Search Results (163)

Background/Objectives: Growing evidence indicates that melatonin contributes to kidney development and function, while disruptions of fetal circadian signaling have been linked to congenital anomalies of the kidney and urinary tract (CAKUT). This study aimed to characterize the developmental and spatial expression patterns of melatonin receptors MTNR1A and MTNR1B in normal human fetal kidneys and in CAKUT phenotypes. Methods: This study analyzed 40 human fetal kidney specimens, including healthy controls and CAKUT cases (horseshoe kidneys, duplex kidneys, and dysplastic kidneys), obtained from spontaneous abortions and pregnancy terminations. Samples were classified into developmental phases Ph2–Ph4 according to established morphological criteria. Immunofluorescence staining was used to visualize MTNR1A and MTNR1B expression. Quantitative analysis was performed using ImageJ, measuring the fluorescence area percentage. Statistical comparisons were conducted using a two-way ANOVA. Results: In control kidneys, MTNR1A expression was predominantly observed in glomeruli and interstitial cells and showed a descending trend across developmental stages, whereas MTNR1B was localized to glomeruli and strongly to the apical membranes of tubules, particularly distal tubules, without substantial developmental variation. CAKUT phenotypes exhibited higher expression of both receptors compared to controls. Significant phase-dependent differences in MTNR1A expression were observed in horseshoe, duplex, and dysplastic kidneys. MTNR1B expression decreased across developmental stages in dysplastic kidneys and differed significantly between Ph3 and Ph4 in duplex kidneys. At Ph3, duplex kidneys showed the highest MTNR1B expression. Conclusions: Altered developmental expression patterns of MTNR1A and MTNR1B in CAKUT suggest an association between melatonin signaling and abnormal human kidney development. Full article

Cigarette smoke contains a high concentration of carcinogenic substances to which smokers are regularly exposed. Passive smoking is seriously harmful to the health of non-smoking humans and animals. Domestic cats are particularly vulnerable because of their constant grooming activity, which can promote oral ingestion of smoke-derived residues. Cotinine, a nicotine metabolite, is a reliable biomarker for tobacco exposure. Considering these observations, our study aimed to (1) characterize cytological alterations in oral mucosal epithelial cells by conventional morphology and automated digital cytomorphometry; (2) quantify urinary cotinine concentration and investigate its possible correlation with oral epithelial cytological alterations. To this aim, oral smears were collected from 30 cats divided into two groups (20 exposed; 10 non-exposed). Smears were stained with May–Grünwald–Giemsa and Papanicolaou to assess inflammation and dysplasia; digital cytomorphometric analysis was used to quantify the nucleus-to-cytoplasm (N/C) ratio. Urinary cotinine was measured by ELISA. Our results showed that exposed cats had significantly higher urinary cotinine levels and higher N/C ratios (p < 0.01) than non-exposed controls, along with mild-to-severe inflammation and dysplastic-like epithelial alterations. These findings support urinary cotinine as a valid biomarker of household tobacco smoke exposure in domestic cats and suggest that such exposure may be correlated with early cytological and cytomorphometric changes in the oral mucosa. Further studies are needed to better investigate the relationship between exposure duration and cytological, cytomorphometric, and molecular alterations. Full article

