Fine needle aspiration cytology (FNAC) is widely used to evaluate various lesions. Rapid on-site evaluation (ROSE) can be incorporated into cytology procedures. Our study aims to determine the efficacy of ROSE on the specimen adequacy of FNAC as compared to non-ROSE cases. We conducted a prospective study that included consecutive FNAC cases from various sites between January 10th, 2025, and April 9th, 2025. All cases performed in our laboratory's FNAC clinic had undergone ROSE using a Diff-Quik stain, whereas all FNAC cases in which the glass slides were received from other institutions for cytological evaluation were not subjected to ROSE. The specimen adequacy of ROSE-performed FNAC cases was compared with that of the non-ROSE group. This study included 350 patients. ROSE-performed cases constituted 30.8% (n = 108), while non-ROSE cases accounted for the remaining 69.2% (n = 242). The thyroid was the most frequently sampled site overall (38.86%, n = 136), followed by lymph nodes (31.14%, n = 109), breast (16.58%, n = 58), salivary gland (9.14%, n = 32), and soft tissue (3.14%, n = 11). Cell block preparations were performed in 22.85% (n = 80) of cases, of which 88.75% (n = 71) belonged to the ROSE group. Among the 286 overall adequate cases (81.7%), ROSE was performed in 106 cases (98.14% specimen adequacy, p < 0.001), while 180 out of 242 cases in the non-ROSE group were adequate (74.38% specimen adequacy, p < 0.001). The current study validates ROSE as a relevant and beneficial method that can be used as an adjunct to FNAC to improve the specimen adequacy.

This study aimed to retrospectively analyze the prevalence and clinical characteristics of TUGSE and explore the potential role of viral agents in these lesions. Fifty-seven TUGSE and 50 non-specific ulcer patients were included. Patient demographic and clinical data were collected. Formalin-fixed paraffin-embedded tissues were screened by PCR for Epstein-Barr virus (EBV), cytomegalovirus (hCMV), human papillomavirus (HPV), herpes simplex virus (HSV), and Helicobacter pylori (H. pylori). The majority of TUGSE patients were in their sixth (29.8%) and seventh (24.6%) decades of life, with a 1:1.6 male-to-female ratio. The tongue is the most common site (47.4%), followed by buccal mucosa (28.1%). The average duration was 13.5 weeks, and almost one-fourth of the lesions were clinically diagnosed as oral squamous cell carcinoma (24.6%). TUGSE showed a significantly higher propensity to harbor macrophage infiltrates and inflammatory involvement with skeletal muscle fibers or salivary gland tissues, compared to non-specific ulcers. EBV was observed in two TUGSE cases (3.5%) and one patient with a non-specific ulcer (2.0%). Other viral agents were undetectable in TUGSE. TUGSE frequently affects the elderly population with a female predilection. The observed low prevalence of viral agents suggests that these pathogens may not play a direct role in TUGSE development.

Background: Oral mucoceles are common benign lesions of the minor salivary glands, frequently involving the lower lip. They arise due to trauma-induced rupture or obstruction of salivary ducts, leading to mucus accumulation within the surrounding connective tissue. Although usually asymptomatic, mucoceles may cause aesthetic concern, functional interference, and recurrence. Aim: To present a clinical case series of lower lip mucoceles managed surgically and to review their pathogenesis, diagnosis, and management. Case Series: Two patients presenting with painless lower lip swellings were clinically diagnosed as mucoceles. Both cases were treated by complete surgical excision. Healing was uneventful, with no recurrence on follow-up. Conclusion: Surgical excision of oral mucoceles along with the associated minor salivary gland tissue remains the most predictable treatment modality, offering excellent prognosis and minimal recurrence.

Innovative molecular diagnostic strategy for salivary gland lesions based on ATR-FTIR spectroscopy coupled with machine learning.

