The most prevalent RNA modification in eukaryotic cells is the addition of N6-methyladenosine (m6A) to RNA, which plays a critical role in regulating RNA stability and translation co-transcriptionally. These modifications are reversibly catalyzed by m6A methyltransferases (writers), demethylases (erasers), and utilized by m6A-binding proteins (readers). The hepatitis E virus (HEV) has a single-stranded, positive-sense (+) RNA genome that undergoes cap-dependent translation. However, the host factors involved in the replication cycle of HEV, particularly RNA modifications that occur during viral replication, remain poorly understood. This study aims to explore the roles of m6A writers, readers, and erasers in the context of HEV infection. To assess the impact of m6A RNA modifications on HEV infection, we administered the methyltransferase METTL3 inhibitor STM2457 to HEV-infected cells in a dose-dependent manner, with subsequent screening for virus-positive cells using fluorescence microscopy. Additionally, HEV replicon-transfected cells were treated with the inhibitor to evaluate effects on viral replication via luciferase assay. Our findings indicate that inhibiting m6A methylation through the use of STM2457 disrupts infection with the human-derived HEV-3 strain Kernow-C1/p6 at an EC50 of ~ 10 nM. However, viral replication remains unaffected when in vitro transcribed RNA is transfected by electroporation. Moreover, the antiviral efficacy of STM2457 was confirmed in primary human hepatocytes, demonstrating an 80% reduction in infection at a 10 µM drug concentration. These findings were further validated by siRNA knockdown experiments, which demonstrated that the depletion of the m6A writer METTL3 and METTL16 by siRNAs decreased HEV infections by 41% and 12%, respectively. In contrast, depletion of writer proteins METTL14 and WTAP as well as the reader protein YTHDF3 and the eraser FTO increased HEV infectivity. In conclusion, our study identified m6A methylation as a component of HEV biology and suggests that it represents a promising target for antiviral interventions.

The human angiotensin-converting enzyme 2 (hACE2) serves as the primary receptor for SARS-CoV-2 entry via its interaction with the viral spike (S) protein. While certain ACE2 alleles are associated with enhanced binding affinity to the spike receptor-binding domain (RBD), the broader impact of ACE2 polymorphisms on infection susceptibility remains unclear. Building on previous findings that residues G496 and F497 in the spike protein and D355 and Y41 in ACE2 are critical for RBD–ACE2 binding, we employed computational saturation mutagenesis to systematically evaluate the effects of all possible ACE2 missense mutations on spike stability and binding. Our computational screening of ACE2 polymorphisms identified mutations at glycine residues (G268, G399, G405, G486, and G561) as having the most destabilizing effects on protein stability. We identified key residues that modulate binding affinity and pinpointed six ACE2 regions (residues 19–49, 65–102, 320–333, 348–359, 378–395, and 552–563) as candidate neutralizing peptides. Experimental assays confirmed that several of these peptides bind the spike protein and inhibit SARS-CoV-2 infection in ACE2-expressing cells. Comparative structural analysis between Wuhan and Omicron S1–ACE2 complexes revealed conserved binding patterns, indicating that these peptides may retain inhibitory potential across SARS-CoV-2 variants. These findings underscore the importance of considering host genetic diversity in understanding SARS-CoV-2 susceptibility and highlight the potential for designing ACE2-derived neutralizing peptides to block viral entry.

Antimicrobial resistance (AMR) is a complex problem that threatens human and animal health and overall global security. In 2019, AMR was listed by the World Health Organization (WHO) as one of the top 10 dangers in the World. AMR is on the rise, and if current trends continue unchecked, up to 10 million fatalities per year could occur by 2050. To address the problems caused by AMR, international organizations, including the World Health Organization, the FAO, and other relevant organizations, have suggested the One Health Approach, which is applicable in multiple sectors. However, despite growing importance, there remains a limited understanding of how One Health principles have been operationalized and integrated within AMR research worldwide. Additionally, there are inadequate reviews on multidisciplinary fields and limited quantitative reviews. This study uses a systematic and bibliometric review to address this gap by evaluating the conceptual structure, performance, and further examining how the One Health Approach is operationalized within AMR research. Relying on the PRISMA protocol, this study analyzes ninety-six (96) papers extracted from the Scopus and PubMed databases from 2010 to 2025 using Vosviewer and R-biblioshiny. The results show that the United Kingdom, the United States, and India are the top three (3) prominent countries in AMR research under the One Health Approach. Ballash, G.A., is the most influential author. Furthermore, the emergent themes of AMR research under the One Health Approach are as follows: One Health policy dimensions and public health, microbiology and mechanisms of bacterial resistance, and pharmacological and clinical studies. The results from this systematic review show that the integration of the One Health Approach has progressed from concept to practice, particularly in the health, veterinary, and environmental sectors. Hence, the study recommends an increase in investment, research, and innovation across all sectors, especially sectors such as agriculture that have received less attention.

