HEV-Ag ELISA assay is a reliable diagnostic test in resource-limited areas. HEV genotype 1 (HEV-1) infections are either self-limited or progress to fulminant hepatic failure (FHF) and death if anti- HEV therapy is delayed. Limited data is available about the diagnostic utility of HEV Ag on HEV-1 infections. Herein we aimed to study the kinetics of HEV Ag during HEV-1 infections at different stages, i.e., acute HEV infection, recovery, and progression to FHF. Also, we evaluated the diagnostic utility of this marker to predict the outcomes of HEV-1 infections. Plasma of acute hepatitis E (AHE) patients were assessed for HEV RNA by RT-qPCR, HEV Ag, and anti-HEV IgM by ELISA. The kinetics of HEV Ag was monitored at different time points; acute phase of infection, recovery, FHF stage, and post-recovery. Our results showed that the level of HEV Ag was elevated in AHE patients with a significantly higher level in FHF patients than recovered patients. We identified a plasma HEV Ag threshold that can differentiate between self-limiting infection and FHF progression with 100% sensitivity and 88.89% specificity. HEV Ag and HEV RNA have similar kinetics during the acute phase and self-limiting infection. In the FHF stage, HEV Ag and anti-HEV IgM have similar patterns of kinetics which could be the cause of liver damage. In conclusion, the HEV Ag assay can be used as a biomarker for predicting the consequences of HEV-1 infections which could be diagnostically useful for taking the appropriate measures to reduce the complica tions, especially for high-risk groups.

A novel series of 2-substituted thio-7-chloroquinazolin-4(3H)-one derivatives was rationally designed, synthesized, and evaluated as dual inhibitors of epidermal growth factor receptor (EGFR) and vascular endothelial growth factor receptor-2 (VEGFR-2) with potential anticancer activity. Structural optimization was achieved through the incorporation of diverse linkers and pharmacophoric motifs, including aryl amides, hydrazones, chalcones, and heterocyclic moieties, to target key kinase binding domains. In vitro screening against the NCI-60 human tumor cell line panel revealed several compounds with broad-spectrum antiproliferative activity, among which the chalcone derivative (11a) emerged as the most potent. Compound (11a) exhibited low micromolar GI₅₀ values, high selectivity towards cancer cells over normal cells, and strong dual inhibition of EGFR and VEGFR-2 in the nanomolar range. Mechanistic studies demonstrated that (11a) induced G₁/S phase cell cycle arrest, activated apoptotic pathways, suppressed receptor phosphorylation, and promoted PARP-1 cleavage. Notably, (11a) retained inhibitory activity against clinically relevant EGFR resistance mutants, including C797S. Molecular docking and dynamics simulations indicated stable, favorable binding within the ATP-binding sites of both kinases. Collectively, these findings identify compound (11a) as a promising dual-target anticancer lead warranting further preclinical investigation.