Ultraestructura de las lesiones producidas por antibióticos aminoglucósidos (kalamicina y gentamicina) en el coclea de cobaya

Tesis Univ. Complutense.Depto. de Medicina Legal, Psiquiatría y PatologíaFac. de MedicinaTRUEProQuestpu

Ultraestructura de las lesiones producidas por antibióticos aminoglucósidos (kalamicina y gentamicina) en el coclea de cobaya

Tesis Univ. Complutense.Depto. de Medicina Legal, Psiquiatría y PatologíaFac. de MedicinaTRUEProQuestpu

Microscopía digital como herramienta para la docencia práctica en Anatomía Patológica

Memoria ID2022-185 Ayudas de la Universidad de Salamanca para la innovación docente, curso 2022-2023

Clinical, genetic and pharmacological data support targeting the MEK5/ERK5 module in lung cancer

Article number: 78 (2021)[EN]Despite advances in its treatment, lung cancer still represents the most common and lethal tumor. Because of that, efforts to decipher the pathophysiological actors that may promote lung tumor generation/progression are being made, with the final aim of establishing new therapeutic options. Using a transgenic mouse model, we formerly demonstrated that the sole activation of the MEK5/ERK5 MAPK route had a pathophysiological role in the onset of lung adenocarcinomas. Given the prevalence of that disease and its frequent dismal prognosis, our findings opened the possibility of targeting the MEK5/ERK5 route with therapeutic purposes. Here we have explored such possibility. We found that increased levels of MEK5/ERK5 correlated with poor patient prognosis in lung cancer. Moreover, using genetic as well as pharmacological tools, we show that targeting the MEK5/ERK5 route is therapeutically effective in lung cancer. Not only genetic disruption of ERK5 by CRISPR/Cas9 caused a relevant inhibition of tumor growth in vitro and in vivo; such ERK5 deficit augmented the antitumoral effect of agents normally used in the lung cancer clinic. The clinical correlation studies together with the pharmacological and genetic results establish the basis for considering the targeting of the MEK5/ERK5 route in the therapy for lung cancer.Cancer Center Network Program from the ISCIII (RD12/0036/0003

How to improve the precision of Bethesda System diagnostic categories I and III

Fine-needle aspiration (FNA) has a primary role in the diagnosis of thyroid nodules. FNA allows us to differentiate benign lesions from malignant ones in most of the cases. However, management of Bethesda System diagnostic categories I (Nondiagnostic or Unsatisfactory) and III (Atypia of Undetermined Significance or Follicular Lesion of Undetermined Significance) are a clinical challenge./nThe aim of this work is to expose the options we have to improve the diagnosis of those categories using a multidisciplinary approach, an active follow-up, repeating FNA, applying immunohistochemical and/or molecular technics or even perform a core needle biopsy./nSome or all of these choices are especially useful to identify category III, but active follow-up or additional FNA are preferred to detect category I.Introducción y objetivo: La punción aspiración con aguja fina es el método de elección para el diagnóstico del nódulo tiroideo, permitiendo discriminar entre lesiones malignas y benignas en un alto porcentaje de pacientes. Sin embargo, las categorías diagnósticas I (No diagnóstico o insatisfactorio) y III (Atipia o Lesión folicular de Significado Incierto) del Sistema Bethesda suponen un reto en el manejo de los nódulos tiroideos. El objetivo del presente trabajo es exponer las opciones de que se disponen en el momento actual para intentar incrementar la rentabilidad de estos diagnósticos. Síntesis: En primer lugar, el abordaje multidisciplinar de los casos, la posibilidad de seguimiento, la repetición de la punción aspiración, la complementación del diagnóstico incorporando técnicas inmunohistoquímicas o moleculares, y finalmente la posibilidad de realizar una biopsiacilindro. Conclusiones: Estas alternativas son especialmente útiles en la categoría diagnóstica III, mientras que se prefiere la opción de seguimiento o repetición de PAAF en la categoría diagnóstica I

MEK5 promotes lung adenocarcinoma

[EN]Lung cancer represents the leading cause of cancer death worldwide [1]. Because of that, intense efforts are being devoted to the development of novel therapeutic strategies to fight the disease [2]. In this respect, identification of new oncogenic drivers offers therapeutic opportunities in tumours in which those molecules or other cooperating elements play a pathophysiological role. Here, we show that the MEK5 mitogen-activated protein kinase kinase has a pivotal role in lung cancer. Originally, this study was initiated with the purpose of evaluating the potential oncogenic role of the MEK5 pathway. In fact, while the MEK5 pathway has been found to be deregulated in several neoplasias [3–6], whether exclusive activation of that pathway promotes tumorigenesis has not previously been addressed. To explore that possibility, we generated transgenic mice engineered to express a constitutively active form of MEK5 by site-directed mutagenesis of the MEK5 Ser311 and Thr315 residues to aspartic acid (MEK5DD) (figure 1a). These acidic amino acid changes result in a MEK5 form in which the aspartic acid substitutions function as phosphomimetic residues [7, 8]. As a consequence, MEK5DD acts as a constitutively active kinase that is able to phosphorylate its downstream target, the ERK5 mitogen-activated protein kinase. Phosphorylation of ERK5 by constitutively active MEK5DD results in sustained activation of ERK5. Such ERK5 phosphorylation ( pERK5) provokes a change in its electrophoretic mobility with respect to unphosphorylated ERK5, a characteristic that can be used to differentiate ERK5 from pERK5 by Western blotting [9]. The MEK5DD cDNA was subcloned into the pCEFL mammalian expression vector, which contains an N-terminal Flag tag sequence that serves to differentiate MEK5DD from endogenous MEK5. Increasing amounts of the cDNA coding for Flag-tagged MEK5DD were transfected in HeLa cells and its expression was analysed by Western blotting with an anti-Flag antibody. As shown in figure 1b, expression of Flag-MEK5DD caused the appearance of pERK5, indicative of pathway activationGrants from the Instituto de Salud Carlos III (ISCIII) (PS09/00868 and PI15/01180) and by the Scientific Foundation of the Spanish Association Against Cancer (AECC)

