Search Results (815)

Flower color is a key ornamental trait in Nobile-type Dendrobium, yet the molecular basis underlying purple flower formation remains poorly understood. In this study, a white-flowered paternal cultivar and its purple-flowered filial line were used to investigate the regulatory mechanism of purple floral pigmentation. Comparative phenotypic analysis showed that floral color divergence was established early during flower development, and anthocyanidin profiling of full-bloom petals revealed that purple flowers accumulated substantially higher levels of most anthocyanidins than white flowers, with delphinidin- and cyanidin-derived anthocyanidins together accounting for 80.26% and 94.56% of the total anthocyanidins in white- and purple-flowered materials, respectively. Transcriptome profiling identified a total of 21,235 differentially expressed genes (DEGs), with significant enrichment of phenylpropanoid- and flavonoid-related pathways, and MYB transcription factors prominently represented among the candidate regulators across the three comparison groups. Among them, DntMYB1 was identified as a C1 subgroup R2R3-MYB associated with floral pigmentation, and transient overexpression assays in Phalaenopsis hybrid V3 and a Nobile-type Dendrobium hybrid supported its positive role in visible purple pigmentation. By integrating RNA-seq and DAP-seq analyses, we identified 3205 candidate downstream targets of DntMYB1 and established a DntMYB1-centered regulatory module. Among these candidates, one Dnt4CL1 gene and one DntF3′H1 gene were validated as robust direct targets of DntMYB1 through yeast one-hybrid, EMSA, and dual-luciferase assays. These findings suggest that DntMYB1 is associated with purple flower formation by coordinately regulating both upstream precursor metabolism and downstream anthocyanin biosynthesis, providing new insight into the molecular regulation of flower color in Nobile-type Dendrobium and useful candidate genes for ornamental trait improvement. Full article

Conifer-derived essential oils have gained attention as versatile natural additives with potential applications in animal production, including influencing microbial processes and supporting environmental sustainability. This study aimed to characterize the chemical composition of selected conifer essential oils (EOs), evaluate their antimicrobial activity against rumen microorganisms in vitro, and assess the effects of Pinus sylvestris essential oil on rumen fermentation and methane production under in vitro and in vivo conditions. EOs from Thuja occidentalis, Cupressus sempervirens, Juniperus communis, Picea mariana, Pinus sylvestris, and Pinus pinaster were analyzed by GC–MS, and their inhibitory activity against selected rumen bacteria was determined by MIC and IC₅₀ assays. Based on these results, P. sylvestris oil was selected for fermentation experiments. Ninety-two volatile compounds were identified, with monoterpenes as the dominant constituents and α-pinene as the major compound in P. sylvestris oil. In vitro, P. sylvestris oil influenced fermentation in a dose-dependent manner without affecting ruminal pH. In vivo, ruminal pH, ammonia-related parameters, and total VFA concentration were not significantly affected by treatment, whereas several variables showed a significant effect of time. Temporal changes in VFA profiles suggested a transient adaptation of ruminal fermentation. Methane concentration was significantly (p < 0.01) reduced by Pinus sylvestris essential oil supplementation, with a decrease of approximately 28.7% after 14 days. These findings indicate that P. sylvestris EOs may serve as a promising natural modulator of rumen fermentation, although further studies are needed to optimize dosage and confirm long-term effects. Full article

