Journal: Emerging Microbes & Infections

Article Title: Lactoferrin blocks orthopoxvirus entry via heparan sulphate and regulates host antiviral pathways

doi: 10.1080/22221751.2026.2631205

Figure Lengend Snippet: Lactoferrin inhibits orthopoxvirus infection by blocking viral entry through heparan sulphate binding. (A) Time-of-addition kinetics of 2.5 mg/ml bLf against vvTT infection (MOI = 0.1). Viral loads quantified at 14 hpi by Western blot and RT-qPCR. Data = mean ± SD ( n = 3). **** p < 0.0001 (one-way ANOVA with Dunnett's test). (B) Representative immunofluorescence images of vvTT-infected Vero E6 cells treated with 2.5 mg/ml bLf at indicated time points. Viral A27L protein (green), nuclei (DAPI, blue). Scale bars = 100 μm. (C) The inhibitory activity of lactoferrin at concentrations of 2.5, 1.25, and 0.625 mg/ml during the entry stage was quantified using RT-qPCR. Data = mean ± SD ( n = 3), **** p < 0.0001 (one-way ANOVA with Dunnett's test). (D) Post-entry curve depicted the replication dynamics of vvTT (MOI = 0.01) in Vero E6 cells after 2.5 mg/ml lactoferrin treatment. Tecovirimat at a concentration of 100 nM served as a positive control. Viral copies were quantified using a standard plasmid and analysed by absolute quantification methods. Data = mean ± SD ( n = 3). (E) Heparin competition assay. Lactoferrin (250 μg/ml) pre-incubated with heparin (HS) at indicated ratios during vvTT infection (MOI = 0.1). Viral loads quantified by RT-qPCR at 24 hpi. Data = mean ± SD ( n = 3). (F) Differential scanning fluorimetry showing direct lactoferrin-heparan sulphate binding. Thermal shift (Tm) of bLf (100 μg/ml) with increasing HS concentrations (0.02-200 μg/ml). Data = mean ± SD ( n = 3). (G) Sodium chlorate pre-treatment depletes cell-surface HSPGs and inhibit vvTT (MOI = 0.1) infection. Viral yields quantified by RT-qPCR at 24 hpi. Data = mean ± SD ( n = 3).

Article Snippet: Antiviral reference compounds brincidofovir (Cat. TQ0095, TargetMol) and tecovirimat (Cat. V3886, InvivoChem) were used for combination therapy studies.

Techniques: Infection, Blocking Assay, Binding Assay, Western Blot, Quantitative RT-PCR, Immunofluorescence, Activity Assay, Concentration Assay, Positive Control, Plasmid Preparation, Quantitative Proteomics, Competitive Binding Assay, Incubation

Journal: Emerging Microbes & Infections

Article Title: Lactoferrin blocks orthopoxvirus entry via heparan sulphate and regulates host antiviral pathways

doi: 10.1080/22221751.2026.2631205

Figure Lengend Snippet: Bovine lactoferrin exhibits therapeutic efficacy against VACV infection in vivo . (A) Experimental design for prophylactic bLf treatment. (B) Quantification of viral load in lung tissues. Viral genome copies were quantified as genomes per gram of tissue (left), while virus titres were calculated as PFU per gram of tissue (right). The data analysis performed using GraphPad Prism version 8.0. Data = mean ± SD ( n = 4), * p < 0.05, ** p < 0.01 (two-tailed t -test). (C) Quantitative analysis of infectious viral particles in the lung with 100 mg/kg lactoferrin, 50 mg/kg tecovirimat and PBS treatment based on plaque assay. (D) Weight loss dynamics and survival rates monitored daily for 7 days and clinical and pathological scoring of mice. (E) H&E staining and IHC analysis of the lung sections. The sections exhibited diffuse degeneration and necrosis of the epithelial lining (black arrows), along with haemorrhage, edema (red arrows), and fibrin exudation into the surrounding alveoli (blue arrows). IHC analysis conducted using a human anti-VACV A27L monoclonal antibody (brown signal). Mock was the blank control group without virus infection. The slides were digitized using a Pannoramic MIDI histoscanner (3DHISTECH), and the images analysed with CaseViewer software. Scale bars, each representing 50 micrometers, are provided for reference in each image.

Article Snippet: Antiviral reference compounds brincidofovir (Cat. TQ0095, TargetMol) and tecovirimat (Cat. V3886, InvivoChem) were used for combination therapy studies.

Techniques: Drug discovery, Infection, In Vivo, Virus, Two Tailed Test, Plaque Assay, Staining, Control, Software

Journal: Viruses

Article Title: Establishment of a Dual-Reporter Minigenome System for Respiratory Syncytial Virus

doi: 10.3390/v18030304

Figure Lengend Snippet: Mini-NLuc-sfGFP for small-molecule evaluation. ( A ) The experimental protocol involved transfecting the Mini-NLuc-sfGFP minigenome into BSR-T7/5 cells, followed by treatment with a range of small molecules. The inhibitory effects were initially assessed by quantifying the fluorescence intensity or NLuc activity, which led to the selection of promising candidates for further investigation. Created in BioRender. Wu, C. (2026) https://BioRender.com/8qct7m8 . ( B ) AVG-233 reduced the BSR-T7/5 cell viability in a concentration-dependent manner (OD 450 ). ( C ) Similarly, RSV L-protein-IN-4 reduced the BSR-T7/5 cell viability in a concentration-dependent manner (OD 450 ). ( D ) AVG-233 demonstrated a dose-dependent inhibition of Mini-NLuc-sfGFP reporter gene expression, as indicated by the luciferase activity. ( E ) Similarly, RSV L-protein-IN-4 exhibited the dose-dependent inhibition of the Mini-NLuc-sfGFP reporter gene expression, also measured using the luciferase activity. Data are presented as the mean ± SD of three independent experiments, each performed in triplicate.

