Search Results (56)

Background and Objectives: Prolonged contact between oral mucosa and dental amalgam restorations may influence local epithelial homeostasis, but the remodeling profile of clinically non-dysplastic mucosa exposed to long-standing amalgam remains insufficiently characterized. This study aimed to evaluate histopathological changes and CK19, Ki67, and p53 expression in the oral mucosa adjacent to long-term amalgam restorations. Materials and Methods: A retrospective observational analysis was performed on 108 oral mucosal specimens, including 78 samples in direct contact with amalgam restorations and 30 non-exposed controls. Exposed cases were grouped according to contact duration: 5–10 years, 11–20 years, and ≥21 years. Histopathological parameters and immunohistochemical expression of CK19, Ki67, and p53 were semi-quantitatively assessed, and an exploratory Integrated Epithelial Remodeling Score was calculated. Results: Longer amalgam exposure was significantly associated with increased inflammatory infiltrate, basal hyperplasia, acanthosis, fibrosis, suprabasal CK19 redistribution, and higher Ki67 labeling indices. The Integrated Epithelial Remodeling Score differed significantly among exposure groups, with higher values in intermediate- and long-duration exposure categories. p53 expression showed statistically detectable but heterogeneous variation. No epithelial dysplasia was observed. Conclusions: Long-term contact with dental amalgam restorations was associated with a coordinated, non-dysplastic remodeling phenotype of the oral mucosa. Given the age imbalance across exposure duration groups, these findings should be interpreted as exposure-associated patterns rather than evidence of a direct causal effect. Because no comparison group exposed to other restorative materials was included, material-specificity for dental amalgam cannot be inferred. In architecturally preserved mucosa, suprabasal CK19 expression may reflect adaptive epithelial plasticity rather than preneoplastic transformation. Full article

Retromer is an evolutionarily conserved protein complex first identified in budding yeast. It was originally described for its essential role in endosome-to-Golgi retrieval of lysosomal hydrolase receptors. Retromer is now known to mediate trafficking of many endosomal cargoes. The mammalian retromer is constituted by a core heterotrimer encoded by the vacuolar protein sorting (VPS) gene products VPS26, VPS35, and VPS29. To mediate cargo recognition and endosomal sorting into various pathways, this trimer can cooperate with phosphoinositide-binding sorting nexin family members. Defective retromer functioning has been associated with alterations in cellular homeostasis, leading to disease. To gain insights into how it may mediate these broad processes, a proteomic strategy in polarized Madin-Darby canine kidney cells was devised to identify retromer-interacting proteins. Subsequent validation of one of the candidates, i.e., cytokeratin 19, led to the unexpected finding that retromer localizes to the pericentriolar region in dividing cells and subsequently translocates to the midbody during cytokinesis. Retromer was found interacting with CK19, and its antisense depletion led to delocalization from CK19. Subcellular fractionation and live cell monitoring of depleted cells provided evidence of a role by retromer in post-metaphase progression and in epithelial cell migration, thereby connecting retromer with key processes of cellular dynamics. Full article