Pigs play a key role in the ecology of influenza A viruses (IAVs), particularly in avian influenza (AI)-endemic regions where co-circulation of viruses from different hosts increases reassortment risk. Between September 2023 and August 2024, we surveyed pigs from Lebanon and Egypt to study IAV ecology in AI-endemic countries. Nasal swabs and sera were collected and tested using real-time RT-PCR and hemagglutination inhibition assays against avian, swine, and human seasonal IAVs. Molecular analyses identified IAV-infections in both countries, including human H1 and avian H5 subtypes, which may reflect potential cross-species transmission from humans and birds. Serologic analyses revealed prior exposure to avian, swine, and human IAVs. Avian virus seropositivity reached 4.6% (H5N1) and 15.2% (H9N2) in Egypt and 8.6% (H5N1) and 4.3% (H9N2) in Lebanon. Antibodies against human H1N1 and H3N2 were prevalent in both countries. Serologic evidence exceeded molecular detection, indicating frequent past or transient infections not captured by PCR alone. Antibody responses were significantly associated with host-level factors such as housing type, age, shaping exposure risk. These findings demonstrate repeated multisource exposure of pigs to genetically distinct IAVs in AI–endemic countries, supporting the need for integrated virologic and serologic surveillance within a One Health framework. Full article

Systematic surveillance of World Organization for Animal Health (WOAH)-listed protozoan parasites is essential for maintaining the sanitary status of seafood exports and detecting the introduction of exotic pathogens into coastal ecosystems. In 2023, we examined wild Mediterranean mussels Mytilus galloprovincialis and Pacific oysters Crassostrea gigas collected from small harbors adjacent to ten major trading ports along the west and south coasts of Korea to assess the occurrence of WOAH-listed protozoan parasites and emerging Perkinsus species. A total of 1080 mussels and 1080 oysters from 18 sites were sampled in spring and autumn, and gill and digestive gland tissues were pooled from six individuals for DNA extraction. Species-specific PCR assays targeting Perkinsus marinus, P. olseni, P. beihaiensis, Bonamia ostreae, B. exitiosa, and Marteilia refringens were performed using previously validated primer sets and positive controls. All PCR assays were negative for the six protozoan parasite species in both host species across all sampling sites, indicating no detectable infections in port-adjacent wild mussel and oyster populations during the survey period. These negative results contrast with recent reports of P. marinus in wild C. gigas and B. ostreae in Ostrea denselamellosa on the west coast of Korea, suggesting that infections may currently be focal, transient, and host-specific rather than widespread in port-associated M. galloprovincialis and C. gigas populations. The present study provides baseline data on the distribution of protozoan parasites in bivalves inhabiting high-risk harbor environments and underscores the need for continued surveillance of transboundary shellfish diseases that is closely coupled with environmental monitoring. Full article

Ferroptosis is an iron-dependent, lipid peroxidation–driven form of regulated cell death that has emerged as a therapeutic vulnerability in hepatocellular carcinoma (HCC), yet the contribution of lysosomes to this process remains incompletely understood. In this study, we investigated whether lysosomal ion channels regulate ferroptosis sensitivity in HCC cells, focusing on the two-pore channel 2 (TPC2) and the transient receptor potential mucolipin 1 (TRPML1). Using pharmacological modulation, genetic knockout models, flow cytometry-based cell death and lipid peroxidation assays, lipidomics, calcium measurements, and molecular analyses across multiple HCC cell lines, we examined how these channels influence ferroptotic signaling. We show that NAADP-dependent TPC2 activity is required for efficient ferroptosis induction, whereas TPC2 loss renders HCC cells resistant to ferroptosis triggered by system Xc⁻ inhibition or glutathione peroxidase 4 (GPX4)blockade. This resistance is associated with reduced lipid peroxidation, altered calcium signaling, and selective depletion of polyunsaturated phosphatidylethanolamine species linked to decreased Acyl-CoA Synthetase Long-Chain Family Member 4 (ACSL4) expression. In contrast, TRPML1 deficiency sensitizes cells to ferroptosis and correlates with enhanced endoplasmic reticulum stress and oxidative imbalance rather than major lipid remodeling. Collectively, these findings identify lysosomal ion channels as key modulators of ferroptosis in HCC and highlight distinct mechanisms by which TPC2 and TRPML1 regulate cellular redox balance and death susceptibility. Full article

