Small extracellular vesicles (sEVs) are pivotal mediators of intercellular epigenetic communication in cancer. Following MISEV2023, we use the size- and isolation-based term sEV throughout, because most primary studies cited here cannot resolve which biogenetic route generated the vesicles they analyzed. By selectively packaging and transferring noncoding RNAs (ncRNAs), DNA fragments, chromatin-modifying enzymes, and metabolic effectors, sEVs reprogram recipient-cell chromatin architecture without altering the underlying DNA sequence. Tumor-derived sEVs engage stromal, immune, and vascular compartments to drive malignant progression through mechanisms that include miRNA-directed suppression of DNA methyltransferases (DNMTs), lncRNA-scaffolded Polycomb Repressive Complex 2 (PRC2) recruitment, depositing H3K27me3, and oncometabolite-mediated inhibition of TET dioxygenases. This narrative review synthesizes mechanistic, preclinical, and translational evidence on sEV-driven epigenetic regulation in cancer, applies a four-level evidence hierarchy to calibrate mechanistic claims, and critically evaluates how distinct cargo classes–microRNAs (miRNAs), long noncoding RNAs (lncRNAs), circular RNAs (circRNAs), DNMTs, and histone-modifying enzymes–contribute to chromatin remodeling, aberrant DNA methylation, acquired therapy resistance, and immune evasion in recipient cells. We further examine sEV cargo signatures as minimally invasive liquid biopsy biomarkers and appraise engineered sEV platforms for the precision delivery of miRNA mimics, siRNAs, and small-molecule epigenetic inhibitors. Key methodological challenges, EV isolation standardization, MISEV2023 compliance, cargo stoichiometry at physiological concentrations, in vivo biodistribution, and the transition from post-transcriptional regulation to durable chromatin-state change are critically evaluated, and a translational roadmap is proposed to guide reproducible clinical implementation of sEV-mediated epigenetic cancer therapeutics.
Microglia are key regulators of central nervous system homeostasis and neuroinflammation, and cannabinoid type 2 receptors (CB2Rs) have emerged as important modulators of microglial function. Although the pharmacology of CB2Rs has been extensively characterised in heterologous expression systems and activated microglia, comparatively little is known about the behaviour of CB2R ligands in non-stimulated microglia. The present study therefore aimed to characterise the pharmacological properties of a panel of CB2R ligands in non-stimulated BV-2 microglial cells and to determine whether constitutive receptor activity and ligand-dependent signalling bias could be detected under basal conditions. Classical CB2R agonists (CP 55,940, WIN 55,212-2, JWH 133 and JWH 015), putative protean agonists ((R)-AM 1241 and GW 405833), and inverse agonists (SR 144528, AM 630 and JTE 907) were evaluated using [35S]GTPγS binding and forskolin-stimulated cAMP assays. In the [35S]GTPγS assay, WIN 55,212-2 displayed the highest intrinsic activity, whereas (R)-AM 1241 and GW 405833 behaved as partial agonists. Inverse agonists reduced basal signalling, indicating constitutive CB2R activity in resting BV-2 cells. In contrast, cAMP measurements revealed greater signal amplification, with (R)-AM 1241 and GW 405833 exhibiting full agonist behaviour and SR 144528 producing pronounced inverse agonism. Marked differences in ligand efficacy and rank order between assays highlighted the influence of downstream signalling mechanisms and ligand-dependent signalling bias. Notably, GW 405833 displayed a strongly biased signalling profile, whereas SR 144528 consistently exhibited the greatest inverse agonist activity. These findings demonstrate that CB2Rs are functionally active in non-stimulated microglia and that constitutive receptor activity and signalling bias contribute significantly to their pharmacological profile under basal conditions. By focusing on non-stimulated microglia, this study provides new insights into CB2R signalling in a homeostatic cellular environment and establishes a framework for understanding how CB2R pharmacology may be altered during neuroinflammatory and neurodegenerative disease states.
