Generation, Expansion, and Biobanking of Gastrointestinal Patient-Derived Organoids from Tumor and Normal Tissues.

Patient-derived organoids (PDOs) generated from adult stem cells present in tissues are invaluable tools for translational cancer research (Drost, Clevers, Nat Rev Cancer 18(7):407-418, 2018). The generation of this 3D cultures is not trivial and requires dedicated procedures. Despite the rapid increase in the use of organoids in cancer research, it is noteworthy that published procedures regarding their generation often lack critical information and standardized protocols remain elusive. Addressing these limitations, the protocol described in this chapter offers an in-depth and comprehensive guide to establishing, expanding, and freezing gastrointestinal PDOs obtained from normal and tumor tissue biopsies. Notably, it also provides valuable insights in the form of tips and tricks to guide and overcome potential challenges that may arise during the procedure.

How organoids can improve personalized treatment in patients with gastro-esophageal tumors.

Gastro-esophageal tumors constitute a big health problem. Treatment options still mainly rely on chemotherapy, and apart from human epidermal growth factor receptor 2 positive and microsatellite instable/Epstein-Barr Virus disease, there are no molecularly guided options. Therefore, despite the large number of identified molecular alterations, precision medicine is still far from the clinic. In this context, the recently developed technology of patient-derived organoids (PDOs) could offer the chance to accelerate drug development and biomarker discovery. Indeed, PDOs are 3D primary cultures that were shown to reproduce patient's tumor characteristics. Moreover, several reports indicated that PDOs can replicate patient's response to a given drug; therefore, they are one of the most promising tools for functional precision medicine.

Estrategia de intervención con el candidato vacunal Abdala en trabajadores de la Salud

El proceso de vacunación contra la COVID-19 en Cuba está sustentado en fundamentos epistemológicos de la Medicina y las Ciencias biomédicas que permitieron la ejecución de los ensayos clínicos con vacunas de producción nacional, así como el desarrollo de otros candidatos vacunales durante el periodo de emergencia epidemiológica. El objetivo de este artículo es fundamentar los conocimientos en relación a la estrategia de intervención con el candidato vacunal Abdala en trabajadores de la Salud en Santiago de Cuba. Mediante una revisión bibliográfica realizada desde el 1ro. de marzo hasta el 31 de mayo de 2021, se analizaron artículos científicos, datos publicados por la Organización Mundial de la Salud, y libros de textos, todo lo cual facilitó la recuperación y valoración de la información sistematizada. La estrategia de intervención con el candidato vacunal Abdala en trabajadores de la salud en Santiago de Cuba demandó una gestión transdisciplinaria, intersectorial y participativa. La aplicación de los procedimientos del método clínico-epidemiológico reveló la necesidad de amplificar una cultura de inmunización con sustentos epistemológicos mediante la sistematización del proceso asistencial.

The vaccination process against COVID-19 in Cuba is based on epistemological foundations of Medicine and Biomedical Sciences that allowed the execution of clinical trials with nationally produced vaccines, as well as the development of other vaccine candidates during the epidemiological emergency period. The objective of this article is to base the knowledge in relation to the intervention strategy with the vaccine candidate Abdala in Health workers in Santiago de Cuba. Through a bibliographical review carried out from the 1st. From March to May 31st, 2021, scientific articles, data published by the World Health Organization, and textbooks were analyzed, all of which facilitated the recovery and evaluation of systematized information. The intervention strategy with the vaccine candidate Abdala in health workers in Santiago de Cuba demanded a transdisciplinary, intersectoral and participatory management. The application of the procedures of the clinical-epidemiological method revealed the need to amplify an immunization culture with epistemological supports through the systematization of the care process.

"Proteotranscriptomic analysis of advanced colorectal cancer patient derived organoids for drug sensitivity prediction".

