RESUMEN
The uncertainty of true labels in medical images hinders diagnosis owing to the variability across professionals when applying deep learning models. We used deep learning to obtain an optimal convolutional neural network (CNN) by adequately annotating data for oral exfoliative cytology considering labels from multiple oral pathologists. Six whole-slide images were processed using QuPath for segmenting them into tiles. The images were labeled by three oral pathologists, resulting in 14,535 images with the corresponding pathologists' annotations. Data from three pathologists who provided the same diagnosis were labeled as ground truth (GT) and used for testing. We investigated six models trained using the annotations of (1) pathologist A, (2) pathologist B, (3) pathologist C, (4) GT, (5) majority voting, and (6) a probabilistic model. We divided the test by cross-validation per slide dataset and examined the classification performance of the CNN with a ResNet50 baseline. Statistical evaluation was performed repeatedly and independently using every slide 10 times as test data. For the area under the curve, three cases showed the highest values (0.861, 0.955, and 0.991) for the probabilistic model. Regarding accuracy, two cases showed the highest values (0.988 and 0.967). For the models using the pathologists and GT annotations, many slides showed very low accuracy and large variations across tests. Hence, the classifier trained with probabilistic labels provided the optimal CNN for oral exfoliative cytology considering diagnoses from multiple pathologists. These results may lead to trusted medical artificial intelligence solutions that reflect diverse diagnoses of various professionals.
Asunto(s)
Aprendizaje Profundo , Redes Neurales de la Computación , Humanos , Citodiagnóstico/métodos , Procesamiento de Imagen Asistido por Computador/métodos , Neoplasias de la Boca/diagnóstico , Neoplasias de la Boca/patología , PatólogosRESUMEN
BACKGROUND: Squamous cell carcinoma (SCC) of the dorsum of the tongue is extremely rare, and it clinically resembles various benign lesions. Somatic mutations in TP53 and some driver genes were implicated in the development of SCC; however, the somatic genetic characteristics of dorsal tongue SCC remain unknown. With a detailed analysis of gene mutations in dorsal tongue SCC, we aimed to better understand its biology. METHODS: Four cases of SCC initially occurring on the tongue dorsum were evaluated for clinical and histological findings and immunohistochemical expression of p53 and p16. Gene mutations were analyzed using next-generation sequencing with a custom panel of driver genes. RESULTS: We retrospectively investigated 557 cases of tongue SCC, and only four cases of SCC initially occurred on the tongue dorsum. The four patients (cases 1-4) were one woman and three men with a mean age of 53.75 years (range: 15-74 years). Histological analysis revealed well-differentiated SCC. Through molecular analysis, we identified pathogenic somatic mutations, namely, TP53 p.C176F (c.527G > T) in case 3 and TP53 p.R282W (c.844 C > T) in case 4. No pathogenic variants were identified in the PI3K/AKT or RAS/RAF pathways. The p53 immunohistochemical examination revealed a wild-type expression pattern in cases 1-3 and strong expression in case 4. The results of p16 immunostaining were negative in all cases. CONCLUSIONS: We described four previously unreported genetic characteristics of dorsal tongue SCC. Somatic TP53 mutations may contribute to the development of a subset of dorsal tongue SCC; however, more cases with genetic analysis need to be accumulated.
Asunto(s)
Carcinoma de Células Escamosas , Mutación , Neoplasias de la Lengua , Proteína p53 Supresora de Tumor , Adolescente , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/análisis , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patología , Inhibidor p16 de la Quinasa Dependiente de Ciclina/genética , Análisis Mutacional de ADN , Secuenciación de Nucleótidos de Alto Rendimiento , Inmunohistoquímica , Estudios Retrospectivos , Carcinoma de Células Escamosas de Cabeza y Cuello/genética , Carcinoma de Células Escamosas de Cabeza y Cuello/patología , Lengua/patología , Neoplasias de la Lengua/genética , Neoplasias de la Lengua/patología , Proteína p53 Supresora de Tumor/genéticaRESUMEN
Hyperplastic dental follicles (HDFs) represent odontogenic hamartomatous lesions originating from the pericoronal tissues and are often associated with impacted or embedded teeth. These lesions may occasionally feature unique calcifying bodies, known as calcifying whorled nodules (CWNs), characterized by stromal cells arranged in a whorled or spiral fashion. CWNs are typically observed in multiple calcifying hyperplastic dental follicles or regional odontodysplasia. In our study, we examined 40 cases of HDFs, including nine instances with characteristics of CWNs, referred to as calcifying hyperplastic dental follicles (CHDFs), which are infrequently accompanied by odontodysplasia. The median ages of the HDFs and CHDFs were 16 (ranging from 3 to 66) and 15 (ranging from 11 to 50) years, respectively. The lower third molars were the most frequently affected by HDSFs and CHDFs, followed by the upper canines. A histological examination was conducted on all 40 cases, with an immunohistochemical analysis performed on 21 of them. Among the cases with CWN, nine affected a single embedded tooth, with one exception. CWNs exhibited diverse calcifications featuring sparse or entirely deposited psammoma bodies, and some displayed dentinoid formation. Immunohistochemically, the stromal cells of HDFs were frequently positive for CD56 and nestin. By contrast, CWNs were negative for CD56 but positive for nestin as well as hairy and enhancer split 1 (HES1), with a few dentin sialoprotein (DSP)-positive calcified bodies. Our results revealed that hamartomatous CHDFs can impact multiple and single-embedded teeth. CWNs composed of nestin and HES1-positive ectomesenchymal cells demonstrated the potential to differentiate into odontoblasts and contribute to dentin matrix formation under the influence of HES1. This study is the first report documenting odontoblastic differentiation in HDFs. The rare occurrence of HDFs and CHDFs contributes to limited comprehension. To prevent misdiagnosis, a better understanding of these conditions is necessary.