Background/Objectives: The Vesical Imaging-Reporting and Data System (VI-RADS) provides high diagnostic accuracy for muscle-invasive bladder cancer; however, its prognostic value remains limited. We propose serum cytokeratin 19 fragment (CYFRA 21-1) and tumor contact length (TCL) as complementary prognostic factors. We aimed to construct a composite risk score integrating VI-RADS, CYFRA 21-1, and TCL for prognostic stratification. Methods: We retrospectively analyzed data from 101 patients with bladder cancer (BC) who underwent transurethral resection of bladder tumor (TURBT), magnetic resonance imaging, and postoperative serum CYFRA 21-1 measurement. For each factor, cut-off values were determined using receiver operating characteristic (ROC) analysis; meeting each threshold contributed one point (score range, 0–3). Overall survival (OS) was assessed using Kaplan–Meier and Cox regression analyses. Results: ROC analysis identified cut-offs of VI-RADS ≥ 3 (area under the curve [AUC] 0.779), TCL ≥ 40 mm (AUC 0.817), and CYFRA 21-1 ≥ 2.1 ng/mL (AUC 0.875). Based on these, patients were stratified into low- (0–1, n = 81), intermediate- (2, n = 12), and high-risk (3, n = 8) groups with 3-year OS rates of 95.1%, 75.0%, and 25.0%, respectively (p < 0.001). In univariate Cox regression, all factors significantly predicted poor OS: VI-RADS ≥ 3 (hazard ratio [HR], 6.51; p = 0.015), TCL ≥ 40 mm (HR, 8.36; p < 0.001), and CYFRA 21-1 ≥ 2.1 ng/mL (HR, 14.02; p < 0.001). In multivariate analysis, only CYFRA 21-1 remained independently significant (HR, 11.80; p < 0.001). Conclusions: A composite risk score combining VI-RADS, TCL, and CYFRA 21-1 effectively stratified patients with BC into distinct groups using minimally invasive, peri-TURBT assessments. Prospective multicenter validation is warranted. Full article

Salivary gland carcinomas (SGCs) are aggressive malignancies with limited treatment options, primarily due to the complexity of the tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs) remodel the extracellular matrix (ECM), enhance cancer cell stemness, and drive drug resistance. This study introduces a decellularized CAF-derived spheroid system as a biomimetic platform to study tumor–stromal interactions in SGC. Multicellular spheroids were generated by co-culturing Medical Research Council cell strain 5 (MRC-5) fibroblasts (fetal lung-derived) with A253 salivary gland cancer cells, producing distinct spatial architecture, with fibroblasts at the core and cancer cells at the periphery. Compared with A253-only spheroids, A253/MRC-5 spheroids exhibited enhanced proliferation and elevated expression of stemness markers (aldehyde dehydrogenase 1 [ALDH1], CD133, cytokeratin 19 [CK19]). MRC-5 spheroids displayed robust ECM and growth factor expression that persisted after decellularization. Decellularized spheroids retained biological activity, enabling A253 cells to develop invasive phenotypes, metabolic reprogramming, and stemness-associated signatures. Transcriptomic analysis revealed a transition from proliferative pathways to stress-adaptive survival programs, mirroring in vivo tumor behavior. Moreover, A253 cells cultured with decellularized fibroblast spheroids exhibited altered cisplatin sensitivity, highlighting the critical role of stromal ECM in therapeutic response. In conclusion, this study establishes decellularized CAF spheroids as a simplified yet biologically relevant TME-mimetic platform. By recapitulating tumor–stromal crosstalk without live co-culture, this system provides a powerful tool for mechanistic studies of salivary gland cancer, preclinical drug screening, and development of stroma-targeted therapies. Full article

Background: Radon, a naturally occurring radioactive gas, is increasingly recognized as a major risk factor for lung cancer (LC), especially among non-smokers. The objective of this study was to identify serum biomarkers for the early detection of LC in individuals at high risk due to prolonged residential radon exposure in Chiang Mai, Thailand, and to assess whether the use of single or combined biomarkers improves the sensitivity and specificity of detection. Methods: A total of 15 LC patients and 30 healthy controls (HC) were enrolled. The HC group was further stratified into two subgroups: low radon (LR, n = 15) and high radon (HR, n = 15) exposure. All participants were non-smokers or former smokers. Serum levels of cytokeratin 19 fragment (CYFRA 21-1), carcinoembryonic antigen (CEA), interleukin-6 (IL-6), interleukin-8 (IL-8), transforming growth factor-alpha (TGF-alpha), and indoleamine 2,3-dioxygenase-1 (IDO-1) were measured using the Milliplex^® Kit on a Luminex^® Multiplexing Instrument (MAGPIX^® System). Results: Serum CEA, IL-6 and IL-8 levels were significantly higher in LC patients compared to the HC group (p < 0.05). Among analyzed biomarkers, only IL-8 was significantly elevated in LC patients compared to the HR group (p = 0.04). Notably, CYFRA 21-1 was the only biomarker that significantly differed between LR and HR groups (p = 0.004). The diagnostic potential of these biomarkers was evaluated using receiver operating characteristic (ROC) analysis. Individually, IL-6 showed the highest discriminative ability for differentiating LC patients from both HC and HR groups, with high specificity but moderate sensitivity. Combining IL-6 and IL-8 improved specificity and increased the area under the ROC curve (AUC), though it did not enhance sensitivity for distinguishing LC from HC. For distinguishing LC from HR individuals, IL-6 and CYFRA 21-1 exhibited strong diagnostic performance. Their combination significantly improved diagnostic accuracy, yielding the highest AUC, sensitivity, and specificity. In contrast, CEA, IL-8, TGF-alpha, and IDO-1 demonstrated limited diagnostic utility. Conclusions: Based on the available literature, this is the first study to evaluate the combined use of IL-6 and CYFRA 21-1 as potential biomarkers for LC screening in individuals with high residential radon exposure. Our findings highlight their utility, particularly in combination, for improving diagnostic accuracy in this high-risk population. Full article

