Institut Penyelidikan Perubatan Molekul - Tesis

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Antiaging Activity Of Polyphenol Rich Calophyllum Inophyllum Fruit Extract In Aging Mutants Of S. Cerevisiae
(2022-12)
S. Venugopal, Kavilasha
Calophyllum 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.
Assessment Of Antitumor Activity Of Standardized Polyalthia Longifolia Leaf Extract In Hela Cell Tumor Xenograft Mouse Model
(2022-08)
Braganza, Cilwyn Shalitha
P. longifolia is a traditional medicinal plant with a rich source of biologically active phytochemicals. Investigations on methanolic leaf extract were reported to exert potent in vitro anticancer effects on HeLa human cervical cancer cells. Current treatment strategies for cervical cancer are facing challenges such as increased drug toxicity, chemoresistance, and limited treatment options, thus making it imperative for the discovery of safe and effective green anticancer agents. Though P. longifolia is a promising anticancer plant, the in vitro findings by themselves are insufficient to translate to human use and therefore require in vivo preclinical testing. Currently, there are no reports on further investigation of P. longifolia leaf extracts on in vivo animal tumor models, which limits the clinical utility of this plant. Hence this study was conducted to evaluate the in vivo antitumor effects of P. longifolia methanolic leaf extract on HeLa cells xenografted tumors developed in athymic nude mice.
Bzd9l1: Elucidation Of Its Anti-angiogenic Potential In In-vitro And In-vivo Colorectal Cancer Models
(2023-09)
Subramaniam, Ayappa V.
Colorectal cancer (CRC) is the third most common cancer globally. CRC depends largely on angiogenesis for growth and metastasis. Much effort has been made to selectively target the angiogenic pathways to restrain tumour growth. However, some CRC patients become resilient to these anti-angiogenic drugs and standard therapies. The class III histone deacetylase family of sirtuins (SIRTs) has been closely linked to cancer progression but less is known about its activity in regulating tumour angiogenesis. BZD9L1 is a novel sirtuin inhibitor with demonstrated anti-cancer activities. This study aimed to investigate the anti-angiogenic potential of BZD9L1 on EA.hy926 endothelial cells (EC) in vitro and HCT116 tumour xenograft nude mice. The in vitro experiments comprised of cell viability assay, scratch wound assay, tube formation assay, spheroid sprouting assay, western blotting, angiogenesis array, cell cycle and apoptosis analysis via flow cytometry and finally, indirect co-culture model. Nude mice tumour xenograft model was used for the in vivo study, where hematoxylin and eosin staining was done to study the percentage of necrosis in the tumour section and immunohistochemistry was conducted to investigate the protein expression of Ki67 and CD34. BZD9L1 was shown to reduce cell viability, cell migration, tube formation, and spheroid sprouting of EC. BZD9L1 at 10μM was also shown to inhibit SIRT2 and SIRT3 protein in EA.hy926 cells. Angiogenesis array results revealed that the compound reduces the cytokine concentration of Angiogenin, bFGF, PDGF-BB, and PIGF significantly (P * < 0.05) compared to the control group
Characterization Of Astaxanthinrich Xanthophyllomyces Dendrorhous Extract From A Hyperproducing Mutant And Its Effects On Breast Cancer Cells
(2022-12)
Khaw, Shin Yuan
Astaxanthin 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.
Chemopreventive Effects And Proteomic Analysis Of Myo Inositol Treatment On Human Prostate Cancer Cell Line (Du-145)
(2022-02)
Islam, Mohammad Jahidul
Prostate cancer remains the second most frequent diagnosed cancer in men and is the third leading cause of cancer related death, despite many available treatment options. The inefficiency of existing therapies and their adverse effects have led scientists to seek new treatments for this disease. The identification and development of new anti-proliferative agents for prostate cancer is a major innovation in treating this deadly disease. Although the synthetic compound Myo-inositol is widely used in cancer research, the actual anti-cancer function of these chemicals is still not well understood. This study was therefore intended to unveil the chemopreventive effects of Myo-inositol on prostate cancer cells (DU-145). The growth inhibitory effect and the IC50 value were demonstrated by Myo-inositol at 0.06 mg/ml (**p<0.05). It is suspected that cell deaths are related to apoptosis induction and cell-cycle arrest. Treatment with Myo-inositol has been further identified by induction of early and late apoptosis (***p<0.01). Apoptosis can also be detected using DNA fragmentation and Hoechst 33342 fluorescent dye stain analysis. Myo-inositol caused an alteration to the cell cycle regulation on DU-145 cell line at G0/G1 and S phase, respectively (***p<0.01). Protein identification results via 2D electrophoresis and LC-MS/MS analysis showed 19 and 23 proteins identified in control and Myo-inositol treated samples, respectively.