Metal-graphitic nanocapsules are an emerging class of metal-graphitic hybrid nanomaterials, typically consisting of a metal core confined within a single- or few-layer graphitic shell. This unique core–shell architecture integrates the tunable physicochemical properties of metal nanomaterials with the chemical stability, Raman activity, fluorescence-quenching capability, and surface functionalization capacity of graphitic materials. In particular, the metal core can provide plasmonic enhancement as well as magnetic or catalytic auxiliary functions, while the chemically protective graphitic shell protects the core from harsh environments and provides intrinsic Raman bands that can serve as internal standards under well-controlled conditions. These features make metal-graphitic nanocapsules highly attractive as robust nanoprobes for molecular spectroscopy-based chemical analysis, biosensing, and targeted diagnosis. In this review, we first summarize the synthesis strategies, formation mechanisms, and key properties of representative metal-graphitic nanocapsules. We then discuss recent advances in their use across representative analytical and biomedical scenarios, with emphasis on the integration of spectroscopic readouts with targeted recognition strategies. Particular attention is given to how the metal core and graphitic shell cooperatively enhance signal generation, molecular enrichment, selective recognition, environmental stability, internal calibration, and reliable in situ diagnosis in real samples and living systems. Finally, we discuss current challenges and future perspectives for developing metal-graphitic nanocapsules as versatile platforms for molecular spectroscopy-based analysis and diagnosis.
Circadian rhythms determine biological clocks that manage daily biological functions, including the sleep–wake rhythm, hormone release, and immune function. This can affect immune-mediated chronic inflammation in rheumatoid arthritis (RA), which leads to joint damage, pain, swelling, and morning stiffness. These clinical manifestations follow a distinct circadian rhythm because of the cyclic pattern of leukocyte movement, secretion of pro-inflammatory cytokines such as IL-6, IL-1β, and TNF-α at night, and fluctuations in cortisol and melatonin levels. Despite the increasing amount of evidence showing that insufficient sleep and disruption of the biological clock may play a role in RA development through suppression of the immune response in macrophages and resorption processes in osteoclasts, the cause of this phenomenon is still under investigation. The present-day guidelines continue to endorse DMARDs as the basis of treatment, while studies are exploring the role of chronotherapy as an adjunctive method for alleviating symptoms and enhancing disease outcomes. In this review, we explore current knowledge regarding the links between the molecular basis of circadian biology and the pathogenesis of RA, highlight recent trends in immune and bone remodeling mechanisms, and discuss modern possibilities for using chronotherapy, precision medicine, and advanced drug-delivery methods based on circadian rhythms to create optimal treatment strategies for RA.
Breast cancer is among the most common cancers globally. While several advancements have been made to improve breast cancer management, there is still a need to find innovative ways to deliver drug therapy to improve efficacy and side effects associated with treatment. Among these methods, local and targeted drug delivery is particularly promising. The current review highlights localized polymer-based methods to effectively deliver breast cancer therapy, with a specific focus on the strengths and limitations of these systems. Injectable and surgically implanted scaffolds, microneedles, topical patches, liquid and semisolid topical drug carriers are amongst some of the delivery systems discussed. Highlighted in the discussion is how physiological changes that occur during breast cancer should be considered and utilized when developing drug formulations, by specifically exploiting the tumor microenvironment. Remaining gaps and future areas for drug delivery research are highlighted including personalized medicine and insights into novel drug delivery systems like nanomedicines and three-dimensional drug printing.
Small open reading frames (sORFs) encode micropeptides, which are a promising yet largely untapped resource for creating peptide design templates. Owing to their concise nature and functional efficiency, micropeptides often rely on essential structural elements and brief linear motifs, such as domains for membrane interaction, targeting sequences, and sites for protein–protein interactions, to fulfill their biological functions. This inherent simplicity makes them particularly suitable for a bottom-up design approach aimed at identifying, extracting, and systematically refining functional motifs to develop novel bioactive peptides. This review addresses the critical question of how micropeptides, particularly those involved in tumor regulation, can be explored as emerging therapeutic targets, functional templates for peptide design, and potential future therapeutic agents, by synthesizing current understanding of their mechanisms, functional significance in cancer, and the computational and design strategies for their clinical translation. We examined the current methods for analyzing the sequence and structural characteristics that underpin their functional activity and investigated how these attributes can be leveraged for drug discovery and design. Finally, we underscore the primary challenges and future prospects in converting sORF-encoded micropeptides into clinically relevant molecules with the aim of broadening the current scope of the druggable proteome.