The mechanisms involved in the regeneration of biliary epithelial cells (BECs) after liver injury remain unclear. In this study, we employed KRT19Cre^ERT/LSL-tdTomato (KT) mice and KT/TFF1KO mice to clarify the regeneration and cell fate of BECs via lineage tracing. Tamoxifen (TAM) was administered to the mice to label cytokeratin 19 (CK19)-positive BECs. The mice were subsequently fed a choline-deficient, ethionine-supplemented (CDE) diet for four weeks, after which the mouse livers were analyzed. Whereas the proportion of tdTomato⁺ cells in CK19-positive BECs decreased in the KT mice, it remained high in the KT/TFF1KO mice. Then, we analyzed hepatic progenitor cells (HPCs), the possible source of BECs. Although tdTomato-labeled HPCs were rarely found in the pretreatment mice, they were frequently found in the KT/TFF1KO mice after the CDE diet, suggesting the dedifferentiation of tdTomato-labeled BECs to HPCs. These results indicate not only that the loss of TFF1 accelerates the dedifferentiation of BECs into HPCs but also that HPCs are the source of BECs in TFF1KO mice. In addition, tdTomato-labeled HNF4α-positive hepatocytes were frequently found in the KT/TFF1KO mice, revealing the transdifferentiation of BECs to hepatocytes. The role of TFF1 as an inducer of biliary differentiation might be useful in the treatment of patients with hepatic or biliary dysfunction. Full article

The Vesical Imaging Reporting and Data System (VI-RADS) is used to detect muscle-invasive bladder cancer, with emerging prognostic implications. Integrating imaging parameters with molecular biomarkers may improve risk stratification in bladder cancer. This study evaluated whether combining VI-RADS scores with serum cytokeratin fragment 19 (CYFRA 21-1) levels—a clinically relevant biomarker for bladder cancer—could improve overall survival (OS) prediction. We retrospectively analyzed 134 patients who underwent transurethral resection of bladder tumors, magnetic resonance imaging, and postoperative serum CYFRA 21-1 measurements. In total, 15 cancer-specific deaths were observed during follow-up. Receiver operating characteristic curve analysis identified optimal prognostic cut-off values: VI-RADS score ≥ 4 and CYFRA 21-1 level ≥ 1.8 ng/mL. The 1-, 2-, and 3-year OS in patients with both high VI-RADS scores and CYFRA 21-1 levels were 42.9%, 16.7%, and 8.3%, respectively, significantly lower than those in other groups (p < 0.001, 0.002, and 0.003, respectively). Multivariate Cox proportional hazards analysis demonstrated that such patients had the poorest OS (hazard ratio: 7.51; p = 0.002). This suggests that combining VI-RADS and CYFRA 21-1 improves prognostic accuracy in bladder cancer, demonstrating potential clinical utility by informing individualized treatment strategies; however, limitations include the retrospective study design and absence of external validation. Full article

