Search Results (113)

As a naturally self-assembling protein nanocarrier, ferritin enables multivalent antigen display and functions as an intrinsic adjuvant to enhance vaccine-induced immune responses. Streptococcus equi subsp. equi (S. equi) is the causative agent of equine strangles, an acute and highly contagious respiratory disease responsible for substantial economic losses worldwide. However, currently available vaccines often show suboptimal immunogenicity and limited protective efficacy. In this study, we developed a recombinant equine ferritin (rHF)-based nanoparticle vaccine, rSE5Mix, presenting five core protective antigens (EQ8, EQ5, CNE, IdeE, EAG). The fusion proteins efficiently assembled into uniform nanoparticles. Immunization of BALB/c mice elicited rapid, high-titer antigen-specific IgG antibodies and a balanced Th1/Th2 immune response without additional adjuvants. Following lethal challenge with S. equi, rSE5Mix-immunized mice showed 100% survival and markedly milder clinical signs. Histopathological analysis demonstrated significantly alleviated organ damage, and bacterial loads in major tissues were reduced to nearly undetectable levels. Importantly, compared with the octavalent tandem vaccine rSE8, rSE5Mix induced faster elevation of partial antigen-specific antibodies, especially for EQ8 and CNE. Their antibody titers were comparable at later stages. This study developed a safe and effective ferritin nanoparticle vaccine candidate against equine strangles and verified that equine ferritin is a promising candidate delivery platform for veterinary bacterial vaccines. Full article

Ketosis is one of the most economically significant metabolic disorders affecting periparturient dairy cows, causing production losses and predisposing animals to secondary complications. Current blood-based diagnostics are invasive and provide limited insight into the underlying metabolic perturbations. This study employed an integrated three-platform metabolomics approach to characterize milk metabolite alterations in ketotic Holstein dairy cows and to evaluate milk-based biomarker panels for early ketosis detection. Milk samples from 20 healthy control (CON) cows and 6 ketotic cows were collected at 2 weeks postpartum and analyzed by direct injection/liquid chromatography–tandem mass spectrometry (DI/LC-MS/MS), proton nuclear magnetic resonance (¹H-NMR) spectroscopy, and inductively coupled plasma mass spectrometry (ICP-MS). Ketosis was confirmed by serum β-hydroxybutyrate concentrations ≥ 1400 μmol/L. Principal component analysis, partial least squares-discriminant analysis, and receiver operating characteristic (ROC) curve analyses were applied. All three platforms discriminated ketotic cows from healthy cows, with clear cluster separation validated by 2000 permutation tests (p < 0.05). DI/LC-MS/MS identified 16 significantly altered metabolites (p < 0.05), with butyrylcarnitine (C4), phosphatidylcholine 30:0 (PC 30:0), ether-linked phosphatidylcholine O-38:3 (PC O-38:3), and citrulline identified as the top discriminatory biomarkers (AUC = 0.920; 95% CI: 0.85–0.98; sensitivity = 91.7%; specificity = 93.3%). ICP-MS revealed significantly reduced selenium (Se, p = 0.017), manganese (Mn, p = 0.045), and chromium (Cr, p = 0.037), as well as elevated cobalt (Co, p = 0.014) in ketotic milk (AUC = 0.870). ¹H-NMR detected no individually significant metabolites; however, multivariate analysis distinguished groups (AUC = 0.890), with succinate (numerical fold change: +5.77×; p = 0.059), methanol (−1.94×; not significant), and acetate (+2.88×; not significant) as top VIP contributors. The combined multi-platform biomarker panel (joint classification using top VIP features from all three platforms, without formal data fusion) achieved superior diagnostic performance (AUC = 0.970; 95% CI: 0.93–1.00; sensitivity = 95.0%; specificity = 96.7%). These findings identify coordinated perturbations in glycerophospholipid metabolism, acylcarnitine profiles, amino acid homeostasis, antioxidant mineral status, and energy metabolism during early ketosis, and suggest that milk metabolomics is a promising non-invasive approach for precision dairy health monitoring, pending validation in independent cohorts. We acknowledge the small ketotic group size (n = 6) as a limitation; therefore, these findings should be considered discovery cohort observations requiring prospective validation before clinical translation. Full article