Gambling disorder (GD) constitutes a worldwide social and economic burden and is associated with impaired functioning and reduced quality of life. GD shares important mechanistic substrates with obsessive–compulsive disorder (OCD), including dysfunction of cortico-striato-thalamo-cortical circuitry and dysregulation of serotonergic pathways involved in impulsivity, compulsivity, and impaired inhibitory control. On this basis, selective serotonin reuptake inhibitors (SSRIs), widely used in several psychiatric disorders, have been investigated as potential pharmacological treatments for GD. Evidence concerning fluoxetine, fluvoxamine, paroxetine, sertraline, citalopram, and escitalopram is heterogeneous and overall limited. Some early single-blind, randomized, and open-label studies have reported reductions in gambling urges, severity, and compulsive symptoms. However, larger and more rigorous placebo-controlled trials have frequently failed to demonstrate consistent superiority over placebo. Interpretation of these findings is further limited by small sample sizes, short observation periods, high dropout rates, heterogeneous outcome measures, and substantial placebo response. While SSRIs remain biologically plausible candidates for modulating the compulsive and impulsive dimensions of GD, current evidence does not support their routine use as first-line pharmacological treatment. Their role appears most justified in the presence of psychiatric comorbidity or within individualized, phenotype-oriented treatment strategies.
Accurate analysis of prostate cancer (PC)-related biomarkers requires sensing platforms capable of sensitive and multiplex detection in complex biological environments. Herein, we propose a signal-on electrochemical aptamer-based sensor (E-AB) for the simultaneous detection of L-lactate (L-Lac) and prostate-specific antigen (PSA). To maximize analytical performance, two Lac aptamer sensing configurations, single-stranded (ssLac201) and double-stranded (dsLac201), were constructed and comparatively evaluated. The dsLac201 structure displayed more effective background suppression and enhanced target induced signal response. Under optimized conditions, the dsLac201-based sensor exhibited a wide linear range from 500 nM to 10 mM for L-Lac, with a low detection limit of 157 nM and high selectivity. Based on this optimized design, a dual-aptamer electrochemical platform was further engineered through programmable nucleic acid assembly, enabling simultaneous detection of L-Lac and PSA via dual-input signal integration. The dual-target sensor showed broad analytical ranges for both biomarkers (L-Lac: 500 nM–10 mM; PSA: 10 pg mL−1–500 ng mL−1) and retained promising performance in serum samples. This work demonstrates a simple and versatile strategy for multiplex electrochemical biosensing and provides a promising platform for PC-related biomarker monitoring and clinical biomedical analysis.
Supportive care is essential during chemotherapy for head and neck cancer, yet the role of traditional Japanese medicine (kampo) remains unclear; therefore, we investigated whether Ninjin’yoeito (NYT) could reduce adverse events during cisplatin-based chemotherapy. We retrospectively analyzed 47 patients treated between June 2022 and June 2024, dividing them into an NYT group and a control group. Hematological toxicities, including decreases in white blood cells, neutrophils, hemoglobin, and platelets, as well as gastrointestinal disorders such as nausea, were evaluated. Compared with controls, patients receiving NYT showed significantly lower incidences of decreased white blood cell counts (p = 0.04), decreased hemoglobin levels (p = 0.03), and gastrointestinal disorders (p = 0.04). Trends toward reduced neutropenia and thrombocytopenia were also observed, although these did not reach statistical significance. These findings suggest that NYT may help mitigate hematological and gastrointestinal toxicities associated with cisplatin-based chemotherapy in patients with head and neck cancer. However, given the retrospective design and limited sample size, prospective studies are needed to confirm the efficacy and safety of NYT in this setting.
Hepatocellular carcinoma (HCC) is a leading cause of cancer mortality, frequently arising from chronic inflammatory states such as metabolic dysfunction-associated steatotic liver disease and cirrhosis. While extensive epidemiological data demonstrate a strong, dose-dependent inverse association between habitual coffee consumption and HCC incidence, the underlying molecular causality remains incompletely understood. In this comprehensive review, we elucidate the “Coffee Paradox” through the lens of nutriepigenomics. We demonstrate how coffee-derived bioactives—specifically chlorogenic acids, diterpenes, and microbially derived short-chain fatty acids—function as a coordinated epigenetic defense system. These compounds actively inhibit DNA methyltransferases, serve as endogenous histone deacetylase inhibitors via the gut–liver axis, and induce post-transcriptional, tumor-suppressive microRNA networks to halt oncogenic progression. However, to provide a critical and balanced perspective, we also address significant translational challenges. We evaluate conflicting null associations from recent Mendelian randomization studies and highlight the profound variability introduced by specific brewing methods, roasting profiles, and individual pharmacogenomics (e.g., CYP1A2 polymorphisms). Finally, we outline the future of precision hepatology, emphasizing the critical transition from observational epidemiology to clinical application via the utilization of circulating exosomal microRNAs as dynamic liquid biopsies and the development of standardized epi-nutraceuticals. Ultimately, this multi-layered epigenetic framework provides a robust foundation for integrating targeted dietary interventions into the primary prevention of HCC.
