Lacticaseibacillus paracasei is one of the probiotic bacteria widely identified from fermented foods. The application of L. paracasei is commonly used in dairy and non-dairy products. To investigate the probiotic properties of L. paracasei cells including their acid, pepsin, pancreatin, and bile salt tolerances; adhesion ability; antipathogen activity; and antibiotic susceptibility, L. paracasei cells were incorporated into skim milk and lyophilized by freeze drying. Freeze-dried probiotic cells were add to green banana powder and low moisture additive food matrices and a storage analysis of the product was performed. The result showed that L. paracasei cells possessed potentially beneficial probiotic properties to survive stress in the gastrointestinal tract (GIT) and functional abilities as an anti-enteropathogenic agent; they were also safe to use and displayed antibiotic properties. Furthermore, the probiotic freeze-drying technique preserved high probiotic cell survivability (10¹¹ CFU/g). In term of prolonged storage (60 days), the powder product was stable and maintained probiotic survival (10⁷ CFU/g) while excluding non-probiotic growth. In conclusion, L. paracasei displayed probiotic properties in the GIT and was judged to be a highly acceptable product as a probiotics–banana rehydrated beverage.

Triple negative breast cancer (TNBC) is a breast cancer subtype characterized by the absence of estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2 expression. TNBC cells respond poorly to targeted chemotherapies currently in use and the mortality rate of TNBC remains high. Therefore, it is necessary to identify new chemotherapeutic agents for TNBC. In this study, the anti-cancer effects of 7-α-hydroxyfrullanolide (7HF), derived from Grangea maderaspatana, on MCF-7, MDA-MB-231 and MDA-MB-468 breast cancer cells were assessed using MTT assay. The mode of action of 7HF in TNBC cells treated with 6, 12 and 24 µM of 7HF was determined by flow cytometry and propidium iodide (PI) staining for cell cycle analysis and annexin V/fluorescein isothiocyanate + PI staining for detecting apoptosis. The molecular mechanism of action of 7HF in TNBC cells was investigated by evaluating protein expression using proteomic techniques and western blotting. Subsequently, 7HF exhibited the strongest anti-TNBC activity toward MDA-MB-468 cells and a concomitantly weak toxicity toward normal breast cells. The molecular mechanism of action of low-dose 7HF in TNBC cells primarily involved G2/M-phase arrest through upregulation of the expression of Bub3, cyclin B1, phosphorylated Cdk1 (Tyr 15) and p53-independent p21. Contrastingly, the upregulation of PP2A-A subunit expression may have modulated the suppression of various cell survival proteins such as p-Akt (Ser 473), FoxO3a and β-catenin. The concurrent apoptotic effect of 7HF on the treated cells was mediated via both intrinsic and extrinsic modes through the upregulation of Bax and active cleaved caspase-7–9 expression and downregulation of Bcl-2 and full-length caspase-7–9 expression. Notably, the proteomic approach revealed the upregulation of the expression of pivotal protein clusters associated with G1/S-phase arrest, G2/M-phase transition and apoptosis. Thus, 7HF exhibits promising anti-TNBC activity and at a low dose, it modulates signal transduction associated with G2/M-phase arrest and apoptosis.

