Evolution Meets Disease: Penetrance and Functional Epistasis of Mitochondrial tRNA Mutations

About half of the mitochondrial DNA (mtDNA) mutations causing diseases in humans occur in tRNA genes. Particularly intriguing are those pathogenic tRNA mutations than can reach homoplasmy and yet show very different penetrance among patients. These mutations are scarce and, in addition to their obvious interest for understanding human pathology, they can be excellent experimental examples to model evolution and fixation of mitochondrial tRNA mutations. To date, the only source of this type of mutations is human patients. We report here the generation and characterization of the first mitochondrial tRNA pathological mutation in mouse cells, an m.3739G>A transition in the mitochondrial mt-Ti gene. This mutation recapitulates the molecular hallmarks of a disease-causing mutation described in humans, an m.4290T>C transition affecting also the human mt-Ti gene. We could determine that the pathogenic molecular mechanism, induced by both the mouse and the human mutations, is a high frequency of abnormal folding of the tRNAIle that cannot be charged with isoleucine. We demonstrate that the cells harboring the mouse or human mutant tRNA have exacerbated mitochondrial biogenesis triggered by an increase in mitochondrial ROS production as a compensatory response. We propose that both the nature of the pathogenic mechanism combined with the existence of a compensatory mechanism can explain the penetrance pattern of this mutation. This particular behavior can allow a scenario for the evolution of mitochondrial tRNAs in which the fixation of two alleles that are individually deleterious can proceed in two steps and not require the simultaneous mutation of both

Circulating Tumor Cells in Hepatocellular Carcinoma: A Comprehensive Review and Critical Appraisal

Hepatocellular carcinoma (HCC) is the fifth most common neoplasm and a major cause of cancer-related death worldwide. There is no ideal biomarker allowing early diagnosis of HCC and tumor surveillance in patients receiving therapy. Liquid biopsy, and particularly circulating tumor cells (CTCs), have emerged as a useful tool for diagnosis and monitoring therapeutic responses in different tumors. In the present manuscript, we evaluate the current evidence supporting the quantitative and qualitative assessment of CTCs as potential biomarkers of HCC, as well as technical aspects related to isolation, identification, and classification of CTCs. Although the dynamic assessment of CTCs in patients with HCC may aid the decision-making process, there are still many uncertainties and technical caveats to be solved before this methodology has a true impact on clinical practice guidelines. More studies are needed to identify the optimal combination of surface markers, to increase the efficiency of ex-vivo expansion of CTCs, or even to target CTCs as a potential therapeutic strategy to prevent HCC recurrence after surgery or to hamper tumor progression and extrahepatic spreading

Sensitivity to anti-Fas is independent of increased cathepsin D activity and adrenodoxin reductase expression occurring in NOS-3 overexpressing HepG2 cells

Stable overexpression of endothelial nitric oxide synthase (NOS-3) in HepG2 cells (4TO-NOS) leads to increased nitro-oxidative stress and upregulation of the cell death mediators p53 and Fas. Thus, NOS-3 overexpression has been suggested as a useful antiproliferative mechanism in hepatocarcinoma cells. We aimed to identify the underlying mechanism of cell death induced by NOS-3 overexpression at basal conditions and with anti-Fas treatment. The intracellular localization of NOS-3, the nitro-oxidative stress and the mitochondrial activity were analysed. In addition, the protein expression profile in 4TO-NOS was screened for differentially expressed proteins potentially involved in the induction of apoptosis. NOS-3 localization in the mitochondrial outer membrane was not associated with changes in the respiratory cellular capacity, but was related to the mitochondrial biogenesis increase and with a higher protein expression of mitochondrial complex IV. Nitro-oxidative stress and cell death in NOS-3 overexpressing cells occurred with the expression increase of pro-apoptotic genes and a higher expression/activity of the enzymes adrenodoxin reductase mitochondrial (AR) and cathepsin D (CatD). CatD overexpression in 4TO-NOS was related to the apoptosis induction independently of its catalytic activity. In addition, CatD activity inhibition by pepstatin A was not effective in blocking apoptosis induced by anti-Fas. In summary, NOS-3 overexpression resulted in an increased sensitivity to anti-Fas induced cell death, independently of AR expression and CatD activityThis study was supported by the Instituto de Salud Carlos III (FIS 09/00185). G. Ferrín was supported by the Networked Biomedical Research Center Hepatic and Digestive Diseases (CIBEREHD