Anthocyanin biosynthesis in eggplant (Solanum melongena L.) is highly light-dependent, and insufficient light severely impairs fruit coloration, which restricts the development of the eggplant industry. SmMYB75 is a key positive regulator of anthocyanin biosynthesis, but its regulatory partners remain unclear. In this study, seven SmYABBY genes were identified from the eggplant genome, all containing conserved zinc finger and YABBY domains. Expression analysis showed that SmYABBY1 was predominantly expressed in fruit peel and significantly induced by light, with a peak at 4 h after light exposure. The yeast two-hybrid and bimolecular fluorescence complementation assays indicated that SmYABBY1 interacts with SmMYB75 and the light signaling regulator SmCOP1 in the nucleus. The heterologous overexpression of SmYABBY1 in Arabidopsis enhanced anthocyanin accumulation and upregulated the expression of anthocyanin structural genes. Transient co-expression in tobacco leaves further demonstrated that SmYABBY1 synergistically enhances SmMYB75-mediated anthocyanin biosynthesis. The yeast one-hybrid and Dual-LUC assays revealed that SmYABBY1 does not directly bind to the promoters of SmMYB75, SmDFR, and SmANS but indirectly promotes their transcriptional activity. Our results illustrate that SmYABBY1 acts as a transcriptional co-activator, interacting with SmMYB75 to promote anthocyanin accumulation, while SmCOP1 is involved in this regulatory process. This study provides a molecular basis for improving eggplant coloration under suboptimal light conditions. Full article

During production, fish are exposed to multiple environmental, physiological, and physical stressors, which compromise development, productivity, and welfare and urge the implementation of effective and safe stress-mitigating strategies, particularly during early developmental stages. Larval zebrafish (Danio rerio) constitute a powerful model for studying acute stress responses due to the numerous advantages they offer, such as developmental transparency, a conserved hypothalamic–pituitary–interrenal (HPI) axis, and suitability for high-throughput screening. This review examines the potential of natural monoterpenes as stress-reducing compounds and compares their performance with conventional synthetic anesthetics. Evidence from vortex-flow stress paradigms, behavioral profiling and biochemical assays shows that acute stress in zebrafish larvae triggers metabolic disruption, behavioral hyperactivity and enzyme imbalance, with cortisol responses depending on stimulus intensity. Monoterpenes such as thymol and menthol consistently reduce stress-induced hyperactivity, support redox homeostasis and display favorable safety profiles at low doses and short exposures. Nevertheless, as research into these substances is still recent, evidence of any potential adverse effects is still limited. Although individual monoterpenes may act on different subsets of molecular targets, their multimodal mechanisms, including gamma-aminobutyric acid (GABA)ergic enhancement, voltage-gated ion channel and transient receptor potential (TRP) modulation, suggest broader and potentially safer actions compared to single-target anesthetics as tricaine methane sulfonate (MS-222). Collectively, these findings suggest that monoterpenes offer promising natural alternatives for stress mitigation in aquaculture and the refinement of research procedures involving early life stages. Full article

Enterococcus lactis is increasingly recognized as an emerging mastitis pathogen, yet the functional basis of its virulence and associated health risks remain poorly defined. This study presents an integrated genomic and phenotypic characterization of E. lactis strain EL-A150 isolated from bovine subclinical mastitis. Whole-genome sequencing revealed a 2.49 Mb circular chromosome encoding multiple genes associated with adhesion (acm, bepA, fms, sagA), biofilm formation (empB, empC) and antimicrobial resistance, including determinants related to aminoglycosides and macrolides. Phenotypic assays demonstrated rapid growth, strong biofilm-forming capacity and high adhesion to bovine mammary epithelial cells, while internalization remained low and intracellular persistence was transient. Comparative genomic analyses confirmed the taxonomic placement of the strain within the E. lactis clade (ANI up to 99.5% against reference genomes) and revealed a limited resistome composed of chromosomally encoded genes, with no detectable plasmids or major mobile genetic elements. Collectively, these findings demonstrate that E. lactis EL-A150 possesses a coordinated set of traits conducive to intramammary colonization, supporting its classification as an opportunistic pathogen. The convergence of virulence potential and clinically relevant antimicrobial resistance within a single isolate underscores a One Health concern and highlights the need for surveillance frameworks that integrate functional validation with genomic risk assessment. Full article

