Search Results (14,329)

Hepatitis B virus (HBV) infection remains a major global public health burden, primarily due to the persistence of covalently closed circular DNA (cccDNA) in infected hepatocytes. Toll-like receptor 3 (TLR3) has been suggested to be a promising therapeutic target, as its activation stimulates innate immune responses that facilitate cccDNA clearance. In this study, a structure-based computational approach was employed to identify novel small-molecule TLR3 agonists. A human TLR3 ectodomain (ECD) homodimer model was constructed using homology modeling, revealing a critical cavity at the C-terminal interface, which is responsible for the ligand-binding-induced ECD dimerization. Subsequently, three compounds, ZINC3991624 (Vilanterol) and ZINC95564694 (Naloxegol) from the FDA-approved drug database along with TCM10752 from the Traditional Chinese Medicine (TCM) database, were identified as potential candidates through docking-based virtual screening, rigorous ADMET evaluation and molecular dynamics simulations. These candidates outperformed CU-CPT17e, the only known TLR3 agonist, by maintaining lower backbone RMSD values, preserving key inter-subunit distances and protein–protein interface area and sustaining C-terminal hydrogen-bonding networks. In conclusion, our study provides a structural basis for the further development of novel TLR3 agonists, offering a promising computational framework for future host-directed immunotherapeutics aimed at HBV cccDNA clearance. Full article

Sheep-associated malignant catarrhal fever (SA-MCF) is a frequently fatal, pantropic, lymphoproliferative disease caused by ovine gammaherpesvirus 2 (OvGHV2), with sheep serving as the asymptomatic reservoir host. Although the Rusa deer (Cervus timorensis) are known to be highly susceptible to OvGHV2 in other regions, cases have not been documented in this species from Brazil. This report describes the spontaneous occurrence of OvGHV2 infection in a captive Rusa deer from Southern Brazil and discusses possible impacts on wildlife. Three deer from the same property died suddenly without prior clinical signs. These animals had no known contact with sheep or goats, but were maintained in an epidemiological niche where OvGHV2 was detected in subclinically infected free-ranging wild boars (FRWBs) and cattle. One of these was submitted for routine post-mortem evaluations. Gross lesions included ulcerative and erosive lesions of the alimentary tract, pulmonary edema, and marked meningeal congestion. Histopathology revealed erosive and ulcerative esophagitis, abomasitis, omasitis, and rumenitis, with nonsuppurative meningoencephalitis, and vasculitis in several organs, consistent with SA-MCF. Molecular testing of brain tissue detected OvGHV2 DNA by qPCR. Furthermore, PCR assays amplified the partial fragments of the OvGHV2 tegument protein (ORF75) and the glycoprotein (gB) genes. Other neurological pathogens of ruminants including BoAHV5, BVDV, Listeria monocytogenes, Histophilus somni, and rabies virus were not detected. Additionally, phylogenetic analyses, based on ORF75 and gB genes, revealed that the nucleotide sequences derived from this study clustered with and had elevated sequence homology with the OvGHV2 reference strains and other strains derived from ruminants maintained in different geographical locations. The absence of contact with sheep, combined with prior evidence of subclinical OvGHV2 infection in FRWBs and cattle within the same epidemiological niche, suggests the involvement of alternative wildlife reservoirs or bridge hosts in viral transmission. Additionally, these results expand the host range of OvGHV2 in Brazil, highlight the vulnerability of exotic cervids to SA-MCF, and underscore the need for integrated wildlife–livestock surveillance strategies in regions where multiple susceptible species coexist. Full article

Avian metapneumovirus (aMPV) is an emerging viral pathogen causing many outbreaks in chickens all over the world. There are several subtypes of the MPV circulating in chickens in the US. The current circulating subtypes of the virus in the US chicken are A, B, and C. Despite the availability of some aMPV vaccines in the US, most of these vaccines are based on foreign strains, particularly European aMPV-A/B strains. Their protective efficacy against contemporary U.S. aMPV isolates has not been fully established. The main goal of this study is to integrate the most recent aMPV genome sequencing data and the machine learning tools to design a novel aMPV. The machine learning tools such as epitope mapping, molecular docking, and immune simulation were used to design the multiepitope DNA vaccine based on the top-ranked epitopes of two major surface proteins of the virus (F and G). The top-ranked seventeen epitopes representing B cells, CD4 and CD8 epitopes were linked using linkers, with IL-18 added as an adjuvant. The selected epitopes showed high antigenicity, no toxicity and no allergenicity values among the screened epitopes. The molecular docking analysis of the designed vaccine construct showed a high binding affinity to the MHC class I and II epitopes to chicken alleles. The immune simulation analysis of this vaccine provides an in silico proof of concept assessment through modeling of the vaccine potential to elicit humoral and cell-mediated immunity. Further functional studies are required to test the immunogenicity and the efficacy of this novel vaccine before applications using chickens and turkey. Full article