RESUMEN
Sclerosing odontogenic carcinoma (SOC) is an exceedingly rare odontogenic carcinoma known for its locally aggressive yet indolent behavior. There have been no reports of metastasis to distant organs, except a single case involving lymph node metastasis. This report details the case of a 49-year-old female who presented with a well-demarcated radiolucent lesion in the mandible, accompanied by root resorption and tooth displacement. Microscopically, the lesion exhibited a distinctive composition, with two distinct components: cords of epithelium embedded within an abundant collagenous stroma and solid nests of clear polygonal cells surrounded by hyalinized stroma. Notably, the tumor exhibited direct invasion into the submental muscles, accompanied by perineural and vascular invasion, as well as cortical bone loss. Additionally, the clear cells contained diastase-sensitive periodic acid-Schiff-positive granules. Immunohistochemically, the tumor cells displayed positivity for cytokeratin 19 and p63 while testing negative for myoepithelial markers. The Ki-67 index was measured at 23%. Importantly, neitherEWSR1 nor MAML2 rearrangements were detected through fluorescence in situ hybridization (FISH) analysis. Over several years, this patient experienced three instances of local recurrence; notably, four years after the initial surgery, fludeoxyglucose F18-positron emission tomography (18FDG-PET)/CT scans confirmed the presence of pulmonary metastasis. This case presents an unusual histological variation of SOC, marked by vascular invasion, and is notably the first documented case of a fatal outcome in this context.
RESUMEN
BACKGROUND: Odontogenic keratocysts constitute 10%-20% of odontogenic cysts and exhibit a distinctive corrugated parakeratinized lining epithelium. Considering that cornified envelope formation is an important phenomenon during keratinocyte differentiation, this study aimed to clarify the characteristics of cornified envelope formation in odontogenic keratocysts. METHODS: We investigated the cellular distribution of cornified envelope-related proteins (transglutaminases and their substrates), as well as the upstream regulatory protein c-Fos, by immunohistochemical analysis of the lining epithelium of 20 odontogenic keratocysts. We examined the corresponding mRNA levels by quantitative polymerase chain reaction. Ten dentigerous cysts served as control non-keratinized cysts. RESULTS: The distributions of transglutaminase and their substrates except loricrin and small protein-rich protein 1a significantly differed between odontogenic keratocysts and dentigerous cysts. There was no significant difference in c-Fos expression between odontogenic keratocysts and dentigerous cysts. The mRNA levels of transglutaminases and their substrates were significantly higher in odontogenic keratocysts than in dentigerous cysts. However, c-Fos mRNA levels did not significantly differ between groups. CONCLUSION: Surprisingly, the overall appearance of cornified envelope-related proteins of odontogenic keratocysts was consistent with the characteristics of non-keratinized oral mucosa identified in previous studies. These findings indicate that the contribution of cornified envelope-related molecules in odontogenic keratocysts is similar to that in non-keratinized oral epithelium, rather than keratinized oral epithelium, suggesting that odontogenic keratocysts are not genuine keratinized cysts. The upregulation of cornified envelope-related genes in odontogenic epithelium could be an important pathognomonic event during odontogenic keratocyst development.