Seed oils from Sicilian white (WGSO) and red grapes (RGSO) were examined for their possible cytotoxic effect on HepG2 liver and CaCo-2 colorectal cancer cells, the latter also induced to intestinal differentiation. Half maximal inhibitory dilution (ID₅₀) values were obtained from viability assays, excluding RGSO-treated HepG2 and differentiated CaCo-2 cells exposed to both oils, which were unresponsive. Cell morphology and cycle status, reactive oxygen species (ROS) production, and the levels of cytoprotection, regulated cell death (RCD), and autophagy markers were evaluated. No occurrence of canonical apoptosis was proven in any experimental condition. In HepG2 cells, WGSO ID₅₀ primarily triggered autophagy collapse, as evidenced by modulation of Beclin-1, p62 and LC3 markers, initiating a cascade of metabolic disturbances that led to oxidative stress reaction and mild inflammatory signaling. In CaCo-2 cells, WGSO ID₅₀ mainly elicited a strong ROS-mediated cell injury without major alterations in autophagy, with transient activation but incomplete execution of pyroptotic and necroptotic effectors (gasdermin-D, pMLKL and HMGB1). In the same cells, RGSO ID₅₀ induced a weaker metabolic perturbation with transient activation of multiple RCD pathways and concomitant autophagy inhibition. Research findings revealed distinct damage-inducing properties linked to oils’ chemical profiles, underscoring their prospective utilization as beneficial bioactive supplements. Full article

Low-salinity stress poses a critical constraint on commercial aquaculture and the survival of the sea cucumber (Apostichopus japonicus). This study investigated the regulatory network involving lncRNA011760, miR-novel-91, and their target gene NIPA2 in response to salinity fluctuations. Using integrated in vivo and in vitro functional assays, we demonstrate that lncRNA011760 acts as a competitive endogenous RNA (ceRNA) to sponge miR-novel-91, thereby alleviating the post-transcriptional repression of NIPA2. Based on these molecular dynamics, we propose a novel inhibition-adaptation-survival three-stage model. Initially (0–3 h), acute NIPA2 upregulation enhances Mg²⁺ transport efficiency to mitigate osmotic shock. During the mid-stage (3–24 h), miR-novel-91-mediated NIPA2 suppression creates a transient biosynthetic window, facilitating a shift from passive tolerance to active metabolic adaptation. Ultimately (24–48 h), lncRNA-driven NIPA2 restoration sustains Mg²⁺ homeostasis, allowing the organism to enter a low-metabolism survival mode. These stage-specific shifts reflect the inherent physiological strategies of sea cucumbers as osmoconformers. Full article

Rapid alkalinization factor (RALF) peptides are recognized as multifunctional regulators of plant stress responses, yet their roles in woody species remain poorly defined. Here, we identified a RALF22-like peptide from poplar ‘Shanxin’ (Populus davidiana × P. bolleana; PdbRALF22-like) and investigated its roles in salt tolerance and disease resistance. Synthetic PdbRALF22-like peptide elicited a rapid ROS burst in poplar leaf discs. In Nicotiana benthamiana, which was otherwise unresponsive to the peptide, transient expression of either of two poplar FERONIA-like receptor kinases (PdbFER-like-1 and PdbFER-like-2) enabled peptide-triggered ROS production, consistent with receptor-matched responsiveness in a heterologous context. Using CRISPR/Cas9, we generated a PdbRALF22-like knockout line and assessed salt tolerance in vitro and soil-grown assays. Under salinity, the mutant showed sustained rooting at high NaCl concentrations and improved growth relative to wild type. After 0.2 M NaCl treatment, soil-grown mutant plants exhibited reduced wilting and leaf injury. Evans Blue, DAB, and NBT staining indicated reduced membrane damage and lower accumulation of hydrogen peroxide and superoxide in the mutant. Significantly, the same knockout line displayed increased susceptibility to infection by the poplar leaf spot fungus, with larger lesions and higher pathogen biomass, accompanied by reduced ROS output and lower induction of the defense marker gene PdbPR1. Collectively, PdbRALF22-like negatively regulates salt tolerance while contributing positively to disease resistance, and represents a regulatory node linking salinity tolerance and disease susceptibility in poplar ‘Shanxin’, with poplar FER-like receptors providing a plausible route for peptide-triggered ROS signaling. This work expands our understanding of RALF peptide signaling in woody plants. Full article

