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Prevalence of non-alcoholic fatty liver disease (NAFLD) is increasing steadily every year affecting all population both Western and Asian countries. The current treatments available for NAFLD are non-conclusive warranting newer effective pharmacological agents. Newly formulated agents require prior testing using animal models. However, in developing countries, these models are often costly. The possibility of using more affordable animal model in local settings should be investigated. In this study, ten Institute of Cancer Research (ICR) and seven B6.Cg-LepOb/J leptin-knockout (JAX) male mice were recruited. Five ICR and all JAX mice were subjected to high-fat diet (60% kcal fat) and remaining ICR mice were given standard diet (SD) for six weeks. Body weight and food intake were measured weekly while abdominal circumference, random blood glucose and liver span were measured at the end of the HFD study. Livers collected were subjected to histology assessment. Compared to ICR group, JAX group presented with significantly higher body weight (58 ± 0.72, p

The notable ability of human liver cytochrome P450 3A4 (CYP3A4) to metabolize diverse xenobiotics encourages researchers to explore in-depth the mechanism of enzyme action. Numerous CYP3A4 protein crystal structures have been deposited in protein data bank (PDB) and are majorly used in molecular docking analysis. The quality of the molecular docking results depends on the three-dimensional CYP3A4 protein crystal structures from the PDB. Present review endeavors to provide a brief outline of some technical parameters of CYP3A4 PDB entries as valuable information for molecular docking research. PDB entries between 22 April 2004 and 2 June 2021 were compiled and the active sites were thoroughly observed. The present review identified 76 deposited PDB entries and described basic information that includes CYP3A4 from human genetic, Escherichia coli (E. coli) use for protein expression, crystal structure obtained from X-ray diffraction method, taxonomy ID 9606, Uniprot ID P08684, ligand–protein structure description, co-crystal ligand, protein site deposit and resolution ranges between 1.7[Formula: see text]Å and 2.95[Formula: see text]Å. The observation of protein–ligand interactions showed the various residues on the active site depending on the ligand. The residues Ala305, Ser119, Ala370, Phe304, Phe108, Phe213 and Phe215 have been found to frequently interact with ligands from CYP3A4 PDB. Literature surveys of 17 co-crystal ligands reveal multiple mechanisms that include competitive inhibition, noncompetitive inhibition, mixed-mode inhibition, mechanism-based inhibition, substrate with metabolite, inducer, or combination modes of action. This overview may help researchers choose a trustworthy CYP3A4 protein structure from the PDB database to apply the protein in molecular docking analysis for drug discovery.

Drug-drug interactions (DDI) are the most common cases that occur in our healthcare in which are very alarming as it may lead to severe complications. Consumption of natural products concomitantly with conventional drugs or treatment using polypharmacy have become the norm that promoting the potential of pharmacokinetic or pharmacodynamic drug interactions as the combination may mimic, increase or reduce the effects of the drug or the herb which could result in clinically significant interactions. CYP2C9 is the second major isoform from CYP450 family of enzyme, which responsible in phase 1 metabolism of 15-20% clinical drugs. Up to date, many substrates of CYP2C9 have been discovered and these discoveries may open more doors for potential drug-drug interactions in patients. Many studies have been done to evaluate the effect of drugs on the activity of CYP2C9 and how it influenced the effectiveness of therapy in patients. Various data regarding CYP2C9 related DDI from in vitro, in vivo and clinical studies were critically discussed in this review to provide insights on how these drugs and natural products may exhibit drug interactions clinically. This review could be beneficial reference material for health practitioners and researchers.

