash, 25% crude fibre, 4.5% crude fat, 18.9% crude protein, 6.6% moisture, and 63.1% carbohydrates), 32 phytochemicals, as well as the presence of, mercury, cadmium, lead and arsenic in low quantities and within the permissible limit. The standardization of MyAyush® tea was performed by quantifying the rutin chemical marker, which was found in a concentration of 1.2 μg/mL (0.12%). Besides, the FTIR analysis showed the presence of ten peaks' areas in the range of 3500 to 400 cm-1 for various specific functional groups. The sensory evaluation results indicated that the average general acceptance level of MyAyush® tea (6.6) was close to the acceptance level (7) of the commercially available tea.

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