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Resumen ANTECEDENTES: Los leiomiomas son los tumores pélvicos más frecuentes en la mujer; sin embargo, su localización vaginal es excepcional. Suelen ser asintomáticos y encontrarse como un hallazgo clínico. En los últimos 20 años solo se han reportado 85 casos, y solo dos han sido recurrentes. OBJETIVO: Reportar un caso de miomatosis vaginal recurrente en una paciente histerectomizada y revisar la bibliografía al respecto. CASO CLÍNICO: Paciente de 58 años, histerectomizada, con una tumoración vaginal. El reporte histopatológico informó una proliferación fusocelular, debidamente delimitada, dispuesta en haces entrecruzados. Los núcleos eran alargados, monomorfos y de extremos romos. El estroma era escaso y colagénico. No se observaron atipias citonucleares ni necrosis. El estudio inmunohistoquímico de la lesión con actina de anticuerpos antimúsculo liso y desmina se reportó positivo. Se diagnosticó miomatosis vulvovaginal recurrente. Se trató mediante resección quirúrgica. CONCLUSIÓN: Los leiomiomas vulvovaginales son extremadamente raros y la bibliografía al respecto es poca; su recurrencia es verdaderamente excepcional. De ahí la importancia de la publicación de estos casos, que aporta información que pueden tomar en cuenta otros clínicos al momento del diagnóstico.

Abstract BACKGROUND: Leiomyomas are the most frequent pelvic tumors in women; however, their vaginal location is unusual. They are usually asymptomatic and present as a clinical finding. In the last 20 years only 85 cases have been reported, and only two have been recurrent. OBJECTIVE: To report a case of recurrent vaginal myomatosis in a hysterectomized patient and review the literature. CLINICAL CASE: A 58-year-old hysterectomized patient with a vaginal tumor. The histopathologic report reported a fusocellular proliferation, properly delimited, arranged in crisscross bundles. The nuclei were elongated, monomorphous and blunt ended. The stroma was sparse and collagenous. No cytonuclear atypia or necrosis were observed. Immunohistochemical study of the lesion with anti-smooth muscle antibody actin and desmin was reported positive. Recurrent vulvovaginal myomatosis was diagnosed. It was treated by surgical resection. CONCLUSION: Vulvovaginal leiomyomas are extremely rare, and the literature is sparse; their recurrence is truly exceptional. Hence the importance of publishing these cases, providing information to be considered by other clinicians at the time of diagnosis.

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ABSTRACT Objective: The aim of our research is to assess fascin expression in nasal tissues of patients with chronic rhinosinusitis with (CRSwNP) and without (CRSsNP) nasal polyps. Methods: Fascin expression in nasal tissues of 11 CRSwNP patients and 10 CRSsNP patients was immunohistochemically evaluated and compared with control subjects. Results: Fascin was found to be strongly expressed in epithelial cells in polyps in CRSwNP and nasal tissue in CRSsNP. Its strong expression was observed both in lamina propria and nasal epithelial cells in CRSsNP. Fascin overexpression in nasal mucosa in CRSwNP was more pronounced compared with CRSsNP. In addition, proliferating epithelial cells in polyp tissue were weakly immunostained, whereas mature cells expressed much more fascin. Conclusion: CRSwNP and CRSsNP are associated with fascin overexpression, which makes fascin a promising target for therapeutic interventions.

