Identification of Proteomic Biomarkers in Proliferative Verrucous Leukoplakia through Liquid Chromatography With Tandem Mass Spectrometry.

Proliferative verrucous leukoplakia (PVL) is an oral potentially malignant disorder associated with high risk of malignant transformation. Currently, there is no treatment available, and restrictive follow-up of patients is crucial for a better prognosis. Oral leukoplakia (OL) shares some clinical and microscopic features with PVL but exhibits different clinical manifestations and a lower rate of malignant transformation. This study aimed to investigate the proteomic profile of PVL in tissue and saliva samples to identify potential diagnostic biomarkers with therapeutic implications. Tissue and saliva samples obtained from patients with PVL were compared with those from patients with oral OL and controls. Label-free liquid chromatography with tandem mass spectrometry was employed, followed by qualitative and quantitative analyses, to identify differentially expressed proteins. Potential biomarkers were identified and further validated using immunohistochemistry. Staining intensity scan analyses were performed on tissue samples from patients with PVL, patients with OL, and controls from Brazil, Spain, and Finland. The study revealed differences in the immune system, cell cycle, DNA regulation, apoptosis pathways, and the whole proteome of PVL samples. In addition, liquid chromatography with tandem mass spectrometry analyses showed that calreticulin (CALR), receptor of activated protein C kinase 1 (RACK1), and 14-3-3 Tau-protein (YWHAQ) were highly expressed in PVL samples. Immunohistochemistry validation confirmed increased CARL expression in PVL compared with OL. Conversely, RACK1 and YWHA were highly expressed in oral potentially malignant disorder compared to the control group. Furthermore, significant differences in CALR and RACK1 expression were observed in the OL group when comparing samples with and without oral epithelial dysplasia, unlike the PVL. This research provides insights into the molecular mechanisms underlying these conditions and highlights potential targets for future diagnostic and therapeutic approaches.

Nodular fasciitis on the zygomatic region: immunohistochemical analysis and literature review / Fascitis nodular en la región cigomática: análisis inmunohistoquímico y revisión de la literatura

Background: Nodular Fasciitis (NF) is characterized as a benign, fast-growing lesion with proliferation of fibroblasts and myofibroblasts. The use of immunohistochemistry is important for the diagnostic definition and if its findings are not clear, the differential diagnosis will be challenging, even more when the clinical findings do not correspond with the histopathological characteristics. Objective:Here, we reported a case of dermal Nodular Fasciitis affecting zygomatic region of a 64 years old male who complained of swelling in the right side of the face for 3 months, which appeared after an ox-horn trauma. Literature review: We reviewed the literature for all Nodular Fasciitis cases in the zygomatic region. Furthermore, we discussed the relationship of trauma as an etiological factor, main differential diagnoses and immunohistochemical markers for Nodular Fasciitis. Case report: Incisional biopsy was done which revealed benign neoplasm of mesenchymal origin characterized by the fusocellular proliferation. Immunohistochemistry revealed positivity for VIM and SMA, being negative for S-100, CKs, CD34, and p53. The Ki-67 index was low. Due to the clinical, histopathological and immunohistochemical findings, the diagnosis of dermal NF was established. Conclusion: This case consists of Nodular Fasciitis, which must be microscopically differentiated from dermatofibroma, solitary fibrous tumor, low-grade myofibroblastic sarcoma and atypical fibroxanthoma. Immunohistochemistry should always be performed to elucidate the nature of tumor cells and thus contribute to the correct diagnosis and treatment. Nodular Fasciitis appears to be uncommon in the zygomatic region.

Introdução: A Fasciíte Nodular (FN) é caracterizada como uma lesão benigna, de crescimento rápido, com proliferação de fibroblastos e miofibroblastos. O uso da imunoistoquímica é importante para a definição diagnóstica e se seus achados não forem claros, odiagnóstico diferencial será desafiador, ainda mais quando os achados clínicos não corresponderem às características histopatológicas. Objetivo: Relatar um caso de Fasciíte Nodular dérmica acometendo a região zigomática de um homem de 64 anos que se queixava de inchaço no lado direito da face há 3 meses, que surgiu após trauma ocasionado por chifre de boi. Revisão da literatura: A literatura foi revisada para todos os casos de Fasciíte Nodular na região zigomática. Além disso, discutiu-se a relação do trauma como fator etiológico, principais diagnósticos diferenciais e marcadores imunoistoquímicos para Fasciíte Nodular. Relato de caso: Foi realizada biópsia incisional que revelou neoplasia benigna de origem mesenquimal caracterizada pela proliferação fusocelular. A imunoistoquímica revelou positividade para VIM e AML, sendo negativa para S-100, CKs, CD34 e p53. O índice Ki-67 foi baixo. Devido aos achados clínicos, histopatológicos e imunoistoquímicos, foi estabelecido o diagnóstico de Fasciíte Nodular dérmica. Conclusão:Este caso consiste em Fasciíte Nodular, que deve ser diferenciada microscopicamente de dermatofibroma, tumor fibroso solitário, sarcoma miofibroblástico de baixo grau e fibroxantoma atípico. A imunoistoquímica deve sempre ser realizada para elucidar a natureza das células tumorais e assim contribuir para o correto diagnóstico e tratamento. A Fasciíte Nodular parece ser incomum na região zigomática

