Search Results (6,473)

Unc-51-like kinase 3 (ULK3) is a key member of the ULK serine/threonine kinase family. Aberrant ULK3 expression has been increasingly linked to tumorigenesis and malignant progression in multiple cancer types. However, the precise role of ULK3 in tumor initiation and progression remains incompletely understood. Leveraging integrated multi-omics data from The Cancer Genome Atlas (TCGA), the Genotype-Tissue Expression (GTEx) project, and the Clinical Proteomic Tumor Analysis Consortium (CPTAC), we systematically characterized the expression of ULK3 at both the transcript and protein levels across 33 cancer types. We also evaluated genomic alterations, prognostic significance, alternative splicing, pathway enrichment, tumor stemness, immune infiltration, and immunotherapy-related biomarkers. In parallel, we investigated the function of ULK3 in prostate cancer PC-3 cells using cellular localization analysis, wound-healing assays, and MTT assays. We further applied Connectivity Map (CMap) screening and molecular docking to identify candidate ULK3 activators. ULK3 was significantly upregulated in 13 cancer types, including Bladder Urothelial Carcinoma, Breast Invasive Carcinoma, and Lung Adenocarcinoma. In contrast, ULK3 was downregulated in Cholangiocarcinoma and Head and Neck Squamous Cell Carcinoma. High ULK3 expression was associated with poor overall survival in Adrenocortical Carcinoma, Kidney Renal Clear Cell Carcinoma, and Skin Cutaneous Melanoma. Copy number amplification contributed to ULK3 overexpression. A recurrent A206V missense mutation was detected in the protein kinase (Pkinase) domain. Genes co-expressed with ULK3 were enriched in RNA splicing, methylation, oxidative phosphorylation, and energy metabolism. ULK3 expression showed positive correlations with tumor stemness indices and m1A/m5C/m6A RNA modification regulators. From an immunological perspective, high ULK3 expression was associated with lower Immune Score, increased M2 macrophage infiltration, and co-expression of PD-L1, CTLA4, and LAG3 in most cancers. ULK3 expression was also correlated with Tumor Mutational Burden in Kidney Renal Clear Cell Carcinoma and Rectum Adenocarcinoma. In addition, ULK3 expression was associated with Microsatellite Instability in Brain Lower Grade Glioma, Lung Adenocarcinoma, and Uterine Corpus Endometrial Carcinoma. ULK3 overexpression promoted proliferation and migration in PC-3 cells. Cephaeline was screened as a putative ULK3 activator. Overall, ULK3 expression and amplification were associated with poor clinical outcomes, tumor stemness, immunosuppression, and RNA dysregulation. These findings highlight the potential value of ULK3 as a pan-cancer diagnostic and prognostic biomarker and as a predictor of immunotherapy response, particularly in prostate cancer. Full article

Nucleic acid amplification tests (NAATs) are central to modern virology diagnostics. However, evidence supporting alternative specimen types remains uneven across viral families, especially for emerging viruses. This limits diagnostic flexibility in outbreak and clinically complex settings. We conducted a scoping review of NAAT detectability across key body sites for human RNA viruses. PubMed and Embase were systematically searched for studies reporting NAAT results from urine, blood, fecal, cerebrospinal fluid, or respiratory specimens. Data were independently screened and synthesized to summarize specimen-specific detectability for each virus. From 8676 screened records, 321 studies were included, covering 39 viruses across 25 RNA virus families. Detectability across specimen types varied substantially between viruses. Consistent detection across multiple specimens was observed for few viruses, including SARS-CoV-2, Zika virus, and HIV, whereas many emerging viruses were evaluated in a single body compartment with limited comparative data. NAAT performance across specimen types is highly virus-specific and unevenly studied, with reliance on blood or respiratory specimens, potentially overlooking viable, less invasive alternatives. Evidence gaps are particularly pronounced for urine and cerebrospinal fluid, and heterogeneous reporting limits cross-study comparability. Standardized, cross-specimen and longitudinal studies are needed to improve diagnostic strategies, outbreak preparedness, and future assay development. Full article