BACKGROUND: Patient-derived organoids (PDOs) from advanced colorectal cancer (CRC) patients could be a key platform to predict drug response and discover new biomarkers. We aimed to integrate PDO drug response with multi-omics characterization beyond genomics. METHODS: We generated 29 PDO lines from 22 advanced CRC patients and provided a morphologic, genomic, and transcriptomic characterization. We performed drug sensitivity assays with a panel of both standard and non-standard agents in five long-term cultures, and integrated drug response with a baseline proteomic and transcriptomic characterization by SWATH-MS and RNA-seq analysis, respectively. RESULTS: PDOs were successfully generated from heavily pre-treated patients, including a paired model of advanced MSI high CRC deriving from pre- and post-chemotherapy liver metastasis. Our PDOs faithfully reproduced genomic and phenotypic features of original tissue. Drug panel testing identified differential response among PDOs, particularly to oxaliplatin and palbociclib. Proteotranscriptomic analyses revealed that oxaliplatin non-responder PDOs present enrichment of the t-RNA aminoacylation process and showed a shift towards oxidative phosphorylation pathway dependence, while an exceptional response to palbociclib was detected in a PDO with activation of MYC and enrichment of chaperonin T-complex protein Ring Complex (TRiC), involved in proteome integrity. Proteotranscriptomic data fusion confirmed these results within a highly integrated network of functional processes involved in differential response to drugs. CONCLUSIONS: Our strategy of integrating PDOs drug sensitivity with SWATH-mass spectrometry and RNA-seq allowed us to identify different baseline proteins and gene expression profiles with the potential to predict treatment response/resistance and to help in the development of effective and personalized cancer therapeutics.

Will Organoids Fill the Gap towards Functional Precision Medicine?

Precision medicine approaches for solid tumors are mainly based on genomics. Its employment in clinical trials has led to somewhat underwhelming results, except for single responses. Moreover, several factors can influence the response, such as gene and protein expression, the coexistence of different genomic alterations or post-transcriptional/translational modifications, the impact of tumor microenvironment, etc., therefore making it insufficient to employ a genomics-only approach to predict response. Recently, the implementation of patient-derived organoids has shed light on the possibility to use them to predict patient response to drug treatment. This could offer for the first time the possibility to move precision medicine to a functional environment.

Integrative immune transcriptomic classification improves patient selection for precision immunotherapy in advanced gastro-oesophageal adenocarcinoma.

BACKGROUND: Advanced gastro-oesophageal cancer (GEA) treatment has been improved by the introduction of immune checkpoint inhibitors (CPIs), yet identifying predictive biomarkers remains a priority, particularly in patients with a combined positive score (CPS) < 5, where the benefit is less clear. Our study assesses certain immune microenvironment features related to sensitivity or resistance to CPIs with the aim of implementing a personalised approach across CPS < 5 GEA. DESIGN: Through integrative transcriptomic and clinicopathological analyses, we studied in both a retrospective and a prospective cohort, the immune tumour microenvironment features. We analysed the cell types composing the immune infiltrate highlighting their functional activity. RESULTS: This integrative study allowed the identification of four different groups across our patients. Among them, we identified a cluster whose tumours expressed the most gene signatures related to immunomodulatory pathways and immunotherapy response. These tumours presented an enriched immune infiltrate showing high immune function activity that could potentially achieve the best benefit from CPIs. Finally, our findings were proven in an external CPI-exposed population, where the use of our transcriptomic results combined with CPS helped better identify those patients who could benefit from immunotherapy than using CPS alone (p = 0.043). CONCLUSIONS: This transcriptomic classification could improve precision immunotherapy for GEA.

Alternative Splicing, Epigenetic Modifications and Cancer: A Dangerous Triangle, or a Hopeful One?

The alteration of epigenetic modifications often causes cancer onset and development. In a similar way, aberrant alternative splicing may result in oncogenic products. These issues have often been individually reviewed, but there is a growing body of evidence for the interconnection of both causes of cancer. Actually, aberrant splicing may result from abnormal epigenetic signalization and epigenetic factors may be altered by alternative splicing. In this way, the interrelation between epigenetic marks and alternative splicing form the base of a triangle, while cancer may be placed at the vertex. The present review centers on the interconnections at the triangle base, i.e., between alternative splicing and epigenetic modifications, which may result in neoplastic transformations. The effects of different epigenetic factors, including DNA and histone modifications, the binding of non-coding RNAs and the alterations of chromatin organization on alternative splicing resulting in cancer are first considered. Other less-frequently considered questions, such as the epigenetic regulation of the splicing machinery, the aberrant splicing of epigenetic writers, readers and erasers, etc., are next reviewed in their connection with cancer. The knowledge of the above-mentioned relationships has allowed increasing the collection of biomarkers potentially useful as cancer diagnostic and/or prognostic tools. Finally, taking into account on one hand that epigenetic changes are reversible, and some epigenetic drugs already exist and, on the other hand, that drugs intended for reversing aberrations in alternative splicing, therapeutic possibilities for breaking the mentioned cancer-related triangle are discussed.