Introduction

Coronaviruses (CoVs) pose a major risk to human health, exemplified by the emergence of SARS-CoV-2 and COVID-19. During infection, viruses depend on host factors for replication, while cellular mechanisms act to restrict infection. Non-coding RNAs (ncRNAs) are increasingly recognized as key regulators of cellular processes. Studying host long ncRNAs (lncRNAs) during infection may reveal novel virus–host interactions and potential therapeutic targets.

Objectives

We aim to elucidate the role of host lncRNAs during CoV infection to enable the identification of new targets for pharmaceutical intervention. In more detail, this project aims to identify, functionally validate and characterize lncRNAs to dissect their role during CoV replication.

Materials and methods

To identify lncRNAs involved in host cellular responses, human airway epithelial cells (hAEC) were either challenged with SARS-CoV-2 or treated with IFN-α or IFN-λ. We performed a microarray screen and conducted bioinformatic analysis to identify dysregulated lncRNAs following treatment. To validate the identified lncRNAs, their expression was altered using siRNA-mediated knockdown. Cells were then challenged with CoVs to assess the impact on viral replication.

Results

Bioinformatic analysis revealed differential expression of lncRNAs between the different treatment regimens. IFN-α treatment induced significant changes in a large number of lncRNAs, whereas IFN-λ resulted in significantly less dysregulation. Following CoV infection, few lncRNAs were differentially expressed at 16hpi, with GO analysis linking down-regulated lncRNAs to autophagy. At 72hpi, an increased number of lncRNAs was dysregulated, with pathways associated with the immune response. First results point towards reduced viral replication upon knockdown of specific lncRNAs, suggesting functional relevance.

Conclusions

We were able to demonstrate altered lncRNA expression upon IFN stimulation and viral infection. GO analysis implicated a potential role of lncRNAs during CoV immune control. Future studies will functionally characterize specific lncRNAs during virus replication and immune regulation, offering new therapeutic strategies.

Human norovirus (HuNoV) is the leading cause of non-bacterial acute gastroenteritis, imposing a substantial global health and socioeconomic burden. Although commensal bacteria have been shown to influence viral infectivity by facilitating attachment and modulating host immune signalling, their role in HuNoV recognition at the intestinal interface remains poorly understood. This study investigated whether commensal Escherichia coli (E. coli) influences Toll-like receptor (TLR)-mediated recognition and transcriptional activation during HuNoV exposure using a microfold (M) cells 3D Alcart model. The set up was established by co-culturing Caco-2 colonic epithelial cells with Raji B cells within alginate hydrogel beads as a recapitulation of the intestinal environment. The model was inoculated with HuNoV virus-like particles (VLPs) in the presence or absence of E. coli, and the TLR3 and TLR7 levels of expression were analysed by flow cytometry. Expression levels of transcription factor IRF3, IRF7, NF-κB1, and NF-κB2 genes were quantified with Real-Time PCR at 24 and 48 hours post-inoculation to assess early immune signalling. Flow cytometry results showed that at 24 hours post-inoculation, co-exposure to HuNoV VLPs and E. coli enhanced TLR3 expression but transiently suppressed TLR7, whereas by 48 hours, both receptors were markedly upregulated, indicating a time-dependent immune activation. Transcription factor gene analysis revealed early but short-lived activation of IRF3, IRF7, NF-κB1, and NF-κB2, suggesting that TLR activation precedes downstream gene induction. By 48 hours, TLR3 and TLR7 remained elevated while transcription factor expression returned to baseline, indicating transient immune activation. These findings demonstrate that commensal bacteria enhanced TLR3-mediated recognition of HuNoV while transiently suppressing TLR7 signalling in the M cells 3D Alcart model. Evidently, the M cells 3D Alcart presents a physiologically relevant platform for HuNoV pathobiology study, particularly on the influence of commensal bacteria towards antiviral responses while limiting excessive inflammation.