Asunto(s)
Quiste Dentígero , Quistes Odontogénicos , Humanos , Quiste Dentígero/patología , Quistes Odontogénicos/genética , Quistes Odontogénicos/patología , Epitelio/patología , TransglutaminasasRESUMEN
Glomeruloid hemangioma is a rare variant of hemangioma that is accompanied by polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin abnormalities (POEMS) syndrome and, rarely, by thrombocytopenia, anasarca, fever, reticulin fibrosis, and organomegaly (TAFRO) syndrome. This report presents the case of a 78-year-old male who presented with a hemorrhagic nodule on the tongue without any other systemic diseases. Microscopically, the lesion was a lobular proliferation extending from the lamina propria to muscular tissue. Some intravascular nodules with irregular vascular lumens closely resembled renal glomeruli. Each nodule consisted of plump endothelial and stromal cells that partially showed vacuolated cytoplasm containing eosinophilic and periodic acid-Schiff (PAS)-positive globules. Immunohistochemically, IgG-positive deposition was noted within CD31-positive cells. Many plump stromal cells were positive for CD31, CD146, nestin, and type IV collagen but not α-smooth muscle actin (αSMA). These results reflect the proliferation of immature endothelial cells and pericytes, which might characterize this unique lesion. Microscopically, this case revealed glomeruloid hemangioma without systemic conditions related to POEMS, and composed of an intravascular proliferation of immature endothelial and pericytic stromal cells.
RESUMEN
PURPOSE: This study aims to delve deeper into the hypothesis that normal salivary gland tissue expresses both protein and mRNA of mammaglobin (MGB). METHODS: Formalin-fixed paraffin-embedded samples of submandibular (10), parotid (5), palatal (5) and labial glands (30) salivary glands were immunohistochemically investigated. The labial samples were used to examine the MGB positive ratio (MGB-PR), and localize MGB by double immunofluorescence staining and quantitative mRNA gene expression. Mann-Whitney U and Kruskal Wallis rank-sum test for group comparison, and Spearman's rank correlation coefficient for correlation analysis were used. RESULTS: The distribution of MGB-positive cells was variable throughout samples with significantly higher MGB-PR of acini than ducts (P = 0.00376), and there was no difference when compared based on age (P = 0.0646) and gender (P = 0.245). Besides acinar cells, a number of myoepithelial cells and ductal cells also demonstrated strong MGB reactivity with varying MGB mRNA expression levels in 6 of the 7 samples (with MGB-PR > 20%) tested. CONCLUSION: This novel study shows that unlike aberrant protein expression in some carcinomas, MGB expression in salivary gland neoplasms represents the nature of original cells, giving a better insight into the neoplasms expressing MGB.
Asunto(s)
Glándula Parótida , Glándulas Salivales , Células Epiteliales , Expresión Génica , ARN MensajeroRESUMEN
Cornified envelope formation is crucial for the final differentiation of keratinized epithelium. However, the mechanisms of cornified envelope formation in the oral epithelium remain unclear. The aim of this study was to clarify the differences in the distribution and expression of cornified envelope related proteins and genes between keratinized and non-keratinized oral epithelia. We immunohistochemically investigated the distribution patterns of transglutaminase 1 (TG1), transglutaminase 3 (TG3), and their substrate proteins involucrin (IVL), loricrin (LOR), and small proline rich proteins (SPRs), in 19 keratinized and 14 non-keratinized oral epithelium samples. TG1 and TG3 mRNA levels were investigated in both types of epithelium by real time reverse transcription polymerase chain reaction (RT-PCR) using paraffin-embedded specimens. Data were analyzed to identify factors involved in cornified envelope formation. We demonstrate that 11 localization patterns show statistically significant differences between keratinized and non-keratinized oral epithelia. These factors clearly drove the separation of the two groups during cluster analysis. TG1 mRNA levels in keratinized oral epithelium were significantly higher than those in non-keratinized oral epithelium. In conclusion, the characteristic distribution of transglutaminases and their substrates and the mRNA levels of TG1 can regulate cornified envelope formation in keratinized oral epithelium, together with the contribution of TG3 first reported in this paper.
Asunto(s)
Mucosa Bucal , Transglutaminasas , Diferenciación Celular , Membrana Celular , Epitelio , QueratinocitosRESUMEN
Several investigators have reported that oral membranous and pharyngeal viscous deposits developed in bedridden elderly persons requiring nursing care without oral intake. Therefore, this study aimed to clarify the origin of viscous deposits on the pharyngeal mucosa based on characteristics of salivary and tracheal secretory mucin. The participants were 35 elderly people who required nursing care. All 46 collected specimens, including 30 intraoral and 16 pharyngeal specimens, were stained against specific mucins secreted from the respiratory tract and saliva gland using antibodies anti-MUC2 and anti-MUC7, respectively. Out of 35 participants, the intraoral membranous deposits and deposits on the pharyngeal mucosa developed in 17 (48.6%) and 10 persons (28.6%), respectively. The pharyngeal deposits developed in 58.8% of participants who developed intraoral deposits. All pathological specimens shared microscopic findings of various combinations of eosinophilic lamellar structure and a pale-basophilic amorphous substance. Immunohistochemically, both the 30 oral and the 16 pharyngeal specimens obtained from 17 participants were consistently positive for MUC7 but negative for MUC2. In conclusion, we clarified that the mucoid component of both oral and pharyngeal deposits comprised MUC7 salivary mucin, which revealed that both deposits originated from the oral cavity. This result strongly suggests that oral care is intimately related to oral and pharyngeal conditions.