Background: Fibrolamellar carcinoma (FLC) is a rare liver malignancy affecting adolescents. FLCs harbor a DNAJB1–PRKACA gene fusion that combines heat shock protein DNAJB1 with the catalytic subunit of protein kinase A. Surgery with systemic therapy provides 5-year survivals of 30–50%, but advanced disease remains largely incurable. Three-dimensional explants from 41 FLC patients were interrogated for drug sensitivity, resistance, and synergy against cytotoxics, targeted agents, and signal transduction inhibitors. Methods: Sterile specimens from histologically confirmed FLC patients were analyzed by Ex Vivo Analysis of Programmed Cell Death (EVA/PCD™) in a CLIA-licensed laboratory. Following mechanical and enzymatic disaggregation, explants underwent 72 h drug exposure. LC₅₀ values were derived from five-point dose–response curves and compared with a database of over 10,000 human tumor analyses. Synergy was assessed by combination index. In parallel, targeted metabolomic profiling was performed in five FLC patients using tandem MS/MS. Results: Forty-one samples were analyzed. Of 24 drugs selected, tumor-cell yields were adequate for testing in 18 (75%). Single-agent activity favored vorinostat, followed by phenformin and 6-diazo-5-oxo-L-norleucine. Combinations favored gemcitabine plus oxaliplatin (GEMOX) and 5-FU plus interferon. Metabolomic analysis identified distinct signature consistent with mitochondrial dysfunction and altered polyamine metabolism. Conclusions: The present findings are exploratory, and hypothesis-generating and should not be interpreted as evidence of clinical efficacy. Prospective clinical validation and mechanistic studies will be required to further define the therapeutic relevance of these observations in fibrolamellar carcinoma. Full article

Background/Objectives: The Black Caribbean population of Honduras, also referred to locally as Negro Inglés, constitutes one of the country’s nine recognized indigenous and Afro-descendant peoples. Predominantly settled in the Bay Islands and sections of the Caribbean coast, this community traces its ancestry predominantly to West Africa and has remained culturally and linguistically distinct for more than three centuries. Despite its demographic and historical relevance, no population-specific short tandem repeat (STR) database has been established for this group. Methods: Allele frequencies for 23 autosomal STR loci were characterized in 100 unrelated Black Caribbean individuals from the department of Islas de la Bahía. DNA was extracted from blood on FTA cards and amplified with the PowerPlex Fusion 6C System (Promega Corporation). Statistical parameters were computed using Genepop v4.2, Arlequin v3.5 and GDA v1.0. Results: A total of 241 distinct alleles were detected across all 23 loci (mean 10.48 ± 3.85 alleles/locus). Expected heterozygosity ranged from 0.6541 (D13S317) to 0.9350 (SE33), with a mean of 0.8150 ± 0.0664—values consistent with a population of predominantly West African origin. No locus exhibited a significant departure from Hardy–Weinberg equilibrium after Bonferroni correction (α = 0.0022). The combined power of discrimination exceeded 99.9999% and the combined chance of exclusion surpassed 99.9999%. Conclusions: This first genetic characterization of the Honduran Black Caribbean population delivers an essential, population-specific reference dataset for forensic casework, paternity testing, and population genetics research. The data also deepen the understanding of Afro-descendant genetic diversity in Central America and constitute a critical step towards equitable forensic genetic services for all Honduran ethnic communities. Full article

Background: The Pech ethnic group, comprising approximately 6024 individuals in northeastern Honduras, represents one of the country’s smallest indigenous communities with a rich cultural heritage extending to pre-Columbian times. Despite their historical significance, no population genetic studies have been conducted on this group, and population-specific databases are essential for accurate forensic applications. Methods: Allele frequencies for 23 autosomal short tandem repeat (STR) loci were determined in 100 unrelated Pech individuals (58 females, 42 males) from communities in the departments of Olancho, Colón, and Gracias a Dios. DNA was extracted from blood samples collected on FTA cards and amplified using the PowerPlex Fusion 6C System. Statistical parameters were calculated using Genepop v4.2 and Arlequin v5.3.2.2. Results: All loci exhibited substantial polymorphism. No statistically significant deviations from Hardy–Weinberg equilibrium were detected after Bonferroni correction (α = 0.0022). Expected heterozygosity ranged from 0.4033 (TH01) to 0.8563 (FGA). The combined power of discrimination exceeded 99.9999%, and the combined chance of exclusion was 99.9999%. Conclusions: This study presents the first genetic characterization of the Pech population, providing essential reference data for forensic identification, paternity testing, and population genetics research. The dataset fills a critical gap in the Honduran forensic genetic infrastructure and contributes to understanding indigenous Central American genetic diversity, enabling accurate forensic analyses for individuals of Pech ancestry in compliance with CODIS and ESS standards. Full article