RESUMEN Objetivo: El objetivo de esta investigación fue evaluar la expresión de fascina en los tejidos nasales de pacientes con rinosinusitis crónica con (RSCcPN) y sin pólipos nasales (RSCsPN). Métodos: La expresión de fascina en los tejidos nasales de 11 pacientes con RSCcPN y 10 pacientes con RSCsPN fue analizada por inmunohistoquímica y comparada con los individuos control. Resultados: Fascina fue encontrada por ser fuertemente expresada en células epiteliales de pólipos en la RSCcPN y en tejido nasal en la RSCsPN. Su fuerte expresión fue observada tanto en la lámina propia como en las células epiteliales nasales en la RSCsPN. La sobreexpresión de fascina en la mucosa nasal en la RSCcPN fue más pronunciada en comparación con la RSCsPN. Además, las células epiteliales proliferantes en el tejido del pólipo fueron inmunoteñidas débilmente, mientras las células maduras expresaron mucho más fascina. Conclusión: RSCcPN y RSCsPN están asociadas a la sobreexpresión de fascina, lo que hace la fascina un objetivo prometedor para intervenciones terapéuticas.

RESUMO Objetivo: O objetivo desta pesquisa foi avaliar a expressão de fascina nos tecidos nasais de pacientes com rinossinusite crônica com (RSCcPN) e sem (RSCsPN) pólipos nasais. Métodos: A expressão de fascina nos tecidos nasais de 11 pacientes com RSCcPN e 10 pacientes com RSCsPN foi avaliada imuno-histoquimicamente e comparada com os indivíduos-controle. Resultados: Fascina foi encontrada por ser fortemente expressa em células epiteliais em pólipos na RSCcPN e em tecido nasal na RSCsPN. Sua forte expressão foi observada tanto na lâmina própria quanto nas células epiteliais nasais na RSCsPN. A superexpressão de fascina na mucosa nasal na RSCcPN foi mais pronunciada em comparação com a RSCsPN. Além disso, as células epiteliais em proliferação no tecido do pólipo foram imunocoradas fracamente, enquanto as células maduras expressaram muito mais fascina. Conclusão: RSCcPN e RSCsPN estão associadas à superexpressão de fascina, o que torna a fascina um alvo promissor para intervenções terapêuticas.

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Abstract Objective: To determine the role of the dishevelled binding antagonist of beta catenin 1 (DACT1) in the cytoskeletal arrangement of cardiomyocytes in atrial fibrillation (AF). Methods: The DACT1 expression and its associations with the degree of fibrosis and β-catenin in valvular disease patients were analyzed by immunohistochemistry and Masson's staining. DACT1 was overexpressed in the atrial myocyte cell line (HL-1) and the cardiac cell line (H9C2) by adenoviral vectors. Alterations in the fibrous actin (F-actin) content and organization and the expression of β-catenin were detected by flow cytometry, immunofluorescence, and Western blotting. Additionally, the association of DACT1 with gap junctions connexin 43 (Cx43) was detected by immunohistochemistry, immunofluorescence, and Western blotting. Results: Decreased cytoplasmic DACT1 expression in the myocardium was associated with AF (P=0.037) and a high degree of fibrosis (weak vs. strong, P=0.028; weak vs. very strong, P=0.029). A positive association was observed between DACT1 and β-catenin expression in clinical samples (P=0.028, Spearman's rho=0.408). Furthermore, overexpression of DACT1 in HL-1 and H9C2 cells induced an increase in β-catenin and subsequent partial colocalization of DACT1 and β-catenin. In addition, F-actin content and organization were enhanced. Interestingly, DACT1 was positively correlated with the Cx43 expression in clinical samples (P=0.048, Spearman's rho=0.370) and changed the Cx43 distribution in cardiac cell lines. Conclusion: DACT1 proved to be a novel AF-related gene by regulating Cx43 via cytoskeletal organization induced by β-catenin accumulation in cardiomyocytes. DACT1 could thus serve as a potential therapeutic marker for AF.