Ovotransferrin (OVT) and lysozyme (LYS) are food-derived proteins with reported bioactive properties, but results from high-oil-fraction dispersions require cautious interpretation when phase type and undiluted particle size are not independently verified. This work screened a low-temperature OVT-LYS concentrated dispersion in which freeze-drying was omitted from the final preparation stage. LYS was first incorporated into a β-cyclodextrin/trehalose-protected LYS complex (LPC). A sequential one-factor-at-a-time design was used to screen hydrocolloid type, homogenization parameters, and flaxseed gum/xanthan gum mass ratio. Qualitative optical microscopy, zeta potential, creaming stability, pH/temperature tolerance, OVT immunoreactivity, and DLS data obtained after 1000-fold dilution were used as descriptors. Among the six tested hydrocolloids, flaxseed gum (FG) showed the most favorable overall single-hydrocolloid performance. High-pressure homogenization at 1000 bar for 1.5 min, corresponding to approximately three estimated equivalent passes, was selected as a practical processing condition within the tested range. A flaxseed gum:xanthan gum mass ratio of 1:2 showed the most favorable balance between pH/temperature tolerance and OVT immunoreactivity retention among the tested binary blends. Because LYS bioactivity assays, phase-type verification, DLS dilution-gradient validation, and undiluted particle-size measurement were not performed, the results support bounded formulation-screening evidence, rather than verified phase structure, retained LYS activity, or universal optimal parameters. Full article

The BRCA1 protein serves an essential function in maintaining genomic integrity, to the extent that up to 80% of women carrying a pathogenic BRCA1 variant develop breast cancer (BC). Most of these carriers would benefit from prophylactic care, but genetic screens that uncover variants of uncertain significance (VUSs) do not provide insight on disease risk or clinical decision-making. In accordance with guidelines established by The American College of Molecular Genetics (ACMG) and Association for Molecular Pathology (AMP), this study produced computational and functional evidence to inform the reclassification of BRCA1 VUSs as pathogenic or benign, with a specific focus on the abundant subset of missense variants within the RING domain. A six-feature linear support vector machine (LSVM) specifically trained on BRCA1 RING variants performed well (84% accurate in predicting in vitro binding loss) and provided supporting classification evidence for 322 VUS. A mammalian cell co-immunoprecipitation (co-IP) assay that quantified the binding between variant BRCA RING constructs and endogenous BARD1 provided corroborating strong evidence for nine VUSs and correlated with a homology-directed repair (HDR) assay by Starita et al. (p = 0.04). The combined evidence warrants the reclassification of three VUSs as likely benign (N16S, A17D, and E100D) and one as likely pathogenic (H41P), and underscores the promise of domain-specific approaches for missense VUS reclassification. Full article

Background/Objectives: Aggregatibacter actinomycetemcomitans is an important periodontal pathogen whose biofilm formation and bacterial aggregation contribute to colonization and persistence within the oral cavity. This study aimed to generate random transposon mutants of A. actinomycetemcomitans and screen for alterations in biofilm- and aggregation-related phenotypes. Methods: A transposon mutagenesis library was created in the serotype d strain SA269 using the EZ-Tn5 transposon system. Transformants were screened in vitro for biofilm formation, autoaggregation, and coaggregation with Fusobacterium nucleatum. Biofilm biomass was quantified using a standardized microtiter plate assay, while aggregation assays were performed by optical density measurements. Results: Among 44 transformants, 12 exhibited a markedly reduced ability to form biofilms compared with the wild-type strain, indicating that disruption of certain loci compromises surface adherence and biofilm biomass accumulation. Five representative mutants were selected for aggregation studies. Three mutants showed reduced autoaggregation, suggesting impaired intraspecies cell–cell interactions, whereas four mutants demonstrated enhanced coaggregation with F. nucleatum, reaching approximately 50% coaggregation after 120 min. These findings indicate that insertional inactivation of genes in A. actinomycetemcomitans can differentially affect biofilm formation, autoaggregation, and interspecies coaggregation. Conclusions: This preliminary study demonstrates the utility of random transposon mutagenesis for identifying mutants with altered adhesion- and biofilm-related phenotypes. Although the disrupted genes were not identified, the mutant library provides a foundation for future molecular characterization of genetic determinants involved in biofilm development and microbial community interactions. Full article