Intraoral atypical lentiginous melanocytic lesion in a pediatric patient.

We present a rare case of intraoral atypical lentiginous melanocytic lesion affecting a pediatric patient, in which the diagnosis of lentiginous junctional melanocytic nevus with cytologic atypia was favored. The main differential diagnosis is lentiginous melanoma, which is a slowly progressing lesion, affecting mainly older adults, and microscopically presenting lentiginous growth pattern of moderately atypical melanocytes, with focal nesting and pagetoid spread. It is strongly recommended that melanocytic lesions showing features of atypical lentiginous growth pattern should be treated with wide excision; however, the impact of these guidelines on pediatric patients needs to be better defined with the report of further cases.

Post-traumatic lip lesion mimicking rhabdomyomatous mesenchymal hamartoma in a pediatric patient.

We report a 7-year-old boy who presented with a nodule on the upper lip. A previous clinical history of mechanical trauma in the lesional area had been noted. After surgical excision, microscopy revealed fibrocollagenous fascicles associated with neurovascular bundles and skeletal striated muscle fibers in diffuse subepithelial distribution, suggesting rhabdomyomatous mesenchymal hamartoma. However, strict clinicopathological correlation favored a healing process with trapped striated skeletal muscle tissue. After three years of follow-up, an improvement in the aesthetic appearance of the upper lip was observed. To the best of our knowledge, a case of pseudo-rhabdomyomatous mesenchymal hamartoma has not been reported to date.

Activin A triggers angiogenesis via regulation of VEGFA and its overexpression is associated with poor prognosis of oral squamous cell carcinoma.

Poor prognosis associated with the dysregulated expression of activin A in a number of malignancies has been related to with numerous aspects of tumorigenesis, including angiogenesis. The present study investigated the prognostic significance of activin A immunoexpression in blood vessels and cancer cells in a number of oral squamous cell carcinoma (OSCC) cases and applied in vitro strategies to determine the impact of activin A on angiogenesis. In a cohort of 95 patients with OSCC, immunoexpression of activin A in both blood vessels and tumor cells was quantified and the association with clinicopathological parameters and survival was analyzed. Effects of activin A on the tube formation, proliferation and migration of human umbilical vein endothelial cells (HUVECs) were evaluated in gain­of­function (treatment with recombinant activin A) or loss­of­function [treatment with activin A­antagonist follistatin or by stable transfection with short hairpin RNA (shRNA) targeting activin A] conditions. Conditioned medium from an OSCC cell line with shRNA­mediated depletion of activin A was also tested. The profile of pro­ and anti­angiogenic factors regulated by activin A was assessed with a human angiogenesis quantitative PCR (qPCR) array. Vascular endothelial growth factor A (VEGFA) and its major isoforms were evaluated by reverse transcription­qPCR and ELISA. Activin A expression in blood vessels demonstrated an independent prognostic value in the multivariate analysis with a hazard ratio of 2.47 [95% confidence interval (CI), 1.30­4.71; P=0.006) for disease­specific survival and 2.09 (95% CI, 1.07­4.08l: P=0.03) for disease­free survival. Activin A significantly increased tubular formation of HUVECs concomitantly with an increase in proliferation. This effect was validated by reduced proliferation and tubular formation of HUVECs following inhibition of activin A by follistatin or shRNA, as well as by treatment of HUVECs with conditioned medium from activin A­depleted OSCC cells. Activin A­knockdown increased the migration of HUVECs. In addition, activin A stimulated the phosphorylation of SMAD2/3 and the expression and production of total VEGFA, significantly enhancing the expression of its pro­angiogenic isoform 121. The present findings suggest that activin A is a predictor of the prognosis of patients with OSCC, and provide evidence that activin A, in an autocrine and paracrine manner, may contribute to OSCC angiogenesis through differential expression of the isoform 121 of VEGFA.