The extensive evolutionary diversity and continuous mutational adaptation of betacoronaviruses represent a major challenge to the development of broadly protective countermeasures. This study applied an evolution-informed Viroinformatics framework to systematically identify conserved and immune targets with predicted lower mutational vulnerability across betacoronaviruses. The workflow integrated comparative sequence conservation and phylogenetic analysis with immunoinformatics-based epitope screening, HLA population coverage, mutational escape profiling, structural modeling, molecular docking, and molecular dynamics simulations. Conserved cytotoxic T-lymphocyte and helper T-lymphocyte epitopes were prioritized based on immunogenicity, antigenicity, safety, HLA binding potential, evolutionary conservation, and predicted resistance to mutational escape. Combined MHC class I and II analysis predicted a theoretical global population coverage of 99.92%, which requires experimental validation. The resulting 409-amino-acid multiepitope construct was predicted to be antigenic, non-allergenic, and physiochemically stable. Structural validation showed that 99.7% of the residues were located within favored or additionally allowed regions. Molecular docking demonstrated favorable interaction of the vaccine construct with TLR7, while a 100 ns molecular dynamics simulation supported persistence of the predicted vaccine-receptor association. Overall, these findings provide a computational framework for prioritizing evolutionarily conserved immune targets and developing pre-emptive candidate epitopes showing predicted cross-betacoronavirus conservation within the analyzed dataset. Experimental studies evaluating HLA binding, cellular and humoral immunogenicity, recombinant expression, cross-reactivity, and in vivo protection are required to validate the predicted immunological potential and determine whether the proposed construct can provide broad protection against diverse betacoronaviruses.
Ethanol has variable effectiveness against non-enveloped viruses; however, several reports suggest that it possesses virucidal activity against human adenoviruses (HAdVs). Ethanol concentrations between 45 w/w% and 95 w/w% showed sufficient activity against HAdV type 5 over a 30 s period, while concentration between 72.5 w/w% and 77 w/w% was effective against type 7 over 60 s period. Type 2 and 8 were less susceptible, however. In this study, we investigated the ability of different concentrations of ethanol to inactivate and change the structure of 19 HAdV types across six subgroups, using the virus infectivity titer for evaluation. After 1 min of ethanol treatment, HAdV types 5 and 8 were observed with transmission electron microscopy. The virucidal activity of disinfectant ethanol against HAdVs in subgroups A, B1 (except for type 21), and C was higher than that in subgroups B2, D, E, and F. We observed a ‘U-shaped’ fluctuation pattern where the reduction factor dropped at concentrations of 70–80% and then rose again at 90%. After treatment with 40% ethanol, TEM images showed no major changes in type 8 HAdVs, but organic matter was observed around the virus particles in type 5 HAdVs; after treatment with 66% ethanol, virus particles aggregated in both types. This suggests that HAdVs with a lower number of repeats in the shaft region exhibited higher ethanol resistance.
As domestic and international travel has expanded, the risk of the cross-border dissemination of infectious pathogens has increased, as clearly evidenced during the COVID-19 pandemic. Aircraft- and airport-based wastewater surveillance can provide timely, aggregate evidence of pathogen circulation among mobile populations and may complement clinical, genomic, and syndromic surveillance at points of entry. The strongest evidence for this approach comes from SARS-CoV-2 RNA detection and variant monitoring in aircraft lavatory wastewater, airport terminal wastewater, and airport-associated wastewater systems. However, aviation wastewater signals require cautious interpretation because detectability depends on pathogen shedding dynamics, passenger lavatory use, flight duration, sampling matrix, sample volume, assay sensitivity, sequencing depth, and the availability of epidemiological metadata. This structured narrative review synthesizes current evidence on aircraft and airport wastewater surveillance and examines the scientific determinants that influence surveillance validity and interpretation. Key methodological considerations include sampling design, sample preservation, nucleic acid extraction, molecular detection, sequencing, bioinformatics, inhibition assessment, recovery controls, assay sensitivity, and quality assurance. The review emphasizes that wastewater signals are aggregate and matrix-dependent and should not be interpreted as individual-level diagnosis, direct evidence of transmission, or definitive attribution to a specific flight, route, country, or population. Although SARS-CoV-2 surveillance is comparatively well-documented, multi-pathogen surveillance for enteric viruses, respiratory viruses, vaccine-preventable infections, emerging pathogens, and antimicrobial resistance markers requires pathogen-specific validation and clearer interpretation criteria. Aircraft and airport wastewater surveillance is therefore best viewed as an early-warning and situational-awareness approach whose value depends on converting technical signals into scientifically interpretable evidence.
Professor Robin Weiss, who died earlier this year, made profound early contributions to how HIV-1 and related immunodeficiency viruses infect cells via the CD4 receptor, and the role of neutralising antibodies in inhibiting HIV-1 infection. These and many other scientific contributions, alongside his mentorship of multiple scientists over decades, continue to impact today’s research and maintain his memory.
Hepatitis C Virus (HCV) is a positive strand RNA virus that infects hepatocytes and reprograms cellular gene expression, contributing to chronic liver disease and hepatocellular carcinoma. HCV replicates slowly and induces limited changes in the cellular transcriptome. Here, we improved analysis of low-abundance transcripts, including long non-coding RNAs (lncRNAs), in the context of HCV replication. We stringently removed genomic DNA from total RNA from Huh-7 hepatoma cell lines by a combination of TRIzol, acidic phenol, silica matrix columns and recombinant DNase I. RNA sequencing (RNA-Seq) and targeted validation were used to characterize Lnc-ITM2C-1 splice variants and expression of its neighboring gene, G protein-coupled receptor 55 (GPR55), to investigate their potential roles in HCV replication. Challenging our previous findings, under our improved experimental conditions, Lnc-ITM2C-1 and GPR55 were neither involved in a positive regulation circuit with HCV nor induced detectable transcriptome changes. Secondary analysis of RNA-Seq data according to HCV infection status revealed statistically robust HCV-induced expression of 21 host cell genes involved in pathways like stress response (in particular ATF3), lipid metabolism, cytoskeleton organization and growth regulation. In contrast, we found that expression changed for only a limited subset of previously reported HCV-associated lncRNAs (e.g., EGOT). This highlights the necessity of stringent gDNA removal for analysis of low-abundance transcripts like lncRNAs.