Evolución y pronóstico de pacientes con síndrome metabólico infectados por el nuevo coronavirus / Clinical course and prognosis of patients with metabolic syndrome infected by the new coronavirus

Introducción: La COVID-19 es una infección del tracto respiratorio causada por el SARS-CoV-2, que aparece en pacientes con elevado factores de riesgo como el síndrome metabólico. Objetivo: Actualizar los conocimientos sobre la evolución y el pronóstico de pacientes con síndrome metabólico infectados por el nuevo coronavirus. Desarrollo: Las fuentes secundarias y terciarias consultadas explican la relación directa entre los factores de riesgo del síndrome metabólico y las formas graves de la COVID-19, de manera que la evolución clínica y el pronóstico de estos pacientes es muy desfavorable, a pesar de que los protocolos terapéuticos de actuación hoy día se consideran complejos y contextualizados. Conclusiones: La desfavorable evolución clínica de los pacientes con síndrome metabólico infectados por la COVID-19 enmascara su pronóstico, por lo que las estrategias terapéuticas actuales para la atención a estos enfermos no han podido detener el curso progresivo de la enfermedad infecciosa identificada en la comorbilidad metabólica, lo que dificulta la prevención de complicaciones.

Introduction: The COVID-19 is an infection of the respiratory tract caused by the SARS-CoV-2 that appears in patients with high risk factors as the metabolic syndrome. Objective: To update the knowledge on the clinical course and prognosis of patients with metabolic syndrome infected by the new coronavirus. Development: The secondary and tetiary sources consulted explain the direct relationship between the risk factors of the metabolic syndrome and the serious forms of the COVID-19, so that the clinical course and prognosis of these patients is very unfavorable, although the performance therapeutic protocols nowadays are considered complex and contextualized. Conclusions: The unfavorable clinical course of the patients with metabolic syndrome infected by the COVID-19 hides its prognosis, reason why the current therapeutic strategies for the care to these sick persons have not been able to stop the progressive course of the infectious disease identified in the metabolic comorbidity, that makes difficult the prevention of complications.

Molecular profiling of advanced solid tumours. The impact of experimental molecular-matched therapies on cancer patient outcomes in early-phase trials: the MAST study.

INTRODUCTION: Molecular-matched therapies have revolutionized cancer treatment. We evaluated the improvement in clinical outcomes of applying an in-house customized Next Generation Sequencing panel in a single institution. METHODS: Patients with advanced solid tumors were molecularly selected to receive a molecular-matched treatment into early phase clinical trials versus best investigators choice, according to the evaluation of a multidisciplinary molecular tumor board. The primary endpoint was progression-free survival (PFS) assessed by the ratio of patients presenting 1.3-fold longer PFS on matched therapy (PFS2) than with prior therapy (PFS1). RESULTS: Of a total of 231 molecularly screened patients, 87 were eligible for analysis. Patients who received matched therapy had a higher median PFS2 (6.47 months; 95% CI, 2.24-14.43) compared to those who received standard therapy (2.76 months; 95% CI, 2.14-3.91, Log-rank p = 0.022). The proportion of patients with a PFS2/PFS1 ratio over 1.3 was significantly higher in the experimental arm (0.33 vs 0.08; p = 0.008). DISCUSSION: We demonstrate the pivotal role of the institutional molecular tumor board in evaluating the results of a customized NGS panel. This process optimizes the selection of available therapies, improving disease control. Prospective randomized trials are needed to confirm this approach and open the door to expanded drug access.

Epigenetic Mechanisms Are Involved in the Oncogenic Properties of ZNF518B in Colorectal Cancer.

The ZNF518B gene, which is up-regulated in colorectal cancer, plays a role in cell dissemination and metastasis. It encodes a zinc-finger protein, which interacts with histone methyltransferases G9A and EZH2. The expression of the two major mRNA isoforms 1 (coding for the full protein) and 2 was quantified by RT-qPCR in a cohort of 66 patients. The effects of silencing ZNF518B on the transcriptome of DLD1 and HCT116 cells were analysed by Clariom-S assays and validated by RT-qPCR. The recruitment of methyltransferases and the presence of H3K27me3 were studied by chromatin immunoprecipitation (ChIP). The ratio (isoform 2)/(isoform 1) negatively correlated with the relapsing of disease. The study of the transcriptome of DLD1 and HCT116 cells revealed that many genes affected by silencing ZNF518B are related to cancer. After crossing these results with the list of genes affected by silencing the histone methyltransferases (retrieved in silico), five genes were selected. ChIP analysis revealed that the recruitment of EZH2 is ZNF518B-dependent in KAT2B, RGS4 and EFNA5; the level of H3K27me3 changes in accordance. G9A also binds RGS4 and PADI3 in a ZNF518B-dependent manner. The results highlight the importance of epigenetics in cancer and open a novel therapeutic possibility, as inhibition of histone methyltransferases may reverse the disease-linked histone marks.

