Search Results (28,699)

Triple-negative breast cancer (TNBC) is characterized by prominent neutrophil infiltration; however, its significance remains controversial. Here, we investigated the role of neutrophil chemoattractant receptors in TNBC progression and metastasis. In contrast to wild-type (WT), Fpr^-/-, and Fpr2^-/- mice, neutrophils were almost completely absent in 4T1 tumors from Cxcr2^⁻/⁻ mice, indicating a dominant role for CXCR2 in the recruitment of tumor-associated neutrophils, leading us to use Cxcr2^-/- mice for further studies. Primary tumor growth was comparable between WT and Cxcr2^-/- mice, whereas lung metastasis was significantly increased in Cxcr2^-/- mice, with reduced expression of inflammatory cytokines, chemokines and cytotoxic molecules, including granzyme B and perforin, in primary tumors and metastatic lungs of Cxcr2^-/- mice. In vitro, WT, but not Cxcr2^-/-, neutrophils enhanced CD8⁺ T cell activation, partly via ICAM-1, and directly induced tumor cell death, supporting their anti-tumor function. To assess clinical relevance, transcriptomic data were analyzed. High neutrophil infiltration combined with elevated CXCR2 expression, and to a lesser extent CXCR1 expression, was associated with improved prognosis in patients with basal-like BC that largely overlaps with TNBC. Collectively, these findings suggest that CXCR2-mediated neutrophil recruitment exerts protective, anti-tumor effects and may represent a new prognostic marker for TNBC patients. Full article

Recent advances in enone chemistry have enabled the development of structurally optimized derivatives with improved anticancer selectivity. In this study, the cytotoxic activity and underlying mechanisms of sodium salts of four α,β-unsaturated enones (ES1–ES4), synthesized from vanillin-based scaffolds, were evaluated in human colorectal carcinoma (HCT-116), cervical adenocarcinoma (HeLa), and normal lung fibroblast (MRC-5) cell lines. All compounds exhibited concentration- and time-dependent cytotoxicity, with ES2 showing the highest potency (IC₅₀ = 14.25 μM in HCT-116 and 18.12 μM in HeLa at 72 h) and minimal toxicity toward MRC-5 cells (IC₅₀ > 90 μM). Although cisplatin demonstrated greater overall cytotoxicity, the enone salts displayed significantly higher selectivity indices, indicating a more favorable therapeutic window. Phase-contrast microscopy revealed characteristic morphological features of apoptosis, including cell rounding and membrane blebbing. Mechanistic investigations confirmed mitochondrial-mediated apoptosis, evidenced by increased early and late apoptotic populations, Bax upregulation, Bcl-2 downregulation, and caspase-3 activation. JC-10 staining demonstrated mitochondrial membrane depolarization accompanied by cytochrome c release. In addition, cell cycle analysis revealed pronounced G2/M phase arrest, particularly in HCT-116 cells. Collectively, these findings indicate that vanillin-derived enone sodium salts exert selective anticancer effects through mitochondrial apoptosis and cell cycle disruption, supporting their potential as low-toxicity anticancer candidates. Full article