Background/Objectives: Oral squamous cell carcinoma (OSCC) is a major cause of human cancer. The enzyme, nicotinamide N-methyltransferase (NNMT), is overexpressed in a variety of human cancers, including OSCC. Our objective was to target NNMT with novel inhibitors and determine their anti-cancer efficacy while shedding light on their possible mechanism of action. Methods: We identified two small molecule inhibitors of NNMT (AG-670 and AO-022) based on a pharmacophore of the in silico nicotinamide binding site. These inhibitors were investigated for (i) potency to inhibit the activity of the isolated NNMT enzyme (EC₅₀ values), (ii) cytotoxicity (IC₅₀ values) against the human OSCC cell line, SCC-4, and (iii) ability to affect cellular energy metabolism, as measured by oxygen consumption, in SCC-4 cells (plus dysplastic oral keratinocytes (DOK) cells and breast cancer MCF-7 cells). Immunoblotting was used to determine whether NNMT was expressed in the aforementioned cells. Results: NNMT is expressed in SCC-4 and DOK cells (and primary human oral keratinocytes) but not MCF 7 cells. The NNMT inhibitors inhibit isolated NNMT enzyme activity and were cytotoxic to SCC-4 cells (EC₅₀ and IC₅₀ values in the micromolar range). Sublethal doses of the inhibitors were demonstrated to inhibit in situ mitochondrial oxygen consumption in SCC-4 and DOK cells but not in MCF-7 cells. It was demonstrated that the NNMT inhibitors do not directly inhibit mitochondrial electron transport chain activity. Thus, we deduce that the NNMT inhibitors affect mitochondrial activity indirectly via NNMT. Conclusions: It is concluded that NNMT is a potential drug target for oral cancer. Full article

Background/Objectives: The early detection of oral squamous cell carcinoma (OSCC), especially when in the presence of oral potentially malignant disorders (OPMDs), may be challenging and would assist in improving poor OSCC survival rates reported in the literature. We conducted a systematic review to evaluate the utility of adjunctive aids that could assist during clinical examination of the oral cavity to identify suspicious mucosal lesions. Methods: Three databases (CENTRAL, PubMed/MEDLINE, Embase) were screened, limiting results from 2015 to November 2025. Inclusion criteria were: articles written in English; investigating the diagnostic accuracy of diagnostic visual aids compared to conventional oral examination under white light in the assessment of oral mucosal lesions. Extracted data were analysed narratively. Studies not reporting diagnostic accuracy using biopsy results as the gold standard were excluded. Results: The search produced 137 articles; after removing duplicates, 105 were screened through inclusion/exclusion criteria, leading to 17 papers included in the review. Eight articles investigated diagnostic accuracy of narrow band imaging (NBI), seven visually enhanced lesion scopes (VELscopes), one Glasses for Oral Cancer Curing Light Exposed Screening (GOCCLES), one ViziLite chemiluminescence system, and two toluidine blue (TB). Conclusions: High study heterogeneity and lack of randomized clinical trials limit the conclusions of this review. In this context, among the investigated visual aids for expert use, NBI (sensitivity 85–100%, specificity 75–98%) emerges as the most promising tool (VELscope sensitivity 76–87.1%, specificity 21.4–90%; GOCCLES 66%, 48%; ViziLite 77.3%, 27.8%, TB 56.8–91%, 65.3–68%), due to its ability to highlight sub epithelial vascular abnormalities, considered as early indicators of dysplastic or neoplastic progression even. None of the investigated visual aids seem suited for screening purposes/use by the general dentist. Full article

Congenital valve defects account for a substantial proportion of cardiovascular malformations, yet the molecular mechanisms orchestrating cardiac valve development remain incompletely elucidated. While Bone morphogenetic protein (BMP) signaling is essential for valvulogenesis, the specific contributions of individual BMP ligands, particularly the teleost-specific bmp16, have not been characterized. Using the CRISPR/Cas9 system, we generated a bmp16 null knockout and delineated critical roles of this ligand in valvular morphogenesis. bmp16 knockout embryos display a significant reduction in Sox9-positive valvular cells and exhibit severely dysplastic arterial valves, characterized by increased interleaflet distance, thickened leaflets, and shortened leaflet lengths. These morphological abnormalities correlate with impaired valve function, culminating in progressive blood regurgitation, ventricular dilation, and pericardial edema. Mechanistically, loss of bmp16 or pharmacological inhibition of BMP signaling significantly downregulates notch1b expression in developing valves, while pharmacological activation of Notch signaling rescues the regurgitation phenotype in bmp16 mutants. Collectively, our findings establish bmp16 as a novel regulator of valve development and uncover a functional BMP-Notch signaling axis required for vertebrate valvulogenesis, providing new insights into the molecular mechanisms that govern cardiac valve formation and pathogenesis. Full article