T-cell intracellular antigens function as tumor suppressor genes

Knockdown of T-cell intracellular antigens TIA1 and TIAR in transformed cells triggers cell proliferation and tumor growth. Using a tetracycline-inducible system, we report here that an increased expression of TIA1 or TIAR in 293 cells results in reduced rates of cell proliferation. Ectopic expression of these proteins abolish endogenous TIA1 and TIAR levels via the regulation of splicing of their pre-mRNAs, and partially represses global translation in a phospho-eukaryotic initiation factor 2 alpha-dependent manner. This is accompanied by cell cycle arrest at G1/S and cell death through caspase-dependent apoptosis and autophagy. Genome-wide profiling illustrates a selective upregulation of p53 signaling pathway-related genes. Nude mice injected with doxycycline-inducible cells expressing TIA1 or TIAR retard, or even inhibit, growth of xenotumors. Remarkably, low expressions of TIA1 and TIAR correlate with poor prognosis in patients with lung squamous cell carcinoma. These findings strongly support the concept that TIA proteins act as tumor suppressor genes.Spanish Ministry of Economic Affairs and Competitiveness through FEDER funds (BFU2008-00354 and BFU2011-29653 to JMI) and the Junta de Castilla y León (GRS 600/A/11 to MDL). The CBMSO received an institutional grant from Fundación Ramón Arece

Factors influencing malignant evolution and long-term survival in solitary fibrous tumours of the pleura

Solitary pleuro-pulmonary fibrous tumours are relatively uncommon neoplasms that are difficult to manage therapeutically and which, cytogenetically, have been poorly studied. The aim of the present work was to analyse the characteristics of a series of consecutive operated solitary pleural fibrous tumours in an attempt to discover a malignant pattern of evolution. This was a retrospective observational study of 19 cases. Samples were studied for clinical, histological, immunohistochemical and cytogenetic characteristics (aCGH, FISH). Descriptive statistics were used: the Kapplan-Meyer log-rank test and the Cox-regression model for survival analysis. Analysis of malignant evolution was achieved using 2x2 tables; significant factors were included in a binary logistic regression model. Parietal pleural implantation of the primary tumour, high mib1 expression, and low p53 expression were seen to be statistically significant factors for survival. We recommend a close follow-up for patients with a malignant primary tumour and low p53 expression and a regular long-term follow-up for benign primary tumours with a high mib1 index, high positive p53, and deletions. These findings need confirmation in more extensive series.Peer Reviewe

Knockdown of T-cell intracellular antigens triggers cell proliferation, invasion and tumour growth

TIA (T-cell intracellular antigen) proteins function as DNA/RNA trans-acting regulators to expand transcriptome and proteome diversity in mammals. In the present paper we report that the stable silencing of TIA1 and/or TIAR/TIAL1 (TIA1-related/like protein 1) expression in HeLa cells enhances cell proliferation, anchorage-dependent and -independent growth and invasion. HeLa cells lacking TIA1 and/or TIAR generate larger and faster-growing epithelial tumours with high rates of proliferation and angiogenesis in nude mice xenografts. Protein array analysis of a collection of human tumours shows that TIA1 and TIAR protein expression is down-regulated in a subset of epithelial tumours relative to normal tissues. Our results suggest a link between the epigenetic control exerted by TIA proteins and the transcriptional and post-transcriptional regulation of a subset of specific genes involved in tumour progression. Taken together, these results are consistent with a role for TIA proteins as growth/tumour-suppressor genes.This work was supported by the Fondo de Investigaciones Sanitarias-FEDER [grant number PI051605]; the Ministerio de Ciencia e Innovación-FEDER [grant number BFU2008–00354]; and the Agencia Estatal Consejo Superior de Investigaciones Científicas [grant number 200920I108]. The Centro de Biología Molecular ‘Severo Ochoa’ receives an institutional grant from the Fundación Ramón Areces, Spain.Peer reviewe

Knockdown of T-cell intracellular antigens triggers cell proliferation, invasion and tumor growth.

International audienceT-cell intracellular antigen (TIA) proteins function as DNA/RNA trans-acting regulators to expand transcriptome and proteome diversity in mammals. We hereby report that stable silencing of TIA1 and/or TIAR/TIAL1 expression in HeLa cells enhances cell proliferation, anchorage-dependent and -independent growth and invasion. HeLa cells lacking TIA1 and/or TIAR generate larger and faster-growing epithelial tumors with high rates of proliferation and angiogenesis in nude mice xenografts. Protein array analysis of a collection of human tumors shows that TIA1 and TIAR protein expression is downregulated in a subset of epithelial tumors relative to normal tissues. Our data suggest a link between the epigenetic control exerted by TIA proteins on transcriptional and post-transcriptional regulation of a subset of specific gene involved in tumor progression. Taken together, these results are consistent with a role for TIA proteins as growth/tumor suppressor genes