Deciphering 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 Arina
Acute 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.
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.
Development And Characterization Of Monoclonal Antibodies Against Ancylostoma Caninum Ancylostoma-Secreted Protein 5 (Asp5) By Phage Display
(2024-07)
Brenda Song, Pei Chui
Parasitic nematodes such as Ancylostoma caninum engages in a complex life cycle, in which it can impact the hosts at its various stages of development and infection. During infective stage, Ancylostoma caninum releases a multitude of proteins to modulate the host’s immune response, which ensuring its survival within the host environment. Among these proteins, Ancylostoma-secreted protein 5 (ASP5) is critically involved in the parasite-host interaction such as the immunomodulation, tissue invasion and facilitating the blood feeding process, hence making it a promising target for intervention strategies aimed at controlling hookworm infections among canines. By harnessing phage display technology, this study aims to develop monoclonal antibodies targeting Ancylostoma-secreted protein 5 (ASP5), by employing the human naïve scFv antibody library through in vitro biopanning. The library was subjected to a total of three rounds of panning against 10 μg of ASP5 antigen, with progressively stringent washing conditions (10, 20 and 30 washes respectively) to isolate the clones. This study delves into Ancylostoma-secreted protein 5 as an antigenic candidate for monoclonal antibody development in which it was expressed using the bacterial strain of BL21(DE3) and purified with Immobilized Metal Affinity Chromatography (IMAC) purification method using the nickel-charged affinity resin. The developed antibody was characterized on its antibody-antigen interactions, including cross-reactivity and binding strength via Enzyme-linked immunosorbent assay (ELISA).
Development Of A Chronological Life Span Assay For Ageing Studies In Saccharotnyces Cerevisiae
(2019-06)
Ong, Tee Gee
The Saccharomyces cerevisiae. chronological life span (CLS) is the length of time that yeast cells survive in a non-dividing state. In the laboratory, CLS is typically measured by its optical density at the post-diauxic phase that begins approximately two days after initial inoculation. In this study, the development of a CLS assay on a 96-well microplate with different yeast strains and growth parameters (media and time) were carried out. After the evaluation of four different strains, yeast strain BY25929 was found to be the most suitable in tenns of growth for the CLS assay. A total of 24 ageing-related proteins were selected for assessment in the CLS assay.
Development Of A Flexible Stable Mammalian Antibody Expression Pipeline
(2023-02)
Jacqueline Mark Kar Kei
Mammalian 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.
Development Of Leishmania Tarentolae Expression Systems For Different Recombinant Antibody Formats
(2024-04)
Jing Yi, Lai
Production of recombinant antibodies has been an important topic for biomedical applications. Currently, expression of monoclonal antibodies is mainly performed using mammalian cell lines such as Chinese hamster ovary (CHO) cells and Human embryonic kidney 293 (HEK293) cells due the advantage of human-like post-translational modifications. However, the use of mammalian cell lines has some drawbacks especially in term of cost. The introduction of Leishmania tarentolae as an expression host is viewed as an interesting alternative due to their post-translational modification, which is similar to human but is more cost-effective for expression and maintenance. In this study, the use of L. tarentolae was explored as a platform to express recombinant antibodies in scFv-Fc and IgG format. A single chain fragment variable (scFv) clone against long-chain neurotoxin (LNTX) of Naja kaouthia was identified from naïve human antibody library. The clone was expressed and characterized in Escherichia coli as scFv and further converted to scFv-Fc and IgG format for expression in L. tarentolae. The expression of IgG was achieved using a bicistronic vector system. A total of two constructs, encompassing both possible position of the heavy chain (HC) and light chain (LC), were expressed. The construct with the HC-LC gene arrangement was found to perform better in term of yield. The stability of the IgG at 26ºC and 45ºC was also examined. While the expression performance of antibody clone is dependent on the antibody gene sequence, the study shows the potential application of L. tarentolae for expressing both scFv-Fc and IgG. In conclusion, the L. tarentolae expression system can be touted as a possible alternative system for the expression of scFv-Fc and IgG antibody clones.