Background: Oral squamous cell carcinoma often develops from oral potentially malignant disorders and is frequently diagnosed at an advanced stage. Conventional oral examination is limited by moderate sensitivity, observer variability, and poor discrimination between benign and dysplastic lesions. Adjunctive light-based screening technologies have been introduced, but their diagnostic value remains uncertain. Methods: This systematic review followed PRISMA 2020 guidelines and was registered in PROSPERO. MEDLINE (PubMed), Embase, Scopus, and the Cochrane Database were searched through December 2025. Studies assessing adjunctive light-based screening technologies for detecting oral potentially malignant disorders or squamous cell carcinoma were included. Histopathology served as the reference standard. Diagnostic accuracy outcomes were extracted, and risk of bias was assessed using Cochrane-based criteria. Results: Eleven studies were included. Autofluorescence imaging showed consistently high sensitivity but low and variable specificity. Chemiluminescence demonstrated similar or lower sensitivity with poor specificity. False-positive results were frequent, particularly in inflammatory or benign lesions. Marked heterogeneity across studies limited quantitative synthesis. Conclusions: Adjunctive light-based technologies can increase detection sensitivity when used with conventional oral examination but lack sufficient specificity for standalone use. Histopathological confirmation remains mandatory. Standardized, multicenter diagnostic accuracy studies are needed to clarify their clinical role. Full article

Background: White spot lesions (WSLs) are characterized by enamel demineralization. Minimally invasive treatments using infiltrating resins, such as the commercially available Icon^®, are recommended. The need for such treatments justifies ongoing research into developing materials that can address existing limitations regarding strength, durability, and biocompatibility. Objectives: This study aimed to synthesize and characterize four novel nanobiomaterials by evaluating their physicochemical properties and biocompatibility compared to the commercial material Icon^®. Materials and methods: The recipes for the experimental nanobiomaterials NB3, NB6, NB3F, and NB6F contain varying proportions of TEGDMA, UDMA, HEMA, Bis-GMA, and HAF-BaF2 glass. Mechanical and physicochemical characteristics were evaluated, such as flexural strength, measured using the three-point test; water absorption and solubility; fluoride release; polymerization conversion; and residual monomers, assessed using High-Performance Liquid Chromatography (HPLC). In vitro cell viability was assessed via colorimetry using human dysplastic oral keratinocytes (DOKs). Results: NB6 and NB6F demonstrated the greatest polymerization potential. NB3 exhibited the lowest water absorption and solubility due to its hydrophobic nature. Additionally, the inclusion of UDMA enhanced the strength and elasticity of NB3 when compared to NB6. Among the samples with fluoride additives (NB3F and NB6F), the highest fluoride release on day 7 occurred with the material lacking UDMA. In contrast, the NB3F sample containing UDMA released the least amount of fluoride on the same day. In quantitative terms, NB3 and NB6F exhibited the lowest levels of residual monomers, whereas NB6 showed the highest levels. Both NB3 and NB6 were significantly better tolerated by the cells, showing higher cell viability compared to the commercial material Icon^®. Conclusions: The materials’ mechanical and physicochemical properties varied with component proportions, enabling identification of a suitable formulation for targeted clinical applications. Biocompatibility tests showed that the experimental NB3 and NB6 were better tolerated than Icon^®. Furthermore, the incorporation of filler particles improved the mechanical strength of the experimental nanobiomaterials. Full article

