Objectives: Ellagic acid (EA) has a wide range of biological effects. The purpose of this study was to investigate the in vitro effects of EA on HIV-1 replication, viral enzyme activity and cytokine secretion by infected cells. Methods: The anti-HIV-1 activity of EA in solution was determined in vitro using the infection of TZM-bl cells by the nano luciferase-secreting R5-tropic JRCSF strain of HIV-1, which allows for the quantification of viral growth by measuring nano luciferase in the culture supernatants. The effect of EA on the cytokine secretion of TZM-bl cells was determined by a multiplexed bead array after 48 h of HIV-1 exposure. The antiviral effect of EA in the gel formulation (Ellagel), as would be used for vaginal application, was investigated by the inhibition of infection of UC87.CD4.CCR5 cells with R5-tropic pBaLEnv-recombinant HIV-1. Results: EA in solutions of up to 100 µM was not toxic to TZM-bl cells. EA added either 1 h before or 4 h after HIV-1 exposure suppressed the replication of R5-tropic HIV-1 in TZM-bl cells in a dose-dependent manner, with up to 69% inhibition at 50 µM. EA-containing solutions also exhibited a dose-dependent inhibitory effect on HIV-1 replication in U87 cells. When EA was formulated as a gel, Ellagel containing 25 µM and 50 µM EA inhibited HIV-1 replication in U87 cells by 56% and 84%, respectively. In assays of specific HIV-1 enzyme activity, Ellagel inhibited HIV-1 integrase but not protease. EA did not significantly modulate cytokine secretion. Conclusions: We conclude that EA either in solution or in a gel form inhibits HIV infection without adverse effects on target cells. Thus, gel containing EA can be tested as a new microbicide against HIV infection.

Ellagic acid (EA) is a phenolic phytochemical found in many plants and their fruits. Vaginal epithelial cells are the first line of defense against pathogen invasion in the female reproductive tract and express antimicrobial peptides, including hBD2 and SLPI. This study investigated the in vitro effects of EA (1) on vaginal innate immunity using human vaginal epithelial cells, and (2) on HPV16 pseudovirus infection. Vaginal cells were cultured in the presence or absence of EA, and the expression of hBD2 and SLPI was determined at both transcriptional and translational levels. In addition, secretion of various cytokines and chemokines was measured. Cytotoxicity of EA was determined by CellTiter-blue and MTT assays. To investigate the ability of EA to inhibit HPV16 infection, EA was used to treat HEK-293FT cells in pre-attachment and adsorption steps. We found significant increases in both hBD2 mRNA (mean 2.9-fold at 12.5 µM EA, p < 0.001) and protein (mean 7.1-fold at 12.5 µM EA, p = 0.002) in response to EA. SLPI mRNA also increased significantly (mean 1.4-fold at 25 µM EA, p = 0.01), but SLPI protein did not. Secretion of IL-2 but not of other cytokines/chemokines was induced by EA in a dose-dependent manner. EA was not cytotoxic. At the pre-attachment step, EA at CC₂₀ and CC₅₀ showed a slight trend towards inhibiting HPV16 pseudovirus, but this was not significant. In summary, vaginal epithelial cells can respond to EA by producing innate immune factors, and at tested concentrations, EA is not cytotoxic. Thus, plant-derived EA could be useful as an immunomodulatory agent to improve vaginal health.

Purpose: This study was designed to investigate anticancer effect of ethanol and aqueous extracts of Lygodium microphyllum (Cav.) R.Br., Lygodium salicifolium Presl, and Licuala longecalyculata Furtado on human lung adenocarcinoma A549 cell line, and to determine total phenolic and flavonoid contents of those medicinal plants. Methods: The three medicinal plants were extracted with ethanol and water using a rotary evaporator and freeze dryer, respectively. Cytotoxicity of the crude extracts at concentrations ranging from 3.125 to 200 μg/ml on A549 cells was determined using MTT assay at various times. Aluminium chloride colorimetric assay and the Folin-Ciocalteu colorimetric method were used to analyze total flavonoid and phenolic content, respectively. Results: L. longecalyculata Furtado extracted with ethanol strongly inhibited A549 cell proliferation in a dose- and time-dependent manner with an IC₅₀ value of 12.00±0.41 μg/ml, whereas L. microphyllum (Cav.) R.Br. and L. salicifolium Presl did not. In addition to the morphological changes, an anticancer effect was observed in L. longecalyculata Furtado-treated A549 cells. L. longecalyculata Furtado showed the presence of phenolic and flavonoid compounds. Conclusion: These results indicate that in accordance with local wisdom, L. longecalyculata Furtado crude extract can empirically inhibit lung cancer cell line, but L. microphyllum (Cav.) R.Br. and L. salicifolium Presl cannot. However, further study of L. longecalyculata Furtado is needed to identify phytochemical compounds and their mechanism of anticancer action, in order to develop them as a new therapeutic agent for lung cancer management.

Currently, no validated population-based screening strategy uses persistent oral high-risk human papillomavirus (HPV) detection to identify individuals at clinically meaningful risk of HPV-associated oropharyngeal squamous cell carcinoma (OPSCC). This narrative review evaluates sampling strategies, virologic assays, liquid-biopsy biomarkers, adjunct biomarkers, and integrative approaches across three clinical contexts: longitudinal oral HPV assessment; diagnostic evaluation and HPV attribution in suspected or newly diagnosed OPSCC; and treatment monitoring and post-treatment surveillance in established HPV-associated OPSCC. The literature published from January 2020 through June 2026 and identified in PubMed was synthesized by clinical context, specimen source, biological target, study population, and intended use. Repeated detection of the same high-risk HPV genotype in serial oral specimens provides a pragmatic measure of persistent oral HPV detection but does not establish uninterrupted infection, sustained viral oncogene transcription, malignant transformation, or future OPSCC risk. Oral HPV DNA, oral E6/E7 mRNA, tumor-tissue HPV testing, and plasma circulating tumor HPV DNA represent distinct biological and clinical signals. Plasma circulating tumor HPV DNA has the most mature evidence for treatment monitoring and post-treatment surveillance in established HPV-associated OPSCC, not population screening. Clinical translation requires standardized longitudinal protocols, validation in intended-use populations, actionable management pathways, and incremental predictive value for proposed risk-stratification models.