Development Of Novel And Environmentally Friendly Bio- Active Dermal Drug Delivery Wound Patch For Wound Healing
(2024-09)
Sunder, Sharminy Pirem
Having a wound-healing dermal patch or medicated dressings that work well in wound healing and doesn't harm the environment is crucial. The commercially available synthetic dermal patches, plasters, and wound dressings in the market today are non- biodegradable, posing significant environmental issues. As a result, it is a great idea to reuse agro-waste to make a biodegradable dermal wound patch and, eventually, reduce environmental problems. Elaeis guineensis plant was reported with good wound healing activity in the literature and is commonly left as agro-waste at plantation areas. Therefore, this study was conducted to develop an environmentally friendly bio-active dermal drug-delivery wound patch containing E. guineensis for wound healing. The E. guineensis leaf was extracted using methanol by maceration extraction techniques, which yielded a dark brown paste-like crude extract. Subsequently, E. guineensis leaf extract was blended with polyvinyl alcohol (PVA) to develop a biodegradable dermal wound patch. Several physicochemical properties were evaluated and reported, such as appearance, average weight, thickness, moisture content, viscosity, and pH. The developed dermal wound patch had an average thickness of 0.020 mm and a weight of 3.87 mg with 17.61% of moisture content. The pH of the prepared dermal wound patch solution was 6.97, with a viscosity value of 2.56 cp. These results showed that the developed dermal wound patch is an appropriate and safe candidate to be utilized as a wound-healing agent.
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.
Elucidation 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 Othman
The whole study evaluates andrographolide's anticancer effectiveness and potential molecular pathways using a glioblastoma multiforme (GBM) cell line.
Evaluation Of Toxicity, Pharmacokinetic And Clastogenicity Profile Of Ethyl 2-[4[(Piperidin-1-yl) Phenyl]-1h-benzimidazole-5-carboxylate (Bzd9l1), Novel Sirtuin Inhibitor
(2022-05)
Lee, Yeuan Ting
The mammalian sirtuins (SIRTs) have been linked to various diseases including cancer, diabetes, neurodegenerative and cardiovascular diseases. Thus, SIRTs are attractive targets for the development of pharmaceuticals. The lack of potential SIRT inhibitors in cancer clinical trials highlights the need to develop a potent SIRT modulator as an anti-cancer therapeutic. Studies in the lab have highlighted the capability of a lead sirtuin inhibitor (BZD9L1) to reduce colorectal tumour growth in vitro and in vivo by modulating different cancer pathways in colorectal cancer with different mutation profiles. Also, synergistic effects of BZD9L1 in in vitro assays and tumour xenograft study when used in adjunct with 5- Fluorouracil (5-FU) further support the promising therapeutic effects of BZD9L1 as anticancer regime. Nevertheless, the toxicology profile of this compound remains unknown. To our knowledge, no prior work has reported the regulation of SIRTs in the liver and kidney by a small molecule inhibitor in a drug toxicity study. Therefore, this project aims to determine the toxicology, molecular regulation of toxicity, pharmacokinetics and clastogenicity profiles of BZD9L1 in in vitro or in vivo models.