La educación médica en el diagnóstico educativo-terapéutico de pacientes hipertensos con rigidez arterial alterada / Medical education in the educational-therapeutic diagnosis of hypertensive patients with altered arterial stiffness

RESUMEN Fundamento: el carácter preventivo del sistema de salud cubano, como principio para su desarrollo, orienta la formación de un médico que responda consecuentemente a los problemas de salud de la población; de ahí que la educación médica los prepara para aplicar los últimos descubrimientos científicos en la prevención y tratamiento de enfermedades. Objetivo: valorar la contribución de una estrategia de diagnóstico educativo-terapéutico dirigido a la prevención de complicaciones vasculares que produce la hipertensión arterial. Métodos: se realizó una investigación pedagógica en el Policlínico Universitario "José Martí Pérez" de Santiago de Cuba durante 2019. Se emplearon métodos teóricos: análisis-síntesis, sistémico-estructural, holístico-dialéctico, hermenéutico-dialéctico e inductivo-deductivo; empíricos: encuestas, análisis documental, taller de socialización con especialistas, y técnicas educativas; estadísticos: técnicas descriptivas y análisis porcentual. Resultados: se aplicó una estrategia constituida por acciones de intervención educativo-terapéutica concretada en un proyecto de diagnóstico educativo de salud, la cual permitió incrementar el nivel de conocimientos de los pacientes hipertensos sobre su enfermedad; finalizada su aplicación se comprobó que no desarrollaron complicaciones vasculares lo que generó mayor compromiso con su salud. Conclusiones: los especialistas valoraron como adecuados el carácter científico profesional de la estrategia y su pertinencia. Su implementación contribuyó a la educación de los pacientes como un aporte de la educación médica en el nivel de atención primaria de salud.

ABSTRACT Background: the preventive nature of the Cuban health system, as a principle for its development, guides the training of a doctor who consistently responds to the health problems of the population; hence medical education prepares them to apply the latest scientific findings in disease prevention and treatment. Objective: to assess the contribution of an educational-therapeutic diagnostic strategy aimed at preventing vascular complications caused by hypertension. Methods: a pedagogical research was carried out at "José Martí Pérez" University Polyclinic in Santiago de Cuba during 2019. Theoretical methods were used: analysis-synthesis, systemic-structural, holistic-dialectical, hermeneutical-dialectical and inductive-deductive; empirical ones: surveys, documentary analysis, socialization workshop with specialists, and educational techniques; statistics: descriptive techniques and percentage analysis. Results: a strategy consisting of educational-therapeutic intervention actions was applied, it was concretized in a health educational diagnosis project, which allowed to increase the level of knowledge of hypertensive patients about their disease; after its implementation, it was found that they did not develop vascular complications, which generated a greater commitment to their health. Conclusions: the specialists valued the professional scientific nature of the strategy and its relevance as adequate. Its implementation contributed to the education of patients as a contribution to medical education at the primary health care level.

Detection of postoperative plasma circulating tumour DNA and lack of CDX2 expression as markers of recurrence in patients with localised colon cancer.

BACKGROUND: Colon cancer (CC) is a heterogeneous disease. Novel prognostic factors beyond pathological staging are required to accurately identify patients at higher risk of relapse. Integrating these new biological factors, such as plasma circulating tumour DNA (ctDNA), CDX2 staining, inflammation-associated cytokines and transcriptomic consensus molecular subtypes (CMS) classification, into a multimodal approach may improve our accuracy in determining risk of recurrence. METHODS: One hundred and fifty patients consecutively diagnosed with localised CC were prospectively enrolled in our study. ctDNA was tracked to detect minimal residual disease by droplet digital PCR. CDX2 expression was analysed by immunostaining. Plasma levels of cytokines potentially involved in disease progression were measured using ELISAs. A 96 custom gene panel for nCounter assay was used to classify CC into colorectal cancer assigner and CMS. RESULTS: Most patients were classified into CMS4 (37%) and CMS2 (28%), followed by CMS1 (20%) and CMS3 (15%) groups. CDX2-negative tumours were enriched in CMS1 and CMS4 subtypes. In univariable analysis, prognosis was influenced by primary tumour location, stage, vascular and perineural invasion together with high interleukin-6 plasma levels at baseline, tumours belonging to CMS 1 vs CMS2 +CMS3, ctDNA presence in plasma and CDX2 loss. However, only positive ctDNA in plasma samples (HR 13.64; p=0.002) and lack of CDX2 expression (HR 23.12; p=0.001) were found to be independent prognostic factors for disease-free survival in the multivariable model. CONCLUSIONS: ctDNA detection after surgery and lack of CDX2 expression identified patients at very high risk of recurrence in localised CC.