Background/Objectives: Lymph node metastases (LNM) undetected by standard hematoxylin and eosin (H&E) have been associated with unfavorable prognosis in colorectal cancer. The One-Step Nucleic Acid Amplification (OSNA) assay has demonstrated superior sensitivity in detecting LNM compared to H&E. We aimed to assess the performance of OSNA in detecting LNM, as well as its prognostic value in rectal cancer (RC) patients. Methods: Lymph nodes (LNs) of patients from 15 centers were analyzed by both H&E and OSNA. The total tumor load (TTL) was defined as the sum of cytokeratin 19 mRNA copies/µL in all LNs from a surgical specimen, using a threshold of 250 copies/μL for OSNA positivity. Cox proportional hazard regression was used to assess the effect of TTL ≥ 250 or 6000 copies/μL on cancer-specific survival (CSS) and recurrence-free survival (RFS), with Firth’s method applied to account for low event rate. Results: A total of 97 RC patients were included. Of these, 84 patients were eligible for survival analysis. The sensitivity and specificity of OSNA, compared to H&E, were 91.7% and 84.7%, respectively. TTL ≥ 6000 versus <6000 copies/μL was related to worse CSS and RFS. When dividing TTL into three groups: ≤250, 250–6000, and >6000 copies/μL, only TTL ≥ 6000 copies/μL was significantly associated with worse CSS and RFS. Conclusions: The OSNA assay is highly sensitive for detecting LNM in RC patients. A TTL of ≥6000 copies/μL could identify a subset of RC patients with worse CSS and RFS who might benefit from adjuvant treatment or intensive surveillance. Full article

Objective: This study aimed to evaluate the reliability and clinical utility of intraoperative sentinel lymph node (SN) metastasis diagnosis using the one-step nucleic acid amplification (OSNA) assay in early-stage cervical cancer by comparing its accuracy with conventional histopathological examination. Methods: A retrospective analysis was conducted on 163 patients who underwent SN biopsy at Kagoshima University Hospital between April 2014 and December 2024. This study included 50 and 113 patients in the OSNA assay and histopathological diagnosis groups, respectively. The OSNA assay quantified cytokeratin 19 (CK19) mRNA levels to determine SN metastasis. The surgical outcomes, SN metastasis detection rates, and non-SN metastasis status were compared between the two diagnostic methods. Results: The SN metastasis detection rate was significantly higher in the OSNA group (12%) than in the pathology group (3%) (p < 0.05). The OSNA assay identified only micrometastases (+) among the positive cases, whereas histopathological diagnosis detected both macrometastases and micrometastases. No non-SN metastases were observed in any of the SN-positive cases, and no significant differences were observed in the recurrence rates between the two groups. Conclusions: The OSNA assay demonstrated a higher SN metastasis detection rate than conventional pathology and demonstrated superior sensitivity in identifying micrometastases. These findings suggest that intraoperative OSNA-based SN assessment in cervical cancer could improve staging accuracy and potentially reduce the need for systematic lymphadenectomy. However, further prospective studies are warranted to confirm these findings and establish clinical guidelines. Full article