Asunto(s)
Mucinas , Proteínas y Péptidos Salivales , Anciano , Humanos , Boca , Mucina 2 , Faringe , Glándulas SalivalesRESUMEN
OBJECTIVES: The aim of this study was to clarify by histopathological examination the origin of oral membranous substances deposited on the palate, tongue, buccal mucosa and teeth. BACKGROUND: Several investigators have reported membranous substances deposited in the mouths of bedridden elderly persons requiring nursing care without oral intake. However, the precise nature and origin of the substances are poorly understood. METHODS: Sixty-nine specimens were taken from the oral cavity of bedridden patients, that is, the palate, dorsum of the tongue, the cheek and teeth. Sections were stained with haematoxylin and eosin stain, alcian-blue and periodic acid-Schiff stain (AB-PAS) and antibodies for pankeratin (AE1AE3) and leukocyte common antigen (LCA). RESULTS: All specimens showed a film-like nature coloured from tan to white, accompanied by a mucous substance. Histologically, specimens of all sites had a similar feature of the combination of basophilic amorphous and eosinophilic lamellar features. The basophilic substance was positive for AB-PAS, and PAS-positive glycogen granules were also noted in the lamellar structure. Immunochemistry revealed various degrees of pankeratin positive substance and LCA-positive inflammatory cell infiltration. CONCLUSION: The oral membranous substance was composed of keratin and mucin with inflammation. These results suggest that the deposition of the oral membranous substance is a pathological condition or oral mucositis caused by dry mouth.
Asunto(s)
Personas Encamadas , Mucosa Bucal/patología , Hueso Paladar/patología , Nutrición Parenteral , Lengua/patología , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Japón , Queratinas/análisis , Masculino , Mucosa Bucal/química , Mucinas/análisis , Hueso Paladar/química , Lengua/química , Diente/química , Diente/patología , Xerostomía/patologíaRESUMEN
OBJECTIVE: Oral lichen planus (OLP) characterized by interface mucositis frequently shows hyper-keratinization. To clarify mechanisms of excess keratinization, we investigated key molecules for cornified cell envelope (CE). METHODS: Involucrin (IVL), loricrin (LOR), transglutaminase 1 (TGase 1) and transglutaminase 3 (TGase 3) were immunohistochemically examined in 20 specimens of OLP; five specimens of buccal mucosa served as controls. Subsequently, the data were statistically analyzed. RESULTS: IVL in OLP was localized in the cell membrane, in contrast to its localization in the cytoplasm in controls. No positive reaction indicative of LOR was noted in any specimens. Although the TGase 1 localization in controls was restricted to the upper three-quarters of the membrane, the localization in OLP was in both membrane and in the cytoplasm of full thickness mucosal layers. The TGase 3 localization pattern was dramatically altered from cytoplasmic to membranous in OLP. CONCLUSION: Our data suggest that aberrant TGase 1 and TGase 3 localization and distribution are closely related to hyper-keratinization in OLP. This is the first report of ectopic transglutaminase localization in OLP.
Asunto(s)
Queratinas/metabolismo , Liquen Plano Oral/metabolismo , Transglutaminasas/metabolismo , Membrana Celular/metabolismo , Epitelio/metabolismo , Humanos , Espacio Intracelular/metabolismo , Persona de Mediana Edad , Precursores de Proteínas/metabolismoRESUMEN
In oral lichen planus, extracellular matrix and basal cells are damaged by T-lymphocytes. As a consequence, changes in expression of collagen fibers within the connective tissue and cytoskeletons of the epithelial tissue can be observed. With the goal of examining the characteristic changes undergone by basal cells as a consequence of T-lymphocytes damage in oral lichen planus, we investigated protein expression in the epithelial-connective junction. We selected 20 cases of oral lichen planus and 5 control samples of buccal mucosa. Subsequently, we divided the oral lichen planus cases into thin and thick parts based on the mean values of epithelial thickness from the control samples, and counted the positive rate of collagen IV, keratin 19, desmoglein 1, and Ki-67. Collagen IV immune-reactivity partially disappeared or thickened in oral lichen planus. The keratin 19 positive rate in oral lichen planus cases was significantly lower than in the controls. Desmoglein 1 positive rate of the thick part was significantly higher compared to the thin part of oral lichen planus. Thus, modifications in basal cells with both reduced keratin 19 expression and alterations of desmoglein 1 expression suggest that in oral lichen planus, as a consequence of cell injury or regeneration in the interface area, there is a disappearance of the "true basal cell nature".