Background/Objectives: The Notch-ADAM17 pathway is a fundamental signaling mechanism where ADAM17, a disintegrin and metalloprotease, cleaves the Notch receptor after the Notch receptor binds to a ligand. Crosstalk between Notch and ADAM17 is often altered in pathological situations. Alterations in Notch2 expression, in particular, appears to be correlated with the onset of various autoimmune diseases. In primary Sjögren's disease (pSjD), an autoimmune disorder characterized by chronic inflammation, the role of ADAM17 has been extensively explored, but a correlation with Notch2 has not yet been evaluated. Methods: To analyze the gene and protein expression of Notch2 in pSjD and a possible correlation with ADAM17 expression and with the patient's inflammatory grade, we employed an integrated co-detection protocol to analyze salivary gland tissue sections by combining in situ hybridization (ISH) with immunohistochemistry (IHC). Results: combined ISH/IHC allows us to demonstrate an increased expression of Notch2 mRNA and protein in pSjD salivary glands (SGs) biopsies, which appears correlated with an increased expression of ADAM17, both in acinar and duct cells and in infiltrating lymphocytes. Notch2/ADAM17 expression is higher in biopsies of pSjD SGs characterized by a high degree of inflammation. Conclusions: this work demonstrates the correlated expression in pSjD SGs of ADAM17, which plays multiple roles in the pathogenesis of SjD, and Notch2, widely considered a key player in various inflammatory mechanisms, offering a starting point for future therapeutic interventions to investigate.

BackgroundSjögren syndrome (SS) is a common autoimmune disease characterized by lymphocytic infiltration. Describing the transcriptional and metabolic features of the disease from a spatial perspective can enhance our understanding of the disease pathogenesis and treatment;MethodsWe collected eight human labial samples, including four labial gland samples from patients with SS and from four healthy controls. We integrated single-cell RNA sequencing, spatial transcriptomics, and spatial metabolomics techniques to generate SS-associated spatial gene expression maps and spatial metabolite profiles at a single-cell resolution. We also analyzed the characteristic metabolic and genetic changes of SS samples and infiltrated CD4+ T cells. Immunohistochemistry was used to detect the infiltration of CD4+ T cells in the salivary glands. In vivo experiments were conducted using NOD/ShiLtj mice to validate the therapeutic effects of targeting PS(36:1) on SS-like symptoms, as assessed by HE staining, salivary flow rate assays, and immunofluorescence experiments.ResultsComprehensive data from spatial multi-omics identified the cell types and distributions within the immune microenvironment of salivary glands in SS. CCL19 was significantly increased in lymphocyte infiltration, while IGHG4 was elevated in glandular area. Linoleic acid metabolism undergoes reprogramming in SS, with alterations in lecithin, linoleic acid, 13(S)-hydroxyoctadecadienoic acid and dihomo-γ-linolenate. Furthermore, PS (36:1) was found to be abnormally enriched in lymphocyte focus, which may be related to the abnormal expression of CD74 and HLA-DRA. Also, CXCL13 corresponded to areas resembling high levels of PS(36:1) in infiltrated CD4+T cells. We further investigated the effect of CD4+ T cells on the reprogramming of glycerophospholipid metabolism in SS, including key regulatory genes and key metabolites. Among them, LYPLA2 and PS(36:1) exhibited the most representative results. In vivo, the PS(36:1) synthesis inhibitor alleviated dry mouth symptoms and reduced lymphocyte infiltration in the salivary gland tissues of NOD/ShiLtj mice, while also suppressing CD4+ T cell accumulation in the infiltrating foci.ConclusionsThe multi-omics analysis conducted in this study enhances our understanding of the key regulatory mechanisms driving the pathogenesis of SS and offers novel insights for its precision therapy. Furthermore, PS(36:1) emerged as a potential therapeutic target for future SS research and treatment.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12967-025-07361-x.

Spermidine enhances Immunoglobulin A secretory capacity in rat salivary glands: An ex vivo study.