Prevención, diagnóstico y tratamiento de la sepsis neonatal: Guía de práctica clínica basada en evidencias del Instituto Nacional Materno Perinatal del Perú / Prevention, diagnosis and treatment of the neonatal sepsis: Clinical practice guideline-based evidence in a peruvian institute specialized

RESUMEN Introducción. La sepsis neonatal es un conjunto de signos y síntomas clínicos causados por una infección sistémica, asociada a factores de riesgo de tipo materno, neonatal u hospitalario. Objetivo. Brindar recomendaciones informadas por la mejor evidencia disponible para la prevención, diagnóstico y tratamiento de la sepsis neonatal. Métodos. Se desarrolló una guía de práctica clínica (GPC) basada en evidencias, mediante un proceso de adaptación, a cargo de un equipo de metodólogos y médicos neonatólogos expertos en el manejo clínico de la sepsis neonatal. Se realizó la búsqueda y preselección de GPC que respondan al alcance y objetivos planteados, utilizando el instrumento AGREE-II se evalúo la calidad metodológica de las guías para decidir su adaptación. Se realizó una búsqueda sistemática en múltiples bases de datos: Medline/PubMed, Embase/Ovid, Cochrane Library y LILACS, para identificar la evidencia que responda a las preguntas de la guía. Estas fueron seleccionadas y analizadas críticamente por pares clínicos y metodológicos, las recomendaciones fueron elaboradas usando el enfoque GRADE. Resultados. Se formularon 16 preguntas clínicas y recomendaciones basadas en evidencia a las que se llegó, mediante un diálogo deliberativo de expertos clínicos de diferentes hospitales de referencia para el manejo de la sepsis neonatal en el Perú. Las recomendaciones abordan la identificación de factores de riesgo, el uso de métodos confirmatorios y el tratamiento antibiótico como profilaxis y durante manejo de la enfermedad. Conclusiones. La GPC permite estandarizar el manejo clínico de la sepsis neonatal, así como la identificación de necesidades de investigación a realizarse en el contexto peruano.

ABSTRACT Introduction. Neonatal sepsis is a set of clinical signs and symptoms caused by a systemic infection, associated with maternal, neonatal or hospital risk factors. Objective. Provide informed recommendations for the best available evidence for the prevention, diagnosis and treatment of neonatal sepsis. Methods an Evidence-based Clinical Practice Guide (CPG) was developed through an adaptation process, in charge of a team of methodologists and neonatologists who are experts in the clinical management of neonatal sepsis. The search and preselection of CPGs that respond to the scope and objectives set were carried out, using the AGREE-II instrument, the methodological qualityof the guides was evaluated and their adaptation decided. To identify the evidence that answers the questions in the guideline, a systematic search was carried out in multiple databases: Medline / PubMed, Embase / Ovid, Cochrane Library and LILACS. These were selected and critically analyzed by clinical and methodological peers, the recommendations were elaborated using the GRADE approach. Results. 16 clinical questions and recommendations based on evidence were formulated, which were reached, through a deliberative dialogue of clinical experts from different reference hospitals for the management of neonatal sepsis in Peru. The recommendations address the identification of risk factors, the use of confirmatory methods and antibiotic treatment as prophylaxis and during disease management. Conclusions. The CPG allows standardizing the clinical management of neonatal sepsis, as well as the identification of research needs to be carried out in the Peruvian context.