Aims: This study aims to develop an efficient, and inexpensive transfection procedure using low cost polyethylenimine (PEI) as transfection reagent for overexpression of CYP2C9 in WRL 68 host cells. Introduction: Overexpression of CYP450 isozymes such as CYP2C9 has been a reliable approach in developing a high specificity in vitro tool for inhibition study of this liver enzyme. Transient or stable transfection are used to insert plasmid DNA into a mammalian host cell in order to produce proteins. There are many efficient transfection reagents available in the market for this purpose, however, it can be expensive, and a low-cost alternative is favorable. Methodology: Cytotoxicity of PEI was screened on four cell lines namely WRL 68, HepG2, MCF-7 and A549 using MTS assay. WRL 68 was transiently transfected with CYP2C9 plasmid using PEI and DNA concentration as well as transfection time was optimized for the best efficiency and expression. Expression of CYP2C9 protein was measured using qPCR and further evaluated with western blot analysis. Results: IC50 values of PEI are 0.327 ± 0.013 mg/mL (WRL-68), 0.395 ± 0.037 mg/mL (HepG2), 1.159 ± 0.032 mg/mL (A549) and 1.281 ± 0.000 mg/mL (MCF-7). Combination of 3ug plasmid DNA with 2.8 μM of PEI and 10.5 mM NaCl resulted in the highest transfection efficiency and expression after 48 hours. Conclusion: A low-cost and efficient PEI transient transfection procedure was optimized for CYP2C9 overexpression and useful for the purpose of further development of cell-based enzyme inhibition model.

Christia vespertilionis locally known as ‘rerama’ has gained much attention as herbal preparation to treat various ailments and diseases although there is no conclusive evidence of its therapeutic values. Self-medication is commonly practiced by patients, underpinned by health believes that affect their adherence to medication regimens, and impacting on treatment outcomes. Therefore, in this study the phytochemical profile and potential interaction of this plant extracts with conventional chemotherapy drug cyclophosphamide were investigated. Water decoction and maceration method with 70% ethanol were used to extract the plant leaves. Phytochemical profiling of the extracts and compound identification was done using HPLC. Isoorientin was identified as the major phytochemical component of the extracts. MTT assay was used to screen cytotoxicity of the extract towards CRL2522, HaCaT, HepG2, MCF-7 and WRL68 cell lines. Combination index (CI) and isobologram were derived from combination treatment of extract and cyclophosphamide. Ethanolic extract has low cytotoxicity against all cell lines (IC50>1mg/mL). However, combination treatment of ethanolic extract and cyclophosphamide at IC15 and IC25 concentration resulted in synergism (CI

Perfluorinated compounds (PFCs) are non-biodegradable synthetic chemical compounds that are widely used in manufacturing many household products. Many studies have reported the association between PFCs exposure with the risk of developing cardiovascular diseases (CVDs). However, those reports are still debatable, due to their findings. Thus, this review paper aimed to analyse the association of PFCs compound with CVDs and their risk factors in humans by systematic review and meta-analysis. Google Scholar, PubMed and ScienceDirect were searched for PFCs studies on CVDs and their risk from 2009 until present. The association of PFCs exposure with the prevalence of CVDs and their risk factors were assessed by calculating the quality criteria, odds ratios (ORs), and 95% confidence intervals (CIs). CVDs risk factors were divided into serum lipid profile (main risk factor) and other known risk factors. The meta-analysis was then used to derive a combined OR test for heterogeneity in findings between studies. Twenty-nine articles were included. Our meta-analysis indicated that PFCs exposure could be associated with CVDs (Test for overall effect: z = 2.2, p = 0.02; Test for heterogeneity: I2 = 91.6%, CI = 0.92–1.58, p < 0.0001) and their risk factors (Test for overall effect: z = 4.03, p < 0.0001; Test for heterogeneity: I2 = 85.8%, CI = 1.00–1.14, p < 0.0001). In serum lipids, total cholesterol levels are frequently reported associated with the exposure of PFCs. Among PFCs, perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS) exposure increased the risk of CVDs than other types of PFCs. Although the risk of PFOA and PFOS were positively associated with CVDs and their risk factors, more observational studies shall be carried out to identify the long-term effects of these contaminants in premature CVDs development in patients.

