Expresión de E-cadherina, laminina y colágeno IV en la evolución de displasia a carcinoma epidermoide oral

Objetivos: Estudiar la pérdida o reducción de la adhesión celular mediada por E-cadherina en leucoplasias, carcinomas epidermoides y metástasis ganglionares. Estudiar la pérdida de continuidad de la expresión de laminina y colágeno IV en la membrana basal epitelial en el desarrollo biológico de las leucoplasias y carcinomas orales. Material y metodo: Hemos estudiado 124 muestras de pacientes portadores de leucoplasias y carcinomas orales con diversos diagnósticos que abarcan desde epitelio normal (13 muestras), displasias leves (2), displasias moderadas (12), carcinomas in situ (13) carcinomas microinvasores (11) Carcinoma epidermoide oral (64 muestras) y metástasis ganglionar (9). Se construyeron 7 bloques de tissue microarrays con aguja de 2mm y se realizó un estudio mediante técnica inmunohistoquímica para E-cadherina (clona 36, T.D. ABD Company), Laminina (078P, Biogenex) y Colágeno IV (PHM12, Biogenex). Resultados: En Displasias Leves y Moderadas presentan pérdida de expresión de E-cadherina, Laminina, y Colágeno IV (20%). En Carcinomas in situ y Microinvasores, presentaron pérdida de expresión de E-cadherina (73%), y en Laminina y Colágeno IV (57%). En los carcinomas epidermoides, encontramos pérdida de expresión de E-cadherina (90%) y discontinuidad en la M. basal (70%). Todas las metástasis ganglionares presentaron pérdida de E-cadherina y discontinuidad en Laminina y Colágeno IV. Conclusiones: La pérdida de expresión de E-cadherina se incrementa al aumentar el grado de displasia de las lesiones. La perdida de continuidad en la expresión de laminina y Colágeno IV sigue una evolución paralela desde displasias a metástasis ganglionares. La disminución en la expresión de los tres marcadores ha sido significativa en la evolución de las lesiones orales.Objectives: Study the loss or reduction of the cellular adhesion mediated for E-cadherin in oral leukoplakias, oral squamous cell carcinomas and metastatic nodules. Study the loss of continuity of the laminin and collagen IV expression in the epithelial basal membrane from the biological development of the oral leukoplakias and oral carcinomas. Material and method: we have studied 124 samples of patient payees leukoplakias and oral carcinomas with diverse diagnosis that embrace from normal epithelium (13 samples), mild dysplasias (2), moderate dysplasias (12), 'in situ' carcinomas (13), microinvasive carcinomas (11) oral squamous cell carcinomas (64 samples) and metastatic nodules (9). 7 blocks of tissue microarrays were built with needle of 2mm and was carried out a study by means of immunohistochemical technique for E-cadherin (clone 36, Biogenex), Laminin (078P, Biogenex) and Collagen IV (PHM12, Biogenex). Results: In Mild and Moderate Dysplasias the results present loss of E-cadherin, Laminin, and Collagen IV (20%) expression. 'in situ' and microinvasive carcinomas, the results presented loss of E-cadherin expression (73%), and loss in Laminin and Collagen IV expression (57%). In the squamous cell carcinomas , we find E-cadherin underexpression (90%) and discontinuity in the Basal Membrane. (70%). All the metastatic nodules presented loss of E-cadherin expression and discontinuity in Laminin and Collagen IV expression. Conclusions: The loss of E-cadherin expression is increased when increasing the dysplasia grade of lesions. The loss of continuity in the laminin and Collagen IV expression follow a parallel evolution from dysplasias to metastatic nodules. The underexpression of the three markers has been significant in the evolution of the oral lesions

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

E-cadherin, laminin and collagen IV expression in the evolution from dysplasia to oral squamous cell carcinoma