Background and Aim: Malaria continues to pose a global health challenge, exacerbated by the emergence of drug-resistant strains of Plasmodium falciparum. This study aimed to evaluate the anti-Plasmodium potential of Propolis extracts collected from various Iranian regions and to characterize the molecular interactions of their bioactive phytochemicals with P. falciparum lactate dehydrogenase (PfLDH), a key enzyme in parasite glycolysis. Materials and Methods: The anti-Plasmodium activity of ethanol-extracted Propolis was assessed against P. falciparum NF54 using the SYBR Green I fluorescence assay. Gas chromatography-mass spectrometry (GC-MS) analysis identified major phytochemicals in the most active extract. Molecular docking and 100-ns molecular dynamic (MD) simulations were performed to evaluate the binding affinity and stability of selected compounds (tectochrysin and galangin) against PfLDH in both holo (Protein Data Bank [PDB] ID: 1LDG) and apo (PDB ID: 2X8L) forms. Results: Propolis collected from Kermanshah city exhibited the highest anti-Plasmodium activity (IC50 = 6.69 ± 1.44 μg/mL). GC-MS analysis identified tectochrysin and galangin as major constituents. Molecular docking revealed strong binding affinities of tectochrysin (−7.8 kcal/mol) and galangin (−7.5 kcal/mol) to PfLDH, surpassing the binding energies of standard antimalarial drugs (chloroquine and quinine). MD simulations confirmed the stability of tectochrysin and galangin within the PfLDH active sites, with favorable root mean square deviation, root mean square fluctuation, gyration, solvent-accessible surface area, molecular surface area, and polar surface area profiles, indicating persistent and stable protein-ligand interactions throughout the simulation. Conclusion: The findings support the promising anti-Plasmodium potential of Propolis-derived compounds, particularly tectochrysin and galangin, as novel PfLDH inhibitors. Their potential applicability in transdisciplinary anti-parasitic therapy across human and veterinary medicine warrants further in vivo validation and clinical investigations. Keywords: anti-malarial candidates, galangin, lactate dehydrogenase, molecular docking, molecular dynamics, Plasmodium falciparum, Propolis extract, tectochrysin

Background and Aim: Acanthamoeba, an opportunistic protozoan, exists widely in natural sources and can cause infections in humans and animals. The absence of effective monotherapy after the initial infection leads to chronic disease and recurrence. Tubulin protein is a vital target for design-targeted drug discovery. Anti-tubulin drugs are also used to treat Acanthamoeba infection, although resistance to these drugs has been observed. Therefore, it is necessary to identify a new targeted drug for Acanthamoeba infection. Therefore, this study aimed to assess the in vitro activity of ethanol extracts of Rhizophora apiculata extracts (RAE) against Acanthamoeba spp. and to predict its chemical compound on β-tubulin interaction. Materials and Methods: Acanthamoeba, an opportunistic protozoan, exists widely in natural sources and can cause infections in humans and animals. The absence of effective monotherapy after the initial infection leads to chronic disease and recurrence. Tubulin protein is a vital target for design-targeted drug discovery. Anti-tubulin drugs are also used to treat Acanthamoeba infection, although resistance to these drugs has been observed. Therefore, it is necessary to identify a new targeted drug for Acanthamoeba infection. Therefore, this study aimed to assess the in vitro activity of ethanol extracts of Rhizophora apiculata extracts (RAE) against Acanthamoeba spp. and to predict its chemical compound on β-tubulin interaction. Results: The results revealed that A. triangularis and A. polyphaga trophozoites had the highest inhibition at 90% at a MIC of 8 mg/mL after treatment with RALE and RABE, respectively, at 24 h. Those MPC values were exhibited at 16 mg/mL against A. triangularis trophozoites. In addition, both extracts inhibited the adhesive properties of all Acanthamoeba approximately 80%–90% at 4 mg/ mL, as well as adherent structural acanthopodia loss. MCC was 0.25 mg/mL, provided to be harmless to mammalian cells. GC-MS analysis supported that 8 and 11 major phytochemicals were from RABE and RALE, respectively. Molecular docking and molecular dynamics demonstrated that Acanthamoeba-β-tubulin exhibited potent root-mean-square deviation, root mean square fluctuation, and binding free energy values with clionasterol (from RABE and RALE) and stigmasterol (from RALE). Based on our results, ethanolic RABE and RALE exhibited anti-Acanthamoeba activity in reducing adhesion. In silico showed that promising clionasterol and stigmasterol interacted with a targeting β-tubulin. Conclusion: The RABE and RALE exhibited a potential adherent effect on A. triangularis, low toxicity, and the clionasterol and stigmasterol in RABE and RALE predicted to interact the targeted β-tubulin. These agents may be used as alternative therapeutic agents in the management of disease using a sustainable one-heath approach. Keywords: Acanthamoeba triangularis, anti-adherent activity, molecular docking, molecular dynamic simulation, Rhizophora, tubulin.