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Introduction: Leptin is a hormone secreted by adipocytes that has been associated with the epithelial-mesenchymal transition (EMT). Additionally, leptin promotes the migration and invasion of mammary epithelial cells through the activation of FAK and Src kinases, which are part of a regulatory complex of signaling pathways that promotes the expression of proteins related to the formation of proteolytic structures involved in the invasion and progression of cancer. Recently, overexpression and activation of Hic-5 during the EMT have been shown to induce the formation of actin puncta; these structures are indicative of the formation and functionality of invadopodia, which promote the local degradation of extracellular matrix components and cancer metastasis. Objective: To evaluate the role of FAK and Src kinases in the expression of Hic-5 during the epithelial-mesenchymal transition induced by leptin in MCF10A cells. Materials and methods: We used specific inhibitors of FAK (PF-573228) and Src (PP2) to evaluate Hic-5 expression and subcellular localization by Western blot and immunofluorescence assays and to investigate the formation of actin puncta by epifluorescence in MCF10A cells stimulated with leptin. Results: Leptin induced an increase in Hic-5 expression and the formation of actin puncta. Pretreatment with inhibitors of FAK (PF-573228) and Src (PP2) promoted a decrease in Hic-5 expression and actin puncta formation in the non-tumorigenic mammary epithelial cell line MCF10A. Conclusion: In MCF10A cells, leptin-induced Hic-5 expression and perinuclear localization, as well as the formation of actin puncta through a mechanism dependent on the kinase activity of FAK and Src.

Introducción. La leptina es una hormona secretada por los adipocitos que se ha relacionado con el proceso de la transición de epitelio a mesénquima (Epithelial-Mesenchymal Transition, EMT). Promueve la migración e invasión de las células del epitelio mamario mediante la activación de las cinasas FAK y Src, un complejo regulador de vías de señalización que favorecen la expresión de las proteínas relacionadas con la formación de estructuras proteolíticas implicadas en la invasión y progresión del cáncer. Recientemente, se ha descrito que la sobreexpresión y activación de la proteína Hic-5 durante el mencionado proceso de transición, favorece la formación de los puntos de actina (indicativa de la formación y funcionalidad de los invadopodios), lo cual promueve la degradación local de los componentes de la matriz extracelular y la metástasis del cáncer. Objetivos. Evaluar el papel de las cinasas FAK y Src sobre la expresión y localización subcelular de Hic-5 y la formación de puntos de actina inducida por la leptina en la línea celular MCF10A de epitelio mamario no tumoral. Materiales y métodos. Se utilizaron los inhibidores específicos de la FAK (PF-573228) y la Src (PP2) para evaluar el papel de ambas cinasas en los niveles de expresión y localización subcelular de la proteína Hic-5 mediante Western blot e inmunofluorescencia, así como la formación de puntos de actina mediante la tinción con faloidina-TRITC en células MCF10A estimuladas con leptina. Resultados. La leptina indujo el incremento en la expresión de Hic-5 y la formación de puntos de actina. El tratamiento previo con los inhibidores de las cinasas FAK (PF-573228) y Src (PP2), promovió la disminución en la expresión de Hic-5 y de los puntos de actina en la línea celular MCF10A de epitelio mamario no tumoral. Conclusión. La leptina indujo la expresión y la localización perinuclear de Hic-5 y la formación de puntos de actina mediante un mecanismo dependiente de la actividad de las cinasas FAK y Src en las células MCF10A.

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OBJECTIVE: To determine the role of the dishevelled binding antagonist of beta catenin 1 (DACT1) in the cytoskeletal arrangement of cardiomyocytes in atrial fibrillation (AF). METHODS: The DACT1 expression and its associations with the degree of fibrosis and ß-catenin in valvular disease patients were analyzed by immunohistochemistry and Masson's staining. DACT1 was overexpressed in the atrial myocyte cell line (HL-1) and the cardiac cell line (H9C2) by adenoviral vectors. Alterations in the fibrous actin (F-actin) content and organization and the expression of ß-catenin were detected by flow cytometry, immunofluorescence, and Western blotting. Additionally, the association of DACT1 with gap junctions connexin 43 (Cx43) was detected by immunohistochemistry, immunofluorescence, and Western blotting. RESULTS: Decreased cytoplasmic DACT1 expression in the myocardium was associated with AF (P=0.037) and a high degree of fibrosis (weak vs. strong, P=0.028; weak vs. very strong, P=0.029). A positive association was observed between DACT1 and ß-catenin expression in clinical samples (P=0.028, Spearman's rho=0.408). Furthermore, overexpression of DACT1 in HL-1 and H9C2 cells induced an increase in ß-catenin and subsequent partial colocalization of DACT1 and ß-catenin. In addition, F-actin content and organization were enhanced. Interestingly, DACT1 was positively correlated with the Cx43 expression in clinical samples (P=0.048, Spearman's rho=0.370) and changed the Cx43 distribution in cardiac cell lines. CONCLUSION: DACT1 proved to be a novel AF-related gene by regulating Cx43 via cytoskeletal organization induced by ß-catenin accumulation in cardiomyocytes. DACT1 could thus serve as a potential therapeutic marker for AF.