Background: Cancer patients are highly susceptible to microbial infections due to immune suppression, necessitating therapeutic strategies that integrate anticancer efficacy with effective antimicrobial intervention. Chalcone-derived nitrogen-fused heterocycles represent a promising platform for developing multi-target agents with relevance to antimicrobial drug delivery, particularly for localized infections. Methods: A series of chalcone-based pyrazoline-thiadiazole nitrogen-fused azole hybrids was synthesized via thiosemicarbohydrazide-functionalized intermediates and fully characterized. Antiproliferative activity was evaluated against MCF-7, HepG-2, HeLa, and HCT-116 cell lines, alongside selectivity toward WI-38 normal fibroblasts. Antibacterial, antibiofilm, and in vivo efficacy were assessed against methicillin-resistant Staphylococcus aureus (MRSA USA300) and Acinetobacter baumannii AB5057. Mechanistic investigations included cell-cycle analysis, apoptosis assays, ERK2, RIPK3, p53, BAX/Bcl-2 quantification, DNA gyrase inhibition, molecular docking, molecular dynamics simulations, and density functional theory calculations. Results: Compound 13 exhibited potent cytotoxicity, particularly against MCF-7 (IC₅₀ = 3.87 ± 0.2 µM), outperforming doxorubicin (IC₅₀ = 4.17 ± 0.2 µM), with high selectivity indices (SI = 10.7 for MCF-7). Mechanistically, compound 13 induced G₂/M arrest (40.16% vs. 14.15% control), increased apoptosis to 32.89%, up-regulated ERK2 (3.17-fold), RIPK3 (11.97-fold), and p53 (3.54-fold), and markedly increased the BAX/Bcl-2 ratio (~42-fold). Compounds 7 and 13 displayed bactericidal activity against MRSA and A. baumannii (MIC/MBC = 10 mg/mL), potent antibiofilm effects, and significant in vivo efficacy in an MRSA skin infection model. Compound 13 reduced bacterial load by ~5 log units, outperforming vancomycin. DNA gyrase inhibition (IC₅₀ = 17.10 ± 0.17 µM) and computational studies supported target engagement. Conclusions: Pyrazoline-thiadiazole-based nitrogen-fused azole hybrids, particularly compound 13, demonstrated quantifiable anticancer and antimicrobial efficacy with strong in vivo validation, supporting their potential as multi-target candidates relevant to antimicrobial drug delivery in infection-prone cancer patients. Full article

Vascular endothelial growth factor (VEGF)-driven pathological angiogenesis constitutes a primary driver of neovascular diseases, including neovascular age-related macular degeneration (nAMD) and diabetic retinopathy (DR). Although anti-VEGF agents demonstrate clinical efficacy, their limited intraocular half-life mandates repeated intravitreal injections, thereby highlighting the imperative for long-term therapeutic strategies. In the present study, we assessed the anti-angiogenic potential of retinal organoid-derived microRNAs (miRNA) delivered via an engineered adeno-associated virus vector. Human umbilical vein endothelial cells (HUVEC) were transduced with AAV2.7m8 vectors to overexpress three candidate miRNA (miR-26a, miR-122, and let-7a), followed by VEGF stimulation to evaluate downstream signaling pathways and angiogenic responses. AAV2.7m8-mediated transduction of HUVEC demonstrated high efficiency without inducing detectable cytotoxicity. Overexpression of these miRNA markedly attenuated VEGF-induced phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK. Functional assays demonstrated suppression of endothelial cell proliferation and cell cycle progression, with miR-122-5p additionally inhibiting migration. All three miRNA substantially inhibited capillary-like tube formation. In aggregate, these results affirm that AAV2.7m8-mediated delivery of retinal organoid-derived miRNA —namely miR-26a-5p, miR-122-5p, and let-7a-5p—markedly suppresses VEGF-induced angiogenic signaling cascades and endothelial cell activation in vitro, thereby establishing their viability as a sustained therapeutic approach for pathological retinal neovascularization. Full article

Avibacterium paragallinarum (Av. paragallinarum), the causative agent of infectious coryza, imposes substantial economic burdens on the poultry industry by inducing growth retardation in broilers and reducing egg production in laying hens by up to 40%. Disease control is hindered by the limited efficacy of available vaccines and the increasing prevalence of antibiotic resistance—challenges that are exacerbated by the pathogen’s capacity to form biofilms, which facilitate bacterial persistence and enhance drug tolerance. To systematically elucidate the genetic determinants underlying biofilm formation in Av. Paragallinarum, we constructed a high-density random mutant library using mini-Tn5 transposon mutagenesis, comprising 3106 individual mutants. Phenotypic screening via crystal violet staining identified 188 mutants displaying altered biofilm-forming capacity relative to the wild-type strain, including 172 with enhanced and 16 with reduced biofilm formation. Sequencing of transposon insertion sites in these mutants revealed 105 disrupted genes involved in diverse biological pathways, including amino acid metabolism, quorum sensing, and transmembrane transport. A representative subset of eight mutants was selected for detailed phenotypic characterization. Their biofilm phenotypes were consistent with the initial screening results; certain mutants exhibited markedly enhanced biofilm formation (e.g., Tn-2206), whereas others, including Tn-1504, Tn-2428, and Tn-2859, showed significant reductions in biofilm production. Notably, these three biofilm-deficient mutants—harboring disruptions in a TonB-dependent receptor (Tn-1504), a GntP family permease (Tn-2428), and a hypothetical protein (Tn-2859)—displayed drastically attenuated virulence in vitro. Compared with the wild-type strain, these mutants exhibited reductions in cytotoxicity (up to 66.38%), cell adhesion (up to 50.68%), and invasive capacity, while maintaining normal growth kinetics. These findings indicate that the identified genes may play crucial roles in biofilm-associated virulence and highlight Tn-1504, Tn-2428, and Tn-2859 as promising candidates for the development of live attenuated vaccines. Collectively, this study provides a comprehensive genetic foundation for the rational design of novel anti-biofilm strategies against Av. paragallinarum. Full article