Background: The Garifunas are a distinctive Afro-indigenous community of Honduras, originating from the historical admixture of Island Carib, Arawak, and West African peoples in the seventeenth-century Caribbean. With an estimated 43,111 individuals residing primarily along the northern Atlantic coast. Their dual ancestral composition yields a genetic profile that differs meaningfully from those of other Honduran reference populations, consistent with pairwise FST comparisons with previously published Lenca and Tawahka datasets generated on the identical platform; yet no population-specific short tandem repeat (STR) reference dataset had previously been established. Methods: We genotyped 23 autosomal STR loci using the PowerPlex Fusion 6C System (Promega Corporation) in 100 unrelated Garifuna individuals (70 females, 30 males) sampled from three coastal settlements in the department of Atlántida: Triunfo de la Cruz, Ensenada, and Corozal. DNA was extracted from blood collected on FTA cards, and statistical parameters were computed using Genepop v4.2 and Arlequin v5.3.2.2. Results: A total of 217 distinct alleles were identified, with 5 to 19 alleles per locus (mean 9.43 ± 3.54). Expected heterozygosity (He) ranged from 0.6392 (D13S317) to 0.9010 (SE33), with a population mean of 0.7893. No locus deviated from Hardy–Weinberg equilibrium after Bonferroni correction (α = 0.0022). The combined random match probability was approximately 1.9 × 10⁻²⁶, and the combined chance of exclusion reached 99.99999993%. Conclusions: This study provides the first Honduran Garifuna population-specific autosomal STR reference database for precise forensic likelihood ratio estimates, kinship assessments, and population genetic studies. The Garifuna’s high diversity—consistent with their West African and Amerindian ancestry—indicates the risk of systematic bias when non-specific databases are used. Full article

The 316L stainless steel (316L SS) and high-entropy alloys (HEAs) are leading candidates for radiation-tolerant structural materials in nuclear environments. Additive manufacturing (AM) enables tailored microstructures through unique thermal histories, producing high dislocation densities and sub-grain features that act as effective sinks for irradiation-induced defects. In this work, a direct quantitative comparison of helium (He) irradiation response, particularly bubble formation, is conducted between 316L SS fabricated using laser powder bed fusion (LPBF) and CoCrFeNi HEAs fabricated by laser-directed energy deposition (LDED), both possessing a face-centered cubic (FCC) crystal structure and comparable principal elemental constituents. The samples were subjected to ex situ He ion irradiation using 200 keV He⁺ ions to a peak damage dose of 10 dpa at 25 °C, 400 °C, and 600 °C at the CINT User Facility at Los Alamos National Laboratory. Post-irradiation microstructural characterization was performed using transmission electron microscopy at the IVEM-Tandem Facility at Argonne National Laboratory. For LPBF 316L SS, the areal bubble density decreases from approximately 5.1 × 10⁴ µm⁻² at 25 °C to 2.1 × 10³ µm⁻² at 600 °C, while the mean bubble diameter increases from 2.9 nm to 37.4 nm. The CoCrFeNi HEA exhibits a similar trend but retains a higher areal bubble density at elevated temperatures, with values of 2.1 × 10⁴ µm⁻² at 400 °C and 3.7 × 10³ µm⁻² at 600 °C, along with a larger mean bubble size at 400 °C compared to 316L SS. These results highlight the combined roles of AM-induced microstructures, alloy compositions, and irradiation temperatures in governing He damage evolution in FCC alloys, providing guidance for the development of radiation-tolerant materials for advanced nuclear energy applications. Full article

