Norazalina Saad

This work explores the potential of magnetite polymeric nanoparticles (MPNs) encapsulating Donepezil (DPZ) to evaluate its delivery potential in SH-SY5Y human neuroblastoma cells. The MNPs coated with oleic acid (OA) were synthesized via co-precipitation, resulting in oleic acid-coated magnetite NPs (OAMNPs). These were combined with Poly(lactic-co-glycolic acid) (PLGA) and DPZ to create the nanohybrid, OAMNPs-PLGA-DPZ. The nanohybrid has been characterized utilizing Dynamic light scattering (DLS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and Transmission electron microscopy (TEM). Drug loading capacity (LC%) and encapsulation efficiency (EE%) were quantified by Ultraviolet-Visible Spectrophotometry (UV-Vis), and in‑vitro drug release was evaluated at physiological pH 7.4. Finally, the cytotoxicity of OAMNPs-PLGA-DPZ on SH-SY5Y cells was determined using an MTT assay. The crystalline size of OAMNPs-PLGA-DPZ, as revealed by XRD, ranged from 20 to 60 nm, and the wurtzite crystal structure size was 12.95 nm. The FTIR spectra showed that DPZ was successfully encapsulated within the PLGA matrix, and the TEM images revealed the synthesis and production of OAMNP-PLGA-DPZ. Biocompatibility evaluation demonstrated an IC₅₀ of 13.5 ± 2.12 µg/mL as a baseline for the safety profile after 48 h of exposure. Taken together, these findings suggested that OAMNPs‑PLGA‑DPZ represent a foundational drug‑delivery platform with potential applications for neurodegenerative diseases.

Background: In traditional medicine, natural compounds, such as coumarins, have been recognised as effective therapeutic agents for a variety of ailments. One of the natural furanocoumarins is rutamarin, which can be abundantly found in the Rutaceae family. It has been demonstrated to possess diverse biological properties in preclinical models, underscoring its potential for therapeutic applications in various health problems. Main body: This review provides an extensive description and critical evaluation of the biological activities of rutamarin, particularly anticancer, antidepressant, antidiabetic, and antimicrobial effects. To a lesser extent, the antinociceptive, antivenom, and antiviral effects of rutamarin are also included. Scientific preclinical evidence based on in silico, in vitro, and in vivo models is discussed, highlighting its potential as a pharmaceutical agent for therapeutic purposes. As a natural product, the distribution and chemical extraction of rutamarin in plants, as well as its isolation, are also discussed. Moreover, efforts to amplify the yield of rutamarin in in vitro plant cultures and to synthesise rutamarin are briefly described. Since most studies previously utilised in silico analysis to elucidate rutamarin’s mechanism of action, limitations and future directions for clinical trials are also suggested. Conclusion: Advancement to clinical trials is a crucial step towards utilising rutamarin as a therapeutic agent for various diseases and health complications, including cancers, diabetes, and microbial infections. However, current findings that largely rely on in silico and in vitro models need further preclinical validation of the biological properties, especially in vivo models, which are essential prior to advancement to clinical setting.

RNA interference is crucial in post-transcriptional gene silencing. Short hairpin RNA (shRNA) is particularly effective because it forms fully complementary matches with target mRNA, leading to its degradation. However, shRNA processing relies on nuclear microprocessors like Drosha, posing a challenge for RNA viral vectors that replicate exclusively in the cytoplasm. Although there have been reports of Drosha translocating to the cytoplasm upon viral infection, many RNA viruses, including Newcastle disease virus (NDV), remain inadequately studied in this context and, in some cases, fail to induce Drosha translocation for shRNA processing. In this study, we developed a novel approach to translocate an shRNA, expressed by NDV as an RNA viral vector, into the nucleus for Drosha processing. As a proof of concept, a recombinant NDV expressing the shRNA (rAF-shmcherry) with an AU-rich region at its 3′ end in the expression cassette was constructed. This shRNA targets a constitutively expressed mCherry gene in a colorectal cancer cell line, SW620-mC. We confirmed the presence of the AU-rich shRNA in the nuclei of the rAF-shmcherry-infected SW620-mC using reverse transcription PCR (RT-PCR). The gene-silencing effect of the shRNA was then evaluated at mRNA and protein levels, showing ~90% downregulation of the mCherry transgene at 24 h post-infection and 70% downregulation of mCherry protein in SW620-mC at 48 h post-infection. This study marks the first exploration of NDV as an shRNA viral vector, presenting a promising approach for shRNA translocation that could be applicable to various RNA viruses.