Eupatorium fortunei Turcz. (E. fortunei), a member of the Asteraceae family, is a widely utilized traditional medicinal herb in China. Historically, it has been employed to treat conditions such as influenza, nausea, anorexia, and various ailments associated with “pathogenic dampness”. To the best of our knowledge, this study presents the first systematic review of recent research on E. fortunei, based on a comprehensive literature search across both Chinese and international databases, including Web of Science, PubMed, SciFinder, and CNKI. The review encompasses its botanical characteristics, traditional applications, phytochemical composition, pharmacological properties, and toxicological profiles. Current research reveals a diverse array of phytochemicals in E. fortunei, with 162 compounds identified to date, including thymol derivatives, terpenoids, alkaloids, benzofurans, fatty acids, and other bioactive constituents. These compounds exhibit a broad spectrum of pharmacological activities, encompassing anti-cancer, anti-viral, anti-fungal, anti-inflammatory, and anti-diabetic effects. Among these, thymol derivatives and benzofurans emerge as the most prominent bioactive compounds, demonstrating potent cytotoxic effects against various tumor cell lines. Although E. fortunei is generally considered safe, certain pyrrolizidine alkaloids (PAs) present potential hepatotoxic risks, which can be mitigated through appropriate dosage control and formulation optimization. As a valuable traditional Chinese medicinal herb, E. fortunei exhibits substantial therapeutic potential. In conclusion, this review provides a comprehensive and systematic overview of current research on E. fortunei, offering scientific evidence and guidance for its rational development and clinical application. Full article

Endophytic fungi associated with date fruits (Phoenix dactylifera) are mostly under-explored, despite their potential as reservoirs of natural compounds. The aims of this study were to characterize the endophytic fungus Aspergillus terreus (SU5) isolated from date fruits, and to investigate its biological activities and chemical profile for the first time. Morphological and molecular methods were utilized to identify Aspergillus terreus. A liquid chromatography–mass spectrometry analysis (LC/MS/MS) was conducted to determine the chemical profile of the crude extract. Biological properties were investigated through acetylcholine esterase and butyrylcholine esterase inhibition, cytotoxicity assays against MCF-7 and MCF-7/Adr, and antioxidant assays. LC/MS/MS of the fungal extract resulted in the detection of 39 of established secondary metabolites, primarily comprising polyketides, quinones, and phenolic derivatives. The crude extract demonstrated significant antioxidant activity, especially in the ABTS assay (IC₅₀ = 50.18 μg/mL), considerable cytotoxicity against MCF-7 breast cancer cells, diminished efficacy against the drug-resistant MCF-7/Adr cell line, and preferential inhibition of butyrylcholinesterase compared to acetylcholinesterase. While none of the identified compounds are novel, numerous metabolites are documented here for the first time from an endophytic A. terreus associated with date fruits. The findings underscore date fruits as a prospective ecological niche for a chemically varied endophytic fungus with potential pharmaceutical significance. Full article