Hilsa muscle biomass was hydrolyzed to recover bioactive proteins and peptides using subcritical water hydrolysis (SWH) with ultrasound pretreatment at various temperatures (150–250 °C) and pressures (40–80 bar). Hydrolysis yield was increased with rising temperature and pressure, with the highest yield (98.70 ± 0.60%) observed at 250 °C and 80 bar using SWH with ultrasound pretreatment (UPT). The properties of the hydrolysates—including pH, taurine content, and color values—were influenced by the hydrolysis conditions. Additionally, total protein content, reducing sugar levels, antioxidant activity, foaming capacity (FC), and foaming stability (FS) increased at higher temperatures and pressures. Hydrolysates prepared at 225 °C and 70 bar, as well as at 250 °C and 80 bar, contained highly bioactive peptides and molecules that exhibited inhibitory effects on hydroxyl radicals and prevented λ DNA degradation. The highest 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) activities were observed in hydrolysates obtained at 225 °C and 70 bar with ultrasound pretreatment, reaching 96.88 ± 0.26% and 76.36 ± 2.84%, respectively. Amino acid profile analysis showed that the greatest number of amino acids (eight) was detected in hydrolysates produced through SWH at 250 °C and 80 bar. Overall, SWH combined with ultrasound pretreatment markedly enhanced yield and biofunctional properties of hilsa protein hydrolysates, highlighting their potential applications in food, pharmaceutical, and cosmetic industries. Full article

Extracellular vesicles (EVs) are lipid bilayer-delimited particles that are naturally released from all types of cells into the extracellular environment. During their release to the extracellular space, EVs adsorb biomolecules, collectively designated as EV corona. In this study, we developed a novel method for immobilizing EVs and assessing their capacity to bind biomolecules, using human serum albumin (HSA) as a model molecule for corona acquisition and optical waveguide lightmode spectroscopy as the analytical technique. Ten ng/mL poly-L-lysine (15–30 kDa), prepared in 42 mM TRIS buffer at pH 7.4, was used to achieve non-specific immobilization of EVs at a concentration of 4.5 × 10¹⁰ particles/mL, and also to provide surface passivation against proteins. The experiments were performed at 25 °C with a 100 µL/min flow rate. The optimal HSA detection range was between 1 and 100 ng/mL with EC₅₀ = 2.39 ng/mL. A mathematical model was also established to determine the estimated number of HSA molecules that can be bound on the surface of EVs. In saturation experiments it was demonstrated that the magnitude of HSA bound on the surface of EVs corresponded to the theoretical estimate. After its development, the method was further tested using human plasma and a chosen protein (DJ-1) as well as non-protein corona components, specifically double-stranded DNA and a microRNA (miR494). Full article

The Hainan gibbon (Nomascus hainanus) is one of the most threatened primates, yet its mitochondrial haplotype diversity and genealogy remain poorly understood. This study investigated mitochondrial haplotype structure, genealogical history, and selective constraints on mitochondrial protein-coding genes using complete mitochondrial genomes. We generated mitogenomic data from 13 wild individuals representing five of seven extant social groups, including the recently formed F and G groups, which were established through natural dispersal and had not previously been genetically characterized. Complete mitogenomes were obtained using a combination of Illumina and Sanger sequencing, followed by population genetic analyses, phylogenetic reconstruction, molecular dating, and mitochondrial selection tests. All mitogenomes were 16,481 bp in length and exhibited very low nucleotide diversity (π = 0.000498), with four haplotypes forming two major maternal clades. Groups B, C, and E contained haplotypes from both major maternal clades, whereas F and G each showed a single sampled haplotype, a pattern consistent with maternal founder effects in these recently formed groups. The two major maternal clades had an estimated time to the most recent common ancestor (TMRCA) of ~27 ka (95% highest posterior density (HPD): 11.9–45.7 ka), placing their most recent common ancestor in the Late Pleistocene. Interspecific selection analyses were consistent with predominantly purifying selection across mitochondrial protein-coding genes, with no significant evidence of episodic positive selection or altered selection intensity. These findings complement previous nuclear genomic and partial mitochondrial DNA studies by providing a population-level view of complete-mitogenome haplotype distribution across five sampled social groups. Full article