Intracranial mesenchymal tumors (IMTs) with FET::CREB fusion are rare mesenchymal neoplasms that rely on the confirmation of the molecular hallmark FET::CREB gene fusion for diagnosis. We report a case of a 53-year-old female presenting with neurocognitive decline and seizures. Neuroimaging demonstrated a heterogeneously enhancing solid-cystic lesion in the left frontal lobe. Gross total resection (GTR) of the tumor was achieved and the patient recovered to premorbid status. Definitive diagnosis was achieved via next-generation sequencing that identified an EWSR1 (exon 7)::CREM (exon 7) fusion transcript. A systematic literature review of 72 IMTs with FET::CREB-positive cases was performed in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 guidelines. Publications reporting confirmed FET::CREB fusion-positive IMTs, without restriction on year of publication, were included to analyze clinicopathological correlations and prognostic determinants. Mean age at diagnosis was 27.8 (±18.3). Patients who underwent GTR demonstrated a significantly lower rate of recurrence compared to those who underwent subtotal resection (STR) (p < 0.001), suggesting that extent of resection may be an important prognostic factor; however, causal inference is precluded by the observational nature of the data. Patients who received adjuvant therapy had a higher rate of recurrence (p = 0.043); however, this association is likely attributable to confounding by indication, as adjuvant treatment was predominantly administered to patients with subtotal resection or more aggressive disease. No causal inference regarding adjuvant therapy efficacy can be drawn from these data. Our study results corroborate that accurate diagnosis relies on molecular interrogation and the extent of resection appears to be an important prognostic factor for IMTs with FET::CREB fusion.
Hepatic hemangiomas are the most common benign liver tumors and are typically small and asymptomatic; however, pedunculated and exophytic variants are extremely rare and may mimic extrahepatic lesions on imaging, posing a potential diagnostic challenge. The aim of this study was to describe the multimodal imaging features of a pedunculated hepatic hemangioma arising from the left triangular ligament and to review the available literature with particular attention to MRI findings and diagnostic considerations. A 52-year-old man underwent contrast-enhanced thoracoabdominal CT for unrelated symptoms, which incidentally revealed a pedunculated hepatic lesion. Further evaluation was performed with multiparametric MRI at 1.5T, including diffusion-weighted imaging and dynamic contrast-enhanced sequences. A review of the English-language literature published up to 2025 focusing on pedunculated and exophytic hepatic hemangiomas was also conducted. CT and MRI demonstrated imaging features consistent with hepatic hemangioma, including peripheral nodular enhancement with progressive centripetal fill-in and marked T2 hyperintensity. Multiplanar MRI depicted a thin vascular pedicle connecting the lesion to the hepatic capsule, supporting its hepatic origin. Fewer than approximately 30 well-documented cases have been reported in the English literature. Recognition of these imaging findings may facilitate correct diagnosis and help avoid unnecessary invasive procedures.
Cerebral amyloid angiopathy (CAA) is a prevalent cerebral small vessel disease (CSVD) and prominent cause of vascular contributions to cognitive impairments and dementia (VCID). CAA is characterized by progressive accumulation of fibrillar amyloid β in cerebral vessel walls, leading to vascular wall degeneration, thrombotic occlusions, microbleeds, and perivascular inflammation causing cognitive deficits. Underlying mechanisms of CAA progression are poorly understood, and there exist no validated diagnostic biomarkers or therapeutic targets for CAA. Here, we performed proteomic mass spectrometry analysis of whole brain tissue and cerebral microvessel enriched fractions from the rTg-DI rat, a validated and well-characterized preclinical rat model of CAA, to identify potential targets and diagnostic markers related to vasculopathy progression in CAA. There were 92 increased and 104 decreased proteins identified in the rTg-DI whole brain samples, while 29 proteins were identified as increased in the enriched cerebral microvessel fractions. We identified several significant differentially expressed proteins in both sample types with commonly elevated proteins including HTRA1, APOE, APOD, CLU, MDK, and LAP3. This study also confirmed the differential expression of several proteins previously reported in isolated brain regions of rTg-DI rats, including ANXA3, APOD, APOE, CLU, CST3, CTSS, CTSD, CTSZ, GFAP, GSTA3, HTRA1, and ITGB2, with APOE, CLU, HTRA1, and GPC1 all reported in human CAA cases. We thus provide a “high-confidence” list of potential marker candidates from proteomic analysis in the rTg-DI rat model of capillary CAA type-1.