Breast cancer is one of the most common cancers diagnosed in both countries with high and low levels of socio-academic development. Routine, regular screening tests being introduced in an increasing number of countries make it possible to detect breast cancer at an early stage of development, as a result of which the trend in the incidence of metastatic breast cancer has been decreasing in recent years. The latest guidelines for the treatment of this tumor do not recommend axillary dissection, which limits the need for rapid assessment of the nodes during surgery. Regardless of the progression of the disease, lymph node biopsy and their analysis is one of the most common diagnostic methods for detecting metastases. Systems using one-step amplification of nucleic acids have been present in the diagnosis of breast cancer for nearly 20 years. The one-step nucleic acid amplification (OSNA) test semi-quantitatively detects the number of cytokeratin 19 mRNA copies, a well-known tumor marker, which can be used to infer the presence of metastases in non-sentinel lymph nodes (SLN). Aim: OSNA is a widely used molecular method for SLN, intra-, or postoperative analysis. Its high accuracy has been proved over the years in clinical use. In this review, we checked current state of this technology and compared it to its competitors in the field of breast cancer diagnosis in the era of Axillary Lymph Nodes Dissection (ALND) importance decrease with intention to foresee its further potential use. Objectives: To evaluate OSNA current place in breast cancer diagnosis and treatment we compared OSNA to other lymph node assessing methods. We based our review on original articles and metanalyses published in the last decade. The research was conducted with PubMed, Science Direct, Google Scholar, and NCBI databases. The collected data allowed us to assess the accuracy of OSNA, its cost effectiveness, and its application in other cancers. Results: Regardless of the progression of the disease, a lymph node biopsy and its analysis constitutes one of the most common diagnostic methods for detecting metastases. The OSNA method is characterized by high sensitivity and specificity, and its predictive value has been confirmed by many studies over the years. While its cost effectiveness is still a matter of discussion, this method has been tested more thoroughly than other new lymph nodes assessing technologies. Conclusions: Despite the emergence of competing methods, this test is still widely used as a routine intraoperative examination of lymph nodes. Research carried out in recent years has proved its effectiveness in the diagnosis of other cancers, in the research field, and as a provider of additional data for prognosis improvement. Full article

Pelvic lymph node dissection (PLND) is the most accurate procedure for lymph node (LN) staging in prostate cancer (PCa) patients. LN sectioning and hematoxylin and eosin (H&E) staining of at least one slice remains the gold standard for LN evaluation, potentially leading to misdetection of small metastatic focus. Entire LN analysis is possible with One-Step Nucleic Acid Amplification (OSNA) by detecting cytokeratin 19 (CK19) mRNA as a surrogate for LN invasion. This study aimed to compare postoperative performance of OSNA pooling with conventional H&E staining for pathological LN detection in PCa patients. POPCORN was an observational, prospective, and multicenter study of patients with PCa who underwent PLND. Dissected LNs were analyzed by both methods. This study included 2503 LNs from 131 patients, showing no statistically significant differences in pathological LN detection. Concordance between methods was high (93.9%), as were specificity (96.6%) and negative predictive value (96.6%) of OSNA pooling. The measure of agreement (Cohen’s Kappa [κ]) was 0.70. Only eight (6.1%) discordances were observed, including four misdetections from each method. Results showed a high concordance between OSNA pooling and H&E staining, suggesting that OSNA pooling may be a good alternative to H&E staining to detect LN metastases in PCa patients. Full article

A novel approach to developing lateral flow assays (LFAs) for the detection of CYFRA 21-1 (cytokeratin 19 fragment, a molecular biomarker for epithelial-origin cancers) is proposed. Magnetic bioconjugates (MBCs) were employed in combination with advanced optical and magnetic tools to optimize assay conditions. The approach integrates such techniques as label-free spectral-phase interferometry, colorimetric detection, and ultrasensitive magnetometry using the magnetic particle quantification (MPQ) technique. For the first time in LFA applications, the MPQ-based and colorimetry-based detection methods were compared side by side, and superior analytical performance was demonstrated. The limit of detection (LOD) of 0.9 pg/mL was achieved using MPQ, and 2.9 pg/mL with optical detection. This study has demonstrated that MPQ provides elimination of signal saturation, higher sensitivity (slope of the calibration curve), and a 19-fold wider dynamic range of detected signals. Both optical and magnetic detection results are comparable to the best laboratory-based tests with the added benefits of a 20-min assay duration and the LFA format convenience. The assay effectiveness was validated in human serum and artificial saliva, and high recovery rates were observed. The proposed approach offers rapid and reliable detection of molecular biomarkers and holds significant potential for point-of-care diagnostics, particularly in resource-limited settings. Full article