Papain-like cysteine proteases (PLCPs) are central immune hubs frequently targeted by pathogen effectors. Sugarcane smut, caused by Sporisorium scitamineum, threatens global sugarcane yield, yet effector manipulation of host PLCPs remains unclear. Genome-wide analysis of Saccharum spontaneum AP85-441 identified 61 PLCP-encoding genes, which were classified into nine conserved subfamilies. Among these, ScRD21A, a member of the RD21 subfamily, was prioritized for functional characterization. Two Pit2 homologs, SsPit2A and SsPit2B, were identified from S. scitamineum. Yeast two-hybrid, BiFC and pull-down assays demonstrated that both effectors interact with ScRD21A, and that this interaction depends on a conserved LXRR motif within their PID14-like region. In total protein extracts from Nicotiana benthamiana, co-expression of SsPit2A or SsPit2B reduced ScRD21A-associated cysteine protease activity. Transient expression of ScRD21A enhanced flg22-induced ROS production, attenuated Pst DC3000-induced hypersensitive response-associated necrosis, and increased resistance to Botrytis cinerea. Together, these results support a conserved PLCP-targeting strategy in smut fungi and identify the ScRD21A–SsPit2A/B module as a tractable framework for studying effector–protease interactions relevant to sugarcane smut. Full article

Functional antibody-dependent enhancement (ADE) represents a fundamental and context-dependent characteristic of antiviral antibody responses, reflecting the dual capacity of antibodies to mediate both the neutralization and Fc receptor-dependent enhancement of infection. In flavivirus research, this duality complicates the interpretation of conventional serological metrics and limits the reliability of single-parameter correlates of immunity, particularly in populations with complex exposure histories. Over the past decade, functional ADE assays have evolved from specialized mechanistic tools into integrated immune assessment platforms supporting translational immunology, vaccine evaluation, and population-level immune surveillance. These platforms incorporate Fcγ receptor-relevant target cell systems, standardized viral inputs, dilution series-based profiling, quantitative enhancement metrics, and structured quality control frameworks to enable reproducible, comparable, and context-aware functional measurements across cohorts and laboratories. A central concept emerging from these developments is that ADE reflects a dynamic functional immune state rather than an intrinsic property of antibodies or a direct indicator of pathological risk. Accordingly, functional ADE platforms support the contextual interpretation of antibody activity across physiologically relevant conditions, facilitating discrimination between transient functional enhancement and clinically meaningful immunological risk. By integrating functional ADE metrics with serological, cellular, and epidemiological data, these platforms provide a structured framework for interpreting immune profiles in vaccine evaluation, booster strategy design, and population-level risk stratification. This review synthesizes the development, standardization, and global dissemination of functional ADE platforms and discusses key principles governing biological relevance, analytical robustness, and inter-site transferability. Emerging directions integrating functional ADE profiling with systems immunology, immunogenomics, and computational modeling are highlighted as pathways toward predictive, decision-support-oriented frameworks. By positioning ADE platforms as immune assessment infrastructures rather than isolated assays, this review underscores their value for mechanistic inquiry, translational interpretation, and preparedness-oriented responses to emerging viral threats in the absence of definitive correlates of protection. Full article