Enumeration and Characterization of Circulating Tumor Cells in Patients with Hepatocellular Carcinoma Undergoing Transarterial Chemoembolization

Circulating tumor cells (CTCs), and particularly circulating cancer stem cells (cCSC), are prognostic biomarkers for different malignancies and may be detected using liquid biopsies. The ex vivo culture of cCSCs would provide valuable information regarding biological aggressiveness and would allow monitoring the adaptive changes acquired by the tumor in real time. In this prospective pilot study, we analyzed the presence of EpCAM+ CTCs using the IsoFlux system in the peripheral blood of 37 patients with hepatocellular carcinoma undergoing transarterial chemoembolization (TACE). The average patient age was 63.5 ± 7.9 years and 91.9% of the patients were men. All patients had detectable CTCs at baseline and 20 patients (54.1%) showed CTC aggregates or clusters in their peripheral blood. The increased total tumor diameter (OR: 2.5 (95% CI: 1.3–4.8), p = 0.006) and the absence of clusters of CTCs at baseline (OR: 0.2 (95% CI: 0.0–1.0), p = 0.049) were independent predictors of a diminished response to TACE. Culture of cCSC was successful in five out of thirty-three patients, mostly using negative enrichment of CD45− cells, ultra-low adherence, high glucose, and a short period of hypoxia followed by normoxia. In conclusion, the identification of clusters of CTCs before TACE and the implementation of standardized approaches for cCSC culture could aid to predict outcomes and to define the optimal adjuvant therapeutic strategy for a true personalized medicine in hepatocellular carcinoma

Development of a prediction model for short-term remission of patients with Crohn’s disease treated with anti-TNF drugs

Therapy with anti-tumor necrosis factor (TNF) has dramatically changed the natural history of Crohn’s disease (CD). However, these drugs are not without adverse events, and up to 40% of patients could lose efficacy in the long term. We aimed to identify reliable markers of response to anti-TNF drugs in patients with CD. A consecutive cohort of 113 anti-TNF naive patients with CD was stratified according to clinical response as short-term remission (STR) or non-STR (NSTR) at 12 weeks of treatment. We compared the protein expression profiles of plasma samples in a subset of patients from both groups prior to anti-TNF therapy by SWATH proteomics. We identified 18 differentially expressed proteins (p ≤ 0.01, fold change ≥ 2.4) involved in the organization of the cytoskeleton and cell junction, hemostasis/platelet function, carbohydrate metabolism, and immune response as candidate biomarkers of STR. Among them, vinculin was one of the most deregulated proteins (p < 0.001), whose differential expression was confirmed by ELISA (p = 0.054). In the multivariate analysis, plasma vinculin levels along with basal CD Activity Index, corticosteroids induction, and bowel resection were factors predicting NSTR

CREACIÓN E IMPLEMENTACIÓN DE UN SISTEMA DE SEGURIDAD Y SALUD OCUPACIONAL EN LA EMPRESA "INDALIST S.A." DEL CANTÓN EL CARMEN, 2013.

EL PRESENTE PROYECTO CONSISTE EN LA IMPLEMENTACIÓN DE UN SISTEMA DE SEGURIDAD Y SALUD OCUPACIONAL EN INDUSTRIAS INDALIST S.A, NO CONTABA CON UN SISTEMA DE SEGURIDAD Y SALUD OCUPACIONAL Y DESARROLLABA SUS ACTIVIDADES SIN EL CONTROL ADECUADO PARA SALVAGUARDAR LA VIDA DE LOS TRABAJADORES, PRESENTÁNDOSE FACTORES DE RIESGO FÍSICOS, MECÁNICOS, QUÍMICO, ETC., EN LAS DISTINTAS ÁREAS DE TRABAJO.THIS PROJECT INVOLVES THE IMPLEMENTATION OF A SYSTEM OF OCCUPATIONAL SAFETY AND HEALTH INDUSTRIES INDALIST SA, WHICH DID NOT HAVE A SAFETY AND OCCUPATIONAL HEALTH AND DEVELOPED ITS ACTIVITIES WITHOUT ADEQUATE CONTROL TO SAFEGUARD THE LIVES OF WORKERS, INTRODUCING FACTORS PHYSICAL RISK, MECHANICAL, CHEMICAL, ETC., IN DIFFERENT AREAS OF WORK