Article Snippet: Small-molecule inhibitors of RSV L polymerase, namely, AVG-233 (CAS no. 2151937-80-1) and RSV L-protein-IN-4 (CAS no. 851657-60-8), both sourced from Topscience Co. Ltd., China, were diluted in DMSO to create stock solutions at various concentrations.

Techniques: Fluorescence, Activity Assay, Selection, Concentration Assay, Inhibition, Gene Expression, Luciferase

Journal: Viruses

Article Title: Establishment of a Dual-Reporter Minigenome System for Respiratory Syncytial Virus

doi: 10.3390/v18030304

Figure Lengend Snippet: Validation of small-molecule inhibitory effects assessed by Mini-NLuc-sfGFP and confirmed in RSV A2. RSV A2 infection was conducted at a multiplicity of infection (MOI) of 0.1, utilizing a primary antibody of mouse anti-RSV F and a secondary antibody of goat anti-mouse FITC conjugate. ( A ) Immunofluorescence imaging showed that AVG-233 inhibited viral infection at 0, 0.01, 0.05, 0.25, 0.5, 1, 2, and 3 μM. ( B ) Viral genome copy numbers via RT-qPCR at equivalent concentrations. ( C ) Immunofluorescence imaging showed that RSV L-protein-IN-4 inhibited viral infection at 0, 0.01, 0.05, 0.25, 0.5, 1, 2, and 3 μM. ( D ) Viral genome copy numbers via RT-qPCR at equivalent concentrations. Scale bar: 400 μm. Data are presented as the mean ± SD of three independent experiments, each performed in triplicate.

Article Snippet: Small-molecule inhibitors of RSV L polymerase, namely, AVG-233 (CAS no. 2151937-80-1) and RSV L-protein-IN-4 (CAS no. 851657-60-8), both sourced from Topscience Co. Ltd., China, were diluted in DMSO to create stock solutions at various concentrations.

Techniques: Biomarker Discovery, Infection, Immunofluorescence, Imaging, Quantitative RT-PCR

Journal: Alzheimer's Research & Therapy

Article Title: Proof-of-concept study: APOE4 brain endothelial cells as a phenotypic compound screen

doi: 10.1186/s13195-026-01960-6

Figure Lengend Snippet: Compound treatment phase, mTOR inhibitors disrupt baseline TEER of APOE4 -brain endothelial cells. APOE4 -brain endothelial cells were treated for 24 h with 1 µM of 900 + compounds selected from the TargetMol Bioactive Library. A Capacitance was > 4nF for 11 compounds, 4 of which inhibit cell cycle pathways (CDK, purple dots) and 3 PI3K/Akt/mTOR (blue dots). B , C Relative change and percentage activity of compounds. We identified compounds that either increased (light green dots, 5 compounds) or decreased (brown dots, 34 compounds) baseline TEER. 22 were inhibitors of mTOR/PI3K (blue dots) including rapamycin (arrow). E - I The ability of mTOR activators (NV-5138, L-leucine, 3BDO, MHY1485) to modulate LPS-induced TEER disruption was evaluated. E , G Compound treatment phase. E. High concentrations of mTOR activators were either toxic (2 mM L-leucine; 200 µM 3BDO) or F. disrupted TEER (100 µM 3BDO). H , I LPS Phase. High concentrations of NV-5138 (10 µM) mitigated LPS-induced TEER disruption. Data analyzed by one-Way ANOVA/matched Mixed-effects model (REML) followed by Dunnet’s multiple comparisons test comparing a compound concentration to the control group. In the compound treatment phase ( F ), * p < 0.05 for a given compound concentration compared to vehicle. In the LPS treatment phase ( H ), * p < 0.05 for a given compound concentration + LPS compared to the vehicle plus LPS group. n = 3. Upper lines indicate the concentrations different from LPS control for each compound. All statistical analysis is provided in Additional File 2

Article Snippet: We then screened a subset of ~ 900 molecules from the TargetMol Bioactive Library and identified two main groups compounds.

Techniques: Activity Assay, Disruption, Concentration Assay, Control

Journal: bioRxiv

Article Title: Acquired resistance to the PRMT5 inhibitor confers collateral sensitivity to MEK inhibition in MTAP-null non-small cell lung cancer

doi: 10.64898/2026.04.16.719008

Figure Lengend Snippet: High-throughput drug screen and MEK inhibitor sensitivity in MRTX1719-resistant NSCLC cells. (A) Composition of the compound library used for drug screen, including SGC epigenetic compounds, FDA-approved oncology drugs, and TargetMol epigenetic inhibitors. (B) IC50 values of MRTX1719 and anisomycin in DMSO and MRTXR cells. Anisomycin was included as a nonselective control in the drug screen. (C) Dose-response curves of DMSO and MRTXR cells treated with the MEK inhibitor selumetinib. Data are presented as mean ± SD. (D) Synergy heatmaps of MRTX1719 and selumetinib in DMSO and MRTXR cells. Synergy mean scores were calculated using the Bliss model with the SynergyFinder+ tool.

Article Snippet: The screening library comprised 619 compounds, including SGC epigenetic compounds, TargetMol epigenetic inhibitors, and FDA-approved oncology drugs ( ).

Techniques: High Throughput Screening Assay, Drug discovery, Control