Evaluación del grado de queratinización y el recuento de AgNORs en citología exfoliativa de mucosa oral normal de individuos fumadores y no fumadores

Objetivos. En individuos fumadores con mucosa oral clínicamente sana, se han observado cambios citológicos como una mayor queratinización, existiendo también reportes de un mayor grado de actividad nucleolar. En estos estudios, las células para frotis se han obtenido por medio de espátula de madera. Nuestro objetivo es evaluar la profundidad de muestras citológicas de mucosa oral obtenidas con cepillo para frotis (endobrush) y comparar el grado de queratinización y la actividad nucleolar en pacientes fumadores y no fumadores. Diseño del estudio. Se obtuvieron frotis de mucosa oral de borde de lengua clínicamente normal de 30 individuos fumadores y 30 no fumadores, utilizando espátula de madera y endobrush. Las muestras fueron teñidas con Papanicolaou y con la tinción AgNORs. Resultados. Con la espátula de madera se obtuvo un mayor porcentaje de células epiteliales superficiales anucleadas (P= 0.016) y con el endobrush se obtuvieron células más profundas (tipo intermedias) (P= 0.035). Los individuos fumadores presentaron un mayor porcentaje de células superficiales anucleadas con ambas técnicas, diferencia que fue estadísticamente significativa con la técnica endobrush (P=0.005). El promedio de AgNORs en las células nucleadas fue mayor en los individuos fumadores (3.83) que en los no fumadores (2.79) (P= 0.003). Conclusiones. El endobrush permite obtener células de estratos más profundos. Los individuos fumadores con mucosa clínicamente normal presentan un mayor porcentaje de células queratinizadas y una mayor actividad nucleolar, sugiriendo que el consumo de cigarrillo influye en la actividad celular de la mucosa del borde de lengua.Objetive.In smokers with clinically normal buccal mucosa, cytological changes such as increased keratinization, and higher nucleolar activity have been observed. In these studies the cells for cytological smears were obtained with a wooden spatula. Our objectives were to evaluate the depth of cytological smears of oral mucosa obtained with both a brush (endobrush) and a wooden spatula, and to compare the degree of keratinization and the nucleolar activity in smokers and non-smokers. Design. We obtained cytological smears of clinically normal lateral tongues of 30 smokers and 30 non-smokers using both a wooden spatula and endobrush. The samples were dyed with Papanicolaou and the AgNORs. Results. With the wooden spatula we found a greater percentage of enucleated superficial epithelial cells (P = 0.016) and deeper cells were obtained with an endobrush (intermediate cells) (P =0.035). The smokers showed a greater percentage of enucleated superficial cells with both techniques, however this difference was significantly greater with Endobrush (P=0.005). The average of AgNORs in the nucleated cells was greater in smokers (3.83) than in non-smokers (2.79) (P=0.003). Conclusion. The Endobrush allows the clinician to obtain deeper cells of buccal mucosa. Smokers with clinically normal mucosa show a greater percentage of keratinized cells and a greater nucleolar activity, suggesting that cigarette smoking influences the cellular activity of the mucosa of the lateral tongue

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field