Sjögren disease (SjD) causes salivary hypofunction and ocular dryness. To investigate mechanisms of autoimmunity and salivation, we profiled unstimulated whole saliva from SjD patients and demographically matched controls by 4D-DIA proteomics and untargeted LC-MS/MS metabolomics. Unstimulated whole saliva from patients meeting the 2016 ACR-EULAR criteria for SjD and demographically matched controls was profiled by 4D-DIA proteomics and untargeted LC-MS/MS metabolomics. An independent validation cohort (newly diagnosed SjD, n = 24; controls, n = 24) underwent ELISA for salivary ZG16B, a subset provided labial minor salivary gland (LSG) tissue for immunofluorescence (IF). Proteomics identified 2032 differentially expressed proteins (1355 up; 677 down). Gene Ontology indicated downregulation of epithelial differentiation, structural maintenance, and epithelial tube formation, while KEGG highlighted enrichment of protein catabolism, immune regulation, and protein processing pathways. ZG16B was significantly reduced in SjD. Metabolomics detected 702 differential metabolites (307 up; 385 down); upregulated pathways involved protein digestion/absorption and multiple amino-acid metabolisms, whereas downregulated pathways mapped to aerobic respiration, suggesting reduced energy production. In validation, salivary ZG16B levels were markedly lower in newly diagnosed SjD than controls, and IF showed absent or sparse ZG16B-positive acinar cells in SjD LSG. Salivary ZG16B correlated positively with tear and salivary flow rates and inversely with focus score (FS). Integrated proteo-metabolomic analysis reveals concurrent epithelial injury, immune activation, and compromised energy metabolism in SjD salivary glands. ZG16B downregulation is associated with exocrine dysfunction, supporting ZG16B as a promising noninvasive biomarker for SjD. Key Points • Saliva multi-omics reveals epithelial injury, immune activation, and metabolic impairment in SjD. • ZG16B is consistently downregulated in saliva and salivary gland epithelium. • ZG16B shows high diagnostic accuracy and mirrors gland function/histologic damage. • Dysregulated proteasomal and amino-acid pathways link molecular changes to secretory dysfunction.

Microbubble (MB) technology is uniquely suited for integration into microphysiological systems (MPS) for high throughput three-dimensional (3D) tissue culture, drug screening and toxicity testing. MBs are spherical compartments with nanoliter volumes produced in an array format. Here, we present a novel hybrid MB-fluidic MPS that combines 3D tissue culture with controlled fluid flow to improve nutrient delivery, waste removal, and physiological relevance. Computational fluid dynamics (CFD) simulations were used to model velocity and solute diffusion profiles as a function of MB aspect ratio (AR), with validation by fluorescent polystyrene microsphere optical tracking. Simulations reveal pronounced velocity decoupling and shear dampening effects with intra-MB flow velocities over 200-fold lower than the main channel-allowing high channel flow rates for efficient exchange while preserving low-shear microenvironments, optimal for tissue culture. Additionally, tissues spatially compartmentalized in individual MBs are not dislodged under high flow conditions. This allowance for high channel flow rates, decoupled from the MB microenvironment, enables the use of millifluidic devices which are less difficult to manufacture and control than microfluidic devices. Simulations also showed that MBs with AR values between 2 and 3 offered a balance between nutrient transport and retention of cell-secreted factors. In contrast, rectilinear wells exhibited flow splitting and lactate accumulation at AR > 2, highlighting a key advantage of the spherical MB geometry. We fabricated a millifluidic MB device using fused deposition modeling (FDM) 3D printing and a novel molding strategy to create optically clear, leak-free flow channels. Murine salivary gland tissues cultured under flow in this device showed preserved acinar cell marker gene expression and reduced ductal markers, supporting the hypothesis that dynamic flow enhances tissue fidelity. This MB-fluidic platform enables scalable, high-content 3D culture systems suitable for organoid, tumor spheroid, and tissue mimetic applications in drug discovery and toxicology.

