Epidemiological characteristics and viral molecular evolution analysis of severe fever with thrombocytopenia syndrome in Shandong Province, China.

Oral administration of IFN-γrel elicits antiviral immunity and enhances protective efficacy against LMBV in largemouth bass (Micropterus salmoides).

Molecular characterization and functional analysis of IL-19 like in large yellow croaker (Larimichthys crocea).

We have generated two reporter plasmids that either monitor expression of human papillomavirus type 16 (HPV16) E2 mRNAs using splice site SA2709 or HPV16 E1C mRNAs utilizing splice site SA2582. In plasmid pE2sLuc, the entire E2 open reading frame was replaced by the secreted luciferase (sLuc) reporter gene and in the pE1nLuc plasmid the E1/E1C open reading frame was fused in frame with the nanoluciferase (nLuc) reporter gene. We demonstrate that these reporter plasmids can be used to investigate cis-acting RNA elements and trans-acting factor that regulate splicing to the E2 splice site SA2709 and the alternative E1C- splice site SA2582. Furthermore, we generated a stable C33A cell line carrying HPV16 reporter plasmid pE1nLuc. Using this cell line, we showed that splicing to the HPV16 E2 3'-splice site SA2709 could be perturbed by over expression of SR-proteins or by incubation with a pan-Akt kinase inhibitor. These proteins and substances shifted splicing from E2 splice site SA2709 to the upstream HPV16 3'-splice site SA2582 and enhanced nLuc production, thereby demonstrating the utility of the pE1nLuc cell line as a bioassay for HPV16 E2 mRNA splicing.

Molecular characterization of peroxiredoxin 5 from Mytilus coruscus and its roles in antioxidant defense and immune regulation.

Identification and expression analysis of scavenger receptor SCARA4 in Nibea albiflora.

Finely-tuned regulator of DNA repair: FBXW7-185aa may modulate glioblastoma radiosensitization and the tumor immune microenvironment.

Genome characterization and evolutionary analysis of a novel Anativirus from a wild Muscovy duck in Guangdong, China.

Molecular characterization and functional analysis of EBF1 reveals its critical role in B-cell immunity in Nile tilapia.

Escherichia coli is a major cause of hospital-acquired infections in China, including urinary tract, bloodstream, and intestinal infections. Given the rising prevalence of antibiotic-resistant E. coli, phages are increasingly regarded as promising alternatives to conventional antibiotics. Henuyfy11N was isolated using the double-layer agar method and characterized via transmission electron microscopy in this study. Biological assays included stability under varying pH and temperature, UV sensitivity, host range, optimal multiplicity of infection, adsorption rate, and one-step growth curve. In vitro lytic activity against extended-spectrum β-lactamase (ESBL)-producing E. coli and biofilm eradication capacity was assessed. Whole-genome sequencing enabled phylogenetic, synteny (the analysis of conserved blocks of genetic sequence between different genomes), and functional annotation analyses. In vivo, the therapeutic efficacy was evaluated in a mouse infection model. Phage Henuyfy11N has not yet been classified. It demonstrated high lytic activity, a short latent period, and a burst size of 57.1 PFU/cell. The phage remained stable across a broad pH range (3-11) and temperatures up to 70℃. Its circular double-stranded DNA genome (41,103 bp, G + C% 50.88) contains 54 open reading frames, with no tRNA, virulence, or antibiotic resistance genes. Genomic and phylogenetic analyses revealed close relatedness to phage BUCT789. Henuyfy11N effectively lysed ESBL-producing E. coli, disrupted biofilms, and significantly improved survival in the mouse infection model. Henuyfy11N shows high host specificity, efficient lytic activity, rapid replication, and a safe genomic profile, demonstrating some potential as a therapeutic agent against ESBL-producing E. coli infections.IMPORTANCEThe widespread use of antibiotics has led to increasing antibiotic resistance, which is a growing global health concern. Therefore, the development of novel antimicrobial therapy that can cure drug-resistant bacteria-induced infections is imperative. Phages are of increasing interest as natural enemies of bacteria, with clear advantages in antibacterial applications. In this study, by using extended-spectrum β-lactamase (ESBL)-producing Escherichia coli 2025011N as a host, we successfully isolated and purified Escherichia phage Henuyfy11N and conducted a series of experiments to verify its genomic character and biological character. Our findings revealed that the phage exhibited excellent tolerance to a broad spectrum of pH and wide temperature range. Phage Henuyfy11N was effective in disrupting mature biofilm, and no genes for virulence, lysogenic, integrase, or AMRs were found in the genome. Besides, Henuyfy11N showed promising antibacterial effects in vivo and in vitro, indicating potential as a therapeutic agent against ESBL-producing E. coli infections.