Endothelial cells are key regulators of vascular homeostasis, and their dysfunction plays a central role in the development of atherosclerosis and other cardiovascular diseases. Multinucleated variant endothelial cells (MVECs) have been described in pathological vascular regions; however, their functional properties remain poorly characterized. The aim of the present study was to compare lipid handling, inflammatory activation, barrier-associated features, and secretory profiles of typical endothelial cells (TECs, EA.hy926 line) and MVECs under low-density lipoprotein (LDL) exposure. MVECs were generated by polyethylene glycol-induced fusion of EA.hy926 cells and incubated with LDL under standardized conditions. Intracellular cholesterol accumulation was assessed biochemically, cytokine secretion was quantified by ELISA, gene expression of inflammatory, endothelial, junctional, and vasoactive markers was analyzed by quantitative real-time PCR, and the endothelial secretome was characterized using data-independent acquisition liquid chromatography–tandem mass spectrometry (DIA-LC-MS). MVECs demonstrated enhanced cholesterol accumulation compared with TECs following LDL exposure. At the transcriptional level, MVECs were characterized by elevated basal expression of proinflammatory markers, including IL1B, IL6, and NFKB1, and showed a markedly amplified IL6 and IL8 response to LDL. In parallel, MVECs exhibited reduced expression of genes associated with antioxidant defense (SOD1), barrier integrity (TJP1), and hemostatic function (VWF). Consistent with transcriptional data, mass spectrometry-based secretome analysis revealed decreased secretion of von Willebrand factor (vWF), vascular endothelial growth factor C (VEGFC), and endothelin-1 (EDN1) by MVECs, accompanied by increased secretion of tissue-type plasminogen activator (t-PA). Functional enrichment analysis of secretome-associated proteins highlighted pathways related to extracellular matrix–receptor interaction, focal adhesion, cell adhesion molecules, complement and coagulation cascades, and leukocyte transendothelial migration. In contrast, TECs demonstrated a more pronounced transcriptional response in EDN1, consistent with their role in vascular tone regulation. Immunocytochemical analysis further revealed altered subcellular distribution of the tight junction protein ZO-1 in MVECs, indicating junctional destabilization. Taken together, these results indicate that MVECs represent a distinct endothelial phenotype characterized by enhanced lipid accumulation, sustained proinflammatory activation, altered secretory signaling, and reduced barrier and hemostatic potential. Such features suggest that MVECs may contribute to the maintenance of chronic endothelial dysfunction and vascular inflammation under conditions of lipid overload. Full article

Background: DNA methylation profiling has become an important diagnostic tool in pediatric neuro-oncology, particularly for tumors with overlapping morphology or unusual immunophenotypes. Methods: We report four pediatric brain tumor cases evaluated by histopathology and immunohistochemistry, supplemented by targeted next-generation sequencing (NGS) and DNA methylation profiling using the DKFZ Brain Tumor Classifier (v12.8); one case also underwent DKFZ Sarcoma Classifier analysis (v13.2). Results: Methylation profiling provided clinically meaningful diagnostic insights across all cases. In two patients, methylation results supported integrated interpretation in diagnostically challenging settings without changing management. In two cases, methylation profiling prompted major diagnostic revisions with significant therapeutic consequences: (i) a tumor initially diagnosed as atypical teratoid/rhabdoid tumor was reclassified as CNS Ewing sarcoma, confirmed by an EWSR1-FLI1 fusion and immunophenotype, leading to a change to Ewing-based therapy; and (ii) a tumor interpreted as high-grade astrocytoma/glioblastoma was reclassified as a CNS tumor with BCOR internal tandem duplication, enabling a curative-intent approach and revised prognostic counseling. Conclusions: These cases illustrate that DNA methylation profiling can complement histopathology, resolve diagnostically ambiguous tumors, and in selected patients substantially alter treatment decisions and expected outcomes. Early integration of methylome profiling may improve precision diagnostics and reduce the risk of inappropriate therapy in pediatric brain tumors. Full article