Phytohormones, specifically from the cytokinin family, represent a promising class of sustainable, non-immunogenic alternatives to animal-derived growth factors in skin tissue engineering. This review explores the innovative integration of selected cytokinins—namely kinetin (K), benzyladenine (BA), and traumatic acid (TA)—into biocompatible biomaterials, focusing on their distinct molecular mechanisms and regenerative capacities in human cell models. At the cellular level, these compounds comprehensively modulate cell fate, proliferation, and survival. Benzyladenine and its derivatives regulate cell cycle transitions, enhance D-type cyclin expression, and selectively inhibit phosphodiesterase-4 (PDE4), thereby suppressing tumor necrosis factor-alpha synthesis to deliver crucial intrinsic anti-inflammatory benefits. Concurrently, kinetin functions as an advanced molecular shield and potent free radical scavenger, delaying the morphological features of fibroblast senescence and protecting nuclear DNA against oxidative damage by preventing 8-oxo-dG formation. Furthermore, the plant wound hormone traumatic acid exhibits a dual biological activity; it selectively serves as a significant antioxidant that accelerates extracellular matrix remodeling by driving protein and collagen biosynthesis in normal skin fibroblasts, while shifting to a pro-oxidant, cytotoxic mechanism in cancerous cells. By merging these specialized anti-inflammatory, anti-aging, and matrix-stimulating pathways into biocompatible scaffolds or hydrogels, cytokinin-functionalized biomaterials eliminate the safety and contamination risks associated with mammalian cell products. Ultimately, this approach establishes a highly cost-effective, scalable, and multi-functional platform capable of driving advanced tissue repair, enhancing wound healing dynamics, and ensuring significant biocompatibility. Full article

Background: Calcium/calmodulin-dependent protein kinase II alpha (CAMK2A) is a neuron-enriched kinase that plays essential roles in synaptic signaling and neuronal differentiation. However, its clinical significance and biological relevance in glioma remain incompletely understood. This study aimed to systematically characterize CAMK2A across human cancers and investigate its prognostic significance and association with neuronal differentiation programs in glioma. Methods: An integrated multi-omics analysis was performed using pan-cancer transcriptomic, proteomic, genomic, DNA methylation, bulk RNA-sequencing, and single-cell RNA-sequencing datasets from TCGA, CPTAC, GTEx, CGGA, and other public resources. CAMK2A expression patterns, clinical associations, prognostic value, molecular characteristics, pathway activities, and cellular localization were systematically evaluated. Multiplex immunofluorescence staining of human glioma specimens was further performed to validate the protein-level expression patterns of CaMKII and the neuronal marker MAP2. Results:CAMK2A exhibited lineage-specific dysregulation across human cancers and was progressively downregulated during glioma progression. Higher CAMK2A expression was consistently associated with prolonged overall survival across multiple independent glioma cohorts and remained an independent favorable prognostic factor after adjustment for established clinicopathological and molecular variables. Functional enrichment analyses linked CAMK2A to neuronal differentiation, synaptic organization, calcium signaling, and neurotransmission-related pathways. Gliomas with high CAMK2A expression exhibited lower scores for several malignancy-associated pathways and enrichment of neuronal transcriptional signatures; these tissue-level associations do not distinguish tumor-intrinsic programs from normal neuronal admixture. Single-cell transcriptomic analyses further revealed preferential enrichment of CAMK2A in neuronal populations. Consistently, multiplex immunofluorescence demonstrated a coordinated reduction in CaMKII and MAP2 protein expression with glioma progression. Conclusions:CAMK2A represents a neuron-associated biomarker linked to preserved neuronal differentiation programs and favorable clinical outcomes in glioma. These findings support its potential value as a prognostic biomarker and provide further insight into the relationship between neuronal differentiation programs and glioma progression. Full article