Millettia pinnata (M. pinnata) is a medicinally important plant containing many phytoconstituents capable of curing various diseases. Flavonol synthase (FLS, EC 1.14.20.6) has a significant role in flavonoid biosynthesis like kaempferol having various pharmacological properties. The present study aimed to characterize the FLS gene and its encoded protein (MpFLS) in M. pinnata comprehensively. FLS gene was characterized by isolating RNA from M. pinnata leaves, cDNA synthesis, PCR-based amplification of the desired FLS gene, sequencing of the PCR amplified product and sequence analysis followed by detailed protein-protein interaction and protein flexibility analysis. PCR product sequencing and analysis revealed that the full-length cDNA of M. pinnata FLS contains a 952-base pair long open reading frame (ORF) encoding 234 amino acids that has been submitted to NCBI (OM469017). The NCBI blastp analysis of 234 amino acid sequences of FLS shows 81.9% similarity to Vigna unguilata, whereas blastn analysis exhibited 83.31% identity to Clitoria ternatea L. that belongs to the 2OG-FeII Oxy superfamily. In silico characterization of primary, secondary, and tertiary structure, docking of MpFLS with dihydrokaempferol showed a strong binding (-10.0kcalmol⁻1) stabilized by hydrogen bonds (Gly54, Glu256) and hydrophobic/π-π interactions, highlighting a specific and stable enzyme-substrate complex. STRING based protein-protein interaction analysis and detailed ProtScale and MEDUSA based protein flexibility analysis of FLS protein revealed that FLS it is highly stable and hydrophilic. M. pinnata FLS gene have 952-base pair long ORF while encoding FLS protein of 234 amino acids long. FLS belong to 2-oxoglutarate/Fe(II)-dependent oxygenase superfamily that utilize molecular oxygen and 2-oxoglutarate to oxidize various substrates. Molecular docking results indicate that MpFLS binds dihydrokaempferol with high specificity and stability, supporting its key role in kaempferol biosynthesis in Pongamia pinnata. Present study provides systematic and comprehensive analysis on FLS gene and its encoded FLS protein in M. pinnata revealing that FLS protein is highly stable and hydrophilic in nature.

Toxoplasma gondii is a globally prevalent protozoan parasite capable of establishing lifelong infections in its host. While acute infection is often asymptomatic, reactivation of latent bradyzoites can cause severe disease, particularly in immunocompromised individuals. Current therapies are ineffective against chronic infection, underscoring critical gaps in our understanding of bradyzoite biology and the molecular mechanisms governing stage conversion. Recent studies have identified translational control as a central regulator of T. gondii differentiation. This review highlights the roles of canonical translation initiation factors (eIF2α, eIF1.2, and eIF4E1), RNA-binding proteins (RBPs; BFD2/ROCY1, Alba1, and Alba2), and RNA modifications (with pseudouridylation representing the best-characterized modification currently linked to differentiation), as well as alternative splicing and non-coding RNAs in shaping stage-specific translational programs. This review further discusses underexplored mechanisms, including non-canonical initiation pathways, upstream open reading frames, transcript-level RNA modifications, ribosome heterogeneity and rRNA modifications, elongation and termination control, uncharacterized RBPs, and post-translational modifications of translation factors, that may coordinate proteome remodeling during differentiation. Together, established translational regulators and these emerging pathways highlight translational control as a central driver of parasite persistence and a promising therapeutic target for chronic toxoplasmosis.