Activation of mTOR Signaling Pathway in Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) is the most frequent primary liver cancer and occurs mainly in patients with liver cirrhosis. The mammalian target of rapamycin (mTOR) signaling pathway is involved in many hallmarks of cancer including cell growth, metabolism re-programming, proliferation and inhibition of apoptosis. The mTOR pathway is upregulated in HCC tissue samples as compared with the surrounding liver cirrhotic tissue. In addition, the activation of mTOR is more intense in the tumor edge, thus reinforcing its role in HCC proliferation and spreading. The inhibition of the mTOR pathway by currently available pharmacological compounds (i.e., sirolimus or everolimus) is able to hamper tumor progression both in vitro and in animal models. The use of mTOR inhibitors alone or in combination with other therapies is a very attractive approach, which has been extensively investigated in humans. However, results are contradictory and there is no solid evidence suggesting a true benefit in clinical practice. As a result, neither sirolimus nor everolimus are currently approved to treat HCC or to prevent tumor recurrence after curative surgery. In the present comprehensive review, we analyzed the most recent scientific evidence while providing some insights to understand the gap between experimental and clinical studies.This work was supported by the research project PI18/01736 integrated into the Plan Estatal de I+D+I 2013-2016 and co-financed by the ISCIII-Subdirección General de Evaluación y Fomento de la Investigación and the Fondo Europeo de Desarrollo Regional (FEDER).Ye