Early diagnosis of hepatocellular carcinoma (HCC) still represents a significant challenge, particularly in the context of etiological shift toward obesity and non-viral liver disease_-associated HCC. Through intersection-based analysis of independent transcriptomic datasets comprising healthy controls, individuals with obesity or NAFLD, and HCC patients with viral or non-viral etiologies, we prioritized ADAMTS3, a secreted protein, for further investigation of its functional and mechanistic relevance to HCC. ADAMTS3 was upregulated in HCC and demonstrated transcriptomic discrimination between HCC and non-HCC populations. Notably, in the validation cohort, ADAMTS3 retained discriminative capacity in clinically defined alpha-fetoprotein-negative HCC patients. Functionally, ADAMTS3 knockdown impaired cell proliferation, clonogenicity, and migration, while inducing apoptosis and cell-cycle arrest. Reintroduction of ADAMTS3 partially rescued these cellular effects. Mechanistically, ADAMTS3 promotes HCC cell growth and migration through TGF-β:TGFBR2 signaling. ADAMTS3 knockdown increased extracellular TGF-β and downstream SMAD phosphorylation. Neutralization of extracellular TGF-β alleviated growth inhibition caused by ADAMTS3 knockdown. Consistently, TGFBR2 knockdown partially restored the cell growth and migration suppressed by ADAMTS3 knockdown. We also developed a DNA origami-based SERS aptasensor targeting ADAMTS3, achieving highly sensitive detection of ADAMTS3 down to 10⁻¹² M with strong quantitative capability (R² = 0.914). Together, our findings support a role for ADAMTS3 in promoting HCC cell growth and migration and provide a basis for further investigation of its relevance to HCC biomarker development. The DNA origami-based SERS aptasensor provides a sensitive analytical approach for future studies of circulating or serum ADAMTS3. Full article

Identifying oocytes with high meiotic competence remains a major challenge for improving in vitro embryo production in South American camelids. Although Brilliant Cresyl Blue (BCB) staining has been used to distinguish alpaca oocytes with different meiotic and developmental competence, the biological and molecular feature associated with their meiotic competence remain poorly understood. This study characterized the transcriptomic profiles of immature alpaca (Vicugna pacos) oocytes classified as BCB-positive (BCB+) or BCB-negative (BCB−) using single-cell RNA sequencing. BCB+ oocytes exhibited a significantly larger ooplasm diameter, a higher proportion of condensed chromatin configurations, and higher in vitro maturation rates than BCB− oocytes. Transcriptomic analysis identified 2978 differentially expressed genes and revealed a clear separation between groups (PC1 = 64.8%). Differentially expressed genes with higher transcript abundance in BCB+ oocytes were enriched in DNA metabolism, cell cycle, oocyte meiosis, and proteasome pathways, whereas those with higher transcript abundance in BCB− oocytes were enriched in gene expression regulation, RNA metabolism, chromatin remodeling, and intracellular signaling. Protein–protein interaction (PPI) network analysis identified distinct hub genes in each group, including CDK1, CCNB2, MAD2L1, and AURKB in BCB+ oocytes and EP300, CREBBP, SMARCA4, and ARID1A in BCB− oocytes. These results provide the first single-cell transcriptomic characterization of BCB-classified alpaca oocytes and establish the molecular basis underlying the biological differences identified by BCB staining. Full article

Folliculogenesis is essential for fish reproduction, but the role of ERK signaling during early primary follicle development in teleosts remains unclear. Here, we reanalyzed and integrated previously published transcriptomic and DNA methylation datasets from four primary follicle subtypes in zebrafish to identify MAPK-related candidate genes. No new sequencing was performed. Additional qRT-PCR and immunohistochemical analyses examined gene expression and the localization of total ERK and JNK proteins, respectively. Zebrafish were also exposed to the ERK inhibitor SCH772984 from 25 to 60 days post-fertilization to assess changes in ovarian development. The reanalysis identified MAPK-related genes with stage-associated differences in expression and DNA methylation. SCH772984 exposure was associated with reduced ovarian ERK phosphorylation and a lower proportion of advanced follicles. These results support a role for ERK signaling in the progression of early ovarian follicles in zebrafish. These observations derive from a single ERK inhibitor at nominal exposure; complementary assays could strengthen the evidence for target specificity. Full article