Características clínico-epidemiológicas en pacientes con síndrome metabólico / Clinical and epidemiological characteristics of patients with metabolic syndrome

Introduction: The metabolic syndrome is defined by the presence of obesity, fundamentally central, hypertension, diabetes mellitus, and dyslipidemia; Objective: To characterize patients with metabolic syndrome from the clinical and epidemiological points of view. Methods: A descriptive and prospective study of 640 patients with metabolic syndrome was carried out, who were admitted to Dr. Juan Bruno Zayas Alfonso Teaching General Hospital from Santiago de Cuba during 2019. Information was statistically processed and analyzed; through the SPSS 11.5.1 program, by using the percentage as summary measure for qualitative variables. Results: In the serie, 26.8% of the patients belonged to the age group 40-49, and 53,0% were of female sex. Likewise, diabetes mellitus and hypertension were the associated diseases in 22,3 y 22,2 % of the patients, respectively, while 67,7 % had a saturated fats diet. Social drinkers were identified in 53,0 % of cases, and the heavy smokers in 47.0 %. Conclusions: Personal and family associated diseases prevailed as associated personal and family diseases, what shows the closed relationship between them and the metabolic syndrome. Health promotion and education should be considered in patients with metabolic syndrome from the primary care, as well as the distribution of medical care for this group of hospital institutions, to prevent and control risk factors related to its emergence.

Introducción: El síndrome metabólico está definido por la presencia de obesidad, fundamentalmente central; hipertensión arterial; diabetes mellitus y dislipidemias. Objetivo: Caracterizar a pacientes con síndrome metabólico desde el punto de vista clínico-epidemiológico. Métodos: Se realizó un estudio descriptivo y prospectivo de 640 pacientes con síndrome metabólico atendidos en el Hospital General Docente Dr. Juan Bruno Zayas Alfonso de Santiago de Cuba durante el año 2019. La información se procesó y analizó estadísticamente mediante el programa SPSS 11.5.1, con el empleo del porcentaje como medida de resumen para variables cualitativas. Resultados: En la serie, 26,8 % perteneció al grupo de edades de 40-49 años y 53,0 % era del sexo femenino. Asimismo, la diabetes mellitus y la hipertensión arterial figuraron como enfermedades asociadas en 22,3 y 22,2 % de los pacientes, respectivamente, en tanto 67,7 % ingería dieta a base de grasa saturada. Los bebedores sociales fueron identificados en 53,0 % de los casos y los fumadores empedernidos en 47,0 %. Conclusiones: La diabetes mellitus y la hipertensión arterial predominaron como enfermedades personales y familiares asociadas, lo que muestra la estrecha relación entre estas y el síndrome metabólico. Deben considerarse la educación y promoción de salud en pacientes con síndrome metabólico desde la atención primaria, así como la verticalización de la atención médica para este grupo en instituciones hospitalarias, a fin de prevenir y controlar los factores de riesgo relacionados con su aparición.

Computational Evaluation and In Vitro Validation of New Epidermal Growth Factor Receptor Inhibitors.

BACKGROUND: The Epidermal Growth Factor Receptor (EGFR) is a transmembrane protein that acts as a receptor of extracellular protein ligands of the epidermal growth factor (EGF/ErbB) family. It has been shown that EGFR is overexpressed by many tumours and correlates with poor prognosis. Therefore, EGFR can be considered as a very interesting therapeutic target for the treatment of a large variety of cancers such as lung, ovarian, endometrial, gastric, bladder and breast cancers, cervical adenocarcinoma, malignant melanoma and glioblastoma. METHODS: We have followed a structure-based virtual screening (SBVS) procedure with a library composed of several commercial collections of chemicals (615,462 compounds in total) and the 3D structure of EGFR obtained from the Protein Data Bank (PDB code: 1M17). The docking results from this campaign were then ranked according to the theoretical binding affinity of these molecules to EGFR, and compared with the binding affinity of erlotinib, a well-known EGFR inhibitor. A total of 23 top-rated commercial compounds displaying potential binding affinities similar or even better than erlotinib were selected for experimental evaluation. In vitro assays in different cell lines were performed. A preliminary test was carried out with a simple and standard quick cell proliferation assay kit, and six compounds showed significant activity when compared to positive control. Then, viability and cell proliferation of these compounds were further tested using a protocol based on propidium iodide (PI) and flow cytometry in HCT116, Caco-2 and H358 cell lines. RESULTS: The whole six compounds displayed good effects when compared with erlotinib at 30 µM. When reducing the concentration to 10µM, the activity of the 6 compounds depends on the cell line used: the six compounds showed inhibitory activity with HCT116, two compounds showed inhibition with Caco-2, and three compounds showed inhibitory effects with H358. At 2 µM, one compound showed inhibiting effects close to those from erlotinib. CONCLUSION: Therefore, these compounds could be considered as potential primary hits, acting as promising starting points to expand the therapeutic options against a wide range of cancers.