Exploring The Immunomodulation Profiles Of The Thp-1 Human Macrophage-derived Cell Line Mediated By Shigella Flexneri
(2022-06)
Shabani, Nor Raihan Mohammad
Shigellosis is the primary cause of severe diarrhea, especially among children less than five years old. The occurrence of multi-drug resistant strains of Shigella species has contributed to the ineffectiveness of the existing treatment. Vaccination has become an essential requirement for preventing shigellosis. The invention of an epitope-based vaccine has directed the development of a new vaccine design model. Macrophages are specialized APCs that process antigens and then present the processed antigens to T-cells through HLA molecules. The immunomodulatory profiles of the macrophages infected by the clinical strains of S. flexneri 2a have remained undefined. THP-1-derived macrophages, as the model of human macrophages, were infected independently with mild and virulent strains of S. flexneri 2a at standard growth conditions. The gene expression level of inflammatory mediators was determined using qPCR, while NO production was measured using a commercial NO assay kit. The significant up-regulation of TNFα, IL-1β, IL-6, IL-12, iNOS, and NO indicates the pro-inflammatory reaction to S. flexneri 2a infection. The virulent strain also markedly increased the expression levels of the anti-inflammatory cytokine mRNAs. HLA class II gene expression was also investigated to extend the immunopeptidomics study and found the significant up-regulation of HLA class II by the Shigella-infected macrophages. LC-MS/MS-based immunopeptidomics strategy was employed to identify the peptides that can be used as potential candidates for the epitope-based Shigella vaccine development
Genomic Expression Profile Of Human Papillomavirus 16 And 18 – Associated Pre-cancerous Lesions And Cancer Of The Cervix
(2022-06)
Balasubramaniam, Shandra Devi
Cervical cancer is one of the most common cancers in women and is caused by high-risk human papillomavirus (HPV) infection. The integration of HPV into host cervical epithelial cells causes genetic alterations and consequent changes in gene expression affecting downstream molecular pathways, leading to the development of cervical cancer. This study aimed to profile the genomic signatures involved in the pathogenesis of HPV 16 and 18 associated pre-cancerous lesions (cervical intraepithelial neoplasia, CIN) and squamous cell carcinoma (SCC) of the cervix, in comparison to the normal cervix. The differential mRNA expression profiles were determined, to evaluate the expression of up-regulated and down-regulated host genes and to evaluate the perturbed molecular pathways involved in cervical carcinogenesis. In this study, 29 formalin-fixed paraffin-embedded (FFPE) tissues including low-grade CIN (LGCIN), high-grade CIN (HGCIN), SCC, and normal cervix were screened for HPV 16 and HPV 18 using immunohistochemistry and qRT-PCR method. Of these, 9 HPV-positive (3 LGCIN, 3 HGCIN, 3 SCC) and 3 normal cervix (HPV-negative) tissue samples were microdissected to obtain regions of interest in the squamous epithelium before RNA extraction. Transcriptomic profiling was performed using the Affymetrix, GeneChip Human Transcriptome Array 2.0 (HTA 2.0) and the nCounter® PanCancer Pathway Array, Nanostring to identify differentially expressed genes (DEGs) and significantly associated pathways in each stage of cervical cancer development.
Identification Of Antimalarial Compounds And Their Mode Of Actions
(2022-10)
Mahmud, Fauze
Malaria is one of the most significant infectious diseases in the tropics, claiming around half-million lives annually, mainly due to Plasmodium falciparum (P. falciparum) infections. To date, P. falciparum has developed full or partial resistance towards all three classes of antimalarials, including against the artemisinin. Hence, it may jeopardize the efficacy of the current antimalarial treatment regimen, the Artemisinin Combination Therapy (ACT), in the near future. Therefore, there is an utmost need to identify a new antimalarial compound with a novel mode of action (MoA). This study mainly focused on identifying antimalarial compounds from soil microorganisms isolated from Malaysia and Japan. The crude extracts of these strains showed potent antimalarial activity in a preliminary assay against Pf 3D7 (wild type). The crude extracts of Malaysian strains also inhibited the activity of human GSK-3β (75 % similar to P. Falciparum GSK-3 (Pf GSK-3)).
Identification Of Immunogenic Proteins From Cysts And Trophozoites Of Giardia Lamblia By Immunoassay
(2024-07)
Roshidi, Norhamizah
Giardia lamblia exists in two stages in the life cycle; the motile trophozoite and the resistant cyst that is highly adapted to harsh environments, infecting humans and animals with a disease known as giardiasis. Giardiasis is a significant public health concern, with an estimated 280 million symptomatic human infections occurring each year and contributing to 1.6 million cases of diarrheal death in 2016. In Malaysia, cases of giardiasis reportedly ranged from 2.6% to 25% from 1970 to 2000 and rise to 10.4% to 28.3% from 2002 to 2019. However, studies on giardiasis are very limited and mostly focused on the variation of gene expression and vaccine development. Therefore, this study was conducted to characterize Giardia proteins that may lead to a better understanding of Giardia-immunogenic proteins potential for diagnostics and therapeutic applications. The approach of this study involves in-vitro cultivation of G. lamblia trophozoites and cyst in different culture medium, protein extraction and separation using OFFGEL™ as an improved pre-fractionation step prior to immunoassays and mass-spectrometry analysis to identify the identity of potential immunogenic proteins. The maximum number of trophozoites of 9.6 x 106 cell/ml was successfully obtained in the TYI-S-33 medium at pH 7 for 72 hours at 37°C, while the maximum number of cysts of 7.45 x 105 cell/ml was produced in complete modified TYI-S-33 medium containing 0.25 mg/ml of porcine bile at pH 7.8 for 72 hours at 37°C.

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