Composed of a steroid nucleus, widely distributed in the animal and plant kingdoms, containing various hydroxyl and methyl groups, and a carboxyl side chain, bile acids (BAs) appear to be the result of an irreversible evolution in nature. BAs are involved in numerous vital processes, such as enterohepatic circulation, recognition and transport by various proteins, and their role as “clients” of the farnesoid X receptor, suggesting that they could be used as carriers, transporters, or Trojan horses to deliver a drug to its target. Pioneers of this approach include Ehrlich, Ho, and Kramer, who conceived of “magic bullet” concepts and designed what are now known as conjugated BAs or drug–BA complexes. This review focuses on articles that apply these concepts to the broad and complex field of cancer research. Most of the reviewed studies follow a common trajectory encompassing the design and synthesis of BA conjugates, the in vitro evaluation of their anticancer activity in various cell lines, and their subsequent in vivo assessment. More than 250 compounds have been taken into consideration.
CRISPR-based diagnostics integrated with nucleic acid pre-amplification have demonstrated profound potential for single-molecule detection. However, the pervasive risk of aerosol contamination during amplification significantly hinders their translation to point-of-care testing (POCT). Although amplification-free CRISPR diagnostics promise a streamlined “sample-to-answer” workflow, their development remains in the nascent stages due to the sluggish cleavage kinetics of natural Cas enzymes and the diffusion limitations of trace targets in homogeneous systems. This review systematically summarizes recent core technological advancements, including molecular engineering of CRISPR/Cas systems, novel signal transduction enhancement mechanisms, and digital detection methodologies based on spatial confinement effects. Furthermore, addressing the “matrix effect” that often compromises analytical sensitivity in clinical scenarios, we highlight advanced pre-treatment strategies for complex biological samples. Finally, we propose that the future of POCT relies on the synergy of multiplexed detection, AI-assisted analysis, and microfluidic integration to ultimately bridge the gap between laboratory innovation and clinical application.
Aging is a complex biological process characterized by progressive loss of cellular homeostasis, impaired regenerative capacity, and accumulation of senescent cells that collectively predispose tissues to disease. Traditional two-dimensional culture systems and animal models have provided valuable insights but fail to fully recapitulate the spatial organization, cellular heterogeneity, and microenvironmental cues of aging human tissues. Organoid technology—three-dimensional self-organizing structures derived from adult stem cells or pluripotent stem cells has emerged as a transformative platform to model aging in vitro. These mini-tissues retain the architecture, signaling dynamics, and lineage hierarchy of native organs, making them powerful systems to interrogate age-associated cellular phenotypes, DNA damage responses, and senescence programs. This review discusses how organoid models are advancing our understanding of aging biology across multiple organ systems, from the intestines and liver to the brain and lung. We highlighted key molecular pathways driving cellular senescence within organoids—including p16INK4a/p21CIP1 signaling, SASP activation, mitochondrial dysfunction, and epigenetic drift—and how these can be targeted to restore tissue homeostasis. We further discussed how organoids derived from aged tissues, induced pluripotent stem cells, and engineered oncogene systems reveal new therapeutic opportunities to modulate senescence in age-related disorders, cancer, and regenerative medicine. Finally, we discussed emerging integrative tools such as organoid co-cultures, single-cell omics, and senolytics drug screening that are expanding the potential of organoids as translational platforms for anti-aging and disease intervention.
Background/Objectives: This pilot study investigates the feasibility of using patient-derived microtumors (PDMs) to assess chemotherapy response in epithelial ovarian cancer. Methods: Fresh tissue from 10 patients was used to develop PDMs, which were then tested against carboplatin/paclitaxel, carboplatin/docetaxel, and carboplatin/pegylated liposomal doxorubicin (PLD). Of the 10 PDMs, 3 were obtained from primary debulking surgery (PDS), and 7 were obtained at the time of interval debulking surgery following neoadjuvant chemotherapy. Results: When looking at PDMs derived from tissue collected at the time of PDS, we found that 100% of PDMs demonstrated a full response to carboplatin/PLD, while 30% showed a full response to all regimens, all of which were derived from high-grade serous carcinoma during PDS. The remaining PDMs showed moderate responses to carbo/taxol and carbo/doce. Conclusions: This study suggests that PDMs can be used to assess the efficacy of chemotherapy regimens, as a hypothesis-generating step toward future predictive validation.