Background/Objectives: Oral potentially malignant disorders (OPMDs), including oral leukoplakia (OLK), oral lichen planus (OLP), and actinic cheilitis (AC), have varying risks of progression to oral squamous cell carcinoma (OSCC). Biomarkers such as aldehyde dehydrogenase 1 (ALDH1) and mammary serine protease inhibitor (Maspin) have shown potential for diagnostic and prognostic use in oral cancer. The present study aimed to evaluate the immunoexpression of aldehyde dehydrogenase 1, a cancer stem cell marker associated with aggressiveness, and the mammary serine protease inhibitor, a potential tumor suppressor, in OPMD and OSCC tissues. Methods: A retrospective analysis was performed on 145 biopsy specimens collected from January 2015 to January 2023, including normal epithelium, OPMDs (OLK, OLP, AC), and OSCC. ALDH1 and Maspin expression levels were evaluated using immunohistochemistry, considering both the percentage of positive cells and staining intensity. Statistical analyses were carried out using the Statistical Package for the Social Sciences (SPSS, version 29.0; IBM Corp., Chicago, IL, USA). Results: Normal oral epithelium showed no expression of ALDH1, whereas 40.6% of OPMDs and 44.4% of OSCC samples exhibited high cytoplasmic ALDH1 expression. Nuclear ALDH1 expression was elevated in 29.7% of OPMDs and 38.9% of OSCCs (p < 0.001). Nuclear Maspin expression was high in 95.2% of normal tissues, in 67.2% of OPMDs and in 55.6% of OSCCs (p < 0.001). Maspin showed strong nuclear and cytoplasmic expression in normal tissue, but its expression decreased in OPMDs and OSCCs, with statistically significant reductions in both compartments (p < 0.001). Conclusions: The results indicate that ALDH1 upregulation and Maspin downregulation are hallmark events in oral carcinogenesis. Their combined evaluation provides a powerful tool for assessing dysplastic severity and malignant transformation risk in OPMDs. Future studies on larger cohorts are needed to confirm the prognostic utility of this dual-marker model. Full article

Background: Oral squamous cell carcinoma (OSCC) is a highly aggressive neoplasm characterized by grim survival outcomes, despite significant therapeutic advances. Mortality rates (up to 70%) have remained unaltered for decades, predominantly due to profound diagnostic delays. These derive from the asymptomatic nature of the early stages of oral carcinogenesis and the emergence of dysplastic areas in previously benign lesions, acting as the bridge to malignant transformation. Hence, the establishment of reliable salivary biomarkers is crucial for non-invasive OSCC detection, even from the premalignant stage of dysplasia. Based on our previous bioinformatic research identifying stage-specific miRNAs throughout OSCC progression, which yielded miR-34a-5p as the most significant, we aimed to experimentally investigate its role in oral oncogenesis and explore its stage-reflecting biomarker potential for liquid biopsy. Methods: The expression of miR-34a was evaluated using quantitative real-time PCR in saliva samples from 9 patients with oral premalignant dysplastic lesions, 10 patients with OSCC and 10 healthy controls. The diagnostic accuracy of miR-34a expression profiles was assessed using ROC-curve analyses. Results: The expression of salivary miR-34a differed significantly across the studied groups, demonstrating a steep decrease in the presence of epithelial premalignant dysplasia, significant upregulation in OSCC and intermediate levels in normal oral mucosa (p < 0.001). The ROC results indicate strong diagnostic performance for the detection of oral dysplasia (AUC = 0.93; p < 0.001), OSCC (AUC = 0.77; p = 0.01) and excellent accuracy for the discrimination between premalignant and OSCC lesions (AUC = 0.98; p < 0.001). Conclusions: Our findings reveal a state-dependent dysregulation of miR-34a in oral carcinogenesis, suggesting its complex role as a pathogenetic agent that allows for malignant transformation through its diminished expression, and as a secondary reactive mechanism attempting to suppress tumor development. Salivary miR-34a holds great, stage-specific diagnostic potential, thereby reflecting the health state of oral mucosa in real time. Full article