Non-small cell lung cancer (NSCLC) is the predominant form of lung cancer and poses a significant public health challenge. Early detection is crucial for improving patient outcomes, with serum biomarkers such as carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCCAg), and cytokeratin fragment 19 (CYFRA 21-1) playing a critical role in early screening and pathological classification of NSCLC. However, due to being mainly based on corresponding antibody binding reactions, existing detection technologies for these serum biomarkers have shortcomings such as complex operations, high false positive rates, and high costs. This study aimed to develop new methods for detecting CEA, SCCAg, and CYFRA 21-1 to assist in the diagnosis of NSCLC. Affinity peptides of CEA, SCCAg, and CYFRA 21-1, respectively, were screened by phage display technology, and the peptides’ binding affinities were determined by enzyme-linked immunosorbent assay and biolayer interferometry. Peptides with high affinity were then integrated as binding domains into biosensors by fusing them with circularly permuted fluorescent proteins (cpFPs) through genetic coding. The resulting biosensors, C4 biosensor for CEA, S1 biosensor for SCCAg, and Y3 biosensor for CYFRA 21-1, demonstrated robust sensitivity and specificity even at concentrations as low as 1 ng/mL for their respective tumor markers. When applied to clinical samples and recalibrated for the upper limit of normal concentrations, the biosensors exhibited enhanced sensitivity and specificity for NSCLC diagnosis. This study introduced innovative biosensors for the detection of CEA, SCCAg, and CYFRA 21-1, providing a highly sensitive, specific, rapid, and cost-effective diagnostic alternative that could significantly improve NSCLC screening rates. Full article

The early detection of tumors is one of the key factors in increasing overall survival in cancer patients. A wide range of cancers still do not have a system of early diagnosis; therefore, the development of new non-invasive tools in this line is essential. Accordingly, the objective of our work was to develop a non-invasive screening method for the early detection of various carcinomas in plasma using a panel that combines two markers using RT-qPCR. A retrospective case-control study was conducted to develop a cancer screening test based on the detection of stromal and epithelial biomarkers (COL1A2 and KRT19) in plasma. The expression of biomarkers was evaluated using multiplex quantitative PCR applied to 47 cases with non-metastatic tumors and 13 control participants. For both biomarkers, a cut-off value was stablished using Youden’s J index through ROC curve analysis and areas under the curve (AUC) were calculated. The plasma mRNA expression level of both biomarkers was significantly higher in diseased versus healthy patients. Moreover, ROC curve analysis showed an AUC value of 0.897 for the combined model. This model also resulted in a cutoff value of 0.664, as well as a sensitivity of 83% and a specificity of 84.6%. These results suggest that the plasma expression levels of COL1A2 and KRT19 could a have potential role in detecting various types of cancer at the early stages. The combined analysis of both stromal and epithelial biomarkers would provide a non-invasive screening method that would allow us to differentiate patients with an active neoplastic process. Full article

A solitary bone plasmacytoma is a rare tumor. Intrahepatic cholangiocarcinoma is the second most common primary liver cancer after hepatocellular carcinoma. We present the case of a 48-year-old female patient who consulted for recent back pain, with a final diagnosis of T10 solitary plasmacytoma and synchronous intrahepatic cholangiocarcinoma. Imaging suggested cholangiocarcinoma with bone metastasis. The patient underwent neurosurgical management with laminectomy, arthrodesis, and arthrectomy, with biopsies revealing monotypic kappa plasmacytic proliferation. Liver biopsies revealed an adenocarcinoma with expression of cytokeratin 19, cytokeratin 7, N-cadherin, and high expression of carbonic anydrase IX. The plasmacytoma was treated with external radiotherapy. The cholangiocarcinoma was treated with selective internal radiation therapy and concomitant systemic treatment with combinations of cisplatin and durvalumab, with capecitabine during radiotherapy, switched for gemcitabine after completion of irradiation. One year after initial management, imaging revealed a partial metabolic response of the intrahepatic cholangiocarcinoma, and a complete metabolic response of the plasmacytoma. This case illustrates the importance of not ignoring two primary tumors and the management of two concomitant treatments exploiting potential therapeutic synergies and limiting expected toxicities. Full article