The inhibition of angiotensin-converting enzyme (ACE) to lower angiotensin is important in the treatment of hypertension (HT). ACE inhibitory activity is rarely documented in Thai traditional and indigenous medicine. Here, we evaluated the angiotensin I–converting enzyme inhibitory (ACEi) activity through bio-screening of selected medicinal plant species traditionally used for HT treatment by ethnic communities in northern Thailand, including Blumea balsamifera (L.) DC., Clerodendrum chinense (Osbeck) Mabb., Rotheca serrata (L.) Steane & Mabb., and Zingiber purpureum Roscoe. Using an in vitro assay, ethanolic extracts were evaluated for ACE inhibitory activity. Among the four extracts tested, the ethanolic leaf extract of Blumea balsamifera was the most effective by reducing ACE activity by 29, 36, and 64% at concentrations of 0.4, 2.0, and 10.0 mg/mL, respectively. The rhizome extract of Zingiber purpureum showed the second highest activity, with inhibition rates of 34%, 39%, and 40% at the corresponding concentrations. Cytotoxicity testing in HEK293T kidney cells was conducted to underscore the detectable toxicity under the tested conditions. Interestingly, intercultural and cross-cultural comparisons revealed a degree of agreement in the use of medicinal plants for hypertension treatment. Plant species traditionally used across multiple cultures tended to show higher levels of ACE inhibitory activity, suggesting their potential as candidates for the development of novel anti-hypertensive agents. To our knowledge, this is the first report describing the ACE inhibitory activity of medicinal plant species used for hypertension treatment by ethnic communities in northern Thailand. Full article

β-thalassemia is one of the most prevalent inherited hemoglobin disorders worldwide and represents a major public health challenge in Southeast Asia, particularly in Vietnam. The disorder is caused by pathogenic variants in the β-globin (HBB) gene, resulting in reduced or absent β-globin synthesis, ineffective erythropoiesis, chronic hemolytic anemia, and progressive multi-organ complications. Vietnam exhibits a high burden of thalassemia and related hemoglobinopathies, with approximately 13.8% of the population carrying a hemoglobinopathy-associated variant and substantial heterogeneity in distribution across ethnic groups and geographic regions. This review provides a comprehensive overview of the epidemiology, molecular characteristics, diagnostic strategies, treatment approaches, and prevention programs for β-thalassemia in Vietnam. Current evidence indicates that a limited number of recurrent HBB variants account for the majority of β-thalassemia alleles in the Vietnamese population, including codon 17 (A>T) (HBB: c.52A>T), codons 41/42 deletion (-TTCT) (HBB: c.126_129delTTCT), codons 71/72 (+A) (HBB: c.216_217insA), codons 95 (+A) (HBB: c.287_288insA), IVS-I-1 (G>T) (HBB: c.92+1G>T), IVS-I-5 (G>C) (HBB: c.92+5G>C), IVS-II-654 (G>T) (HBB: c.316+197C>T), -28 (A>G) (HBB: c.-78A>G), and -88 (C>T) (HBB: c.-138C>T). Diagnostic strategies generally follow a stepwise approach integrating hematological screening and hemoglobin analysis with confirmatory molecular testing. Advances in molecular diagnostics, particularly targeted polymerase chain reaction (PCR)-based assays and next-generation sequencing (NGS), have significantly improved detection of both carriers and affected individuals. Despite these advances, β-thalassemia continues to impose a considerable clinical, economic, and societal burden because many patients require lifelong blood transfusions, iron chelation therapy, and multidisciplinary management of disease-related complications. Major challenges include limited access to screening, prenatal diagnosis, and genetic counseling services, particularly in rural and ethnic minority populations, as well as the financial and technical barriers associated with advanced molecular diagnostics and emerging therapies. Strengthening nationwide screening programs, expanding access to prenatal and preconception genetic services, improving public awareness, and enhancing healthcare infrastructure are essential for reducing disease incidence and improving patient outcomes. In parallel, the integration of advanced molecular diagnostics and emerging gene-based therapies will be critical for advancing long-term disease control. This review highlights current knowledge gaps and proposes strategic priorities to support evidence-based policy development and comprehensive β-thalassemia prevention and management in Vietnam. Full article