Ixodes ricinus ticks (Acari: Ixodidae) are hematophagous ectoparasites and a major European vector of zoonotic diseases affecting global health. Tick salivary and midgut proteins antigen p23 (A0A0K8RKR7) and metalloprotease (A0A0K8RCY8) were previously implicated in the pathophysiology of the tick-borne allergy alpha-Gal syndrome (AGS). This study aimed to functionally characterize these two biomolecules, focusing on their role in I. ricinus tick feeding and reproduction through gene knockdown by RNA interference and midgut transcriptomic analysis. Validation of RNA-seq data was conducted using RT-qPCR analysis on tick midgut and salivary gland tissues. Silencing the expression of p23 and metalloprotease did not result in any significant differences in tick engorgement and egg batch weights compared to the control group. Gene set enrichment analysis following antigen p23 gene knockdown identified significantly upregulated pathways associated with protein production and suppressed routes mostly correlated with ion transport, lipid metabolism, catalytic activity, protein modification, and G-protein activity. Partial knockdown of the metalloprotease led to the upregulation of several biological and functional pathways associated with RNA splicing and significantly suppressed routes connected with detoxification, protein modification, catalytic activity and molecule binding. Antigen p23 appears to play a functional role in tick midgut cell homeostasis, primarily by participating in regulatory and signaling processes essential for cell viability. Metalloprotease is potentially involved in regulating midgut response to oxidative stress, thereby reducing tissue damage and promoting regular cell proliferation, growth and behavior. These results provide insights into tick physiology and bases for further research on tick-host interactions and AGS pathogenesis.

Organoid models have significantly enhanced our understanding of adult stem cell function, however, uncovering regulatory mechanisms governing rare and often quiescent stem cells in glandular organs remains challenging. Here, we employ an integrative multi-omics approach, combining single-cell RNA sequencing, bulk ATAC and RNA sequencing, to profile the cellular populations and signaling pathways characterizing a mouse salivary gland organoid model across different temporal stages and after radiation-induced damage. Our findings identify Sox9- and Itgb1/Cd44-expressing cells as primitive adult stem/progenitor populations with a critical migratory role in tissue repair. Notch signaling is a key driver of self-renewal and migration in response to irradiation. Additionally, scRNA-seq analysis of irradiated salivary gland tissue confirms these findings in an in vivo setting. Extending these findings to murine and patient-derived salivary, mammary and thyroid gland organoids, we reveal the conserved role of Notch signaling in coordinating stem/progenitor cell-mediated regeneration across glandular tissues. These insights position Notch signaling as a central regulator of glandular stem cell-like populations and as a promising therapeutic target for enhancing glandular tissue regeneration following cancer therapies.

Disclosure: C. Rizk: None. G. Sydney: None. P. Gopal: None. S. Omay: None. P. Balasubramanian: None.Background: Ectopic salivary tissue within the sella is rare and mostly asymptomatic. Preoperative diagnosis is often challenging as they mimic pituitary neoplasms and the diagnosis is typically made after resection or biopsy. We present a case of ectopic intrasellar salivary gland tissue confirmed on histopathology in a patient who was presumed to have a meningioma on her initial imaging performed for the evaluation of headache. Case Presentation: A 61-year-old female presented with 4 month history of severe bifrontal headache and bilateral upper extremity weakness. Brain MRI was performed revealing a 2.8 x 2.6 x 1.9 cm mass along the left petroclival region extending into the sella, and into the left cavernous sinus with deviation of the pituitary stalk. The MR imaging characteristics were consistent with meningioma. Endocrine biochemical evaluation was unremarkable. Given the discordance between the patient’s clinical presentation and imaging characteristics and the short duration of her symptoms, endoscopic endonasal biopsy of the sellar lesion was performed for a definitive diagnosis. Histopathology was consistent with benign seromucinous salivary gland tissue with chronic inflammatory infiltrate. To rule out metastatic disease, DOTATATE PET scan was done which showed tracer uptake within the sella and the left cavernous sinus but no other abnormality. The patient was managed conservatively with improvement in her symptoms, and the lesion has remained stable on serial MRI imaging over the last one year. Conclusion: Ectopic salivary gland tissue in the sellar region is a rare finding and is typically asymptomatic. When symptoms do occur, they are often attributed to leakage of glandular secretions leading to localized inflammation. In uncommon instances, larger lesions may cause endocrine disturbances, including hyperprolactinemia or growth hormone deficiency. Definitive diagnosis generally relies on histopathological evaluation, and the overall prognosis is excellent.Presentation: Sunday, July 13, 2025

Benign tumours of the salivary glands most commonly affect the parotid gland. Diagnostic evaluation should include ultrasound-guided fine needle aspiration, with additional imaging as needed. The most frequent tumour types are pleomorphic adenoma and Warthin's tumour. This review finds that treatment typically involves surgical excision, although in selected cases, a watchful waiting approach may be appropriate. There is currently no consensus on the optimal surgical technique for all cases. Complete tumour removal, while preserving adjacent nerves and healthy salivary gland tissue, remains essential.