Background and Aim: Acanthamoeba spp. is free-living protozoa capable of causing severe infections, notably Acanthamoeba keratitis, which is difficult to manage due to cyst resistance and the cytotoxicity of current treatments. Plant-derived com­pounds represent a promising alternative strategy. This study investigated the amoebicidal, anti-adhesive, and cytotoxic properties of Mangifera indica L. (mango) leaf extract against ocularly relevant Acanthamoeba spp. Materials and Methods: Crude ethanolic leaf extract of M. indica was prepared and evaluated against Acanthamoeba polyphaga American Type Culture Collection (ATCC) 30461 and Acanthamoeba castellanii ATCC 50739. Minimum inhibitory concentration (MIC) and minimum parasiticidal concentration were determined for trophozoites and cysts. Morphological changes were analyzed by scanning electron microscopy (SEM). Anti-adhesion assays were conducted using polystyrene surfaces, with a commercial multipurpose contact lens (CL) solution as a control. Cytotoxicity was tested in Vero cells using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide assay to establish the minimum cytotoxic concentration. Results: The extract inhibited trophozoite growth at 2 mg/mL and demonstrated cysticidal activity at 4 mg/mL for A. polyph­aga and 32 mg/mL for A. castellanii. SEM revealed disruption of trophozoite morphology, loss of acanthopodia, and sur­face perforations in cysts. At MIC levels, adhesion was reduced by >70%, and even at 1/8 MIC, inhibition remained above 50%, comparable to a commercial multipurpose solution. Cytotoxicity assessment showed >80% Vero cell viability at 0.125 mg/mL, indicating a favorable therapeutic window. Conclusion: This is the first report demonstrating amoebicidal and anti-adhesive effects of M. indica L. leaf extract against ocular Acanthamoeba species. The dual trophozoiticidal and anti-adhesive actions, combined with low cytotoxicity, high­light its potential for development as a plant-based therapeutic agent, particularly in ocular formulations or CL disinfectants. Future work should focus on phytochemical isolation, mechanistic studies, and novel delivery systems to enhance efficacy and safety. Keywords: Acanthamoeba, amoebicidal activity, anti-adhesion, cytotoxicity, Mangifera indica L., ocular infection.

Acanthamoeba spp., free‐living amoebae (FLA), are the causative agents of Acanthamoeba keratitis (AK) and granulomatous amoebic encephalitis (GAE). These diseases have limited treatment options; therefore, drug discovery remains a critical priority. This study evaluated the leaf extract of Annona muricata L., which contains quercetin, formulated in silica nanoparticles against Acanthamoeba species. The anti‐ Acanthamoeba effect of the nanoformulation was determined by its MIC and percentage viability. Scanning electron microscopy confirmed the nanocompound‐directed effect on trophozoites and cysts. The nanoquercetin exhibited anti‐ Acanthamoeba activity, showing greater effectiveness against the trophozoites of Acanthamoeba polyphaga ATCC 30461 (AP) (MIC > 2 mg/mL) compared to cysts (MIC > 8 mg/mL) and against A. triangularis WU19001 (AT) trophozoites (MIC > 4 mg/mL) compared to cysts (MIC > 8 mg/mL). It also showed reduced activity against Acanthamoeba castellanii ATCC 30010 (AC3) trophozoites (MIC > 4 mg/mL) compared to cysts (MIC > 8 mg/mL) and A. castellanii ATCC50739 (AC5) trophozoites (MIC > 4 mg/mL) compared to cysts (MIC > 8 mg/mL). The anti‐ Acanthamoeba activity of A. muricata on all four species was compared to chlorhexidine, which had MIC values of 0.008 mg/mL for AC3 and AT trophozoites and 0.016 mg/mL for AC5 and AP trophozoites. For cysts, chlorhexidine exhibited MIC values of > 0.0128 mg/mL for AC3 and AP, 0.032 mg/mL for AC5, and 0.064 mg/mL for AT. All MIC values demonstrated inhibition of Acanthamoeba species with reduced viability exceeding 90%. SEM findings revealed morphological changes in Acanthamoeba , indicating parasitic inhibition. Quercetin‐loaded silica nanoparticles demonstrated anti‐ Acanthamoeba activity with low cytotoxicity. This indicates the potential of quercetin as a nano‐enhanced natural therapeutic or contact lens‐care agent. This study is limited by its in vitro design, small parasite strain panel, and potential nanoparticle variability. This highlights the need for in vivo validation, ocular/contact lens model testing, and formulation optimization for improved cysticidal efficacy.