Background: Iron deficiency remains a major nutritional challenge, partly due to the limited stability and bioavailability of conventional iron formulations in foods and during digestion. In this study, iron–protein microparticles (IP-MPs) based on bovine serum albumin (IA-MPs) and hemp protein (IH-MPs) were developed via coprecipitation and evaluated as food-compatible iron delivery systems. Methods: Iron–protein microparticles (IP-MPs) were fabricated by a coprecipitation technique. The stability of IP-MPs was investigated in a three-phase digestion model. The uptake of IP-MPs by Caco-2 cells as well as the Ferritin concentration in Caco-2 cells were investigated. Results: Particle morphology and size distribution were strongly dependent on the protein matrix, with hemp protein microparticles exhibiting greater size uniformity and higher stability under simulated gastric conditions. In a standardized in vitro gastrointestinal digestion model, both IP-MP formulations preserved iron predominantly in the bioactive Fe(II) state and remained sufficiently intact to reach the intestinal phase. Biocompatibility and iron uptake were assessed using Caco-2 cell monolayers. Neither formulation induced cytotoxic effects, while iron delivered via IP-MPs showed enhanced cellular uptake compared to a commercial iron supplement and ferrous sulfate. The amount of Fe(II) detected in the basolateral compartment of IH-MP and IA-MP samples (1.4 µg and 1.3 µg, respectively) was higher than that observed for Floradix^® samples (approximately 0.7 µg) and corresponded to about 25% of the total iron applied. Functional iron bioavailability, assessed by ferritin formation, was significantly higher for IP-MPs, with hemp protein microparticles yielding the strongest ferritin response. Conclusions: These results demonstrate that iron–protein microparticles, particularly those based on hemp protein, effectively improve iron stability during digestion and enhance cellular iron bioavailability, highlighting their potential for application in iron fortification and functional food systems. Full article

Plant-derived saponins have attracted significant interest for their potential to promote apoptotic cell death and enhance antitumor immune responses through immunogenic cell death (ICD). Akebia quinata, a saponin-rich medicinal plant, exhibits diverse pharmacological properties; however, studies on its seeds are limited, and their immunomodulatory activity in cancer remains largely unexplored. In this study, A. quinata seeds were extracted using 70% ethanol, and the phytochemical profile was characterized using UHPLC–QTOF MS/MS. We investigated the anticancer properties of A. quinata seed extract (AQSE), focusing on its role in inducing apoptosis and ICD in non-small cell lung cancer (NSCLC). In human NSCLC cell lines (A549 and H460), AQSE exhibited potent cytotoxic effects in a dose-dependent manner. Flow cytometric analysis confirmed the induction of apoptosis, evidenced by a significant increase in Annexin V-positive cells and an elevated sub-G1 population. Mechanistically, AQSE treatment induced cell death by simultaneously inhibiting the survival-promoting MEK/ERK/CREB axis and activating the stress-responsive JNK pathway. Furthermore, AQSE triggered hallmark features of ICD, characterized by surface exposure of calreticulin and the release of extracellular HMGB1 and ATP. Most importantly, an in vivo vaccination assay using a syngeneic mouse model demonstrated that immunization with AQSE-treated dying cells significantly suppressed tumor growth upon rechallenge, confirming the establishment of antitumor immunological memory. Additionally, bioassay-guided fractionation revealed that the anticancer activity was primarily concentrated in the ethyl acetate fraction. These findings suggest that AQSE exerts anticancer effects via the induction of apoptosis and ICD, highlighting its potential as a promising natural candidate for the development of novel therapeutic strategies against NSCLC. Full article

Cell-penetrating peptide (CPP) conjugation represents a promising strategy for enhancing the biological activity of therapeutic peptides. In this study, three analogues were designed by conjugating the trypsin inhibitory loop (TIL) derived from a Bowman–Birk-type inhibitor with the transactivator of transcription (TAT) peptide to improve their bioactivity. All TAT-TIL conjugates exhibited significantly enhanced antimicrobial activity compared with the parent peptide. Notably, the analogue containing a glycine linker (-GG-) showed further improvement in antiproliferative activity against cancer cells, indicating the potential role of linker design in optimizing peptide function. All analogues exhibited low hemolytic activity at the highest tested concentrations, although increased cytotoxicity toward normal HaCaT cells was observed, suggesting the need for further optimization of selectivity. Interestingly, comparable antimicrobial activities were observed regardless of protease inhibitory capacity, indicating that protease inhibition is not essential for the enhanced biological effects. Overall, TAT conjugation significantly improves the biological activity of Bowman–Birk-type inhibitor-derived peptides, and the incorporation of a glycine linker further enhances their functional properties. These findings support CPP-mediated peptide modification as an effective strategy for developing potential antimicrobial and anticancer peptide candidates. Full article