ABSTRACT Influenza A viruses (IAVs) lacking the nonstructural protein 1 (NS1) gene (IAV-ΔNS1) are attenuated both in vitro and in vivo, yet retain replicative capacity in interferon signaling-deficient systems. To date, several efforts have been pursued to expand its functionality by incorporating transgenes in lieu of the NS1 gene. However, such vectors require complex segment designs, retain residual NS1 peptide stretches, or display suboptimal cytokine secretion, largely due to the constraints related to the partial nuclear export protein (NEP) open reading frame (ORF) upstream of the transgene integration site. Here, we describe a novel segment 8 design that can accommodate a transgene, where the complete NEP ORF is positioned downstream of the splicing acceptor site, thereby mitigating previous genetic constraints. Using this strategy, we successfully generated a recombinant IAV-ΔNS1 virus carrying the human interleukin 2 (IL-2) gene, inducing robust secretion of bioactive IL-2 upon infection. The vector demonstrated high replicative capacity, achieving titers comparable to wild-type viruses in MDCK cells expressing NS1 protein, and exhibited genetic stability over 10 successive passages. Importantly, infection induced notable viral antigen expression and high levels of bioactive IL-2 secretion in interferon-competent systems, such as MDCK, A549, and HEK293T cells in vitro and in mice upon pulmonary delivery in vivo . Furthermore, the transgene was easily swapped with a diverse array of genes encoding human IL-15, nanoluciferase, or fluorescent proteins such as miniSOG and miRFP670nano, highlighting the versatility of the platform. IMPORTANCE IAV-ΔNS1 vectors are promising vaccine platforms that elicit strong immune responses with a good safety profile. However, integration of immune-stimulatory cytokines into vector design to boost immunogenicity has been technically challenging. In this study, we developed a genetically re-engineered segment 8 design that overcomes prior limitations due to design complexity or vector efficiency, enabling high-level expression of the transgene, i.e., human interleukin 2 and other diverse proteins. The platform retains strong replicative capacity in permissive systems and remains genetically stable, making it suitable for scalable vaccine or therapeutic development. By improving both the flexibility and functionality of IAV-ΔNS1 vectors, our work advances the utility of influenza A viruses as customizable tools for vaccine delivery, immune modulation, and therapeutic applications.

Pseudomonas aeruginosa is an important cause of infections in hospitalized paediatric patients, which can prolong hospital stays, increase medical care costs, and increase morbidity and mortality. The aim of this study was to describe the molecular characteristics of carbapenem-resistant P. aeruginosa (CR-PA) isolates obtained from paediatric patients, along with their clinical data, infection type, treatment, and outcome. From January 2018 to March 2020, 65 P. aeruginosa isolates were prospectively collected, and carbapenemases were detected. The genes mexR, nalC, nalD, and oprD were amplified, sequenced, and analysed. The sequence types (STs) were determined by Multilocus sequence typing. Clinical information on the origin, evolution, treatment, and outcome of the infections was collected. Most isolates (81.6%, n = 53) were carbapenemase-negative; 18.4% (n = 12) were carbapenemase producers, with IMP being the most common (7/12). In the sequences of the mexR, nalC and nalD genes, 25% (n = 16), 8.5%(n = 6), and 10.6%(n = 7), respectively, presented modifications in the open reading frame, and stop codons were observed in 17.3% of mexR and 2.1% of nalD sequences. Premature stop codons were present in 70.6% of the oprD sequences, and frameshift mutations were observed in 75.5%. The isolates were distributed among 35 ST; ST348 (16.9%, n = 11), ST309 (12.3%, n = 8), and ST389 (10.8%, n = 7) were the most frequent. High-risk clones (HRC) including ST358, ST309, ST389, ST395, ST234 and ST11 were detected. A total of 92% (n = 60) of patients with CR-PA had underlying health conditions, the most frequent being congenital disorders. The most common empirical therapy was a combination of meropenem and an aminoglycoside or colistin (52.6%, n = 34), while among definitive treatments, monotherapy was used in 67.7% of patients, and 29.2% received a two-drug combination. The complications were associated with mortality (RR = 2.5, p < 0.0001). A total of 89.2% of infections were hospital-acquired. Modification of the porin OprD was the main mechanism of carbapenem resistance among P. aeruginosa isolates; however, carbapenemase-producing strains were observed, mainly from the imipenemase (IMP) family. 60% of the isolates belonged to STs, considered HRC. Of the clinical variables analysed, only the presentation of complications was associated with mortality.

An adaptor protein CRK mediates antiviral immunity against white spot syndrome virus via JAK/STAT signaling pathway in Cherax quadricarinatus.

A macrophage mannose receptor 1 with dual CTLDs played a crucial role in defense against Ecytonucleospora hepatopenaei in Litopenaeus vannamei.