Objetivos: Estudiar la pérdida o reducción de la adhesión celular mediada por E-cadherina en leucoplasias, carcinomas epidermoides y metástasis ganglionares. Estudiar la pérdida de continuidad de la expresión de laminina y colágeno IV en la membrana basal epitelial en el desarrollo biológico de las leucoplasias y carcinomas orales. Material y metodo: Hemos estudiado 124 muestras de pacientes portadores de leucoplasias y carcinomas orales con diversos diagnósticos que abarcan desde epitelio normal (13 muestras), displasias leves (2), displasias moderadas (12), carcinomas in situ (13) carcinomas microinvasores (11) Carcinoma epidermoide oral (64 muestras) y metástasis ganglionar (9). Se construyeron 7 bloques de tissue microarrays con aguja de 2mm y se realizó un estudio mediante técnica inmunohistoquímica para E-cadherina (clona 36, T.D. ABD Company), Laminina (078P, Biogenex) y Colágeno IV (PHM12, Biogenex). Resultados: En Displasias Leves y Moderadas presentan pérdida de expresión de E-cadherina, Laminina, y Colágeno IV (20%). En Carcinomas in situ y Microinvasores, presentaron pérdida de expresión de E-cadherina (73%), y en Laminina y Colágeno IV (57%). En los carcinomas epidermoides, encontramos pérdida de expresión de E-cadherina (90%) y discontinuidad en la M. basal (70%). Todas las metástasis ganglionares presentaron pérdida de E-cadherina y discontinuidad en Laminina y Colágeno IV. Conclusiones: La pérdida de expresión de E-cadherina se incrementa al aumentar el grado de displasia de las lesiones. La perdida de continuidad en la expresión de laminina y Colágeno IV sigue una evolución paralela desde displasias a metástasis ganglionares. La disminución en la expresión de los tres marcadores ha sido significativa en la evolución de las lesiones orales

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)

Spatio-temporal tumor heterogeneity in metastatic CRC tumors: a mutational-based approach

[EN] It is well known that activating mutations in the KRAS and NRAS genes are associated with poor response to anti-EGFR therapies in patients with metastatic colorectal cancer (mCRC). Approximately half of the patients with wild-type (WT) KRAS colorectal carcinoma do not respond to these therapies. This could be because the treatment decision is determined by the mutational profile of the primary tumor, regardless of the presence of small tumor subclones harboring RAS mutations in lymph nodes or liver metastases. We analyzed the mutational profile of the KRAS, NRAS, BRAF and PI3KCA genes using low-density microarray technology in samples of 26 paired primary tumors, 16 lymph nodes and 34 liver metastases from 26 untreated mCRC patients (n=76 samples). The most frequent mutations found in primary tumors were KRAS (15%) and PI3KCA (15%), followed by NRAS (8%) and BRAF (4%). The distribution of the mutations in the 16 lymph node metastases analyzed was as follows: 4 (25%) in KRAS gene, 3 (19%) in NRAS gene and 1 mutation each in PI3KCA and BRAF genes (6%). As expected, the most prevalent mutation in liver metastasis was in the KRAS gene (35%), followed by PI3KCA (9%) and BRAF (6%). Of the 26 cases studied, 15 (58%) displayed an overall concordance in the mutation status detected in the lymph node metastases and liver metastases compared with primary tumor, suggesting no clonal evolution. In contrast, the mutation profiles differed in the primary tumor and lymph node/metastases samples of the remaining 11 patients (48%), suggesting a spatial and temporal clonal evolution. We confirm the presence of different mutational profiles among primary tumors, lymph node metastases and liver metastases. Our results suggest the need to perform mutational analysis in all available tumor samples of patients before deciding to commence anti-EGFR treatment

Cualificación en los Objetivos establecidos en la Agenda 2030 de estudiantes y profesores en el Máster Universitario en Profesor de Educación Secundaria Obligatoria y Bachillerato, Formación Profesional y Enseñanza de Idiomas (MUPES)

Memoria ID2022-157 Ayudas de la Universidad de Salamanca para la innovación docente, curso 2022-2023

Role of age and comorbidities in mortality of patients with infective endocarditis