BackgroundMost head and neck cancer patients treated with ionizing radiation loose salivary gland function. Patients with decreased saliva have trouble eating, speaking and are predisposed to oral infections and tooth decay. Amifostine is the only FDA approved drug to prevent radiation-induced hyposalivation. However, it has intolerable side-effects that limit its use, motivating the discovery of alternative therapeutics.MethodsWe leveraged our salivary gland tissue chip platform for high-content drug discovery that we developed using submandibular gland tissue from female SKH1 hairless mice, backcrossed 6 generations with C57BL/6 J mice. We developed in-chip assays to quantify reduced glutathione and cellular senescence, which are accepted biomarkers of radiation damage. We validated radioprotection using WR-1065, the active form of Amifostine and tested other reported radioprotective drugs including Edaravone, Tempol, N-acetylcysteine, Rapamycin, Ex-Rad, and Palifermin. Next, a Selleck Chemicals library of FDA-approved drugs was screened for radioprotection. Lead hits were tested in mouse models.ResultsWe identify 25 candidate compounds and down-select them using EC50 values and published pharmacologic data. This lead us to test Phenylbutazone (an anti-inflammatory), Enoxacin (a fluoroquinolone antibiotic), and Doripenem (a carbapenem antibiotic) for in vivo radioprotection in mice. Results confirm that these three drugs exhibit radioprotection equivalent to Amifostine but with superior EC50 values, ranging from 140 to 6900-fold lower values.ConclusionsThis body of work demonstrates the development and validation of assays using a tissue chip platform for high-content drug screening and the successful discovery and in vivo validation of candidate radioprotective drugs with non-antioxidant primary modes of action. These results point to possible unknown mechanisms of radioprotection. These drugs can be developed to improve radioprotection efficacy and clinical administration without adverse side-effects.

BackgroundSolid tumors comprise approximately 60% of all pediatric cancers. Relapsed or refractory tumors of the central nervous system (CNS), such as atypical teratoid/rhabdoid tumors (AT/RTs), are the leading cause of death in children with cancer. Claudin 6 (CLDN6)-specific chimeric antigen receptor (CAR) T cells have demonstrated activity in preclinical and clinical studies in various solid adult cancers. However, the suitability of CLDN6 as a target in pediatric tumors and their susceptibility to CAR T-cell therapy has yet to be established. This study aimed to evaluate the suitability of CLDN6 as a target for CAR T-cell therapy of pediatric solid tumors.MethodsImmunohistochemical CLDN6 expression was assessed in fetal normal tissues (n=91), pediatric normal tissues (n=157), and two sets of pediatric tumor tissues (n=527 and n=49) using a combined score that includes the percentage of stained cells with a 4-point intensity scale (0 to 3+). The antitumor activity of CLDN6 RNA-transduced CAR T cells against AT/RT cell lines was assessed with in vitro assays and in immunodeficient NOD-SCID-γc–/– (NSG) mouse models bearing orthotopic xenograft tumors.ResultsMembranous CLDN6 expression, as detected by immunohistochemistry, was widely observed in fetal tissues but was absent in almost all non-malignant pediatric tissues, except for very rare, scattered cells with 1+ to 2+ intensity in kidney, pancreas, pituitary, and salivary gland tissues. Membranous CLDN6 expression was frequently detected in a subset of the pediatric tumor entities, including germ cell tumors (93% of samples with CLDN6-positive cells), nephroblastoma (64%), extracranial malignant rhabdoid tumors (50%), and AT/RTs (39%). In CLDN6-positive samples, CLDN6 was generally expressed with 2+ or 3+ intensity in substantial proportions of the cancer cells. Strong CLDN6 expression was also detected in single samples of hepatoblastoma, Ewing sarcoma/other embryonal tumors, and osteosarcoma.In experimental models, CLDN6-CAR T cells led to antigen-specific killing of endogenously CLDN6-expressing AT/RT cell lines in vitro and exhibited potent and specific antitumor activity in mice bearing orthotopic CLDN6-expressing AT/RT xenograft tumors.ConclusionsThese results support CLDN6 as an oncofetal cell-surface antigen that may be suitable for CAR T-cell targeting in pediatric solid tumors, including those of the CNS.