Head and neck cancer represents a heterogeneous group of malignancies. Oral squamous cell carcinoma (OSCC) is the most prevalent form of head and neck cancer, with a rising incidence in recent years. Risk factors for developing OSCC include exposure to carcinogens, such as alcohol and tobacco products, that can lead to molecular alterations in the oral mucosa and progression from premalignant lesions to invasive phenotypes. Despite the relative curative potential of localized OSCC, the overall prognosis of OSCC has not significantly improved for decades due to a frequently delayed diagnosis and limited targeted treatment options. There remains a need to better characterize the molecular biomarkers of OSCC progression, especially in dysplastic mucosal lesions, before their malignant transformation. In this review, we discuss several molecular biomarkers highly implicated in OSCC tumorigenesis that have demonstrated correlation with clinicopathological parameters and clinical outcomes. These biomarkers are typically involved in vital pathways of carcinogenesis, including cell cycle control, growth factor signaling, and stress responses. They include ubiquitous cancer biomarkers such as p53 and PTEN, as well as those more specific to OSCC, such as DJ-1 and Cornulin. Collectively, we envision that a diverse panel of these biomarkers can provide the greatest clinical benefit in enhancing early detection and prognostic accuracy, while some individual biomarkers may also serve as therapeutic targets for personalized approaches to head and neck cancers. Full article

Background and Clinical Significance: Mastocytosis and mast cell activation syndrome (MCAS) include conditions in which patients manifest signs, symptoms, and laboratory findings consistent with mast cell activation and can only be diagnosed in the presence of specific criteria. Mutations of ZRSR2, a gene involved in RNA splicing, are not closely associated with mast cell disorders, but rather with myelodysplastic syndromes development. Case Presentation: We report a case of a 37-year-old man who was referred to our institution for anaphylaxis after a bee sting and elevated serum tryptase levels (17.8 ng/mL in the first sample and 19.2 ng/mL in the second sample). Complete blood count was unremarkable. Bone marrow biopsy showed signs of dysplasia and some CD25+ mast cells. ASO-qPCR and targeted myeloid NGS analysis did not detect the KIT p.D816V mutation, but rather showed the presence of a pathogenetic variant of the ZRSR2 gene (p.S447_R448del) with a variant allele frequency of 7.4%. Mastocytosis could not be diagnosed based on the established diagnostic criteria. The patient’s symptoms were not recurrent and tryptase release was not event-related; therefore, a diagnosis of MCAS could not be made either. Taken together, these findings led to the diagnosis of clonal hematopoiesis of indeterminate potential (CHIP). A watch and wait strategy consisting of clinical evaluations, blood tests, and cardiovascular risk assessment was initiated. Conclusions: This case report highlights the importance of combining clinical and laboratory findings, hematopathology, and molecular analyses to establish the most probable diagnosis in challenging cases. It also underscores the possible relevance of identifying predisposing conditions, such as CHIP, in order to guide counseling and follow-up strategy. Full article

Background: During development, reelin sets the pace of neocortical neurogenesis, enabling newborn neurons to migrate. However, whether—and, if so, how—reelin signaling affects the adult neurogenic niches remains uncertain. Methods: In the present study, we use both loss- and gain-of-function genetic approaches, along with in vivo and ex vivo assays, to investigate this question. Results: We show that reelin signaling, resulting in Dab1 phosphorylation, occurs in the ependymal-subependymal zone (EZ/SEZ) of the lateral ventricles, where, along with its associated rostral migratory stream (RMS), the highest density of functional ApoER2 accumulates. Mice deficient in Reelin, ApoER2, or Dab1 exhibit enlarged ventricles and a dysplastic RMS. Moreover, while the conditional ablation of Dab1 in neural progenitor cells (NPCs) enlarges the ventricles and impairs neuroblast clearance from the SEZ, the transgenic misexpression of Reelin in NPCs of Reelin-deficient mice normalizes the ventricular lumen and the density of ependymal cilia, thereby ameliorating neuroblast migration. Consistently, intraventricular infusion of reelin reroutes neuroblasts. Conclusions: These results demonstrate that reelin signaling persists, sustaining the germinal niche of the lateral ventricles and influencing neuroblast migration in the adult brain. Full article