[Purpose]: The aim of this study was to analyse the characteristics of patients with IE in three groups of age and to assess the ability of age and the Charlson Comorbidity Index (CCI) to predict mortality. [Methods]: Prospective cohort study of all patients with IE included in the GAMES Spanish database between 2008 and 2015.Patients were stratified into three age groups:<65 years,65 to 80 years,and ≥ 80 years.The area under the receiver-operating characteristic (AUROC) curve was calculated to quantify the diagnostic accuracy of the CCI to predict mortality risk. [Results]: A total of 3120 patients with IE (1327 < 65 years;1291 65-80 years;502 ≥ 80 years) were enrolled.Fever and heart failure were the most common presentations of IE, with no differences among age groups.Patients ≥80 years who underwent surgery were significantly lower compared with other age groups (14.3%,65 years; 20.5%,65-79 years; 31.3%,≥80 years). In-hospital mortality was lower in the <65-year group (20.3%,<65 years;30.1%,65-79 years;34.7%,≥80 years;p < 0.001) as well as 1-year mortality (3.2%, <65 years; 5.5%, 65-80 years;7.6%,≥80 years; p = 0.003).Independent predictors of mortality were age ≥ 80 years (hazard ratio [HR]:2.78;95% confidence interval [CI]:2.32–3.34), CCI ≥ 3 (HR:1.62; 95% CI:1.39–1.88),and non-performed surgery (HR:1.64;95% CI:11.16–1.58).When the three age groups were compared,the AUROC curve for CCI was significantly larger for patients aged <65 years(p < 0.001) for both in-hospital and 1-year mortality. [Conclusion]: There were no differences in the clinical presentation of IE between the groups. Age ≥ 80 years, high comorbidity (measured by CCI),and non-performance of surgery were independent predictors of mortality in patients with IE.CCI could help to identify those patients with IE and surgical indication who present a lower risk of in-hospital and 1-year mortality after surgery, especially in the <65-year group

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Effectiveness of an intervention for improving drug prescription in primary care patients with multimorbidity and polypharmacy:Study protocol of a cluster randomized clinical trial (Multi-PAP project)

This study was funded by the Fondo de Investigaciones Sanitarias ISCIII (Grant Numbers PI15/00276, PI15/00572, PI15/00996), REDISSEC (Project Numbers RD12/0001/0012, RD16/0001/0005), and the European Regional Development Fund ("A way to build Europe").Background: Multimorbidity is associated with negative effects both on people's health and on healthcare systems. A key problem linked to multimorbidity is polypharmacy, which in turn is associated with increased risk of partly preventable adverse effects, including mortality. The Ariadne principles describe a model of care based on a thorough assessment of diseases, treatments (and potential interactions), clinical status, context and preferences of patients with multimorbidity, with the aim of prioritizing and sharing realistic treatment goals that guide an individualized management. The aim of this study is to evaluate the effectiveness of a complex intervention that implements the Ariadne principles in a population of young-old patients with multimorbidity and polypharmacy. The intervention seeks to improve the appropriateness of prescribing in primary care (PC), as measured by the medication appropriateness index (MAI) score at 6 and 12months, as compared with usual care. Methods/Design: Design:pragmatic cluster randomized clinical trial. Unit of randomization: family physician (FP). Unit of analysis: patient. Scope: PC health centres in three autonomous communities: Aragon, Madrid, and Andalusia (Spain). Population: patients aged 65-74years with multimorbidity (≥3 chronic diseases) and polypharmacy (≥5 drugs prescribed in ≥3months). Sample size: n=400 (200 per study arm). Intervention: complex intervention based on the implementation of the Ariadne principles with two components: (1) FP training and (2) FP-patient interview. Outcomes: MAI score, health services use, quality of life (Euroqol 5D-5L), pharmacotherapy and adherence to treatment (Morisky-Green, Haynes-Sackett), and clinical and socio-demographic variables. Statistical analysis: primary outcome is the difference in MAI score between T0 and T1 and corresponding 95% confidence interval. Adjustment for confounding factors will be performed by multilevel analysis. All analyses will be carried out in accordance with the intention-to-treat principle. Discussion: It is essential to provide evidence concerning interventions on PC patients with polypharmacy and multimorbidity, conducted in the context of routine clinical practice, and involving young-old patients with significant potential for preventing negative health outcomes. Trial registration: Clinicaltrials.gov, NCT02866799Publisher PDFPeer reviewe