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Resumen Introducción. La leptina es una hormona secretada por los adipocitos que se ha relacionado con el proceso de la transición de epitelio a mesénquima (Epithelial- Mesenchymal Transition, EMT). Promueve la migración e invasión de las células del epitelio mamario mediante la activación de las cinasas FAK y Src, un complejo regulador de vías de señalización que favorecen la expresión de las proteínas relacionadas con la formación de estructuras proteolíticas implicadas en la invasión y progresión del cáncer. Recientemente, se ha descrito que la sobreexpresión y activación de la proteína Hic-5 durante el mencionado proceso de transición, favorece la formación de los puntos de actina (indicativa de la formación y funcionalidad de los invadopodios), lo cual promueve la degradación local de los componentes de la matriz extracelular y la metástasis del cáncer. Objetivos. Evaluar el papel de las cinasas FAK y Src sobre la expresión y localización subcelular de Hic-5 y la formación de puntos de actina inducida por la leptina en la línea celular MCF10A de epitelio mamario no tumoral. Materiales y métodos. Se utilizaron los inhibidores específicos de la FAK (PF-573228) y la Src (PP2) para evaluar el papel de ambas cinasas en los niveles de expresión y localización subcelular de la proteína Hic-5 mediante Western blot e inmunofluorescencia, así como la formación de puntos de actina mediante la tinción con faloidina-TRITC en células MCF10A estimuladas con leptina. Resultados. La leptina indujo el incremento en la expresión de Hic-5 y la formación de puntos de actina. El tratamiento previo con los inhibidores de las cinasas FAK (PF-573228) y Src (PP2), promovió la disminución en la expresión de Hic-5 y de los puntos de actina en la línea celular MCF10A de epitelio mamario no tumoral. Conclusión. La leptina indujo la expresión y la localización perinuclear de Hic-5 y la formación de puntos de actina mediante un mecanismo dependiente de la actividad de las cinasas FAK y Src en las células MCF10A.

Abstract Introduction: Leptin is a hormone secreted by adipocytes that has been associated with the epithelial-mesenchymal transition (EMT). Additionally, leptin promotes the migration and invasion of mammary epithelial cells through the activation of FAK and Src kinases, which are part of a regulatory complex of signaling pathways that promotes the expression of proteins related to the formation of proteolytic structures involved in the invasion and progression of cancer. Recently, overexpression and activation of Hic-5 during the EMT have been shown to induce the formation of actin puncta; these structures are indicative of the formation and functionality of invadopodia, which promote the local degradation of extracellular matrix components and cancer metastasis. Objective: To evaluate the role of FAK and Src kinases in the expression of Hic-5 during the epithelial-mesenchymal transition induced by leptin in MCF10A cells. Materials and methods: We used specific inhibitors of FAK (PF-573228) and Src (PP2) to evaluate Hic-5 expression and subcellular localization by Western blot and immunofluorescence assays and to investigate the formation of actin puncta by epifluorescence in MCF10A cells stimulated with leptin. Results: Leptin induced an increase in Hic-5 expression and the formation of actin puncta. Pretreatment with inhibitors of FAK (PF-573228) and Src (PP2) promoted a decrease in Hic-5 expression and actin puncta formation in the non-tumorigenic mammary epithelial cell line MCF10A. Conclusion: In MCF10A cells, leptin-induced Hic-5 expression and perinuclear localization, as well as the formation of actin puncta through a mechanism dependent on the kinase activity of FAK and Src.