BACKGROUND: The prevalence of non-alcoholic fatty liver disease (NAFLD) in Asian countries is increasing at concerning level. Currently, no specific treatment available to prevent its oxidative stress and progression except for diet and lifestyle changes. Vitamin E such as tocotrienol-rich fraction (TRF) has a promising potential in preventing NAFLD progression. TRF is a potent antioxidant but has low bioavailability due to the use of long-chain triglycerides (LCT) as its carrier. Testing of potential therapeutic agents such as TRF are commonly carried out using animal models. These animal models are often costly due to limited access to the supply especially Asian countries and predisposed to high transportation cost. Lower expenditure of NAFLD model should be investigated without forfeiting the outcome of study. Therefore, this study addresses the gap by utilizing the ICR mice as NAFLD model through dietary modification and testing on the newly formulated TRF with combination of palm kernel oil (PKO) as a medium-chain triglycerides (MCT) carrier. METHODS: Fifteen ICR strain mice were randomly group into two control and one treatment group. Control groups received high-fat diet (HFD) only and standard diet while treatment group was given HFD with TRF (200 mg/kg/day). Study was carried out for 10 weeks. Weights were recorded twice a week. At the end of study, all mice were euthanized and data such weights, waist circumference and random blood glucose were recorded. Liver from each mouse were prepared for histology assessment. RESULTS: Mice mean weights and random blood sugar showed no difference between group (P>0.05) while significance waist circumference was larger in HFD and TRF groups compared to SD (P

Bisphenol A, an endocrine disrupting compounds that affect human homeostasis. Studies on BPA are focusing on the impact of BPA in reproductive function and brain development. However, the effect of BPA on gut especially gastric cells is not well explored. Gut is directly in contact with ingested BPA; therefore, we aimed to determine the effect of BPA exposure on gastric cells proliferation at safe recommended concentration. Human gastric cancer cells (HGC-27) were treated with BPA at different concentration (low: 10 -9 M, 10 -7 M; high10 -5 M, 10 -4 M) and time point (24 hr, 48 hr, 72 hr). Cell viability assays were determined using MTS assay. Cells were further stained with Alexa Fluor-635 (F-actin) and Fluorescein (Hif-1α) protein for immunocytofluorescence. Data were analysed using ANOVA (p

The RNA binding protein Larp1 was originally shown to be involved in spermatogenesis, embryogenesis and cell-cycle progression in Drosophila. Our data show that mammalian Larp1 is found in a complex with poly A binding protein and eukaryote initiation factor 4E and is associated with 60S and 80S ribosomal subunits. A reduction in Larp1 expression by siRNA inhibits global protein synthesis rates and results in mitotic arrest and delayed cell migration. Consistent with these data we show that Larp1 protein is present at the leading edge of migrating cells and interacts directly with cytoskeletal components. Taken together, these data suggest a role for Larp1 in facilitating the synthesis of proteins required for cellular remodelling and migration.

RNA-binding proteins (RBPs) are increasingly identified as post-transcriptional drivers of cancer progression. The RBP LARP1 is an mRNA stability regulator, and elevated expression of the protein in hepatocellular and lung cancers is correlated with adverse prognosis. LARP1 associates with an mRNA interactome that is enriched for oncogenic transcripts. Here we explore the role of LARP1 in epithelial ovarian cancer, a disease characterized by the rapid acquisition of resistance to chemotherapy through the induction of pro-survival signalling. We show, using ovarian cell lines and xenografts, that LARP1 is required for cancer cell survival and chemotherapy resistance. LARP1 promotes tumour formation in vivo and maintains cancer stem cell-like populations. Using transcriptomic analysis following LARP1 knockdown, cross-referenced against the LARP1 interactome, we identify BCL2 and BIK as LARP1 mRNA targets. We demonstrate that, through an interaction with the 3′ untranslated regions (3′ UTRs) of BCL2 and BIK, LARP1 stabilizes BCL2 but destabilizes BIK with the net effect of resisting apoptosis. Together, our data indicate that by differentially regulating the stability of a selection of mRNAs, LARP1 promotes ovarian cancer progression and chemotherapy resistance.