INTRODUCTION: Epstein–Barr virus (EBV)+ Diffuse Large B Cell Lymphoma (DLBCL) is a new entity confirmed by WHO. EBV may act as an alternative or complementary mechanism to the genetic alterations involved in DLBCL. Since sensitive methods have described traces of EBV infection in cases originally considered negative, the role of EBV in DLBCL pathogenesis is still under discussion.

AIM: This study aims to analyze the genetic alterations, classify them according to their pathogenicity, and correlate these with the presence of the virus and its traces.

METHODS: EBV+DLBCL was defined by EBER in situ hybridization, with 20% of EBERs+ tumor cells as the cut-off. Viral traces were analyzed using a ViewRNA assay to detect LMP1 and EBNA2 transcripts. Genetic variants were evaluated using a custom next-generation sequencing panel, focusing on pathogenic variants and pathway enrichment. Comparative analyses were conducted among EBV+ DLBCL, EBV- DLBCL with and without traces.

RESULTS: NGS analysis identified pathogenic variants mainly in ATM, TP53, PTEN, ARID1B, and KMT2A genes across all groups, suggesting shared mechanisms of DNA repair dysfunction and cell cycle regulation. No significant association was found between the presence or traces of EBV and the frequency of pathogenic variants when they were analyzed as a whole. However, EBV+ DLBCL exhibited unique alterations in BTK, HAX1, PAFAH1B1, and NAGLU genes, which are implicated in immune regulation, apoptosis, and mitochondrial dynamics, while 75% of EBV+ DLBCL displayed variations in the C11orf65 gene. NOTCH pathway variants were exclusively enriched in EBV+ DLBCL.

CONCLUSION: EBV and its traces do not impact pathogenic variants in DLBCL, suggesting an epigenetic or immunomodulatory role in lymphomagenesis. Frequent mutations in ATM, TP53, and PTEN across all groups highlight genomic instability's role, while genetic alterations in specific genes and the NOTCH pathway in EBV+ DLBCL suggest virus-specific mechanisms (not mediated by traces) that may promote tumorigenesis.

Background: Healthcare workers (HCWs) have been at the frontline of the COVID-19 pandemic, particularly in low-resource settings such as Africa, where protective measures were often limited. Understanding the extent of SARS-CoV-2 exposure among HCWs is crucial for guiding future infection control and vaccination strategies.

Objective: This systematic review and meta-analysis aimed to estimate the overall seroprevalence of anti–SARS-CoV-2 antibodies among HCWs in Africa and to explore variations by age, sex, geographic region, study design, and study period.

Methods: Following PRISMA guidelines, we searched PubMed and Cochrane databases for studies published between January 2020 and May 2025. Thirty-three eligible studies were included. Data were extracted on study characteristics, serological methods, and prevalence estimates. Pooled prevalence was calculated using random-effects models.

Results: The overall pooled seroprevalence of anti–SARS-CoV-2 antibodies among African HCWs was 23% (95% CI: 16–31%), indicating a high level of exposure. Considerable heterogeneity was observed (I² = 99.3%, p < 0.0001), with substantial variation across countries. The highest prevalence rates were reported in Mali (63%), Ethiopia (56%), and Cameroon (54%), whereas lower rates were found in South Africa (4%) and Mauritania (2%). Most studies used rapid antibody tests (IgG/IgM), and sampling was predominantly convenience-based. A non-significant trend toward higher seroprevalence among HCWs under 40 years of age and among women was observed.

Conclusions: African healthcare workers exhibited a high seroprevalence of SARS-CoV-2 antibodies, more than twice that reported in other global regions during the same period. This reflects increased occupational vulnerability due to limited personal protective equipment, weak infection prevention systems, and high community transmission in the absence of vaccination. Strengthening longitudinal research capacity, standardizing serological protocols, and enhancing HCW protection are essential to mitigate future infectious disease crises.

Introduction

Hepatitis E virus (HEV) infections represent the leading cause of acute viral hepatitis globally, accounting for over 20 million cases annually. Despite their prevalence, our understanding of the HEV replication cycle and its interactions with the host cell remains limited. Recent studies have highlighted the involvement of peroxisomes—organelles essential for lipid and fatty acid metabolism, detoxification processes, and innate immune responses—in the life cycles of various viruses. This study aims to elucidate the role of peroxisomes in the HEV replication cycle and to determine their significance as host organelles in HEV infection.

Material and Methods

HEK T-REx293 cells with individual knockouts of eleven peroxins (PEX) essential for peroxisome biogenesis were generated. Cells were infected with HEV genotype 3 (Kernow-C1 p6), and infection was assessed using focus forming assays. Additionally, a Gaussia luciferase reporter replicon was used to evaluate replication in the knockout lines. Wild-type cells served as controls.