Meningovascular syphilis, a type of neurosyphilis, causes stroke and various types of myelopathy. In recent years, there has been an increase in the incidence of neurosyphilis. However, diagnosing neurosyphilis remains challenging due to the reliance on serum and cerebrospinal fluid (CSF) testing, which has low specificity and sensitivity. Magnetic resonance vessel wall imaging (MR-VWI), recently developed to identify vessel wall pathologies, may aid in diagnosing neurosyphilis. In this cross-sectional study, we performed systematic screening for syphilis in all 366 patients with acute ischemic stroke or transient ischemic attack admitted to our stroke unit. Further CSF analysis and MR-VWI were specifically conducted only on those with reactive serum venereal disease research laboratory (VDRL) or treponema pallidum particle hemagglutination assay (TPHA) tests to evaluate neurosyphilis. Serum screening was reactive in 5.7% (21/366) of patients; among these, the prevalence of likely neurosyphilis (defined by abnormal CSF pleocytosis or protein levels) was 2.2% (8/366). Within this group of eight patients, MR-VWI was technically feasible and thus performed in six cases. Although all CSF-VDRL tests were non-reactive, MR-VWI identified diagnostic evidence of meningovascular syphilis (concentric wall thickening and enhancement) in 33.3% (2/6) of symptomatic patients who underwent the scan. Neurosyphilis remains a critical, treatable cause of stroke that can affect older patients with established vascular risk factors. Our findings demonstrate that routine serum screening is essential, as traditional CSF-VDRL tests may yield false-negative results. MR-VWI serves as a valuable adjunct tool to provide objective evidence of active vasculitis, guiding the initiation of appropriate antibiotic therapy when laboratory results are inconclusive. Full article

Background/Objectives: Bladder cancer treatment is frequently hindered by chemoresistance to agents such as cisplatin, a process in which autophagy is hypothesized to play a cytoprotective role. This study aimed to investigate the time-dependent transcriptional dynamics of autophagy-related genes in response to cisplatin in bladder cancer cell lines to better elucidate the molecular underpinnings of this resistance. Methods: Two human bladder cancer cell lines, T24 and 5637, were exposed to varying concentrations of cisplatin. Cell viability and half-maximal inhibitory concentration (IC₅₀) values were determined at 24 and 48 h using the MTS assay. Subsequently, the relative mRNA expression levels of key autophagy-related genes (ULK1, BECN1, ATG5, ATG7, LC3B, SQSTM1/p62, LAMP1, and TFEB) were quantitatively analyzed via RT-qPCR at 0, 6, 24, and 48 h intervals. Results: Cisplatin exerted a dose- and time-dependent cytotoxic effect, with 5637 cells exhibiting significantly greater sensitivity compared to T24 cells. Transcriptional analysis revealed a dynamic, multiphasic modulation of the autophagic pathway: an early-phase upregulation of initiation genes (ULK1, BECN1), a mid-phase increase in autophagosome formation genes (ATG5, ATG7), and a late-phase alteration in lysosomal regulation genes (LAMP1, TFEB). Notably, the more chemoresistant T24 cells mounted a robust and sustained autophagic transcriptional response, whereas the sensitive 5637 cells demonstrated a more limited and transient reaction. Conclusions: Cisplatin modulates the autophagic pathway at the transcriptional level in a highly dynamic, time-dependent, and cell-line-specific manner. Interpreted alongside established functional evidence in the literature, the sustained autophagic gene expression observed in the resistant cells is consistent with a potential cytoprotective role, warranting further functional validation at the protein level. These findings map the temporal genetic landscape of cisplatin-induced autophagy, providing a theoretical framework for optimizing the timing of autophagy-targeted combination therapies in bladder cancer. Full article