Glucagon-like peptide-1 (GLP-1) is an incretin hormone secreted by intestinal endocrine L cells that activates the GLP-1 receptor (GLP-1R), leading to glucose-dependent insulin secretion and suppression of glucagon release. In recent years, GLP-1R agonists (GLP-1RAs) have become one of the leading therapeutic options for the treatment of type 2 diabetes mellitus; however, for a long time clinically approved GLP-1RAs were limited to peptide drugs unsuitable for oral administration. The discovery of the “first-in-class” small molecule agonist danuglipron in 2018 demonstrated the feasibility of orally available GLP-1RAs and stimulated the development of numerous danuglipron-like compounds, some of which showed increased efficacy over the prototype. In this study, we report the design and synthesis of novel GLP-1RAs based on a regioisomeric danuglipron scaffold, 1H-benzo[d]imidazole-5-carboxylic acid. A series of 35 compounds was synthesized and evaluated in vitro for cytotoxicity and GLP-1R agonistic activity using a cAMP accumulation assay. A potent lead compound 12r (pEC₅₀ = 7.72, pCC₅₀ < 3.60) was found which is a close structural analog of danuglipron with reduced cytotoxicity and excellent selectivity over two other class B GPCRs, including GCGR and GIPR. Despite decreased potency compared to danuglipron, the obtained results hold promise for further optimization and provide valuable structure–activity relationship insights. Full article

Introduction: Spondyloenchondrodysplasia with immune dysregulation (SPENCDI) is a rare autosomal recessive disorder caused by biallelic variants in the tartrate-resistant acid phosphatase 5 (ACP5) and characterized by variable skeletal, immunological, and neurological manifestations. Because early skeletal abnormalities may be subtle, diagnosis can be challenging in infancy. Materials and methods: We conducted a detailed clinical, immunological, radiological, and molecular evaluation of an infant with early-onset cytopenia, recurrent infections, seizures, and developmental delay. Genomic analysis was performed using whole exome sequencing (WES) and copy number variation sequencing (CNV-seq). In addition, we performed a structured narrative review of published ACP5-related SPENCDI cases to summarize the clinical spectrum and the currently reported use of Janus kinase (JAK) inhibitors. Results: Genomic analysis identified an ACP5 stop-gain variant (c.311G>A; p.Trp104*) with an apparently homozygous signal on WES. Re-evaluation of the copy-number data demonstrated an overlapping heterozygous 19p13.2–p13.13 deletion encompassing ACP5, indicating biallelic ACP5 defects consisting of a sequence variant on one allele and deletion of the other allele. Clinically, the patient showed prominent extra-osseous manifestations, including impaired T- and NK-cell cytotoxicity, before the emergence of definite radiographic skeletal abnormalities. Our literature review showed that skeletal abnormalities were repeatedly documented across published ACP5-related SPENCDI reports, although radiographic changes were often subtle and could be preceded by immune manifestations. Reported use of JAK inhibitors suggests potential benefit for immune dysregulation in selected patients, whereas the neurological response remains uncertain. Conclusions: This study reports a novel ACP5 variant and expands the known phenotypic spectrum of SPENCDI. SPENCDI should be considered in children with unexplained immune dysfunction and developmental delay, and suggestive neuroimaging findings, even when overt skeletal deformities are absent. Early genetic testing and targeted skeletal imaging may facilitate diagnosis. Full article