Results

HEV infection assays showed a two- to three-fold increase in focus forming units in PEX1- and PEX5-knockout cells compared to wild-type controls, indicating enhanced viral replication. This observation was corroborated by the Gaussia luciferase-based replication assay, which demonstrated a similar increase in reporter activity in PEX1-KO and PEX5-KO cells. Conversely, knockout of PEX3, PEX7, and PEX10 resulted in a marked reduction in HEV replication capacity.

Conclusion

Collectively, these findings suggest that peroxisomes play a significant role in the HEV replication cycle. Notably, the absence of PEX1 and PEX5, which are essential for the import of matrix proteins into peroxisomes, appears to facilitate HEV replication, whereas disruption of other peroxins impairs viral propagation. These results provide novel insights into the interplay between peroxisomal function and HEV biology and highlight specific peroxins as potential modulators of HEV infection.

Systemic lupus erythematosus (SLE) is a complex autoimmune disease that continues to raise major questions regarding its origin and progression. In recent years, Epstein–Barr virus (EBV) has gained particular attention as a potential trigger and modulator of the immune response in these patients. This study aimed to better understand this relationship by exploring how viral infection and the immunological environment of SLE influence gene regulation and immune cell activity.

When comparing SLE patients with healthy controls, we found that most cases exhibited active disease, with a high frequency of renal involvement. In these patients, the expression of key genes such as TNF-α and IFN-γ was reduced, whereas that of IL-10 was increased and inversely correlated with TNF-α, indicating a disrupted balance in the inflammatory response. Moreover, most patients showed evidence of active EBV infection, associated with the overexpression of LMP1, a viral protein capable of promoting the survival of autoreactive B cells and altering critical immune signaling pathways.

Another relevant finding was the activation of endogenous retroviruses (HERV-E). In heterologous plasma assays, we also confirmed that the lupus environment can directly modify the behavior of healthy immunocompetent cells, particularly by promoting changes in IL-6. Finally, we identified cross-reactivity between EBNA1 and Ro60, supporting the hypothesis of molecular mimicry as one of the mechanisms underlying the loss of immune tolerance.

Taken together, this study provides an integrated view of the role of EBV, endogenous retroviruses, and immune dysregulation in the pathogenesis of SLE. Beyond expanding the biological understanding of the disease, these findings suggest potential biomarkers and therapeutic pathways that may contribute to more precise and personalized management of lupus in the future.

Background: Epstein–Barr virus (EBV) infection alternates between latency and reactivation, accompanied by EBV DNA shedding that can trigger proinflammatory immune responses implicated in autoimmune diseases such as rheumatoid arthritis (RA). Our group previously showed that EBV DNA increases the production of IL-17A, a proinflammatory cytokine, in mice through endosomal toll-like receptor (TLR) signaling. We also demonstrated that EBV DNA enhances the incidence and severity of arthritis in a collagen-induced arthritis (CIA) model using C57BL/6J mice. This study aimed to determine the effect of endosomal TLR inhibition on EBV DNA-exacerbated arthritis in this model.

Methods: Female C57BL/6J mice were injected with type II chicken collagen and treated with EBV DNA alone or combined with inhibitors of TLR3, TLR7, or TLR9. Control groups received each inhibitor alone. Arthritis severity was evaluated by paw thickness, clinical scoring, and grip strength. Histological analyses of footpads, ankle joints, and colons were performed, and the frequency of IL-17A, IFN-γ, and FOXP3 co-expressing immune cells in ankle joints was assessed using immunofluorescence and confocal microscopy.

Results: Mice treated with EBV DNA, collagen, and an endosomal TLR inhibitor exhibited significant reductions in paw thickness, arthritis scores, and histological damage, alongside improved grip strength, compared to those treated with EBV DNA and collagen alone. Among the inhibitors, TLR9 blockade produced the most pronounced reduction in clinical and histological severity. Inhibitor treatment also decreased the number of IL-17A⁺IFN-γ⁺FOXP3⁺ cells in ankle tissues.

Conclusions: Inhibition of TLR3, TLR7, or TLR9 attenuates EBV DNA-exacerbated arthritis in a CIA mouse model, with TLR9 inhibition showing the strongest therapeutic benefit. These findings highlight endosomal TLRs, particularly TLR9, as promising therapeutic targets for RA management in EBV-infected individuals.