Background and Clinical Significance: Non-bacterial thrombotic endocarditis (NBTE), historically termed marantic endocarditis, is a severe manifestation of cancer-associated hypercoagulability characterized by sterile valvular vegetations and a high risk of systemic embolization. While direct oral anticoagulants (DOACs) have become the standard of care for cancer-associated venous thromboembolism (CAT), their efficacy in preventing high-shear arterial thrombosis in NBTE has been contested. Emerging data suggest that DOACs may fail to halt vegetation growth in active malignancy, necessitating a reversion to heparin-based therapies. Case Presentation: A 47-year-old female with metastatic RET fusion-positive non-small cell lung cancer (NSCLC) presented with progressive dyspnea and digital ischemia despite strict adherence to therapeutic anticoagulation with rivaroxaban for a prior pulmonary embolism. Echocardiography showed large vegetations on all three cusps of the aortic valve, confirming NBTE. Computed tomography revealed extensive tumor progression. The therapeutic strategy involved an immediate switch from rivaroxaban to therapeutic low-molecular-weight heparin (LMWH) and the initiation of dual targeted therapy with selpercatinib and tepotinib. Serial transesophageal echocardiography documented regression within two weeks and eventual complete resolution of the valvular vegetations after eight weeks, occurring in tandem with a rapid radiological response of the tumor. Conclusions: Upon diagnosis of NBTE, a rapid oncologic work-up is warranted, as ongoing tumor progression is highly likely. This case questions the appropriateness of direct oral anticoagulants in patients with NBTE and active, progressive malignancy. Full article

APETALA2/ethylene-responsive factor (AP2/ERF) transcription factors integrate phytohormone signalling with developmental programmes and specialised metabolism, yet their family-wide features and potential contributions to phenolic-acid biosynthesis remain to be systematically clarified in Salvia miltiorrhiza. In this study, we conducted a comprehensive genome-wide analysis and identified 169 SmAP2/ERF genes, which were classified into five subfamilies (AP2, ERF, DREB, RAV and Soloist). SmAP2/ERFs were unevenly distributed across chromosomes and expanded predominantly through tandem and segmental duplication, and Ka/Ks analysis indicated that tandem-duplicated pairs have mainly undergone purifying selection. Promoter analysis revealed abundant cis-acting elements related to light, phytohormones and stress responses, indicating extensive regulatory potential. Comparative phylogenetic analysis with Arabidopsis thaliana prioritised four candidates (SmAP2/ERF88, SmAP2/ERF110, SmAP2/ERF121 and SmAP2/ERF122) closely associated with specialised-metabolism regulators. These genes exhibited distinct tissue-preferential expression patterns and divergent hormone responsiveness: SmAP2/ERF88/110 were broadly inducible, whereas SmAP2/ERF121/122 responded mainly to abscisic acid and were repressed by brassinosteroids. Confocal imaging of GFP fusions confirmed nuclear localisation of all four proteins. Phytohormone treatments differentially regulated key phenolic-acid pathway genes (PAL, C4H, 4CL, TAT, HPPR, RAS and CYP98A14) and altered rosmarinic acid and salvianolic acid B accumulation. These results broaden the genome-wide understanding of the SmAP2/ERF family in Salvia miltiorrhiza. Hormone-responsive SmAP2/ERFs show expression patterns associated with hormone-dependent transcriptional changes in phenolic-acid pathway genes and with RA and SAB accumulation, providing candidates for future functional validation and metabolic engineering. Full article

This study examines how the seven pillars of ZTA are implemented in a CDS environment that demands high security reliability, similar to the defense and finance sectors, and identifies the technological advancements and integration patterns that emerge during this process. With the introduction of user- and device-centric authentication methods like distributed identity and RF fingerprinting in the Identity and Device areas, there is a growing trend towards strengthening trust even in domains where distrust is prevalent. In the Network and Application domains, the focus is on using micro-segmentation and SDN to segment and control internal traffic flows, while dynamically enforcing the principle of least privilege. In the Data, Visibility, and Orchestration domains, AI analysis is being applied in real-time, leveraging log and visibility data, and orchestration is automating policy execution and response. In conclusion, it is clear that each pillar of ZTA operates in tandem with the others, rather than as isolated components within the CDS environment. This fusion structure demonstrates its ability to function as a unified security strategy that balances trust with comprehensive coverage of diverse domains. Full article