Background/Objectives: Cervical Pap smears are routinely used to detect cellular abnormalities as a cervical cancer screening tool and to assess the presence of HPV for risk stratification of the disease. Here, we aimed to extend the applications of this sampling procedure by combining it with multicolor flow cytometry to characterize cell populations across cervical cancer disease stages. Methods: Cervical Pap smears from 30 patients with various disease stages ranging from normal to intraepithelial neoplasia up to treated cancers were analyzed as biofluids using multicolor flow cytometry. Individual samples were evaluated, and statistical analyses were performed over all sample stages. Cancer cell lines (CaSki, SiHa, HeLa, A549, U2OS) were examined as tumor cell controls. Results: Cervical biofluids were subdivided into cell populations according to their scattering properties and the expression of specific biomarkers: EpCAM and cytokeratin 8 for epithelial cells from tumors as well as healthy ectocervical and endocervical regions, and CD45 for immune cells. Discrimination of tumor cells was facilitated with cancer cell lines. Statistical analysis revealed that the composition of cell populations differs among disease stages, whereas treated cancer samples were consistently associated with a reduction in squamous epithelial cells and an increase in immune cells compared to normal samples. Conclusions: Herein, we identified the major cell populations in cervical biofluid samples and demonstrated that this method can detect changes in the cellular composition across different disease stages. This approach could be further exploited in cancer research and potentially serve as a companion diagnostic tool in tumor development, progression and during treatment. Full article

Background/Objectives: PRAME (Preferentially Expressed Antigen in Melanoma) is a promising immunohistochemical marker for distinguishing melanoma from benign melanocytic lesions, though optimal thresholds remain uncertain. This study evaluated PRAME expression in melanocytic lesions and compared diagnostic accuracy using two thresholds. Methods: We retrospectively assessed PRAME expression in 145 melanocytic lesions diagnosed between 2016 and 2021 at Istanbul Training and Research Hospital: 52 melanomas, 27 dysplastic nevi, 23 Spitz nevi, 15 compound nevi, 23 blue nevi, and 5 congenital nevi. Immunohistochemical staining (PRAME EP461, Cell Marque) was scored semi-quantitatively based on nuclear positivity: 0 (negative), 1 (1–24%), 2 (25–49%), 3 (50–74%), and 4 (≥75%). Diagnostic accuracy was evaluated at 50% and 75% thresholds. Results: PRAME expression at both thresholds was significantly higher in melanomas than nevi (p < 0.05). Sensitivity and specificity were 92.3% and 96.8% at 50%, and 82.7% and 98.9% at 75%. Lowering the threshold to 50% improved sensitivity with minimal specificity loss, particularly differentiating melanoma from dysplastic, compound, and blue nevi. Occasional positivity was observed in Spitz and dysplastic nevi; one melanoma was PRAME-negative. Conclusions: PRAME is an effective marker for melanoma diagnosis. A 50% threshold optimizes sensitivity while preserving specificity; however, histopathological evaluation remains the gold standard, and PRAME should be used only as an adjunct to avoid potential overdiagnosis, particularly in borderline lesions. Full article

Venous malformations (VMs) are rare, non-involuting, slow-flow, congenital anomalies of vascular morphogenesis, presenting as dilated venous channels with reduced perivascular cell coverage. The treatment may be conservative or surgical, including laser therapy. The management of small superficial VMs typically involves surgical excision. In larger or deeper VMs, the intralesional–endovascular injection of the sclerosing agent bleomycin is the gold standard, as it eliminates the dysplastic venous vessels by inducing fibrosis and therefore promotes regression of the lesion. This review explores the current literature regarding the role of bleomycin in venous malformation management, emphasizing the molecular pathways involved, the efficacy of sclerotherapy with bleomycin and its complications and the associated management challenges. It evaluates the clinical and histological features of venous malformations, alongside diagnostic methodologies and treatment strategies, drawing on the most recent bibliographic data. The literature was systematically reviewed using the PubMed database, offering insights into future research directions and highlighting innovative treatment approaches. Full article