Zanthoxylum bungeanum alkaloids have attracted considerable attention for their potential health benefits, yet systematic investigations into their extraction, purification, and comprehensive bioactivity remain scarce. In this study, an efficient extraction protocol was developed and scaled up, followed by purification using macroporous resin NKA-9 with 70% ethanol as the optimal eluent. Ultra-performance liquid chromatography coupled with ion mobility quadrupole time-of-flight mass spectrometry (UPLC-IM-QTOF/MS) identified 11 major alkaloids in the 70% ethanol eluate fraction. The antioxidant capacity of the 70% ethanol eluate fraction was evaluated through DPPH, ABTS⁺·, and Fe³⁺ reducing power assays, revealing a clear dose-dependent effect, albeit weaker than ascorbic acid. Antibacterial screening against four pathogenic bacteria (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa) via the Oxford cup method, macrobroth dilution, and minimum bactericidal concentration (MBC) determination demonstrated strain-selective activity. The strongest effect was observed against E. coli [minimum inhibitory concentration (MIC) = 2 mg/mL, MBC = 8 mg/mL, MBC/MIC = 4, bactericidal], followed by moderate activity against B. subtilis (MIC = 4 mg/mL, MBC = 16 mg/mL), while only bacteriostatic effects were noted against S. aureus and P. aeruginosa within the tested range. These findings provide a robust foundation for further development of Zanthoxylum bungeanum alkaloids as natural functional ingredients or food-compatible bio preservatives. Full article

Ilyonectria is a common soil-inhabiting fungal genus that comprises numerous plant phytopathogenic species capable of infecting a wide array of crops, medicinal herbs, and horticultural plants. However, the lack of a reliable and efficient genetic transformation method has severely hindered the elucidation of the pathogenic mechanisms of Ilyonectria pathogens. In this study, we established an efficient protoplast-mediated genetic transformation method for two dominant Panax root rot pathogens, I. robusta and I. vredehoekensis. Key parameters governing high-quality protoplast preparation, including mycelium culture time, enzyme composition, osmotic stabilizer type, digestion speed, and digestion time, were systematically optimized. Subsequently, orthogonal experiments were conducted to optimize the PEG-CaCl₂-mediated transformation conditions and to screen regeneration media for protoplasts. The optimal enzymatic system is composed of 20 mg/mL driselase and 10 mg/mL lysing enzyme, with 0.7 M NaCl as the osmotic stabilizer. Under these conditions, high-viability and high-quality protoplasts were obtained from I. vredehoekensis after 3 h of digestion at 150 rpm, and from I. robusta after 2 h of digestion at 100 rpm, yielding 5.52 × 10⁷ CFU/mL and 5.75 × 10⁷ CFU/mL protoplasts, respectively. Efficient transformation was achieved using a mannitol-prepared STC buffer mediated by 40% PEG4000. PCR and fluorescence microscopy verified positive transformants. Additionally, pathogenicity assays showed no significant differences in virulence between the transformed and wild-type strains, suggesting that the transformation procedure did not alter virulence. To the best of our knowledge, this is the first study to successfully establish genetic transformation methods for I. robusta and I. vredehoekensis, providing an essential technical platform for functional gene analysis, pathogenicity studies, and host–pathogen interaction research. In addition, the optimized transformation strategy may serve as a valuable reference for studies on other Ilyonectria species. Full article

The increasing prohibition of animal testing for cosmetic products has driven the development of alternative approaches to ensure consumer safety. Skin sensitization is one of the most critical toxicological endpoints to evaluate, requiring rigorous assessment to ensure the safety of cosmetic ingredients. This study proposes an Integrated Testing Strategy (ITS) that combines in chemico (Direct Peptide Reactivity Assay—DPRA) and in silico approaches (a six-platform computational panel) to evaluate the sensitization potential of substances. Initially, the in chemico methodology was validated through a partial proficiency demonstration to ensure experimental reliability. Subsequently, this ITS was applied to the Baccharis trimera extract and its major marker, 3-Caffeoylquinic acid (chlorogenic acid/3-CQA). Our results demonstrate that the DPRA alone is insufficient to classify the sensitization potential of complex mixtures, as recommended by OECD guidelines. The integration of in silico data proved essential to interpret the reactivity of the botanical matrix, revealing that the sensitization potential observed in the extract does not stem solely from 3-CQA, but likely results from the synergistic contribution of more lipophilic caffeoylquinic acid isomers. This approach demonstrates that integrating experimental and computational methods is fundamental for a robust safety assessment, offering an efficient, animal-free strategy for the early screening of cosmetic ingredients and for refining the interpretation of toxicological data in complex chemical environments. Full article