Reversible conformational dynamics of membrane proteins are essential for intracellular signaling, but no method enables their irreversible arrest in living cells. Here, we developed a genetically encoded proximity-based lockdown enzyme derived from an engineered transglutaminase catalytic core (TGC) that covalently crosslinks membrane proteins. By fusing TGC to the endoplasmic reticulum (ER)-resident microprotein ALN encoded by a short open reading frame (sORF), we created an organelle-specific module that selectively catalyzes covalent crosslinking within the SERCA Ca2+ pump, strongly suppressing its ATP-dependent pump activity and arresting ER Ca2+ signaling. This engineered lockdown enzyme remodels ER membrane protein architecture and restricts conformational dynamics, providing a versatile platform for long-term covalent control of intracellular signaling and a foundation for future therapeutic cellular applications. Impact statement Our proximity-based lockdown enzyme, engineered from microbial transglutaminase, provides a new strategy to covalently arrest the conformational states of organelle-resident membrane proteins in living cells, enabling long-term control of intracellular signaling and establishing a foundation for next-generation cellular therapeutics.

OfERF106 protein binds to the promoter region of the ethylene biosynthesis geneOfACS2and activates its expression, thereby accelerating the aging process of Osmanthus fragrans petals. Osmanthus fragrans is a multipurpose ornamental plant that combines landscaping utility, aesthetic appeal, and aromatic fragrance, granting it considerable economic and horticultural value. However, its value is substantially constrained by the short overall flowering period and the brief longevity of individual petals. The key regulatory genes, their functions, and the molecular mechanisms underlying petal senescence in O. fragrans remain largely unknown. Here, we identified a critical gene, OfERF106, from transcriptomic data. The expression level of OfERF106 peaked at the flower fading stage and was significantly higher than that observed during the primary and full blooming stages. The full-length sequence of OfERF106 contains a 675bp open reading frame encoding 225 amino acids. Subcellular localization analysis confirmed that OfERF106 protein is localized in the nucleus. Overexpression of OfERF106 in Arabidopsis thaliana resulted in a flowering period shortened by 3-4days compared to the control, indicating its role in promoting petal senescence. The functional validation of OfERF106 through rapid overexpression and silencing in O. fragrans petals further supported this result. Moreover, the expression levels of two ethylene biosynthesis-related genes (OfACS1 and OfACS2) were also significantly altered. Further analysis using yeast one-hybrid (Y1H) and dual luciferase (DLR) assays demonstrated that OfERF106 protein binds to the promoter of OfACS2 and activates its expression. These results suggest that OfERF106 accelerates petal senescence in O. fragrans by enhancing OfACS2 expression, which in turn promotes ethylene biosynthesis. Our study provides preliminary insights into the molecular mechanism regulating petal senescence in O. fragrans and highlights the functional role of OfERF106 in this process.

Genomic characterization of a vancomycin-intermediate Staphylococcus aureus (VISA) small colony variant after long-term antibiotic therapy.

The Mycobacterium tuberculosis (Mtb) VapC4 endoribonuclease toxin exclusively cleaves and inactivates tRNACys, which leads to extensive ribosome stalling at Cys codons. Serendipitously, the precise position of stalled ribosomes is revealed within our 5' RNA-seq datasets used to identify and validate the tRNA target of the toxin, precluding the need for Ribo-seq. Here we show how mapping of stalled ribosomes can be harnessed as an innovative tool for reliable detection of new Cys-containing Mtb open reading frames (ORFs). Using proteogenomics we unmasked 96 unannotated ORFs; of which 54% are small ORFs ≤50 amino acids. We validated 69% of the 96 ORFs by mass spectrometry, including four whose spectra was matched to synthetic controls Also, 25% of these unannotated ORFs were identified by previously published Ribo-RET. Some of the 96 ORFs are Cys-responsive attenuators or encode stable Cys-containing proteins that map immediately before, or within, genes in the opposite, or same, orientation. These ORF sequences can also reveal functional clues, e.g. zinc-binding motifs or encode novel EsxB-like proteins. Our findings demonstrate that toxin-mediated ribosome stalling can serve as a robust genome annotation tool that is applicable to mycobacteria and other bacteria, with unique advantages that complement existing genome annotation methods.