BackgroundDisappointing outcomes in Sjögren’s disease (SjD) trials underscore the need for reliable, sensitive endpoints. Histological assessment holds promise, but a minimally invasive, repeatable method for salivary gland tissue sampling is lacking.ObjectivesTo evaluate the feasibility, safety and tissue adequacy of ultrasound-guided core needle biopsy (US-guided CNB) of the parotid gland and explore its role for facilitating histology-driven stratification and precision medicine.MethodsIn the Belgian Sjögren’s Syndrome Transition Trial, 66 patients (64 without gland swelling) underwent US-guided CNB. US was evaluated using OMERACT (Outcome Measures in Rheumatology Clinical Trials). and Hocevar scoring. Histopathology included assessment of focus score, B cell predominance (CD20>CD3), follicular dendritic cell networks (CD21), plasma cells (CD138), lymphoepithelial lesions (CK7/CK14) and FcRL4+ B cells. Pain was assessed using a visual analogue scale (VAS) from 0 to 10. Findings were matched with clinical data.ResultsMean VAS pain scores were 2.7 (SD=2.77) during biopsy and 1.9 (SD=2.33) in the 3 days before the follow-up call at day 14. No major complications occurred, and 82% of patients were willing to repeat the procedure. Adequate tissue was retrieved in 62/66 cases. Patients showed histological heterogeneity and were, as proof of concept, stratified into mild, moderate and severe histological involvement. Histological severity correlated with ultrasound scores (p<0.01) and not with traditional outcome measures (European Alliance of Associations for Rheumatology Sjögren’s Syndrome Patient-Reported Index dryness and European Alliance of Associations for Rheumatology Sjögren’s Syndrome Disease Activity Index).ConclusionUS-guided CNB is safe, well-tolerated and yields adequate tissue. Beyond diagnostics, it might facilitate histology-driven patient stratification and advance precision medicine for SjD.

Metformin mitigates inflammation and apoptosis in salivary gland epithelial cells via an AMPK-dependent mechanism in chronic obstructive sialadenitis.

High activity of human cytomegalovirus in patients with Sjögren's disease.

Background: Pleomorphic adenoma and mucoepidermoid carcinoma are the most common benign and malignant salivary gland tumors, respectively. The role of cancer stem cells and autophagy in tumor progression and aggressiveness is crucial. Nucleostemin, a stem cell-enriched nucleolar protein, plays a role in stemness, proliferation, and tumor progression however, its role in salivary gland tumorigenesis remains unexplored until now. Materials and Methods: This study evaluated nucleostemin immunohistochemical expression in 27 samples, including normal salivary gland tissue, pleomorphic adenoma, and mucoepidermoid carcinoma. Nucleostemin immunoexpression was analyzed via Leica Qwin 500 software. Statistical analysis using ANOVA test, Bonferroni post hoc correction test, were performed to assess differences across groups. Results: Nucleostemin nuclear expression was highest in mucoepidermoid carcinoma (27.55 ± 1.91), followed by pleomorphic adenoma (16.89 ± 1.64), and showed the least expression in normal salivary glands (0.47 ± 0.08). Conclusion: This research revealed notable expressions of nucleostemin in mucoepidermoid carcinoma and pleomorphic adenoma suggesting its potential role in tumor biology. Further studies with larger samples and clinicopathological correlation are recommended.

Prospects and challenges of salivary gland tissue engineering in Sjögren's syndrome.