Saccharomyces cerevisiae FOSU-QQT (SC.QQT), isolated from pineapple pomace wine, exhibits favorable aroma-producing capabilities. In this study, we performed integrated genomic and phenotypic analyses to comprehensively evaluate its safety profile, probiotic potential, and aroma-producing characteristics. Whole-genome sequencing (WGS) assembly predicted a genome size of 30,256,254 bp, encompassing 12,899 genes with a total coding length of 22,062,659 bp and an average GC content of 37.30%. Preliminary safety assessments, including hemolysis tests, antibiotic susceptibility profiling, and antibacterial activity assays, were complemented by in silico screening for antibiotic resistance-associated genes. Functional tolerance assays, specifically resistance to simulated gastrointestinal fluid, acid stress, and bile salts, demonstrated that SC.QQT exhibited robust survival under physiologically relevant gastrointestinal conditions. Collectively, these findings support its potential as a promising probiotic candidate with notable resilience, although further in vivo validation is required to confirm its application value. Additionally, gas chromatography–mass spectrometry (GC–MS) analysis of volatile compounds in pineapple pomace wine indicated that the presence of aroma-related genes in SC.QQT may enhance overall flavor complexity and intensify fruity aromatic notes during fermentation, underscoring its distinctive utility in fruit wine bioprocessing. Full article

Background: Skin aging is driven by oxidative stress and ultraviolet (UV) exposure, leading to extracellular matrix degradation and loss of skin elasticity. This study aimed to identify the most biologically active Citrus species using a bioassay-guided approach and evaluate its potential for dermal applications. Methods: Hydroalcohol extracts and ethyl acetate fractions of Citrus sinensis, Citrus aurantifolia, and Citrus reticulata leaves were screened for antioxidant, enzyme-inhibitory, and polyphenol content. The most active fraction was characterized by UPLC-PDA and LC–MS/MS, formulated into Span-based nanovesicles, and evaluated for physicochemical properties and drug release. Biological activity was assessed using an in vitro scratch wound-healing assay on human dermal fibroblasts and a UVA-induced photoaging mouse model, supported by molecular dynamics simulations. Results: The ethyl acetate fraction of C. aurantifolia (CAE) exhibited the highest biological activity among the tested samples. This fraction showed potent antioxidant activity (DPPH IC₅₀ = 3.53 ± 0.05 µg/mL), marked inhibition of elastase (91.3%), collagenase (92.0%), and tyrosinase (80.2%), and a high total flavonoid content (110.49 mg rutin equivalents/g). Phytochemical profiling of CAE tentatively identified fourteen compounds, predominantly flavonoids, with hesperidin (30.4 mg/g) as a major constituent. The optimized nanovesicles (184 ± 0.9 nm, PDI 0.10, EE% 75.0%) enabled sustained hesperidin release. CAE and CAEnp enhanced fibroblast migration and accelerated wound closure at 24 h (p < 0.05). In vivo, CAEnp improved UVA-induced histopathological alterations and modulated oxidative stress-related markers by reducing p62/SQSTM1 by 28.7%, Keap1 expression to 21% compared with the CAE-treated group, and enhancing Nrf2, ARE, and NQO1 expression by 54.1%, 28.3%, and 57%, respectively. Molecular dynamics simulations supported stable hesperidin binding to elastase and suggested possible modulation of collagenase flexibility. Conclusions: The polyphenol-rich leaf fraction from C. aurantifolia, identified through comparative bioassay-guided fractionation, demonstrated antioxidant, enzyme-inhibitory, wound-healing, and photoprotective effects, particularly after nanoformulation. These findings support its potential for further development as a natural topical anti-aging candidate. Full article