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ABSTRACT α-smooth muscle actin, encoded by ACTA2 gene, is an isoform of the vascular smooth muscle actins, typically expressed in the vascular smooth muscle cells contributing to vascular motility and contraction. ACTA2 gene mutations cause a diversity of diffuse vasculopathies such as thoracic aortic aneurysms and dissections as well as occlusive vascular diseases, including premature coronary artery disease and ischemic stroke. Dynamics of differentiation-specific α-smooth muscle actin in arterial smooth muscle cells and proliferation of the proteins have been well described. Although a variety of research works have been undertaken in terms of modifications of α-smooth muscle actin and mutations of ACTA2 gene and myosin, the underlying mechanisms towards the pathological processes by way of gene mutations are yet to be clarified. The purpose of the present article is to describe the phenotypes of α-smooth muscle actin and implications of ACTA2 mutations in vasculopathies in order to enhance the understanding of potential mechanisms of aortic and coronary disorders.

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Introdução: Para o adequado estudo anatomopatológico dos tecidos obtidos dos pacientes é crucial a boa técnica para a melhor evidenciação das células pesquisadas. É imprescindível que não só a técnica ideal de coloração/marcação seja utilizada, mas também a correta técnica de fixação da peça cirúrgica. Em trabalho prévio pesquisando miofibroblastos e contração na pele restaurada de áreas doadoras de enxertos de pele de espessura parcial, verificamos baixa densidade de miofibroblastos, ao contrário dos estudos em tecido de granulação. Investigando as causas disso, e baseando-se em artigo específico sobre a eficácia da evidenciação imuno-histoquímica com diferentes métodos de fixação tecidual, comparamos a eficácia dos diferentes fixadores, formol a 10% e álcool a 70% para a evidenciação de miofibroblastos em tecido de granulação de úlceras crônicas de 21 pacientes. Método: Estudamos 42 amostras por meio de técnica imuno-histoquímica, utilizando-se o anticorpo anti-Smooth Muscle Actin, nas quais contamos os miofibrobastos em 10 campos com aumento de 400 X. Resultados: Demonstraram-se melhores resultados qualitativo e quantitativo no material fixado em álcool, quando comparado ao fixado em formol, com densidades médias de miofibroblastos de 86,5 e 48 células/mm2, respectivamente, o que nos leva a recomendar o álcool como fixador de tecidos para estas análises imuno-histoquímicas.

Background: The use of a good visualization technique is crucial for an adequate pathological anatomy study of tissues obtained from patients. Optimal staining and fixation techniques of the surgical piece are essential. In a previous study investigating myofibroblasts and contraction in split skingraft donor sites, a low density of myofibroblasts was observed, as opposed to studies in granulation studies. To investigate the causes for this finding and based on a specific manuscript on the efficacy of immunohistochemical visualization using different tissue fixation methods, we compared the efficacy of fixation in 10% formaldehyde and 70% alcohol for the visualization of myofibroblasts in granulation tissues of chronic ulcers in 21 patients. Method: Forty-two samples were studied by immunohistochemistry, using the anti-Smooth Muscle Actin antibody, and myofibroblasts were counted in 10 fields with a magnification of 400X. Results: Better qualitative and quantitative results were observed in alcohol-fixed materials, when compared to those fixed in formaldehyde, with mean myofibroblasts densities of 86.5 and 48 cells/mm2, respectively, which encourages us to recommend the use of alcohol for tissue fixation in immunohistochemical studies.

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