Current mainstream remote sensing approaches to forest age estimation frequently neglect interspecific differences in functional traits, which may limit the accurate representation of species-specific tree growth strategies. This study develops and validates a technical framework that incorporates multi-source remote sensing and tree species functional trait heterogeneity to systematically improve the accuracy of plantation age mapping. We constructed a processing chain—“multi-source feature fusion–species identification–heterogeneity modeling”—for a typical karst plantation landscape in southeastern Yunnan. Using the Google Earth Engine (GEE) platform, we integrated Sentinel-1/2 and Landsat time-series data, implemented a Gradient Boosting Decision Tree (GBDT) algorithm for species classification, and built age estimation models that incorporate species identity as a proxy for the growth strategy heterogeneity delineated by the Plant Economic Spectrum (PES) theory. Key results indicate: (1) Species classification reached an overall accuracy of 89.34% under spatial block cross-validation, establishing a reliable basis for subsequent modeling. (2) The operational model incorporating species information achieved an R² (coefficient of determination) of 0.84 (RMSE (Root Mean Square Error) = 6.52 years) on the test set, demonstrating a substantial improvement over the baseline model that ignored species heterogeneity (R² = 0.62). This demonstrates that species identity serves as an effective proxy for capturing the growth strategy heterogeneity described by the Plant Economic Spectrum (PES) theory, which is both distinguishable and valuable for modeling within the remote sensing feature space. (3) Error propagation analysis demonstrated strong robustness to classification uncertainties (γ = 0.23). (4) Plantation structure in the region was predominantly young-aged, with forests aged 0–20 years covering over 70% of the area. Despite inherent uncertainties in ground-reference age data, the integrated framework exhibited clear relative superiority, improving R² from 0.62 to 0.84. Both error propagation analysis (γ = 0.23) and Monte Carlo simulations affirmed the robustness of the tandem workflow and the stability of the findings, providing a reliable methodology for improved-accuracy plantation carbon sink quantification. Full article

Uterine mesenchymal tumors encompass a diverse and diagnostically challenging group of neoplasms, including smooth muscle tumors, endometrial stromal tumors (ESS), perivascular epithelioid cell tumors (PEComas), inflammatory myofibroblastic tumors (IMTs), uterine tumor resembling ovarian sex cord tumor (UTROSCT), along with many other relatively rare entities. Traditionally classified by histomorphology and immunophenotype, these tumors are now increasingly defined by recurrent genetic alterations that refine diagnosis and elucidate tumorigenesis. For example, leiomyosarcomas display complex genomic instability with frequent TP53, RB1, and ATRX mutations. Low grade-ESS are characterized by JAZF1::SUZ12 and other related fusions, whereas high-grade tumors harbor YWHAE::NUTM2 or ZC3H7B::BCOR fusions, and BCOR internal tandem duplication (ITD) alterations. PEComas frequently contain TSC1 or TSC2 mutations, leading to aberrant activation of the mTOR pathway. Beyond their diagnostic utility, these molecular signatures increasingly inform prognosis and highlight potential therapeutic targets, including CDK4/6 inhibition, PI3K/AKT/mTOR blockade, and immunotherapy. This review summarizes the evolving molecular landscape of uterine mesenchymal tumors, underscoring the value of integrating molecular testing into clinical practice to enhance diagnostic precision and enable personalized management of these rare yet clinically significant neoplasms. Full article

Background: The Tawahka ethnic group, with approximately 2690 individuals in northeastern Honduras, represents one of the country’s smallest indigenous communities. No genetic studies have been published on this population, and population-specific databases are essential for forensic applications. Methods: Allele frequencies for 23 autosomal short tandem repeats (STRs) loci were analyzed in 100 unrelated Tawahka individuals (61 females, 39 males) from the municipality of Wampusirpi. Deoxyribonucleic acid (DNA) was extracted from blood on Fast Technology for Analysis of nucleic acids (FTA) cards and amplified using the PowerPlex Fusion 6C System. Statistical parameters were calculated using Genepop v4.2 and Arlequin v5.3.2.2. Results: All loci showed substantial polymorphism with no Hardy–Weinberg equilibrium deviations after Bonferroni correction (α = 0.0022). Expected heterozygosity ranged from 0.4968 to 0.8812. Combined power of discrimination was 99.9999% and combined chance of exclusion was 99.99%. Conclusions: This first genetic characterization of the Tawahka population provides essential reference data for forensic identification, paternity testing, and population genetics studies. The dataset contributes to understanding indigenous Central American genetic diversity and ensures accurate forensic analyses for individuals of Tawahka ancestry following Combined DNA Index System (CODIS) and European Standard Set (ESS) standards. Full article