La COVID-19 en personas hipertensas / COVID-19 in hypertensive people

La COVID-19 es una enfermedad infecciosa causada por el coronavirus SARS-CoV-2, que afecta de forma más grave a personas en edades avanzadas de la vida y a pacientes con inmunodepresión y/o afecciones crónicas, como la hipertensión arterial, de gran significación en la mortalidad por enfermedades cardiovasculares y cerebrovasculares. Con este artículo se buscó actualizar los conocimientos sobre el nivel de gravedad de la COVID-19 en pacientes hipertensos y su asociación con el consumo de fármacos antihipertensivos de los grupos de los inhibidores de la enzima convertidora de la angiotensina y los antagonistas de los receptores de la angiotensina II. Asimismo, se ofrecen evidencias científicas acerca de que la hipertensión arterial es un predictor clínico de gravedad en pacientes con dicha enfermedad infecciosa, lo cual se manifiesta sobre todo en las edades mayores de 60 años, y de que la suspensión de forma preventiva de los mencionados tratamientos antihipertensivos puede conducir a inestabilidad clínica y a resultados desfavorables.

COVID-19 is an infectious disease caused by the SARS-CoV-2 coronavirus that affects people in advanced ages of life, patients with immunodepression and/or chronic affections, as hypertension, in a more serious way, being the latter of great significance in the mortality due to cardiovascular and cerebrovascular diseases. With this work we wanted to update the knowledge on the COVID-19 serious condition level in hypertensive patients and its association with the consumption of antihypertensive drugs of the angiotensin converting enzyme inhibitors and antagonists of angiotensin II receptors groups. In the same way, scientific evidences are offered on the fact that hypertension is a clinical predictor of a serious condition in patients with this infectious disease, that is manifested mainly in people older than 60 years, and that the suspension in a preventive way of the mentioned antihypertensive treatments can lead to clinical instability and unfavorable results.

Aportes de las investigaciones pedagógicas a la educación médica cubana / Contribution of the pedagogicalal investigations to the Cuban medical education

Introducción: Los retos de la educación superior en el área de las ciencias médicas precisan de un nuevo enfoque, fundamentado en los principios de excelencia, calidad y pertinencia; de ahí la importancia de las investigaciones pedagógicas para el perfeccionamiento continuo del proceso docente-educativo en la educación médica. Objetivos: Determinar los principales aportes de las investigaciones pedagógicas en la educación médica cubana. Métodos: Se realizó una investigación descriptiva, de corte pedagógico, de 80 tesis, obtenidas del repositorio de tesis doctorales del sitio Infomed, en el período de enero a diciembre del 2017. Resultados: En la investigación predominaron las tesis de obtención del grado científico de Doctor en Ciencias Pedagógicas, con 65 de ellas, y 50,0 % correspondieron a la didáctica de la educación superior. Igualmente, se destacaron los aportes teóricos y prácticos que desarrollan potencialidades metodológicas y, a la vez, contribuyen a la formación lógica de los profesionales, con lo cual se reveló su esencia pedagógica. Conclusiones: Se proyectó una visión de futuro en la formación de los profesionales, con las potencialidades necesarias para su desempeño y un carácter humanista, así como el marco ético pertinente para satisfacer las necesidades de la salud en Cuba y en otros pueblos.

Introduction: The challenges of higher education in the area of the medical sciences need a new approach, based on the principles of excellence, quality and pertinence; so the importance of the pedagogical investigations for the continuous improvement of the teaching-educational process in the medical education. Objectives: To determine the main contributions of the pedagogical investigations in the Cuban medical education. Methods: A descriptive and pedagogical investigation of 80 theses, obtained from the Infomed doctoral thesis repository, was carried out from January to December, 2017. Results: In the study, theses to obtain the scientific degree of Doctor in Pedagogical Sciences prevailed, with 65 of them, and 50.0% of the dissertations corresponded to the didactics of higher education. Likewise, the theoretical and practical contributions that develop methodological potentialities were remarkable and, at the same time, they contribute to the logical training of professionals, with which their pedagogical essence was revealed. Conclusions: A future vision in training the professionals was projected, with the necessary potentialities for their performance, a humanist character and the pertinent ethical framework to satisfy health necessities in Cuba and in other countries.

ZNF518B gene up-regulation promotes dissemination of tumour cells and is governed by epigenetic mechanisms in colorectal cancer.