Zinc homeostasis is fundamental to metabolic health, orchestrated by the coordinated actions of two major zinc transporter families: ZIP (Zrt- and Irt-like proteins) and ZnT (zinc transporters). ZIP transporters facilitate zinc influx into the cytosol from the extracellular space or from the lumen of intracellular organelles, whereas ZnT transporters control zinc efflux from the cytosol to the extracellular space or facilitate its sequestration into intracellular vesicles and organelles, concurrently harboring the meticulous intracellular zinc homeostasis. This equilibrium is essential for all critical functions like cellular response, metabolic control, and immune pathway alteration. Disruption of this homeostasis is a driver of different pathological alterations like metabolic inflammation, a chronic low-grade inflammatory state underlying obesity; type 2 diabetes; and nonalcoholic fatty liver disease. Recent studies revealed that ZIP and ZnT transporters dynamically regulate metabolic and inflammatory cues, with their tissue-specific expression varying by tissue and acclimating to different physiological and pathological conditions. Recent advanced research in molecular and genetic understanding has helped to deepen our knowledge of the interplay of activity between ZIP and ZnT transporters and their crosstalk in metabolic tissues, underscoring the potential therapeutic prospect for restoring zinc balance and ameliorating metabolic inflammation. This review provides a comprehensive overview that covers the function, regulation, and interactive crosstalk of ZIP and ZnT zinc transporters in metabolic tissues and their pathological conditions.
The indole scaffold represents a privileged structural motif in medicinal chemistry, celebrated for its remarkable chemical versatility, biological ubiquity, and clinical relevance. This review provides a comprehensive analysis of the recent research on the indole nucleus, emphasizing its physicochemical properties, reactivity patterns, and capacity to interact with a wide array of biological targets. Found in key endogenous compounds such as serotonin and melatonin, indole serves as a cornerstone in neurochemical signaling, circadian regulation, and chrono-metabolic homeostasis. Beyond its physiological roles, synthetic indole derivatives have shown extensive therapeutic potential across diverse domains, including oncology, infectious diseases, neurodegenerative disorders, immunomodulation, and metabolic syndromes. The review explores structure–activity relationships (SAR), pharmacokinetics, and the molecular mechanisms by which indole-based compounds exert their tremendous effects, that are ranging from enzyme inhibition to receptor modulation. Special focus is given to current clinical applications and emerging strategies for enhancing drug specificity, bioavailability, and safety through indolic frameworks. Additionally, we highlight the translational potential of indole-containing molecules in personalized medicine, underscoring opportunities for future drug discovery. By integrating insights from medicinal chemistry, biochemistry, pharmacology, and clinical science, this review affirms the indole ring’s enduring value as a central scaffold in therapeutic innovation.
Head and neck cancer (HNC) patients frequently experience alterations in the oral environment following radiotherapy, including xerostomia and impaired mucosal integrity, which may favour fungal overgrowth. This study aimed to characterise oral Candida colonisation in radiotherapy-treated HNC patients and compare it with that of healthy individuals. Unstimulated saliva samples from 61 HNC patients and 100 controls were cultured on chromogenic agar, and isolates were identified using API 20C AUX or MALDI-TOF. Salivary flow was measured to quantify xerostomia. A representative subset of isolates (10 per group) underwent antifungal susceptibility testing by disk diffusion according to CLSI/EUCAST criteria. Candida colonisation was significantly higher in HNC patients than in controls (64.6% vs. 20%, p < 0.001), with greater species diversity and increased detection of non-albicans yeasts, including C. tropicalis, C. parapsilosis, C. glabrata, and C. krusei. All HNC patients exhibited reduced salivary flow. Azole resistance was more frequent among HNC isolates (26%) than among controls (10%), whereas all isolates remained susceptible to amphotericin B and nystatin. These findings indicate that radiotherapy-associated xerostomia substantially alters the oral mycobiota and underscore the importance of routine species-level identification and antifungal susceptibility testing in HNC patients to guide clinical decision-making.