Background: Photodynamic therapy (PDT) offers a promising complementary strategy for treating glioblastoma multiforme (GBM); however, limited control over photosensitizer activation and reduced efficacy under hypoxic conditions remain significant limitations. Methods: In this study, we present the synthesis and functional evaluation of Gal-SiX, an enzymatically activatable Si-xanthene-based activatable PDT agent designed to address these challenges. Prepared via an improved 10-step synthetic route, Gal-SiX exhibits clear turn-on fluorescence and absorbance responses upon β-galactosidase activation and efficiently generates reactive oxygen species in aqueous media. Results: Mechanistic studies revealed that Gal-SiX enables both Type I and Type II PDT pathways, a favorable feature for GBM environments characterized by restricted oxygen availability. In vitro assays conducted on U87MG glioblastoma cells and L929 healthy fibroblasts demonstrated light-dependent cytotoxicity, with IC₅₀ values of 3.30 μM and 7.19 μM, respectively. Gal-SiX also showed minimal dark toxicity (>80 μM) and potent light-induced cytotoxicity, yielding a phototoxicity index of 24.8 in glioblastoma cells. Confocal imaging and MTT assays consistently confirmed enzymatic activation and effective PDT response at the cellular level. Conclusions: Overall, this work introduces the first activatable Si-xanthene-based PDT agent for glioblastoma and provides the first evidence that the Si-xanthene scaffold can support dual Type I/II phototoxicity. These results underscore Gal-SiX’s potential as a PDT platform for addressing the unique constraints of GBM biology. Full article

Background: Colorectal cancer (CRC) continues to represent one of the most common and lethal malignant tumors globally. Notably, only patients diagnosed with microsatellite instability-high (MSI-H) colorectal cancer derive substantial clinical benefits from immune checkpoint inhibitor therapy. As critical immune cells that infiltrate tumors, γδT cells are tightly linked to the therapeutic response in colorectal cancer patients with microsatellite instability (MSI) colorectal cancer. However, the heterogeneous characteristics of γδT cells in colorectal cancer with different microsatellite statuses and their specific roles in regulating immunotherapy responses remain unclear. Methods: We performed dimensionality reduction and clustering analysis on γδT cells from a single-cell RNA sequencing dataset to explore diversity and functional characteristics of distinct γδT cell subsets. Meanwhile, bulk transcriptome data were applied to further investigate the immune infiltration, clinical characteristics, and immune checkpoint molecule expression in CRC patients stratified by distinct γδT cell subpopulations. Results: We identified five γδT cell subsets, among which the C4_CXCL13 γδT cell subsets was enriched in MSI CRC and exhibited an exhausted-like T cell phenotype while retaining robust cytotoxic function. A signature score based on these 17 marker genes was associated with survival, immune infiltration, and therapeutic response, thus representing a potentially valuable independent prognostic factor. Conclusions: The C4_CXCL13 γδT cell subset represents a characteristic subset in MSI CRC and is closely associated with clinical prognosis and benefit from immunotherapy. It represents a potential clinical marker for classifying patients and estimating the response to immunotherapy, offering a novel target for personalized immunotherapy in CRC. Full article

Background/Objectives: The effects of silver-coated dressing on wound healing, including cytotoxicity, are controversial due to the limited and incongruous results of in vitro versus in vivo research. Multiple factors intervene in wound healing processes and scarring, including pro/anti-inflammatory and pro/anti-fibrosis markers. Herein, to elucidate reported differences between in vitro and in vivo results, the effects of silver-coated dressing on 2D and 3D mono- and cocultures of fibroblasts and adipose-derived stem cells (ADSC) were investigated. Methods: Migration profiles in 2D and 3D assays, α-smooth muscle actin and proliferation marker Ki-67 expression, TGF-β1, TGF-β3, IL-6 and IL-10 levels and/or gene expression were assessed on four culture constructs. Results: In 2D systems at 24 h, silver-treated ADSC monocultures displayed better migration abilities compared to cocultures with high fibroblast ratio. In contrast, changes in the sprouting pattern between treated and untreated samples were non-significant in 3D constructs. TGFβ-1 levels decreased post-treatment, while TGFβ-3 increased, especially in 3D models. IL-6 gene expression was up-regulated following silver exposure in 3D models, mainly for stem cells in mono- and cocultures. Conclusions: Experiment data on 3D constructs suggest that silver-coated dressings do not significant impede wound healing, whereas cytotoxic effects were more pronounced in the 2D cultures. These inconsistencies, also noted in the literature, invite a methodological discussion of the 2D setup implications, recommending 3D constructs as a more appropriate evaluation standard where observable effects are closer to in vivo conditions and more relevant for transfer to clinical applications. Full article