Novel steroidal heterocyclic derivatives were prepared from cholesterol, stigmasterol, and diosgenin scaffolds via concise, reagent-efficient synthetic routes. Two 1,3-oxathiolan-5-one derivatives bearing cholest-5-ene (7) and stigmasta-5,22-diene (8) cores were obtained from the corresponding steroidal ketones through hydrazone formation, phenyl isothiocyanate addition, and S-selective cyclization with chloroacetic acid in refluxing toluene. An aminothiazole–diosgenin hybrid (12) was independently prepared via regioselective α-bromination of an oxidized diosgenin intermediate followed by condensation with thiourea in ethanolic sodium ethoxide. The newly synthesized target compounds were characterized by FT-IR, ¹H- and ¹³C-NMR spectroscopy, ESI mass spectrometry, and elemental (CHNS) analysis. The disc-diffusion assay against S. aureus and E. coli was retained only as a preliminary qualitative screen because no positive antibiotic control or MIC determination was included. Within this limited screen, compound 12 produced measurable 8.5–9.0 mm inhibition zones against both strains, supporting their prioritization for future standardized antimicrobial testing rather than establishing comparative potency. Full article

Marine medicinal algae represent a valuable reservoir of bioactive metabolites for anticancer drug discovery, yet the efficient identification of target-relevant compounds from chemically complex marine matrices remains challenging. In this study, an integrated cathepsin B-oriented strategy was developed to discover, prioritize, isolate, and validate antitumor metabolites from the brown alga Sargassum polycystum. Affinity ultrafiltration LC-MS was first applied to screen CTSB-binding constituents from the crude extract, followed by molecular docking, molecular dynamics simulation, and gray relational analysis for multidimensional candidate prioritization. Seven CTSB-binding metabolites were characterized, including chlorogenic acid, caffeic acid, cynarin, loliolide, taxifolin, senkyunolide H, and dihydroactinidiolide, with binding degrees of 73.99–85.61% at 2.5 U/mL CTSB. Molecular docking showed predicted binding affinities ranging from −6.3 to −9.4 kcal/mol, compared with −10.2 kcal/mol for the positive control CA-074Me. Integrated computational and biological evaluation identified caffeic acid, cynarin, and taxifolin as the top-ranked candidates. Preparative recovery was then achieved using counter-current chromatography combined with semi-preparative HPLC, and the isolated compounds were structurally identified by LC-MS/MS and NMR. Cellular assays in NCI-H1975 cells suggested that these metabolites reduced CTSB-associated enzymatic activity and intracellular CTSB-related fluorescence signals to different extents, with phenolic acid-type compounds exhibiting comparatively stronger effects. At the extract level, S. polycystum dose-dependently suppressed NCI-H1975 xenograft tumor growth, with inhibition rates of 48.78%, 36.58%, and 22.86% in the high-, middle-, and low-dose groups, respectively, without evident hepatorenal histopathological toxicity. This effect was associated with reduced CTSB, Ki-67, and Bcl-2 staining, increased Bax staining, enhanced apoptosis, and ultrastructural alterations in tumor tissues. Overall, this study provides a practical CTSB-oriented workflow for discovering antitumor metabolites from marine medicinal algae and supports further investigation of S. polycystum as a potential source of anti-NSCLC candidates. Full article

Isofenphos-methyl (IFP), a highly toxic and persistent organophosphate pesticide (OP), is widely used for soil pest control in crops but poses severe risks to ecological safety and human health because of its environmental accumulation and bioaccumulation. Herein, a sensitive and specific indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed for rapid IFP detection in vegetables. A novel IFP hapten was rationally designed and synthesized via computer-aided molecular simulation, and its structure was validated by liquid chromatography–tandem mass spectrometry (LC–MS/MS) and nuclear magnetic resonance (NMR). High-specificity anti-IFP monoclonal antibodies (mAbs) with strong anti-matrix interference were prepared for the first time using a matrix effect-enhanced screening strategy. The optimized ic-ELISA showed high sensitivity, with an IC₅₀ of 6.087 ng/mL and a detection range of 1.165–30.490 ng/mL, and no cross-reactivity with other common OPs. Spiked recovery experiments in celery and chili pepper matrices yielded recoveries of 81.87–97.95% (RSD < 5.44%), with highly consistent LC–MS/MS results. The method exhibited a weak positive matrix effect in vegetable matrices, eliminating complex pretreatment and enabling rapid onsite detection. Full article