Introduction: Breast cancer remains a significant challenge due to disease recurrence and metastasis, often attributed to the persistence of treatment-resistant breast cancer stem cells (BCSCs). Citral, a compound derived from lemongrass essential oil, has demonstrated cytotoxic effects on various cancers, including breast cancer. This study investigates the anticancer mechanisms of Citral in BCSC-enriched 3D cultured spheroids and evaluates its therapeutic potential for estrogen receptor-positive breast cancer. Methods: Using flow cytometry, the CD44+CD24- population was analyzed, and real-time PCR was employed to measure the expression of ALDH isoforms and pluripotency genes. Additionally, the Nanostring nCounter® PanCancer Pathway Panel was utilized to identify gene expression changes in cancer-related pathways. Results: Citral treatment significantly reduced spheroid size and the CD44+CD24- stem-like cell population, accompanied by downregulation of ALDH isoforms and pluripotency genes. Gene expression analysis revealed Citral's modulation of key pathways, including PI3K/Akt signaling, cell cycle control, DNA damage response, and apoptosis. Conclusion: These findings underscore Citral's potential as a promising anticancer agent, particularly for targeting estrogen receptor-positive breast cancer cells and BCSCs. Further preclinical and clinical studies are warranted to explore its therapeutic applications.

Postbiotic metabolites produced by Lactiplantibacillus plantarum strains isolated from Malaysian food have been extensively reported for their positive effects on health. Understanding the effects of different combinations of carbon and nitrogen sources on the growth and corresponding characteristics of postbiotic metabolites produced by different strains of L. plantarum is important for various applications. Hence, the effects of different combinations of carbon (glucose, lactose, sucrose and dextrose) and nitrogen (X-SEED Kat, X-SEED Peptone, X-SEED Nucleo Advanced, Nucel875 MG, FM888 and FM902) sources on the growth of six strains of L. plantarum (RG11, RG14, RI11, RS5, TL1 and UL4) and the functional characteristics (bacteriocin inhibitory activity, antioxidant activity and lactic acid concentration) of their respective postbiotic metabolites were investigated in this study. UL4 produced the highest viable cell population with sucrose and Nucel875 nitrogen source. The UL4 strain also produced the strongest bacteriocin inhibitory activity with dextrose and FM888 nitrogen source. In comparison, the RI11 strain produced the highest lactic acid concentration with dextrose and Nucel875 nitrogen source and the highest reducing power of RS5 and TL1 postbiotic metabolites was achieved with MRS medium. In the combination of sucrose and X-Seed KAT nitrogen source, RG14 produce the highest hydroxyl radical scavenging activity. The effects of different combinations of carbon and nitrogen sources on the viable cell population of L. plantarum strains and the respective functional characteristics of postbiotic metabolites were strain dependent. The current study also revealed that fermentation media were an important factor that greatly impacted the functionalities of postbiotic metabolites due to the presence of various bioactive compounds that contributed to high antioxidant and antimicrobial properties. The results of this study will facilitate the subsequent medium design and optimisation for the development and production of specific postbiotic metabolites produced by the respective L. plantarum strain for their applications in various industries.

Matrix metallopeptidase 3 (MMP3) is among the key players in breast cancer metastasis that contributes to the highest cancer-related deaths in females globally. Previously, in silico analyses had shown that several coding single nucleotide polymorphisms (SNPs) of MMP3 were predicted to alter the secondary structures of MMP3 and subsequently reduce its mRNA stability. To validate the mentioned hypotheses, this study aimed to determine the effects of six coding SNPs of MMP3 on its mRNA stability, protein expression level as well as cell invasiveness in vitro. In this study, breast adenocarcinoma MCF7 cells were transfected with MMP3 wild type (MMP3-WT) and a variant containing SNPs (MMP3-Var). Following transfection, protein expression level, mRNA stability and enzyme activity of MMP3-WT and MMP3-Var were evaluated. Finally, the effect of MMP3 coding SNPs on cell invasiveness in breast cancer was determined. In this study, the mRNA stability, protein expression level and enzymatic activity of MMP3-Var were significantly reduced. Moreover, the presence of MMP3 coding SNPs led to attenuated invasiveness of transfected MCF7 cells. In conclusion, these findings may contribute to the current understanding of these coding SNPs with metastasis in breast cancer.