The cell cycle is a fundamental biological process indispensable for life, and its orderly progression is governed by a complex signaling network. Although the p38 MAPK pathway plays important roles in regulating cell proliferation across metazoans, its mechanistic role in insect cell cycle progression remains poorly characterized, particularly in species with endoreplication systems. Here, we investigated the function and mechanism of Bombyx mori p38 MAPK (Bmp38 MAPK) in cell cycle regulation using the silkworm silk gland as a specialized endoreplication model. We found that the p38 MAPK is evolutionarily conserved from insects to mammals and exhibits both cytoplasmic and nuclear localization, with its expression peaking during the G1 phase. Overexpression of Bmp38 MAPK induced cell cycle arrest at both G1/S and G2/M phases and suppressed cell proliferation, whereas Bmp38 MAPK knockdown alleviated G1/S arrest and enhanced proliferation. Mechanistically, Bmp38 MAPK increased BmCyclin B expression while reducing the expression of BmCyclin E, BmCDK2 and BmCDK1. Protein interaction assays further confirmed interactions between Bmp38 MAPK and BmCDK2, BmCyclin E and BmCyclin B. Notably, Bmp38 MAPK significantly inhibited DNA synthesis in silk gland cells, suggesting that Bmp38 MAPK contributes to the regulation of endoreplication. These findings reveal a possible role for p38 MAPK in insect cell cycle control and provide potential mechanistic insights into the regulation of endoreplication in the silkworm silk gland. Full article

Glioblastoma (GBM) is the most common and highly malignant primary brain tumor in adults. Even with advances in surgery, radiotherapy, and systemic therapies, patient outcomes remain poor due to extensive intratumoral heterogeneity and the development of therapeutic resistance. Circular RNAs (circRNAs) are a class of covalently closed RNA molecules that are highly abundant in the central nervous system and frequently dysregulated in cancer. While originally considered non-coding transcripts, current evidence demonstrates that a subset of circRNAs contain functional open reading frames and can undergo cap-independent translation to generate biologically active proteins and peptides. Recent studies have identified a growing number of translated circRNAs in GBM encoding proteins implicated in diverse processes such as receptor tyrosine kinase signaling, DNA repair, metabolism, stemness, invasion, and therapeutic resistance. These findings expand the coding potential of the GBM transcriptome and highlight an additional layer of post-transcriptional regulation. In this review, we summarize our current understanding of circRNA translation, discuss the biological functions of translated circRNAs in GBM, and examine their potential roles in tumor progression, therapeutic resistance, and clinical management. Additionally, we consider the opportunities and challenges associated with targeting translated circRNAs and their encoded proteins for the treatment of GBM. Full article

The RNA World theory was proposed as a way to explain the origin of the DNA-RNA-Protein relationship found all current cells. It has been greatly strengthened by experimental work on ribozymes and non-enzymatic replication, and has become the most widely accepted theory for the origin of life. But how much do we really know? Within the science community there remain arguments that something else is required before RNA, or that we should not focus on RNA as a single central player. From outside, there are claims that scientists are clueless about the origin of life and that biological information can only be explained by intelligent design. Here, I review recent work showing the strength and coherence of the case for the RNA World. Science can clearly defeat the argument for intelligent design. There is substantial progress with experiments demonstrating many of the requirements for an RNA-first Origin of Life, but important issues remain to be understood. Full article

The incidence of cutaneous squamous cell carcinoma (cSCC) has been rising steadily in recent years, yet treatment options for advanced-stage patients remain limited. Bee venom (BV) possesses diverse biological activities including inhibiting tumor cell growth in various tumor models, but its underlying mechanism against cSCC A431 cells remains unclear. This study evaluated the antiproliferative effect of BV on A431 cells using the CCK-8 assay and explored its mechanism of action through label-free proteomics and untargeted metabolomics. BV inhibited A431 cell proliferation in a dose-dependent manner, with a half-maximal inhibitory concentration of 11.57 μg/mL. Proteomics analysis revealed that 118 differentially expressed proteins were mainly enriched in the p53 signaling pathway, mineral absorption, and base excision repair pathways. Metabolomics screening yielded 336 differentially expressed metabolites, primarily enriched in amino acid biosynthesis and metabolism. BV exerts its anti-A431 cell bioactivity potentially by synergistically regulating tumor-related signaling pathways, mineral homeostasis, and proteins involved in DNA repair processes, thereby disrupting amino acid biosynthesis and metabolism. But the transcriptomic data failed to confirm TP53 upregulation. These findings provide a theoretical basis for apipuncture and BV-based cSCC therapies. Full article