Most of colorectal cancer CRC-related death is due to metastasis and the finding of markers for prognosis of invasiveness, constitutes an appealing challenge. Here, after analysing cDNA array containing 43 tumour and 5 normal mucosa samples, we report that the expression of the ZNF518B gene as a whole and that of its two major splicing isoforms are significantly increased in tumours. The canonical isoform was also up-regulated in a patients' cohort containing 70 tumour and 69 adjacent tissue samples. The effects of silencing ZNF518B on the phenotype of CRC cell lines were then studied. The gene does not affect cell proliferation, but plays a significant role in cell migration and invasiveness and induces changes in the epithelial-to-mesenchymal transition markers, suggesting that ZNF518B favours tumour cell dissemination. To study the regulation of the gene, transcription-related changes in nucleosomal organisation and epigenetic marks around the transcriptional start site were analysed. The positioning of a nucleosome over the transcription start site and the differential presence of the epigenetic marks H3K9ac, H3K27ac, H3K4me3 and H3K9me3 correlate with gene expression. Inhibition of histone deacetylases increases the transcription of ZNF518B, which may be a candidate for invasiveness prognosis in CRC and a target for epigenetic drugs.

NRF2 through RPS6 Activation Is Related to Anti-HER2 Drug Resistance in HER2-Amplified Gastric Cancer.

PURPOSE: Despite the clinical advantage of the combination of trastuzumab and platinum-based chemotherapy in HER2-amplified tumors, resistance will eventually develop. The identification of molecular mechanisms related to primary and acquired resistance is needed. EXPERIMENTAL DESIGN: We generated lapatinib- and trastuzumab-resistant clones deriving from two different HER2-amplified gastric cancer cell lines. Molecular changes such as protein expression and gene-expression profile were evaluated to detect alterations that could be related to resistance. Functional studies in vitro were corroborated in vivo. The translational relevance of our findings was verified in a patient cohort. RESULTS: We found RPS6 activation and NRF2 to be related to anti-HER2 drug resistance. RPS6 or NRF2 inhibition with siRNA reduced viability and resistance to anti-HER2 drugs. In knockdown cells for RPS6, a decrease of NRF2 expression was demonstrated, suggesting a potential link between these two proteins. The use of a PI3K/TORC1/TORC2 inhibitor, tested in vitro and in vivo, inhibited pRPS6 and NRF2 expression and caused cell and tumor growth reduction, in anti-HER2-resistant models. In a cohort of HER2-amplified patients treated with trastuzumab and chemotherapy, a high level of NRF2 at baseline corresponds with worse progression-free survival. CONCLUSIONS: NRF2 through the PI3K/AKT/mTOR/RPS6 pathway could be a potential effector of resistance to anti-HER2 drugs in our models. RPS6 inhibition decreases NRF2 expression and restores sensitivity in HER2-amplified gastric cancer in vitro and in vivo. High NRF2 expression in gastric cancer patients predicts resistance to treatment. RPS6 and NRF2 inhibition could prevent resistance to anti-HER2 drugs.

Rigidez arterial como marcador de daño vascular en pacientes con hipertensión arterial controlada / Arterial stiffness as marker of vascular damage in patients with controlled arterial hypertension

Se realizó un estudio observacional, descriptivo y transversal de 27 adultos con hipertensión arterial controlada, seleccionados aleatoriamente en el Consultorio del Médico de la Familia No. 6 del Policlínico Docente Armando García Aspurú en Santiago de Cuba, durante el periodo de septiembre de 2015 a igual mes de 2016, con vistas a determinar la alteración de la rigidez arterial en ellos. En la serie se obtuvo un predominio del sexo femenino (69,2 por ciento) en relación con el masculino (30,7 por ciento) y la edad media fue de 48,96 años; igualmente, se identificó un elevado porcentaje con obesidad premórbida (48,1), a la cual le siguió el sobrepeso, con 33,3 por ciento), y el peso normal, con 18,3 por ciento). Se encontró 19,0 por ciento) con índice de rigidez grave; condición más sensible y alarmante que el índice tobillo-brazo como marcador de daño vascular.

An observational, descriptive and cross-sectional study of 27 adults with controlled hypertension, ramdomly selected in the family's physician No. 6 of Armando García Aspurú Teaching Polyclinic in Santiago de Cuba was carried out during the period of September, 2015 to same month in 2016, aimed at determining the change of arterial stiffness in them. In the series a prevalence of the female sex was obtained (69.2 percent) in connection with the male sex (30.7 percent) and the mean age was of 48.96 years; equally, a high percentage was identified with premorbid obesity (48.1), followed by overweight, with 33.3 percent, and normal weight, 18.3 percent. A 19.0 percent with severe stiffness index was found; a more sensitive and alarming condition than the index ankle-arm as marker of vascular damage.