Tuberculosis (TB) stands as an enduring and formidable global health challenge, necessitating innovative methodologies to optimize therapeutic outcomes. Within this context, nanotechnology emerges as a promising modality to augment TB therapy by enabling targeted drug delivery of anti-TB agents to infect cells, thereby maximizing therapeutic efficacy while mitigating adverse effects. This study focalizes on the synthesis of chitosan nanoparticles (CSNP) as a nanocarrier for the concurrent delivery of Pyrazinamide (PYR), a pivotal antibiotic in the first-line treatment of TB. The primary objective is to overcome challenges related to the limited solubility and bioavailability of these drugs, which hinder their effectiveness and pose toxicity risks. The encapsulation of PYR within CS proves advantageous, enhancing solubility, bioavailability, and targeted drug delivery. The resulting nanocarrier, denoted as CS-PYR, is prepared through ionic gelation, involving cross-linking chitosan-loaded anti-TB carriers with tripolyphosphate to achieve nanoscale dimensions. Various analytical techniques, including XRD, FTIR, TGA, TEM, FESEM, DLS, UV-vis spectroscopy, antibacterial assays, and cytotoxic MTT assays, are utilized for a comprehensive investigation. The spherical morphology of PYR-CSNP, with an average diameter of 60 to 100 nm, is confirmed by FESEM, TEM, and DLS analyses. FTIR and XRD analyses validate the successful fabrication and multifunctional properties of PYR incorporated into the CSNP matrix. The drug-loaded CSNP exhibits improved thermal stability, displaying decomposition between 200-400°C, indicating higher temperature resistance and greater weight loss compared to the CSNP carrier due to PYR inclusion. Drug release studies show sustained release behaviour, with 90-99% release in an acidic (pH 4.8) buffer solution over seven days, following a pseudo-second-order model. Moreover, the CS-PYR nanocarrier demonstrates superior efficacy against Gram-negative and Gram-positive bacteria, attributed to drug intercalation resulting in larger inhibition zones. Cytotoxicity assessments for CS-PYR indicate 80% cell viability in MRC5 cells, underscoring the nanocarrier's potential as a safe and effective TB treatment delivery system without adversely affecting normal cells. These findings position CS-PYR as a promising nanodelivery system for TB therapy, offering enhanced precision in drug delivery and safety.

Neuronal models are an important tool in neuroscientific research. Hydrogen peroxide (H₂O₂), a major risk factor of neuronal oxidative stress, initiates a cascade of neuronal cell death. Polygonum minus Huds, known as ‘kesum’, is widely used in traditional medicine. P. minus has been reported to exhibit a few medicinal and pharmacological properties. The current study aimed to investigate the neuroprotective effects of P. minus ethanolic extract (PMEE) on H₂O₂-induced neurotoxicity in SH-SY5Y cells. LC–MS/MS revealed the presence of 28 metabolites in PMEE. Our study showed that the PMEE provided neuroprotection against H₂O₂-induced oxidative stress by activating the Nrf2/ARE, NF-κB/IκB and MAPK signaling pathways in PMEE pre-treated differentiated SH-SY5Y cells. Meanwhile, the acetylcholine (ACH) level was increased in the oxidative stress-induced treatment group after 4 h of exposure with H₂O₂. Molecular docking results with acetylcholinesterase (AChE) depicted that quercitrin showed the highest docking score at −9.5 kcal/mol followed by aloe-emodin, afzelin, and citreorosein at −9.4, −9.3 and −9.0 kcal/mol, respectively, compared to the other PMEE’s identified compounds, which show lower docking scores. The results indicate that PMEE has neuroprotective effects on SH-SY5Y neuroblastoma cells in vitro. In conclusion, PMEE may aid in reducing oxidative stress as a preventative therapy for neurodegenerative diseases.

Since lung cancer is the leading cause of cancer-related death worldwide, research is being conducted to discover anticancer agents as its treatment. Eleutherine bulbosa, a Dayak folklore medicine, exhibited anticancer effects against several cancer cells; however, its anticancer potency against lung cancer cells has not been explored yet. This study aims to determine the anticancer potency of E. bulbosa bulbs against lung cancer cells (A549) using 2D and 3D culture models, as well as determine its active compounds using gas chromatography-mass spectrometry (GC-MS) analysis. Three fractions of E. bulbosa bulbs, namely chloroform, n-hexane, and ethyl acetate, were tested for cytotoxicity using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) and CellTiter-Glo. The antiproliferative effects of the most cytotoxic fraction against the 2D culture model were determined by a clonogenic survival assay and propidium iodide/Hoechst 33342 double staining, whereas the effects against the 3D culture model were determined by microscopy, flow cytometry, and gene expression analysis. The chloroform fraction is the most cytotoxic against A549 cells than other fractions, and it inhibited colony formation and induced apoptosis of A549 cells. The chloroform fraction also inhibited the growth of the A549 spheroid by suppressing the spheroid size, inducing apoptosis, reducing the proportion of CD44 lung cancer stem cells, causing arrest at the S phase of the cell cycle, and suppressing the expression of the SOX2 and MYC genes. Furthermore, the GC-MS analysis detected 20 active compounds in the chloroform fraction, including the major compounds of eleutherine and isoeleutherine. In conclusion, the chloroform fraction of E. bulbosa bulbs exhibit its antiproliferative effect on 2D and 3D culture models of A549 cells, suggesting it could be a lung cancer chemopreventive agent.