ANÁLISIS DE LOS PROTOCOLOS DE ATENCIÓN SOBRE VIOLENCIA DE GÉNERO Y SU PERSPECTIVA EN EL ECUADOR

Situations of gender violence are considered a social problem that affects the world, under power relations, where one wants to dominate the other. This demands current research and creation of policies and protocols to guide the performance of service operators. The research was carried out using a qualitative methodology, descriptive scope, through content analysis and text interpretation; through the following units of analysis: protocol approach, theoretical paradigm from psychology, declaration of psychotherapeutic care model, care procedures for the victim and the aggressor, instruments of psychological assessment. The general objective was to analyze the attention protocols on gender violence. Out of a total of twelve protocols, selected that include the European Union, Central and South America, theoretical approaches based on gender, rights, by life cycle, ecological, multidisciplinary model and social equity were evidenced; It is important to highlight that the protocol of action of the Ministry of Education of Ecuador considers the victim, aggressor and observer as part of the intervention. The study revealed that only one protocol establishes psychological and restorative care for the aggressors, the remaining ten protocols focus their attention on the victim. Keywords: Gender violence, protocols, psychological care, victims, aggressors. References [1]Organización Mundial de la Salud y Escuela de Higiene y Medicina Tropical de Londres, Consejo Sudafricano de Investigaciones Médicas, «Estimaciones mundiales y regionales de la violencia contra la mujer: prevalencia y efectos de la violencia conyugal y de la violencia sexual no conyugal en la salud,» Ginebra , 2013. [2]Comisión Económica para América Latina y el Caribe, «Oservatorio de igualdad de género de América Latina y el Caribe,» [Online]. Available: https://oig.cepal.org/es/indicadores/feminicidio. [Last access: 28 02 2020]. [3]Comisión Ecuménica de Derechos Humanos, «CEDHU,» 21 11 2019. [Online]. Available: http://www.cedhu.org/noticias/boletines/221-femicidios-en-ecuador-la-violencia-contra-las-mujeres-no-para. [Last access: 28 02 2020]. [4]Coalición Nacional de Mujeres del Ecuador, «Informe sobra de seguimiento a la implementación de las recomendaciones del comité de la CEDAW al Estado Ecuatoriano» Quito, 2017. [5]Ministerio de Salud Pública , «Norma técnica para la elaboración de instrumentos normativos del Ministerio de Salud Pública», Quito , 2013. [6]C. Sanz and A. Sánchez. «Protocolizar las actividades de Enfermía,» Rol de Enfermería, vol. 24, nº 1, pp. 67-74, 2001. [7]Secretaria Nacional de Planificación y Desarrollo de Ecuador, «Agenda zonal zona 8 - Guayaquil», Quito, 2015. [8]Organización Mundial de la salud , «WHO Global Consultation on Violence and a public health priority,» Documento inédito, Ginebra, 1996. [9]Organización Panamericana de Salud y Organización Mundial de Salud, «Informe mundial sobre: la violencia y la salud: resumen,» Washington, D.C., 2002. [10]I. Romero, «Intervención en violencia de género.Consideraciones en torno al tratamiento.,» Intervención psicosocial, vol. 2, nº 19, pp. 191-199, 2010. &nbsp;Las situaciones de violencia de género se consideran como un problema social que afecta a nivel mundial, bajo relaciones de poder, donde uno quiere dominar al otro. Lo cual demanda actuales investigaciones y creaciones de políticas y protocolos que guíen la actuación de los operadores de servicios. La investigación se realizó, mediante una metodología cualitativa, alcance descriptivo, mediante el análisis de contenido e interpretación de textos; mediante las siguientes unidades de análisis: enfoque del protocolo, paradigma teórico desde la psicología, declaratoria de modelo de atención psicoterapéutico, procedimientos de atención para la víctima y al agresor, instrumentos de valoración psicológica. El objetivo general fue analizar los protocolos de atención sobre violencia de género. De un total de doce protocolos, seleccionados que incluyen la Unión Europea, Centro y sur américa, se evidenciaron como resultado enfoques teóricos basados en género, derechos, por ciclo de vida, modelo ecológico, multidisciplinar y equidad social; es importante destacar que el protocolo de actuación del Ministerio de Educación del Ecuador, considera a la víctima, agresor y observador, como parte de la intervención. El estudio revelo que solo un protocolo establece atención psicológica y restaurativa para los agresores, los diez protocolos restantes centralizan su atención en la víctima. Palabras Clave: Violencia de género, protocolos, atención psicológica, victimas, agresores. Referencias [1]Organización Mundial de la Salud y Escuela de Higiene y Medicina Tropical de Londres, Consejo Sudafricano de Investigaciones Médicas, «Estimaciones mundiales y regionales de la violencia contra la mujer: prevalencia y efectos de la violencia conyugal y de la violencia sexual no conyugal en la salud,» Ginebra , 2013. [2]Comisión Económica para América Latina y el Caribe, «Oservatorio de igualdad de género de América Latina y el Caribe,» [En línea]. Disponible en: https://oig.cepal.org/es/indicadores/feminicidio. [Último acceso: 28 02 2020]. [3]Comisión Ecuménica de Derechos Humanos, «CEDHU,» 21 11 2019. [En línea]. Disponible en: http://www.cedhu.org/noticias/boletines/221-femicidios-en-ecuador-la-violencia-contra-las-mujeres-no-para. [Último acceso: 28 02 2020]. [4]Coalición Nacional de Mujeres del Ecuador, «Informe sobra de seguimiento a la implementación de las recomendaciones del comité de la CEDAW al Estado Ecuatoriano» Quito, 2017. [5]Ministerio de Salud Pública , «Norma técnica para la elaboración de instrumentos normativos del Ministerio de Salud Pública», Quito , 2013. [6]C. Sanz y A. Sánchez. «Protocolizar las actividades de Enfermía,» Rol de Enfermería, vol. 24, nº 1, pp. 67-74, 2001. [7]Secretaria Nacional de Planificación y Desarrollo de Ecuador, «Agenda zonal zona 8 - Guayaquil», Quito, 2015. [8]Organización Mundial de la salud , «WHO Global Consultation on Violence and a public health priority,» Documento inédito, Ginebra, 1996. [9]Organización Panamericana de Salud y Organización Mundial de Salud, «Informe mundial sobre: la violencia y la salud: resumen,» Washington, D.C., 2002. [10]I. Romero, «Intervención en violencia de género.Consideraciones en torno al tratamiento.,» Intervención psicosocial, vol. 2, nº 19, pp. 191-199, 2010

Isolation of mitochondria for biogenetical studies: An update

The use of good quality preparations of isolated mitochondria is necessary when studying the mitochondrial biogenetical activities. This article explains a fast and simple method for the purification of mammalian mitochondria from different tissues and cultured cells, that is suitable for the analysis of many aspects of the organelle's biogenesis. The mitochondria isolated following the protocol described here, are highly active and capable of DNA. RNA and protein synthesis. Mitochondrial tRNA aminoacylation, mtDNA-protein interactions and specific import of added proteins into the organelles, can also be studied using this kind of preparations. (C) 2009 Elsevier B.V. and Mitochondria Research Society. All rights reserved